ALK7 combination therapy

Abstract

The problem that this invention aims to solve is to provide additional treatment options for the increasing number of patients suffering from metabolic disorders and diseases, particularly those suffering from type 2 diabetes, obesity, overweight, or a combination thereof. [Solution] The present invention relates to an anti-ALK7 receptor antibody or antigen-binding fragment used in combination with a second bioactive agent, such as a GLP-1 receptor-amyrin receptor agonist, an anti-GIP receptor antibody or its antigen-binding fragment, and / or an anti-ActRII antibody or its antigen-binding fragment, for the prevention and / or treatment of overweight, obesity, and / or diabetes.

Description

TECHNICAL FIELD

[0001] The present disclosure provides combination therapies for the treatment of diseases or disorders, such as metabolic diseases or disorders, including but not limited to overweight, obesity, and / or diabetes.

[0002] Sequence Listing This application contains a Sequence Listing submitted electronically in XML file format, which is incorporated herein by reference in its entirety. The aforementioned .XML copy created on March 24, 2026 has the file name "250013US01_SL.xml" and a size of 119,033 bytes. BACKGROUND ART

[0003] Overweight and obesity are abnormal or excessive accumulations of body fat that pose risks to an individual's overall health. A body mass index (BMI) greater than 25 is considered overweight, and a BMI greater than 30 is considered obese. Obesity is a major risk factor in numerous serious medical conditions, including type 2 diabetes and its associated comorbidities, as well as cardiovascular diseases such as heart disease and stroke, which are leading causes of death worldwide. Obesity is now recognized by the World Health Organization (WHO) as a problem that has spread to epidemic proportions even among children; in 2016, 1.9 billion adults worldwide were reported to be obese, and in 2019, 38.3 million children under the age of 5 worldwide were reported to be obese. According to the WHO, 422 million people worldwide suffer from diabetes, and 1.6 million deaths each year are directly attributable to diabetes.

[0004] Activin receptor-like kinase 7 (ALK7, also known as ACVR1C) is a single transmembrane type 1 serine / threonine kinase within the TGF-β receptor family, primarily expressed on mature adipocytes in human and relevant animal models, e.g., rodents. The signaling complex is a heterooligomer consisting of a heterotetrameric receptor complex with two type I and two type II transmembrane serine / threonine kinases, which are activated by homodimeric ligands of the TGF-β superfamily, including activin B, activin C, and activin E. In the signaling complex, the type II receptor phosphorylates and activates the type I receptor, which in turn phosphorylates, and subsequently binds to and activates, the SMAD transcription regulators SMAD2 and SMAD3. Activation of ALK7 promotes lipid storage in adipose tissue. Inhibition of ALK7, including inhibition by neutralizing antibodies, has been shown to reduce total body weight in preclinical animal models. Antibodies of ALK7 have been described and their properties explained in previous patent applications such as WO17 / 1805037, WO19 / 084249, and WO20 / 244343. [Overview of the project]

[0005] This disclosure provides additional treatment options for the increasing number of patients suffering from metabolic disorders and diseases, particularly those with type 2 diabetes, obesity, overweight, or a combination thereof.

[0006] This disclosure provides combinations such as combination therapies comprising an anti-ALK7 antibody or its antigen-binding fragment and a second bioactive agent, and the use of such combination therapies for the prevention and / or treatment of overweight, obesity, and / or diabetes, such as type 2 diabetes. In particular, this disclosure provides combination therapies comprising an anti-ALK7 antibody or its antibody-binding fragment and a second bioactive agent selected from a GLP-1 receptor agonist, a GLP-1 receptor-amyrin receptor agonist, an anti-GIP receptor antibody or its antigen-binding fragment, an anti-ActRII antibody or its antigen-binding fragment, or a combination thereof. As described herein, the inventors observed synergistic effects in mice relating to combinations of an anti-ALK7 antibody or its antigen-binding fragment and a second bioactive agent such as a GLP-1 receptor agonist, a GLP-1 receptor-amyrin receptor agonist, an anti-GIP receptor antibody or its antigen-binding fragment, or an anti-ActRII antibody or its antigen-binding fragment.

[0007] Administration of a combination of anti-ALK7 antibody or its antigen-binding fragment with a second bioactive agent such as a GLP-1 receptor agonist, GLP-1 receptor-amyrin receptor agonist, anti-GIP receptor antibody or its antigen-binding fragment, or anti-ActRII antibody or its antigen-binding fragment resulted in improved total weight loss due to fat loss, comparable to the administration of anti-ALK7 antibody or its antigen-binding fragment or the second bioactive agent alone, while maintaining lean body mass.

[0008] Accordingly, this disclosure provides combination therapies and / or compositions, etc., comprising an anti-ALK7 antibody or its antigen-binding fragment and a second bioactive agent, which are useful for methods of reducing or inducing weight loss in subjects such as human subjects, and for methods of preventing and / or treating overweight, obesity, diabetes, metabolic diseases or disorders, and cardiovascular diseases (CVD). This disclosure provides an anti-ALK7 antibody or its antigen-binding fragment for use as a pharmaceutical in combination with a second bioactive agent.

[0009] In some embodiments, the present disclosure provides an anti-ALK7 antibody or an antigen-conjugated fragment thereof for use in methods for the prevention and / or treatment of overweight, obesity, and / or diabetes, wherein the anti-ALK7 antibody or the antigen-conjugated fragment thereof is administered to a subject in combination with a second bioactive agent.

[0010] In some embodiments, the present disclosure provides a second bioactive agent for use in methods for the prevention and / or treatment of overweight, obesity, and / or diabetes, the second bioactive agent being administered to a subject in combination with an anti-ALK7 receptor antibody or its antigen-binding fragment.

[0011] Administration of a GLP-1 receptor agonist (e.g., semaglutide) alone resulted in a reduction in total body mass due to a decrease in lean body mass and body fat mass. Surprisingly, administration of an anti-ALK7 receptor antibody or its antigen-binding fragment in combination with a GLP-1 receptor agonist (e.g., semaglutide) resulted in improved total body weight loss compared to administration of the anti-ALK7 receptor antibody or its antigen-binding fragment alone or the GLP-1 receptor agonist alone. This improvement in total body weight loss was mainly due to a decrease in body fat mass, while lean body mass was maintained (see Example 5).

[0012] Administration of GLP-1 receptor-amyrin receptor agonists alone resulted in a reduction in lean body mass and a decrease in total body weight resulting from a reduction in fat mass. However, administration of an anti-ALK7 receptor antibody or its antigen-binding fragment in combination with a GLP-1 receptor-amyrin receptor agonist resulted in an improvement in total body weight loss, primarily due to fat mass loss. The reduction in lean body mass observed when a GLP-1 receptor-amyrin receptor agonist was administered alone was blunted when an anti-ALK7 receptor antibody or its antigen-binding fragment was administered concurrently (see Example 6).

[0013] Administration of anti-GIP receptor antibodies or their antigen-binding fragments alone resulted in a decrease in total body weight, due to reductions in both fat mass and lean body mass. On the other hand, administration of anti-ALK7 receptor antibodies or their antigen-binding fragments in combination with anti-GIP receptor antibodies or their antigen-binding fragments resulted in a greater decrease in total body weight and greater loss of total fat mass while maintaining lean body mass (see Example 7).

[0014] Administration of anti-ActRII antibodies or their antigen-binding fragments alone resulted in changes in body composition, leading to a decrease in fat mass and an increase in lean body mass. Administration of anti-ALK7 receptor antibodies or their antigen-binding fragments resulted in a decrease in total body weight due to total fat loss while preserving lean body mass (see Example 8).

[0015] The combination therapies described herein provide means for modulating body composition in specific ways, such as targeting fat loss while preventing lean body mass loss, or even preventing an increase in lean body mass. Such combination therapies may be useful in preventing and / or treating certain diseases and / or disorders in which a decrease in lean body mass may be undesirable when it comes to total body weight loss.

[0016] This disclosure provides an anti-ALK7 receptor antibody or its antigen-binding fragment for use in combination with a second bioactive agent for the prevention and / or treatment of overweight, obesity, and / or diabetes, wherein the second bioactive agent comprises a GLP-1 receptor agonist. In some embodiments, the anti-ALK7 receptor antibody or its antigen-binding fragment is administered (i) together with the second bioactive agent, or (ii) simultaneously with or sequentially with the second bioactive agent. In some embodiments, the GLP-1 receptor agonist has an amino acid sequence having at least 90% sequence identity with the amino acids of SEQ ID NO: 2. In some embodiments, the GLP-1 receptor agonist comprises the following structure (Formula I (SEQ ID NO: 106)): [ka] The aforementioned amino acid sequence is Sequence ID No. 2, or a pharmaceutically acceptable salt thereof. In some embodiments, the GLP-1 receptor agonist comprises the following structure (Formula II (Sequence ID No. 107)): [ka] The aforementioned amino acid sequence is Sequence ID No. 2, or a pharmaceutically acceptable salt thereof.

[0017] In some embodiments, the second bioactive agent comprises a GLP-1 receptor agonist and an amyrin receptor agonist. In some embodiments, the second bioactive agent comprises a GLP-1 receptor-amyrin receptor agonist. In some embodiments, the GLP-1 receptor-amyrin receptor agonist has an amino acid sequence having at least 90% sequence identity with the amino acids of SEQ ID NO: 3. In some embodiments, the GLP-1 receptor-amyrin receptor agonist comprises the following structure (Formula IV (SEQ ID NO: 108)): [ka] The aforementioned amino acid sequence is SEQ ID NO: 3, or a pharmaceutically acceptable salt thereof. In some embodiments, the GLP-1 receptor-amyrin receptor agonist comprises the following structure (Formula V (SEQ ID NO: 109)): [ka] The aforementioned amino acid sequence is Sequence ID No. 3, or a pharmaceutically acceptable salt thereof.

[0018] In some embodiments, the anti-ALK7 receptor antibody or its antigen-binding fragment is characterized by (i) being an antagonist, (ii) blocking the binding of activin B to the ALK7 receptor, and / or (iii) blocking the formation of the ALK7 receptor hexamer signaling complex with the ActRIIA / B type II receptor. In some embodiments, the anti-ALK7 receptor antibody or its antigen-binding fragment comprises three heavy chain CDR sequences hCDR1-3 and three light chain CDR sequences lCDR1-3. a) hCDR1 is SGNYWN (SEQ ID NO: 8) or SGSYWN (SEQ ID NO: 9) with at most 1 amino acid substitution, b) hCDR2 is YISFDGRNNYNPSLKN (SEQ ID NO: 10), YISFDGRTNYNPSLKN (SEQ ID NO: 11), YISFDGRTNYNPSLKS (SEQ ID NO: 12), YISFDGRRNYNPSLKN (SEQ ID NO: 13), YISFTGRTNYNPSLKN (SEQ ID NO: 14), YISFSGRNNYNPSLKN (SEQ ID NO: 15), or YISFSGRTNYNPSLKN (SEQ ID NO: 16) with at most 1 or 2 amino acid substitutions, c) hCDR3 is DYYGSEGFGY (SEQ ID NO: 17) or DYYGSEGFAY (SEQ ID NO: 18) with at most 1 or 2 amino acid substitutions, d) lCDR1 is KSSQSLLNSRTRKNYLA (SEQ ID NO: 19), KSSQSLLVSRTRKNYLA (SEQ ID NO: 20), or KSSQSLLYSRTRKNYLA (SEQ ID NO: 21) with at most 1 or 2 amino acid substitutions, e) lCDR2 is WTSTRES (SEQ ID NO: 22) or WASTRES (SEQ ID NO: 23) with at most 1 amino acid substitution, f) lCDR3 is KQSYNLPYT (SEQ ID NO: 24) or KRSYNLPYT (SEQ ID NO: 25) with at most 1 amino acid substitution.

[0019] In some embodiments, the anti-ALK7 receptor antibody or antigen-binding fragment thereof comprises three heavy chain CDR sequences hCDR1 to 3, and three light chain CDR sequences lCDR1 to 3, i. The three heavy chain CDR sequences are selected from the CDR sequences of any one of the heavy chain variable regions (HV) identified by SEQ ID NOs: 26 to 59, and / or, ii. The three light chain CDR sequences are selected from the CDR sequences of any one of the light chain variable regions (LV) identified by SEQ ID NOs: 60 to 84.

[0020] In some embodiments, the uses described herein comprise reducing body fat mass. In some embodiments, the uses provided herein comprise preserving lean body mass.

[0021] The drawings contained herein, consisting of the following figures, are for illustrative purposes only and are not intended to be limiting. [BRIEF DESCRIPTION OF THE DRAWINGS]

[0022] [Figure 1] Figures 1A to 1D: A) Relative change in body weight from baseline (% delta), B) Daily food intake (g), C) Relative change in fat mass from baseline (% delta), D) Relative change in lean mass from baseline (% delta). Open circles represent the control, open upward-pointing triangles represent administration of an exemplary anti-ALK7 antibody, open downward-pointing triangles represent administration of an exemplary GLP-1 receptor agonist, and open squares represent administration of the combination of the exemplary anti-ALK7 antibody and the exemplary GLP-1 receptor agonist. [Figure 2] Figures 2A to 2C: A) Fasting blood glucose (mg / dL), B) Fasting plasma insulin (ng / mL), C) Liver weight, and fat and lean mass composition (g). Open circles represent the control, open upward-pointing triangles represent administration of an exemplary anti-ALK7 antibody, open downward-pointing triangles represent administration of an exemplary GLP-1 receptor agonist, and open squares represent administration of the combination of the exemplary anti-ALK7 antibody and the exemplary GLP-1 receptor agonist. [Figure 3]Figures 3A-3D: A) Relative change in body weight from baseline (% delta), B) Daily food intake (g), C) Relative change in fat mass from baseline (% delta), D) Relative change in fat mass from baseline (% delta). White circles represent controls, upward-pointing white triangles represent administration of exemplary anti-ALK7 antibody, downward-pointing white triangles represent administration of exemplary GLP-1 receptor-amyrin receptor agonist (1 nmol / kg), downward-pointing black triangles represent administration of exemplary GLP-1 receptor-amyrin receptor agonist (3 nmol / kg), white squares represent administration of a combination of exemplary anti-ALK7 antibody and exemplary GLP-1 receptor-amyrin receptor agonist (1 nmol / kg), and black squares represent administration of a combination of exemplary anti-ALK7 antibody and exemplary GLP-1 receptor-amyrin receptor agonist (3 nmol / kg). [Figure 4] Figures 4A-4C: A) Fasting blood glucose (mg / dL), B) Fasting plasma insulin (ng / mL), C) Liver mass, and fat and lean body mass composition (g). White circles represent controls, upward-pointing white triangles represent administration of exemplary anti-ALK7 antibody, downward-pointing white triangles represent administration of exemplary GLP-1 receptor-amyrin receptor agonist (1 nmol / kg), downward-pointing black triangles represent administration of exemplary GLP-1 receptor-amyrin receptor agonist (3 nmol / kg), white squares represent administration of a combination of exemplary anti-ALK7 antibody and exemplary GLP-1 receptor-amyrin receptor agonist (1 nmol / kg), and black squares represent administration of a combination of exemplary anti-ALK7 antibody and exemplary GLP-1 receptor-amyrin receptor agonist (3 nmol / kg). [Figure 5] Figures 5A-5D: A) Relative change in body weight from baseline (% delta), B) Daily dietary intake (g), C) Relative change in fat mass from baseline (% delta), D) Relative change in fat mass from baseline (% delta). Upward-pointing white triangles represent administration of exemplary anti-ALK7 antibody, downward-pointing white triangles represent administration of exemplary anti-GIP receptor antibody, and white squares represent administration of a combination of exemplary anti-ALK7 antibody and exemplary anti-GIP receptor antibody.

[0023] [Figure 6]Figures 6A-6C: A) Fasting blood glucose (mg / dL), B) Fasting plasma insulin (ng / mL), C) Liver volume, and fat and lean body mass composition (g). Upward-pointing white triangles represent administration of exemplary anti-ALK7 antibody, downward-pointing white triangles represent administration of exemplary anti-GIP receptor antibody, and white squares represent administration of a combination of exemplary anti-ALK7 antibody and exemplary anti-GIP receptor antibody. [Figure 7] Figures 7A-7D: A) Relative change in body weight from baseline (% delta), B) Daily food intake (g), C) Relative change in fat mass from baseline (% delta), D) Relative change in fat mass from baseline (% delta). White circles represent controls, upward-pointing white triangles represent administration of exemplary anti-ALK7 antibody, downward-pointing white triangles represent administration of exemplary anti-ActRII antibody, and white squares represent administration of exemplary anti-ALK7 antibody and exemplary anti-ActRII antibody combination. [Figure 8] Figures 8A-8C: A) Fasting blood glucose (mg / dL), B) Fasting plasma insulin (ng / mL), C) Liver volume, and fat and lean body mass composition (g). White circles represent controls, upward-pointing white triangles represent administration of exemplary anti-ALK7 antibody, downward-pointing white triangles represent administration of exemplary anti-ActRII antibody, and white squares represent administration of exemplary anti-ALK7 antibody and exemplary anti-ActRII antibody combination. [Figure 9] Figures 9A-9D: A) Relative change in body weight from baseline (% delta), B) Daily food intake (g), C) Relative change in fat mass from baseline (% delta), D) Relative change in fat mass from baseline (% delta). White circles represent controls, upward-pointing white triangles represent administration of exemplary anti-ALK7 antibody, downward-pointing white triangles represent administration of exemplary GLP-1 receptor agonist, white squares represent administration of exemplary combinations of GLP-1 receptor agonist and anti-ALK7i antibody, white diamonds represent administration of exemplary GLP-1 receptor-amyrin receptor agonist, and black squares represent administration of exemplary anti-ALK7 antibody and exemplary GLP-1 receptor-amyrin receptor agonist combinations. [Figure 10]Figures 10A-10C: A) Fasting blood glucose (mg / dL), B) Fasting plasma insulin (ng / mL), C) Liver volume, and fat and lean body mass composition (g). White circles represent controls, upward-pointing white triangles represent administration of exemplary anti-ALK7 antibody, downward-pointing white triangles represent administration of exemplary GLP-1 receptor agonist, white squares represent administration of exemplary combinations of GLP-1 receptor agonist and anti-ALK7i antibody, white diamonds represent administration of exemplary GLP-1 receptor-amyrin receptor agonist, and black squares represent administration of exemplary anti-ALK7 antibody and exemplary GLP-1 receptor-amyrin receptor agonist combinations.

[0024] Several definitions In general, terms used herein follow their meanings as understood in the art unless otherwise explicitly indicated. Explicit definitions of specific terms are provided below. The meanings of these terms and other terms in specific examples throughout this specification will be evident to those skilled in the art from the context.

[0025] To make the present invention easier to understand, several terms are first defined below. Additional definitions of the following terms and other terms are provided throughout this specification.

[0026] Approximately: When used herein in reference to a value, the term "approximately" refers to a value similar to the reference value. Generally, those skilled in the art will be familiar with the context and will understand the degree of relevant variance encompassed by "approximately" in that context. For example, in some embodiments, the term "approximately" may encompass a range of values ​​of 25%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, or less than 1% of the reference value.

[0027] Affinity: As is known in the art, affinity is a measure of the tightness with which two or more binding partners associate with each other. Those skilled in the art will recognize the various assays that can be used to assess affinity and will also recognize appropriate controls for such assays. In some embodiments, affinity is assessed by quantitative assays. In some embodiments, affinity is assessed over multiple concentrations (e.g., one binding partner at a time). In some embodiments, affinity is assessed in the presence of one or more potential competing entities (e.g., relevant, which may be present in a physiological setting). In some embodiments, affinity is assessed against a reference, e.g., a reference with known affinity above a certain threshold (positive control reference), or a reference with known affinity below a certain threshold (negative control reference). In some embodiments, affinity may be assessed against a simultaneous reference, and in some embodiments, affinity may be assessed against a past reference. Typically, when affinity is assessed against a reference, it is assessed under equivalent conditions. Agonist: The term agonist may be used herein to refer to a ligand, agent, state, or event whose presence, level, degree, type, or form correlates with an increase in the level or activity of another agent (i.e., an agonizing agent or target agent). Generally, an agonist may be, or may include, any chemical class of agent, including, for example, polypeptides, small molecules, nucleic acids, carbohydrates, lipids, metals, and / or any other entities exhibiting related activating activity. In some embodiments, the agonist may be direct (in which case the agonist directly affects its target). In some embodiments, the agonist may be indirect (in which case the agonist exerts an effect other than binding to its target, for example, by interacting with modifiers of the target, thereby changing the level or activity of the target).

[0028] Amino Acids: In its broadest sense, as used herein, the term amino acid refers to compounds and / or substances that can, can, can, or have been incorporated into a polypeptide chain, for example, through the formation of one or more peptide bonds. In some embodiments, amino acids have the general structure H2N-C(H)(R)-COOH. In some embodiments, amino acids are native amino acids. In some embodiments, amino acids are non-native amino acids, in some embodiments, amino acids are D-amino acids, and in some embodiments, amino acids are L-amino acids. "Standard amino acids" refers to any of the 20 standard L-amino acids commonly found in native peptides. "Non-standard amino acids" refers to any amino acid other than standard amino acids, whether synthetically prepared or obtained from natural sources. In some embodiments, amino acids containing carboxy-terminal amino acids and / or amino-terminal amino acids in a polypeptide may contain structural modifications compared to the general structure described above. For example, in some embodiments, amino acids may be configured to be modified by methylation, amidation, acetylation, pegylation, glycosylation, phosphorylation, and / or substitution (e.g., of an amino group, a carboxylic acid group, one or more protons, and / or a hydroxyl group) compared to their general structure. In some embodiments, such modifications may alter, for example, the cyclic half-life of a polypeptide containing a modified amino acid compared to a polypeptide otherwise containing the same unmodified amino acid. In some embodiments, such modifications do not significantly alter the related activity of a polypeptide containing a modified amino acid compared to a polypeptide otherwise containing the same unmodified amino acid. As is evident from the context, in some embodiments, the term amino acid may be used to refer to a free amino acid, and in some embodiments, it may be used to refer to an amino acid residue of a polypeptide.

[0029] Amylin: In this specification, the term "amylin" refers to a polypeptide having the same amino acid sequence as endogenous amylin, such as human amylin.

[0030] Amyrin receptor agonists: As used herein, “amyrin receptor agonists” refers to chemical entities characterized by being able to bind to and activate amyrin receptors. In some embodiments, “amyrin receptor agonists” are characterized by being able to bind to and activate at least the amyrin receptor 3 (AMYR3) complex. In some embodiments, amyrin receptor agonists are characterized by being able to stimulate calcitonin receptors and AMYR1-2.

[0031] Antagonists: Those skilled in the art will understand that, as used herein, the term “antagonist” can be used to refer to any agent, state, or event whose presence, level, degree, type, or form correlates with a reduction in the level or activity of another agent (i.e., an inhibitor, or target). Generally, an antagonist may be, or may include, any chemical class of agent, including, for example, small molecules, polypeptides, nucleic acids, carbohydrates, lipids, metals, and / or any other entities exhibiting related inhibitory activity. In some embodiments, the antagonist may be direct (in which case the antagonist directly affects its target). In some embodiments, the antagonist may be indirect (in which case the antagonist affects its level or activity of the target in a manner other than binding to the target, for example, by interacting with modifiers of the target).

[0032] Antibody: As used herein, the term antibody refers to a polypeptide containing standard immunoglobulin sequence elements sufficient to confer specific binding to a particular target antigen. As is known in the art, naturally produced intact antibodies are tetramers of about 150 kD, consisting of two identical heavy-chain polypeptides (each about 50 kD) and two identical light-chain polypeptides (each about 25 kD), which associate with each other in a structure commonly referred to as a Y-shaped structure. Each heavy chain contains at least four domains (each about 110 amino acids long), namely an amino-terminal variable (VH) domain (located at the tip of the Y structure), followed by three constant domains: CH1, CH2, and carboxy-terminal CH3 (located at the base of the stem of the Y). A short region known as the “switch” connects the heavy-chain variable and constant regions. The “hinge” connects the CH2 and CH3 domains to the rest of the antibody. Two disulfide bonds in this hinge region connect the two heavy-chain polypeptides to each other in an intact antibody. Each light chain consists of two domains: an amino-terminal variable (VL) domain followed by a carboxy-terminal constant (CL) domain, separated from each other by another switch. An intact antibody tetramer consists of two heavy-light dimers, where the heavy and light chains are linked to each other by a single disulfide bond; two other disulfide bonds link the heavy-chain hinge regions to each other, and the dimers link to each other to form a tetramer. Naturally produced antibodies are also typically glycosylated on the CH2 domain. Each domain of a natural antibody has a structure characterized by an immunoglobulin fold, formed from two beta sheets (e.g., 3, 4, or 5 strand sheets) packed together in a compressed antiparallel beta barrel. Each variable domain contains three hypervariable loops known as complement-determining regions (CDR1, CDR2, and CDR3) and four somewhat invariant framework regions (FR1, FR2, FR3, and FR4).When a natural antibody folds, the FR region forms a beta sheet that provides a structural framework for the domain, and the CDR loop regions from both the heavy and light chains are brought together in three-dimensional space to produce a single hypervariable antigen-binding site located at the tip of the Y structure. The Fc region of naturally occurring antibodies binds to elements of the complement system and also to receptors on effector cells, including effector cells that mediate cytotoxicity, for example. As is known in the art, the affinity and / or other binding properties of the Fc region to Fc receptors can be modulated through glycosylation or other modifications. In some embodiments, antibodies produced and / or utilized according to the present invention include a glycosylated Fc domain, which includes an Fc domain having such glycosylation that has been modified or manipulated. For the purposes of the present invention, in certain embodiments, any polypeptide or polypeptide complex containing a sufficient immunoglobulin domain sequence found in a natural antibody may be designated as an antibody and / or used as an antibody, whether the polypeptide is naturally produced (e.g., by an organism that reacts to an antigen) or produced by recombinant engineering, chemical synthesis, or other artificial systems or methodologies. In some embodiments, the antibody is polyclonal, and in some embodiments, the antibody is monoclonal. In some embodiments, the antibody has a constant region sequence characteristic of mouse, rabbit, primate, or human antibodies. In some embodiments, the antibody sequence element is humanized, primated, chimeric, etc., as known in the art. Furthermore, as used herein, the term antibody may refer to any construct or form known or developed in the art for utilizing the structural and functional features of an antibody in an alternative presentation, in a suitable embodiment (unless otherwise stated or evident from the context).For example, in some embodiments, the antibodies used according to the present invention are selected forms from among intact IgA, IgG, IgE, or IgM antibodies; bispecific or multispecific antibodies; antibody fragments such as Fab fragments, Fab' fragments, F(ab')2 fragments, Fd' fragments, Fd fragments, and isolated CDRs or sets thereof; single-chain Fv; polypeptide-Fc fusions; single-domain antibodies, alternative scaffolds, or antibody mimetic bodies (e.g., anti-kalin, FN3 monobodies, DARPins, Affibodies, Affilins, Affimers, Affitins, Alphabodies, Avimer, Fynomers, Im7VLR, VNAR, Trimab, CrossMab, Trident); nanobodies, bi-nanobody (binanobodies), F(ab')2, Fab', di-sdFv, single-domain antibodies, trifunctional antibodies, diabodies, minibodies, etc. In some embodiments, the antibody may lack covalent modifications (e.g., glycan attachment) that it would have if produced in nature. In some embodiments, the antibody may contain covalent modifications (e.g., glycan, payload [e.g., detectable portion, therapeutic portion, catalytic portion, etc.], or other pendant groups [e.g., polyethylene glycol, etc.]).

[0033] The antigen-binding fragment: As used herein, “the antigen-binding fragment” means that a portion of the antibody or antibody agent described herein is bound to an antigen or a portion thereof. The antigen-binding fragment may be produced by any means. For example, in some embodiments, the antigen-binding fragment may be produced enzymatically or chemically by fragmentation of an intact antibody or antibody agent. Alternatively, in some embodiments, the antigen-binding fragment may be produced recombinantly (i.e., by the expression of an engineered nucleic acid sequence). In some embodiments, the antigen-binding fragment may be produced entirely or partially synthetically. In some embodiments, the antigen-binding antibody fragment may have an amino acid length of at least about 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, or 200 or more amino acids.

[0034] Combination therapy: As used herein, the term “combination therapy” refers to a clinical intervention in which a subject is exposed to two or more treatment regimens (e.g., two or more therapeutic agents or modalities), for example, simultaneously. “Combination therapy” is used herein interchangeably with “therapeutic combination.” In some embodiments, two or more treatment regimens may be administered simultaneously. In some embodiments, two or more treatment regimens may be administered sequentially (e.g., a first regimen administered before any dose or episode of a second regimen). In some embodiments, two or more treatment regimens may be administered in overlapping dosing regimens. In some embodiments, the administration of combination therapy may involve the administration of one or more therapeutic agents or treatments to a subject receiving one or more other agents or treatments. In some embodiments, combination therapy does not necessarily require that the individual agents be administered together in a single composition (or even need to be administered simultaneously). In some embodiments, two or more therapeutic agents or treatments of combination therapy are administered separately to the subject, for example, in separate compositions, via separate routes of administration (e.g., one drug orally and another intravenously), and / or at different times.

[0035] Complementarity-Determining Regions: The term "CDR" refers to the complementarity-determining region within an immunoglobulin variable region sequence. Each of the heavy and light chain variable regions has three CDRs, named CDR1, CDR2, and CDR3, respectively. The term "CDR set" refers to a group of three CDRs occurring in a single variable region capable of binding to an antigen. The precise boundaries of these CDRs have been defined differently according to different systems. The system described by Kabat (Kabat et al. (1987 and (1991)) provides not only an unambiguous residue numbering system applicable to any variable region of an antibody, but also precise residue boundaries defining the three CDRs. These CDRs may also be called Kabat CDRs. Chothia and Coworkers (Chothia and Lesk (1987) J.Mol.Biol.196:901-917, Chothia et al. (1989) Nature 342:877-883) found that despite significant diversity at the amino acid sequence level, certain sub-parts within Kabat CDRs adopt nearly identical peptide skeletal structures. These sub-parts are designated as L1, L2, and L3 or H1, H2, and H3, where "L" and "H" represent the light chain and heavy chain regions, respectively. These regions can be referred to as Chothia CDRs, which have boundaries overlapping with Kabat CDRs. For other boundaries defining CDRs that overlap with Kabat CDRs, see Padlan (1995) FASEB J.9:133-139 and MacCallum (1996) J.Mol. Biol. 262(5):732-45). Furthermore, other CDR boundary definitions do not have to strictly adhere to one of the systems herein, but may nevertheless be shortened or extended in consideration of predictions or experimental findings that certain residues or groups of residues, or even the entire CDR, do not significantly affect antigen binding, but will overlap with Kabat CDRs. The methods used herein may utilize CDRs defined according to any of these systems, but certain embodiments use CDRs defined by Kabat or Chothia.

[0036] Identity: As used herein, the term identity refers to the overall relationship between polynucleotide molecules (e.g., DNA molecules and / or RNA molecules) and / or polypeptide molecules. In some embodiments, polynucleotide molecules (e.g., DNA molecules and / or RNA molecules) and / or polypeptide molecules are considered substantially identical to one another if their sequences are identical by at least 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99%. The calculation of the identity percentage of two nucleic acid or polypeptide sequences can be carried out, for example, by aligning the two sequences for optimal comparison purposes (e.g., gaps can be introduced in one or both of the first and second sequences for optimal alignment, and non-identical sequences can be ignored for comparison purposes). In certain embodiments, the length of sequences aligned for comparison purposes is at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or substantially 100% of the length of the reference sequence. Next, the nucleotides at corresponding positions are compared. If a position in the first sequence is occupied by the same residue (e.g., a nucleotide or amino acid) as the corresponding position in the second sequence, then the molecules are identical at that position. The percentage of identity between two sequences is a function of the number of identical positions shared by the sequences, taking into account the number of gaps that need to be introduced for optimal alignment of the two sequences and the length of each gap. Sequence comparison and determination of the percentage of identity between two sequences can be achieved using mathematical algorithms. For example, the percentage of identity between two nucleotide sequences can be determined using the algorithm of Meyers and Miller, 1989, which is incorporated into the ALIGN program (version 2.0).In some exemplary embodiments, nucleic acid sequence comparisons performed using the ALIGN program utilize the PAM120 weight residue table, gap length penalty 12, and gap penalty 4. Alternatively, the identity percentage between two nucleotide sequences can also be determined using the GAP program within the GCG software package, using the NWSgapdna.CMP matrix.

[0037] GLP-1: In this specification, the terms "GL-1" or "natural GLP-1" refer to human glucagon-like peptide-1 (GLP-1(7-37)), which is listed as Sequence ID No. 7 in the sequence listing. In the sequence listing, amino acid residues are numbered sequentially from 1 to 31. Therefore, the first amino acid in wild-type human GLP-1(7-37), i.e., the N-terminal histidine, is number "1" in Sequence ID No. 7.

[0038] GLP-1 receptor agonists: In this disclosure, “GLP-1 receptor agonists” may be used as ligands that bind to the GLP-1 receptor and can produce a biological response similar to that of the endogenous ligand, glucagon-like peptide 1 (GLP-1). A “complete” GLP-1 receptor agonist may be defined as a GLP-1 receptor agonist that can induce a biological response similar to that of GLP-1.

[0039] Pharmaceutical Composition: As used herein, the term pharmaceutical composition refers to a composition suitable for administration to human or animal subjects. In some embodiments, the composition comprises one or more activators formulated with one or more pharmaceutically acceptable carriers. In some embodiments, the activators are present in unit doses suitable for administration in a therapeutic regimen. In some embodiments, the therapeutic regimen comprises one or more doses administered according to a schedule determined to exhibit a statistically significant probability of achieving the desired therapeutic effect when administered to subjects or populations in need of treatment. In some embodiments, the pharmaceutical composition may be specifically formulated for administration in solid or liquid form, including: oral administration, e.g., starch (aqueous or non-aqueous solution or suspension), tablets, e.g., tablets, bolus, powder, granules, paste for application to the tongue; parenteral administration, e.g., subcutaneous, intramuscular, intravenous, or epidural injection of sterile solution or suspension, or parenteral administration by sustained-release formulation; topical application, e.g., topical application as a cream, ointment, or sustained-release patch or spray applied to the skin, lungs, or oral cavity; intravaginal or rectal application, e.g., intravaginal or rectal, sublingual, ocular, transdermal, or intranasal, lungs, and other mucosal surfaces as an emulsion, cream, or foam. In some embodiments, the pharmaceutical composition is sterile and substantially free of pyrogens.

[0040] Pharmacopoeia-acceptable: As used herein, the term “pharmacopoeia-acceptable” means a compound, material, composition, and / or dosage form that, within the bounds of sound medical judgment, is suitable for use in contact with human and animal tissues to an extent commensurate with a reasonable benefit-to-risk ratio, without excessive toxicity, irritation, allergic reactions, or other problems or complications.

[0041] Treatment: As used herein, the term “treatment” refers to any medical therapy for any human or other vertebrate subject that requires it. The subject is expected to have undergone a physical examination by a physician or veterinarian who has made a provisional or definitive diagnosis indicating that the use of such particular treatment would be beneficial to the health of the human or other vertebrate. The timing and purpose of such treatment may vary from individual to individual, depending on the subject’s current health status. Therefore, such treatment may be prophylactic (preventive), palliative, symptomatic, and / or curative. [Modes for carrying out the invention]

[0042] This disclosure provides combinations such as combination therapies and / or compositions comprising (b) an anti-ALK7 GLP-1 receptor agonist, (b) an anti-GIP receptor antibody or its antigen-binding fragment, and / or (c) a second bioactive agent comprising an anti-Act receptor II antibody or its antigen-binding fragment. This disclosure also provides combinations, e.g., combination therapies and / or compositions described herein, for use in the prevention and / or treatment of diseases and / or disorders. In particular, this disclosure provides an anti-ALK7 receptor antibody or its antigen-binding fragment for use in combination with a second bioactive agent in the prevention and / or treatment of overweight, obesity, and / or diabetes.

[0043] Anti-ALK7 receptor antibody or its antigen-binding fragment Activin-like kinase 7 (ALK7) is a type I serine / threonine kinase that forms a hexameric signaling complex on its ligand-bound site. This signaling complex is a heterooligomer consisting of two type I and two type II transmembrane serine / threonine kinases. The type II receptor phosphorylates and activates the type I receptor, which then autophosphorylates and subsequently binds to and activates the SMAD transcription regulators, SMAD2 and SMAD3. ALK7 is the receptor for activin B. In adipose tissue, ALK7-mediated ActB signaling suppresses lipolysis, downregulates adrenergic receptors, and promotes fat accumulation, while inhibition of ALK7 has been shown to reduce body weight and have other metabolic benefits in preclinical animal models.

[0044] In some embodiments, an anti-ALK7 receptor antibody or its antigen-binding fragment specifically binds to the ALK7 receptor. In some embodiments, an anti-ALK7 receptor antibody or its antigen-binding fragment blocks the ALK7 receptor. In some embodiments, an anti-ALK7 receptor antibody or its antigen-binding fragment is an anti-ALK7 receptor antibody blocking antibody or its antigen-binding fragment. In some embodiments, an anti-ALK7 receptor antibody or its antigen-binding fragment is an ALK7 receptor antagonist, i.e., inhibits or blocks the ALK7 receptor. In some embodiments, an anti-ALK7 antibody receptor or its antigen-binding fragment antagonizes the ALK7 receptor. In some embodiments, an anti-ALK7 antibody receptor or its antigen-binding fragment inhibits ALK7 signaling. In some embodiments, an anti-ALK7 receptor antibody or its antigen-binding fragment binds to the ALK7 receptor and inhibits its biological activity, such as the ability to suppress lipolysis and / or prevent fat accumulation. In some embodiments, an anti-ALK7 receptor antibody or its antigen-binding fragment increases lipolysis. In some embodiments, the anti-ALK7 receptor antibody or its antigen-binding fragment blocks the formation of the hexameric signaling complex of the ALK7 receptor having an ActRIIA / B type II receptor. In some embodiments, the anti-ALK7 receptor antibody or its antigen-binding fragment inhibits the activation of SMAD2 and / or SMAD3. In some embodiments, the anti-ALK7 receptor antibody or its antigen-binding fragment blocks the binding of ALK7 receptor ligands such as activin B, GDF11, activin A, BMP10, BMP6, BMP5, GDF3, activin C, activin E, activin AC, activin BC, activin AE, node, or activin BE. In some embodiments, the anti-ALK7 receptor antibody or its antigen-binding fragment blocks the binding of activin B to the ALK7 receptor. In some embodiments, the anti-ALK7 receptor antibody or its antigen-binding fragment blocks the formation of the hexameric signaling complex of the ALK7 receptor having an ActRIIA / B type II receptor.

[0045] The human ALK7 receptor gene (uniprotkb / Q8NER5) encodes a protein consisting of 493 amino acid residues, including a signal peptide, extracellular domain, transmembrane domain, and cytoplasmic domain. The ALK7 receptor protein sequence is MTRALCSALRQALLLLAAAAELSPGLKCVCLLCDSSNFTCQTEGACWASVMLTNGKEQVIKSCVSLPELNAQVFCHSSNNVTKTECCFTDFCNNITLHLPTASPNAPKLGPMELAIIITVPVCLLSIAAMLTVWACQGRQCSYRKKKRPNVEEPLSECNLVNAGKTLKDLIYDVTASGSGSGLPLLVQRTIARTIVLQEIVGKGRFGEVWHGRWCGEDVAVKIFSSRDERSWFREAEIYQTVML RHENILGFIAADNKDNGTWTQLWLVSEYHEQGSLYDYLNRNIVTVAGMIKLALSIASGLAHLHMEIVGTQGKPAIAHRDIKSKNILVKKCETCAIADLGLAVKHDSILNTIDIPQNPKVGTKRYMAPEMLDDTMNVNIFESFKRADIYSVGLVYWEIARRCSVGGIVEEYQLPYYDMVPSDPSIEEMRKVVCDQKFRPSIPNQWQSCEALRVMGRIMRECWYANGAARLTALRIKKTISQLCVKEDCKA (sequence number 1). In some embodiments, an anti-ALK7 receptor antibody or its antigen-binding fragment binds to the extracellular domain of the ALK7 receptor, which includes amino acid residues 26-113 of SEQ ID NO: 1, or a portion thereof. In some embodiments, the extracellular domain (ECD) of the ALK7 receptor extends from the signal peptide to the transmembrane region, i.e., to the amino acid residues at positions 26-113 of the human ALK7 receptor coding region as defined by SEQ ID NO: 1. In some embodiments, the ECD of truncated ALK7 is truncated at the N-terminus to include only amino acids 26-113, and is referred to herein as ALK7-ECD26-113. In some embodiments, an anti-ALK7 receptor antibody or its antigen-binding fragment binds to human ALK7-ECD26-113.

[0046] In some embodiments, the anti-ALK7 receptor antibody or its antigen-binding fragment includes two variable regions. In some embodiments, the anti-ALK7 receptor antibody or its antigen-binding fragment includes an antigen-binding domain comprising six CDR sequences. In some embodiments, the anti-ALK7 receptor antibody or its antigen-binding fragment includes two variable regions, each comprising three CDR sequences. In some embodiments, the variable regions include several framework sequences. In some embodiments, the variable region sequences include, for example, FR1-CDR1-FR2-CDR2-FR3-CDR3-FR4 as defined by Kabat, Chothia, and / or IMGT.

[0047] In some embodiments, the CDR is defined according to Kabat and / or IMGT. In some embodiments, the CDR is defined according to Kabat. In some embodiments, the CDR is defined according to IMGT. In some embodiments, the CDR is defined according to Chothia.

[0048] In some embodiments, the anti-ALK7 receptor antibody or its antigen-binding fragment comprises a heavy chain and a light chain. In some embodiments, the anti-ALK7 receptor antibody or its antigen-binding fragment comprises a variable heavy chain sequence and a variable light chain sequence. In some embodiments, the anti-ALK7 receptor antibody or its antigen-binding fragment comprises a heavy chain and a light chain variable region, each sequence comprising three CDRs. In some embodiments, the anti-ALK7 receptor antibody or its antigen-binding fragment comprises a heavy chain variable sequence comprising three CDRs and a light chain variable sequence comprising three CDRs.

[0049] An exemplary amino acid sequence of an anti-ALK7 antibody is shown in Table A below. [Table 1] JPEG2026167601000006.jpg193170 JPEG2026167601000007.jpg161170

[0050] In some embodiments, the present disclosure provides an anti-ALK7 receptor antibody or its antigen-binding fragment that binds to the extracellular domain of the human ALK7 receptor (SEQ ID NO: 1) and comprises three heavy chain CDR sequences hCDR1-3 and three light chain CDR sequences lCDR1-3. a) hCDR1 is SGNYWN (SEQ ID NO: 8) which has a maximum of one or two amino acid substitutions. b) hCDR2 is YISFDGRNNYNPSLKN (SEQ ID NO: 10) having up to 1, 2, or 3 amino acid substitutions, c)hCDR3 is DYYGSEGFGY (SEQ ID NO: 17) which has a maximum of 1, 2, or 3 amino acid substitutions. d) lCDR1 is KSSQSLLNSRTRKNYLA (SEQ ID NO: 19) having up to 1, 2, or 3 amino acid substitutions, e) lCDR2 is WTSTRES (SEQ ID NO: 22) with a maximum of one or two amino acid substitutions, f) ICDR3 is KQSYNLPYT (SEQ ID NO: 24) which has a maximum of one or two amino acid substitutions.

[0051] In some embodiments, the present disclosure provides antibodies such as anti-ALK7 receptor antibodies or antigen-binding fragments thereof that bind to the extracellular domain of the human ALK7 receptor (SEQ ID NO: 1) and comprise three heavy chain CDR sequences hCDR1-3 and three light chain CDR sequences lCDR1-3. a) hCDR1 is SGSYWN (SEQ ID NO: 9) having a maximum of one or two amino acid substitutions, b) hCDR2 is YISFDGRTNYNPSLKN (SEQ ID NO: 11) having up to 1, 2, or 3 amino acid substitutions, c)hCDR3 is DYYGSEGFAY (SEQ ID NO: 18) which has up to 1, 2, or 3 amino acid substitutions. d) lCDR1 is KSSQSLLVSRTRKNYLA (SEQ ID NO: 20) having up to 1, 2, or 3 amino acid substitutions, e) lCDR2 is WTSTRES (SEQ ID NO: 22) with a maximum of one or two amino acid substitutions, f) ICDR3 is KQSYNLPYT (SEQ ID NO: 24) which has a maximum of one or two amino acid substitutions.

[0052] In some embodiments, the present disclosure provides antibodies such as anti-ALK7 receptor antibodies or antigen-binding fragments thereof that bind to the extracellular domain of the human ALK7 receptor (SEQ ID NO: 1) and comprise three heavy chain CDR sequences hCDR1-3 and three light chain CDR sequences lCDR1-3. a) hCDR1 is SGSYWN (sequence number 9), b) hCDR2 is YISFDGRTNYNPSLKN (Sequence ID 11), c)hCDR3 is DYYGSEGFAY (Sequence ID 18), d) lCDR1 is KSSQSLLVSRTRKNYLA (Sequence ID 20), e) lCDR2 is WTSTRES (sequence number 22), f) lCDR3 is KQSYNLPYT (sequence number 24).

[0053] In some embodiments, the present disclosure provides antibodies such as anti-ALK7 receptor antibodies or antigen-binding fragments thereof that bind to the extracellular domain of the human ALK7 receptor (SEQ ID NO: 1) and comprise three heavy chain CDR sequences hCDR1-3 and three light chain CDR sequences lCDR1-3. a) hCDR1 is SGNYWN (SEQ ID NO: 8) or SGSYWN (SEQ ID NO: 9) which has a maximum of one amino acid substitution. b) hCDR2 is YISFDGRNNYNPSLKN (SEQ ID NO: 10), YISFDGRTNYNPSLKN (SEQ ID NO: 11), YISFDGRTNYNPSLKS (SEQ ID NO: 12), YISFDGRRNYNPSLKN (SEQ ID NO: 13), YISFTGRTNYNPSLKN (SEQ ID NO: 14), YISFSGRNNYNPSLKN (SEQ ID NO: 15), or YISFSGRTNYNPSLKN (SEQ ID NO: 16), which have a maximum of one or two amino acid substitutions. c)hCDR3 is DYYGSEGFGY (SEQ ID NO: 17) or DYYGSEGFAY (SEQ ID NO: 18) which has a maximum of one or two amino acid substitutions. d) lCDR1 is KSSQSLLNSRTRKNYLA (SEQ ID NO: 19), KSSQSLLVSRTRKNYLA (SEQ ID NO: 20), or KSSQSLLYSRTRKNYLA (SEQ ID NO: 21), having up to one or two amino acid substitutions. e) lCDR2 is WTSTRES (SEQ ID NO: 22) or WASTRES (SEQ ID NO: 23) which has a maximum of one amino acid substitution. f) ICDR3 is KQSYNLPYT (SEQ ID NO: 24) or KRSYNLPYT (SEQ ID NO: 25) with a maximum of one amino acid substitution.

[0054] In some embodiments, the amino acid substitution is a conservative substitution or an alanine substitution.

[0055] In some embodiments, the present disclosure provides antibodies such as anti-ALK7 receptor antibodies or antigen-binding fragments thereof that bind to the extracellular domain of the human ALK7 receptor (SEQ ID NO: 1) and comprise three heavy chain CDR sequences hCDR1-3 and three light chain CDR sequences lCDR1-3. a) hCDR1 is SGNYWN (sequence number 8) or SGSYWN (sequence number 9), b) hCDR2 is YISFDGRNNYNPSLKN (SEQ ID NO: 10), YISFDGRTNYNPSLKN (SEQ ID NO: 11), YISFDGRTNYNPSLKS (SEQ ID NO: 12), YISFDGRRNYNPSLKN (SEQ ID NO: 13), YISFTGRTNYNPSLKN (SEQ ID NO: 14), YISFSGRNNYNPSLKN (SEQ ID NO: 15), or YISFSGRTNYNPSLKN (SEQ ID NO: 16), c)hCDR3 is DYYGSEGFGY (Sequence ID 17) or DYYGSEGFAY (Sequence ID 18), d) lCDR1 is KSSQSLLNSRTRKNYLA (sequence number 19), KSSQSLLVSRTRKNYLA (sequence number 20), or KSSQSLLYSRTRKNYLA (sequence number 21), e) lCDR2 is WTSTRES (sequence number 22) or WASTRES (sequence number 23), f) lCDR3 is either KQSYNLPYT (sequence number 24) or KRSYNLPYT (sequence number 25).

[0056] In some embodiments, the present disclosure provides antibodies such as anti-ALK7 receptor antibodies or antigen-binding fragments thereof that bind to the extracellular domain of the human ALK7 receptor (SEQ ID NO: 1) and comprise three heavy chain CDR sequences hCDR1-3 and three light chain CDR sequences lCDR1-3, where, i. The three heavy chain CDR sequences are selected from any one of the heavy chain variable regions (HVs) identified by SEQ ID NOs. 26-59, and / or ii. The three light chain CDR sequences are selected from any one of the light chain variable regions (LVs) identified by sequence numbers 60-84.

[0057] In some embodiments, the anti-ALK7 receptor antibody or its antigen-binding fragment comprises a heavy chain variable sequence, a light chain variable sequence, or both. In some embodiments, the anti-ALK7 antibody or its antigen-binding fragment comprises (a) A heavy chain variable region containing an amino acid sequence that is at least 70%, for example, at least 75%, for example, at least 80%, for example, at least 85%, for example, at least 90%, for example, at least 91%, for example, at least 92%, for example, at least 93%, for example, at least 94%, for example, at least 95%, for example, at least 96%, for example, at least 97%, for example, at least 98%, for example, at least 99%, for example, 100% identical to SEQ ID NO: 43, (b) A light chain variable region comprising an amino acid sequence that is at least 70%, for example, at least 75%, for example, at least 80%, for example, at least 85%, for example, at least 90%, for example, at least 91%, for example, at least 92%, for example, at least 93%, for example, at least 94%, for example, at least 95%, for example, at least 96%, for example, at least 97%, for example, at least 98%, for example, at least 99%, for example, 100% identical to SEQ ID NO: 78.

[0058] In some embodiments, the anti-ALK7 antibody or its antigen-binding fragment is (a) A heavy chain variable region containing the amino acid sequence according to Sequence ID No. 43, (b) A light chain variable region containing the amino acid sequence according to SEQ ID NO: 78, and

[0059] In some embodiments, the anti-ALK7 receptor antibody or its antigen-binding fragment includes a heavy chain constant region, a light chain constant region, or both. In some embodiments, the anti-ALK7 antibody or its antigen-binding fragment is (a) A heavy chain variable region containing an amino acid sequence that is at least 70%, for example, at least 75%, for example, at least 80%, for example, at least 85%, for example, at least 90%, for example, at least 91%, for example, at least 92%, for example, at least 93%, for example, at least 94%, for example, at least 95%, for example, at least 96%, for example, at least 97%, for example, at least 98%, for example, at least 99%, for example, 100% identical to SEQ ID NO: 85, (b) A light chain constant region comprising an amino acid sequence that is at least 70%, for example, at least 75%, for example, at least 80%, for example, at least 85%, for example, at least 90%, for example, at least 91%, for example, at least 92%, for example, at least 93%, for example, at least 94%, for example, at least 95%, for example, at least 96%, for example, at least 97%, for example, at least 98%, for example, at least 99%, for example, 100% identical to SEQ ID NO: 86.

[0060] In some embodiments, the anti-ALK7 antibody or its antigen-binding fragment is (a) A heavy chain variable region containing the amino acid sequence according to Sequence ID No. 85, (b) A light chain constant region comprising the amino acid sequence according to SEQ ID NO: 86, and

[0061] In some embodiments, the anti-ALK7 receptor antibody or its antigen-binding fragment comprises a heavy chain, a light chain, or both. In some embodiments, the anti-ALK7 antibody or its antigen-binding fragment is (a) A heavy chain containing an amino acid sequence that is at least 70%, for example, at least 75%, for example, at least 80%, for example, at least 85%, for example, at least 90%, for example, at least 91%, for example, at least 92%, for example, at least 93%, for example, at least 94%, for example, at least 95%, for example, at least 96%, for example, at least 97%, for example, at least 98%, for example, at least 99%, for example, 100% identical to SEQ ID NO: 99, (b) A light chain comprising an amino acid sequence that is at least 70%, for example, at least 75%, for example, at least 80%, for example, at least 85%, for example, at least 90%, for example, at least 91%, for example, at least 92%, for example, at least 93%, for example, at least 94%, for example, at least 95%, for example, at least 96%, for example, at least 97%, for example, at least 98%, for example, at least 99%, for example, 100% identical to SEQ ID NO: 100.

[0062] In some embodiments, the anti-ALK7 antibody or its antigen-binding fragment is (a) A heavy chain containing the amino acid sequence according to SEQ ID NO: 99, (b) A light chain variable region containing the amino acid sequence according to Sequence ID No. 100, and

[0063]

[0064] In some embodiments, the anti-ALK7 receptor antibody or its antigen-binding fragment is a heavy chain or comprises a heavy chain. In some embodiments, the heavy chain of the anti-ALK7 receptor antibody or its antigen-binding fragment is QVQLVQSGAEVKKPGSSVKVSCKASGGTFSGQAISWVRQAPGQGLEWMGGIIPSFGTARYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARTPYYDSSGYLDVWGQGTMVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPEAEGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHED An amino acid sequence having at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of PEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPSSIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK (Sequence ID 97), or containing such an amino acid sequence.

[0065] In some embodiments, the heavy chain of the anti-ALK7 receptor antibody or its antigen-binding fragment is QVQLQESGPGLVKPSESLSLTCTVEGYSITSGSYWNWIRQPPGKGLEWIGYISFDGRTNYNPSLKNRVTISRDTSKNQFSLKLSSVTAADTAVYYCARDYYGSEGFAYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPEAEGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDP An amino acid sequence having at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of EVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPSSIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK (SEQ ID NO: 99), or containing the same.

[0066] In some embodiments, the anti-ALK7 receptor antibody or its antigen-binding fragment is or comprises a light chain. In some embodiments, the light chain of the anti-ALK7 receptor antibody or its antigen-binding fragment is or comprises an amino acid sequence having at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of DIQMTQSPSSLSASVGDRVTITCQASHDIDNYLNWYQQKPGKAPKLLIYYASNLKTGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCQQSRASPPTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 98).

[0067] In some embodiments, the anti-ALK7 receptor antibody or its antigen-binding fragment is a light chain or comprises one. In some embodiments, the anti-ALK7 receptor antibody or its antigen-binding fragment light chain is an amino acid sequence having at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of DIVMTQSPDSLAVSLGEDATINCKSSQSLLVSRTRKNYLAWYQQKPGQPPKLLIYWTSTRESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCKQSYNLPYTFGGGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 100), or includes the same.

[0068] In some embodiments, the anti-ALK7 receptor antibody or its antigen-binding fragment is a Fab, Fab', F(ab')2, Fv, Fd, dAb, scFv, or single-domain antibody. In some embodiments, the anti-ALK7 receptor antibody or its antigen-binding fragment is a full-length antibody. In some embodiments, the anti-ALK7 receptor antibody or its antigen-binding fragment is an IgG antibody such as an IgG1, IgG2, IgG3, or IgG4 antibody. In some embodiments, the anti-ALK7 receptor antibody or its antigen-binding fragment includes heavy and light chain constant regions of an IgG subtype such as IgG1, IgG2, IgG3, or IgG4.

[0069] In some embodiments, the heavy chain constant region comprises the amino acid sequence of SEQ ID NO: 85 (the C-terminal lysine is optional). In some embodiments, the light chain constant region comprises SEQ ID NO: 86. In some embodiments, the anti-ALK7 receptor antibody or its antigen-binding fragment comprises a heavy chain constant region and a light chain constant region, each comprising one or more amino acid substitutions to optimize Fc function. This may include amino acid substitutions that affect cell interactions or effector-protein interactions, such as FcR effector function or complement system effector function, such as mutations equivalent to L117A, L118E, G120A, A213S, and P214S in SEQ ID NO: 85. Using the EU numbering system, the mutations are referred to as L234A, L235E, G237A, A330S, and / or P331S. In some embodiments, the constant region comprises one or more amino acid substitutions selected from the group consisting of L234A, L235E, G237A, A330S, P331S, or combinations thereof.

[0070] The Disclosure also provides a method for producing the anti-ALK7 antibody or its antigen-binding fragment provided herein. The method may comprise cloning such an anti-ALK7 antibody in a suitable expression vector and establishing a host cell that expresses the antibody. In some embodiments, the Disclosure also provides a nucleotide sequence or pair of nucleotide sequences encoding the anti-ALK7 antibody or its antigen-binding fragment provided herein. In some embodiments, the Disclosure also provides a host cell comprising a nucleotide sequence or pair of nucleotide sequences encoding the anti-ALK7 antibody or its antigen-binding fragment provided herein. In some embodiments, the Disclosure also provides a host cell that expresses the anti-ALK7 antibody or its antigen-binding fragment provided herein. In some embodiments, the Disclosure provides a method for producing the anti-ALK7 antibody or its antigen-binding fragment, a) Culturing host cells expressing the anti-ALK7 antibody or its antigen-binding fragment described herein, b) The present invention also provides a method comprising purifying the anti-ALK7 receptor antibody or its antigen-binding fragment.

[0071] A method for preparing exemplary anti-ALK7 receptor antibodies or their antigen-binding fragments is described in Example 2 of this specification.

[0072] The in vitro potency of an anti-ALK7 receptor antibody or its antigen-binding fragment may be measured as described in Example 3 of this specification. The potency of an anti-ALK7 receptor antibody or its antigen-binding fragment may also be described by its IC50 value, where IC50 is the concentration of the inhibitor (e.g., anti-ALK7 receptor antibody or its antigen-binding fragment) at which the response is halved. The lower the IC50 value, the more potent the compound. In some embodiments, the anti-ALK7 receptor antibody or its antigen-binding fragment described herein has an IC50 value of less than 5 nM, e.g., less than 4 nM, e.g., less than 3 nM, e.g., less than 2.5 nM, e.g., less than 2 nM, relative to the human ALK7 receptor.

[0073] The affinity of an anti-ALK7 receptor antibody or its antigen-binding fragment for that antigen can be determined by various methods, such as surface plasmon resonance (SPR), where the dynamics of complex formation and dissociation are measured. In some embodiments, the affinity (KD) for the anti-ALK7 antibody or its antigen-binding fragment is 1 to 1000 nM, e.g., 10 to 750 nM, or e.g., 10 to 500 nM. In some embodiments, the affinity (KD) for the anti-ALK7 antibody or its antigen-binding fragment is less than 10 μM, e.g., less than 5 μM, e.g., less than 1 μM. In some embodiments, the affinity (KD) for the anti-ALK7 antibody or its antigen-binding fragment is less than 100 nM, e.g., less than 50 nM, e.g., less than 10 nM.

[0074] Second bioactive agent This disclosure provides combinations, e.g., combination therapies, and / or compositions comprising an anti-ALK7 receptor antibody or its antigen-binding fragment and a second bioactive agent. This disclosure provides an anti-ALK7 receptor antibody or its antigen-binding fragment for use in the prevention and / or treatment of overweight, obesity, and / or diabetes in combination with a second bioactive agent.

[0075] In some embodiments, the second bioactive agent is one or more agonists, such as receptor agonists, or contains such agonists. In some embodiments, the second bioactive agent is one or more antagonists, such as receptor antagonists, or contains such agonists. In some embodiments, the second bioactive agent is one or more antibodies or antigen-binding fragments thereof, or contains such agonists. In some embodiments, the second bioactive agent is an incretin hormone or a variant thereof.

[0076] In some embodiments, the second bioactive agent is or comprises a GLP-1 receptor agonist.

[0077] In some embodiments, the second bioactive agent is or comprises a GLP-1 receptor-amyrin receptor agonist.

[0078] In some embodiments, the second bioactive agent is a GIP receptor antagonist, such as an anti-GIP receptor antibody or an antigen-binding fragment thereof, or contains the same. In some embodiments, the second bioactive agent is an ActRII antagonist, such as an anti-ActRII antibody or an antigen-binding fragment thereof, or contains the same.

[0079] GLP-1 receptor agonists Glucagon-like peptide 1 (GLP-1) is a hormone derived from enteroendocrine cells and is one of two prominent endogenous physiological incretins. GLP-1 improves blood glucose control by stimulating glucose-dependent insulin secretion in response to nutrients (glucose), inhibits glucagon secretion from pancreatic alpha cells, slows gastric emptying, and induces weight loss primarily by reducing food consumption.

[0080] In some embodiments, the second bioactive agent activates the GLP-1 receptor. In some embodiments, the second bioactive agent is or comprises a GLP-1 receptor agonist, i.e., a GLP-1 analog. In some embodiments, a combination, e.g., combination therapy, and / or composition comprises an anti-ALK7 receptor antibody or its antigen-binding fragment, and a GLP-1 receptor agonist.

[0081] In some embodiments, the present disclosure provides an anti-ALK7 receptor antibody or its antigen-binding fragment for use in combination with a GLP-1 receptor agonist in the prevention and / or treatment of overweight, obesity, and / or diabetes.

[0082] In some embodiments, GLP-1 receptor agonists are characterized by binding to and activating or activating the GLP-1 receptor. In other words, in some embodiments, GLP-1 receptor agonists have "potency" for the GLP-1 receptor. The in vitro potency of a GLP-1 receptor agonist for the GLP-1 receptor can be measured as described in Assay 1 of WO / 2022 / 129526. The potency of a GLP-1 receptor agonist can be described by its EC50 value, which represents the concentration of the compound at which 50% of its maximum effect is observed. A lower EC50 value indicates a more potent compound. In some embodiments, the GLP-1 receptor agonists disclosed herein have an EC50 value of less than 300 pM, for example, less than 200 pM, for example, less than 150 pM, preferably less than 100 pM, for example less than 75 pM, and more preferably less than 50 pM, for example less than 40 pM, for example less than 30 pM, for example less than 20 pM, for example less than 13 pM, relative to the human GLP-1 receptor.

[0083] Examples of exogenous GLP-1 receptor agonists include semaglutide (the active ingredient in Ozempic®, Rybelsus®, and Wegovy®), liraglutide (the active ingredient in Victoza® and Saxenda®), tylzepatide (the active ingredient in Mounjaro® and Zepbound®), and dulaglutide (the active ingredient in Trulicity®).

[0084] Exemplary GLP-1 agonists are provided in WO2006 / 097537 and WO / 2022 / 129526, which are incorporated herein by reference.

[0085] In some embodiments, the GLP-1 receptor agonist comprises a non-natural amino acid. In some embodiments, the non-natural amino acid is alpha(α)-aminoisobutyryl, α-aminoisobutyric acid, or α-methylalanine, abbreviated herein as Aib. In some embodiments, alpha-aminoisobutyryl is located at the N-terminal portion of the GLP-1 sequence. In some embodiments, the GLP-1 receptor agonist is an amino acid sequence having at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of H-Aib-EGTFTSDVSSYLEGQAAKEFIAWLVRGRG (SEQ ID NO: 2), or comprises such an amino acid sequence. As previously stated, Aib is 2-aminoisobutyric acid.

[0086] In some embodiments, the GLP-1 receptor agonist includes one to three extensions. In some embodiments, the extension is covalently bonded to the lysine (K) residue at position 20 of SEQ ID NO: 2. In some embodiments, the GLP-1 receptor agonist includes a fatty acid extension factor. In some embodiments, the fatty acid extension factor is a C16-C20 diacid, for example, a C18 diacid.

[0087] In some embodiments, the GLP-1 receptor agonist comprises or is represented by the following formula I (SEQ ID NO: 106): [ka] (Formula I, Sequence ID 106), or a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable salt thereof.

[0088] In some embodiments, the GLP-1 receptor agonist is or comprises semaglutide (an exemplary GLP-1 receptor agonist). In some embodiments, the GLP-1 receptor agonist is N6.26-{18-[N-(17-carboxyheptadecanoyl)-L-γ-glutamyl]-10-oxo-3,6,12,15-tetraoxa-9,18-diazaoctadecanoyl}-[8-(2-amino-2-propanoic acid),34-L-arginine]human glucagon-like peptide 1(7-37). Semaglutide can be prepared using methods well known to those skilled in the art, such as those described in WO2006 / 097537 and J.Med. Chem.2015,58,18,7370-7380, and the method briefly described in Example 4 of WO2006 / 097537.

[0089] In some embodiments, the GLP-1 receptor agonist comprises or conforms to the following formula II (SEQ ID NO: 107). [ka] (Formula II, Sequence ID No. 107) or a pharmaceutically acceptable salt thereof.

[0090] In some embodiments, the semaglutide exists in its fully or partially ionized form. In some embodiments, one or more carboxylic acid groups (-COOH) of the semaglutide may be deprotonated to a carboxylate group (-COO-), and / or one or more amino groups (-NH2) may be protonated to an -NH3+ group. In some embodiments, the semaglutide may be in the form of a salt, preferably a pharmaceutically acceptable salt.

[0091] GLP-1 receptor-amyrin receptor agonists In some embodiments, the second bioactive agent is or contains an amyrin receptor agonist, such as an amyrin receptor 3 agonist. In some embodiments, the second bioactive agent contains two or more agonists. In some embodiments, the second bioactive agent is or contains a GLP-1 receptor agonist and an amyrin receptor agonist, such as an amyrin receptor 3 agonist. In some embodiments, the second bioactive agent is or contains a GLP-1 receptor-amyrin receptor agonist. In some embodiments, the GLP-1 receptor-amyrin receptor agonist is a single molecule.

[0092] In some embodiments, amyrin receptor agonists activate and / or activate the calcitonin receptor (CTR) and / or the amyrin receptor (AMYR). The amyrin receptor consists of a heterodimer of two components, namely, the calcitonin receptor (CTR) and one of three receptor activity-modifying proteins (RAMP1-3), resulting in three possible complexes AMYR1-3. Unless otherwise specified herein, “amyrin receptor” refers to at least amyrin receptor 3 (AMYR3). In some embodiments, a second bioactive agent containing an amyrin receptor agonist has cross-reactivity to both the amyrin receptor and the calcitonin receptor. In some embodiments, a second bioactive agent containing an amyrin receptor agonist activates and / or activates the amyrin receptor but does not have cross-reactivity to the calcitonin receptor. In some embodiments, the second bioactive agent described herein contains an amyrin receptor agonist. Examples of endogenous amyrin receptor agonists are human amyrin and human calcitonin. Examples of exogenous amyrin receptor agonists include pramulintide and caglilintide, as disclosed in WO2012 / 168432, which is incorporated herein by reference.

[0093] In some embodiments, the GLP-1 receptor-amyrin receptor agonist is a peptide or comprises a peptide. In some embodiments, the GLP-1 receptor-amyrin receptor agonist is a single-chain polypeptide skeleton comprising one, two, or three lysine (Lys,K) residues or comprises a single-chain polypeptide skeleton. In some embodiments, the GLP-1 receptor-amyrin receptor agonist is a single-chain polypeptide skeleton or comprises a single-chain polypeptide skeleton. In some embodiments, the GLP-1 receptor-amyrin receptor agonist may further comprise one to three extensions, which may be covalently bonded to lysine (Lys,K) residues.

[0094] In some embodiments, the C-terminal amide portion of the amyrin receptor agonist is free to allow the amyrin receptor agonist to maintain maximum biological activity. Therefore, in some embodiments, the C-terminal residue of the GLP-1 receptor agonist is covalently bonded to the N-terminal residue of either an optional peptide linker or the amyrin receptor agonist, and the N-terminal residue of the amyrin receptor agonist is covalently bonded to the C-terminus of either an optional peptide linker or the GLP-1 receptor agonist. In some embodiments, the peptide includes a linker. In some embodiments, the linker is a peptide linker. In some embodiments, the linker contains 1 to 30 native amino acids, e.g., 3 to 6 amino acids, e.g., 4 to 5 amino acids, e.g., 5 amino acids. In some embodiments, the polypeptide backbone of the GLP-1 receptor-amyrin receptor agonist comprises one, two, or three lysine residues. In some embodiments, the polypeptide backbone of the GLP-1 receptor-amyrin receptor agonist comprises one or two lysine residues. In some embodiments, the polypeptide backbone of the GLP-1 receptor-amyrin receptor agonist comprises one lysine residue.

[0095] In some embodiments, the GLP-1 receptor-amyrin receptor agonist comprises a GLP-1 receptor agonist linked to an amyrin receptor agonist. In some embodiments, the linker is one or more glycine residues, or contains them. In some embodiments, the linker contains 1 to 6 glycine residues (SEQ ID NO: 101). In some embodiments, the linker is four glycine residues (SEQ ID NO: 102), or contains them. In some embodiments, the linker is a GGGGE linker (SEQ ID NO: 103). In some embodiments, the linker is a γGlu-2xOEG linker.

[0096] In some embodiments, the polypeptide skeleton of the GLP-1 receptor-amyrin receptor agonist provided in this disclosure may be referred to as "R1" and may be described by the following formula III:Z1-Z2-Z3 (wherein Z1 is a GLP-1 receptor agonist peptide, Z2 is an optional peptide linker, and Z3 is an amyrin receptor agonist peptide). In some embodiments, Z1 may contain up to nine amino acid modifications compared to wild-type GLP-1(7-37) (SEQ ID NO: 7). In some embodiments, the C-terminus of Z1 either binds to Z2 if Z2 is present, or to Z3 if Z2 is absent. In some embodiments, Z2 is a peptide linker. In some embodiments, if Z2 is present, its N-terminus binds to the C-terminus of Z1, and its C-terminus binds to the N-terminus of Z3. In some embodiments, Z1 is or contains H-Aib-EGTFTSDVSSYLEEQAAREFIAWLVRGRK (SEQ ID NO: 91). In some embodiments, the amyrin receptor agonist comprises the peptide "Z3". In some embodiments, the amyrin receptor agonist comprises a C-terminal amide. While not bound by any particular theory, the C-terminal amide may be important to the biological activity of the amyrin receptor agonist. In some embodiments, Z3 comprises up to seven amino acid modifications relative to SEQ ID NO: 92 (ASELSTAALGRLSAELHELATLPRTETGSGSP). In some embodiments, the C-terminus of Z3 is modified with an amide group. In some embodiments, the N-terminus of Z3 is bound to the C-terminus of Z2 if Z2 is present, or to the C-terminus of Z1 if Z2 is absent. In some embodiments, Z1-Z2-Z3 comprises one, two, or three lysine and / or cysteine ​​residues. Each lysine and / or cysteine ​​residue is covalently bound to an extension portion which may be referred to herein as "R2-R3", where "R2" is an optional linker and "R3" is an extension factor.

[0097] In some embodiments, the polypeptide backbone of the dual GLP-1 receptor-amyrin receptor agonist is an amino acid sequence having at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of H-Aib-EGTFTSDVSSYLEEQAAREFIAWLVRGRKGGGGEASELSTAALGRLSAELHELATLPRTETGSGSP (SEQ ID NO: 3), or includes such an amino acid sequence. In some embodiments, the GLP-1 receptor-amyrin receptor agonist contains the unnatural amino acid 2-aminoisobutyric acid in the N-terminal portion of the GLP-1 sequence. In some embodiments, the GLP-1 receptor-amyrin receptor agonist is acylated at a lysine residue in the GLP-1 receptor agonist portion. In some embodiments, the GLP-1 receptor-amyrin receptor agonist contains a C16 diacid linked via a linker. In some embodiments, the GLP-1 receptor-amyrin receptor agonist comprises a C18 diacid conjugated via a linker. In some embodiments, the GLP-1 receptor-amyrin receptor agonist comprises a C20 diacid conjugated via a linker. In some embodiments, the GLP-1 receptor-amyrin receptor agonist comprises a C22 diacid conjugated via a linker. In some embodiments, the linker (R2) is a γGlu-2xOEG linker. In some embodiments, the dual GLP-1 receptor-amyrin receptor agonist is or comprises H-Aib-EGTFTSDVSSYLEEQAAREFIAWLVRGR-K([2-[2-[[2-[2-[2-[2-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino)butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl])-GGGGEASELSTAALGRLSAELHELATLPRTETGSGSP-amide (SEQ ID NO: 104).

[0098] In some embodiments, the dual GLP-1 receptor-amyrin receptor agonist comprises or follows formula IV (SEQ ID NO: 108): [ka] or containing a pharmaceutically acceptable salt thereof.

[0099] The dual GLP-1 receptor-amyrin receptor co-agonist is H-Aib-EGTFTSDVSSYLEEQAAREFIAWLVRGR-K([2-[2-[[2-[2-[2-[2-[[(4S)-4-carboxy-4-(19-carboxynonadecanoylamino)butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl])-GGGGEASELSTAALGRLSAELHELATLPRTETGSGSP-amide (SEQ ID NO: 105) or contains the same.

[0100] In some embodiments, the dual GLP-1 receptor-amyrin receptor agonist comprises or follows formula V (SEQ ID NO: 109): [ka] or a pharmaceutically acceptable salt thereof, or containing thereof.

[0101] In some embodiments, GLP-1 receptor amyrin receptor agonists exhibit a variety of properties that make them useful as pharmaceuticals, as described herein.

[0102] In some embodiments, GLP-1 receptor-amyrin receptor agonists are potent against GLP-1 receptors and amyrin receptors, e.g., amyrin receptor 3. The in vitro potency of GLP-1 receptor-amyrin receptor agonists against GLP-1 receptors and amyrin-3 receptors can be measured as described in assays 1 (human GLP-1 receptor in vitro potency assay) and 2 (human amyrin receptor in vitro potency assay) of WO / 2022 / 129526, respectively. WO / 2022 / 129526 is incorporated herein by reference. The potency of a compound may be described by its EC50 value. EC50 represents the concentration of the compound at which 50% of its maximum effect is observed. A lower EC50 value indicates a more potent compound. In some embodiments, the GLP-1 receptor agonists disclosed herein have an EC50 value of less than 200 pM, for example, less than 150 pM, preferably less than 100 pM, for example less than 75 pM, and more preferably less than 50 pM, for example less than 40 pM, for example less than 30 pM, for example less than 20 pM, for example less than 10 pM, for example less than 6, relative to the human GLP-1 receptor 3. In some embodiments, the GLP-1 receptor agonists disclosed herein have an EC50 value of less than 200 pM, for example less than 150 pM, preferably less than 100 pM, for example less than 75 pM, and more preferably less than 50 pM, for example less than 40 pM, for example less than 30 pM, for example less than 20 pM, for example less than 10 pM, for example less than 5, relative to the human amyrin receptor 3. In some embodiments, GLP-1 receptor-amyrin receptor agonists administered to a subject in combination with an anti-ALK7 antibody or its antigen-binding fragment are useful for total body weight reduction, such as fat weight reduction, while preventing lean body mass reduction.

[0103] Anti-GIP receptor antibody or its antigen-binding fragment Glucose-dependent insulinotropic polypeptide (GIP), a prominent incretin other than GLP-1, improves glycemic control by stimulating insulin secretion in response to nutrients (fats, glucose). Furthermore, GIP appears to improve plasma lipid profiles and stimulate calcium accumulation in bone. In contrast to GLP-1, the incretin effect of GIP is significantly reduced in patients with type 2 diabetes, but recent studies suggest that GIP efficiency can be restored after treatment in these patients, improving glucose management. Nevertheless, the role of GIP in regulating systemic metabolism beyond its direct effects on the pancreatic endocrine system remains controversial, particularly because GIP is associated with GIP action that promotes increased fat mass in animal models.

[0104] This disclosure provides combinations, such as combination therapies and / or compositions, comprising an anti-ALK7 receptor antibody or its antigen-binding fragment and an anti-GIP receptor antibody or its antigen-binding fragment. This disclosure also provides an anti-ALK7 receptor antibody or its antigen-binding fragment used in combination with an anti-GIP receptor antibody or its antigen-binding fragment for the prevention and / or treatment of overweight, obesity and / or diabetes.

[0105] In some embodiments, the second bioactive agent is an anti-GIP receptor antibody or an antigen-binding fragment thereof, or contains the same. In some embodiments, the anti-GIP receptor antibody or the antigen-binding fragment binds to the GIP receptor. In some embodiments, the anti-GIP receptor antibody or the antigen-binding fragment is characterized in that it prevents the binding of GIP to the GIP receptor. In other words, the anti-GIP receptor antibody or the antigen-binding fragment is characterized by binding to the GIP receptor without activating the GIP receptor, or without reducing its activation compared to a natural ligand such as GIP.

[0106] In some embodiments, the anti-GIP receptor antibody or its antigen-binding fragment is an antagonistic anti-GIP receptor antibody.

[0107]

[0108] In some embodiments, the anti-GIP receptor antibody or its antigen-binding fragment is a heavy chain or comprises a heavy chain. In some embodiments, the anti-GIP receptor antibody or its antigen-binding fragment heavy chain is QSLEESGGDLVRPEASLTLTCTASGFSFYTNYFLCWVRQAPGKGLEWIACIYIGAGGDTYYASWAKGRFTISKTSSTTVTLQMTSLTDADTATYFCTRYVNILDYWFHDLWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEV An amino acid sequence having at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of QFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK (Sequence ID 93), or containing such an amino acid sequence.

[0109] In some embodiments, the anti-GIP receptor antibody or its antigen-binding fragment is a light chain or comprises a light chain. In some embodiments, the anti-GIP receptor antibody or its antigen-binding fragment light chain is an amino acid sequence having at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of DVVMTQTPASVSESVGGTVTIKCQASQTIGSNLAWYQQKPGQRPKLLIYGASYLASGVPSRFKGSGSGTEYTLTISDLECADAATYYCQCTYTNNNYVGFGGGTEVVVKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 94) or comprises a light chain.

[0110] The in vitro potency of an anti-GIP receptor antibody or its antigen-binding fragment on a GIP receptor may be measured as described in Example 4. The potency of an anti-GIP receptor antibody or its antigen-binding fragment may also be described by its IC50 value, where IC50 is the concentration of the inhibitor (e.g., anti-GIP receptor antibody or its antigen-binding fragment) at which the response is halved. The lower the IC50 value, the more potent the compound. In some embodiments, the anti-GIP receptor antibodies or their antigen-binding fragments described herein have an IC50 value of less than 300 nM, e.g., less than 250 nM, e.g., less than 200 nM, e.g., less than 150 nM, e.g., less than 112 nM, relative to the human GIP receptor.

[0111] Anti-activin (Act) receptor type II antibody or its antigen-binding fragment Activin type II receptors (ActIIRs), such as ActRIIA and ActRIIB, play a role in muscle growth and fat regulation. Myostatin and activin A are proteins that negatively regulate muscle growth by promoting proteolysis, which leads to muscle atrophy. Therefore, blocking myostatin and activin A signaling promotes protein synthesis, resulting in muscle growth and reduced fat mass. Increased muscle mass is highly beneficial for improving overall metabolic health and overall well-being. Skeletal muscle is essential for exercise and is an energy-intensive tissue in the body because increasing muscle mass can prevent fat gain. Furthermore, skeletal muscle is a major site of insulin-stimulated glucose waste and is consequently negatively affected by type 2 diabetes. Therefore, identifying mechanisms to maintain / increase skeletal muscle to improve overall health is essential.

[0112] This disclosure provides combinations, such as combination therapies and / or compositions, comprising an anti-ALK7 receptor antibody or its antigen-binding fragment and an anti-ActRII antibody or its antigen-binding fragment. This disclosure also provides an anti-ALK7 receptor antibody or its antigen-binding fragment used in combination with an anti-ActRII antibody or its antigen-binding fragment for the prevention and / or treatment of overweight, obesity and / or diabetes.

[0113] In some embodiments, the second bioactive agent is an anti-ActRII antibody or its antigen-binding fragment, or contains the same. In some embodiments, the anti-ActRII antibody or its antigen-binding fragment binds to ActR, e.g., ActRII. In some embodiments, the anti-ActRII antibody or its antigen-binding fragment is characterized in that it prevents myostatin and / or activin A from binding to ActRII. In other words, the anti-ActRII antibody or its antigen-binding fragment binds to ActRII without activating it or reducing its activation compared to natural ligands such as myostatin and / or activin A. In some embodiments, the anti-ActRII antibody or its antigen-binding fragment is an antagonistic anti-ActRII antibody.

[0114]

[0115] In some embodiments, the anti-ActRII receptor antibody or its antigen-binding fragment is a heavy chain or comprises it. In some embodiments, the anti-ActRII antibody or its antigen-binding fragment heavy chain is QVQLVQSGAEVKKPGASVKVSCKASGYTFTSSYINWVRQAPGQGLEWMGTINPVSGSTSYAQKFQGRVTMTRDTSISTAYMELSRLRSDDTAVYYCARGGWFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVK An amino acid sequence having at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of FNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK (SEQ ID NO: 95), or containing the same.

[0116] In some embodiments, the anti-ActRII antibody or its antigen-binding fragment is or comprises a light chain. In some embodiments, the light chain of the anti-ActRII antibody or its antigen-binding fragment is or comprises an amino acid sequence having at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of QSALTQPASVSGSPGQSITISCTGTSSDVGSYNYVNWYQQHPGKAPKLMIYGVSKRPSGVSNRFSGSKSGNTASLTISGLQAEDEADYYCGTFAGGSYYGVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS (SEQ ID NO: 96).

[0117] Usage and Method This disclosure provides an anti-ALK7 receptor antibody or its antigen-binding fragment, which is used pharmaceutically in combination with a second bioactive agent such as a GLP-1 receptor agonist; a GLP-1 receptor-amyrin receptor agonist; an anti-GIP receptor antibody or its antigen-binding fragment; and / or an anti-ActRII antibody or its antigen-binding fragment.

[0118] In some embodiments, the anti-ALK7 receptor antibody or its antigen-binding fragment is administered together with a second bioactive agent. In some embodiments, the combination comprising the anti-ALK7 receptor antibody or its antigen-binding fragment and the second bioactive agent is administered as a co-formulation. In some embodiments, the anti-ALK7 receptor antibody or its antigen-binding fragment is administered simultaneously with or sequentially with the second bioactive agent. In some embodiments, the combination comprising the anti-ALK7 receptor antibody or its antigen-binding fragment and the second bioactive agent is administered as a loose combination.

[0119] In some embodiments, the disclosure provides combination therapy. In some embodiments, the combination therapy includes administering an anti-ALK7 receptor antibody or its antigen-binding fragment in combination with a second bioactive agent to a subject in need of treatment. In some embodiments, the disclosure provides an anti-ALK7 antibody or its antigen-binding fragment in combination with a second bioactive agent for use as a pharmaceutical.

[0120] In some embodiments, combinations, e.g., combination therapies, compositions, and / or pharmaceutical compositions disclosed herein are for medical use. The combination therapies described herein may be provided in various forms, e.g., loose combinations or co-formulations. Methods for treating or preventing overweight, obesity, and / or diabetes are provided herein. In some embodiments, diabetes is type 2 diabetes.

[0121] In some embodiments, the present disclosure provides (i) an anti-ALK7 receptor antibody or its antigen-binding fragment, and (ii) a second bioactive agent described herein, for use in the prevention and / or treatment of overweight, obesity, and / or diabetes.

[0122] In some embodiments, this method for treating and / or preventing overweight, obesity, and / or diabetes is: (i) Administer an effective amount of anti-ALK7 receptor antibody or its antigen-binding fragment, (ii) administering an effective amount of the second biological agent described herein.

[0123] Uses as provided in this disclosure may involve the administration of a therapeutically effective dose of an anti-ALK7 receptor antibody or its antigen-binding fragment and a second bioactive agent. The combination and / or composition is administered to the patient in an amount sufficient to substantially inhibit the ALK7 receptor, activate the GLP-1 receptor, and / or substantially act on the amyrin receptor, as described above. An amount appropriate to achieve this is defined as a therapeutically effective dose capable of providing weight loss, prevention, and / or treatment of obesity and / or diabetes. Single or multiple doses of the combination and / or composition may be administered as needed and, for example, in a dose and frequency tolerated by the patient. The specific dose used for treatment depends on the medical condition and history of the subject, as well as other factors such as age, weight, sex, route of administration, and efficiency.

[0124] In some embodiments, the compounds used in the methods of the present disclosure may be configured to be administered by any preferred route, including oral and intravenous routes. The compounds may be provided in a pharmaceutical composition comprising the compounds and one or more pharmaceutically acceptable excipients. Formulations for oral administration may be in the form of tablets or capsules containing powder or liquid. The dose is preferably a therapeutically effective dose or (interchangeably used) an effective dose, which is sufficient to show benefit to the subject. The actual amount administered, as well as the rate of administration and its course over time, depends on the nature and severity of the disease being treated. Therapeutic prescription, e.g., determining the dosage, is the responsibility of general practitioners and other physicians, typically taking into account the disorder being treated, the individual patient's condition, the site of delivery, the method of administration, and other factors known to the physician.

[0125] In some embodiments, administration of a combination and / or composition comprising (i) an anti-ALK7 receptor antibody or its antigen-binding fragment, and (ii) a second bioactive agent provides an increase in weight loss in subjects comparable to the weight loss in subjects administered with the anti-ALK7 receptor antibody or its antigen-binding fragment or the second bioactive agent alone.

[0126] In some embodiments, administration of a combination and / or composition comprising (i) an anti-ALK7 receptor antibody or its antigen-binding fragment, and (ii) a second bioactive agent results in increased fat mass reduction while maintaining lean body mass (preventing loss of lean body mass) in subjects comparable to fat mass reduction in subjects administered with the anti-ALK7 receptor antibody or its antigen-binding fragment alone, or with the second bioactive agent alone.

[0127] This specification provides combinations, such as combination therapies, and methods for the prevention and / or treatment of overweight and / or obesity. Generally, all subjects suffering from obesity include those who are also considered to be overweight. In some embodiments, those suffering from overweight do not include those suffering from obesity. In some embodiments, subjects suffering from overweight are humans, such as adult humans or pediatric humans (including infants, children, and adolescents). In some embodiments, human subjects suffering from overweight are classified as having a BMI of 25 kg / m2 or higher, such as a BMI of 27 kg / m2 or higher. In some embodiments, human subjects suffering from overweight have a BMI in the range of 25 to less than 30 or 27 to less than 30. In some embodiments, subjects suffering from obesity are humans, such as adult humans or pediatric humans (including infants, children, and adolescents). Body Mass Index (BMI) is a measure of increased risk for individuals with parameters associated with this metabolic syndrome, based on height and weight. The formula for calculating BMI is BMI = weight (kilograms) / height (meters)². A person suffering from obesity may have a BMI of 30 kg / m² or higher, and such a person may also be referred to as obese. In some embodiments, a person suffering from obesity has a BMI of 35 or higher, or a BMI in the range of 30 to less than 40. In some embodiments, obesity is severe obesity or morbid obesity. In some embodiments, severe obesity or morbid obesity is when a person has a BMI of 40 or higher.

[0128] This specification provides combinations for use in the prevention and / or treatment of overweight, optionally in the presence of at least one weight-related comorbidity, and methods for the prevention and / or treatment of overweight, optionally in the presence of at least one weight-related comorbidity. An increase in BMI increases the risk for individuals to develop any one of a wide range of diseases or comorbidities. In some embodiments, the weight-related comorbidities are selected from the group consisting of insulin resistance, hypertension, diabetes (such as type 2 diabetes), dyslipidemia, hypercholesterolemia, and obstructive sleep apnea.

[0129] This specification provides combinations for use in reducing body weight, and methods for reducing body weight. In some embodiments, the person subjected to weight reduction may have a BMI of 25 kg / m2 or higher, such as 27 kg / m2 or higher (overweight) or 30 kg / m2 or higher (obese). In some embodiments, the person subjected to weight reduction may have a BMI of 35 kg / m2 or higher or 40 kg / m2 or higher. The term “weight reduction” may encompass the treatment or prevention of obesity and / or overweight. In some embodiments, the techniques according to this disclosure (e.g., combinations, compositions, pharmaceutical compositions, methods, and / or uses) are for use in reducing body fat mass. In some embodiments, the techniques according to this disclosure (e.g., combinations, compositions, pharmaceutical compositions, methods, and / or uses) are for use in reducing body fat mass while maintaining lean body mass.

[0130] In some embodiments, the compounds described herein may be provided as pharmaceutically acceptable salts, hydrates, or solvates.

[0131] composition This disclosure provides pharmaceutical compositions comprising (i) an anti-ALK7 receptor antibody or its antigen-binding fragment and / or (ii) a second bioactive agent, as described herein. In some embodiments, the second bioactive agent is or contains the second bioactive agent. In some embodiments, the composition comprises (i) an anti-ALK7 receptor antibody or its antigen-binding fragment and (ii) a second bioactive agent. In some embodiments, the composition comprises (i) an anti-ALK7 receptor antibody or its antigen-binding fragment and (ii) a second bioactive agent comprising (a) a GLP-1 receptor agonist such as a GLP-1 receptor-amyrin receptor agonist, (b) an anti-GIP receptor antibody or its antigen-binding fragment, and / or (c) an anti-ActRII antibody or its antigen-binding fragment.

[0132] In one embodiment, the compositions described herein are for pharmaceutical use. In several embodiments, the compositions described herein are for use in the treatment and / or prevention of overweight, obesity, and / or diabetes. In several embodiments, the compositions described herein are for use in reducing body mass while maintaining lean body mass.

[0133] Pharmaceutical composition This disclosure provides a pharmaceutical composition comprising an anti-ALK7 antibody or its antigen-binding fragment described herein, a second bioactive agent described herein, or a combination thereof. In some embodiments, the pharmaceutical composition further comprises pharmaceutically acceptable excipients, which, as used herein, include any and all solvents, dispersion media, diluents, or other liquid vehicles suitable for the desired specific dosage form, dispersants or suspension aids, surfactants, isotonic agents, thickeners or emulsifiers, preservatives, solid binders, lubricants, and the like.

[0134] The pharmaceutical compositions described herein may be administered by appropriate methods known in the art. As will be understood by those skilled in the art, the route and / or mode of administration may depend on many factors, including, for example, but not limited to, the stability and / or pharmacokinetics and / or pharmacodynamics of the pharmaceutical compositions described herein.

[0135] In some embodiments, a pharmaceutical composition comprising the anti-ALK7 antibody or its antigen-binding fragment described herein, a second bioactive agent described herein, or a combination thereof may be administered to a patient in need at several sites, such as local sites, e.g., skin or mucous membrane sites; sites that bypass absorption, e.g., intra-arterial, intra-venous, intra-cardiac; and sites involved in absorption, e.g., intracutaneous, subcutaneous, intramuscular, oral, or intra-abdominal.

[0136] In some embodiments, the pharmaceutical compositions described herein are for use as pharmaceuticals. In some embodiments, the pharmaceutical compositions described herein are for use in the prevention and / or treatment of overweight, obesity and / or diabetes. In some embodiments, the pharmaceutical compositions described herein are for use in reducing body mass while maintaining lean body mass.

[0137] combination This disclosure provides combinations such as combination therapies. In some embodiments, the combination comprises (i) an anti-ALK7 receptor antibody or its antigen-binding fragment, and (ii) a second bioactive agent as described herein. In some embodiments, the second bioactive agent is or contains the second bioactive agent. In some embodiments, the combination comprises (i) an anti-ALK7 antibody or its antigen-binding fragment, and (ii) a second bioactive agent. In some embodiments, the combination comprises (i) an anti-ALK7 receptor antibody or its antigen-binding fragment, and (ii) a second bioactive agent comprising (a) a GLP-1 receptor agonist such as a GLP-1 receptor-amyrin receptor agonist, (b) an anti-GIP receptor antibody or its antigen-binding fragment, and / or (c) an anti-ActRII antibody or its antigen-binding fragment. In some embodiments, the combinations are not formulated together. In some embodiments, the combinations are formulated together. In some embodiments, the combinations are not provided in the same composition. In some embodiments, the combinations are provided in the same composition.

[0138] In some embodiments, the present disclosure provides a combination comprising (i) an anti-ALK7 receptor antibody or its antigen-binding fragment, and (ii) a second bioactive agent comprising a GLP-1 receptor agonist.

[0139] In some embodiments, the present disclosure provides a combination comprising (i) an anti-ALK7 receptor antibody or its antigen-binding fragment, and (ii) a second bioactive agent comprising a GLP-1 receptor-amyrin receptor agonist.

[0140] In some embodiments, the present disclosure provides a combination comprising (i) an anti-ALK7 receptor antibody or an antigen-binding fragment thereof, and (ii) a second bioactive agent comprising an anti-GIP receptor antibody or an antigen-binding fragment thereof.

[0141] In some embodiments, the disclosure provides a combination comprising (i) an anti-ALK7 receptor antibody or an antigen-binding fragment thereof, and (ii) a second bioactive agent comprising an anti-ActRII antibody or an antigen-binding fragment thereof.

[0142] In some embodiments, the combinations are used in combination therapy. In some embodiments, the combinations described herein are for use as pharmaceuticals. In some embodiments, the compositions described herein are for use in the treatment or prevention of diabetes and / or obesity. In some embodiments, the combinations described herein are for use in reducing body fat mass while maintaining lean body mass.

[0143] In some embodiments, the administration of two or more drugs may be simultaneous or sequential, i.e., not at the same time. In some embodiments, the administration of two or more drugs may be simultaneous, for example, in a single composition such as a co-formulation. In some embodiments, combination therapy does not necessarily require (or necessarily require at the same time) that the individual drugs be administered together in a single composition.

[0144] Embodiment 1. An anti-ALK7 receptor antibody or its antigen-binding fragment for use in the prevention and / or treatment of overweight, obesity, and / or diabetes, in combination with a second bioactive agent.

[0145] 2. The use according to Embodiment 1, wherein an anti-ALK7 receptor antibody or its antigen-binding fragment is administered together with a second bioactive agent.

[0146] 3. The use according to Embodiment 1, wherein the anti-ALK7 receptor antibody or its antigen-binding fragment is administered simultaneously with or sequentially with the second bioactive agent.

[0147] 4. In any of the uses described in the prior embodiments, the second bioactive agent is (a) GLP-1 receptor agonists, (b) an anti-GIP receptor antibody or its antigen-binding fragment, and / or (c) Use comprising an anti-Act receptor II antibody or its antigen-binding fragment.

[0148] 5. The use described in Embodiment 4, wherein the GLP-1 receptor agonist comprises an amino acid sequence having at least 90% sequence identity with the amino acid sequence of SEQ ID NO: 2.

[0149] 6. The use described in Embodiment 5, wherein the GLP-1 receptor agonist comprises the amino acid sequence of SEQ ID NO: 2.

[0150] 7. A use according to any one of Embodiments 4 to 6, wherein the GLP-1 receptor agonist comprises the following structure (Formula I (SEQ ID NO: 106)): [ka] The amino acid sequence is SEQ ID NO: 2 or a pharmaceutically acceptable salt thereof, used.

[0151] 8. A use according to any one of Embodiments 4 to 6, wherein the GLP-1 receptor agonist comprises the following structure (Formula II (SEQ ID NO: 107)): [ka] The amino acid sequence is SEQ ID NO: 2 or a pharmaceutically acceptable salt thereof, used.

[0152] 9. The use according to any one of the embodiments described above, wherein the second bioactive agent contains a GLP-1 receptor agonist and an amyrin receptor agonist.

[0153] 10. Use according to any one of the prior embodiments, wherein the second bioactive agent comprises a GLP-1 receptor-amyrin receptor agonist.

[0154] 11. The use described in Embodiment 10, wherein the GLP-1 receptor-amyrin receptor agonist has an amino acid sequence having at least 90% sequence identity with respect to the amino acid sequence of SEQ ID NO: 3.

[0155] 12. The use described in Embodiment 11, wherein the GLP-1 receptor-amyrin receptor agonist comprises the following structure (Formula V (SEQ ID NO: 109)): [ka] A method wherein the amino acid sequence is Sequence ID No. 3 or a pharmaceutically acceptable salt thereof.

[0156] 13. The use described in Embodiment 11, wherein the GLP-1 receptor-amyrin receptor agonist comprises the following structure (Formula IV (SEQ ID NO: 108)): [ka] The amino acid sequence is SEQ ID NO: 3 or a pharmaceutically acceptable salt thereof, used.

[0157] 14. Use according to any one of Embodiments 4 to 13, wherein the anti-GIP receptor antibody or its antigen-binding fragment is (i) A heavy chain containing an amino acid sequence having at least 90% sequence identity with the amino acid sequence of SEQ ID NO: 93, and / or (ii) Use comprising a light chain having an amino acid sequence having at least 90% sequence identity with SEQ ID NO: 94.

[0158] 15. Use as described in any one of the prior embodiments, wherein the anti-ALK7 receptor antibody or its antigen-binding fragment is (i) It is an antagonist, (ii) characterized by blocking the binding of activin B to the ALK7 receptor, and / or (iii) Use characterized by blocking the formation of a hexameric signaling complex of the ALK7 receptor having an ActRIIA / B type II receptor.

[0159] 16. Use according to any one of the prior embodiments, wherein the anti-ALK7 receptor antibody or its antigen-binding fragment comprises three heavy chain CDR sequences hCDR1-3 and three light chain CDR sequences lCDR1-3. a) hCDR1 is SGNYWN (SEQ ID NO: 8) or SGSYWN (SEQ ID NO: 9) which has a maximum of one amino acid substitution. b) hCDR2 is YISFDGRNNYNPSLKN (SEQ ID NO: 10), YISFDGRTNYNPSLKN (SEQ ID NO: 11), YISFDGRTNYNPSLKS (SEQ ID NO: 12), YISFDGRRNYNPSLKN (SEQ ID NO: 13), YISFTGRTNYNPSLKN (SEQ ID NO: 14), YISFSGRNNYNPSLKN (SEQ ID NO: 15), or YISFSGRTNYNPSLKN (SEQ ID NO: 16), which have a maximum of one or two amino acid substitutions. c)hCDR3 is DYYGSEGFGY (SEQ ID NO: 17) or DYYGSEGFAY (SEQ ID NO: 18) which has a maximum of one or two amino acid substitutions. d) lCDR1 is KSSQSLLNSRTRKNYLA (SEQ ID NO: 19), KSSQSLLVSRTRKNYLA (SEQ ID NO: 20), or KSSQSLLYSRTRKNYLA (SEQ ID NO: 21), having up to one or two amino acid substitutions. e) lCDR2 is WTSTRES (SEQ ID NO: 22) or WASTRES (SEQ ID NO: 23) which has a maximum of one amino acid substitution. f) CDR3 is KQSYNLPYT (SEQ ID NO: 24) or KRSYNLPYT (SEQ ID NO: 25) with a maximum of one amino acid substitution.

[0160] 17. Use as described in any one of the prior embodiments, wherein the anti-ALK7 receptor antibody or antigen-binding fragment is i. A heavy chain variable region (HV) containing an amino acid sequence from any one of sequence numbers 26-59, and / or ii. Use of a light chain variable region (VL) containing an amino acid sequence from any one of sequence numbers 60 to 84.

[0161] 18. Use as described in any one of the prior embodiments, wherein the anti-ALK7 receptor antibody or antigen-binding fragment (i) A heavy chain containing an amino acid sequence having at least 90% sequence identity with the amino acid sequence of SEQ ID NO: 99, and / or (ii) Use comprising a light chain having an amino acid sequence having at least 90% sequence identity with SEQ ID NO: 100.

[0162] 19. Use according to any one of the preceding embodiments, wherein the diabetes is type 1 diabetes or type 2 diabetes.

[0163] 20. A method for treating human subjects who are overweight and / or have diabetes and / or obesity, (i) The step of administering an effective amount of anti-ALK7 receptor antibody or its antigen-binding fragment, (ii) A method comprising the step of administering an effective amount of a second bioactive agent.

[0164] 21. The method according to Embodiment 20, wherein an anti-ALK7 receptor antibody or its antigen-binding fragment is administered together with a second bioactive agent.

[0165] 22. The method according to Embodiment 20, wherein an anti-ALK7 receptor antibody or its antigen-binding fragment is administered simultaneously with or sequentially with a second bioactive agent.

[0166] 23. A method according to any of Embodiments 20 to 22, wherein the second biological agent is (a) GLP-1 receptor agonists, (b) an anti-GIP receptor antibody or its antigen-binding fragment, and / or (c) A method comprising an anti-Act receptor II antibody or an antigen-binding fragment thereof.

[0167] 24. A method according to Embodiment 23, wherein the GLP-1 receptor agonist comprises an amino acid sequence having at least 90% sequence identity with respect to the amino acid sequence of SEQ ID NO: 2.

[0168] 25. A method according to Embodiment 24, wherein the GLP-1 receptor agonist comprises the amino acid sequence of SEQ ID NO: 2.

[0169] 26. The method according to Embodiment 25, wherein the GLP-1 receptor agonist comprises the following structure (Formula I (SEQ ID NO: 106)), [ka] A method wherein the amino acid sequence is Sequence ID No. 2 or a pharmaceutically acceptable salt thereof.

[0170] 27. The method according to Embodiment 25, wherein the GLP-1 receptor agonist comprises the following structure (Formula II (SEQ ID NO: 107)), [ka] A method wherein the amino acid sequence is Sequence ID No. 2 or a pharmaceutically acceptable salt thereof.

[0171] 28. The method according to any one of Embodiments 20 to 27, wherein the second bioactive agent comprises a GLP-1 receptor agonist and an amyrin receptor agonist.

[0172] 29. The method according to any one of Embodiments 20 to 27, wherein the second bioactive agent contains a GLP-1 receptor-amyrin receptor agonist.

[0173] 30. A method according to Embodiment 29, wherein the GLP-1 receptor-amyrin receptor agonist comprises an amino acid sequence having at least 90% sequence identity with respect to the amino acid sequence of SEQ ID NO: 3.

[0174] 31. The method according to Embodiment 30, wherein the GLP-1 receptor-amyrin receptor agonist comprises the following structure (Formula V (SEQ ID NO: 109)): [ka] A method wherein the amino acid sequence is Sequence ID No. 3 or a pharmaceutically acceptable salt thereof.

[0175] 32. The method according to Embodiment 31, wherein the GLP-1 receptor-amyrin receptor agonist comprises the following structure (Formula IV (SEQ ID NO: 108)): [ka] A method wherein the amino acid sequence is Sequence ID No. 3 or a pharmaceutically acceptable salt thereof.

[0176] 33. A method according to any one of embodiments 24 to 32, wherein the anti-GIP receptor antibody or its antigen-binding fragment is (i) A heavy chain containing an amino acid sequence having at least 90% sequence identity with the amino acid sequence of SEQ ID NO: 93, and / or (ii) A method comprising a light chain having an amino acid sequence having at least 90% sequence identity with SEQ ID NO: 94.

[0177] 34. A method according to any one of embodiments 20 to 33, wherein the anti-ALK7 receptor antibody or its antigen-binding fragment is (i) It is an antagonist, (ii) characterized by blocking the binding of activin B to the ALK7 receptor, and / or (iii) A method characterized by blocking the formation of a hexameric signaling complex of the ALK7 receptor having an ActRIIA / B type II receptor.

[0178] 35. A method according to any one of embodiments 20 to 34, wherein the anti-ALK7 receptor antibody or its antigen-binding fragment comprises three heavy chain CDR sequences hCDR1 to 3 and three light chain CDR sequences lCDR1 to 3, a) hCDR1 is SGNYWN (SEQ ID NO: 8) or SGSYWN (SEQ ID NO: 9) having a maximum of one amino acid substitution, b) hCDR2 is YISFDGRNNYNPSLKN (SEQ ID NO: 10), YISFDGRTNYNPSLKN (SEQ ID NO: 11), YISFDGRTNYNPSLKS (SEQ ID NO: 12), YISFDGRRNYNPSLKN (SEQ ID NO: 13), YISFTGRTNYNPSLKN (SEQ ID NO: 14), YISFSGRNNYNPSLKN (SEQ ID NO: 15), or YISFSGRTNYNPSLKN (SEQ ID NO: 16), which have a maximum of one or two amino acid substitutions. c) hCDR3 is DYYGSEGFGY (SEQ ID NO: 17) or DYYGSEGFAY (SEQ ID NO: 18) having up to one or two amino acid substitutions, d) lCDR1 is KSSQSLLNSRTRKNYLA (SEQ ID NO: 19), KSSQSLLVSRTRKNYLA (SEQ ID NO: 20), or KSSQSLLYSRTRKNYLA (SEQ ID NO: 21), having up to one or two amino acid substitutions. e) lCDR2 is WTSTRES (SEQ ID NO: 22) or WASTRES (SEQ ID NO: 23) which has a maximum of one amino acid substitution, f) A method in which lCDR3 is KQSYNLPYT (SEQ ID NO: 24) or KRSYNLPYT (SEQ ID NO: 25) having a maximum of one amino acid substitution.

[0179] 36. A method according to any one of embodiments 20 to 35, wherein the anti-ALK7 receptor antibody or antigen-binding fragment is i. A heavy chain variable region (VH) containing an amino acid sequence from any one of sequence numbers 26-59, and / or ii. A method comprising a light chain variable region (VL) containing an amino acid sequence from any one of sequence numbers 60 to 84.

[0180] 37. A method according to any one of embodiments 20 to 36, wherein the anti-ALK7 receptor antibody or antigen-binding fragment is (i) A heavy chain containing an amino acid sequence having at least 90% sequence identity with the amino acid sequence of SEQ ID NO: 99, and / or (ii) A method comprising a light chain having an amino acid sequence having at least 90% sequence identity with SEQ ID NO: 100.

[0181] 38. The method according to any one of embodiments 20 to 37, wherein the diabetes is type 1 diabetes or type 2 diabetes.

[0182] 39. (i) an anti-ALK7 receptor antibody or its antigen-binding fragment, (ii) A second biological agent, (a) GLP-1 receptor agonists, (b) an anti-GIP receptor antibody or its antigen-binding fragment, and / or (c) A combination comprising a second bioactive agent, which comprises an anti-Act receptor II antibody or an antigen-binding fragment thereof.

[0183] 40. A combination according to Embodiment 39, wherein the GLP-1 receptor agonist comprises an amino acid sequence having at least 90% sequence identity with respect to the amino acid sequence of SEQ ID NO: 2.

[0184] 41. A combination of embodiments 39 to 40, wherein the GLP-1 receptor agonist comprises the amino acid sequence of SEQ ID NO: 2.

[0185] 42. A combination according to any one of embodiments 39 to 41, wherein the GLP-1 receptor agonist comprises the following structure (Formula I (SEQ ID NO: 106)): [ka] A combination in which the aforementioned amino acid sequence is Sequence ID No. 2 or a pharmaceutically acceptable salt thereof.

[0186] 43. A combination according to any one of embodiments 39 to 41, wherein the GLP-1 receptor agonist comprises the following structure (Formula II (SEQ ID NO: 107)): [ka] A combination in which the aforementioned amino acid sequence is Sequence ID No. 2 or a pharmaceutically acceptable salt thereof.

[0187] 44. The combination according to any one of embodiments 39 to 43, wherein the second bioactive agent contains a GLP-1 receptor agonist and an amyrin receptor agonist.

[0188] 45. The combination according to any one of embodiments 39 to 44, wherein the second bioactive agent contains a GLP-1 receptor-amyrin receptor agonist.

[0189] 46. ​​A combination according to Embodiment 45, wherein the GLP-1 receptor-amyrin receptor agonist comprises an amino acid sequence having at least 90% sequence identity with respect to the amino acid sequence of SEQ ID NO: 3.

[0190] 47. The combination described in Embodiment 46, wherein the GLP-1 receptor-amyrin receptor agonist comprises the following structure (Formula IV (SEQ ID NO: 108)): [ka] A combination in which the aforementioned amino acid sequence is sequence number 3 or a pharmaceutically acceptable salt thereof.

[0191] 48. The combination described in Embodiment 46, wherein the -GLP-1 receptor-amyrin receptor agonist comprises the following structure (Formula V (SEQ ID NO: 109)): [ka] A combination in which the aforementioned amino acid sequence is sequence number 3 or a pharmaceutically acceptable salt thereof.

[0192] 49. A combination according to any one of embodiments 39 to 48, wherein the anti-GIP receptor antibody or its antigen-binding fragment is (i) A heavy chain containing an amino acid sequence having at least 90% sequence identity with the amino acid sequence of SEQ ID NO: 93, and / or (ii) A combination comprising a light chain having at least 90% sequence identity with the amino acid sequence of Sequence ID No. 94.

[0193] 50. A combination according to any one of embodiments 39 to 49, wherein the anti-ALK7 receptor antibody or its antigen-binding fragment is (i) It is an antagonist, (ii) characterized by blocking the binding of activin B to the ALK7 receptor, and / or (iii) A combination characterized by blocking the formation of a hexameric signaling complex of the ALK7 receptor having an ActRIIA / B type II receptor.

[0194] 51. A combination according to any one of embodiments 39 to 50, wherein the anti-ALK7 receptor antibody or its antigen-binding fragment comprises three heavy chain CDR sequences hCDR1 to 3 and three light chain CDR sequences lCDR1 to 3. a) hCDR1 is SGNYWN (SEQ ID NO: 8) or SGSYWN (SEQ ID NO: 9) having a maximum of one amino acid substitution, b) hCDR2 is YISFDGRNNYNPSLKN (SEQ ID NO: 10), YISFDGRTNYNPSLKN (SEQ ID NO: 11), YISFDGRTNYNPSLKS (SEQ ID NO: 12), YISFDGRRNYNPSLKN (SEQ ID NO: 13), YISFTGRTNYNPSLKN (SEQ ID NO: 14), YISFSGRNNYNPSLKN (SEQ ID NO: 15), or YISFSGRTNYNPSLKN (SEQ ID NO: 16), which have a maximum of one or two amino acid substitutions. c) hCDR3 is DYYGSEGFGY (SEQ ID NO: 17) or DYYGSEGFAY (SEQ ID NO: 18) having up to one or two amino acid substitutions, d) lCDR1 is KSSQSLLNSRTRKNYLA (SEQ ID NO: 19), KSSQSLLVSRTRKNYLA (SEQ ID NO: 20), or KSSQSLLYSRTRKNYLA (SEQ ID NO: 21), having up to one or two amino acid substitutions. e) lCDR2 is WTSTRES (SEQ ID NO: 22) or WASTRES (SEQ ID NO: 23) which has a maximum of one amino acid substitution, f) A combination in which lCDR3 is KQSYNLPYT (SEQ ID NO: 24) or KRSYNLPYT (SEQ ID NO: 25) with a maximum of one amino acid substitution.

[0195] 52. A combination according to any one of embodiments 39 to 51, wherein the anti-ALK7 receptor antibody or antigen-binding fragment is i. A heavy chain variable region (VH) containing an amino acid sequence from any one of sequence numbers 26-59, and / or ii. A combination containing a light chain variable region (VL) that includes an amino acid sequence from any one of sequence numbers 60-84.

[0196] 53. A combination according to any one of embodiments 39 to 52, wherein the anti-ALK7 receptor antibody or antigen-binding fragment is (i) A heavy chain containing an amino acid sequence having at least 90% sequence identity with the amino acid sequence of SEQ ID NO: 99, and / or (ii) A combination comprising a light chain having an amino acid sequence having at least 90% sequence identity with SEQ ID NO: 100.

[0197] 54. (i) an anti-ALK7 receptor antibody or its antigen-binding fragment, (ii) A combination including a GLP-1 receptor agonist.

[0198] 55. (i) an anti-ALK7 receptor antibody or its antigen-binding fragment, (ii) A combination comprising a GLP-1 receptor-amyrin receptor agonist.

[0199] 56. (i) an anti-ALK7 receptor antibody or its antigen-binding fragment, (ii) A combination comprising an anti-GIP receptor antibody or its antigen-binding fragment.

[0200] 57. (i) an anti-ALK7 receptor antibody or an antigen-binding fragment thereof, and (ii) an anti-ActRII antibody or an antigen-binding fragment thereof, a combination comprising the same.

[0201] 58. A pharmaceutical combination comprising the combination according to any one of embodiments 39 to 57, and at least one pharmaceutically acceptable excipient.

[0202] 59. The combination according to any one of embodiments 39 to 57, and the pharmaceutical combination according to embodiment 58, for use as a medicament.

[0203] 60. The combination according to any one of embodiments 39 to 57, and the pharmaceutical combination according to embodiment 58, for use in the prevention and / or treatment of obesity.

[0204] 61. The use according to any one of embodiments 1 to 19, the method according to any one of embodiments 20 to 38, the combination according to any one of embodiments 39 to 57, and the pharmaceutical combination according to embodiment 58, for use in reducing body fat mass.

[0205] 62. The use according to any one of embodiments 1 to 19, the method according to any one of embodiments 20 to 38, the combination according to any one of embodiments 39 to 57, and the pharmaceutical combination according to embodiment 58, for use in maintaining lean body mass.

[0206] 63. Use of the combination according to any one of embodiments 39 to 57, and the pharmaceutical combination according to embodiment 58, in the manufacture of a medicament.

[0207] 64. The use according to any one of embodiments 1 to 19 and 63, as well as the pharmaceutical combination according to embodiment 58, for the prevention or treatment of obesity.

[0208] 65. The use according to any one of embodiments 1 to 19 and 63, and the pharmaceutical combination according to embodiment 58, for reducing body fat mass.

[0209] 66. The use according to embodiment 65, wherein lean body mass is preserved. [Examples]

[0210] Abbreviation List Boc: t-butyloxycarbonyl CAD: Charged Aerosol Detector CLND: Chemiluminescent Nitrogen Detection DCM: Dichloromethane DIC: Diisopropylcarbodiimide DMF: Dimethylformamide DMSO: Dimethyl sulfoxide DTT: Dithiothreitol Et2O: Diethyl ether Fmoc: 9-Fluorenylmethyloxycarbonyl HPLC: High Performance Liquid Chromatography LC: Liquid Chromatography LCMS: Liquid Chromatography-Mass Spectrometry MALDI-MS: Matrix-Assisted Laser Desorption / Ionization Mass Spectrometry MeCN: Acetonitrile MQ: Milli-Q MS: Mass Spectrometry NMP: 1-Methyl-pyrrolidin-2-one ADO: 8-Amino-3,6-dioxaoctanoic acid OtBu: tert-Butoxy Oxyma Pure®: Cyano-hydroxyimino-acetic acid ethyl ester Pbf: 2,2,4,6,7-Pentamethyldihydrobenzofuran-5-sulfonyl RP: Reversed Phase RP-HPLC: Reversed Phase High Performance Liquid Chromatography RT: Room Temperature SEC: Size exclusion chromatography SPPS: Solid-phase peptide synthesis tBu: tert-butyl TFA: Trifluoroacetic acid TIPS Triisopropylsilane Trt: Triphenylmethyl(trityl) UPLC: Ultra-high-performance liquid chromatography UV: Ultraviolet light

[0211] Example 1: Method for preparing peptides This section presents exemplary methods for solid-phase peptide synthesis (SPPS), deprotection of amino acids, cleavage of peptides from resins, purification thereof, and detection and characterization of the resulting peptides (LCMS method).

[0212] Fatty acid building blocks For the synthesis of octadecane diacitate mono-tert-butyl ester, see pages 27-28 of WO2010 / 102886, which is incorporated herein by reference. The corresponding mono-tert-butyl esters of C20 diacitates can be prepared as appropriate.

[0213] General methods of peptide synthesis C-terminal peptide amides were prepared using amine-based resins (e.g., H-Rink Amide-MBHA resin, e.g., with a loading of 0.5 mmol / g). Peptides having a C-terminal carboxylic acid were prepared using hydroxyl-derived resins (e.g., Wang resin), and optionally pre-filled with a first amino acid (e.g., Fmoc-Gly-Wang resin with a loading of 0.4 mmol / g). Unless otherwise specified, the Fmoc-protected amino acid derivatives used were from companies such as ChemImpex, Combi-Blocks, Gyros Protein Technologies, and Iris. Fmoc-Ala-OH, Fmoc-Arg(Pbf)-OH, Fmoc-Asn(Trt)-OH, Fmoc-Asp(OtBu)-OH, Fmoc-Cys(Trt)-OH, Fmoc-Gln(Trt)-OH, Fmoc-Glu(OtBu)-OH, Fmoc-Gly-OH, Fmoc-His(Trt)-OH, Fmoc-Ile-OH, F Recommended standards included moc-Leu-OH, Fmoc-Lys(Boc)-OH, Boc-Lys(Fmoc)-OH, Fmoc-Met-OH, Fmoc-Phe-OH, Fmoc-Pro-OH, Fmoc-Ser(tBu)-OH, Fmoc-Thr(tBu)-OH, Fmoc-Trp(Boc)-OH, Fmoc-Tyr(tBu)-OH, Fmoc-Val-OH, and Fmoc-D-Tyr-(tBu)-OH. Unless otherwise specified, the natural L-form of the amino acid was used.

[0214] For the binding of modular albumin-binding sites using SPPS, suitable protected structural blocks were used, but are not limited to those listed below, such as Boc-Lys(Fmoc)-OH, Fmoc-8-amino-3,6-dioxaoctanoic acid (Fmoc-Ado-OH), Boc-Lys(Fmoc)-OH, Fmoc-Glu-OtBu, Fmoc-Gly-OH, hexadecanedioic acid mono-tert-butyl ester, octadecanedioic acid mono-tert-butyl ester, or eicosanedioic acid mono-tert-butyl ester. All operations described below were performed within a synthesis scale range of 0.1 to 0.2 mmol.

[0215] Unless otherwise specified, SPPS was synthesized using a Gyros Protein Technologies SymphonyX solid-phase peptide synthesizer with Fmoc-based chemical reactions, with slight modifications to the protocol provided by the manufacturer. Mixing was achieved by occasional nitrogen bubbling. Stepwise construction was carried out using the following steps: 1) pre-expanding the resin in DMF; 2) deprotecting Fmoc in two treatments (10 minutes each) using a 20% (v / v) piperidine DMF solution; 3) washing with DMF to remove piperidine; 4) adding Fmoc-amino acids (5 equivalents) and Oxyma Pure® (5 equivalents) as 0.5 M DMF solutions, followed by the addition of collidine (10 equivalents) as a 1 M DMF solution, followed by the addition of DIF (10 equivalents) as a 0.5 M DMF solution, followed by stirring for 0.5 to 4 hours; 5) draining the liquid reagents, followed by the addition of acetic anhydride (10 equivalents) as a 1 M solution. The unreacted peptide resin was cupped by adding it as a DMF solution, and then DMF was added to achieve the final anhydride acetate concentration of 0.25 M. 6) Excess reagent was removed by washing with DMF, and 7) a final wash was performed with DCM, MeOH, and Et2O upon completion of construction. Several amino acids, including but not limited to amino acids following sterically hindered amino acids such as Arg or Thr, were coupled for a long reaction time (e.g., 4 hours) to ensure the completion of the reaction. Boc-Lys(Fmoc)-OH, Fmoc-Ado-OH, Fmoc-Glu-OtBu, octadecanediol mono-tert-butyl ester, and eicosanedioic acid mono-tert-butyl ester were coupled using the above procedure, but the coupling time was extended to 4 hours by using 10 equivalents of each amino acid or carboxylic acid-containing building block and 10 equivalents of Oxyma Pure®. Reagents such as mono-tert-butyl eicosanedioate were prepared as 0.25 M solutions in DMF to reduce their solubility in DMF.

[0216] Common cutting methods The peptide was treated with TFA / TIPS / H2O / DTT (90:2.5:2.5:2.5:2.5 vol%) for 2 hours to cleave it from the polystyrene resin, after which the solution was poured into cold diethyl ether and centrifuged. The ether was decanted, and the peptide was then washed two or more times with diethyl ether.

[0217] General method for purification and quantitation of derivatives The crude peptide was dissolved in water with occasional addition of minimal amounts of MeCN and / or DMSO and / or TFA to facilitate dissolution in a minimal volume, and this solution was then filtered, for example, through a 0.2 μm filter. The crude peptide was then purified by reverse-phase preparative HPLC (e.g., Waters Prep 150 LC) on a column containing C18-silica gel. Elution was performed with an increasing gradient of MeCN in MQ water containing 0.1% TFA. Relevant fractions were analyzed by MALDI-MS and UPLC. Fractions containing the pure target peptide were pooled. The resulting solution was analyzed (UPLC, LCMS), and the peptide derivative was quantitated using a CLND-specific HPLC detector (Ultimate-3000 Thermo-Fischer HPLC connected to an Antek 8060 CLND). The product was dispensed into glass vials. The vials were capped with a Millipore glass fiber pre-filter. Lyophilization afforded the trifluoroacetate salt of the derivative as a white solid.

[0218] Exemplary peptides used in the following examples are shown in Table 1. [Table 2]

[0219] Example 2: Method for preparing an antibody For each monoclonal antibody, the heavy and light chains were synthesized in-frame using DNA encoding the human CD33 signal peptide, followed by the mature antibody chain, and inserted between the Hind III and Eco RI sites of the mammalian expression vector pTT5 (Durocher Y, Perret S, Kamen A. (2002) High-level and high-throughput recombinant protein production by transient transfection of suspension-growing human 293-EBNA1 cells. Nucleic Acids Res 30:E9). In the resulting plasmid, the antibody chain is under the transcriptional control of the cytomegalovirus promoter.

[0220] 1000 mL cultures of suspension-compatible human embryonic kidney (Expi293F) cells (ThermoFisher Scientific) in Expi293 expression medium (ThermoFisher Scientific) were grown in a 3000 mL Erlenmeyer flask to a density of approximately 3 × 10⁶ cells / mL. For each 1000 mL culture, 500 μg of antibody heavy chain coding plasmid and 500 μg of antibody light chain coding plasmid were diluted in 50 mL of Opti-MEM I reduced serum medium (ThermoFisher Scientific). ExpiFectamine 293 reagent (ThermoFisher Scientific) was diluted to 5.1% in Opti-MEM I reduced serum medium and incubated at room temperature for 5 minutes, then added to the diluted plasmid in 52.7 mL portions. The plasmid DNA complex reaction was gently mixed and incubated at room temperature for 10–20 minutes, then gently transferred to the Expi293F culture. Subsequently, the Expi293F cultures were incubated in a CO2 shaker (orbital shaking at 85 rpm, 36.5°C, and 8% CO2). One day after transfection, 5 mL of ExpiFectamine 293 Transfection Enhancer 1 (ThermoFisher Scientific) and 50 mL of ExpiFectamine 293 Transfection Enhancer 2 (ThermoFisher Scientific) were added to each culture flask. Five days after transfection, the cells were pelletized by centrifugation and discarded. The supernatant was further filtered through a 0.22 μm polyethersulfone filter using a Corning filter system (Corning Inc.) and used for the purification of monoclonal antibodies.

[0221] Monoclonal antibodies were purified by affinity chromatography on MabSelect Sure followed by size exclusion chromatography (SEC) on Superdex 200. Briefly, 0.5–2 L of sample was loaded into a 5 mL HiTrap MabSelect SuRe (Cytiva cat.no.11.0034-95) equilibrated with 20 mM HEPES and 150 mM NaCl pH 7.4 at a flow rate of 16 column volumes (CV) per hour. After washing with 8 CV of equilibrium buffer, the antibodies were eluted with 50 mM formic acid and 150 mM NaCl pH 3.5. The eluted protein (elution volume 15 mL) was directly purified without retention time on a 320 ml Superdex 200 26 / 600 column (Cytiva cat.no. 28-9893-36). SEC was performed at a flow rate of 0.5 CV / hour using 20 mM HEPES and 150 mM NaCl pH 7.4 as running buffer. Fractions containing monomeric mAbs were pooled and analyzed by SE-UPLC and LC-MS. Exemplary antibodies are provided in Table 2. [Table 3]

[0222] Example 3: In vitro efficacy of anti-ALK7 receptor antibody blocking antibody Receptor efficacy using R GA assay A reporter gene assay (RGA) was established to measure the ability of an anti-ALK7 receptor blocking antibody to inhibit ALK7 signaling. Since stimulated ALK7 is known to phosphorylate Smad3, which binds to SBE, ALK7 signaling was measured using the luciferase gene under the control of the Smad-binding element (SBE). Therefore, ALK7 stimulation in cells containing SBE-luciferase results in increased luciferase expression. HEK293 cells (ATCC CRL-1573) were stably transfected with the SBE-luciferase reporter plasmid pGL4.48 [luc2P / SBE / Hygro] from Promega, and single-cell clones were isolated. These single-cell clones were stably transfected with the human ALK7-S270T expression plasmid based on pcDNA3.1(+), and a single-cell clone named clone 6 was isolated. The S270T mutation made ALK7 insensitive to the SB431542 inhibitor, which was subsequently used to evade stimulation of the endogenous ALK receptor.

[0223] To measure the inhibition of ALK7 signaling, HEK293 / hALK7-S270T / SBE luciferase (clone 6) cells were seeded at a density of 20,000 cells / well in a white, opaque 384-well plate. The following day, the culture medium was removed from the assay plate and the compound diluted in medium containing 1.36 μM SB431542 was added. After incubation at 37°C for 30 minutes, 0.5 nM human activin B (R&D Systems) diluted in medium containing 1.36 μM SB431542 was added, followed by incubation at 37°C for 4 hours. The cells were lysed and the luciferase substrate was added using the Steady-GLO kit (Promega). Luminescence was detected using an EnVision 2104 plate reader (PerkinElmer) with ultra-high sensitivity luminescence detection. DMEM supplemented with 10% fetal bovine serum and 1% penicillin / streptomycin was used for cell seeding and dilution of compounds and agonists. To determine the IC50 value, antibody concentration-response curves were fitted to a four-parameter logistic equation.

[0224] Primary human subcutaneous preadipocytes Primary human subcutaneous preadipocytes are isolated from human subcutaneous adipose tissue by enzymatic digestion and selective culture techniques and purchased frozen as unpassed preadipocytes in single-use cryotubes. After thawing, the cells are suspended in Preadipocyte Growth Medium-2 supplemented with fetal bovine serum, L-glutamine, and GA-1000 SingleQuots™, dispensed into 96-well plates at 10,000 cells / well, and incubated at 37°C, 5% CO2, and 90% humidity for 24 hours. The following day, the preadipocytes are differentiated by adding Growth Medium-2 containing the above complements, along with insulin, dexamethasone, indomethacin, and isobutyl-methylxanthine, and differentiation is continued for 10 days. After differentiation, the cells are starved for 24 hours in subcutaneous basal medium containing bovine serum albumin and assayed the following day.

[0225] p-SMAD3 assay To detect phospho-SMAD3 (phosphorylated on Ser423 / Ser425) in cell lysates, the AlphaLISA® SureFire® Ultra® p-SMAD3(Ser423 / Ser425) assay, a sandwich immunoassay, was used with Alpha Technology. The cells were removed from starvation medium, recombinant human activin B (ALK7 receptor ligand) was removed to a certain concentration, and ALK7 blocking antibody, serially diluted in subcutaneous basal medium containing tween, was added to the cells and incubated for 40 minutes. The medium was then removed, and lysis buffer was added to the cells. The plate was rapidly frozen to ensure proper cell lysis. 10 μl of cell lysates were transferred to an untreated AlphaPlate-384, 5 μl of acceptor bead mixture was added to the wells, incubated for 1 hour, followed by 5 μl of donor bead mixture, and incubated overnight at room temperature protected from light. Plates were read on an EnVision® plate reader (excitation 680 nm, emission 615 nm), and IC50 values ​​were calculated in Graph Pad Prism by nonlinear regression curve fitting. The IC50 values ​​of anti-ALK7 receptor antibodies (including SEQ ID NOs. 6, 97, and 98) are shown in Table 3 of this specification. [Table 4]

[0226] Example 4: In vitro efficacy of anti-GIP receptor blocking antibody Receptor potency using RGA assay A reporter gene assay (RGA) was used to measure the ability of anti-GIPR antibodies to inhibit GIP signaling. The assay measures mouse GIPR signaling using luciferase under the control of the cAMP response element (CRE), as GIPR is known to signal via heterotrimeric G protein G, which leads to stimulation of cAMP production by adenyl cyclase. Stimulation of mouse GIPR in cells containing CRE-luciferase results in increased luciferase expression.

[0227] BHK-21 cells (ATCC CCL-10) were stably transfected with a mouse GIPR expression plasmid and a CRE luciferase reporter plasmid. The mouse GIPR expression plasmid was based on pcDNA3.1(+). The reporter plasmid was pGL4.29[luc2P / CRE / Hygro] from Promega.

[0228] To measure the inhibition of mouse GIPR signaling, BHK21 / mGIPR / CRE luciferase cells were seeded at a density of 5000 cells / well in a white, opaque 384-well plate. The following day, the culture medium was removed from the assay plate and the antibody, diluted to the desired concentration, was added. After incubation at 37°C for 30 minutes, it was incubated at 37°C for 4 hours. The cells were lysed and the luciferase substrate was added using the Steady-GLO kit (Promega). Luminescence was detected using an EnVision 2104 plate reader (PerkinElmer) with ultra-high sensitivity luminescence detection.

[0229] To determine the IC50 values ​​of the antibodies, cells were treated with a range of antibody concentrations and single concentrations of mGIP that approximately corresponded to their EC80 (25 pM). The antibody concentration-response curves were normalized to the minimum response in the presence of a competing antibody control (0%) and the response in the absence of the antibody (100%), and fitted to a four-parameter logistic equation. The IC50 values ​​of the anti-GIP receptor antibodies are shown in Table 4 of this specification. [Table 5]

[0230] Example 5: Administration of anti-ALK7 receptor antibody A and GLP-1 receptor agonist to mice The purpose of this embodiment is to investigate the effects of subcutaneous injection of exemplary anti-ALK7 receptor antibodies on total body weight, fat mass, and lean body mass in diet-obese (DIO) mice, with and without co-administration with a GP-1 passive agent.

[0231] test A 4-week study in DIO mice (n=8) investigated the weight-reducing effects of anti-ALK7 inhibitory antibody (ALK7i Ab) and GLP-1 receptor agonist (GLP-1RA) when administered alone and in combination. Male C57Bl / 6J mice were initiated on a high-fat diet (HFD) at 10 weeks of age (45% HFD, study diet) and throughout the study, with the study starting at 53 weeks of age. Mice were housed in single feedings, and diet intake was measured daily. Male DIO mice were administered either anti-ALK7 antibody (10 mg / kg, subcutaneously, twice weekly), GLP-1 receptor agonist (1 nmol / kg, subcutaneously daily), or both (combination group). Control mice were administered both isotype control antibody (10 mg / kg in PBS, subcutaneously, twice weekly) and vehicle (subcutaneously daily). Body composition was measured via EchoMRI at the end of the study. 22–24 hours prior to sacrifice, mice were given a high-fat diet of 1.5 g at the time of the final dose (approximately 6 hours before the dark cycle) to induce semi-fasting. Blood glucose was measured from the tail immediately before the mice were sacrificed. The mice were then sacrificed 22–24 hours after the final dose. Sacrificial plasma was collected for insulin measurement via ELISA, and the liver was dissected and fat and lean body mass content was measured via EchoMRI. The study design is shown in Table 5. [Table 6]

[0232] The control group maintained their body weight throughout the study. The group receiving either an anti-ALK7 inhibitor antibody (ALK7i Ab) or a GLP-1 receptor agonist (GLP-1 RA) alone showed a comparable weight loss of approximately 9% at day 29, while the combination group receiving both ALK7i Ab and GLP-1 RA showed a synergistic weight loss of approximately 18% at day 29 (Figure 1A).

[0233] Weight loss with ALK7i Ab monotherapy alone was not associated with a decrease in food intake. Weight loss induced by GLP-1RA monotherapy was associated with a transient decrease in food intake. Although weight loss in the combination group was greater than in the monotherapy group, this was not associated with a further decrease in food intake compared to GLP-1RA monotherapy (Figure 1B). Mice in the control group increased fat mass but lost lean body mass. ALK7i Ab-induced total weight loss was due to a decrease in fat mass without a decrease in lean body mass, i.e., prevention of lean body mass loss during weight loss. GLP-1RA-induced total weight loss was due to both fat mass and lean body mass. The combination therapy of ALK7i Ab and GLP-1RA-induced total weight loss was due to a large decrease in fat mass without a decrease in lean body mass (Figures 1C and 1D).

[0234] Blood glucose levels were similar among the control group, the ALK7i Ab treatment group, and the GLP-1RA monotherapy group. Blood glucose levels tended to be lower in the combination therapy groups (Figure 2A).

[0235] ALK7i Ab monotherapy increased plasma insulin compared to the control group. Plasma insulin was reduced by GLP-1RA monotherapy compared to the control group. Combination therapy with ALK7i Ab and GLP-1RA resulted in comparable fasting insulin levels compared to the control group (Figure 2B).

[0236] Total liver volume, hepatic fat volume, and hepatic lean body mass increased with ALK7i Ab monotherapy compared to the control group. Total liver volume, hepatic fat volume, and hepatic lean body mass decreased with GLP-1RA and the combination ALK7i Ab and GLP-1RA treatment groups compared to the control group (Figure 2C).

[0237] These data suggest that combination therapy with ALK7i Ab and GLP-1RA may result in greater weight loss and greater total fat loss while preserving lean body mass and reducing hepatic fat mass.

[0238] Example 6: Administration of anti-ALK7 receptor antibody A and GLP-1 receptor-amyrin agonist to mice The objective of this embodiment is to investigate the effects of subcutaneous injection of an exemplary anti-ALK7 receptor antibody on total body weight, fat mass, and lean body mass in diet-obese (DIO) mice, with and without co-administration with a GLP-1 receptor-amyrin receptor agonist.

[0239] test A 4-week study in DIO mice (n=7-8) investigated the weight-reducing effect of human anti-ALK7 inhibitory antibody (ALK7i Ab) when administered alone and in combination with a GLP-1 receptor-amyrin receptor agonist (GLP1RA-AmyRA). Male C57Bl / 6J mice were started on a high-fat diet (HFD) at 10 weeks of age (45% HFD, study diet) and throughout the study, with the study starting at 46 weeks of age. Mice were housed individually, and diet intake was measured daily. DIO mice were administered either ALK7i Ab (10 mg / kg, subcutaneously twice weekly), GLP1RA-AmyRA (1 nmol / kg, subcutaneously daily), or a combination of both (combination group). Control mice were administered both the ALK7i Ab vehicle (PBS, subcutaneously twice weekly) and the GLP1RA-AmyRA vehicle (subcutaneously daily). Body composition was measured via EchoMRI at the end of the study. On the final study day, mice received the final dose of each compound or vehicle at the start of fasting, 4–6 hours before sacrifice. Blood glucose was measured from the tail immediately before the mice were sacrificed. The mice were then euthanized, plasma was collected for insulin measurement via ELISA, the liver was dissected, and fat and lean body mass content were measured via EchoMRI. The study design is shown in Table 6. [Table 7]

[0240] The control group maintained their weight during the study. All monotherapy groups resulted in weight loss with the following: GLP1RA-AmyRA 1 nmol / kg lost approximately 5%, ALK7i Ab lost approximately 9%, and GLP1RA-AmyRA 3 nmol / kg lost approximately 13%. Combination therapy resulted in superior weight loss, with combination therapy 1 resulting in approximately 17% weight loss and combination therapy 2 resulting in approximately 19% weight loss (Figure 3A).

[0241] Weight loss with ALK7i Ab monotherapy alone was not associated with a decrease in food intake. Weight loss induced by GLP1RA-AmyRA treatment was associated with a transient decrease in food intake at both dose levels, with 3 nmol / kg inducing a greater decrease in food intake compared to 1 nmol / kg. Despite combination 1 showing greater weight loss compared to GLP1RA-AmyRA 1 nmol / kg, there was no difference in food intake between GLP1RA-AmyRA 1 nmol / kg and combination 1. Similarly, despite combination 2 showing greater weight loss compared to GLP1RA-AmyRA 3 nmol / kg, there was no difference in food intake between GLP1RA-AmyRA 3 nmol / kg and combination 2 (Figure 3B). These data suggest that the synergistic increase in weight loss in the combination groups is due to a non-food intake-reducing mechanism induced by ALK7 inhibition combined with the known food intake-reducing effects of GLP1RA and AmyRA.

[0242] The control group of mice showed increased fat mass and only slight changes in lean body mass. ALK7i Ab monotherapy induced total weight loss, which was a result of fat mass loss without a decrease in lean body mass. The total weight loss induced by GLP1RA-AmyRA monotherapy was attributable to both fat mass and lean body mass, and GLP1RA-AmyR3 nmol / kg monotherapy induced greater losses of both fat mass and lean body mass compared to GLP1RA-AmyR1 nmol / kg monotherapy. Combination groups 1 and 2 had the greatest total weight loss resulting from a greater amount of fat mass loss and had a blunter loss of lean body mass compared to their respective GLP1RA-AmyR monotherapy dose groups. This is consistent with ALK7i preserving lean body mass during weight loss (Figures 3C and 3D).

[0243] Blood glucose levels were comparable among the control, ALK7i Ab monotherapy, and GLP-1RA-AmyRA (both 1 nmol / kg) monotherapy groups. Blood glucose levels tended to be lower in the two combination therapy groups compared to the two GLP-1RA-AmyRA monotherapy groups (Figure 4A).

[0244] ALK7i Ab monotherapy increased plasma insulin levels compared to the control group. Plasma insulin levels were reduced by GLP1RA-AmyRA monotherapy. Combinations 1 and 2 eliminated the increase in plasma insulin levels resulting from ALK7i Ab monotherapy alone, resulting in comparable fasting insulin levels compared to the control group (Figure 4B).

[0245] Total liver volume, hepatic fat volume, and hepatic lean body mass increased with ALK7i Ab monotherapy alone compared to the control group. Total liver volume, hepatic fat volume, and hepatic lean body mass decreased with GLP1RA-AmyR monotherapy and with both ALK7i Ab and GLP1RA-AmyRA treatment compared to the control group (Figure 4C).

[0246] These data suggest that combination therapy with ALK7i Ab and GLP1RA-AmyRA may result in greater weight loss and greater total fat loss while preserving lean body mass and reducing hepatic fat mass.

[0247] Example 7: Administration of anti-ALK7 receptor antibody A and anti-GIPR antibody to mice The purpose of this embodiment is to investigate the effects of subcutaneous injection of exemplary anti-ALK7 receptor antibodies on total body weight, fat mass, and lean body mass in diet-obese (DIO) mice, with and without co-administration with an anti-GPIP passive agent.

[0248] test A 4-week study in DIO mice (n=8) investigated the weight-reducing effect of human ALK7 receptor antibody inhibitor (ALK7i Ab) combined with mouse anti-GIP receptor inhibitor (GIPRi Ab). Male C57Bl / 6J mice were initiated on a high-fat diet (HFD) at 10 weeks of age (45% HFD, study diet) and throughout the study, with the study starting at 62 weeks of age. Mice were housed in single feedings, and diet intake was measured daily. Male DIO mice were administered either exemplary ALK7i Ab (10 mg / kg, subcutaneously twice weekly), exemplary GIPRi Ab (30 mg / kg, subcutaneously twice weekly), or a combination of both (combination group). Control mice were administered both a vehicle with ALK7i Ab (PBS; subcutaneously twice weekly) and a vehicle with GIPRi Ab (subcutaneously once weekly). Body composition was measured via EchoMRI at the end of the study. On the final study day, mice received the final dose of each compound or vehicle while fasting, 4–6 hours before sacrifice. Fasting blood glucose was measured from the tail immediately before euthanasia. The mice were then euthanized, plasma was collected via ELISA for insulin measurement, the liver was dissected, and hepatic fat and lean body mass content were measured via EchoMRI. The study design is shown in Table 7. [Table 8]

[0249] DIO mice treated with GIPRi Ab monotherapy experienced a weight loss of approximately 10%, mice treated with ALK7i Ab monotherapy experienced a weight loss of approximately 11%, and the combination group experienced a weight loss of approximately 17% (Figure 5A).

[0250] Both the GIPRi Ab and ALK7i Ab monotherapy groups had similar food intakes. The combination therapy group showed a slight decrease in food intake compared to the monotherapy group (Figure 5B).

[0251] The total weight loss induced by GIPRi Ab monotherapy was due to both fat mass and lean body mass, while the total weight loss induced by ALK7i Ab monotherapy was due only to fat mass loss and no loss of lean body mass. The combination therapy group had the greatest decrease in total body weight compared to the GIPRi Ab group because it prevented increased fat mass loss and loss of lean body mass (Figures 5C and 5D).

[0252] Blood glucose levels were comparable among the GIPRi Ab monotherapy, ALK7i Ab monotherapy, and combination therapy groups (Figure 6A).

[0253] ALK7i Ab monotherapy increased plasma insulin levels compared to GIPRi Ab monotherapy. Plasma insulin levels decreased in combination with ALK7i Ab monotherapy but increased compared to the GIPRi Ab group (Figure 6B).

[0254] Total liver volume, hepatic fat volume, and hepatic lean body mass increased in both ALK7i Ab monotherapy and combination therapy compared to the GIPRi Ab monotherapy group (Figure 6C). These data suggest that combination therapy with ALK7i Ab and GIPRi Ab may result in greater weight loss and greater total fat loss while preserving lean body mass.

[0255] Example 8: Administration of anti-ALK7 receptor antibody A and anti-activin receptor type II antibody to mice The objective of this embodiment was to investigate the effects of subcutaneous injection of an exemplary anti-ALK7 receptor antibody on total body weight, fat mass, and lean body mass in diet-induced obesity (DIO) mice, with and without co-administration with an anti-activin receptor type II antibody. Furthermore, the anti-activin receptor type II antibody does not distinguish between various types of activin receptor type II. Therefore, a secondary objective was to determine the contribution of specific anti-ALK7 receptor antibody inhibition (specific activin receptor subtype 1) to ubiquitous activin receptor inhibition on weight loss and body composition.

[0256] test A 4-week study in DIO mice (n=6-8) investigated the weight-reducing effect of human anti-ALK7 receptor antibody inhibitor (ALK7i Ab) when administered alone and in combination with anti-activin II receptor inhibitor (ActRIIi Ab). Male C57Bl / 6J mice were started on a high-fat diet (HFD) at 10 weeks of age (45% HFD, study diet) and were tested at 63 weeks of age throughout the study. Mice were housed individually, and diet intake was measured daily. DIO mice were administered either a vehicle (control), exemplary ALK7i Ab (10 mg / kg, subcutaneously twice weekly), exemplary ActRIIi Ab (20 mg / kg, subcutaneously once weekly), or a combination of both. Control mice were administered both a vehicle with ALK7i Ab (subcutaneously twice weekly) and a vehicle with ActRIIi Ab (subcutaneously twice weekly). Body composition was measured via EchoMRI at the end of the study. On the final study day, mice received the final dose of each compound or vehicle while fasting, 4–6 hours before sacrifice. Fasting blood glucose was measured from the tail immediately before euthanasia. The mice were then euthanized, plasma was collected via ELISA for insulin measurement, the liver was dissected, and fat and lean body mass content were measured via EchoMRI. The study design is shown in Table 8. [Table 9] No weight loss was observed in the control group or the ActRIIi group. DIO mice treated with ALK7i Ab experienced only about 10% weight loss, while the combination group experienced about 6% weight loss (Figure 7A).

[0257] The control, ALK7i Ab monotherapy, and combination therapy groups all had similar dietary intakes. However, ActRIIi Ab resulted in a transient increase in dietary intake (Figure 7B). Mice in the control group increased fat mass and showed only slight changes in lean body mass. ActRIIi Ab monotherapy resulted in a decrease in fat mass accompanied by an inverse increase in lean body mass, explaining the lack of effect on total body weight. The weight loss induced by ALK7i Ab monotherapy was due to a loss of fat mass without loss of lean body mass. The combination therapy group had a similar level of fat mass loss as the ALK7i Ab monotherapy group and an increase in lean body mass similar to that observed in the ActRIIi Ab monotherapy group (Figures 7C and 7D). Antiactivin receptor type II antibodies were suggested to be a cable for partially inhibiting the ALK7 receptor, as treatment with specific ALK7 receptor antibodies further increased fat mass loss. At the same time, the fat mass loss observed in the combination group was comparable to that of the ALK7i Ab group, suggesting that the fat mass loss component may be acting to target the ALK7 receptor, which is completely inhibited by ALK7i Ab (i.e., there is no additional fat mass in the combination group).

[0258] Blood glucose levels were similar among the control group, the ALK7i Ab monotherapy group, and the combination therapy group, but were elevated with ActRIIi monotherapy (Figure 8A).

[0259] ALK7i Ab monotherapy increased plasma insulin levels compared to all other groups. Plasma insulin levels decreased in combination therapy compared to ALK7i Ab monotherapy (Figure 8B). Total liver volume, hepatic fat volume, and hepatic lean body mass were increased by both ALK7i Ab and concomitant treatment compared to the control and ActRIIi Ab groups (Figure 8C).

[0260] These data demonstrate that combination therapy with ALK7i Ab and anti-activin receptor type II antibody improves body composition and induces fat loss while increasing lean body mass.

[0261] Example 9: Administration of anti-ALK7 receptor antibody B and anti-activin receptor type II antibody to mice The objective of this embodiment is to investigate the effects of subcutaneous injection of exemplary anti-ALK7 receptor antibodies on total body weight, fat mass, and lean body mass in diet-induced obesity (DIO) mice, with and without co-administration with a GLP-1 RA (Formula II) or GLP1RA-AmyRA (Formula V) agonist.

[0262] test A 4-week study in DIO mice (n=7-8) investigated the weight-reducing effects of human anti-ALK7 inhibitory antibody (ALK7i Ab) when used alone and in combination with either a BGLP-1 receptor agonist (GLP-1RA) or a monomolecule GLP-1 receptor-amyrin receptor agonist (GLP1RA-AmyRA). Male C57Bl / 6J mice were started on a high-fat diet (HFD) at 10 weeks of age (45% HFD, study diet) and continued throughout the study, with the study starting at 41 weeks of age. Mice were housed individually, and diet intake was measured daily. DIO mice were administered ALK7i Ab (5 mg / kg subcutaneously once weekly), GLP-1RA (1 nmol / kg subcutaneously daily), GLP1RA-AmyRA (1 nmol / kg subcutaneously daily), a combination of ALK7i Ab and GLP-1RA (combination 1), or a combination of ALK7i Ab and GLP1RA-AmyRA. Control mice were administered both a vehicle for ALK7i Ab (subcutaneously once weekly) and a vehicle for either GLP-1RA or GLP1RA-AmyRA (subcutaneously daily). Body composition was measured via EchoMRI at the end of the study. Mice received their final dose approximately 24 hours before sacrifice. On the final study day, mice were fasted 4-6 hours before sacrifice. Blood glucose was measured from the tail immediately before sacrifice. Next, the mice were euthanized, plasma was collected via ELISA for insulin measurement, the liver was dissected, and fat and lean body mass content were measured via EchoMRI. The study design is shown in Table 9. [Table 10]

[0263] Control mice gained 3% more body weight. Mice treated with either GLP-1RA or GLP1RA-AmyRA alone had approximately 5% less body weight compared to control mice. Mice treated with ALK7i Ab had approximately 8% less body weight compared to control mice, and both combination groups 1 and 2 had approximately 13% less body weight compared to control mice (Figure 9A).

[0264] The control, ALK7i Ab monotherapy, and combination therapy groups all had similar dietary intakes. All groups treated with either GLP-1RA or GLP1RA-AmyRA experienced a transient decrease in dietary intake. However, despite combination therapy 1 exhibiting greater weight loss compared to GLP-1RA alone, there was no difference in dietary intake between GLP-1RA and combination therapy 1. Similarly, despite combination therapy 2 exhibiting greater weight loss compared to GLP1RA-AmyRA, there was no difference in dietary intake between GLP1RA-AmyRA and combination therapy 2 (Figure 9B). These data suggest that the synergistic increase in weight loss in the combination therapy groups is due to a non-dietary intake mechanism induced by ALK7 inhibition combined with the known dietary intake-reducing effects of GLP1RA and AmyRA.

[0265] The control group of mice showed increased fat mass and a slight decrease in lean body mass. ALK7i Ab monotherapy induced total weight loss, which was a result of fat mass loss without a decrease in lean body mass. Both GLP-1RA or GLP1RA-AmyR monotherapy induced total weight loss, attributable to both fat mass and lean body mass. Combination therapy groups 1 and 2 had the greatest total weight loss resulting from a greater loss of fat mass and no loss of lean body mass. This is consistent with ALK7i preserving lean body mass during weight loss (Figures 9C and 9D).

[0266] Blood glucose levels were comparable among the control group, the GLP-1RA and GLP1RA-AmyRA monotherapy groups, and the combination therapy groups 1 and 2. ALK7i Ab monotherapy resulted in a slight increase in blood glucose (Figure 10A).

[0267] ALK7i Ab monotherapy increased plasma insulin levels compared to the overall group. Both combination therapy with ALK7i Ab and GLP-1RA or GLP-1RA-AmyRA improved the increase in insulin levels induced by ALK7i Ab monotherapy (Figure 10B).

[0268] Total liver volume, hepatic fat volume, and hepatic lean body mass increased with ALK7i Ab monotherapy compared to the control group. Total liver volume, hepatic fat volume, and hepatic lean body mass decreased with GLP-1RA and GLP1RA-AmyRA monotherapy. Combination therapy of ALK7i Ab with either GLP-1RA or GLP1RA-AmyRA improved the increase observed with ALK7i Ab monotherapy (Figure 2C).

[0269] These data suggest that combination therapy with ALK7i Ab and GLP-1RA or ALK7i Ab and GLP1RA-AmyRA may result in greater weight loss and greater total fat loss while preserving lean body mass.

[0270] Having illustrated and described certain features of the present invention herein, many modifications, substitutions, alterations, and equivalents will be conceivable to those skilled in the art. Naturally, the accompanying embodiments are intended to encompass all modifications and alterations that fall within the true spirit of the invention.

Claims

1. A pharmaceutical product for the prevention and / or treatment of overweight, obesity, and / or diabetes, comprising an anti-ALK7 receptor antibody or an antigen-binding fragment thereof, wherein the pharmaceutical product is used in combination with a second bioactive agent, the second bioactive agent comprising a GLP-1 receptor agonist.

2. The anti-ALK7 receptor antibody or its antigen-binding fragment, (i) together with the second bioactive agent, or (ii) administered simultaneously with or in succession to the second biological agent, The pharmaceutical product according to claim 1.

3. The pharmaceutical product according to claim 1, wherein the GLP-1 receptor agonist comprises an amino acid sequence having at least 90% sequence identity with respect to the amino acid sequence of SEQ ID NO:

2.

4. The GLP-1 receptor agonist has the following structure (Formula II): 【Chemistry 1】 (In the formula, the amino acid sequence is SEQ ID NO: 2) Or a pharmaceutically acceptable salt thereof, The following structure (Formula I): 【Chemistry 2】 (In the formula, the amino acid sequence is SEQ ID NO: 2) or a pharmaceutically acceptable salt A pharmaceutical product according to claim 1, comprising:

5. The pharmaceutical product according to claim 1, wherein the second biological agent comprises a GLP-1 receptor agonist and an amyrin receptor agonist.

6. The pharmaceutical product according to claim 1, wherein the second biological agent comprises a GLP-1 receptor-amyrin receptor agonist.

7. The pharmaceutical product according to claim 6, wherein the GLP-1 receptor-amyrin receptor agonist comprises an amino acid sequence, or a fragment thereof, having at least 90% sequence identity with respect to the amino acid sequence of SEQ ID NO:

3.

8. The GLP-1 receptor-amyrin receptor agonist has the following structure (formula V): 【Transformation 3】 (In the formula, the amino acid sequence is SEQ ID NO: 3) or a pharmaceutically acceptable salt Includes, or The following structure (Formula IV): 【Chemistry 4】 (In the formula, the amino acid sequence is SEQ ID NO: 3) Or a pharmaceutically acceptable salt The pharmaceutical product according to claim 6, including the above.

9. The anti-ALK7 receptor antibody or its antigen-binding fragment, (i) Is it an antagonist? (ii) characterized by blocking the binding of activin B to the ALK7 receptor, and / or (iii) The pharmaceutical product according to claim 1, characterized by blocking the formation of a hexameric signaling complex of an ALK7 receptor having an ActRIIA / B type II receptor.

10. The anti-ALK7 receptor antibody or its antigen-binding fragment comprises three heavy chain CDR sequences hCDR1-3 and three light chain CDR sequences lCDR1-3. a) hCDR1 is SGNYWN (SEQ ID NO: 8) or SGSYWN (SEQ ID NO: 9) having a maximum of one amino acid substitution, b) hCDR2 is YISFDGRNNYNPSLKN (SEQ ID NO: 10), YISFDGRTNYNPSLKN (SEQ ID NO: 11), YISFDGRTNYNPSLKS (SEQ ID NO: 12), YISFDGRRNYNPSLKN (SEQ ID NO: 13), YISFTGRTNYNPSLKN (SEQ ID NO: 14), YISFSGRNNYNPSLKN (SEQ ID NO: 15), or YISFSGRTNYNPSLKN (SEQ ID NO: 16), having up to one or two amino acid substitutions. c) hCDR3 is DYYGSEGFGY (SEQ ID NO: 17) or DYYGSEGFAY (SEQ ID NO: 18) having up to one or two amino acid substitutions, d) lCDR1 is KSSQSLLNSRTRKNYLA (SEQ ID NO: 19), KSSQSLLLVSRTRKNYLA (SEQ ID NO: 20), or KSSQSLLYSRTRKNYLA (SEQ ID NO: 21), having up to one or two amino acid substitutions. e) lCDR2 is WTSTRES (SEQ ID NO: 22) or WASTRES (SEQ ID NO: 23) having a maximum of one amino acid substitution, f) lCDR3 is KQSYNLPYT (SEQ ID NO: 24) or KRSSYNLPYT (SEQ ID NO: 25) having a maximum of one amino acid substitution. The pharmaceutical product according to claim 1.

11. The anti-ALK7 receptor antibody or its antigen-binding fragment comprises three heavy chain CDR sequences hCDR1-3 and three light chain CDR sequences lCDR1-3. a) hCDR1 is SGSYWN (Sequence ID 9), b) hCDR2 is YISFDGRTNYNPSLKN (Sequence ID 11), c) hCDR3 is DYYGSEGFGY (Sequence ID 18), d) lcDr1 is KSSQSLLVSRTRKNYLA (Sequence ID 20), e) lcDr2 is WTSTRES (Sequence ID 22), f) lcDr3 is KQSYNLPYT (Sequence ID 24). The pharmaceutical product according to claim 1.

12. The anti-ALK7 receptor antibody or antigen-binding fragment is i. A heavy chain variable region containing the amino acid sequence according to SEQ ID NO: 43, and / or ii. Light chain variable region containing the amino acid sequence according to SEQ ID NO: 78 A pharmaceutical product according to claim 1, comprising:

13. A pharmaceutical product for the prevention and / or treatment of overweight, obesity, and / or diabetes, comprising an anti-ALK7 receptor antibody or an antigen-binding fragment thereof, wherein the pharmaceutical product is used in combination with a second bioactive agent, the second bioactive agent comprising a GLP-1 receptor agonist, the anti-ALK7 receptor antibody or its antigen-binding fragment comprising a heavy chain comprising the amino acid sequence of SEQ ID NO: 99 and a light chain comprising the amino acid sequence of SEQ ID NO: 100, and the second bioactive agent is (a) The following structure (Formula II): 【Transformation 5】 (In the formula, the amino acid sequence is SEQ ID NO: 2) Or a GLP-1 receptor agonist containing a pharmaceutically acceptable salt thereof, or (b) The following structure (formula V): 【Transformation 6】 (In the formula, the amino acid sequence is SEQ ID NO: 3) Alternatively, a GLP-1 receptor-amyrin receptor agonist containing a pharmaceutically acceptable salt thereof, Pharmaceuticals.

14. The pharmaceutical product according to claim 1 or 13, wherein body fat mass is reduced and / or lean body mass is maintained.

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