FC-null Anti-GIP antibodies

Abstract

The disclosure provides Fc polypeptides wherein the Fc domain comprises at least one amino acid substitution that reduces binding affinity to an Fc receptor and / or effector function comprising one or more, two or more, three or more, four or more, or all five amino acid substitutions selected from L234A, L235A, G237A, K322A, and P329G, wherein the residues are numbered according to the EU index, preferably wherein the Fc polypeptide is an antibody. In some embodiments, the antibody is an anti-GIP antibody. In some embodiments, the antibody further comprises the three M252Y, S254T and T256E substitutions or the two M428L and N434S substitutions, which extend the half-life of the antibody.

Description

Fc-Null Anti-GIP AntibodiesField of the Invention

[0001] The disclosure provides Fc polypeptides wherein the Fc domain comprises at least one amino acid substitution that reduces binding affinity to an Fc receptor and / or effector function comprising one or more, two or more, three or more, four or more, or all five amino acid substitutions selected from L234A, L235A, G237A, K322A, and P329G, wherein the residues are numbered according to the EU index, preferably wherein the Fc polypeptide is an antibody. In some embodiments, the antibody is an anti-GIP antibody. The disclosure provides anti-GIP antibodies with reduced effector functions and methods of using the same to treat obesity and obesity-associated diseases or disorders.Background

[0002] Obesity is a chronic, relapsing, multifactorial disease whose prevalence continues to increase worldwide. It is estimated that there are about 600 million obese adults worldwide with Body Mass Index (BMI) > 30 kg / m2. In the United States alone, 69% of adults are overweight or obese. Estimated costs to the healthcare system are more than $550 billion annually. Increased severity of obesity correlates with a higher prevalence of the associated comorbidities. Likewise, obesity increases the risk of premature mortality. Obesity affects almost every organ system in the body and increases the risk of numerous diseases including type 2 diabetes mellitus, hypertension, dyslipidemia, cardiovascular disease, and cancer. It is estimated that a man in his twenties with a BMI over 45 will have a 22% reduction (13 years) in life expectancy.

[0003] Obesity has been linked to the function of gastric inhibitory polypeptide (GIP), which is a 42-amino-acid hormone secreted from K cells of the upper small intestine. It was originally isolated from porcine intestine on the basis of its ability to inhibit gastric acid secretion. Later, it was found that GIP administration stimulates insulin secretion in healthy volunteers, and that GIP acts directly on pancreatic islets to stimulate insulin secretion. These lines of evidence showed GIP to be the first incretin, which was then renamed glucose-dependent insulinotropic polypeptide. GIP exerts its effects by binding to its specific receptors, the GIP receptor (GIPR). GIPRs are distributed in the pancreas as well as in extra-pancreatic tissues, such as adiposetissue, the heart, the pituitary, the adrenal cortex, and some areas of the central nervous system (CNS). GIP has long been considered a hormone that promotes obesity. Incretins also regulate gastric motility, nutrient absorption, blood flow, and food intake. GIP is excessively secreted after nutrient consumption, and it promotes fat deposition in adipose tissue. However, the role of GIP in weight control is not clear. Both agonism and antagonism of the GIPR can reduce body weight in response to overnutrition in preclinical models. The remarkable protection against diet- induced obesity originally reported in Gipr— / — mice sparked a number of efforts to antagonize the GIPR pharmacologically. The other incretin is glucagon-like peptide 1 (GLP-1), produced by the L cells in the distal gut, which also stimulates insulin secretion in p cells through activation of the GLP-1 receptor (GLP-1 R). Several studies have demonstrated that GLP-1 R agonists are effective in treating or preventing obesity. Enhanced sensitivity to GLP-1 R agonists was also seen following chronic pharmacological antagonism of the GIPR.Summary of the Invention

[0004] The present disclosure provides, among other things anti-GIP antibodies comprising Fc variants that have significantly reduced ADCC, ADCP and CDC function. As described herein, the present disclosure is, in part based on identification of novel combinations of mutations that abolish binding to all FcyRI, FcyRlla, FcyRI I b, FcyRI I la, FcyRI 11 b, and C1q.

[0005] In one embodiment, the antibody has a human IgG constant region wherein the Fc domain comprises at least one amino acid substitution that reduces binding affinity to an Fc receptor and / or effector function. In some embodiments, an Fc variant comprises one or more amino acid substitutions selected from L234A, L235A, G237A, K322A, and P329G, wherein the residues are numbered according to the EL) index. In some embodiments, an Fc variant comprises the amino acid substitution L234A. In some embodiments, an Fc variant comprises the amino acid substitution L235A. In some embodiments, an Fc variant comprises the amino acid substitution G237A. In some embodiments, an Fc variant comprises the amino acid substitution K322A. In some embodiments, an Fc variant comprises the amino acid substitution P329G.

[0006] Non-limiting embodiments of the disclosure include:

[0007] We will list all claims here as ’’embodiments” when complete.

[0008] Any embodiment or embodiment described herein may be combined with any other embodiment or embodiment as disclosed herein. While the present disclosure has been described in conjunction with the detailed description thereof, the foregoing description isintended to illustrate and not limit the scope of the present disclosure, which is defined by the scope of the appended claims, other embodiments, advantages, and modifications are within the scope of the following embodiments.Brief Description of the Figures

[0009] The patent or application file contains at least one drawing executed in color.Copies of this patent or patent application publication with color drawings will be provided by the U.S. Patent and Trademark Office upon request and payment of the necessary fee.

[0010] FIG. 1 : SEC-HPLC profile of lncre001 and lncre002 after protein A purification showing both protein is 100% pure and the absence of aggregation.

[0011] FIG. 2: Sensograms of lncre001 (FIG. 2A) and lncre002 (FIG. 2B) showing interaction with the target, human GIP. Data fitted to the 1 :1 kinetic model shows similar KD values of 6.2 and 8 nM respectively

[0012] FIG. 3: Sensograms of lncre001 and lncre002 interactions with (FIG. 3A) human FcyRI, (FIG. 3B) human FcyRlla, (FIG. 3C) human FcyRllb and (FIG. 3D) human FcyRIIH .Unlike lncre001 , lncre002 has no binding to all the human FcyRs

[0013] FIG. 4: Sensograms of lncre001 (FIG. 4A) and lncre002 (FIG. 4B) showing interaction with the target, human GIP peptide. Data fitted to the 1 :1 kinetic model shows similar KD values of 0.4 and 1.2 nM, respectively.

[0014] FIG. 5: 13 peptides were synthesized, some containing alanine mutations, to map out the epitopes of lncre001 and lncre002. FIG. 5 discloses SEQ ID NOS 17-30, respectively, in order of appearance.

[0015] FIG. 6: A plot of OD value (450 nm - 620 nm) against a series of synthesized peptides to map out the binding epitopes of lncre001 and lncre002.

[0016] FIG. 7: A table showing the OD value (450 nm - 620 nm) against a series of synthesized peptides, some containing alanine mutations to map out lncre001 and lncre002 epitopes. FIG. 7 discloses SEQ ID NOS 17-30, respectively, in order of appearance.

[0017] FIG. 8: SEC-HPLC profile of lncre004 and lncre005 after protein A purification showing both protein is 100% pure and the absence of aggregation.

[0018] FIG. 9: Sensograms of lncre004, lncre005 and lncre002 showing interaction wit human FcRn at pH 6.0. Data fitted to the 1 :1 kinetic model shows lncre004 and lncre005 exhibited slower off rates than lncre002.

[0019] FIG. 10: Table showing Kd values of lncre004, lncre005 and lncre002 at pH 6.0 and 7.4.

[0020] FIG. 11 : Sensogram of lncre003 showing interaction with the target, mouse GIP peptide. Data fitted to the 1 :1 kinetic model shows KD value of 1.24 nM.

[0021] FIG. 12: mlgG2a control (top left) shows binding to all the 4 different mouse Fey receptors at 333 nM but lncre003 does not show any measurable binding to the different mouse Fcv receptors from 1000 nM tp 4.11 nMDetailed Description

[0022] DEFINITIONS

[0023] In order for the present disclosure to be more readily understood, certain terms are first defined below. Additional definitions for the following terms and other terms are set forth throughout the Specification.

[0024] As used in this Specification and the appended claims, the singular forms “a,” “an” and “the” include plural referents unless the context clearly dictates otherwise.

[0025] Unless specifically stated or obvious from context, as used herein, the term “or” is understood to be inclusive and covers both “or” and “and”.

[0026] The term “and / or” where used herein is to be taken as specific disclosure of each of the two specified features or components with or without the other. Thus, the term “and / or” as used in a phrase such as “A and / or B” herein is intended to include A and B; A or B; A (alone); and B (alone). Likewise, the term “and / or” as used in a phrase such as “A, B, and / or C” is intended to encompass each of the following aspects: A, B, and C; A, B, or C; A or C; A or B; B or C; A and C; A and B; B and C; A (alone); B (alone); and C (alone).

[0027] The terms “e.g.,” and “i.e.,” as used herein, are used merely by way of example, without limitation intended, and should not be construed as referring only those items explicitly enumerated in the specification.

[0028] The terms “or more,” “at least,” “more than,” and the like, e.g., “at least one” are understood to include but not be limited to at least 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 , 12, 13, 14, 15, 16, 17, 18, 19 20, 21 , 22, 23, 24, 25, 26, 27, 28, 29, 30, 31 , 32, 33, 34, 35, 36, 37, 38, 39, 40, 41 , 42, 43, 44, 45, 46, 47, 48, 49, 50, 51 , 52, 53, 54, 55, 56, 57, 58, 59, 60, 61 , 62, 63, 64, 65,66, 67, 68, 69, 70, 71 , 72, 73, 74, 75, 76, 77, 78, 79, 80, 81 , 82, 83, 84, 85, 86, 87, 88, 89, 90,91 , 92, 93, 94, 95, 96, 97, 98, 99, 100, 101 , 102, 103, 104, 105, 106, 107, 108, 109, 110, 111 ,112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130,131 , 132, 133, 134, 135, 136, 137, 138, 139, 140, 141 , 142, 143, 144, 145, 146, 147, 148, 149 or 150, 200, 300, 400, 500, 600, 700, 800, 900, 1000, 2000, 3000, 4000, 5000, or more than the stated value. Also included is any greater number or fraction in between.

[0029] Conversely, the term “no more than” includes each value less than the stated value. For example, “no more than 100 amino acids” includes 100, 99, 98, 97, 96, 95, 94, 93, 92, 91 , 90, 89, 88, 87, 86, 85, 84, 83, 82, 81, 80, 79, 78, 77, 76, 75, 74, 73, 72, 71, 70, 69, 68,67, 66, 65, 64, 63, 62, 61 , 60, 59, 58, 57, 56, 55, 54, 53, 52, 51 , 50, 49, 48, 47, 46, 45, 44, 43,42, 41 , 40, 39, 38, 37, 36, 35, 34, 33, 32, 31, 30, 29, 28, 27, 26, 25, 24, 23, 22, 21, 20, 19, 18,17, 16, 15, 14, 13, 12, 11 , 10, 9, 8, 7, 6, 5, 4, 3, 2, 1 , and 0 amino acids. Also included is any lesser number or fraction in between.

[0030] The terms “plurality,” “at least two,” “two or more,” “at least second,” and the like, are understood to include but not limited to at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 20, 21 , 22, 23, 24, 25, 26, 27, 28, 29, 30, 31 , 32, 33, 34, 35, 36, 37, 38, 39, 40, 41 , 42, 43, 44, 45, 46, 47, 48, 49, 50, 51 , 52, 53, 54, 55, 56, 57, 58, 59, 60, 61 , 62, 63, 64, 65, 66, 67, 68, 69, 70, 71 , 72, 73, 74, 75, 76, 77, 78, 79, 80, 81 , 82, 83, 84, 85, 86, 87, 88, 89, 90, 91 , 92, 93, 94, 95, 96, 97, 98, 99, 100, 101 , 102, 103, 104, 105, 106, 107, 108, 109, 110, 111 , 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131 , 132, 133, 134, 135, 136, 137, 138, 139, 140, 141 , 142, 143, 144, 145, 146, 147, 148, 149 or 150, 200, 300, 400, 500, 600, 700, 800, 900, 1000, 2000, 3000, 4000, 5000, or more. Also included is any greater number or fraction in between.

[0031] Throughout the specification the word “comprising,” or variations such as “comprises” or “comprising,” will be understood to imply the inclusion of a stated element, integer or step, or group of elements, integers or steps, but not the exclusion of any other element, integer or step, or group of elements, integers or steps. It is understood that wherever aspects are described herein with the language “comprising,” otherwise analogous aspects described in terms of “consisting of” and / or “consisting essentially of” are also provided. The term "consisting of" excludes any element, step, or ingredient not specified in the claim. In re Gray, 53 F.2d 520, 11 USPQ 255 (CCPA 1931); Ex parte Davis, 80 USPQ 448, 450 (Bd. App. 1948) ("consisting of" defined as "closing the claim to the inclusion of materials other than those recited except for impurities ordinarily associated therewith"). The term “consisting essentially of” limits the scope of a claim to the specified materials or steps "and those that do not materially affect the basic and novel characteristic(s)" of the claimed disclosure.

[0032] Unless specifically stated or evident from context, as used herein, the term “about” refers to a value or composition that is within an acceptable error range for the particular value or composition as determined by one of ordinary skill in the art, which will depend in part on how the value or composition is measured or determined, i.e., the limitations of the measurement system. For example, “about” or “approximately” may mean within one or more than one standard deviation per the practice in the art. “About” or “approximately” may mean a range of up to 10% (i.e., ±10%). Thus, “about” may be understood to be within 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1 %, 0.5%, 0.1%, 0.05%, 0.01%, or 0.001% greater or less than the stated value. For example, about 5 mg may include any amount between 4.5 mg and 5.5 mg. Furthermore, particularly with respect to biological systems or processes, the terms may mean up to an order of magnitude or up to 5-fold of a value. When particular values or compositions are provided in the instant disclosure, unless otherwise stated, the meaning of “about” or “approximately” should be assumed to be within an acceptable error range for that particular value or composition.

[0033] As described herein, any concentration range, percentage range, ratio range or integer range is to be understood to be inclusive of the value of any integer within the recited range and, when appropriate, fractions thereof (such as one-tenth and one-hundredth of an integer), unless otherwise indicated.

[0034] Units, prefixes, and symbols used herein are provided using their Systeme International de Unites (SI) accepted form. Numeric ranges are inclusive of the numbers defining the range.

[0035] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure is related. For example, Juo, “The Concise Dictionary of Biomedicine and Molecular Biology”, 2nd ed., (2001), CRC Press; “The Dictionary of Cell & Molecular Biology”, 5th ed., (2013), Academic Press; and “The Oxford Dictionary Of Biochemistry And Molecular Biology”, Cammack et al. eds., 2nd ed, (2006), Oxford University Press, provide those of skill in the art with a general dictionary for many of the terms used in this disclosure.

[0036] A “therapeutically effective amount,” “effective dose,” “effective amount,” or “therapeutically effective dosage” of a therapeutic agent, e.g., antibodies, small molecules, “agents” described in the specification, is any amount that, when used alone or in combination with another therapeutic agent, protects a subject against the onset of a disease or promotes disease regression evidenced by a decrease in severity of disease symptoms, an increase infrequency and duration of disease symptom-free periods, or a prevention of impairment or disability due to the disease affliction. Such terms may be used interchangeably. The ability of a therapeutic agent to promote disease regression may be evaluated using a variety of methods known to the skilled practitioner, such as in human subjects during clinical trials, in animal model systems predictive of efficacy in humans, or by assaying the activity of the agent in in vitro assays. Therapeutically effective amounts and dosage regimens can be determined empirically by testing in known in vitro or in vivo (e.g., animal model) systems.

[0037] The term "combination" refers to either a fixed combination in one dosage unit form, or a combined administration where a compound of the present disclosure and a combination partner (e.g., another drug as explained below, also referred to as "therapeutic agent" or "agent") may be administered independently at the same time or separately within time intervals, especially where these time intervals allow that the combination partners show a cooperative, e.g., synergistic effect. The single components may be packaged in a kit or separately. One or both of the components (e.g., powders or liquids) may be reconstituted or diluted to a desired dose prior to administration. The terms "co-administration" or "combined administration" or the like as utilized herein are meant to encompass administration of the selected combination partner to a single subject in need thereof (e.g., a patient), and are intended to include treatment regimens in which the agents are not necessarily administered by the same route of administration or at the same time.

[0038] The terms “homologous,” “homology,” or “percent homology” as used herein refer to the degree of sequence identity between an amino acid or polynucleotide sequence and a corresponding reference sequence. “Homology” can refer to polymeric sequences, e.g., polypeptide or DNA sequences that are similar. Homology can mean, for example, nucleic acid sequences with at least about: 50%, 55%, 60%, 65%, 70%, 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity. In other embodiments, a “homologous sequence” of nucleic acid sequences may exhibit 93%, 95%, or 98% sequence identity to the reference nucleic acid sequence. For example, a “region of homology to a genomic region” can be a region of DNA that has a similar sequence to a given genomic region in the genome. A region of homology can be of any length that is sufficient to promote binding of a spacer or protospacer sequence to the genomic region. For example, the region of homology can comprise at least 5, 10, 11 , 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 , 22, 23, 24, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 200, 300, 400, 500, 600, 700, 800, 900, 1000, 1100, 1200, 1300, 1400, 1500, 1600, 1700, 1800, 1900,2000, 2100, 2200, 2300, 2400, 2500, 2600, 2700, 2800, 2900, 3000, 3100, or more bases in length such that the region of homology has sufficient homology to undergo binding with the corresponding genomic region. When a percentage of sequence homology or identity is specified, in the context of two nucleic acid sequences or two polypeptide sequences, the percentage of homology or identity generally refers to the alignment of two or more sequences across a portion of their length when compared and aligned for maximum correspondence. When a position in the compared sequence can be occupied by the same base or amino acid, then the molecules can be homologous at that position. Unless stated otherwise, sequence homology or identity is assessed over the specified length of the nucleic acid, polypeptide, or portion thereof. In some embodiments, the homology or identity is assessed over a functional portion or a specified portion of the length. Alignment of sequences for assessment of sequence homology can be conducted by algorithms known in the art, such as the Basic Local Alignment Search Tool (BLAST) algorithm, which is described in Altschul et al, J. Mol. Biol.215:403- 410, 1990. A publicly available, internet interface, for performing BLAST analyses is accessible through the National Center for Biotechnology Information. Additional known algorithms include those published in: Smith & Waterman, “Comparison of Biosequences”, Adv. Appl. Math.2:482, 1981 ; Needleman & Wunsch, “A general method applicable to the search for similarities in the amino acid sequence of two proteins” J. Mol. Biol.48:443, 1970; Pearson & Lipman “Improved tools for biological sequence comparison”, Proc. Natl. Acad. Sci. USA 85:2444, 1988; or by automated implementation of these or similar algorithms. Global alignment programs may also be used to align similar sequences of roughly equal size. Examples of global alignment programs include NEEDLE (available at www.ebi.ac.uk / Tools / psa / emboss_needle / ) which is part of the EMBOSS package (Rice P et al., Trends Genet., 2000; 16: 276-277), and the GGSEARCH program fasta.bioch.virginia.edu / fasta_www2 / , which is part of the PASTA package (Pearson W and Lipman D, 1988, Proc. Natl. Acad. Sci. USA, 85: 2444-2448). Both of these programs are based on the Needleman- Wunsch algorithm, which is used to find the optimum alignment (including gaps) of two sequences along their entire length. A detailed discussion of sequence analysis can also be found in Unit 19.3 of Ausubel et al ("Current Protocols in Molecular Biology" John Wiley & Sons Inc, 1994-1998, Chapter 15, 1998). A skilled person understands that amino acid (or nucleotide) positions may be determined in homologous sequences based on alignment.

[0039] A “patient” or a “subject” as used herein includes any human who is afflicted with a heart disease or disorder. The terms “subject” and “patient” are used interchangeably herein.

[0040] As used herein, the term “in vitro cell” refers to any cell which is cultured ex vivo. In particular, an in vitro cell may include a T cell. The term “in vivo” means within the patient.

[0041] The term “anti-GIP” (Ab) includes, without limitation, a glycoprotein immunoglobulin, which binds specifically to the GIP antigen. In general, an antibody may comprise at least two heavy (H) chains and two light (L) chains interconnected by disulfide bonds, or an antigen-binding molecule thereof. Each H chain comprises a heavy chain variable region (abbreviated herein as VH) and a heavy chain constant region. The heavy chain constant region comprises three constant domains, CH1, CH2, and CH3. Each light chain comprises a light chain variable region (abbreviated herein as VL) and a light chain constant region. The light chain constant region is comprises one constant domain, CL. The VH and VL regions may be further subdivided into regions of hypervariability, termed complementarity determining regions (CDRs), interspersed with regions that are more conserved, termed framework regions (FR). Each VH and VL comprises three CDRs and four FRs, arranged from amino-terminus to carboxy-terminus in the following order: FR1 , CDR1 , FR2, CDR2, FR3, CDR3, and FR4. The variable regions of the heavy and light chains contain a binding domain that interacts with an antigen. The constant regions of the Abs may mediate the binding of the immunoglobulin to host tissues or factors, including various cells of the immune system (e.g., effector cells) and the first component (C1q) of the classical complement system. In a specific set of examples of this application, the anti-GIP antibody is NM-136.

[0042] Antibodies may include, for example, monoclonal antibodies, recombinantly produced antibodies, monospecific antibodies, multispecific antibodies (including bispecific antibodies), human antibodies, engineered antibodies, humanized antibodies, chimeric antibodies, immunoglobulins, synthetic antibodies, tetrameric antibodies comprising two heavy chain and two light chain molecules, an antibody light chain monomer, an antibody heavy chain monomer, an antibody light chain dimer, an antibody heavy chain dimer, an antibody light chainantibody heavy chain pair, intrabodies, antibody fusions (sometimes referred to herein as “antibody conjugates”), heteroconjugate antibodies, single domain antibodies, monovalent antibodies, single chain antibodies or single-chain Fvs (scFv), camelized antibodies, affybodies, Fab fragments, F(ab’)2 fragments, disulfide-linked Fvs (sdFv), anti-idiotypic (anti-ld) antibodies (including, e.g., anti-anti-ld antibodies), minibodies, domain antibodies, synthetic antibodies (sometimes referred to herein as “antibody mimetics”), and antigen-binding fragments of any of the above. In certain embodiments, antibodies described herein refer to polyclonal antibody populations.

[0043] An immunoglobulin may derive from any of the commonly known isotypes, including but not limited to IgA, secretory IgA, IgG and IgM. IgG subclasses are also well known to those in the art and include but are not limited to human lgG1 , lgG2, lgG3 and lgG4. “Isotype” refers to the Ab class or subclass (e.g., IgM or IgG 1 ) that is encoded by the heavy chain constant region genes. The term “antibody” includes, by way of example, both naturally occurring and non-naturally occurring Abs; monoclonal and polyclonal Abs; chimeric and humanized Abs; human or nonhuman Abs; wholly synthetic Abs; and single chain Abs. A nonhuman Ab may be humanized by recombinant methods to reduce its immunogenicity in man. Where not expressly stated, and unless the context indicates otherwise, the term “antibody” also includes an antigen-binding fragment or an antigen-binding portion of any of the aforementioned immunoglobulins, and includes a monovalent and a divalent fragment or portion, and a single chain Ab.

[0044] An “antigen binding molecule”, “antigen binding protein” “antigen binding portion / fragment,” or “antibody fragment” refers to any molecule that comprises the antigen binding parts (e.g., CDRs) of the antibody from which the molecule is derived. In this application, the antigen is GIP. An antigen binding molecule may include the antigenic complementarity determining regions (CDRs). Examples of antibody fragments include, but are not limited to, Fab, Fab', F(ab')2, and Fv fragments, dAb, linear antibodies, scFv antibodies, and multispecific antibodies formed from antigen binding molecules. Peptibodies (i.e., Fc fusion molecules comprising peptide binding domains) are another example of suitable antigen binding molecules. In some embodiments, the antigen binding molecule binds to an epitope present in the surface of a viral particle. In some embodiments, the antigen binding molecule binds to an epitope in GIP. In certain embodiments, the antigen binding molecule binds to any one of polypeptides of the disclosure. In further embodiments, the antigen binding molecule is an antibody fragment that specifically binds to the antigen, including one or more of the complementarity determining regions (CDRs) thereof. In further embodiments, the antigen binding molecule is a single chain variable fragment (scFv). In some embodiments, the antigen binding molecule comprises or consists of avimers.

[0045] The terms "isolated antibody" and “isolated peptide” refer to a protein or peptide produced from cDNA-, recombinant RNA-, or any other synthetic origin, or some combination thereof; as well as to proteins and peptides that, by virtue of their origin, or source of derivation, either (1) are not associated with proteins found in nature, (2) are free of other proteins from thesame source, e.g. free of murine proteins, (3) are expressed by a cell from a different species, or (4) do not occur in nature.

[0046] The term “epitope” is used here to refer to binding sites recognized by a binding protein or an antibody. Epitopes may be any molecule or grouping thereof, including, but not limited to, amino acids, amino acid side chains, sugars, and lipids, and may have a specific three-dimensional structure or conformation. Thus, an epitope may comprise any portion of GIP that includes primary, secondary, tertiary, or quaternary structure, as those terms are generally known in the art. A “linear epitope” is made up of a continuous sequence of amino acid residues. A “conformational epitope” is an epitope to which the antibody or binding protein binds in a conformational-specific manner. In the case of protein-based epitopes, the binding may depend on the epitope-carrying-protein’s secondary, tertiary, or quaternary structure. In other words, the antibody binds in a structure-specific manner, a tertiary-structure-specific manner, or a quaternary-structure-specific manner.

[0047] ’’Binding affinity” generally refers to the strength of the sum total of non-covalent interactions between a single binding site of a molecule (e.g., an antibody) and its binding partner (e.g., an antigen). Unless indicated otherwise, as used herein, “binding affinity” refers to intrinsic binding affinity that reflects a 1 :1 interaction between members of a binding pair (e.g., antibody and antigen). The affinity of a molecule X for its partner Y can generally be represented by the dissociation constant (Kd). Affinity can be measured by common methods known in the art, including those described herein. Low-affinity antibodies generally bind antigen slowly and tend to dissociate readily, whereas high-affinity antibodies generally bind antigen faster and tend to remain bound longer. A variety of methods of measuring binding affinity are known in the art, any of which can be used for purposes of the present disclosure. Specific illustrative and exemplary embodiments for measuring binding affinity are described in the Examples section.

[0048] The phrase "specifically bind(s)" or "bind(s) specifically" when referring to a binding molecule refers to a binding molecule which has intermediate or high binding affinity, exclusively or predominately, to a target molecule, such as to GIP or a cytokine receptor. The phrase "specifically binds to" refers to a binding reaction which is determinative of the presence of a target protein (such as GIP or a cytokine receptor) in the presence of a heterogeneous population of proteins and other biologies. Thus, under designated assay conditions, the specified binding molecules bind preferentially to a particular target protein (e.g. GIP) and do not bind in a significant amount to other components present in a test sample. Specific binding to atarget protein under such conditions may require a binding molecule that is selected for its specificity for a particular target protein. A variety of assay formats may be used to select binding molecules that are specifically reactive with a particular target protein. For example, solid-phase ELISA immunoassays, immunoprecipitation, Biacore and Western blot may be used to identify binding molecules that specifically bind to GIP. Typically, a specific or selective reaction will be at least twice background signal or noise and more typically more than 10 times background. Given that the binding molecule is an antibody, the phrase "specifically binds to" refers to a binding reaction that is determinative of the presence of the antigen (such as GIP) in a heterogeneous population of proteins and other biologies. Typically, an agent that specifically binds to an antigen binds the antigen with a dissociation constant (KD) of at least about 1 x 10’6to 1x 10-7, or about 1x 10-8to 1x 10’9M, or about 1x10-10to 1x10-11or higher; and / or binds to the predetermined antigen (e.g. of GIP) with an affinity that is at least two-fold, five-fold, ten-fold, twenty-fold greater than its affinity for binding to a non-specific antigen (e.g., BSA, casein) other than the predetermined antigen or a closely- related antigen. According to a specific embodiment, the affinity of the anti-GIP antibody for GIP is in the range of 10’8M-10’14M, such as determined by a surface plasmon resonance (SPR) assay. According to some embodiments, the affinity range is 10’8M-10’14M. According to some embodiments, the affinity range is 10’8M- 10’13M. According to some embodiments, the affinity range is 10’8M-10’12M. According to some embodiments, the affinity range is 10’8M-10’11M. According to some embodiments, the affinity range is 10-8M-10-10M. According to some embodiments, the affinity range is 10’8M-10’19M.

[0049] According to some embodiments, the affinity range is 10’9M-10’14M. According to some embodiments, the affinity range is 10’9M-10’13M. According to some embodiments, the affinity range is 10’9M-10’12M. According to some embodiments, the affinity range is 10’9M-10’11M. According to some embodiments, the affinity range is 10’9M-10’10M. According to some embodiments, the affinity range is 10’10M-10’13M. According to some embodiments, the affinity range is 10’1°M-10’12M. According to some embodiments, the affinity range is 10’1°M-10’11M.

[0050] Affinity is determined using a variety of techniques, an example of which is an affinity ELISA assay. In various embodiments, affinity is determined by a surface plasmon resonance assay (e.g., BIAcore®-based assay). Using this methodology, the association rate constant (ka) and the dissociation rate constant (kd) can be measured. The equilibrium dissociation constant (KD in M) can then be calculated from the ratio of the kinetic rate constants (kd / ka). In some embodiments, affinity is determined by a kinetic method, such as a Kinetic Exclusion Assay (KinExA) as described in Rathanaswami et al. Analytical Biochemistry,Vol. 373:52-60, 2008. Using a KinExA assay, the equilibrium dissociation constant (KD in M) and the association rate constant (ka in M'V1) can be measured. The dissociation rate constant (kd) can be calculated from these values (KD X ka). In other embodiments, affinity is determined by a bio-layer interferometry method, such as that described in Kumaraswamy et al., Methods Mol. Biol., Vol. 1278:165-82, 2015 and employed in Octet® systems (Pall ForteBio). The kinetic (ka and kd) and affinity (KD) constants can be calculated in real-time using the bio-layer interferometry method. In some embodiments, the antigen binding proteins described herein exhibit desirable characteristics such as binding avidity as measured by kd (dissociation rate constant) for human GIP of about 10'2, 10'3, 10’4, 10'5, 10'6or lower (lower values indicating higher binding avidity), and / or binding affinity as measured by KD (equilibrium dissociation constant) for human GIP of about 10-8, 109, 1010, 10-11M or lower (lower values indicating higher binding affinity). In certain embodiments, the antibodies or antibodies of the disclosure specifically bind to human GIP with a KD from about 1 pM to about 100 nM as measured by biolayer interferometry at 25° C. For instance, in some embodiments, the antibodies or antibodies of the disclosure specifically bind to human GIP with a KD less than 100 nM as measured by bio-layer interferometry at 25° C. In other embodiments, the antibodies or antibodies of the disclosure specifically bind to human GIP with a KD less than 50 nM as measured by bio-layer interferometry at 25° C embodiments, the antibodies or antibodies of the disclosure specifically bind to human GIP with a KD less than 25 nM as measured by bio-layer interferometry at 25° C. In one particular embodiment, the antibodies or antibodies of the disclosure specifically bind to human GIP with a KD less than 10 nM as measured by bio-layer interferometry at 25° C. In another particular embodiment, the antibodies or antibodies of the disclosure specifically bind to human GIP with a KD less than 5 nM as measured by bio-layer interferometry at 25° C. In another particular embodiment, the antibodies or antibodies of the disclosure specifically bind to human GIP and with a KD less than 1 nM as measured by bio-layer interferometry at 25° C.

[0051] As used herein, the term “variable region” or “variable domain” is used interchangeably and are common in the art. The variable region typically refers to a portion of an antibody, generally, a portion of a light or heavy chain, typically about the amino-terminal 110 to 120 amino acids in the mature heavy chain and about 90 to 115 amino acids in the mature light chain, which differ extensively in sequence among antibodies and are used in the binding and specificity of a particular antibody for its particular antigen. The variability in sequence is concentrated in those regions called CDRs while the more highly conserved regions in the variable domain are called framework regions (FR). Without wishing to be bound by anyparticular mechanism or theory, it is believed that the CDRs of the light and heavy chains are primarily responsible for the interaction and specificity of the antibody with antigen. In certain embodiments, the variable region is a human variable region. In certain embodiments, the variable region comprises rodent or murine CDRs and human framework regions (FRs). In particular embodiments, the variable region is a primate (e.g., non-human primate) variable region. In certain embodiments, the variable region comprises rodent or murine CDRs and primate (e.g., non-human primate) framework regions (FRs).

[0052] The term “Fc region,” “fragment crystallizable region,” “Fc fragment,” or “Fc domain” herein is used to define a C-terminal region of an immunoglobulin heavy chain, including native- sequence Fc regions and variant Fc regions. Although the boundaries of the Fc region of an immunoglobulin heavy chain might vary, the human IgG heavy-chain Fc region is usually defined to stretch from an amino acid residue at position Cys226, or from Pro230, to the carboxyl-terminus thereof. The C-terminal lysine (residue 447 according to the EU numbering system) of the Fc region may be removed, for example, during production or purification of the antibody or Fc- fusion protein, or by recombinantly engineering the nucleic acid encoding a heavy chain of the antibody or Fc-fusion protein. Accordingly, a composition of intact antibodies may comprise antibody populations with all K447 residues removed, antibody populations with no K447 residues removed, and antibody populations having a mixture of antibodies with and without the K447 residue. Suitable native-sequence Fc regions for use in the antibodies described herein include human lgG1 , lgG2 (lgG2A, lgG2B), lgG3 and lgG4. The term IgG “isotype” or “subclass” as used herein is meant any of the subclasses of immunoglobulins defined by the chemical and antigenic characteristics of their constant regions. There are five major classes of immunoglobulins: IgA, IgD, IgE, IgG, and IgM, and several of these may be further divided into subclasses (isotypes), e.g., lgG1 , lgG2, lgG3, lgG4, lgA1 , and lgA2. The heavy chain constant domains that correspond to the different classes of immunoglobulins are called a, y, E, y, and p, respectively. The subunit structures and three- dimensional configurations of different classes of immunoglobulins are well known and described generally in, for example, Abbas et al. Cellular and Mol. Immunology, 4th ed. (W.B. Saunders, Co., 2000).

[0053] “Fc receptor” or “FcR” describes a receptor that binds the Fc region of an antibody or Fc-fusion protein. The preferred FcR is a native sequence human FcR. Moreover, a preferred FcR is one which binds an IgG antibody (a gamma receptor) and includes receptors of the FcyRI, FcyRII, and FcyRIII subclasses, including allelic variants and alternatively spliced forms of these receptors, FcyRII receptors include FcyRIIA (an “activating receptor”) andFcyRIIB (an “inhibiting receptor”), which have similar amino acid sequences that differ primarily in the cytoplasmic domains thereof. Activating receptor FcyRIIA contains an immunoreceptor tyrosine- based activation motif (ITAM) in its cytoplasmic domain. Inhibiting receptor FcyRIIB contains an immunoreceptor tyrosine-based inhibition motif (ITIM) in its cytoplasmic domain. FcRs are reviewed in Ravetch and Kinet, Annu. Rev. Immunol.9: 457-92 (1991); Capel et al., Immunomethods 4: 25-34 (1994); and de Haas et al., J. Lab. Clin. Med. 126: 330-41 (1995). Other FcRs, including those to be identified in the future, are encompassed by the term “FcR” herein.

[0054] The antibodies of the disclosure have reduced antibody effector functions. The term “Antibody effector functions” refer to those biological activities attributable to the Fc region (a native sequence Fc region or amino acid sequence variant Fc region) of an antibody and vary with the antibody isotype. Examples of antibody effector functions include: C1q binding and complement dependent cytotoxicity; Fc receptor binding; antibody-dependent cell-mediated cytotoxicity (ADCC); phagocytosis; down regulation of cell surface receptors (e.g., B cell receptors); and B cell activation. “Reduced or minimized” antibody effector function means that which is reduced by at least 50% (alternatively 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99%) from the wild type or unmodified antibody. The determination of antibody effector function is readily determinable and measurable by one of ordinary skill in the art. In a preferred embodiment, the antibody effector functions of complement binding, complement dependent cytotoxicity and antibody dependent cytotoxicity are affected. In some embodiments, effector function is eliminated through a mutation in the constant region that eliminated glycosylation, e.g., “effectorless mutation.” Alternatively, effector function can be reduced or eliminated through production techniques, such as expression in host cells that do not glycosylate (e.g., E. coli.) or in which result in an altered glycosylation pattern that is ineffective or less effective at promoting effector function (e.g., Shinkawa et al., J. Biol. Chem. 278(5): 3466-3473 (2003). “Antibody-dependent cell-mediated cytotoxicity” or ADCC refers to a form of cytotoxicity in which secreted Ig bound onto Fc receptors (FcRs) present on certain cytotoxic cells (e.g., natural killer (NK) cells, neutrophils and macrophages) enable these cytotoxic effector cells to bind specifically to an antigen-bearing target cell and subsequently kill the target cell with cytotoxins. The antibodies “harm” the cytotoxic cells and are required for killing of the target cell by this mechanism. The primary cells for mediating ADCC, NK cells, express FcyRIII only, whereas monocytes express FcyRI, FcyRII, and FcyRIII. Fc expression on hematopoietic cells is summarized in Table 2 on page 464 of Ravetch and Kinet, Annu. Rev.Immunol.9: 457-92 (1991). To assess ADCC activity of a molecule of interest, an in vitro ADCC assay, such as that described in U.S. Pat. No. 5,500,362 or 5,821 ,337 may be performed. Useful effector cells for such assays include peripheral blood mononuclear cells (PBMC) and natural killer (NK) cells. Alternatively, or additionally, ADCC activity of the molecule of interest may be assessed in vivo, e.g., in an animal model such as that disclosed in Clynes et al., PNAS USA 95:652-656 (1998). “Complement dependent cytotoxicity” or “CDC” refers to the lysis of a target cell in the presence of complement. Activation of the classical complement pathway is initiated by the binding of the first component of the complement system (C1q) to antibodies (of the appropriate subclass) which are bound to their cognate antigen. To assess complement activation, a CDC assay, e.g., as described in Gazzano-Santoro et al., J. Immunol. Methods 202: 163 (1996), may be performed. Antibody variants with altered Fc region amino acid sequences and increased or decreased C1q binding capability are described in U.S. Pat. No. 6,194,551 B1 and WO99 / 51642. The contents of those patent publications are specifically incorporated herein by reference. See, also, Idusogie et al. J. Immunol.164: 4178-4184 (2000).

[0055] As used herein, an antigen binding molecule, an antibody, or an antigen binding molecule thereof, or a antibody of the disclosure “cross-competes” or ’’competes” with a reference antibody or an antigen binding molecule thereof or antibody of the disclosure if the interaction between an antigen and the first binding molecule, an antibody, or an antigen binding molecule or antibody of the disclosure blocks, limits, inhibits, or otherwise reduces the ability of the reference binding molecule, reference antibody, an antigen binding molecule thereof to interact with GIP. Cross competition may be complete, e.g., binding of the binding molecule to the antigen completely blocks the ability of the reference binding molecule to bind the antigen, or it may be partial, e.g., binding of the binding molecule to the antigen reduces the ability of the reference binding molecule to bind the antigen. In certain embodiments, an antigen binding molecule that cross-competes with a reference antigen binding molecule binds the same or an overlapping epitope as the reference antigen binding molecule. In other embodiments, the antigen binding molecule that cross-competes with a reference antigen binding molecule binds a different epitope as the reference antigen binding molecule.

[0056] Numerous types of competitive binding assays may be used to determine if one antigen binding molecule or polypeptide competes with another, for example: solid phase direct or indirect radioimmunoassay (RIA); solid phase direct or indirect enzyme immunoassay (EIA); sandwich competition assay (Stahli et al., 1983, Methods in Enzymology 9:242-253); solid phase direct biotin-avidin EIA (Kirkland et al., 1986, J. Immunol. 137:3614-3619); solid phasedirect labeled assay, solid phase direct labeled sandwich assay (Harlow and Lane, 1988, Antibodies, A Laboratory Manual, Cold Spring Harbor Press); solid phase direct label RIA using 1-125 label (Morel et al., 1988, Molec. Immunol. 25:7-15), solid phase direct biotin-avidin EIA (Cheung, et al., 1990, Virology 176:546-552), and direct labeled RIA (Moldenhauer et al., 1990, Scand. J. Immunol. 32:77-82). These methods are also examples of methods for measuring the affinity of the peptides of the disclosure for the antibodies or coronaviral S protein of the disclosure.

[0057] An “antigen” refers to any molecule that provokes an immune response or is capable of being bound by an antibody or an antigen binding molecule (e.g., a TCR). The immune response may involve either antibody production, or the activation of specific immunologically-competent cells, or both. A person of skill in the art would readily understand that any macromolecule, including virtually all proteins or peptides, could serve as an antigen. An antigen may be endogenously expressed, i.e. expressed by genomic DNA, or may be recombinantly expressed. An antigen may be specific to a certain tissue, such as a cancer cell, or it may be broadly expressed. In addition, fragments of larger molecules may act as antigens. In this disclosure, the antigen is GIP.

[0058] The terms “cell”, “cell culture”, “cell line” refer not only to the particular subject cell, cell culture, or cell line but also to the progeny or potential progeny of such a cell, cell culture, or cell line, without regard to the number of transfers or passages in culture. It should be understood that not all progeny are exactly identical to the parental cell. This is because certain modifications may occur in succeeding generations due to either mutation ( e.g ., deliberate or inadvertent mutations) or environmental influences (e.g., methylation or other epigenetic modifications), such that progeny may not, in fact, be identical to the parent cell, but are still included within the scope of the term as used herein, so long as the progeny retain the same functionality as that of the originally cell, cell culture, or cell line.

[0059] The term “ EC50” or “ EC50” refers to the half maximal effective concentration, which includes the concentration of an antibody which induces a response halfway between the baseline and maximum after a specified exposure time. The EC50 essentially represents the concentration of an antibody molecule where 50% of its maximal effect is observed.

[0060] The disclosure provides a GIP antagonist in the form of humanized antibody against GIP that has been optimized by neutralization of Fc-mediated inflammatory side effects through mutations on the Fc region of the antibody. One activity of the Fc domain of an antibodyis to bind complement proteins which can assist in lysing the target antigen, for example, a cellular pathogen. Another activity of the Fc region is to bind to Fc receptors (FcR) on the surface of immune cells, or so-called effector cells, which have the ability to trigger other immune effects. These immune effects include, for example, release of immune activators, regulation of antibody production, endocytosis, phagocytosis, and cell killing. In some clinical applications these responses are crucial for the efficacy of the antibody while in other cases they provoke unwanted side effects. One example of an effector-mediated side effect is the release of inflammatory cytokines causing an acute fever reaction. Another example is the long term deletion of antigen-bearing cells. In antibody-dependent cellular cytotoxicity (ADCC), an Fey receptor (FcyR or FCGR) on the surface of an immune effector cell binds to the Fc region of an antibody, which specifically binds to a target cell. Cells that can mediate ADCC are nonspecific cytotoxic cells such as natural killer cells, macrophages, monocytes, and eosinophils. When FCGR binds to the antibody, the FCGR ITAM is phosphorylated, which triggers the activation of effector cells and the secretion of various substances (lyase, perforin, granzyme, TNF) that mediate target cell destruction. In complement-dependent cytotoxicity (CDC), C1q binds to antibodies, which triggers the complement cascade, resulting in the formation of a membrane attack complex (MAC) (C5b to C9) on the surface of target cells, and further resulting in a classical pathway of complement activation. Antibody-dependent cellular phagocytosis (ADCP) is a key mechanism of action for many antibody therapies. It is defined as a highly regulated process in which an antibody eliminates binding target and initiates phagocytosis by linking its Fc domain to a specific receptor on the phagocytic cell. Unlike ADCC, ADCP can mediate monocytes, macrophages, neutrophils and dendritic cells via FcyRlla, FcyRI and FcyRllla, where FcyRlla (CD32a) on macrophages represents the major pathway.EXEMPLARY Fc VARIANTS OF THE DISCLOSURE

[0061] The present disclosure provides, among other things, Fc variants that have significantly reduced ADCC, ADCP and CDC function. These variants are referred to herein as Fc null variants. As described herein, the present disclosure is, in part, based on identification of novel combinations of mutations that abolish binding to all FcyRI, FcyRlla, FcyRllb, FcyRllla, FcyRlllb, and C1q.

[0062] The Fc variants of the present disclosure may find use in a variety of Fc polypeptides. An Fc polypeptide that comprises an Fc variant of the present disclosure is herein referred to as an “Fc polypeptide of the present disclosure”. Fc polypeptides of the present disclosure include polypeptides that comprise the Fc variants of the present disclosure in the context of a larger polypeptide, such as an antibody or Fc fusion. That is, Fc polypeptides of the present disclosure include antibodies and Fc fusions that comprise Fc variants of the present disclosure. By “antibody of the present disclosure” as used herein is meant an antibody that comprises an Fc variant of the present disclosure. By “Fc fusion of the present disclosure” as used herein refers to an Fc fusion that comprises an Fc variant of the present disclosure. Fc polypeptides of the present disclosure also include polypeptides that comprise little or no additional polypeptide sequence other than the Fc region, referred to as an isolated Fc. Fc polypeptides of the present disclosure also include fragments of the Fc region. As described below, any of the aforementioned Fc polypeptides of the present disclosure may be fused to one or more fusion partners or conjugate partners to provide desired functional properties.

[0063] In one embodiment, the Fc polypeptide of the present disclosure in an antibody. In one embodiment, the antibody has a human IgG constant region wherein the Fc domain comprises at least one amino acid substitution that reduces binding affinity to an Fc receptor and / or effector function. In some embodiments, an Fc variant comprises one or more, two or more, three or more, four or more, or all five amino acid substitutions selected from L234A, L235A, G237A, K322A, and P329G, wherein the residues are numbered according to the EU index. In some embodiments, an Fc variant comprises the amino acid substitution L234A. In some embodiments, an Fc variant comprises the amino acid substitution L235A. In some embodiments, an Fc variant comprises the amino acid substitution G237A. In some embodiments, an Fc variant comprises the amino acid substitution K322A. In some embodiments, an Fc variant comprises the amino acid substitution P329G. In some embodimenets, the Fc variant comprises two or more amino acid substitutions selected from L234A and at least one of L235A, G237A, K322A, and P329G. In some embodiments, the Fc variant comprises two or more amino acid substitutions selected from L235A and at least one of L234A, G237A, K322A, and P329G. In some embodiments, the Fc variant comprises two or more amino acid substitutions selected from G237A and at least one of L234A, L235A, K322A, and P329G. In some embodiments, the Fc variant comprises two or more amino acid substitutions selected from K322A and at least one L235A, G237A, K322A, and P329G. Insome embodiments, the Fc variant comprises two or more amino acid substitutions selected from P329G and at least one L234A, L235A, G237A, and K322A.

[0064] In one embodiment, the disclosure provides Fc null versions of the humanized anti-GIP antibody known as NM-136, which is an IgG 1 antibody. The Fc null antibodies may carry one or more of the amino acid substitutions selected from L234A, L235A, G237A, K322A, and P329G, wherein the residues are numbered according to the EU index.

[0065] The NM-136 antibody (INC-118) comprises the following heavy chain (HC) and light chain (LC) complementarity determining regions (CDRs), according to Kabat:

[0066] HC-CDR1 : DYYLH (SEQ ID NO: 1)

[0067] HC-CDR2: WIDPENGDTEYAPKFQG (SEQ ID NO: 2)

[0068] HC-CDR3: YGIYFMDY (SEQ ID NO: 3)

[0069] LC-CDR1 : RASSSISSNSLH (SEQ ID NO: 4)

[0070] LC-CDR2: RTSNLQS (SEQ ID NO: 5)

[0071] LC-CDR3: QQGSSFPRMLT (SEQ ID NO: 6)

[0072] In one embodiment, the NM-136 antibody comprises the heavy chain variable region of SEQ ID NO: 7:

[0073] QVQLVQSGAEVKKPGATVKISCKASGFNIRDYYLHWVRQAPGKGLEWMGWID PENGDTEYAPKFQGRVTITADTSTNTAYMELSSLRSEDTAVYYCNVYGIYFMDYWGQGTMVTV SS.

[0074] In one embodiment, the NM-136 antibody comprises the light chain variable region of SEQ ID NO: 8:

[0075] DIQLTQSPSSVSASVGDRVTITCRASSSISSNSLHWYQQKPGKAPKLLIYRTSNL QSGVPSRFSGSGSGTDYTLTISSLQPEDFATYYCQQGSSFPRMLTFGQGTKLEIK

[0076] In one embodiment, the NM-136 Fc variant comprises the heavy chain constant region of SEQ ID NO: 9:

[0077] QVQLVQSGAEVKKPGATVKISCKASGFNIRDYYLHWVRQAPGKGLEWMGWID PENGDTEYAPKFQGRVTITADTSTNTAYMELSSLRSEDTAVYYCNVYGIYFMDYWGQGTMVTV SSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSG LYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFL FPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSV LTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLV KGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEA LHNHYTQKSLSLSPGK.

[0078] In one embodiment, the NM-136 Fc variant comprises the light chain constant region of SEQ ID NO: 10:

[0079] DIQLTQSPSSVSASVGDRVTITCRASSSISSNSLHWYQQKPGKAPKLLIYRTSNL QSGVPSRFSGSGSGTDYTLTISSLQPEDFATYYCQQGSSFPRMLTFGQGTKLEIKRTVAAPSV FIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSST LTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC.

[0080] In one embodiment, the NM-136 Fc variant comprises the heavy chain constant region of SEQ ID NO: 11 containing the following mutations: L234A, L235A, G237A, K322A and P329G:

[0081] QVQLVQSGAEVKKPGATVKISCKASGFNIRDYYLHWVRQAPGKGLEWMGWID PENGDTEYAPKFQGRVTITADTSTNTAYMELSSLRSEDTAVYYCNVYGIYFMDYWGQGTMVTV SSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGAPSVFL FPPKPKDTLMISRTPEVTCWVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSV LTVLHQDWLNGKEYKCAVSNKALGAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLV KGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEA LHNHYTQKSLSLSPGK

[0082] In one embodiment, the NM-136 Fc variant comprises the heavy chain constant region of SEQ ID NO: 12 containing the following mutations: L234A, L235A, G237A, K322A and P329G, and M252Y, S254T, T256E:QVQLVQSGAEVKKPGATVKISCKASGFNIRDYYLHWVRQAPGKGLEWMGWIDPENGDTEYAP KFQGRVTITADTSTNTAYMELSSLRSEDTAVYYCNVYGIYFMDYWGQGTMVTVSSASTKGPSV FPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTV PSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLY ITREPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWL NGKEYKCAVSNKALGAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIA VEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQK SLSLSPGK

[0083] In one embodiment, the NM-136 Fc variant comprises the heavy chain constant region of SEQ ID NO: 13 containing the following mutations: L234A, L235A, G237A, K322A and P329G, and M428L, and N434S:

[0084] QVQLVQSGAEVKKPGATVKISCKASGFNIRDYYLHWVRQAPGKGLEWMGWID PENGDTEYAPKFQGRVTITADTSTNTAYMELSSLRSEDTAVYYCNVYGIYFMDYWGQGTMVTVSSASTKGPSVFPLAPSSKSTSGGTALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGL YSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGAPSVFLF PPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRWSVL TVLHQDWLNGKEYKCAVSNKALGAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVK GFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEAL HSHYTQKSLSLSPGK

[0085] In one embodiment, the disclosure provides a murinized lncre-002 with engineered mu Fc. The antibody comprises the heavy chain of SEQ ID NO: 14 and the light chain of SEQ ID NO: 15, as follows:SEQ ID: NO 14:QVQLQQSGPELVKPGASVKISCKASGFNIRDYYLHWVRQAPGQGLEWMGWIDPENGDTEYA PKFQGRVTLTADTSSSTAYMQLSSLTSEDSAVYYCNVYGIYFMDYWGQGTSVTVSSAKTTAPS VYPLAPVCGDTTGSSVTLGCLVKGYFPEPVTLTWNSGSLSSGVHTFPAVLQSDLYTLSSSVTV TSSTWPSQSITCNVAHPASSTKVDKKIEPRGPTIKPCPPCKCPAPNAAGAPSVFIFPPKIKDVLM ISLSPIVTCVVVDVSEDDPDVQISWFVNNVEVHTAQTQTHREDYNSTLRVVSALPIQHQDWMS GKEFKCAVNNKDLGAPIERTISKPKGSVRAPQVYVLPPPEEEMTKKQVTLTCMVTDFMPEDIYV EWTNNGKTELNYKNTEPVLDSDGSYFMYSKLRVEKKNWVERNSYSCSVVHEGLHNHHTTKSF SRTPGKSEQ ID: 15:DIQMTQTTSSLSASLGDRVTISCRASSSISSNSLHWYQQKPDGTVKLLIYRTSNLQSGVPSRFS GSGSGTDYSLTISNLEPEDIATYYCQQGSSFPRMLTFGSGTKLEIKRADAAPTVSIFPPSSEQLT SGGASVVCFLNNFYPKDINVKWKIDGSERQNGVLNSWTDQDSKDSTYSMSSTLTLTKDEYER HNSYTCEATHKTSTSPIVKSFNRNECEXEMPLARY APPLICATIONS

[0086] In one aspect, the present disclosure provides a method of treating a disease or disorder, said method comprising administering a therapeutically effective amount of an antibody of the disclosure to a subject in need thereof. In one embodiment, the antibodies of the disclosure have wide-spread uses in the treatment of obesity and obesity-associated disorders.

[0087] Obesity is a medical condition in which excess body fat has accumulated to the extent that it may have an adverse effect on health and life expectancy and due to its increasing prevalence in adults and children it has become one of the leading preventable causes of deathin modern world. It increases the likelihood of various other diseases, including heart disease, type 2 diabetes, obstructive sleep apnoea, certain types of cancer, as well as osteoarthritis, and it is most commonly caused by a combination of excess food intake, reduced energy expenditure, as well as genetic susceptibility.

[0088] In terms of a human (adult) subject, obesity can be defined as a body mass index (BMI) greater than or equal to 30 kg / m2(BMI>30 kg / m2). The BMI is a simple index of weight- for-height that is commonly used to classify overweight and obesity in adults. It is defined as a person's weight in kilograms divided by the square of his / her height in meters (kg / m2).

[0089] The term “overweight” refers to a medical condition in which the amount of body fat is higher than is optimally healthy. In terms of a human (adult) subject, overweight can be defined as a body mass index (BMI) greater than or equal to 25 kg / m2(e.g., 25 kg / m2<BMI<30 kg / m2)

[0090] Obesity-associated disorders include several chronic diseases, such as tumors, hypertension, type 2 diabetes mellitus (T2DM), cerebrovascular diseases, and chronic kidney disease. Also, adipose tissue increases in pharyngeal soft tissue, causing blocked airways during sleep and triggering obstructive sleep apnea. Excess adiposity also determines both osteoarthritis, due to increased mechanical loading on the joints, and gastroesophageal reflux disease, due to an increase in intra-abdominal pressure. In obesity, an increase of macrophages and other immune cells in adipose tissue has been described, resulting in an increase in pro-inflammatory cytokines, promoting insulin resistance. Furthermore, insulin secretion increases linearly with the BMI, and insulin resistance favors dyslipidemia and T2DM. This obesity-induced chronic inflammation furthers cancer progression due to its promotion of a permissive microenvironment for neoplastic transformation. Furthermore, liposomes augment in hepatocytes that evolve in non-alcoholic fatty liver disease, steatohepatitis, and cirrhosis.Another consequence of obesity is chronic overactivity of the sympathetic nervous system, which, together with the previously described consequences of chronic obesity, induces hypertension and increases the risk of heart disease, stroke, and chronic kidney dysfunction. Accordingly, the disclosure provides methods for treatment of obesity and obesity-related disorders.

[0091] In one embodiment, the disclosure relates to the treatment or amelioration of a metabolic disorder, such as type 2 diabetes, elevated glucose levels, elevated insulin levels, obesity, nonalcoholic fatty liver disease, cardiovascular diseases, or diabetic nephropathy.

[0092] In one embodiment, the metabolic disorder is a disorder of glucose metabolism. In another embodiment, the glucose metabolism disorder comprises hyperglycemia and administering the Fc-null antibody reduces plasma glucose. In another embodiment, the glucose metabolism disorder comprises hyperinsulinemia and administering the Fc-null antibody reduces plasma insulin. In another embodiment, the glucose metabolism disorder comprises glucose intolerance and administering the Fc-null antibody reduces increases glucose tolerance. In another embodiment, the glucose metabolism disorder comprises insulin resistance and administering the Fc-null antibody reduces insulin resistance. In another embodiment, the glucose metabolism disorder comprises diabetes mellitus. In another embodiment, the subject is obese. In another embodiment, administering the composition reduces body weight in an obese subject. In another embodiment, administering the composition reduces body weight gain in an obese subject. In another embodiment, administering the composition reduces fat mass in an obese subject. In another embodiment, the glucose metabolism disorder comprises insulin resistance and administering the composition reduces insulin resistance in an obese subject. In another embodiment, administering the composition reduces liver steatosis in an obese subject having increased liver steatosis. In another embodiment, administering the composition reduces liver fat content in an obese subject having increased liver fat content. In some embodiments, the antibodies of the disclosure may stimulate significant weight loss by reducing gastric emptying, stimulating satiety, and decreasing food intake by acting on peripheral and central receptors in the gut and brain. In some embodiments, the antibodies of the disclosure may reduce blood pressure, improve renal function, reduce chronic inflammation, reduce lipoprotein, reduce chylomicron, increase postprandial triglycerides, increase very-low-density lipoprotein cholesterol (VLDL-C), and increase free fatty acids.

[0093] MAINTENANCE OF WEIGHT LOSS

[0094] In one embodiment, the disclosure relates to a method of maintaining weight loss in a subject that has lost weight, comprising administering a therapeutically effective amount of an antibody or a composition of the disclosure to the subject.

[0095] In one embodiment, the disclosure relates to a method of preventing weight regain in a subject that has lost weight, comprising administering a therapeutically effective amount of an antibody or a composition of the disclosure to the subject.

[0096] In one embodiment, the disclosure relates to a method of preventing the “yo-yo effect” in a subject in need thereof related to constant fluctuation in weight related to losingweight then regaining the weight (or more weight) after stopping a diet, comprising administering a therapeutically effective amount of an antibody or a composition of the disclosure to the subject.

[0097] In the context of this application, the "yo-yo effect" is a medically recognized phenomenon where people lose weight upon dieting, life-style changes, and / or treatment with one or more drugs that cause or associated with weight loss (many of which are described herein) and then regain it after stopping their diet and / or treatment. Many experts believe that weight swings related to these fluctuations can reduce health benefits related to weight loss.COMBINATION TREATMENTS

[0098] In some embodiments, the antibodies are used in combination with one or more additional therapeutic agents. In some embodiments, the disclosure features a method wherein the subject is treated with one or more additional therapeutic agents administered concurrently with, prior to, and / or subsequent to treatment with an effective amount of the compound, pharmaceutically acceptable salt or pharmaceutical compositions of the disclosure. The additional therapeutic agent may be in its own separate composition or in a composition that comprises both the antibody and one or more additional therapeutic agents.

[0099] In some embodiments, the antibodies of the disclosure are used in combination with behavioral therapy (diet and lifestyle modifications), other drugs, and bariatric surgery. In one embodiment, antibody and other drug therapies may be considered for patients with a BMI of >30 kg / m2 and a BMI of >27 kg / m2 with weight-related comorbidities. In one embodiment, the disclosure provides treatments that stimulate satiety, reduce hunger, and / or reduce fat absorption or catabolism.

[0100] In one embodiment, the antibodies of the disclosure may be combined with a GLP-1 receptor agonist. In one embodiment, the antibodies of the disclosure may be combined with one or more GLP-1-RAs, such as dulaglutide and semaglutide. In one embodiment, the antibodies of the disclosure may be combined with liraglutide. The antibodies of the disclosure may also be used in combination with any other drugs approved for the treatment of obesity, such as orlistat, setmelanotide, metreleptin, phentermine, topiramate, naltrexone, bupropion, liraglutide, semaglutide, setmelanotide, tirzepatide, and the like.

[0101] In some embodiments, the antibodies of the disclosure may be administered in combination with an anti-inflammatory agent. Exemplary anti-inflammatory agents or drugs may include, but are not limited to, steroids and glucocorticoids (including betamethasone,budesonide, dexamethasone, hydrocortisone acetate, hydrocortisone, hydrocortisone, methylprednisolone, prednisolone, prednisone, triamcinolone), nonsteroidal anti-inflammatory drugs (NSAIDS) including aspirin, ibuprofen, naproxen, methotrexate, sulfasalazine, leflunomide, anti-TNF medications, cyclophosphamide and mycophenolate. Exemplary NSAIDs include ibuprofen, naproxen, naproxen sodium, Cox-2 inhibitors, and sialylates. Exemplary analgesics include acetaminophen, oxycodone, tramadol of proporxyphene hydrochloride. Exemplary glucocorticoids include cortisone, dexamethasone, hydrocortisone, methylprednisolone, prednisolone, or prednisone. Exemplary biological response modifiers include molecules directed against cell surface markers (e.g., CD4, CD5, etc.), cytokine inhibitors, such as the TNF antagonists, (e.g., etanercept (ENBREL®), adalimumab (HUMIRA®) and infliximab (REMICADE®), chemokine inhibitors and adhesion molecule inhibitors. The biological response modifiers include monoclonal antibodies as well as recombinant forms of molecules. Exemplary DMARDs include azathioprine, cyclophosphamide, cyclosporine, methotrexate, penicillamine, leflunomide, sulfasalazine, hydroxychloroquine, Gold (oral (auranofin) and intramuscular), and minocycline.

[0102] COMPOSITIONS

[0103] The disclosure provides that the antibodies / compounds of the disclosure may be prepared through any process (e.g., as recombinant proteins) and provided alone or in compositions. In some embodiments, the compositions are pharmaceutically acceptable compositions. The terms “pharmaceutical composition” and ’’pharmaceutically acceptable composition” are used interchangeably and indicate a mixture containing ingredients that are compatible when mixed and which may be administered. A pharmaceutical composition may include one or more medicinal drugs. Additionally, the pharmaceutical composition may include carriers, buffers, acidifying agents, alkalizing agents, solvents, adjuvants, tonicity adjusters, emollients, expanders, preservatives, physical and chemical stabilizers e.g. surfactants, antioxidants and other components, whether these are considered active or inactive ingredients. Guidance for the skilled in preparing pharmaceutical compositions may be found, for example, in Remington: The Science and Practice of Pharmacy, (20th ed.) ed. A. R. GennaroA. R., 2000, Lippencott Williams & Wilkins and in R. C. Rowe et al. (Ed), Handbook of Pharmaceutical Excipients, PhP, 9th edition (2020).

[0104] In some embodiments, the pharmaceutically acceptable compositions of the disclosure additionally comprise a pharmaceutically acceptable carrier, adjuvant, or vehicle, which, as used herein, includes any and all solvents, diluents, or other liquid vehicle, dispersion or suspension aids, surface active agents, isotonic agents, thickening or emulsifying agents, preservatives, solid binders, lubricants and the like, as suited to the particular dosage form desired. Remington’s Pharmaceutical Sciences, 22nd Edition (Sep. 15, 2022) discloses various carriers used in formulating pharmaceutically acceptable compositions and known techniques for the preparation thereof. Except insofar as any conventional carrier medium is incompatible with the antibodies / compounds of the disclosure, such as by producing any undesirable biological effect or otherwise interacting in a deleterious manner with any other component(s) of the pharmaceutically acceptable composition, its use is contemplated to be within the scope of this disclosure. Some examples of materials which may serve as pharmaceutically acceptable carriers include, but are not limited to, ion exchangers, alumina, aluminum stearate, lecithin, serum proteins, such as human serum albumin, buffer substances such as phosphates, glycine, sorbic acid, or 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 silica, magnesium trisilicate, polyvinyl pyrrolidone, polyacrylates, waxes, polyethylene-polyoxypropylene-block polymers, wool fat, sugars such as lactose, glucose and sucrose; starches such as corn starch and potato starch; cellulose and its derivatives such as sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate; powdered tragacanth; malt; gelatin; talc; excipients such as cocoa butter and suppository waxes; oils such as peanut oil, cottonseed oil; safflower oil; sesame oil; olive oil; corn oil and soybean oil; glycols; such a propylene glycol or polyethylene glycol; esters such as ethyl oleate and ethyl laurate; agar; buffering agents such as magnesium hydroxide and aluminum hydroxide; alginic acid; pyrogen-free water; isotonic saline; Ringer’s solution; ethyl alcohol, and phosphate buffer solutions, as well as other non-toxic compatible lubricants such as sodium lauryl sulfate and magnesium stearate, as well as coloring agents, releasing agents, coating agents, sweetening, flavoring and perfuming agents, preservatives and antioxidants may also be present in the composition, according to the judgment of the formulator. In a specific embodiment and in this context, the term “pharmaceutically acceptable carrier or diluent” may mean approved by a regulatory agency of the Federal or a state government or listed in the U.S. Pharmacopeia or other generally recognized pharmacopeia for use in animals, and more particularly in humans.

[0105] In another embodiment, the disclosure features a pharmaceutical composition comprising an antibody / compoundof the disclosure, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier. In another embodiment, the disclosure features a pharmaceutical composition comprising a therapeutically effective amount of a compound, or a pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable carriers or vehicles.

[0106] The composition may include a solubilizing agent. The solubilizing agent may include a solvent or solvent system for Oxazolidone-derived compounds, and this solvent system, itself including one or more solvents, may form the bulk of the medium in which the Oxazolidone-derived compounds is dissolved. Regardless of the nature of the solubilizing agent and whether it includes one or more solvents, a sufficient quantity of the solubilizing agent is present to solubilize essentially all of the Oxazolidone-derived compounds. The solubilizing agent must be pharmaceutically acceptable when present in an amount needed to solubilize the Oxazolidone-derived compounds. For example, the solubilizing agent should not be toxic to nor cause excessive irritation of tissues lining the nasal cavity. In an embodiment, the solvent may be water, alcohol, or a combination thereof. In another embodiment, the solvent may be water.

[0107] The composition optionally further includes a receptivity agent. The term “receptivity agent” herein means an agent that, when included in a pharmaceutical composition administered to a subject, is capable of mitigating an undesirable response to the composition at or in proximity to the locus of administration in or on the subject. Specifically, when the locus of administration is intranasal, such undesirable responses that may be mitigated may include an involuntary or reflex response such as sneezing, excessive nasal drip or irritation of nasal tissues, and / or a cognitive response, such as to unpleasant taste or odor. A cognitive response may include a conscious or subconscious decision to reduce or end use of the composition, and may thus affect patient compliance. A receptivity agent may mitigate one or more such undesirable responses.

[0108] In some embodiments, the receptivity agent includes an organoleptic enhancing agent. Illustrative examples of organoleptic enhancing agents include natural and / or synthetic sweeteners, flavorants, aromatics, taste-masking compounds, or combinations thereof.

[0109] In some embodiments, an organoleptic enhancing agent included as a receptivity agent includes a sweetener. Illustrative sweeteners include saccharin, aspartame, neotame, cyclamates, glucose, fructose, sucrose, xylitol, tagatose, sucralose, maltitol, isomaltulose, hydrogenated isomaltulose, lactitol, sorbitol, mannitol, trehalose, maltodextrin, polydextrose,glycerin, erythritol, maltol, acesulfame, acesulfame potassium, alitame, neohesperidin dihydrochalcone, stevioside, thaumatin, sugars, or combinations thereof. In an embodiment, the receptivity agent includes an agent that may inhibit sneezing, i.e., an anti -sternutatory agent.

[0110] The pharmaceutical composition optionally further includes one or more pharmaceutically acceptable ingredients, for example, ingredients useful as carriers, preservatives, diluents, stabilizers, pH modulating agents, etc. According to an embodiment, the composition includes at least one preservative. Preservatives may have antimicrobial activity and / or may serve as antioxidants. Illustrative preservatives include but are not limited to butylated hydroxytoluene, butylated hydroxyanisole, or combinations thereof.

[0111] Where the composition is formulated in an aqueous medium, it may include one or more tonicity modulating agents, for example in an amount that renders the composition substantially isotonic. For example, a saline solution may form the basis of such a composition.

[0112] Active ingredients such as the compounds provided herein can be administered by controlled release means or by delivery devices that are well known to those of ordinary skill in the art. In some or any embodiments, but are not limited to, those described in U.S. Pat. Nos. 3,845,770; 3,916,899; 3,536,809; 3,598,123; 4,008,719; 5,674,533; 5,059,595; 5,591,767; 5,120,548; 5,073,543; 5,639,476; 5,354,556; 5,639,480; 5,733,566; 5,739,108; 5,891,474; 5,922,356; 5,972,891 ; 5,980,945; 5,993,855; 6,045,830; 6,087,324; 6,113,943; 6,197,350; 6,248,363; 6,264,970; 6,267,981; 6,376,461; 6,419,961 ; 6,589,548; 6,613,358; and 6,699,500; each of which is incorporated herein by reference in its entirety. Such dosage forms can be used to provide slow or controlled release of one or more active ingredients using, in some or any embodiments, hydropropylmethyl cellulose, other polymer matrices, gels, permeable membranes, osmotic systems, multilayer coatings, microparticles, liposomes, microspheres, or a combination thereof to provide the desired release profile in varying proportions. Suitable controlled release formulations known to those of ordinary skill in the art, including those described herein, can be readily selected for use with the active ingredients provided herein. Thus encompassed herein are unit dosage forms suitable for oral administration such as, but not limited to, tablets, capsules, gel caps, and caplets that are adapted for controlled release. All controlled release pharmaceutical products have a common goal of improving drug therapy over that achieved by their non-controlled counterparts. Ideally, the use of an optimally designed controlled release preparation in medical treatment is characterized by a minimum of drug substance being employed to cure or control the condition in a minimum amount of time. Advantages of controlled release formulations include extended activity of the drug, reduceddosage frequency, and increased subject compliance. In addition, controlled release formulations can be used to affect the time of onset of action or other characteristics, such as blood levels of the drug, and can thus affect the occurrence of side (e.g., adverse) effects. Most controlled release formulations are designed to initially release an amount of drug (active ingredient) that promptly produces the desired therapeutic effect, and gradually and continually release of other amounts of drug to maintain this level of therapeutic or prophylactic effect over an extended period of time. In order to maintain this constant level of drug in the body, the drug must be released from the dosage form at a rate that will replace the amount of drug being metabolized and excreted from the body. Controlled release of an active ingredient can be stimulated by various conditions including, but not limited to, pH, temperature, enzymes, water, or other physiological conditions or compounds.

[0113] In some embodiments, the composition is a composition (which may be for immediate or delayed / extended release). In some embodiments, the composition (which may be for immediate or delayed / extended release) can comprise any stabilizing amount of a cation (e.g., metal cation). Any suitable cation(s) can be included in the composition, for example, any suitable metal cation or organic cation. In some embodiments, the composition comprises a metal cation selected from calcium, potassium, magnesium, zinc, aluminum, iron, tin, manganese, chromium, cobalt, nickel, barium, sodium, or a combination or mixture thereof. In some embodiments, the composition comprises a metal cation selected from calcium, potassium, magnesium, zinc, aluminum, manganese, chromium, cobalt, nickel, barium, sodium, or a combination or mixture thereof. In some embodiments, the composition comprises a metal cation selected from aluminum, calcium, potassium, sodium, magnesium, manganese, zinc, or a combination or mixture thereof. In some embodiments, the composition comprises a metal cation selected from calcium, magnesium, manganese, zinc, or a combination or mixture thereof. In some embodiments, the composition comprises a divalent metal cation. In some embodiments, the composition comprises a divalent metal cation selected from AI3+, Ca2+, Mg2+, Zn2+, Mn2+, or a combination or mixture thereof. In some embodiments, the composition comprises Mg2+. In some embodiments, the composition comprises Ca2+. In some embodiments, the composition comprises Zn2+. In some embodiments, the composition comprises AI3+.

[0114] Moreover, the metal cation can be added to the composition in any suitable form, for example any pharmaceutically acceptable salt with any appropriate counterion. Suitable metal salts include, for example, calcium chloride, calcium carbonate, calcium acetate,magnesium chloride, magnesium acetate, zinc acetate, zinc chloride, or mixtures thereof. In some embodiments, the composition comprises calcium chloride, magnesium chloride, zinc acetate, or any combination or mixture thereof. In some embodiments, the composition comprises calcium chloride. In some embodiments, the composition comprises magnesium chloride. In some embodiments, the composition comprises zinc acetate. Suitable organic cations include, for example, ammonium hydroxide, D-arginine, L-arginine, t-butylamine, calcium acetate hydrate, calcium carbonate, calcium DL-malate, calcium hydroxide, choline, ethanolamine, ethylenediamine, glycine, L-histidine, L-lysine, magnesium hydroxide, N-methyl- D-glucamine, L-ornithine hydrochloride, potassium hydroxide, procaine hydrochloride, L-proline, pyridoxine, L-serine, sodium hydroxide, DL-tryptophan, tromethamine, L-tyrosine, L-valine, carnitine, taurine, creatine malate, arginine alpha ketoglutarate, ornithine alpha ketoglutarate, spermine acetate, spermidine chloride, or combinations or mixtures thereof. In some embodiments, the organic cation is selected from the group consisting of N-methyl D-glucamine, choline, arginine, lysine, procaine, tromethamine (TRIS), spermine, N-methyl-morpholine, glucosamine, N,N-bis(2-hydroxyethyl) glycine, diazabicycloundecene, creatine, arginine ethyl ester, amantadine, rimantadine, ornithine, taurine, and citrulline, or any combination or mixture thereof.

[0115] The composition (which may be for immediate or delayed / extended release) (e.g., delayed release tablet) can comprise any suitable concentration of filling agent. In some embodiments, for example, the composition comprises one or more filling agents in a concentration of 0.1-99% by weight, relative to the total weight of the composition. In some embodiments, for example, the composition comprises one or more filling agents in a concentration of 1-95 wt. % of filling agent(s), relative to the total weight of the composition. In some embodiments, for example, the composition comprises one or more filling agents in a concentration of 10-90 wt. % of filling agent(s), relative to the total weight of the composition. In some embodiments, for example, the composition comprises one or more filling agents in a concentration of 20-90 wt. % of filling agent(s), relative to the total weight of the composition. In some embodiments, for example, the composition comprises one or more filling agents in a concentration of 25-85 wt. % of filling agent(s), relative to the total weight of the composition. In some embodiments, for example, the composition comprises one or more filling agents in a concentration of 30-80 wt. % of filling agent(s), relative to the total weight of the composition. In some embodiments, for example, the composition comprises one or more filling agents in a concentration of 40-70 wt. % of filling agent(s), relative to the total weight of the composition. Insome embodiments, for example, the composition comprises one or more filling agents in a concentration of 10-60 wt. % of filling agent(s), relative to the total weight of the composition. In some embodiments, for example, the composition comprises one or more filling agents in a concentration of 20-50 wt. % of filling agent(s), relative to the total weight of the composition. In some embodiments, the composition comprises one or more filling agents in a concentration of at least 20 wt. %, for example, at least 40 wt. %, at least 60 wt. %, at least 70 wt. %, at least 80 wt. %, or at least 90 wt. %, relative to the total weight of the composition.

[0116] In some embodiments, the composition (which may be for immediate or delayed / extended release) (e.g., delayed release film) comprises one or more plasticizers. Suitable plasticizers include, but are not limited to, polyethylene glycol, propylene glycol, glycerin, glycerol, monoacetin, diacetin, triacetin, dimethyl phthalate, diethyl phthalate, dibutyl phthalate, dibutyl sebacate, triethyl titrate, tributyl citrate, triethyl citrate, triethyl acetyl citrate, castor oil, acetylated monoglycerides, sorbitol or combinations thereof. In exemplary embodiments, the concentration of the plasticizer in the formulation may be about 0 to about 30 wt. %, for example, about 1 to about 20 wt. %, about 0 to about 10 wt. %, about 1 to about 5 wt. %, or even 0 to about 4 wt. %.

[0117] In some embodiments, the composition (which may be for immediate or delayed / extended release) comprises a film forming agent, a water-soluble polymer, a pH sensitive polymer, biodegradable polymer, or combination thereof. Water soluble, pH sensitive, or biodegradable polymers that may be used in the orally dissolving formulations of the present disclosure include, but are not limited to, cellulose derivatives, synthetic polymers polyacrylates and natural gums. For example, the water soluble polymers used in the orally dissolving formulations of the present disclosure may include, but are not limited to, methyl cellulose, hydroxypropyl cellulose, hydroxypropylmethyl cellulose, ethyl cellulose, hydroxyethyl cellulose, hydroxypropyl cellulose, carboxymethyl cellulose, cellulose acetate phthalate, cellulose acetate butyrate, amylose, dextran, casein, pullulan, gelatin, pectin, agar, carrageenan, xanthan gum, tragacanth, guar gum, acacia gum, arabic gum, polyethylene glycol, polyethylene oxide, polyvinyl pyrrolidone, polyvinyl alcohol, cyclodextrin, carboxyvinyl polymers, sodium alginate, polyacrylic acid, methylmethacrylate or mixtures thereof. In exemplary embodiments, the concentration of the water-soluble polymer in the formulation may be about 20% to about 90% (by weight), preferably between about 40% to about 80% (by weight).

[0118] Suitable disintegrants include, for example, agar-agar, calcium carbonate, microcrystalline cellulose, croscarmellose sodium, crospovidone, povidone, polacrilinpotassium, sodium starch glycolate, potato or tapioca starch, other starches, pre-gelatinized starch, clays, other algins, other celluloses, gums, and mixtures thereof. In some embodiments, the disintegrant is crospovidone. In some embodiments, the disintegrant is croscarmellose sodium.

[0119] Suitable lubricants include, for example, calcium stearate, magnesium stearate, mineral oil, light mineral oil, glycerin, sorbitol, mannitol, polyethylene glycol, other glycols, stearic acid, sodium lauryl sulfate, talc, hydrogenated vegetable oil (e.g., peanut oil, cottonseed oil, sunflower oil, sesame oil, olive oil, corn oil and soybean oil), zinc stearate, ethyl oleate, ethyl laurate, agar, syloid silica gel (AEROSIL® 200, W.R. Grace Co., Baltimore, MD USA), a coagulated aerosol of synthetic silica (Evonik Degussa Co., Plano, TX USA), a pyrogenic silicon dioxide (CAB-O-SIL, Cabot Co., Boston, MA USA), and mixtures thereof.

[0120] Suitable anti-caking additives include, for example, calcium silicate, magnesium silicate, silicon dioxide, colloidal silicon dioxide, talc, glyceryl, and mixtures thereof.

[0121] Suitable anti-microbial additives that may be used, e.g., as a preservative for the a antibody / compound of the disclosure and / or (pharmaceutically acceptable) salt thereof compositions, include, for example, benzalkonium chloride, benzethonium chloride, benzoic acid, benzyl alcohol, butyl paraben, cetylpyridinium chloride, cresol, chlorobutanol, dehydroacetic acid, ethylparaben, methylparaben, phenol, phenylethyl alcohol, phenoxyethanol, phenylmercuric acetate, phenylmercuric nitrate, potassium sorbate, propylparaben, sodium benzoate, sodium dehydroacetate, sodium propionate, sorbic acid, thimersol, thymol, and mixtures thereof.

[0122] The composition may also comprise any suitable pharmaceutically acceptable carrier or medium. Suitable pharmaceutically acceptable carriers include, for example, any solvents, dispersants, pH buffering agents, coatings, absorption promoting agents, controlled release agents, and one or more inert excipients (e.g., filling agents, starches, polyols, granulating agents, microcrystalline cellulose, diluents, lubricants, binders, disintegrating agents), or the like. In addition, the compositions can contain any desired additional components, additives, and / or species, for example, surface active additives, dispersing additives, humectants, suspending agents, solubilizers, buffering agents, disintegrants, preservatives, colorants, flavorants, and the like. In some embodiments, the composition comprises one or more ion species that interact with the antibodies / compounds of the disclosure.

[0123] The composition can contain any stabilizing amount of a polymer. In some embodiments, the composition comprises between 1 and 25% by weight of a polymer, relative to the total weight of the composition. In some embodiments, the composition comprises between 1 and 10% by weight of a polymer, relative to the total weight of the composition. In some embodiments, the composition comprises between 2 and 4% by weight of a polymer, relative to the total weight of the composition. In some embodiments, the composition comprises between 0.01 and 5 wt. % of a polymer. In some embodiments, the composition comprises between 0.1 and 4 wt. % of a polymer. In some embodiments, the composition comprises about 0.71 wt. % of a polymer. In some embodiments, the composition comprises about 3.59 wt. % of a polymer.

[0124] In some embodiments, a antibody / compound of the disclosure, or pharmaceutically acceptable salt thereof, is present in a pharmaceutical composition in an amount from about 5 pg to 2,000 pg. In some embodiments, a antibody / compound of the disclosure and / or (pharmaceutically acceptable) salt thereof is present in a pharmaceutical composition in an amount from about 5 pg to 1 ,000 pg. In some embodiments, a antibody / compound of the disclosure and / or (pharmaceutically acceptable) salt thereof is present in a pharmaceutical composition in an amount from about 5 pg to 500 pg. In some embodiments, a antibody / compound of the disclosure and / or (pharmaceutically acceptable) salt thereof is present in a pharmaceutical composition in an amount from about 5 pg to 250 pg. In some embodiments, a antibody / compound of the disclosure and / or (pharmaceutically acceptable) salt thereof is present in a pharmaceutical composition in an amount from about 5 pg to 100 pg. In some embodiments, a antibody / compound of the disclosure and / or (pharmaceutically acceptable) salt thereof is present in a pharmaceutical composition in an amount from about 5 pg to 50 pg.

[0125] In some embodiments, a antibody / compound of the disclosure and / or (pharmaceutically acceptable) salt thereof is present in a pharmaceutical composition in an amount from about 1 mg to about 5,000 mg. In some embodiments, a antibody / compound of the disclosure and / or (pharmaceutically acceptable) salt thereof is present in a pharmaceutical composition in an amount from about 1 mg to about 3,000 mg. In some embodiments, a antibody / compound of the disclosure and / or (pharmaceutically acceptable) salt thereof is present in a pharmaceutical composition in an amount from about 1 mg to about 2,000 mg. In some embodiments, a antibody / compound of the disclosure and / or (pharmaceutically acceptable) salt thereof is present in a pharmaceutical composition in an amount from about 1mg to about 1 ,000 mg. In some embodiments, a antibody / compound of the disclosure and / or (pharmaceutically acceptable) salt thereof is present in a pharmaceutical composition in an amount from about 1 mg to about 500 mg. In some embodiments, a antibody / compound of the disclosure and / or (pharmaceutically acceptable) salt thereof is present in a pharmaceutical composition in an amount from about 1 mg to about 250 mg. In some embodiments, a antibody / compound of the disclosure and / or (pharmaceutically acceptable) salt thereof is present in a pharmaceutical composition in an amount from about 1 mg to about 100 mg. In some embodiments, a antibody / compound of the disclosure and / or (pharmaceutically acceptable) salt thereof is present in a pharmaceutical composition in an amount from about 1 mg to about 50 mg.

[0126] In some embodiments, a antibody / compound of the disclosure and / or (pharmaceutically acceptable) salt thereof is present in a pharmaceutical composition in an amount from about 5 mg to 2,000 mg. In some embodiments, a antibody / compound of the disclosure and / or (pharmaceutically acceptable) salt thereof is present in a pharmaceutical composition in an amount from about 5 mg to 1 ,000 mg. In some embodiments, a antibody / compound of the disclosure and / or (pharmaceutically acceptable) salt thereof is present in a pharmaceutical composition in an amount from about 5 mg to 500 mg. In some embodiments, a antibody / compound of the disclosure and / or (pharmaceutically acceptable) salt thereof is present in a pharmaceutical composition in an amount from about 5 mg to 250 mg. In some embodiments, a antibody / compound of the disclosure and / or (pharmaceutically acceptable) salt thereof is present in a pharmaceutical composition in an amount from about 5 mg to 100 mg. In some embodiments, a antibody / compound of the disclosure and / or (pharmaceutically acceptable) salt thereof is present in a pharmaceutical composition in an amount from about 5 mg to 50 mg.

[0127] In some embodiments, a antibody / compound of the disclosure and / or (pharmaceutically acceptable) salt thereof is present in a pharmaceutical composition in an amount from about 10 mg to 2,000 mg. In some embodiments, a antibody / compound of the disclosure and / or (pharmaceutically acceptable) salt thereof is present in a pharmaceutical composition in an amount from about 10 mg to 1 ,000 mg. In some embodiments, a antibody / compound of the disclosure and / or (pharmaceutically acceptable) salt thereof is present in a pharmaceutical composition in an amount from about 10 mg to 500 mg. In some embodiments, a antibody / compound of the disclosure and / or (pharmaceutically acceptable) salt thereof is present in a pharmaceutical composition in an amount from about 10 mg to 250 mg.In some embodiments, a anti body / com pound of the disclosure and / or (pharmaceutically acceptable) salt thereof is present in a pharmaceutical composition in an amount from about 10 mg to 100 mg. In some embodiments, a antibody / compound of the disclosure and / or (pharmaceutically acceptable) salt thereof is present in a pharmaceutical composition in an amount from about 10 mg to 50 mg.

[0128] In some embodiments, a antibody / compound of the disclosure and / or (pharmaceutically acceptable) salt thereof is present in a pharmaceutical composition in an amount from about 25 mg to 2,000 mg. In some embodiments, a antibody / compound of the disclosure and / or (pharmaceutically acceptable) salt thereof is present in a pharmaceutical composition in an amount from about 25 mg to 1 ,000 mg. In some embodiments, a antibody / compound of the disclosure and / or (pharmaceutically acceptable) salt thereof is present in a pharmaceutical composition in an amount from about 25 mg to 500 mg. In some embodiments, a antibody / compound of the disclosure and / or (pharmaceutically acceptable) salt thereof is present in a pharmaceutical composition in an amount from about 25 mg to 250 mg. In some embodiments, a antibody / compound of the disclosure and / or (pharmaceutically acceptable) salt thereof is present in a pharmaceutical composition in an amount from about 25 mg to 100 mg. In some embodiments, a antibody / compound of the disclosure and / or (pharmaceutically acceptable) salt thereof is present in a pharmaceutical composition in an amount from about 25 mg to 50 mg.

[0129] In some embodiments, a antibody / compound of the disclosure and / or (pharmaceutically acceptable) salt thereof is present in a pharmaceutical composition in an amount from about 50 mg to 2,000 mg. In some embodiments, a antibody / compound of the disclosure and / or (pharmaceutically acceptable) salt thereof is present in a pharmaceutical composition in an amount from about 50 mg to 1 ,000 mg. In some embodiments, a antibody / compound of the disclosure and / or (pharmaceutically acceptable) salt thereof is present in a pharmaceutical composition in an amount from about 50 mg to 500 mg. In some embodiments, a antibody / compound of the disclosure and / or (pharmaceutically acceptable) salt thereof is present in a pharmaceutical composition in an amount from about 50 mg to 250 mg. In some embodiments, a antibody / compound of the disclosure and / or (pharmaceutically acceptable) salt thereof is present in a pharmaceutical composition in an amount from about 50 mg to 100 mg.

[0130] In some embodiments, a antibody / compound of the disclosure and / or (pharmaceutically acceptable) salt thereof is present in a pharmaceutical composition in anamount from about 100 mg to 2,000 mg. In some embodiments, a antibody / compound of the disclosure and / or (pharmaceutically acceptable) salt thereof is present in a pharmaceutical composition in an amount from about 100 mg to 1 ,000 mg. In some embodiments, a antibody / compound of the disclosure and / or (pharmaceutically acceptable) salt thereof is present in a pharmaceutical composition in an amount from about 100 mg to 500 mg. In some embodiments, a antibody / compound of the disclosure and / or (pharmaceutically acceptable) salt thereof is present in a pharmaceutical composition in an amount from about 100 mg to 250 mg.

[0131] In some embodiments, a antibody / compound of the disclosure and / or (pharmaceutically acceptable) salt thereof is present in a pharmaceutical composition in an amount from about 250 mg to 2,000 mg. In some embodiments, a antibody / compound of the disclosure and / or (pharmaceutically acceptable) salt thereof is present in a pharmaceutical composition in an amount from about 250 mg to 1 ,000 mg. In some embodiments, a antibody / compound of the disclosure and / or (pharmaceutically acceptable) salt thereof is present in a pharmaceutical composition in an amount from about 250 mg to 500 mg.

[0132] In some embodiments, a antibody / compound of the disclosure and / or (pharmaceutically acceptable) salt thereof is present in a pharmaceutical composition in an amount from about 500 mg to 2,000 mg. In some embodiments, a antibody / compound of the disclosure and / or (pharmaceutically acceptable) salt thereof is present in a pharmaceutical composition in an amount from about 500 mg to 1 ,000 mg.

[0133] In some embodiments, a antibody / compound of the disclosure and / or (pharmaceutically acceptable) salt thereof is present in a pharmaceutical composition in an amount sufficient to administer about 1 pg, about 2 pg, about 3 pg, about 4 pg, about 5 pg, about 10 pg, about 15 pg, about 20 pg, about 25 pg, about 30 pg, about 35 pg, about 40 pg, about 45 pg, about 50 pg, about 60 pg, about 70 pg, about 80 pg, about 90 pg, about 100 pg, about 125 pg, about 150 pg, about 175 pg, about 200 pg, about 225 pg, about 250 pg, about 300 pg, about 350 pg, about 400 pg, about 450 pg, about 500 pg, about 550 pg, about 600 pg, about 650 pg, about 700 pg, about 750 pg, about 800 pg, about 850 pg, about 900 pg, about 1 ,000 pg, about 1 ,100 pg, about 1 ,200 pg, about 1 ,300 pg, about 1 ,400 pg, about 1,500 pg, about 1 ,600 pg, about 1 ,700 pg, about 1 ,800 pg, about 1 ,900 pg, about 2,000 pg, about 2, 100 pg, about 2,200 pg, about 2,300 pg, about 2,400 pg, about 2,500 pg, about 2,600 pg, about 2,700 pg, about 2,800 pg, about 2,900 pg, about 3,000 pg, about 3,100 pg, about 3,200 pg, about 3,300 pg, about 3,400 pg, about 3,500 pg, about 3,600 pg, about 3,700 pg, about 3,800 pg, about 3,900 pg, about 4,000 pg, about 4,100 pg, about 4,200 pg, about 4,300 pg, about4,400 pg, about 4,500 pg, about 4,600 pg, about 4,700 pg, about 4,800 pg, about 4,900 pg, 5,000 pg, about 5,100 pg, about 5,200 pg, about 5,300 pg, about 5,400 pg, about5,500 pg, about 5,600 pg, about5,700 pg, about 5,800 pg, about 5,900 pg, or about 6,000 pg per kilogram of body weight of the subject.

[0134] In some embodiments, a antibody / compound of the disclosure and / or (pharmaceutically acceptable) salt thereof is present in a pharmaceutical composition in an amount of about 1 mg, about 2 mg, about 3 mg, about 4 mg, about 5 mg, about 10 mg, about 15 mg, about 20 mg, about 25 mg, about 30 mg, about 35 mg, about 40 mg, about 45 mg, about 50 mg, about 60 mg, about 70 mg, about 80 mg, about 90 mg, about 100 mg, about 125 mg, about 150 mg, about 175 mg, about 200 mg, about 225 mg, about 250 mg, about 300 mg, about 350 mg, about 400 mg, about 450 mg, about 500 mg, about 550 mg, about 600 mg, about 650 mg, about 700 mg, about 750 mg, about 800 mg, about 850 mg, about 900 mg, about 1 ,000 mg, about 1 ,100 mg, about 1 ,200 mg, about 1 ,300 mg, about 1 ,400 mg, about 1 ,500 mg, about 1 ,600 mg, about 1 ,700 mg, about 1 ,800 mg, about 1 ,900 mg, about 2,000 mg, about 2,100 mg, about 2,200 mg, about 2,300 mg, about 2,400 mg, about 2,500 mg, about 2,600 mg, about 2,700 mg, about 2,800 mg, about 2,900 mg, about 3,000 mg, about 3,100 mg, about 3,200 mg, about 3,300 mg, about 3,400 mg, about 3,500 mg, about 3,600 mg, about 3,700 mg, about 3,800 mg, about 3,900 mg, about 4,000 mg, about 4,100 mg, about 4,200 mg, about 4,300 mg, about 4,400 mg, about 4,500 mg, about 4,600 mg, about 4,700 mg, about 4,800 mg, about 4,900 mg, or about 5,000 mg.

[0135] In some embodiments, a pharmaceutical composition comprises an antibody / compoundof the disclosure or pharmaceutically acceptable salt thereof in an amount from about 0.1 mg / mL to about 500 mg / ml_. In some embodiments, a pharmaceutical composition comprises an antibody / compoundof the disclosure or pharmaceutically acceptable salt thereof in an amount from about 0.1 mg / mL to about 100 mg / mL. In some embodiments, a pharmaceutical composition comprises an antibody / compoundof the disclosure or pharmaceutically acceptable salt thereof in an amount from about 0.1 mg / mL to about 50 mg / mL. In some embodiments, a pharmaceutical composition comprises an antibody / compoundof the disclosure or pharmaceutically acceptable salt thereof in an amount from about 0.1 mg / mL to about 5 mg / mL. In some embodiments, a pharmaceutical composition comprises an antibody / compoundof the disclosure or pharmaceutically acceptable salt thereof in an amount from about 0.1 mg / mL to about 2 mg / mL. In some embodiments, a pharmaceutical composition comprises an antibody / compoundof the disclosure or pharmaceutically acceptablesalt thereof in an amount from about 0.1 mg / mL to about 1 mg / mL. In some embodiments, a pharmaceutical composition comprises an antibody / compoundof the disclosure or pharmaceutically acceptable salt thereof in an amount from about 0.1 mg / mL to about 0.5 mg / mL.Parenteral Dosage Forms

[0136] In some or any embodiments, provided are parenteral dosage forms. Parenteral dosage forms can be administered to subjects by various routes including, but not limited to, subcutaneous, intravenous (including bolus injection), intramuscular, and intra-arterial. Because their administration typically bypasses subjects' natural defenses against contaminants, parenteral dosage forms are typically, sterile or capable of being sterilized prior to administration to a subject. In some or any embodiments, parenteral dosage forms include, but are not limited to, solutions ready for injection, dry products ready to be dissolved or suspended in a pharmaceutically acceptable vehicle for injection, suspensions ready for injection, and emulsions. In addition, with respect to subcutaneous delivery, a pen delivery device readily has applications in delivering a pharmaceutical composition of the present disclosure. Such a pen delivery device can be reusable or disposable. A reusable pen delivery device generally utilizes a replaceable cartridge that contains a pharmaceutical composition. Once all of the pharmaceutical composition within the cartridge has been administered and the cartridge is empty, the empty cartridge can readily be discarded and replaced with a new cartridge that contains the pharmaceutical composition. The pen delivery device can then be reused. In a disposable pen delivery device, there is no replaceable cartridge. Rather, the disposable pen delivery device comes prefilled with the pharmaceutical composition held in a reservoir within the device. Once the reservoir is emptied of the pharmaceutical composition, the entire device is discarded. Numerous reusable pen and autoinjector delivery devices have applications in the subcutaneous delivery of a pharmaceutical composition of the present disclosure. Examples include, but are not limited to AUTOPEN™ (Owen Mumford, Inc., Woodstock, UK), DISETRONIC™ pen (Disetronic Medical Systems, Bergdorf, Switzerland), HUMALOG MIX 75 / 25™ pen, HUMALOG™ pen, HUMALIN 70 / 30™ pen (Eli Lilly and Co., Indianapolis, IN), NOVOPEN™ I, II and III (Novo Nordisk, Copenhagen, Denmark), NOVOPEN JUNIOR™ (Novo Nordisk, Copenhagen, Denmark), BD™ pen (Becton Dickinson, Franklin Lakes, NJ), OPTIPEN™, OPTIPEN PRO™, OPTIPEN STARLET™, and OPTICLIK™ (sanofi-aventis, Frankfurt, Germany), to name only a few. Examples of disposable pen delivery devices havingapplications in subcutaneous delivery of a pharmaceutical composition of the present disclosure include, but are not limited to the SOLOSTAR™ pen (sanofi-aventis), the FLEXPEN™ (Novo Nordisk), and the KWIKPEN™ (Eli Lilly), the SURECLICK™ Autoinjector (Amgen, Thousand Oaks, CA), the PENLET™ (Haselmeier, Stuttgart, Germany), the EPIPEN (Dey, L.P.), and the HUMIRA™ Pen (Abbott Labs, Abbott Park IL), to name only a few. The injectable preparations may include dosage forms for intravenous, subcutaneous, intracutaneous and intramuscular injections, drip infusions, etc. These injectable preparations may be prepared by methods publicly known. For example, the injectable preparations may be prepared, e.g., by dissolving, suspending or emulsifying the antigen binding molecule or its salt described above in a sterile aqueous medium or an oily medium conventionally used for injections. As the aqueous medium for injections, there are, for example, physiological saline, an isotonic solution containing glucose and other auxiliary agents, etc., which may be used in combination with an appropriate solubilizing agent such as an alcohol (e.g., ethanol), a polyalcohol (e.g., propylene glycol, polyethylene glycol), a nonionic surfactant [e.g., polysorbate 80, HCO-50 (polyoxyethylene (50 mol) adduct of hydrogenated castor oil)], etc. As the oily medium, there are employed, e.g., sesame oil, soybean oil, etc., which may be used in combination with a solubilizing agent such as benzyl benzoate, benzyl alcohol, etc. The injection thus prepared is preferably filled in an appropriate ampoule.

[0137] Suitable vehicles that can be used to provide parenteral dosage forms are well known to those skilled in the art. In some or any embodiments, suitable vehicles include, but are not limited to: Water for Injection USP; aqueous vehicles such as, but not limited to, Sodium Chloride Injection, Ringer's Injection, Dextrose Injection, Dextrose and Sodium Chloride Injection, and Lactated Ringer's Injection; water miscible vehicles such as, but not limited to, ethyl alcohol, polyethylene glycol, and polypropylene glycol; and non-aqueous vehicles such as, but not limited to, com oil, cottonseed oil, peanut oil, sesame oil, ethyl oleate, isopropyl myristate, and benzyl benzoate.

[0138] Compounds that increase the solubility of one or more of the active ingredients disclosed herein can also be incorporated into the parenteral dosage forms.Transdermal, Topical & Mucosal Dosage Forms

[0139] Also provided are transdermal, topical, and mucosal dosage forms. Transdermal, topical, and mucosal dosage forms include, but are not limited to, ophthalmic solutions, sprays, aerosols, creams, lotions, ointments, gels, solutions, emulsions, suspensions, or other forms known to one of skill in the art. See, e.g., Remington: The Science and Practice of Pharmacy;Pharmaceutical Press; 22 edition (Sep. 15, 2012); and Introduction to Pharmaceutical Dosage Forms, 4th ed., Lea & Febiger, Philadelphia (1985). Dosage forms suitable for treating mucosal tissues within the oral cavity can be formulated as mouthwashes or as oral gels. Further, transdermal dosage forms include “reservoir type” or “matrix type” patches, which can be applied to the skin and worn for a specific period of time to permit the penetration of a desired amount of active ingredients.Oral Dosage Forms and Buccal Dosage Forms

[0140] The pharmaceutical compositions for oral or parenteral use described above may be prepared into dosage forms in a unit dose suited to fit a dose of the active ingredients. Such dosage forms in a unit dose include, for example, tablets, pills, capsules, injections (ampoules), suppositories, etc. The amount of the aforesaid antigen binding molecule contained is generally about 1 to about 1000 mg per dosage form in a unit dose; especially in the form of injection, it is preferred that the aforesaid antigen binding molecule is contained in about 1 to about 100 mg and in about 10 to about 250 mg for the other dosage forms. In some embodiments, the unit dosage can be as much as about 750 mg, 800 mg, 900 mg, or 1000 mg.DOSAGES

[0141] The dose of antibody administered to a patient may vary depending upon the age and the size of the patient, target disease, conditions, route of administration, and the like. The preferred dose is typically calculated according to body weight or body surface area. When a antibody of the present disclosure is used for therapeutic purposes in an adult patient, it may be advantageous to intravenously administer the antibody of the present disclosure normally at a single dose of about 0.01 to about 20 mg / kg body weight, more preferably about 0.02 to about 7, about 0.03 to about 5, or about 0.05 to about 3 mg / kg body weight. Depending on the severity of the condition, the frequency and the duration of the treatment can be adjusted. Effective dosages and schedules for administering an antibody may be determined empirically; for example, patient progress can be monitored by periodic assessment, and the dose adjusted accordingly. Moreover, interspecies scaling of dosages can be performed using well-known methods in the art (e.g., Mordenti et al., 1991 , Pharmaceut. Res. 8:1351).

[0142] In one embodiment, the antibodies and compositions of the disclosure may be administered in an amount related to the weight of the patient in need thereof. For example, the Antibody may be administered in an amount of about: 0.1 mg / kg to about 50 mg / kg, 0.1 mg / kg to about 40 mg / kg, 0.1 mg / kg to about 30 mg / kg, 0.1 mg / kg to about 25 mg / kg, 0.1 mg / kg to about 20 mg / kg, 0.1 mg / kg to about 15 mg / kg, 0.1 mg / kg to about 10 mg / kg, 0.1 mg / kg to about7.5 mg / kg, 0.1 mg / kg to about 5 mg / kg, 0.1 mg / kg to about 2.5 mg / kg, or about 0.1 mg / kg to about 1 mg / kg. The Antibody may be administered in an amount of about: 0.5 mg / kg to about 50 mg / kg, 0.5 mg / kg to about 40 mg / kg, 0.5 mg / kg to about 30 mg / kg, 0.5 mg / kg to about 25 mg / kg, 0.5 mg / kg to about 20 mg / kg, 0.5 mg / kg to about 15 mg / kg, 0.5 mg / kg to about 10 mg / kg, 0.5 mg / kg to about 7.5 mg / kg, 0.5 mg / kg to about 5 mg / kg, 0.5 mg / kg to about 2.5 mg / kg, or about 0.5 mg / kg to about 1 mg / kg. The Antibody may be administered in an amount of about 0.5 mg / kg to about 5 mg / kg or about 0.1 mg / kg to about 10 mg / kg. The Antibody may be administered in an amount of about 0.1 mg / kg to about 20 mg / kg or about 0.1 mg / kg to about 30 mg / kg. As a non limiting example, if particularly effective, the antibody of the disclosure may be administered by injection at a dose comprised between 50mg / kg and 5 pg / kg of subject, preferably at a dose comprised between 10 mg / kg and 100 pg / kg and most preferably at a dose comprised between of 2.5 mg / kg and 500 pg / kg.

[0143] The antibodies may be administered alone or in the combination at an amount of about: 1 mg, 5 mg, 10 mg, 15 mg, 20 mg, 25 mg, 30 mg, 40 mg, 50 mg, 60 mg, 70 mg, 75 mg, 80 mg, 90 mg, 100 mg, 150 mg, or 200 mg. The Antibody may be administered alone or in the combination at an amount of about: 250 mg, 300 mg, 400 mg, 500 mg, 600 mg, 700 mg, 800 mg, 900 mg, 1000 mg, 1100 mg, 1200 mg, 1300 mg, 1400 mg, 1500 mg, 1600 mg, 1700 mg, 1800 mg, 1900 mg, or 2000 mg. The Antibody may be administered alone or in the combination at an amount of about 1000 mg to about 2000 mg. The Antibody may be administered alone or in the combination at an amount of about: 1 mg to about 10 mg, 10 mg to about 20 mg, 25 mg to about 50 mg, 30 mg to about 60 mg, 40 mg to about 50 mg, 50 mg to about 100 mg, 75 mg to about 150 mg, 100 mg to about 200 mg, 200 mg to about 500 mg, 500 mg to about 1000 mg, 1000 mg to about 1200 mg, 1000 mg to about 1500 mg, 1200 mg to about 1500 mg, or 1500 to about 2000 mg.

[0144] A synergistic effect of a combination described herein may permit the use of lower dosages of one or more of the components of the combination (e.g., an antibody of the disclosure with a GLP-1 RA). A synergistic effect may permit less frequent administration of at least one of the administered therapies to a subject with a disease, disorder, or condition described herein. Such lower dosages and reduced frequency of administration may reduce the toxicity associated with the administration of at least one of the therapies to a subject without reducing the efficacy of the treatment. A synergistic effect as described herein avoid or reduce adverse or unwanted side effects associated with the use of any therapy.NUCLEIC ACIDS, VECTORS AND RECOMBINANT HOST CELLS

[0145] The present disclosure also provides nucleic acids encoding for the antibodies as described above, or parts thereof. Said nucleic acid may be RNA or DNA, preferably DNA. According to a preferred embodiment, the nucleic acid encoding the antibody of the disclosure is operatively linked to a gene expression sequence, which directs the expression of the nucleic acid within a prokarotic or an eukaryotic cell, preferably within an eukaryotic cell. The "gene expression sequence" is any regulatory nucleotide sequence, such as a promoter sequence or promoter-enhancer combination, which facilitates the efficient transcription and translation of the antibody nucleic acid to which it is operatively linked. The gene expression sequence may, for example, be a mammalian or viral promoter, such as a constitutive or inducible promoter.

[0146] Constitutive mammalian promoters include, but are not limited to, the promoters for the following genes: hypoxanthine phosphoribosyl transferase (HPTR), adenosine deaminase, pyruvate kinase, beta. -actin promoter, muscle creatine kinase promoter, human elongation factor promoter and other constitutive promoters. Exemplary viral promoters which function constitutively in eukaryotic cells include, for example, promoters from the simian virus (e.g., SV40), papilloma virus, adenovirus, human immunodeficiency virus (HIV), cytomegalovirus (CMV), Rous sarcoma virus (RSV), hepatitis B virus (HBV), the long terminal repeats (LTR) of Moloney leukemia virus and other retroviruses, and the thymidine kinase promoter of herpes simplex virus. Other constitutive promoters are known to those of ordinary skill in the art. The promoters useful as gene expression sequences of the disclosure also include inducible promoters. Inducible promoters are expressed in the presence of an inducing agent. For example, the metallothione in promoter is induced to promote transcription and translation in the presence of certain metal ions. Others inducible promoters are known to those of ordinary skill in the art.

[0147] In general, the gene expression sequence shall include, as necessary, 5' nontranscribing and 5' non-translating sequences involved with the initiation of transcription and translation, respectively, such as a TATA box, capping sequence, CAAT sequence, and the like. Especially, such 5' non-transcribing sequences will include a promoter region which includes a promoter sequence for transcriptional control of the operationally joined nucleic acid. The gene expression sequences optionally include enhancer sequences or upstream activator sequences as desired. As used herein, the nucleic acid sequence encoding the antibody of the disclosure and the gene expression sequence are said to be " operationally linked" when they are covalently linked in such a way as to place the expression or transcription and / or translation ofthe antibody of the disclosure coding sequence under the influence or control of the gene expression sequence.

[0148] Two DNA sequences are said to be operationally linked if induction of a promoter in the 5' gene expression sequence results in the transcription of the antibody of the disclosure and if the nature of the linkage between the two DNA sequences does not (1) result in the introduction of a frame-shift mutation, (2) interfere with the ability of the promoter region to direct the transcription of the antibody of the disclosure, or (3) interfere with the ability of the corresponding RNA transcript to be translated into a protein. Thus, a gene expression sequence would be operationally linked to a nucleic acid sequence coding for the antibody of the disclosure if the gene expression sequence were capable of effecting transcription of that nucleic acid sequence such that the resulting transcript is translated into the desired polypeptide.

[0149] Advantageously, said nucleic acid sequence comprises an intron, since pre- mRNA molecules has often been demonstrated to improve production yields of recombinant molecules. Any sequences of intron may be sued, and as an example, one can cite tone ones disclosed in ZAGO et al. (Biotechnol. Appl. Biochem., vol.52(Pt 3), p: 191-8, 2009) and in CAMPOS- DA- PAZ et al. (Mol. Biotechnol., vol.39(2), p:155-8, 2008).

[0150] The nucleic acid coding for the antibody of the disclosure may be delivered in vivo alone or in association with a vector. Accordingly, the present disclosure also provides vectors comprising one or more nucleic acid as described above. The term “vector,” as used herein, refers to a nucleic acid molecule capable of propagating another nucleic acid to which it is linked. The term includes the vector as a self- replicating nucleic acid structure as well as the vector incorporated into the genome of a host cell into which it has been introduced. Certain vectors are capable of directing the expression of nucleic acids to which they are operatively linked. Such vectors are referred to herein as “expression vectors.” The term “transfected” or “transformed” or “transduced” as used herein refers to a process by which exogenous nucleic acid is transferred or introduced into the host cell. A “transfected” or “transformed” or “transduced” cell is one which has been transfected, transformed or transduced with exogenous nucleic acid. The cell includes the primary subject cell and its progeny. In its broadest sense, a "vector" is any vehicle capable of facilitating the transfer of the nucleic acid coding for the antibody of the disclosure to the cells. Preferably, the vector transports the nucleic acid to cells with reduced degradation relative to the extent of degradation that would result in the absence of the vector. In general, the vectors useful in the disclosure include, but are not limited to,plasmids, cosmids, phagmids, episomes, artificial chromosomes, viruses, other vehicles derived from viral or bacterial sources that have been manipulated by the insertion or incorporation of the antibody nucleic acid sequences.

[0151] Plasmid vectors are a preferred type of vector and have been extensively described in the art and are well known to those of skill in the art. See e.g., SANBROOK et al., "Molecular Cloning: A Laboratory Manual," Fourth Edition, Cold Spring Harbor Laboratory Press, 2012. Not limiting examples of plasmids include pBR322, pUC18, pUC19, pRC / CMV, SV40, and pBlueScript, and other plasmids are well known to those of ordinary skill in the art. Additionally, plasmids may be custom designed using restriction enzymes and ligation reactions to remove and add specific fragments of DNA. Preferably, the nucleic acid vector can include selectable markers that are active both in bacteria and in mammalian cells.

[0152] In some embodiments, the present disclosure relates to a host cell genetically engineered with the nucleic acid or with the vector described previously. As used herein, the term "host cell genetically engineered" relates to host cells which have been transduced, transformed or transfected with the nucleic acid or with the vector described previously. As representative examples of appropriate host cells, one can cite bacterial cells, such as E. coli, fungal cells such as yeast, insect cells such as Sf9, animal cells such as CHO or COS, plant cells, etc. The selection of an appropriate host is deemed to be within the scope of those skilled in the art from the teachings herein. Preferably, the host cell genetically engineered is an animal cell, and most preferably CHO-S cell (INVITROGEN, cat N° 11619-012). Chinese hamster ovary (CHO) cells are frequently used in the biopharmaceutical industry for the manufacture of biologies such as recombinant proteins, antibodies, peptibodies, and receptor ligands. One of the reasons that CHO cells are often used is that these cells have an extensive safety track record for bi Nucleic acids, vectors and recombinant host cells

[0153] A further object of the disclosure relates to a pharmaceutical composition comprising the antibody as described above, a nucleic acid encoding thereof, or a vector comprising said nucleic acid, eventually associated with a pharmaceutically acceptable carrier. DEVICES

[0154] The present disclosure also provides a vessel (e.g., a vial or chromatography column) or injection device (e.g., syringe, pre-filled syringe or autoinjector) comprising an antibody (e.g., pharmaceutical formulation thereof) set forth herein. The vessel or injection device may be packaged into a kit.

[0155] An injection device is a device that introduces a substance into the body of a subject (e.g., a human) via a parenteral route, e.g., intraocular, intravitreal, intramuscular, subcutaneous or intravenous. For example, an injection device may be a syringe (e.g., prefilled with the pharmaceutical formulation, such as an auto-injector) which, for example, includes a cylinder or barrel for holding fluid to be injected (e.g., comprising an antibody or a pharmaceutical formulation thereof), a needle for piecing skin, blood vessels or other tissue for injection of the fluid; and a plunger for pushing the fluid out of the cylinder and through the needle bore and into the body of the subject.

[0156] A pharmaceutical composition provided herein may be delivered subcutaneously or intravenously with a standard needle and syringe. In addition, with respect to subcutaneous delivery, a pen delivery device readily has applications in delivering a pharmaceutical composition of the present disclosure. Such a pen delivery device can be reusable or disposable. A reusable pen delivery device generally utilizes a replaceable cartridge that contains a pharmaceutical composition. Once all of the pharmaceutical composition within the cartridge has been administered and the cartridge is empty, the empty cartridge can readily be discarded and replaced with a new cartridge that contains the pharmaceutical composition. The pen delivery device can then be reused. In a disposable pen delivery device, there is no replaceable cartridge. Rather, the disposable pen delivery device comes prefilled with the pharmaceutical composition held in a reservoir within the device. Once the reservoir is emptied of the pharmaceutical composition, the entire device is discarded. Numerous reusable pen and autoinjector delivery devices have applications in the subcutaneous delivery of a pharmaceutical composition of the present disclosure. Examples include, but are not limited to AUTOPEN™ (Owen Mumford, Inc., Woodstock, UK), DISETRONIC™ pen (Disetronic Medical Systems, Bergdorf, Switzerland), HUMALOG MIX 75 / 25™ pen, HUMALOG™ pen, HUMALIN 70 / 30™ pen (Eli Lilly and Co., Indianapolis, Ind.), NOVOPEN™ I, II and III (Novo Nordisk, Copenhagen, Denmark), NOVOPEN JUNIOR™ (Novo Nordisk, Copenhagen, Denmark), BD™ pen (Becton Dickinson, Franklin Lakes, N.J.), OPTIPEN™, OPTIPEN PRO™, OPTIPEN STARLET™, and OPTICLIK™ (sanofi-aventis, Frankfurt, Germany), to name only a few. Examples of disposable pen delivery devices having applications in subcutaneous delivery of a pharmaceutical composition of the present disclosure include, but are not limited to the SOLOSTAR™ pen (sanofi-aventis), the FLEXPEN™ (Novo Nordisk), and the KWIKPEN™ (Eli Lilly), the SURECLICK™ Autoinjector (Amgen, Thousand Oaks, Calif.), the PENLET™ (Haselmeier,Stuttgart, Germany), the EPIPEN (Dey, L.P.), and the HUMIRA™ Pen (Abbott Labs, Abbott Park III.), to name only a few.

[0157] Provided herein are also methods for administering an antibody of the present disclosure comprising introducing e.g., injecting, the molecule into the body of the subject, e.g., with an injection device.Examples

[0158] The following examples are put forth so as to provide those of ordinary skill in the art with a complete disclosure and description of how to make and use the methods and compositions of the disclosure and are not intended to limit the scope of what the inventors regard as their disclosure. Efforts have been made to ensure accuracy with respect to numbers used (e.g., amounts, temperature, etc.) but some experimental errors and deviations should be accounted for. Unless indicated otherwise, parts are parts by weight, molecular weight is average molecular weight, temperature is in degrees Centigrade, and pressure is at or near atmospheric.Example 1Heavy and light chain plasmid for each antibody were co-expressed using the Wuxian™ CHO transient expression system performed at Wuxi Biologies. Briefly, 6x106CHO cells per ml were cultured in their expression medium in 20 mL volume for 7 days after the DNA were transfected after which the supernatant were harvested for protein A purification.

[0159] lncre001 , an anti-GIP mAb (VH seq ID 7 and VL seq ID 8) was expressed as human lgG1 with wild type Fc (seq ID 9 for heavy chain and seq ID 10 for light chain). lncre002 / NM-136 lgG1 Fc null variant is a variant of IncreOl mAb / NM-136 lgG1 wild-type (VH seq ID 7, VL seq ID 8, heavy chain seq ID 11 , light chain seq ID 10) containing the following mutations: L234A, L235A, G237A, K322A and P329G in the Fc region to abolish FcRy receptor binding, was also expressed as described above. The protein from the CHO supernatant from the two consructs was harvested and purified to homogeneity using protein A column on GE AKTA andconfirmed for purity on SEC column using Agilent HPLC. The final protein is formulated in 20 mM Histidine Acetate, 0.15 M NaCI, pH5.5. Both IncreOOl and lncre002 proteins expressed very well and is 100% pure following protein A purification with no measureable aggregation (Table 1 and Figure 1).Table 1 : IncreOOl and lncre002 expression yield and SEC-HPLC purityExample 2.Target Binding for IncreOOl and lncre002.

[0160] Analysis of binding to human GIP Fc tag (Sino Biological / 15340-H02H) was performed using Octet Red 96 instrument (ForteBio). The Octet systems use propriety BLI to analyze biomolecular interaction. IncreOOl and lncre002 were immobilized on select capture CH1 biosensors at 2 ug / ml. These were incubated with varying concentration of human GIP Fc tag ranging from 4.11 to 333 nM in PBS. The experiments comprised of 5 steps: (1) Baseline acquisition (60 s); (2) Antibodies loading onto anti-human Fc biosensor (60 s); (3) Second baseline acquisition (60 s); (4) Association of interacting human GIP Fc tag for kon measurement (90 s); (5) Dissociation of interacting human GIP Fc tag for koff measurement (300 s). Data were analyzed using Octet Data Analysis Sofware 9.0 (ForteBio) and fitted to the 1 :1 binding model. Equilibrium dissociation constants (KD) were calculated by the ratio of kon to koff. Data is shown in figure 2. Both IncreOOl and lncre002 had similar binding affinity (KD) to human GIP, indicating that the mutation in the Fc region has no impact on the binding to its target, GIP. Figure 2 is the sensogram of the interaction.Example 3 Fcgamma receptor binding

[0161] Octet binding is used to test Fc-receptor binding on the wild-type and Fc null variants. Binding to the following receptors is tested:

[0162] FcgR1 (CD64)

[0163] FcgR2a (CD32a H167 and / or R167 variant)

[0164] FcR2b (CD32b)

[0165] FcgR3 (CD16a V176 and / or F176 variant)

[0166] Human Fey R interaction analysis was performed using Octet Red 96 instrument. Human FcyRI (CD64a - Sino Biological / 10256-H27H-B), FcyRIIA (CD32a - Sino Biological / 10374-H27H1-B), FcyRIIB (CD32b - Sino Biological / 10259-H27H-B) and FcyRIIIA (CD16a - Sino Biological / 10389-H27H1-B) proteins were immobilized on anti-his tag biosensors and incubated with varying concentration of the mAbs (lncre001 and lncre002) ranging from 3000 nM to 4.11 nM or 1000 nM to 1.37 nM. The experiment comprised of 5 steps: (1) Baseline acquisition (30 s); (2) human FcyRs loading to anti-his tag biosensor (60 s); (3) Second baseline acquisition (30 s); (4) Association of interacting mAbs for kon measurement (200 s); (5) Dissociation of interaction mAbs for koff measurement (300 s). Data were analyzed using Octet Data Analysis Software 9.0 (ForteBio) and fitted to the 1 :1 binding model. Equilibrium dissociation constants (KD) were calculated by the ratio of kon to koff. The binding affinity of lncre001 and lncre002 is reported in Table 2 and the sensograms for the interaction is shown in figure 3.Table 2: Kd (in M) of lncre001 and lncre002 against different FcyRs.

[0167] The results show that the lncre002 carrying all null mutations does not bind any of these Fc receptors.Example 4Target binding using SPR

[0168] Analysis of binding to GIP peptide (Phoenixpeptide / 027-020) was performed using Biacore T200 (Cytiva). The Biacore T200 system uses SPR to analyze biomolecularinteraction. IncreOOl and lncre002 were immobilized on anti-human on the CM5 sensor chip at 10 ug / ml. These were incubated with varying concentration of GIP peptide ranging from 1 .23 to 300 nM in HBS-EP+ buffer (10mM HEPES pH7.4, 150mM NaCI, 3mM EDTA, 0.05% Tween 20) at 25°C. IncreOOl and lncre002 was captured for 20s at 10ul_ / min. The association time was 90s at 30uL / min followed dissociation for 450s. Both IncreOOl and lncre002 had similar binding affinity (KD) to human GIP peptide further confirming that the mutation in the Fc region had no impact on binding. FIG. 4 is the sensogram of the interaction.Example 5 Epitope mapping of lncre001 / 002

[0169] To map out the epitopes where lncre001 / 002 binds to human GIP, a series of 13 different peptides were synthesized as shown in FIG. 5. Each peptide was coated on the ELISA plate at a concentration of 50 ug / mL in duplicate wells overnight at 4 °C, washed with PBST (137 mM NaCI, 2.7 mM KCI, 4.3 mM Na2HPO4, 0.05% Tween-20, pH7.4) and blocked with PBST pH7.4 with 3% BSA. lncre002 was added at 100 ug / mL and 150 ug / mL and incubated for 1 hour at 25 °C, washed 4x and blocked with PBST pH7.4 plus 3% BSA. Anti-human Fc HRP at 0.5ug / mL was then added and incubated for 1 hour. The plate was washed 4x and OD of 450nm - 620nm was read to determine the binding activity.

[0170] The data on FIG. 6 and FIG. 7 showed that the epitope for lncre001 / 002 mapped to the C-terminus of the human GIP peptide (KNDWKHNITQ (SEQ ID NO: 16)). Alanine scan of this C-terminus peptide shows that the peptide K_DWKH (SEQ ID NO: 31) closer to the C- terminus of human GIP is critical for binding.Example 6 lncre002 was engineered with half-life extension mutations in the Fc region

[0171] Two additional lncre002 variants were made by introducing Fc mutations on top of the L234A, L235A, G237A, K322A, and P329G mutations: lncre004 containing the YTE (M252Y, S254T and T256E) mutations (VH seq ID 7, VL seq ID 8, heavy chain seq ID 12, light chain seq ID 10) and lncre005 containing the LS (M428L and N434S) mutations (VH seq ID 7, VL seq ID 8, heavy chain seq ID 13, light chain seq ID 10). These two variants were produced in CHO cells and purified as described in example 1. FIG. 8 shows the purity of lncre004 and lncre005.

[0172] The purified proteins were tested for FcRn binding at pH 6.0 and pH7.4 using Biacore 8K (Cytiva). The antibodies (lncre002, lncre004 and lncre005) immobilized on the CM5 chip via anti-human. The running buffer used was either 1 *PBST (pH 6.0) or 1*PBST (pH 7.4). Human human FcRn (Wuxi Biologies, MH025-D09H) at pH 6.0 or pH 7.4 were used as analyte at concentration ranging from 12 to 6000 nM. 60 sec association and 90 sec dissociation time was used with flow rate of 30 pL / min.

[0173] FIG. 9 and FIG. 10 show that lncre004 and ncre005 containing the YTE and LS mutations respectively binds to human FcRn tenfold better than lncre002, the parental molecule which does not contain the YTE or the LS mutations at pH 6.0. At pH 7.4, none of these mutations exhibit measurable affinity. lncre004 and lncre005 exhibited characteristics of antibody with extended half-life due to their enhanced binding at pH 6.0 and no binding at pH 7.4. lncre004 and lncre005 are expected to bind tighter to FcRn at low pH in the cells so they don’t get degraded but gets recycle readily when the receptors were transported to cell surface where the pH is around 7. It appears that the half life extension mutations retain the desired property when engineered on an Fc that already have other mutations to disabled Fey receptors binding.Example 7 Murinization of lncre002 with engineered mu Fc

[0174] Murinization was performed on lncre001 / lncre002 variable domains by grafting the heavy chain CDRs of lncre001 / lncre002 into mouse heavy chain germline, IGHV1-66*01 with several framework backmutation to improve binding. Similarly, the light chain CDRs of lncre001 / lncre002 was grafted into mouse kappa light chain germline, IGKV10-94*01 with no backmutation. The murinized version lncre002, also known as lncre003 (heavy chain seq ID 14, light chain seq ID 15), was fused to the mouse lgG2a constant domain containing the following mutation to disable Fc function: L234A, L235A, G237A, K322A and P329G. lncre003 was produced in CHO cells and purified as described in example 1 with yield of 1.5 gram per L and purity of >90%.

[0175] Analysis of binding of lncre003 to mouse GIF peptide (Phoenix peptide / 027-27) was performed using Biacore 8K (Cytiva). The Biacore 8K system uses SPR to analyze biomolecular interaction. lncre003 was immobilized on anti-mouse on the CM5 sensor chip at 5 ug / ml. It was incubated with varying concentration of mouse GIP peptide ranging from 0.41 to 300 nM in HBS-EP+ buffer (10mM HEPES pH7.4, 150mM NaCI, 3mM EDTA, 0.05% Tween 20)at 25°C. lncre003 was captured for 120s at 10uL / min. The association time was 90s at 30uL / min followed dissociation for 450s. lncre003 had a binding affinity (KD) of 1.24 nM to mouse GIP peptide similar to lncre02 KD affinity of 1.2 nM for human GIP peptide. FIG. 11 showed the sensogram of the interaction.Example 12Murine Fcgamma receptor binding using Octet

[0176] Mouse Fey R interaction analysis was performed using Octet Red 96 instrument. Mouse FcyRI (CD64a - AcroBiosystems / CD4-M5227), FcyRIIB (AcroBiosystems / CDB-M52H7), FcyRIII (AcroBiosystems / CDA-M52H8) and FcyRIV (CD16-2 - AcroBiosystems / FC4-M52H3) proteins were immobilized on anti-his tag biosensors and incubated with varying concentration of lncre003 ranging from 1000 nM to 4.11 nM. The experiment comprised of 5 steps: (1) Baseline acquisition (30 s); (2) human FcyRs loading to anti-his tag biosensor (60 s); (3) Second baseline acquisition (30 s); (4) Association of interacting mAbs for kon measurement (200 s); (5) Dissociation of interaction mAbs for koff measurement (300 s). Data were analyzed using Octet Data Analysis Software 9.0 (ForteBio) and fitted to the 1 :1 binding model. Equilibrium dissociation constants (KD) were calculated by the ratio of kon to koff. Unlike the lgG2a control, lncre003 does not show any binding to the 4 different receptors. FIG. 12 showed the sensograms.

[0177] Embodiments or descriptions that include “or” between one or more members of a group are considered satisfied if one, more than one, or all of the group members are present in, employed in, or otherwise relevant to a given product or process unless indicated to the contrary or otherwise evident from the context. The disclosure includes embodiments in which exactly one member of the group is present in, employed in, or otherwise relevant to a given product or process. The disclosure includes embodiments in which more than one, or all of the group members are present in, employed in, or otherwise relevant to a given product or process. Furthermore, the disclosure encompasses all variations, combinations, and permutations in which one or more limitations, elements, clauses, and descriptive terms from one or more of the listed claims is introduced into another claim. For example, any claim that is dependent on another claim can be modified to include one or more limitations found in any other claims that is dependent on the same base claim. Where elements are presented as lists, e.g., in Markush group format, each subgroup of the elements is also disclosed, and anyelement(s) can be removed from the group. It should it be understood that, in general, where the disclosure, or aspects of the disclosure, is / are referred to as comprising particular elements and / or features, certain embodiments of the disclosure or aspects of the disclosure consist, or consist essentially of, such elements and / or features. For purposes of simplicity, those embodiments have not been specifically set forth in haec verba herein. It is also noted that the terms “comprising” and “containing” are intended to be open and permits the inclusion of additional elements or steps. Where ranges are given, endpoints are included. Furthermore, unless otherwise indicated or otherwise evident from the context and understanding of one of ordinary skill in the art, values that are expressed as ranges can assume any specific value or sub-range within the stated ranges in different embodiments of the disclosure, to the tenth of the unit of the lower limit of the range, unless the context clearly dictates otherwise.Example 13 Pharmacodynamic Efficacy of INC-118 Demonstrated in Murine Oral Glucose Tolerance Test Model

[0178] Study Purpose:

[0179] The purpose of this study was to evaluate the effect of GIP mAb-INC 118 on the insulin response to glucose via oral glucose tolerance test (OGTT) in C57BL / 6J mice. This study was conducted by WuXi AppTec at its facility, an AAALAC accredited facility and all animal study procedures were approved by the Institutional Animal Care and Use Committee (IACUC) of WuXi AppTec.

[0180] Methods:

[0181] Male C57BL / 6J mice (8 weeks) were purchased from GemPharmatech Co., Ltd.

[0182] Animals were acclimatized for 1 week prior to the experiment.

[0183] A total of 12 animals were used for this study.

[0184] The dosing volume was 10 mL / kg.

[0185] PBS (study drug dilution vehicle) acted as the control

[0186] The design, dosage and regimen of the tested compounds are outlined in the following table:

[0187] Table 3: Group and Design InformationOralSaline Pre Termination Food Treatment Treatment GlucoseGroup N Animals Treatment Pre Bleed Schedule & Withdrawal 1 (IP) 1 Schedule Tolerance(IP) CollectionsTestDay 0 1 = 0 mininsu in 118 (50 mg / kg) oo samp ing30min

[0188] Summary of Study Design:

[0189] Blood samples were collected for insulin detection on Day -1 , and at T=0 min, 15 min, 30 min after oral glucose administration on Day 0. Blood samples were collected via tail vein into tubes pre-coated with EDTA-K2 for plasma preparation. Plasma samples were obtained by centrifugation at 4000 rpm for 10 minutes at 4°C, and stored at -80°C for insulin measurement. All blood glucose was measured using ACCU-CHEK Active Blood Glucose Meter, with ACCU-CHEK Active Test Strips.

[0190] Study Results:

[0191] The effect of GIP antagonist mAb-INC-118 on insulin response to glucose via oral glucose tolerance test (OGTT) was evaluated in C57BL / 6J mice. The results showed that, compared to the PBS treatment group, treatment with GIP mAb-INC 118 (50 mg / kg) increased blood glucose levels post oral glucose administration (FIGs. 13-16), and decreased plasma insulin post oral glucose administration (FIG. 17 and FIG. 18).

[0192] Conclusion:

[0193] In this study, the expected pharmacodynamic reduction of insulin response to an oral glucose tolerance test in C57BL / 6j mice following treatment with anti-GIP mAb-INC 118 is confirmed.

[0194] This application refers to various issued patents, published patent applications, journal articles, and other publications, all of which are incorporated herein by reference. If there is a conflict between any of the incorporated references and the instant specification, the specification shall control. In addition, any particular embodiment of the present disclosure that falls within the prior art may be explicitly excluded from any one or more of the embodiments. Because such embodiments are deemed to be known to one of ordinary skill in the art, they may be excluded even if the exclusion is not set forth explicitly herein. Any particular embodiment of the disclosure can be excluded from any embodiment, for any reason, whether or not related to the existence of prior art. Those skilled in the art will recognize or be able to ascertain using no more than routine experimentation many equivalents to the specificembodiments described herein. The scope of the present embodiments described herein is not intended to be limited to the above Description, but rather is as set forth in the appended embodiments. Those of ordinary skill in the art will appreciate that various changes and modifications to this description may be made without departing from the spirit or scope of the present disclosure, as defined in the embodiments explicitly disclosed above and in the claims.

Claims

What is claimed is:

1. An antibody comprising HC-CDR1 of SEQ ID NO:1 , HC-CDR2 of SEQ ID NO:2, HC- CDR3 of SEQ ID NO:3, LC-CDR1 of SEQ ID NO:4, LC-CDR1 of SEQ ID NO:5, and LC-CDR3 of SEQ ID NO:6, and an Fc variant of a wild-type human IgG Fc region, said Fc variant comprising (i) at least one amino acid substitutions at a position selected from L234A, L235A, G237A, K322A, and P329G and / or (ii) at least one amino substitution selected from M252Y, S254T, T256E, M428L, and N434S, wherein the residues are numbered according to the EU index.

2. The antibody of claim 1 , wherein the Fc variant comprises (i) at least two, at least three, at least four, or all five amino acid substitutions at a position selected from L234A, L235A, G237A, K322A, P329G and / or (ii) the three M252Y, S254T and T256E substitutions or the two M428L and N434S substitutions.

3. The antibody of any one of claims 1 and 2, wherein the IgG is a human IgG 1 .

4. The antibody of any one of claims 1 to 3, wherein the antibody comprises a heavy chain variable region of SEQ ID 7: and a light chain variable region of SEQ ID NO:8.

5. The antibody of any one of claims 1 to 4, wherein the antibody comprises a heavy chain constant region of SEQ ID NO selected from SEQ ID NO: 9, 11 , 12, and 13, preferably in combination with a light chain sequence of SEQ ID NO: 10.

6. A pharmaceutical composition comprising an antibody of any one of claims 1 to 5 and a pharmaceutically acceptable carrier or dilutent.

7. The pharmaceutical composition of claim 6, further comprising another therapeutic agent.

8. The pharmaceutical composition of claim 7, wherein the other therapeutic agent is an agent that treats obesity and / or an obesity-associated disease or disorder.

9. The pharmaceutical composition of any one of claims 7 and 8, wherein the other therapeutic agent is selected from GIP / GLP1 dual agonists; GLP1 / amylin dual agonists; GLP1+Glucagon dual agonists; Gl PR antagonist / GLP1 agonist; GIP antagonists; appetite suppressants, lipid lowering agents, anti-diabetec agents, hypoglycemic agents, triglyceride lowering agents.

10. A nucleic acid encoding an antibody of any one of claims 1 to 5.

11. A cell comprising a nucleic acid of claim 10.

12. A method of making an antibody of any one of claims 1 to 5, said method comprising; culturing a host cell comprising a nucleic acid encoding the antibody, and culturing the cell under conditions that allow production of the antibody.

13. A method of treating a disease or disorder by administering a therapeutically effective amount of an antibody to any one of claims 1 to 5 or a composition of any one of claims 6 to 9 to a subject in need thereof.

14. The method of claim 13, wherein the disease or disorder is obesity and / or an obesity- associated disease or disorder.

15. The method of any one of claims 13 and 14, further comprising combination treatment with behavioral therapy (diet and lifestyle modifications), other therapeutic agent(s), and / or bariatric surgery.

16. The method of claim 15, wherein the other therapeutic agent(s) is administered prior to, concurrently with, and / or subsequent to the treatment with an antibody of any one of claims 1 to 5 or pharmaceutical composition of any one of claims 6 to 9.

17. The method of claim 16, wherein the other therapeutic agent(s) is an agent that treats obesity and / or an obesity-associated disease or disorder.

18. The method of any one of claims 13 to 18, wherein the subject has a BMI of >30 kg / m2 or a BMI of >27 kg / m2 with weight-related comorbidities.

19. A method of maintaining weight loss in a subject that has lost weight, comprising administering a therapeutically effective amount of an antibody according to any one of claims 1 to 5 or a composition according to any one of claims 6 to 9 to the subject.

20. A method of preventing weight regain in a subject that has lost weight, comprising administering a therapeutically effective amount of an antibody according to any one of claims 1 to 5 or a composition according to any one of claims 6 to 9 to the subject.

21. A method of preventing the “yo-yo effect” in a subject in need thereof related to constant fluctuation in weight related to losing weight then regaining the weight (or more weight) after stopping a diet, comprising administering a therapeutically effective amount of an antibody according to any one of claims 1 to 5 or a composition according to any one of claims 6 to 9 to the subject.

22. An Fc polypeptide wherein the Fc domain comprises at least one amino acid substitution that reduces binding affinity to an Fc receptor and / or effector function comprising one or more, two or more, three or more, four or more, or all five amino acid substitutions selected from L234A, L235A, G237A, K322A, and P329G, wherein the residues are numbered according to the EU index, preferably wherein the Fc polypeptide is an antibody.

23. The Fc polypeptide of claim 22, wherein the Fc domain further comprises the three M252Y, S254T and T256E substitutions or the two M428L and N434S substitutions, whereby said substitutions extend the half-life of an antibody comprising said Fc polypeptide.

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