Anti-nampt antibodies and uses thereof

Abstract

To provide effective prophylactic and therapeutic agents for reducing morbidity and mortality of inflammatory disorders and of those observed in patients, particularly in critically ill patients, suffering from these disorders.SOLUTION: Provided are an anti-nicotinamide phosphoribosyltransferase (NAMPT) antibody or an antigen-binding fragment thereof, and a method for treating a subject having a NAMPT-associated local and / or systemic inflammatory disorder, the method comprising administering to the subject an effective amount of the antibody or the antigen-binding fragment.SELECTED DRAWING: None

Description

[Technical Field]

[0001] Federally sponsored research or development This invention was made with funding from the National Institutes of Health (NIH) under grant number R41 HL110707 Produced with U.S. Government support under STTR and R42 HL152888. The government has certain rights in this invention.

[0002] Related Applications This application claims priority from U.S. Provisional Application No. 62 / 883,952, filed August 7, 2019. The entire contents of the aforementioned priority application are incorporated herein by reference.

[0003] Sequence Listing This application has been submitted electronically in ASCII format and is incorporated herein by reference in its entirety. The above ASCII copy, created on July 1, 2020, contains the sequence listing A105818_1020WO_SL.txt, and its size is 53767 bytes. [Background technology]

[0004] The nicotinamide phosphoribosyltransferase (NAMPT) gene encodes the Catalyzes the condensation of amides with 5-phosphoribosyl-1-diphosphate to give nicotinamide mononucleotides. This protein encodes a protein that produces nicotinic acid phosphoribosyltransferase. It belongs to the silyltransferase (NAPRTase) family and is involved in metabolism, stress It is thought to be involved in many important biological processes, including immune response and aging.

[0005] Nicotinamide phosphoribosyltransferase (NAMPT) is a cytosolic NAMP It exists as both the T protein and the extracellular NAMPT (eNAMPT) protein. eNAMPT is secreted into the blood and mediates NF-κB signaling via ligation of Toll-like receptor 4 (TLR4). It functions as a cytokine / enzyme (cytozyme) that activates κB signaling, resulting in rapid It may further serve as a biomarker for inflammatory lung disorders such as chronic respiratory distress syndrome. Summary of the Invention [Problem to be solved by the invention]

[0006] According to the present specification, anti-NAMPT antibodies, including humanized antibodies, useful for therapeutic purposes are provided. Current clinical treatments for acute and chronic inflammatory lung disorders (e.g., ARDS, VILI) Known treatments are merely supportive. Therefore, the art does not provide any information on the treatment of inflammatory disorders. and the morbidity and mortality observed in patients suffering from these disorders, especially in severely ill patients. There is an unmet need for effective prophylactic and therapeutic agents to reduce [Means for solving the problem]

[0007] In a first aspect, a human nicotinamide phosphoribosyltransferase according to the present invention is provided. ANTIBODY OR ANTIGEN-BINDING FRAGMENT THEREOF SPECIFICALLY BINDING TO NAMPT ENZYME (NAMPT) - Patent application (i) a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 3, SEQ ID NO: 4 or 29 and the CDR2 domain having the amino acid sequence shown in SEQ ID NO: 5. (ii) a heavy chain variable region comprising a CDR3 domain having the amino acid sequence of SEQ ID NO: 6 or 1; CDR1 domain having the amino acid sequence shown in SEQ ID NO: 7, 12, 14, 33; 35 or 37, and a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 8. and a light chain variable region comprising a CDR3 domain having an amino acid sequence as set forth in claim 1.

[0008] In some embodiments of the above aspects, the isolated antibody or its antigen-binding fragment The heavy chain variable region of the present invention comprises: (a) a CDR having the amino acid sequence shown in SEQ ID NO: 3; 1 domain, a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 4, and a CDR3 domain having the amino acid sequence shown in SEQ ID NO: (b) a CDR3 domain having the amino acid sequence set forth in SEQ ID NO: 3; a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 29; a CDR2 domain having the amino acid sequence shown in SEQ ID NO:5; and a CDR3 domain having the amino acid sequence shown in SEQ ID NO:5. Includes:

[0009] In some embodiments of the above aspects, the isolated antibody or its antigen-binding fragment The light chain variable region of the present invention comprises: (a) a CDR having the amino acid sequence set forth in SEQ ID NO:6; 1 domain, a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 7, and a CDR3 domain having the amino acid sequence shown in SEQ ID NO: (b) a CDR3 domain having the amino acid sequence shown in SEQ ID NO: 6; a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 12; a CDR2 domain, and a CDR3 domain having the amino acid sequence shown in SEQ ID NO: 8; c) a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 6, a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 14 and a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 8. (d) a CDR3 domain having the amino acid sequence shown in SEQ ID NO: 6; a main CDR2 domain having the amino acid sequence shown in SEQ ID NO: 33, and a CDR2 domain having the amino acid sequence shown in SEQ ID NO: (e) a CDR3 domain having the amino acid sequence shown in SEQ ID NO: 6; a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 35; an R2 domain, and a CDR3 domain having the amino acid sequence shown in SEQ ID NO: 8; ) a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 6, a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 37, and a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 8. (g) a CDR3 domain having the amino acid sequence shown in SEQ ID NO: 11; a main CDR2 domain having the amino acid sequence shown in SEQ ID NO: 7, and a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 8 (h) a CDR3 domain having the amino acid sequence shown in SEQ ID NO: 11; a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 12; an R2 domain, and a CDR3 domain having the amino acid sequence shown in SEQ ID NO: 8; ) a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 11, a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 14, and a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 8. (j) a CDR1 domain having the amino acid sequence set forth in SEQ ID NO: 11; domain, a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 33, and SEQ ID NO: (k) a CDR3 domain having the amino acid sequence shown in SEQ ID NO: 11; CDR1 domain having the amino acid sequence set forth in SEQ ID NO: 35 a CDR2 domain and a CDR3 domain having the amino acid sequence set forth in SEQ ID NO: 8; or (l) a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 11, SEQ ID NO: 3 7, and a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 8. It contains a CDR3 domain having a nucleotide sequence.

[0010] In another embodiment, an isolated antibody according to the present invention that specifically binds to human NAMPT. or an antigen-binding fragment thereof, comprising: (a) a C having the amino acid sequence set forth in SEQ ID NO:3 a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 4, a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 5, and a CDR3 domain having the sequence shown in SEQ ID NO: 6. a heavy chain variable region comprising a CDR3 domain having the amino acid sequence shown in column number 5; and a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 6, an amino acid sequence shown in SEQ ID NO: 7, a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 8; (b) a light chain variable region comprising an R3 domain, and (c) a C having the amino acid sequence set forth in SEQ ID NO: 3. a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 4, a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 5, and a CDR3 domain having the sequence shown in SEQ ID NO: 6. a heavy chain variable region comprising a CDR3 domain having the amino acid sequence shown in column number 5; and a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 11; a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 8; (c) a light chain variable region comprising a CDR3 domain, and (c) an amino acid sequence as set forth in SEQ ID NO: 3. a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 4, and a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 5. a heavy chain variable region comprising a CDR3 domain having the amino acid sequence set forth in SEQ ID NO: 5; a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 11, a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 14, and a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 8. (d) a light chain variable region comprising a CDR3 domain having the amino acid sequence shown in SEQ ID NO: 3; a CDR1 domain having the amino acid sequence shown in SEQ ID NO:29; and a CDR2 domain having the amino acid sequence shown in SEQ ID NO:29. and a heavy chain variable region comprising a CDR3 domain having the amino acid sequence shown in SEQ ID NO:5. a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 11; a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 14; and a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 8. (e) a light chain variable region comprising a CDR3 domain having the amino acid sequence set forth in SEQ ID NO: 3; a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 29; a heavy chain comprising a CDR3 domain having the amino acid sequence set forth in SEQ ID NO:5; a variable region and a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 11; 33, and a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 8. (f) a light chain variable region comprising a CDR3 domain having the amino acid sequence shown in SEQ ID NO: 3; a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 29; a CDR2 domain and a CDR3 domain having the amino acid sequence shown in SEQ ID NO:5. a heavy chain variable region comprising a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 11; a CDR2 domain having the amino acid sequence shown in sequence number 35, and a CDR2 domain having the amino acid sequence shown in sequence number 8; (g) a light chain variable region comprising a CDR3 domain having an amino acid sequence shown in SEQ ID NO: 3; a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 29; a CDR2 domain having the amino acid sequence shown in SEQ ID NO:5; and a CDR3 domain having the amino acid sequence shown in SEQ ID NO:5. a heavy chain variable region comprising the amino acid sequence shown in SEQ ID NO: 11; and a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 12. a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 37, and a CDR3 domain having the amino acid sequence shown in SEQ ID NO: 8 (h) a light chain variable region comprising a CDR3 domain having the amino acid sequence shown; a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 4; and a CDR2 domain having the amino acid sequence shown in SEQ ID NO:5. a heavy chain variable region comprising a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 11; a main CDR2 domain having the amino acid sequence shown in SEQ ID NO: 33, and a CDR2 domain having the amino acid sequence shown in SEQ ID NO: (i) a light chain variable region comprising a CDR3 domain having the amino acid sequence shown in SEQ ID NO: 8; No. 3, a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 4 a CDR2 domain having the sequence shown in SEQ ID NO: 5, and a CDR having the amino acid sequence shown in SEQ ID NO: 6 A heavy chain variable region comprising three domains and a CDR having the amino acid sequence shown in SEQ ID NO: 11. 1 domain, a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 35, and the sequence a light chain variable region comprising a CDR3 domain having the amino acid sequence shown in number 8, or ) a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 3, a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 4 a CDR2 domain having the amino acid sequence shown in SEQ ID NO:5; and a heavy chain variable region comprising a CDR3 domain having the amino acid sequence shown in SEQ ID NO: 11. a CDR1 domain having the amino acid sequence set forth in SEQ ID NO: 37; and a light chain variable region comprising a CDR3 domain having the amino acid sequence set forth in SEQ ID NO: 8; Includes:

[0011] In some embodiments of the above aspects, the isolated antibody or its antigen-binding fragment The ment is humanized.

[0012] In some embodiments of any of the foregoing aspects, the isolated antibody or its The original binding fragment is shown in SEQ ID NO: 2, 10, 13, 30, 31, 32, 34 or 36. In some embodiments, the single The isolated antibody or antigen-binding fragment thereof is selected from the group consisting of SEQ ID NOs: 1, 9, 15, 16, and 28. It contains a heavy chain variable region having the amino acid sequence shown.

[0013] In another aspect, the present invention provides an isolated antibody or a method for the treatment of NAMPT. an antigen-binding fragment of the variable region comprising the amino acid sequence set forth in SEQ ID NO: 15; and a light chain comprising a variable region comprising the amino acid sequence set forth in SEQ ID NO: 13. The present invention features an isolated antibody or antigen-binding fragment thereof.

[0014] In a different embodiment, an isolated antibody that specifically binds to human NAMPT according to the present invention is provided. The antibody or antigen-binding fragment thereof has a variable region comprising the amino acid sequence set forth in SEQ ID NO:26. a heavy chain comprising a variable region and a light chain comprising an amino acid sequence set forth in SEQ ID NO: 18; include.

[0015] In another aspect, the present invention provides an isolated antibody or an antigen-binding fragment thereof, wherein the heavy chain comprises a variable region as set forth in Table 17; and a light chain comprising a variable region as set forth in Table 17. The binding fragments are characterized.

[0016] In yet another aspect, the present invention provides an isolated antibody that specifically binds to human NAMPT. or an antigen-binding fragment thereof, comprising: a heavy chain CDR1, a CD4, a DH ... a heavy chain comprising CDR1, CDR2 and CDR3, and a light chain comprising CDR1, CDR2 and CDR3 as set forth in Table 17; and a light chain comprising a CDR3. do.

[0017] In a different embodiment, the isolated anti-NAMPT antibody of the present invention is antibody AL-30. It contains three heavy and light chain variable regions.

[0018] In another aspect, the present invention provides an isolated anti-NAMPT antibody, comprising antibody AL-31 The present invention features an isolated anti-NAMPT antibody comprising a heavy chain variable region and a light chain variable region of NAMPT.

[0019] In yet another aspect, the present invention provides an isolated humanized anti-NAMPT antibody, The humanized heavy chain variable region derived from mouse antibody AL-303 or AL-310 and the humanized heavy chain variable region derived from mouse antibody AL an isolated anti-NAMP antibody comprising a humanized light chain variable region derived from AL-303 or AL-310; T antibodies.

[0020] In a different embodiment, the present invention provides a method for the preparation of ... An isolated anti-NAMPT antibody, comprising: (a) a sequence encoding amino acid residues 17 to 44 of SEQ ID NO: 60; at least one amino acid of amino acid residues 117 to 127 of SEQ ID NO: 60; at least one amino acid, at least one amino acid residue between 162 and 170 of SEQ ID NO: 60 At least one amino acid residue of amino acid residues 242 to 261 of SEQ ID NO: 60 at least one amino acid selected from amino acid residues 262 to 273 of SEQ ID NO: 60; amino acid, at least one amino acid selected from amino acid residues 289 to 305 of SEQ ID NO: 60 , at least one amino acid selected from amino acid residues 332 to 342 of SEQ ID NO: 60; At least one amino acid among amino acid residues 374 to 389 of SEQ ID NO: 60, SEQ ID NO: 6 at least one amino acid of amino acid residues 418 to 425 of SEQ ID NO: 60; At least one amino acid of amino acid residues 453 to 466 and the amino acid of SEQ ID NO: 60 The amino acid residues on human NAMPT contain at least one amino acid between 408 and 416. or (b) at least one of amino acid residues 29 to 51 of SEQ ID NO: 60. amino acids, at least one amino acid selected from amino acid residues 61 to 72 of SEQ ID NO: 60; At least one amino acid among amino acid residues 156 to 170 of SEQ ID NO: 60, SEQ ID NO: At least one amino acid of amino acid residues 216 to 234 of SEQ ID NO: 60 at least one amino acid of amino acid residues 316 to 331 of SEQ ID NO: 60; at least one amino acid of amino acid residues 332 to 342 of SEQ ID NO: 60 at least one amino acid of residues 373 to 389, amino acid residue 41 of SEQ ID NO: 60 at least one amino acid of amino acid residues 7 to 431 of SEQ ID NO: 60, 69, and amino acid residues 470-47 of SEQ ID NO: 60 Any of the epitopes on human NAMPT containing at least one amino acid of 8 The present invention features an isolated anti-NAMPT antibody that binds to an epitope on human NAMPT, including do.

[0021] In some embodiments of any of the above aspects, the antibody or antigen-binding fragment thereof The ment contains an Fc domain.

[0022] In some embodiments of any of the preceding aspects, the isolated antibody or its The antigen-binding fragment is a monoclonal antibody.

[0023] In some embodiments of any of the foregoing aspects, the isolated antibody or its antibody In some embodiments, the isolated antibody-binding fragment is an IgG. In some embodiments, the antibody or antigen-binding fragment thereof is an IgG1. The isolated antibody or antigen-binding fragment thereof is an IgG4.

[0024] In some embodiments, the present invention provides the isolated Nucleic acids encoding either the antibody or antigen-binding fragment thereof, vectors, and / or host cells containing said nucleic acid or said vectors.

[0025] In certain embodiments, the present invention provides the isolated antibodies featured herein. A pharmaceutical composition comprising either the antibody or an antigen-binding fragment thereof and a pharmaceutically acceptable carrier. Offer your body.

[0026] In some embodiments, the present invention provides an effective amount of any of the compounds featured herein. By administering either the isolated antibody or antigen-binding fragment to a subject in need thereof. and methods for treating disorders associated with deleterious NAMPT activity in a subject in need thereof. nothing.

[0027] Also featured herein is a method of treating a subject having an inflammatory condition. and administering an effective amount of the isolated antibody or antigen-binding fragment described herein to In some embodiments, the inflammatory disease is characterized by the inability to differentiate from the inflammatory disease. is a common cause of pulmonary fibrosis (IPF), pulmonary hypertension, acute lung injury (ALI), and acute respiratory distress syndrome (ARES). RDS), ventilator-induced lung injury (VILI), ARDS / VILI-induced ALI, Trauma-induced acute lung injury (TIALI) and brain injury or radiation-induced lung injury (e.g., cancer treatment) Radiation-induced lung injury caused by radiation related to

[0028] In some embodiments, the present invention provides a method for treating prostate cancer (PCa) in a subject in need thereof. 1. A method of treating a cancer comprising administering an effective amount of an isolated antibody or its derivatives described herein. and administering to the subject an antigen-binding fragment of the antibody. In some embodiments, the subject has recurrent PCa. The subject is at risk of developing metastatic PCa. In some embodiments, the tumor is resistant to androgen deprivation therapy (ADT). The method further comprises administering ADT to the subject. [Brief explanation of the drawings]

[0029] [Figure 1] 1 is a Western blot assay showing the detection of human NAMPT and mouse NAMPT by mouse anti-NAMPT antibodies AL-303, AL-304, AL-305, AL-309, and AL-310. [Figure 2] 1 is a Western blot assay showing the ability of mouse anti-NAMPT antibodies AL-303, AL-304, AL-305, AL-309 and AL-310 mAbs to reduce hNAMPT-induced NFκB phosphorylation. [Figure 3] FIG. 1 is a schematic diagram of the experimental protocol for an in vivo study to evaluate the effects of mouse anti-NAMPT antibodies AL-303, AL-304, and AL-305 on NAMPT-induced mouse lung injury. [Figure 4] Figure 4A shows the effect of mouse anti-NAMPT antibodies AL-303, AL-304, and AL-305 on NAMPT-induced mouse lung injury. Figure 4A shows the effect of mouse anti-NAMPT antibodies on NAMPT-induced bronchoalveolar lavage (BAL) protein levels. Figure 4B shows the effect of mouse anti-NAMPT antibodies on NAMPT-induced BAL-expressing polymorphonuclear neutrophil (PMN) counts (n = 3-6 mice, *P < 0.05). [Figure 5] FIG. 1 is a schematic diagram of the experimental protocol for an in vivo study to evaluate the effect of the murine anti-NAMPT antibody AL-310 on LPS-induced mouse lung injury. [Figure 6] 1 is a graph showing the effect of mouse anti-NAMPT antibody AL-310 on LPS-induced BAL protein levels. [Figures 7A-7B] Figure 7A is a graphical representation of the effect of humanized 1076 and 1093 anti-hNAMPT antibodies on hNAMPT-induced NFκB phosphorylation. Figure 7B is a graphical representation of the effect of humanized 1076 and 1093 anti-hNAMPT antibodies on hNAMPT-induced reduction in endothelial cell (EC) barrier integrity. [Figures 8A-8D]Figure 8 shows the effects of humanized 1076 and 1093 anti-hNAMPT antibodies on mouse and rat models of lung injury. Figure 8A is a graphical representation of the effects of 1076 anti-hNAMPT antibodies V-1076, N-1076, K-1076, and P-1076 on BAL protein levels (left panel of Figure 8A) and BAL PMN counts (right panel of Figure 8A) in a mouse model of lung injury. Figure 8B is a graphical representation of the effects of 1093 anti-hNAMPT antibodies SS-1093, CC-1093, XX-1093, and UU-1093 on BAL protein levels (left panel of Figure 8B) and BAL PMN counts (right panel of Figure 8B) in a mouse model of lung injury. Figure 8C is a graphical representation of the effect of the 1076 anti-hNAMPT antibody P-1076 on BAL protein levels (left panel of Figure 8C) and BAL PMN counts (right panel of Figure 8C) in a rat model of lung injury. Figure 8D shows H&E stained photomicrographs showing the effect of the 1093 anti-hNAMPT antibody UU-1093 on cellular infiltration and edema in a mouse model of lung injury. [Figures 9A-9D] Immunohistochemical (IHC) staining of NAMPT in normal, minimally invasive, and highly invasive prostate cancer (PCa) is shown. Figure 9A is a photomicrograph showing very low NAMPT expression in normal prostate tissue. Figure 9B is a photomicrograph showing modest but significantly increased NAMPT expression in gland-confined prostate adenocarcinoma. Figure 9C is a photomicrograph showing strong NAMPT expression within tumor cells in three separate prostate adenocarcinomas with penetration into the smooth muscle capsule and invasion into periprostatic adipose tissue. Figure 9D is a graph showing cumulative analysis of NAMPT expression in 26 PCa patients with organ-confined disease (n = 12) and capsular-invasive disease (n = 14). [Figures 10A-10D]Results from analysis of mouse and human tissues after acute and subacute radiation exposure are shown. Figure 10A provides photomicrographs of H&E staining showing evidence of inflammation and damage in mouse lung tissue one week after radiation exposure (lower panel of Figure 10A) and before exposure to a single dose of thoracic radiation (upper panel of Figure 10A). Figure 10B provides photomicrographs of IHC staining showing NAMPT expression in mouse lung tissue one week after radiation exposure (lower panel of Figure 10B) and before exposure to radiation (upper panel of Figure 10B). Figure 10C is a high-magnification image of the photomicrograph showing NAMPT expression in alveolar macrophages (long arrows) and pneumocytes (short arrows). Figure 10D shows photomicrographs showing NAMPT expression in normal, post-operative human tonsillar epithelial tissue that was exposed to 8 Gy of radiation for 24 hours ("irradiated") (lower panel of Figure 10D) or not exposed to radiation ("non-irradiated") (upper panel of Figure 10D). [Figures 11A-11B]

[0049] Figure 11A shows results from an in vitro assay of human DU-145 PCa cell migration through human smooth muscle cells. Figure 11A is a graph showing the number of invaded DU-145 PCa cells in the absence ("medium only") or presence ("NAMPT") of NAMPT, with medium without PCa cells ("No PCa cells") serving as a negative control. Figure 11B provides photomicrographs of wells with PCa cells cultured in the absence ("medium only") or presence ("NAMPT") of NAMPT, with wells without PCa cells ("No PCa cells") serving as a negative control. [Figures 12A-12C]Figure 12 shows results from in vivo testing of 1076 humanized anti-hNAMPT antibodies (N-1076, K-1076, and P-1076) and 1093 humanized anti-hNAMPT antibodies (SS-1093, XX-1093, and UU-1093) on inflammation and injury in a mouse lung injury model. Figure 12A is a graph showing the effect of humanized anti-hNAMPT antibodies on lung injury scores in an LPS-induced "one-hit" lung injury model. Figure 12B is a graph showing the effect of humanized anti-hNAMPT antibodies on lung injury scores in an LPS / VILI-induced "two-hit" lung injury model. Figure 12C provides photomicrographs showing histological indices of the effect of humanized anti-hNAMPT antibody P-1076 on lung injury in an LPS-induced "one-hit" lung injury model (upper panel of Figure 12C) and an LPS / VILI-induced "two-hit" lung injury model (lower panel of Figure 12C). [Figures 13A-13C] Figure 13 shows results from an in vivo assay of PCa cell invasion through diaphragm smooth muscle. Figure 13A is a photomicrograph showing severe peritoneal dissemination with invasion through the smooth muscle layer of a SCID mouse 6 weeks after intraperitoneal (IP) injection of highly metastatic human PCa cells, PC3; a magnified image of the photomicrograph is provided in the bottom panel of Figure 13A. Figure 13B is a photomicrograph showing inhibition of PC3 cell invasion in PC3-injected mice that received the humanized anti-hNAMPT antibody P-1076; a magnified image of the photomicrograph is provided in the bottom panel of Figure 13B. Figure 13C is a graph showing the percentage of PC3 cells invading the diaphragm of mice treated with the anti-hNAMPT antibody P-1076 or vehicle alone. [Figures 14A-14E]The effects of NAMPT neutralizing antibodies on lung inflammation (assessed by H&E staining), the amount of BAL protein, and the number of BAL-expressing cells, as assessed in a mouse model of RILI, are shown. Figure 14A provides representative photomicrographs showing H&E staining in lung tissue from vehicle control (left panel of Figure 14A), non-irradiated control mice (left panel of the inset in Figure 14A), or irradiated RILI mice injected with a vehicle control (left panel of Figure 14A), an anti-NAMPT polyclonal antibody (pAb) (middle panel of Figure 14A), or an anti-NAMPT monoclonal antibody (mAb) (right panel of Figure 14A) after radiation exposure. Figure 14B is a graphical representation of H&E staining (% area) in lung tissue from vehicle control, non-irradiated control mice, or irradiated RILI mice injected with an anti-NAMPT pAb or anti-NAMPT mAb. Figure 14C is a graphical representation of BAL protein levels (μg / ml) in the lung tissue of non-irradiated control mice or irradiated RILI mice injected with a vehicle control, anti-NAMPT pAb, or anti-NAMPT mAb. Figure 14D is a graphical representation of the number of BAL-expressing cells in the lung tissue of non-irradiated control mice or irradiated RILI mice injected with a vehicle control, anti-NAMPT pAb, or anti-NAMPT mAb. Figure 14E is a graphical representation of the ALI severity score of non-irradiated control mice or irradiated RILI mice injected with a vehicle control, anti-NAMPT pAb, or anti-NAMPT mAb. * indicates p<0.05. [Figures 15A-15D]Figure 15A shows the detection of NAMPT expression with a 99mTc-labeled anti-NAMPT mAb probe in a non-irradiated control mouse (left panel of Figure 15A) or an irradiated (RILI) mouse (PBI) exposed to 8 Gy partial body irradiation (right panel of Figure 15A). Figure 15B shows the detection of NAMPT expression with a 99mTc-labeled anti-NAMPT mAb probe in a non-irradiated control mouse (top panel of Figure 15B) or an irradiated (RILI) mouse (bottom panel of Figure 15B). Figure 15C shows a graphical representation of the ratio of lung activity to tissue background from the left and right lungs of a non-irradiated control mouse or an irradiated (RILI) mouse. Figure 15D shows a graphical representation of the radioactivity (%ID / g) in the lung tissue of a non-irradiated control mouse or an irradiated (RILI) mouse. * indicates p<0.05. [Figures 16A-16C] Figure 16 shows the effects of humanized anti-NAMPT mAb on BAL cell counts, collagen deposition, and lung tissue smooth muscle actin (SMA) expression, as assessed in a mouse model of RILI 18 weeks after 20 Gy radiation exposure. Figure 16A is a graph showing the number of BAL-expressing cells in the lung tissue of irradiated RILI mice intraperitoneally injected with anti-NAMPT mAb or vehicle control. Figure 16B provides representative images from Western blot analysis showing SMA expression in lung tissue homogenates of irradiated RILI mice intraperitoneally injected with anti-NAMPT mAb or vehicle control. Figure 16C provides representative photomicrographs showing collagen deposition detected by trichrome staining in the lung tissue of irradiated RILI mice intraperitoneally injected with anti-NAMPT mAb or vehicle control. * indicates p<0.05. [Figures 17A-17C]Figure 17 shows the effects of humanized anti-NAMPT mAb on inflammatory cell infiltration, edema, and lung injury score, as assessed in a rat model of trauma (blast injury) / ventilation-induced lung injury (VILI). Figure 17A provides representative images and photomicrographs showing lungs or lung tissue sections from trauma / VILI-challenged rats injected with vehicle control. The left panel of Figure 17A provides representative images of lungs from trauma / VILI-challenged rats injected with vehicle control. The middle and right panels of Figure 17A provide representative photomicrographs showing inflammatory cell infiltration and edema, as assessed by H&E staining, in trauma / VILI-challenged rats injected with vehicle control. The inset in the right panel of Figure 17A provides representative photomicrographs showing H&E staining in lung tissue from a rat not challenged with trauma / VILI. Figure 17B provides representative images and photomicrographs showing lungs or lung tissue sections from trauma / VILI-challenged rats injected with anti-NAMPT mAb. The left panel of Figure 17B provides representative images of lungs from trauma / VILI-challenged rats injected with anti-NAMPT mAb. The middle and right panels of Figure 17B provide representative photomicrographs showing inflammatory cell infiltration and edema, as assessed by H&E staining, in trauma / VILI-challenged rats injected with anti-NAMPT mAb. Figure 17C is a graph showing lung injury scores in trauma / VILI-challenged rats injected with either anti-NAMPT mAb or vehicle control. [Figures 18A-18C]Figure 18 shows the effects of NAMPT neutralizing antibodies on inflammatory cell infiltration, edema, and lung injury scores, as assessed in a rat model of murine LPS / VILI lung injury. Figure 18A provides representative photomicrographs showing inflammatory cell infiltration and edema, as assessed by H&E staining, in LPS / VILI-challenged mice injected with a vehicle control. The inset in Figure 18A provides representative photomicrographs showing H&E staining in lung tissue from mice not challenged with LPS / VILI. Figure 18B provides representative photomicrographs showing inflammatory cell infiltration and edema, as assessed by H&E staining, in LPS / VILI-challenged mice injected with an anti-NAMPT mAb. Figure 18C is a graph showing ALI severity scores, as assessed in LPS / VILI-challenged mice injected with an anti-NAMPT mAb, an anti-NAMPT pAb, or a vehicle control (PBS). The graph in Figure 18C also shows the ALI severity scores of control mice not challenged with LPS / VILI. * indicates p<0.05. *** indicates p<0.001. [Figures 19A-19D]Figure 19A shows the detection of NAMPT expression by a 99mTc-labeled anti-NAMPT mAb probe. Figure 19A provides representative autoradiographic images showing the detection of NAMPT expression by a 99mTc-labeled anti-NAMPT mAb probe (PRONAMPTOR) (right panel of Figure 19A) or a radiolabeled IgG control Ab (left panel of Figure 19A) in mice exposed to 20 Gy of whole lung irradiation (WTLI). Figure 19B provides representative autoradiographic images showing the detection of NAMPT expression by a 99mTc-labeled anti-NAMPT mAb probe in LPS-challenged mice (right panel of Figure 19B) or unchallenged control mice (left panel of Figure 19B) 3 hours after LPS challenge. Figure 19C provides a representative autoradiographic image showing detection of NAMPT expression by a 99mTc-labeled anti-NAMPT mAb probe in the lungs of LPS-challenged mice (bottom panel of Figure 19C) or unchallenged control mice (top panel of Figure 18C) 3 hours after LPS challenge. Figure 19D is a graphical representation of the uptake of the radiolabeled anti-NAMPT mAb probe, as assessed by radioactivity (%ID / g), in the lung tissues of LPS-challenged or unchallenged control mice 3 and 18 hours after LPS challenge. * indicates p<0.05. [Figures 20A-20B] Figure 20 shows the effect of humanized anti-NAMPT mAb on right ventricular systolic pressure (RVSP) and pulmonary artery thickness as assessed in the rat monocrotaline (MCT) model of PAH. Figure 20A is a graphical representation of RVSP in MCT-challenged rats injected with either anti-NAMPT mAb or vehicle control (control MCT mice). Figure 20B provides representative photomicrographs showing pulmonary artery thickness as assessed by H&E staining in MCT-challenged rats injected with either anti-NAMPT mAb (right panel of Figure 20B) or vehicle control (left panel of Figure 20B). * indicates p<0.05. DETAILED DESCRIPTION OF THE INVENTION

[0030] The present invention can be embodied in many different forms. 1 is a non-limiting exemplary embodiment of the present invention illustrating the principles of the present invention. Any section headings provided are for organizational purposes only and do not serve to identify the subject matter described. For the purposes of this disclosure, all identifying sequence alleles are intended to be illustrative and not to be limiting. Accession numbers are from the NCBI Reference Sequence (REFSEQ) database unless otherwise noted. and / or the NCBI GenBank® archive sequence database. can.

[0031] Various aspects of the present invention relate to anti-NAMPT antibodies and antibody fragments thereof, pharmaceutical compositions, and and nucleic acids, recombinant expression vectors and hosts for producing such antibodies and fragments. Detect human NAMPT and measure human NAMPT activity (in vitro or in vivo) in primary cells. inhibits the pulmonary fibrosis (IPF), pulmonary hypertension, acute lung injury (ALI), and acute respiratory distress. syndrome (ARDS), ventilator-induced lung injury (VILI), ARDS / VILI-induced ALI, trauma-induced acute lung injury (TIALI) and brain injury, radiation-induced lung injury, and NAMPT-related diseases, including but not limited to cancer (e.g., prostate cancer (PCa)) Methods of using the antibodies described herein for treatment are also included in the invention.

[0032] definition In order that the present invention may be more readily understood, certain terms are first defined. Note that whenever a parameter's value or range of values ​​is listed, the column Values ​​and ranges intermediate to the recited values ​​are also intended to be part of this invention.

[0033] The terms "NAMPT" and "eNAMPT" are used interchangeably herein. The term particularly relates to non-secreted (e.g., intracellular NAMPT or NAMPT nucleic acid) forms. Unless otherwise stated, the secreted form of nicotinamide phosphoribosyltransferase is referred to. The amino acid sequence of secreted human NAMPT (also called human eNAMPT) is SEQ ID NO: No. 60 (NCBI gene reference numbers NC_000007.14 and See also protein reference number NP_005737.1).

[0034] MNPAAEAEFN ILLATDSYKV THYKQYPPNT SKVYSY FECR EKKTENSKLR KVKYEETVFY GLQYILNKYL KGKVVTKEKI QEAKDVY KEH FQDDVFNEKG WNYILEKYDG HLPIEIKAVP EGFVIPRGNV LFTVENT DPE CYWLTNWIET ILVQSWYPIT VATNSREQKK ILAKYLLETS GNLDGLE YKL HDFGYRGVSS QETAGIGASA HLVNFKGTDT VAGLALIKKY YGTKDPV PGY SVPAAEHSTI TAWGKDHEKD AFEHIVTQFS SVPVSVVSDS YDIYNAC EKI WGEDLRHLIV SRSTQAPLII RPDSGNPLDT VLKVLEILGK KFPVTEN SKG YKLLPPYLRV IQGDGVDINT LQEIVEGMKQ KMWSIENIAF GSGGGLL QKL TRDLLNCSFK CSYVVTNGLG INVFKDPVAD PNKRSKKGRL SLHRTPA GNF VTLEEGKGDL EEYGQDLLHT VFKNGKVTKS YSFDEIRKNA QLNIELE AAHH (SEQ ID NO: 60) NAMPT is also known as pre-B cell colony-enhancing factor (PBEF) or visfatin. do.

[0035] "NAMPT antibody" or "anti-NAMPT antibody" are used interchangeably herein The term refers specifically to the secreted form of NAMPT (also referred to herein as eNAMPT). An antibody that "binds" to the antigen of interest, i.e., NAMPT, is an antibody that The antibody binds to the antigen with sufficient affinity so that it is useful for targeting cells that express the antigen. In a preferred embodiment, the antibody is capable of binding to human NAMPT ( It specifically binds to extracellular human NAMPT (human eNAMPT), particularly extracellular human NAMPT (human hNAMPT). Examples of anti-eNAMPT antibodies are disclosed in the Examples and Sequence Listing provided below.

[0036] As used herein, the "biological activity of NAMPT" includes binding to TLR4. This refers to all of the unique biological properties of NAMPT, including but not limited to:

[0037] Terms such as "specifically bind" or "specifically bind to" refer to the binding of an antibody or its antigen. The fragment forms a complex with an antigen that is relatively stable under physiological conditions, such as NAMPT. Specific binding means forming a specific bond with at least about 1 x 10 -8 Equilibrium dissociation constant below M can be characterized by a number (e.g., K D The smaller the value, the stronger the bond. Methods for determining whether two molecules specifically bind are well known in the art, and include: Examples include equilibrium dialysis and surface plasmon resonance.

[0038] The term "antibody" broadly refers to immunoglobulin (Ig) molecules, generally consisting of four Polypeptide chains, two heavy (H) chains and two light (L) chains, or the essential target of an Ig molecule Any functional fragment, mutant, variant or derivative of those chains that retains binding function. Such mutant, variant or derivative antibody formats are These are known in the art, non-limiting embodiments of which are discussed below.

[0039] In a full-length antibody, each heavy chain comprises a heavy chain variable region (abbreviated herein as HCVR or VH). It consists of a heavy chain constant region and a heavy chain constant region.

[0040] Both the light and heavy chains are divided into regions of structural and functional homology. The terms " and "variable" are used functionally. In this regard, it will be understood that The variable domains of both the variable light (VL) and variable heavy (VH) chain portions are responsible for antigen recognition and specificity. Conversely, the constant domains of the light chain (CL) and heavy chain (CH1, CH2 or CH3) are determined. confers biological properties such as secretion, transplacental transfer, Fc receptor binding, and complement fixation. Therefore, the numbering of the constant region domains is based on the antigen binding site or amino terminus of the antibody. The N-terminal part is a variable region, and the C-terminal part is a constant region. The CH3 (or CH4 in the case of IgM) and CL domains are actually the heavy chain and Each comprises the carboxy terminus of a light chain.

[0041] The heavy chain constant region is composed of three domains: CH1, CH2, and CH3. A chain comprises a light chain variable region (abbreviated herein as LCVR or VL) and a light chain constant region The light chain constant region is composed of one domain, CL. The VH and VL regions are , complementarity-determining regions (CDRs), interspersed with more conserved regions called framework regions (FRs). Each VH and VL can be further subdivided into regions of hypervariability called CDRs. is composed of three CDRs and four FRs, and from the amino terminus to the carboxy terminus: , FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4 Immunoglobulin molecules can be of any class (e.g., IgG, IgE, IgM, IgD, IgE ... IgA and IgY) and isotype (e.g., IgG1, IgG2, IgG3, IgG4 , IgA1 and IgA2) or subclasses. The light chains can be kappa or lambda (κ, λ) are classified into two types.

[0042] The "complementarity determining regions" or "CDRs" present in the antigen-binding domain of an antibody are each Specifically designed to form binding domains when the antibody assumes a three-dimensional structure in an aqueous environment. "Framework" or "FW" regions are short, non-contiguous sequences of amino acids that map to the The remainder of the amino acids in the binding domain of an antibody, called The binding domain formed by the positioned CDRs binds to an endogenous CDR on the immunoreactive antigen. This complementary surface defines the surface of the antibody that is complementary to its cognate epitope. The amino acids constituting the CDR and framework regions each promote non-covalent binding to the target peptide. Since the term "acid" is defined in a variety of different ways, as explained below, it is possible for a person skilled in the art to These can be readily identified for any given heavy or light chain variable region. Suitable CDRs are provided herein. a, Martin, PyIgClassify or IMGT to define CDRs Exemplary CDR definitions are set forth in the CDR section of the accompanying drawings, which are incorporated herein by reference in their entirety. "Antibody Structure and Function: The e Basis for Engineering Therapeutics”, An tibodies, 8(55):1-80(2019).

[0043] For the amino acid positions of the heavy chain constant region discussed in this invention, the numbering is Edelman et al., 1969, Proc., 1969, which describes the amino acid sequence of protein Eu. First published in Natl.Acad.Sci.USA 63(1):78-85 The EU index is the first sequenced human Ig The Edelman EU index is reported to be G1. 1 (see above). Therefore, the term "Kabat" in the context of heavy chains is "Kabat EU Index" or "Kabat EU Index" or "EU Index" The term "EU numbering" or "EU numbering" is derived from Kabat et al., 1991 (see above). As described in Edelman et al.'s human IgG1 Eu antibody, This refers to the numbering system used for the amino acid sequence of the light chain constant region. Systems similarly described in Kabat et al. (see above). The amino acid sequence of a typical kappa light chain constant region is set forth immediately below. RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKV QWKVDNALQSGNSQESVTEQDSKDSTYSLSSSTLTLSKADY EKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 59) Similarly, an exemplary IgG1 heavy chain constant region amino acid sequence compatible with the present invention is shown immediately below: is described in. ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTV SWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQ TYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLG GPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFN WYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNG KEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPSRD ELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPP VLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHY TQKSLSLSPGK (SEQ ID NO: 58)

[0044] As used herein, an "antigen-binding portion" or "binding fragment" of an antibody " refers to an antibody that retains the ability to specifically bind to an antigen (e.g., hNAMPT). The antigen-binding function of an antibody can be maintained by fragments of a full-length antibody. Such antibody embodiments also have been shown to be feasible by combining two or more different and a bispecific, bifunctional, or multispecific format that specifically binds to different antigens. Examples of binding fragments encompassed by the term "antigen-binding portion" of an antibody include (i) a monovalent fragment consisting of a VL domain, a VH domain, a CL domain, and a CH1 domain; (ii) a Fab fragment, which is a fragment of the Fab fragment, and (iii) a Fab fragment, which is a fragment of the Fab fragment, which is a fragment of the Fab fragment, which is a fragment of the Fab fragment, and (iv) a Fab fragment, which is a fragment of the ... and F(ab')2 fragments are bivalent fragments containing two Fab fragments linked together. (iii) an Fd fragment consisting of a VH domain and a CH1 domain; (iv) an Fv fragment consisting of the VL and VH domains of a single arm of an antibody and (v) dAb fragments containing a single variable domain (Ward et al. (1989) Na ture 341:544-546, Winter et al., incorporated herein by reference. (vi) isolated complementarity determining regions (CDs) Furthermore, the two domains of the Fv fragment, VL and VH, are , which are encoded by separate genes, but which can be engineered to separate the VL and VH regions using recombinant methods. Single protein chains (known as single-chain Fv (scFv)) pair to form monovalent molecules. For example, Bird et al. (1988) Science 242:423-426 and Huston et al. (1988) Proc. Natl. Acad. Sci. USA 8 5:5879-5883). Such single chain antibodies are also encompassed within the term "antigen-binding portion" of an antibody. In certain embodiments of the invention, the scFv molecule is Other forms of single chain antibodies, such as diabodies, may also be included. are bivalent, bispecific antibodies in which the VH and VL domains are expressed on a single polypeptide chain. antibody, but with a linker that is too short to allow the two domains to pair on the same chain. The use of anchors encourages domains to pair with complementary domains on other strands. to create two antigen-binding sites (see, e.g., Holliger, P. et al. (1993) )Proc.Natl.Acad.Sci.USA 90 :6444-6448, Pol See Jak, RJ et al. (1994) Structure 2:1121-1123 Such antibody binding moieties are known in the art (Kontermann and Antibody Engineering (ed.) Dubel (2001) Springer Verlag New York, 790pp (ISBN 3-540-41354-5) .

[0045] "Fully human" antibodies are derived from human immunoglobulins (e.g., human immunoglobulin codons). The term "human antibody" includes antibody variable domains having sequences (derived from humanized sequences). The term refers to, for example, variable and constant regions derived from human germline immunoglobulin sequences (e.g., As used herein, antibodies or fragments such as variable domains include antibodies having a variable domain (where present). The term "human" as applied to a segment refers to the incorporation of human constant region sequences into an antibody polypeptide. by grafting (i.e., replacing non-human constant regions with human constant regions), or by Grafting of human V region framework sequences from mammalian to immunoglobulin variable domains (e.g. That is, by replacing the non-human framework regions of the V domain with human framework regions. This does not include antibodies from another species, e.g., mouse, that have been "humanized" by complementarity determination. A method for humanizing immunoglobulin variable regions through rational modification of residues is described (US2006 / 0258852).

[0046] The term "humanized antibody" refers to an antibody that has one or more complementarity determining regions (CDRs) from a non-human species. ) and antibodies from non-human species having framework regions from human immunoglobulin molecules. A humanized antibody optionally contains one or more fragments derived from the non-human species from which the CDRs are derived. Such framework sequences may further comprise framework residues. from public DNA databases covering all gene sequences or from published references. For example, the germline DNA sequences of human heavy and light chain variable region genes can be obtained by "Base" Human Germline Sequence Database (www.mrccpe.com.ac. Available on the web at uk / vbase and also at Kaba t, EA et al., 1991, Sequences of Proteins of Immu Immunogens To avoid loss of activity during the degradation process, the variable region framework sequences of human antibodies are at least It undergoes back mutation and maintains activity.

[0047] Humanized antibodies may be of any class of immune response, including IgM, IgG, IgD, IgA, and IgE. Globulins, including but not limited to IgG1, IgG2, IgG3, and IgG4 Humanized antibodies can be selected from any isotype not listed in Table 1. Humanized antibodies can be selected from any isotype not listed in Table 1. The specific constant domains may include sequences from any isotype, and are well known in the art. The desired effector functions may be optimized using techniques such as those described above.

[0048] The term "multispecific" antibody refers to an antibody that is directed against two or more different epitopes in a single antibody molecule. In addition to the standard antibody structure, other binding molecules have binding domains that bind to two Bispecific or multispecific antibodies can be constructed with two binding specificities. Tope binding can be simultaneous or sequential. Triomas and Hybrid Hybridomas are two examples of cell lines capable of secreting bispecific antibodies. Bispecific antibodies can also be produced by recombinant (Strohlein and Heiss, Futur re Oncol.6:1387-94(2010), Mabry and Snavely, IDrugs. 13:543-9(2010)). Bispecific antibodies are also known as diabodies. It could be.

[0049] As used herein, the term "labeled antibody" refers to a binding protein, e.g., an antibody It refers to an antibody or antigen-binding portion thereof incorporating a label that provides identification of the antibody or antigen-binding portion thereof. Identification can be achieved by the incorporation of a detectable marker, e.g., a radiolabeled amino acid, or by the incorporation of a marked amino acid. avidin (e.g., containing a fluorescent marker or enzymatic activity that can be detected by optical or colorimetric methods) The attachment of a biotinyl moiety to a polypeptide that can be detected by biotinylation (e.g., streptavidin). Examples of labels for polypeptides include radioisotopes or radionuclides (e.g., 3 H, 1 4 C. 35 S, 90 Y, 99 Tc, 111 In, 125 I, 131 I,177 Lu, 16 6 Ho or 153 Sm) and fluorescent labels (e.g., FITC, rhodamine, lanthanide ligands) fluorescent dyes), enzyme labels (e.g., horseradish peroxidase, luciferase, alkaline Phosphatase), a chemiluminescent marker, a biotinyl group, and a secondary reporter. The polypeptide epitope to be recognized (e.g., leucine zipper pair sequence, binding site, metal binding domain, epitope tag) and magnetic agents such as gadolinium chelates and, but are not limited to these.

[0050] A "conservative amino acid substitution" is one in which one amino acid is replaced with another amino acid having a similar side chain. Families of amino acids with similar side chains are defined in the art as and amino acids with basic side chains (e.g., lysine, arginine, histidine) , amino acids with acidic side chains (e.g., aspartic acid, glutamic acid), uncharged polar side Chain-forming amino acids (e.g., asparagine, glutamine, serine, threonine, tyrosine, amino acids with non-polar side chains (e.g., glycine, alanine, valine, , leucine, isoleucine, proline, phenylalanine, methionine, tryptophan ), amino acids with β-branched side chains (e.g., threonine, valine, isoleucine) and aromatic Amino acids with aromatic side chains (e.g., tyrosine, phenylalanine, tryptophan, hydroxybenzoates, For example, substituting tyrosine with phenylalanine is a conservative substitution. In certain embodiments, conservative sequences in the sequences of the polypeptides and antibodies of the present disclosure are The substitution prevents binding of an antibody containing the polypeptide or amino acid sequence to the antigen to which the binding molecule binds. Methods for identifying conservative nucleotide and amino acid substitutions that do not eliminate antigen binding. Methods are well known in the art (see, for example, Brummell et al., Biochem. 32: 1180-1187(1993), Kobayashi et al., Protein Eng.1 2(10):879-884 (1999) and Burkset et al., Proc. Natl. Acad. Sci. USA 94:412-417 (1997).

[0051] The term "polynucleotide" is intended to include single nucleic acids and multiple nucleic acids, Isolated nucleic acid molecules or constructs, e.g., messenger RNA (mRNA), cDNA or plasmid DNA (pDNA). Polynucleotides are amide bonds or non-conventional bonds (e.g., those found in peptide nucleic acids (PNA)) The term "nucleic acid" or "nucleic acid sequence" refers to a polynucleotide. Any one or more nucleic acid segments, e.g., DNA or RNA fragments, present in Refers to...

[0052] An "isolated" nucleic acid or polynucleotide is any nucleic acid or polynucleotide that is separated from its native environment. For example, gel-purified polynucleotides are intended to be nucleic acids or polynucleotides in the form of A recombinant polynucleotide encoding a polypeptide contained in a peptide or vector is a "single It is also thought to have been engineered to have restriction sites for cloning. A polynucleotide segment obtained, e.g., a PCR product, is considered to be "isolated." Further examples of isolated polynucleotides include recombinant polynucleotides maintained in heterologous host cells. oligonucleotides, or in non-natural solutions such as buffers or saline solutions (partially or substantially ) purified polynucleotides. Isolated RNA molecules include polynucleotides. In vivo or in vitro RNA transcripts of the peptides are included, where the transcripts are not those found in nature. An isolated polynucleotide or nucleic acid is not one that has been produced synthetically. Furthermore, the polynucleotide or nucleic acid may further comprise a molecule such as a promoter, a ribosomal It may be or contain regulatory elements such as a nucleotide binding site or a transcription terminator. good.

[0053] As used herein, the term "expression" refers to the process by which a gene is produced as a biochemical, e.g., a polypeptide. This refers to a process for producing a polypeptide, including, but not limited to, gene knockdown. Any expression of a gene in a cell, including both transient and stable expression, as well as the functional presence of a gene in a cell. This process includes, but is not limited to, the transcription of genes into messenger RNA (mRNA). This involves transcription of the gene and translation of such mRNA into a polypeptide. If the product is a biochemical, expression includes the production of that biochemical and any precursors. Expression of a gene produces a "gene product." As used herein, A gene product is a nucleic acid, e.g., messenger RNA produced by transcription of a gene, or a polypeptide translated from a transcription product. The gene product may be a nucleic acid with post-transcriptional modifications, such as polyadenylation, or a post-translational modification, For example, methylation, glycosylation, lipid addition, binding to other protein subunits, Further included are polypeptides with proteolytic cleavage and the like.

[0054] As used herein, a "neutralizing antibody" (e.g., an antibody that inhibits NAMPT activity) is used. The term "antibodies that inhibit NAMPT" includes antibodies that inhibit the biological activity of NAMPT by binding to NAMPT. Neutralizing antibodies are antibodies that reduce the amount of binding partner that binds to the determinant by excess antibody. For example, as measured by target molecule activity or in an in vitro competitive binding assay. So at least about 20%, 30%, 40%, 50%, 60%, 70%, 80%, 85% , 90%, 95%, 97%, 99% or more reduction of the ligand or substrate As will be appreciated, the modified activity is a function of the activity of a compound of the present invention. The activity may be measured directly using art-recognized techniques or the altered activity may be measured below. The biological activity of NAMPT or its ligands may be measured by its effect on the flow of This inhibition of NAMPT levels (either in vitro or in vivo) is related to the amount of extracellular NAMPT. T-induced cell activation (e.g., NFκB phosphorylation) and NAMP to NAMPT ligands NAMPT biological activity is assessed by measuring one or more indicators of NAMPT biological activity, such as T binding. These indicators of NAMPT biological activity are known in the art. It can be assessed by one or more standard in vitro or in vivo assays (implementation For example, in some embodiments, the antibody is capable of inhibiting NAMPT activity. Potency is assessed by the inhibition of NAMPT-induced activation of endothelial cells. As an additional or alternative parameter, the antibody may be used to measure NAMPT-induced cell activation. The ability to inhibit NAMPT-induced transcriptional activity via FκB can be assessed.

[0055] "Treat" or "Treatment" or "To treat" or "Alleviate" or "To alleviate" " and the like means to cure, slow, or alleviate the symptoms of an existing diagnosed pathological disease or disorder, and / or treatments that halt or slow the progression of the disease. Such treatments include "Prevent" includes, but is not required to, the complete removal of symptoms or cure of a disease. The terms "avoid," "prevent," and "prevent" are used to refer to the targeted undiagnosed pathological condition or disorder. Therefore, it refers to a preventative or preventative measure to prevent the onset of a disease. The definition of disability is given to people who already have a disability, who are vulnerable to a disability, who are at risk of developing a disability, and who are This may include people who prevent harm.

[0056] "Subject" or "individual" or "animal" or "patient" or "mammal" means a subject for diagnosis, prognosis, or any subject for whom treatment is desired, particularly a mammalian subject. Mammalian subjects include humans , domestic animals, farm animals, or dogs, cats, guinea pigs, rabbits, rats, mice, horses, pigs Animals include zoo animals, athletic animals or pet animals such as tigers, cows, bears etc.

[0057] As used herein, the terms "subject who will benefit from treatment" and "subject in need of treatment" refer to Phrases such as "animals that are resistant to NAMPT" and "animals that are resistant to NAMPT" refer to animals that are resistant to NAMPT from the administration of an anti-NAMPT antibody (e.g., a humanized anti-NAMPT antibody). Such antibodies may be used to treat, for example, diseases, e.g., For use in diagnostic procedures and / or treatment or prevention of inflammatory lung disorders or cancer (e.g., prostate cancer). It can be used.

[0058] The terms "effective amount" and "therapeutically effective amount" are used interchangeably herein, and in particular The compounds, formulations, materials or compositions described herein are effective to achieve a specific biological result. Such results include inhibition of NAMPT expression or activity, or the amount of a composition that is of signaling molecules downstream of NAMPT, as determined by any means appropriate in the field. For example, NAMPT activity may include, but is not limited to, the expression or activity of: Cytokine activity, nicotinamide phosphoribosyltransferase activity, chemotactic factors , NF-κB signaling activity, redox signaling activity and / or mitochondria These include, but are not limited to, roles in cell function and apoptosis. In addition, with respect to a therapeutically effective amount, one skilled in the art would understand that such an amount would be considered a therapeutically effective amount. Even if a drug is administered, it may not always be effective in treating the disease. Amounts, drug delivery amounts, therapeutically effective amounts, and therapeutic levels are used herein for adult human subjects. Those skilled in the art will appreciate that the specific therapeutic agents needed to treat a particular subject and / or disease are provided. Such amounts can be adjusted according to standard practices.

[0059] Anti-NAMPT antibody As provided herein, the present disclosure provides an anti-nicotinamide phosphoribosyltransferase. Antibodies to NAMPT-related enzymes or antigen-binding fragments thereof For prophylactic and therapeutic use in patients suffering from local and systemic inflammatory disorders and compositions, methods, and articles of manufacture (e.g., kits) comprising the antibodies or antigen-binding fragments thereof. Nucleic acids comprising polynucleotide sequences encoding such antibodies are also included. In certain embodiments, the monoclonal antibodies provided herein The antibody binds to extracellular NAMPT (eNAMPT) and toll-like receptor 4 (TLR4). By preventing the activation of one or more downstream signaling pathways, and consequently and reducing or blocking systemic and pulmonary inflammation in certain respiratory disorders.

[0060] The NAMPT gene product converts mammalian nicotinamide to nicotinamide mononucleotide to enable nicotinamide adenine dinucleotide (NAD+) biosynthesis It is the rate-limiting enzyme in the D+ salvage pathway. The mature form of the extracellular NAMPT protein is approximately 120 It is a homodimer of 100 kDa (Takahashi et al., J. Biochem. 147:95 Mutations that reduce or inhibit the function of the NAMPT enzyme are associated with leukemia. and reducing the pathophysiological processes that lead to disorders such as pulmonary arterial hypertension (PAH). It has been established that this may be the case.

[0061] The human NAMPT gene (NAMPT) is located on chromosome 7 (segment 7q 22.3, base pairs 106248285 to 106286326). Human NAMPT gene The nucleic acid sequence of the product is known in the art, e.g., NCBI Reference Sequence: NM_00 5746.2, Homo sapiens nicotinamide phosphoribosyltransferase (NA MPT), see mRNA (Samal et al., Mol. Cell. Biol. 14(2 ), 1431-1437 (1994). The amino acid sequence of the human NAMPT enzyme is Known in the art, see, e.g., GenBank Accession No. NP00573 See 7.1. NAMPT binds to CD14+ In monocytes, macrophages, and dendritic cells Increases the production of IL-6, TNF-α and IL-Iβ, improving the effectiveness of T cells and B It has been shown to be involved in the development of both lymphocytes and T lymphocytes (Sun et al., Cyt Okine&Growth Factor Reviews 24(5):433-44 2 (2013)). The crystal structure of the NAMPT enzyme was reported by Kim et al., J. Mol. Biol., 362:66-77 (2006).

[0062] The receptor for NAMPT is Toll-like receptor 4 (TLR4), and in humans, TLR4 TLR4 is a protein encoded by a gene. It is a transmembrane protein, Members of the Toll-like receptor family, which belong to the pattern recognition receptor (PRR) family Activation of this receptor mediates the intracellular NF-κB signaling pathway and the activation of the innate immune system. Many gram-negative bacteria (e.g., Na It recognizes lipopolysaccharide (LPS), a component present in the Iseria species, and selects for Gram-positive bacteria. The receptor is best known for its ability to bind to the mitochondrial nuclei, which are the ligands for several viral proteins. Proteins, polysaccharides and low-density lipoproteins, beta-defensins, heat shock proteins, etc. The human TLR4 gene (TLR4) is located on chromosome 9 (SEQ ID NO: 1). It is located in segment 9q32-q33 (Georgel et al., PLoS ONE 4 (11):e7803(2009)). The nucleic acid sequence of the human TLR4 gene product is known in the art. For example, NCBI reference sequence: AAY82268.1, Homo sapiens Toll-like receptor 4 (TLR4), mRNA. The amino acid sequence of human TLR4 is Known in the art, see, e.g., GenBank Accession No. AAY8226 See 8.

[0063] In certain aspects, the present disclosure provides a humanized 1076 anti-hNAMPT antibody or its antigen-binding Examples of anti-NAMPT antibodies are provided below.

[0064] Regarding the anti-NAMPT antibody D-1076, D-1076 has (i) the amino acid sequence shown in SEQ ID NO: 3. a CDR1 domain having the amino acid sequence shown in SEQ ID NO:4; a CDR2 domain and a CDR3 domain having the amino acid sequence shown in SEQ ID NO:5 (ii) a heavy chain variable region having the amino acid sequence set forth in SEQ ID NO: 6, a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 7; and a CDR3 domain having the amino acid sequence shown in SEQ ID NO: 8. and a light chain variable region having the amino acid sequence set forth in SEQ ID NO:2, including a nucleotide sequence identical to that set forth in SEQ ID NO:3.

[0065] Regarding the anti-NAMPT antibody G-1076, G-1076 has (i) the amino acid sequence shown in SEQ ID NO: 3. a CDR1 domain having the amino acid sequence shown in SEQ ID NO:4; a CDR2 domain and a CDR3 domain having the amino acid sequence shown in SEQ ID NO:5 (ii) a heavy chain variable region having the amino acid sequence set forth in SEQ ID NO: 1; CDR1 domain having the amino acid sequence shown in SEQ ID NO: 1, a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 8; a light chain variable region having the amino acid sequence set forth in SEQ ID NO: 10, which includes three domains; do.

[0066] Regarding the anti-NAMPT antibody K-1076, K-1076 has (i) the amino acid sequence shown in SEQ ID NO: 3. a CDR1 domain having the amino acid sequence shown in SEQ ID NO:4; a CDR2 domain and a CDR3 domain having the amino acid sequence shown in SEQ ID NO:5 (ii) a heavy chain variable region having the amino acid sequence set forth in SEQ ID NO: 1; CDR1 domain having the amino acid sequence shown in SEQ ID NO: 1, the amino acid sequence shown in SEQ ID NO: 14 a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 8; a light chain variable region having the amino acid sequence set forth in SEQ ID NO: 13, which includes three domains; do.

[0067] Regarding the anti-NAMPT antibody N-1076, N-1076 has (i) the amino acid sequence shown in SEQ ID NO: 3. a CDR1 domain having the amino acid sequence shown in SEQ ID NO:4; a CDR2 domain and a CDR3 domain having the amino acid sequence shown in SEQ ID NO:5 (ii) a heavy chain variable region having the amino acid sequence set forth in SEQ ID NO: 15, a CDR1 domain having the amino acid sequence shown in SEQ ID NO:6, a CDR2 domain having the amino acid sequence shown in SEQ ID NO:7, a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 8; and a CDR3 domain having the amino acid sequence shown in SEQ ID NO: 8. a light chain variable region having the amino acid sequence set forth in SEQ ID NO:2, including a .

[0068] Regarding the anti-NAMPT antibody P-1076, P-1076 has the following structure: (i) a sequence shown in SEQ ID NO: 3; a CDR1 domain having the amino acid sequence shown in SEQ ID NO:4; a CDR2 domain and a CDR3 domain having the amino acid sequence shown in SEQ ID NO:5 (ii) a heavy chain variable region having the amino acid sequence set forth in SEQ ID NO: 15, 11, a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 14, a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 8; a light chain variable region having the amino acid sequence set forth in SEQ ID NO: 13, including an R3 domain; Has.

[0069] Regarding the anti-NAMPT antibody V-1076, V-1076 has (i) the amino acid sequence shown in SEQ ID NO: 3. a CDR1 domain having the amino acid sequence shown in SEQ ID NO:4; a CDR2 domain and a CDR3 domain having the amino acid sequence shown in SEQ ID NO:5 (ii) a heavy chain variable region having the amino acid sequence set forth in SEQ ID NO: 16, 11, a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 12, a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 8; a light chain variable region having the amino acid sequence set forth in SEQ ID NO: 10, including an R3 domain; Has.

[0070] Regarding the anti-NAMPT antibody X-1076, X-1076 has the following structure: (i) the amino acid sequence shown in SEQ ID NO: 3 a CDR1 domain having the amino acid sequence shown in SEQ ID NO:4; a CDR2 domain and a CDR3 domain having the amino acid sequence shown in SEQ ID NO:5 (ii) a heavy chain variable region having the amino acid sequence set forth in SEQ ID NO: 16, a CDR1 domain having the amino acid sequence shown in SEQ ID NO:6, a CDR2 domain having the amino acid sequence shown in SEQ ID NO:7, a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 8; and a CDR3 domain having the amino acid sequence shown in SEQ ID NO: 8. a light chain variable region having the amino acid sequence set forth in SEQ ID NO:2, including a .

[0071] Regarding the anti-NAMPT antibody P-1076-mod1, P-1076-mod1 is (i ) a CDR1 domain having the amino acid sequence shown in SEQ ID NO:3, a CDR2 domain having the amino acid sequence shown in SEQ ID NO:29, and a CDR2 domain having the amino acid sequence shown in SEQ ID NO:5. a heavy chain variable region having the amino acid sequence set forth in SEQ ID NO: 28, including a CDR3 domain and (ii) a CDR1 domain having the amino acid sequence set forth in SEQ ID NO: 11, SEQ ID NO: a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 14, and an amino acid sequence shown in SEQ ID NO: 8; The CDR3 domain has the amino acid sequence shown in SEQ ID NO: 30. and a light chain variable region corresponding to

[0072] Regarding the anti-NAMPT antibody P-1076-mod2, P-1076-mod2 is (i ) a CDR1 domain having the amino acid sequence shown in SEQ ID NO:3, a CDR2 domain having the amino acid sequence shown in SEQ ID NO:29, and a CDR2 domain having the amino acid sequence shown in SEQ ID NO:5. a heavy chain variable region having the amino acid sequence set forth in SEQ ID NO: 28, including a CDR3 domain and (ii) a CDR1 domain having the amino acid sequence set forth in SEQ ID NO: 11, SEQ ID NO: a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 14, and an amino acid sequence shown in SEQ ID NO: 8; The CDR3 domain has the amino acid sequence shown in SEQ ID NO: 31. and a light chain variable region corresponding to

[0073] Regarding the anti-NAMPT antibody P-1076-mod3, P-1076-mod3 is (i ) a CDR1 domain having the amino acid sequence shown in SEQ ID NO:3, a CDR2 domain having the amino acid sequence shown in SEQ ID NO:29, and a CDR2 domain having the amino acid sequence shown in SEQ ID NO:5. a heavy chain variable region having the amino acid sequence set forth in SEQ ID NO: 28, including a CDR3 domain and (ii) a CDR1 domain having the amino acid sequence set forth in SEQ ID NO: 11, SEQ ID NO: a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 33, and an amino acid sequence shown in SEQ ID NO: 8; The CDR3 domain has the amino acid sequence shown in SEQ ID NO: 32. and a light chain variable region corresponding to

[0074] Regarding the anti-NAMPT antibody P-1076-mod4, P-1076-mod4 is (i ) a CDR1 domain having the amino acid sequence shown in SEQ ID NO:3, a CDR2 domain having the amino acid sequence shown in SEQ ID NO:29, and a CDR2 domain having the amino acid sequence shown in SEQ ID NO:5. a heavy chain variable region having the amino acid sequence set forth in SEQ ID NO: 28, including a CDR3 domain and (ii) a CDR1 domain having the amino acid sequence set forth in SEQ ID NO: 11, SEQ ID NO: a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 35, and an amino acid sequence shown in SEQ ID NO: 8; The CDR3 domain has the amino acid sequence shown in SEQ ID NO: 34. and a light chain variable region corresponding to

[0075] Regarding the anti-NAMPT antibody P-1076-mod5, P-1076-mod5 is (i ) a CDR1 domain having the amino acid sequence shown in SEQ ID NO:3, a CDR2 domain having the amino acid sequence shown in SEQ ID NO:29, and a CDR2 domain having the amino acid sequence shown in SEQ ID NO:5. a heavy chain variable region having the amino acid sequence set forth in SEQ ID NO: 28, including a CDR3 domain and (ii) a CDR1 domain having the amino acid sequence set forth in SEQ ID NO: 11, SEQ ID NO: No. 37, and a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 8. The CDR3 domain has the amino acid sequence shown in SEQ ID NO: 36. and a light chain variable region corresponding to

[0076] Regarding the anti-NAMPT antibody P-1076-mod6, P-1076-mod6 is (i ) a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 3, a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 4 a CDR2 domain having the amino acid sequence shown in SEQ ID NO:5; a heavy chain variable region having the amino acid sequence set forth in SEQ ID NO: 15, including a CDR3 domain and (ii) a CDR1 domain having the amino acid sequence set forth in SEQ ID NO: 11, SEQ ID NO: 14, and a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 8. having the amino acid sequence set forth in SEQ ID NO: 30, including a CDR3 domain having the amino acid sequence and a light chain variable region comprising:

[0077] Regarding the anti-NAMPT antibody P-1076-mod7, P-1076-mod7 is (i ) a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 3, a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 4 a CDR2 domain having the amino acid sequence shown in SEQ ID NO:5; a heavy chain variable region having the amino acid sequence set forth in SEQ ID NO: 15, including a CDR3 domain and (ii) a CDR1 domain having the amino acid sequence set forth in SEQ ID NO: 11, SEQ ID NO: 14, and a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 8. having the amino acid sequence set forth in SEQ ID NO: 31, including a CDR3 domain having the amino acid sequence and a light chain variable region comprising:

[0078] Regarding the anti-NAMPT antibody P-1076-mod8, P-1076-mod8 is (i ) a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 3, a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 4 a CDR2 domain having the amino acid sequence shown in SEQ ID NO:5; a heavy chain variable region having the amino acid sequence set forth in SEQ ID NO: 15, including a CDR3 domain and (ii) a CDR1 domain having the amino acid sequence set forth in SEQ ID NO: 11, SEQ ID NO: 33, and a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 8. having the amino acid sequence set forth in SEQ ID NO: 32, including a CDR3 domain having the amino acid sequence and a light chain variable region comprising:

[0079] Regarding the anti-NAMPT antibody P-1076-mod9, P-1076-mod9 is (i ) a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 3, a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 4 a CDR2 domain having the amino acid sequence shown in SEQ ID NO:5; a heavy chain variable region having the amino acid sequence set forth in SEQ ID NO: 15, including a CDR3 domain and (ii) a CDR1 domain having the amino acid sequence set forth in SEQ ID NO: 11, SEQ ID NO: 35, and a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 8. having the amino acid sequence set forth in SEQ ID NO: 34, including a CDR3 domain having the amino acid sequence and a light chain variable region comprising:

[0080] Regarding the anti-NAMPT antibody P-1076-mod10, P-1076-mod10 is: (i) a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 3, a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 4 and a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 5. a heavy chain variable region having the amino acid sequence set forth in SEQ ID NO: 15, including a CDR3 domain having and (ii) a CDR1 domain having the amino acid sequence set forth in SEQ ID NO: 11, sequence a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 37, and a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 8 The amino acid sequence shown in SEQ ID NO: 36, including a CDR3 domain having the amino acid sequence and a light chain variable region having

[0081] Regarding the anti-NAMPT antibody P-1076-mod11, P-1076-mod11 is: (i) a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 3, a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 29 and a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 5. a heavy chain variant having the amino acid sequence set forth in SEQ ID NO: 28, which includes a CDR3 domain having the amino acid sequence (ii) a CDR1 domain having the amino acid sequence set forth in SEQ ID NO: 11; A CDR2 domain having the amino acid sequence shown in sequence number 14, and a CDR2 domain having the amino acid sequence shown in sequence number 8 the amino acid sequence shown in SEQ ID NO: 13, including a CDR3 domain having an amino acid sequence and a light chain variable region having:

[0082] In certain aspects, the present disclosure provides a humanized 1093 anti-hNAMPT antibody or its antigen-binding An example of the 1093 anti-NAMPT antibody fragment is shown below.

[0083] Regarding the anti-NAMPT antibody FF-1093, FF-1093 has the structure (i) of SEQ ID NO: 19. CDR1 domain having the amino acid sequence shown, amino acid sequence shown in SEQ ID NO: 20 and a CDR3 domain having the amino acid sequence shown in SEQ ID NO: 21. a heavy chain variable region having the amino acid sequence set forth in SEQ ID NO: 17, including the domain (ii ) a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 6, a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 7 a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 22; a light chain variable region having the amino acid sequence set forth in SEQ ID NO: 18, including a CDR3 domain It has a region.

[0084] Regarding the anti-NAMPT antibody II-1093, II-1093 has the structure (i) of SEQ ID NO: 19 CDR1 domain having the amino acid sequence shown, amino acid sequence shown in SEQ ID NO: 20 and a CDR3 domain having the amino acid sequence shown in SEQ ID NO: 21. a heavy chain variable region having the amino acid sequence set forth in SEQ ID NO: 23, including the domain (ii ) a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 6, a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 7 a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 22; a light chain variable region having the amino acid sequence set forth in SEQ ID NO: 24, including a CDR3 domain It has a region.

[0085] Regarding the anti-NAMPT antibody NN-1093, NN-1093 has the structure: (i) SEQ ID NO: 19 CDR1 domain having the amino acid sequence shown, amino acid sequence shown in SEQ ID NO: 20 and a CDR3 domain having the amino acid sequence shown in SEQ ID NO: 21. a heavy chain variable region having the amino acid sequence set forth in SEQ ID NO: 25, including the domain (ii ) a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 6, a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 7 a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 22; a light chain variable region having the amino acid sequence set forth in SEQ ID NO: 24, including a CDR3 domain It has a region.

[0086] Regarding the anti-NAMPT antibody PP-1093, PP-1093 has the structure: (i) SEQ ID NO: 19 CDR1 domain having the amino acid sequence shown, amino acid sequence shown in SEQ ID NO: 20 and a CDR3 domain having the amino acid sequence shown in SEQ ID NO: 21. a heavy chain variable region having the amino acid sequence set forth in SEQ ID NO: 25, including the domain (ii ) a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 6, a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 7 a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 22; a light chain variable region having the amino acid sequence set forth in SEQ ID NO: 18, including a CDR3 domain It has a region.

[0087] Regarding the anti-NAMPT antibody SS-1093, SS-1093 has the structure (i) of SEQ ID NO: 19 CDR1 domain having the amino acid sequence shown, amino acid sequence shown in SEQ ID NO: 20 and a CDR3 domain having the amino acid sequence shown in SEQ ID NO: 21. a heavy chain variable region having the amino acid sequence set forth in SEQ ID NO: 26, including the domain (ii ) a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 6, a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 7 a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 22; a light chain variable region having the amino acid sequence set forth in SEQ ID NO: 24, including a CDR3 domain It has a region.

[0088] Regarding the anti-NAMPT antibody UU-1093, UU-1093 has the structure (i) of SEQ ID NO: 19. CDR1 domain having the amino acid sequence shown, amino acid sequence shown in SEQ ID NO: 20 and a CDR3 domain having the amino acid sequence shown in SEQ ID NO: 21. a heavy chain variable region having the amino acid sequence set forth in SEQ ID NO: 26, including the domain (ii ) a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 6, a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 7 a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 22; a light chain variable region having the amino acid sequence set forth in SEQ ID NO: 18, including a CDR3 domain It has a region.

[0089] Regarding the anti-NAMPT antibody XX-1093, XX-1093 has the structure (i) of SEQ ID NO: 19. CDR1 domain having the amino acid sequence shown, amino acid sequence shown in SEQ ID NO: 20 and a CDR3 domain having the amino acid sequence shown in SEQ ID NO: 21. a heavy chain variable region having the amino acid sequence set forth in SEQ ID NO: 27, including the domain (ii ) a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 6, a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 7 a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 22; a light chain variable region having the amino acid sequence set forth in SEQ ID NO: 24, including a CDR3 domain It has a region.

[0090] Regarding the anti-NAMPT antibody ZZ-1093, ZZ-1093 has the structure (i) of SEQ ID NO: 19. CDR1 domain having the amino acid sequence shown, amino acid sequence shown in SEQ ID NO: 20 and a CDR3 domain having the amino acid sequence shown in SEQ ID NO: 21. a heavy chain variable region having the amino acid sequence set forth in SEQ ID NO: 27, including the domain (ii ) a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 6, a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 7 a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 22; a light chain variable region having the amino acid sequence set forth in SEQ ID NO: 18, including a CDR3 domain It has a region.

[0091] Regarding the anti-NAMPT antibody UU-1093-mod1, UU-1093-mod1 is (i) a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 19, a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 39 CDR2 domain having the amino acid sequence shown, and the amino acid shown in SEQ ID NO: 21 a heavy chain having the amino acid sequence set forth in SEQ ID NO: 38, including a CDR3 domain having the sequence a chain variable region and (ii) a CDR1 domain having the amino acid sequence set forth in SEQ ID NO: 11. a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 7, and a CDR3 domain having the amino acid sequence shown in SEQ ID NO: 22 47, which contains a CDR3 domain having an amino acid sequence as set forth in SEQ ID NO: 47. and a light chain variable region having the sequence:

[0092] Regarding the anti-NAMPT antibody UU-1093-mod2, UU-1093-mod2 is (i) a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 19, a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 39 CDR2 domain having the amino acid sequence shown, and the amino acid shown in SEQ ID NO: 41 a heavy chain having the amino acid sequence set forth in SEQ ID NO: 40, including a CDR3 domain having the sequence a chain variable region and (ii) a CDR1 domain having the amino acid sequence set forth in SEQ ID NO: 11. a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 7, and a CDR3 domain having the amino acid sequence shown in SEQ ID NO: 22 47, which contains a CDR3 domain having an amino acid sequence as set forth in SEQ ID NO: 47. and a light chain variable region having the sequence:

[0093] Regarding the anti-NAMPT antibody UU-1093-mod3, UU-1093-mod3 is: (i) a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 43, a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 39 CDR2 domain having the amino acid sequence shown, and the amino acid shown in SEQ ID NO: 44 a heavy chain having the amino acid sequence set forth in SEQ ID NO: 42, including a CDR3 domain having the sequence a chain variable region and (ii) a CDR1 domain having the amino acid sequence set forth in SEQ ID NO: 11. a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 7, and a CDR3 domain having the amino acid sequence shown in SEQ ID NO: 22 47, which contains a CDR3 domain having an amino acid sequence as set forth in SEQ ID NO: 47. and a light chain variable region having the sequence:

[0094] Regarding the anti-NAMPT antibody UU-1093-mod4, UU-1093-mod4 is (i) a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 46, a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 39 CDR2 domain having the amino acid sequence shown, and the amino acid shown in SEQ ID NO: 41 a heavy chain having the amino acid sequence set forth in SEQ ID NO: 45, including a CDR3 domain having the sequence a chain variable region and (ii) a CDR1 domain having the amino acid sequence set forth in SEQ ID NO: 11. a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 7, and a CDR3 domain having the amino acid sequence shown in SEQ ID NO: 22 47, which contains a CDR3 domain having an amino acid sequence as set forth in SEQ ID NO: 47. and a light chain variable region having the sequence:

[0095] Regarding the anti-NAMPT antibody UU-1093-mod5, UU-1093-mod5 is: (i) a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 19, a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 39 CDR2 domain having the amino acid sequence shown, and the amino acid shown in SEQ ID NO: 21 a heavy chain having the amino acid sequence set forth in SEQ ID NO: 38, including a CDR3 domain having the sequence a chain variable region and (ii) a CDR1 domain having the amino acid sequence set forth in SEQ ID NO: 49 , a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 50, and a CDR3 domain having the amino acid sequence shown in SEQ ID NO: 22 48, which contains a CDR3 domain having the amino acid sequence shown. and a light chain variable region having the sequence

[0096] Regarding the anti-NAMPT antibody UU-1093-mod6, UU-1093-mod6 is (i) a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 19, a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 39 CDR2 domain having the amino acid sequence shown, and the amino acid shown in SEQ ID NO: 41 a heavy chain having the amino acid sequence set forth in SEQ ID NO: 40, including a CDR3 domain having the sequence a chain variable region and (ii) a CDR1 domain having the amino acid sequence set forth in SEQ ID NO: 49 , a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 50, and a CDR3 domain having the amino acid sequence shown in SEQ ID NO: 22 48, which contains a CDR3 domain having the amino acid sequence shown. and a light chain variable region having the sequence

[0097] Regarding the anti-NAMPT antibody UU-1093-mod7, UU-1093-mod7 is: (i) a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 43, a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 39 CDR2 domain having the amino acid sequence shown, and the amino acid shown in SEQ ID NO: 44 a heavy chain having the amino acid sequence set forth in SEQ ID NO: 42, including a CDR3 domain having the sequence a chain variable region and (ii) a CDR1 domain having the amino acid sequence set forth in SEQ ID NO: 49 , a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 50, and a CDR3 domain having the amino acid sequence shown in SEQ ID NO: 22 48, which contains a CDR3 domain having the amino acid sequence shown. and a light chain variable region having the sequence

[0098] Regarding the anti-NAMPT antibody UU-1093-mod8, UU-1093-mod8 is (i) a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 46, a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 39 CDR2 domain having the amino acid sequence shown, and the amino acid shown in SEQ ID NO: 41 a heavy chain having the amino acid sequence set forth in SEQ ID NO: 45, including a CDR3 domain having the sequence a chain variable region and (ii) a CDR1 domain having the amino acid sequence set forth in SEQ ID NO: 49 , a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 50, and a CDR3 domain having the amino acid sequence shown in SEQ ID NO: 22 48, which contains a CDR3 domain having the amino acid sequence shown. and a light chain variable region having the sequence

[0099] Regarding the anti-NAMPT antibody UU-1093-mod9, UU-1093-mod9 is (i) a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 19, a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 39 CDR2 domain having the amino acid sequence shown, and the amino acid shown in SEQ ID NO: 21 a heavy chain having the amino acid sequence set forth in SEQ ID NO: 38, including a CDR3 domain having the sequence a chain variable region and (ii) a CDR1 domain having the amino acid sequence set forth in SEQ ID NO: 11. , a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 52, and a CDR3 domain having the amino acid sequence shown in SEQ ID NO: 53 51, which contains a CDR3 domain having the amino acid sequence shown. and a light chain variable region having the sequence

[0100] About the anti-NAMPT antibody UU-1093-mod10 (i) a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 19, a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 3 9, and a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 41. having the amino acid sequence set forth in SEQ ID NO: 40, including a CDR3 domain having the amino acid sequence and (ii) a heavy chain variable region having the amino acid sequence shown in SEQ ID NO: 11. a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 52, and a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 5 3, comprising a CDR3 domain having the amino acid sequence shown in SEQ ID NO: 51. and a light chain variable region having the amino acid sequence

[0101] About the anti-NAMPT antibody UU-1093-mod11 (i) a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 43, 9, and a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 44. having the amino acid sequence set forth in SEQ ID NO: 42, including a CDR3 domain having the amino acid sequence and (ii) a heavy chain variable region having the amino acid sequence shown in SEQ ID NO: 11. a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 52, and a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 5 3, comprising a CDR3 domain having the amino acid sequence shown in SEQ ID NO: 51. and a light chain variable region having the amino acid sequence

[0102] About the anti-NAMPT antibody UU-1093-mod12 (i) a CDR1 domain having the amino acid sequence set forth in SEQ ID NO: 46, 9, and a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 41. having the amino acid sequence set forth in SEQ ID NO: 45, including a CDR3 domain having the amino acid sequence and (ii) a heavy chain variable region having the amino acid sequence shown in SEQ ID NO: 11. a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 52, and a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 5 3, comprising a CDR3 domain having the amino acid sequence shown in SEQ ID NO: 51. and a light chain variable region having the amino acid sequence

[0103] About the anti-NAMPT antibody UU-1093-mod13 (i) a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 19, a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 3 9, and a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 21. having the amino acid sequence set forth in SEQ ID NO: 38, including a CDR3 domain having the amino acid sequence and (ii) a heavy chain variable region having the amino acid sequence shown in SEQ ID NO:6. a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 7, and a CDR3 domain having the amino acid sequence shown in SEQ ID NO: 22 18, which contains a CDR3 domain having the amino acid sequence shown. and a light chain variable region having the sequence

[0104] About the anti-NAMPT antibody UU-1093-mod14 (i) a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 19, a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 3 9, and a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 41. having the amino acid sequence set forth in SEQ ID NO: 40, including a CDR3 domain having the amino acid sequence and (ii) a heavy chain variable region having the amino acid sequence shown in SEQ ID NO:6. a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 7, and a CDR3 domain having the amino acid sequence shown in SEQ ID NO: 22 18, which contains a CDR3 domain having the amino acid sequence shown. and a light chain variable region having the sequence

[0105] About the anti-NAMPT antibody UU-1093-mod15 (i) a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 43, 9, and a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 44. having the amino acid sequence set forth in SEQ ID NO: 42, including a CDR3 domain having the amino acid sequence and (ii) a heavy chain variable region having the amino acid sequence shown in SEQ ID NO:6. a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 7, and a CDR3 domain having the amino acid sequence shown in SEQ ID NO: 22 18, which contains a CDR3 domain having the amino acid sequence shown. and a light chain variable region having the sequence

[0106] About the anti-NAMPT antibody UU-1093-mod16 (i) a CDR1 domain having the amino acid sequence set forth in SEQ ID NO: 46, 9, and a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 41. having the amino acid sequence set forth in SEQ ID NO: 45, including a CDR3 domain having the amino acid sequence and (ii) a heavy chain variable region having the amino acid sequence shown in SEQ ID NO:6. a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 7, and a CDR3 domain having the amino acid sequence shown in SEQ ID NO: 22 18, which contains a CDR3 domain having the amino acid sequence shown. and a light chain variable region having the sequence

[0107] About the anti-NAMPT antibody UU-1093-mod17 (i) a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 19, a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 21; having the amino acid sequence set forth in SEQ ID NO: 26, including a CDR3 domain having the amino acid sequence and (ii) a heavy chain variable region having the amino acid sequence shown in SEQ ID NO: 11. a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 7, and a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 22 47, which contains a CDR3 domain having the amino acid sequence shown in SEQ ID NO: 47. and a light chain variable region having the amino acid sequence

[0108] About the anti-NAMPT antibody UU-1093-mod18 (i) a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 19, a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 21; having the amino acid sequence set forth in SEQ ID NO: 26, including a CDR3 domain having the amino acid sequence and (ii) a heavy chain variable region having the amino acid sequence set forth in SEQ ID NO: 49. a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 50, and a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 2 2, comprising a CDR3 domain having the amino acid sequence shown in SEQ ID NO: 48. and a light chain variable region having the amino acid sequence

[0109] About the anti-NAMPT antibody UU-1093-mod19 (i) a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 19, a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 21; having the amino acid sequence set forth in SEQ ID NO: 26, including a CDR3 domain having the amino acid sequence and (ii) a heavy chain variable region having the amino acid sequence shown in SEQ ID NO: 11. a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 52, and a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 5 3, comprising a CDR3 domain having the amino acid sequence shown in SEQ ID NO: 51. and a light chain variable region having the amino acid sequence

[0110] In some embodiments, the present invention provides a method for the production of human NAMPT in the form of a NAMPT homodimer. Some of the anti-NAMPT antibodies bind to discontinuous epitopes on human NAMPT. In some embodiments, the antibody or antigen-binding portion thereof comprises amino acid residue 1 of SEQ ID NO: 60. at least one amino acid of 7 to 44, and amino acid residues 117 to 12 of SEQ ID NO: 60 7, and amino acid residues 162 to 170 of SEQ ID NO: 60. At least one amino acid among amino acid residues 242 to 261 of SEQ ID NO: 60 At least one amino acid, at least one amino acid from amino acid residues 262 to 273 of SEQ ID NO: 60 another amino acid, at least one of amino acid residues 289 to 305 of SEQ ID NO: 60; at least one amino acid selected from amino acid residues 332 to 342 of SEQ ID NO: 60; at least one amino acid selected from amino acid residues 374 to 389 of SEQ ID NO: 60; At least one amino acid among amino acid residues 418 to 425 of SEQ ID NO: 60, SEQ ID NO: At least one amino acid among amino acid residues 453 to 466 of SEQ ID NO: 60, and / or Human containing at least one amino acid from 408 to 416 of sequence number 60 In some embodiments, the antibody or its The antigen-binding portion comprises at least one amino acid residue from 29 to 51 of SEQ ID NO: 60. at least one amino acid selected from amino acid residues 61 to 72 of SEQ ID NO: 60; At least one amino acid among amino acid residues 156 to 170 of SEQ ID NO: 60, at least one amino acid of amino acid residues 216 to 234 of SEQ ID NO: 60; at least one amino acid of amino acid residues 316 to 331, amino acid of SEQ ID NO: 60 At least one amino acid from residues 332 to 342, amino acid residue 3 of SEQ ID NO: 60 at least one amino acid of 73 to 389 of SEQ ID NO: 60, amino acid residues 417 to 419 of SEQ ID NO: 60; at least one amino acid of 431, amino acid residues 454 to 469 of SEQ ID NO: 60 and / or amino acid residues 470-474 of SEQ ID NO: 60 Binds to an epitope on human NAMPT that contains at least one amino acid out of 78 .

[0111] In certain embodiments, the provided antibodies or fragments thereof are capable of detecting plasmon resonance signals. When measured by the method described above, the activity of human NAMPT expressed on human cells ranges from about 3 nM to about 2 nM. 0 nM binding affinity (K D ) can be included.

[0112] In certain embodiments, the antibodies or antigen-binding fragments thereof described herein , e.g., 5 x 10 -2 seconds -1 , 10 -2 seconds -1 , 5×10 -3 seconds -1 , 10 -3 seconds -1 , 5×10 -4 seconds -1 , 10 -4 seconds -1 , 5×10 -5 seconds -1 or 10 -5 seconds -1 , 5× 10 -6 seconds -1 , 10 -6 seconds -1 , 5×10 -7 seconds -1 or 10 -7 seconds -1 Dissociation rate of degree(k (off) ) and binds to NAMPT, for example, human NAMPT.

[0113] In certain embodiments, the antibodies or antigen-binding fragments thereof described herein , for example, for example, 10 3 M -1 seconds -1 , 5×10 3 M -1 seconds -1 , 104 M -1 seconds -1 , 5×10 4 M -1 seconds -1 , 10 5 M -1 seconds -1 , 5×10 5 M -1 seconds -1 , 10 6 M - 1 seconds -1 or 5 x 10 6 M -1 seconds -1 or 10 7 M -1 seconds -1 Association rate (k (on ) ) and binds to NAMPT, for example, human NAMPT.

[0114] In certain embodiments, the antibodies or antigen-binding fragments thereof described herein , e.g. 5×10 -7 M, 10 -7 M, 5 x 10 -8 M, 10 -8 M, 5 x 10 -9 M. 10 -9 M, 5 x 10 -10 M, 10 -10 M, 5 x 10 -11 M, 10 -11 M, 5× 10 -12 M, 10 -12 M or 5 x 10 -13 Dissociation constant or K D In NAMPT , for example, binds to human NAMPT. Binding affinity can be measured using various methods known in the art. techniques such as surface plasmon resonance (SPR), biolayer interferometry, dual-polarized interferometry, Static light scattering, dynamic light scattering, isothermal titration calorimetry, ELISA, analytical ultracentrifugation and flow The amount of erythrocyte mass may be determined using immunocytometry.

[0115] In certain embodiments, the anti-NAMPT antibody or antigen binding thereof provided herein The fragment may further comprise a heterologous agent, such as a stabilizer, immune response modifier, or detectable agent. In certain embodiments, the heterologous agent may be attached to the polypeptide via a peptide bond. One or more additional polypeptide sequences fused to the subunit, e.g., a signal sequence ( For example, a secretory signal sequence, a linker sequence, an amino acid tag or label, or a sequence that facilitates purification. In certain embodiments, the heterologous polypeptide may comprise a peptide or polypeptide sequence that The domain may be any heavy or light antibody subunit or its derivatives, as long as the functional characteristics of the domain are maintained. It can be fused to the N-terminus or C-terminus of either of these fragments.

[0116] In certain embodiments, the heterologous agent, as provided herein, is an anti-NAMPT antibody. The polypeptide subunits can be chemically linked to the polypeptide or antigen-binding fragments thereof. Exemplary heterologous agents that can be chemically attached to the unit include, but are not limited to, linkers, Drugs, toxins, contrast agents, radioactive compounds, organic and inorganic polymers, and polypeptides Any other composition that can provide a desired activity that the dosubunit itself does not provide. Specific agents include, but are not limited to, polyethylene glycol (PEG), cytotoxic These include agents, radionuclides, contrast agents, and biotin.

[0117] In some embodiments, the anti-NAMPT antibody or fragment is The antibodies disclosed herein are labeled with a radioactive label for use in in vivo detection. Examples of radioisotopes that may be used to identify211 , I 131 , I 1 25 , Y 90 ,Re 186 ,Re 188 , Sm 153 , Bi 212 , P 32 , Pb 212 or a radioactive isotope of Lu. In this regard, the present invention provides a method for the preparation of a radioactive compound, as described herein, which is bound to a radioactive atom to form a radioconjugate. A variety of radioisotopes are available for the preparation of radioconjugates, including anti-NAMPT antibodies. For example, At 211 , I 131 , I 125 , Y 90 ,Re 186 ,Re 188 , Sm 153 , Bi 212 , P 32 , Pb 212 , radioactive isotopes of Lu. The complexes, when used for detection, contain radioactive atoms, e.g., tc 99m or I123, or again iodine-123, iodine-131, indium-111, Fluorine-19, carbon-13, nitrogen-15, oxygen-17, gadolinium, manganese or iron Any scan for nuclear magnetic resonance (NMR) imaging (also known as magnetic resonance imaging, MRI) Pin indicators may also be included.

[0118] In certain embodiments, any of the anti-NAMPT antibodies provided herein As used herein, the term "antibody" is useful for detecting the presence of NAMPT in a biological sample. The term "detection" as used herein includes quantitative detection or qualitative detection. The biological sample may include cells or tissues such as cerebrospinal fluid, lung cells or tissues, or blood.

[0119] In some embodiments, anti-NAMPT antibodies for use in methods of diagnosis or detection In a further aspect, a method for detecting the presence of NAMPT in a biological sample is provided. In certain embodiments, the method comprises the step of: contacting the biological sample with an anti-NAMPT antibody as described herein under conditions that allow the and detecting whether a complex is formed between the anti-NAMPT antibody and NAMPT. Such a method may be an in vitro or an in vivo method. Furthermore, the complex formed between the anti-NAMPT antibody and NAMPT in the test biological sample is compared to the complex formed in a control biological sample (e.g., a biological sample from a healthy subject). The antibody formed between the anti-NAMPT antibody and NAMPT in the test biological sample can be compared. The amount of complex formed is quantified and compared to a control biological sample (e.g., a biological sample from a healthy subject). The amount of complex formed in the test subject or the average amount of complex known to be formed in healthy subjects It can also be compared with the average weight.

[0120] The anti-NAMPT antibodies and fragments disclosed herein also include antibodies against human NAMPT, and can be used in some embodiments as a reagent for detecting mouse NAMPT. For example, the anti-NAMPT antibodies described herein can be used in an ELISA assay. Detection of the presence of NAMPT may be performed using a wide variety of tissues and samples, including plasma or serum. Western blotting (with or without immunoprecipitation) to assay for Fluorescence-activated cell sorting (FACS), flow cytometry, and ELISA procedures This may be achieved in a number of ways using the antibodies and fragments disclosed herein. A wide variety of assay formats using such assays can be used that include the antibodies disclosed herein. Immunoassay techniques are available, see, for example, U.S. Pat. No. 4,016,043 ... See, for example, US Pat. Nos. 4,242,799 and 4,018,653. These assays include non-competitive Both single-site and two-site or "sandwich" assays, as well as conventional These assays involve competitive binding assays of a label to a target biomarker. Direct binding of anti-NAMPT antibodies is also included.

[0121] Sandwich assays are the most useful and commonly used assays. Many variations of the assay technique exist, all of which are intended to be encompassed by the present invention. Briefly, in a typical forward assay, unlabeled antibodies are immobilized on a solid substrate. The sample to be tested is contacted with the binding molecule. The sample is then contacted with the binding molecule for a period of time sufficient to allow the formation of antibody-antigen complexes. After incubation for a sufficient period of time, the cells are labeled with a reporter molecule that can generate a detectable signal. A second antibody specific to the antigen is added to form another antibody-antigen-labeled antibody complex. Incubate for a sufficient time to remove any unreacted material and allow the reporter molecule to react. The presence of antigen is determined by observing the signal generated. The method may be qualitative by observing the presence of a known amount of biomarker. The amount of the analyte present may be quantified by comparison with a control sample containing the analyte.

[0122] For recombinant production of an anti-NAMPT antibody, a nucleic acid encoding the antibody, e.g., as described above, can be prepared. and isolating the isolated gene and using one or more vectors for further cloning and / or expression in a host cell. Such nucleic acids can be inserted into the ribozyme using conventional procedures (e.g., to insert heavy and light chains of antibodies). Using oligonucleotide probes capable of specifically binding to the encoding gene. They may be easily isolated (by cloning) and sequenced.

[0123] Suitable host cells for cloning or expressing antibody-encoding vectors include those described herein. The described prokaryotic or eukaryotic cells include, for example, antibodies, particularly those containing glycosylated and F c If effector functions are not required, antibodies may be produced in bacteria. For expression of fragments and polypeptides, see, e.g., U.S. Pat. No. 5,648,237. , 5789199 and 5840523. (Antibody Flag in E. coli Charlton, Methods in Molecular Biology, Vol.248 (edited by BKCLo, Humana Press, Totowa, NJ, 2003), pp. 245-254. The organisms may be isolated from the bacterial cell paste in a soluble fraction and further purified.

[0124] In addition to prokaryotes, eukaryotic microbes such as filamentous fungi or yeast can also be used to host antibody-encoding vectors. and the glycosylation pathway has been "humanized." As a result, fungal and yeast strains that produce antibodies with partially or completely human glycosylation patterns Gerngross, Nat. Biotech. 22:1409-1414(2 004) and Li et al., Nat. Biotech. 24:210-215 (2006). Illuminate.

[0125] Suitable host cells for the expression of glycosylated antibodies include multicellular organisms (invertebrates and Examples of invertebrate cells include plant cells and insect cells. In particular, for transfection of fall armyworm cells, binding to insect cells is A number of baculovirus strains have been identified that can be used in various cases.

[0126] Vertebrate cells may also be used as hosts, e.g., cells adapted to grow in suspension. Other examples of useful mammalian host cell lines include: The SV40-transformed monkey kidney CV1 strain (COS-7) and the human embryonic kidney strain ( For example, the methods described in Graham et al., J. GenVirol. 36:59 (1977) 293 or 293 cells), baby hamster kidney cells (BHK), and mouse cells Avian cells (e.g., Mather, Biol. Reprod. 23:243-251 (1 TM4 cells as described in (980), monkey kidney cells (CV1), and African midge cells. Rizal kidney cells (VERO-76), human cervical cancer cells (HELA), and canine kidney cells (MDCK), buffalo rat hepatocytes (BRL 3A), and human lung cells (W138 ), human hepatocytes (Hep G2), mouse mammary tumor (MMT 060562), e.g. For example, Mather et al., Annals NYAcad.Sci.383:44-68 (1982), TRI cells, MRC5 cells, and FS4 cells. Other useful mammalian host cell lines include Chinese hamster ovary (CHO) cells, including DHFR-CHO cells. Human ovarian (CHO) cells (Urlaub et al., Proc. Natl. Acad. Sci. U SA 77:4216(1980)), and myeloma cells such as Y0, NS0, and Sp2 / 0. For a review of specific mammalian host cell lines suitable for antibody production, see, e.g., Yazaki and Wu, Methods in Molecular Biolog y, Vol.248 (edited by BKCLo, Humana Press, Totowa, NJ), pp. 255-268 (2003).

[0127] Pharmaceutical compositions and therapeutic uses of anti-NAMTP antibodies Methods for treating inflammation-related conditions are also provided. Some embodiments involve the use of anti-NAMP antibodies. Administration of T antibodies or antigen-binding fragments thereof dose-dependently reduces inflammation, injury, and BA This includes reducing the amount of protein in the L fluid and / or BAL PMNs. Embodiments include eNAMPT-induced lung injury, lipopolysaccharide-induced lung injury, and / or ventilator-induced lung injury. This includes reducing lung injury caused by NAMPT. Possible diseases include, but are not limited to, pulmonary fibrosis (IPF), pulmonary hypertension, and acute lung injury (ALI). ), acute respiratory distress syndrome (ARDS), ventilator-induced lung injury (VILI), ARDS / VILI-induced ALI, trauma-induced acute lung injury (TIALI), brain injury (traumatic brain injury) Inflammatory diseases such as radiation-induced lung injury (RILI), prostate cancer, lung cancer, , inflammation-related cancers, and pregnant women with chorioamnionitis (e.g., preterm labor and maternal / neonatal complications) (subjects at risk for neonatal complications), nonalcoholic steatohepatitis (NASH), and liver fibrosis cardiac ischemia, and cardiac fibrosis.

[0128] an anti-NAMPT antibody or an antigen-binding fragment thereof, and optionally one or more carriers; Also provided are compositions (e.g., pharmaceutical compositions) further comprising a diluent, excipient, or other additive. Some embodiments include a polynucleotide or vector, optionally comprising one or more and compositions (e.g., pharmaceutical compositions) further comprising a carrier, diluent, excipient, or other additive of .

[0129] Anti-NAMPT antibodies or antigen-binding fragments thereof are produced and administered to a subject in need thereof. For example, NAMPT-related proteins, which are well known or can be easily determined by the skilled artisan. A range of acute and chronic inflammatory disorders (e.g., ARDS, VILI, and trauma-induced inflammatory lung injury) Also provided are methods for reducing and / or treating symptoms, morbidity, or mortality associated with anti-wounds. The NAMPT antibody or antigen-binding fragment thereof can be administered intravenously, intraarterially, intraperitoneally, intrathoracically, or intravenously. Intrathecal, intratracheal, topical, subcutaneous, mucosal, intrapericardial, oral, topical, by inhalation, by injection, By injection, by continuous infusion, by direct local perfusion of the target cells, via a catheter and administering the compound to a subject via aerosol, via nebulizer, and / or via lavage. In some embodiments, the anti-NAMPT antibody or antigen-binding fragment thereof The composition containing the compound is applied directly to a tissue or organ that is inflamed or showing signs of inflammation. Typically, suitable pharmaceutical compositions include, but are not limited to, buffers (e.g., acetate, phosphate, salt or citrate buffers), surfactants (e.g., polysorbates), stabilizers (e.g., , human albumin), etc.

[0130] Certain pharmaceutical compositions provided herein may be administered in the form of, for example, capsules, tablets, aqueous suspensions, or the like. The composition may be administered orally in any acceptable dosage form, including a liquid or solution. These compositions may be administered by nasal aerosol or inhalation. or saline solution with other suitable preservatives and absorption enhancers to increase bioavailability The composition may be prepared as a solution in, and / or other conventional solubilizing or dispersing agents.

[0131] Anti-NAMPT antibodies or their derivatives that can be combined with carrier materials to form a single dosage form. The amount of antigen-binding fragment will vary depending on the subject being treated and the particular mode of administration. The composition may be administered as a single dose, multiple doses, or in an infusion over an established period of time. The dosage regimen also provides the optimum desired response (e.g., a therapeutic or prophylactic response). In one example, the antibody may be administered at a dose of about 0.1 mg / kg, approximately weekly to monthly. It may be administered / administerable intravenously to a patient in an amount of up to about 20 mg / kg.

[0132] In some embodiments, the intended goal is to prevent or treat an inflammatory disease or disorder. The goal is to provide an effective amount that will provide some prevention or treatment of an inflammatory disease or disorder. In some embodiments, this goal is achieved by specific diseases or illnesses, including but not limited to VILI, ALI, or ARDS and preventing or alleviating symptoms and / or cellular processes associated with, such symptoms include, The inflammatory disease or condition may be vascular permeability or elevated BAL protein secretion. Inflammation refers to a disease or illness characterized by inflammation in the heart, lungs, kidneys, liver, bone marrow, pancreas, brain, and skin. skin, bones, veins, arteries, corneas, ears, eyes, nasopharyngeal tissues, stomach, joints, cartilage, vascular tissues or cells, Blood, small intestine, large intestine, larynx, brain, spinal cord, smooth muscle, nerves, skeletal muscle, breast, ovaries, testes, uterus, and It may affect tissues or organs such as platelets, bone marrow cells, and the umbilical cord. , red blood cells, lymphocytes, adipocytes, fibroblasts, epithelial cells, endothelial cells, smooth muscle cells, skeletal muscle cells, endocrine cells, glial cells, neurons, secretory cells, barrier function cells, contractile cells, Absorbent cells, mucosal cells, marginal cells (corneal derived), stem cells (totipotent, pluripotent or multipotent) The cells may include one or more cell types, such as unfertilized or fertilized egg cells or sperm.

[0133] In some embodiments, one or more agents may be administered to a patient, and may be administered simultaneously. Exemplary agents include: are azathioprine, bortezomib, carfilzomib, cyclophosphamide, and dexamethasone. Tazone, doxorubicin, lenalidomide, melphalan, pomalidomide, prednisolone , thalidomide, and vincristine. Exemplary drugs also include antibiotics. Exemplary agents may also be administered / administerable as part of a treatment regimen. stomach.

[0134] The present disclosure further provides pharmaceutical packs and kits comprising one or more containers, the containers comprising the pharmaceutical composition of the present disclosure. and one or more doses of an anti-N antibody containing a composition that can be used to practice the methods described herein. In certain embodiments, the kit may comprise an AMPT antibody or antigen-binding fragment. The composition contains at least one purified anti-NAMPT antibody or antigen-binding fragment thereof. Those skilled in the art will appreciate that the disclosed anti-NAMPT antibodies are compatible with established kinases well known to those of skill in the art. It will be readily apparent that the above-described method can be readily incorporated into one of the various standard formats.

[0135] In certain aspects, the present disclosure provides a method for treating NAMPT-related acute and chronic myocardial infarction in a subject in need thereof. Preventing, reducing and / or reversing the pathophysiological processes that lead to the development and progression of chronic inflammation The present invention provides a prophylactic and / or therapeutic method for treating NAMPT, the method comprising administering an effective amount of an anti-NAMPT antibody. or antigen-binding fragments thereof (including the compositions and pharmaceutical compositions provided herein). As used herein, "NAMPT-associated acute "and chronic inflammatory disorders or diseases" refers to inflammation involving NAMPT, e.g., inflammatory cytokines. Expression and / or function of NAMPT (e.g., eNAMPT), including downstream signaling molecules such as ATP In certain embodiments, the present invention includes any inflammatory disease or biological process that results in increased activity of the inflammatory cytokines. In certain embodiments, the composition comprises a neutralizing humanized anti-NAMPT antibody. The subject is a mammal, including, but not limited to, a human. and corresponding uses to slow, stop, or reverse one or more symptoms associated with the disorder. This can improve the quality of life of the subject and / or extend the lifespan of the patient.

[0136] In some embodiments, an anti-NAMPT antibody or a fragment thereof described herein Symptoms of acute or chronic inflammatory disease or disorder (e.g., inflammatory lung disease or disorder) or have an acute or chronic inflammatory disease or disorder (e.g., an inflammatory lung condition or disorder). It is used to treat patients who are at risk of developing the disease.

[0137] Subjects who should benefit from the methods provided by the present disclosure include critically ill subjects and Critically ill subjects with respiratory failure and critically ill subjects exposed to infection, trauma and / or sepsis; Subjects with radiation exposure, subjects diagnosed with pulmonary fibrosis (IPF), and subjects with pulmonary hypertension Subjects with acute respiratory distress syndrome (ARDS) and subjects with ventilator-induced lung injury Subjects with respiratory failure or VILI and / or ARDS Intensive care unit (ICU) subjects at risk for ALI induced by VILI and pancreatitis Subjects with smoke inhalation injury, blast injury, and / or trauma-induced acute lung injury (TIALI) subjects with traumatic brain injury; subjects with hemorrhagic shock and resuscitation; Subjects with radiation-induced lung injury (including cancer treatments associated with radiation-induced lung injury) and villous Pregnant subjects with amnionitis (e.g., preterm labor and maternal / neonatal complications, or preterm labor) Those at risk for complications in their children and those with primary and metastatic cancer. and the risk of developing any of the diseases disclosed herein. Subjects with

[0138] In certain aspects, the present disclosure provides a method for treating a lung disease or disorder, the method comprising administering to a patient suffering from or at risk of an inflammatory lung disease or disorder. The present invention provides a method for treating a patient suffering from a cancer, the method comprising administering an effective amount of an anti-NAMPT antibody or its The antigen-binding fragments (including the compositions and pharmaceutical compositions provided herein) can be administered to patients. This administration may be based on the patient's symptoms, medical history, or the results of one or more tests. In some cases, patients may have anti-NAMPT antibodies or antigen-binding When administered, the fragments (including the compositions and pharmaceutical compositions provided herein) In some cases, the patient has already been diagnosed with a pulmonary inflammatory disease or disorder. Not diagnosed with an inflammatory disease or disorder, but at risk for such a disease or disorder. Such patients include those who are or will be placed on a ventilator, those with pneumonia, patients who have experienced physical trauma, severe bleeding, inhaled vomit, or chemicals These include patients who have inhaled chemicals, who smoke heavily, and / or who drink heavily. For example, the patient may be placed on a ventilator or , 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 , 22, 23, 24, 30, 36, 42, 48, 54, 60, 66, 72, 78, 84, Within 90, 96, 102, 108, 114, or 120 hours (or any derivable period therein) range) and / or 1, 2, 3, 4, 5, 6, or 7 days (or any derivable period therein) The patient was placed on a ventilator (range of

[0139] In certain aspects, the present disclosure provides methods for treating acute lung injury (ALI), ventilator-induced lung injury (VLI), and suffer from or are at risk of ILI or Acute Respiratory Distress Syndrome (ARDS) The present invention provides a method for treating a patient suffering from NAMPT, the method comprising administering an effective amount of an anti-NAMPT antibody or its antigen binding molecule to a patient suffering from NAMPT. The conjugated fragments (including the compositions and pharmaceutical compositions provided herein) are administered to a patient. Patients at risk include those with sepsis or symptoms of sepsis, pulmonary Patients with symptoms of ulcerative colitis or pneumonia, patients with severe bleeding due to physical injuries, patients with severe chest or head injuries Patients with wounds, patients who have inhaled harmful fumes or smoke, patients who have inhaled vomit, or patients who have been exposed to multiple or multiple Patients who have received large amounts of blood transfusions, patients with fractured long bones (such as the femur), patients who have nearly drowned, and patients who have had anticoagulants. Patients who have had adverse reactions to cancer drugs or other drugs, who have taken an overdose of drugs, or who have pancreatitis patients, heavy smokers, heavy drinkers, patients with inflammatory bowel disease, rheumatoid arthritis Patients with hives, colon cancer, and obesity-related insulin resistance or any combination thereof.

[0140] In certain aspects, the present disclosure provides a method for preventing ventilator-induced lung injury (VILI) in a patient. The present invention provides a method for administering an effective amount of an anti-NAMPT antibody or an antigen-binding fragment thereof to a subject. (including the compositions and pharmaceutical compositions provided herein). In certain embodiments, the anti-NAMPT antibody comprises a neutralizing antibody. In certain embodiments, administration occurs before the patient is placed on a ventilator. This is done after the patient is placed on a ventilator.

[0141] In certain aspects, the present disclosure provides a method for treating NAMPT-associated acute and / or chronic inflammatory disorders. Reduce the level of or one or more sites in subjects at risk of or suffering from The present invention provides a method for reducing the levels of cytokines (e.g., IL-6, TNF-α, IL-1β, IL-8). To provide.

[0142] In certain aspects, the present disclosure provides a method for treating NAMPT-associated acute and / or chronic inflammatory disorders. and methods for reducing eNAMPT levels in subjects at risk of or suffering from cancer. do.

[0143] In some embodiments, the anti-NAMPT antibodies disclosed herein are useful in treating cancer. In some embodiments, the cancer may be used to treat prostate cancer (PCa). In certain embodiments, the present disclosure provides a therapeutically effective amount of an anti-N by administering an AMPT antibody or antigen-binding fragment thereof to a subject in need thereof Methods for treating PCa in a subject are provided. In some embodiments, the method comprises administering to a subject having prostate cancer a therapeutically effective amount of the ... The subject has recurrent, aggressive, or metastatic PCa. In the method, the subject has aggressive PCa that is resistant to androgen deprivation therapy (ADT). .

[0144] Alternatively, in some cases, the anti-NAMPT antibody or antigen-binding fragment may be administered as a It can be administered to a subject in combination with DT. ADT can be administered to the subject before, simultaneously with, or after administration of the menthol.

[0145] It can be administered to a subject in combination with an anti-NAMPT antibody or antigen-binding fragment. The ADTs used are luteinizing hormone-releasing hormone (LHRH) agonists, LHRH antagonists, and CY may include one or more of a P17 inhibitor, an antiandrogen and / or an androgen suppressant In certain embodiments, the LHRH agonist is leuprorelin (e.g., LUPRO N (registered trademark), ELIGARD (registered trademark), etc.), goserelin (e.g., ZOLADEX (registered trademark), triptorelin (e.g., TRELSTAR®) and / or histamine The LHRH antagonist may be threlin (e.g., VANTAS®), and the LHRH antagonist may be degassing. The CYP17 inhibitor may be a CYP17 inhibitor such as FIRMAGON®. , abiraterone (e.g., ZYTIGA®), and the antiandrogen , flutamide (e.g., EULEXIN®), bicalutamide (e.g., CASO DEX®), nilutamide (e.g., NILANDRON®), Enzal thamide (e.g., XTANDI®) and / or apalutamide (e.g., ERLE®) ADA®), and / or the androgen suppressant may be an estrogen and and / or ketoconazole (e.g., NIZORAL®).

[0146] Some embodiments include one or more coronavirus disease 2019 (COVID-19) COVID-19 is a novel coronavirus caused by SARS-CoV-2 (SARS-CoV-2) and is a novel coronavirus caused by SARS-CoV-2. SARS-CoV-2 is a severe acute respiratory syndrome caused by SARS-CoV-2. oV-2 has diameters ranging from 60 nm to 140 nm, with characteristic spiking in the range of 9 nm to 12 nm. Coronaviruses are genetically modified and It can adapt to and infect new hosts through mutation. SARS-CoV-2 infection It may be asymptomatic or may cause a wide range of symptoms. Symptoms include fever, cough, shortness of breath, weakness, fatigue, nausea, vomiting, and changes in taste and smell. Adverse consequences include disseminated intravascular coagulation, inflamed lung tissue and pulmonary endothelial cells, Deep vein thrombosis, pulmonary embolism, thrombotic arterial complications (e.g., limb ischemia, ischemic stroke, myocardial infarction) For SARS-CoV-2 infection, please see Wiersinga et al., "Pathophysiol ogy, Transmission, Diagnosis, and Treatment of Coronavirus Disease 2019(COVID-2019) :A Review,” JAMA, doi:10.1001 / jama.2020.12 839 (published online July 10, 2020). In some embodiments, the present disclosure provides a method for treating NAMPT comprising administering an effective amount of an anti-NAMPT antibody or its antigen-binding fragment to a subject. The fragment may be administered to a subject with COVID-19 (e.g., a subject with COVD-19) by administering the fragment to the subject. Subjects diagnosed with COVID-19 and / or exhibiting one or more symptoms of COVID-19 The present invention provides a method for treating rheumatoid arthritis. [Example]

[0147] The following examples are included for illustrative purposes only and are not intended to be limiting. There is no.

[0148] Example 1. Generation of anti-human NAMPT monoclonal antibodies Three mice were transfected with recombinant extracellular human NAMPT (hNAMPT, MBL International) Neutralizing anti-NAMPT mouse antibodies were raised by immunizing mice with IgG (Chemical). Mouse 4C6 and mouse 589 jointly generated 52 parental clone anti-NAMPT antibodies. The binding of mouse antibodies to NAMPT was assessed by ELISA. In vitro neutralization was investigated by analyzing the effect of antibodies on induced NFκB phosphorylation. The anti-hNAMPT antibodies AL-303 and AL-310 were derived from recombinant hNAMPT. Binds to NAMPT and hNAMPT in lysates from human pulmonary artery endothelial cells (HPAEC) These antibodies were selected because of their ability to Both were able to inhibit hNAMPT-induced NFκB phosphorylation. , which could cross-react with recombinant mouse NAMPT (mNAMPT), but not with AL-31 0 was not. Furthermore, as described in Examples 4 and 5 below, these antibodies The body was subjected to in vivo testing.

[0149] AL-303 The mouse anti-hNAMPT antibody AL-303 has the amino acid sequence shown in SEQ ID NO: 54. and a light chain variable region having the amino acid sequence shown in SEQ ID NO: 55. The heavy chain variable region of AL-303 has the amino acid sequence shown in SEQ ID NO:3. a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 4, a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 5, and a CDR3 domain having the sequence shown in SEQ ID NO: 6. The light chain of AL-303 contains a CDR3 domain having the amino acid sequence shown in column number 5. The variable region comprises a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 6, a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 7, and a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 8. It comprises a CDR3 domain having the sequence

[0150] AL-310 The mouse anti-NAMPT antibody AL-310 has the amino acid sequence shown in SEQ ID NO: 56. and a light chain variable region having the amino acid sequence shown in SEQ ID NO: 57. The heavy chain variable region of AL-310 is C 1 H 2 H 3 H 4 H 5 H 6 H 7 H 8 H 9 H 10 H 11 H 12 H 13 H 14 H 15 H 16 H 17 H 18 H 19 H 10 ...1 19 H 12 H 13 H 14 H 15 H 1 a DR1 domain, a CDR2 domain having the amino acid sequence set forth in SEQ ID NO: 20, and AL-310 contains a CDR3 domain having the amino acid sequence set forth in SEQ ID NO: 21. The light chain variable region comprises a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 6, a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 7, a CDR3 domain having the amino acid sequence shown in SEQ ID NO: 8, a CDR4 domain having the amino acid sequence shown in SEQ ID NO: 9, a CDR5 domain having the amino acid sequence shown in SEQ ID NO: 10, a CDR6 domain having the amino acid sequence shown in SEQ ID NO: 11 a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 7, and an amino acid sequence shown in SEQ ID NO: 22; It contains a CDR3 domain having an amino acid sequence.

[0151] The amino acid sequences of the heavy chain variable region and light chain variable region of AL-303 and AL-310, and The CDRs are shown in Table 1.

[0152] [Table 1] [Table 2]

[0153] Example 2. Detection of human NAMPT and mouse NAMPT using mouse anti-NAMPT antibody Mouse anti-NAMPT antibodies AL-303 and AL-310 To assess the immunoreactivity of NAMPT, we performed Western blot analysis on these cells. The detection of hNAMPT and mNAMPT by antibodies was tested. Immunoreactivity of 10 hNAMPT was investigated using recombinant hNAMPT and HPAEC cell lysates. The ability of these antibodies to detect hNAMPT in lysates was also assessed. The immunoreactivity of AL-303 and AL-310 with mNAMPT was investigated using recombinant mNA MPT, mNAMPT in lysates from spontaneously breathing mice ("SB") exposed to LPS mNAMPT in lysates from injured mouse lungs and mouse ventilator-induced lung injury (VILI) ) to evaluate the ability of these antibodies to detect mNAMPT in lysates from the model. Therefore, further testing was carried out.

[0154] The corresponding Western blot analysis results are provided in Figure 1. As shown, AL-303 and AL-310 (compared to other antibodies) strongly immunoreact with hNAMPT. In comparison, the immunoreactivity of these antibodies with mNAMPT was substantially weaker. Of the five mouse antibodies tested, AL-303 and AL-304 (same as AL-303) Only the α- and α-glucan-containing antibody (containing the α- and α-glucan-containing sequence) immunoreacted with recombinant mNAMPT, whereas all antibodies reacted with recombinant hNAMPT. It reacted with AMPT and hNAMPT in HPAEC cells.

[0155] Example 3. Effect of mouse anti-NAMPT antibody on hNAMPT-induced NFκB phosphorylation Mouse anti-NAMPT antibody AL-303 for hNAMPT-induced NFκB phosphorylation Regarding the effects of AL-304, AL-305, AL-309 and AL-310, Phospho-NFκB (p-NFκB) in cells exposed to hNAMPT in the absence or presence of antibodies NFκB) expression was assessed by Western blot analysis.

[0156] Recombinant hNAMPT (1 μg / ml) was administered in the presence of vehicle, 100 μg / ml of anti-NAMPT Polyclonal (pAb) or 100 μg / ml mouse anti-NAMPT antibody (AL-30 3, AL-304, AL-305, AL-309 or AL-310) for 30 minutes. HPAEC cells were stimulated by exposure to the hNAMPT mixture for 1 hour. Vehicle, 100 μg / ml anti-NAMPT pAb, or 100 μg / ml mouse anti-NA MPT antibody (AL-303, AL-304, AL-305, AL-309 or AL-31 Unstimulated cells ("Unstim") exposed to IFN-γ alone (Fv. 0) were used as a negative control. HPAEC cells exposed to TNF-α were used as a positive control. The expression of p-NFκB in lysates from MPT-stimulated and TNF-α-stimulated cells was measured using pN The results were evaluated by Western blot analysis using an FκB-specific antibody. The results of the RT analysis are provided in Figure 2.

[0157] As shown in Figure 2, hNAMPT inhibited NFκB activation in hNAMPT-stimulated cells. This induces phosphorylation of AL-303, AL-304, AL-305, AL-309 or was substantially attenuated in the presence of AL-310. All five antibodies inhibited hNAMPT-induced NF-κB. Although AL-303 and AL-310 effectively reduced κB phosphorylation, the most substantial effect was observed in AL-304, AL-304, AL-305 and AL-309 were observed in the first This anti-hNAMPT antibody was also obtained through mouse screening.

[0158] Example 4. In vitro study of AL-303, AL-304, and AL-305 in a lung injury model exam Mouse anti-NAMPT antibodies AL-303, AL-304, and AL-305 treat lung injury The ability to do so was tested in vivo using a mouse model.

[0159] As shown in Figure 3, 50 μg, 100 μg, or 200 μg of mouse anti-NAMPT Antibodies, AL-303, AL-304, or AL-305, or vehicle alone were administered to C57 / B6 Mice were intravenously injected with 40 μg / ml of anti-NAMPT antibody. One hour after injection, the mice were treated with 40 μg / ml of anti-NAMPT antibody. They were exposed to intratracheal hNAMPT (Peprotech). Eight hours after hNAMPT exposure, Anti-NAMPT antibody was administered again at the same concentration. Control mice were injected with vehicle via the trachea. Intratracheal hNAMPT was administered to the mice, and the mice were either injected with vehicle and not exposed to intratracheal hNAMPT. All animals were sacrificed 24 hours after hNAMPT exposure, and bronchoalveolar lavage (BAL) was performed. ) by analyzing protein expression and the number of BAL-expressing polymorphonuclear neutrophils (PMNs) , and lung injury was assessed.

[0160] The results of the evaluation are provided in Figure 4. As described in Figure 4, In exposed mice, the expression of BAL proteins (Figure 4A) and the number of BAL-expressing PMNs (Figure 4 B) Mouse anti-NAMPT antibodies AL-303, AL-304, and AL-30 5 effectively reduced hNAMPT-mediated induction of BAL PMN numbers and The hNAMPT-induced increase in protein levels was significantly attenuated by AL-303. Therefore, as shown in Figure 4, AL-303, AL-304 and AL-305 are all , effectively attenuated hNAMPT-induced mouse lung injury, but the most substantial effect was seen with AL-3 03. Therefore, AL-303 was selected for further humanization.

[0161] Example 5. In vivo testing of AL-310 in lung injury The ability of the mouse anti-NAMPT antibody AL-310 to treat lung injury was investigated using a mouse model. and tested in vivo.

[0162] As shown in Figure 5, 10 μg, 25 μg, 50 μg, 100 μg or 200 μg 100 μg of mouse anti-NAMPT antibody AL-310, 100 μg of anti-NAMPT pAb, or C57 / B6 mice were intravenously injected with 1 mg / mL of IL-1 alone. kg of intratracheal LPS. Mice injected with vehicle and not exposed to intratracheal LPS were used as positive controls. Mice that had not been exposed to LPS were used as negative controls. All animals were sacrificed 6 hours after LPS exposure. , Lung injury was assessed by analyzing the expression of BAL protein levels.

[0163] The results of the evaluation are provided in Figure 6. As shown in Figure 6, mice exposed to intratracheal LPS The mouse anti-NAMPT antibody AL-310 induced the expression of BAL protein. It effectively reduced LPS-mediated induction of BAL protein levels, an effect that was significantly greater than that of the positive control. The effect was substantially more pronounced compared to that observed with AL-3. These results indicate that 10 effectively alleviated LPS-induced lung injury in mice. AL-310 was selected for humanization.

[0164] Example 6. Humanization of anti-NAMPT antibody AL-303 Based on the results of the in vitro and in vivo tests described in Examples 2 to 4, mice The anti-NAMPT antibody AL-303 was selected for humanization.

[0165] 1076 Humanized anti-hNAMPT antibody The term "1076" collectively refers to the humanized version of the anti-hNAMPT antibody AL- Specifically, the CDRs of the VH and VL chains of AL-303 are cloned from human heavy and The light chain receptor sequence was grafted.

[0166] Following grafting, de novo synthesis of variable domains or mutagenic oligonucleotide primers and / or by polymerase chain reaction by methods well known in the art. Various framework backmutations were introduced into the heavy and / or light chains of each CDR graft. Various combinations of back mutations and other mutations were constructed against the 1076 humanized antibody. hNMAPT antibodies D-1076, G-1076, K-1076, N-1076, P-10 76, V-1076 and X-1076 were produced.

[0167] The amino acid sequences of the heavy chain variable region and light chain variable region of the 1076 humanized anti-hNMAPT antibody are , as shown in Table 2. Table 3 shows the results of the 10 76 humanized anti-hNMAPT antibodies D-1076, G-1076, K-1076, N-107 6. Alignment of the amino acid sequences of the heavy chain CDRs of P-1076, V-1076, and X-1076 Table 4 provides a summary of the results of the 1-HT100 antibody when compared to the mouse anti-NAMPT antibody AL-303. 076 Humanized anti-hNMAPT antibody D-1076, G-1076, K-1076, N-10 The amino acid sequences of the light chain CDRs of 76, P-1076, V-1076, and X-1076 were compared. Blanks in Tables 3 and 4 indicate residues that are the same as AL-303. is doing.

[0168] D-1076 1076 Humanized anti-hNMAPT antibody D-1076 has the amino acid sequence shown in SEQ ID NO: 1. a heavy chain variable region having the amino acid sequence shown in SEQ ID NO: 2; and a light chain variable region having the amino acid sequence shown in SEQ ID NO: 3. The heavy chain variable region of D-1076 has the amino acid sequence shown in SEQ ID NO:3. a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 4, and a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 5. and a CDR3 domain having the amino acid sequence shown in SEQ ID NO: 5. For the heavy chain, one or more back mutations and other mutations were introduced. The light chain variable region comprises a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 6, a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 7, a CDR3 domain having the amino acid sequence shown in SEQ ID NO: 8, a CDR4 domain having the amino acid sequence shown in SEQ ID NO: 9, a CDR5 domain having the amino acid sequence shown in SEQ ID NO: 10, a CDR6 domain having the amino acid sequence shown in SEQ ID NO: 11 a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 7, and a CDR3 domain having the amino acid sequence shown in SEQ ID NO: 8; It contains a CDR3 domain having the amino acid sequence

[0169] G-1076 The humanized anti-hNMAPT antibody G-1076 has the amino acid sequence shown in SEQ ID NO:9. a heavy chain variable region having the amino acid sequence shown in SEQ ID NO: 10; and a light chain variable region having the amino acid sequence shown in SEQ ID NO: 11. The heavy chain variable region of G-1076 has the amino acid sequence shown in SEQ ID NO:3. a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 4; and a CDR3 domain having the amino acid sequence set forth in SEQ ID NO: 5. For the heavy chain of G-1076, one or more back mutations and other mutations were introduced. The light chain variable region of the present invention comprises a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 11, a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 12, a CDR3 domain having the amino acid sequence shown in SEQ ID NO: 13, a CDR4 domain having the amino acid sequence shown in SEQ ID NO: 14, a CDR5 domain having the amino acid sequence shown in SEQ ID NO: 15, a CDR6 domain having the amino acid sequence shown in a CDR2 domain having the amino acid sequence shown in sequence number 12, and a CDR3 domain having the amino acid sequence shown in sequence number 8; It comprises a CDR3 domain having an amino acid sequence

[0170] K-1076 1076 Humanized anti-hNMAPT antibody K-1076 has the amino acid sequence shown in SEQ ID NO:9. a heavy chain variable region having the amino acid sequence shown in SEQ ID NO: 13; and a light chain variable region having the amino acid sequence shown in SEQ ID NO: 14. The heavy chain variable region of K-1076 has the amino acid sequence shown in SEQ ID NO:3. a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 4; and a CDR3 domain having the amino acid sequence set forth in SEQ ID NO: 5. For the heavy chain of K-1076, one or more back mutations and other mutations were introduced. The light chain variable region of the present invention comprises a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 11, a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 12, a CDR3 domain having the amino acid sequence shown in SEQ ID NO: 13, a CDR4 domain having the amino acid sequence shown in SEQ ID NO: 14, a CDR5 domain having the amino acid sequence shown in SEQ ID NO: 15, a CDR6 domain having the amino acid sequence shown in A CDR2 domain having the amino acid sequence shown in sequence number 14, and a CDR2 domain having the amino acid sequence shown in sequence number 8 It comprises a CDR3 domain having an amino acid sequence

[0171] N-1076 1076 Humanized anti-hNMAPT antibody N-1076 has the amino acid sequence shown in SEQ ID NO: 15 a heavy chain variable region having the amino acid sequence shown in SEQ ID NO: 2; and a light chain variable region having the amino acid sequence shown in SEQ ID NO: 2. The heavy chain variable region of N-1076 has the amino acid sequence shown in SEQ ID NO:3. a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 4; and a CDR3 domain having the amino acid sequence shown in SEQ ID NO: 5. For the heavy chain of N-1076, one or more back mutations and other mutations were introduced. The light chain variable region of a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 7, and an amino acid sequence shown in SEQ ID NO: 8; It contains a CDR3 domain having an amino acid sequence.

[0172] P-1076 1076 Humanized anti-hNMAPT antibody P-1076 has the amino acid sequence shown in SEQ ID NO: 15 a heavy chain variable region having the amino acid sequence shown in SEQ ID NO: 13; and a light chain variable region having the amino acid sequence shown in SEQ ID NO: 14. The heavy chain variable region of P-1076 has the amino acid sequence shown in SEQ ID NO:3. a CDR1 domain having the amino acid sequence shown in SEQ ID NO:4; and a CDR2 domain having the amino acid sequence shown in SEQ ID NO:4. and a CDR3 domain having the amino acid sequence set forth in SEQ ID NO: 5. For the heavy chain of P-107, one or more back mutations and other mutations were introduced. The light chain variable region of SEQ ID NO: 6 comprises a CDR1 domain having the amino acid sequence set forth in SEQ ID NO: 11; a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 14 and a CDR3 domain having the amino acid sequence shown in SEQ ID NO: 8; The CDR3 domain has an amino acid sequence as follows:

[0173] V-1076 1076 Humanized anti-hNMAPT antibody V-1076 has the amino acid sequence shown in SEQ ID NO: 16 a heavy chain variable region having the amino acid sequence shown in SEQ ID NO: 10; and a light chain variable region having the amino acid sequence shown in SEQ ID NO: 11. The heavy chain variable region of V-1076 has the amino acid sequence shown in SEQ ID NO:3. a CDR1 domain having the amino acid sequence shown in SEQ ID NO:4; and a CDR2 domain having the amino acid sequence shown in SEQ ID NO:4. and a CDR3 domain having the amino acid sequence set forth in SEQ ID NO: 5. For the heavy chain of V-107, one or more back mutations and other mutations were introduced. The light chain variable region of SEQ ID NO: 6 comprises a CDR1 domain having the amino acid sequence set forth in SEQ ID NO: 11; a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 12, and a CDR3 domain having the amino acid sequence shown in SEQ ID NO: 8; The CDR3 domain has an amino acid sequence as set forth above.

[0174] X-1076 1076 Humanized anti-hNMAPT antibody X-1076 has the amino acid sequence shown in SEQ ID NO: 16 a heavy chain variable region having the amino acid sequence shown in SEQ ID NO: 2; and a light chain variable region having the amino acid sequence shown in SEQ ID NO: 2. The heavy chain variable region of X-1076 has the amino acid sequence shown in SEQ ID NO:3. a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 4; and a CDR3 domain having the amino acid sequence set forth in SEQ ID NO: 5. One or more back mutations and other mutations were introduced into the heavy chain of X-1076. The light chain variable region of a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 7, and an amino acid sequence shown in SEQ ID NO: 8; It contains a CDR3 domain having an amino acid sequence.

[0175] [Table 3] [Table 4] [Table 5]

[0176] [Table 6] [Table 7]

[0177] [Table 8] [Table 9]

[0178] Example 7. Humanization of anti-NAMPT antibody AL-310 Based on the results of the in vitro and in vivo tests described in Examples 2 to 5, mice The anti-NAMPT antibody AL-310 was selected for humanization.

[0179] "1093" is a set of humanized antibodies derived from the murine anti-NAMPT antibody AL-310. By applying humanization methodology, the CDRs of the VH and VL chains of AL-310 were The sequences were grafted onto different human heavy chain receptor sequences and human light chain receptor sequences.

[0180] The corresponding VH and VL CDRs of AL-310 were grafted onto these receptor sequences. By this, CDR-grafted, humanized and modified VH and VL sequences were generated.

[0181] To generate a humanized antibody with potential framework back mutations, and de novo synthesis or mutagenic oligonucleotide primers for the variable domains. and polymerase chain reaction, or both, by methods well known in the art. Mutations were introduced into the CDR-grafted antibody sequence. For each CDR-graft, the heavy and / or light chains Various combinations of back mutations and other mutations were constructed to generate 1093 humanized anti-hNM APT antibodies FF-1093, II-1093, NN-1093, PP-1093, SS- 1093, UU-1093, XX-1093 and ZZ-1093 were produced.

[0182] The amino acid sequences of the heavy chain variable region and light chain variable region of the 1093 humanized anti-hNMAPT antibody are , as shown in Table 5. Table 6 shows the results of the 10 93 Humanized anti-hNMAPT antibody FF-1093, II-1093, NN-1093, PP -1093, SS-1093, UU-1093, XX-1093 and ZZ-1093 Table 7 provides an alignment of the amino acid sequences of the CDRs of the mouse anti-NAMPT antibodies. When compared with AL-310, 1093 humanized anti-hNMAPT antibody FF-1093, II -1093, NN-1093, PP-1093, SS-1093, UU-1093, XX provides an alignment of the amino acid sequences of the light chain CDRs of -1093 and ZZ-1093. Blank spaces in Tables 6 and 7 indicate residues that are the same as in AL-310.

[0183] FF-1093 1093 Humanized anti-hNMAPT antibody FF-1093 has the amino acid sequence shown in SEQ ID NO: 17. a heavy chain variable region having the amino acid sequence shown in SEQ ID NO: 18; and a light chain variable region having the amino acid sequence shown in SEQ ID NO: 19. The heavy chain variable region of FF-1093 has the amino acid sequence shown in SEQ ID NO: 19. a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 20; domain, and a CDR3 domain having the amino acid sequence shown in SEQ ID NO:21. The heavy chain of FF-1093 contained one or more back mutations and other mutations. The light chain variable region of FF-1093 has a CDR having the amino acid sequence shown in SEQ ID NO:6. 1 domain, a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 7, and a CDR3 domain having the amino acid sequence shown in SEQ ID NO: It contains a CDR3 domain having the amino acid sequence shown in SEQ ID NO:22.

[0184] II-1093 1093 Humanized anti-hNMAPT antibody II-1093 has the amino acid sequence shown in SEQ ID NO:23. a heavy chain variable region having the amino acid sequence shown in SEQ ID NO:24; and a light chain variable region having the amino acid sequence shown in SEQ ID NO:25. The heavy chain variable region of II-1093 has the amino acid sequence shown in SEQ ID NO: 19. a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 20; domain, and a CDR3 domain having the amino acid sequence shown in SEQ ID NO:21. The heavy chain of II-1093 contained one or more backmutations and other mutations. The light chain variable region of II-1093 comprises a CDR having the amino acid sequence shown in SEQ ID NO:6. 1 domain, a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 7, and a CDR3 domain having the amino acid sequence shown in SEQ ID NO: It contains a CDR3 domain having the amino acid sequence shown in SEQ ID NO:22.

[0185] NN-1093 1093 Humanized anti-hNMAPT antibody NN-1093 has the amino acid sequence shown in SEQ ID NO:25. a heavy chain variable region having the amino acid sequence shown in SEQ ID NO:24; and a light chain variable region having the amino acid sequence shown in SEQ ID NO:25. The heavy chain variable region of NN-1093 has the amino acid sequence set forth in SEQ ID NO: 19. a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 20; domain, and a CDR3 domain having the amino acid sequence shown in SEQ ID NO:21. The heavy chain of NN-1093 contained one or more back mutations and other mutations. The light chain variable region of NN-1093 comprises a CDR having the amino acid sequence shown in SEQ ID NO:6. 1 domain, a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 7, and a CDR3 domain having the amino acid sequence shown in SEQ ID NO: It contains a CDR3 domain having the amino acid sequence shown in SEQ ID NO:22.

[0186] PP-1093 1093 Humanized anti-hNMAPT antibody PP-1093 has the amino acid sequence shown in SEQ ID NO:25. a heavy chain variable region having the amino acid sequence shown in SEQ ID NO: 18; and a light chain variable region having the amino acid sequence shown in SEQ ID NO: 19. The heavy chain variable region of PP-1093 has the amino acid sequence shown in SEQ ID NO: 19. a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 20; domain, and a CDR3 domain having the amino acid sequence shown in SEQ ID NO:21. The heavy chain of PP-1093 contained one or more back mutations and other mutations. The light chain variable region of PP-1093 has a CDR having the amino acid sequence shown in SEQ ID NO:6. 1 domain, a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 7, and a CDR3 domain having the amino acid sequence shown in SEQ ID NO: It contains a CDR3 domain having the amino acid sequence shown in SEQ ID NO:22.

[0187] SS-1093 1093 Humanized anti-hNMAPT antibody SS-1093 has the amino acid sequence shown in SEQ ID NO:26. a heavy chain variable region having the amino acid sequence shown in SEQ ID NO:24; and a light chain variable region having the amino acid sequence shown in SEQ ID NO:25. The heavy chain variable region of SS-1093 has the amino acid sequence shown in SEQ ID NO: 19. a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 20; domain, and a CDR3 domain having the amino acid sequence shown in SEQ ID NO:21. The heavy chain of SS-1093 contained one or more back mutations and other mutations. The light chain variable region of SS-1093 comprises a CDR having the amino acid sequence shown in SEQ ID NO:6. 1 domain, a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 7, and a CDR3 domain having the amino acid sequence shown in SEQ ID NO: It contains a CDR3 domain having the amino acid sequence shown in SEQ ID NO:22.

[0188] UU-1093 1093 Humanized anti-hNMAPT antibody UU-1093 has the amino acid sequence shown in SEQ ID NO:26. a heavy chain variable region having the amino acid sequence shown in SEQ ID NO: 18; and a light chain variable region having the amino acid sequence shown in SEQ ID NO: 19. The heavy chain variable region of UU-1093 has the amino acid sequence shown in SEQ ID NO: 19. a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 20; domain, and a CDR3 domain having the amino acid sequence shown in SEQ ID NO:21. The heavy chain of UU-1093 contained one or more back mutations and other mutations. The light chain variable region of UU-1093 has a CDR having the amino acid sequence shown in SEQ ID NO:6. 1 domain, a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 7, and a CDR3 domain having the amino acid sequence shown in SEQ ID NO: It contains a CDR3 domain having the amino acid sequence shown in SEQ ID NO:22.

[0189] XX-1093 1093 Humanized anti-hNMAPT antibody XX-1093 has the amino acid sequence shown in SEQ ID NO:27 a heavy chain variable region having the amino acid sequence shown in SEQ ID NO:24; and a light chain variable region having the amino acid sequence shown in SEQ ID NO:25. The heavy chain variable region of XX-1093 has the amino acid sequence set forth in SEQ ID NO: 19. a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 20; domain, and a CDR3 domain having the amino acid sequence shown in SEQ ID NO:21. The heavy chain of XX-1093 contained one or more back mutations and other mutations. The light chain variable region of XX-1093 has a CDR having the amino acid sequence shown in SEQ ID NO:6. 1 domain, a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 7, and a CDR3 domain having the amino acid sequence shown in SEQ ID NO: It contains a CDR3 domain having the amino acid sequence shown in SEQ ID NO:22.

[0190] ZZ-1093 1093 Humanized anti-hNMAPT antibody ZZ-1093 has the amino acid sequence shown in SEQ ID NO:27. a heavy chain variable region having the amino acid sequence shown in SEQ ID NO: 18; and a light chain variable region having the amino acid sequence shown in SEQ ID NO: 19. The heavy chain variable region of ZZ-1093 has the amino acid sequence shown in SEQ ID NO: 19. a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 20; domain, and a CDR3 domain having the amino acid sequence shown in SEQ ID NO:21. The heavy chain of ZZ-1093 contained one or more back mutations and other mutations. The light chain variable region of ZZ-1093 has a CDR having the amino acid sequence shown in SEQ ID NO:6. 1 domain, a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 7, and a CDR3 domain having the amino acid sequence shown in SEQ ID NO: It contains a CDR3 domain having the amino acid sequence shown in SEQ ID NO:22.

[0191] [Table 10] [Table 11] [Table 12] [Table 13]

[0192] [Table 14] [Table 15]

[0193] [Table 16] [Table 17]

[0194] Example 8. Epitope mapping of humanized anti-hNAMPT antibodies Both linear and conformational epitope mapping were used (performed Humanized anti-NAMPT antibody K-1076 (described in Example 6) and (described in Example 7) The epitope of the humanized anti-NAMPT antibody NN-1093 was mapped.

[0195] A library of peptide-based peptidomimetics was synthesized using Fmoc-based solid-phase peptide synthesis. To generate a library of linear peptide mimetics, we synthesized the amino acid sequence of human NAMPT. The nucleic acid sequence (SEQ ID NO: 60) was split into overlapping fragments in silicon and then placed on a solid support. Amino-functionalized polypropylene supports were synthesized and grafted with a proprietary hydrophilic polymer formulation. followed by dicyclohexylcarbodiimide (DCC) and N-hydroxybenzotriazoline t-Butyloxycarbonyl-hexamethylenediamine (HOBt) BocHMDA) and subsequent Boc using trifluoroacetic acid (TFA) The custom modified JAN was obtained by cleavage of the .GAMMA. group. Amino acid was added by US Liquid Handling Station (Perkin Elmer) The peptides were synthesized on functionalized solid supports.

[0196] Structures using Pepscan's proprietary Chemically Linked Peptides on Scaffolds (CLIPS) technology The CLIPS technology allows the synthesis of peptides with single, double, or triple loop structures. Constructing into loops, sheet-like folds, helix-like folds, and combinations thereof The CLIPS template is attached to a cysteine ​​residue. Cysteine ​​side chains were attached to one or two CLIPS templates, e.g., P2 CL A 0.5 mM solution of IPS (2,6-bis(bromomethyl)pyridine) was dissolved in ammonium bicarbonate. The solution was dissolved in 20 mM sodium chloride (pH 7.8) / acetonitrile (1:3 (v / v)). The CLIPS template was added to the peptide array (3 μl of Two cysteines present in the solid-phase-bound peptides (455-well plate with The peptide array was left in solution for 30–40 min while being completely covered in the solution. The mixture was gently shaken for 60 minutes. Finally, the peptide array was thoroughly washed with excess H2O. , 1% SDS / 0.1% 2,2'-(ethylenedioxy)diene in PBS (pH 7.2) Sonication in disruption buffer containing ethanethiol for 30 min at 70 °C, followed by HO The T3 CLIPS-retained peptide was prepared in the same manner. However, three cysteines were used.

[0197] Binding of the antibodies to each of the synthesized peptides was assessed using a Pepscan-based ELISA. The peptide array was incubated with the primary antibody solution (overnight at 4°C). The peptide array was incubated with a 1 / 1000 dilution of the appropriate detection antibody (antibody peroxidase conjugate) The cells were incubated with HRP-conjugated goat anti-human antibody (Sout Hern Biotech, catalog number 2010-05) was used as the detection antibody. After washing, the peroxidase substrate 2,2'-azino-di-3-ethylbenzthiazoline acetone (ABTS) and 20 μl / ml of 3 percent H2O2 were added. The color development was measured using a charge-coupled device (CCD) camera and an image processing system. did.

[0198] The values ​​obtained from the CCD camera are the same as those obtained from a standard 96-well plate ELISA reader. Similarly, the range was 0–3000 mAU. The results were quantified and compiled into the Peplab database. Occasionally, the wells contain air bubbles, resulting in false positive values, and therefore The cards were manually inspected and any values ​​resulting from air bubbles were scored as 0.

[0199] To verify the quality of the synthesized peptides, a separate set of positive and negative control peptides was run. Control peptides were synthesized in parallel and screened using the commercial antibodies 3C9 and 57.9. Cleaned (Posthumus et al., J Virol, 64:3304-330 9, 1990).

[0200] Data analysis and interpretation were performed using boxplot, linear intensity profile and heatmap analysis. It was carried out.

[0201] Ten different peptides ranging in size from 9 to 30 amino acids were analyzed as described below. The following sets (Set 1 to Set 10) were synthesized.

[0202] Set 1 (LIN15) The peptides of Set 1, also referred to herein as LIN15 peptides, consist of a single residue oncoprotein. It is a 15 amino acid linear peptide derived from the target sequence of NAMPT with a set .

[0203] Set 2 (LIN15.AA) The peptides of Set 2, also referred to herein as LIN15.AA peptides, are It is similar to the LIN1 or LIN15 peptide, but the residues at positions 10 and 11 are replaced by Ala. The natural Ala is replaced by When occurring in the α-terminal position, it was replaced by Gly.

[0204] Set 3 (LIN30) The peptides of Set 3, also referred to herein as LIN30 peptides, contain a single residue on It is a 30 amino acid linear peptide derived from the target sequence of NAMPT with a set .

[0205] Set 4 (LOOP7) The peptides of Set 4, also referred to herein as LOOP7 peptides, consist of 9 amino acids Positions 2-8 contain a 1-residue offset of NAMPT. A 7-mer peptide derived from the target sequence is inserted into Cys to create a loop mimic. Residues were inserted at positions 1 and 9 and attached with mP2 CLIPS. The native Cys was replaced by Cys Replaced by -acm (displayed as "2").

[0206] Set 5 (LOOP15) The peptides of Set 5, also referred to herein as LOOP15 peptides, are 17 amino acids long. It is a restricted peptide of 1000-mer length. Positions 2-16 contain NAM with a 1-residue offset. There is a 15-mer peptide derived from the target sequence of PT (SEQ ID NO: 60). Loop mimic Cys residues were inserted at positions 1 and 17 to create mP2 CLIPS. The native Cys was replaced by Cys-acm (denoted as "2").

[0207] Set 6 (LOOP15.AA) The peptides of Set 6, also referred to herein as LOOP15.AA peptides, are It is similar to the LOOP5 or LOOP15 peptide, but with Ala at positions 10 and 11. The natural Ala is replaced by If it occurs at any position, it is replaced by Gly.

[0208] Set 7 (LOOP25) The peptides of Set 7, also referred to herein as LOOP25 peptides, are 27 amino acids long. It is a restricted peptide of 1000-mer length. Positions 2-26 contain NAM with a 1-residue offset. There is a 25-mer peptide derived from the target sequence of PT (SEQ ID NO: 60). Loop mimic Cys residues were inserted at positions 1 and 27 to create mP2 CLIPS. The native Cys was replaced by Cys-acm (denoted as "2").

[0209] Set 8 (BET) The peptides of Set 8, also referred to herein as BET peptides, are 22 amino acids long. Positions 2-21 contain NA with a one-residue offset. There is a 20-mer peptide derived from the target sequence of MPT (SEQ ID NO: 60). Residues 11 and 12 are replaced by a “PG” motif to induce β-turn formation. To stabilize the mimetic, Cys residues were inserted at positions 1 and 22, resulting in mP2 CLI. The native Cys was replaced by Cys-acm (labeled "2"). It was replaced.

[0210] Set 9 (HEL.CC) The peptides of Set 9, also referred to herein as HEL.CC peptides, are 1 residue long. 22 amino acids from residues of the target sequence (NAMPT, SEQ ID NO: 60) with an offset It is an α-helical peptide mimetic of 100-kJ / s. Cy The native Cys residue was inserted at positions 1 and 5 and attached with mP2 CLIPS. Replaced by s-acm (displayed as "2").

[0211] Set 10 (HEL.IL) The peptides of Set 10, also referred to herein as HEL.IL peptides, consist of one residue 26 amino acids derived from residues of the target sequence (NAMPT, SEQ ID NO: 60) with an offset of It is an α-helical peptide mimetic of 1000-mer length. It can form a helical secondary structure without covalent constraints. To facilitate construction, Leu and Ile residues were inserted into the sequence.

[0212] Screening Details Antibody binding varies depending on the concentration of antibody and the amount and nature of the competitor protein in the ELISA buffer. It also depends on a combination of factors, including pre-coating conditions (peptide pre-incubation with experimental samples). The specific treatment of the chromatin array affected binding. These details are summarized in Table 8. For epscan buffer and preconditioning (SQ), the numbers represent the relative amounts of competing Proteins are shown (combination of horse serum and ovalbumin).

[0213] [Table 18]

[0214] Both antibodies in this study bind to various different regions throughout the NAMPT sequence. This indicates that the epitopes of both antibodies are discontinuous epitopes. This suggests that incubation with the secondary antibody alone did not produce a signal on the array. All signals observed were specific to the antibody Ab-1076-HC2-LC5. In this case, the core sequence derived from the overlapping peptide sequence is present in both monomers within the NAMPT dimer. The LIN30 mimic appears to form a spanning binding interface. The dominant putative junction is surrounded within the structure by additional sequences derived from lower intensity peaks. The tentative core epitopes of the antibodies are shown below. The antibody bound to the dimeric form of NAMPT.

[0215] The epitope candidates identified for both sequences showed a significant amount of repeated sequence similarity. (e.g., 17 SYKVTHYKQYPPNTSKVYSYFEC REKKT 44 ( Sequence number 61) pair 16 2ATNS REQK K 170 (SEQ ID NO: 63), 29 NT SKVY S YFECREKKTENSKLRK 51 (SEQ ID NO: 72) pair 332 FPVTEN SKG Y K 342 (SEQ ID NO: 67), and 216 KGT DTV AGLALIKKYY GTK 23 4 (SEQ ID NO: 75) pairs 316 NPL DTV LKVLEIL GKK 331 (SEQ ID NO: 76) .

[0216] [Table 19]

[0217] Example 9. Characterization of humanized anti-hNAMPT antibodies 1076 humanized anti-hNAMPT antibody described in Example 6 and 1076 humanized anti-hNAMPT antibody described in Example 7 The 093 humanized anti-hNAMPT antibody was subjected to in vitro and in vivo testing to identify the lead humanized anti-h A selection of NAMPT antibodies was tested.

[0218] Effect of humanized anti-hNAMPT antibody on hNAMPT-induced inflammatory signaling 1093 Humanized anti-hNAMPT antibody C against hNAMPT-induced inflammatory signaling C-1093, KK-1093, RR-1093, UU-1093 and XX-1093 The effects were assessed by measuring the N-terminal region of the NAMPT signaling pathway in cells exposed to hNAMPT in the presence or absence of these antibodies. This was assessed by assessing FκB activation.

[0219] Recombinant hNAMPT (1.5 μg / ml) was administered in a 109 ml dose of either vehicle or 100 μg / ml. 3 humanized anti-hNAMPT antibodies (CC-1093, KK-1093, RR-1093, UU Human lung endothelial cells (ECs) were premixed with hNAMP (HN-1093 or XX-1093). The non-human primates were stimulated by exposure to the NAMPT mixture for 1 hour. Stimulator cells served as a negative control ("NC"). hNA premixed with vehicle only Stimulator cells exposed to MPT ("--") served as a positive control. Intracellular NFκB was measured by assessing enzyme activity (NFκB-SecNanoLuc). κB activation was evaluated (n = 3-4 for each mAb). The results are provided in Figure 7A.

[0220] As shown in Figure 7A, hNAMPT inhibited NFκB activity in hNAMPT-stimulated cells. This was induced by humanized anti-hNAMPT antibodies CC-1093, KK-1093, and R was substantially attenuated in the presence of R-1093, UU-1093, or XX-1093. Although all antibodies effectively reduced hNAMPT-induced NFκB activation, the most substantial effect was observed in UU-1093 and XX-1093.

[0221] Effect of humanized anti-hNAMPT antibody on hNAMPT-induced decrease in EC barrier integrity EC electrical resistance reflects the lung EC barrier integrity. Humanized anti-hNAMPT antibody UU-109 against hNAMPT-induced reduction in total body 3, and 1076 humanized anti-hNAMPT antibodies H-1076, P-1076, and N-107 The effects of 6 and D-1076 were examined by exposing hNAMPT to the antibody in the absence or presence of these antibodies. The electrical resistance of the EC was evaluated.

[0222] Recombinant hNAMPT (1.5 μg / ml) was administered in the presence of vehicle alone, anti-NAMPT pAb, or are humanized anti-hNAMPT antibodies (UU-1093, H-1076, P-1076, N-10 The cells were premixed with hNAMPT (D-76 or D-1076). The cells were exposed to the hNAMPT mixture for 1 hour. Human lung ECs were stimulated with hNAMPT or pAb premixed with vehicle alone. Stimulator cells exposed to premixed hNAMPT served as controls. The resistance was assessed as a readout of EC barrier integrity. The results of the study are provided in Figure 7B. do.

[0223] As shown in Figure 7B, exposure to hNAMPT premixed with each humanized anti-hNAMPT antibody was The treated ECs showed a substantial induction of EC barrier activity relative to control cells. hNAMPT-mediated reduction of EC barrier activity was observed with 1076 humanized anti-hNAMPT antibody and 1 This was effectively reversed in the presence of 093 humanized anti-hNAMPT antibody.

[0224] In vivo testing of humanized anti-hNAMPT antibody in lung injury 1076 humanized anti-hNAMPT antibodies (V-1076, N-1076, K-1076 and P-1076) and 1093 humanized anti-hNAMPT antibody (SS-1093, CC-109 The ability of XX-1093 and UU-1093 to treat lung injury was evaluated in mice and rats. The pulmonary injury model was tested in vivo.

[0225] To evaluate the effect of humanized anti-hNAMPT antibody on a mouse model of lung injury, -1076, N-1076, K-1076, P-1076, SS-1093, CC-10 93, XX-1093, or UU-1093 for 8 hours with LPS and the last 4 hours with VILI. The exposed mice were injected intravenously with a dose of 0.4 mg / kg. Mice exposed to hNAMPT and VILI served as controls. BAL protein Lung injury in mice was assessed by analyzing the expression of IL-1 and the number of BAL-expressing cells. Furthermore, H&E staining revealed edema and inflammatory cell infiltration in the lung tissue, which can be used as a readout of lung injury. The results of this "two-hit" model of lung injury are shown in Figures 8A, 8B, and 8D will be provided to.

[0226] As depicted in Figure 8A, compared to control mice, BAL protein levels (Figure 8A The number of BAL-expressing cells (left panel of Figure 8A) and BAL-expressing cells (right panel of Figure 8A) were significantly higher than those of the 1076 humanized anti-histidine antibody. NAMPT was effectively reduced in mice injected with any of the tested antibodies. All 076 humanized anti-hNAMPT antibodies prevented lung damage in this "two-hit" mouse model. Although they effectively reduced scarring, the most substantial effect was observed with P-1076.

[0227] As shown in Figure 8B, BAL protein levels (Figure 8B) were significantly higher in the control mice compared to the control mice. The number of BAL-expressing cells (left panel of Figure 8B) and BAL-expressing cells (right panel of Figure 8B) were significantly higher than those of the 1093 humanized anti-histamine antibody. NAMPT was effectively reduced in mice injected with any of the tested antibodies. All 093 humanized anti-hNAMPT antibodies prevented lung damage in this "two-hit" mouse model. Although both effectively reduced scarring, the most substantial effect was observed with UU-1093. The protective effect of UU-1093 in this two-hit lung injury model, as described in D showed a reduction in inflammatory cell infiltration and pulmonary tissue edema in UU-1093-injected mice. So it was further reflected.

[0228] To evaluate the effect of humanized anti-hNAMPT antibody on a rat model of lung injury, Sprague-Dawley rats were exposed to PS at 40 mg / kg, 80 mg / kg, or 1 60 mg / kg of P-1076 was injected intravenously. Rats injected with vehicle only and not exposed to LPS served as positive controls. Untreated rats served as negative controls. Expression of BAL proteins and number of BAL-expressing cells The results of this rat model of lung injury were evaluated by analyzing the following: is provided in Figure 8C.

[0229] Control mice (vehicle-injected and exposed to LPS) as described in Figure 8C Compared with the BAL protein levels (left panel of Fig. 8C ) and the number of BAL-expressing cells ( The right panel of Figure 8C) shows the results of the humanized anti-hNAMPT antibody P-1076 injected mice. was effectively reduced.

[0230] Therefore, as shown in Figure 8, all humanized anti-hNAMPT antibodies tested , effectively attenuated lung injury in an in vivo lung injury model, but the most substantial effect was seen with P- This was observed in P-1076 and UU-1093. , select as lead humanized anti-hNAMPT antibody, and generate improved anti-hNAMPT antibodies It was further modified to

[0231] Example 10. Modification of humanized anti-hNAMPT antibody P-1076 Based on the results of the in vitro and in vivo tests described in Example 9, anti-NAMP The T antibody P-1076 was further purified to generate an improved 1076 anti-hNAMPT antibody. Selected for modification.

[0232] P-1076-mod humanized anti-hNAMPT antibody P-1076-mod modifies the humanized anti-hNAMPT antibody P-1076 This refers to a humanized anti-hNAMPT antibody produced by

[0233] To generate the improved P-1076-mod anti-hNAMPT antibody, the variable domains De novo synthesis or mutagenic oligonucleotide primers and polymerase chain reaction or By both of these methods, one or more mutations can be introduced into the P-10 These mutations were introduced into 76 sequences. These mutations eliminate oxidation sites, reduce or eliminate deamidation. , removal of potential cleavage or fragmentation sites, removal of potential T cell epitopes, and / or The heavy chain and / or the P-1076 heavy chain were introduced to reduce binding of specific T cell epitopes. constructed various combinations of back mutations and other mutations in the light chain to produce P-1076- mod anti-hNAMPT antibodies P-1076-mod1, P-1076-mod2, P-10 76-mod3, P-1076-mod4, P-1076-mod5, P-1076-m od6, P-1076-mod7, P-1076-mod8, P-1076-mod9, P-1076-mod10 and P-1076-mod11 were generated.

[0234] Heavy and light chain variable regions of modified P-1076-mod anti-hNAMPT antibody The amino acid sequence of is provided in Table 10. In Table 10, the mutated residues are shown in bold. Table 11 shows the efficacy of modified P-1076-mod anti-hN compared to P-1076. AMPT antibodies P-1076-mod1, P-1076-mod2, P-1076-mod 3, P-1076-mod4, P-1076-mod5, P-1076-mod6, P- 1076-mod7, P-1076-mod8, P-1076-mod9, P-1076 Alignment of the amino acid sequences of the heavy chain CDRs of P-1076-mod10 and P-1076-mod11 Table 12 shows the results of the modified P-1076-mod antibody compared to P-1076. hNAMPT antibodies P-1076-mod1, P-1076-mod2, P-1076-m od3, P-1076-mod4, P-1076-mod5, P-1076-mod6, P-1076-mod7, P-1076-mod8, P-1076-mod9, P-10 Alignment of the amino acid sequences of the light chain CDRs of 76-mod10 and P-1076-mod11 Blanks in Tables 11 and 12 indicate residues that are the same as P-1076. A summary of the biophysical properties of the modified P-1076-mod anti-hNAMPT antibody is given below. , provided in Table 13.

[0235] P-1076-mod1 P-1076-mod anti-hNAMPT antibody P-1076-mod1 is identified as SEQ ID NO: 28 and a heavy chain variable region having the amino acid sequence shown in SEQ ID NO: 30. The heavy chain variable region of P-1076-mod1 has the sequence shown in SEQ ID NO: 3, a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 29 a CDR2 domain having the sequence shown in SEQ ID NO: 5, and a CDR having the amino acid sequence shown in SEQ ID NO: 6 The heavy chain of P-1076-mod1 contains a D to E mutation. P-1076 was introduced into VH CDR2 to remove potential cleavage or fragmentation sites. The light chain variable region of 1076-mod1 has the amino acid sequence shown in SEQ ID NO: 11. a DR1 domain, a CDR2 domain having the amino acid sequence set forth in SEQ ID NO: 14, and It contains a CDR3 domain having the amino acid sequence set forth in SEQ ID NO: 8. For the light chain of od1, the L-to-V mutation was added to the P-1076 VL framework. regions to remove potential T cell epitopes.

[0236] P-1076-mod2 P-1076-mod anti-hNAMPT antibody P-1076-mod2 is identified as SEQ ID NO: 28 and a heavy chain variable region having the amino acid sequence shown in SEQ ID NO: 31. The heavy chain variable region of P-1076-mod2 has the sequence shown in SEQ ID NO: 3, a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 29 a CDR2 domain having the sequence shown in SEQ ID NO: 5, and a CDR having the amino acid sequence shown in SEQ ID NO: 6 The heavy chain of P-1076-mod2 contains a D to E mutation. P-1076 was introduced into VH CDR2 to remove potential cleavage or fragmentation sites. The light chain variable region of 1076-mod2 has the amino acid sequence shown in SEQ ID NO: 11. a DR1 domain, a CDR2 domain having the amino acid sequence set forth in SEQ ID NO: 14, and It contains a CDR3 domain having the amino acid sequence set forth in SEQ ID NO: 8. For the light chain of od2, the L to I mutation was added to the P-1076 VL framework. regions to remove potential T cell epitopes.

[0237] P-1076-mod3 P-1076-mod anti-hNAMPT antibody P-1076-mod3 is identified as SEQ ID NO: 28 and a heavy chain variable region having the amino acid sequence shown in SEQ ID NO: 32. The heavy chain variable region of P-1076-mod3 has the sequence shown in SEQ ID NO: 3, a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 29 a CDR2 domain having the sequence shown in SEQ ID NO: 5, and a CDR having the amino acid sequence shown in SEQ ID NO: 6 The heavy chain of P-1076-mod3 contains a D to E mutation. P-1076 was introduced into VH CDR2 to remove potential cleavage or fragmentation sites. The light chain variable region of 1076-mod3 has the amino acid sequence shown in SEQ ID NO: 11. a DR1 domain, a CDR2 domain having the amino acid sequence set forth in SEQ ID NO: 33, and It contains a CDR3 domain having the amino acid sequence set forth in SEQ ID NO: 8. For the od3 light chain, the L-to-V mutation was added to the P-1076 VL framework. region and an L to G mutation was introduced into the P-1076 VL CDR2 framework. It was introduced into the nucleotide region to remove potential T cell epitopes.

[0238] P-1076-mod4 P-1076-mod anti-hNAMPT antibody P-1076-mod4 is identified as SEQ ID NO: 28 and a heavy chain variable region having the amino acid sequence shown in SEQ ID NO: 34. The heavy chain variable region of P-1076-mod4 has the sequence shown in SEQ ID NO: 3, a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 29 a CDR2 domain having the sequence shown in SEQ ID NO: 5, and a CDR having the amino acid sequence shown in SEQ ID NO: 6 The heavy chain of P-1076-mod4 contains a D to E mutation. P-1076 was introduced into VH CDR2 to remove potential cleavage or fragmentation sites. The light chain variable region of 1076-mod4 has the amino acid sequence shown in SEQ ID NO: 11. a DR1 domain, a CDR2 domain having the amino acid sequence set forth in SEQ ID NO: 35, and It contains a CDR3 domain having the amino acid sequence set forth in SEQ ID NO: 8. For the od4 light chain, the L to V mutation was added to the P-1076 VL framework. region and an L to E mutation was introduced into the P-1076 VL CDR2 framework. It was introduced into the nucleotide region to remove potential T cell epitopes.

[0239] P-1076-mod5 P-1076-mod anti-hNAMPT antibody P-1076-mod5 is identified as SEQ ID NO: 28 and a heavy chain variable region having the amino acid sequence shown in SEQ ID NO: 36. The heavy chain variable region of P-1076-mod5 has the sequence shown in SEQ ID NO: 3, a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 29 a CDR2 domain having the sequence shown in SEQ ID NO: 5, and a CDR having the amino acid sequence shown in SEQ ID NO: 6 The heavy chain of P-1076-mod5 contains a D to E mutation. P-1076 was introduced into VH CDR2 to remove potential cleavage or fragmentation sites. The light chain variable region of 1076-mod5 has the amino acid sequence shown in SEQ ID NO: 11. a DR1 domain, a CDR2 domain having the amino acid sequence set forth in SEQ ID NO: 37, and It contains a CDR3 domain having the amino acid sequence set forth in SEQ ID NO: 8. For the od5 light chain, the L to I mutation was added to the P-1076 VL framework. region to remove potential T cell epitopes, and an L to E mutation was introduced into the P- 1076 VL CDR2 framework region to enhance the binding of potential T cell epitopes The cases were removed.

[0240] P-1076-mod6 P-1076-mod anti-hNAMPT antibody P-1076-mod6 is identified as SEQ ID NO: 15 and a heavy chain variable region having the amino acid sequence shown in SEQ ID NO: 30. The heavy chain variable region of P-1076-mod6 has the sequence shown in SEQ ID NO: a CDR1 domain having the amino acid sequence shown in SEQ ID NO:3; a CDR2 domain having the amino acid sequence shown in SEQ ID NO:4; a CDR2 domain having the amino acid sequence shown in SEQ ID NO:5; and a CDR3 domain having the amino acid sequence shown in SEQ ID NO:6. The heavy chain of P-1076-mod6 is identical to the P-1076 VH. The light chain variable region of P-1076-mod6 has the amino acid sequence shown in SEQ ID NO: 11. a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 14; and a CDR3 domain having the amino acid sequence set forth in SEQ ID NO: 8. For the light chain of -mod5, an L to V mutation was made in the framework of P-1076 VL. This was introduced into the target region to remove potential T cell epitopes.

[0241] P-1076-mod7 P-1076-mod anti-hNAMPT antibody P-1076-mod7 is identified as SEQ ID NO: 15 and a heavy chain variable region having the amino acid sequence shown in SEQ ID NO: 31. The heavy chain variable region of P-1076-mod7 has the sequence shown in SEQ ID NO: a CDR1 domain having the amino acid sequence shown in SEQ ID NO:3; a CDR2 domain having the amino acid sequence shown in SEQ ID NO:4; a CDR2 domain having the amino acid sequence shown in SEQ ID NO:5; and a CDR3 domain having the amino acid sequence shown in SEQ ID NO:6. The heavy chain of P-1076-mod7 is identical to the P-1076 VH. The light chain variable region of P-1076-mod7 has the amino acid sequence shown in SEQ ID NO: 11. a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 14; and a CDR3 domain having the amino acid sequence set forth in SEQ ID NO: 8. For the light chain of -mod7, the L to I mutation was added in the framework of P-1076 VL. This was introduced into the target region to remove potential T cell epitopes.

[0242] P-1076-mod8 P-1076-mod anti-hNAMPT antibody P-1076-mod8 is identified as SEQ ID NO: 15 and a heavy chain variable region having the amino acid sequence shown in SEQ ID NO: 32. The heavy chain variable region of P-1076-mod8 has the sequence shown in SEQ ID NO: a CDR1 domain having the amino acid sequence shown in SEQ ID NO:3; a CDR2 domain having the amino acid sequence shown in SEQ ID NO:4; a CDR2 domain having the amino acid sequence shown in SEQ ID NO:5; and a CDR3 domain having the amino acid sequence shown in SEQ ID NO:6. The heavy chain of P-1076-mod8 is identical to the P-1076 VH. The light chain variable region of P-1076-mod8 has the amino acid sequence shown in SEQ ID NO: 11. a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 33; and a CDR3 domain having the amino acid sequence set forth in SEQ ID NO: 8. For the light chain of -mod8, an L to V mutation was made in the framework of P-1076 VL. The L to G mutation was introduced into the P-1076 VL CDR2 frame. The work area was introduced to remove potential T cell epitopes.

[0243] P-1076-mod9 P-1076-mod anti-hNAMPT antibody P-1076-mod9 is identified as SEQ ID NO: 15 and a heavy chain variable region having the amino acid sequence shown in SEQ ID NO: 34. The heavy chain variable region of P-1076-mod9 has the sequence shown in SEQ ID NO: a CDR1 domain having the amino acid sequence shown in SEQ ID NO:3; a CDR2 domain having the amino acid sequence shown in SEQ ID NO:4; a CDR2 domain having the amino acid sequence shown in SEQ ID NO:5; and a CDR3 domain having the amino acid sequence shown in SEQ ID NO:6. The heavy chain of P-1076-mod9 is identical to the P-1076 VH. The light chain variable region of P-1076-mod9 has the amino acid sequence shown in SEQ ID NO: 11. a CDR1 domain having the amino acid sequence set forth in SEQ ID NO: 35; and a CDR3 domain having the amino acid sequence set forth in SEQ ID NO: 8. For the light chain of -mod9, an L to V mutation was made in the framework of P-1076 VL. The L to E mutation was introduced into the P-1076 VL CDR2 frame. The work area was introduced to remove potential T cell epitopes.

[0244] P-1076-mod10 P-1076-mod anti-hNAMPT antibody P-1076-mod10 is SEQ ID NO: 15 and a heavy chain variable region having the amino acid sequence shown in SEQ ID NO: 36. and a light chain variable region having the sequence a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 3, an amino acid sequence shown in SEQ ID NO: 4, a CDR2 domain having the amino acid sequence shown in SEQ ID NO:5; The heavy chain of P-1076-mod10 contains the R3 domain. The light chain variable region of P-1076-mod10 has the amino acid sequence shown in SEQ ID NO:11. a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 37; a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 38; P- and CDR3 domains having the amino acid sequence set forth in SEQ ID NO: 8. For the light chain of 1076-mod10, the L to I mutation was introduced into the framework regions to remove potential T cell epitopes, and Mutations were introduced into P-1076 VL CDR2 to block potential T cell epitope binding. Removed.

[0245] P-1076-mod11 The modified P-1076-mod anti-hNAMPT antibody P-1076-mod11 was synthesized using the A heavy chain variable region having the amino acid sequence shown in SEQ ID NO: 28 and a heavy chain variable region having the amino acid sequence shown in SEQ ID NO: 13 and a heavy chain variable region of P-1076-mod11. The region comprises a CDR1 domain having the amino acid sequence shown in SEQ ID NO:3, a CDR2 domain having the amino acid sequence shown in SEQ ID NO:29, and a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 5. The heavy chain of P-1076-mod11 contains a CDR3 domain having the sequence D to A mutation to E was introduced into P-1076 VH CDR2 to remove potential cleavage or fragmentation sites. The light chain variable region of P-1076-mod11 is the amino acid sequence shown in SEQ ID NO:11. a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 14; R2 domain, and a CDR3 domain having the amino acid sequence shown in SEQ ID NO:8 The light chain of P-1076-mod11 is identical to P-1076 VL.

[0246] [Table 20] [Table 21] [Table 22] [Table 23] [Table 24] [Table 25] [Table 26] *In Table 10, the mutated residues are shown in bold.

[0247] [Table 27] [Table 28]

[0248] [Table 29] [Table 30]

[0249] [Table 31]

[0250] Example 11. Modification of humanized anti-hNAMPT antibody UU-1093 Based on the results of the in vitro and in vivo tests described in Example 9, anti-NAMP The antibody UU-1093 was used for further analysis to generate modified 1093 anti-hNAMPT antibodies. were selected for modification.

[0251] UU-1093-mod humanized anti-hNAMPT antibody UU-1093-mod is a modified humanized anti-hNAMPT antibody, UU-1093. The term "humanized anti-hNAMPT antibody" refers to a humanized anti-hNAMPT antibody produced by

[0252] To generate the improved UU-1093-mod anti-hNAMPT antibody, De novo synthesis or mutagenic oligonucleotide primers and polymerase chain reaction or or both, by methods well known in the art, to introduce one or more mutations into the UU- These mutations were introduced into the 1093 amino acid sequence. reduction or elimination, removal of potential cleavage or fragmentation sites, removal of potential T cell epitopes, and / or introduced to reduce binding of potential T cell epitopes. UU-1093 constructing various combinations of back mutations and other mutations in the heavy and / or light chains of Modified UU-1093-mod anti-hNAMPT antibody UU-1093-mod1, UU -1093-mod2, UU-1093-mod3, UU-1093-mod4, UU- 1093-mod5, UU-1093-mod6, UU-1093-mod7, UU-1 093-mod8, UU-1093-mod9, UU-1093-mod10, UU-1 093-mod11, UU-1093-mod12, UU-1093-mod13, UU -1093-mod14, UU-1093-mod15, UU-1093-mod16, UU-1093-mod17, UU-1093-mod18 and UU-1093-mod 19 was generated.

[0253] Heavy and light chain variable regions of modified P-1076-mod anti-hNAMPT antibody The amino acid sequence of is provided in Table 14. In Table 14, the mutated residues are shown in bold. Table 15 shows the effect of modified UU-1093-mod antibody compared to UU-1093. hNAMPT antibodies UU-1093-mod1, UU-1093-mod2, UU-109 3-mod3, UU-1093-mod4, UU-1093-mod5, UU-1093 -mod6, UU-1093-mod7, UU-1093-mod8, UU-1093- mod9, UU-1093-mod10, UU-1093-mod11, UU-1093 -mod12, UU-1093-mod13, UU-1093-mod14, UU-10 93-mod15, UU-1093-mod16, UU-1093-mod17, UU- Amino acid sequences of the heavy chain CDRs of 1093-mod18 and UU-1093-mod19 Table 16 shows the alignment of modified UU-1093 compared to UU-1093. 93-mod anti-hNAMPT antibody UU-1093-mod1, UU-1093-mod2 , UU-1093-mod3, UU-1093-mod4, UU-1093-mod5, UU-1093-mod6, UU-1093-mod7, UU-1093-mod8, U U-1093-mod9, UU-1093-mod10, UU-1093-mod11, UU-1093-mod12, UU-1093-mod13, UU-1093-mod1 4, UU-1093-mod15, UU-1093-mod16, UU-1093-mo The light chain CDR sequences of d17, UU-1093-mod18, and UU-1093-mod19 The amino acid sequence alignments are shown. The blanks in Tables 15 and 16 indicate residues UU-1093 and UU-1094. It shows that they are the same.

[0254] UU-1093-mod1 UU-1093-mod anti-hNAMPT antibody UU-1093-mod1 is SEQ ID NO: 3 a heavy chain variable region having the amino acid sequence shown in SEQ ID NO: 8 and a heavy chain variable region having the amino acid sequence shown in SEQ ID NO: 47; The heavy chain variable region of UU-1093-mod1 has the sequence CDR1 domain having the amino acid sequence shown in column number 19, shown in SEQ ID NO: 39 a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 21; The heavy chain of UU-1093-mod1 contains a CDR3 domain that corresponds to the CDR3 domain from D to E. Mutations were introduced into UU-1093 VH CDR2 to eliminate potential cleavage or fragmentation sites. The light chain variable region of UU-1093-mod1 contains the amino acid sequence shown in SEQ ID NO:11. CDR1 domain having the amino acid sequence shown in SEQ ID NO:7, CDR2 domain having the amino acid sequence shown in SEQ ID NO:7 domain, and a CDR3 domain having the amino acid sequence shown in SEQ ID NO:22. For the light chain of UU-1093-mod1, the N to Q mutation was Deamidation was reduced or eliminated by introducing it into VL CDR1.

[0255] UU-1093-mod2 UU-1093-mod anti-hNAMPT antibody UU-1093-mod2 is SEQ ID NO: 4 a heavy chain variable region having the amino acid sequence shown in SEQ ID NO: 47; The heavy chain variable region of UU-1093-mod2 has the sequence CDR1 domain having the amino acid sequence shown in column number 19, shown in SEQ ID NO: 39 a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 41; The heavy chain of UU-1093-mod2 contains a CDR3 domain that is Mutations were introduced into UU-1093 VH CDR2 to eliminate potential cleavage or fragmentation sites. and an S to T mutation was introduced into UU-1093 VH CDR3 to generate the latent The light chain variable region of UU-1093-mod2 was sequenced as SEQ ID NO: 144444. No. 11, a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 7, a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 22; The light chain of UU-1093-mod2 contains the DR3 domain. Mutations were introduced into UU-1093 VL CDR1 to reduce or eliminate deamidation. Ta.

[0256] UU-1093-mod3 UU-1093-mod anti-hNAMPT antibody UU-1093-mod3 is SEQ ID NO: 4 a heavy chain variable region having the amino acid sequence shown in SEQ ID NO:2 and a heavy chain variable region having the amino acid sequence shown in SEQ ID NO:47 The heavy chain variable region of UU-1093-mod3 has the sequence CDR1 domain having the amino acid sequence shown in column number 43, shown in SEQ ID NO: 39 a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 44; The heavy chain of UU-1093-mod3 contains a CDR3 domain that converts M to I. The following mutation was introduced into UU-1093 VH CDR1 to remove the oxidation site, changing M to V The following mutations were introduced into the UU-1093 VH framework region 2 to identify the oxidation site and potential The D to E mutation was introduced into UU-1093 VH CDR2 to remove the important T cell epitope. to remove potential cleavage or fragmentation sites, and an M to V mutation was introduced into UU-109 3 VH framework region 3 to remove the oxidation site and a K to R mutation Mutations introduced into UU-1093 VH CDR3 to reduce binding of potential T cell epitopes The light chain variable region of UU-1093-mod3 has the amino acid sequence shown in SEQ ID NO: 11. a CDR1 domain having the amino acid sequence shown in SEQ ID NO:7; a CDR2 domain having the amino acid sequence shown in SEQ ID NO:8; and a CDR3 domain having the amino acid sequence set forth in SEQ ID NO: 22. For the light chain of -1093-mod3, the N to Q mutation was Deamidation was reduced or eliminated by introducing it into CDR1.

[0257] UU-1093-mod4 UU-1093-mod anti-hNAMPT antibody UU-1093-mod4 is SEQ ID NO: 4 a heavy chain variable region having the amino acid sequence shown in SEQ ID NO:5 and a heavy chain variable region having the amino acid sequence shown in SEQ ID NO:47 The heavy chain variable region of UU-1093-mod4 has the sequence CDR1 domain having the amino acid sequence shown in column number 46, shown in SEQ ID NO: 39 a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 41; The heavy chain of UU-1093-mod4 contains a CDR3 domain that is The mutations and M to I were introduced into UU-1093 VH CDR1 and oxidized. The site was removed and an M to V mutation was introduced into UU-1093 VH framework region 2. The D to E mutation was introduced to remove the oxidation site and potential T cell epitopes. 1093 into VH CDR2 to remove potential cleavage or fragmentation sites, and into UU-1093 VH framework region 3 to remove the oxidation site, Then, an S to T mutation was introduced into the UU-1093 VH CDR3 to induce latent T cell proliferation. The light chain variable region of UU-1093-mod4 was sequenced as SEQ ID NO: 11. a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 7; and a CDR3 domain having the amino acid sequence shown in SEQ ID NO: 22. For the light chain of UU-1093-mod4, the N to Q mutation was U-1093 was introduced into VL CDR1 to reduce or eliminate deamidation.

[0258] UU-1093-mod5 UU-1093-mod anti-hNAMPT antibody UU-1093-mod5 is SEQ ID NO: 3 a heavy chain variable region having the amino acid sequence shown in SEQ ID NO: 8 and a heavy chain variable region having the amino acid sequence shown in SEQ ID NO: 48 The heavy chain variable region of UU-1093-mod5 has the sequence CDR1 domain having the amino acid sequence shown in column number 19, shown in SEQ ID NO: 39 a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 21; The heavy chain of UU-1093-mod5 contains a CDR3 domain that Mutations were introduced into UU-1093 VH CDR2 to eliminate potential cleavage or fragmentation sites. The light chain variable region of UU-1093-mod5 contains the amino acid sequence shown in SEQ ID NO:49. CDR1 domain having the amino acid sequence shown in SEQ ID NO: 50 2 domain, and a CDR3 domain having the amino acid sequence set forth in SEQ ID NO: 22. For the light chain of UU-1093-mod5, a G to A mutation was introduced. Introducing an I to V mutation into VL CDR1 to reduce or eliminate deamidation was introduced into UU-1093 VL framework region 2 to enhance the binding of potential T cell epitopes. and introducing an M to V mutation into UU-1093 VL CDR2 to reduce potential Reduces binding of important T cell epitopes, removes oxidation sites, and prevents I to V mutations. Mutations were introduced into UU-1093 VH framework region 3 to enhance potential T cell epitopes. Reduced binding.

[0259] UU-1093-mod6 UU-1093-mod anti-hNAMPT antibody UU-1093-mod6 is SEQ ID NO: 4 a heavy chain variable region having the amino acid sequence shown in SEQ ID NO: 48; The heavy chain variable region of UU-1093-mod6 has the sequence CDR1 domain having the amino acid sequence shown in column number 19, shown in SEQ ID NO: 39 a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 41; The heavy chain of UU-1093-mod6 contains a CDR3 domain that is Mutations were introduced into UU-1093 VH CDR2 to eliminate potential cleavage or fragmentation sites. and an S to T mutation was introduced into UU-1093 VH CDR3 to generate the latent The light chain variable region of UU-1093-mod6 was sequenced as SEQ ID NO: 144444. a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 49; a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 22; The light chain of UU-1093-mod6 contains the CDR3 domain. Natural mutations were introduced into UU-1093 VL CDR1 to reduce or eliminate deamidation. , an I to V mutation was introduced into UU-1093 VL framework region 2 to generate potential Reduced binding of a potential T cell epitope and M-to-V mutation in UU-1093 VL C DR2 to reduce binding of potential T cell epitopes and remove oxidation sites, A I to V mutation was introduced into UU-1093 VH framework region 3 to generate a latent reduced binding of specific T cell epitopes.

[0260] UU-1093-mod7 UU-1093-mod anti-hNAMPT antibody UU-1093-mod7 is SEQ ID NO: 4 a heavy chain variable region having the amino acid sequence shown in SEQ ID NO:2 and a heavy chain variable region having the amino acid sequence shown in SEQ ID NO:48 The heavy chain variable region of UU-1093-mod7 has the sequence CDR1 domain having the amino acid sequence shown in column number 43, shown in SEQ ID NO: 39 a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 44; The heavy chain of UU-1093-mod7 contains a CDR3 domain that converts M to I. The following mutation was introduced into UU-1093 VH CDR1 to remove the oxidation site, changing M to V The following mutations were introduced into the UU-1093 VH framework region 2 to identify the oxidation site and potential The D to E mutation was introduced into UU-1093 VH CDR2 to remove the important T cell epitope. to remove potential cleavage or fragmentation sites, and an M to V mutation was introduced into UU-109 3 VH framework region 3 to remove the oxidation site and a K to R mutation Mutations introduced into UU-1093 VH CDR3 to reduce binding of potential T cell epitopes The light chain variable region of UU-1093-mod7 has the amino acid sequence shown in SEQ ID NO:49. a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 50; a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 51; It comprises a main domain and a CDR3 domain having the amino acid sequence shown in SEQ ID NO: 22. For the light chain of U-1093-mod7, the G to A mutation was replaced with UU-1093 V L CDR1 to reduce or eliminate deamidation, and an I to V mutation to U U-1093 VL framework region 2 to enhance binding of potential T cell epitopes and introducing an M to V mutation into UU-1093 VL CDR2 to reduce potential T Reduced binding of cellular epitopes and eliminated oxidation sites, and I to V mutations UU-1093 introduced into VH framework region 3 to bind potential T cell epitopes was reduced.

[0261] UU-1093-mod8 UU-1093-mod anti-hNAMPT antibody UU-1093-mod8 is SEQ ID NO: 4 a heavy chain variable region having the amino acid sequence shown in SEQ ID NO:5 and a heavy chain variable region having the amino acid sequence shown in SEQ ID NO:48 The heavy chain variable region of UU-1093-mod8 has the sequence CDR1 domain having the amino acid sequence shown in column number 46, shown in SEQ ID NO: 39 a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 41; The heavy chain of UU-1093-mod8 contains a CDR3 domain that is The mutations and M to I were introduced into UU-1093 VH CDR1 and oxidized. The site was removed and an M to V mutation was introduced into UU-1093 VH framework region 2. The D to E mutation was introduced to remove the oxidation site and potential T cell epitopes. 1093 into VH CDR2 to remove potential cleavage or fragmentation sites, and into UU-1093 VH framework region 3 to remove the oxidation site, Then, an S to T mutation was introduced into the UU-1093 VH CDR3 to induce latent T cell proliferation. The light chain variable region of UU-1093-mod8 was sequenced as SEQ ID NO: 49. CDR1 domain having the amino acid sequence shown, amino acid sequence shown in SEQ ID NO: 50 and a CDR3 domain having the amino acid sequence shown in SEQ ID NO: 22. The light chain of UU-1093-mod8 contains a G to A mutation. UU-1093 VL CDR1 to reduce or eliminate deamidation, Mutations to VL were introduced into UU-1093 VL framework region 2 to enhance latent T cell proliferation. Reduced epitope binding and M-to-V mutation in UU-1093 VL CDR2 to reduce binding of potential T cell epitopes and remove oxidation sites, and A VH to VH mutation was introduced into UU-1093 VH framework region 3 to generate a potential T cell Reduced binding of vesicular epitopes.

[0262] UU-1093-mod9 UU-1093-mod anti-hNAMPT antibody UU-1093-mod9 is SEQ ID NO: 3 a heavy chain variable region having the amino acid sequence shown in SEQ ID NO: 8 and a heavy chain variable region having the amino acid sequence shown in SEQ ID NO: 51; The heavy chain variable region of UU-1093-mod9 has the sequence CDR1 domain having the amino acid sequence shown in column number 19, shown in SEQ ID NO: 39 a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 21; The heavy chain of UU-1093-mod9 contains a CDR3 domain that is Mutations were introduced into UU-1093 VH CDR2 to eliminate potential cleavage or fragmentation sites. The light chain variable region of UU-1093-mod9 contains the amino acid sequence shown in SEQ ID NO:11. CDR1 domain having the amino acid sequence shown in SEQ ID NO: 52 2 domain, and a CDR3 domain having the amino acid sequence set forth in SEQ ID NO: 53. For the light chain of UU-1093-mod9, the N to Q mutation was Introducing an I to V mutation into VL CDR1 to reduce or eliminate deamidation was introduced into UU-1093 VL framework region 2 to remove potential T cell epitopes. and an M to V mutation was introduced into UU-1093 VL CDR2 to generate a potential T cell The mutation A to G was introduced to remove the cytoplasmic epitope and the oxidation site. introduced into VL CDR2 to remove potential T cell epitopes and Natural mutations were introduced into UU-1093 VL CDR3 to remove the oxidation site.

[0263] UU-1093-mod10 UU-1093-mod anti-hNAMPT antibody UU-1093-mod10 is identified as SEQ ID NO: 40 and a heavy chain variable region having the amino acid sequence shown in SEQ ID NO: 51. and a light chain variable region having the sequence: a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 19; a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 39; and a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 41. The heavy chain of UU-1093-mod10 contains a CDR3 domain having the sequence D. A mutation from α to E was introduced into UU-1093 VH CDR2 to prevent potential truncation or fragmentation. The site was removed and an S to T mutation was introduced into UU-1093 VH CDR3. The light chain variable region of UU-1093-mod10 was a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 11; a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 52; and a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 53. The light chain of UU-1093-mod10 contains a CDR3 domain with the following structure: Introducing a Q to Q mutation into UU-1093 VL CDR1 reduces deamidation or removed and an I to V mutation introduced into UU-1093 VL framework region 2 to remove potential T cell epitopes and the M to V mutation in UU-1093 VL CDR2 to remove potential T cell epitopes and to remove oxidation sites, A mutation from α to G was introduced into UU-1093 VL CDR2 to identify potential T cell epitopes. The fragment was removed and a W to F mutation was introduced into UU-1093 VL CDR3. The oxidation sites were removed.

[0264] UU-1093-mod11 UU-1093-mod anti-hNAMPT antibody UU-1093-mod11 is identified as SEQ ID NO: a heavy chain variable region having the amino acid sequence set forth in SEQ ID NO: 42 and a heavy chain variable region having the amino acid sequence set forth in SEQ ID NO: 51; and a light chain variable region having the sequence: a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 43; a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 39; and a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 44. The heavy chain of UU-1093-mod11 contains a CDR3 domain having the sequence M. A mutation from M to I was introduced into UU-1093 VH CDR1 to remove the oxidation site, and Mutations from V to V were introduced into the UU-1093 VH framework region 2 to locate the oxidation site and The D to E mutation was introduced into UU-1093 VH C to remove potential T cell epitopes. DR2 to remove a potential cleavage or fragmentation site, and an M to V mutation was introduced into UU- 1093 into VH framework region 3 to remove the oxidation site and Mutations were introduced into UU-1093 VH CDR3 to block potential T cell epitope binding. The light chain variable region of UU-1093-mod11 is the amino acid sequence shown in SEQ ID NO: 11. a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 52; a CDR2 domain and a CDR3 domain having the amino acid sequence shown in SEQ ID NO: 53 The light chain of UU-1093-mod11 contains an N to Q mutation. 093 VL CDR1 to reduce or eliminate deamidation and to A natural mutation was introduced into UU-1093 VL framework region 2 to identify potential T cell epitopes. The fragment was removed and an M to V mutation was introduced into UU-1093 VL CDR2 to generate potential The mutation A to G was introduced to remove the T cell epitope and the oxidation site. 093 VL CDR2 to remove potential T cell epitopes, and W to F A mutation to was introduced into UU-1093 VL CDR3 to remove the oxidation site.

[0265] UU-1093-mod12 UU-1093-mod anti-hNAMPT antibody UU-1093-mod12 is identified as SEQ ID NO: a heavy chain variable region having the amino acid sequence set forth in SEQ ID NO: 45 and a heavy chain variable region having the amino acid sequence set forth in SEQ ID NO: 51; and a light chain variable region having the sequence: a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 46; a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 39; and a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 41. The heavy chain of UU-1093-mod12 contains a CDR3 domain having the sequence W. A nucleotide to F mutation and an M to I mutation were introduced into UU-1093 VH CDR1. to remove the oxidation site and M to V mutations in the UU-1093 VH framework region. 2 to remove the oxidation site and potential T cell epitopes, and a D to E mutation introduced into UU-1093 VH CDR2 to remove potential cleavage or fragmentation sites, A VH mutation was introduced into UU-1093 VH framework region 3 to remove the oxidation site. and an S to T mutation was introduced into UU-1093 VH CDR3 to enhance potential The light chain variable region of UU-1093-mod12 is SEQ ID NO: a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 11; a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 53; The light chain of UU-1093-mod12 contains the CDR3 domain. Mutations introduced into UU-1093 VL CDR1 to reduce or eliminate deamidation and introducing an I to V mutation into UU-1093 VL framework region 2 to generate latent The M to V mutation in the UU-1093 VL CDR removed the target T cell epitope. 2 to remove potential T cell epitopes and to remove oxidation sites, and Mutations introduced into UU-1093 VL CDR2 to remove potential T cell epitopes and a W to F mutation was introduced into UU-1093 VL CDR3 to remove the oxidation site. was removed.

[0266] UU-1093-mod13 The UU-1093-mod anti-hNAMPT antibody UU-1093-mod13 is represented by SEQ ID NO: a heavy chain variable region having the amino acid sequence shown in SEQ ID NO: 38 and a heavy chain variable region having the amino acid sequence shown in SEQ ID NO: 18 and a light chain variable region having the sequence: a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 19; a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 39; and a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 21. The heavy chain of UU-1093-mod13 contains a CDR3 domain having the sequence D. A mutation from α to E was introduced into UU-1093 VH CDR2 to prevent potential truncation or fragmentation. The light chain variable region of UU-1093-mod13 is shown in SEQ ID NO:6. a CDR1 domain having the amino acid sequence shown in SEQ ID NO:7; a CDR2 domain and a CDR3 domain having the amino acid sequence shown in SEQ ID NO: 22 The light chain of UU-1093-mod13 is identical to UU-1093 VL.

[0267] UU-1093-mod14 The UU-1093-mod anti-hNAMPT antibody UU-1093-mod14 is identified as SEQ ID NO: 40 and a heavy chain variable region having the amino acid sequence shown in SEQ ID NO: 18. and a light chain variable region having the sequence: a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 19; a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 39; and a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 41. The heavy chain of UU-1093-mod14 contains a CDR3 domain having the sequence D. A mutation from α to E was introduced into UU-1093 VH CDR2 to prevent potential truncation or fragmentation. The site was removed and an S to T mutation was introduced into UU-1093 VH CDR3. The light chain variable region of UU-1093-mod14 was , a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 6, a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 7 a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 22; The light chain of UU-1093-mod14 contains the CDR3 domain that binds to UU-1093. Identical to VL.

[0268] UU-1093-mod15 The UU-1093-mod anti-hNAMPT antibody UU-1093-mod15 is identified as SEQ ID NO: 42 and a heavy chain variable region having the amino acid sequence shown in SEQ ID NO: 18. and a light chain variable region having the sequence: a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 43; a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 39; and a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 44. The heavy chain of UU-1093-mod15 contains a CDR3 domain having the sequence M. A mutation from M to I was introduced into UU-1093 VH CDR1 to remove the oxidation site, and Mutations from V to V were introduced into the UU-1093 VH framework region 2 to locate the oxidation site and The D to E mutation was introduced into UU-1093 VH C to remove potential T cell epitopes. DR2 to remove a potential cleavage or fragmentation site, and an M to V mutation was introduced into UU- 1093 into VH framework region 3 to remove the oxidation site and Mutations were introduced into UU-1093 VH CDR3 to block potential T cell epitope binding. The light chain variable region of UU-1093-mod15 is the amino acid sequence shown in SEQ ID NO:6. a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 7; 2 domain, and a CDR3 domain having the amino acid sequence set forth in SEQ ID NO: 22. The light chain of UU-1093-mod15 is identical to UU-1093 VL.

[0269] UU-1093-mod16 The UU-1093-mod anti-hNAMPT antibody UU-1093-mod16 is identified as SEQ ID NO: 45 and a heavy chain variable region having the amino acid sequence shown in SEQ ID NO: 18. and a light chain variable region having the sequence: a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 46; a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 39; and a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 41. The heavy chain of UU-1093-mod16 contains a CDR3 domain having the sequence W. A nucleotide to F mutation and an M to I mutation were introduced into UU-1093 VH CDR1. to remove the oxidation site and M to V mutations in the UU-1093 VH framework region. 2 to remove the oxidation site and potential T cell epitopes, and a D to E mutation introduced into UU-1093 VH CDR2 to remove potential cleavage or fragmentation sites, A VH mutation was introduced into UU-1093 VH framework region 3 to remove the oxidation site. and an S to T mutation was introduced into UU-1093 VH CDR3 to enhance potential The light chain variable region of UU-1093-mod16 is SEQ ID NO: No. 6, a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 7 a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 22; The light chain of UU-1093-mod16 contains the same VL as UU-1093. It is one.

[0270] UU-1093-mod17 The UU-1093-mod anti-hNAMPT antibody UU-1093-mod17 is identified as SEQ ID NO: 26 and a heavy chain variable region having the amino acid sequence shown in SEQ ID NO: 47. and a light chain variable region having the sequence: a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 19; a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 20; and a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 21. The heavy chain of UU-1093-mod17 contains a CDR3 domain having the sequence 3 VH. The light chain variable region of UU-1093-mod17 is set forth in SEQ ID NO: 11. a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 7; and a CDR3 domain having the amino acid sequence shown in SEQ ID NO: 22. The light chain of UU-1093-mod17 contains an N to Q mutation. Deamidation was reduced or eliminated by introducing it into UU-1093 VL CDR1.

[0271] UU-1093-mod18 The UU-1093-mod anti-hNAMPT antibody UU-1093-mod18 is identified as SEQ ID NO: 26 and a heavy chain variable region having the amino acid sequence shown in SEQ ID NO: 48. and a light chain variable region having the sequence: a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 19; a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 20; and a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 21. The heavy chain of UU-1093-mod18 contains a CDR3 domain having the sequence 3 VH. The light chain variable region of UU-1093-mod18 is set forth in SEQ ID NO: 49. CDR1 domain having the amino acid sequence shown, amino acid sequence shown in SEQ ID NO: 50 and a CDR3 domain having the amino acid sequence shown in SEQ ID NO: 22. The light chain of UU-1093-mod18 contains a G to A mutation. was introduced into UU-1093 VL CDR1 to reduce or eliminate deamidation, A VL to VL mutation was introduced into UU-1093 VL framework region 2 to generate a potential T cell The M to V mutation in UU-1093 VL CDR2 reduces binding of the cytoplasmic epitope. to reduce binding of potential T cell epitopes and remove oxidation sites, and A potential T to V mutation was introduced into UU-1093 VH framework region 3 to Reduced binding of cellular epitopes.

[0272] UU-1093-mod19 The UU-1093-mod anti-hNAMPT antibody UU-1093-mod19 is identified as SEQ ID NO: 26 and a heavy chain variable region having the amino acid sequence shown in SEQ ID NO: 51. and a light chain variable region having the sequence: a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 19; a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 20; and a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 21. The heavy chain of UU-1093-mod17 contains a CDR3 domain having the sequence 3 VH. The light chain variable region of UU-1093-mod19 is set forth in SEQ ID NO: 11. CDR1 domain having the amino acid sequence shown, amino acid sequence shown in SEQ ID NO: 52 and a CDR3 domain having the amino acid sequence shown in SEQ ID NO: 53. The light chain of UU-1093-mod19 contains an N to Q mutation. was introduced into UU-1093 VL CDR1 to reduce or eliminate deamidation, A VL to VL mutation was introduced into UU-1093 VL framework region 2 to generate a potential T cell Removed the cytoplasmic epitope and introduced an M to V mutation into UU-1093 VL CDR2 to remove potential T cell epitopes and to remove the oxidation site, and A to G mutation into UU-1093 VL CDR2 to remove potential T cell epitopes, and A W to F mutation was introduced into the UU-1093 VL CDR3 to remove the oxidation site. Ta.

[0273] [Table 32] [Table 33] [Table 34] [Table 35] [Table 36] [Table 37] [Table 38] [Table 39] [Table 40] [Table 41] [Table 42] [Table 43] In Table 14, the mutated residues are shown in bold.

[0274] [Table 44] [Table 45] [Table 46] [Table 47]

[0275] [Table 48] [Table 49] [Table 50] [Table 51]

[0276] Example 12. Tissue expression of NAMPT in human invasive PCa To evaluate the role of NAMPT in the invasiveness and progression of prostate cancer (PCa), The expression of AMPT was studied in PCa tissues.

[0277] Normal prostate tissue, prostate adenocarcinoma confined to the prostate without capsular invasion, and periprostatic Immunohistochemical (IHC) staining of prostate adenocarcinoma with capsular invasion into peri-adipose tissue Representative photomicrographs are provided in Figures 9A to 9C. Furthermore, a group of PCa patients with benign prostate tissue, organ-confined disease (T2 disease, n=12) was included. IH in tissues from PCa patients with tissue and capsular invasive disease (T3 disease, n=14) NAMPT expression was assessed by C staining, and the cumulative analysis is provided in Figure 9D.

[0278] As shown in Figure 9, IHC analysis of normal and PCa tissues showed that normal prostate tissue In the 1990s, there was virtually no NAMPT expression (Fig. 9A), which was localized to the prostate but without capsular invasion. Prostate adenocarcinoma showed minimal expression of NAMPT (Figure 9B). Prostate adenocarcinoma with capsular invasion into periprostatic adipose tissue showed significantly stronger NAMPT staining. (Figure 9C). In addition, 26 PCa patients with benign prostate tissue and T2 and T3 disease Comparative analysis of tissues from the study showed increased expression of NAMPT with increasing PCa invasiveness. Therefore, increased expression of NAMPT was associated with the development of human invasive P. It was observed in Ca.

[0279] Example 13. Effect of radiation exposure on NAMPT expression Radiation therapy is the mainstay of PCa therapy. It functions as a damage-associated molecular pattern protein (DAMP) in the immune system, By assessing the expression of NAMPT in mouse and human tissues exposed to 1000 r.i.m. , the effect of radiation on NAMPT expression was evaluated.

[0280] Effect of radiation-induced tissue injury and damage on NAMPT expression in mouse tissues To evaluate this, C57 / B6 mice were exposed to a single dose of thoracic radiation (20 Gy) for 1 week. The effects of radiation on inflammation, blood leakage, and inflammatory lung injury were examined in mice after radiation exposure. Lung tissue was assessed by H&E staining, and representative photomicrographs are provided in Figure 10A. The expression of NAMPT was evaluated in lung tissue before and after irradiation by IHC staining. Micrographs are provided in Figures 10B and 10C. To evaluate the effect of radiation on normal human epithelial tissue (tonsils), we irradiated (8 Gy). After 24 hours of exposure, NAMPT expression was assessed by IHC staining. Provided in Figure 10D.

[0281] As shown in Figure 10, after one week of radiation exposure, inflammation, blood leakage, and inflammation were observed in the lung tissue of the mice. Increased pathogenic damage was observed (Fig. 10A). Radiation-induced lung injury was particularly pronounced in the alveolar macrophages. There was significant NAMPT expression (Fig. 10B) in the oocytes and epithelial cells (Fig. 10C). Consistent with radiation as a stimulus for NAMPT tissue expression in mice, 24 h of radiation exposure A significant increase in NAMPT expression was observed in normal human epithelial tissue (tonsil) after 2 h (Figure 1). 10D). Thus, radiation-induced tissue damage significantly induced NAMPT expression.

[0282] Example 14. Role of NAMPT in PCa cell migration Using an in vitro assay of PCa cell migration through human smooth muscle cells, PCa cells We evaluated the role of NAMPT in migration of .

[0283] To evaluate the role of NAMPT in PCa cell migration, NAMPT (100 ng 1 mL) was added to cultures of human DU-145 PCa cells. 24 hours after NAMPT exposure PCa cell migration was assessed. A summary of the observations is provided in Figure 11A, and representative micrographs are shown. The truth is provided in Figure 11B.

[0284] As observed in Figure 11, NAMPT potently stimulated the growth of human DU-145 PCa cells. It functioned as a chemoattractant and induced PCa cell migration for 24 hours.

[0285] Example 15. Characterization of humanized anti-hNAMPT antibodies 1076 Humanized anti-hNAMPT antibodies (N-1076, K-1076 and P-1076) and 1093 humanized anti-hNAMPT antibodies (SS-1093, XX-1093, and UU-1 093) to treat lung injury in mice induced by intratracheal delivery of LPS. "One-hit" models of injury and the use of LPS and mechanical VILI in mice Using two mouse lung injury models, To assess the ability of the antibodies to reduce acute inflammation and injury, humanized These mice were administered either the anti-hNAMPT antibody or the anti-hNAMPT antibody. The results of the study are presented in Figure 12. It is served.

[0286] As shown in Figure 12A, analysis of the integrated lung injury score showed that all anti-hNA MPT antibody is effective in reducing lung injury in an LPS-induced "one-hit" model However, the most substantial effect was observed with the anti-hNAMPT antibody P-1076. As shown in Figure 12B, analysis of the integrated lung injury score showed that All anti-hNAMPT antibodies tested suppressed lung injury in the LPS / VILI-induced "two-hit" model. However, the most substantial effect was seen in anti-hN As shown in Figure 12C, the anti-hNAMPT antibody P-1076 Antibody P-1076 inhibits the synthesis of inflammatory cytokines in an LPS / VILI-induced "two-hit" model of acute inflammatory injury. It is effective in reducing the histological damage index.

[0287] Example 16. Effect of humanized anti-hNAMPT antibody P-1076 on PCa cell invasion To evaluate the effect of humanized anti-hNAMPT antibody P-1076 on PCa cell invasion We evaluated peritoneal invasion of human PCa cells in mice with severe combined immunodeficiency (SCID).

[0288] To evaluate the role of humanized anti-hNAMPT antibody P-1076 on PCa cell invasion Next, metastatic human PCa cells PC3 were injected intraperitoneally (IP) into SCID mice. The mice were injected twice weekly with 2 μg of the humanized anti-hNAMPT antibody P-1076 or vehicle alone. Six weeks after injection, peritoneal infiltration of PC3 cells was evaluated. Representative micrographs A summary of the results is provided in FIG.

[0289] As shown in Figure 13, injection of human PCa cell lines resulted in substantial peritoneal muscle invasion. In contrast, the humanized anti-hNAMPT antibody P-1076 was administered to C3-challenged SCID mice showed a marked reduction in PC3 infiltration into the smooth muscle peritoneum (Figure 13B).

[0290] Thus, as summarized in Figure 13C, the observations from this study suggest that the invasion of PCa cells The role of NAMPT in invasiveness and the use of humanized anti-hNAMPT antibodies to transiently induce this invasive behavior This strongly suggests the important potential of entity P-1076.

[0291] Example 17. In vivo treatment of RILI using anti-NAMPT antibodies The ability of anti-NAMPT administration to affect RILI was investigated in vivo in C57 / B6 mice. Mice received 20 Gy of thoracic radiation and were administered polyclonal NAM. Mice were intraperitoneally injected with PT neutralizing antibodies (pAb) and received 20 Gy of thoracic radiation. Monoclonal anti-NAMPT antibody (mAb) (P-1076-mod1) was injected intraperitoneally. irradiated mice injected with vehicle only ("Ctrl"), and non-irradiated mice injected with vehicle only ("Ctrl"). The mice were divided into four groups: irradiated mice injected with only BAL protein ("Ctrl"), and irradiated mice injected with only BAL protein ("Ctrl"). The amount of BAL-expressing cells was measured and the number of BAL-expressing cells was obtained. Lung tissue was also subjected to H&E staining to determine the extent of lung inflammation. Furthermore, the acute lung injury (ALI) severity score was evaluated based on the BAL index and H&E staining. The results of the corresponding analyses are provided in Figures 14A-E.

[0292] As shown in Figure 14A, lung tissue from a non-irradiated control mouse (insert in the left panel of Figure 14A) H&E staining of lung tissue from irradiated control mice (injected with vehicle only) compared with that from irradiated control mice (Figure 1). The color indicated diffuse alveolar damage 4 weeks after radiation exposure (left panel of Figure 14A). In contrast, anti-NAMPT pAb (middle panel of Figure 14A) or anti-NAMPT mAb (Figure Lung tissue from mice injected with IFN-γ (right panel of 14A) showed reduced H&E staining and showed that mice treated with anti-NAMPT Ab after radiation exposure had less alveolar damage. Figure 14B shows the results of non-irradiated control mice, irradiated control mice, and mice treated with anti-NAMPT pAb or m Figure 14 summarizes H&E staining in lung tissues of irradiated mice injected with Ab. As shown in B, the H&E stained area was significantly higher in the irradiated control mice compared to the non-irradiated control mice. However, the anti-NAMPT A significant decrease in H&E stained areas was observed in lung tissue from mice injected with pAb or mAb. (p<0.05), suggesting a role for NAMPT in the pathogenesis of RILI. 4C shows non-irradiated control mice, irradiated control mice, and mice injected with anti-NAMPT pAb or mAb. Figure 1 summarizes BAL protein levels in lung tissue of irradiated mice. As shown in 14C, compared with non-irradiated control mice, radiation-exposed mice showed significantly increased BA However, anti-NAMPT pAb or mAb showed an increase in L protein levels. Injected irradiated mice showed significant increases in BAL protein levels compared to irradiated control mice. (p<0.05), and was significantly reduced in irradiated mice treated with anti-NAMPT mAb. Similarly, the number of BAL cells increased in mice exposed to irradiation. However, irradiated mice injected with anti-NAMPT pAb or mAb showed similar growth rates to irradiated control mice. In comparison, the number of BAL cells was significantly reduced (p<0.05), and anti-NAMPT m A more significant decrease was observed in irradiated mice treated with Ab. Furthermore, as shown in Figure 14E, Furthermore, compared with control mice, radiation-exposed mice showed an increase in ALI severity scores. As shown, irradiated mice injected with anti-NAMPT pAb or mAb were significantly higher than irradiated control mice. The ALI severity score was significantly reduced compared with the control group, and the anti-NAMPT mAb treatment A more significant decrease was observed in irradiated mice treated with anti-N. This indicates a reduction in RILI after treatment with AMPT Ab, and is a potential therapeutic target for RILI. We emphasize NAMPT as a target.

[0293] Example 18. Radiolabeled anti-NAM antibody detects increased NAMPT expression in inflamed lung tissue Identification by PT antibody Radiolabeled anti-NAMPT antibodies were used to investigate the NAMPT signaling pathway and It was developed to detect NAMPT expression non-invasively in vivo. Mice with RILI were treated with anti-NAMPT mAb (P-1076-mod1). Optimal time to image the model and grow anti-NAMPT mAb as therapeutic intervention and after total body irradiation (TBI) or partial body irradiation (PBI) in a nuclear accident, Other specific radiolabels can be used to determine the role of inflammatory apoptosis and cellular apoptosis. Organs can be investigated. Detection of NAMPT expression using radiolabeled anti-NAMPT antibody To test the 99m Tc-labeled anti-NAMPT mAb probe was administered to control mice and 8 Mice were injected with 1000 Gy of PBI and rapid autoradiographic imaging was performed. The results of the analysis are shown in Figures 15A-D.

[0294] As shown in Figures 15A-B, the lungs of irradiated mice showed higher Radioactive uptake was observed, indicating higher NAMPT expression induced by RILI. Furthermore, the uptake of radiolabeled anti-NAMPT antibody was measured in irradiated mice or non-irradiated control mice. As shown in Figure 15C, right lung activity from a non-irradiated control mouse was used as a reference for lung activity in the control mouse. The tissue background was significantly higher in both the right and left lungs from irradiated mice compared with the left and right lungs. A significant increase in lung activity was observed (p<0.05). The level of radioactivity in illuminated mice was determined to assess the uptake of radiolabeled anti-NAMPT mAb. As shown in Figure 15D, the radiation dose was significantly higher in irradiated mice compared to non-irradiated control mice. A significant increase in the activity of radiolabeled anti-NAM in irradiated mice was observed (p<0.05). Increased uptake of PT mAb was confirmed.

[0295] Therefore, radiolabeled anti-NAMPT antibodies could detect NAMPT in inflamed lung tissue. This was due to the fact that radiolabeled anti-NAMPT antibodies were effective in detecting increased NAMPT expression. We emphasized the possibility of using AMPT as a detection tool and as a biomarker for RILI. This may be crucial for the use of NAMPT.

[0296] Example 19. Therapeutic Targeting of RILI Using an In Vivo Model of Radiation-Induced Pulmonary Fibrosis Validation of NAMPT To further validate NAMPT as a therapeutic target for RILI, we used WT C57 / B6 mice. The mice were exposed to 20 Gy WT1. Irradiated mice were treated with 10 μg of anti-NAMPT mAb ( P-1076-mod1) or vehicle control was injected intraperitoneally. BAL cell count, collagen The accumulation of pulmonary smooth muscle actin (SMA) in the lung is a reflection of myofibroblast migration and fibrosis. Radiation-induced pulmonary fibrosis (RILF) was assessed 18 weeks after radiation exposure by assessing the expression of The mice were evaluated for serotonin-dependent serotonin (SDS) and serotonin-dependent serotonin (SDS). The results are shown in Figures 16A-C.

[0297] As shown in Figure 16A-C, Ab-treated mice showed significantly higher IFN-γ levels than vehicle-treated control mice. In mice treated with NAMPT, anti-NAMPT mAb significantly reduced IR-induced RILI, and this reduction The results showed a decrease in BAL cell count (Fig. 16A), a decrease in SMA expression in lung tissue (Western blot analysis). analysis and shown in Figure 16B), and a reduction in collagen deposition (trichloroquine in lung tissue). This is reflected in the chromatin staining (detected by chromatin staining and shown in Figure 16C).

[0298] Therefore, the results emphasized the role of anti-NAMPT Ab in alleviating RILF and R Further validate NAMPT as a therapeutic target for ILI.

[0299] Example 20. Evaluation of the efficacy of anti-NAMPT mAb in preclinical models of lung injury Anti-NAMPT in a rat model of trauma (blast injury) / ventilator-induced lung injury (VILI) The efficacy of the mAb was verified by challenging Sprague-Dawley rats with trauma (blast injury). After the blast injury, rats were administered anti-NAMPT mAb (P- 100 μg of 1076-mod1 was injected intravenously (IV). Exposed rats injected with vehicle served as controls. After 10 min, lungs were removed from the rats and assessed for damage. H&E staining of lung tissue revealed edema and inflammatory cell infiltration, which are used to interpret lung injury. The results of this trauma (blast injury) / VILI lung injury model are shown in Figures 17A-C. Provided.

[0300] As shown in Figure 17A, unchallenged rats (insert in the right-most box in Figure 17A) Compared to the lung tissue from vehicle-injected control trauma / VILI rats, the lung tissue from the control trauma / VILI rats showed significantly less inflammation than that from the control trauma / VILI rats. Trauma / VILI-induced lung injury is demonstrated, showing cellular infiltration and edema. As shown in 17B, lungs from trauma / VILI rats treated with anti-NAMPT mAb The tissue showed a significant reduction in inflammatory cell infiltration and edema, suggesting that anti-NAMPT mAb Attenuation of trauma / VILI-induced lung injury by IVF therapy. The effect of anti-NAMPT mAb on the proliferation and proliferation of NAMPT in rat lung tissue was assessed by H&E staining index. The injury scores are summarized in Figure 17C. As shown in Figure 17C, the vehicle control The lung injury scores were significantly higher in rats treated with anti-NAMPT mAb compared with rats treated with IFN-α. (p<0.05). Therefore, the results outlined in Figures 17A-C suggest that the Figure 1 shows the efficacy of NAMPT-neutralizing mAb in attenuating VILI-induced lung injury.

[0301] Next, the efficacy of anti-NAMPT mAb was tested in a mouse model of LPS / VILI. Mice were challenged with LPS for 18 hours and then mechanically ventilated for 4 hours. MPT mAb (P-1076-mod1, 10 μg, IV), anti-NAMPT polyclonal antibody Injections of either parental antibody (pAb) or vehicle control (PBS) were administered. Mice without the steroid agent served as controls. Lung tissue from the mice was then analyzed by H&E staining. Edema and inflammatory cell infiltration in the lungs were assessed as a readout of lung injury. The results of the ILI lung injury model are provided in Figures 18A-C.

[0302] As shown in Figure 18A, compared to unchallenged mice (inset in Figure 18A), Lung tissue from control mice injected with LPS / V showed inflammatory cell infiltration and edema, whereas LPS / V In contrast, as shown in Figure 18B, anti-NAMPT cells inhibited ILI-induced lung injury. Lung tissue from mAb-treated mice showed a significant reduction in inflammatory cell infiltration and edema. Therefore, this study demonstrated that anti-NAMPT mAb attenuated LPS / VILI-induced lung injury. The effect of anti-NAMPT mAb on trauma / VILI-induced lung injury was investigated by H&E staining. Figure 18C shows the acute lung injury (ALI) severity score of mice as assessed by the number of serotonin-positive and serotonin-negative lungs. As shown in Figure 18C, compared to vehicle-injected mice, anti-NAM ALI was significantly reduced in mice treated with PT pAb or mAb, and anti-NAMPT m The strongest reduction in ALI severity scores was observed in Ab-treated mice (p<0. 001). Therefore, the results outlined in Figures 18A-C suggest that trauma / VILI-induced lung 1 shows the efficacy of NAMPT neutralizing mAb in reducing damage.

[0303] Thus, these results support the role of IL-1 in reducing lung injury in a preclinical in vivo lung injury model. This demonstrates the efficacy of anti-NAMPT mAb.

[0304] Example 21. Radiolabeled anti-NAM antibody detects increased NAMPT expression in inflamed lung tissue Identification by PT antibody Humanized anti-NAMPT mAb (K-1076) was radiolabeled to investigate its activity in various tissues. The NAMPT signaling pathway and NAMPT expression can be detected noninvasively in vivo. We have developed an imaging probe that can detect acute inflammatory diseases (e.g., COVID-19, NAMPT as a diagnostic and / or prognostic biomarker in ARDS and lung injury Considering this possibility, radiolabeled anti-NAMPT mAb may be useful for reducing the risk of developing such diseases. It may be used as a diagnostic tool in at-risk subjects or by anti-NAMPT mAb. It can be used to select subjects who are likely to respond to treatment of such inflammatory conditions. In this example, radiolabeled anti-NAMPT mAb was used to assess the effect of LPS challenge and ionization. The detection of NAMPT expression in inflamed tissues such as radiation-exposed lungs is described.

[0305] First, to test the detection of NAMPT expression using radiolabeled anti-NAMPT antibody, 9 9m Tc-labeled anti-NAMPT mAb probe or radiolabeled IgG control Ab was administered at 20 Gy total dose. Immediate autoradiographic images of mice exposed to thoracic lung irradiation (WTLI) We conducted a survey.

[0306] As shown in Figure 19A, irradiated mice injected with radiolabeled IgG control (left panel of Figure 19A) Compared with the side panel, radiolabeled anti-NAMPT mAb (PRONAMPTOR) was injected. Significantly higher radioactive uptake was observed in the irradiated mice (right panel of Figure 19A). Therefore, the results shown in Figure 19A indicate that radiation-induced NAMPT expression is The ability of radiolabeled anti-NAMPT imaging probes is demonstrated.

[0307] Further evaluation of the detection of NAMPT expression by radiolabeled anti-NAMPT imaging probes In order to 99mTc-labeled anti-NAMPT mAb was administered to vehicle-challenged control mice or LPS-challenged mice were injected 3 or 18 hours after LPS challenge, and rapid onset was observed. Radiographic imaging was performed, and the results of the analysis are shown in Figures 19B-D.

[0308] As shown in Figure 19B, LPS-challenged mice were significantly more susceptible to steroids than control mice (left panel of Figure 19B). Range mice were treated with radiolabeled anti-NAMPT imaging probes 3 hours after LPS challenge. LPS-challenged mice showed significantly higher uptake of the IgG lobes (right panel of Figure 19B). Autoradiographic imaging of lungs from bronchial or control mice further confirmed this observation. pulmonary leukemia in LPS-challenged mice compared to control mice (left panel of Figure 19C). showed a significant increase in the activity of the radiolabeled anti-NAMPT imaging probe 3 hours after LPS challenge. Furthermore, as shown in Figure 19D, the control showed significantly higher uptake than the control (right panel of Figure 19C). Compared with mice, LPS-challenged mice showed significantly higher IL-1 levels at 3 and 18 hours after LPS challenge. Afterward, significantly higher radioactivity was observed (p<0.05), and radiolabeled anti-NAMPT imaging Therefore, the uptake of the lobes is high. The results showed that radiolabeled anti-NAMPT imaging was effective in detecting LPS-induced NAMPT expression. This demonstrates the capabilities of the ing probe.

[0309] Therefore, radiolabeled anti-NAMPT antibody inhibits NAMPT expression in inflamed tissues. This was because radiolabeled anti-NAMPT antibodies were effective in detecting increases in NAMP. highlighting its potential use as a tool for the detection of T in acute inflammatory diseases and / or This may be crucial for the use of NAMPT as a prognostic biomarker. We then investigated the efficacy of this radiolabeled anti-NAMPT antibody by detecting increased NAMPT expression in inflamed tissues. The AMPT imaging probe is a promising tool for the treatment of acute inflammatory diseases with neutralizing anti-NAMPT mAb. It can be used to select subjects who are more likely to respond to a treatment.

[0310] Example 22. Reduction of PAH manifestations in a rat model by anti-NAMPT antibodies To explore the potential of NAMPT as a therapeutic target for PAH, we performed a study on rat monoclonal antibodies against PAH. A crotaline (MCT) model was used. A single dose (60 mg / kg body weight) of MCT was administered. Subcutaneous injection was performed on Larg-Dawley rats (190-200 g). The rats were treated with anti-NAMPT mAb (P-1076-mod1, i.p., 100 μg / rat). The rats were then injected twice a week with either IFN-γ-glucan (IFN-γ rats) or vehicle control (control MCT rats). Right ventricular systolic pressure (RVSP) and pulmonary artery remodeling were assessed in the rats. The results are shown in Figure 20A and 20B. and 20B.

[0311] Right heart catheterization using a Millar pressure transducer catheter RVSP was measured in MCT rats treated with anti-NAMPTAb or control MCT rats. As shown in Figure 20A, the anti-NAMPT mAb-treated rats showed significantly higher leukemia rates compared to the control MCT rats. A significant decrease in RVSP was observed in MCT rats treated with IVF (p<0.05).

[0312] H&E of lungs from MCT rats treated with anti-NAMPT Ab or control MCT rats After staining with , pulmonary artery remote imaging was performed using APERIO IMAGESCOPE software. As shown in Figure 20B, the anti-NAMP A significant decrease in pulmonary artery thickness was observed in MCT rats treated with T mAb.

[0313] The results showed that neutralization of NAMPT by anti-NAMPT mAb significantly improved PAH in a rat model. to reverse vascular remodeling and RV dysfunction in PAH. The effectiveness of NAMPT as a

[0314] Unless otherwise stated, the disclosures of all patents, patent applications and publications cited herein are incorporated by reference. The contents of which are incorporated herein by reference to the extent that the reference describes information relevant to this disclosure. Although the present invention has been disclosed with reference to specific embodiments, some embodiments of the present invention may and modifications may be devised by those skilled in the art without departing from the true spirit and scope of the invention. It is apparent that the appended claims encompass all such embodiments and equivalents. This includes variations such as the above.

[0315] The sequences disclosed herein and / or related to the present invention are set forth in the Sequence Summary Table below. (Table 17).

[0316] [Table 52] [Table 53] [Table 54] [Table 55] [Table 56] Table 57 Table 58 Table 59 Table 60 Table 61 Table 62

Claims

1. Isolation of binding to human nicotinamide phosphoribosyltransferase (NAMPT) an antibody or antigen-binding fragment thereof, (i) a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 3, SEQ ID NO: 4, or 29, and a CDR2 domain having the amino acid sequence shown in SEQ ID NO:

5. a heavy chain variable region comprising a CDR3 domain having the sequence (ii) a CDR1 domain having the amino acid sequence set forth in SEQ ID NO: 6 or 11, the sequence CDR2 domain having the amino acid sequence shown in No. 7, 12, 14, 33, 35 or 37 and a light chain variable domain comprising a CDR3 domain having the amino acid sequence set forth in SEQ ID NO:

8. and a region.

2. The heavy chain variable region (a) a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 3, a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 4 and a CDR2 domain having the amino acid sequence shown in SEQ ID NO:

5. or a CDR3 domain having (b) a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 3, a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 29 and a CDR2 domain having the amino acid sequence shown in SEQ ID NO:

5.

2. The isolated antibody or antigen-binding thereof of claim 1, comprising a CDR3 domain having a sequence Fragment.

3. The light chain variable region (a) a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 6, a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 7, and a CDR2 domain having the amino acid sequence shown in SEQ ID NO:

8. a CDR3 domain having (b) a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 6, a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 12 and a CDR2 domain having the amino acid sequence shown in SEQ ID NO:

8. a CDR3 domain having a sequence (c) a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 6, a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 14 and a CDR2 domain having the amino acid sequence shown in SEQ ID NO:

8. a CDR3 domain having a sequence (d) a CDR1 domain having the amino acid sequence set forth in SEQ ID NO: 6, a CDR2 domain having the amino acid sequence set forth in SEQ ID NO: 33 and a CDR2 domain having the amino acid sequence shown in SEQ ID NO:

8. a CDR3 domain having a sequence (e) a CDR1 domain having the amino acid sequence set forth in SEQ ID NO: 6, a CDR2 domain having the amino acid sequence set forth in SEQ ID NO: 35 and a CDR2 domain having the amino acid sequence shown in SEQ ID NO:

8. a CDR3 domain having a sequence (f) a CDR1 domain having the amino acid sequence set forth in SEQ ID NO: 6, a CDR2 domain having the amino acid sequence set forth in SEQ ID NO: 37 and a CDR2 domain having the amino acid sequence shown in SEQ ID NO:

8. a CDR3 domain having a sequence (g) a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 11, and a CDR2 domain having the amino acid sequence shown in SEQ ID NO:

8. a CDR3 domain having a sequence (h) a CDR1 domain having the amino acid sequence set forth in SEQ ID NO: 11, SEQ ID NO: 12 and a CDR2 domain having the amino acid sequence shown in SEQ ID NO:

8. a CDR3 domain having the sequence (i) a CDR1 domain having the amino acid sequence set forth in SEQ ID NO: 11, SEQ ID NO: 14 and a CDR2 domain having the amino acid sequence shown in SEQ ID NO:

8. a CDR3 domain having the sequence (j) a CDR1 domain having the amino acid sequence set forth in SEQ ID NO: 11, SEQ ID NO: 33 and a CDR2 domain having the amino acid sequence shown in SEQ ID NO:

8. a CDR3 domain having the sequence (k) a CDR1 domain having the amino acid sequence set forth in SEQ ID NO: 11, SEQ ID NO: 35 and a CDR2 domain having the amino acid sequence shown in SEQ ID NO:

8. a CDR3 domain having the sequence (l) a CDR1 domain having the amino acid sequence set forth in SEQ ID NO: 11, SEQ ID NO: 37 and a CDR2 domain having the amino acid sequence shown in SEQ ID NO:

8.

3. The isolated antibody or its derivative according to claim 1 or 2, comprising a CDR3 domain having the sequence Antigen-binding fragment.

4. 1. An isolated antibody or antigen-binding fragment thereof that binds to human NAMPT, (a) a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 3, a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 4 and a CDR2 domain having the amino acid sequence shown in SEQ ID NO:

5. and a heavy chain variable region comprising a CDR3 domain having the amino acid sequence shown in SEQ ID NO:

6. a CDR1 domain having the amino acid sequence set forth in SEQ ID NO:7; and a CDR2 domain having the amino acid sequence set forth in SEQ ID NO:

7. and a light chain variable region comprising a CDR3 domain having the amino acid sequence set forth in SEQ ID NO:

8. 、 (b) a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 3, a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 4 and a CDR2 domain having the amino acid sequence shown in SEQ ID NO:

5. and a heavy chain variable region comprising a CDR3 domain having the amino acid sequence shown in SEQ ID NO:

11. a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 12; and a light chain variable domain comprising a CDR3 domain having the amino acid sequence set forth in SEQ ID NO:

8. region, (c) a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 3, a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 4 and a CDR2 domain having the amino acid sequence shown in SEQ ID NO:

5. and a heavy chain variable region comprising a CDR3 domain having the amino acid sequence shown in SEQ ID NO:

11. a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 14; and a light chain variable domain comprising a CDR3 domain having the amino acid sequence set forth in SEQ ID NO:

8. region, (d) a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 3, a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 29 and a CDR2 domain having the amino acid sequence shown in SEQ ID NO:

5. a heavy chain variable region comprising a CDR3 domain having the amino acid sequence shown in SEQ ID NO:11; a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 14; a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 15; a light chain containing a main CDR3 domain having the amino acid sequence shown in SEQ ID NO:8; variable domain, (e) a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 3, a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 29 and a CDR2 domain having the amino acid sequence shown in SEQ ID NO:

5. a heavy chain variable region comprising a CDR3 domain having the amino acid sequence shown in SEQ ID NO:11; a CDR1 domain having the amino acid sequence shown in SEQ ID NO:33; a CDR2 domain having the amino acid sequence shown in SEQ ID NO:34; a light chain containing a main CDR3 domain having the amino acid sequence shown in SEQ ID NO:8; variable domain, (f) a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 3, a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 29 and a CDR2 domain having the amino acid sequence shown in SEQ ID NO:

5. a heavy chain variable region comprising a CDR3 domain having the amino acid sequence shown in SEQ ID NO:11; a CDR1 domain having the amino acid sequence shown in SEQ ID NO:35; a CDR2 domain having the amino acid sequence shown in SEQ ID NO:36; a light chain containing a main CDR3 domain having the amino acid sequence shown in SEQ ID NO:8; variable domain, (g) a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 3, a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 29 and a CDR2 domain having the amino acid sequence shown in SEQ ID NO:

5. a heavy chain variable region comprising a CDR3 domain having the amino acid sequence shown in SEQ ID NO:11; a CDR1 domain having the amino acid sequence shown in SEQ ID NO:37; a CDR2 domain having the amino acid sequence shown in SEQ ID NO:38; a light chain containing a main CDR3 domain having the amino acid sequence shown in SEQ ID NO:8; variable domain, (h) a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 3, a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 4 and a CDR2 domain having the amino acid sequence shown in SEQ ID NO:

5. and a heavy chain variable region comprising a CDR3 domain having the amino acid sequence shown in SEQ ID NO:

11. a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 33; and a light chain variable domain comprising a CDR3 domain having the amino acid sequence set forth in SEQ ID NO:

8. region, (i) a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 3, a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 4 and a CDR2 domain having the amino acid sequence shown in SEQ ID NO:

5. and a heavy chain variable region comprising a CDR3 domain having the amino acid sequence shown in SEQ ID NO:

11. a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 35; and a light chain variable domain comprising a CDR3 domain having the amino acid sequence set forth in SEQ ID NO:

8. Area, or (j) a CDR1 domain having the amino acid sequence shown in SEQ ID NO: 3, a CDR2 domain having the amino acid sequence shown in SEQ ID NO: 4 and a CDR2 domain having the amino acid sequence shown in SEQ ID NO:

5. and a heavy chain variable region comprising a CDR3 domain having the amino acid sequence shown in SEQ ID NO:

11. a CDR1 domain having the amino acid sequence shown in SEQ ID NO:37; and a light chain variable domain comprising a CDR3 domain having the amino acid sequence set forth in SEQ ID NO:

8. An isolated antibody or antigen-binding fragment thereof comprising a region.

5. The isolated antibody or antigen-binding thereof according to any one of claims 1 to 4, which is humanized. Fragment.

6. The light chain variable region is set forth in SEQ ID NO: 2, 10, 13, 30, 31, 32, 34, or 36. The isolated antibody or its derivatives according to any one of claims 1 to 5, having an amino acid sequence Antigen-binding fragment of.

7. The heavy chain variable region has the amino acid sequence shown in SEQ ID NO: 1, 9, 15, 16 or 28. The isolated antibody or antigen-binding fragment thereof according to any one of claims 1 to 6, to.

8. Isolation of binding to human nicotinamide phosphoribosyltransferase (NAMPT) an antibody or antigen-binding fragment thereof having the amino acid sequence shown in SEQ ID NO: 15; a heavy chain comprising a variable region comprising the amino acid sequence shown in SEQ ID NO: 13; and a light chain comprising:

9. An isolated antibody or antigen-binding fragment thereof that binds to human NAMPT, comprising the sequence a heavy chain comprising a variable region comprising the amino acid sequence shown in SEQ ID NO: 26 and a heavy chain comprising a variable region comprising the amino acid sequence shown in SEQ ID NO: 18; and a light chain comprising a variable region comprising an amino acid sequence selected from the group consisting of: Fragment.

10. An isolated antibody or antigen-binding fragment thereof that binds to human NAMPT, comprising: heavy chains comprising a variable region as set forth in Table 17 and and a light chain comprising a nucleotide sequence ...

11. An isolated antibody or antigen-binding fragment thereof that binds to human NAMPT, comprising: a heavy chain comprising a heavy chain CDR1, CDR2 and CDR3 as set forth in Table 17; and a light chain comprising a light chain CDR1, CDR2, and CDR3 as described above. An antibody or antigen-binding fragment thereof.

12. An isolated anti-NAMPT antibody comprising the heavy chain variable region and the light chain variable region of antibody AL-303. body.

13. An isolated anti-NAMPT antibody comprising the heavy chain variable region and the light chain variable region of antibody AL-310. body.

14. A humanized heavy chain variable region derived from mouse antibody AL-303 or AL-310 and a mouse antibody An isolated human antibody comprising a humanized light chain variable region derived from antibody AL-303 or AL-310. anti-NAMPT antibody.

15. An isolated anti-NAMPT antibody that specifically binds to human NAMPT in a homodimeric structure. So, a) at least one amino acid of amino acid residues 17 to 44 of SEQ ID NO: 60, the sequence At least one amino acid among amino acid residues 117 to 127 of SEQ ID NO: 60, SEQ ID NO: 6 at least one amino acid of amino acid residues 162-170 of SEQ ID NO: 60; at least one amino acid of amino acid residues 242-261, amino acid of SEQ ID NO: 60 At least one amino acid of residues 262-273, amino acid residue 2 of SEQ ID NO:60 at least one amino acid of amino acid residues 332 to 305 of SEQ ID NO:60; at least one amino acid of 342, amino acid residues 374-389 of SEQ ID NO:60 at least one amino acid selected from amino acid residues 418 to 425 of SEQ ID NO: 60; at least one amino acid of at least one amino acid of amino acid residues 453 to 466 of SEQ ID NO: 60; at least one amino acid and at least one amino acid residue between 408 and 416 of SEQ ID NO:60 an epitope on human NAMPT comprising at least one amino acid; b) at least one amino acid of amino acid residues 29 to 51 of SEQ ID NO: 60, the sequence At least one amino acid of amino acid residues 61 to 72 of SEQ ID NO: 60 At least one amino acid of amino acid residues 156 to 170, amino acid of SEQ ID NO: 60 At least one amino acid of amino acid residues 216 to 234, amino acid residues of SEQ ID NO: 60 at least one amino acid of 316-331, amino acid residue 332 of SEQ ID NO:60 at least one amino acid of amino acid residues 373-38 of SEQ ID NO:60; 9, amino acid residues 417 to 431 of SEQ ID NO:60 at least one amino acid of at least one of amino acid residues 454 to 469 of SEQ ID NO:60 At least one amino acid and at least one amino acid residue between 470 and 478 of SEQ ID NO:60 Human NAMPT comprising any epitope on human NAMPT containing at least one amino acid. An isolated anti-NAMPT antibody that binds to an epitope on PT.

16. The isolated antibody or its derivatives according to any one of claims 1 to 15, comprising an Fc domain. Antigen-binding fragment.

17. The isolated antibody or antibody of any one of claims 1 to 16, which is a monoclonal antibody. is its antigen-binding fragment.

18. The isolated antibody or its antibody of any one of claims 1 to 17, which is an IgG antibody. Original binding fragment.

19. 19. The isolated antibody or its antigen binding site of claim 18, which is an IgG1 or IgG4 antibody. Combined fragment.

20. The isolated antibody or antigen-binding fragment thereof of any one of claims 1 to 19. A nucleic acid encoding

21. A vector comprising the nucleic acid of claim 20.

22. 22. A host cell comprising the nucleic acid of claim 20 or the vector of claim 21.

23. The isolated antibody or antigen-binding fragment thereof of any one of claims 1 to 19. A pharmaceutical composition comprising:

24. A method of treating a disorder associated with adverse NAMPT activity in a subject in need thereof. an effective amount of the isolated antibody or antigen-binding fragment of any one of claims 1 to 19 administering to said subject a compound.

25. 20. A method of treating a subject having an inflammatory condition comprising administering an effective amount of any of claims 1 to 19.

2. A method comprising administering to a subject the isolated antibody or antigen-binding fragment of claim 1. Law.

26. The inflammatory diseases include pulmonary fibrosis (IPF), pulmonary hypertension, acute lung injury (ALI), acute respiratory distress syndrome (SARS), and pulmonary fibrosis (PFS). respiratory distress syndrome (ARDS), ventilator-induced lung injury (VILI), ARDS / VILI Induced ALI, trauma-induced acute lung injury (TIALI) and brain injury or radiation-induced lung injury 26. The method of claim 25, wherein:

27. 2. The method of claim 1, wherein the radiation-induced lung injury is caused by radiation associated with cancer treatment.

26. The method according to claim 26.

28. A method of treating prostate cancer (PCa) in a subject in need thereof, comprising administering an effective amount of a compound according to claim 1.

20. The isolated antibody or antigen-binding fragment thereof according to any one of claims 1 to 19, administering to an elephant.

29. 29. The method of claim 28, wherein the subject has recurrent PCa.

30. 29. The method of claim 28, wherein the subject is at risk of developing metastatic PCa.

31. The PCa is resistant to androgen deprivation therapy (ADT).

10. The method according to any one of claims 1 to 9.

32. The method of any one of claims 28 to 31, further comprising administering ADT to the subject. How to do it.

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