Antibody conjugated polypeptide molecules and uses thereof

Abstract

The application relates to a polypeptide molecule and an antibody conjugated polypeptide molecule thereof, which comprises a targeting antibody or a functional fragment thereof and a GLP-1 receptor agonist. The antibody conjugated polypeptide molecule synergistically acts on multiple targets, achieves the effects of reducing fat and increasing muscle or improving blood lipids, and has the effects of improving blood glucose, body weight and liver fat.

Description

[0001] Priority information This application claims priority and benefit to patent application 202510550646.5 filed with the China National Intellectual Property Administration on April 29, 2025 and patent application 202511376523.0 filed with the China National Intellectual Property Administration on September 25, 2025, the entire contents of which are incorporated herein by reference. Technical Field

[0002] This application belongs to the field of biomedical technology, specifically, this application relates to a polypeptide molecule, an antibody-conjugated polypeptide molecule, and their uses. Background Technology

[0003] Metabolic diseases such as diabetes and obesity are characterized by complications and heterogeneity, with complex pathogenesis. Treatment regimens with single drugs or single targets have limited efficacy and cannot meet the multidimensional treatment needs of clinical practice. Therefore, the development of therapeutic drugs with synergistic effects of multiple targets has become a research hotspot in the field.

[0004] Glucagon-like peptide-1 (GLP-1) is an endogenous incretin polypeptide hormone secreted by L cells in the small intestine. In the pancreas, it can protect pancreatic β cells and stimulate them to release insulin in a glucose-dependent manner, effectively controlling postprandial blood glucose with a low risk of hypoglycemia. At the same time, GLP-1 also has the effects of suppressing appetite and delaying gastric emptying, which can effectively improve body weight. Based on these physiological functions, GLP-1 receptor agonists (such as smegglutide and telposide) have been widely used in the clinical treatment of obesity and diabetes.

[0005] Glucagon (GCG) is a polypeptide hormone secreted by the α-cells of the pancreas. It mainly responds to fasting or acute hypoglycemia by stimulating hepatic gluconeogenesis and glycogenolysis to raise blood glucose levels. At the same time, GCG can affect the body's metabolism and thermogenesis. Studies have shown that exogenous administration of GCG can reduce hepatic triglyceride (TG) synthesis and promote lipid breakdown. Its effects on promoting lipid metabolism, thermogenesis, and inhibiting appetite are all of positive significance for weight control in obese patients.

[0006] Glucose-dependent insulinotropic peptide (GIP) is a polypeptide hormone secreted by small intestinal K cells. Its physiological functions include promoting insulin secretion in a glucose-dependent manner and regulating lipid allogeneic storage. Although GIP agonists may show insensitivity in patients with type 2 diabetes, related studies have confirmed that diabetic patients can regain sensitivity to GIP after their blood glucose levels decrease, providing a theoretical basis for its application in the treatment of metabolic diseases.

[0007] GDF-8 (Growth Differentiation Factor 8, also known as Myostatin or MSTN) is a protein mainly expressed by skeletal muscle cells. It belongs to the transforming growth factor-β (TGF-β) superfamily. Its core function is to control muscle volume by inhibiting the proliferation of muscle cells, resulting in a reduction in muscle mass, and thus regulating skeletal muscle growth. It is a specific negative regulator of skeletal muscle growth, and its related regulatory mechanisms provide an important target for the treatment of muscle growth-related diseases.

[0008] Currently, while GLP-1 receptor agonists (such as semaglutide and telposide) can achieve significant weight loss by suppressing appetite, they are accompanied by significant muscle loss during the weight loss process. Clinical data shows that when semaglutide and telposide are used alone, lean body mass accounts for about 30% of the total weight loss. Furthermore, these drugs have significant drug dependence, requiring continuous use, and the weight rebound rate after discontinuation is high; the rebound rate one year after stopping traditional GLP-1 drugs can reach 40%-60%. Even more challenging is that obesity often occurs in conjunction with sarcopenia. For example, in elderly patients, the proportion of sarcopenic obesity exceeds 30%. Traditional GLP-1 drug treatment easily leads patients into a vicious cycle of "weight loss-sarcopenia-decreased metabolism-rebound," failing to meet the clinical treatment needs for synergistic regulation of muscle and fat.

[0009] To compensate for the aforementioned shortcomings of GLP-1 receptor agonists, antibody drugs such as Bimagrumab have gradually entered the research field. These drugs, by targeting and regulating muscle growth, can not only directly promote muscle synthesis and improve insulin sensitivity, but also, when used in combination with GLP-1 drugs, achieve a dual therapeutic effect of "fat loss without muscle loss," thereby improving the quality of weight loss and reducing the risk of weight rebound after discontinuation of the medication. Related clinical studies have also confirmed that the combination of MSTN (i.e., GDF-8) inhibitors and GLP-1 drugs can increase energy expenditure and reduce body fat while building and preserving muscle mass. Specifically, the combination of Apitegromab and telpolide can preserve an additional 54.9% of lean body mass, while the combination of Trevogrumab and smegglutide can control the percentage of lean body mass loss from 34.5% to approximately 17%, further validating the effectiveness of this combination regimen.

[0010] In existing technologies, WO2022271867A1 and WO2024064842A1 have disclosed technical solutions for the combined administration of GLP-1 peptides and GDF8 antibodies, respectively. However, these solutions involve separate administration of the two active ingredients, resulting in cumbersome administration procedures, difficulty in synchronizing the pharmacokinetic characteristics of the two active ingredients in vivo, and difficulty in precisely controlling the synergistic effect. This fails to fully leverage the synergistic therapeutic advantages of the two and still falls short of meeting the demand for efficient and convenient treatment of metabolic diseases (such as obesity and diabetes). Therefore, developing a therapeutic molecule that integrates GLP-1 peptides and GDF8-related active ingredients into a single molecule, enabling single-dose administration and fully leveraging their synergistic effect, has become a pressing technical problem to be solved in this field. Summary of the Invention

[0011] This application aims to at least partially address the technical problems existing in the prior art. To this end, this application provides a GLP-1 receptor agonist polypeptide molecule and an antibody-conjugated polypeptide molecule, which can be used for the treatment of type 2 diabetes, obesity, non-alcoholic liver disease / non-alcoholic steatohepatitis, and other metabolic disorders.

[0012] This application is based on the following discoveries of the inventors: In a first aspect, this application provides a GLP-1 receptor agonist polypeptide molecule having the structure shown in formula (I): X1-Aib-X3GTX6TSDYSIX 13 LDX 16 X 17 X 18 X 19 X 20 X 21 FX 23 X 24 X 25 LX 27 X 28 GGPSSGAPPPS(I); in: X1 is either Y or H; X3 is either Q or E; X6 is αMeF(2F) or F; X 13 For L, αMeL, or Aib; X 16 For E, K, or R; X 17 For K, I, or Q; X 18 For R or A; X 19 For Q or A; X 20 For Aib, R, K, or Q; X 21 It can be A or E; X 23 For I or V; X 24 For E or Q; X 25 For Y or W; X 27 For L or I; X 28 It can be A or E.

[0013] The GLP-1 receptor agonist polypeptide molecule of this application is one of a GLP-1 monoreceptor agonist, a GLP-1 / GIP bireceptor agonist, or a GLP-1 / GCG / GIP trireceptor agonist, and possesses good in vitro and in vivo activity. According to embodiments of this application, the C-terminus of the GLP-1 receptor agonist polypeptide molecule is amidated. According to embodiments of this application, the C-terminus of the GLP-1 receptor agonist polypeptide molecule is not amidated.

[0014] In a second aspect, this application provides an antibody-conjugated polypeptide molecule, as detailed below: In one aspect, this application provides an antibody-coupled polypeptide molecule comprising a targeting antibody or a functional fragment thereof, and a GLP-1 receptor agonist.

[0015] On the other hand, this application provides an antibody-coupled polypeptide molecule comprising a targeting antibody or a functional fragment thereof and a GLP-1 receptor agonist, wherein the antibody or functional fragment thereof targets one of activin type 2 receptor (ActRII), growth differentiation factor (GDF-8), leptin receptor (LepR), PCSK9 and cannabinoid receptor (CB1).

[0016] In another aspect, this application provides an antibody-conjugated polypeptide molecule comprising a Latent GDF-8 antibody and a GLP-1 receptor agonist; The GLP-1 receptor agonist is selected from the GLP-1 receptor agonist polypeptide molecule described in claim 1; The Latent GDF-8 antibody includes: complementation-determining regions HCDR1, HCDR2 and HCDR3 of the heavy chain variable region, and complementation-determining regions LCDR1, LCDR2 and LCDR3 of the light chain variable region; Wherein, HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 are selected from any one of the following: d1) The amino acid sequences of HCDR1, HCDR2 and HCDR3 are shown in SEQ ID NO:1, SEQ ID NO:2 and SEQ ID NO:3 respectively, and the amino acid sequences of LCDR1, LCDR2 and LCDR3 are shown in SEQ ID NO:4, SEQ ID NO:5 and SEQ ID NO:6 respectively. d2) The amino acid sequences of HCDR1, HCDR2 and HCDR3 are shown in SEQ ID NO:7, SEQ ID NO:8 and SEQ ID NO:9 respectively, and the amino acid sequences of LCDR1, LCDR2 and LCDR3 are shown in SEQ ID NO:10, SEQ ID NO:11 and SEQ ID NO:12 respectively.

[0017] In another aspect, this application provides an antibody-conjugated polypeptide molecule comprising a Latent GDF-8 antibody, a linker, and a GLP-1 receptor agonist; wherein the Latent GDF-8 antibody and the GLP-1 receptor agonist are any one of the following:

[0018] Wherein, -C(O)-CH2-* represents an acetyl unit, the * end of -C(O)-CH2-* is used to connect the LatentGDF-8 antibody, and -NH2 indicates that the hydroxyl group on the carboxyl group of the amino acid K connected to -C(O)-CH2-* is replaced by -NH2; The Latent GDF-8 antibody Ab1 includes complementarity-determining regions HCDR1, HCDR2, and HCDR3 of the heavy chain variable region, and complementarity-determining regions LCDR1, LCDR2, and LCDR3 of the light chain variable region. The amino acid sequences of HCDR1, HCDR2, and HCDR3 are shown in SEQ ID NO:1, SEQ ID NO:2, and SEQ ID NO:3, respectively, and the amino acid sequences of LCDR1, LCDR2, and LCDR3 are shown in SEQ ID NO:4, SEQ ID NO:5, and SEQ ID NO:6, respectively. The Latent GDF-8 antibody Ab1 includes complementarity-determining regions HCDR1, HCDR2, and HCDR3 of the heavy chain variable region, and complementarity-determining regions LCDR1, LCDR2, and LCDR3 of the light chain variable region. The amino acid sequences of HCDR1, HCDR2, and HCDR3 are shown in SEQ ID NO:7, SEQ ID NO:8, and SEQ ID NO:9, respectively, and the amino acid sequences of LCDR1, LCDR2, and LCDR3 are shown in SEQ ID NO:10, SEQ ID NO:11, and SEQ ID NO:12, respectively.

[0019] This application conjugates GLP-1 receptor agonist peptides to antibodies, enabling synergistic effects on multiple targets to achieve effects such as fat reduction and muscle gain, or improvement of blood lipids. It also improves blood sugar, weight, and liver fat. Furthermore, it can improve the stability of the peptides in vivo, meeting the requirements for monthly formulation dosing.

[0020] In a third aspect of this application, a composition is proposed. According to embodiments of this application, the composition comprises the antibody-conjugated polypeptide molecule described in the second aspect. The composition of this application can effectively prevent or treat metabolic disorder-related diseases, especially obesity, diabetes, dyslipidemia-related diseases, fatty liver disease, metabolic syndrome, and non-alcoholic fatty liver disease.

[0021] In a fourth aspect of this application, the application provides the use of the antibody-drug conjugate molecule described in the second aspect or the composition described in the third aspect in the preparation of a medicament for the treatment or prevention of metabolic disorder-related diseases.

[0022] Additional aspects and advantages of this application will be set forth in part in the description which follows, and in part will be obvious from the description, or may be learned by practice of this application. Attached Figure Description

[0023] Figure 1 The effects of each group on body weight and food intake of DIO mice in Example 6 of this application are shown, where *P<0.05 vs Vehicle; #P<0.05 vs Tirzepatide; Figure 2 The effects of each group on muscle and fat in DIO mice in Example 6 of this application are shown, where *P<0.05 vs Vehicle; #P<0.05 vs Tirzepatide; Figure 3 The effects of each group on body weight and food intake of DIO mice in Example 7 of this application are shown, where *P<0.05 vsVehicle; Figure 4The effects of each group on muscle and fat in DIO mice in Example 7 of this application are shown, where *P<0.05 vsVehicle. Detailed Implementation

[0024] The embodiments of this application are described in detail below. The embodiments described below are exemplary and are only used to explain this application, and should not be construed as limiting this application.

[0025] It should be noted that the terms "first" and "second" are used for descriptive purposes only and should not be construed as indicating or implying relative importance or implicitly specifying the number of technical features indicated. Therefore, a feature defined as "first" or "second" may explicitly or implicitly include one or more of that feature. Furthermore, in the description of this application, unless otherwise stated, "multiple" means two or more.

[0026] In this application, the terms "comprising" or "including" are open-ended expressions, meaning they include the content specified in this application but do not exclude other aspects.

[0027] In this application, the terms “optionally,” “optionally,” or “optionally” generally refer to an event or condition described subsequently that may but may not occur, and the description includes both cases in which the event or condition occurs and cases in which the event or condition does not occur.

[0028] In this application, the term "antibody" refers to any isotype of intact immunoglobulin, or a fragment thereof (which can compete with the intact antibody for specific binding to a target antigen), and includes, for example, chimeric antibodies, humanized antibodies, fully human antibodies, and bispecific antibodies. Therefore, "antibody" is a class of antigen-binding proteins. An intact antibody will generally contain at least two full-length heavy chains and two full-length light chains. Antibodies may be derived from a single source or may be "chimeric," meaning that different parts of the antibody may be derived from two different antibodies. Antigen-binding proteins, antibodies, or binding fragments can be generated in a hybridoma via recombinant DNA technology or by enzymatic or chemical cleavage of intact antibodies.

[0029] The term "light chain," as used with respect to antibodies or fragments thereof, includes full-length light chains and fragments thereof, which have a variable region sequence sufficient to confer binding specificity. Full-length light chains include a variable region domain (VL) and a constant region domain (CL). The variable region domain of a light chain is located at the amino terminus of the polypeptide. Light chains include κ chains and λ chains.

[0030] The term "heavy chain," as used with respect to antibodies or fragments thereof, includes the full-length heavy chain and its fragments, which have a variable region sequence sufficient to confer binding specificity. The full-length heavy chain includes a variable region domain (VH) and three constant region domains (CH1, CH2, and CH3). The VH domain is located at the amino terminus of the polypeptide, and the CH domains are located at the carboxyl terminus, with CH3 being closest to the carboxyl terminus of the polypeptide. The heavy chain can belong to any isotype, including IgG (including IgG1, IgG2, IgG3, and IgG4 subtypes), IgA (including IgA1 and IgA2 subtypes), IgM, and IgE.

[0031] In this document, the terms "complementarity-determining region," "CDR," or "CDRs" refer to highly variable regions of the heavy chain of an immunoglobulin, specifically regions containing one or more of the major amino acid residues responsible for binding affinity to recognized antigens or epitopes. In specific embodiments of this disclosure, CDRs refer to highly variable regions of the heavy chain of the antibody.

[0032] In this document, heavy chain complementarity-determining regions (heavy chain variable regions, CDRs) are referred to as "HCDRs," which include HCDR1, HCDR2, and HCDR3. Commonly used CDR numbering schemes in the art include: Kabat numbering, Chothia numbering, IMGT numbering, Chothia-Martin numbering, and AHoLesk numbering. CDR definition schemes include: Kabat definition, Chothia definition, IMGT definition, Contact definition, and AbM definition. As described herein, "Kabat numbering" and "Kabat definition" refer to the numbering and definition system described in Kabat et al., USDept. of Health and Human Services, "Sequence of Proteins of Immunological Interest" (1983). For "Chothia definition," see Chothia et al., J Mol Biol 196: 901-917 (1987). Given the amino acid sequence of the variable region of an antibody, those skilled in the art can routinely determine which residues contain a specific CDR.

[0033] Kabat et al. also defined a numbering system applicable to the variable region sequence of any antibody. Those skilled in the art can readily map this Kabat numbering system to any variable region sequence without relying on any experimental data outside the sequence itself. As used herein, “Kabat numbering” refers to the numbering system described by Kabat et al., USD ept. of Health and Human Services, “Sequence of Proteins of Immunological Interest” (1983).

[0034] In this application, the term "antigen-binding fragment" refers to a substance containing all or part of the CDR of an antibody, lacking at least some amino acids present in the full-length chain but still capable of specifically binding to an antigen. Such fragments are biologically active because they bind to the target antigen and can compete with other antigen-binding molecules (including intact antibodies) for binding to a given epitope. Examples of antigen-binding fragments include, but are not limited to, Fab, Fab', F(ab')2, Fv fragments, disulfide-stabilized Fv fragments (dsFv), (dsFv)2, bispecific dsFv (dsFv-dsFv'), disulfide-stabilized bifunctional antibodies (ds diabody), single-chain antibody molecules (scFv), and scFv dimers (bivalent bifunctional antibodies). The aforementioned antigen-binding fragments are capable of binding to the same antigen as the parent antibody.

[0035] Human GDF-8 is synthesized as a 375-amino acid precursor protein containing a signal sequence, an N-terminal propeptide domain, and a C-terminal domain considered the active molecule. Maturation of GDF-8 involves a series of cleavage processes: after intracellular synthesis, the GDF-8 precursor is first depleted by a signal peptidase to obtain Pro-GDF-8; next, the proprotein convertase Furin recognizes and cleaves a specific sequence in Pro-GDF-8 to form Latent GDF-8 (also known as latent GDF-8), a crucial step in GDF-8 maturation; finally, the metalloproteinase BMP-1 or Tolloid enzyme further processes Latent GDF-8, cleaving a specific position in the leader domain to release active GDF-8. Mature GDF-8 exists as a homodimer, a structural feature essential for its biological activity. In this application, the antibody or its functional fragment targets one of activin type 2 receptor (ActRII), growth differentiation factor (GDF-8), leptin receptor (LepR), PCSK9, and cannabinoid receptor (CB1). In some embodiments, the antibody or its functional fragment targets Pro-GDF-8, Latent GDF-8, or mature GDF-8; in some embodiments, the antibody targets Latent GDF-8.

[0036] The term "GLP-1 receptor agonist" can activate one, two, or all three of the GLP-1, GCG, and GIP receptors. It can activate only the GLP-1 receptor, two GLP-1 and GIP receptors, or all three GLP-1, GCG, and GIP receptors.

[0037] The term "antibody-conjugated polypeptide molecule" is a bioconjugated drug. It consists of an antibody with specific targeting ability, a biologically active polypeptide molecule, and a linker connecting the two, covalently linked by a biochemical method. In this application, the "antibody-conjugated polypeptide molecule" includes, but is not limited to, ActRII blocking antibody-conjugated GLP-1 / GIP dual-receptor agonist, ActRII blocking antibody-conjugated GLP-1 / GCG / GIP triple-receptor agonist, GDF-8 antibody-conjugated GLP-1 / GIP dual-receptor agonist, GDF-8 antibody-conjugated GLP-1 / GCG / GIP triple-receptor agonist, and Latent GDF-8 antibody-conjugated GLP-1 / GIP dual-receptor agonist. GDF-8 antibody conjugated with GLP-1 / GCG / GIP triple receptor agonist, PCSK9 antibody conjugated with GLP-1 / GIP dual receptor agonist, PCSK9 antibody conjugated with GLP-1 / GCG / GIP triple receptor agonist, cannabinoid receptor (CB1) blocking antibody conjugated with GLP-1 / GIP dual receptor agonist, cannabinoid receptor (CB1) blocking antibody conjugated with GLP-1 / GCG / GIP triple receptor agonist, leptin receptor (LepR) agonist antibody conjugated with GLP-1 / GIP dual receptor agonist, leptin receptor (LepR) agonist antibody conjugated with GLP-1 / GCG / GIP triple receptor agonist.

[0038] In this application, the terms "identity," "homology," or "similarity" are used to describe the percentage of identical amino acids or nucleotides between two amino acid sequences or nucleic acid sequences relative to a reference sequence, determined using conventional methods. The percentage refers to the degree to which the amino acids in two polypeptides are identical at equivalent positions when the two sequences are optimally aligned. The alignment of amino acid sequence identity percentages can be performed using various methods within the art, such as software well-known in the art, including BLAST, BLAST-2, ALIGN, MEGALIGN (DNASTAR), CLUSTALW, or CLUSTAL OMEGA. Those skilled in the art can determine appropriate parameters for the aligned sequences, including any algorithms required to achieve maximum alignment of the full length of the compared sequences.

[0039] As used herein, the terms “amino acid” and “residue” are used interchangeably and, when used in the context of peptides or polypeptides, refer to naturally occurring amino acids and synthetic amino acids, as well as amino acid analogs, amino acid mimics, and non-naturally occurring amino acids that are chemically similar to naturally occurring amino acids.

[0040] The term "naturally occurring amino acid" refers to amino acids encoded by the genetic code, as well as amino acids encoded by genetic codes that have been modified post-synthesized, such as hydroxyproline, γ-carboxyglutamic acid, and O-phosphoserine. Amino acid analogs are compounds that have the same basic chemical structure as naturally occurring amino acids (i.e., the α-carbon bonded to hydrogen, a carboxyl group, an amino group, and an R group), such as homoserine, ortholeucine, methionine sulfoxide, and methionine methylsulfonium. These analogs may have modified R groups (e.g., ortholeucine) or modified peptide backbones, but retain the same basic chemical structure as naturally occurring amino acids.

[0041] The term "non-naturally occurring amino acid" refers to a compound that has the same basic chemical structure as a naturally occurring amino acid, but is not incorporated into the growing polypeptide chain through a translation complex. "Non-naturally occurring amino acids" also includes, but is not limited to, amino acids obtained by modifying (e.g., post-translational modification) naturally encoded amino acids (including, but not limited to, 20 common amino acids) but which are not themselves naturally incorporated into the growing polypeptide chain through a translation complex.

[0042] In this application, amino acids are represented by conventional single-letter or three-letter codes for natural amino acids, as well as generally accepted three-letter codes for other α-amino acids. For example, Y represents tyrosine, and α-aminoisobutyric acid can be represented by either Aib or B. Unless otherwise specified, all amino acid residues in this application are preferably in the L-configuration.

[0043] Wherein, the Aib structure is .

[0044] In this application, the terms "(L)-α-Me-(2-F)-Phe" and "αMeF(2F)" both refer to α-methyl-2-fluorophenylalanine, with the following structural formula: .

[0045] In this application, the term "aMeL" refers to α-methyl-leucine, whose structural formula is [insert structural formula here]. .

[0046] In this application, the term "BrAc" refers to the structural unit Br-CH2-C(=O)-, namely the bromoacetyl group (or bromoacetyl group). It is a reactive functional group that can react with the thiol group (-SH) in the active site of proteins.

[0047] In this application, the term "acetyl unit (-C(O)-CH2-*)" is used as the terminal modification group of the linker, and the asterisk (*) end indicates the linker that forms a thioether bond with the coupling site on the antibody (such as the thiol group of cysteine).

[0048] In this application, the term "coupling site" refers to a specific amino acid residue on an antibody or its functional fragment that is used for covalently linking to a linker (and consequently to a peptide drug).

[0049] In this application, the term "cysteine" refers to a naturally occurring cysteine ​​residue whose side chain thiol group can be used for coupling.

[0050] In this application, the term "substituted cysteine" refers to a non-natural cysteine ​​residue (e.g., engineered cysteine) introduced into the antibody through genetic engineering mutation for site-directed conjugation. In a preferred embodiment, the conjugation site is located at position 272 (according to the EU index number) of the antibody's Fc region, and this site is a cysteine ​​residue introduced through mutation.

[0051] In this application, the term "agonist" refers to a substance (ligand) that activates the receptor type.

[0052] In this application, the term "composition" refers to a form in which the biological activity of the active ingredient is permitted and which does not contain any additional ingredients that would have unacceptable toxicity to the subject to which the composition is applied.

[0053] In this application, "pharmaceuticalally acceptable excipients" may include any solvent, carrier, excipient, diluent, or other liquid excipient, etc., suitable for the specific target dosage form. The use of any conventional excipients, except those incompatible with the antibody-drug conjugate molecule of this application, such as any adverse biological effects or harmful interactions with any other component of the pharmaceutically acceptable composition, is also within the scope of this application.

[0054] In this application, the terms "subject" or "patient" refer to a mammalian subject or patient. Exemplary subjects include, but are not limited to, humans, monkeys, dogs, cats, mice, rats, cattle, horses, camels, alpacas, poultry, goats, and sheep. In some embodiments, the subject is a human. In some embodiments, the subject is a person suspected of having cancer, an autoimmune disease or condition, and / or an infection.

[0055] In this application, the term "treatment" is used to refer to achieving a desired pharmacological and / or physiological effect. This effect may be preventative in terms of complete or partial prevention of a disease or its symptoms, and / or therapeutic in terms of partial or complete cure of a disease and / or adverse effects caused by the disease. As used herein, "treatment" covers diseases in mammals, particularly humans, including: (a) prevention of disease or the onset of a condition in individuals susceptible to disease but not yet diagnosed with the disease; (b) suppression of disease, such as inhibiting disease progression; or (c) alleviating disease, such as reducing disease-related symptoms. As used herein, "treatment" encompasses any administration of a drug or compound to an individual to treat, cure, alleviate, improve, reduce, or suppress the individual's disease, including but not limited to administering a drug containing a compound described herein to an individual in need.

[0056] GLP-1 receptor agonist peptide molecules In a first aspect, this application provides a GLP-1 receptor agonist polypeptide molecule having the structure shown in formula (I): X1-Aib-X3GTX6TSDYSIX 13 LDX 16 X 17 X 18 X 19 X 20 X 21 FX 23 X 24 X 25 LX 27 X 28 GGPSSGAPPPS(I); in: X1 is either Y or H; X3 is either Q or E; X6 is αMeF(2F) or F; X 13 For L, αMeL, or Aib; X 16 For E, K, or R; X 17 For K, I, or Q; X 18 For R or A; X 19 For Q or A; X 20 For Aib, R, K, or Q; X 21 It can be A or E; X 23 For I or V; X 24 For E or Q; X25 For Y or W; X 27 For L or I; X 28 It can be A or E.

[0057] According to embodiments of this application, the above-mentioned GLP-1 receptor agonist polypeptide molecule may further include at least one of the following technical features: According to embodiments of this application, the GLP-1 receptor agonist polypeptide molecule is a GLP-1 / GCG / GIP triad receptor agonist polypeptide molecule, and the structure shown in formula (I) is as shown in formula (Ia): X1-Aib-QGTX6TSDYSIX 13 LDX 16 X 17 X 18 QX 20 X 21 FIEX 25 LX 27 X 28 GGPSSGAPPPS(Ia); in: X1 is either Y or H; X6 is αMeF(2F) or F; X 13 For L or αMeL; X 16 It can be E or K; X 17 For K or I; X 18 For R or A; X 20 For Aib or R; X 21 It can be A or E; X 25 For Y or W; X 27 For L or I; X 28 It can be A or E.

[0058] According to embodiments of this application, the GLP-1 receptor agonist polypeptide molecule is a GLP-1 / GIP dual receptor agonist polypeptide molecule, and the structure shown in formula (I) is as shown in formula (Ib): Y-Aib-EGTFTSDYSIX 13 LDX 16 X 17 AX 19 X 20 X 21 FX 23 X24 WLX 27 AGGPSSGAPPPS(Ib); in: X 13 For L or Aib; X 16 For E, K, or R; X 17 For I or Q; X 19 For Q or A; X 20 For R, K, or Q; X 21 It can be A or E; X 23 For I or V; X 24 For E or Q; X 27 It can be L or I.

[0059] According to embodiments of this application, the GLP-1 receptor agonist polypeptide molecule has at least one of the following structural formulas: Y-Aib-QGT αMeF(2F)-TSDY SILLD EKRQ-Aib AFIEY LLAGG PSSGA PPPS (SEQ IDNO: 41); Y-Aib-QGT αMeF(2F)-TSDY SI-aMeL-LD EKRQ-Aib AFIEY LLAGG PSSGA PPPS (SEQ ID NO:42); Y-Aib-QGT αMeF(2F)TSDY SILLD KIRQ-Aib EFIEW LLAGG PSSGA PPPS (SEQ IDNO: 43); Y-Aib-QGT αMeF(2F)-TSDY SILLD KIAQR EFIEW LLAGG PSSGA PPPS (SEQ ID NO:44); Y-Aib-QGT FTSDY SILLD KIAQ-Aib EFIEW LIAGG PSSGA PPPS (SEQ ID NO:45); Y-Aib-QGT αMeF(2F)-TSDY SILLD KIAQ-Aib EFIEW LLAGG PSSGA PPPS (SEQ IDNO: 46); H-Aib-QGT FTSDY SILLD KIAQR EFIEW LLAGG PSSGA PPPS(SEQ ID NO:47); Y-Aib-QGT αMeF(2F)-TSDY SILLD KKAQ-Aib EFIEW LLAGG PSSGA PPPS(SEQ IDNO:48); Y-Aib-QGT FTSDY SILLD KIAQ-Aib EFIEW LLAGG PSSGA PPPS(SEQ ID NO:49); Y-Aib-QGT FTSDY SILLD KIAQR EFIEW LLAGG PSSGA PPPS(SEQ ID NO:50); Y-Aib-QGT FTSDY SILLD KIAQR EFIEW LLEGG PSSGA PPPS(SEQ ID NO:51); Y-Aib-QGT FTSDY SILLD KIAQR AFIEY LLAGG PSSGA PPPS(SEQ ID NO:52); Y-Aib-QGT FTSDY SILLD KIAQR AFIEY LLEGG PSSGA PPPS(SEQ ID NO:53); Y-Aib-QGT FTSDY SILLD KIAQR EFIEW LIAGG PSSGA PPPS(SEQ ID NO:54); Y-Aib-EGT FTSDY SILLD RIAQR EFIEW LLAGG PSSGA PPPS(SEQ ID NO:55); Y-Aib-EGT FTSDY SI-Aib-LD KIAQK AFVQW LIAGG PSSGA PPPS(SEQ ID NO:56); Y-Aib-EGT FTSDY SILLD EQAAR EFIEW LLAGG PSSGA PPPS(SEQ ID NO:57); Y-Aib-EGT FTSDY SILLD KIAQR EFIEW LLAGG PSSGA PPPS(SEQ ID NO:58); Y-Aib-EGT FTSDY SILLD KIAQQ EFIEW LLAGG PSSGA PPPS (SEQ ID NO: 59).

[0060] It should be noted that the "-" between amino acids in the polypeptides of the above compounds represents an amide bond. For example, in "H-Aib-Q", the "-" represents an amide bond.

[0061] According to embodiments of this application, the C-terminus of the GLP-1 receptor agonist polypeptide molecule is amidated. According to embodiments of this application, the C-terminus of the GLP-1 receptor agonist polypeptide molecule is not amidated.

[0062] Antibody-conjugated polypeptide molecules In a second aspect, this application provides an antibody-conjugated polypeptide molecule, as detailed below: In one aspect, this application provides an antibody-coupled polypeptide molecule comprising a targeting antibody or a functional fragment thereof, and a GLP-1 receptor agonist.

[0063] According to embodiments of this application, the above-mentioned antibody-conjugated polypeptide molecule may further include at least one of the following technical features: According to embodiments of this application, the targeting antibody or its functional fragment is conjugated to a GLP-1 receptor agonist.

[0064] According to embodiments of this application, the targeting antibody or its functional fragment is coupled to a GLP-1 receptor agonist via a linker.

[0065] According to embodiments of this application, the antibody-conjugated polypeptide molecule has the structure shown in formula (II): XZU(II); in: U represents the targeting antibody or its functional fragment; Z stands for connector; X is a GLP-1 receptor agonist.

[0066] According to embodiments of this application, the GLP-1 receptor agonist is selected from one of GLP-1 monoreceptor agonists, GLP-1 / GIP bireceptor agonists, and GLP-1 / GCG / GIP trireceptor agonists.

[0067] According to embodiments of this application, the GLP-1 receptor agonist is a GLP-1 / GIP dual receptor agonist.

[0068] According to embodiments of this application, the GLP-1 receptor agonist is a GLP-1 / GCG / GIP triad receptor agonist.

[0069] According to embodiments of this application, the GLP-1 receptor agonist is selected from at least one of the GLP-1 receptor agonist polypeptide molecules described in the first aspect.

[0070] According to embodiments of this application, the targeting antibody or its functional fragment targets one of activin type 2 receptor (ActRII), growth differentiation factor (GDF-8), leptin receptor (LepR), PCSK9, and cannabinoid receptor (CB1).

[0071] According to embodiments of this application, the targeting antibody or its functional fragment targets Pro-GDF-8, Latent GDF-8, or mature GDF-8; According to embodiments of this application, the targeting antibody or its functional fragment is used to target Latent GDF-8.

[0072] According to embodiments of this application, the targeted antibody is selected from at least one of full-length monoclonal antibodies, Fab antibodies, Fab' antibodies, F(ab')2 antibodies, Fv antibodies, single-chain antibodies, dsFv antibodies, and nanobodies.

[0073] According to embodiments of this application, the targeting antibody or its functional fragment is selected from full-length monoclonal antibodies or Fab antibodies.

[0074] According to embodiments of this application, the targeting antibody or its functional fragment includes: complementarity-determining regions HCDR1, HCDR2 and HCDR3 of the heavy chain variable region, and complementarity-determining regions LCDR1, LCDR2 and LCDR3 of the light chain variable region; The HCDR1, HCDR2 and HCDR3 have the HCDR1, HCDR2 and HCDR3 defined in the heavy chain variable region as shown in the amino acid sequence of SEQ ID NO:13 or SEQ ID NO:15; The LCDR1, LCDR2, and LCDR3 have LHCDR1, LCDR2, and LCDR3 defined in the heavy chain variable region as shown in the amino acid sequence of SEQ ID NO:14 or SEQ ID NO:16.

[0075] According to embodiments of this application, HCDR1, HCDR2, and HCDR3 have HCDR1, HCDR2, and HCDR3 defined in the heavy chain variable region as shown in the amino acid sequence of SEQ ID NO:13, and LCDR1, LCDR2, and LCDR3 have LHCDR1, LCDR2, and LCDR3 defined in the heavy chain variable region as shown in the amino acid sequence of SEQ ID NO:14; or The HCDR1, HCDR2, and HCDR3 have HCDR1, HCDR2, and HCDR3 defined in the heavy chain variable region as shown in the amino acid sequence of SEQ ID NO:15, and the LCDR1, LCDR2, and LCDR3 have LHCDR1, LCDR2, and LCDR3 defined in the heavy chain variable region as shown in the amino acid sequence of SEQ ID NO:16.

[0076] According to embodiments of this application, HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 are amino acid sequences obtained using the definitions of IMGT, Kabat, Chothia, AbM, or Contact.

[0077] According to embodiments of this application, the targeting antibody or its functional fragment includes: complementarity-determining regions HCDR1, HCDR2 and HCDR3 of the heavy chain variable region, and complementarity-determining regions LCDR1, LCDR2 and LCDR3 of the light chain variable region; The HCDR1 has an amino acid sequence as shown in SEQ ID NO:1 or 7, or an amino acid sequence that is at least 60% identical to SEQ ID NO:1 or 7, or an amino acid sequence that differs from SEQ ID NO:1 or 7 by 3, 2 or 1 amino acids. The HCDR2 has an amino acid sequence as shown in SEQ ID NO:2 or 8, or an amino acid sequence that is at least 80% identical to SEQ ID NO:2 or 8, or an amino acid sequence that differs from SEQ ID NO:2 or 8 by 3, 2 or 1 amino acids. The HCDR3 has an amino acid sequence as shown in SEQ ID NO:3 or 9, or an amino acid sequence that is at least 80% identical to SEQ ID NO:3 or 9, or an amino acid sequence that differs from SEQ ID NO:3 or 9 by 3, 2 or 1 amino acids. The LCDR1 has an amino acid sequence as shown in SEQ ID NO:4 or 10, or an amino acid sequence that is at least 60% identical to SEQ ID NO:4 or 10, or an amino acid sequence that differs from SEQ ID NO:4 or 10 by 3, 2 or 1 amino acids. The LCDR2 has an amino acid sequence as shown in SEQ ID NO:5 or 11, or an amino acid sequence that is at least 80% identical to SEQ ID NO:5 or 11, or an amino acid sequence that differs from SEQ ID NO:5 or 11 by 3, 2 or 1 amino acids. The LCDR3 has an amino acid sequence as shown in SEQ ID NO:6 or 12, or an amino acid sequence that is at least 80% identical to SEQ ID NO:6 or 12, or an amino acid sequence that differs from SEQ ID NO:6 or 12 by 3, 2 or 1 amino acids.

[0078] According to embodiments of this application, HCDR1, HCDR2, and HCDR3 are selected from any one of the following: a1) The HCDR1 has an amino acid sequence as shown in SEQ ID NO:1, or an amino acid sequence that is at least 60% identical to SEQ ID NO:1, or an amino acid sequence that differs from SEQ ID NO:1 by 3, 2 or 1 amino acids. The HCDR2 has an amino acid sequence as shown in SEQ ID NO:2, or an amino acid sequence that is at least 80% identical to SEQ ID NO:2, or an amino acid sequence that differs from SEQ ID NO:2 by 3, 2 or 1 amino acids. The HCDR3 has an amino acid sequence as shown in SEQ ID NO:3, or an amino acid sequence that is at least 80% identical to SEQ ID NO:3, or an amino acid sequence that differs from SEQ ID NO:3 by 3, 2 or 1 amino acids. a2) The HCDR1 has an amino acid sequence as shown in SEQ ID NO:7, or an amino acid sequence that is at least 60% identical to SEQ ID NO:7, or an amino acid sequence that differs from SEQ ID NO:7 by 3, 2 or 1 amino acids; The HCDR2 has an amino acid sequence as shown in SEQ ID NO:8, or an amino acid sequence that is at least 80% identical to SEQ ID NO:8, or an amino acid sequence that differs from SEQ ID NO:8 by 3, 2 or 1 amino acids. The HCDR3 has an amino acid sequence as shown in SEQ ID NO:9, or an amino acid sequence that is at least 80% identical to SEQ ID NO:9, or an amino acid sequence that differs from SEQ ID NO:9 by 3, 2 or 1 amino acids.

[0079] According to embodiments of this application, LCDR1, LCDR2, and LCDR3 are selected from any one of the following: b1) The LCDR1 has an amino acid sequence as shown in SEQ ID NO:4, or an amino acid sequence that is at least 60% identical to SEQ ID NO:4, or an amino acid sequence that differs from SEQ ID NO:4 by 3, 2 or 1 amino acids. The LCDR2 has an amino acid sequence as shown in SEQ ID NO:5, or an amino acid sequence that is at least 80% identical to SEQ ID NO:5, or an amino acid sequence that differs from SEQ ID NO:5 by 3, 2 or 1 amino acids. The LCDR3 has an amino acid sequence as shown in SEQ ID NO:6, or an amino acid sequence that is at least 80% identical to SEQ ID NO:6, or an amino acid sequence that differs from SEQ ID NO:6 by 3, 2 or 1 amino acids. b2) The LCDR1 has an amino acid sequence as shown in SEQ ID NO:10, or an amino acid sequence that is at least 60% identical to SEQ ID NO:10, or an amino acid sequence that differs from SEQ ID NO:10 by 3, 2 or 1 amino acids. The LCDR2 has an amino acid sequence as shown in SEQ ID NO:11, or an amino acid sequence that is at least 80% identical to SEQ ID NO:11, or an amino acid sequence that differs from SEQ ID NO:11 by 3, 2 or 1 amino acids. The LCDR3 has an amino acid sequence as shown in SEQ ID NO:12, or an amino acid sequence that is at least 80% identical to SEQ ID NO:12, or an amino acid sequence that differs from SEQ ID NO:12 by 3, 2 or 1 amino acids.

[0080] According to embodiments of this application, HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 are selected from any one of the following: c1) The HCDR1 has an amino acid sequence as shown in SEQ ID NO:1, or an amino acid sequence that is at least 60% identical to SEQ ID NO:1, or an amino acid sequence that differs from SEQ ID NO:1 by 3, 2 or 1 amino acids. The HCDR2 has an amino acid sequence as shown in SEQ ID NO:2, or an amino acid sequence that is at least 80% identical to SEQ ID NO:2, or an amino acid sequence that differs from SEQ ID NO:2 by 3, 2 or 1 amino acids. The HCDR3 has an amino acid sequence as shown in SEQ ID NO:3, or an amino acid sequence that is at least 80% identical to SEQ ID NO:3, or an amino acid sequence that differs from SEQ ID NO:3 by 3, 2 or 1 amino acids. The LCDR1 has an amino acid sequence as shown in SEQ ID NO:4, or an amino acid sequence that is at least 60% identical to SEQ ID NO:4, or an amino acid sequence that differs from SEQ ID NO:4 by 3, 2 or 1 amino acids. The LCDR2 has an amino acid sequence as shown in SEQ ID NO:5, or an amino acid sequence that is at least 80% identical to SEQ ID NO:5, or an amino acid sequence that differs from SEQ ID NO:5 by 3, 2 or 1 amino acids. The LCDR3 has an amino acid sequence as shown in SEQ ID NO:6, or an amino acid sequence that is at least 80% identical to SEQ ID NO:6, or an amino acid sequence that differs from SEQ ID NO:6 by 3, 2 or 1 amino acids. c2) The HCDR1 has an amino acid sequence as shown in SEQ ID NO:7, or an amino acid sequence that is at least 60% identical to SEQ ID NO:7, or an amino acid sequence that differs from SEQ ID NO:7 by 3, 2 or 1 amino acids. The HCDR2 has an amino acid sequence as shown in SEQ ID NO:8, or an amino acid sequence that is at least 80% identical to SEQ ID NO:8, or an amino acid sequence that differs from SEQ ID NO:8 by 3, 2 or 1 amino acids. The HCDR3 has an amino acid sequence as shown in SEQ ID NO:9, or an amino acid sequence that is at least 80% identical to SEQ ID NO:9, or an amino acid sequence that differs from SEQ ID NO:9 by 3, 2 or 1 amino acids. The LCDR1 has an amino acid sequence as shown in SEQ ID NO:10, or an amino acid sequence that is at least 60% identical to SEQ ID NO:10, or an amino acid sequence that differs from SEQ ID NO:10 by 3, 2 or 1 amino acids. The LCDR2 has an amino acid sequence as shown in SEQ ID NO:11, or an amino acid sequence that is at least 80% identical to SEQ ID NO:11, or an amino acid sequence that differs from SEQ ID NO:11 by 3, 2 or 1 amino acids. The LCDR3 has an amino acid sequence as shown in SEQ ID NO:12, or an amino acid sequence that is at least 80% identical to SEQ ID NO:12, or an amino acid sequence that differs from SEQ ID NO:12 by 3, 2 or 1 amino acids.

[0081] According to embodiments of this application, HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 are selected from any one of the following: d1) The amino acid sequences of HCDR1, HCDR2 and HCDR3 are shown in SEQ ID NO:1, SEQ ID NO:2 and SEQ ID NO:3 respectively, and the amino acid sequences of LCDR1, LCDR2 and LCDR3 are shown in SEQ ID NO:4, SEQ ID NO:5 and SEQ ID NO:6 respectively. d2) The amino acid sequences of HCDR1, HCDR2 and HCDR3 are shown in SEQ ID NO:7, SEQ ID NO:8 and SEQ ID NO:9 respectively, and the amino acid sequences of LCDR1, LCDR2 and LCDR3 are shown in SEQ ID NO:10, SEQ ID NO:11 and SEQ ID NO:12 respectively.

[0082] In this document, the amino acid sequences shown in SEQ ID NO:1 to SEQ ID NO:12 are HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 obtained according to the Kabat definition based on SEQ ID NO:13 to SEQ ID NO:16. HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 obtained based on the heavy chain variable region and light chain variable region of this application using other definition rules (e.g., IMGT, Chothia, AbM, or Contact) are also within the scope of protection of this application.

[0083] According to embodiments of this application, the heavy chain variable region contains a heavy chain framework region, the heavy chain framework region being derived from at least one of a mouse antibody, a human antibody, a primate antibody, or a mutant thereof; and / or, the light chain variable region contains a light chain framework region, the light chain framework region being derived from at least one of a mouse antibody, a human antibody, a primate antibody, or a mutant thereof.

[0084] According to embodiments of this application, the heavy chain frame region includes HFR1, HFR2, HFR3 and HFR4, and / or the light chain frame region includes LFR1, LFR2, LFR3 and LFR4.

[0085] According to an optional embodiment of this application, the amino acid sequences of HFR1, HFR2, HFR3, and HFR4 are as shown in SEQ ID NO:19, 20, 21, and 22, respectively; or The amino acid sequences of HFR1, HFR2, HFR3 and HFR4 are shown in SEQ ID NO:27, 28, 29 and 30, respectively.

[0086] According to an optional embodiment of this application, the amino acid sequences of LFR1, LFR2, LFR3, and LFR4 are as shown in SEQ ID NO:23, 24, 25, and 26, respectively; or The amino acid sequences of LFR1, LFR2, LFR3 and LFR4 are shown in SEQ ID NO:31, 32, 33 and 34, respectively.

[0087] According to an optional embodiment of this application, the amino acid sequences of HFR1, HFR2, HFR3 and HFR4 are as shown in SEQ ID NO:19, 20, 21 and 22, respectively, and the amino acid sequences of LFR1, LFR2, LFR3 and LFR4 are as shown in SEQ ID NO:23, 24, 25 and 26, respectively.

[0088] According to an optional embodiment of this application, the amino acid sequences of HFR1, HFR2, HFR3 and HFR4 are as shown in SEQ ID NO:27, 28, 29 and 30, respectively, and the amino acid sequences of LFR1, LFR2, LFR3 and LFR4 are as shown in SEQ ID NO:31, 32, 33 and 34, respectively.

[0089] According to embodiments of this application, the heavy chain variable region has an amino acid sequence as shown in SEQ ID NO: 13 or 15, or an amino acid sequence that is at least 80% identical to SEQ ID NO: 13 or 15, or an amino acid sequence that differs from SEQ ID NO: 13 or 15 by 3, 2 or 1 amino acids.

[0090] According to embodiments of this application, the light chain variable region has an amino acid sequence as shown in SEQ ID NO: 14 or 16, or an amino acid sequence having at least 80% identity with SEQ ID NO: 14 or 16, or an amino acid sequence differing from SEQ ID NO: 14 or 16 by 3, 2 or 1 amino acids.

[0091] According to an embodiment of this application, the heavy chain variable region has an amino acid sequence as shown in SEQ ID NO:13, and the light chain variable region has an amino acid sequence as shown in SEQ ID NO:14.

[0092] According to an embodiment of this application, the heavy chain variable region has an amino acid sequence as shown in SEQ ID NO:15, and the light chain variable region has an amino acid sequence as shown in SEQ ID NO:16.

[0093] According to embodiments of this application, the targeting antibody or its functional fragment further includes a heavy chain constant region and / or a light chain constant region.

[0094] According to embodiments of this application, the constant region is derived from at least one of camel-derived antibodies, mouse-derived antibodies, human-derived antibodies, primate-derived antibodies, or mutants thereof; According to embodiments of this application, the heavy chain constant region is selected from a portion or all of the heavy chain constant regions of one of IgG1, IgG2, IgG3, IgG4, IgA, IgM, IgE, or IgD.

[0095] According to embodiments of this application, the heavy chain constant region is selected from one or more of the CH1, CH2, CH3 and CH4 regions.

[0096] In some embodiments, the antibody of this application has an Fc domain (also referred to as an Fc region or Fc fragment), such as a human IgG1 Fc domain, a human IgG2 Fc domain, a human IgG3 Fc domain, or a human IgG4 Fc domain. In some embodiments, the Fc domain is wild-type. In some embodiments, the Fc domain of this disclosure is modified, for example, to increase serum half-life.

[0097] According to embodiments of this application, the heavy chain constant region of the targeting antibody or its functional fragment includes an Fc region and optionally a CH1 region.

[0098] According to an embodiment of this application, the connector comprises a GS linker peptide.

[0099] According to embodiments of this application, the GS linker peptide comprises at least one of the amino acid sequences shown in (G3S)2, (G4S)2, (G3S)3, (G4S)3, (G3S)4, (G4S)4, (G3S)5, (G4S)5, (G3S)6, and (G4S)6.

[0100] According to embodiments of this application, the connector further includes an amino acid K and / or an acetyl unit (-C(O)-CH2-).

[0101] In this paper, the acetyl unit is -C(O)-CH2-.

[0102] According to embodiments of this application, the GS linker peptide and the acetyl unit are respectively linked to two amino groups of amino acid K, and the * end of the acetyl unit (-C(O)-CH2-*) is used to link the targeting antibody or its functional fragment.

[0103] According to an embodiment of this application, the linker is selected from GS linker peptide-amino acid K-acetyl unit.

[0104] According to an embodiment of this application, the hydroxyl group on the carboxyl group of the amino acid K is replaced by -NH2.

[0105] According to an embodiment of this application, the linker is selected from GGGGSGGGGSGGGGSK(-C(O)-CH2-*)-NH2 (SEQ ID NO:60), the * end is linked to the targeting antibody or its functional fragment, the N end of GGGGSGGGGSGGGGS in the linker is linked to the GLP-1 receptor agonist, and -NH2 indicates that the hydroxyl group on the K carboxyl group of the amino acid is replaced by -NH2.

[0106] According to an embodiment of this application, one end of the connector is linked to the C-terminus of a GLP-1 receptor agonist or any lysine residue on the GLP-1 receptor agonist polypeptide chain, and the other end is linked to a targeting antibody or its functional fragment.

[0107] According to embodiments of this application, the targeting antibody or its functional fragment has one or more coupling sites, and the targeting antibody or its functional fragment is coupled to the GLP-1 receptor agonist through the coupling sites.

[0108] According to embodiments of this application, the targeting antibody or its functional fragment has two coupling sites, each coupling site being coupled to one of the GLP-1 receptor agonists.

[0109] According to embodiments of this application, the coupling site is selected from cysteine ​​or substituted cysteine.

[0110] According to embodiments of this application, the GLP-1 receptor agonist is coupled to the side chain of cysteine ​​or a substituted cysteine ​​at the coupling site.

[0111] According to an embodiment of this application, the coupling site is located on the Fc region.

[0112] According to an embodiment of this application, one end of the connector is connected to the C-terminus of a GLP-1 receptor agonist or any lysine residue on a GLP-1 receptor agonist, and the other end is connected to a cysteine ​​or a substituted cysteine ​​residue on the Fc region of the targeting antibody or its functional fragment.

[0113] According to an embodiment of this application, when one end of the connector is linked to any non-C-terminal lysine residue on a GLP-1 receptor agonist, the C-terminus of the GLP-1 receptor agonist is amidated.

[0114] According to an embodiment of this application, one end of the connector is connected to the C-terminal carboxyl group of a GLP-1 receptor agonist or the 16th or 17th lysine residue on the GLP-1 receptor agonist.

[0115] According to an embodiment of this application, the N-terminal glycine G of the linker is linked to the C-terminal carboxyl group of a GLP-1 receptor agonist or the 16th or 17th lysine residue on the GLP-1 receptor agonist.

[0116] According to an embodiment of this application, when the N-terminus of the connector is linked to the 16th or 17th lysine residue on a GLP-1 receptor agonist, the C-terminus of the GLP-1 receptor agonist is amidated.

[0117] According to an embodiment of this application, the coupling site is selected from the 272nd position on the Fc region.

[0118] According to an embodiment of this application, the 272nd position is a substituted cysteine.

[0119] In this paper, position 272 is obtained based on the human IgG1 Fc domain according to the EU number.

[0120] According to an embodiment of this application, the Fc region of the targeting antibody or its functional fragment contains an E272C mutation.

[0121] According to embodiments of this application, the amino acid sequence of the heavy chain constant region of the targeting antibody or its functional fragment is shown in SEQ ID NO:17.

[0122] According to embodiments of this application, the amino acid sequence of the light chain constant region of the targeting antibody or its functional fragment is shown in SEQ ID NO:18.

[0123] According to embodiments of this application, the heavy chain of the targeting antibody or its functional fragment has an amino acid sequence as shown in SEQ ID NO:35 or 37, or an amino acid sequence having at least 80% identity with it.

[0124] According to embodiments of this application, the light chain of the targeting antibody or its functional fragment has an amino acid sequence as shown in SEQ ID NO:36 or 38, or an amino acid sequence having at least 80% identity with it.

[0125] According to embodiments of this application, the heavy chain of the targeting antibody or its functional fragment has an amino acid sequence as shown in SEQ ID NO:35, or an amino acid sequence having at least 80% identity with it; the light chain of the targeting antibody or its functional fragment has an amino acid sequence as shown in SEQ ID NO:36, or an amino acid sequence having at least 80% identity with it.

[0126] According to embodiments of this application, the heavy chain of the targeting antibody or its functional fragment has an amino acid sequence as shown in SEQ ID NO:37, or an amino acid sequence having at least 80% identity with it; the light chain of the targeting antibody or its functional fragment has an amino acid sequence as shown in SEQ ID NO:38, or an amino acid sequence having at least 80% identity with it.

[0127] According to embodiments of this application, the heavy chain of the targeting antibody or its functional fragment has an amino acid sequence as shown in SEQ ID NO:35; the light chain of the targeting antibody or its functional fragment has an amino acid sequence as shown in SEQ ID NO:36.

[0128] According to embodiments of this application, the heavy chain of the targeting antibody or its functional fragment has an amino acid sequence as shown in SEQ ID NO:37, and the light chain of the targeting antibody or its functional fragment has an amino acid sequence as shown in SEQ ID NO:38.

[0129] On the other hand, this application provides an antibody-coupled polypeptide molecule comprising a targeting antibody or a functional fragment thereof and a GLP-1 receptor agonist, wherein the antibody or functional fragment thereof targets one of activin type 2 receptor (ActRII), growth differentiation factor (GDF-8), leptin receptor (LepR), PCSK9 and cannabinoid receptor (CB1).

[0130] According to embodiments of this application, the antibody or its functional fragment targets GDF-8.

[0131] According to embodiments of this application, the antibody or its functional fragment targets Pro-GDF-8, Latent GDF-8, or mature GDF-8.

[0132] According to embodiments of this application, the antibody or its functional fragment targets Latent GDF-8.

[0133] According to embodiments of this application, the GLP-1 receptor agonist is selected from the GLP-1 receptor agonist polypeptide molecules described in the first aspect; According to an embodiment of this application, the Latent GDF-8 antibody includes: complementarity-determining regions HCDR1, HCDR2 and HCDR3 of the heavy chain variable region, and complementarity-determining regions LCDR1, LCDR2 and LCDR3 of the light chain variable region; Wherein, HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 are selected from any one of the following: d1) The amino acid sequences of HCDR1, HCDR2 and HCDR3 are shown in SEQ ID NO:1, SEQ ID NO:2 and SEQ ID NO:3 respectively, and the amino acid sequences of LCDR1, LCDR2 and LCDR3 are shown in SEQ ID NO:4, SEQ ID NO:5 and SEQ ID NO:6 respectively. d2) The amino acid sequences of HCDR1, HCDR2 and HCDR3 are shown in SEQ ID NO:7, SEQ ID NO:8 and SEQ ID NO:9 respectively, and the amino acid sequences of LCDR1, LCDR2 and LCDR3 are shown in SEQ ID NO:10, SEQ ID NO:11 and SEQ ID NO:12 respectively.

[0134] According to embodiments of this application, the antibody-conjugated polypeptide molecule further includes a linker, through which the targeting antibody or its functional fragment and the GLP-1 receptor agonist are linked.

[0135] According to the embodiments of this application, the targeting antibody or its functional fragment and linker are as described above, and this part will not be repeated.

[0136] In another aspect, this application provides an antibody-conjugated polypeptide molecule comprising a Latent GDF-8 antibody and a GLP-1 receptor agonist; The GLP-1 receptor agonist is selected from the GLP-1 receptor agonist polypeptide molecule described in claim 1; The Latent GDF-8 antibody includes: complementation-determining regions HCDR1, HCDR2 and HCDR3 of the heavy chain variable region, and complementation-determining regions LCDR1, LCDR2 and LCDR3 of the light chain variable region; Wherein, HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 are selected from any one of the following: d1) The amino acid sequences of HCDR1, HCDR2 and HCDR3 are shown in SEQ ID NO:1, SEQ ID NO:2 and SEQ ID NO:3 respectively, and the amino acid sequences of LCDR1, LCDR2 and LCDR3 are shown in SEQ ID NO:4, SEQ ID NO:5 and SEQ ID NO:6 respectively. d2) The amino acid sequences of HCDR1, HCDR2 and HCDR3 are shown in SEQ ID NO:7, SEQ ID NO:8 and SEQ ID NO:9 respectively, and the amino acid sequences of LCDR1, LCDR2 and LCDR3 are shown in SEQ ID NO:10, SEQ ID NO:11 and SEQ ID NO:12 respectively.

[0137] According to embodiments of this application, the antibody-conjugated polypeptide molecule further includes a linker, through which the targeting antibody or its functional fragment and the GLP-1 receptor agonist are linked.

[0138] According to the embodiments of this application, the targeting antibody or its functional fragment and linker are as described above, and this part will not be repeated.

[0139] In another aspect, this application provides an antibody-conjugated polypeptide molecule comprising a Latent GDF-8 antibody, a linker, and a GLP-1 receptor agonist; wherein the Latent GDF-8 antibody and the GLP-1 receptor agonist are any one of the following:

[0140] Wherein, -C(O)-CH2-* represents an acetyl unit, the * end of -C(O)-CH2-* is used to connect the LatentGDF-8 antibody, and -NH2 indicates that the hydroxyl group in amino acid K connected to -C(O)-CH2-* is replaced by -NH2; The Latent GDF-8 antibody Ab1 includes complementarity-determining regions HCDR1, HCDR2, and HCDR3 of the heavy chain variable region, and complementarity-determining regions LCDR1, LCDR2, and LCDR3 of the light chain variable region. The amino acid sequences of HCDR1, HCDR2, and HCDR3 are shown in SEQ ID NO:1, SEQ ID NO:2, and SEQ ID NO:3, respectively, and the amino acid sequences of LCDR1, LCDR2, and LCDR3 are shown in SEQ ID NO:4, SEQ ID NO:5, and SEQ ID NO:6, respectively. The Latent GDF-8 antibody Ab1 includes complementarity-determining regions HCDR1, HCDR2, and HCDR3 of the heavy chain variable region, and complementarity-determining regions LCDR1, LCDR2, and LCDR3 of the light chain variable region. The amino acid sequences of HCDR1, HCDR2, and HCDR3 are shown in SEQ ID NO:7, SEQ ID NO:8, and SEQ ID NO:9, respectively, and the amino acid sequences of LCDR1, LCDR2, and LCDR3 are shown in SEQ ID NO:10, SEQ ID NO:11, and SEQ ID NO:12, respectively.

[0141] According to an embodiment of this application, the linker is coupled to the side chain of cysteine ​​at position 272 on the Fc region of the antibody.

[0142] According to the embodiments of this application, the targeting antibody or its functional fragment is as described above, and this part will not be repeated.

[0143] According to embodiments of this application, the antibody-coupled polypeptide molecule, Ab1-P7-1, has the following structural formula: , Wherein, S (see * for details) comes from the -SH of the cysteine ​​side chain at the antibody Ab1 coupling site; The amino acid sequence of polypeptide P7-1 is Y-Aib-QGT αMeF(2F)-TSDY SILLD EKRQ-Aib AFIEYLLAGG PSSGA PPPS. According to an embodiment of this application, the C-terminus of P7-1 is linked to a glycine G in a GS linker. According to an embodiment of this application, the linker is coupled to the side chain of cysteine ​​residue at position 272 on the Fc region of the antibody.

[0144] According to embodiments of this application, the antibody-coupled polypeptide molecule, Ab1-P7-2, has the following structural formula: , Wherein, S (see * for details) comes from the -SH of the cysteine ​​side chain at the antibody Ab1 coupling site; The amino acid sequence of polypeptide P7-2 is Y-Aib-QGT αMeF(2F)-TSDY SI-aMeL-LD EKRQ-AibAFIEY LLAGG PSSGA PPPS. According to an embodiment of this application, the C-terminus of P7-2 is linked to a glycine G in a GS linker. According to an embodiment of this application, the linker is coupled to the side chain of cysteine ​​residue at position 272 on the Fc region of the antibody.

[0145] According to the embodiments of this application, the connection methods of the antibody-coupled polypeptide molecules in Table 2 of the embodiments of this application are the same as those described in Ab1-P7-1 and Ab1-P7-2 above.

[0146] The antibody-conjugated polypeptide molecule of this application has good stability and can synergistically act on multiple targets to achieve effects such as fat reduction and muscle gain or improvement of blood lipids, while also improving blood sugar, weight and liver fat.

[0147] Composition In a third aspect of this application, a composition is provided comprising the antibody-coupled polypeptide molecule described in the second aspect.

[0148] According to embodiments of this application, the above composition may further include at least one of the following technical features: According to embodiments of this application, the composition further comprises pharmaceutically acceptable excipients.

[0149] According to embodiments of this application, pharmaceutically acceptable excipients refer to conventional pharmaceutical excipients in the pharmaceutical field, such as absorption enhancers, isotonic agents, stabilizers, regulators, etc.

[0150] According to embodiments of this application, a pharmaceutically acceptable carrier refers to a drug carrier conventional in the pharmaceutical field.

[0151] According to embodiments of this application, pharmaceutically acceptable media refers to conventional pharmaceutical media in the pharmaceutical field, such as solutions (e.g., water).

[0152] According to embodiments of this application, examples of suitable pharmaceutically acceptable carriers, excipients, and mediators are well known in the art. Pharmaceutical compositions comprising such carriers, excipients, and mediators can be formulated using known conventional methods.

[0153] In some alternative embodiments, the compositions of this application may also contain other active ingredients for therapeutic purposes.

[0154] The compositions of this application can be administered via various routes, such as oral or intravenous injection. Preferably, the compositions of this application are in solution form. Clinical dosing regimens are determined by the attending physician and clinical factors. As is known in the medical field, the dosage for any given patient depends on many factors, including patient size, body surface area, age, the drug to be administered, sex, time and route of administration, general health, and other concurrently administered medications. The compositions of this application can be administered topically or systemically. Preferably, they can be administered intravenously or subcutaneously. The compositions of this application can also be administered directly to the target site, for example, by targeted administration to internal or external target sites.

[0155] use In a fourth aspect of this application, the application provides the use of the antibody-drug conjugate molecule described in the second aspect or the composition described in the third aspect in the preparation of a medicament for the treatment or prevention of metabolic disorder-related diseases.

[0156] According to embodiments of this application, the metabolic disorder-related diseases include at least one of obesity, diabetes, dyslipidemia-related diseases, fatty liver disease, metabolic syndrome, and non-alcoholic fatty liver disease.

[0157] The terms “treatment” and “prevention” as used herein, and words derived therefrom, do not necessarily imply 100% or complete treatment or prevention. Rather, different degrees of treatment or prevention exist, and those skilled in the art will recognize that such treatment or prevention has potential benefit or therapeutic effect. Furthermore, the treatment or prevention provided in this application may include treatment or prevention of one or more diseases, such as cancer, or symptoms of a patient. Additionally, for the purposes of this document, “prevention” may encompass delaying the onset of a disease or its symptoms or the patient's condition.

[0158] Methods for preventing or treating metabolic disorders In another aspect of this application, a method for preventing or treating metabolic disorder-related diseases is proposed. According to embodiments of this application, the method includes administering to a subject a pharmaceutically acceptable amount of the antibody-drug conjugate molecule described in the second aspect or the composition described in the third aspect.

[0159] According to embodiments of this application, the metabolic disorder-related diseases include obesity, diabetes, dyslipidemia-related diseases, fatty liver disease, metabolic syndrome, and non-alcoholic fatty liver disease.

[0160] It should be noted that the "pharmaceuticalally acceptable amount" can vary depending on the mode of administration and the severity of the disease to be treated, and is preferably an effective amount. The selection of the pharmaceutically acceptable amount can be determined by those skilled in the art based on various factors (e.g., through clinical trials). These factors include, but are not limited to: the pharmacokinetic parameters of the active ingredient, such as bioavailability, metabolism, and half-life; the severity of the disease to be treated, the patient's weight, the patient's immune status, and the route of administration. For example, due to the urgency of the treatment condition, several separate doses may be administered daily, or the dose may be reduced proportionally.

[0161] In one alternative embodiment of this application, the pharmaceutically acceptable dose may be selected from the effective dose (or effective amount).

[0162] The antibody-drug conjugates or compositions of this application can be incorporated into pharmaceuticals suitable for use in various forms, such as liquids. Various routes of administration of the antibody-drug conjugates, compositions, or pharmaceuticals of this application are contemplated, including intravenous, intramuscular, and subcutaneous injection, but this application is not limited to these exemplified routes of administration.

[0163] The amino acid sequence of this application is shown in the table below:

[0164] The following will explain the solution of this application with reference to embodiments. Those skilled in the art will understand that the following embodiments are for illustrative purposes only and should not be considered as limiting the scope of this application. Where specific techniques or conditions are not specified in the embodiments, they are performed according to the techniques or conditions described in the literature in the art or according to the product instructions. Reagents or instruments whose manufacturers are not specified are all conventional products that can be obtained commercially.

[0165] Example 1: Solid-phase synthesis of GLP-1 receptor agonist peptide molecules containing linkers 1) Place the resin in a 150ml reactor and soak it in 50ml of dichloromethane (DCM) for 2 hours. Wash the resin with dimethylformamide (DMF) and then dry it. Repeat this process four times, drying the resin completely. Weigh out the first amino acid (protected) at the C-terminus of Fmoc, DCM, and N,N-diisopropylethylamine (DIEA) and add them to the reactor. Then place the reactor in a shaker at 30°C and react for 2 hours. Block the reaction with methanol solution (methanol:DIEA:DCM = 1:1:2) for half an hour, then wash four times with DMF and dry it completely. Add 20% piperidine solution to the reactor to remove the Fmoc protecting group. After deprotection, wash four times with DMF and then dry it completely.

[0166] 2) Weigh out the second amino acid (protected) at the C-terminus of Fmoc, 1-hydroxybenzotriazole (HOBT), and N,N-diisopropylcarbodiimide (DIC) and add them to the reactor. Then, place the reactor in a shaker at 30°C and react for 1 hour. Take a small amount of resin for testing using the ninhydrin method. If the resin is colored, it indicates incomplete condensation, and the reaction should continue. After the reaction is complete, wash the resin four times with DMF and then dry it. Add a certain amount of 20% piperidine (piperidine / DMF = 1:4) to the reactor and shake it on a decolorizing shaker for 20 minutes to remove the Fmoc protecting group from the resin. After deprotection, wash the resin four times with DMF and then dry it to check whether the protection has been removed.

[0167] 3) Link the amino acids sequentially according to the steps. Part of the Lys group (located at the C-terminus of the polypeptide) has its ε-amino group coupled to 2-bromoacetic acid, while another part of the Lys group (located in the middle of the polypeptide backbone) is linked to a linker. Protect the N-terminal amino group with Boc anhydride. Remove all protecting groups from the polypeptide using a cleavage reagent and cleave it from the resin for purification.

[0168] 4) The target peptides were purified by reversed-phase liquid chromatography (RP-HPLC) to separate impurities. The collected target peptides were lyophilized into powder and sent for QC quality control and purity testing via mass spectrometry. HPLC analysis showed that the purity of all peptides was greater than 95%. Mass spectrometry analysis confirmed that the molecular weights of the peptides were consistent with the theoretical molecular weights. The peptide compounds are shown in Table 1.

[0169] Table 1: GLP-1 receptor agonist peptide molecules containing linkers

[0170] G GGGSG GGGSG GGGSK(BrAc)-NH2 is the linker, where BrAc is -C(O)-CH2-Br. The Br in BrAc subsequently undergoes a substitution reaction with the -SH side chain of cysteine ​​in the targeting antibody or its functional fragment. -NH2 indicates that the hydroxyl group of amino acid K in GGGGSG GGGSG GGGSK(BrAc)-NH2 is replaced by -NH2.

[0171] Example 2: Construction and expression of antibody vectors 1) Molecular cloning methods were used to amplify the heavy chain variable region and light chain variable region fragments using a phage plasmid backbone. The variable region fragments were then constructed into backbone vectors containing the corresponding IgG subtype constant regions using the infusion method. After the sequence was verified to be correct, the plasmid vectors were extracted using an endotoxin-free plasmid extraction kit (OMEGA) and stored at -20℃.

[0172] 2) After resuscitating and passaged 293F cells, when the density reached approximately 3*10^6 cells / mL, the cells were transfected using PEI 40K for transient expression. The vector-to-PEI 40K mass ratio was 1:4, and the final vector concentration was 1ug / ml. On the second day after transfection, OPM-293-Profeed and glucose were added to maintain the growth of transfected cells. The cell culture medium was harvested when the cell viability dropped to approximately 60%.

[0173] 3) Centrifuge the cell culture medium to collect the supernatant, and filter it through a 0.45µm filter to remove residual cell debris. Purify the collected cell culture medium using a Protein A chromatography column, collecting the target peak. Further purification can be performed using cation exchange chromatography as needed. The protein is finally replaced by dialysis with 0.02M PBS. Samples are quantified using a NanoDrop 2000 / 2000c Spectrophotometer. Samples are analyzed by 12% SDS-PAGE electrophoresis, showing a single band. The precise molecular weight of the fusion protein is determined by mass spectrometry, and it is basically consistent with the theoretical molecular weight.

[0174] Example 3: Preparation of antibody-conjugated peptides 1) Take an appropriate amount of antibody solution, replace the buffer system with 40 mM HEPES (pH 8.0), and then add an appropriate concentration of cystamine and cysteamine. React at room temperature for 15-20 hours.

[0175] 2) Cation exchange was used to elute the bound antibody with elution buffer (20 mM PB, pH 7.5, 300 mM NaCl).

[0176] 3) Replace the sample solvent with a 0.01M sodium acetate buffer system containing 9% sucrose.

[0177] 4) Take an appropriate amount of antibody solution, add tris(2-carboxyethyl)phosphine (TCEP), the molar concentration of which is 3-5 times the molar concentration of the antibody, and reduce at room temperature for 30-120 minutes.

[0178] 5) Use cation exchange to elute the bound antibody with elution buffer (20 mM PB, pH 7.5, 300 mM NaCl). Then dilute with an equal volume of buffer (20 mM PB, pH 7.5) to adjust the sample buffer to 20 mM PB, pH 7.5, 150 mM NaCl, and finally determine the antibody concentration.

[0179] 6) Add EDTA (final concentration 2mM) and dehydroascorbic acid (DHAA) to the reduced antibody solution, with a molar concentration 10 times that of the antibody. Perform oxidation at room temperature for 90-180 minutes, and monitor the DHAA oxidation results using LCMS.

[0180] 7) Without removing DHAA, directly add 5-10 units of bromoacetyl-GLP-1 peptide and incubate at room temperature for 15-20 hours. Monitor the conjugation effect between the peptide and the antibody using LCMS.

[0181] 8) The antibody-conjugated peptide was purified by cation exchange, eluted and collected, and finally replaced with a buffer system of 20 mMPB, pH 7.5, 150 mM NaCl, and its concentration was measured.

[0182] Example 4: Determination of in vitro activity of peptide-conjugated antibodies before and after conjugation The sample, human GLP-1, human GCG, and human GIP were used to treat HEK293 cells overexpressing GLP-1R, GCGR, and GIPR, respectively. The cAMP produced by the recipient cells was detected using a cAMP assay kit (Cisbio, 62AM6PEC), a dose-response curve was established, and the EC50 was calculated. The specific steps are as follows: 1) Prepare the Assay buffer: Take the complete culture medium (DMEM medium + 10% FBS), and add 4 / 1000 of 500mM IBMX stock solution, cAMP-d2 working solution and anti-cAMP-crytate working solution, etc., and prepare according to the kit instructions. 2) Dilute the test sample and human control peptides GLP-1, GCG and GIP to an initial concentration of 500 nM stock solution, and then add 20 μL to 80 μL Assay buffer (diluted 5 times) in a gradient dilution, including a total of 8 compound gradients. 3) Cell suspension preparation: After removing HEK293-GLP-1R, HEK293-GCGR, and HEK293-GIPR cells from the liquid nitrogen container, immediately incubate them at 37°C for 1.5 min until completely thawed. Then, in a clean bench, add the cells dropwise to a 15 mL centrifuge tube containing 8 mL of warm culture medium. Centrifuge at 900 rpm for 5 min, discard the supernatant, and resuspend the cells in 1 mL of complete culture medium (pipe-pipette 15 times). Immediately take 20 μL of the suspension and mix it with an equal volume of trypan blue. Take 20 μL to count the number of viable cells, and then dilute the cells to 4 x 10^5 cells / mL. 4) Divide the 384-well plate into regions for GLP-1R cells, GCGR cells, and GIPR cells. Using a 12-channel adjustable dispenser, add 5 μL of cell suspension to each well of the corresponding region. Then, using the same 12-channel adjustable dispenser, add 5 μL of serially diluted test sample and positive control to each well of the corresponding cell type (two replicates for samples of the same concentration). For the negative control, add 10 μL of assay buffer per well. Set up three wells for each 384-well plate. Cover with a white sealing film and incubate at 37°C for half an hour. 5) Before use, dilute the cAMP-d2 working solution and anti-cAMP-crytate working solution 20 times with the lysis buffer in the kit, and mix them 1:1 to prepare a cAMP detection reagent mixture solution. Add 10 μL of cAMP detection reagent mixture solution to each well of the sample group, and add 5 μL of lysis buffer and 5 μL of diluted anti-cAMP-crytate working solution to each well of the negative control. Cover with a white cap and incubate at room temperature in the dark for 1 hour. 6) Fluorescence values ​​at 665 nm and 620 nm were detected using a multi-functional microplate reader. After data processing, the signal ratio and sample concentration were nonlinearly fitted using a four-parameter equation in GraphPadPrism 6 to obtain the EC50 value. The specific results are shown in Table 2.

[0183] Table 2: Cellular activity of antibody-conjugated peptides targeting human GLP-1 / GIP / GCG receptors

[0184] Example 5: Pharmacokinetic (PK) Study of Humanized FcRn Mice Thirty male humanized FcRn mice were divided into 10 groups of 3 mice each. Each compound was administered via a single subcutaneous injection at a dose of 5 mg / kg, with a volume of 5 mL / kg. Whole blood samples were collected at 2 h, 5 h, 8 h, 24 h, 48 h, 72 h, 96 h, 120 h, 144 h, 216 h, 336 h, 408 h, 504 h, 600 h, and 672 h post-administration. The collected whole blood samples were placed in anticoagulant tubes containing EDTA-K2, placed on wet ice, and centrifuged at 4ºC, 12000 rpm for 2 min within 1 h to separate the plasma. Store at 10ºC until testing.

[0185] Blood drug concentration assay: The concentrations of intact antibody-coupled peptide (ICP) molecules and total antibodies in blood samples are determined using the ELISA method. Intact ICP refers to antibodies still coupled with peptides; total antibodies refer to both peptide-coupled antibodies and antibodies without peptides.

[0186] The ELISA testing steps are as follows: 1) Coating: Dilute the capture antibody (or antigen) 1000-2000 times with 0.01M PBS, add 100 uL / well to each microplate, gently shake, and incubate overnight at 4°C.

[0187] 2) Sealing: Wash the plate 3 times, add 200 μL of 5% skim milk powder to each well, and seal at 25℃ and 400 rpm for about 2 hours.

[0188] 3) Sample preparation: Standard curve and QC samples were prepared by diluting mouse blank plasma.

[0189] 4) Sample dilution: Dilute the standard curve sample, QC sample, blank sample, and test sample with 1% BSAT at a ratio of 1:50 or 1:100. The DQC and test samples are further diluted with 1% BSAT containing blank plasma.

[0190] 5) Wash the plate: Wash the plate 3 times, add 100 μL of the above sample to each well, and incubate at 25℃ and 400 rpm for about 1.5 h.

[0191] 6) Add detection antibody: 100 μL / well 1:20000 diluted goat anti-human IgG-HRP (diluted with 1% BSAT), incubate at 25°C and 400 rpm for about 1 h.

[0192] 7) Add substrate: Wash the plate 3 times, add 100 μL of TMB substrate solution to each well, and incubate at 25℃ and 400 rpm for 5-20 minutes for color development. 8) Add stop solution: 50 μL / well stop solution.

[0193] 9) Detection: Absorbance is detected at 450 nm.

[0194] 10) Data processing: The pharmacokinetic parameters after drug administration were calculated using the non-compartmental model of WinNonlin 6.3 software, and the results are shown in Table 3.

[0195] In a humanized FcRn mouse model, the antibody-drug conjugates Ab1-P7-1 and Ab1-P7-2 of this application both exhibited excellent pharmacokinetic characteristics. They were well absorbed after subcutaneous administration, rapidly reaching high plasma concentrations, and both total molecular weight and intact active molecules showed high systemic exposure. Ab1-P7-2, in particular, exhibited higher intact molecular weight exposure and a longer half-life, demonstrating superior drug-like potential in terms of in vivo stability and long-lasting efficacy.

[0196] Table 3. Pharmacokinetic (PK) parameters of antibody-conjugated peptides in humanized FcRn mice

[0197] Example 6: Pharmacodynamic study in diet-induced obese C57BL / 6J mice (DIO) This embodiment evaluates the effects of Ab1-P7-1 and Ab1-P7-2 on body weight, muscle mass, and fat in DIO mice.

[0198] Experimental Methods: C57BL / 6J mice were randomly divided into a normal group and a model group at five weeks of age. The normal group was fed a standard maintenance diet, while the other groups were fed a high-fat diet (D12492). Body weight and food intake were monitored every 3 weeks. After 16 weeks of feeding, the differences between the two groups were statistically significant. The normal group was further divided into a Control group, and the successfully modeled mice were divided into Vehicle group, Tirzepatide group, positive GYM329+Tirzepatide group, Ab1+Tirzepatide group, Ab1-P7-1 group, and Ab1-P7-2 group. The Control group and Vehicle group were given the corresponding solvents, while the other groups were given the corresponding test products. All groups were administered the medication subcutaneously. The Tirzepatide dose was 15 nmol / kg / 3 quarterly doses, and the doses for positive GYM329, Ab1, Ab1-P7-1, and Ab1-P7-2 were 30 nmol / kg / quarterly doses. Administration lasted for 4 weeks. During the entire administration period, the Control group was fed a standard diet, while the other groups were fed a high-fat diet (D12492). Body composition was measured weekly using a small animal body composition analyzer (McGrey AccuFat-1050). Blood samples were collected on day 28 of administration, centrifuged at 12,000 g for 3 minutes at 4°C, and plasma was separated and stored at -80°C for analysis.

[0199] Experimental results: After 4 weeks of continuous administration, in terms of weight, all treatment groups showed a significant decrease compared to the Vehicle group. Compared to Tirzepatide, Ab1-P7-1 and Ab1-P7-2 showed a further significant decrease in weight. Figure 1 A-1B is shown in Table 4.

[0200] Regarding food intake, all treatment groups showed suppressed food intake compared to the Vehicle group, such as... Figure 1 As shown in C.

[0201] Considering both muscle mass and rate of muscle change, there were no statistically significant differences among the drug-treated groups compared to the Vehicle group. However, compared to Tirzepatide, Ab1-P7-1 was significantly increased. Figure 2 A-2B and Table 5-6.

[0202] Considering both fat mass and fat change rate, all treatment groups showed significant decreases compared to the Vehicle group. Compared to the Tirzepatide group, both Ab1-P7-1 and Ab1-P7-2 showed further significant decreases in fat mass. Figure 2 C-2D and Table 7-8).

[0203] In summary, both Ab1-P7-1 and Ab1-P7-2 can reduce fat while preventing muscle loss, ultimately bringing the body's muscle and fat content levels closer to the Control group.

[0204] Table 4: Effect on body weight of DIO mice

[0205] Note: The lowercase letter 'a' in the same column indicates a significant difference compared to the Vehicle group (P<0.05).

[0206] Table 5: Effects on muscle tissue in DIO mice

[0207] Note: The lowercase letter 'b' in the same column indicates a significant difference compared to the Tirzepatide group (P<0.05).

[0208] Table 6: Effects on muscle change rate in DIO mice

[0209] Note: The lowercase letter 'b' in the same column indicates a significant difference compared to the Tirzepatide group (P<0.05).

[0210] Table 7: Effects on fat in DIO mice

[0211] Note: The lowercase letter 'a' in the same column indicates a significant difference compared to the Vehicle group (P<0.05), and the lowercase letter 'b' in the same column indicates a significant difference compared to the Tirzepatide group (P<0.05).

[0212] Table 8: Effect on the rate of change in fat in DIO mice

[0213] Note: Lowercase letters in the same column indicate significant differences compared to the Vehicle group (P<0.05), and lowercase letters b in the same column indicate significant differences compared to the Tirzepatide group (P<0.05).

[0214] Example 7: Pharmacodynamic study in diet-induced obese C57BL / 6J mice (DIO) This embodiment evaluates the effects of Ab2, Iso-P5 (isotype antibody conjugated with P5 peptide), Ab2-P5, Ab2-P11, Ab2-P41, and Ab2-P43 on body weight, muscle mass, and fat in DIO mice.

[0215] Experimental Methods: C57BL / 6J mice were randomly divided into a normal group and a model group at five weeks of age. The normal group was fed a standard maintenance diet, while the other groups were fed a high-fat diet (D12492). Body weight and food intake were monitored every 3 weeks. After 16 weeks of feeding, the differences between the two groups were statistically significant. The normal group was further divided into a Control group, and the successfully modeled mice were divided into Vehicle, Ab2, Iso-P5, Ab2-P5, Ab2-P11, Ab2-P41, and Ab2-P43 groups. The Control and Vehicle groups were given the corresponding solvents, while the other groups were given the corresponding test products. All groups were administered the drugs subcutaneously. The Iso-P5 dose was 30 nmol / kg / 3 QD, and the dose for the other groups was 60 nmol / kg / QW. Administration lasted for 4 weeks. During the entire administration period, the Control group was fed a standard diet, while the other groups were fed a high-fat diet (D12492). Blood samples were collected on day 28 of drug administration. The plasma was separated by centrifugation at 12,000 g for 3 minutes at 4°C and stored at -80°C for analysis. Subsequently, fat (subcutaneous, epididymal, perirenal) and muscle (quadriceps, tibialis anterior, gastrocnemius, soleus) were collected and weighed.

[0216] Experimental results: After 4 weeks of continuous administration, compared with Vehicle, except for Ab2, the body weight of mice in all treatment groups decreased significantly, such as... Figure 3 See Table 9.

[0217] Regarding muscle mass, compared to the Vehicle group, except for Iso-P5 which showed no statistically significant difference, all other treatment groups showed a significant increase in muscle mass, such as... Figure 4 A and Table 10).

[0218] Regarding fat, compared to the Vehicle group, all other treatment groups showed a significant decrease in fat content, except for Ab2, which did not show a significant decrease. Figure 4 B and Table 11).

[0219] In summary, Ab2-P5, Ab2-P11, Ab2-P41, Ab2-P43, and Ab2-PNN02 can all reduce fat while preventing muscle loss.

[0220] Table 9: Effects on body weight of DIO mice

[0221] Note: The lowercase letter 'a' in the same column indicates a significant difference compared to the Vehicle group (P<0.05).

[0222] Table 10: Effects on muscle tissue in DIO mice

[0223] Note: The lowercase letter 'a' in the same column indicates a significant difference compared to the Vehicle group (P<0.05).

[0224] Table 11: Effects on fat in DIO mice

[0225] Note: Lowercase letters in the same column indicate significant differences compared to the Vehicle group (P<0.05).

[0226] In the description of this specification, the references to terms such as "one embodiment," "some embodiments," "example," "specific example," or "some examples," etc., indicate that a specific feature, structure, material, or characteristic described in connection with that embodiment or example is included in at least one embodiment or example of this application. In this specification, the illustrative expressions of the above terms do not necessarily refer to the same embodiment or example. Furthermore, the specific features, structures, materials, or characteristics described may be combined in any suitable manner in one or more embodiments or examples. Moreover, without contradiction, those skilled in the art can combine and integrate the different embodiments or examples described in this specification, as well as the features of different embodiments or examples.

[0227] Although embodiments of this application have been shown and described above, it is understood that the above embodiments are exemplary and should not be construed as limiting this application. Those skilled in the art can make changes, modifications, substitutions and variations to the above embodiments within the scope of this application.

Claims

1. A GLP-1 receptor agonist polypeptide molecule, characterized in that, It has the structure shown in equation (I): X1-Aib-X3GTX6TSDYSIX 13 LDX 16 X 17 X 18 X 19 X 20 X 21 FX 23 X 24 X 25 LX 27 X 28 GGPSSGAPPPS(I); in: X1 is either Y or H; X3 is either Q or E; X6 is αMeF(2F) or F; X 13 For L, αMeL, or Aib; X 16 For E, K, or R; X 17 For K, I, or Q; X 18 For R or A; X 19 For Q or A; X 20 For Aib, R, K, or Q; X 21 It can be A or E; X 23 For I or V; X 24 For E or Q; X 25 For Y or W; X 27 For L or I; X 28 It can be A or E.

2. The GLP-1 receptor agonist polypeptide molecule according to claim 1, characterized in that, The GLP-1 receptor agonist polypeptide molecule is a GLP-1 / GCG / GIP triad receptor agonist polypeptide molecule, and the structure shown in formula (I) is as shown in formula (Ia): X1-Aib-QGTX6TSDYSIX 13 LDX 16 X 17 X 18 QX 20 X 21 IN WHAT 25 LX 27 X 28 GGPSSGAPPPS(Ia); in: X1 is either Y or H; X6 is αMeF(2F) or F; X 13 For L or αMeL; X 16 It can be E or K; X 17 For K or I; X 18 For R or A; X 20 For Aib or R; X 21 It can be A or E; X 25 For Y or W; X 27 For L or I; X 28 It can be A or E; Optionally, the GLP-1 receptor agonist polypeptide molecule is a GLP-1 / GIP dual receptor agonist polypeptide molecule, and the structure shown in formula (I) is as shown in formula (Ib): Y-Aib-EGTFTSDYSIX 13 LDX 16 X 17 AX 19 X 20 X 21 FX 23 X 24 WLX 27 AGGPSSGAPPPS(Ib); in: X 13 For L or Aib; X 16 For E, K, or R; X 17 For I or Q; X 19 For Q or A; X 20 For R, K, or Q; X 21 It can be A or E; X 23 For I or V; X 24 For E or Q; X 27 For L or I; Optionally, the GLP-1 receptor agonist polypeptide molecule has at least one of the following structural formulas: Y-Aib-QGT αMeF(2F)-TSDY SILLD EKRQ-Aib AFIEY LLAGG PSSGA PPPS; Y-Aib-QGT αMeF(2F)-TSDY SI-aMeL-LD EKRQ-Aib AFIEY LLAGG PSSGA PPPS; Y-Aib-QGT αMeF(2F)TSDY SILLD KIRQ-Aib EFIEW LLAGG PSSGA PPPS; Y-Aib-QGT αMeF(2F)-TSDY SILLD KIAQR EFIEW LLAGG PSSGA PPPS; Y-Aib-QGT FTSDY SILLD KIAQ-Aib EFIEW LIAGG PSSGA PPPS; Y-Aib-QGT αMeF(2F)-TSDY SILLD KIAQ-Aib EFIEW LLAGG PSSGA PPPS; H-Aib-QGT FTSDY SILLD KIAQR EFIEW LLAGG PSSGA PPPS; Y-Aib-QGT αMeF(2F)-TSDY SILLD KKAQ-Aib EFIEW LLAGG PSSGA PPPS; Y-Aib-QGT FTSDY SILLD KIAQ-Aib EFIEW LLAGG PSSGA PPPS; Y-Aib-QGT FTSDY SILLD KIAQR EFIEW LLAGG PSSGA PPPS; Y-Aib-QGT FTSDY SILLD KIAQR EFIEW LLEGG PSSGA PPPS; Y-Aib-QGT FTSDY SILLD KIAQR AFIEY LLAGG PSSGA PPPS; Y-Aib-QGT FTSDY SILLD KIAQR AFIEY LLEGG PSSGA PPPS; Y-Aib-QGT FTSDY SILLD KIAQR EFIEW LIAGG PSSGA PPPS; Y-Aib-EGT FTSDY SILLD RIAQR EFIEW LLAGG PSSGA PPPS; Y-Aib-EGT FTSDY SI-Aib-LD KIAQK AFVQW LIAGG PSSGA PPPS; Y-Aib-EGT FTSDY SILLD EQAAR EFIEW LLAGG PSSGA PPPS; Y-Aib-EGT FTSDY SILLD KIAQR EFIEW LLAGG PSSGA PPPS; Y-Aib-EGT FTSDY SILLD KIAQQ EFIEW LLAGG PSSGA PPPS.

3. An antibody-conjugated polypeptide molecule, characterized in that, This includes targeting antibodies or their functional fragments, as well as GLP-1 receptor agonists.

4. The antibody-conjugated polypeptide molecule according to claim 3, characterized in that, The targeting antibody or its functional fragment is conjugated to a GLP-1 receptor agonist; Optionally, the targeting antibody or its functional fragment is conjugated to a GLP-1 receptor agonist via a linker; Optionally, the antibody-conjugated polypeptide molecule has the structure shown in formula (II): XZU(II); in: U represents the targeting antibody or its functional fragment; Z stands for connector; X is a GLP-1 receptor agonist.

5. The antibody-conjugated polypeptide molecule according to claim 4, characterized in that, The antibody-conjugated polypeptide molecule satisfies one or more of the following conditions: i) The GLP-1 receptor agonist is selected from one of the following: a GLP-1 monoreceptor agonist, a GLP-1 / GIP dual-receptor agonist, or a GLP-1 / GCG / GIP triple-receptor agonist; Optionally, the GLP-1 receptor agonist is a GLP-1 / GIP dual receptor agonist; Optionally, the GLP-1 receptor agonist is a GLP-1 / GCG / GIP triad receptor agonist; Optionally, the GLP-1 receptor agonist is selected from at least one of the GLP-1 receptor agonist peptide molecules of claim 1; ii) The targeting antibody or its functional fragment targets one of the following: activin type 2 receptor (ActRII), growth differentiation factor (GDF-8), leptin receptor (LepR), PCSK9, and cannabinoid receptor (CB1); Optionally, the targeting antibody or its functional fragment targets Pro-GDF-8, Latent GDF-8, or mature GDF-8; Optionally, the targeting antibody or a functional fragment thereof is used to target Latent GDF-8; Optionally, the targeted antibody is selected from at least one of full-length monoclonal antibodies, Fab antibodies, Fab' antibodies, F(ab')2 antibodies, Fv antibodies, single-chain antibodies, dsFv antibodies, and nanobodies; Optionally, the targeting antibody or its functional fragment is selected from full-length monoclonal antibodies or Fab antibodies; iii) The connector contains a GS linker peptide; Optionally, the GS linker peptide comprises at least one of the amino acid sequences shown in (G3S)2, (G4S)2, (G3S)3, (G4S)3, (G3S)4, (G4S)4, (G3S)5, (G4S)5, (G3S)6, and (G4S)6. Optionally, the connector further includes an amino acid K and / or an acetyl unit (-C(O)-CH2-). Optionally, the GS linker peptide and the acetyl unit are respectively linked to two amino groups of amino acid K, and the * end of the acetyl unit (-C(O)-CH2-*) is used to link the targeting antibody or its functional fragment; Optionally, the linker is selected from GS linker peptide-amino acid K-acetyl unit; Optionally, the hydroxyl group on the carboxyl group of the amino acid K is replaced with -NH2; Optionally, the linker is selected from GGGGSGGGGSGGGGSK(-C(O)-CH2-*)-NH2, the * end is linked to the targeting antibody or its functional fragment, the N end of GGGGSGGGGSGGGGS in the linker is linked to the GLP-1 receptor agonist, and -NH2 indicates that the hydroxyl group of amino acid K is replaced by -NH2; iv) One end of the connector is connected to the C-terminus of a GLP-1 receptor agonist or any lysine residue on the polypeptide chain of a GLP-1 receptor agonist, and the other end is connected to a targeting antibody or its functional fragment. v) The targeting antibody or its functional fragment has one or more coupling sites, and the targeting antibody or its functional fragment is coupled to the GLP-1 receptor agonist through the coupling sites; Optionally, the targeting antibody or its functional fragment has two coupling sites, each of which is coupled to a GLP-1 receptor agonist. Optionally, the coupling site is selected from cysteine ​​or substituted cysteine; Optionally, the GLP-1 receptor agonist is coupled to the side chain of a cysteine ​​or a substituted cysteine ​​at the coupling site.

6. The antibody-conjugated polypeptide molecule according to claim 4, characterized in that, The targeting antibody or its functional fragments include: complementation-determining regions HCDR1, HCDR2 and HCDR3 of the heavy chain variable region, and complementation-determining regions LCDR1, LCDR2 and LCDR3 of the light chain variable region; The HCDR1, HCDR2 and HCDR3 have the HCDR1, HCDR2 and HCDR3 defined in the heavy chain variable region as shown in the amino acid sequence of SEQ ID NO:13 or SEQ ID NO:15; The LCDR1, LCDR2 and LCDR3 have LHCDR1, LCDR2 and LCDR3 defined in the heavy chain variable region as shown in the amino acid sequence of SEQ ID NO:14 or SEQ ID NO:16; Optionally, HCDR1, HCDR2, and HCDR3 have HCDR1, HCDR2, and HCDR3 defined in the heavy chain variable region as shown in the amino acid sequence of SEQ ID NO:13, and LCDR1, LCDR2, and LCDR3 have LHCDR1, LCDR2, and LCDR3 defined in the heavy chain variable region as shown in the amino acid sequence of SEQ ID NO:14; or The HCDR1, HCDR2 and HCDR3 have HCDR1, HCDR2 and HCDR3 defined in the heavy chain variable region as shown in the amino acid sequence of SEQ ID NO:15, and the LCDR1, LCDR2 and LCDR3 have LHCDR1, LCDR2 and LCDR3 defined in the heavy chain variable region as shown in the amino acid sequence of SEQ ID NO:

16. Optionally, HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 are amino acid sequences obtained using the definitions of IMGT, Kabat, Chothia, AbM, or Contact. Optionally, the targeting antibody or its functional fragment includes: complementation-determining regions HCDR1, HCDR2 and HCDR3 of the heavy chain variable region, and complementation-determining regions LCDR1, LCDR2 and LCDR3 of the light chain variable region; The HCDR1 has an amino acid sequence as shown in SEQ ID NO:1 or 7, or an amino acid sequence that is at least 60% identical to SEQ ID NO:1 or 7, or an amino acid sequence that differs from SEQ ID NO:1 or 7 by 3, 2 or 1 amino acids. The HCDR2 has an amino acid sequence as shown in SEQ ID NO:2 or 8, or an amino acid sequence that is at least 80% identical to SEQ ID NO:2 or 8, or an amino acid sequence that differs from SEQ ID NO:2 or 8 by 3, 2 or 1 amino acids. The HCDR3 has an amino acid sequence as shown in SEQ ID NO:3 or 9, or an amino acid sequence that is at least 80% identical to SEQ ID NO:3 or 9, or an amino acid sequence that differs from SEQ ID NO:3 or 9 by 3, 2 or 1 amino acids. The LCDR1 has an amino acid sequence as shown in SEQ ID NO:4 or 10, or an amino acid sequence that is at least 60% identical to SEQ ID NO:4 or 10, or an amino acid sequence that differs from SEQ ID NO:4 or 10 by 3, 2 or 1 amino acids. The LCDR2 has an amino acid sequence as shown in SEQ ID NO:5 or 11, or an amino acid sequence that is at least 80% identical to SEQ ID NO:5 or 11, or an amino acid sequence that differs from SEQ ID NO:5 or 11 by 3, 2 or 1 amino acids. The LCDR3 has an amino acid sequence as shown in SEQ ID NO:6 or 12, or an amino acid sequence that is at least 80% identical to SEQ ID NO:6 or 12, or an amino acid sequence that differs from SEQ ID NO:6 or 12 by 3, 2 or 1 amino acids. Optionally, HCDR1, HCDR2, and HCDR3 are selected from any one of the following: a1) The HCDR1 has an amino acid sequence as shown in SEQ ID NO:1, or an amino acid sequence that is at least 60% identical to SEQ ID NO:1, or an amino acid sequence that differs from SEQ ID NO:1 by 3, 2 or 1 amino acids. The HCDR2 has an amino acid sequence as shown in SEQ ID NO:2, or an amino acid sequence that is at least 80% identical to SEQ ID NO:2, or an amino acid sequence that differs from SEQ ID NO:2 by 3, 2 or 1 amino acids. The HCDR3 has an amino acid sequence as shown in SEQ ID NO:3, or an amino acid sequence that is at least 80% identical to SEQ ID NO:3, or an amino acid sequence that differs from SEQ ID NO:3 by 3, 2 or 1 amino acids. a2) The HCDR1 has an amino acid sequence as shown in SEQ ID NO:7, or an amino acid sequence that is at least 60% identical to SEQ ID NO:7, or an amino acid sequence that differs from SEQ ID NO:7 by 3, 2 or 1 amino acids; The HCDR2 has an amino acid sequence as shown in SEQ ID NO:8, or an amino acid sequence that is at least 80% identical to SEQ ID NO:8, or an amino acid sequence that differs from SEQ ID NO:8 by 3, 2 or 1 amino acids. The HCDR3 has an amino acid sequence as shown in SEQ ID NO:9, or an amino acid sequence that is at least 80% identical to SEQ ID NO:9, or an amino acid sequence that differs from SEQ ID NO:9 by 3, 2 or 1 amino acids. Optionally, LCDR1, LCDR2, and LCDR3 are selected from any one of the following: b1) The LCDR1 has an amino acid sequence as shown in SEQ ID NO:4, or an amino acid sequence that is at least 60% identical to SEQ ID NO:4, or an amino acid sequence that differs from SEQ ID NO:4 by 3, 2 or 1 amino acids. The LCDR2 has an amino acid sequence as shown in SEQ ID NO:5, or an amino acid sequence that is at least 80% identical to SEQ ID NO:5, or an amino acid sequence that differs from SEQ ID NO:5 by 3, 2 or 1 amino acids. The LCDR3 has an amino acid sequence as shown in SEQ ID NO:6, or an amino acid sequence that is at least 80% identical to SEQ ID NO:6, or an amino acid sequence that differs from SEQ ID NO:6 by 3, 2 or 1 amino acids. b2) The LCDR1 has an amino acid sequence as shown in SEQ ID NO:10, or an amino acid sequence that is at least 60% identical to SEQ ID NO:10, or an amino acid sequence that differs from SEQ ID NO:10 by 3, 2 or 1 amino acids. The LCDR2 has an amino acid sequence as shown in SEQ ID NO:11, or an amino acid sequence that is at least 80% identical to SEQ ID NO:11, or an amino acid sequence that differs from SEQ ID NO:11 by 3, 2 or 1 amino acids. The LCDR3 has an amino acid sequence as shown in SEQ ID NO:12, or an amino acid sequence that is at least 80% identical to SEQ ID NO:12, or an amino acid sequence that differs from SEQ ID NO:12 by 3, 2 or 1 amino acids. Optionally, HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 are selected from any one of the following: c1) The HCDR1 has an amino acid sequence as shown in SEQ ID NO:1, or an amino acid sequence that is at least 60% identical to SEQ ID NO:1, or an amino acid sequence that differs from SEQ ID NO:1 by 3, 2 or 1 amino acids. The HCDR2 has an amino acid sequence as shown in SEQ ID NO:2, or an amino acid sequence that is at least 80% identical to SEQ ID NO:2, or an amino acid sequence that differs from SEQ ID NO:2 by 3, 2 or 1 amino acids. The HCDR3 has an amino acid sequence as shown in SEQ ID NO:3, or an amino acid sequence that is at least 80% identical to SEQ ID NO:3, or an amino acid sequence that differs from SEQ ID NO:3 by 3, 2 or 1 amino acids. The LCDR1 has an amino acid sequence as shown in SEQ ID NO:4, or an amino acid sequence that is at least 60% identical to SEQ ID NO:4, or an amino acid sequence that differs from SEQ ID NO:4 by 3, 2 or 1 amino acids. The LCDR2 has an amino acid sequence as shown in SEQ ID NO:5, or an amino acid sequence that is at least 80% identical to SEQ ID NO:5, or an amino acid sequence that differs from SEQ ID NO:5 by 3, 2 or 1 amino acids. The LCDR3 has an amino acid sequence as shown in SEQ ID NO:6, or an amino acid sequence that is at least 80% identical to SEQ ID NO:6, or an amino acid sequence that differs from SEQ ID NO:6 by 3, 2 or 1 amino acids. c2) The HCDR1 has an amino acid sequence as shown in SEQ ID NO:7, or an amino acid sequence that is at least 60% identical to SEQ ID NO:7, or an amino acid sequence that differs from SEQ ID NO:7 by 3, 2 or 1 amino acids. The HCDR2 has an amino acid sequence as shown in SEQ ID NO:8, or an amino acid sequence that is at least 80% identical to SEQ ID NO:8, or an amino acid sequence that differs from SEQ ID NO:8 by 3, 2 or 1 amino acids. The HCDR3 has an amino acid sequence as shown in SEQ ID NO:9, or an amino acid sequence that is at least 80% identical to SEQ ID NO:9, or an amino acid sequence that differs from SEQ ID NO:9 by 3, 2 or 1 amino acids. The LCDR1 has an amino acid sequence as shown in SEQ ID NO:10, or an amino acid sequence that is at least 60% identical to SEQ ID NO:10, or an amino acid sequence that differs from SEQ ID NO:10 by 3, 2 or 1 amino acids. The LCDR2 has an amino acid sequence as shown in SEQ ID NO:11, or an amino acid sequence that is at least 80% identical to SEQ ID NO:11, or an amino acid sequence that differs from SEQ ID NO:11 by 3, 2 or 1 amino acids. The LCDR3 has an amino acid sequence as shown in SEQ ID NO:12, or an amino acid sequence that is at least 80% identical to SEQ ID NO:12, or an amino acid sequence that differs from SEQ ID NO:12 by 3, 2 or 1 amino acids. Optionally, HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 are selected from any one of the following: d1) The amino acid sequences of HCDR1, HCDR2 and HCDR3 are shown in SEQ ID NO:1, SEQ ID NO:2 and SEQ ID NO:3 respectively, and the amino acid sequences of LCDR1, LCDR2 and LCDR3 are shown in SEQ ID NO:4, SEQ ID NO:5 and SEQ ID NO:6 respectively. d2) The amino acid sequences of HCDR1, HCDR2 and HCDR3 are shown in SEQ ID NO:7, SEQ ID NO:8 and SEQ ID NO:9 respectively, and the amino acid sequences of LCDR1, LCDR2 and LCDR3 are shown in SEQ ID NO:10, SEQ ID NO:11 and SEQ ID NO:12 respectively. Optionally, the heavy chain variable region includes a heavy chain framework region, the heavy chain framework region being derived from at least one of a mouse antibody, a human antibody, a primate antibody, or a mutant thereof; and / or, the light chain variable region includes a light chain framework region, the light chain framework region being derived from at least one of a mouse antibody, a human antibody, a primate antibody, or a mutant thereof; Optionally, the heavy chain frame region includes HFR1, HFR2, HFR3 and HFR4, and / or the light chain frame region includes LFR1, LFR2, LFR3 and LFR4; Optionally, the heavy chain variable region has an amino acid sequence as shown in SEQ ID NO: 13 or 15, or an amino acid sequence that is at least 80% identical to SEQ ID NO: 13 or 15, or an amino acid sequence that differs from SEQ ID NO: 13 or 15 by 3, 2 or 1 amino acids. Optionally, the light chain variable region has an amino acid sequence as shown in SEQ ID NO: 14 or 16, or an amino acid sequence that is at least 80% identical to SEQ ID NO: 14 or 16, or an amino acid sequence that differs from SEQ ID NO: 14 or 16 by 3, 2 or 1 amino acids. Optionally, the heavy chain variable region has the amino acid sequence shown in SEQ ID NO:13, and the light chain variable region has the amino acid sequence shown in SEQ ID NO:14; or The heavy chain variable region has an amino acid sequence as shown in SEQ ID NO:15, and the light chain variable region has an amino acid sequence as shown in SEQ ID NO:16; Optionally, the targeting antibody or its functional fragment further includes a heavy chain constant region and / or a light chain constant region; Optionally, the constant region is derived from at least one of camel-derived antibodies, mouse-derived antibodies, human-derived antibodies, primate-derived antibodies, or mutants thereof; Optionally, the heavy chain constant region is selected from a portion or all of the heavy chain constant regions of one of IgG1, IgG2, IgG3, IgG4, IgA, IgM, IgE or IgD. Optionally, the heavy chain constant region is selected from one or more of the CH1, CH2, CH3 and CH4 regions.

7. The antibody-conjugated polypeptide molecule according to any one of claims 4 to 6, characterized in that, The heavy chain constant region of the targeting antibody or its functional fragment includes an Fc region and optionally a CH1 region; Optionally, the coupling site is located on the Fc region; Optionally, one end of the connector is connected to the C-terminus of a GLP-1 receptor agonist or any lysine residue on a GLP-1 receptor agonist, and the other end is connected to a cysteine ​​or a substituted cysteine ​​residue on the Fc region of the targeting antibody or its functional fragment. Optionally, the coupling site is selected from the 272nd position on the Fc region; Optionally, the 272nd position is a substituted cysteine.

8. An antibody-conjugated polypeptide molecule, characterized in that, It contains targeting antibodies or their functional fragments and GLP-1 receptor agonists. The antibody or its functional fragment targets one of the following: activin type 2 receptor (ActRII), growth differentiation factor (GDF-8), leptin receptor (LepR), PCSK9, and cannabinoid receptor (CB1); Optionally, the antibody or a functional fragment thereof targets GDF-8; Optionally, the antibody or its functional fragment targets Pro-GDF-8, Latent GDF-8, or mature GDF-8; Optionally, the antibody or a functional fragment thereof targets Latent GDF-8; Optionally, the GLP-1 receptor agonist is selected from the GLP-1 receptor agonist polypeptide molecule described in claim 1; Optionally, the Latent GDF-8 antibody includes: complementation-determining regions HCDR1, HCDR2 and HCDR3 of the heavy chain variable region, and complementation-determining regions LCDR1, LCDR2 and LCDR3 of the light chain variable region; Wherein, HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 are selected from any one of the following: d1) The amino acid sequences of HCDR1, HCDR2 and HCDR3 are shown in SEQ ID NO:1, SEQ ID NO:2 and SEQ ID NO:3 respectively, and the amino acid sequences of LCDR1, LCDR2 and LCDR3 are shown in SEQ ID NO:4, SEQ ID NO:5 and SEQ ID NO:6 respectively. d2) The amino acid sequences of HCDR1, HCDR2 and HCDR3 are shown in SEQ ID NO:7, SEQ ID NO:8 and SEQ ID NO:9 respectively, and the amino acid sequences of LCDR1, LCDR2 and LCDR3 are shown in SEQ ID NO:10, SEQ ID NO:11 and SEQ ID NO:12 respectively.

9. An antibody-conjugated polypeptide molecule, characterized in that, Contains a latent GDF-8 antibody and a GLP-1 receptor agonist; The GLP-1 receptor agonist is selected from the GLP-1 receptor agonist polypeptide molecule described in claim 1; The Latent GDF-8 antibody includes: complementation-determining regions HCDR1, HCDR2 and HCDR3 of the heavy chain variable region, and complementation-determining regions LCDR1, LCDR2 and LCDR3 of the light chain variable region; Wherein, HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 are selected from any one of the following: d1) The amino acid sequences of HCDR1, HCDR2 and HCDR3 are shown in SEQ ID NO:1, SEQ ID NO:2 and SEQ ID NO:3 respectively, and the amino acid sequences of LCDR1, LCDR2 and LCDR3 are shown in SEQ ID NO:4, SEQ ID NO:5 and SEQ ID NO:6 respectively. d2) The amino acid sequences of HCDR1, HCDR2 and HCDR3 are shown in SEQ ID NO:7, SEQ ID NO:8 and SEQ ID NO:9 respectively, and the amino acid sequences of LCDR1, LCDR2 and LCDR3 are shown in SEQ ID NO:10, SEQ ID NO:11 and SEQ ID NO:12 respectively.

10. An antibody-conjugated polypeptide molecule, characterized in that, The product comprises a latent GDF-8 antibody, a linker, and a GLP-1 receptor agonist; wherein the latent GDF-8 antibody and the GLP-1 receptor agonist comprise any one of the following groups: Wherein, -C(O)-CH2-* represents an acetyl unit, the * end in -C(O)-CH2-* is used to connect the Latent GDF-8 antibody, and -NH2 indicates that the hydroxyl group on the carboxyl group of the amino acid K connected to -C(O)-CH2-* is replaced by -NH2. The Latent GDF-8 antibody Ab1 includes complementarity-determining regions HCDR1, HCDR2, and HCDR3 of the heavy chain variable region, and complementarity-determining regions LCDR1, LCDR2, and LCDR3 of the light chain variable region. The amino acid sequences of HCDR1, HCDR2, and HCDR3 are shown in SEQ ID NO:1, SEQ ID NO:2, and SEQ ID NO:3, respectively, and the amino acid sequences of LCDR1, LCDR2, and LCDR3 are shown in SEQ ID NO:4, SEQ ID NO:5, and SEQ ID NO:6, respectively. The Latent GDF-8 antibody Ab1 includes complementarity-determining regions HCDR1, HCDR2, and HCDR3 of the heavy chain variable region, and complementarity-determining regions LCDR1, LCDR2, and LCDR3 of the light chain variable region. The amino acid sequences of HCDR1, HCDR2, and HCDR3 are shown in SEQ ID NO:7, SEQ ID NO:8, and SEQ ID NO:9, respectively, and the amino acid sequences of LCDR1, LCDR2, and LCDR3 are shown in SEQ ID NO:10, SEQ ID NO:11, and SEQ ID NO:12, respectively.

11. A composition, characterized in that, Includes the antibody-conjugated polypeptide molecule as described in any one of claims 1 to 10.

12. The composition according to claim 11, characterized in that, This further includes pharmaceutically acceptable excipients.

13. Use of the antibody-conjugated polypeptide molecule of any one of claims 1 to 10 or the composition of any one of claims 11 to 12 in the preparation of a medicament for the treatment or prevention of metabolic disorder-related diseases.

14. The use according to claim 13, characterized in that, The metabolic disorder-related diseases include at least one of obesity, diabetes, dyslipidemia-related diseases, fatty liver disease, metabolic syndrome, and non-alcoholic fatty liver disease.

Images