Anti-MYCT1 antibodies and uses thereof

Abstract

In various aspects of the present disclosure, compositions comprising an anti-MYCT1 antibody and methods for modulating MYCT1 activity are provided. One aspect of the present disclosure provides a method of modulating tumor angiogenesis and / or anti-tumor immunity in a subject that inhibits tumor growth.

Description

[0001] Cross-references

[0002] This application claims the benefit of U.S. Provisional Patent Application No. 63 / 357,888, filed on July 1, 2023, entitled “ANTI-MYCT1 ANTIBODIES AN D USESTHEREOF,” the contents of which are hereby incorporated by reference in their entirety. Technical Field

[0003] The present disclosure generally relates to compositions comprising anti-MYCT1 antibodies and methods for modulating MYCT1 activity. More specifically, the present disclosure provides compositions and methods for preventing or reducing tumor angiogenesis, remodeling tumor immunity, and improving the outcome of immunotherapy.

[0004] Incorporation of Sequence Listing

[0005] This application contains a sequence listing filed in .XML format via the Patent Center, and is hereby incorporated by reference in its entirety. The WIPO sequence listing was created on July 3, 2023, the XML copy is named 760394.xml, and is 137 kilobytes in size. Background Art

[0006] Both angiogenesis and immune tolerance are normal physiological mechanisms that are hijacked by tumors. Angiogenesis involves the formation of new blood vessels from pre-existing vessels during development and wound healing. The regulation of angiogenesis is highly regulated by pro-angiogenic and anti-angiogenic factors, a process that becomes perturbed and dysregulated in cancer. Tumor-driven hypoxia increases the expression of pro-angiogenic factors, leading to the formation of new blood vessels that are critical for tumor survival and proliferation. The VEGF family, consisting of six growth factors (VEGFA-F), plays the most critical role in angiogenesis by binding to their receptors VEGFR1-3 and neuropilin. Angiogenesis can also be mediated by the angiopoietin (Ang1-2) / Tie-2 pathway, independent of the VEGF pathway. Therefore, drug development over the past decade has focused primarily on anti-angiogenesis as a strategy to deprive tumors of nutrients and inhibit tumor growth. However, despite the modest activity of these agents as single agents or in combination with chemotherapy, tumors may still overcome their effects and become resistant.

[0007] With the discovery of immune checkpoints, cancer immunotherapy has emerged as a way to effectively treat a variety of cancers. Many studies of immune checkpoint inhibitors (ICIs) have demonstrated long-term clinical activity against many malignancies. ICI blocks another mechanism hijacked by tumor exhaustion, "immune exhaustion," releasing effector immune cells against cancer. Primary resistance to ICIs has been described in tumors lacking tumor-infiltrating lymphocytes. In addition, tumors that initially respond to ICIs may develop secondary resistance due to defects in antigen presentation mechanisms and overexpression of co-inhibitory molecules, as well as other factors.

[0008] Therefore, there is a need in the art for compositions and methods that address angiogenesis and immune depletion in tumor cells. SUMMARY OF THE INVENTION

[0010] In some aspects, provided herein is an isolated anti-MYCT1 antibody or fragment thereof that specifically binds to an epitope within the first 100 amino acids of SEQ ID NO:145. In some aspects, the isolated anti-MYCT1 antibody or fragment thereof comprises a light chain variable region and / or a heavy chain variable region, wherein the light chain variable region comprises L1 having an amino acid sequence of SEQ ID NO: 28, SEQ ID NO: 64, SEQ ID NO: 100, or SEQ ID NO: 136, L2 having an amino acid sequence of SEQ ID NO: 30, SEQ ID NO: 66, SEQ ID NO: 102, or SEQ ID NO: 138, L3 having an amino acid sequence of SEQ ID NO: 32, SEQ ID NO: 68, SEQ ID NO: 104, or SEQ ID NO: 140, or any combination thereof; the heavy chain variable region comprises H1 having an amino acid sequence of SEQ ID NO: 10, SEQ ID NO: 46, SEQ ID NO: 82, or SEQ ID NO: 118, H2 having an amino acid sequence of SEQ ID NO: 12, SEQ ID NO: 48, SEQ ID NO: 84, or SEQ ID NO: 120, H3 having an amino acid sequence of SEQ ID NO: 14, SEQ ID NO: 50, SEQ ID NO: 51, SEQ ID NO: 52, SEQ ID NO: 53, SEQ ID NO: 54, SEQ ID NO: 55, SEQ ID NO: 56, SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 59, SEQ ID NO: 60 H3 of the amino acid sequence of SEQ ID NO:86 or SEQ ID NO:122, or any combination thereof.

[0011] In some aspects, the isolated anti-MYCT1 antibody or fragment thereof comprises a light chain variable region and / or a heavy chain variable region, wherein the light chain variable region comprises the amino acid sequence of SEQ ID NO:36, SEQ ID NO:72, SEQ ID NO:108 or SEQ ID NO:144; the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:18, SEQ ID NO:54, SEQ ID NO:90 or SEQ ID NO:126, or any combination thereof.

[0012] In some aspects, the isolated anti-MYCT1 antibody or fragment thereof comprises a light chain variable region and / or a heavy chain variable region, wherein the light chain variable region comprises L1 having an amino acid sequence of SEQ ID NO: 28, L2 having an amino acid sequence of SEQ ID NO: 30, L3 having an amino acid sequence of SEQ ID NO: 32, or any combination thereof; the heavy chain variable region comprises H1 having an amino acid sequence of SEQ ID NO: 10, H2 having an amino acid sequence of SEQ ID NO: 12, H3 having an amino acid sequence of SEQ ID NO: 14, or any combination thereof.

[0013] In some aspects, the isolated anti-MYCT1 antibody or fragment thereof comprises a light chain variable region and / or a heavy chain variable region, the light chain variable region comprising L1 having an amino acid sequence of SEQ ID NO: 64, L2 having an amino acid sequence of SEQ ID NO: 66, L3 having an amino acid sequence of SEQ ID NO: 68, or any combination thereof; the heavy chain variable region comprises H1 having an amino acid sequence of SEQ ID NO: 46, H2 having an amino acid sequence of SEQ ID NO: 48, H3 having an amino acid sequence of SEQ ID NO: 50, or any combination thereof.

[0014] In some aspects, the isolated anti-MYCT1 antibody or fragment thereof comprises a light chain variable region and / or a heavy chain variable region, the light chain variable region comprising L1 having the amino acid sequence of SEQ ID NO: 100, L2 having the amino acid sequence of SEQ ID NO: 102, L3 having the amino acid sequence of SEQ ID NO: 104, or any combination thereof; the heavy chain variable region comprises H1 having the amino acid sequence of SEQ ID NO: 82, H2 having the amino acid sequence of SEQ ID NO: 84, H3 having the amino acid sequence of SEQ ID NO: 86, or any combination thereof.

[0015] In some aspects, the isolated anti-MYCT1 antibody or fragment thereof comprises a light chain variable region and / or a heavy chain variable region, the light chain variable region comprising L1 having an amino acid sequence of SEQ ID NO: 136, L2 having an amino acid sequence of SEQ ID NO: 138, L3 having an amino acid sequence of SEQ ID NO: 140, or any combination thereof; the heavy chain variable region comprises H1 having an amino acid sequence of SEQ ID NO: 118, H2 having an amino acid sequence of SEQ ID NO: 120, H3 having an amino acid sequence of SEQ ID NO: 122, or any combination thereof.

[0016] In some aspects, the isolated anti-MYCT1 antibody or fragment thereof comprises the amino acid sequence of the light chain variable region of SEQ ID NO:36 and / or the amino acid sequence of the heavy chain variable region of SEQ ID NO:18.

[0017] In some aspects, the isolated anti-MYCT1 antibody or fragment thereof comprises the amino acid sequence of the light chain variable region of SEQ ID NO:72 and / or the amino acid sequence of the heavy chain variable region of SEQ ID NO:54.

[0018] In some aspects, the isolated anti-MYCT1 antibody or fragment thereof comprises the amino acid sequence of the light chain variable region of SEQ ID NO: 108 and / or the amino acid sequence of the heavy chain variable region of SEQ ID NO:90.

[0019] In some aspects, the isolated anti-MYCT1 antibody or fragment thereof comprises the amino acid sequence of the light chain variable region of SEQ ID NO: 144 and / or the amino acid sequence of the heavy chain variable region of SEQ ID NO: 126.

[0020] In some aspects, any of the isolated antibodies or fragments thereof disclosed herein are monoclonal antibodies. In some aspects, the isolated antibodies or fragments thereof are humanized antibodies, single chain variable fragments (scFv) antibodies, antibody fragments or chimeric antibodies.

[0021] In some aspects, a pharmaceutical composition is further provided, comprising a disclosed isolated antibody or fragment thereof and a pharmaceutically acceptable carrier or excipient. In some aspects, the isolated antibody is used as a medicament.

[0022] The present disclosure further encompasses a method of treating cancer in a subject in need thereof, the method comprising administering an effective amount of a composition comprising an isolated antibody or fragment thereof disclosed herein. In some aspects, the method further comprises administering an immune checkpoint inhibitor and an optional anti-VEGF targeted therapy.

[0023] In other aspects, the present disclosure encompasses methods for sensitizing cancer in a subject to immune checkpoint inhibitors, the methods comprising: selecting a subject with cancer, wherein the cancer is resistant to immune checkpoint inhibitors; and administering to the subject an effective amount of a composition comprising a disclosed isolated antibody or fragment thereof. In some aspects of the method, administration of an anti-MYCT1 antibody causes a decrease in angiogenesis in cancer cells of the subject. In some aspects of the method, administration of an anti-MYCT1 antibody causes an enhancement of anti-tumor immunity in the subject. In some aspects of the method, administration of an anti-MYCT1 antibody causes a decrease in anti-tumor volume in the subject.

[0024] Further provided is a method for regulating tumor angiogenesis and / or enhancing anti-tumor immunity in a subject with cancer, the method comprising: administering to the subject a therapeutically effective amount of a composition comprising an isolated antibody or fragment thereof disclosed herein. In some aspects, the method further comprises administering an immune checkpoint inhibitor and an optional VEGF inhibitor. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] Those skilled in the art will appreciate that the drawings described below are for illustration purposes only and are not intended to limit the scope of the present teachings in any way.

[0026] Figures 1A-1D The generation of anti-MYCT1 antibodies is shown. Figure 1A Shown is a sequence alignment of human MYCT1 isoform 1 (SEQ ID NO: 145) and mouse MYCT1 amino acid sequence (SEQ ID NO: 150). Figure 1B The immunization scheme for generating MYCT1 monoclonal antibodies is shown. LS, dioxetine synthase (LS) from Aquifex aeolicus (AaLS), keyhole limpet haemocy anin (KLH), bovine serum albumin (BSA), HUVEC, human umbilical vein endothelial cells (HUVEC), mouse cardiac endothelial cells (MCEC), enzyme-linked immunosorbent assay (ELISA), fluorescence-activated cell sorting (FACS), KD, knockdown (KD), overexpression (OE). Figure 1C Graph showing a representative MYCT1 terminal sera titration from fusion number 4941 (animal number 8). MYCT1 antigen was added at 1 μg / ml. Figure 1D Graph showing a representative MYCT1 terminal serum titration from fusion number 4952 (animal number 2). MYCT1 antigen was added at 1 μg / ml.

[0027] Figure 2A-2E Plasmid maps of plasmids used in the studies disclosed herein are shown.

[0028] Figures 3A-3D Preliminary screening results of the generated monoclonal antibodies are shown. Figure 3A Results from ELISA assays are shown. Figure 3B Shown are results from fluorescence activated cell sorting (FACS), mouse cardiac endothelial cells (MCEC), overexpression (OE). Figure 3C and Figure 3D Representative results and images from the scratch assay at 3 hours are shown.

[0029] Figures 4A-4C Display usage Figure 2A Screening results for plasmid pcDNA3.1-Myct1Flag are shown. Figure 4A Results from FACS are shown. Figure 4B and Figure 4C Representative image results from the scratch assay are shown.

[0030] Figure 5A-5B Characterization of anti-MYCT1 antibody clones is shown. Figure 5A Shown are the identification of clones 2B4 (IgG2b isotype) and 11B6 (IgG1 isotype) based on scratch assay, FACS staining and ELISA. Figure 5B The identification of clones 3E3, 8D12, 9E2 and 9F8 based on scratch assay, FACS staining and ELISA is shown.

[0031] Figures 6A-6C Characterization of antibody clone 2B4F9D9 is shown. Fig. 6A Shown is FACS using hybridoma supernatants containing candidate antibody 2B4F9D9 in HA-mouse Myct1 overexpressing (HA-mMOE) murine cardiac endothelial cells (MCECs). Figure 6B and Figure 6C Representative images and results from a scratch assay using antibody clone 2B4F9D9 are shown.

[0032] Figures 7A-7C Shown are results from a sprouting assay in induced murine lung endothelial cells (iMLEC) treated with purified antibody clones for 15 hours. Fig. 7A Shown are representative images of budding assays in iMLEC cells and iMLEC Mytc1 KO cells. Figure 7B Shown are buds / beads in iMLEC cells and iMLEC Mytc1 KO cells. Figure 7CShown are the sprout lengths in iMLEC cells and iMLEC Mytc1 KO cells. **p<0.01, ***p<0.001, ****p<0.0001 compared to IgG-treated cells, one-way ANOVA.

[0033] Figures 8A-8C Shown are results from a budding assay in MCECs treated with purified antibody clones for 30 hours. Fig. 8A Shown are representative images of the sprouting assay in MCEC cells. Figure 8B Shown are buds / beads in MCE C cells. Figure 8C Shown are sprout lengths in MCEC cells. **p<0.01 compared to IgG-treated cells, one-way ANOVA.

[0034] Figures 9A-9G Shown are results from sprouting assays in MCEC cells or induced human umbilical vein endothelial cell (iHUVEC) cells treated with purified antibody clones 3EC12, 8D2C12, 9E2H5, and 2B4F9D3. Fig.9A Shown are representative images of the sprouting assay in MCEC cells. Fig. 9B Shown are sprouts / beads in MCEC cells, *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001 compared to HM-treated group, one-way ANOVA. Fig. 9C Shown are sprout lengths in MCEC cells. ***p<0.001, ****p<0.0001 compared to HM-treated group, one-way ANOVA. Fig.9D Shown are representative images of budding assays in iHUVEC cells treated for 24 hours with increasing concentrations of antibody clones 11B6 or 2B4F9D3. Fig.9E Shown are representative images of budding assays in iHU VEC cells treated for 48 hours with increasing concentrations of antibody clones 11B6 or 2B4F9D3. Fig.9F Graph showing sprout number and sprout length in iHUVEC cells treated for 24 hours with increasing concentrations of antibody clones 11B6 or 2B4F9D3. *p<0.05 compared to IgG treated cells, two-way ANOVA. Figure 9G Graph showing sprout number and sprout length in iHUVEC cells treated for 48 hours with increasing concentrations of antibody clones 11B6 or 2B4F9D3. *p<0.05, ***p<0.001, ****p<0.0001 compared to IgG treated cells, two-way ANOVA.

[0035] Figures 10A-10EResults from a tube formation assay with purified antibody clones are shown. Fig. 10A Representative images showing tube formation in iMLEC cells treated with 4 μg / well of 11B6 and 2B4F9D3 for 4 hours. Fig. 10B Representative images showing tube formation in MCEC cells treated with 2B4F9D3, 3EC12, 8D2C12, and 9E2H5. Fig. 10C The number of rings in each treatment in MCECs is shown. **p<0.01, ***p<0.001 compared to HM-treated cells, one-way ANOVA. Fig. 10D Tube formation in iHUVEC cells treated with increasing concentrations of 11B6 or 2B4F9D3 for 16 hours is shown. Scale bar = 100 μm. Fig. 10E The number of loops and branches in treated iHUVEC cells is shown. *p<0.05, **p<0.01 compared to IgG-treated cells, two-way ANOVA.

[0036] Figures 11A-11F Shown are the results from a scratch assay of cells treated with hybridoma supernatants. Fig.11A Representative images of wound closure at 0, 8 and 10 hours in MCEC cells treated with hybridoma supernatants of antibody clones 11B6, 2B4F9D3 or IgG added at 32 μg / well are shown. Fig. 11B Shown are graphs depicting relative closed tubes in MCEC cells treated with hybridoma supernatants of 11B6, 2B4F9D3 or IgG. *p<0.05, **p<0.01, two-way ANOVA compared to IgG treated cells. Fig. 11C Representative images of wound closure at 0, 9, and 11 hours in iMLEC cells and iMLEC Myct1 knockout (MKO) cells treated with hybridoma supernatants of 11B6, 2B4F9D3, or IgG added at 32 μg / well are shown. Fig.11D Shown are graphs depicting relative closed tubes in iMLEC cells and iMLEC MKO cells treated with hybridoma supernatants of 11B6, 2B4F9D3 or IgG. *p<0.05, **p<0.01 compared to IgG treated cells, two-way ANOVA. Fig.11E Representative images of wound closure at 0 and 9 hours in MCEC cells treated with hybridoma supernatants of 3E3C12, 8D2C12, 9E2H5, 2B4F9D3, or IgG added at 50 μg / well are shown. Fig.11FShown are graphs depicting relative closed tubes in MCEC cells treated with hybridoma supernatants of 3E3C12, 8D2C12, 9E2H5, 2B4F9D3, or HM. **p<0.01, ****p<0.0001 compared to HM-treated cells, one-way ANOVA.

[0037] Figures 12A-12F In vivo characterization of 2B4F9D3 and 11B6 using tumor models is shown. Fig. 12A and Fig. 12C A regimen of 11B6 or 2B4F9D3 injections with α-PD1 antibody treatment using the 1956 sarcoma line is shown. Fig. 12B and Fig.12D Shown are tumor growth in PBS-injected (control), α-PD1 alone, 2B4F9D3+α-PD1, or 11B6+α-PD1 treated mice. Fig.12E Shown is the regimen of 11B6 or 2B4F9D3 injection with α-PD1 antibody treatment using the PyMT-BO1 orthotopic breast tumor model. Fig.12F Shown are tumor growth in PBS-injected (control), α-PD1 alone, 2B4F9D3+α-PD1, or 11B6+α-PD1 treated mice. Figure 12G Merged line graph showing tumor growth. α-PD1, 250 μg / injection / mouse, 2B4 and 11B6, 330 μg / injection / mouse. All antibodies were injected intraperitoneally. Summary of the invention

[0038] Both immune checkpoint inhibitors (ICI) and anti-angiogenic agents have changed the landscape of modern cancer treatment. Although anti-angiogenic agents have demonstrated activity in highly vascularized tumors, including renal cell carcinoma and colorectal cancer, the effects of ICI are mainly observed in tumors with immune recognition of highly immune-infiltrated lymphocytes. The main challenge in ICI drug development is to migrate their activity to non-inflamed tumors and overcome drug resistance driven in part by immunosuppressive microenvironment. Angiogenic factors drive immunosuppression by directly inhibiting antigen presenting cells and immune effector cells or by enhancing the effects of regulatory T cells (Treg), myeloid-derived suppressor cells (MDSC) and tumor-associated macrophages (TAM). Those suppressive immune cells can also drive angiogenesis, resulting in a vicious cycle of impaired immune activation. The applicant found that compositions and certain methods comprising MYCT 1 reduce or prevent tumor growth, reduce or prevent tumor angiogenesis (i.e., the formation and development of the vascular system required by the tumor in order to thrive and progress) and / or increase tumor immune response.

[0039] MYCT1 protein does not appear to be required for steady-state angiogenic signaling, but is essential for maintaining an immunosuppressive tumor vasculature and tumor microenvironment (Kabir et al., Sci. Transl. Med. 13, eab b6731 (2021), the contents of which are incorporated herein by reference). The present disclosure is based in part on the identification of anti-MYCT1 monoclonal antibodies that specifically bind to epitopes within the extracellular region of MY CT1 protein. MYCT1 protein exists in two isoforms in humans. One isoform has a sequence of 235 amino acids (referred to herein as isoform 1), and a shorter second isoform has 187 amino acids (referred to herein as isoform 2). Amino acids 1-19 of mouse MYCT1 are identical to amino acids 48-67 of human MYCT1 isoform 1 ( Figure 1A ) and amino acids 1-19 of human MYCT1 isoform 2 are the same. This conserved region targeting MYCT1 is used to generate the antibodies herein. The disclosed antibodies generated using this extracellular region of MYCT1 have a wide range of applicability, including neutralizing activity against both human MYCT1 protein and mouse MYCT1 protein. Therefore, the present disclosure encompasses the use of compositions and methods comprising anti-MYCT1 antibodies for treating tumors or cancers in subjects. In particular, the present disclosure provides for controlling tumor angiogenesis and reprogramming tumor immunity requiring MYCT1 expression and / or activity. Further provided herein are compositions and methods for downregulating MYCT1 activity and / or expression in cells (e.g., endothelial cells). Downregulation of MYCT1 is associated with reducing angiogenesis, enhancing endothelial venule formation, and promoting an anti-tumor immune environment, resulting in limited tumor progression. Therefore, the present disclosure provides compositions and methods for reducing or preventing angiogenesis and increasing tumor immune responses.

[0040] Other aspects and reiterations of the invention are more fully described below.

[0041] I. Terminology

[0042] In order to more easily understand the present invention, certain terms are defined. Unless otherwise defined, all technical and scientific terms used herein have the same meanings as those of ordinary skill in the art generally understood with respect to aspects of the present invention. Many methods and materials similar to those described herein, modified or equivalent can be used in the practice of aspects of the present invention without excessive experimentation, and preferred materials and methods are described herein. When describing and claiming aspects of the present invention, the following terms will be used according to the definition set forth below.

[0043] The term "a" or "an" entity refers to one or more of the entity; for example, "polypeptide subunit" is understood to represent one or more polypeptide subunits. Therefore, the terms "a" (or "an"), "one or more", and "at least one" are used interchangeably herein.

[0044] As used herein, the term "about" refers to the variation in numerical quantity that can occur, for example, by typical measuring techniques and equipment, about any quantifiable variable, including but not limited to mass, volume, time, distance, and amount. In addition, given solid and liquid handling procedures used in the real world may have certain unintentional errors and variations due to differences in the manufacture, source, or purity of the ingredients used to prepare the composition or implement the method, etc. The term "about" also covers these variations, which can be up to ±5%, but can also be ±4%, 3%, 2%, 1%, etc. Whether or not modified by the term "about", the claims include equivalents to the quantities.

[0045] The phrase "and / or" as used herein in the specification and claims should be understood to mean "either or both" of the elements so combined, i.e., elements that are present in combination in some cases and separately in other cases. Multiple elements listed with "and / or" should be interpreted in the same manner, i.e., "one or more" of the elements so combined. Other elements other than the elements specifically indicated by the "and / or" clause may optionally be present, whether related or unrelated to those elements specifically indicated. Thus, as a non-limiting example, when used in conjunction with open language such as "comprising", references to "A and / or B" may, in one aspect, refer only to A (optionally including elements other than B); in another aspect, refer only to B (optionally including elements other than A); in yet another aspect, refer to both A and B (optionally including other elements); and so on.

[0046] As used herein in the specification and claims, "or" should be understood to have the same meaning as "and / or" as defined above. For example, when separating items in a list, "or" or "and / or" should be interpreted as inclusive, i.e., including at least one of a plurality of elements or lists of elements, but also including more than one of a plurality of elements or lists of elements, and optional additional unlisted items. Only in terms with clear opposite indications, such as "only one of ..." or "exactly one of ...", or when used in the claims, "consisting of ...", will refer to including exactly one element in a plurality of elements or lists of elements. In general, when before an exclusive term, the term "or" as used herein should only be interpreted as indicating exclusive alternatives (i.e., "one or the other, but not both"), such as "either", "one of ...", "only one of ..." or "exactly one of ...". When used in the claims, "consisting essentially of ..." should have its ordinary meaning as used in the field of patent law.

[0047] As used herein in the specification and claims, the phrase "at least one" referring to a list of one or more elements should be understood to mean at least one element selected from any one or more elements in the list of elements, but does not necessarily include at least one of each and every element specifically listed in the list of elements, and does not exclude any combination of elements in the list of elements. This definition also allows that elements other than the elements specifically indicated in the list of elements referred to by the phrase "at least one" may optionally be present, whether related or unrelated to those elements specifically indicated. Thus, as a non-limiting example, "at least one of A and B" (or equivalently, "at least one of A or B" or equivalently "at least one of A and / or B") may, in one aspect, refer to at least one A, optionally including more than one A, wherein B is absent (and optionally including elements other than B); in another aspect, refer to at least one B, optionally including more than one B, wherein A is absent (and optionally including elements other than A); in yet another aspect, refer to at least one A, optionally including more than one A, and at least one B, optionally including more than one B (and optionally including other elements); and so on.

[0048] Unless otherwise defined, all technical terms used herein have the same meanings as those of ordinary skill in the art to which the present invention belongs. As used in this specification and the appended claims, the singular forms "a kind of", "one" and "said / the" include plural references unless the context clearly indicates otherwise. Unless otherwise indicated, any reference to "or" herein is intended to cover "and / or". Whenever the term "at least", "greater than" or "greater than or equal to" is before the first value in a series of two or more numerical values, the term "at least", "greater than" or "greater than or equal to" is applicable to each numerical value in the series of numerical values. For example, greater than or equal to 1, 2 or 3 is equivalent to greater than or equal to 1, greater than or equal to 2 or greater than or equal to 3.

[0049] Whenever the term "not more than," "less than," "less than or equal to," or "at most" precedes the first value in a series of two or more values, the term "not more than," "less than," "less than or equal to," or "at most" applies to each value in the series. For example, less than or equal to 3, 2, or 1 is equivalent to less than or equal to 3, less than or equal to 2, or less than or equal to 1.

[0050] Where values ​​are described as ranges, it should be understood that such disclosure includes disclosure of all possible sub-ranges within such ranges, as well as specific values ​​falling within such ranges, whether or not a specific value or a specific sub-range is explicitly stated.

[0051] Components to be used to prepare the disclosed compositions and the compositions themselves to be used in the methods disclosed herein. These and other materials are disclosed herein, and it should be understood that when the combinations, subsets, interactions, groups, etc. of these materials are disclosed, although the specific references of the various individual and collective combinations and arrangements of these compounds may not be explicitly disclosed, each is specifically considered and described herein. For example, if a specific compound is disclosed and discussed, and many modifications that can be made to many molecules of the compound are discussed, then unless otherwise specifically indicated, each and every combination and arrangement of the compound and possible modifications are specifically considered. Therefore, if a class of molecules A, B and C and a class of molecules D, E and F, as well as examples of combined molecules AD are disclosed, then even if each is not individually listed, each is considered individually and collectively, meaning that it is considered that the combination AE, AF, BD, BE, BF, CD, CE and CF are disclosed. Similarly, any subset or combination of these is also disclosed. Therefore, for example, it is considered that the subgroups of AE, BF and CE are disclosed. This concept applies to all aspects of the application, including but not limited to the steps of the method for preparing and using the disclosed composition. Thus, if there are a variety of additional steps that can be performed it is understood that each of these additional steps can be performed with any specific aspect or combination of aspects of the disclosed methods.

[0052] As used herein, the term "non-naturally occurring" matter, composition, entity, and / or any combination of matter, composition or entity, or any grammatical variation thereof, is a conditional term that specifically excludes, but only excludes, those forms of matter, composition, entity, and / or any combination of matter, composition or entity that are fully understood by one of ordinary skill in the art to be "naturally occurring" or that are at any time determined or interpreted by a judge or administrative or judicial body to be "naturally occurring" or that could be determined or interpreted by a judge or administrative or judicial body to be "naturally occurring."

[0053] "Specific binding" means that a binding molecule, such as an antibody or antigen binding fragment thereof, binds to an epitope via its antigen binding domain, and that the binding requires some recognition between the antigen binding domain and the epitope. According to this definition, a binding molecule is said to "specifically bind" to an epitope when it binds to the epitope more easily via its antigen binding domain than it would to a random, unrelated epitope.

[0054] The portion of a target polypeptide that specifically interacts with the antigen binding domain of an antibody is an "epitope". In addition, it should be noted that an "epitope" on MYCT1 can be a linear epitope or a conformational epitope, and in both cases can include non-polypeptide elements, for example, the epitope can include carbohydrate or lipid side chains. The term "affinity" refers to a measure of the strength of binding of an individual epitope to the antigen binding site of an antibody.

[0055] As used herein, the term "antibody" is used in the broadest sense and encompasses a variety of antibodies and antibody-like structures, including but not limited to full-length monoclonal, polyclonal and multispecific (e.g., bispecific, trispecific, etc.) antibodies, as well as heavy chain antibodies and antibody fragments, provided that they exhibit the desired antigen-binding activity. The domains of antibodies involved in binding to antigens are called "variable regions" or "variable domains," and are described in further detail below. A single variable domain may be sufficient to confer antigen-binding specificity. Preferably, but not necessarily, recombinantly produced antibodies useful for discovery. Antibodies may be glycosylated or may be non-glycosylated, although glycosylated antibodies may be preferred. "Isolated" antibodies are antibodies separated from components of their natural environment. In some aspects, the antibodies are purified to a purity greater than 95% or 99%, as determined by methods known in the art.

[0056] In addition to the antibodies described herein, it is possible to design antibody mimics or aptamers that function substantially the same as the antibodies of the present invention using methods known in the art." Antibody mimics " refers to a polypeptide or protein that can specifically bind to an antigen but is structurally unrelated to an antibody. Antibody mimics have a mass of about 3kDa to about 20kDa. Non-limiting examples of antibody mimics are affibody molecules, affilin, affimer, alphabody, anticalin, avimer, designed ankyrin repeat proteins (DARPin) and monoclonal antibodies. Aptamers are small nucleic acid ligands composed of RNA or single-stranded DNA oligonucleotides, and have high specificity and affinity to their targets. Aptamers interact and bind to their targets through structural recognition, a process similar to antigen-antibody reaction. Aptamers have a molecular weight lower than that of an antibody, typically about 8-25kDa.

[0057] The terms "full-length antibody" and "intact antibody" may be used interchangeably and refer to an antibody having a structure substantially similar to a natural antibody structure or having a heavy chain containing an Fc region as defined herein. The basic structural unit of a natural antibody comprises a tetramer. Each tetramer consists of two pairs of identical polypeptide chains, each pair having a "light" chain (about 25 kDa) and a "heavy" chain (about 50-70 kDa). Light chains are classified as γ, μ, α, and λ. Heavy chains are classified as γ, μ, α, δ, or ε, and the isotype of the antibody is defined as IgG, IgM, IgA, IgD, and IgE, respectively. The amino terminal portion of each light and heavy chain includes a variable region (VL and VH, respectively) of about 100-110 or more amino acid sequences that are primarily responsible for antigen recognition. The carboxyl terminal portion of each chain defines a constant region that is primarily responsible for effector function. Within the light and heavy chains, the variable and constant regions are connected by a "J" region of about 12 or more amino acids, with the heavy chain also including a "D" region of about 10 more amino acids. Intact antibodies are appropriately cross-linked via disulfide bonds, as known in the art.

[0058] The variable domains of the heavy and light chains of antibodies generally have similar structures, wherein each domain comprises four conserved framework regions (FRs) and three hypervariable regions (HVRs). (See, e.g., Kindt et al., Kuby Immunol ogy, 6th edition, WH Freeman and Co., p. 91 (2007).) A single VH or VL domain may be sufficient to confer antigen binding specificity. In addition, antibodies that bind to a specific antigen can be isolated using a VH or VL domain from an antibody that binds to the antigen to screen a library of complementary VL or VH domains, respectively. See, e.g., Portolano et al., J. Immunol. 150: 880-887 (1993); Clarkson et al., Nature 352: 624-628 (1991).

[0059] "Framework region" or "FR" refers to the variable domain residues other than the hypervariable region (HVR) residues. The FR of the variable domain is generally composed of four FR domains: FR1, FR2, FR3, and FR4. Thus, HVR and FR sequences generally appear in the following sequence: FR1-HVR1-FR2-HVR2-FR3-HVR3-FR4. As is known in the art, the FR domains of the heavy chain and light chain can be different.

[0060] As used herein, the term "hypervariable region" or "HVR" refers to each region of a variable domain that is hypervariable in sequence (also often referred to as a "complementarity determining region" or "CDR") and / or forms structurally defined loops ("hypervariable loops") and / or contains antigen contact residues ("antigen contacts"). In general, antibodies comprise six HVRs: three in VH (H1, H2, H3) and three in VL (L1, L2, L3). As used herein, "HVR derived from a variable region" refers to an HVR that has no more than two amino acid substitutions compared to the corresponding HVR from the original variable region. Exemplary HVRs herein include: (a) hypervariable loops occurring at amino acid residues 26-32 (L1), 50-52 (L2), 91-96 (L3), 26-32 (H1), 53-55 (H2), and 96-101 (H3) (Chothia and Lesk, J. Mol. Biol. 196:901-917 (1987)); (b) CDRs occurring at amino acid residues 24-34 (L1), 50-56 (L2), 89-97 (L3), 31-35b (H1), 50-65 (H2), and 95-102 (H3) (Kabat et al., Sequences of Proteins of Immunological Interest, 5th ed., Public Health Service, National Institutes of Health alth, Bethesda, MD (1991)); (c) antigenic contacts occurring at amino acid residues 27c-36 (L1), 46-55 (L2), 89-96 (L3), 30-35b (H1), 47-58 (H2), and 93-101 (H3) (MacCallum et al., J. Mol. Biol.262:732-745 (1996)); (d) CDR1-IMGT (positions 27-38), CDR2-IMGT (positions 56-65), and CDR3-IMGT regions (positions 105-116 or 105-117) based on IMGT unique numbering (Lefranc, "The IMGT unique numbering for Immunoglobulins, T cell receptors and Ig-like domains," The Immunologist, 1999, 7:132-136; Lefranc et al., Nucleic Acids Research, 2009, 37(Database Issue):D1006-D1012; Ehrenmann et al., "Chapter 2: Standardized Sequence and Structure Analysis of Antibody Using IMGT", Antibody Engineering, Volume 2, edited by Roland E. Kontermann and Stefan Dubel, 2010, Spring er-Verlag Berlin Heidelberg, doi:10.1007 / 978-3-642-01147-4; imgt.org / IMGTScientificChart / Nomenclature / IMGT-FRCDRdefinition), and (e) combinations of (a), (b), (c) and / or (d), as defined below for various antibodies of the present disclosure. Unless otherwise indicated, HVR residues and other residues (e.g., FR residues) in the variable domain that are assigned sequence identification numbers are numbered based on the IMGT unique numbering, see above. .

[0061] The term "Fc region" herein is used to define the C-terminal region of an immunoglobulin heavy chain containing at least a portion of a constant region. The term includes native sequence Fc regions and variant Fc regions. In one aspect, the human IgG heavy chain Fc region extends from Cys226 or from Pro230 to the carboxyl terminus of the heavy chain. However, the C-terminal lysine (Lys447) in the Fc region may or may not be present. Unless otherwise specified herein, the numbering of amino acid residues in the Fc region or constant region is according to the EU numbering system, also referred to as the EU index, as described in Kabat et al., Sequences of Proteins of Immunological Interest, 5th edition, Public Health Service, National Institutes of Health, Bethesda, Md. (1991).

[0062] A "variant Fc region" comprises an amino acid sequence that may differ from the amino acid sequence of a native Fc region due to one or more amino acid substitutions and / or due to a modified glycosylation pattern compared to a native Fc region or to an Fc region of a parent polypeptide. In one example, a variant Fc region may have from about one to about ten amino acid substitutions or from about one to about five amino acid substitutions in a native sequence Fc region or in an Fc region of a parent polypeptide. The variant Fc region herein may have at least about 80% homology, at least about 90% homology, or at least about 95% homology with a native sequence Fc region and / or with an Fc region of a parent polypeptide.

[0063] "Antibody fragment" refers to a molecule other than an intact antibody that contains a portion of an intact antibody that binds to the antigen to which the intact antibody binds. Non-limiting examples of antibody fragments include, but are not limited to, Fv, Fab, Fab', Fab'-SH, F(ab')2; single-chain forms of antibodies and higher-order variants thereof; single-domain antibodies and multispecific antibodies formed from antibody fragments.

[0064] Single chain forms of antibodies and their higher order forms may include, but are not limited to, single domain antibodies, single chain variable fragments (scFv), divalent scFv (di-scFv), trivalent scFv (tri-scFv), tetravalent scFv (tetra-sc Fv), diabodies and triabodies and tetrabodies. ScFv comprises a heavy chain and a light chain variable region connected by a joint. In most cases, but not all cases, the joint can be a peptide. The joint peptide is preferably about 5-30 amino acids in length or about 10-25 amino acids in length. Typically, the joint allows for stable variable domains without interfering with the correct folding and generation of active binding sites. In preferred aspects, the joint peptide is rich in glycine and serine or threonine. ScFv can be used to promote phage display or can be used for flow cytometry, immunohistochemistry or as a targeting domain. Methods for preparing and using scFv are known in the art. ScFv can also be conjugated to human constant domains (e.g., heavy chain constant domains derived from IgG domains, such as IgG1, IgG2, IgG3, or IgG4, or heavy chain constant domains derived from IgA, IgM, or IgE). Diabodies, triabodies, and tetrabodies, as well as higher-order variants, are typically produced by changing the length of the linker peptide from zero to several amino acids. Alternatively, it is also known in the art that multivalent binding antibody variants can be generated using self-assembly units connected to variable domains.

[0065] "Single domain antibody" refers to an antibody fragment composed of a single monomeric variable antibody domain.

[0066] Multispecific antibodies include bispecific antibodies, trispecific antibodies or antibodies with four or more specificities. Multispecific antibodies can be produced by combining the heavy and light chains of one antibody with the heavy and light chains of one or more other antibodies. These chains can be covalently linked.

[0067] The antibody disclosed herein can be a dual-affinity re-targeting antibody (DART). The DART format is based on the double antibody format of the homologous variable domains of the heavy chain and light chain of 2 kinds of antigen binding specificities separated on 2 separate polypeptide chains. Although the 2 polypeptide chains are non-covalently associated in the double antibody format, the DART format provides additional stability through the C-terminal disulfide bridge. DART can be produced in high quantity and quality and shows excellent stability in both formulation buffer and human serum.

[0068] As used herein, "monoclonal antibody" refers to an antibody derived from a single copy or clone, including, for example, any eukaryotic, prokaryotic or phage clone. "Monoclonal antibody" is not limited to antibodies produced by hybridoma technology. Monoclonal antibodies can be produced using hybridoma technology well known in the art, as well as recombinant technology, phage display technology, synthetic technology, or a combination of such technology and other technologies readily known in the art. In addition, the monoclonal antibody can be labeled with a detectable marker, fixed on a solid phase, and / or conjugated to a heterologous compound (e.g., an enzyme or toxin) according to methods known in the art.

[0069] As used herein, "heavy chain antibody" refers to an antibody composed of two heavy chains. The heavy chain antibody may be an IgG-like antibody from camel, llama, alpaca, shark, etc., or an IgNAR from cartilaginous fish.

[0070] As used herein, "humanized antibody" refers to a non-human antibody that has been modified to reduce the risk of non-human antibodies inducing an immune response in humans after administration, but retains binding specificity and affinity similar to the starting non-human antibody. Humanized antibodies bind to the same or similar epitopes as non-human antibodies. The term "humanized antibody" includes antibodies that are partially or completely composed of amino acid sequences derived from human antibody germlines by changing the sequence of antibodies with non-human hypervariable regions ("HVRs"). The simplest such changes can simply consist of replacing the mouse constant region with the constant region of a human antibody, thereby producing a human / mouse chimera that can have low immunogenicity that is acceptable for pharmaceutical use. Preferably, the variable region of the antibody is also humanized by techniques currently known in the art. For example, the framework region of the variable region can be replaced by the corresponding human framework region while retaining one, several or all six non-human HVRs. Some framework residues can be replaced with corresponding residues from non-human VL domains or VH domains (e.g., non-human antibodies from which HVR residues are derived), for example, to restore or improve the specificity or affinity of the humanized antibody. The substantially human framework region has at least about 75% homology (i.e., at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, or at least about 99% sequence identity) with a known human framework region sequence. HVRs may also be randomly mutated so that binding activity and affinity for antigens are maintained or enhanced in the context of a fully human germline framework region or a substantially human framework region. As mentioned above, it is sufficient to use an antibody fragment for the method of this discovery. In addition, as used herein, the term "humanized antibody" refers to an antibody comprising a substantially human framework region, at least one HVR from a non-human antibody, and any constant region present therein is a substantially human constant region. Substantially human constant regions have at least about 90% (i.e., about 90%, about 95%, or about 99% sequence identity) with a known human constant region sequence. Therefore, all parts of a humanized antibody, except possibly HVR, are substantially identical to corresponding pairs of one or more germline human immunoglobulin sequences.

[0071] If desired, the design of humanized immunoglobulin can be carried out as follows or using similar methods familiar to those skilled in the art (e.g., see Almagro et al., Front.Biosci.2008, 13 (5): 1619-33). The mouse antibody variable region is aligned with the most similar human germline sequence (e.g., by using BLAST or similar algorithms). The CDR residues from the mouse antibody sequence are transplanted into similar human "receptor" germlines. Subsequently, one or more positions (e.g., Vernier positions) near or within the framework of the CDR can be restored to the original mouse amino acids to achieve humanized antibodies with similar binding affinity to the original mouse antibody. Typically, several versions of humanized antibodies with different back mutations are generated and tested for activity empirically. The humanized antibody variant with the most similar characteristics to the parent mouse antibody and the least mouse framework return is selected as the final humanized antibody candidate.

[0072] As used herein with respect to an epitope binding agent, the term "specifically binds" means that the epitope binding agent generally does not cross-react to a significant extent with other epitopes on the protein of interest (eg, MYCT1) or on other proteins.

[0073] The terms "Myc target 1", "MYC target protein 1", "MTLC", "Myc target protein 1 in myeloid cells" or "MYCT1" encompass all MYCT isoforms and orthologs, whether full-length, truncated or post-translationally modified. In many animals, including but not limited to humans, non-human primates, rodents, fish, cattle, frogs, goats and chickens, MYCT1 is encoded by the gene Myct1 gene (also known as FLJ21269 and MT LC). The gene encoding MYCT1 is located on chromosome 6 in humans (band q25.2; chromosome location (bp) 152697895-152724567). MYCT1 was originally identified as a novel target of the c-Myc oncogene in myeloid cells. Previous reports suggest that overexpression in cancer cells can promote apoptosis, morphological changes, enhancement of anchorage-independent cell growth, tumorigenic transformation, promotion of genomic instability and inhibition of hematopoietic differentiation. MYCT1 was originally thought to be a transcription factor and binds to the promoters of several c-Myc-regulated genes, and it has been suggested that the phenotypes seen in MYCT1-overexpressing cells are a result of dysregulation of these genes. In humans, the MYCT1 protein has two isoforms, isoform 1 with a sequence of 235 amino acids in length and a shorter isoform 2 with a sequence of 187 amino acids in length. Amino acids 1-19 of mouse MYCT1 are identical to amino acids 48-67 of human MYCT1 isoform 1 ( Figure 1A ) and are identical to amino acids 1-19 of human MYCT1 isoform 2.

[0074] In one exemplary aspect, the full-length MYCT1 polypeptide human isoform 1 comprises the amino acid sequence of SEQ ID NO: 145:

[0075] MRTQVYEGLCKNYFSLAVLQRDRIKLLFFDILVFLSVFLLFLLFLVDIMANNTT

[0076] SLGSPWPENFWEDLIMSFTVSMAIGLVLGGFIWAVFICLSRRRRASAPISQWSSSRRSRSSYTHGLNRTGFYRHSGCERRSNLSLASLTFQRQASLEQANSFPRKSSFRASTFHPFLQCPPLPVETESQLVTLPSSNISPTISTSHSLSRPDYWSSNSLRVGLSTPPPPAYESIIKAFPDS.

[0077] In some aspects, the MYCT1 polypeptide human isoform 2 comprises the amino acid sequence of SEQ ID NO: 148: MANNTTSLGSPWPENFWEDLIMSFTVSMAIGLVLGGFIWAVFICLSRRRRASAPISQWSSSRRSRSSYTHGLNRTGFYRHSGCERRSNLSLASLTFQRQASLEQANSFPRKSSFRASTFHPFLQCPPLPVETESQLVTLPSSNISPTISTSHSLSRPDYWSSNSLRVGLSTPPPPAYESIIKAFPDS.

[0078] In some aspects, the human MYCT1 polynucleotide comprises the nucleic acid sequence of SEQ ID NO:149: ATGCGAACACAAGTATATGAGGGGTTGTGTAAAAATTATTTTTCTCTTGCTGTACTACAAAGAGATAGAATCAAACTGCTTTTTTTCGACATACTGGTTTTTCTTTCTGTTTTTCTTCTCTTTCTTCTATTTCTTGTGGATATTATGGCTAATAACACAACAAGTTTAGGGAGTCCATGGCCAGAAAACTTTTGGGAGGACCTTATCATGTCCTTCACTGTATCCATGGCAATCGGGCTGGTACTTGGAGGATTTATTTGGGCTGTGTTCATTTGTCTGTCTCGAAGAAGAAGAGCCAGTGCTCCCATCTCACAGTGGAGTTCAAGCAGGAGATCTAGGTCTTCTTACACCCACGGCCTCAACAGAACTGGATTTTACCGCCACAGTGGCTGTGAACGTCGAAGCAACCTCAGCCTGGCCAGTCTCACCTTCCAGCGACAAGCTTCCCTGGAACAAGCAAATTCCTTTCCAAGAAAATCAAGTTTCAGAGCTTCTACTTTCCATCCCTTTCTGCAATGTCCACCACTTCCTGTGGAAACTGAGAGTCAGCTGGTGACTCTCCCTTCTTCCAATATCTCTCCCACCATCAGCACTTCCCACAGTCTGAGCCGTCCTGACTACTGGTCCAGTAACAGTCTTCGAGTGGGCCTTTCAACACCGCCCCCACCTGCCTATGAGTCCATCATCAAGGCATTCCCAGATTCCTGA。

[0079] In some aspects, the mouse MYCT1 polypeptide comprises the sequence of SEQ ID NO:150: MANNTTSLGSPWPENFWEDLIMSFTVSVAIGLAIGGFLWALFVFLSRRRRASAPISQWSPTRRPRSSYNHGLNRTGFYRHSGYERRSNLSLASLTFQRQASMELVNSFPRKSSFRASTFHPFLQCPPLPVETESQLMTLSASTTPSTLSTAHSPSRPDFRWSSNSLRMGLSTPPPPAYESIIKAFPDS。

[0080] In one exemplary aspect, the full-length MYCT1 mRNA transcript of 3030 base pairs in length includes the NCBI reference sequence: NM_025107.3. Additional reference MYCT1 mRNAs include, but are not limited to, NM_001371624.1nM_001371625.1nM_001371626.1nM_025107.3. In one exemplary aspect, regulatory elements that modulate MYCT1 expression include those described herein and those disclosed in Gene Card ID: GC06P152697.

[0081] The "percent identity" of two amino acid sequences can be determined using the algorithm of Karlin and Altschul Proc. Natl. Acad. Sci. USA 87:2264-68, 1990, as modified in Karlin and Altschul Proc. Natl. Acad. Sci. USA 90:5873-77, 1993. This algorithm is incorporated into the NBLAST and XBLAST programs (version 2.0) of Altschul et al., J. Mol. Biol. 215:403-10, 1990. BLAST protein searches can be performed with the XBLAST program with score = 50 and word length = 3 to obtain amino acid sequences homologous to the protein molecule of interest. When gaps exist between the two sequences, Gapped BLAST can be utilized as described in Altschul et al., Nucleic Acids Res. 25(17):3389-3402, 1997. When utilizing BLAST and Gapped BLAST programs, the default parameters of the respective programs (eg, XBLAST and NBLAST) can be used.

[0082] As used herein, "nucleic acid", "nucleic acid molecule" and "polynucleotide" are used interchangeably herein. The term "nucleic acid encoding..." or "nucleic acid molecule encoding..." is understood to refer to a sequence of nucleotides encoding a polypeptide.

[0083] The polynucleotides described herein may comprise one or more nucleic acids each encoding a polypeptide, all of which may be operably linked to (i.e., have a functional relationship with) one or more regulatory sequences, such as promoters. Such polynucleotides may alternatively be referred to herein as "nucleic acid constructs" or "constructs."

[0084] As used herein, "polypeptide" and "protein" are used interchangeably to refer to a polymer of amino acid residues.

[0085] In further preferred aspects, each amino acid sequence described herein has the following identity or similarity to a given nucleotide or amino acid sequence, respectively: at least 60%, at least 61%, at least 62%, at least 63%, at least 64%, at least 65%, at least 66%, at least 67%, at least 68%, at least 69%, at least 70%, at least 71%, at least 72%, at least 73%, at least 74%, at least 75%, at least 76%, at least 77%, at least 78%, at least 79%, at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity or similarity to a given nucleotide or amino acid sequence, respectively. The terms "homology", "sequence identity" and the like are used interchangeably herein. Sequence identity is described herein as the relationship between two or more amino acid (polypeptide or protein) sequences or two or more nucleic acid (polynucleotide) sequences, as determined by comparing the sequences. In a preferred aspect, sequence identity is calculated based on the full length (in terms of amino acids or nucleotides) of two given SEQ ID NOs or based on a portion thereof. A portion of a full-length sequence may be referred to as a fragment, and preferably means at least 50%, 60%, 70%, 80%, 90% or 100% of the length (in terms of amino acids or nucleotides) of a reference sequence. "Identity" also refers to the degree of sequence relatedness between two amino acid sequences or between two nucleic acid sequences, as the case may be, as determined by the match between such sequence strings. For example, by comparing two sequences using a computer program commonly used for this purpose, such as a global or local alignment algorithm, the degree of sequence identity between the two sequences can be determined. Non-limiting examples include BLASTp, BLASTn, Clustal W, MAFFT, Clustal Omega, AlignMe, Praline, GAP, BESTFIT or another suitable method or algorithm. The Needleman and Wunsch global alignment algorithm can be used to align the two sequences over their entire length or a portion thereof (a portion can mean at least 50%, 60%, 70%, 80%, 90% of the length of the sequence), maximizing the number of matches and minimizing the number of gaps.Default settings can be used, and preferred programs are Needle for pairwise alignments (in one aspect, using EMBOSS Needle 6.6.0.0, start gap open penalty: 10, gap extent penalty: 0.5, end gap penalty: False, end gap open penalty: 10, end gap extension penalty: 0.5) and MAFFT for multiple sequence alignment (in one aspect, using MAFFT v7 defaults are: BLOSUM62 [bl62], start gap: 1.53, gap extension: 0.123, order: aligned, tree rebuilding number: 2, guide tree output: on [true], Max iterate: 2, perform FFTS: no).

[0086] For example, by comparing the amino acid sequence of one polypeptide and its conservative amino acid substitutions with the sequence of a second polypeptide, the "similarity" between two amino acid sequences is determined. Similar algorithms for determining sequence identity can be used to determine sequence similarity. Optionally, when determining the degree of amino acid similarity, the skilled person may also consider so-called conservative amino acid substitutions. As used herein, "conservative" amino acid substitutions refer to the interchangeability of residues with similar side chains. Examples of types of amino acid residues used for conservative substitutions are given in the following table:

[0087] Acidic residue Asp(D) and Glu(E) Basic residues Lys(K), Arg(R) and His(H) Hydrophilic uncharged residues Ser(S), Thr(T), Asn(N) and Gln(Q) Aliphatic uncharged residue Gly(G), Ala(A), Val(V), Leu(L) and Ile(I) Nonpolar uncharged residues Cys(C), Met(M) and Pro(P) Aromatic residues Phe(F), Tyr(Y) and Trp(W)

[0088] Alternative conservative amino acid residue substitution categories:

[0089] 1 A S T 2 D E 3 N Q 4 R K 5 I L M 6 F Y W

[0090] Physical and functional classification of candidate amino acid residues:

[0091] Residues containing alcohol groups S and T Aliphatic residues I, L, V, and M Cycloalkenyl-related residues F, H, W, and Y Hydrophobic residues A, C, F, G, H, I, L, M, R, T, V, W and Y Negatively charged residues D and E Polar residues C, D, E, H, K, N, Q, R, S, and T Positively charged residues H, K and R Small residues A, C, D, G, N, P, S, T and V Very small residue A, G, and S Residues involved in turn formation A, C, D, E, G, H, K, N, Q, R, S, P and T Flexible residues Q, T, K, S, G, P, D, E, and R

[0092] For example, a group of amino acids with aliphatic side chains are glycine, alanine, valine, leucine and isoleucine; a group of amino acids with aliphatic-hydroxy side chains are serine and threonine; a group of amino acids with amide-containing side chains are asparagine and glutamine; a group of amino acids with aromatic side chains are phenylalanine, tyrosine and tryptophan; a group of amino acids with basic side chains are lysine, arginine and histidine; and a group of amino acids with sulfur-containing side chains are cysteine ​​and methionine. Preferred conservative amino acid substitution groups are: valine-leucine-isoleucine, phenylalanine-tyrosine, lysine-arginine, alanine-valine and asparagine-glutamine. Substitution variants of the amino acid sequences disclosed herein are those in which at least one residue in the disclosed sequence is removed and a different residue is inserted in its place. Preferably, the amino acid changes are conservative. Preferred conservative substitutions for each naturally occurring amino acid are as follows: Ala to Ser; Arg to Lys; Asn to Gln or His; Asp to Glu; Cys to Ser or Ala; Gln to Asn; Glu to Asp; Gly to Pro; His to Asn or Gln; Ile to Leu or Val; Leu to Ile or Val; Lys to Arg; Gln or Glu; Met to Leu or Ile; Phe to Met, Leu or Tyr; Ser to Thr; Thr to Ser; Trp to Tyr; Tyr to Trp or Phe; and Val to Ile or Leu.

[0093] As used herein, "fragment" means a portion of a polypeptide that preferably contains at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95% or more of the full length of a reference polypeptide. In some aspects, a fragment has the same or similar biological activity as a reference polypeptide.

[0094] As used herein, the terms "treat," "treating," or "treatment" refer to both therapeutic treatment and prophylactic or preventative measures, wherein the purpose is to prevent or slow down (mitigate) an undesirable physiological change or disease / disorder. Beneficial or desired clinical results include, but are not limited to, relief of symptoms, reduction in the extent of the disease, stabilization of the disease state (i.e., no worsening), delay or slowing of disease progression, improvement or alleviation of the disease state, and relief (whether partial or total), whether detectable or undetectable. "Treatment" may also mean prolonging survival compared to the expected survival if not receiving treatment. Those in need of treatment include those already suffering from a disease, condition, or disorder, as well as those with a tendency to suffer from a disease, condition, or disorder, or those in which a disease, condition, or disorder is to be prevented.

[0095] As used herein, "cancer", "tumor" or "malignant tumor" may refer to one or more tumors or cancers. Tumors may be malignant or benign, cancers may be primary or metastatic; tumors or cancers may be early or advanced. Non-limiting examples of tumors or cancers may include acute lymphoblastic leukemia, acute myeloid leukemia, adrenocortical carcinoma, AIDS-related cancers, AIDS-related lymphomas, anal cancer, appendix cancer, astrocytoma (cerebellum or cerebrum in children), basal cell carcinoma, bile duct cancer, bladder cancer, bone cancer, brain stem glioma, brain tumors (cerebellar astrocytoma, cerebral astrocytoma / malignant glioma, ependymoma, medulloblastoma, supratentorial primitive neuroectodermal tumor, visual pathway and hypothalamic glioma), breast cancer, bronchial adenoma / carcinoid, Burkitt's lymphoma, cerebral ..., cerebral astrocytoma, cerebral astrocytoma / malignant glioma, cerebral astrocytoma / malignant glioma, cerebral astrocytoma / malignant glioma, cerebral astrocytoma / malignant glioma, cerebral astrocytoma / malignant glioma, cerebellar astrocytoma, cerebral astrocytoma / glioblastoma, cervical cancer, childhood cancer, chronic lymphocytic leukemia, chronic myeloid leukemia, chronic myeloproliferative disorders, colon cancer, cutaneous T-cell lymphoma, desmoplastic small round cell tumor, endometrial cancer, ependymoma, esophageal cancer, Ewing sarcoma in the Ewing family of tumors, extracranial germ cell tumor (children), extragonadal germ cell tumor, extrahepatic bile duct cancer, eye cancer (intraocular melanoma, retinal menoblastoma), gallbladder cancer, gastric (stomach) cancer, gastrointestinal carcinoid tumors, gastrointestinal stromal tumors, germ cell tumors (extracranial, extragonadal, ovarian in children), gestational trophoblastic tumors, gliomas (adults, brainstem in children, cerebral astrocytoma in children, visual pathway and hypothalamus in children), gastric carcinoid, hairy cell leukemia, head and neck cancer, hepatocellular (liver) cancer, Hodgkin lymphoma, hypopharyngeal cancer, hypothalamic and visual pathway gliomas (children), intraocular melanoma, islet cell carcinoma, Kaposi's sarcoma, kidney cancer (renal cell carcinoma), laryngeal cancer, leukemia (acute lymphoblastic , acute myeloid, chronic lymphocytic, chronic myeloid, hairy cell), lip and oral cavity cancer, liver cancer (primary), lung cancer (non-small cell, small cell), lymphoma (AIDS-related, Burkitt's, cutaneous T-cell, Hodgkin, non-Hodgkin, primary central nervous system), macroglobulinemia (Waldenstrom's disease), malignant fibrous histiocytoma of bone / osteosarcoma, medulloblastoma (children), melanoma, intraocular melanoma, Merkel cell carcinoma, mesothelioma (malignant in adults, children), metastatic squamous neck cancer with occult primaryoccultprimary), oral cancer, multiple endocrine neoplasia syndrome (children), multiple myeloma / plasmacytoma, mycosis fungoides, myelodysplastic syndrome, myelodysplastic / myeloproliferative disorders, myeloid leukemia (chronic), myeloid leukemia (acute in adults, acute in children), multiple myeloma, myeloproliferative disorders (chronic), nasal and paranasal sinus cancer, nasopharyngeal cancer, neuroblastoma, non-Hodgkin lymphoma, non-small cell lung cancer, oral cancer, oropharyngeal cancer, Osteosarcoma / malignant fibrous histiocytoma of bone, ovarian cancer, ovarian epithelial cancer (surface epithelial-stromal tumor), ovarian germ cell tumor, ovarian low-grade malignant potential tumor, pancreatic cancer, pancreatic cancer (islet cell), paranasal sinus and nasal cavity cancer, parathyroid cancer, penile cancer, pharyngeal cancer, pheochromocytoma, pineal astrocytoma, pineal germ cell tumor, pineoblastoma and supratentorial primitive neuroectodermal tumor (children), pituitary adenoma, plasma cell tumor, pleuropulmonary blastoma, primary central nervous system Lymphoma, prostate cancer, colorectal cancer, renal cell carcinoma (kidney cancer), transitional cell carcinoma of the renal pelvis and ureter, retinoblastoma, rhabdomyosarcoma (children), salivary gland cancer, sarcomas (Ewing family tumors, Kaposi, soft tissue, uterine), Sezary syndrome, skin cancer (non-melanoma, melanoma), skin cancer (Merck cell), small cell lung cancer, small intestine cancer, soft tissue sarcoma, squamous cell carcinoma, squamous neck cancer with occult primary (metastatic), gastric cancer, supratentorial primitive neuroectodermal tumor (children) ), T-cell lymphoma (skin), testicular cancer, laryngeal cancer, thymoma (children), thymoma and thymic carcinoma, thyroid cancer, thyroid cancer (children), transitional cell carcinoma of the renal pelvis and ureter, trophoblastic tumor (pregnancy), unknown primary site (adults, children), transitional cell carcinoma of the ureter and renal pelvis, urethral cancer, uterine cancer (endometrial), uterine sarcoma, vaginal cancer, visual pathway and hypothalamic glioma (children), vulvar cancer, Waldenstrom's macroglobulinemia and Wilms' tumor (children).

[0096] As used herein, the treatment of cancer can include the inhibition of the increase of cancer progression and / or metastasis, the inhibition of tumor volume growth, the reduction of tumor volume and / or growth, the reduction of tumor growth rate, the eradication or killing of tumor and / or cancer cells, or any combination thereof. In some aspects, treatment can also prolong the survival period of the subject, improve the prognosis of the subject, and / or improve the quality of life of the subject.

[0097] As used herein, "resistant" or "insensitive" cancer, tumor cells, and tumors refer to cancers that have developed resistance to cancer therapy (e.g., immune checkpoint inhibitors) and may result in increased tumor volume, cancer metastasis, cancer recurrence, or decreased life expectancy.

[0098] The subject can be a rodent, a human, a livestock animal, a companion animal, or a zoo animal. In one aspect, the subject can be a rodent, such as a mouse, a rat, a guinea pig, etc. In another aspect, the subject can be a livestock animal. Non-limiting examples of suitable livestock animals can include pigs, cattle, horses, goats, sheep, llamas, and alpacas. In yet another aspect, the subject can be a companion animal. Non-limiting examples of companion animals can include pets, such as dogs, cats, rabbits, and birds. In yet another aspect, the subject can be a zoo animal. As used herein, "zoo animals" refers to animals that can be found in zoos. Such animals can include non-human primates, large cats, wolves, and bears. In preferred aspects, the subject is a human.

[0099] As used herein, "control samples" or "control cells" can be obtained from healthy subjects and / or from subjects with cancer (obtained before starting treatment (baseline)). Control subjects are healthy subjects or subjects who have not received treatment. In some aspects, the parameters measured during treatment can be the average or population average of several control subjects. In some aspects, the control sample can include non-cancerous cells. In some aspects, non-cancerous cells can be from the same tissue type as cancer cells. For example, if cancer cells are from breast cancer, non-cancerous cells can be from healthy breast tissue. In some aspects, the control can include the average level of analytes in samples from subjects before the onset of cancer. In some aspects, the control sample can be a sample from a subject before diagnosis or treatment. In some aspects, analytes can be measured in one or more people other than subjects with cancer. In some aspects, the control is one or more people with characteristics similar to those of subjects with cancer. In some aspects, the control can be the average value of a combination of disclosed analyte levels from different healthy sources (e.g., more than one healthy control subject and / or more than one subject before starting treatment (baseline)). In some aspects, the control sample can be a combined sample.

[0100] As used herein, biological sample can be any biological tissue, fluid or cell from a subject. Sample can be solid or fluid. Sample can be a heterogeneous cell colony. Non-limiting examples of suitable biological samples include sputum, serum, blood, blood cells (e.g., leukocytes), biopsy, urine, peritoneal fluid, pleural fluid or cells derived therefrom. Biopsy can be fine needle aspiration biopsy, core needle biopsy, vacuum-assisted biopsy, open surgical biopsy, scrape biopsy, punch biopsy, incisional biopsy, scrape biopsy or deep excavation biopsy. Biological sample can also include tissue sections, such as frozen sections or formalin-fixed sections for histological purposes. Sample can be tumor tissue, tissue around tumor or non-tumor tissue. Methods for obtaining biological samples from subjects are well known in the art.

[0101] As used herein, the term "vector" is understood to be a nucleic acid, which means that it includes a nucleotide sequence that can be introduced into a host cell to recombinantly insert into the genome of the host cell or replicate spontaneously as an episome. Vectors can include linear nucleic acids, plasmids, phagemids, cosmids, RNA vectors, viral vectors and the like. Examples of viral vectors can include retroviruses, adenoviruses, and adeno-associated viruses, but are not limited thereto.

[0102] As used herein, "immune checkpoint inhibitors" or "ICIs" are drugs that block immune checkpoints. These checkpoints are a normal part of the immune system and prevent the immune response from being too strong. By blocking them, these drugs allow immune cells to respond more strongly to, for example, cancer. Immune checkpoint inhibitors work by preventing cancer cells from shutting down T cells (white blood cells that detect infection and abnormalities). Non-limiting examples of immune checkpoint inhibitors include inhibitors of PD-1, PD-L1, TIM-3, LAG-3, CTLA-4, and CSF-1R and any combination thereof. Immune checkpoint receptors can be on tumor cells or immune cells, such as T cells, monocytes, microglia, and macrophages, but are not limited thereto. The agent that acts as an immune checkpoint blockade can be a small chemical entity or polymer, an antibody, an antibody fragment, a single-chain antibody, or other antibody construct, including but not limited to bispecific antibodies and double antibodies. Immune checkpoint inhibitors that can be used according to the present disclosure include any immune checkpoint inhibitors that disrupt the inhibitory interaction between cytotoxic T cells and tumor cells. These include, but are not limited to, anti-PD-1 antibodies, anti-PD-L1 antibodies, anti-CTLA4 antibodies, anti-LAG-3 antibodies, anti-TIM-3 antibodies. The inhibitor need not be an antibody, but can be a small molecule or other polymer. If the inhibitor is an antibody, it can be a polyclonal, monoclonal, fragment, single chain or other antibody variant construct. Inhibitors can target any immune checkpoint known in the art, including but not limited to CTLA-4, PDL1, PDL2, PD1, B7-H3, B7-H4, BTLA, HVEM, TIM3, GAL9, LAG3, CSF-1R, VISTA, KIR, 2B4, CD160, CGEN-15049, CHK1, CHK2, A2aR, CD28, CD 86, CD69, CD48, CD113, CEACAM-1, galectin-1, TIGIT, GPR56, CD48, GARP, PD1H, LAIR1, TIM1, TIM4 and B-7 family ligands. Inhibitors for a single target immune checkpoint or a combination of different inhibitors for different immune checkpoints can be used. Illustrative examples of immune checkpoint inhibitors include CTLA-4 blocking antibodies (ipilimumab (Yervoy), tremelimumab (Imjuno)), PD-1 inhibitors (pembrolizumab (Keytruda), nivolumab (Opdivo), cemiplimab (Libtayo), CT-011 ( Pidilimonoclonal antibody (Pidilizumab), AMP224), PD-L1 inhibitors (Atezolizumab (tecentriq), Avelumab (Bavencio), Durvalumab (Imfinzi), BMS-936559), Lag3 inhibitor (Relatlimab), combination of Lag3 and PD1 inhibitor (PD-1 inhibitor Nivolumab (Opdupa lag), OX40 inhibitors (MEDI6469), CD160 inhibitors (BY55). Non-limiting examples of inhibitors of CSF-1R include PLX3397, PLX486, RG7155, AMG820, ARRY-382, FPA008, IMC-CS4, JNJ-40346527 and MCS110. The terms "ICI treatment", "ICI therapy", "ICI compound" and the like refer to one or more ICIs (or their uses) disclosed herein or known to those skilled in the art.

[0103] As used herein, "angiogenesis" is the growth of blood vessels from an existing vascular system. Angiogenesis can be further defined herein as involving: (i) activation of endothelial cells; (ii) increased vascular permeability; (iii) subsequent dissolution of the basement membrane and extravasation of plasma components, resulting in the formation of a temporary fibrin gel extracellular matrix; (iv) proliferation and migration of endothelial cells; (v) reorganization of migrated endothelial cells to form functional capillaries; (vi) capillary loop formation; (vi) deposition of basement membrane and recruitment of perivascular cells to newly formed blood vessels, or any combination thereof. Normal angiogenesis is active during tissue growth from embryonic development to maturity, and then enters a relatively quiescent period in adulthood. Normal angiogenesis is also activated during wound healing and at certain stages of the female reproductive cycle. Inappropriate or pathological angiogenesis is associated with several disease states, including various retinal diseases, ischemic diseases, atherosclerosis, chronic inflammatory conditions and cancer. Pathological angiogenesis can be the formation or regeneration of blood vessels that are not in a physiologically normal state. Such vessels are immature and diapedetic, and may become vessels that may not be perfused with blood. In addition, pathological angiogenesis may be angiogenesis or neovascularization that occurs in a manner that is different from the physiological normal state. For example, this term may refer to the formation of new blood vessels by receiving stimuli such as inflammation.

[0104] As used herein, "angiogenesis-related diseases or disorders" refer to any disease or disorder that generates the "pathological angiogenesis or neovascularization" described above or causes a functional disorder in an individual due to "pathological angiogenesis or neovascularization". Non-limiting examples of such diseases or disorders include age-related macular degeneration, ischemic retinopathy, intraocular neovascularization, corneal neovascularization, retinal neovascularization, choroidal neovascularization, diabetic macular edema, diabetic retinal ischemia, diabetic retinal edema, diabetic retinopathy, cancer, rheumatoid arthritis, endometriosis, and alopecia. In some aspects, the disease or disorder is cancer.

[0105] As used herein, "VEGF inhibitor" describes an inhibitor of VEGF synthesis or activity, as the case may be, at the protein level, to suppress or downregulate the production of the protein or once produced, to suppress or downregulate at least one biological activity of the protein. In some aspects, the VEGF inhibitor can be selected from: anti-VEGF immunoglobulin, anti-VEGF antibody, anti-VEGF monoclonal antibody and humanized anti-VEGF monoclonal antibody.

[0106] As used herein, "formulation" refers to preparing a drug in a form suitable for administration to a subject (eg, a human). Therefore, a "formulation" may include a pharmaceutically acceptable excipient, including a diluent or carrier.

[0107] As used herein, "pharmaceutically acceptable" describes a substance or component that does not cause unacceptable loss of pharmacological activity or unacceptable adverse side effects. Examples of pharmaceutically acceptable ingredients can be those with monographs in the United States Pharmacopeia (USP29) and National Formulary (NF 24), United States Pharmacopeial Convention, Inc., Rockville, Maryland, 2005 ("USP / NF") or more recent versions, and components listed in the FDA's continuously updated Inactive Ingredient Search online database. Other useful components not described in USP / NF, etc., can also be used.

[0108] As used herein, "pharmaceutically acceptable excipients" as used herein may include any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic agents or absorption delaying agents. The use of such media and agents for pharmaceutically active substances is well known in the art (see generally Remington's Pharmaceutical Sciences (AR Gennaro ed.), 21st edition, ISBN: 0781746736 (2005)). Except for the case where any conventional media or agents are incompatible with the active ingredient, the use of the media or agents in the therapeutic composition is considered. Supplementary active ingredients may also be incorporated into the composition.

[0109] As used herein, a "stable" formulation or composition can refer to a composition that has sufficient stability to permit storage at a convenient temperature, such as from about 0°C to about 60°C, for a commercially reasonable period of time, for example, at least about one day, at least about one week, at least about one month, at least about three months, at least about six months, at least about one year, or at least about two years.

[0110] As used herein, "expression" or "expression level" or "level of expression" refers to the amount of a particular analyte (e.g., MYCT1) present in a sample. The amount can be the concentration, number, ratio, proportion, or percentage of the analyte compared to a control sample or determined using a standard curve. The amount can be an absolute amount or a relative amount.

[0111] As used herein, "combination therapy" refers to the administration of two or more therapeutic agents to treat the therapeutic conditions or disorders described in the present disclosure. Such administration encompasses the co-administration of these therapeutic agents in a substantially simultaneous manner, such as in a single capsule with a fixed ratio of active ingredients or in a plurality of separate capsules of each active ingredient. In addition, such administration also encompasses the use of various types of therapeutic agents in a sequential manner. In either case, the treatment regimen will provide a beneficial effect of the drug combination in treating the conditions or disorders described herein.

[0112] As used herein, "therapeutically effective" or "effective" is intended to qualify the amount of active ingredient for treating a disease or condition or achieving a clinical endpoint. An effective amount can be an amount that achieves the intended purpose. For example, an effective amount of an antibody can be an amount that reduces tumor volume (e.g., shrinks or eliminates a tumor or reduces the number of circulating cancer cells). The determination of an effective amount is well within the capabilities of those skilled in the art, especially in light of the disclosure provided herein.

[0113] II. Anti-MYCT1 Antibodies

[0114] The present disclosure provides antibodies that specifically bind to MYCT1 protein. In some aspects, the antibodies of the present disclosure bind to the epitope of MYCT1 protein. In some aspects, the antibodies described herein can be separated antibodies that bind to the epitope of MYCT1 protein. In some aspects, the separated antibodies herein can bind to the epitope of MYCT1 protein, wherein the MYCT1 protein is a human MYCT1 protein. In some aspects, the separated antibodies herein can bind to the epitope of MYCT1 protein, wherein the MYCT1 protein is a mouse MYCT1 protein.

[0115] In certain aspects, antibodies are provided herein that bind to the extracellular domain of MYCT1. In some aspects, antibodies are provided herein that specifically bind to an epitope within the first 150 amino acids of a MYCT1 protein. In some aspects, the antibody binds to an epitope within the first 100 amino acids of MYCT1. In some aspects, the antibody binds to an epitope within the first 70 amino acids of MYCT1. In some aspects, the antibody binds to an epitope within the first 67 amino acids of MYCT1, which comprises an amino acid sequence of SEQ ID NO: 145. In some aspects, the antibody binds to an epitope within the first 30 amino acids of MYCT1, which comprises an amino acid sequence of SEQ ID NO: 148 or SEQ ID NO: 150. In some aspects, the antibody binds to an epitope within the first 20 amino acids of MYCT1, which comprises an amino acid sequence of SEQ ID NO: 148 or SEQ ID NO: 150. In some aspects, the epitope comprises an amino acid sequence of SEQ ID NO: 146.

[0116] In certain aspects, the antibodies described herein may have suitable binding affinity for a target antigen, such as a MYCT1 protein or an epitope thereof. As used herein, "binding affinity" refers to an apparent association constant or K A . K A is the dissociation constant (K D In some aspects, the antibodies described herein may have a binding affinity (K) of at least about 100 nM, at least about 10 nM, at least about 1 nM, at least about 0.1 nM or less for the MYCT1 protein or an epitope thereof. D In some aspects, the antibodies described herein may have a binding affinity (K) of about 0.1 pM to about 10 μM, preferably about 0.1 pM to about 1 μM, more preferably about 0.1 pM to about 100 nM. D In some aspects, the antibodies described herein may have the following binding affinity (K) for MYCT1 protein or its epitope: D): about 100nM to about 0.1nM (e.g., about 100nM, about 75nM, about 50nM, about 25nM, about 10nM, about 5nM, about 1nM, about 0.75nM, about 0.5nM, about 0.25nM, about 0.1nM). In some aspects, the antibodies described herein may have the following binding affinity (K) for MYCT1 protein or its epitope: D ): about 50nM to about 40nM (e.g., about 50nM, about 49nM, about 48nM, about 47nM, about 46nM, about 45nM, about 44nM, about 43nM, about 42nM, about 41nM, about 40nM). In some aspects, the antibodies described herein may have the following binding affinity (K) for MYCT1 protein or its epitope: D ): about 50nM to about 40nM (e.g., about 50nM, about 49nM, about 48nM, about 47nM, about 46nM, about 45nM, about 44nM, about 43nM, about 42nM, about 41nM, about 40nM). In some aspects, binding affinity (or binding specificity) can be determined by various methods, including equilibrium dialysis, equilibrium binding, gel filtration, ELISA, surface plasmon resonance and / or spectroscopy (e.g., using fluorescence).

[0117] In some aspects, antibodies described herein can have neutralizing activity for MYCT1 protein. In some aspects, antibodies described herein can have neutralizing activity for MYCT1 protein, wherein IC50 is less than about 150ng / mL. In some aspects, antibodies described herein can have neutralizing activity for MYCT1 protein, wherein IC50 is in the range of about 0.1ng / ml to about 150ng / mL (e.g., about 0.1ng / ml, about 0.25ng / ml, about 0.5ng / ml, about 1ng / ml, about 2.5ng / ml, about 5ng / ml, about 7.5ng / ml, about 10ng / ml, about 25ng / ml, about 50ng / ml, about 75ng / ml, about 100ng / ml, about 125ng / ml, about 150ng / ml).

[0118] In some aspects, compared with the MYCT1 expression and / or activity of the antibody not present herein, the antibody described herein can reduce the expression and / or activity of MYCT1. In certain aspects, compared with the MYCT1 expression and / or activity of the antibody not present herein, the antibody described herein can reduce the expression and / or activity of MYCT1 by about 5% to about 99% (e.g., about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 99%).

[0119] In some aspects, MYCT1 expression can be measured using methods known in the art. By way of non-limiting example, the expression level of MYCT1 can be measured using RNA-seq, nanopore sequencing, Nanostring, multiple RT-PCR, single RT-PCR, NASBA, fluorescence measurement or spectrophotometry. MYCT 1 interacts with tight junction protein Zona Occludens 1, and regulates the actin cytoskeleton dynamics mediated by Rho GTPase, thereby promoting endothelial cell movement. Therefore, in some aspects, actin cytoskeleton dynamics can be measured as an indication of MYCT1 activity. Actin cytoskeleton dynamics and MYCT1 activity can be measured using standard methods in the art as described below in the examples. On the other hand, tubular structure formation assays in Matrigel, Boyden chamber tumor chemotaxis assays, wound closure assays and cell morphology assays can be measured as an indication of MYCT1 activity.

[0120] In some aspects, antibody described herein can have the same heavy chain CDR and / or light chain CDR as any exemplary antibody described herein (e.g., antibody clone 2B4, 9F8, 8D2).Two antibodies with the same heavy chain CDR and / or light chain CDR mean that when measured by the same method (e.g., Kabat method, Chothia method, AbM method, contact method or IMGT method), their CDR is identical. In some aspects, antibody described herein can have the same heavy chain and / or light chain as any exemplary antibody described herein. In certain embodiments, antibody disclosed herein can share one or more amino acid sequences provided in Table 7.

[0121] In some aspects, individually or collectively compared to the heavy chain of the exemplary antibodies described herein, the antibodies herein may include a heavy chain with at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) sequence identity. In some aspects, individually or collectively compared to the light chain of the exemplary antibodies described herein, the antibodies herein may include a light chain with at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) sequence identity. In some aspects, individually or collectively compared to the heavy chain CDR of the exemplary antibodies described herein, the antibodies herein may include a heavy chain CDR with at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) sequence identity. In some aspects, individually or collectively compared to the light chain CDR of the exemplary antibodies described herein, the antibodies herein may include a light chain CDR with at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) sequence identity.

[0122] In some aspects, the isolated anti-MYCT1 antibody may comprise a heavy chain variable region (VH) comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of any one of SEQ ID NOs: 18, 54, 90, or 126. In some aspects, the anti-MYCT1 antibody comprises a VH comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 18. In some aspects, the anti-MYCT1 antibody comprises a VH comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 54. In some aspects, the anti-MYCT1 antibody comprises a VH comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 90. In some aspects, the anti-MYCT1 antibody comprises a VH comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO:126.

[0123] In some aspects, the isolated anti-MYCT1 antibody may comprise a VH comprising the amino acid sequence of any one of SEQ ID NOs: 18, 54, 90, or 126. In some aspects, the anti-MYCT1 antibody comprises a VH comprising the amino acid sequence of SEQ ID NO: 18. In some aspects, the anti-MYCT1 antibody comprises a VH comprising the amino acid sequence of SEQ ID NO: 54. In some aspects, the anti-MYCT1 antibody comprises a VH comprising the amino acid sequence of SEQ ID NO: 90. In some aspects, the anti-MYCT1 antibody comprises a VH comprising the amino acid sequence of SEQ ID NO: 126.

[0124] In some aspects, the isolated anti-MYCT1 antibody may comprise a light chain variable region (VL) comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of any one of SEQ ID NOs: 36, 72, 108, or 144. In some aspects, the isolated anti-MYCT1 antibody may comprise a VL comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 36. In some aspects, the isolated anti-MYCT1 antibody may comprise a VL comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 72. In some aspects, the isolated anti-MYCT1 antibody may comprise a VL comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 108. In some aspects, the isolated anti-MYCT1 antibody may comprise a VL comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 144.

[0125] In some aspects, the isolated anti-MYCT1 antibody may comprise a VL comprising an amino acid sequence of any one of SEQ ID NOs: 36, 72, 108, or 144. In some aspects, the isolated anti-MYCT1 antibody may comprise a VL comprising an amino acid sequence of SEQ ID NO: 36. In some aspects, the isolated anti-MYCT1 antibody may comprise a VL comprising an amino acid sequence of SEQ ID NO: 72. In some aspects, the isolated anti-MYCT1 antibody may comprise a VL comprising an amino acid sequence of SEQ ID NO: 108. In some aspects, the isolated anti-MYCT1 antibody may comprise a VL comprising an amino acid sequence of SEQ ID NO: 144.

[0126] In some aspects, the isolated anti-MYCT1 antibody may comprise a VH comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to an amino acid sequence of any one of SEQ ID NOs: 18, 54, 90, or 126; the VL comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to an amino acid sequence of any one of SEQ ID NOs: 36, 72, 108, or 144, or any combination thereof. In some aspects, the isolated anti-MYCT1 antibody may comprise a VH, a VL, a VH, comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 18; the VL, the VL, comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 36. In some aspects, the isolated anti-MYCT1 antibody may comprise a VH, a VL, a VH, comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 54; the VL, the VL, comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 72. In some aspects, the isolated anti-MYCT1 antibody may comprise a VH, wherein the VL, wherein the VH comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 90; wherein the VL, wherein the VL comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 108. In some aspects, the isolated anti-MYCT1 antibody may comprise a VH, wherein the VL, wherein the VH comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 126; wherein the VL, wherein the VL comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 144.

[0127] In some aspects, the isolated anti-MYCT1 antibody may comprise a VH and a VL, the VH comprising the amino acid sequence of any one of SEQ ID NOs: 18, 54, 90 or 126; the VL comprising the amino acid sequence of any one of SEQ ID NOs: 36, 72, 108 or 144, or any combination thereof. In some aspects, the isolated anti-MYCT1 antibody may comprise a VH and a VL, the VH comprising the amino acid sequence of SEQ ID NO: 18; the VL comprising the sequence of SEQ ID NO: 36. In some aspects, the isolated anti-MYC T1 antibody may comprise a VH and a VL, the VH comprising the amino acid sequence of SEQ ID NO: 54; the VL comprising the amino acid sequence of SEQ ID NO: 72. In some aspects, the isolated anti-MYCT1 antibody may comprise a VH and a VL, the VH comprising the sequence of SEQ ID NO: 90; the VL comprising the amino acid sequence of SEQ ID NO: 108. In some aspects, the isolated anti-MYCT1 antibody may comprise a VH comprising the amino acid sequence of SEQ ID NO: 126 and a VL comprising the amino acid sequence of SEQ ID NO: 144.

[0128] In some aspects, the isolated anti-MYCT1 antibody may comprise a heavy chain variable region 1 (H1) comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of any one of SEQ ID NOs: 10, 46, 82, or 118. In some aspects, the isolated anti-MYCT1 antibody may comprise an H1 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 10. In some aspects, the isolated anti-MYCT1 antibody may comprise an H1 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 46. In some aspects, the isolated anti-MYCT1 antibody may comprise an H1 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 82. In some aspects, the isolated anti-MYCT1 antibody may comprise an H1 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 118.

[0129] In some aspects, the isolated anti-MYCT1 antibody may comprise a heavy chain variable region 1 (H1) comprising an amino acid sequence of any one of SEQ ID NOs: 10, 46, 82, or 118. In some aspects, the isolated anti-MYCT1 antibody may comprise an H1 comprising an amino acid sequence of SEQ ID NO: 10. In some aspects, the isolated anti-MYCT1 antibody may comprise an H1 comprising an amino acid sequence of SEQ ID NO: 46. In some aspects, the isolated anti-MYCT1 antibody may comprise an H1 comprising an amino acid sequence of SEQ ID NO: 82. In some aspects, the isolated anti-MYCT1 antibody may comprise an H1 comprising an amino acid sequence of SEQ ID NO: 118.

[0130] In some aspects, the isolated anti-MYCT1 antibody may comprise a light chain variable region 1 (L1) comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of any one of SEQ ID NOs: 28, 64, 100, or 136. In some aspects, the isolated anti-MYCT1 antibody may comprise an L1 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 28. In some aspects, the isolated anti-MYCT1 antibody may comprise an L1 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 64. In some aspects, the isolated anti-MYCT1 antibody may comprise an L1 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 100. In some aspects, the isolated anti-MYCT1 antibody may comprise an L1 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 136.

[0131] In some aspects, the isolated anti-MYCT1 antibody may comprise L1 comprising the amino acid sequence of any one of SEQ ID NO: 28, 64, 100 or 136. In some aspects, the isolated anti-MYCT1 antibody may comprise L1 comprising the amino acid sequence of SEQ ID NO: 28. In some aspects, the isolated anti-MY CT1 antibody may comprise L1 comprising the amino acid sequence of SEQ ID NO: 64. In some aspects, the isolated anti-MYCT1 antibody may comprise L1 comprising the amino acid sequence of SEQ ID NO: 100. In some aspects, the isolated anti-MYCT1 antibody may comprise L1 comprising the amino acid sequence of SEQ ID NO: 136.

[0132] In some aspects, the isolated anti-MYCT1 antibody of the present invention may comprise H1 and L1, wherein H1 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of any one of SEQ ID NOs: 10, 46, 82 or 118; and L1 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of any one of SEQ ID NOs: 28, 64, 100 or 136, or any combination thereof. In some aspects, the isolated anti-MYCT1 antibody herein may comprise H1 and L1, wherein H1 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the sequence of SEQ ID NO: 10; wherein L1 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 28. In some aspects, the isolated anti-MYCT1 antibody herein may comprise H1 and L1, wherein H1 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 46; wherein L1 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 64. In some aspects, the isolated anti-MYCT1 antibody herein may comprise H1 and L1, wherein H1 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 82; wherein L1 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 100. In some aspects, the isolated anti-MYCT1 antibody herein may comprise H1 and L1, wherein H1 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 118; wherein L1 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 136.

[0133] In some aspects, the isolated anti-MYCT1 antibody herein may comprise H1 and L1, wherein H1 comprises the amino acid sequence of any one of SEQ ID NO: 10, 46, 82 or 118; wherein L1 comprises the amino acid sequence of any one of SEQ ID NO: 28, 64, 100 or 136, or any combination thereof. In some aspects, the isolated anti-MYCT1 antibody herein may comprise H1 and L1, wherein H1 comprises the amino acid sequence of SEQ ID NO: 10; wherein L1 comprises the amino acid sequence of SEQ ID NO: 28. In some aspects, the isolated anti-MYCT1 antibody herein may comprise H1 and L1, wherein H1 comprises the amino acid sequence of SEQ ID NO: 46; wherein L1 comprises the amino acid sequence of SEQ ID NO: 64. In some aspects, the isolated anti-MYCT1 antibody herein may comprise H1 and L1, wherein H1 comprises the amino acid sequence of SEQ ID NO: 82; wherein L1 comprises the amino acid sequence of SEQ ID NO: 100. In some aspects, the isolated anti-MYCT1 antibody herein may comprise H1 and L1, wherein H1 comprises the sequence of SEQ ID NO:118; and L1 comprises the amino acid sequence of SEQ ID NO:136.

[0134] In some aspects, the isolated anti-MYCT1 antibody may comprise a heavy chain variable region 2 (H2) comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of any one of SEQ ID NOs: 12, 48, 84, or 120. In some aspects, the isolated anti-MYCT1 antibody may comprise an H2 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 12. In some aspects, the isolated anti-MYCT1 antibody may comprise an H2 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 48. In some aspects, the isolated anti-MYCT1 antibody may comprise an H2 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 84. In some aspects, the isolated anti-MYCT1 antibody may comprise an H2 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 120.

[0135] In some aspects, the isolated anti-MYCT1 antibody may comprise H2 comprising the amino acid sequence of any one of SEQ ID NO: 12, 48, 84 or 120. In some aspects, the isolated anti-MYCT1 antibody may comprise H2 comprising the amino acid sequence of SEQ ID NO: 12. In some aspects, the isolated anti-MYCT1 antibody may comprise H2 comprising the amino acid sequence of SEQ ID NO: 48. In some aspects, the isolated anti-MYCT1 antibody may comprise H2 comprising the amino acid sequence of SEQ ID NO: 84. In some aspects, the isolated anti-MYCT1 antibody may comprise H2 comprising the amino acid sequence of SEQ ID NO: 120.

[0136] In some aspects, the isolated anti-MYCT1 antibody may comprise a light chain variable region 2 (L2) comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of any one of SEQ ID NOs: 30, 66, 102, or 138. In some aspects, the isolated anti-MYCT1 antibody may comprise an L2 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 30. In some aspects, the isolated anti-MYCT1 antibody may comprise an L2 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 66. In some aspects, the isolated anti-MYCT1 antibody may comprise an L2 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 102. In some aspects, the isolated anti-MYCT1 antibody may comprise an L2 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 138.

[0137] In some aspects, the isolated anti-MYCT1 antibody may comprise L2 comprising the amino acid sequence of any one of SEQ ID NO:30, 66, 102 or 138. In some aspects, the isolated anti-MYCT1 antibody may comprise L2 comprising the amino acid sequence of SEQ ID NO:30. In some aspects, the isolated anti-MY CT1 antibody may comprise L2 comprising the amino acid sequence of SEQ ID NO:66. In some aspects, the isolated anti-MYCT1 antibody may comprise L2 comprising the amino acid sequence of SEQ ID NO:102. In some aspects, the isolated anti-MYCT1 antibody may comprise L2 comprising the amino acid sequence of SEQ ID NO:138.

[0138] In some aspects, the isolated anti-MYCT1 antibody of the present invention may comprise H2 and L2, wherein H2 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of any one of SEQ ID NOs: 12, 48, 84 or 120; and L2 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of any one of SEQ ID NOs: 30, 66, 102 or 138, or any combination thereof. In some aspects, the isolated anti-MYCT1 antibody herein may comprise H2 and L2, wherein H2 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 12; wherein L2 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 30. In some aspects, the isolated anti-MYCT1 antibody herein may comprise H2 and L2, wherein H2 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the sequence of SEQ ID NO: 48; wherein L2 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 66. In some aspects, the isolated anti-MYCT1 antibody herein may comprise H2 and L2, wherein H2 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 84; wherein L2 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the sequence of SEQ ID NO: 102. In some aspects, the isolated anti-MYCT1 antibody herein may comprise H2 and L2, wherein H2 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 120; wherein L2 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 138.

[0139] In some aspects, the isolated anti-MYCT1 antibody herein may comprise H2 and L2, wherein H2 comprises the amino acid sequence of any one of SEQ ID NO: 12, 48, 84 or 120; wherein L2 comprises the amino acid sequence of any one of SEQ ID NO: 30, 66, 102 or 138, or any combination thereof. In some aspects, the isolated anti-MYCT1 antibody herein may comprise H2 and L2, wherein H2 comprises the amino acid sequence of SEQ ID NO: 12; wherein L2 comprises the amino acid sequence of SEQ ID NO: 30. In some aspects, the isolated anti-MYCT1 antibody herein may comprise H2 and L2, wherein H2 comprises the sequence of SEQ ID NO: 48; wherein L2 comprises the amino acid sequence of SEQ ID NO: 66. In some aspects, the isolated anti-MYCT1 antibody herein may comprise H2 and L2, wherein H2 comprises the amino acid sequence of SEQ ID NO: 84; wherein L2 comprises the sequence of SEQ ID NO: 102. In some aspects, the isolated anti-MYC T1 antibody herein may comprise H2 and L2, wherein H2 comprises the amino acid sequence of SEQ ID NO:120; and L2 comprises the amino acid sequence of SEQ ID NO:138.

[0140] In some aspects, the isolated anti-MYCT1 antibody may comprise a heavy chain variable region 3 (H3) comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of any one of SEQ ID NOs: 14, 50, 86, or 122. In some aspects, the isolated anti-MYCT1 antibody may comprise an H3 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 14. In some aspects, the isolated anti-MYCT1 antibody may comprise an H3 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 50. In some aspects, the isolated anti-MYCT1 antibody may comprise an H3 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 86. In some aspects, the isolated anti-MYCT1 antibody may comprise an H3 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 122.

[0141] In some aspects, the isolated anti-MYCT1 antibody may comprise H3 comprising the amino acid sequence of any one of SEQ ID NO: 14, 50, 86 or 122. In some aspects, the isolated anti-MYCT1 antibody may comprise H3 comprising the amino acid sequence of SEQ ID NO: 14. In some aspects, the isolated anti-MYCT1 antibody may comprise H3 comprising the amino acid sequence of SEQ ID NO: 50. In some aspects, the isolated anti-MYCT1 antibody may comprise H3 comprising the amino acid sequence of SEQ ID NO: 86. In some aspects, the isolated anti-MYCT1 antibody may comprise H3 comprising the amino acid sequence of SEQ ID NO: 122.

[0142] In some aspects, the isolated anti-MYCT1 antibody may comprise a light chain variable region 3 (L3) comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of any one of SEQ ID NOs: 32, 68, 104, or 140. In some aspects, the isolated anti-MYCT1 antibody may comprise an L3 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 32. In some aspects, the isolated anti-MYCT1 antibody may comprise an L3 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 68. In some aspects, the isolated anti-MYCT1 antibody may comprise an L3 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 104. In some aspects, the isolated anti-MYCT1 antibody may comprise an L3 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 140.

[0143] In some aspects, the isolated anti-MYCT1 antibody may comprise L3 comprising the amino acid sequence of any one of SEQ ID NO: 32, 68, 104 or 140. In some aspects, the isolated anti-MYCT1 antibody may comprise L3 comprising the amino acid sequence of SEQ ID NO: 32. In some aspects, the isolated anti-MY CT1 antibody may comprise L3 comprising the amino acid sequence of SEQ ID NO: 68. In some aspects, the isolated anti-MYCT1 antibody may comprise L3 comprising the amino acid sequence of SEQ ID NO: 104. In some aspects, the isolated anti-MYCT1 antibody may comprise L3 comprising the amino acid sequence of SEQ ID NO: 140.

[0144] In some aspects, the isolated anti-MYCT1 antibody of the present invention may comprise H3 and L3, wherein H3 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of any one of SEQ ID NOs: 14, 50, 86 or 122; and L3 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of any one of SEQ ID NOs: 32, 68, 104 or 140, or any combination thereof. In some aspects, the isolated anti-MYCT1 antibody herein may comprise H3 and L3, wherein H3 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 14; wherein L3 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 32. In some aspects, the isolated anti-MYCT1 antibody herein may comprise H3 and L3, wherein H3 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 50; wherein L3 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the sequence of SEQ ID NO: 68. In some aspects, the isolated anti-MYCT1 antibody herein may comprise H3 and L3, wherein H3 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 86; wherein L3 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 104. In some aspects, the isolated anti-MYCT1 antibody herein may comprise H3 and L3, wherein H3 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 122; wherein L3 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 140.

[0145] In some aspects, the isolated anti-MYCT1 antibody herein may comprise H3 and L3, the H3 comprising the amino acid sequence of any one of SEQ ID NO: 14, 50, 86 or 122; the L3 comprising the amino acid sequence of any one of SEQ ID NO: 32, 68, 104 or 140, or any combination thereof. In some aspects, the isolated anti-MYCT1 antibody herein may comprise H3 and L3, the H3 comprising the amino acid sequence of SEQ ID NO: 14; the L3 comprising the sequence of SEQ ID NO: 32. In some aspects, the isolated anti-MYCT1 antibody herein may comprise H3 and L3, the H3 comprising the amino acid sequence of SEQ ID NO: 50; the L3 comprising the amino acid sequence of SEQ ID NO: 68. In some aspects, the isolated anti-MYCT1 antibody herein may comprise H3 and L3, the H3 comprising the amino acid sequence of SEQ ID NO: 86; the L3 comprising the amino acid sequence of SEQ ID NO: 104. In some aspects, the isolated anti-MYCT1 antibody herein may comprise H3 and L3, wherein H3 comprises the amino acid sequence of SEQ ID NO: 122; and L3 comprises the amino acid sequence of SEQ ID NO: 140.

[0146] In some aspects, the isolated anti-MYCT1 antibody of the present invention may comprise H1, H2 and H3, wherein H1 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of any one of SEQ ID NOs: 10, 46, 82 or 118; the H2 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of any one of SEQ ID NOs: 12, 48, 84 or 120; the H3 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of any one of SEQ ID NOs: 14, 50, 86 or 122, or any combination thereof. In some aspects, the isolated anti-MYCT1 antibody herein may comprise H1, H2 and H3, wherein H1 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 10; the H2 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 12; the H3 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 14, or any combination thereof. In some aspects, the isolated anti-MYCT1 antibody herein may comprise H1, H2 and H3, wherein H1 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO:46; the H2 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO:48; the H3 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO:50, or any combination thereof. In some aspects, the isolated anti-MY CT1 antibody of the present invention may comprise H1, H2 and H3, wherein H1 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 82; the H2 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 84; the H3 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 86, or any combination thereof.In some aspects, the isolated anti-MYCT1 antibody herein may comprise H1, H2 and H3, wherein H1 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 118; the H2 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 120; the H3 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 122, or any combination thereof.

[0147] In some aspects, the isolated anti-MYCT1 antibody herein may comprise H1, H2 and H3, wherein H1 comprises an amino acid sequence of any one of SEQ ID NO: 10, 46, 82 or 118; wherein H2 comprises an amino acid sequence of any one of SEQ ID NO: 12, 48, 84 or 120; wherein H3 comprises an amino acid sequence of any one of SEQ ID NO: 14, 50, 86 or 122, or any combination thereof. In some aspects, the isolated anti-MYCT1 antibody herein may comprise H1, H2 and H3, wherein H1 comprises an amino acid sequence of SEQ ID NO: 10; wherein H2 comprises an amino acid sequence of SEQ ID NO: 12; wherein H3 comprises an amino acid sequence of SEQ ID NO: 14, or any combination thereof. In some aspects, the isolated anti-MYCT1 antibody herein may comprise H1, H2 and H3, wherein H1 comprises the amino acid sequence of SEQ ID NO: 46; the H2 comprises the amino acid sequence of SEQ ID NO: 48; the H3 comprises the amino acid sequence of SEQ ID NO: 50, or any combination thereof. In some aspects, the isolated anti-MYCT1 antibody herein may comprise H1, H2 and H3, wherein H1 comprises the amino acid sequence of SEQ ID NO: 82; the H2 comprises the amino acid sequence of SEQ ID NO: 84; the H3 comprises the amino acid sequence of SEQ ID NO: 86, or any combination thereof. In some aspects, the isolated anti-MYCT1 antibody herein may comprise H1, H2 and H3, wherein H1 comprises the amino acid sequence of SEQ ID NO: 118; the H2 comprises the amino acid sequence of SEQ ID NO: 120; the H3 comprises the amino acid sequence of SEQ ID NO: 122, or any combination thereof.

[0148] In some aspects, the isolated anti-MYCT1 antibody of the present invention may comprise L1, L2 and L3, wherein L1 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of any one of SEQ ID NOs: 28, 64, 100 or 136; the L2 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of any one of SEQ ID NOs: 30, 66, 102 or 138; the L3 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of any one of SEQ ID NOs: 32, 68, 104 or 140, or any combination thereof. In some aspects, the isolated anti-MYCT1 antibody herein may comprise L1, L2 and L3, wherein L1 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 28; the L2 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 30; the L3 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 32, or any combination thereof. In some aspects, the isolated anti-MYCT1 antibody herein may comprise L1, L2 and L3, wherein L1 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO:64; the L2 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO:66; the L3 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO:68, or any combination thereof. In some aspects, the isolated anti-MYCT1 antibody herein may comprise L1, L2 and L3, wherein L1 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 100; the L2 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 102; the L3 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 104, or any combination thereof.In some aspects, the isolated anti-MYCT1 antibody herein may comprise L1, L2 and L3, wherein L1 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 136; the L2 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 138; the L3 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 140, or any combination thereof.

[0149] In some aspects, the isolated anti-MYCT1 antibody herein may comprise L1, L2 and L3, wherein L1 comprises an amino acid sequence of any one of SEQ ID NO: 28, 64, 100 or 136; wherein L2 comprises an amino acid sequence of any one of SEQ ID NO: 30, 66, 102 or 138; wherein L3 comprises an amino acid sequence of any one of SEQ ID NO: 32, 68, 104 or 140, or any combination thereof. In some aspects, the isolated anti-MYCT1 antibody herein may comprise L1, L2 and L3, wherein L1 comprises an amino acid sequence of SEQ ID NO: 28; wherein L2 comprises an amino acid sequence of SEQ ID NO: 30; wherein L3 comprises an amino acid sequence of SEQ ID NO: 32, or any combination thereof. In some aspects, the isolated anti-MYCT1 antibody herein may comprise L1, L2 and L3, wherein L1 comprises the amino acid sequence of SEQ ID NO: 64; wherein L2 comprises the sequence of SEQ ID NO: 66; wherein L3 comprises the amino acid sequence of SEQ ID NO: 68, or any combination thereof. In some aspects, the isolated anti-MYCT1 antibody herein may comprise L1, L2 and L3, wherein L1 comprises the amino acid sequence of SEQ ID NO: 100; wherein L2 comprises the sequence of SEQ ID NO: 102; wherein L3 comprises the amino acid sequence of SEQ ID NO: 104, or any combination thereof. In some aspects, the isolated anti-MYCT1 antibody herein may comprise L1, L2 and L3, wherein L1 comprises the amino acid sequence of SEQ ID NO: 136; wherein L2 comprises the sequence of SEQ ID NO: 138; wherein L3 comprises the amino acid sequence of SEQ ID NO: 140, or any combination thereof.

[0150] In some aspects, the isolated anti-MYCT1 antibody herein may comprise H1, H2, H3, L1, L2 and L3, wherein H1 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to an amino acid sequence of any one of SEQ ID NOs: 10, 46, 82 or 118; the H2 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to an amino acid sequence of any one of SEQ ID NOs: 12, 48, 84 or 120; the H3 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to an amino acid sequence of any one of SEQ ID NOs: 14, 50, 86 or 122; the L1 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to an amino acid sequence of any one of SEQ ID NOs: 14, 50, 86 or 122; NO:28, 64, 100 or 136, any one of at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity; the L2 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity with the amino acid sequence of any one of SEQ ID NO:30, 66, 102 or 138; the L3 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity with the amino acid sequence of any one of SEQ ID NO:32, 68, 104 or 140, or any combination thereof. In some aspects, the isolated anti-MYCT1 antibody herein may comprise H1, H2, H3, L1, L2 and L3, wherein H1 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 10; the H2 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 12; the H3 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of any one of SEQ ID NO: 14; the L1 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 28; the L2 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 29; NO:30 has at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity; said L3 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO:32, or any combination thereof.In some aspects, the isolated anti-MYCT1 antibody herein may comprise H1, H2, H3, L1, L2 and L3, wherein H1 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 46; the H2 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 48; the H3 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of any one of SEQ ID NO: 50; the L1 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the sequence of SEQ ID NO: 64; the L2 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 65; NO:66; the L3 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity with the amino acid sequence of SEQ ID NO:68, or any combination thereof. In some aspects, the isolated anti-MYCT1 antibody herein may comprise H1, H2, H3, L1, L2 and L3, wherein H1 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 82; the H2 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 84; the H3 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of any one of SEQ ID NO: 86; the L1 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 100; the L2 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 101; NO:102; the L3 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO:104, or any combination thereof.In some aspects, the isolated anti-MYCT1 antibody herein may comprise H1, H2, H3, L1, L2 and L3, wherein H1 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 118; the H2 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 120; the H3 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of any one of SEQ ID NO: 122; the L1 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 136; the L2 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 137; NO:138; the L3 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO:140, or any combination thereof.

[0151] In some aspects, the isolated anti-MYCT1 antibody herein may comprise H1, H2, H3, L1, L2 and L3, wherein H1 comprises the amino acid sequence of any one of SEQ ID NO: 10, 46, 82 or 118; the H2 comprises the amino acid sequence of any one of SEQ ID NO: 12, 48, 84 or 120; the H3 comprises the amino acid sequence of any one of SEQ ID NO: 14, 50, 86 or 122; the L1 comprises the amino acid sequence of any one of SEQ ID NO: 28, 64, 100 or 136; the L2 comprises the amino acid sequence of any one of SEQ ID NO: 30, 66, 102 or 138; the L3 comprises the amino acid sequence of any one of SEQ ID NO: 32, 68, 104 or 140, or any combination thereof. In some aspects, the isolated anti-MYCT1 antibody herein may include H1, H2, H3, L1, L2 and L3, wherein H1 includes the amino acid sequence of SEQ ID NO: 10; H2 includes the amino acid sequence of SEQ ID NO: 12; H3 includes the amino acid sequence of any one of SEQ ID NO: 14; L1 includes the amino acid sequence of SEQ ID NO: 28; L2 includes the sequence of SEQ ID NO: 30; L3 includes the sequence of SEQ ID NO: 32, or any combination thereof. In some aspects, the isolated anti-MYCT1 antibody herein may include H1, H2, H3, L1, L2 and L3, wherein H1 includes the amino acid sequence of SEQ ID NO: 46; H2 includes the amino acid sequence of SEQ ID NO: 48; H3 includes the sequence of any one of SEQ ID NO: 50; L1 includes the amino acid sequence of SEQ ID NO: 64; L2 includes the sequence of SEQ ID NO: 66; L3 includes the amino acid sequence of SEQ ID NO: 68, or any combination thereof. In some aspects, the isolated anti-MYCT1 antibody herein may comprise H1, H2, H3, L1, L2 and L3, wherein H1 comprises the amino acid sequence of SEQ ID NO:82; H2 comprises the sequence of SEQ ID NO:84; H3 comprises the amino acid sequence of any one of SEQ ID NO:86; L1 comprises the sequence of SEQ ID NO:100; L2 comprises the amino acid sequence of SEQ ID NO:102; L3 comprises the amino acid sequence of SEQ ID NO:104, or any combination thereof.In some aspects, the isolated anti-MYCT1 antibody herein may comprise H1, H2, H3, L1, L2 and L3, wherein H1 comprises the amino acid sequence of SEQ ID NO: 118; the H2 comprises the amino acid sequence of SEQ ID NO: 120; the H3 comprises the amino acid sequence of any one of SEQ ID NO: 122; the L1 comprises the amino acid sequence of SEQ ID NO: 136; the L2 comprises the amino acid sequence of SEQ ID NO: 138; the L3 comprises the amino acid sequence of SEQ ID NO: 140, or any combination thereof.

[0152] In another exemplary aspect, the anti-MYCT1 antibody is one or more of the antibodies listed in Table 1.

[0153] Table 1: Exemplary anti-MYCT1 antibodies

[0154]

[0155]

[0156]

[0157]

[0158]

[0159]

[0160]

[0161]

[0162]

[0163]

[0164]

[0165]

[0166]

[0167]

[0168]

[0169]

[0170]

[0171]

[0172]

[0173]

[0174] In some aspects, the heavy chain of any antibody disclosed herein may further include a heavy chain constant region (CH) or a portion thereof (e.g., CH1, CH2, CH3, or a combination thereof). In some aspects, the heavy chain constant region used herein may be any suitable source, such as human, mouse, rat, or rabbit. In some aspects, alternatively or in addition, the light chain of any antibody disclosed herein may further include a light chain constant region (CL), which may be any CL known in the art. In some aspects, CL may be a kappa light chain. In some embodiments, CL may be a lambda light chain. Antibody heavy and light chain constant regions are well known in the art, such as those provided in the IMGT database (www.imgt.org) or at www.vbase2.org / vbstat.php. (both of which are incorporated herein by reference).

[0175] In some aspects, the anti-MYCT1 antibody of the separation herein can be a full-length antibody or an antigen-binding fragment thereof. In some aspects, the antibody of the separation herein can be a full-length antibody, which is an IgG molecule. In some aspects, the antibody of the separation herein can be Fab, (Fab')2 and / or a single-chain antibody. In some aspects, the antibody disclosed herein can be a single-chain antibody (scFv). In some aspects, the scFv antibody herein can include a VH fragment and a VL fragment, which can be connected via a joint. According to these aspects, the joint incorporated between the two variable regions herein can be a flexible joint, a rigid joint, a cleavable joint, or any combination thereof. In some aspects, the joint incorporated between the two variable regions herein can be a flexible peptide joint, a rigid peptide joint, a cleavable peptide joint, or any combination thereof. According to these aspects, the peptide joint incorporated between the two variable regions herein can be at least one amino acid. In some aspects, the peptide linker incorporated between the two variable regions herein can be from about 1 amino acid to about 50 amino acids (e.g., about 1, about 2, about 3, about 4, about 5, about 6, about 7, about 8, about 9, about 10, about 11, about 12, about 13, about 14, about 15, about 16, about 17, about 18, about 19, about 20, about 21, about 22, about 23, about 24, about 25, about 26, about 27, about 28, about 29, about 30, about 32, about 34, about 36, about 38, about 40, about 42, about 44, about 46, about 48, about 50). In some aspects, the scFv antibody herein can comprise a VH fragment and a VL fragment, which can be connected via a flexible peptide linker.

[0176] In some aspects, the scFv antibodies herein can be in a VH-VL orientation (from N-terminus to C-terminus). In some aspects, the scFv antibodies herein can be in a VL-VH orientation (from N-terminus to C-terminus).

[0177] In some respects, the antibody of separation herein can be a human antibody or a humanized antibody. Humanized antibody refers to the form of non-human (e.g., mouse) antibody, which is a specific chimeric immunoglobulin, immunoglobulin chain, or its antigen-binding fragment containing the minimum sequence derived from non-human immunoglobulin. In some respects, the exemplary antibodies described above can each also contain a variant Fc region.

[0178] Polynucleotides, vectors and host cells can be used to prepare any antibody disclosed herein (e.g., anti-MYCT1 antibodies) using recombinant technology, as illustrated herein. In some aspects, nucleic acids (i.e., polynucleotides) encoding the heavy and light chains of antibodies as described herein can be cloned into an expression vector, and each nucleotide sequence is operably connected to a suitable promoter. In some aspects, the polynucleotide sequences encoding the heavy and light chains can each be operably connected to different promoters. In some aspects, the polynucleotide sequences encoding the heavy and light chains can be operably connected to a single promoter so that both the heavy and light chains are expressed from the same promoter. In some embodiments, if necessary, an internal ribosome entry site (IRES) can be inserted between the heavy and light chain coding sequences.

[0179] In certain aspects, the polynucleotide sequences encoding the heavy chains and / or light chains of the antibodies described herein may share one or more of the polynucleotide sequences provided in Table 7.

[0180] In some aspects, the isolated anti-MYCT1 antibody may comprise a heavy chain variable region (VH) comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of any one of SEQ ID NOs: 17, 53, 89, or 125. In some aspects, the anti-MYCT1 antibody comprises a VH comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 17. In some aspects, the anti-MYCT1 antibody comprises a VH comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 53. In some aspects, the anti-MYCT1 antibody comprises a VH comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 89. In some aspects, the anti-MYCT1 antibody comprises a VH comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 125.

[0181] In some aspects, the isolated anti-MYCT1 antibody may comprise a VH comprising a polynucleotide sequence of any one of SEQ ID NOs: 17, 53, 89, or 125. In some aspects, the anti-MYCT1 antibody comprises a VH comprising a polynucleotide sequence of SEQ ID NO: 17. In some aspects, the anti-MY CT1 antibody comprises a VH comprising a polynucleotide sequence of SEQ ID NO: 53. In some aspects, the anti-MYCT1 antibody comprises a VH comprising a polynucleotide sequence of SEQ ID NO: 89. In some aspects, the anti-MYCT1 antibody comprises a VH comprising a polynucleotide sequence of SEQ ID NO: 125.

[0182] In some aspects, the isolated anti-MYCT1 antibody may comprise a light chain variable region (VL) comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of any one of SEQ ID NOs: 35, 71, 107, or 143. In some aspects, the isolated anti-MYCT1 antibody may comprise a VL comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 35. In some aspects, the isolated anti-MYCT1 antibody may comprise a VL comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the sequence of SEQ ID NO: 71. In some aspects, the isolated anti-MYCT1 antibody may comprise a VL comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 107. In some aspects, the isolated anti-MYCT1 antibody may comprise a VL comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 143.

[0183] In some aspects, the isolated anti-MYCT1 antibody may comprise a VL comprising a polynucleotide sequence of any one of SEQ ID NOs: 35, 71, 107, or 143. In some aspects, the isolated anti-MYCT1 antibody may comprise a VL comprising a polynucleotide sequence of SEQ ID NO: 35. In some aspects, the isolated anti-MYCT1 antibody may comprise a VL comprising a polynucleotide sequence of SEQ ID NO: 71. In some aspects, the isolated anti-MYCT1 antibody may comprise a VL comprising a polynucleotide sequence of SEQ ID NO: 107. In some aspects, the isolated anti-MYCT1 antibody may comprise a VL comprising a polynucleotide sequence of SEQ ID NO: 143.

[0184] In some aspects, the isolated anti-MYCT1 antibody may comprise a VH and a VL, wherein the VH comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to a polynucleotide sequence of any one of SEQ ID NOs: 17, 53, 89 or 125; and the VL comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to a polynucleotide sequence of any one of SEQ ID NOs: 35, 71, 107 or 143, or any combination thereof. In some aspects, the isolated anti-MYCT1 antibody may comprise a VH and a VL, wherein the VH comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 17; and the VL comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 35. In some aspects, the isolated anti-MYCT1 antibody may comprise a VH and a VL, wherein the VH comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the sequence of SEQ ID NO: 53; and the VL comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 71. In some aspects, the isolated anti-MYCT1 antibody may comprise a VH and a VL, wherein the VH comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 89; the VL comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 107. In some aspects, the isolated anti-MYCT1 antibody may comprise a VH and a VL, wherein the VH comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 125; the VL comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 143.

[0185] In some aspects, the isolated anti-MYCT1 antibody may comprise a VH and a VL, the VH comprising a polynucleotide sequence of any one of SEQ ID NOs: 17, 53, 89 or 125; the VL comprising a polynucleotide sequence of any one of SEQ ID NOs: 35, 71, 107 or 143, or any combination thereof. In some aspects, the isolated anti-MYCT1 antibody may comprise a VH and a VL, the VH comprising a polynucleotide sequence of SEQ ID NO: 17; the VL comprising a polynucleotide sequence of SEQ ID NO: 35. In some aspects, the isolated anti-MYCT1 antibody may comprise a VH and a VL, the VH comprising a polynucleotide sequence of SEQ ID NO: 53; the VL comprising a polynucleotide sequence of SEQ ID NO: 71. In some aspects, the isolated anti-MYCT1 antibody may comprise a VH and a VL, the VH comprising a polynucleotide sequence of SEQ ID NO: 89; the VL comprising a polynucleotide sequence of SEQ ID NO: 107. In some aspects, the isolated anti-MYCT1 antibody may comprise a VH comprising the polynucleotide sequence of SEQ ID NO: 125 and a VL comprising the polynucleotide sequence of SEQ ID NO: 143.

[0186] In some aspects, the isolated anti-MYCT1 antibody may comprise an H1 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to a polynucleotide sequence of any one of SEQ ID NOs: 9, 45, 81, or 117. In some aspects, the isolated anti-MYCT1 antibody may comprise an H1 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to a polynucleotide sequence of SEQ ID NO: 9. In some aspects, the isolated anti-MYCT1 antibody may comprise an H1 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to a polynucleotide sequence of SEQ ID NO: 45. In some aspects, the isolated anti-MYCT1 antibody may comprise an H1 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 81. In some aspects, the isolated anti-MYCT1 antibody may comprise an H1 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 117.

[0187] In some aspects, the isolated anti-MYCT1 antibody may comprise H1 comprising a polynucleotide sequence of any one of SEQ ID NOs: 9, 45, 81, or 117. In some aspects, the isolated anti-MYCT1 antibody may comprise H1 comprising a polynucleotide sequence of SEQ ID NO: 9. In some aspects, the isolated anti-MYCT1 antibody may comprise H1 comprising a polynucleotide sequence of SEQ ID NO: 45. In some aspects, the isolated anti-MYCT1 antibody may comprise H1 comprising a polynucleotide sequence of SEQ ID NO: 81. In some aspects, the isolated anti-MYCT1 antibody may comprise H1 comprising a polynucleotide sequence of SEQ ID NO: 117.

[0188] In some aspects, the isolated anti-MYCT1 antibody may comprise an L1 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to a polynucleotide sequence of any one of SEQ ID NOs: 27, 63, 99, or 135. In some aspects, the isolated anti-MYCT1 antibody may comprise an L1 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to a polynucleotide sequence of SEQ ID NO: 27. In some aspects, the isolated anti-MYCT1 antibody may comprise an L1 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to a polynucleotide sequence of SEQ ID NO: 63. In some aspects, the isolated anti-MYCT1 antibody may comprise an L1 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 99. In some aspects, the isolated anti-MYCT1 antibody may comprise an L1 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 135.

[0189] In some aspects, the isolated anti-MYCT1 antibody may comprise L1 comprising a polynucleotide sequence of any one of SEQ ID NOs: 27, 63, 99, or 135. In some aspects, the isolated anti-MYC T1 antibody may comprise L1 comprising a polynucleotide sequence of SEQ ID NO: 27. In some aspects, the isolated anti-MYCT1 antibody may comprise L1 comprising a polynucleotide sequence of SEQ ID NO: 63. In some aspects, the isolated anti-MYCT1 antibody may comprise L1 comprising a polynucleotide sequence of SEQ ID NO: 99. In some aspects, the isolated anti-MYCT1 antibody may comprise L1 comprising a polynucleotide sequence of SEQ ID NO: 135.

[0190] In some aspects, the isolated anti-MYCT1 antibody of the present invention may comprise H1 and L1, wherein H1 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to a polynucleotide sequence of any one of SEQ ID NOs: 9, 45, 81 or 117; and L1 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to a polynucleotide sequence of any one of SEQ ID NOs: 27, 63, 99 or 135, or any combination thereof. In some aspects, the isolated anti-MYCT1 antibody herein may comprise H1 and L1, wherein H1 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 9; wherein L1 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 27. In some aspects, the isolated anti-MYCT1 antibody herein may comprise H1 and L1, wherein H1 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 45; wherein L1 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 63. In some aspects, the isolated anti-MYCT1 antibody herein may comprise H1 and L1, wherein H1 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 81; wherein L1 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 99. In some aspects, the isolated anti-MYCT1 antibody herein may comprise H1 and L1, wherein H1 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 117; wherein L1 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 135.

[0191] In some aspects, the isolated anti-MYCT1 antibody herein may comprise H1 and L1, wherein H1 comprises a polynucleotide sequence of any one of SEQ ID NO: 9, 45, 81 or 117; wherein L1 comprises a polynucleotide sequence of any one of SEQ ID NO: 27, 63, 99 or 135, or any combination thereof. In some aspects, the isolated anti-MYCT1 antibody herein may comprise H1 and L1, wherein H1 comprises a polynucleotide sequence of SEQ ID NO: 9; wherein L1 comprises a polynucleotide sequence of SEQ ID NO: 27. In some aspects, the isolated anti-MYCT1 antibody herein may comprise H1 and L1, wherein H1 comprises a polynucleotide sequence of SEQ ID NO: 45; wherein L1 comprises a polynucleotide sequence of SEQ ID NO: 63. In some aspects, the isolated anti-MYCT1 antibody herein may comprise H1 and L1, wherein H1 comprises a polynucleotide sequence of SEQ ID NO: 81; wherein L1 comprises a polynucleotide sequence of SEQ ID NO: 99. In some aspects, the isolated anti-MYCT1 antibody herein may comprise H1 and L1, wherein H1 comprises the polynucleotide sequence of SEQ ID NO: 117; and L1 comprises the polynucleotide sequence of SEQ ID NO: 135.

[0192] In some aspects, the isolated anti-MYCT1 antibody may comprise a heavy chain variable region 2 (H2) comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of any one of SEQ ID NOs: 11, 47, 83, or 119. In some aspects, the isolated anti-MYCT1 antibody may comprise an H2 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 11. In some aspects, the isolated anti-MYCT1 antibody may comprise an H2 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 47. In some aspects, the isolated anti-MYCT1 antibody may comprise an H2 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 83. In some aspects, the isolated anti-MYCT1 antibody may comprise an H2 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 119.

[0193] In some aspects, the isolated anti-MYCT1 antibody may comprise H2 comprising a polynucleotide sequence of any one of SEQ ID NOs: 11, 47, 83, or 119. In some aspects, the isolated anti-MYC T1 antibody may comprise H2 comprising a polynucleotide sequence of SEQ ID NO: 11. In some aspects, the isolated anti-MYCT1 antibody may comprise H2 comprising a polynucleotide sequence of SEQ ID NO: 47. In some aspects, the isolated anti-MYCT1 antibody may comprise H2 comprising a polynucleotide sequence of SEQ ID NO: 83. In some aspects, the isolated anti-MYCT1 antibody may comprise H2 comprising a polynucleotide sequence of SEQ ID NO: 119.

[0194] In some aspects, the isolated anti-MYCT1 antibody may comprise L2, which comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of any one of SEQ ID NOs: 29, 65, 101, or 137. In some aspects, the isolated anti-MYCT1 antibody may comprise L2, which comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 29. In some aspects, the isolated anti-MYCT1 antibody may comprise L2, which comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 65. In some aspects, the isolated anti-MYCT1 antibody may comprise an L2 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 101. In some aspects, the isolated anti-MY CT1 antibody may comprise an L2 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 137.

[0195] In some aspects, the isolated anti-MYCT1 antibody may comprise L2, comprising a polynucleotide sequence of any one of SEQ ID NOs: 29, 65, 101, or 137. In some aspects, the isolated anti-MY CT1 antibody may comprise L2, comprising a polynucleotide sequence of SEQ ID NO: 29. In some aspects, the isolated anti-MYCT1 antibody may comprise L2, comprising a polynucleotide sequence of SEQ ID NO: 65. In some aspects, the isolated anti-MYCT1 antibody may comprise L2, comprising a polynucleotide sequence of SEQ ID NO: 101. In some aspects, the isolated anti-MYCT1 antibody may comprise L2, comprising a polynucleotide sequence of SEQ ID NO: 137.

[0196] In some aspects, the isolated anti-MYCT1 antibody of the present invention may comprise H2 and L2, wherein H2 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to a polynucleotide sequence of any one of SEQ ID NOs: 11, 47, 83 or 119; and L2 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to a polynucleotide sequence of any one of SEQ ID NOs: 29, 65, 101 or 137, or any combination thereof. In some aspects, the isolated anti-MYCT1 antibody herein may include H2 and L2, wherein H2 includes at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 11; and L2 includes at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 29. In some aspects, the isolated anti-MYCT1 antibody herein may include H2 and L2, wherein H2 includes at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 47; and L2 includes at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 65. In some aspects, the isolated anti-MYCT1 antibody herein may include H2 and L2, wherein H2 includes at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 83; and L2 includes at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 101. In some aspects, the isolated anti-MYCT1 antibody herein may include H2 and L2, wherein H2 includes at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 119; and L2 includes at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 137.

[0197] In some aspects, the isolated anti-MYCT1 antibody herein may comprise H2 and L2, wherein H2 comprises a polynucleotide sequence of any one of SEQ ID NO: 11, 47, 83 or 119; wherein L2 comprises a polynucleotide sequence of any one of SEQ ID NO: 29, 65, 101 or 137, or any combination thereof. In some aspects, the isolated anti-MYCT1 antibody herein may comprise H2 and L2, wherein H2 comprises a polynucleotide sequence of SEQ ID NO: 11; wherein L2 comprises a polynucleotide sequence of SEQ ID NO: 29. In some aspects, the isolated anti-MYCT1 antibody herein may comprise H2 and L2, wherein H2 comprises a polynucleotide sequence of SEQ ID NO: 47; wherein L2 comprises a polynucleotide sequence of SEQ ID NO: 65. In some aspects, the isolated anti-MYCT1 antibody herein may comprise H2 and L2, wherein H2 comprises a polynucleotide sequence of SEQ ID NO: 83; wherein L2 comprises a polynucleotide sequence of SEQ ID NO: 101. In some aspects, the isolated anti-MYCT1 antibody herein may comprise H2 and L2, wherein H2 comprises the polynucleotide sequence of SEQ ID NO: 119; and L2 comprises the polynucleotide sequence of SEQ ID NO: 137.

[0198] In some aspects, the isolated anti-MYCT1 antibody may comprise an H3 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of any one of SEQ ID NOs: 13, 49, 85, or 121. In some aspects, the isolated anti-MYCT1 antibody may comprise an H3 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 13. In some aspects, the isolated anti-MYCT1 antibody may comprise an H3 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 49. In some aspects, the isolated anti-MYCT1 antibody may comprise an H3 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 85. In some aspects, the isolated anti-MYCT1 antibody may comprise an H3 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 121.

[0199] In some aspects, the isolated anti-MYCT1 antibody may comprise an H3 comprising a polynucleotide sequence of any one of SEQ ID NOs: 13, 49, 85, or 121. In some aspects, the isolated anti-MYC T1 antibody may comprise an H3 comprising a polynucleotide sequence of SEQ ID NO: 13. In some aspects, the isolated anti-MYCT1 antibody may comprise an H3 comprising a polynucleotide sequence of SEQ ID NO: 49. In some aspects, the isolated anti-MYCT1 antibody may comprise an H3 comprising a polynucleotide sequence of SEQ ID NO: 85. In some aspects, the isolated anti-MYCT1 antibody may comprise an H3 comprising a polynucleotide sequence of SEQ ID NO: 121.

[0200] In some aspects, the isolated anti-MYCT1 antibody may comprise an L3 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of any one of SEQ ID NOs: 31, 67, 103, or 139. In some aspects, the isolated anti-MYCT1 antibody may comprise an L3 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 31. In some aspects, the isolated anti-MYCT1 antibody may comprise an L3 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 67. In some aspects, the isolated anti-MYCT1 antibody may comprise an L3 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 103. In some aspects, the isolated anti-MYCT1 antibody may comprise an L3 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 139.

[0201] In some aspects, the isolated anti-MYCT1 antibody may comprise L3 comprising a polynucleotide sequence of any one of SEQ ID NOs: 31, 67, 103, or 139. In some aspects, the isolated anti-MYCT1 antibody may comprise L3 comprising a polynucleotide sequence of SEQ ID NO: 31. In some aspects, the isolated anti-MYCT1 antibody may comprise L3 comprising a polynucleotide sequence of SEQ ID NO: 67. In some aspects, the isolated anti-MYCT1 antibody may comprise L3 comprising a polynucleotide sequence of SEQ ID NO: 103. In some aspects, the isolated anti-MYCT1 antibody may comprise L3 comprising a polynucleotide sequence of SEQ ID NO: 139.

[0202] In some aspects, the isolated anti-MYCT1 antibody of the present invention may comprise H3 and L3, wherein H3 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to a polynucleotide sequence of any one of SEQ ID NOs: 13, 49, 85 or 121; and L3 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to a polynucleotide sequence of any one of SEQ ID NOs: 31, 67, 103 or 139, or any combination thereof. In some aspects, the isolated anti-MYCT1 antibody herein may include H3 and L3, wherein H3 includes at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 13; wherein L3 includes at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 31. In some aspects, the isolated anti-MYCT1 antibody herein may include H3 and L3, wherein H3 includes at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 49; wherein L3 includes at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 67. In some aspects, the isolated anti-MYCT1 antibody herein may include H3 and L3, wherein H3 includes at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 85; wherein L3 includes at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 103. In some aspects, the isolated anti-MYCT1 antibody herein may include H3 and L3, wherein H3 includes at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 121; wherein L3 includes at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 139.

[0203] In some aspects, the isolated anti-MYCT1 antibody herein may comprise H3 and L3, wherein H3 comprises the polynucleotide sequence of any one of SEQ ID NO: 13, 49, 85 or 121; wherein L3 comprises the polynucleotide sequence of any one of SEQ ID NO: 31, 67, 103 or 139, or any combination thereof. In some aspects, the isolated anti-MYCT1 antibody herein may comprise H3 and L3, wherein H3 comprises the polynucleotide sequence of SEQ ID NO: 13; wherein L3 comprises the polynucleotide sequence of SEQ ID NO: 31. In some aspects, the isolated anti-MYCT1 antibody herein may comprise H3 and L3, wherein H3 comprises the polynucleotide sequence of SEQ ID NO: 49; wherein L3 comprises the polynucleotide sequence of SEQ ID NO: 67. In some aspects, the isolated anti-MYCT1 antibody herein may comprise H3 and L3, wherein H3 comprises the polynucleotide sequence of SEQ ID NO: 85; wherein L3 comprises the polynucleotide sequence of SEQ ID NO: 103. In some aspects, the isolated anti-MYCT1 antibody herein may comprise H3 and L3, wherein H3 comprises the polynucleotide sequence of SEQ ID NO: 121; and L3 comprises the polynucleotide sequence of SEQ ID NO: 139.

[0204] In some aspects, the isolated anti-MYCT1 antibody of the present invention may comprise H1, H2 and H3, wherein H1 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to a polynucleotide sequence of any one of SEQ ID NOs: 9, 45, 81 or 117; the H2 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to a polynucleotide sequence of any one of SEQ ID NOs: 11, 47, 83 or 119; the H3 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to a polynucleotide sequence of any one of SEQ ID NOs: 13, 49, 85 or 121, or any combination thereof. In some aspects, the isolated anti-MYCT1 antibody herein may comprise H1, H2 and H3, wherein H1 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 9; the H2 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 11; the H3 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 13, or any combination thereof. In some aspects, the isolated anti-MYCT1 antibody of the present invention may comprise H1, H2 and H3, wherein H1 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO:45; the H2 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO:47; the H3 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO:49, or any combination thereof. In some aspects, the isolated anti-MYCT1 antibody of the present invention may comprise H1, H2 and H3, wherein H1 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 81; the H2 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 83; the H3 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 85, or any combination thereof.In some aspects, the isolated anti-MYCT1 antibody of the present invention may comprise H1, H2 and H3, wherein H1 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 117; the H2 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 119; the H3 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 121, or any combination thereof.

[0205] In some aspects, the isolated anti-MYCT1 antibody herein may include H1, H2, and H3, wherein H1 includes a polynucleotide sequence of any one of SEQ ID NO: 9, 45, 81, or 117; H2 includes a polynucleotide sequence of any one of SEQ ID NO: 11, 47, 83, or 119; H3 includes a polynucleotide sequence of any one of SEQ ID NO: 13, 49, 85, or 121, or any combination thereof. In some aspects, the isolated anti-MYCT1 antibody herein may include H1, H2, and H3, wherein H1 includes a polynucleotide sequence of SEQ ID NO: 9; H2 includes a polynucleotide sequence of SEQ ID NO: 11; H3 includes a polynucleotide sequence of SEQ ID NO: 13, or any combination thereof. In some aspects, the isolated anti-MYCT1 antibody herein may include H1, H2, and H3, wherein H1 includes a polynucleotide sequence of SEQ ID NO: 45; H2 includes a sequence of SEQ ID NO: 47; H3 includes a polynucleotide sequence of SEQ ID NO: 49, or any combination thereof. In some aspects, the isolated anti-MYC T1 antibody herein may comprise H1, H2 and H3, wherein H1 comprises the polynucleotide sequence of SEQ ID NO: 81; H2 comprises the polynucleotide sequence of SEQ ID NO: 83; H3 comprises the polynucleotide sequence of SEQ ID NO: 85, or any combination thereof. In some aspects, the isolated anti-MYCT1 antibody herein may comprise H1, H2 and H3, wherein H1 comprises the polynucleotide sequence of SEQ ID NO: 117; H2 comprises the polynucleotide sequence of SEQ ID NO: 119; H3 comprises the polynucleotide sequence of SEQ ID NO: 121, or any combination thereof.

[0206] In some aspects, the isolated anti-MYCT1 antibody of the present invention may comprise L1, L2 and L3, wherein L1 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to a polynucleotide sequence of any one of SEQ ID NOs: 27, 63, 99 or 135; the L2 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to a polynucleotide sequence of any one of SEQ ID NOs: 29, 65, 101 or 137; the L3 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to a polynucleotide sequence of any one of SEQ ID NOs: 31, 67, 103 or 139, or any combination thereof. In some aspects, the isolated anti-MYCT1 antibody herein may comprise L1, L2 and L3, wherein L1 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 27; the L2 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 29; the L3 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 31, or any combination thereof. In some aspects, the isolated anti-MYCT1 antibody of the present invention may comprise L1, L2 and L3, wherein L1 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 63; the L2 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 65; the L3 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 67, or any combination thereof. In some aspects, the isolated anti-MYCT1 antibody of the present invention may comprise L1, L2 and L3, wherein L1 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 99; the L2 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 101; the L3 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 103, or any combination thereof.In some aspects, the isolated anti-MYCT1 antibody herein may comprise L1, L2 and L3, wherein L1 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 135; the L2 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 137; the L3 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 139, or any combination thereof.

[0207] In some aspects, the isolated anti-MYCT1 antibody herein may comprise L1, L2 and L3, wherein L1 comprises a polynucleotide sequence of any one of SEQ ID NO: 27, 63, 99 or 135; wherein L2 comprises a polynucleotide sequence of any one of SEQ ID NO: 29, 65, 101 or 137; wherein L3 comprises a polynucleotide sequence of any one of SEQ ID NO: 31, 67, 103 or 139, or any combination thereof. In some aspects, the isolated anti-MYCT1 antibody herein may comprise L1, L2 and L3, wherein L1 comprises a polynucleotide sequence of SEQ ID NO: 27; wherein L2 comprises a polynucleotide sequence of SEQ ID NO: 29; wherein L3 comprises a polynucleotide sequence of SEQ ID NO: 31, or any combination thereof. In some aspects, the isolated anti-MYCT1 antibody herein may include L1, L2, and L3, wherein L1 includes the polynucleotide sequence of SEQ ID NO: 63; L2 includes the polynucleotide sequence of SEQ ID NO: 65; L3 includes the polynucleotide sequence of SEQ ID NO: 67, or any combination thereof. In some aspects, the isolated anti-MYCT1 antibody herein may include L1, L2, and L3, wherein L1 includes the polynucleotide sequence of SEQ ID NO: 99; L2 includes the polynucleotide sequence of SEQ ID NO: 101; L3 includes the polynucleotide sequence of SEQ ID NO: 103, or any combination thereof. In some aspects, the isolated anti-MYCT1 antibody herein may include L1, L2, and L3, wherein L1 includes the polynucleotide sequence of SEQ ID NO: 135; L2 includes the polynucleotide sequence of SEQ ID NO: 137; L3 includes the polynucleotide sequence of SEQ ID NO: 139, or any combination thereof.

[0208] In some aspects, the isolated anti-MYCT1 antibody herein may comprise H1, H2, H3, L1, L2 and L3, wherein H1 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to a polynucleotide sequence of any one of SEQ ID NOs: 9, 45, 81 or 117; the H2 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to a polynucleotide sequence of any one of SEQ ID NOs: 11, 47, 83 or 119; the H3 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to a polynucleotide sequence of any one of SEQ ID NOs: 13, 49, 85 or 121; the L1 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to a polynucleotide sequence of any one of SEQ ID NOs: 121; NO:27, 63, 99 or 135, any one of the polynucleotide sequence of at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity; the L2 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity with the polynucleotide sequence of any one of SEQ ID NO:29, 65, 101 or 137; the L3 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity with the polynucleotide sequence of any one of SEQ ID NO:31, 67, 103 or 139, or any combination thereof. In some aspects, the isolated anti-MYCT1 antibody herein may comprise H1, H2, H3, L1, L2 and L3, wherein H1 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 9; the H2 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 11; the H3 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of any one of SEQ ID NO: 13; the L1 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 27; the L2 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 29; at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO:29; said L3 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO:31, or any combination thereof.In some aspects, the isolated anti-MYCT1 antibody herein may comprise H1, H2, H3, L1, L2 and L3, wherein H1 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 45; the H2 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 47; the H3 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of any one of SEQ ID NO: 49; the L1 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 63; the L2 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 64; NO:65; said L3 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity with the polynucleotide sequence of SEQ ID NO:67, or any combination thereof. In some aspects, the isolated anti-MYCT1 antibody herein may comprise H1, H2, H3, L1, L2 and L3, wherein H1 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 81; the H2 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 83; the H3 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of any one of SEQ ID NO: 85; the L1 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the sequence of SEQ ID NO: 99; the L2 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 99; NO:101 has at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity with the polynucleotide sequence of NO:101; the L3 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity with the polynucleotide sequence of SEQ ID NO:103, or any combination thereof.In some aspects, the isolated anti-MYCT1 antibody herein may comprise H1, H2, H3, L1, L2 and L3, wherein H1 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 117; the H2 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 119; the H3 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of any one of SEQ ID NO: 121; the L1 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 135; the L2 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 137; The L3 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity with the polynucleotide sequence of SEQ ID NO: 137; the L3 comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity with the polynucleotide sequence of SEQ ID NO: 139, or any combination thereof.

[0209] In some aspects, the isolated anti-MYCT1 antibody herein may comprise H1, H2, H3, L1, L2 and L3, wherein H1 comprises a polynucleotide sequence of any one of SEQ ID NO: 9, 45, 81 or 117; H2 comprises a sequence of any one of SEQ ID NO: 11, 47, 83 or 119; H3 comprises a polynucleotide sequence of any one of SEQ ID NO: 13, 49, 85 or 121; L1 comprises a polynucleotide sequence of any one of SEQ ID NO: 27, 63, 99 or 135; L2 comprises a polynucleotide sequence of any one of SEQ ID NO: 29, 65, 101 or 137; L3 comprises a polynucleotide sequence of any one of SEQ ID NO: 31, 67, 103 or 139, or any combination thereof. In some aspects, the isolated anti-MYCT1 antibody herein may include H1, H2, H3, L1, L2 and L3, wherein H1 includes the polynucleotide sequence of SEQ ID NO: 9; H2 includes the sequence of SEQ ID NO: 11; H3 includes the polynucleotide sequence of any one of SEQ ID NO: 13; L1 includes the polynucleotide sequence of SEQ ID NO: 27; L2 includes the sequence of SEQ ID NO: 29; L3 includes the polynucleotide sequence of SEQ ID NO: 31, or any combination thereof. In some aspects, the isolated anti-MYCT1 antibody herein may include H1, H2, H3, L1, L2 and L3, wherein H1 includes the polynucleotide sequence of SEQ ID NO: 45; H2 includes the polynucleotide sequence of SEQ ID NO: 47; H3 includes the sequence of any one of SEQ ID NO: 49; L1 includes the polynucleotide sequence of SEQ ID NO: 63; L2 includes the sequence of SEQ ID NO: 65; L3 includes the polynucleotide sequence of SEQ ID NO: 67, or any combination thereof. In some aspects, the isolated anti-MYCT1 antibody herein may comprise H1, H2, H3, L1, L2 and L3, wherein H1 comprises the polynucleotide sequence of SEQ ID NO:81; H2 comprises the polynucleotide sequence of SEQ ID NO:83; H3 comprises the polynucleotide sequence of any one of SEQ ID NO:85; L1 comprises the polynucleotide sequence of SEQ ID NO:99; L2 comprises the polynucleotide sequence of SEQ ID NO:101; L3 comprises the polynucleotide sequence of SEQ ID NO:103, or any combination thereof.In some aspects, the isolated anti-MYCT1 antibody herein may comprise H1, H2, H3, L1, L2 and L3, wherein H1 comprises the polynucleotide sequence of SEQ ID NO: 117; H2 comprises the polynucleotide sequence of SEQ ID NO: 119; H3 comprises the polynucleotide sequence of any one of SEQ ID NO: 121; L1 comprises the polynucleotide sequence of SEQ ID NO: 135; L2 comprises the polynucleotide sequence of SEQID NO: 137; L3 comprises the polynucleotide sequence of SEQ ID NO: 139, or any combination thereof.

[0210] In certain aspects, the expression and / or production of antibodies in host cells can be promoted by expressing a leader peptide in front of the variable region of the heavy or light chain of the antibody. In some embodiments, the leader peptide can be optimized for expression in certain host cells (e.g., E. coli).

[0211] In some respects, genetically engineered antibodies such as single-chain antibodies can be produced via, for example, conventional recombinant techniques or any method known in the art. In some respects, the DNA encoding the monoclonal antibody specific to the target antigen can be easily separated and sequenced using conventional procedures (for example, by using oligonucleotide probes that can specifically bind the heavy chain and light chain of the encoding monoclonal antibody). Once separated, DNA can be placed in one or more expression vectors, then transfected into a host cell that does not produce immunoglobulin in addition, the host cell such as Escherichia coli cells, monkey COS cells, Chinese hamster ovary (CHO) cells or myeloma cells, to obtain the synthesis of monoclonal antibodies in recombinant host cells. DNA can then be modified, for example, by replacing homologous mouse sequences with the encoding sequences of people's heavy chain and light chain constant regions, or by covalently linking the encoding sequences of non-immunoglobulin polypeptides, in whole or in part, to immunoglobulin coding sequences. In some respects, genetically engineered antibodies with target antigen binding specificity can be prepared, for example, chimeric or hybrid antibodies.

[0212] In some aspects, the single-chain antibody herein can be prepared by connecting the nucleotide sequence encoding the heavy chain variable region and the nucleotide sequence encoding the light chain variable region via recombinant technology. In some aspects, a joint can be incorporated between the two variable regions. In some embodiments, the described technology for producing single-chain antibodies can be applied to produce phage or yeast scFv libraries, and scFv clones specific to MY CT1 can be identified from the library following conventional procedures. In some embodiments, positive clones can be subjected to further screening to identify those that bind to specific epitopes on MYCT1.

[0213] In some respects, one or more vectors (e.g., expression vectors) with nucleic acids encoding any antibody herein can be introduced into suitable host cells to produce antibodies. In some respects, host cells can be cultured under conditions suitable for expressing antibodies or any of their polypeptide chains. In some respects, antibodies or their polypeptide chains can be recovered by cultivated cells (e.g., from cells or culture supernatants) via conventional methods, such as affinity purification of the conventional methods. In some respects, the polypeptide chains of the antibodies herein can be incubated in suitable time periods under suitable conditions, allowing the production of antibodies.

[0214] In some aspects, the method for preparing the antibody described herein may include a recombinant expression vector encoding the heavy chain and light chain of the antibody in combination with MYCT1, also as described herein. In some aspects, the recombinant expression vector may be introduced into a suitable host cell (e.g., dhfr-CHO cell) by conventional methods, such as calcium phosphate-mediated transfection. In some aspects, positive transformant host cells may be selected and cultured under suitable conditions for allowing the expression of two polypeptide chains forming the antibody, which may be recovered from the cell or from the culture medium. In some aspects, the two chains recovered from the host cell may be incubated under suitable conditions for allowing the formation of the antibody.

[0215] In some aspects, two recombinant expression vectors can be provided, one encoding the heavy chain of the disclosed antibody, and the other encoding the light chain of the disclosed antibody. In some aspects, both of the two recombinant expression vectors can be introduced into suitable host cells (such as dhfr-CHO cells) by conventional methods, and the conventional methods are such as calcium phosphate-mediated transfection. In some aspects, the expression vector can be introduced into suitable host cells separately. In some aspects, positive transformants can be selected and cultivated under the suitable conditions of allowing the polypeptide chain expression of the antibody. In some aspects, when two expression vectors are introduced into the same host cell, the antibody wherein produced can be recovered from the host cell or from the culture medium. In some aspects, the polypeptide chain can be recovered from the host cell or from the culture medium, and then hatched under the suitable conditions of allowing the formation of the antibody. In some aspects, when two expression vectors are introduced into different host cells, they can be recovered from the corresponding host cell or from the corresponding culture medium separately. In some embodiments, then two polypeptide chains can be hatched under the conditions suitable for forming an antibody.

[0216] In certain aspects, standard molecular biology techniques can be used to prepare recombinant expression vectors, transfect host cells, select transformants, culture host cells and recover antibodies from the culture medium. In some embodiments, some antibodies can be separated by affinity chromatography using protein A or protein G coupled matrix.

[0217] In other aspects, the antibodies herein can be characterized by identifying an epitope or more than one epitope to which an antigen binds or "epitope mapping". There are many methods known in the art for mapping and characterizing the location of epitopes on proteins, including, but not limited to, solving crystal structures of antibody-antigen complexes, competition assays, gene fragment expression assays, and synthetic peptide-based assays. In some embodiments, epitope mapping can be used to determine the sequence to which an antibody binds.

[0218] Chimeric Antigen Receptor

[0219] In some aspects, the disclosure encompasses chimeric antigen receptor (CAR) immune effector cells (CAR- immune effector cells) comprising antigen binding domains, the antigen binding domains comprising antibodies or fragments thereof specifically binding to MYCT1 disclosed herein. In one aspect, the CAR- immune effector cells comprising antigen binding domains comprising antibodies or fragments thereof specifically binding to MYCT1 include amino acids or polynucleotide sequences listed in Table 7.

[0220] The hinge domain is a structure between the antigen recognition domain (e.g., an antibody that binds MYCT1) and the cytoplasmic membrane; these sequences are typically derived from IgG subclasses (e.g., IgG1 and IgG4), IgD, and CD8 domains, with IgG1 being the most widely used for CAR construction. Currently, research on the hinge domain focuses on the following four aspects: 1) reducing binding affinity to Fcγ receptors, thereby eliminating off-target activation; 2) enhancing the flexibility of the single-chain variable fragment (scFv), thereby relieving the spatial restrictions between tumor antigens and CARs, and subsequently promoting synaptic formation between CAR-immune effector cells and target cells; 3) reducing the distance between scFv and the target epitope; and 4) using anti-Fc reagents to facilitate detection of CAR expression.

[0221] In one aspect, the hinge domain comprises a hinge domain of a human protein selected from the group consisting of CD28, 4-1BB (CD137), OX-40 (CD134), CD3ζ, T cell receptor α or β chain, CD45, CD4, CD 5, CD8, CD8α, CD9, CD16, CD22, CD33, CD37, CD64, CD80, CD86, ICOS, CD154, functional derivatives thereof, and / or combinations thereof.

[0222] Typically, the extracellular domain (i.e., antigen recognition domain (e.g., antibody binding to MYCT1)) or targeting element is connected to the intracellular domain via a transmembrane domain, which is the co-stimulatory and signaling domain of the chimeric antigen receptor. The transmembrane domain crosses the cell membrane, anchors the CAR to the surface of the immune effector cell, and connects the extracellular domain to the intracellular domain, thereby affecting the expression of CAR on the surface of the immune effector cell.

[0223] Transmembrane domains include hydrophobic polypeptides that span the cell membrane. In particular, transmembrane domains span from one side of the cell membrane (extracellular) to the other side of the cell membrane (intracellular or cytoplasmic).

[0224] The transmembrane domain may be in the form of an alpha helix or a beta barrel or a combination thereof. The transmembrane domain may include a polytopic protein having many transmembrane segments, each of which is an alpha-helix, a beta sheet or a combination thereof.

[0225] In one aspect, a transmembrane domain naturally associated with a domain in CAR is used. In another aspect, a transmembrane domain is selected or modified by amino acid substitution to avoid binding of such domains to transmembrane domains of the same or different surface membrane proteins to minimize interactions with other members of the receptor complex.

[0226] For example, the transmembrane domain is selected from the following transmembrane domains: T cell receptor α or β chain, CD3 ζ chain, CD28, CD3s, CD45, CD4, CD5, CD7, CD8, CD9, CD16, CD22, CD33, CD37, CD64, CD80, CD86, CD68, OX-40 (CD134), 4-1BB (CD137), ICOS, CD41, CD154, their functional derivatives and / or combinations.

[0227] The chimeric antigen receptors of the present disclosure may comprise one or more co-stimulatory domains and / or one or more spacers. The co-stimulatory domains are derived from co-stimulatory proteins that enhance cytokine production, proliferation, cytotoxicity and / or persistence in vivo.

[0228] In one aspect, the co-stimulatory domain is selected from OX-40 (CD134), CD27, CD28, CD30, CD40, PD-1, CD2, CD7, CD258, natural killer cell group 2 member C (NKG2C), natural killer cell group 2 member D (NKG2D), B7-H3, a ligand that binds at least one of the following: CD83, ICAM-1, LFA-1 (CD11a / CD18), ICOS and 4-1BB (CD137), CDS, ICAM-1, LFA-1 (CD1a / CD18), CD40, CD27, active fragments thereof, functional derivatives thereof, and combinations thereof.

[0229] The chimeric antigen receptor of the present disclosure also includes a signaling domain, which provides an intracellular signal to the immune effector cell when the antigen binds to the antigen recognition domain. The signaling domain of the chimeric antigen receptor of the present disclosure is responsible for activating at least one of the effector functions of the immune effector cell in which the chimeric receptor is expressed. The term "effector function" refers to the specialized function of a differentiated cell. The effector function of an immune cell may be, for example, a cytolytic activity or an auxiliary activity including the secretion of a cytokine. The effector function in naive, memory or memory immune cells may also include antigen-dependent proliferation. Therefore, the term "intracellular domain" refers to the intracellular portion of a CAR, which transduces effector function signals after antigen binding to the extracellular domain and instructs immune cells to perform specialized functions. Non-limiting examples of suitable signaling domains include the ζ chain of an immune cell receptor or any homolog thereof (e.g., η, δ, γ, or ε), MB-1 chain, 829, FcRIII, FcRI, and combinations of signaling molecules, such as CD3ζ and CD28, CD27, 4-1BB (CD137), DNAX-activating protein 10 (DAP10), OX-40 (CD134), and combinations thereof, as well as other similar molecules and fragments. Signaling domains of other activating proteins, such as FcγRIII and Fc€RI, can be used. Although the entire signaling domain is generally used, in many cases, it is not necessary to use the entire intracellular polypeptide. To that extent, a truncated portion of the signaling domain can be used as long as it still transduces the effector function signal. The term intracellular domain also means a truncated portion of any intracellular domain that is sufficient to transduce an effector function signal. Alternatively or in addition, the CAR-immune effector cells included in the present disclosure may further include one or more suicide genes. As used herein, "suicide gene" refers to a nucleic acid sequence introduced into CAR- immune effector cells by standard methods known in the art, which, when activated, causes the death of CAR- immune effector cells. If necessary, suicide genes can promote effective tracking and elimination of CAR- immune effector cells in vivo. Killing promoted by activating suicide genes can occur by methods known in the art. Suitable suicide gene therapy systems known in the art include, but are not limited to, various herpes simplex virus thymidine kinase (HSVtk) / ganciclovir (GCV) suicide gene therapy systems or inducible caspase 9 proteins. In an exemplary embodiment, the suicide gene is a CD34 / thymidine kinase chimeric suicide gene.

[0230] Methods for CAR design, delivery and expression in immune cells and the manufacture of clinical-grade CAR-immune effector cell groups are known in the art. For example, see Lee et al., Clin. Cancer Res., 2012, 18 (10): 2780-90, which is incorporated herein by reference in its entirety. For example, a retrovirus can be used to introduce engineered CAR into immune effector cells, and the retrovirus effectively and stably integrates the nucleic acid sequence encoding the chimeric antigen receptor into the target cell genome. An exemplary method for viral vector production is described in the method to the embodiment. Other methods known in the art include, but are not limited to, lentiviral transduction, transposon-based systems, direct RNA transfection, and CRISPR / Cas systems (e.g., type I, type II, or type III systems, which use suitable Cas proteins, such as Cas3, Cas4, Cas5, Cas5e (or CasD), Cas6, Cas6e, Cas6f, Cas7, Cas8a1, Cas8a2, Cas8b, Cas8c, Cas9, Cas10, Cas1 Od, CasF, CasG, Cas , Csb1, Csb2, Csb3, Csx17, Csx14, Csx10, Csx16, CsaX, Csx3, Csz1, Csx15, Csf1, Csf2, Csf3, Csf4, and Cu1966, etc.).

[0231] In some aspects, the immune effector cell is selected from the group consisting of a T cell, a natural killer (NK) cell, a cytotoxic T lymphocyte (CTL), and a regulatory T cell.

[0232] In some aspects, immune effector cells include any leukocytes involved in protecting the body from infectious diseases and foreign substances. For example, immune effector cells can include lymphocytes, monocytes, macrophages, dendritic cells, mast cells, neutrophils, basophils, eosinophils or any combination thereof. In some aspects, immune effector cells include T lymphocytes.

[0233] As disclosed herein, the T cell can be any subset of T cells, non-limiting examples of which can include T helper cells (TH cells), cytotoxic T cells (TC cells or CTLs), memory T cells, regulatory T cells (Treg cells), or natural killer T (NKT) cells.

[0234] T helper cells (TH cells) assist other leukocytes in the immune process, including B cell maturation into plasma cells and memory B cells, and the activation of cytotoxic T cells and macrophages. These cells are also referred to as CD4+T cells because they express CD4 glycoproteins on their surfaces. When MHC class II molecules present peptide antigens to helper T cells, the helper T cells become activated, and the MHC class II molecules are expressed on the surface of antigen presenting cells (APC). Once activated, they rapidly divide and secrete small proteins called cytokines, which regulate or assist active immune responses. These cells can be differentiated into one of several subtypes, including TH1, TH2, TH3, TH17, TH9 or TFH, which secrete different cytokines to promote different types of immune responses.

[0235] Cytotoxic T cells (TC cells or CTLs) destroy virus-infected cells and tumor cells, and are also involved in transplant rejection. These cells are also referred to as CD8+T cells because they express CD8 glycoproteins on their surfaces. These cells recognize their targets by binding to antigens associated with MHC class I molecules, which are present on the surface of all nucleated cells. Through IL-10, adenosine and other molecules secreted by regulatory T cells, CD8+ cells can be inactivated to an anergic state, which prevents autoimmune diseases.

[0236] Memory T cells are a subset of antigen-specific T cells that persist long after the infection resolves. They rapidly expand into large numbers of effector T cells upon re-exposure to their cognate antigens, thus providing the immune system with a "memory" of past infections. Memory cells can be CD4+ or CD8+. Memory T cells typically express the cell surface protein CD45RO.

[0237] Regulatory T cells (Treg cells), formerly known as suppressor T cells, are crucial for maintaining immune tolerance. Their main role is to shut down T cell-mediated immunity to the end of the immune response and to suppress autoreactive T cells that escape the negative selection process in the thymus. Two main types of CD4+Treg cells are described - naturally occurring Treg cells and adaptive Treg cells.

[0238] Natural killer T (NKT) cells connect the adaptive immune system with the innate immune system, and the natural killer T (NKT) cells should not be confused with natural killer (NK) cells. Unlike conventional T cells that recognize peptide antigens presented by major histocompatibility complex (MH C) molecules, NKT cells recognize glycolipid antigens presented by MHC class I molecules (MH C like I) CD1d. In some aspects, NKT cells are constant natural killer T (iNKT) cells. iNKT cells recognize CD1d and are restricted by their T cell receptors (TCRs), which are Vα24Jα18 in humans and are typically paired with Vβ11. iNKT cells further reduce graft-versus-host disease (GVHD).

[0239] In some aspects, T cells comprise a mixture of CD4+ cells. In other aspects, T cells are enriched in one or more subsets based on cell surface expression. For example, in some cases, T cells comprise cytotoxic CD8+ T lymphocytes. In some embodiments, T cells comprise γδT cells, which have a unique T cell receptor (TCR) with a γ chain and a δ chain instead of α and β chains.

[0240] Natural killer (NK) cells are CD56+CD3- large granular lymphocytes that can kill virus-infected and transformed cells and constitute a key cell subset of the innate immune system. Unlike cytotoxic CD8+ T lymphocytes, NK cells initiate cytotoxicity against tumor cells without the need for prior sensitization and can also eradicate MHC-I-negative cells.

[0241] In some aspects, immune effector cells can be obtained from the subject to be treated (i.e., autologous), and can be applied after genetic modification to express a CAR with a disclosed anti-MYCT1 antigen recognition domain (e.g., an anti-MYCT1 antibody or fragment thereof disclosed herein), as an adoptive transfer. In this regard, adoptive transfer comprises obtaining the immune effector cells of the subject, and then genetically modifying the cells to express a disclosed CAR with an anti-MYCT1 antigen recognition domain, and infusing the modified cells back into the subject. In some aspects, immune effector cell lines or donor effector cells (allogeneic) are used. Immune effector cells for allogeneic therapy can be collected from a single subject or multiple subjects. Immune effector cells can be obtained from a variety of sources, including peripheral blood mononuclear cells, bone marrow, lymph node tissue, umbilical cord blood, thymus tissue, tissue from the site of infection, ascites, pleural effusion, spleen tissue and tumor. Immune effector cells can be obtained from blood collected from subjects using any number of techniques known to professionals, such as Ficoll TMFor example, cells from an individual's circulating blood can be obtained by apheresis. In some aspects, immune effector cells can be isolated from peripheral blood lymphocytes by lysing red blood cells and depleting monocytes, for example, by PERCOLL TM Gradient centrifugation or by countercurrent centrifugal elutriation. Specific subpopulations of immune effector cells can be further separated by positive or negative selection techniques. For example, immune effector cells can be separated using a combination of antibodies for surface markers unique to positively selected cells, such as by incubating with antibody-conjugated beads for a period of time sufficient to positively select the desired immune effector cells. Alternatively, the enrichment of immune effector cell populations can be achieved by negative selection using a combination of antibodies for surface markers unique to negatively selected cells.

[0242] In some aspects, the expression of the nucleic acid encoding CAR can be operably connected to a promoter by encoding a nucleic acid comprising a disclosed anti-MYCT 1 antigen recognition domain of a CAR polypeptide, and the construct is incorporated into an expression vector to achieve. Typical cloning vectors contain transcription and translation terminators, initiation sequences, and promoters that can be used to regulate the expression of desired nucleic acid sequences. In some aspects, immune effector cells further include modifications of endogenous T cell receptor alpha chains (TRAC) so that endogenous T cell receptor-mediated signal transduction is blocked in CAR-T cells.

[0243] In some aspects, a CAR comprising a disclosed anti-MYCT1 antigen recognition domain can be cloned into various types of vectors. For example, a CAR comprising a disclosed anti-MYCT1 antigen recognition domain can be cloned into a vector, including but not limited to a plasmid, a phagemid, a phage derivative, an animal virus, and a cosmid. In some aspects, non-limiting examples of vectors include expression vectors, replication vectors, probe generation vectors, and sequencing vectors.

[0244] Many virus-based systems have been developed for gene transfer into mammalian cells. For example, a CAR comprising a disclosed anti-MYCT1 antigen recognition domain can be inserted into a vector and packaged in a retroviral particle using techniques known in the art. The recombinant virus can then be isolated and delivered to the subject's cells in vivo or ex vivo.

[0245] Bispecific, multispecific or hybrid antibodies

[0246] In some aspects, the present disclosure further encompasses bispecific antibodies, multispecific antibodies, or hybrid antibodies having at least two different binding domains with different specificities, wherein at least one domain comprises H1, H2, H3, L1, L2, L3, VH, and / or VL disclosed in Table 7. In some aspects, the bispecific antibody, multispecific antibody, or hybrid antibody comprises anti-MYCT1 H1, H2, H3, L1, L2, L3, VH, and / or VL disclosed herein and at least one immune cell engager (e.g., T cell engager, NK cell engager, iNKT cell engager, B cell engager, dendritic cell engager, or macrophage engager). Such antibody constructs are recombinant protein constructs made of two flexibly linked antibody-derived binding domains, wherein one binding domain is specific for a selected tumor antigen on a target tumor cell, and the second binding domain binds and / or activates T cells, NK cells, iNKT cells, B cells, dendritic cells, or macrophages. In some aspects, the disclosed antibodies can transiently guide immune cells to target tumor cells and / or activate the inherent cytolytic potential of immune cells against target tumor cells. In some aspects, bispecific antibodies, multispecific antibodies or hybrid antibodies comprise at least one domain comprising anti-MYCT1 H1, H2, H3, L1, L2, L3, VH and / or VL disclosed herein, and a second domain comprising a binding domain specific for an antigen expressed by a B cell and / or plasma cell tumor cell. Bispecific antibodies, multispecific antibodies or hybrid antibody constructs can be produced by a variety of methods, including hybridoma fusion, Fab' fragment connection or any other method known in the art.

[0247] In some aspects, a bispecific antibody, a multispecific antibody, or a hybrid antibody comprises a T cell engager (e.g., a bispecific T cell engager (BiTE)) comprising a first antigen recognition domain and a second antigen recognition domain, wherein the first antigen recognition domain comprises an anti-MYCT1 H1, H2, H3, L1, L2, L3, VH, and / or VL disclosed herein, and the second antigen recognition domain comprises an antibody or antigen-binding antibody fragment that binds or activates T cells. In this regard, the second antigen recognition domain comprising an antibody or antigen-binding antibody fragment binds to one or more of the following: CD2, CD3, CD4, CD5, CD6, CD8, CD25, CD27, CD28, CD30, CD40, CD40L, CD44, CD45, CD69, CD90, CRa, TCRp, TCRy, TCRC, ICOS, HVEM, LIGHT, 4-1BB, OX40, DR3, GITR, TIM1, SLAM, or CD226.

[0248] In some aspects, the bispecific antibody, multispecific antibody or hybrid antibody has a binding domain that engages NK cells. In this regard, the bispecific antibody comprises a first antigen recognition domain and a second antigen recognition domain, wherein the first antigen recognition domain comprises an anti-MYCT1 H1, H2, H3, L1, L2, L3, VH and / or VL disclosed herein, and the second antigen recognition domain comprises an antibody or antigen-binding antibody fragment that binds or activates NK cells. In this regard, the second antigen recognition domain comprising the antibody or antigen-binding antibody fragment binds one or more of the following: CD16 (e.g., CD16a, CD16b, or both), NKp30, NKp40, NKp44, NKp46, NKG2D, DNAM1, DAP10, CRTAM, CD27, PSGL1, CD96, CD100 (SEMA4D), NKp80, CD244 (also known as SLAMF4 or 2B4), SLAMF6, SLAMF7, KIR2DS2, KIR2DS4, KIR3DS1, KIR2DS3, KIR2DS5, KIR2DS1, CD94, NKG2C, NKG2E, or CD160.

[0249] In some aspects, bispecific antibodies, multispecific antibodies or hybrid antibodies have a binding domain that engages iNKT cells. In this regard, the bispecific antibody comprises a first antigen recognition domain and a second antigen recognition domain, wherein the first antigen recognition domain comprises anti-MYCT1 H1, H2, H3, L1, L2, L3, VH and / or VL disclosed herein, and the second antigen recognition domain comprises an antibody or antigen-binding antibody fragment that binds or activates iNKT cells. In this regard, the second antigen recognition domain comprising an antibody or antigen-binding antibody fragment binds to CD3.

[0250] In some aspects, bispecific antibodies, multispecific antibodies or hybrid antibodies have a binding domain that engages B cells. In this regard, the bispecific antibody comprises a first antigen recognition domain and a second antigen recognition domain, wherein the first antigen recognition domain comprises anti-MYCT1 H1, H2, H3, L1, L2, L3, VH and / or VL disclosed herein, and the second antigen recognition domain comprises an antibody or antigen-binding antibody fragment that binds or activates B cells. In this regard, the second antigen recognition domain comprising an antibody or antigen-binding antibody fragment binds to one or more of OX40, CD40 or CD70.

[0251] In some aspects, bispecific antibodies, multispecific antibodies or hybrid antibodies have a binding domain that engages dendritic cells. In this regard, the bispecific antibody comprises a first antigen recognition domain and a second antigen recognition domain, wherein the first antigen recognition domain comprises anti-MYCT1 H1, H2, H3, L1, L2, L3, VH and / or VL disclosed herein, and the second antigen recognition domain comprises an antibody or antigen-binding antibody fragment that binds or activates dendritic cells. In this regard, the second antigen recognition domain comprising an antibody or antigen-binding antibody fragment binds to one or more of CD2, OX40, 4-1BB, TLR4, CD47 or STING.

[0252] In some aspects, bispecific antibodies, multispecific antibodies or hybrid antibodies have a binding domain that engages macrophages. In this regard, the bispecific antibody comprises a first antigen recognition domain and a second antigen recognition domain, wherein the first antigen recognition domain comprises anti-MYCT1 H1, H2, H3, L1, L2, L3, VH and / or VL disclosed herein, and the second antigen recognition domain comprises an antibody or antigen-binding antibody fragment that binds or activates macrophages. In this regard, the second antigen recognition domain comprising an antibody or antigen-binding antibody fragment binds to one or more of OX40, CD40, CD70, TLR4, TLR9, CD47 or STING agonists.

[0253] In some aspects, the bispecific antibody, multispecific antibody or hybrid antibody comprises a first antigen recognition domain and a second antigen recognition domain, wherein the first antigen recognition domain comprises anti-MY CT1 H1, H2, H3, L1, L2, L3, VH and / or VL disclosed herein, and the second antigen recognition domain comprises an antibody or antigen-binding antibody fragment that binds to an antigen expressed by a tumor cell. In some aspects, the antigen is an antigen expressed by a sarcoma and / or breast cancer. In this regard, the second antigen recognition domain comprising an antibody or an antigen-binding antibody fragment binds to one or more of the following: MAGE-A4, NY-ESO-1, PRAME, TR AG-3 / CSAGE, SSX, CTA, HLTF, ITGA10, PLCG1, TTC3, HER2, MUC-1, CEA, hTERT, STn, WT1 and / or p53.

[0254] III. Composition

[0255] In one aspect, the present disclosure further encompasses compositions comprising anti-MYCT1 antibodies, fragments thereof, or polynucleotides encoding the antibodies. In some aspects, the compositions of the present disclosure include compositions that regulate MY CT1. Specifically, the compositions that regulate MYCT1 can be compositions that downregulate MYCT1 expression and / or activity. In certain aspects, the disclosed compositions include one or more antibodies, fragments thereof, or polynucleotides encoding antibodies disclosed herein.

[0256] The composition of the present invention may further comprise a pharmaceutically acceptable excipient, carrier or diluent. In addition, the composition of the present invention may contain a preservative, a solubilizing agent, a stabilizer, a wetting agent, an emulsifier, a sweetener, a coloring agent, a flavoring agent, a salt (the substance of the present invention itself can be provided in the form of a pharmaceutically acceptable salt), a buffer, a coating agent or an antioxidant.

[0257] In some aspects, the compositions provided herein are pharmaceutical compositions comprising one or more antibodies, fragments thereof, or polynucleotides encoding antibodies disclosed herein. In some aspects, the pharmaceutical compositions herein may comprise one or more antibodies, fragments thereof, or polynucleotides encoding antibodies disclosed herein and at least one pharmaceutically acceptable carrier and / or excipient. Pharmaceutically acceptable carriers or excipients suitable for compositions described herein are well known to those skilled in the art for preserving antibodies or antibody fragments and delivering them to any mammalian subject, including humans and other mammals.

[0258] Compositions comprising anti-MYCT1 antibodies disclosed herein or fragments thereof can be formulated for administration in any convenient manner, including administration by injection, infusion or implantation of compositions. Compositions described herein can be formulated for subcutaneous, intradermal, intratumoral, intranodal, intramedullary, intramuscular, intravenous (iv) or intraperitoneal injection or administration. In some aspects, the disclosed compositions are formulated for administration by intradermal or subcutaneous injection. In some aspects, the disclosed compositions are formulated for administration by intravenous injection. Compositions can also be formulated for direct injection into tumors, lymph nodes or infection sites. In some aspects, compositions disclosed herein are formulated for intravenous administration. In some aspects, compositions disclosed herein are formulated for intraperitoneal injection.

[0259] In some respects, compositions described herein can have liposome, and the liposome contains antibody, its fragment or the polynucleotide encoding the antibody. In some respects, the liposome used herein can be generated by the lipid composition with the phosphatidylethanolamine (PEG-PE) derived from phosphatidylcholine, cholesterol and PEG by reverse phase evaporation method. In some respects, the liposome used for this paper can be extruded through the filter of limited aperture to produce the liposome with desired diameter.

[0260] In some aspects, the antibodies, fragments thereof or encoding polynucleotides herein can be encapsulated in microcapsules, such as those prepared by coacervation techniques or by interfacial polymerization, such as hydroxymethylcellulose or gelatin-microcapsules and poly-(methyl acrylate) microcapsules, respectively, in colloidal drug delivery systems (e.g., liposomes, albumin microspheres, microemulsions, nanoparticles and nanocapsules) or in macroemulsions.

[0261] In some aspects, the compositions described herein can be formulated into sustained release forms. In some aspects, the compositions to be used for in vivo administration herein can be sterile. In some aspects, this can be easily accomplished by, for example, filtration with a sterile filtration membrane. In some aspects, the therapeutic antibody compositions herein can be placed in a container with a sterile access port, such as an intravenous solution bag or a vial with a stopper that can be pierced by a hypodermic needle.

[0262] In certain aspects, the compositions described herein may be in unit dosage form, such as tablets, pills, capsules, powders, granules, solutions or suspensions, or suppositories, for oral, parenteral or rectal administration, or administration by inhalation or insufflation.

[0263] In some aspects, the emulsion compositions herein may be those prepared by mixing an antibody with Intralipid™ or its components (soybean oil, lecithin, glycerol and water).

[0264] Suitable emulsions can be prepared using commercially available fat emulsions, such as IntralipidTM, LiposynTM, InfonutrolTM, LipofundinTM and LipiphysanTM. The active ingredient can be dissolved in a premixed emulsion composition, or alternatively it can be dissolved in an oil (e.g., soybean oil, safflower oil, cottonseed oil, sesame oil, corn oil or almond oil) and form an emulsion when mixed with a phospholipid (e.g., lecithin, soybean lecithin or soybean lecithin) and water. It should be understood that other ingredients, such as glycerol or glucose, can be added to adjust the tension of the emulsion. Suitable emulsions typically contain up to about 20% oil, such as about 5% to about 20% oil. Fat emulsions can contain fat droplets of about 0.1 μm to about 1.0 μm, particularly fat droplets of about 0.1 μm to 0.5 μm, and have a pH in the range of about 5.5 to about 8.0.

[0265] In some aspects, pharmaceutical compositions for inhalation or intranasal administration herein may include solutions and suspensions in pharmaceutically acceptable aqueous or organic solvents or mixtures thereof, as well as powders. In some aspects, liquid or solid compositions herein may contain suitable pharmaceutically acceptable excipients as indicated above. In other aspects, the compositions may be administered via the oral or nasal respiratory route for local or systemic effect.

[0266] In some aspects, the composition can be in a sterile pharmaceutically acceptable solvent, can be atomized by using a gas. In some aspects, the atomized solution herein can be directly breathed from the atomizing device or the atomizing device can be attached to a mask, an oxygen tent or an intermittent positive pressure breathing machine. In some embodiments, the solution, suspension or powder composition herein can be used from a device that delivers the preparation in an appropriate manner, preferably orally or nasally.

[0267] Components of the composition

[0268] In certain aspects, the compositions disclosed herein may be pharmaceutical compositions. In this regard, the composition comprises, as an active ingredient, an anti-MYCT1 antibody as described above and at least one pharmaceutically acceptable excipient.

[0269] Pharmaceutically acceptable excipients can be diluents, binders, fillers, buffers, pH adjusters, disintegrants, dispersants, preservatives, lubricants, taste masking agents, flavoring agents or coloring agents. The amount and type of excipients used to form the pharmaceutical composition can be selected according to principles known in pharmaceutical science.

[0270] (i) Diluent

[0271] In one aspect, excipient can be diluent.Diluent can be compressible (i.e. plastically deformable) or grinding brittle.The limiting examples of suitable compressible diluent include microcrystalline cellulose (MCC), cellulose derivatives, cellulose powder, cellulose ester (i.e. acetate and butyrate mixed ester), ethylcellulose, methylcellulose, hydroxypropyl cellulose, hydroxypropyl methylcellulose, sodium carboxymethylcellulose, corn starch, phosphorylated corn starch, pregelatinized corn starch, rice starch, potato starch, tapioca starch, starch-lactose, starch-calcium carbonate, sodium starch glycolate, glucose, fructose, lactose, lactose monohydrate, sucrose, xylose, lactitol, mannitol, maltitol, sorbitol, xylitol, maltodextrin and trehalose.The limiting examples of suitable grinding brittle diluent include calcium hydrogen phosphate (anhydrous or dihydrate), calcium phosphate, calcium carbonate and magnesium carbonate.

[0272] (ii) Adhesive

[0273] In another aspect, the excipient can be a binder. Suitable binders include, but are not limited to, starch, pregelatinized starch, gelatin, polyvinyl pyrrolidone, cellulose, methyl cellulose, sodium carboxymethyl cellulose, ethyl cellulose, polyacrylamide, polyvinyloxoazolidone, polyvinyl alcohol, C 12 -C 18 Fatty acid alcohols, polyethylene glycols, polyols, sugars, oligosaccharides, polypeptides, oligopeptides and combinations thereof.

[0274] (iii) Fillers

[0275] In another aspect, the excipient can be a filler. Suitable fillers include, but are not limited to, carbohydrates, inorganic compounds, and polyvinylpyrrolidone. By way of non-limiting example, the filler can be dibasic and tribasic calcium sulfate, starch, calcium carbonate, magnesium carbonate, microcrystalline cellulose, calcium hydrogen phosphate, magnesium carbonate, magnesium oxide, calcium silicate, talc, modified starch, lactose, sucrose, mannitol, or sorbitol.

[0276] (iv) Buffer

[0277] In yet another aspect, the excipient can be a buffer. Representative examples of suitable buffers include, but are not limited to, phosphates, carbonates, citrates, tris buffers, and buffered saline salts (eg, Tris buffered saline or phosphate buffered saline).

[0278] (v) pH adjuster

[0279] In various aspects, the excipient can be a pH adjuster. By way of non-limiting example, the pH adjuster can be sodium carbonate, sodium bicarbonate, sodium citrate, citric acid or phosphoric acid.

[0280] (vi) Disintegrants

[0281] In other aspects, the excipient can be a disintegrant. Disintegrants can be non-effervescent or effervescent. Suitable examples of non-effervescent disintegrants include, but are not limited to, starches such as corn starch, potato starch, its pregelatinized and modified starches, sweeteners, clays such as bentonite, microcrystalline cellulose, alginate, sodium starch glycolate, gums such as agar, guar gum, locust bean gum, karaya gum, pectin and tragacanth. Non-limiting examples of suitable effervescent disintegrants include a combination of sodium bicarbonate and citric acid and a combination of sodium bicarbonate and tartaric acid.

[0282] (vii) Dispersants

[0283] In yet another aspect, the excipient may be a dispersant or a dispersion enhancer. Suitable dispersants may include, but are not limited to, starch, alginic acid, polyvinyl pyrrolidone, guar gum, kaolin, bentonite, purified lignocellulose, sodium starch glycolate, isoamorphous silicate, and microcrystalline cellulose.

[0284] (viii) Excipients

[0285] In another alternative aspect, the excipient can be a preservative. Non-limiting examples of suitable preservatives include antioxidants such as BHA, BHT, vitamin A, vitamin C, vitamin E or retinyl palmitate, citric acid, sodium citrate; chelating agents such as EDTA or EGTA; and antimicrobials such as parabens, chlorobutanol or phenol.

[0286] (ix) Lubricants

[0287] In a further aspect, the excipient can be a lubricant. Non-limiting examples of suitable lubricants include minerals, such as talc or silicon dioxide; and fats, such as vegetable stearin, magnesium stearate or stearic acid.

[0288] (x) Odor-masking agents

[0289] In yet another aspect, the excipient can be a taste masking agent. Taste masking materials include cellulose ethers; polyethylene glycol; polyvinyl alcohol; copolymers of polyvinyl alcohol and polyethylene glycol; monoglycerides or triglycerides; acrylic acid polymers; mixtures of acrylic acid polymers and cellulose ethers; cellulose acetate phthalate; and combinations thereof.

[0290] (xi) Flavoring agents

[0291] In an alternative aspect, the excipient may be a flavoring agent. The flavoring agent may be selected from synthetic flavoring oils and flavoring aromatics and / or natural oils, extracts from plants, leaves, flowers, fruits, and combinations thereof.

[0292] (xii) Colorants

[0293] In yet a further aspect, the excipient can be a colorant. Suitable color additives include, but are not limited to, food, drug, and cosmetic colors (FD&C), drug and cosmetic colors (D&C), or external drug and cosmetic colors (Ext. D&C).

[0294] The weight fraction of the excipient or combination of excipients in the composition can be about 99% or less, about 97% or less, about 95% or less, about 90% or less, about 85% or less, about 80% or less, about 75% or less, about 70% or less, about 65% or less, about 60% or less, about 55% or less, about 50% or less, about 45% or less, about 40% or less, about 35% or less, about 30% or less, about 25% or less, about 20% or less, about 15% or less, about 10% or less, about 5% or less, about 2% or about 1% or less of the total weight of the composition.

[0295] The present disclosure encompasses pharmaceutical compositions comprising anti-MYCT1 antibodies, fragments thereof, or polynucleotides encoding the antibodies described herein, so as to facilitate administration and promote the stability of the active agent. For example, anti-MYCT1 antibodies, fragments thereof, or polynucleotides encoding the antibodies disclosed herein can be mixed with at least one pharmaceutically acceptable carrier or excipient to produce a pharmaceutical composition that can and effectively be administered (administered) to a living subject, such as a suitable subject (i.e., a "subject in need of treatment" or a "subject in need thereof"). Methods for preparing anti-MYCT1 antibodies, fragments thereof, or polynucleotides encoding the antibodies disclosed herein and administering them to subjects in need are well known to those skilled in the art or can be easily determined by those skilled in the art. The route of administration of anti-MYCT1 antibodies, fragments thereof, or polynucleotides encoding the antibodies can be, for example, peripheral, oral, parenteral, by inhalation or topical.

[0296] Pharmaceutical compositions for effective administration are deliberately designed to be suitable for the selected mode of administration and appropriately use pharmaceutically acceptable excipients such as compatible carriers, dispersants, buffers, surfactants, preservatives, solubilizers, isotonic agents, stabilizers, and the like.

[0297] Non-limiting examples of pharmaceutically acceptable carriers include physiological saline, ion exchangers, alumina, aluminum stearate, lecithin, serum proteins such as human serum albumin, buffer substances such as phosphates, glycine, sorbic acid, potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids, water, salts or electrolytes such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silicon dioxide, magnesium trisilicate, polyvinyl pyrrolidone, cellulose-based substances, polyethylene glycol, sodium carboxymethyl cellulose, polyacrylates, waxes, polyethylene-polyoxypropylene-block polymers, polyethylene glycol, lanolin, or combinations thereof.

[0298] The prevention of the action of microorganisms can be achieved by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, ascorbic acid, thimerosal, and the like. In many cases, isotonic agents, for example, sugars, polyalcohols such as mannitol, sorbitol, or sodium chloride will be included in the composition.

[0299] Prolonged absorption of the injectable compositions can be brought about by including in the composition an agent that delays absorption, for example, aluminum monostearate and gelatin.

[0300] The compositions disclosed herein may be frozen or lyophilized for storage and reconstituted in a suitable sterile vehicle prior to use.

[0301] In some aspects, anti-MYCT1 antibodies can be formulated for parenteral administration. Preparations for parenteral administration include sterile aqueous or non-aqueous solutions, suspensions and emulsions. Examples of non-aqueous solvents are propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable organic esters such as ethyl oleate. Aqueous carriers include water, alcohol / aqueous solutions, emulsions or suspensions, including saline and buffered media. Parenteral vehicles include sodium chloride solution, Ringer's dextrose, dextrose and sodium chloride, lactated Ringer's solution, or fixed oils. Intravenous vehicles include fluid and nutritional supplements, electrolyte supplements (such as those based on Ringer's dextrose), etc. Preservatives and other additives, such as antimicrobials, antioxidants, chelating agents, and inert gases, etc., may also be present. Parenteral preparations can be single bolus doses, infusions, or loading bolus doses, followed by maintenance doses. These compositions may be administered at specific fixed or variable intervals, such as once daily or on an "as needed" basis.

[0302] Certain pharmaceutical compositions as disclosed herein can be administered orally in acceptable dosage forms, including, for example, capsules, tablets, aqueous suspensions or solutions. Certain pharmaceutical compositions can also be administered by nasal aerosol or inhalation. Such compositions can be prepared as solutions in saline, using benzyl alcohol or other suitable preservatives, absorption enhancers to enhance bioavailability, and / or other conventional solubilizing agents or dispersants.

[0303] The amount of anti-MYCT1 antibody to be combined with the carrier material to produce a single dosage form will vary depending on the host being treated and the specific mode of administration. The composition can be administered as a single dose, multiple doses, or infusion over an established period of time. The dosage regimen can also be adjusted to provide the optimal desired response (e.g., a therapeutic or preventive response).

[0304] In some respects, the concentration of antibody disclosed herein can be a predetermined concentration or standard concentration. In some respects, the concentration of antibody disclosed herein can be about 1 microgram / milliliter (mg / mL) to about 500mg / ml, about 1mg / ml to about 250mg / ml, about 1mg / ml to about 200mg / ml, about 1mg / ml to about 150mg / ml, about 1mg / ml to about 100mg / ml, about 1mg / ml to about 75mg / ml, about 1mg / ml to about 50mg / ml, about 0.1mg / ml to about 100mg / ml or other suitable concentration. In some embodiments, it is about 30mg / ml that antibody is formulated into concentration. In some embodiments, by antibody lyophilization. In some respects, before using, antibody is diluted to suitable concentration (for example in therapeutic application described below) in suitable solution.

[0305] In some aspects, in addition to the antibodies, fragments thereof or polynucleotides provided herein, the compositions described herein may further comprise an active agent. Non-limiting examples of additional active agents include, but are not limited to, antibiotics, antipyretics, antimicrobials, antifungals, NSAIDs, chemotherapeutics and anticancer agents.

[0306] In other aspects, a composition comprising a disclosed anti-MYCT1 antibody, a fragment thereof, or a polynucleotide encoding the antibody may be administered to a subject in need. In some aspects, the disclosed composition may be administered to a subject in need, wherein the subject is a human subject. In some aspects, the disclosed composition may be administered to a subject in need, wherein the subject is a subject suffering from angiogenesis-related disease, suspected of suffering from angiogenesis-related disease, or at risk of angiogenesis-related disease. In some aspects, angiogenesis-related diseases are age-related macular degeneration, ischemic retinopathy, intraocular neovascularization, corneal neovascularization, retinal neovascularization, choroidal neovascularization, diabetic macular edema, diabetic retinal ischemia, diabetic retinal edema, diabetic retinopathy, cancer, rheumatoid arthritis, endometriosis, or alopecia.

[0307] In some aspects, the disclosed compositions can be administered to a subject in need, wherein the subject suffers from cancer, is suspected of having cancer, or is at risk of cancer. In some aspects, cancer is a cancer comprising endogenous MYCT1 expression or activity of cancer cells with an increase. When compared with cancer cells between patient samples, such cancer cells express MYCT1 at a level higher than normal cells (e.g., control breast cells) or cancer cells. Such cells express MYCT1 at a level higher than the average level of expression of a colony of cancer cells (e.g., the average level of MYCT 1 expression in breast cancer cells). In some aspects, when compared with cancer cells between patient samples, cancer cells express MYCT1 at the following levels: at least 10% higher than normal cells (e.g., control breast cells) or at least 10% higher than cancer cells, for example, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70% or higher.

[0308] In some aspects, cancer is a sarcoma. In some aspects, cancer is Ewing's sarcoma, soft tissue sarcoma, liposarcoma, epithelioid sarcoma, malignant peripheral nerve sheath tumor, synovial sarcoma, Kaposi's sarcoma, osteosarcoma, fibrosarcoma, cutaneous fibrosarcoma, aggressive fibromatosis, rhabdomyosarcoma, leiomyosarcoma, undifferentiated pleomorphic sarcoma, chondrosarcoma, gastrointestinal stromal tumor or uterine sarcoma. In some aspects, cancer is breast cancer. In some aspects, cancer is ductal carcinoma in situ, invasive ductal carcinoma, inflammatory breast cancer or metastatic breast cancer.

[0309] In some aspects, the composition comprising the disclosed anti-MYCT1 antibody, its fragment or the polynucleotide encoding the antibody can reduce and / or inhibit the activity, function, expression and / or accumulation of MYCT1. In some aspects, the composition comprising the disclosed anti-MYCT1 antibody, its fragment or the polynucleotide encoding the antibody causes the activity, function, expression and / or accumulation of MYCT1 to be reduced in the subject. In some aspects, compared with the activity, function, expression and / or accumulation of MYCT1 before (baseline) or in a control subject who has not received treatment, the composition comprising the antibody, its fragment or the polynucleotide encoding the anti-MYCT1 described herein causes the activity, function, expression and / or accumulation of MYCT1 to be reduced by at least about 10%, for example, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95% or more.

[0310] In some aspects, the composition comprising an anti-MYCT1 antibody, a fragment thereof, or a polynucleotide encoding the antibody reduces tumor volume in a subject by at least 10%, e.g., 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95% or more, compared to the tumor volume before administration of the anti-MYCT1 antibody, fragment thereof, or polynucleotide encoding the antibody (baseline) or in an untreated control subject.

[0311] In some aspects, compared to the tumor volume of a control subject before (baseline) or not receiving treatment, the composition comprising an anti-MYCT1 antibody, a fragment thereof, or a polynucleotide encoding the antibody reduces the tumor volume in the subject by at least 10%, such as 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95% or more. The tumor volume of a subject can be assessed using any conventional method known in the art, non-limiting examples of which include diagnostic techniques such as X-rays, CT scans, bone marrow biopsies, serum protein electrophoresis (SPEP) or MRI.

[0312] In some aspects, compared to the tumor angiogenesis of a control subject before (baseline) or not receiving treatment, the composition comprising an anti-MYCT1 antibody, a fragment thereof, or a polynucleotide encoding the antibody reduces tumor angiogenesis in the subject by at least 10%, such as 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95% or more. Tumor angiogenesis in a subject can be assessed using any conventional method known in the art, non-limiting examples of which include diagnostic techniques, such as counting the number of blood vessels in a given area using a stereomicroscope or by measuring the expression of biomarkers associated with angiogenesis, including α v β3 integrin, endoglin, and vascular endothelial growth factor (VEGF) receptor (VEGFR) 2.

[0313] In some aspects, compared to the immune response or anti-tumor immunity of a control subject before (baseline) or untreated, the composition comprising an anti-MYCT1 antibody, a fragment thereof, or a polynucleotide encoding the antibody can enhance the immune response or anti-tumor immunity of the subject against cancer cells by at least 10%, for example, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or more. The immune response against cancer cells can be measured using biomarkers reflecting the tumor immune microenvironment and the intrinsic characteristics of tumor cells, such as PD-L1 expression and the density of tumor infiltrating lymphocytes (TILs).

[0314] In other aspects, the compositions of the present disclosure may optionally include one or more immune checkpoint inhibitor compounds. Inhibitors can target any immune checkpoint known in the art, including but not limited to CTLA-4, PDL1, PDL2, PD1, B7-H3, B7-H4, BTLA, HVEM, TIM3, GAL9, LAG 3, CSF-1R, VISTA, KIR, 2B4, CD160, CGEN-15049, CHK1, CHK2, A2aR, CD28, CD86, CD69, CD48, CD113, CEACAM-1, galectin-1, TIGIT, GPR56, CD48, GARP, PD1H, LAIR1, TIM1, TIM4 and B-7 family ligands. In some aspects, immune checkpoint inhibitors include CTLA-4 blocking antibodies (ipilimumab (Yervoy), tremelimumab (Imjuno)), PD-1 inhibitors (pembrolizumab (Keytruda), nivolumab (Opdivo), cimipril (Libt ayo), CT-011 (pidilizumab), AMP224), PD-L1 inhibitors (atezolizumab (tecentriq), avelumab (Bavencio), durvalumab (Imfinzi), BMS-936559), Lag3 inhibitors (relaximab), combinations of Lag3 and PD1 inhibitors (PD-1 inhibitor nivolumab (Opdualag), OX40 inhibitors (MEDI6469), CD160 inhibitors (BY55). Non-limiting examples of inhibitors of CSF-1R include PLX3397, PLX486, RG7155, AMG820, ARRY-382, FPA008, IMC-CS4, JNJ-40346527, MCS110, or any combination thereof.

[0315] In certain aspects, the compositions of the present disclosure may include one or more PD-1 inhibitors and / or PD-L1 inhibitor compounds. Non-limiting examples of immune checkpoint compounds include monoclonal antibodies targeting PD-1 or PD-L1. In some aspects, monoclonal antibodies targeting PD-1 or PD-L1 include pembrolizumab, nivolumab, cemipril, atezolizumab, avelumab, durvalumab and / or ipilimumab.

[0316] In some aspects, an anti-MYCT1 antibody, a fragment thereof, or a polynucleotide encoding the antibody in combination with an immune checkpoint inhibitor compound can be administered to a subject with a cancer that was initially sensitive to immune checkpoint inhibitor treatment and subsequently became insensitive to immune checkpoint inhibitor treatment. In some aspects, an anti-MYCT1 antibody, a fragment thereof, or a polynucleotide encoding the antibody in combination with an immune checkpoint inhibitor compound can be administered to a subject with a cancer that is insensitive to immune checkpoint inhibitor treatment. In some aspects, the disclosed anti-MYCT1 antibodies, fragments thereof, or polynucleotides encoding the antibodies sensitize cancer to immune checkpoint inhibitors.

[0317] In some aspects, compared to the tumor volume of a control subject who has not received treatment or a subject who has received an antibody, a fragment thereof, or a polynucleotide encoding the antibody or a subject who has received an immune checkpoint inhibitor compound alone (baseline), an anti-MYCT1 antibody, a fragment thereof, or a polynucleotide encoding the antibody and an immune checkpoint inhibitor compound before administering the antibody, a fragment thereof, or a polynucleotide encoding the antibody and an immune checkpoint inhibitor compound, the combination can reduce the tumor volume of the subject by at least 10%, for example, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or more. In some aspects, the combination of anti-MYCT1 antibodies, fragments thereof, or polynucleotides encoding the antibodies and immune checkpoint inhibitor compounds reduces the tumor volume of the subject by at least 25%. Any conventional method known in the art can be used to assess the tumor volume of the subject, and non-limiting examples of the conventional method include diagnostic techniques, such as X-rays, CT scans, bone marrow biopsies, serum protein electrophoresis (SPEP) or MRI.

[0318] In some aspects, the combination of anti-MYCT1 antibodies, fragments thereof, or polynucleotides encoding the antibodies and immune checkpoint inhibitor compounds reduces tumor angiogenesis in the subject by at least 10%, such as 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95% or more, compared to tumor angiogenesis in untreated control subjects or subjects receiving antibodies, fragments thereof, or polynucleotides encoding the antibodies alone or subjects receiving immune checkpoint inhibitor compounds alone before administration of anti-MYCT1 antibodies, fragments thereof, or polynucleotides encoding the antibodies (baseline). Tumor angiogenesis in a subject can be assessed using any conventional method known in the art, non-limiting examples of which include diagnostic techniques, such as counting the number of blood vessels in a given area using a stereomicroscope or by measuring the expression of biomarkers associated with angiogenesis, including α v β3 integrin, endoglin, and vascular endothelial growth factor (VEGF) receptor (VEGFR) 2.

[0319] In some aspects, the combination of anti-MYCT1 antibodies, fragments thereof, or polynucleotides encoding the antibodies and immune checkpoint inhibitor compounds can enhance the immune response or anti-tumor immunity of the subject against cancer cells by at least 10%, for example, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or more, compared to the immune response or anti-tumor immunity of a control subject who has not received treatment or a subject who has received an antibody, a fragment thereof, or a polynucleotide encoding the antibody or a subject who has received an immune checkpoint inhibitor compound alone before (baseline) administration of the antibody, a fragment thereof, or a polynucleotide encoding the antibody and an immune checkpoint inhibitor compound. The combination of anti-MYCT1 antibodies, fragments thereof, or polynucleotides encoding the antibodies and immune checkpoint inhibitor compounds can enhance the immune response or anti-tumor immunity of the subject against cancer cells by at least 10%, for example, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or more. The immune response against cancer cells can be measured using biomarkers that reflect the intrinsic characteristics of the tumor immune microenvironment and tumor cells, such as PD-L1 expression and the density of tumor infiltrating lymphocytes (TILs).

[0320] Further, the compositions of the present disclosure may optionally include one or more VEGF inhibitors. In non-limiting examples, the compositions of the present disclosure may optionally include one or more of the following: axitinib, bevacizumab, cabozantinib, lapatinib, lenvatinib, pazopanib, ponatinib, ramucirumab, ranibizumab, regorafenib, sorafenib, sunitinib and / or vandetanib.

[0321] In some aspects, the combination of anti-MYCT1 antibodies, fragments thereof, or polynucleotides encoding the antibodies and VEGF inhibitors can reduce the tumor volume of the subject by at least 10%, such as 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or more, compared to the tumor volume of untreated control subjects or subjects receiving antibodies, fragments thereof, or polynucleotides encoding the antibodies or VEGF inhibitors alone before administration of the antibody, fragments thereof, or polynucleotides encoding the antibodies and VEGF inhibitors (baseline). In some aspects, the combination of anti-MYCT1 antibodies, fragments thereof, or polynucleotides encoding the antibodies and VEGF inhibitor compounds reduces the tumor volume of the subject by at least 25%. The tumor volume of the subject can be assessed using any conventional method known in the art, non-limiting examples of which include diagnostic techniques, such as X-rays, CT scans, bone marrow biopsies, serum protein electrophoresis (SPEP) or MRI.

[0322] In some aspects, the combination of anti-MYCT1 antibodies, fragments thereof, or polynucleotides encoding the antibodies and VEGF inhibitors reduces tumor angiogenesis in a subject by at least 10%, such as 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95% or more, compared to tumor angiogenesis in untreated control subjects or subjects receiving antibodies, fragments thereof, or polynucleotides encoding the antibodies alone or receiving VEGF inhibitors alone before administration of anti-MYCT1 antibodies, fragments thereof, or polynucleotides encoding the antibodies (baseline). Tumor angiogenesis in a subject can be assessed using any conventional method known in the art, non-limiting examples of which include diagnostic techniques, such as counting the number of blood vessels in a given area using a stereomicroscope or by measuring the expression of biomarkers associated with angiogenesis, including α v β3 integrin, endoglin, and vascular endothelial growth factor (VEGF) receptor (VEGFR) 2.

[0323] In some aspects, the combination of anti-MYCT1 antibodies, fragments thereof, or polynucleotides encoding the antibodies and VEGF inhibitor compounds can enhance the immune response or anti-tumor immunity of the subject against cancer cells by at least 10%, for example, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or more, compared to the immune response or anti-tumor immunity of untreated control subjects or subjects receiving antibodies, fragments thereof, or polynucleotides encoding the antibodies or subjects receiving VEGF inhibitors alone before administration of the antibody, fragments thereof, or polynucleotides encoding the antibodies and VEGF inhibitors (baseline). The immune response against cancer cells can be measured using biomarkers that reflect the tumor immune microenvironment and the intrinsic characteristics of tumor cells, such as PD-L1 expression and the density of tumor infiltrating lymphocytes (TILs).

[0324] As noted above, the agents or compositions described herein may also be used in combination with other therapeutic agents, as further described below. Therefore, in addition to the therapies described herein, other therapies known to be effective for treating diseases, disorders or conditions may also be provided to the subject. In various examples, the method further includes administering to the patient additional cancer treatments. In some examples, additional cancer treatments are selected from surgery, radiotherapy, chemotherapy, toxin therapy, immunotherapy, cryotherapy, gene therapy and combinations thereof. Examples of useful anti-angiogenic therapeutic targets according to the present disclosure include EGF, VEGF, FGF and matrix remodeling proteins. In various examples, chemotherapeutic agents are drugs or pharmaceutical preparations. Non-limiting examples of pharmaceutical preparations include polymer micelle preparations, liposome preparations, dendrimer preparations, polymer-based nanoparticle preparations, silica-based nanoparticle preparations, nanoscale coordination polymer preparations, nanoscale metal-organic framework preparations, inorganic nanoparticle preparations, etc.

[0325] Various chemotherapeutic agents (e.g., chemotherapeutic drugs) can be used. Any FDA-approved chemotherapeutic agent (e.g., chemotherapeutic drug) can be used. Combinations of chemotherapeutic agents can be used. Chemotherapeutic agents and anticancer agents commonly used to treat cancer and that can be used in combination with any of the compositions disclosed herein include, but are not limited to, platinum compounds (e.g., cisplatin or carboplatin) and taxane compounds, such as paclitaxel. Docetaxel Other chemotherapeutic agents used to treat cancer and that can be used in combination with any of the compositions disclosed herein include, but are not limited to, nab-paclitaxel (nab-paclitaxel, ), Pemetrexed Irinotecan (CPT-11, ), cyclophosphamide Liposomal doxorubicin Gemcitabine Hexamethylmelamine Ifosfamide Melphalan (Alkeran), Vinorelbine Topotecan (Hycamtin), etoposide (VP-16), and capecitabine

[0326] In some aspects, the additional drug or therapeutic agent can be a genotoxic agent (e.g., a DNA damaging agent or drug). As used herein, "genotoxic therapy" refers to the treatment of a tumor or cancer that utilizes the destructive properties of the treatment to induce DNA damage to the tumor or cancer cell. The treatment is traditionally part of a standardized regimen. As proliferation continues, any damage to the tumor cancer will be passed on to the next generation of cancer cells. If this damage is severe enough, it will induce the cell to undergo apoptosis. In non-limiting examples, genotoxic therapy can include gamma-irradiation, alkylating agents such as nitrogen mustards (chlorambucil, cyclophosphamide, ifosfamide, melphalan), nitrosoureas (streptozocin, carmustine, lomustine), alkyl sulfonates (busulfan), triazines (dacarbazine, temozolomide) and ethyleneimines (thiotepa, hexamethylmelamine), platinum drugs such as cisplatin, carboplatin, oxalaplatin, antimetabolites such as 5-fluorouracil, 6-mercaptopurine, capecitabine, cladribine, clofarabine, cytarabine, floxuridine, fludarabine, gemcitabine, hydroxyurea, methotrexate , pemetrexed, pentostatin, thioguanine, anthracyclines such as daunorubicin, doxorubicin, epirubicin, idarubicin, antitumor antibiotics such as dactinomycin-D, bleomycin, mitomycin-C, mitoxantrone, topoisomerase inhibitors such as topoisomerase I inhibitors (topotecan, irinotecan) and topoisomerase II inhibitors (etoposide, teniposide, mitoxantrone), mitotic inhibitors such as taxanes (paclitaxel, docetaxel), epothilones (ixabepilone), vinca alkaloids (vinblastine, vincristine, vinorelbine) and estramustine.

[0327] Treatment according to the methods described herein can be performed prior to, concurrently with, or after conventional treatments for cancer or tumors.

[0328] The dosage of the additional drug or therapeutic agent can vary between wide limits, depending on the disease or condition to be treated, the age and condition of the subject to be treated. In aspects where a composition further comprising at least one additional drug or therapeutic agent is contacted with the sample, the concentration of the at least one additional drug or therapeutic agent can be about 0.01 μM to about 10 μM. Alternatively, the concentration of the at least one additional drug or therapeutic agent can be about 0.01 μM to about 5 μM. For example, the concentration of the at least one additional drug or therapeutic agent can be about 0.01 μM, about 0.05 μM, about 0.1 μM, about 0.2 μM, about 0.3 μM, about 0.4 μM, about 0.5 μM, about 0.6 μM, about 0.7 μM, about 0.8 μM, about 0.9 μM, about 1 μM, about 2 μM, about 3 μM, about 4 μM, about 5 μM, about 6 μM, about 7 μM, about 8 μM, about 9 μM or about 10 μM. In addition, the concentration of the at least one additional drug or therapeutic agent is greater than 10 μM. For example, the concentration of the at least one additional agent can be about 10 μM, about 15 μM, about 20 μM, about 25 μM, about 30 μM, about 35 μM, about 40 μM, about 45 μM, about 50 μM, about 55 μM, about 60 μM, about 65 μM, about 70 μM, about 75 μM, about 80 μM, about 85 μM, about 90 μM, about 95 μM, or about 100 μM.

[0329] In the aspect that the composition further comprises at least one other drug or therapeutic agent administered to the subject, the dosage of the other drug or therapeutic agent can be about 0.1mg / kg to about 500mg / kg. For example, the dosage of at least one other drug or therapeutic agent can be about 0.1mg / kg, about 0.5mg / kg, about 1mg / kg, about 5mg / kg, about 10mg / kg, about 15mg / kg, about 20mg / kg or about 25mg / kg. Alternatively, the dosage of the at least one other drug or therapeutic agent can be about 25mg / kg, about 50mg / kg, about 75mg / kg, about 100mg / kg, about 125mg / kg, about 150mg / kg, about 175mg / kg, about 200mg / kg, about 225mg / kg or about 250mg / kg. Additionally, the dosage of the at least one additional drug or therapeutic agent may be about 300 mg / kg, about 325 mg / kg, about 350 mg / kg, about 375 mg / kg, about 400 mg / kg, about 425 mg / kg, about 450 mg / kg, about 475 mg / kg, or about 500 mg / kg.

[0330] In general, a safe and effective amount of the composition is administered, such as an amount that causes a desired therapeutic effect in a subject while minimizing undesirable side effects. In various aspects, an effective amount of the composition described herein can substantially reduce the growth or spread of a subject's cancer. In some aspects, an effective amount is an amount that can treat a cancer or tumor. In some aspects, an effective amount is an amount that can treat one or more symptoms associated with a cancer or tumor.

[0331] The amount of the compositions described herein that can be combined with a pharmaceutically acceptable carrier to produce a single dosage form will vary depending on the host being treated and the particular mode of administration. It will be appreciated by those skilled in the art that the unit content of the medicament contained in the individual doses of each dosage form need not constitute a therapeutically effective amount in itself, since the necessary therapeutically effective amount can be achieved by administering multiple individual doses.

[0332] The toxicity and therapeutic efficacy of the compositions described herein can be determined by measuring the LD 50 (a dose lethal to 50% of the population) and ED 50 The dose ratio between toxic and therapeutic effects is the therapeutic index, which can be expressed as LD 50 / ED 50 The larger the therapeutic index, the better.

[0333] The specific therapeutically effective dosage level for any particular subject will depend on a variety of factors, including the condition being treated and the severity of the condition; the activity of the specific compound employed; the specific composition employed; the age, weight, general health, sex, and diet of the subject; the time of administration; the route of administration; the rate of excretion of the composition employed; the duration of the treatment; drugs used in combination or concomitantly with the specific compound employed; and like factors well known in the medical arts (see, e.g., Koda-Kimble et al. (2004) Applied Therapeutics: The Clinical Use of Drugs, Lippincott Williams & Wilkins, ISBN 0781748453; Winter (2003) Basic Clinical Pharmacokinetics, 4th ed., Lippincott Williams & Wilkins, ISBN 0781741475; Sharqel (2004) Applied Biopharmaceutics & Pharmacokinetics, McGraw-Hill / Appleton & Lange, ISBN 0071375503). For example, it is well known to those skilled in the art to start the dosage of the composition at a level lower than that required to achieve the desired therapeutic effect and gradually increase the dosage until the desired effect is achieved. If desired, the effective daily dose can be divided into multiple doses for administration purposes. Therefore, a single dose composition can contain such an amount or its submultiple constituting the daily dose. However, it should be understood that the total daily dosage of the compounds and compositions of the present disclosure will be determined by the attending physician within the scope of reasonable medical judgment.

[0334] The administration of the compositions disclosed herein can occur as a single event or over the time course of treatment. For example, the composition can be administered daily, weekly, biweekly, or monthly. For the treatment of acute conditions, the time course of treatment is generally at least a few days. Certain conditions can extend treatment from a few days to a few weeks. For example, treatment can be extended over a week, two weeks, or three weeks. For more chronic conditions, treatment can be extended from a few weeks to a few months or even a year or longer.

[0335] In the case where there is more than one administration in the methods of the invention, the administrations may be separated by time intervals of one, two, three, four, five, six, seven, eight, nine, ten or more minutes, by intervals of about one, two, three, four, five, six, seven, eight, nine, ten, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24 hours, and so forth. In the context of hours, the term "about" means any time interval within plus or minus 30 minutes. Administrations may also be separated by time intervals of one, two, three, four, five, six, seven, eight, nine, ten, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 days, and combinations thereof. The present disclosure is not limited to dosing intervals that are equally spaced in time, but rather encompasses dosing at non-equal intervals, such as a priming schedule consisting of administration on Days 1, 4, 7, and 25, merely to provide a non-limiting example.

[0336] For example, dosing schedules of once a week, twice a week, three times a week, four times a week, five times a week, six times a week, seven times a week, once every two weeks, once every three weeks, once every four weeks, once every five weeks, etc. can be used in the present disclosure. The dosing schedule encompasses administration for a total period of time, such as one week, two weeks, three weeks, four weeks, five weeks, six weeks, two months, three months, four months, five months, six months, seven months, eight months, nine months, ten months, eleven months, and twelve months.

[0337] The above dosing schedule is provided in cycles. The cycles may be repeated about, for example, every seven days; every 14 days; every 21 days; every 28 days; every 35 days; 42 days; every 49 days; every 56 days; every 63 days; every 70 days; etc. Non-dosing intervals may occur between cycles, where the intervals may be about, for example, seven days; 14 days; 21 days; 28 days; 35 days; 42 days; 49 days; 56 days; 63 days; 70 days; etc. In this context, the term "about" means plus or minus one day, plus or minus two days, plus or minus three days, plus or minus four days, plus or minus five days, plus or minus six days, or plus or minus seven days.

[0338] IV. Methods

[0339] The present disclosure further provides a method for treating a subject with cancer. In some aspects, the methods described herein may include administering an effective amount of one or more anti-MY CT1 antibodies disclosed herein or fragments thereof or one or more pharmaceutical compositions disclosed herein to a subject in need. In some aspects, the methods herein may include administering to a subject in need, wherein the subject is a human subject. In some aspects, the methods herein may include administering to a subject in need, wherein the subject is a subject suffering from angiogenesis-related diseases, suspected of suffering from angiogenesis-related diseases, or at risk of angiogenesis-related diseases. In some aspects, angiogenesis-related diseases are age-related macular degeneration, ischemic retinopathy, intraocular neovascularization, corneal neovascularization, retinal neovascularization, choroidal neovascularization, diabetic macular edema, diabetic retinal ischemia, diabetic retinal edema, diabetic retinopathy, cancer, rheumatoid arthritis, endometriosis, or alopecia.

[0340] In some aspects, the methods herein may include administration to a subject having cancer, suspected of having cancer, or at risk of cancer. In some aspects, cancer is a cancer comprising cancer cells with increased endogenous MYCT1 expression. When compared to cancer cells between patient samples, such cancer cells express MYCT1 at a higher level than normal cells (e.g., control breast cells) or cancer cells. Such cells express MYCT1 at a higher level than the average level of cancer cell population expression (e.g., the average level of MYCT1 expression in breast cancer cells). In some aspects, when compared to cancer cells between patient samples, cancer cells express MYCT1 at a level that is at least 10% higher than normal cells (e.g., control breast cells) or at least 10% higher than cancer cells (e.g., 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70% or more). In this regard, the method includes administering to a subject a therapeutically effective amount of a plurality of one or more anti-MYCT1 antibodies disclosed herein or fragments thereof or one or more pharmaceutical compositions disclosed herein.

[0341] In some aspects of the method, the cancer is a sarcoma. In some aspects, the cancer is Ewing's sarcoma, soft tissue sarcoma, liposarcoma, epithelioid sarcoma, malignant peripheral nerve sheath tumor, synovial sarcoma, Kaposi's sarcoma, osteosarcoma, fibrosarcoma, cutaneous fibrosarcoma, aggressive fibromatosis, rhabdomyosarcoma, leiomyosarcoma, undifferentiated pleomorphic sarcoma, chondrosarcoma, gastrointestinal stromal tumor or uterine sarcoma. In some aspects, the cancer is breast cancer. In some aspects, the cancer is ductal carcinoma in situ, invasive ductal carcinoma, inflammatory breast cancer or metastatic breast cancer.

[0342] In some aspects, the administration of a disclosed anti-MYCT1 antibody, a fragment thereof, or a polynucleotide encoding the antibody can reduce and / or inhibit the activity, function, expression, and / or accumulation of MYCT1. In some aspects, the administration of a composition comprising a disclosed anti-MYCT1 antibody, a fragment thereof, or a polynucleotide encoding the antibody causes a reduction in the activity, function, expression, and / or accumulation of MYCT1 in a subject. In some aspects, the administration of an antibody, a fragment thereof, or a polynucleotide encoding the antibody before (baseline) or in a control subject who has not been treated causes the activity, function, expression, and / or accumulation of MYCT1 to be reduced by at least about 10%, for example, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, or more.

[0343] In some aspects, administration of an anti-MYCT1 antibody, a fragment thereof, or a polynucleotide encoding the antibody reduces the tumor volume of the subject by at least 10%, such as 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95% or more, compared to the tumor volume of a control subject before administration of the anti-MYCT1 antibody, a fragment thereof, or a polynucleotide encoding the antibody (baseline) or not receiving treatment. .

[0344] In some aspects, administration of an anti-MYCT1 antibody, a fragment thereof, or a polynucleotide encoding the antibody reduces the tumor volume of the subject by at least 10%, such as 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95% or more, compared to the tumor volume of a control subject before (baseline) or not receiving treatment, administered with an anti-MYCT1 antibody, a fragment thereof, or a polynucleotide encoding the antibody. Tumor volume in a subject can be assessed using any conventional method known in the art, non-limiting examples of which include diagnostic techniques such as X-rays, CT scans, bone marrow biopsies, serum protein electrophoresis (SPEP) or MRI.

[0345] In some aspects, administration of an anti-MYCT1 antibody, a fragment thereof, or a polynucleotide encoding the antibody reduces tumor angiogenesis in a subject by at least 10%, such as 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95% or more, compared to tumor angiogenesis in a control subject before (baseline) administration of an anti-MYCT1 antibody, a fragment thereof, or a polynucleotide encoding the antibody. Tumor angiogenesis in a subject can be assessed using any conventional method known in the art, non-limiting examples of which include diagnostic techniques, such as counting the number of blood vessels in a given area using a stereomicroscope or by measuring the expression of biomarkers associated with angiogenesis, including α v β3 integrin, endoglin, and vascular endothelial growth factor (VEGF) receptor (VEGFR) 2.

[0346] In some aspects, administration of an anti-MYCT1 antibody, a fragment thereof, or a polynucleotide encoding the antibody can enhance the immune response or anti-tumor immunity of a subject against cancer cells by at least 10%, for example, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or more, compared to the immune response or anti-tumor immunity of a control subject before (baseline) or in an untreated control subject administered with the antibody, its fragment, or a polynucleotide encoding the antibody. The immune response or anti-tumor immunity of the subject can be enhanced by at least 10%, for example, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or more. The immune response against cancer cells can be measured using biomarkers reflecting the tumor immune microenvironment and intrinsic characteristics of tumor cells, such as PD-L1 expression and the density of tumor infiltrating lymphocytes (TILs).

[0347] In other aspects, the methods of the present disclosure may optionally include administering one or more immune checkpoint inhibitor compounds. In some aspects, immune checkpoint inhibitor compounds may be administered before, during, or after administering a disclosed antibody, a fragment thereof, or a polynucleotide encoding the antibody. In some aspects, the immune checkpoint inhibitor can be an inhibitor of CTLA-4, PDL1, PDL2, PD1, B7-H3, B7-H4, BTLA, HVEM, TIM3, GAL9, LAG3, CSF-1R, VISTA, KIR, 2B4, CD160, CGEN-15049, CHK1, CHK2, A2aR, CD28, CD 86, CD69, CD48, CD113, CEACAM-1, galectin-1, TIGIT, GPR56, CD48, GARP, PD1H, LAIR1, TIM1, TIM4 and B-7 family ligands, or any combination thereof. In some aspects, the immune checkpoint inhibitors used in the methods herein include CTLA-4 blocking antibodies (ipilimumab (Yervoy), tremelimumab (Imjuno)), PD-1 inhibitors (pembrolizumab (Keytruda), nivolumab (Opdivo), cemipril (Libtayo), CT-011 (pidilizumab), AMP224), PD-L1 inhibitors (atezolizumab (tecentriq), avelumab (Bavencio), durvalumab (Imfinzi), BM S-936559), Lag3 inhibitors (relalizumab), combinations of Lag3 and PD1 inhibitors (PD-1 inhibitor nivolumab (Opdualag), OX40 inhibitors (MEDI6469), CD160 inhibitors (BY55). Non-limiting examples of inhibitors of CSF-1R include PLX3397, PLX486, RG7155, AMG820, ARRY-382, FPA008, IMC-CS4, JNJ-40346527, MCS110, or any combination thereof.

[0348] In certain aspects, the method may include administering one or more PD-1 inhibitors and / or PD-L1 inhibitor compounds. Non-limiting examples of PD-1 inhibitors and / or PD-L1 inhibitor compounds include monoclonal antibodies targeting PD-1 or PD-L1. In some aspects, monoclonal antibodies targeting PD-1 or PD-L1 include pembrolizumab, nivolumab, cemipril, atezolizumab, avelumab, durvalumab, and / or ipilimumab.

[0349] In another aspect, a method for treating cancer in a subject who is insensitive or resistant to immune checkpoint inhibitor monotherapy is provided. In this regard, the subject is diagnosed with cancer or suspected of having cancer or developing cancer, and the cancer is resistant or insensitive to immune checkpoint inhibitors. In this regard, the combined therapy disclosed herein has a tumor growth inhibition greater than the accumulation, which can be measured using any suitable measurement, and the non-limiting examples of the measurement include measuring tumor volume. The combined therapy can be combined with any other conventional cancer therapy, including but not limited to surgery and chemotherapy.

[0350] In some aspects of the method, an anti-MYCT1 antibody, a fragment thereof, or a polynucleotide encoding the antibody is administered in combination with an immune checkpoint inhibitor compound to a subject with cancer that was initially sensitive to immune checkpoint inhibitor treatment and subsequently became insensitive to immune checkpoint inhibitor treatment. In some aspects, an anti-MYCT1 antibody, a fragment thereof, or a polynucleotide encoding the antibody is administered in combination with an immune checkpoint inhibitor compound to a subject with a cancer that is insensitive to immune checkpoint inhibitor treatment. In some aspects, the disclosed anti-MYCT1 antibodies, fragments thereof, or polynucleotides encoding the antibodies sensitize cancer to immune checkpoint inhibitors.

[0351] In some aspects, administration of an anti-MYCT1 antibody, a fragment thereof, or a polynucleotide encoding the antibody in combination with an immune checkpoint inhibitor compound can reduce the tumor volume of the subject by at least 10%, for example, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or more, compared to the tumor volume of an untreated control subject or a subject receiving an antibody, a fragment thereof, or a polynucleotide encoding the antibody alone or a subject receiving an immune checkpoint inhibitor compound alone before administration of the antibody, a fragment thereof, or a polynucleotide encoding the antibody in combination with an immune checkpoint inhibitor compound (baseline). In some aspects, administration of an anti-MYCT1 antibody, a fragment thereof, or a polynucleotide encoding the antibody in combination with an immune checkpoint inhibitor compound reduces the tumor volume of the subject by at least 25%. Tumor volume in a subject can be assessed using any conventional method known in the art, non-limiting examples of which include diagnostic techniques such as X-ray, CT scan, bone marrow biopsy, serum protein electrophoresis (SPEP), or MRI.

[0352] In some aspects, administration of an anti-MYCT1 antibody, a fragment thereof, or a polynucleotide encoding the antibody in combination with an immune checkpoint inhibitor compound reduces tumor angiogenesis in a subject by at least 10%, such as 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95% or more, compared to tumor angiogenesis in an untreated control subject or a subject receiving an antibody, a fragment thereof, or a polynucleotide encoding the antibody alone or a subject receiving an immune checkpoint inhibitor compound alone before administration of an anti-MYCT1 antibody, a fragment thereof, or a polynucleotide encoding the antibody (baseline). Tumor angiogenesis in a subject can be assessed using any conventional method known in the art, non-limiting examples of which include diagnostic techniques, such as counting the number of blood vessels in a given area using a stereomicroscope or by measuring the expression of biomarkers associated with angiogenesis, including α v β3 integrin, endoglin, and vascular endothelial growth factor (VEGF) receptor (VEGFR) 2.

[0353] In some aspects, administration of an anti-MYCT1 antibody, a fragment thereof, or a polynucleotide encoding the antibody in combination with an immune checkpoint inhibitor compound can enhance the subject's immune response or anti-tumor immunity against cancer cells by at least 10%, for example, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or more, compared to the immune response or anti-tumor immunity of an untreated control subject or a subject receiving an antibody, a fragment thereof, or a polynucleotide encoding the antibody or a subject receiving an immune checkpoint inhibitor compound alone before (baseline) administration of the antibody, a fragment thereof, or a polynucleotide encoding the antibody in combination with an immune checkpoint inhibitor compound. Biomarkers that reflect the intrinsic characteristics of the tumor immune microenvironment and tumor cells, such as PD-L1 expression and the density of tumor infiltrating lymphocytes (TILs), can be used to measure the immune response against cancer cells.

[0354] Further, the methods of treatment described herein may optionally include administering one or more VEGF inhibitors. In some aspects, the VEGF inhibitor may be administered before administering a disclosed antibody, a fragment thereof, or a polynucleotide encoding the antibody, during administering a disclosed antibody, a fragment thereof, or a polynucleotide encoding the antibody, or after administering a disclosed antibody, a fragment thereof, or a polynucleotide encoding the antibody. In a non-limiting example, the compositions of the present disclosure may optionally include one or more of axitinib, bevacizumab, cabozantinib, lapatinib, lenvatinib, pazopanib, ponatinib, ramucirumab, ranibizumab, regorafenib, sorafenib, sunitinib, and / or vandetanib.

[0355] In some aspects, administration of an anti-MYCT1 antibody, a fragment thereof, or a polynucleotide encoding the antibody in combination with a VEGF inhibitor can reduce the tumor volume of the subject by at least 10%, for example, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or more, compared to the tumor volume of a control subject who did not receive treatment or a subject who received an antibody, a fragment thereof, or a polynucleotide encoding the antibody or a VEGF inhibitor alone before (baseline). Administration of an anti-MYCT1 antibody, a fragment thereof, or a polynucleotide encoding the antibody in combination with a VEGF inhibitor compound can reduce the tumor volume of the subject by at least 10%, for example, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or more. In some aspects, administration of an anti-MYCT1 antibody, a fragment thereof, or a polynucleotide encoding the antibody in combination with a VEGF inhibitor compound reduces the tumor volume of the subject by at least 25%. Tumor volume in a subject can be assessed using any conventional method known in the art, non-limiting examples of which include diagnostic techniques, such as X-rays, CT scans, bone marrow biopsies, serum protein electrophoresis (SPEP) or MRI.

[0356] In some aspects, administration of an anti-MYCT1 antibody, a fragment thereof, or a polynucleotide encoding the antibody in combination with a VEGF inhibitor reduces tumor angiogenesis in a subject by at least 10%, such as 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95% or more, compared to tumor angiogenesis in an untreated control subject or a subject receiving an antibody, a fragment thereof, or a polynucleotide encoding the antibody alone or a VEGF inhibitor alone before administration of an anti-MYCT1 antibody, a fragment thereof, or a polynucleotide encoding the antibody (baseline). Tumor angiogenesis in a subject can be assessed using any conventional method known in the art, non-limiting examples of which include diagnostic techniques, such as counting the number of blood vessels in a given area using a stereomicroscope or by measuring the expression of biomarkers associated with angiogenesis, including α v β3 integrin, endoglin and vascular endothelial growth factor (VEGF) receptor (VEGFR) 2.

[0357] In some aspects, administration of an anti-MYCT1 antibody, a fragment thereof, or a polynucleotide encoding the antibody in combination with a VEGF inhibitor compound can increase the immune response or anti-tumor immunity of a subject against cancer cells by at least 10%, for example, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or more, compared to the immune response or anti-tumor immunity of a control subject who has not received treatment or a subject who has received an antibody, a fragment thereof, or a polynucleotide encoding the antibody or a VEGF inhibitor alone before (baseline) administration of the antibody, a fragment thereof, or a polynucleotide encoding the antibody and a VEGF inhibitor. Biomarkers that reflect the intrinsic characteristics of the tumor immune microenvironment and tumor cells can be used to measure the immune response against cancer cells, such as PD-L1 expression and the density of tumor infiltrating lymphocytes (TILs).

[0358] In various aspects of the present disclosure, methods for regulating MYCT1 activity are provided. One aspect of the present disclosure provides methods for regulating tumor angiogenesis (anti-angiogenesis, targeting vascular control of Myct1) and / or immune stimulation in a subject, which inhibits tumor growth. In some aspects, the method includes administering an anti-MYCT1 antibody to a subject with cancer.

[0359] In other aspects, the methods disclosed herein encompass sensitizing the subject's cancer to immune checkpoint inhibitors. In this regard, the method includes selecting a subject with cancer, wherein the cancer is resistant to immune checkpoint inhibitors. In some aspects, the subject may be diagnosed by, for example, a physician as having cancer or suspected of having cancer or developing cancer, and the cancer is resistant or insensitive to immune checkpoint inhibitors. In other aspects, the method includes administering to the subject an effective amount of a composition comprising an anti-MYCT1 antibody disclosed herein or a fragment thereof. In some aspects, administering an anti-MYCT1 antibody or a fragment thereof in such subjects causes reduction of angiogenesis and increase of anti-tumor immunity in the subject. In other aspects, administering an anti-MYCT1 antibody in such subjects causes a reduction in tumor volume.

[0360] Further aspects include methods of modulating tumor angiogenesis and / or increasing anti-tumor immunity in a subject. In this aspect, the method comprises administering to the subject a therapeutically effective amount of a composition comprising an anti-MYCT1 antibody or fragment thereof.

[0361] In some aspects, the effective amount of an anti-MYCT1 antibody can be an amount that is substantially effective in blocking or inhibiting MYCT1 expression and / or activity in a cell. In some aspects, the effective amount of an anti-MYCT1 antibody can be an amount that is substantially effective in blocking or inhibiting MYCT1 expression and / or activity in endothelial cells (EC). In some aspects, blocking MYCT1 in endothelial cells promotes an immunostimulatory microenvironment by enhancing CTL infiltration and preventing CTL apoptosis; contributing to an immunostimulatory microenvironment; causing an anti-tumor microenvironment; and / or promoting endothelial regulation of tumor immunity. In some aspects, the effective amount of an anti-MYCT1 antibody can be an amount that effectively reduces angiogenesis. In some aspects, the effective amount of an anti-MYCT1 antibody can be an amount that effectively promotes the formation of high endothelial venules (HEV). In some aspects, administration of the composition of the present disclosure causes an increase in EC HEV. In some aspects, after administration of the anti-MYCT1 antibody of the present disclosure, the subject has improved anti-tumor drug delivery and enhanced anti-tumor immunity. In some aspects, the subject has high endothelial venule formation and an anti-tumor immune environment, causing limited tumor progression.

[0362] In some other aspects, the effective amount of anti-MYCT1 antibody can be an amount effective to enhance robust CTL infiltration. In some aspects, the effective amount of anti-MYCT1 antibody can be an amount effective to promote polarization of inflammatory M1 macrophages. In some aspects, administration of the compositions of the present disclosure causes reduced EC motility. In some aspects, administration of the compositions of the present disclosure causes defective tumor vasculature. In some aspects, administration of the compositions of the present disclosure causes downregulation of Fas ligand expression in EC.

[0363] In some aspects, the subject has upregulated MYCT1 in ECs prior to administration of the anti-MYCT1 antibody.Suitable compositions, administration forms, dosing, and dosing schedules for use in the methods of the present disclosure are disclosed herein, such as those described in Section III, which is incorporated by reference into this section.

[0364] In some aspects, the amount of anti-MYCT1 antibody and immune checkpoint inhibitor and / or VEGF inhibitor is an amount effective to reduce or prevent exhaustion of infiltrating cytotoxic T lymphocytes (CTLs). In some aspects, combined vascular and immune control provides synergistic anti-tumor activity.

[0365] In another aspect, the present disclosure provides a composition as disclosed herein for use in vitro, in vivo or ex vivo.

[0366] Therefore, in some aspects, the methods of the present disclosure can be used to reduce tumor or cancer progression or prolong the survival of subjects with cancer or tumors. The present disclosure provides that combined vascular and immune control provides synergistic anti-tumor activity. For example, using the anti-MYCT1 antibodies of the present disclosure to target MYCT1 improves the response to anti-PD1 therapy in treatment-responsive and treatment-refractory tumors, and combined MYCT1 and VEGF targeting with anti-PD1 therapy produces excellent tumor control, indicating a synergistic effect between the MYCT1 and VEGF pathways.

[0367] In one aspect, the method generally comprises the steps of: providing a composition described herein; and administering the composition to a subject or a cell (eg, an endothelial cell).

[0368] In some aspects, the present disclosure provides methods for treating tumors or cancer cells with increased risk of angiogenesis. In some aspects, the present disclosure provides methods for treating cancer or tumors by sequentially or simultaneously administering the compositions of the present disclosure with another cancer therapy. When sequentially administered, additional agents may be administered before the compounds of the present disclosure or before and after the compounds of the present disclosure. In preferred aspects, additional agents are immune checkpoint blockers and optional anti-VEGF therapy.

[0369] In other aspects, the compositions of the present disclosure can be delivered to cancer cells in vitro. The cancer cell can be a cancer cell line cultured in vitro. In some alternatives, the cancer cell line can be a primary cell line that has not yet been described. The method for preparing a primary cancer cell line utilizes standard techniques known to those skilled in the art. In other alternatives, the cancer cell line can be an established cancer cell line. The cancer cell line can be adherent or non-adherent, or the cell line can be grown under conditions that encourage adherent, non-adherent or organotypic growth using standard techniques known to those skilled in the art. The cancer cell line can be contact-inhibited or non-contact-inhibited.

[0370] In some aspects, the cancer cell line can be an established human cell line derived from a tumor. Non-limiting examples of cancer cell lines derived from tumors can include MM cell lines MM.1S, H929 and RPMI, osteosarcoma cell lines 143B, CAL-72, G-292, HOS, KHOS, MG-63, Saos-2 or U-2OS; prostate cancer cell lines DU145, PC3 or Lncap; breast cancer cell lines MCF-7, MDA-MB-438 or T47D; myeloid leukemia cell line THP-1, glioblastoma cell line U87; neuroblastoma cell line SHSY5Y; bone cancer cell line Saos-2; colon cancer cell lines WiDr, COLO 320DM, HT29, DLD-1, COLO205, COLO 201, HCT-15, SW620, LoVo, SW403, SW403, SW1116, SW1463, SW837, SW948, SW1417, GPC-16, HCT-8, HCT 116, NCI-H716, NCI-H747, NCI-H508, NCI-H498, COLO 320HSR, SNU-C2A, LS180, LS174T, MOLT-4, LS513, LS1034, LS411N, Hs 675.T, CO 88BV 59-1, Co88BV59H21-2, Co88BV59H21-2V67-66, 1116-NS-19-9, TA 99, AS 33, TS106, Caco-2, HT-29, SK-CO-1, SNU-C2B or SW480; B16-F10, RAW264.7, F8 cell lines or pancreatic cancer cell line Panc1.

[0371] In general, the methods as described herein comprise administering to a subject a therapeutically effective amount of a composition of the present disclosure. The methods described herein are generally performed on a subject in need thereof.

[0372] The level of MYCT1 can be used as a predictive marker for the angiogenic state of a subject, and thus can be used to make treatment decisions for subjects who may benefit from certain therapies (e.g., anti-angiogenesis).For example, the MYCT1 level of a candidate subject can be compared to a reference value.

[0373] Therefore, the present disclosure also provides anti-MYCT1 antibodies conjugated to a detectable signal (i.e., a measurable substance or a substance that generates a measurable signal). Non-limiting examples include various enzymes, prosthetic groups, fluorescent materials, luminescent materials, bioluminescent materials, radioactive materials, positron-emitting metals using various positron emission tomography, and non-radioactive paramagnetic metal ions. For example, see U.S. Patent No. 4,741,900 for metal ions, which can be conjugated to antibodies for use as diagnostic agents according to the present disclosure. Examples of suitable enzymes include horseradish peroxidase, alkaline phosphatase, β-galactosidase, or acetylcholinesterase; examples of suitable prosthetic group complexes include streptavidin / biotin and avidin / biotin; examples of suitable fluorescent materials include umbelliferone, fluorescein, fluorescein isothiocyanate, rhodamine, dichlorotriazinylamine fluorescein, dansyl chloride, or phycoerythrin; examples of luminescent materials include luminol; examples of bioluminescent materials include luciferase, luciferin, and aequorin; and examples of suitable radioactive materials include 125 I. 131 I. 111 In or 99 Tc. The signal generated by the agent can be measured, for example, by single photon emission computed tomography (SPECT) or positron emission tomography (PET).

[0374] Anti-MYCT1 antibodies conjugated to a detectable signal can be used diagnostically, for example, to monitor the development or progression of a tumor or cancer, as part of a clinical testing program, for example, to determine the efficacy of a given treatment and / or prevention regimen. Treatment of a patient can be adjusted based on the level of neurodegenerative disease progression.

[0375] In some aspects, a method for assessing disease progression in a subject being treated for cancer or a tumor is provided, the method comprising: (a) administering an anti-MYCT1 antibody disclosed herein labeled with an agent that generates a measurable signal as described herein (i.e., a "labeled anti-MYCT1 antibody"), wherein the signal is measured in the patient after administration; (b) administering a labeled anti-MYCT1 or antigen-binding fragment thereof at one or more time intervals after administration (a), wherein the signal is measured in the patient after administration; and (c) assessing the patient's disease progression based on changes in the signal measured in the patient at one or more time intervals after administration (a); wherein an increase in the signal indicates progression of the patient's neurodegenerative disease. In some aspects, the subject is being treated with the same anti-MYCT1 antibody, but in an unlabeled form. In some aspects, the subject is being treated with an anti-MYCT1 antibody that competitively inhibits the binding of the labeled anti-MYCT 1 antibody to human ApoE. In some aspects, the subject is being treated with an anti-MYCT1 antibody that does not competitively inhibit the binding of the labeled anti-MYCT1 antibody to human MYCT1. In some aspects, the subject is being treated with other drugs known in the art.

[0376] In other aspects, a method for evaluating disease progression in a subject being treated for cancer or a tumor is provided, the method comprising: (a) administering an anti-MYCT1 antibody disclosed herein labeled with an agent that generates a measurable signal as described herein (i.e., a "labeled anti-MYCT1 antibody"), wherein the signal is measured in the patient after administration; (b) assessing the subject's disease state after comparing the signal measured in the censored subject with the signal measured after administering the labeled antibody or antigen-binding fragment thereof to one or more control subjects; wherein an increase in the signal generated in the patient relative to the control subjects is associated with an increase in brain amyloid plaques; and (c) treating the patient with a therapy appropriate for the patient's disease state. "Control subject" refers to any normal healthy subject (or subject group), one or more subjects with varying degrees of disease, or even an actual test subject in the early stages of the disease.

[0377] Reference value can represent the same level of MYCT1 in control subject or represent MYCT1 level in control group.In some instances, the same level of MYCT1 in control subject or control group can be determined by the same method as for determining MYCT1 level in candidate subject.In some cases, control subject or control group can be a subject with cancer or tumor or a subject group with cancer or tumor that responds to treatment.In other cases, control subject or control group can be a subject with cancer or tumor that is unresponsive to treatment or a subject group with cancer or tumor.Alternatively, control subject or control group can refer to healthy subject or healthy subject group.In preferred aspects, control subject or control group belongs to the same species (such as human subject or human subject group) as candidate subject.As used herein, assessing "responsiveness" or "non-responsiveness" to therapy refers to determining the possibility that a subject responds to a therapeutic agent or does not respond to a therapeutic agent.For example, responsive subjects will have a therapeutic effect as a result of administering a medicament.In some aspects, therapeutic effect includes preventing or slowing down (mitigating) undesirable physiological changes or diseases / disorders. Beneficial or desired clinical results include, but are not limited to, alleviation of symptoms, reduction in extent of disease, stabilization of the disease state (i.e., not worsening), delay or slowing of disease progression, improvement or palliation of the disease state, and remission (whether partial or complete), or may also mean prolonged survival as compared to the expected survival of an untreated or non-responsive subject or population.

[0378] It should be understood that the methods provided herein do not require measuring reference values ​​each time a candidate subject is tested. On the contrary, in some aspects, it is contemplated that reference values ​​can be obtained and recorded, and any test level can be compared to such reference levels. The reference level can be a single cutoff value or a range of values.

[0379] By comparing the level of MYCT1 in a candidate subject as disclosed herein to a reference value also as described herein, the subject can be identified as responsive to therapy or likely to respond to therapy or non-responsive to therapy or unlikely to respond to therapy based on the assessment.

[0380] For example, when the reference value represents a level of MYCT1 that is responsive to a therapy, deviation from such a reference value will indicate non-responsiveness to the therapy. In another aspect, when the reference value represents a level of MYCT1 that is responsive to a genotoxic therapy, approximately the same phosphorylation level of MYCT1 in the candidate subject will indicate responsiveness to the therapy.

[0381] Alternatively, when the reference value represents a level of MYCT1 that is unresponsive to therapy, deviation from such reference value will indicate responsiveness to therapy. In another aspect, when the reference value represents a level of MYCT1 that is unresponsive to therapy, approximately the same level of MYCT1 in a candidate subject will indicate unresponsiveness to therapy.

[0382] In some cases, deviation means that the level of MYCT1 in the candidate subject (e.g., represented by a value) is increased or decreased relative to a reference value, such as at least 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 150%, 200%, 300%, 400%, 500% or more above or below a reference value.

[0383] Therefore, the present disclosure provides a method for evaluating responsiveness to therapy or detecting the angiogenic state of a cancer or tumor in a subject with cancer or a tumor. In one aspect, the method for evaluating responsiveness to therapy or detecting the angiogenic state may include (a) providing a biological sample obtained from a subject and measuring the level of MYCT1 in cancer cells or tumor cells or endothelial cells of the sample; and (b) when the measured MYCT 1 level deviates from a reference value, determining the responsiveness of the subject to therapy or angiogenic state.

[0384] MYCT1 levels can be used for a variety of mathematical operations to assess responsiveness or angiogenesis status. For example, MYCT1 values ​​can be combined with other known biomarkers for various statistical models (e.g., linear regression, LME curves, LOESS curves, etc.). The selection of measured values ​​and the selection of mathematical operations can be optimized to maximize the specificity of the method. For example, diagnostic accuracy can be evaluated by the area under the ROC curve, and in some aspects, a ROC AUC value of 0.7 or greater is set as a threshold value (e.g., 0.7, 0.75, 0.8, 0.85, 0.9, 0.95, etc.).

[0385] V. Kit

[0386] The present disclosure provides a kit for use with the methods of the present disclosure, comprising one or more anti-MYC T1 antibodies, fragments thereof, or polynucleotides encoding the antibodies. The kit may comprise a composition comprising an anti-MYCT1 antibody, a fragment thereof, or a polynucleotide encoding the antibody, or a pharmaceutical composition thereof, and instructions for administering the antibody, a fragment thereof, or a polynucleotide encoding the antibody, or a pharmaceutical composition thereof, to a subject in need thereof. The kit may further comprise an immune cell inhibitor and / or a VEGF inhibitor, which may be administered in combination with an anti-MYCT1 antibody, a fragment thereof, or a polynucleotide encoding the antibody. In some aspects, the kit may further comprise a sterile pharmaceutically acceptable carrier, buffer, or other diluent. The kit provided herein generally includes instructions for implementing the method. The instructions included in the kit may be attached to the packaging material or may be included as a package insert. Although the instructions are typically written or printed materials, they are not limited thereto. The present disclosure contemplates any medium capable of storing such instructions and communicating them to an end user. Such media include, but are not limited to, electronic storage media (e.g., disks, tapes, cassettes, chips), optical media (e.g., CD ROMs), and the like. As used herein, the term "instructions" may include the address of an Internet site that provides instructions.

[0387] The disclosed kits can have a single container containing the disclosed anti-MYCT1 antibodies, fragments thereof, or polynucleotides encoding the antibodies, with or without any additional components, or they can have different containers for various desired agents. In the case of providing combined therapy, a single solution can be premixed in molar equivalents or with one component in excess of another. Alternatively, the antibody, fragment thereof, or polynucleotide encoding the antibody and other anticancer agent components of the kit can be separately maintained in different containers prior to administration to the patient.

[0388] When the components of the kit are provided as one or more liquid solutions, the liquid solution is preferably an aqueous solution, wherein a sterile aqueous solution is particularly preferred. However, the components of the kit may be provided as dry powders. When the reagents or components are provided as dry powders, the powders may be reconstituted by adding a suitable solvent. It is contemplated that the solvent may also be provided in another container.

[0389] The containers of the kit generally include at least one vial, test tube, flask, bottle, syringe or other container means, into which the disclosed anti-MYCT1 antibody, fragment thereof or polynucleotide encoding the antibody and any other desired agent can be placed and preferably appropriately aliquoted. In the case of including separate components, the kit generally also contains a second vial or other container into which these are placed, so that separate designed doses can be administered. The kit may also include a second / third container means for containing a sterile pharmaceutically acceptable buffer or other diluent.

[0390] The kit may also contain a device by which the disclosed anti-MYCT1 antibodies, fragments thereof, or polynucleotides encoding the antibodies are administered to an animal or patient, such as one or more needles or syringes or even droppers, pipettes, or other such devices, from which the formulation can be injected into the animal or applied to the diseased area of ​​the body. The kit of the present invention typically also includes a device for containing vials or the like and other components that are tightly closed for commercial sale, such as injection or blow-molded plastic containers, into which the desired vials and other devices are placed and retained.

[0391] As various changes could be made in the materials and methods described above without departing from the scope of the invention, it is intended that all matter contained in the above description and in the examples given below shall be interpreted as illustrative and not in a limiting sense. DETAILED DESCRIPTION

[0392] Example

[0393] The following examples are included to demonstrate various aspects of the present disclosure. It will be appreciated by those skilled in the art that the techniques disclosed in the following examples represent techniques that the inventors have discovered to function well in the practice of the present invention, and therefore can be considered to constitute preferred modes for its practice. However, in view of the present disclosure, it will be appreciated by those skilled in the art that many changes may be made to the specific aspects disclosed without departing from the spirit and scope of the present invention, and still similar or similar results may be obtained.

[0394] Materials and methods

[0395] Table 2: Preparation of I20 medium

[0396]

[0397] Hybridoma culture

[0398] The culture medium shown in Table 3 was prepared using the following reagents.

[0399] Table 3: Culture media for hybridoma cell culture

[0400]

[0401]

[0402] The cells were approximately 1×10 5 cells / ml to 1×10 6 cells / ml were maintained in culture medium.

[0403] EC culture

[0404] MCEC and its derivatives: MCEC cells were prepared from FVB / N mouse hearts from mice with MHC haplotype "q". Culture medium was prepared using DMEM high glucose 10% FBS, 10mmol / L penicillin-streptomycin, and 10mmol / L HEPES. Gelatin pre-coating was recommended for this cell line, but not required. The cell doubling time was about 30 hours. Cells were subcultured at a subculture ratio of about 1:6. Cryopreservation of cells was performed at 1,500,000 cells / ml in FBS containing 10% DMSO.

[0405] MLEC, MLEC Myct1 KO, Myct1 KO MLEC overexpressing human short MYCT1 or long MYCT1, and their derivatives: iMLEC and iMLEC Myct1 KO cells were prepared from WT and Cdh5-cre Myct1 f / f C57 / B6 mouse lungs, respectively. Cells were immortalized by DOX-inducible SV40 T selection with puromycin and blasticidin (pLV[Tet]-TRE3G>SV40-T(ns):T2A:Puro and pLV[Exp]-Bsd-{mCd144}>Te t3 constructs). Culture medium was prepared using DMEM high glucose 10% FBS, 10mmol / L penicillin-streptomycin, 10mmol / L HEPES and DOX (40ng / ml). For optimal growth, 0.1% gelatin (in dH2O) coating of the culture vessel was required (30 minutes in a 37°C incubator>>absorb>>dry in a TC fume hood for 30 minutes>>ready to use). After passage, cells require about 3-4 days to completely refill the culture vessel. Subculture was performed at a split ratio of about 1:3. Cryopreservation was performed at 1,500,000 cells / ml using FBS containing 10% DMSO.

[0406] HUVEC, MYCT1 knockdown HUVEC, MYCT1 knockdown HUVEC overexpressing mouse Myct1:HUVEC cells were purchased from Fisher (Cat. No. 08-774-388). HUVEC cells were immortalized using human TERT lentivirus (vector: LB 12_FLAG_TERT_pCDH_MCS_EF1_NEO), followed by G418 selection. Endothelial cell growth medium (ready-to-use) (Promo Cell, Cat. No.: C-22010), a ready-to-use medium with about 2% FBS with additional serum to reach 10%, was used as a culture medium. For optimal growth, 0.1% gelatin (in dH2O) coating of the culture container is required (30 minutes in a 37°C incubator >> aspirate >> dry in a TC fume hood for 30 minutes >> ready to use). After passage, cells require about 4-5 days to completely refill the culture container. Passage was performed at a split ratio of 1:2. It was observed that HUVEC cells like to be close to each other, and if the cells are sparsely seeded or passaged, they may not grow at all. Cryopreservation was performed at 1,500,000 cells / ml in FBS containing 10% DMSO.

[0407] The HUVEC MYCT1 KD cell line was developed by transducing the HUVEC cell line with MYCT1 shRNA lentiviral particles, followed by puromycin selection. The HUVEC MYCT1 KD Myct1 overexpression line was generated by further transducing the iHUVEC MYCT1 KD cell line with Myct1 overexpression lentiviral particles, followed by blasticidin selection.

[0408] Endothelial tube formation assay

[0409] Endothelial cells were serum starved overnight. The next day, μ-Slide angiogenesis plates (Cat. No.: 81506, ibidi) were coated with Matrigel (Cat. No.: 96992, Corning) (10 μl / well) and incubated at 37°C for 30-45 minutes until Matrigel polymerized. Overnight serum-starved endothelial cells were plated on Matrigel-coated plates at 8,000 cells / well with 50 μl culture medium on top. The plates were incubated at 37°C for 6-8 hours before taking pictures with a microscope. The angiogenesis analyzer module of ImageJ software (NIH) was used to quantify the total tube length, the number of rings, and the number of branches.

[0410] Endothelial sprouting assay

[0411] The following reagents were used in the endothelial sprouting assay.

[0412] 1. EGM-2 Bullet Kit (containing FGF2 and VEGF-A; Lonza, catalog number CC-3162)

[0413] 2.M199

[0414] 3.DMEM high glucose

[0415] 4. Fetal bovine serum

[0416] 5. HBSS

[0417] 6. Dulbecco's phosphate buffered saline without Ca2+ and Mg2+ (DPBS; Life Technologies, Cat. No. 14190-094)

[0418] 7. Cytodex 3 microcarrier beads (GE Healthcare, cat. no. 17-0485-01)

[0419] 8. Fibrinogen type I from bovine plasma (Sigma-Aldrich, Cat. No. F8630)

[0420] 9. Thrombin from bovine plasma (Sigma-Aldrich, Catalog No. T3399)

[0421] 10. Aprotinin from bovine lung (Sigma-Aldrich, Cat. No. A1153)

[0422] 11. Trypsin / EDTA, UV inactivated (PAA Laboratories, Cat. No. L11-660)

[0423] 12.5ml, 15ml and 50ml polypropylene tubes

[0424] 13.5ml polystyrene tube (FACS tube)

[0425] 14. Low binding micro Eppendorf centrifuge tube (1.5ml)

[0426] 15.PKH kit

[0427] 16. 24-well glass bottom plate with #1.5 cover glass

[0428] 17. Cell counting kit

[0429] ...

Claims

1. An isolated anti-MYCT1 antibody or fragment thereof that specifically binds to an epitope within the first 100 amino acids of SEQ ID NO:

145.

2. The isolated anti-MYCT1 antibody or fragment thereof according to claim 1, comprising a light chain variable region and / or a heavy chain variable region, wherein the light chain variable region comprises L1 having an amino acid sequence of SEQ ID NO: 28, SEQ ID NO: 64, SEQ ID NO: 100 or SEQ ID NO: 136, L2 having an amino acid sequence of SEQ ID NO: 30, SEQ ID NO: 66, SEQ ID NO: 102 or SEQ ID NO: 138, L3 having an amino acid sequence of SEQ ID NO: 32, SEQ ID NO: 68, SEQ ID NO: 104 or SEQ ID NO: 140, or any combination thereof; the heavy chain variable region comprises H1 having an amino acid sequence of SEQ ID NO: 10, SEQ ID NO: 46, SEQ ID NO: 82 or SEQ ID NO: 118, H2 having an amino acid sequence of SEQ ID NO: 12, SEQ ID NO: 48, SEQ ID NO: 84 or SEQ ID NO: 120, H3 having an amino acid sequence of SEQ ID NO: 14, SEQ ID NO: 50, SEQ ID NO: 51 H3 of the amino acid sequence of SEQ ID NO:86 or SEQ ID NO:122, or any combination thereof.

3. The isolated anti-MYCT1 antibody or fragment thereof according to claim 1, comprising a light chain variable region and / or a heavy chain variable region, wherein the light chain variable region comprises the amino acid sequence of SEQ ID NO:36, SEQ ID NO:72, SEQ ID NO:108 or SEQ ID NO:144; the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:18, SEQ ID NO:54, SEQ ID NO:90 or SEQID NO:126, or any combination thereof.

4. An isolated anti-MYCT1 antibody or a fragment thereof, comprising a light chain variable region and / or a heavy chain variable region, wherein the light chain variable region comprises L1 having an amino acid sequence of SEQ ID NO: 28, L2 having an amino acid sequence of SEQ ID NO: 30, L3 having an amino acid sequence of SEQ ID NO: 32, or any combination thereof; the heavy chain variable region comprises H1 having an amino acid sequence of SEQ ID NO: 10, H2 having an amino acid sequence of SEQ ID NO: 12, H3 having an amino acid sequence of SEQ ID NO: 14, or any combination thereof.

5. An isolated anti-MYCT1 antibody or a fragment thereof, comprising a light chain variable region and / or a heavy chain variable region, wherein the light chain variable region comprises L1 having an amino acid sequence of SEQ ID NO: 64, L2 having an amino acid sequence of SEQ ID NO: 66, L3 having an amino acid sequence of SEQ ID NO: 68, or any combination thereof; the heavy chain variable region comprises H1 having an amino acid sequence of SEQ ID NO: 46, H2 having an amino acid sequence of SEQ ID NO: 48, H3 having an amino acid sequence of SEQ ID NO: 50, or any combination thereof.

6. An isolated anti-MYCT1 antibody or a fragment thereof, comprising a light chain variable region and / or a heavy chain variable region, wherein the light chain variable region comprises L1 having an amino acid sequence of SEQ ID NO: 100, L2 having an amino acid sequence of SEQ ID NO: 102, L3 having an amino acid sequence of SEQ ID NO: 104, or any combination thereof; the heavy chain variable region comprises H1 having an amino acid sequence of SEQ ID NO: 82, H2 having an amino acid sequence of SEQ ID NO: 84, H3 having an amino acid sequence of SEQ ID NO: 86, or any combination thereof.

7. An isolated anti-MYCT1 antibody or a fragment thereof, comprising a light chain variable region and / or a heavy chain variable region, wherein the light chain variable region comprises L1 having an amino acid sequence of SEQ ID NO: 136, L2 having an amino acid sequence of SEQ ID NO: 138, L3 having an amino acid sequence of SEQ ID NO: 140, or any combination thereof; the heavy chain variable region comprises H1 having an amino acid sequence of SEQ ID NO: 118, H2 having an amino acid sequence of SEQ ID NO: 120, H3 having an amino acid sequence of SEQ ID NO: 122, or any combination thereof.

8. An isolated anti-MYCT1 antibody or a fragment thereof, comprising the amino acid sequence of the light chain variable region of SEQ ID NO: 36 and / or the amino acid sequence of the heavy chain variable region of SEQ ID NO:

18.

9. An isolated anti-MYCT1 antibody or a fragment thereof, comprising the amino acid sequence of the light chain variable region of SEQ ID NO: 72 and / or the amino acid sequence of the heavy chain variable region of SEQ ID NO:

54.

10. An isolated anti-MYCT1 antibody or a fragment thereof, comprising the amino acid sequence of the light chain variable region of SEQ ID NO: 108 and / or the amino acid sequence of the heavy chain variable region of SEQ ID NO:

90.

11. An isolated anti-MYCT1 antibody or a fragment thereof, comprising the amino acid sequence of the light chain variable region of SEQ ID NO: 144 and / or the amino acid sequence of the heavy chain variable region of SEQ ID NO:

126.

12. The isolated antibody or fragment thereof according to any one of the preceding claims, wherein the antibody is a monoclonal antibody.

13. The isolated antibody or fragment thereof according to any one of the preceding claims, wherein the antibody is a humanized antibody, a single chain variable fragment (scFv) antibody, an antibody fragment or a chimeric antibody.

14. A pharmaceutical composition comprising the isolated antibody or fragment thereof according to any one of the preceding claims and a pharmaceutically acceptable carrier or excipient.

15. An isolated antibody according to any preceding claim for use as a medicament.

16. A method of treating cancer in a subject in need thereof, the method comprising: An effective amount of a composition comprising the isolated antibody or fragment thereof according to any one of the preceding claims is administered.

17. The method according to claim 16, further comprising: Administer immune checkpoint inhibitors and optional anti-VEGF targeted therapy.

18. A method of sensitizing a subject's cancer to an immune checkpoint inhibitor, the method comprising: selecting a subject having a cancer wherein the cancer is resistant to an immune checkpoint inhibitor; and An effective amount of a composition comprising the isolated antibody or fragment thereof according to any one of claims 1-15 is administered to the subject.

19. The method of any one of claims 16-18, wherein administration of the anti-MYCT1 antibody results in reduced angiogenesis of cancer cells in the subject.

20. The method of any one of claims 16-18, wherein administration of an anti-MYCT1 antibody results in enhanced anti-tumor immunity in the subject.

21. The method of any one of claims 16-18, wherein administration of the anti-MYCT1 antibody results in a reduction in tumor volume in the subject.

22. A method for regulating tumor angiogenesis and / or enhancing anti-tumor immunity in a subject suffering from cancer, the method comprising: A therapeutically effective amount of a composition comprising the isolated antibody or fragment thereof according to any one of claims 1-15 is administered to the subject.

23. The method of claim 27, further comprising: Administer an immune checkpoint inhibitor and optionally a VEGF inhibitor.

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