Cyclic rnas encoding fgf21 and uses thereof

Abstract

This application provides a circular RNA encoding FGF21 and its use in treating diseases such as obesity, diabetes, or metabolic disorder-associated fatty liver disease (MAFLD) or steatohepatitis (MASH).

Description

Technical Field

[0001] This application relates to a circular RNA encoding FGF21 and its use in disease treatment. Background Technology

[0002] Metabolic disorders, including type 2 diabetes (T2D), obesity, and metabolic dysfunction-associated fatty liver disease (MAFLD), have a global prevalence of approximately 25%, contributing to the global healthcare burden (Jamali, Zahra et al., Scientific reports. 2024). Semaglutide, a glucagon-like peptide-1 receptor agonist, has been approved for the treatment of type 2 diabetes (T2D) and weight loss. However, gastrointestinal problems and muscle loss are frequently observed during semaglutide treatment. Although the selective thyroid hormone receptor beta agonist resmetirom was first approved in 2024 for the treatment of non-cirrhotic non-alcoholic steatohepatitis (NASH) in adults, its clinical benefits appear contradictory in existing studies (Harrison, Stephen A et al., Lancet (London, England). 2019). Currently, fibroblast growth factor 21 (FGF-21) analogues have emerged as a promising new treatment option for metabolic diseases.

[0003] FGF-21 is an endocrine hormone belonging to the FGF superfamily. Primarily secreted by the liver and released into the bloodstream, FGF21 acts on adipose tissue, liver, skeletal muscle, and other distal tissues via the fibroblast growth factor receptor (FGFR) and the β-Klotho complex (Tezze, C., Romanello, V., & Sandri, M. Frontiers inphysiology, 2019). The FGF21 / FGFR / β-Klotho complex triggers the stepwise activation of the downstream ERK / MAPK signaling cascade, thereby regulating cell proliferation, differentiation, and metabolism. FGF21 analogues have been investigated for their potential to regulate dyslipidemia, insulin sensitivity, hepatic steatosis, and fibrosis, all of which are associated with the progression of type 2 diabetes, obesity, and MAFLD (Harrison, Stephen A et al., Journal of hepatology. 2024). Therefore, FGF21 holds therapeutic potential in various metabolic diseases.

[0004] The human FGF21 precursor protein consists of 208 amino acids, which matures into a secretory form containing 181 amino acids during maturation. This protein is primarily secreted by the liver and adipose tissue (DM Kilkenny, JV Rocheleau, Vitamins & Hormones, 2016). However, the utilization of natural FGF21 is limited due to its short half-life. The half-life of FGF21 in animal models has been reported to be approximately 0.5–2 hours (Kharitonenkov et al., Endocrinology, 2007; Xu et al., Am. J. Physiol. Endocrinol. Metab, 2009; Hecht et al., PLoS One, 2012). FGF21 is cleaved at the N-terminus by dipeptidyl peptidase IV (DPP-IV) and at the C-terminus by fibroblast activator protein (FAP) (Zhen, Eugene Y et al., The Biochemical journal, 2016). Several strategies have been applied to the molecular engineering of FGF21 and its analogues, such as PEGylated analogues and antibody-based fusion proteins (Tillman EJ, Rolph T. Front Endocrinol (Lausanne), 2020). Despite these advances, anti-PEG antibodies, vacuolation caused by PEGylated molecules, and insufficient stability remain challenges to its development.

[0005] Therefore, there is a need in this field to produce better FGF21 analogs to treat metabolic diseases. Summary of the Invention

[0006] This application relates to a novel circular RNA encoding FGF21 or a variant thereof, which has improved pharmaceutical properties compared to recombinant proteins.

[0007] In one aspect, this application provides a circular RNA comprising a regulatory element and an expression element, wherein the expression element comprises a polynucleotide encoding FGF21 or a variant thereof.

[0008] In one aspect, this application provides a composition comprising the circular RNA disclosed herein, wherein the composition further comprises a pharmaceutically acceptable excipient.

[0009] In one aspect, this application provides a method for treating a subject's disease, comprising administering to the subject a therapeutically effective amount of the circular RNA or composition disclosed in this application.

[0010] In one aspect, this application provides the use of the circular RNA or composition disclosed herein in the preparation of a medicament for treating a subject’s disease.

[0011] In one aspect, this application provides the circular RNA or composition disclosed herein for use in treating a subject's disease.

[0012] In one aspect, this application provides a method for reducing the weight of a subject without reducing the subject's lean body mass, the method comprising administering to the subject a therapeutically effective amount of the circular RNA or composition disclosed in this application.

[0013] In one aspect, this application provides a method for reducing a subject's fat without reducing the subject's lean body mass, the method comprising administering to the subject a therapeutically effective amount of the circular RNA or composition disclosed in this application.

[0014] In one aspect, this application provides a method for reducing the weight of a subject, the method comprising administering to the subject a therapeutically effective amount of the circular RNA or composition disclosed in this application.

[0015] In one aspect, this application provides a method for lowering total cholesterol, low-density lipoprotein cholesterol (LDL-C), or blood glucose in a subject, the method comprising administering to the subject a therapeutically effective amount of the circular RNA or composition disclosed in this application.

[0016] In one aspect, this application provides a method for reducing hepatic steatosis in a subject, the method comprising administering to the subject a therapeutically effective amount of the circular RNA or composition disclosed in this application. Attached Figure Description

[0017] Figure 1 This is an overview diagram of plasmids used to produce circular RNA.

[0018] Figure 2 Urea-polyacrylamide gel electrophoresis analysis of FGF21 products after ribonuclease R treatment.

[0019] Figure 3 The expression of the FGF21 variant transfected into the supernatant of 293T cells with the corresponding circular RNA is shown.

[0020] Figure 4 The addition of FGF21 variants expressed by circular RNA-FGF21-003 (SEQ ID NO: 17) or circular RNA-FGF21-008 (SEQ ID NO: 22) to the β-Klotho overexpressing 293T cell line (A) and 3T3-L1 cell line (B) resulted in rapid phosphorylation of its downstream target ERK1 / 2.

[0021] Figure 5 The study showed that the FGF-21 variant encoded by the circular RNA stimulated the activation of the human β-Klotho receptor, thereby increasing luciferase activity.

[0022] Figure 6 (A and B) show that the in vitro activity of the FGF-21 variants encoded by the circular RNA (SEQ ID NO: 22, 36, 42, 48, 54, 60 and 61) is superior to that of the recombinant human FGF21 protein (SEQ ID NO: 74).

[0023] Figure 7 The study showed that treatment with FGF-21 variants encoded by circular RNA (SEQ ID NO: 22, 36, 42 and 54) improved oral glucose tolerance in BKS-db wild-type mice.

[0024] Figure 8 The results showed that the expression of circular RNA-FGF21-008 in the serum of representative mice increased 6 hours after administration and remained detectable until day 20 after administration.

[0025] Figure 9 The study showed that circular RNA-FGF21-003 and circular RNA-FGF21-008 reduced body weight gain in BKS-db / db mice.

[0026] Figure 10 The results showed that circular RNA-FGF21-003 and circular RNA-FGF21-008 reduced serum total cholesterol (CHOL) levels in BKS-db mice.

[0027] Figure 11 The results showed that circular RNA-FGF21-003 and circular RNA-FGF21-008 reduced serum LDL-C levels in BKS-db / db mice.

[0028] Figure 12 (A and B) show histological analysis of liver biopsies from Western diet-induced BKS-db / db mice treated with solvent, smegglutinin, and circular RNA-FGF21-008. Circular RNA-FGF21-008 significantly improved hepatic steatosis in Western diet-induced BKS-db / db mice compared to smegglutinin.

[0029] Figure 13 The results showed that, compared with rhFGF21 protein, circular RNA-FGF21-008 significantly reduced the body weight of Western diet-induced BKS-db / db mice.

[0030] Figure 14The study showed that circular RNA-FGF21-008 reduced CHOL levels in Western diet-induced BKS-db / db mice.

[0031] Figure 15 The circular RNA-FGF21-008 was shown to reduce LDL-C levels in Western diet-induced BKS-db / db mice.

[0032] Figure 16 Subcutaneous injection of circular RNA-FGF21-008 showed that it reduced the body weight of DIO mice.

[0033] Figure 17 The results showed that in DIO mice, subcutaneous injection of circular RNA-FGF21-008 resulted in a significantly greater reduction in body fat than semaglutide.

[0034] Figure 18 The results showed that smegglutinin significantly reduced lean body mass in DIO mice, while circular RNA-FGF21-008 maintained lean body mass.

[0035] Figure 19 Subcutaneous administration of circular RNA-FGF21-008 showed that it reduced CHOL levels in DIO mice.

[0036] Figure 20 The results showed that after a 5-hour fast, subcutaneous injection of circular RNA-FGF21-008 had a significantly better hypoglycemic effect than smegglutinin.

[0037] Figure 21 (A and B) show that intravenous or subcutaneous injection of circular RNA-FGF21-008 increased c-fos activity in regions of the mouse brain associated with food intake and appetite. Detailed Implementation

[0038] Unless otherwise defined below, all technical and scientific terms used in this application have the meanings commonly understood by those skilled in the art. The term "technique" as used herein is intended to refer to techniques generally understood in the art, including modifications or equivalent substitutions that are obvious to those skilled in the art. While those skilled in the art should be able to readily understand the following terms, the definitions are provided below for better explanation of this application.

[0039] As used in this application, the terms “comprising,” “including,” “having,” “containing,” or “including,” and variations thereof, are inclusive or open-ended and do not exclude other elements or method steps not listed.

[0040] As used herein, the terms “implementation,” “disclosed in this application,” or “disclosure” are not intended to be limiting but are generally applicable to any embodiment as defined in the claims or described in this application. These terms may be used interchangeably in this application.

[0041] As used in this application, the terms "treat / treating / treatment" refer to the elimination, reduction, or improvement of a disease or condition, and / or related symptoms. While not absolutely prohibited, treating a disease or condition does not require the elimination of that disease, condition, or related symptoms. The term "treatment" and its synonyms refer to the administration of a therapeutically effective amount of the circular RNA or composition disclosed in this application to a subject who needs to receive the treatment. Treatment can be directed at symptoms, for example, suppressing symptoms. Symptoms can be affected in the short term, treated in the medium term, or treated long-term, such as in a maintenance therapy setting.

[0042] Throughout the specification, the term "a / an" refers to one or more of the same entity; for example, "(a) nucleotide" is understood to mean one or more polynucleotides. Therefore, the terms "a / an," "one or more," and "at least one" are used interchangeably in this application.

[0043] As used herein, the term "variant" refers to a peptide that differs from the described peptide due to amino acid substitutions, deletions, insertions, and / or modifications. Variants can be generated using mutagenesis techniques known in the art.

[0044] The term "composition" or "pharmaceutical composition" refers to a composition comprising the circular RNA provided in this application, together with, for example, a pharmaceutically acceptable carrier, excipient, or diluent, for administration to a subject in need of treatment.

[0045] The term "pharmaceutically acceptable" means that, within reasonable medical judgment, exposure to human and animal tissues is appropriate without causing excessive toxicity or other complications commensurate with a reasonable benefit / risk ratio.

[0046] "Effective dose" refers to the amount of the circular RNA provided in this application, which, when administered to a subject as a single dose or a portion of a series of doses, is effective in treating the condition. For example, in the case of obesity, the dose of the drug is effective if taking the drug results in weight loss or maintenance of weight (e.g., prevention of weight gain), reduction of body fat, prevention or regulation of hypoglycemia, prevention or regulation of hyperglycemia, promotion of insulin synthesis, or reduction of food intake, or if so, the dose may be a fixed dose for all subjects to be treated; or it may vary depending on the subject's weight, health status and physical condition, desired weight loss or weight maintenance, the formulation of the circular RNA or composition disclosed in this application, a professional assessment of the medical condition, and other relevant factors.

[0047] The term "subject" refers to any subject who requires treatment with the circular RNA or composition provided in this application, particularly a mammalian subject. Mammal subjects include, but are not limited to, humans, dogs, cats, guinea pigs, rabbits, rats, mice, horses, cattle, bears, cattle, apes, monkeys, orangutans, chimpanzees, etc. In one embodiment, the subject is a human subject.

[0048] The term "YTE mutation" refers to a mutation at the CH2-CH3 interface that forms a contact residue for FcRn binding. According to the EU index of Kabat numbers, the YTE mutation for IgG1 Fc, IgG2 Fc, IgG3 Fc, or IgG4 Fc is M252Y / S254T / T256E.

[0049] Fibroblast growth factor 21 (FGF21) is a peptide hormone secreted by the liver, encoded by the FGF21 gene in humans. The wild-type amino acid sequence of FGF21 is shown in SEQ ID NO: 1.

[0050] The term "circular RNA" can refer to both modified and unmodified circular RNA. RNA modification is known in the art, and those skilled in the art can choose appropriate modifications if necessary.

[0051] The terms “metabolic dysfunction-associated steatohepatitis (MASH)” and “non-alcoholic steatohepatitis (NASH)” are used interchangeably. Those skilled in the art will know that MASH is also known as NASH.

[0052] The terms “metabolic dysfunction-associated fatty liver disease (MAFLD)” and “non-alcoholic fatty liver disease (NAFLD)” are used interchangeably. Those skilled in the art will know that MAFLD is also known as NAFLD.

[0053] As used in this application, the term “about” includes ±10% of the stated number. Therefore, “about 10” refers to 9-11.

[0054] As used in this article, the term "ABD" stands for albumin-binding domain. Albumin-binding domain (ABD) proteins are typically derived from bacteria and genetically engineered to bind with high affinity to serum albumin in vivo.

[0055] As used herein, the term "anti-albumin VHH" refers to the heavy chain variable domain of a heavy chain antibody against human serum albumin. Anti-albumin VHH can selectively bind to serum albumin from multiple species with high affinity.

[0056] As used in this article, the term “GLP-1” refers to glucagon-like peptide-1.

[0057] Circular RNA As used herein, the terms "circRNA," "circular polynucleotide," or "circular RNA" are used interchangeably and refer to polynucleotides that form a circular structure through covalent or non-covalent bonds. When circular RNA is mentioned, those skilled in the art will understand that the polynucleotide in RNA refers to a polynucleotide.

[0058] In one aspect, this application provides a circular RNA comprising a regulatory element and an expression element, said expression element comprising a polynucleotide encoding FGF21 or a variant thereof.

[0059] In some embodiments, the circular RNA comprises, for example, SEQ ID NO: 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 75, 76, 77, 78, 79, 80, 81, 82, 8 The nucleotide sequences shown in 3, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, or 127.

[0060] In some embodiments, the polynucleotide encoding FGF21 or a variant thereof has at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 98%, at least 99%, or 100% identity with the sequence shown in SEQ ID NO: 15, 16, 17, 18, 19, 20, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, or 113, preferably encoding FGF21 or a variant thereof, as shown in SEQ ID NO: SEQ ID NO: 15, 16, 17, 18, 19, 20, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, or 113. In a more preferred embodiment, the polynucleotide sequence encoding FGF21 or a variant thereof is as shown in SEQ ID NO: 17, 76, 89, 95, 101, 107, or 113.

[0061] In some embodiments, the amino acid sequence of FGF21 or a variant thereof is as shown in SEQ ID NO: 1, 2, 3, 4 or 74. In some embodiments, the amino acid sequence of FGF21 or a variant thereof contains one or more of the following mutations relative to wild-type FGF21 (SEQ ID NO: 1): L126R, L174P, P199G, A208E, aa1-28 deletion, and aa29-32 deletion.

[0062] In some embodiments, the expression element comprises a polynucleotide encoding Fc. In some embodiments, Fc contains one or more mutations. In some embodiments, Fc contains a YTE mutation. In some embodiments, Fc is selected from IgG1 Fc, IgG2 Fc, IgG3 Fc, and IgG4 Fc. In some preferred embodiments, Fc is IgG1 Fc or IgG4 Fc. In some embodiments, the amino acid sequence of Fc is as shown in SEQ ID NO: 65, 66, or 67.

[0063] In some embodiments, the expression element comprises a polynucleotide encoding an ABD (albumin-binding domain). In some embodiments, the amino acid sequence of the ABD has at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 98%, at least 99%, or 100% identity with the sequence shown in SEQ ID NO: 68. In some embodiments, the amino acid sequence of the ABD is as shown in SEQ ID NO: 68.

[0064] In some embodiments, the expression element comprises a polynucleotide encoding anti-albumin VHH (the heavy chain variable domain of a heavy chain antibody). In some embodiments, the amino acid sequence of VHH has at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 98%, at least 99%, or 100% identity with the sequence shown in SEQ ID NO: 69. In some embodiments, the amino acid sequence of anti-albumin VHH is as shown in SEQ ID NO: 69.

[0065] In some embodiments, the expression element comprises a polynucleotide encoding a linker between Fc and FGF21 or a variant thereof. In some embodiments, the expression element comprises a polynucleotide encoding a linker between ABD and FGF21 or a variant thereof. In some embodiments, the expression element comprises a polynucleotide encoding a linker between antialbumin VHH and FGF21 or a variant thereof. In some embodiments, the amino acid sequence of the linker is as shown in SEQ ID NO: 70, 71, 72, or 73.

[0066] In some embodiments, the circular RNA or expression element comprises, from 5' to 3', a polynucleotide encoding Fc, a polynucleotide encoding a linker, and a polynucleotide encoding FGF21 or a variant thereof. In some embodiments, the circular RNA or expression element comprises, from 5' to 3', a polynucleotide encoding ABD, a polynucleotide encoding a linker, and a polynucleotide encoding FGF21 or a variant thereof. In some embodiments, the circular RNA or expression element comprises, from 5' to 3', a polynucleotide encoding anti-albumin VHH, a polynucleotide encoding a linker, and a polynucleotide encoding FGF21 or a variant thereof.

[0067] In some embodiments, the circular RNA or expression element comprises a polynucleotide encoding GLP-1 or a variant thereof. In some embodiments, the circular RNA or expression element comprises, from 5' to 3', the following polynucleotides in sequence: a polynucleotide encoding GLP-1 or a variant thereof, a polynucleotide encoding a linker, a polynucleotide encoding Fc, a polynucleotide encoding a linker, and a polynucleotide encoding FGF21 or a variant thereof. In some embodiments, the circular RNA or expression element comprises, from 5' to 3', the following polynucleotides in sequence: a polynucleotide encoding GLP-1 or a variant thereof, a polynucleotide encoding a linker, a polynucleotide encoding ABD, a polynucleotide encoding a linker, and a polynucleotide encoding FGF21 or a variant thereof. In some embodiments, the circular RNA or expression element comprises, from 5' to 3', the following polynucleotides in sequence: a polynucleotide encoding GLP-1 or a variant thereof, a polynucleotide encoding a linker, a polynucleotide encoding anti-albumin VHH, a polynucleotide encoding a linker, and a polynucleotide encoding FGF21 or a variant thereof.

[0068] In some embodiments, the circular RNA comprises any of the polynucleotides shown in SEQ ID NOs: 21-61.

[0069] In some embodiments, the expression element comprises a sequence having at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 98%, at least 99%, or 100% identity with the sequence shown in any one of SEQ ID NOs: 21-61. Preferably, the expression element comprises a sequence shown in any one of SEQ ID NOs: 21-61. More preferably, the expression element comprises a sequence shown in SEQ ID NOs: 22, 36, 42, 48, 54, 60, and 61. Most preferably, the expression element comprises a sequence shown in SEQ ID NO: 22.

[0070] In some embodiments, the amino acid sequence encoded by the expression element of the circular RNA has at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 98%, at least 99%, or 100% identity with the sequence shown in any one of SEQ ID NOs: 5-13. Preferably, the amino acid sequence encoded by the expression element of the circular RNA comprises the sequence shown in any one of SEQ ID NOs: 5-13.

[0071] Signal peptides can guide target proteins (e.g., GLP-1 or its variants, and / or FGF21 or its variants) encoded by circular RNAs disclosed in this application into the secretory pathway of host cells. The polynucleotide encoding the signal peptide is linked to the RNA sequence encoding the target protein (e.g., GLP-1 or its variants, and / or FGF21 or its variants) disclosed in this application within the correct reading frame. The polynucleotide encoding the signal peptide is typically located at the 5' end of the RNA sequence encoding the target protein (e.g., GLP-1 or its variants, and / or FGF21 or its variants), or at the 5' end of the RNA sequence encoding Fc, ABD, or antialbumin VHH disclosed in this application.

[0072] In some embodiments, the circular RNA or expression element comprises a polynucleotide encoding a signal peptide. In some embodiments, the circular RNA or expression element comprises, from 5' to 3', the following polynucleotides in sequence: a polynucleotide encoding a signal peptide, a polynucleotide encoding Fc, a polynucleotide encoding a linker, and a polynucleotide encoding FGF21 or a variant thereof. In some embodiments, the circular RNA or expression element comprises, from 5' to 3', the following polynucleotides in sequence: a polynucleotide encoding a signal peptide, a polynucleotide encoding ABD, a polynucleotide encoding a linker, and a polynucleotide encoding FGF21 or a variant thereof. In some embodiments, the circular RNA or expression element comprises, from 5' to 3', the following polynucleotides in sequence: a polynucleotide encoding a signal peptide, a polynucleotide encoding anti-albumin VHH, a polynucleotide encoding a linker, and a polynucleotide encoding FGF21 or a variant thereof.

[0073] In some embodiments, the circular RNA or expression element comprises, from 5' to 3', the following polynucleotides in sequence: a polynucleotide encoding a signal peptide, a polynucleotide encoding GLP-1 or a variant thereof, a polynucleotide encoding a linker, a polynucleotide encoding Fc, a polynucleotide encoding a linker, and a polynucleotide encoding FGF21 or a variant thereof. In some embodiments, the circular RNA or expression element comprises, from 5' to 3', the following polynucleotides in sequence: a polynucleotide encoding a signal peptide, a polynucleotide encoding GLP-1 or a variant thereof, a polynucleotide encoding a linker, a polynucleotide encoding ABD, a polynucleotide encoding a linker, and a polynucleotide encoding FGF21 or a variant thereof. In some embodiments, the circular RNA or expression element comprises, from 5' to 3', the following polynucleotides in sequence: a polynucleotide encoding a signal peptide, a polynucleotide encoding GLP-1 or a variant thereof, a polynucleotide encoding a linker, a polynucleotide encoding anti-albumin VHH, a polynucleotide encoding a linker, and a polynucleotide encoding FGF21 or a variant thereof.

[0074] In some embodiments, the amino acid sequence of the signal peptide is shown as SEQ ID NO: 62, 63 or 64.

[0075] In some embodiments, the amino acid sequence of GLP-1 or a variant thereof is shown in SEQ ID NO: 114.

[0076] In some embodiments, the polynucleotide sequence encoding GLP-1 or a variant thereof is shown as SEQ ID NO: 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126 or 127.

[0077] In some embodiments, the polynucleotide encoding GLP-1 or a variant thereof has at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 98%, at least 99%, or 100% identity with the sequence shown in SEQ ID NO: 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, or 127.

[0078] The circular RNA disclosed in this application has the beneficial effects of reducing body weight without reducing lean muscle mass and / or reducing fat without reducing lean muscle mass.

[0079] The circular RNA disclosed in this application has the effects of reducing body weight, lowering total cholesterol levels, reducing fasting blood glucose concentration, inhibiting food intake, reducing hepatic steatosis and / or reducing fat without changing lean muscle mass.

[0080] The circular RNA disclosed in this application can be prepared using methods in the prior art. In a preferred embodiment, the circular RNA is prepared using the method described in the embodiments of this application.

[0081] Control element A regulatory element may include a sequence located adjacent to an expression element that encodes an expressed product. The regulatory element may be operatively linked to a neighboring sequence. The regulatory element may increase the amount of expressed product compared to the amount expressed in the absence of a regulatory element. Furthermore, a single regulatory element may increase the amount of product expressed by multiple tandem expression sequences. Therefore, a single regulatory element may enhance the expression of one or more expression sequences.

[0082] In some embodiments, the regulatory element includes an internal ribosome entry site (IRES) or a fragment thereof.

[0083] A suitable IRES element contained in a cyclic polynucleotide comprises an RNA sequence capable of binding to a eukaryotic ribosome. In some embodiments, the IRES element is at least about 5 nt, at least about 8 nt, at least about 9 nt, at least about 10 nt, at least about 15 nt, at least about 20 nt, at least about 25 nt, at least about 30 nt, at least about 40 nt, at least about 50 nt, at least about 100 nt, at least about 200 nt, at least about 250 nt, at least about 350 nt, or at least about 500 nt.

[0084] In some embodiments, the IRES is selected from: Coxsackievirus B3 (CVB3) IRES, Enterovirus 71 (EV71) IRES, Encephalomyocarditis Virus (EMCV) IRES, Picornavir Virus (PV) IRES, Hepatitis C Virus (HCV) IRES, Adenovirus (AdV) IRES, Human Papillomavirus 31 (HPV31) IRES, Human Herpesvirus (HHV) IRES, Raul's Sarcoma Virus (RSV) IRES, Classical Swine Fever Virus (CSFV) IRES, FGF9 IRES, SLC7A1 IRES, and RUNX1 IRES. In a preferred embodiment, the IRES is a CVB3 IRES (SEQ ID NO: 14).

[0085] Administering circular RNA to subjects In one aspect, this application provides a composition comprising the circular RNA as described in any of the preceding claims, wherein the composition further comprises a pharmaceutically acceptable excipient.

[0086] In some embodiments, the composition comprises nanoparticles, such as lipid nanoparticles.

[0087] In some embodiments, the circular RNA is administered either as naked circular RNA or as a pharmaceutical composition comprising a pharmaceutically acceptable excipient. In non-limiting examples, the pharmaceutically acceptable excipient is polyethyleneimine (PEI) or lipid nanoparticles (LNP). Other examples of liposomes that can be used to administer the circular RNA or composition include: protamine, cationic nanoemulsions, modified dendritic macromolecular nanoparticles, protamine liposomes, cationic polymers, cationic polymer liposomes, polysaccharide particles, cationic lipid nanoparticles, cationic lipid-cholesterol nanoparticles, cationic lipid-cholesterol PEG nanoparticles, cationic lipid transfection reagents sold under the trademark LIPOFECTAMINE, non-liposome transfection reagents sold under the trademark FUGENE, or any combination thereof, all of which may be used as pharmaceutically acceptable excipients.

[0088] In some embodiments, the pharmaceutical composition may optionally contain one or more other active substances, such as substances with therapeutic and / or preventative activities.

[0089] disease In one aspect, this application provides a method for treating a subject's disease, comprising administering to the subject a therapeutically effective amount of the circular RNA or composition disclosed in this application.

[0090] In one aspect, this application provides the use of the circular RNA or composition disclosed herein in the preparation of a medicament for treating a subject’s disease.

[0091] In one aspect, this application provides the circular RNA or composition disclosed herein for use in treating a subject's disease.

[0092] In some implementations, the disease is selected from: diabetes, type 1 diabetes, type 2 diabetes, malnutrition-associated diabetes, diabetic hyperosmolar hyperglycemic state, hypoglycemia in the context of diabetes, diabetic ketoacidosis, diabetic coma, uncontrolled and unstable diabetes.

[0093] In some implementations, the disease is selected from: hyperlipoproteinemia, hypercholesterolemia, hypertriglyceridemia, mixed hyperlipoproteinemia, hyperalpha lipoproteinemia, hypolipoproteinemia, hypoalpha lipoproteinemia, and hypobeta lipoproteinemia.

[0094] In some implementations, the disease is selected from: obesity or specific nutritional excesses and malnutrition.

[0095] In some implementations, the disease is defined as reducing the risk of cardiovascular events.

[0096] In some implementation schemes, the subjects are adults with type 2 diabetes and a history of cardiovascular disease.

[0097] In some implementations, the disease is selected from: metabolic dysfunction-associated fatty liver disease (MAFLD) or steatohepatitis (MASH).

[0098] In some implementations, the disease is hepatic steatosis.

[0099] While the description of the pharmaceutical compositions provided in this application is primarily directed toward pharmaceutical compositions suitable for administration to humans, those skilled in the art will understand that such compositions are generally also suitable for administration to any other animal, such as non-human animals, for example, non-human mammals.

[0100] Example To make the objectives and technical solutions of this application clearer, the application will be further described below with reference to specific embodiments. It should be understood that these embodiments are not intended to limit the scope of this application. Furthermore, specific experimental methods not mentioned in the following embodiments are all performed according to conventional experimental methods.

[0101] Example 1: Vector Construction [Amended on April 3, 2025, pursuant to Rule 91 of the PCT Rules] The coding sequences for different FGF21 variants have been internally optimized. The optimized cDNA sequences (corresponding to SEQ ID NOs: 15-61) were synthesized by Azenta Life Sciences. Typically, the synthesized fragments are cloned into the pUC57 plasmid via the Spe I / EcoRI site, as... Figure 1 As shown. The circular RNA plasmid was linearized by adding BspQ I (Vazyme, DD4302-PC-02) and incubating at 50°C for 2 hours. In this study, the heavy chain variable domain (VHH) (SEQ ID NO: 69) of a heavy chain antibody containing / without modified human IgG Fc variants (SEQ ID NOs: 65-67), an albumin-binding domain (SEQ ID NO: 68), and anti-human serum albumin (HSA) was covalently linked to an FGF21 analog.

[0102] Example 2: Production and purification of circular RNA Circular RNA precursors were synthesized from a linearized circular RNA plasmid template via in vitro transcription (IVT) using T7 RNA polymerase (Vazyme, DD4101-PC-03). Following IVT, the RNA product was treated with DNase I (Vazyme, DD4104-PC-01) at 37°C for 15 min to remove the DNA template, and then purified using an RNA purification kit (NEB, T2040L). The purified circular RNA precursors (coding regions SEQ ID NO: 17, 22, 28, 29) were ligated and circularized using T4 RNA ligase II (Kactus, TRL-BE103-C1) at 25°C for 2 h. After ligation and purification, the circular RNA product was treated with ribonuclease R (Novoprotein, GMP-E224-M001) at 37°C for 30 min to remove linear RNA. Figure 2 ).

[0103] Purified circular RNA was separated by denaturing a urea-polyacrylamide gel electrophoresis gel (Beyotime, R0218S). Briefly, 200 ng of RNA was mixed with an equal volume of 2× RNA loading staining buffer (NEB, B0363S), heated at 70°C for 10 minutes, and then immediately placed on ice. In the denaturing gel electrophoresis, ssRNA gradient standards (NEB, N0362S) were used as the RNA molecular weight standard. After electrophoresis (150V, 1 hour), the gel was stained with YeaRed nucleic acid gel dye (Yeason, 10203ES76), and the RNA bands were visualized using a ChemiDoc MP imaging system (BioRad).

[0104] Example 3: Production and purification of linear RNA Linear RNA (SEQ ID NO: 35) was synthesized from a plasmid template via in vitro transcription (IVT) using T7 RNA polymerase (Vazyme, DD4101-PC-03). After IVT, the RNA product was treated with DNase I (Vazyme, DD4104-PC-01) at 37°C for 15 minutes to remove the DNA template, and then purified using an RNA purification kit (NEB, T2040L).

[0105] Example 4: Transfection of mammalian cells and expression of circular or linear RNA 293T cells were cultured in a 37°C, 5% CO2 incubator, and supplemented with 10% fetal bovine serum (Corning, 35-081-CV) and 1% penicillin / streptomycin (Invitrogen, 15140148) in basal DMEM medium (Gibco, C11965500BT). Cells were passaged every 2-3 days. Before transfection, 3 × 10⁶ cells were... 5 Cells / well were seeded into 24-well plates. Next, 500 ng of ribonuclease R-treated circular RNA was transfected into the cells using Lipofectamine RNAiMAX (Invitrogen, 13778030).

[0106] Cell supernatant was collected 16 hours post-transfection and then subjected to Western blotting to analyze protein expression. In short, supernatants of FGF21 variants (coding sequences SEQ ID NO: 22, 28, 29, 35, 36, 42, 48, 54, 60, and 61) were separated in a 4–12% gradient SDS-PAGE (120V, 120 min), and the proteins were transferred to a PVDF membrane via semi-dry transfer (2.5V, 30 min). After blocking, the membrane was immunoblotted using anti-human IgG-Fc (Sino Biological, SSA001) and anti-FGF21 (Proteintech, 26272-1-AP). The protein can be secreted into the cell supernatant and can be detected by Western blotting. Figure 3 Furthermore, the expression levels of target proteins translated from circular RNA were higher than those translated from linear RNA.

[0107] Example 5: Circular RNA encoding an FGF21 variant stimulates ERK phosphorylation 293T and 3T3-L1 cells were cultured at 37°C and 5% CO2 in basal DMEM medium (Gibco, C11965500BT) supplemented with 10% fetal bovine serum (Corning, 35-081-CV) and 1% penicillin / streptomycin (Invitrogen, 15140148). Prior to stimulation, the expression plasmid encoding human β-Klotho (KLB) was transiently transfected into 293T and 3T3-L1 cells for 18 hours. After starvation for 3 hours, 293T and 3T3-L1 cells were treated with 300 μL of cell supernatant containing the FGF21 variant (as described in Example 4 above) for 15 min, 30 min, 1 h, 2 h, 4 h, 6 h, and 24 h. At the corresponding time points, cells were collected and lysed in buffer containing inhibitors of phosphatase and protease (Beyotime, P1045). Immunoblot analysis of cell lysates was performed using anti-phosphorylated ERK (CST, 4370T) and anti-ERK (CST, 9107S). ERK phosphorylation was induced within 15 minutes of stimulation of 293T and 3T3-L1 cells with a circular RNA-encoded FGF21 variant. Figure 4 ).

[0108] Example 6: In vitro β-Klotho luciferase reporter gene detection Luciferase reporter gene assays were performed in 293T cells stably transfected with human β-Klotho and a reporter gene construct containing 5×UAS luciferase and a GAL4 DNA-binding domain fused to Elk1. In this system, luciferase expression was regulated by endogenous phosphorylated ERK signaling. 293T-Klotho-luciferase cells were cultured at 37°C and 5% CO2 in basal DMEM medium (Gibco, C11965500BT) supplemented with 10% fetal bovine serum (Corning, 35-081-CV), 1% penicillin / streptomycin (Invitrogen, 15140148), 5 μg / mL blastomycin (Gibco, A1113903), and 2 μg / mL puromycin (Gibco, A1113803). After a 3-hour starvation period, 293T-Klotho-luciferase cells were treated with 300 μL of cell supernatant containing the FGF21 variant, as described in Example 4 above. Cells were collected 24 hours after treatment. Firefly luciferase activity was monitored using a Steady-Glo luciferase detection system (Promega, E2520). Luminescence intensity was measured using a Spark multi-well microplate reader (TECAN). The circular RNA-encoded FGF-21 variant activated the ERK-driven luciferase reporter gene in stable cell lines. Figure 5 ).

[0109] Example 7: In vitro potency of FGF21 variants encoded by circular RNA and recombinant FGF21 protein Recombinant human FGF21 protein (rhFGF21 protein, SEQ ID NO: 74) was expressed and purified in HEK293 cells (Biointron Biological Inc.). Furthermore, circular RNAs-FGF21-008,-FGF21-022,-FGF21-028,-FGF21-034,-FGF21-040,-FGF21-046, and-FGF21-047 were transfected into 293T cells. The supernatant was collected 24 hours post-transfection. The concentration of FGF21 protein in the supernatant was determined by ELISA. In short, the supernatant was diluted 1:50 in PBS and then added to microplates pre-coated with anti-FGF21 antibody (Beyotime, PF313). The colorimetric assay using tetramethylbenzidine (TMB) substrate was read at an optical density of 450 nm. A standard curve for the ELISA was plotted using a four-parameter fitting algorithm, and the measured OD450 nm values ​​of the supernatant were correlated with the standard curve.

[0110] As described in Example 6, the in vitro activity of the FGF21 variant and recombinant FGF21 protein was determined using a luciferase reporter gene assay. Briefly, supernatants containing either FGF21 or rhFGF21 protein were serially diluted twofold. Next, 293T-Klotho-luciferase cells, starved for 2 hours, were treated with 100 μL of supernatant containing the FGF21 variant or a dilution containing rhFGF21 protein. Firefly luciferase reporter gene expression was detected using a Steady-Glo luciferase detection system (Promega, E2520). Luminescence intensity was measured using a Spark multi-well microplate reader (TECAN). EC50 values ​​were analyzed using a four-parameter model. Figure 6 As shown, the in vitro activity of circular RNA-FGF21-008, circular RNA-FGF21-022, circular RNA-FGF21-028, circular RNA-FGF21-034, or circular RNA-FGF21-040 is stronger than that of rhFGF21.

[0111] Example 8: Oral glucose tolerance test (OGTT) in BKS-db wild-type mice Male BKS-db wild-type mice (8–9 weeks old) were housed in an SPF-grade environment (Jiangsu Jicui Pharmaceutical Biotechnology Co., Ltd.). Mice were divided into 5 groups based on blood glucose levels and body weight. After fasting for 5–6 hours, animals were administered 0.35 nmol / kg of circular RNA-FGF21-008, circular RNA-FGF21-022, circular RNA-FGF21-028, circular RNA-FGF21-040, or a solvent via intravenous injection. All animals were orally administered 1 g / kg of glucose solution 4 hours after administration. Blood glucose levels were measured at 15, 30, 60, and 120 minutes post-experiment. Circular RNA variants encoding FGF21 with different half-life prolongation strategies were found to reduce blood glucose levels (…). Figure 7 ).

[0112] Example 9: Pharmacokinetics of C57BL / 6 mice Six male C57BL / 6 mice were administered 0.7 nmol / kg of circular RNA-FGF21-008 via a single intravenous injection. Blood samples were collected before administration (0 h) and at 2 h, 6 h, 24 h, 72 h, 120 h, 168 h, 240 h, 336 h, 408 h, and 480 h after administration. Serum concentrations of the circular RNA encoding the FGF21 variant were determined using an ELISA method. Briefly, serum was diluted 1:50 with PBS and then added to microplates pre-coated with anti-FGF21 antibody (Beyotime). Colorimetric readings using a tetramethylbenzidine (TMB) substrate were performed at 450 nm.

[0113] A standard curve for ELISA was plotted using a four-parameter fitting algorithm, and the OD450nm value of serum was measured. Figure 8 The values ​​were correlated with a standard curve. Plasma concentrations obtained by ELISA were used to determine the pharmacokinetic parameters they represented. Serum clearance and steady-state volume of distribution were calculated using non-compartmental analysis. The mean elimination half-life was approximately 2.3 days. The half-lives of FGF21-Fc encoded by linear mRNA or rhFGF21 have been reported to be 8.9 hours and 7.1 hours, respectively (Bartesaghi, Stefano et al. Molecular therapy. Nucleic acids. 2022); however, the half-life observed after administration of the circular RNA-FGF21-008 was more than 6-fold longer than that of linear mRNA or rhFGF21.

[0114] Example 10: Therapeutic effects of Western diet-induced BKS-db / db mice SPF-grade male BKS-db / db mice (8–9 weeks old) (Jiangsu Jicui Pharmaceutical Biotechnology Co., Ltd.) were housed under 12-hour / 12-hour light / dark cycles with free access to food and water. Mice were acclimatized to the environment for one week before the study began. Based on blood glucose levels and body weight, the mice were randomly divided into four groups. After grouping, diabetic BKS-db / db mice were fed a Western diet during the experiment. Mice were injected with specified doses of test drugs (semaglutide, 10 nmol / kg subcutaneously, every three days; circular RNA-FGF21-003, 1.1 nmol / kg intravenously, once a week; circular RNA-FGF21-008, 0.7 nmol / kg intravenously, once a week) or solvents for four weeks. Body weight was recorded twice weekly. The circular RNA used here, FGF21-003, is a circular RNA composed of a CVB3-IRES sequence (SEQ ID NO: 14) and the coding sequence of FGF21 variant 1 (SEQ ID NO: 17). The circular RNA used here, FGF21-008, is a circular RNA composed of a CVB3-IRES sequence (SEQ ID NO: 14) and the coding sequence of SP1-IgG1 Fc-linker 1-FGF21 variant 2 (SEQ ID NO: 22). Blood lipid levels were measured using a chemiluminescence analyzer (Hitachi). HbA1c levels were estimated using an Afinion 2 analyzer. Therefore, mice treated with these two FGF21 variants showed significant weight loss. Figure 9 A test 28 days after treatment revealed that total cholesterol ( Figure 10 ) and low-density lipoprotein cholesterol (LDL-C) Figure 11 The level decreased significantly.

[0115] Western diet-induced BKS-db / db mice exhibit features of metabolic dysfunction-associated fatty liver disease (MASLD), including hepatic steatosis, lobular hepatitis, and ballooning degeneration of the liver. Therefore, hematoxylin-eosin staining of liver sections was performed to reveal the effect of circular RNA-FGF21-008 on liver pathology. Indeed, circular RNA-FGF21-008 slightly alleviated ballooning degeneration and lobular hepatitis. Compared to smegglutinin, circular RNA-FGF21-008 significantly reduced hepatic steatosis (…). Figure 12 (See Table 1). Overall, circular RNA-FGF21-008 improved metabolic dysfunction induced by Western diet in BKS-db / db mice.

[0116] Table 1:

[0117] Example 11: Effects of circular RNA administered via different routes on Western diet-induced BKS-db / db mice Male diabetic BKS-db / db mice (8-9 weeks old) (Jiangsu Jicui Pharmaceutical Biotechnology Co., Ltd.) were used. Mice were divided into four groups based on blood glucose levels and body weight. After grouping, diabetic BKS-db / db mice were fed a Western diet during the experiment. Wild-type BKS mice continued to be fed a normal diet. Mice were treated for four weeks with 5.4 nmol / kg rhFGF21 protein (subcutaneous injection, once a week), 0.7 nmol / kg circular RNA-FGF21-008 (intravenous or subcutaneous injection, once a week), or a solvent. Body weight was recorded weekly. Blood lipid levels were measured using a chemiluminescence analyzer (Hitachi). After treatment, rhFGF21 protein and circular RNA-FGF21-008 reduced body weight (…). Figure 13 In particular, subcutaneous injection of circular RNA-FGF21-008 resulted in greater weight loss compared to rhFGF21 protein. Furthermore, compared to the solvent group, circular RNA-FGF21-008 significantly reduced total cholesterol levels. Figure 14 ) and LDL-C levels ( Figure 15 ).

[0118] Example 12: Anti-obesity effect of circular RNA-FGF21 on diet-induced obesity (DIO) mice A diet-induced obesity rodent model was established by feeding mice with a high-fat diet for more than 10 weeks (Jiangsu Jicui Pharmaceutical Biotechnology Co., Ltd.). After a one-week adaptation period, mice were randomly divided into three groups based on blood glucose levels and body weight. DIO mice were administered 10 nmol / kg semaglutide (subcutaneous injection, every three days), 0.7 nmol / kg circular RNA-FGF21-008 (subcutaneous injection, once a week), or a solvent for four weeks. Body weight was recorded twice weekly. At the end of the study, body composition was measured using an MRI system (QMR-06-060H body composition analyzer, Numai, China). Blood lipid levels were assessed using a chemistry analyzer (Hitachi). Subcutaneous injection of circular RNA-FGF21-008 reduced the body weight of DIO mice. The weight loss induced by circular RNA-FGF21-008 was comparable to that semaglutide-treated DIO mice. Figure 16 Consistent with other reports, semaglutide treatment significantly reduced fat and lean body mass. However, circular RNA-FGF21-008 significantly reduced fat mass. Figure 17 ), while maintaining lean body mass ( Figure 18 Furthermore, compared to solvents, both smegglutinin and cyclic RNA-FGF21-008 can reduce total cholesterol levels. Figure 19 Therefore, compared to smegglutinin, treatment with circular RNA-FGF21-008 showed superior weight loss, mainly due to the reduction in fat mass and the maintenance of lean body mass.

[0119] Simultaneously, blood glucose concentrations were measured after a 5-hour fast. Furthermore, an oral glucose tolerance test (OGTT) was performed 5-6 hours after fasting, with all animals orally administered a 1 g / kg glucose solution. Blood glucose levels were measured at 15, 30, 60, and 120 minutes after glucose treatment. Fasting blood glucose levels in DIO mice were... Figure 20 Decreased blood glucose levels were observed in the OGTT (Table 2), indicating that glucose homeostasis was restored in DIO mice after treatment with circular RNA-FGF21-008. In DIO mice, circular RNA-FGF21-008 produced more significant weight loss and blood glucose reduction than smegglutinin.

[0120] Table 2:

[0121] Example 12: Activation of c-fos signaling in the mouse central nervous system Male C57BL / 6J mice were housed in a specific pathogen-free facility at Youshu Life Sciences Co., Ltd. To investigate the brain regions activated by circular RNA-FGF21-008 (labeled with c-fos) and their mediating of appetite, mice were administered semaglutide subcutaneously for 4 hours (10 nmol / kg), intravenously for 6 hours (0.7 nmol / kg), or subcutaneously for 24 hours (0.7 nmol / kg). Brain tissue was collected, fixed overnight in 4% formalin at 4°C, followed by standard dehydration and paraffin embedding. Immunofluorescence staining was performed on sections to detect c-fos signal transduction in different brain regions (Servicebio). Quantitative analysis of c-fos signal intensity in each brain region was performed using Fiji software at 20x magnification.

[0122] Following administration of circular RNA-FGF21-008, increased c-fos activity was observed in the brainstem, including areas such as the organelle of the lamina terminalis (OV) and the area extremely behind the lamina (AP), and in the forebrain, including areas such as the bed nucleus of the stria terminalis (BST), the lateral preoptic area (LPO), the central amygdala (CeA), the lateral hypothalamus (LHA), the parathalamic nucleus (PSTN), and the dorsal midline thalamic group (MTN). Figure 21 The aforementioned brain regions are associated with food intake and appetite; however, c-fos signaling is not activated in the subfornix organ (SFO), parabrachial nucleus (PB), dorsal motor nucleus of the vagus nerve (DMX), and nucleus of the solitary tract (NTS). Even in the absence of the blood-brain barrier, FGF21 receptors may not be abundant in the SFO and PB regions. Furthermore, the DMX and NTS regions are protected by the blood-brain barrier, making it difficult for circular RNA-FGF21-008 to cross the barrier and activate its cascade signal transduction. Notably, the activation of c-fos activity in the BST, CeA, and AP by circular RNA-FGF21-008 differs slightly, possibly due to different routes of administration.

[0123] Since the methods, compounds, and compositions of this application have been fully described, those skilled in the art will understand that the same operations can be performed under a wide range of equivalent conditions, formulations, and other parameters without affecting the scope of the methods, compounds, and compositions or any embodiments thereof provided in this application.

[0124] All patents, patent applications and publications cited in this application are incorporated herein by reference in their entirety.

[0125] sequence list The nucleotide sequences shown in this application may represent RNA or amino acid sequences and can be converted into the WIPO standard ST.26 sequence listing.

[0126] Table 1: Serial Numbers Used in This Application

[0127] amino acid sequence of wild-type FGF21 SEQ ID NO: 1 MDSDETGFFEHSGLWVSVLAGLLLGACQAHPIPDSSPLLQFGGQVRQRYLYTDDAQQTEAHLEIREDGTVGGAADQSPESLLQLKALKPGVIQILGVKTSRFLCQRPDGALYGSLHFDPEACSFRELLLEDGYNVYQSEAHGLPLHLPGNKSPHRDPAPRGPARFLPLPGLPPALPEPPGILAPQPPDVGSSDPLSMVGPSQGRSPSYAS The amino acid sequence of FGF21 variant 1 (aa1-28; aa33-209, L126R, P199G, A208E) SEQ ID NO: 2 MDSDETGFFEHSGLWVSVLAGLLLGACQADSSPLLQFGGQVRQRYLYTDDAQQTEAHLEIREDGTVGGAADQSPESLLQLKALKPGVIQILGVKTSRFLCQRPDGALYGSLHFDPEACSFRERLLEDGYNVYQSEAHGLPLHLPGNKSPHRDPAPRGPARFLPLPGLPPALPEPPGILAPQPPDVGSSDPLSMVGGSQGRSPSYES The amino acid sequence of FGF21 variant 2 (aa29-209, L126R, P199G, A208E) SEQ ID NO: 3 HPIPDSSPLLQFGGQVRQRYLYTDDAQQTEAHLEIREDGTVGGAADQSPESLLQLKALKPGVIQILGVKTSRFLCQRPDGALYGSLHFDPEACSFRERLLEDGYNVYQSEAHGLPLHLPGNKSPHRDPAPRGPARFLPLPGLPPALPEPPGILAPQPPDVGSSDPLSMVGGSQGRSPSYES Amino acid sequence of FGF21 variant 3 (aa29 - 209, L126R, L174P, P199G) SEQ ID NO: 4 HPIPDSSPLLQFGGQVRQRYLYTDDAQQTEAHLEIREDGTVGGAADQSPESLLQLKALKPGVIQILGVKTSRFLCQRPDGALYGSLHFDPEACSFRERLLEDGYNVYQSEAHGLPLHLPGNKSPHRDPAPRGPARFLPLPGLPPAPPEPPGILAPQPPDVGSSDPLSMVGGSQGRSPSYAS Amino acid sequence of SP1 - IgG1 Fc - linker 1 - FGF21 variant 2 SEQ ID NO: 5 MGWSCIILFLVATATGVHSDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKGGGGSGGGGSGGGGSHPIPDSSPLLQFGGQVRQRYLYTDDAQQTEAHLEIREDGTVGGAADQSPESLLQLKALKPGVIQILGVKTSRFLCQRPDGALYGSLHFDPEACSFRERLLEDGYNVYQSEAHGLPLHLPGNKSPHRDPAPRGPARFLPLPGLPPALPEPPGILAPQPPDVGSSDPLSMVGGSQGRSPSYES Amino acid sequence of SP2 - IgG1 Fc - linker 4 - FGF21 variant 3 SEQ ID NO: 6 METDTLLLMVLLLWVPGSTGDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKGGGGGSGGGSGGGGSHPIPDSSPLLQFGGQVRQRYLYTDDAQQTEAHLEIREDGTVGGAADQSPESLLQLKALKPGVIQILGVKTSRFLCQRPDGALYGSLHFDPEACSFRERLLEDGYNVYQSEAHGLPLHLPGNKSPHRDPAPRGPARFLPLPGLPPAPPEPPGILAPQPPDVGSSDPLSMVGGSQGRSPSYAS Amino acid sequence of SP1 - IgG1 Fc (YTE) - linker 1 - FGF21 variant 2 SEQ ID NO: 7 MGWSCIILFLVATATGVHSDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKGGGGSGGGGSGGGGSHPIPDSSPLLQFGGQVRQRYLYTDDAQQTEAHLEIREDGTVGGAADQSPESLLQLKALKPGVIQILGVKTSRFLCQRPDGALYGSLHFDPEACSFRERLLEDGYNVYQSEAHGLPLHLPGNKSPHRDPAPRGPARFLPLPGLPPALPEPPGILAPQPPDVGSSDPLSMVGGSQGRSPSYES Amino acid sequence of SP1 - ABD - linker 2 - FGF21 variant 2 SEQ ID NO: 8 MGWSCIILFLVATATGVHSAEAKEAANAELDSYGVSDFYKRLIDKAKTVEGVEALKDAILAALPGGGGSGGGSHPIPDSSPLLQFGGQVRQRYLYTDDAQQTEAHLEIREDGTVGGAADQSPESLLQLKALKPGVIQILGVKTSRFLCQRPDGALYGSLHFDPEACSFRERLLEDGYNVYQSEAHGLPLHLPGNKSPHRDPAPRGPARFLPLPGLPPALPEPPGILAPQPPDVGSSDPLSMVGGSQGRSPSYES Amino acid sequence of SP1 - anti - albumin VHH - linker 2 - FGF21 variant 2 SEQ ID NO: 9 MGWSCIILFLVATATGVHSEVQLVESGGGLVQPGNSLRLSCAASGFTFSSFGMSWVRQAPGKGLEWVSSISGSGSDTLYADSVKGRFTISRDNAKTTLYLQMNSLRPEDTAVYYCTIGGSLSRSSQGTLVTVSSGGGGSGGGSHPIPDSSPLLQFGGQVRQRYLYTDDAQQTEAHLEIREDGTVGGAADQSPESLLQLKALKPGVIQILGVKTSRFLCQRPDGALYGSLHFDPEACSFRERLLEDGYNVYQSEAHGLPLHLPGNKSPHRDPAPRGPARFLPLPGLPPALPEPPGILAPQPPDVGSSDPLSMVGGSQGRSPSYES Amino acid sequence of SP3 - GLP-1 – linker 3 - ABD - linker 2 - FGF21 variant 2 SEQ ID NO: 10 METDTLLLWVLLLWVPGSTGHGEGTFTSDVSSYLEEQAAKEFIAWLVKGGGGGGGSAEAKEAANAELDSYGVSDFYKRLIDKAKTVEGVEALKDAILAALPGGGGSGGGSHPIPDSSPLLQFGGQVRQRYLYTDDAQQTEAHLEIREDGTVGGAADQSPESLLQLKALKPGVIQILGVKTSRFLCQRPDGALYGSLHFDPEACSFRERLLEDGYNVYQSEAHGLPLHLPGNKSPHRDPAPRGPARFLPLPGLPPALPEPPGILAPQPPDVGSSDPLSMVGGSQGRSPSYES Amino acid sequence of SP3 - GLP-1 - linker 2 - anti - albumin VHH - linker 2 - FGF21 variant 2 SEQ ID NO: 11 METDTLLLWVLLLWVPGSTGHGEGTFTSDVSSYLEEQAAKEFIAWLVKGGGGGGGSGGGSEVQLVESGGGLVQPGNSLRLSCAASGFTFSSFGMSWVRQAPGKGLEWVSSISGSGSDTLYADSVKGRFTISRDNAKTTLYLQMNSLRPEDTAVYYCTIGGSLSRSSQGTLVTVSSGGGGSGGGSHPIPDSSPLLQFGGQVRQRYLYTDDAQQTEAHLEIREDGTVGGAADQSPESLLQLKALKPGVIQILGVKTSRFLCQRPDGALYGSLHFDPEACSFRERLLEDGYNVYQSEAHGLPLHLPGNKSPHRDPAPRGPARFLPLPGLPPALPEPPGILAPQPPDVGSSDPLSMVGGSQGRSPSYES Amino acid sequence of SP3 - GLP - 1 - linker1 - IgG1 Fc - linker1 - FGF21 variant 2 SEQ ID NO: 12 METDTLLLWVLLLWVPGSTGHGEGTFTSDVSSYLEEQAAKEFIAWLVKGGGGGGGSGGGGSGGGGSDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKGGGGSGGGGSGGGGSHPIPDSSPLLQFGGQVRQRYLYTDDAQQTEAHLEIREDGTVGGAADQSPESLLQLKALKPGVIQILGVKTSRFLCQRPDGALYGSLHFDPEACSFRERLLEDGYNVYQSEAHGLPLHLPGNKSPHRDPAPRGPARFLPLPGLPPALPEPPGILAPQPPDVGSSDPLSMVGGSQGRSPSYES Amino Acid Sequence of SP3 - GLP-1 - Linker1 - IgG4 Fc - Linker1 - FGF21 Variant 2 SEQ ID NO: 13 METDTLLLWVLLLWVPGSTGHGEGTFTSDVSSYLEEQAAKEFIAWLVKGGGGGGGSGGGGSGGGGSAESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGGGGGSGGGGSGGGGSHPIPDSSPLLQFGGQVRQRYLYTDDAQQTEAHLEIREDGTVGGAADQSPESLLQLKALKPGVIQILGVKTSRFLCQRPDGALYGSLHFDPEACSFRERLLEDGYNVYQSEAHGLPLHLPGNKSPHRDPAPRGPARFLPLPGLPPALPEPPGILAPQPPDVGSSDPLSMVGGSQGRSPSYES CVB3-IRES SEQ ID NO: 14 1 TTAAAACAGC CTGTGGGTTG ATCCCACCCA CAGGCCCATT GGGCGCTAGC 51 ACTCTGGTAT CACGGTACCT TTGTGCGCCT GTTTTATACC CCCTCCCCCA 101 ACTGTAACTT AGAAGTAACA CACACCGATC AACAGTCAGC GTGGCACACC 151 AGCCACGTTT TGATCAAGCA CTTCTGTTAC CCCGGACTGA GTATCAATAG 201 ACTGCTCACG CGGTTGAAGG AGAAAGCGTT CGTTATCCGG CCAACTACTT 251 CGAAAACCT AGTAACACCG TGGAAGTTGC AGAGTGTTTC GCTCAGCACT 301 ACCCCAGTGT AGATCAGGTC GATGAGTCAC CGCATTCCCC ACGGGCGACC 351 GTGGCGGTGG CTGCGTTGGC GGCCTGCCCA TGTTTGGGGA AACCCAAACA 401 TGGGACGCTC TAATACAGAC ATGGTGCGAA GAGTCTATTG AGCTAGTTGG 451 TAGTCCTCCG GCCCCTGAAT GCGGCTAATC CTAACTGCGG AGCACACACC 501 CTCAAGCCAG AGGGCAGTGT GTCGTAACGG GCAACTCTGC AGCGGAACCG 551 ACTACTTTGG GTGTCCGTGT TTCATTTTAT TCCTATACTG GCTGCTTATG 601 GTGACAATTG AGAGATCGTT ACCATATAGC TATTGGATTG GCCATCCGGT 651 GACTAATAGA GCTATTATAT ATCCCTTTGT TGGGTTTATA CCACTTAGCT 701 TGAAAGAGGT TAAAACATTA CAATTCATTG TTAAGTTGAA TACAGCA FGF21 also contains 1 protein SEQ ID NO:15 1 ATGGACAGTG ATGAAACTGG ATTTGAACAT TCAGGGCTCT GGGTTTCTGT 51 TTTAGCAGGG CTGCTGCTGG GGGCCTGCCA AGCAGACAGC AGCCCCCTGC 101 TGCAGTTTGG AGCCAGTT CGCCAGCGCT ACCTCTACAC BUYGCC 151 CAGCAAACAG AGCACATTT GAAGATGGA CTGTTGGTGG 201 GGCGGCGGAC CAGTCTCCAG AAAGTTTATT ACATTAAAAA GCATTAAAAC 251 CTGGTGTTAT TCAAATATTA GGAGTAAAAA CCAGCCGCTT CCTCTGCCAG 301 AGGCCAGATG GTGCTTTATA TGGAAGTTTA CATTTTGATC CTGAGGCCTG 351 CTCCTTCAGA GAAAGATTAT TAGAAGATGG CTACAATGTT TATCAAAGTG 401 AAGCACATGG TCTTCCTCTT CATCTTCCTG GAAATAAATC ACCCCACCGG 451 GACCCGGCGC CGCGGGGCCC GGCGCGCTTC CTTCCTCTTC CTGGGCTGCC 501 GCCGGCGCTG CCGGAGCCGC CGGGCATCCT GGCGCCGCAG CCGCCGGATG 551 TTGGCAGCAG TGACCCCCTC TCCATGGTTG GTGGCAGCCA GGGCCGCAGC 601 CCCAGCTATG AAAGT FGF21 also contains 1 protein SEQ ID NO: 16 1 ATGGACAGTG ATGAAACTGG ATTTGAACAT TCAGGGCTCT GGGTTTCTGT 51 TTTAGCAGGT TTATTATTAG GAGCCTGCCA AGCAGATTCT TCTCCTTTAT 101 TACAATTTGG GROWTH CGTCAGCGCT ACCTCTACAC GROWGCC 151 CAGCAAACAG AGCACATTT GAAGATGGA CTGTTGGTGG 201 GGCGGCGGAC CAAAGTCCTG AAAGTTTATT ACATTAAAAA GCATTAAAAC 251 CTGGTGTTAT TCAAATATTA GGAGTAAAAA CCAGCCGCTT CCTCTGCCAA 301 AGACCAGATG GTGCTTTATA TGGAAGTTTA CATTTTGATC CTGAGGCCTG 351 CTCCTTCAGA GAAAGATTAT TAGAAGATGG TTATAATGTT TATCAAAGTG 401 AAGCACATGG TCTTCCTTTA CATCTTCCTG GAAATAAATC ACCCCACCGG 451 GACCCGGCGC CGCGGGGCCC GGCGCGCTTC CTTCCTCTTC CTGGGCTGCC 501 GCCGGCGCTG CCGGAGCCGC CGGGCATCCT GGCGCCGCAG CCGCCGGATG 551 TTGGCAGCAG TGACCCCCTC TCCATGGTTG GTGGCAGCCA AGGCCGCTCG 601 CCCAGCTATG AAAGT FGF21 also contains 1 protein SEQ ID NO: 17 1 ATGGACAGTG ATGAAACTGG ATTTGAACAT TCAGGGCTCT GGGTTTCTGT 51 TTTAGCAGGT TTATTATTAG GAGCCTGCCA AGCAGATTCT TCTCCTTTAT 101 TACAATTTGG TGGTCAAGTA TACAATTTGG ATTTATATAC AGATGATGCA 151 CAACAAACAG AGCACATTT GAAGATGGA CTGTTGGTGG 201 GGCGGCGGAC CAAAGTCCTG AAAGTTTATT ACATTAAAAA GCATTAAAAC 251 CTGGTGTTAT TCAAATATTA GGAGTAAAAA CCAGCCGCTT CCTCTGCCAA 301 AGACCAGATG GTGCTTTATA TGGAAGTTTA CATTTTGATC CTGAGGCCTG 351 CTCCTTCAGA GAAAGATTAT TAGAAGATGG TTATAATGTT TATCAAAGTG 401 AAGCACATGG TCTTCCTTTA CATTTACCTG GAAATAAATC CCCGCACCGG 451 GACCCGGCGC CGCGGGGCCC GGCGCGCCTTC CTCCCGCTGC CGGGCTCCC 501 GCCGGCGCTG CCGGAGCCGC CGGGCATCCT GGCGCCGCAG CCGCCGGATG 551 TTGGCAGCAG TGACCCCCTC TCCATGGTTG GTGGCAGCCA AGGCCGCTCG 601 CCCAGCTATG AAAGT FGF21 also contains 1 protein SEQ ID NO: 18 1 ATGGACAGTG ATGAAACTGG ATTTGAGCAC AGTGGCCTCT GGGTTTCTGT 51 GCTGGCGGGC CTGCTGCTGG GGGCCTGCCA GGCTGACAGC AGCCCCCTGC 101 TGCAGTTTGG TGGCCAGGTG CGGCAGCGCT ACCTCTACAC BUYGCC 151 CAGCAGACGG AGGCCCACCT GGAGATCCGA GAAGATGGAA CTGTTGGTGG 201 GGCGGCGGAC CAGAGCCCTG AGAGCCTGCT GCAGCTGAAG GCCCTGAAGC 251 CTGGTGTCAT CCAGATCCTG GGGGTCAAGA CCAGCCGCTT CCTCTGCCAG 301 AGGCCAGATG GGGCCCTCTA TGGCAGCCTT CATTTTGATC CTGAGGCCTG 351 CTCCTTCCGG GAGCGGCTGC TGGAGGATGG CTACAATGTC TACCAGAGTG 401 AAGCTCATGG ACTTCCTCTT CATCTTCCTG GGAACAAGAG CCCCCACCGG 451 GACCCGGCGC CGCGGGGCCC GGCGCGCTTC CTGCCCCTGC CTGGGCTGCC 501 GCCGGCGCTG CCTGAGCCGC CGGGCATCCT GGCGCCCCAG CCCCCAGATG 551 TGGGCAGCAG TGACCCCCTC TCCATGGTTG GTGGCAGCCA GGGCCGCAGC 601 CCCAGCTATG AAAGT Coding sequence of FGF21 variant 1 SEQ ID NO: 19 1 ATGGACAGTG ATGAAACTGG ATTTGAGCAC TCGGGCCTCT GGGTTTCTGT 51 GCTGGCGGGC CTGCTGCTGG GGGCCTGCCA GGCTGACAGC AGCCCCCTGC 101 TGCAGTTTGG TGGCCAGGTG CGGCAGCGCT ACCTCTACAC AGATGATGCC 151 CAGCAGACAG AAGCTCATTT AGAAATAAGA GAAGATGGAA CTGTTGGTGG 201 GGCGGCGGAC CAGAGCCCTG AGAGCCTGCT GCAGCTGAAG GCGCTGAAGC 251 CAGGTGTCAT CCAGATTCTT GGTGTCAAGA CCAGCCGCTT CCTCTGCCAG 301 AGGCCAGATG GTGCTTTATA TGGCAGCCTT CATTTTGATC CTGAGGCCTG 351 CTCCTTCCGG GAGCGGCTGC TGGAGGATGG CTACAATGTC TACCAGAGTG 401 AAGCTCATGG ACTTCCTCTT CATCTTCCTG GGAACAAGAG CCCCCACCGG 451 GACCCGGCGC CGCGGGGCCC GGCGCGCTTC CTGCCCCTGC CGGGCCTCCC 501 GCCGGCGCTG CCGGAGCCGC CGGGCATCCT GGCGCCGCAG CCGCCGGATG 551 TGGGCAGCAG TGACCCCCTC TCCATGGTTG GTGGCAGCCA GGGCCGCAGC 601 CCCAGCTATG AAAGT Coding sequence of FGF21 variant 1 SEQ ID NO: 20 1 ATGGACAGTG ATGAAACTGG ATTTGAACAT TCAGGCCTCT GGGTTTCTGT 51 TTTAGCAGGG CTGCTGCTGG GGGCCTGCCA AGCAGACAGC AGCCCCCTGC 101 TGCAGTTTGG TGGCCAGGTG CGGCAGCGCT ACCTCTACAC AGATGATGCC 151 CAGCAGACAG AAGCACATTT AGAAATAAGA GAAGATGGAA CTGTTGGTGG 201 GGCGGCGGAC CAGTCTCCAG AAAGTTTATT ACAATTAAAA GCATTAAAAC 251 CTGGTGTCAT CCAGATTCTT GGAGTAAAAA CCAGCCGCTT CCTCTGCCAG 301 AGGCCAGATG GGGCCCTCTA TGGCAGCCTT CATTTTGATC CTGAGGCCTG 351 CTCCTTCAGA GAAAGATTAT TAGAAGATGG CTACAATGTT TATCAAAGTG 401 AAGCACATGG TCTTCCTCTT CATCTTCCTG GAAATAAATC TCCCCACCGG 451 GACCCGGCGC CCCGGGGCCC GGCGCGCCTTC CTGCCCCTTC CTGGGCTGCC 501 GCCGGCGCTG CCGGAGCCGC CGGGCATCCT GGCGCCCCAG CCCCCAGATG 551 TGGGCAGCAG TGACCCCCTC TCCATGGTTG GTGGCAGCCA GGGCCGCAGC 601 CCCAGCTATG AAAGT SP1 - IgG1 Fc - Protein1 - FGF21 and 2 subunit complexes SEQ ID NO: 21 1 ATGGGCTGGA GCTGCATCAT CCTCTTCCTG GTGGCCACGG CCACTGGTGT 51 TCATTCAGAT AAAACTCACA CCTGCCCGCC GTGCCCGGCG CCGGAGCTGC 101 TGGGGGGCCC CTCGGTCTTC CTCTTCCCGC CCAAGCCCA AGATACTTTA 151 ATGATCAGCC GCACGCCGGA GGTCACCTGT GTTGTTGTAG ATGTTTCTCA 201 TGAAGATCCT GAAGTAAAAT TTAACTGGTA TGTAGATGGT GTTGAAGTTC 251 ATATGCAAA AACAAAACCC CGGGAGGAGC AGTACAACAG CACCTACAGA 301 GTTGTTTCTG TTTTAACTGT TTTACATCAA GACTGGTTAA ATGGAAAAGA 351 ATATAAATGT AAAGTTTCAA ATAAAGCCCT GCCGGCGCCC ATTGAAAAAA 401 CCATCTCCAA AGCAAAAGGA CAGCCCCGGG AGCCCCAGGT CTACACGCTG 451 CCGCCCAGCA GAGATGAATT AACAAAAAAAT CAAGTTTCTC TCACCTGCCT 501 TGTAAAAGGA TTTTATCCTT CAGATATTGC TGTAGAATGG GAATCAAATG 551 GACAACCAGA AAATAATTAT AAAACCACGC CGCCGGTTTT AGATTCAGAT 601 GGCTCCTTCT TCCTCTACAG CAAATTAACT GTAGATAAA GCCGCTGGCA 651 ACAAGGAAAT GTCTTCTCCT GCAGTGTCAT GCATGAGGCG CTGCACAACC 701 ACTACACGCA GAAAAGTTTA TCTTTATCTC CTGGGAAAGG TGGTGGTGGC 751 AGTGGTGGTG GTGGCAGTGG TGGTGGTGGC AGCCACCCCA TCCCGGACAG 801 CAGCCCGCTG CTGCAGTTTG GAGGACAAGT TCGTCAGCGC TACCTCTACA 851 CAGATGATGC CCAGCAAACA GAAGCACATT TAGAAATAAG AGAAGATGGA 901 ACTGTTGGTG GGGCGGCGGA CCAAAGTCCT GAAAGTTTAT TACAATTAAA 951 AGCATTAAAA CCTGGTGTTA TTCAAATATT AGGAGTAAAA ACCAGCCGCT 1001 TCCTCTGCCA AAGACCAGAT GGTGCTTTAT ATGGAAGTTT ACATTTTGAT 1051 CCTGAGGCCT GCTCCTTCAG AGAAAGATTA TTAGAAGATG GTTATAATGT 1101 TTATCAAAGT GAAGCACATG GTCTTCCTTT ACATCTTCCT GGAAATAAAT 1151 CACCCCACCG GGACCCGGCG CCGCGGGGCC CGGCGCGCTT CCTTCCTCTT 1201 CCTGGGCTGC CGCCGGCGCT GCCGGAGCCG CCGGGCATCC TGGCGCCGCA 1251 GCCGCCGGAT GTTGGCAGCA GTGACCCCCT CTCCATGGTT GGTGGCAGCC 1301 AAGGCCGCTC GCCCAGCTAT GAAAGT Coding sequence of SP1 - IgG1 Fc - linker1 - FGF21 variant 2 SEQ ID NO: 22 1 ATGGGCTGGA GCTGCATCAT CCTCTTCCTG GTGGCCACGG CCACTGGAGT 51 TCATTCAGAT AAAACCCACA CCTGCCCGCC CTGCCCGGCG CCGGAGCTGC 101 TGGGGGGCCC CAGTGTCTTC CTCTTCCCGC CCAAGCCCAA GGACACCCTC 151 ATGATTTCAA GAACTCCAGA AGTCACCTGT GTGGTGGTGG ATGTTTCTCA 201 TGAAGATCCT GAGGTGAAGT TCAACTGGTA TGTTGATGGT GTTGAAGTTC 251 ATAATGCCAA GACCAAGCCC CGGGAGGAGC AGTACAACAG CACCTACAGA 301 GTTGTTTCTG TTTTAACTGT TCTTCATCAG GACTGGTTAA ATGGAAAAGA 351 ATATAAATGT AAAGTTTCAA ATAAAGCCCT GCCCGCGCCC ATTGAAAAAA 401 CCATCTCCAA AGCAAAAGGA CAGCCCCGGG AGCCCCAGGT GTACACGCTG 451 CCGCCCAGCA GAGATGAGCT GACCAAAAAT CAAGTCAGCC TCACCTGCCT 501 GGTGAAGGGC TTCTACCCCA GTGATATTGC TGTGGAGTGG GAGAGCAATG 551 GGCAGCCAGA AAATAATTAT AAAACCACCC CGCCGGTGCT GGACAGTGAT 601 GGCTCCTTCT TCCTCTACAG CAAATTAACT GTGGACAAGA GCCGCTGGCA 651 GCAAGGAAAT GTCTTCAGCT GCAGTGTCAT GCATGAAGCC CTGCACAACC 701 ACTACACCCA GAAAAGCCTC AGCCTCAGCC CTGGGAAAGG AGGTGGTGGC 751 AGTGGTGGTG GTGGCAGTGG TGGTGGTGGC AGCCACCCCA TCCCAGACAG 801 CAGCCCCCTG CTGCAGTTTG GTGGCCAGGT GCGGCAGCGC TACCTCTACA 851 CAGATGATGC CCAGCAGACA GAAGCACATT TAGAAATAAG AGAAGATGGA 901 ACTGTTGGTG GTGCTGCTGA CCAGAGCCCT GAGAGCCTGC TGCAGCTGAA 951 GGCCCTGAAG CCAGGTGTCA TCCAGATTCT TGGTGTGAAG ACCAGCCGCT 1001 TCCTCTGCCA GAGGCCAGAT GGGGCCCTCT ATGGCAGCCT TCATTTTGAT 1051 CCTGAGGCCT GCAGCTTCCG GGAGCGGCTG CTGGAAGATG GCTACAATGT 1101 TTATCAAAGT GAAGCACATG GCCTGCCCCT GCACCTTCCT GGAAATAAAA 1151 GCCCCCACAG AGATCCTGCC CCCCGGGGCC CTGCCCGCTT CCTGCCCCTG 1201 CCTGGGCTGC CCCCGGCCCT GCCTGAGCCG CCGGGCATCC TGGCCCCCCA 1251 GCCCCCAGAT GTGGGCAGCA GTGACCCCCT CTCCATGGTT GGTGGCAGCC 1301 AGGGCCGCAG CCCCAGCTAT GAAAGT Coding sequence of SP1 - IgG1 Fc - linker1 - FGF21 variant 2 SEQ ID NO: 23 1 ATGGGCTGGA GCTGCATCAT CCTCTTCCTG GTGGCCACGG CCACTGGTGT 51 CCACAGTGAC AAGACCCACA CCTGCCCGCC GTGCCCGGCG CCGGAGCTGC 101 TGGGGGGCCC CTCGGTCTTC CTCTTCCCGC CCAAGCCCAA GGACACGCTC 151 ATGATCAGCC GCACGCCGGA GGTCACCTGT GTTGTTGTAG ATGTTTCTCA 201 TGAAGATCCT GAAGTAAAAT TCAACTGGTA TGTAGATGGT GTAGAAGTTC 251 ATAATGCCAA GACCAAGCCC CGGGAGGAGC AGTACAACAG CACCTACAGA 301 GTTGTTTCTG TTTTAACTGT TCTTCATCAA GACTGGCTAA ATGGAAAAGA 351 ATATAAATGT AAAGTCAGCA ACAAGGCGCT GCCGGCGCCC ATTGAAAAAA 401 CCATCTCCAA GGCCAAGGGC CAGCCGCGGG AGCCCCAGGT CTACACGCTG 451 CCGCCGTCCA GAGATGAGCT CACCAAGAAC CAAGTCAGCC TCACCTGCCT 501 GGTGAAGGGC TTCTACCCCT CGGACATTGC TGTGGAGTGG GAGAGCAATG 551 GGCAGCCGGA GAACAACTAC AAGACCACGC CGCCGGTGCT GGACAGTGAT 601 GGCTCCTTCT TCCTCTACAG CAAGCTCACG GTGGACAAGA GCCGCTGGCA 651 ACAAGGAAAT GTCTTCTCCT GCAGTGTCAT GCATGAGGCG CTGCACAACC 701 ACTACACGCA GAAAAGCCTC AGCCTCTCGC CGGGGAAAGG TGGTGGTGGC 751 AGTGGTGGTG GTGGCAGTGG TGGTGGTGGC AGCCACCCCA TCCCAGACAG 801 CAGCCCGCTG CTGCAGTTTG GTGGTCAAGT TCGACAGCGC TACCTCTACA 851 CAGATGATGC CCAGCAGACA GAAGCTCATT TAGAAATAAG AGAAGATGGA 901 ACTGTTGGTG GGGCGGCGGA CCAGTCTCCA GAAAGTTTAT TACAATTAAA 951 AGCTTTAAAA CCTGGTGTCA TCCAGATTCT TGGTGTCAAG ACCAGCCGCT 1001 TCCTCTGCCA GAGGCCAGAT GGTGCTTTAT ATGGCAGCCT TCATTTTGAT 1051 CCTGAGGCCT GCTCCTTCCG AGAAAGATTA TTAGAAGATG GCTACAATGT 1101 CTACCAGAGT GAAGCTCATG GTCTTCCTCT TCATCTTCCT GGGAACAAGT 1151 CGCCGCACCG GGACCCGGCG CCGCGGGGCC CGGCGCGCTT CCTGCCGCTG 1201 CCGGGCCTCC CGCCGGCGCT GCCGGAGCCG CCGGGCATCC TGGCGCCGCA 1251 GCCGCCGGAT GTGGGCAGCA GTGACCCCCT CTCCATGGTT GGTGGCAGCC 1301 AGGGCCGCTC GCCCAGCTAT GAAAGT SP1 - IgG1 Fc - Protein1 - FGF21 and 2 subunit complexes SEQ ID NO: 24 1 ATGGGCTGGA GCTGCATCAT CCTCTTCCTG GTGGCCACGG CCACGGGTGT 51 CCACTCGGAC AAGACGCACA CGTGCCCGCC GTGCCCGGCG CCGGAGCTGC 101 TGGGGGGCCC CTCGGTCTTC CTCTTCCCGC CCAAGCCCA GGACACGCTC 151 ATGATCAGCC GCACGCCGGA GGTCACCTGT GTTGTTGTAG ATGTTTCTCA 201 TGAAGATCCT GAAGTAAAAT TCAACTGGTA TGTAGATGGT GTAGAAGTTC 251 ATATGCCAA GACCAAGCCG CGGGAGGAGC AGTACAACAG CACCTACAGA 301 GTTGTTTCTG TTTTAACTGT TCTTCATCAA GACTGGCTAA ATGGAAAAGA 351 ATATAAATGT AAAGTCAGCA ACAAGGCGCT GCCGGCGCCC ATTGAAAAAA 401 CCATCTCCAA AGCAAAAGGA CAGCCGCGGG AGCCCCAGGT CTACACGCTG 451 CCGCCGTCCA GAGATGAGCT CACCAAGAAC CAAGTCAGCC TCACCTGCCT 501 TGTAAAAGGC TTCTACCCGT CGGACATTGC TGTAGAATGG GAGAGCAATG 551 GGCAGCCGGA GAACAACTAC AAGACCACGC CGCCGGTGCT GGACAGTGAT 601 GGCTCCTTCT TCCTCTACAG CAAGCTCACG GTGGACAAGA GCCGCTGGCA 651 ACAAGGAAAT GTCTTCTCCT GCAGTGTCAT GCATGAGGCG CTGCACAACC 701 ACTACACGCA GAAGAGCCTC AGCCTCTCGC CGGGGAAAGG TGGTGGTGGC 751 AGTGGTGGTG GTGGCAGTGG TGGTGGTGGC AGCCACCCCA TCCCGGACTC 801 GTCGCCGCTG CTGCAGTTTG GTGGTCAAGT TCGACAGCGC TACCTCTACA 851 CAGATGATGC TCAACAAACA GAAGCTCATT TAGAAATAAG AGAAGATGGA 901 ACTGTTGGTG GGGCGGCGGA CCAGTCGCCG GAGAGTTTAT TACAATTAAA 951 AGCTTTAAAA CCTGGTGTCA TCCAGATTCT TGGTGTCAAG ACCAGCCGCT 1001 TCCTCTGCCA GAGGCCAGAT GGTGCTTTAT ATGGAAGTTT ACATTTTGAC 1051 CCGGAGGCCT GCTCCTTCCG AGAAAGATTA TTAGAAGATG GCTACAATGT 1101 TTATCAAAGT GAAGCTCATG GTCTTCCTCT TCATCTTCCT GGGAACAAGT 1151 CGCCGCACCG GGACCCGGCG CCGCGGGGCC CGGCGCGCTT CCTCCCGCTG 1201 CCGGGCCTCC CGCCGGCGCT GCCGGAGCCG CCGGGCATCC TGGCGCCGCA 1251 GCCGCCGGAT GTTGGCTCCT CGGACCCGCT CTCCATGGTT GGTGGCAGCC 1301 AGGGCCGCTC GCCCAGCTAT GAAAGT SP1 - IgG1 Fc - Protein1 - FGF21 and 2 subunit complexes SEQ ID NO: 25 1 ATGGGCTGGA GCTGCATCAT CCTCTTCCTG GTGGCCACGG CCACTGGTGT 51 TCATTCAGAT AAAACTCACA CGTGCCCGCC GTGCCCGGCG CCGGAATTAC 101 TTGGTGGGCC CTCGGTCTTC CTCTTCCCGC CCAAGCCCA AGATACTTTA 151 ATGATCAGCC GCACGCCGGA GGTCACCTGT GTTGTTGTAG ATGTTTCTCA 201 TGAAGATCCT GAAGTAAAAT TTAACTGGTA TGTAGATGGT GTAGAAGTTC 251 ATAATGCAAA AACAAAACCA AGAGAAGAAC FATHERATTC CACCTACAGA 301 GTTGTTTCTG TTTTAACTGT TTTACATCAA GACTGGTTAA ATGGAAAAAGA 351 ATATAAATGT AAAGTTTCAA ATAAAGCCCT GCCGGCGCCC ATTGAAAAAAA 401 CCATCTCCAA AGCAAAAGGA CAGCCGCGGG AGCCCCAAGT TTACACGCTG 451 CCGCCGTCCA GAGATGATT AACAAAAAAT CAAGTTTCTC TCACCTGCCT 501 TGTAAAAGGA TTTTATCCTT CAGATATAGC AGTAGATGG GAATCAAATG 551 COMMUNITY AAATATTAT AAAACCACGC CGCCGGTTTT NATIONALITY 601 GGCTCCTTCT TCCTCTACAG CAAATTAACT GTAGATAAAA GCCGCTGGCA 651 ACAAGGAAAT GTCTTCTCCT GCAGTGTCAT GCATGAAGCA TTACATAATC 701 ATTACTCA AAAAAGTTTA TCTTTATCTC CTGGGAAAGG TGGTGGTGGC 751 AGTGGTGGTG GTGGCAGTGG TGGTGGTGGC AGCCACCCCA TCCCGGACAG 801 CAGCCCTTTA TTACAATTTG GTGGTCAGT AAGACAAAGA TATTATATA 851 INTERNATIONALGC WAVEWAVE RATE INTERNATIONAL FREQUENCY 901 ACTGTTGGTG GGGCGGCGGA CCAAAGTCCT GAAAGTTTAT RECORDAA 951 AGCATAAAA CCTGGTGTTA TTCAAATATT AGAGTAAAA ACCAGCCGCT 1001 TCCTCTGCCA AAGACCAGAT GGTGCTTTAT ATGGAAGTTT ACATTTTGAT 1051 CCTGAGGCCT GCTCCTTCAG AGAAAGATTA TTAGAAGATG GTTATAATGT 1101 TTATCAAAGT GAAGCACATG GTCTTCCTTT ACATTTACCT GGAAATAAAT 1151 CCCCGCACCG GGACCCGGCG CCGCGGGGCC CGGCGCGCTT CCTCCCGCTG 1201 CCGGGCCTCC CGCCGGCGCT GCCGGAGCCG CCGGGCATCC TGGCGCCGCA 1251 GCCGCCGGAT GTTGGCAGCA GTGACCCCCT CTCCATGGTT GGTGGCAGCC 1301 AAGGCCGCTC GCCCAGCTAT GAAAGT Coding sequence of SP1 - IgG1 Fc - linker1 - FGF21 variant 2 SEQ ID NO: 26 1 ATGGGCTGGA GCTGCATCAT CCTCTTCCTG GTGGCCACGG CCACTGGAGT 51 TCATTCAGAT AAAACCCACA CCTGCCCGCC CTGCCCGGCG CCGGAGCTGC 101 TGGGGGGCCC CAGTGTCTTC CTCTTCCCGC CCAAGCCCAA GGACACCCTC 151 ATGATTTCAA GAACTCCAGA AGTCACCTGT GTGGTGGTGG ATGTTTCTCA 201 TGAAGATCCT GAAGTAAAAT TTAACTGGTA TGTAGATGGT GTTGAAGTTC 251 ATAATGCCAA GACCAAGCCC CGGGAGGAGC AGTACAACAG CACCTACAGA 301 GTTGTTTCTG TTTTAACTGT TCTTCATCAG GACTGGTTAA ATGGAAAAGA 351 ATATAAATGT AAAGTTTCAA ATAAAGCCCT GCCGGCGCCC ATTGAAAAAA 401 CCATCTCCAA AGCAAAAGGA CAGCCCCGGG AGCCCCAGGT CTACACGCTG 451 CCGCCCAGCA GAGATGAATT AACAAAAAAT CAAGTCAGCC TCACCTGCCT 501 GGTGAAGGGC TTCTACCCCA GTGATATTGC TGTGGAGTGG GAGAGCAATG 551 GGCAGCCAGA AAATAATTAT AAAACCACCC CGCCGGTGCT GGACAGTGAT 601 GGCTCCTTCT TCCTCTACAG CAAATTAACT GTGGACAAGA GCCGCTGGCA 651 GCAAGGAAAT GTCTTCAGCT GCAGTGTCAT GCATGAAGCC CTGCACAACC 701 ACTACACCCA GAAGAGCCTC AGCCTCAGCC CTGGGAAAGG TGGTGGTGGC 751 AGTGGTGGTG GTGGCAGTGG TGGTGGTGGC AGCCACCCCA TCCCAGACAG 801 CAGCCCCCTG CTGCAGTTTG GTGGCCAGGT GCGGCAGCGC TACCTCTACA 851 CAGATGATGC CCAGCAGACA GAAGCACATT TAGAAATAAG AGAAGATGGA 901 ACTGTTGGTG GGGCGGCGGA CCAGAGCCCA GAAAGTTTAT TACAATTAAA 951 AGCATTAAAA CCTGGTGTCA TCCAGATTCT TGGAGTAAAA ACCAGCCGCT 1001 TCCTCTGCCA GAGGCCAGAT GGGGCCCTCT ATGGCAGCCT TCATTTTGAT 1051 CCTGAGGCCT GCAGCTTCCG GGAGCGGCTG CTGGAAGATG GCTACAATGT 1101 TTATCAAAGT GAAGCACATG GCCTGCCCCT GCACCTTCCT GGAAATAAAA 1151 GCCCCCACCG GGACCCGGCG CCCCGGGGCC CGGCGCGCTT CCTGCCCCTG 1201 CCTGGGCTGC CGCCGGCGCT GCCTGAGCCG CCGGGCATCC TGGCGCCCCA 1251 GCCCCCAGAT GTGGGCAGCA GTGACCCCCT CTCCATGGTT GGTGGCAGCC 1301 AGGGCCGCAG CCCCAGCTAT GAAAGT Coding sequence of SP1 - IgG1 Fc - linker1 - FGF21 variant 2 SEQ ID NO: 27 1 ATGGGCTGGA GCTGCATCAT CCTCTTCCTG GTGGCCACGG CCACTGGAGT 51 TCATTCAGAT AAAACTCACA CCTGCCCGCC CTGCCCGGCG CCGGAGCTGC 101 TGGGGGGCCC CAGTGTCTTC CTCTTCCCGC CCAAGCCCAA AGATACTTTA 151 ATGATTTCAA GAACTCCAGA AGTCACCTGT GTTGTTGTTG ATGTTTCTCA 201 TGAAGATCCT GAAGTAAAAT TTAACTGGTA TGTAGATGGT GTTGAAGTTC 251 ATAATGCCAA GACCAAGCCC CGGGAGGAGC AGTACAACAG CACCTACAGA 301 GTTGTTTCTG TTTTAACTGT TTTACATCAA GACTGGTTAA ATGGAAAAGA 351 ATATAAATGT AAAGTTTCAA ATAAAGCCCT GCCGGCGCCC ATTGAAAAAA 401 CCATCTCCAA AGCAAAAGGA CAGCCCCGGG AGCCCCAGGT CTACACGCTG 451 CCGCCCAGCA GAGATGAATT AACAAAAAAT CAAGTTTCTC TCACCTGCCT 501 GGTGAAAGGC TTCTACCCCA GTGATATTGC TGTGGAGTGG GAATCAAATG 551 GGCAGCCAGA AAATAATTAT AAAACCACGC CGCCGGTGCT GGACAGTGAT 601 GGCTCCTTCT TCCTCTACAG CAAATTAACT GTGGACAAGA GCCGCTGGCA 651 ACAAGGAAAT GTCTTCTCCT GCAGTGTCAT GCATGAAGCC CTGCACAACC 701 ACTACACCCA GAAGAGCCTC AGCCTCTCCC CGGGGAAAGG TGGTGGTGGC 751 AGTGGTGGTG GTGGCAGTGG TGGTGGTGGC AGCCACCCCA TCCCAGACAG 801 CAGCCCCCTG CTGCAGTTTG GTGGCCAGGT GCGGCAGCGC TACCTCTACA 851 CAGATGATGC CCAGCAAACA GAAGCACATT TAGAAATAAG AGAAGATGGA 901 ACTGTTGGTG GGGCGGCGGA CCAGTCTCCA GAAAGTTTAT TACAATTAAA 951 AGCATTAAAA CCTGGTGTCA TTCAAATATT AGGAGTAAAA ACCAGCCGCT 1001 TCCTCTGCCA GAGGCCAGAT GGTGCTTTAT ATGGAAGTTT ACATTTTGAT 1051 CCTGAGGCCT GCTCCTTCAG AGAAAGATTA TTAGAAGATG GCTACAATGT 1101 TTATCAAAGT GAAGCACATG GTCTTCCTCT TCATCTTCCT GGAAATAAAT 1151 CTCCCCACCG GGACCCGGCG CCGCGGGGCC CGGCGCGCTT CCTGCCCCTT 1201 CCTGGGCTGC CGCCGGCGCT GCCGGAGCCG CCGGGCATCC TGGCGCCCCA 1251 GCCGCCGGAT GTTGGCAGCA GTGACCCCCT CTCCATGGTT GGTGGCAGCC 1301 AGGGCCGCAG CCCCAGCTAT GAAAGT Coding sequence of SP1 - IgG1 Fc (YTE) - linker 1 - FGF21 variant 2 SEQ ID NO: 28 1 ATGGGCTGGA GCTGCATCAT CCTCTTCCTG GTGGCCACGG CCACTGGAGT 51 TCATTCAGAT AAAACCCACA CCTGCCCGCC CTGCCCGGCG CCGGAGCTGC 101 TGGGGGGCCC CAGTGTCTTC CTCTTCCCGC CCAAGCCCAA GGACACCCTC 151 TACATCACCC GGGAGCCAGA AGTCACCTGT GTGGTGGTGG ATGTTTCTCA 201 TGAAGATCCT GAGGTGAAGT TCAACTGGTA TGTTGATGGT GTTGAAGTTC 251 ATAATGCCAA GACCAAGCCC CGGGAGGAGC AGTACAACAG CACCTACAGA 301 GTTGTTTCTG TTTTAACTGT TCTTCATCAG GACTGGTTAA ATGGAAAAGA 351 ATATAAATGT AAAGTTTCAA ATAAAGCCCT GCCCGCGCCC ATTGAAAAAA 401 CCATCTCCAA AGCAAAAGGA CAGCCCCGGG AGCCCCAGGT GTACACGCTG 451 CCGCCCAGCA GAGATGAGCT GACCAAAAAT CAAGTCAGCC TCACCTGCCT 501 GGTGAAGGGC TTCTACCCCA GTGATATTGC TGTGGAGTGG GAGAGCAATG 551 GGCAGCCAGA AAATAATTAT AAAACCACCC CGCCGGTGCT GGACAGTGAT 601 GGCTCCTTCT TCCTCTACAG CAAATTAACT GTGGACAAGA GCCGCTGGCA 651 GCAAGGAAAT GTCTTCAGCT GCAGTGTCAT GCATGAAGCC CTGCACAACC 701 ACTACACCCA GAAAAGCCTC AGCCTCAGCC CTGGGAAAGG AGGTGGTGGC 751 AGTGGTGGTG GTGGCAGTGG TGGTGGTGGC AGCCACCCCA TCCCAGACAG 801 CAGCCCCCTG CTGCAGTTTG GTGGCCAGGT GCGGCAGCGC TACCTCTACA 851 CAGATGATGC CCAGCAGACA GAAGCTCATT TAGAAATAAG AGAAGATGGA 901 ACTGTTGGTG GTGCTGCTGA CCAGAGCCCT GAGAGCCTGC TGCAGCTGAA 951 GGCCCTGAAG CCAGGTGTCA TCCAGATTCT TGGAGTAAAA ACCAGCCGCT 1001 TCCTCTGCCA GAGGCCAGAT GGGGCCCTCT ATGGCAGCCT TCATTTTGAT 1051 CCTGAGGCCT GCAGCTTCCG GGAGCGGCTG CTGGAAGATG GCTACAATGT 1101 TTATCAAAGT GAAGCTCATG GCCTGCCCCT GCACCTTCCT GGAAATAAAA 1151 GCCCCCACAG AGATCCTGCC CCCCGGGGCC CTGCCCGCTT CCTGCCCCTG 1201 CCTGGGCTGC CCCCGGCCCT GCCTGAGCCG CCGGGCATCC TGGCCCCCCA 1251 GCCCCCAGAT GTGGGCAGCA GTGACCCCCT CTCCATGGTT GGTGGCAGCC 1301 AGGGCCGCAG CCCCAGCTAT GAAAGT Coding sequence of SP1 - IgG1 Fc (YTE) - linker1 - FGF21 variant 2 SEQ ID NO: 29 1 ATGGGCTGGA GCTGCATCAT CCTGTTCCTG GTGGCCACCG CCACCGGCGT 51 GCACAGCGAC AAGACCCACA CCTGCCCTCC TTGCCCTGCC CCTGAGCTGC 101 TGGGCGGCCC TAGCGTGTTC CTGTTCCCTC CTAAGCCTAA GGACACCCTG 从这里开始翻译,原内容中“151 TACATCACCA GAGAGCCTGA GGTGACCTGC GTGGTGGTGG ACGTGAGCCA”翻译为“151 TACATCACCA GAGAGCCTGA GGTGACCTGC GTGGTGGTGG ACGTGAGCCA”,这里原英文内容重复,可能有误,但按照要求保留原文。 201 CGAGGACCCT GAGGTGAAGT TCAACTGGTA CGTGGACGGC GTGGAGGTGC 251 ACAACGCCAA GACCAAGCCT AGAGAGGAGC AGTACAACAG CACCTACAGA 301 GTGGTGAGCG TGCTGACCGT GCTGCACCAG GACTGGCTGA ACGGCAAGGA 351 GTACAAGTGC AAGGTGAGCA ACAAGGCCCT GCCTGCCCCT ATCGAGAAGA 401 CCATCAGCAA GGCCAAGGGC CAGCCTAGAG AGCCTCAGGT GTACACCCTG 451 CCTCCTAGCA GAGACGAGCT GACCAAGAAC CAGGTGAGCC TGACCTGCCT 501 GGTGAAGGGC TTCTACCCTA GCGACATCGC CGTGGAGTGG GAGAGCAACG 551 GCCAGCCTGA GAACAACTAC AAGACCACCC CTCCTGTGCT GGACAGCGAC 601 GGCAGCTTCT TCCTGTACAG CAAGCTGACC GTGGACAAGA GCAGATGGCA 651 GCAGGGCAAC GTGTTCAGCT GCAGCGTGAT GCACGAGGCC CTGCACAACC 701 ACTACACCCA GAAAAGCCTG AGCCTGAGCC CTGGCAAGGG CGGCGGCGGC 751 AGCGGCGGCG GCGGCAGCGG CGGCGGCGGC AGCCACCCTA TCCCTGACAG 801 CAGCCCTCTG CTGCAGTTCG GCGGCCAGGT GAGACAGAGA TACCTGTACA 851 CCGACGACGC CCAGCAGACC GAGGCCCACC TGGAGATCAG AGAGGACGGC 901 ACCGTGGGCG GCGCCGCCGA CCAGAGCCCT GAGAGCCTGC TGCAGCTGAA 951 GGCCCTGAAG CCTGGCGTGA TCCAGATCCT GGGCGTGAAG ACCAGCAGAT 1001 TCCTGTGCCA GAGACCTGAC GGCGCCCTGT ACGGCAGCCT GCACTTCGAC 1051 CCTGAGGCCT GCAGCTTCAG AGAGAGACTG CTGGAGGACG GCTACAACGT 1101 GTACCAGAGC GAGGCCCACG GCCTGCCTCT GCACCTGCCT GGCAACAAGA 1151 GCCCTCACAG AGACCCTGCC CCTAGAGGCC CTGCCAGATT CCTGCCTCTG 1201 CCTGGCCTGC CTCCTGCCCT GCCTGAGCCT CCTGGCATCC TGGCCCCTCA 1251 GCCTCCTGAC GTGGGCAGCA GCGACCCTCT GAGCATGGTG GGCGGCAGCC 1301 AGGGCAGAAG CCCTAGCTAC GAGAGC Coding sequence of SP1 - IgG1 Fc (YTE) - linker1 - FGF Variant 2 SEQ ID NO: 30 1 ATGGGCTGGA GCTGCATCAT CCTCTTCCTG GTGGCCACAG CCACCGGTGT 51 CCACAGCGAC AAAACCCACA CCTGTCCTCC CTGCCCCGCC CCAGAGCTGC 101 TGGGCGGCCC CTCTGTGTTT CTGTTCCCCC CAAAGCCCAA GGACACCCTG 151 TACATCACCC GAGAGCCAGA AGTTACTTGT GTGGTGGTTG ATGTCAGCCA 201 TGAAGACCCC GAGGTGAAGT TCAACTGGTA CGTGGATGGT GTGGAAGTGC 251 ACAATGCCAA GACAAAGCCA AGAGAGGAGC AGTACAACAG CACCTACAGA 301 GTGGTCTCTG TCCTGACAGT GCTCCACCAG GACTGGCTGA ATGGCAAGGA 351 GTACAAGTGC AAGGTGAGCA ACAAAGCCTT GCCTGCCCCC ATTGAGAAGA 401 CCATCTCCAA GGCCAAAGGC CAGCCCAGGG AGCCCCAGGT CTACACACTG 451 CCACCCAGCA GAGATGAACT GACCAAGAAC CAGGTGTCCC TCACCTGCCT 501 GGTCAAGGGC TTCTACCCCA GTGACATCGC CGTGGAGTGG GAGAGCAATG 551 GACAGCCTGA GAACAACTAC AAAACAACAC CACCTGTGCT GGACAGTGAT 601 GGAAGCTTCT TCCTCTACAG CAAGCTGACT GTGGACAAGA GCCGCTGGCA 651 GCAAGGAAAT GTCTTCTCCT GCAGCGTGAT GCACGAGGCT CTGCACAACC 701 ACTACACCCA GAAGTCCCTG AGCCTGTCTC CAGGAAAAGG AGGCGGGGGC 751 AGTGGTGGAG GTGGCTCTGG GGGTGGTGGG AGCCACCCCA TCCCAGACAG 801 CAGCCCACTG CTCCAGTTTG GAGGGCAGGT TCGGCAGCGC TACCTGTATA 851 CAGATGATGC CCAGCAGACA GAAGCCCACC TGGAGATCCG GGAGGACGGC 901 ACCGTGGGAG GAGCTGCTGA CCAGAGCCCT GAAAGCCTCC TGCAGCTGAA 951 AGCACTGAAG CCTGGGGTCA TCCAGATCCT GGGGGTGAAA ACTTCCCGCT 1001 TCCTGTGCCA GAGGCCTGAT GGGGCCCTCT ATGGAAGTCT TCATTTTGAT 1051 CCTGAGGCCT GCTCCTTCAG AGAAAGACTT CTGGAAGATG GCTACAATGT 1101 GTACCAGAGT GAAGCTCATG GCCTGCCCCT GCATCTGCCT GGCAACAAGT 1151 CTCCCCACCG GGACCCAGCT CCCCGGGGCC CTGCTAGATT TCTTCCTCTT 1201 CCAGGCCTCC CTCCAGCCCT GCCGGAGCCT CCTGGAATTC TGGCCCCTCA 1251 GCCACCAGAT GTGGGCTCCT CTGACCCTCT GTCCATGGTG GGGGGCAGCC 1301 AGGGCCGCAG CCCCTCCTAT GAAAGT Coding sequence of SP1 - IgG1 Fc (YTE) - linker 1 - FGF21 variant 2 SEQ ID NO: 31 1 ATGGGCTGGA GCTGCATCAT CCTCTTCCTG GTGGCCACAG CCACCGGTGT 51 CCACAGCGAC AAAACCCACA CCTGTCCTCC CTGCCCCGCC CCAGAGCTGC 101 TGGGCGGCCC CTCTGTGTTT CTGTTCCCCC CAAAGCCCAA GGACACCCTG 151 TACATCACCC GAGAGCCAGA AGTTACTTGT GTGGTGGTTG ATGTCAGCCA 201 TGAAGACCCC GAGGTGAAGT TCAACTGGTA CGTGGATGGT GTGGAAGTGC 251 ACAATGCCAA GACAAAGCCA AGAGAGGAGC AGTACAACAG CACCTACAGA 301 GTGGTCTCTG TCCTGACAGT GCTCCACCAG GACTGGCTGA ATGGCAAGGA 351 GTACAAGTGC AAGGTGAGCA ACAAAGCCTT GCCTGCCCCC ATTGAGAAGA 401 CCATCTCCAA GGCCAAAGGC CAGCCCAGGG AGCCCCAGGT CTACACACTG 451 CCACCCAGCA GAGATGAACT GACCAAGAAC CAGGTGTCCC TCACCTGCCT 501 GGTCAAGGGC TTCTACCCCA GTGACATCGC CGTGGAGTGG GAGAGCAATG 551 GACAGCCTGA GAACAACTAC AAAACAACAC CACCTGTGCT GGACAGTGAT 601 GGAAGCTTCT TCCTCTACAG CAAGCTGACT GTGGACAAGA GCCGCTGGCA 651 GCAAGGAAAT GTCTTCTCCT GCAGCGTGAT GCACGAGGCT CTGCACAACC 701 ACTACACCCA GAAGTCCCTG AGCCTGTCTC CAGGAAAAGG AGGTGGAGGC 751 AGTGGTGGCG GGGGCTCTGG GGGTGGTGGG AGCCACCCCA TCCCAGACAG 801 CAGCCCACTG CTCCAGTTTG GAGGGCAGGT TCGGCAGCGC TACCTGTATA 851 CAGATGATGC CCAGCAGACA GAAGCCCACC TGGAGATCCG GGAGGACGGC 901 ACCGTGGGAG GAGCTGCTGA CCAGAGCCCT GAAAGCCTCC TGCAGCTGAA 951 AGCACTGAAG CCTGGGGTCA TCCAGATCCT GGGGGTGAAA ACTTCCCGCT 1001 TCCTGTGCCA GAGGCCTGAT GGGGCCCTCT ATGGAAGTCT TCATTTTGAT 1051 CCTGAGGCCT GCTCCTTCAG AGAAAGACTT CTGGAAGATG GCTACAATGT 1101 GTACCAGAGT GAAGCTCATG GCCTGCCCCT GCATCTGCCT GGCAACAAGT 1151 CTCCCCACCG GGACCCAGCT CCCCGGGGCC CTGCTAGATT TCTTCCTCTT 1201 CCAGGCCTCC CTCCAGCCCT GCCGGAGCCT CCTGGAATTC TGGCCCCTCA 1251 GCCACCAGAT GTGGGCTCCT CTGACCCTCT GTCCATGGTG GGGGGCAGCC 1301 AGGGCCGCAG CCCCTCCTAT GAAAGT Coding sequence of SP1 - IgG1 Fc (YTE) - linker 1 - FGF21 variant 2 SEQ ID NO: 32 1 ATGGGCTGGA GCTGCATCAT CCTCTTCCTG GTGGCCACAG CCACCGGTGT 51 CCACAGCGAC AAAACCCACA CCTGTCCTCC CTGCCCCGCC CCAGAGCTGC 101 TGGGCGGCCC CTCTGTGTTT CTGTTCCCCC CAAAGCCCAA GGACACCCTG 151 TACATCACCC GAGAGCCAGA AGTTACTTGT GTGGTGGTTG ATGTCAGCCA 201 TGAAGACCCC GAGGTGAAGT TCAACTGGTA CGTGGATGGT GTGGAAGTGC 251 ACAATGCCAA GACAAAGCCA AGAGAGGAGC AGTACAACAG CACCTACAGA 301 GTGGTCTCTG TCCTGACAGT GCTCCACCAG GACTGGCTGA ATGGCAAGGA 351 GTACAAGTGC AAGGTGAGCA ACAAAGCCTT GCCTGCCCCC ATTGAGAAGA 401 CCATCTCCAA GGCCAAAGGC CAGCCCAGGG AGCCCCAGGT CTACACACTG 451 CCACCCAGCA GAGATGAACT GACCAAGAAC CAGGTGTCCC TCACCTGCCT 501 GGTCAAGGGC TTCTACCCCA GTGACATCGC CGTGGAGTGG GAGAGCAATG 551 GACAGCCTGA GAACAACTAC AAAACAACAC CACCTGTGCT GGACAGTGAT 601 GGAAGCTTCT TCCTCTACAG CAAGCTGACT GTGGACAAGA GCCGCTGGCA 651 GCAAGGAAAT GTCTTCTCCT GCAGCGTGAT GCACGAGGCT CTGCACAACC 701 ACTACACCCA GAAGTCCCTG AGCCTGTCTC CAGGAAAAGG AGGCGGGGGC 751 TCTGGGGGTG GTGGCAGTGG TGGAGGTGGG AGCCACCCCA TCCCAGACAG 801 CAGCCCACTG CTCCAGTTTG GAGGGCAGGT TCGGCAGCGC TACCTGTATA 851 CAGATGATGC CCAGCAGACA GAAGCCCACC TGGAGATCCG GGAGGACGGC 901 ACCGTGGGAG GAGCTGCTGA CCAGAGCCCT GAAAGCCTCC TGCAGCTGAA 951 AGCACTGAAG CCTGGGGTCA TCCAGATCCT GGGGGTGAAA ACTTCCCGCT 1001 TCCTGTGCCA GAGGCCTGAT GGGGCCCTCT ATGGAAGTCT TCATTTTGAT 1051 CCTGAGGCCT GCTCCTTCAG AGAAAGACTT CTGGAAGATG GCTACAATGT 1101 GTACCAGAGT GAAGCTCATG GCCTGCCCCT GCATCTGCCT GGCAACAAGT 1151 CTCCCCACCG GGACCCAGCT CCCCGGGGCC CTGCTAGATT TCTTCCTCTT 1201 CCAGGCCTCC CTCCAGCCCT GCCGGAGCCT CCTGGAATTC TGGCCCCTCA 1251 GCCACCAGAT GTGGGCTCCT CTGACCCTCT GTCCATGGTG GGGGGCAGCC 1301 AGGGCCGCAG CCCCTCCTAT GAAAGT Coding sequence of SP1 - IgG1 Fc (YTE) - linker 1 - FGF21 variant 2 SEQ ID NO: 33 1 ATGGGCTGGA GCTGCATCAT CCTCTTCCTG GTGGCCACAG CCACCGGTGT 51 CCACAGCGAC AAAACCCACA CCTGTCCTCC CTGCCCCGCC CCAGAGCTGC 101 TGGGCGGCCC CTCTGTGTTT CTGTTCCCCC CAAAGCCCAA GGACACCCTG 151 TACATCACCC GAGAGCCAGA AGTTACTTGT GTGGTGGTTG ATGTCAGCCA 201 TGAAGACCCC GAGGTGAAGT TCAACTGGTA CGTGGATGGT GTGGAAGTGC 251 ACAATGCCAA GACAAAGCCA AGAGAGGAGC AGTACAACAG CACCTACAGA 301 GTGGTCTCTG TCCTGACAGT GCTCCACCAG GACTGGCTGA ATGGCAAGGA 351 GTACAAGTGC AAGGTGAGCA ACAAAGCCTT GCCTGCCCCC ATTGAGAAGA 401 CCATCTCCAA GGCCAAAGGC CAGCCCAGGG AGCCCCAGGT CTACACACTG 451 CCACCCAGCA GAGATGAACT GACCAAGAAC CAGGTGTCCC TCACCTGCCT 501 GGTCAAGGGC TTCTACCCCA GTGACATCGC CGTGGAGTGG GAGAGCAATG 551 GACAGCCTGA GAACAACTAC AAAACAACAC CACCTGTGCT GGACAGTGAT 601 GGAAGCTTCT TCCTCTACAG CAAGCTGACT GTGGACAAGA GCCGCTGGCA 651 GCAAGGAAAT GTCTTCTCCT GCAGCGTGAT GCACGAGGCT CTGCACAACC 701 ACTACACCCA GAAGTCCCTG AGCCTGTCTC CAGGAAAAGG AGGTGGAGGC 751 TCTGGGGGTG GTGGCAGTGG TGGGGGCGGG AGCCACCCCA TCCCAGACAG 801 CAGCCCACTG CTCCAGTTTG GAGGGCAGGT TCGGCAGCGC TACCTGTATA 851 CAGATGATGC CCAGCAGACA GAAGCCCACC TGGAGATCCG GGAGGACGGC 901 ACCGTGGGAG GAGCTGCTGA CCAGAGCCCT GAAAGCCTCC TGCAGCTGAA 951 AGCACTGAAG CCTGGGGTCA TCCAGATCCT GGGGGTGAAA ACTTCCCGCT 1001 TCCTGTGCCA GAGGCCTGAT GGGGCCCTCT ATGGAAGTCT TCATTTTGAT 1051 CCTGAGGCCT GCTCCTTCAG AGAAAGACTT CTGGAAGATG GCTACAATGT 1101 GTACCAGAGT GAAGCTCATG GCCTGCCCCT GCATCTGCCT GGCAACAAGT 1151 CTCCCCACCG GGACCCAGCT CCCCGGGGCC CTGCTAGATT TCTTCCTCTT 1201 CCAGGCCTCC CTCCAGCCCT GCCGGAGCCT CCTGGAATTC TGGCCCCTCA 1251 GCCACCAGAT GTGGGCTCCT CTGACCCTCT GTCCATGGTG GGGGGCAGCC 1301 AGGGCCGCAG CCCCTCCTAT GAAAGT Coding sequence of SP1 - IgG1 Fc (YTE) - linker 1 - FGF21 variant 2 SEQ ID NO: 34 1 ATGGGCTGGA GCTGCATCAT CCTCTTCCTG GTGGCCACAG CCACCGGTGT 51 CCACAGCGAC AAAACCCACA CCTGTCCTCC CTGCCCCGCC CCAGAGCTGC 101 TGGGCGGCCC CTCTGTGTTT CTGTTCCCCC CAAAGCCCAA GGACACCCTG 151 TACATCACCC GAGAGCCAGA AGTTACTTGT GTGGTGGTTG ATGTCAGCCA 201 TGAAGACCCC GAGGTGAAGT TCAACTGGTA CGTGGATGGT GTGGAAGTGC 251 ACAATGCCAA GACAAAGCCA AGAGAGGAGC AGTACAACAG CACCTACAGA 301 GTGGTCTCTG TCCTGACAGT GCTCCACCAG GACTGGCTGA ATGGCAAGGA 351 GTACAAGTGC AAGGTGAGCA ACAAAGCCTT GCCTGCCCCC ATTGAGAAGA 401 CCATCTCCAA GGCCAAAGGC CAGCCCAGGG AGCCCCAGGT CTACACACTG 451 CCACCCAGCA GAGATGAACT GACCAAGAAC CAGGTGTCCC TCACCTGCCT 501 GGTCAAGGGC TTCTACCCCA GTGACATCGC CGTGGAGTGG GAGAGCAATG 551 GACAGCCTGA GAACAACTAC AAAACAACAC CACCTGTGCT GGACAGTGAT 601 GGAAGCTTCT TCCTCTACAG CAAGCTGACT GTGGACAAGA GCCGCTGGCA 651 GCAAGGAAAT GTCTTCTCCT GCAGCGTGAT GCACGAGGCT CTGCACAACC 701 ACTACACCCA GAAGTCCCTG AGCCTGTCTC CAGGAAAAGG AGGTGGGGGA 751 AGTGGTGGTG GAGGCAGTGG GGGTGGCGGG AGCCACCCCA TCCCAGACAG 801 CAGCCCACTG CTCCAGTTTG GAGGGCAGGT TCGGCAGCGC TACCTGTATA 851 CAGATGATGC CCAGCAGACA GAAGCCCACC TGGAGATCCG GGAGGACGGC 901 ACCGTGGGAG GAGCTGCTGA CCAGAGCCCT GAAAGCCTCC TGCAGCTGAA 951 AGCACTGAAG CCTGGGGTCA TCCAGATCCT GGGGGTGAAA ACTTCCCGCT 1001 TCCTGTGCCA GAGGCCTGAT GGGGCCCTCT ATGGCTCTCT TCATTTTGAT 1051 CCTGAGGCCT GCTCCTTCAG AGAAAGACTT CTGGAAGATG GCTACAATGT 1101 GTACCAGAGT GAAGCTCATG GCCTGCCCCT GCATCTGCCT GGCAACAAGT 1151 CTCCCCACCG GGACCCAGCT CCCCGGGGCC CTGCTAGATT TCTTCCTCTT 1201 CCAGGCCTCC CTCCAGCCCT GCCGGAGCCT CCTGGAATTC TGGCCCCTCA 1251 GCCACCAGAT GTGGGCTCCT CTGACCCTCT GTCCATGGTG GGGGGCAGCC 1301 AGGGCCGCAG CCCCTCCTAT GAAAGT Coding sequence of SP2 - IgG1 Fc - linker4 - FGF21 variant 3 SEQ ID NO: 35 1 ATGGAGACCG ACACCCTGCT CCTGATGGTG CTCCTGCTGT GGGTGCCCGG 51 CTCCACCGGC GATAAGACCC ATACCTGCCC CCCTTGCCCC GCCCCCGAGC 101 TCCTGGGCGG CCCTAGCGTG TTCCTGTTCC CCCCCAAGCC CAAGGACACC 151 CTGATGATCA GCAGAACCCC CGAGGTGACC TGCGTGGTCG TGGACGTGAG 201 CCACGAGGAC CCCGAGGTGA AGTTCAACTG GTACGTGGAC GGCGTGGAGG 251 TGCACAACGC CAAGACCAAG CCTAGAGAGG AGCAGTACAA CAGCACCTAC 301 AGAGTGGTGA GCGTGCTGAC CGTGCTGCAC CAAGACTGGC TGAACGGCAA 351 GGAGTACAAG TGCAAGGTGA GCAACAAGGC CCTGCCCGCC CCCATCGAGA 401 AGACCATCAG CAAGGCCAAG GGGCAGCCTA GAGAGCCCCA AGTGTACACC 451 CTGCCCCCTA GCAGAGACGA GCTGACCAAG AACCAAGTGA GCCTGACCTG 501 CCTGGTGAAG GGCTTCTACC CTAGCGACAT CGCCGTGGAG TGGGAGAGCA 551 ACGGGCAGCC CGAGAACAAC TACAAGACCA CCCCCCCCGT GCTGGACAGC 601 GACGGCAGCT TCTTCCTGTA CAGCAAGCTG ACCGTGGACA AGAGCAGATG 651 GCAGCAAGGC AACGTGTTCA GCTGCAGCGT GATGCACGAG GCCCTGCACA 701 ACCACTACAC ACAGAAGTCC CTGAGCCTGA GCCCTGGCAA AGGGGGGGGC 751 GGGGGGAGCG GCGGGGGCAG CGGGGGGGGC GGGAGCCACC CTATCCCCGA 801 CAGCAGCCCC CTGCTGCAGT TCGGCGGCCA AGTGAGACAG AGATACCTGT 851 ACACCGACGA CGCTCAGCAG ACAGAGGCCC ATCTGGAAAT CAGAGAGGAC 901 GGGACCGTGG GCGGGGCTGC CGATCAGAGC CCCGAGAGCC TGCTGCAGCT 951 GAAGGCCCTG AAGCCCGGCG TGATTCAGAT CCTGGGCGTG AAGACAAGCA 1001 GATTCCTGTG TCAGAGACCC GACGGCGCCC TGTACGGCAG CCTGCACTTC 1051 GACCCCGAGG CCTGCAGCTT CAGAGAGAGA CTGCTGGAGG ACGGCTACAA 1101 CGTGTATCAG AGCGAGGCCC ACGGCCTGCC CCTCCACCTG CCCGGGAACA 1151 AGAGCCCTCA CAGAGATCCT GCTCCTAGAG GCCCCGCTAG ATTTCTCCCC 1201 CTGCCCGGGC TGCCTCCCGC CCCTCCTGAA CCCCCTGGCA TCCTCGCTCC 1251 CCAACCCCCT GATGTGGGCA GCAGCGACCC CCTGAGCATG GTGGGGGGCA 1301 GCCAAGGCAG AAGCCCTAGC TACGCTAGC Coding sequence of SP1 - ABD - linker2 - FGF21 variant 2 SEQ ID NO: 36 1 ATGGGCTGGA GCTGCATCAT CCTCTTCCTG GTGGCCACAG CCACTGGAGT 51 TCATTCTGCA GAGGCCAAGG AGGCTGCCAA TGCAGAGCTG GACAGCTATG 101 GTGTCAGTGA CTTCTACAAG CGCCTCATTG ACAAGGCCAA GACTGTGGAG 151 GGTGTGGAGG CCCTGAAGGA TGCCATCCTG GCGGCGCTGC CTGGGGGCGG 201 CGGCAGTGGT GGTGGCAGCC ACCCCATCCC AGACAGCAGC CCCCTGCTGC 251 AGTTTGGTGG CCAGGTGCGG CAGCGCTACC TCTACACAGA TGATGCCCAG 301 CGACAGAGG CCCACCTGGA GATCAGAGAA GATGGAACAG TTGGAGGAGC 351 TGCTGACCAG AGCCCTGAGA GCCTGCTGCA GCTGAAGGCC CTGAAGCCTG 401 GGGTCATCCA GATCCTGGGC GTGAAGACCA GCCGCTTCCT CTGCCAGAGG 451 CCAGATGGGG CCCTGTATGG CAGCCTGCAC TTTGATCCTG AGGCCTGCTC 501 CTTCCGGGAG CGGCTGCTGG GGGGCTA CAATGTCTAC CAGAGTGAAG 551 CTCATGGCCT GCCCCTGCAC CTGCCTGGGA ACAAGAGCCC CCACCGGGAC 601 CCTGCCCCCC GGGGCCCTGC CCGCTTCCTG CCCCTGCCTG GGCTGCCCCC 651 AGCCCTGCCT GAGCCCCCAG GCATCCTGGC CCCCCAGCCC CCAGATGTGG 701 GCAGCAGTGA CCCCCTGTCC ATGGTGGGCG GCAGCCAGGG CCGCAGCCCC 751 AGCTATGAGA GC SP1 - ABD - PAR2 - FGF21 and 2 subunit genes SEQ ID NO: 37 1 ATGGGCTGGA GCTGCATCAT CCTCTTCCTG GTGGCCACGG CCACTGGAGT 51 TCATTCAGCA GAAGCAAAAG AAGCAGCAAA TGCAGAGCTG GACAGCTATG 101 GTGTTTCAGA TTTTTATAAA AGATTAATTG ATAAAGCAAA AACTGTAGAA 151 GGTGTGGAGG CCTTGAAAGA TGCCATCCTG GCGGCGCTGC CTGGTGGTGG 201 TGGCAGTGGT GGTGGCAGCC ACCCCATCCC AGACAGCAGC CCCCTGCTGC 251 AGTTTGGTGG CCAGGTGCGG CAGCGCTACC TCTACACAGA TGATGCCCAG 301 CAGACAGAAG CACATTTAGA AATAAGAGAA GATGGAACTG TTGGTGGTGC 351 TGCTGACCAG AGCCCTGAGA GCCTGCTGCA GCTGAAGGCC CTGAAGCCAG 401 GTGTCATCCA GATTCTTGGT GTGAAGACCA GCCGCTTCCT CTGCCAGAGG 451 CCAGATGGGG CCCTCTATGG CAGCCTTCAT TTTGATCCTG AGGCCTGCAG 501 CTTCCGGGAG CGGCTGCTGG AAGATGGCTA CAATGTTTAT CAAAGTGAAG 551 CTCATGGCCT GCCCCTGCAC CTTCCTGGAA ATAAAAGCCC CCACAGAGAT 601 CCTGCCCCCC GGGGCCCTGC CCGCTTCCTG CCCCTGCCTG GGCTGCCCCC 651 GGCCCTGCCT GAGCCGCCGG GCATCCTGGC CCCCCAGCCC CCAGATGTGG 701 GCAGCAGTGA CCCCCTCTCC ATGGTTGGTG GCAGCCAGGG CCGCAGCCCC751 AGCTATGAAA GT Coding sequence of SP1 - ABD - linker2 - FGF21 variant 2 SEQ ID NO: 38 1 ATGGGCTGGA GCTGCATCAT CCTCTTCCTG GTGGCCACGG CCACTGGAGT 51 TCATTCTGCA GAGGCCAAGG AGGCGGCCAA TGCAGAGCTG GACAGCTATG 101 GTGTCAGTGA CTTCTACAAG CGCCTCATTG ACAAGGCCAA GACAGTAGAA 151 GGTGTGGAGG CCCTGAAGGA TGCCATCCTG GCGGCGCTGC CTGGTGGTGG 201 TGGCAGTGGT GGTGGCAGCC ACCCCATCCC AGACAGCAGC CCCCTGCTGC 251 AGTTTGGTGG CCAGGTGCGG CAGCGCTACC TCTACACAGA TGATGCCCAG 301 CGACAGAGG CCCACCTGGA GATCAGAGAA GATGGAACAG TTGGTGGTGC 351 TGCTGACCAG AGCCCTGAGA GCCTGCTGCA GCTGAAGGCC CTGAAGCCTG 401 GTGTCATCCA GATCCTGGGT GTGAAGACCA GCCGCTTCCT CTGCCAGAGG 451 CCAGATGGGG CCCTCTATGG CAGCCTGCAC TTTGATCCTG AGGCCTGCTC 501 CTTCCGGGAG CGGCTGCTGG GGGGCTA CAATGTCTAC CAGAGTGAAG 551 CTCATGGCCT GCCCCTGCAC CTGCCTGGGA ACAAGAGCCC CCACCGGGAC 601 CCCGCGCCCC GGGGCCCTGC CCGCTTCCTG CCCCTGCCTG GGCTGCCGCC 651 GGCCCTGCCT GAGCCGCCGG GCATCCTGGC CCCCCAGCCC CCAGATGTGG 701 GCAGCAGTGA CCCCCCTCTCC ATGGTGGGCG GCAGCCAGGG CCGCAGCCCC 751 AGCTATGAAA GT Coding sequence of SP1 - ABD - linker 2 - FGF21 variant 2 SEQ ID NO: 39 1 ATGGGCTGGA GCTGCATCAT CCTCTTCCTG GTGGCCACAG CCACTGGAGT 51 TCATTCAGCA GAGGCCAAAG AAGCAGCAAA TGCAGAGCTG GACAGCTATG 101 GTGTCAGTGA CTTCTACAAG CGCCTCATTG ATAAAGCAAA AACTGTGGAA 151 GGTGTGGAGG CCCTGAAAGA TGCCATCCTG GCGGCGCTGC CTGGTGGTGG 201 TGGCAGTGGT GGTGGCAGCC ACCCCATCCC AGACAGCAGC CCCCTGCTGC 251 AGTTTGGTGG CCAGGTGCGG CAGCGCTACC TCTACACAGA TGATGCCCAG 301 CAGACAGAAG CCCACCTGGA AATAAGAGAA GATGGAACTG TTGGTGGTGC 351 TGCTGACCAG AGCCCTGAGA GCCTGCTGCA GCTGAAGGCC CTGAAGCCAG 401 GTGTCATCCA GATTCTTGGT GTGAAGACCA GCCGCTTCCT CTGCCAGAGG 451 CCAGATGGGG CCCTCTATGG CAGCCTGCAC TTTGATCCTG AGGCCTGCAG 501 CTTCCGGGAG CGGCTGCTGG AAGATGGCTA CAATGTCTAC CAGAGTGAAG 551 CTCATGGCCT GCCCCTGCAC CTGCCTGGAA ATAAAAGCCC CCACAGAGAT 601 CCTGCCCCCC GGGGCCCTGC CCGCTTCCTG CCCCTGCCTG GGCTGCCCCC 651 AGCCCTGCCT GAGCCCCCGG GCATCCTGGC CCCCCAGCCC CCAGATGTGG 701 GCAGCAGTGA CCCCCTCTCC ATGGTGGGGG GCAGCCAGGG CCGCAGCCCC 751 AGCTATGAAA GT Coding sequence of SP1 - ABD - linker 2 - FGF21 variant 2 SEQ ID NO: 40 1 ATGGGCTGGA GCTGCATCAT CCTGTTCCTG GTGGCCACCG CCACCGGCGT 51 GCACAGCGCC GAGGCCAAGG AGGCCGCCAA CGCCGAGCTG GACAGCTACG 101 GCGTGAGCGA CTTCTACAAG AGACTGATCG ACAAGGCCAA GACCGTGGAG 151 GGCGTGGAGG CCCTGAAGGA CGCCATCCTG GCCGCCCTGC CTGGCGGCGG 201 CGGCAGCGGC GGCGGCAGCC ACCCTATCCC TGACAGCAGC CCTCTGCTGC 251 AGTTCGGCGG CCAGGTGAGA CAGAGATACC TGTACACCGA CGACGCCCAG 301 CAGACCGAGG CCCACCTGGA GATCAGAGAG GACGGCACCG TGGGCGGCGC 351 CGCCGACCAG AGCCCTGAGA GCCTGCTGCA GCTGAAGGCC CTGAAGCCTG 401 GCGTGATCCA GATCCTGGGC GTGAAGACCA GCAGATTCCT GTGCCAGAGA 451 CCTGACGGCG CCCTGTACGG CAGCCTGCAC TTCGACCCTG AGGCCTGCAG 501 CTTCAGAGAG AGACTGCTGG AGGACGGCTA CAACGTGTAC CAGAGCGAGG 551 CCCACGGCCT GCCTCTGCAC CTGCCTGGCA ACAAGAGCCC TCACAGAGAC 601 CCTGCCCCTA GAGGCCCTGC CAGATTCCTG CCTCTGCCTG GCCTGCCTCC 651 TGCCCTGCCT GAGCCTCCTG GCATCCTGGC CCCTCAGCCT CCTGACGTGG 701 GCAGCAGCGA CCCTCTGAGC ATGGTGGGCG GCAGCCAGGG CAGAAGCCCT 751 AGCTACGAGA GC Coding sequence of SP1-ABD-Linker2-FGF21 variant 2 SEQ ID NO: 41 1 ATGGGCTGGA GCTGCATCAT CCTCTTCCTG GTGGCCACAG CCACTGGAGT 51 TCATTCAGCA GAGGCCAAGG AAGCTGCCAA TGCAGAGCTG GACAGCTATG 101 GAGTTTCTGA CTTCTACAAG CGCCTCATTG ATAAAGCCAA GACTGTGGAA 151 GGTGTGGAGG CCCTGAAAGA TGCCATCCTG GCTGCCCTGC CTGGAGGAGG 201 AGGAAGTGGT GGTGGCAGCC ACCCCATCCC AGACAGCAGC CCCCTGCTGC 251 AGTTTGGGGG CCAGGTGCGG CAGCGCTACC TCTACACAGA TGATGCCCAG 301 CAGACAGAGG CCCACCTGGA GATCAGAGAA GATGGAACAG TTGGAGGAGC 351 TGCTGACCAG AGCCCTGAGA GCCTGCTGCA GCTGAAGGCC CTGAAGCCTG 401 GGGTCATCCA GATCCTGGGC GTGAAGACCA GCCGCTTCCT CTGCCAGAGG 451 CCAGATGGGG CCCTGTATGG CAGCCTGCAC TTTGATCCTG AGGCCTGCAG 501 CTTCCGGGAG CGGCTGCTGG AAGATGGCTA CAATGTCTAC CAGAGTGAGG 551 CCCATGGCCT GCCCCTGCAC CTGCCTGGAA ATAAAAGCCC CCACAGAGAT 601 CCTGCCCCCC GGGGCCCTGC CCGCTTCCTG CCCCTGCCTG GCCTGCCCCC 651 AGCCCTGCCT GAGCCCCCAG GCATCCTGGC CCCCCAGCCC CCAGATGTGG 701 GCAGCAGTGA CCCCCTGTCC ATGGTGGGCG GCAGCCAGGG CCGCAGCCCC 751 AGCTATGAAA GT Coding sequence of SP1 - anti - albumin VHH - linker 2 - FGF21 variant 2 SEQ ID NO: 42 1 ATGGGCTGGA GCTGCATCAT CCTCTTCCTG GTGGCCACAG CCACTGGAGT 51 CCACAGTGAG GTGCAGCTGG TGGAGAGTGG TGGTGGCCTG GTGCAGCCTG 101 GGAACAGCCT GCGGCTGAGC TGTGCTGCCA GTGGCTTCAC CTTCAGCAGC 151 TTTGGAATGT CCTGGGTGCG GCAGGCCCCT GGGAAGGGCC TGGAGTGGGT 201 CAGCAGCATC AGTGGCAGTG GCAGTGACAC CCTCTATGCA GATTCTGTGA 251 AGGGCCGCTT CACCATCAGC CGGGACAATG CCAAGACCAC CCTCTACCTG 301 ACTIVITY GCCTGCGGCC ACTIVITY GCTGTGTACT ACTGCACCAT 351 TGGTGGCAGC CTCAGCCGCA GCAGCCAGGG CACCCTGGTG ACTGTCAGCA 401 GTGGTGGTGG TGGCAGTGGT GGTGGCAGCC ACCCCATCCC AGACAGCAGC 451 CCCCTGCTGC AGTTTGGTGG CCAGGTGCGG CAGCGCTACC TCTACACAGA 501 TGATGCCCAG CAGACAGAGG CCCACCTGGA GATCAGAGAA GATGGAACAG 551 TTGGTGGTGC TGCTGACCAG AGCCCTGAGA GCCTGCTGCA GCTGAAGGCC 601 CTGAAGCCTG GGGTCATCCA GATCCTGGGC GTGAAGCCA GCCGCTTCCT 651 CTGCCAGAGG CCAGATGGGG CCCTGTATGG CAGCCTGCAC TTTGATCCTG 701 AGGCCTGCTC CTTCCGGGAG CGGCTGCTGG AGGATGGCTA CAATGTCTAC 751 CAGAGTGAAG CTCATGGCCT GCCCCTGCAC CTGCCTGGGA ACAAGAGCCC 801 CCACCGGGAC CCTGCCCCCC GGGGCCCTGC CCGCTTCCTG CCCCTGCCTG 851 GGCTGCCCCC AGCCCTGCCT GAGCCCCCAG GCATCCTGGC CCCCCAGCCC 901 CCAGATGTGG GCAGCAGTGA CCCCCTGTCC ATGGTGGGCG GCAGCCAGGG 951 CCGCAGCCCC AGCTATGAAA GT Coding sequence of SP1 - anti - albumin VHH - linker2 - FGF21 variant 2 SEQ ID NO: 43 1 ATGGGCTGGA GCTGCATCAT CCTCTTCCTG GTGGCCACGG CCACTGGAGT 51 TCATTCAGAA GTTCAGCTGG TGGAAAGTGG TGGTGGCCTG GTGCAGCCTG 101 GGAACAGCCT GCGCCTCAGC TGTGCTGCCT CGGGCTTCAC CTTCAGCAGC 151 TTTGGAATGT CCTGGGTGCG GCAGGCCCCG GGGAAAGGCC TGGAGTGGGT 201 CAGCAGCATT TCAGGAAGTG GCAGTGACAC CCTCTATGCA GATTCTGTAA 251 AAGGCCGCTT CACCATCAGC CGGGACAATG CCAAGACCAC CCTCTACCTG 301 CAGATGAACA GCCTGCGGCC GGAGGACACA GCTGTCTACT ACTGCACCAT 351 TGGTGGCAGC CTCAGCCGCA GCAGCCAGGG CACCCTGGTG ACTGTTTCTT 401 CAGGAGGTGG TGGCAGTGGT GGTGGCAGCC ACCCCATCCC AGACAGCAGC 451 CCCCTGCTGC AGTTTGGTGG CCAGGTGCGG CAGCGCTACC TCTACACAGA 501 TGATGCCCAG CAGACAGAAG CACATTTAGA AATAAGAGAA GATGGAACTG 551 TTGGTGGGGC GGCGGACCAG AGCCCTGAGA GCCTGCTGCA GCTGAAGGCC 601 TTGAAACCTG GTGTCATCCA GATTCTTGGA GTAAAAACCA GCCGCTTCCT 651 CTGCCAGAGG CCAGATGGGG CCCTCTATGG CAGCCTTCAT TTTGATCCTG 701 AGGCCTGCAG CTTCCGGGAG CGGCTGCTGG AAGATGGCTA CAATGTTTAT 751 CAAAGTGAAG CACATGGCCT GCCCCTGCAC CTTCCTGGAA ATAAAAGCCC 801 CCACCGGGAC CCGGCGCCCC GGGGCCCGGC CCGCTTCCTG CCCCTGCCTG 851 GGCTGCCCCC GGCGCTGCCT GAGCCGCCGG GCATCCTGGC CCCCCAGCCC 901 CCAGATGTGG GCAGCAGTGA CCCCCTCTCC ATGGTTGGTG GCAGCCAGGG 951 CCGCAGCCCC AGCTATGAAA GT Coding sequence of SP1 - anti - albumin VHH - linker 2 - FGF21 variant 2 SEQ ID NO: 44 1 ATGGGCTGGA GCTGCATCAT CCTCTTCCTG GTGGCCACGG CCACTGGAGT 51 TCATTCAGAA GTTCAGCTGG TGGAGAGTGG TGGTGGCCTG GTGCAGCCTG 101 GGAACAGCCT GCGCCTCAGC TGCGCGGCGT CGGGCTTCAC CTTCAGCAGC 151 TTTGGAATGT CCTGGGTGCG GCAGGCGCCG GGGAAGGGCC TGGAGTGGGT 201 CAGCAGCATC AGTGGAAGTG GCAGTGACAC CCTCTATGCA GATTCTGTGA 251 AGGGCCGCTT CACCATCAGC CGGGACAATG CCAAGACCAC CCTCTACCTG 301 CAGATGAACA GCCTGCGGCC GGAGGACACA GCTGTCTACT ACTGCACCAT 351 TGGTGGCAGC CTCAGCCGCA GCAGCCAGGG CACCCTGGTC ACTGTCAGCA 401 GTGGTGGTGG TGGCAGTGGT GGTGGCAGCC ACCCCATCCC AGACAGCAGC 451 CCCCTGCTGC AGTTTGGTGG CCAGGTGCGG CAGCGCTACC TCTACACAGA 501 TGATGCCCAG CAGACGGAGG CCCACCTGGA GATCCGAGAA GATGGAACTG 551 TTGGTGGCGC GGCGGACCAG AGCCCTGAGA GCCTGCTGCA GCTGAAGGCC 601 CTGAAGCCAG GTGTCATCCA GATTCTTGGT GTCAAGACCA GCCGCTTCCT 651 CTGCCAGAGG CCAGATGGGG CCCTCTATGG CAGCCTTCAT TTTGATCCTG 701 AGGCCTGCTC CTTCCGGGAG CGGCTGCTGG AGGATGGCTA CAATGTCTAC 751 CAGAGTGAAG CTCATGGACT TCCTCTTCAT CTTCCTGGGA ACAAGAGCCC 801 CCACCGGGAC CCGGCGCCGC GGGGCCCGGC GCGCTTCCTG CCCCTGCCTG 851 GGCTGCCGCC GGCGCTGCCT GAGCCGCCGG GCATCCTGGC GCCCCAGCCC 901 CCAGATGTGG GCAGCAGTGA CCCCCTCTCC ATGGTTGGTG GCAGCCAGGG 951 CCGCAGCCCC AGCTATGAAA GT Coding sequence of SP1 - anti - albumin VHH - linker 2 - FGF21 variant 2 SEQ ID NO: 45 1 ATGGGCTGGA GCTGCATCAT CCTCTTCCTG GTGGCCACGG CCACTGGAGT 51 TCATTCAGAG GTGCAGCTGG TGGAGAGTGG TGGTGGCCTG GTGCAGCCTG 101 GGAACAGCCT GCGCCTCAGC TGTGCTGCCT CGGGCTTCAC CTTCAGCAGC 151 TTTGGAATGT CCTGGGTGCG GCAGGCCCCG GGGAAGGGCC TGGAGTGGGT 201 CAGCAGCATC AGTGGCAGTG GCAGTGACAC CCTCTATGCA GATTCTGTGA 251 AGGGCCGCTT CACCATCAGC CGGGACAATG CCAAGACCAC CCTCTACCTG 301 CAGATGAACA GCCTGCGGCC GGAGGACACA GCTGTCTACT ACTGCACCAT 351 TGGTGGCAGC CTCAGCCGCA GCAGCCAGGG CACCCTGGTG ACTGTCAGCA 401 GTGGTGGTGG TGGCAGTGGT GGTGGCAGCC ACCCCATCCC AGACAGCAGC 451 CCCCTGCTGC AGTTTGGTGG CCAGGTGCGG CAGCGCTACC TCTACACAGA 501 TGATGCCCAG CAGACAGAGG CCCACCTGGA GATCAGAGAA GATGGAACAG 551 TTGGTGGTGC TGCTGACCAG AGCCCTGAGA GCCTGCTGCA GCTGAAGGCC 601 CTGAAGCCAG GTGTCATCCA GATCCTGGGT GTGAAGCCA GCCGCTTCCT 651 CTGCCAGAGG CCAGATGGGG CCCTCTATGG CAGCCTGCAC TTTGATCCTG 701 AGGCCTGCAG CTTCCGGGAG CGGCTGCTGG AGGATGGCTA CAATGTCTAC 751 CAGAGTGAAG CTCATGGCCT GCCCCTGCAC CTGCCTGGGA ACAAGAGCCC 801 CCACCGGGAC CCGGCGCCCC GGGGCCCGGC GCGCTTCCTG CCCCTGCCTG 851 GGCTGCCGCC GGCGCTGCCT GAGCCGCCGG GCATCCTGGC CCCCCAGCCCC 901 CCAGATGTGG GCAGCAGTGA CCCCCTCTCC ATGGTGGGCG GCAGCCAGGG 951 CCGCAGCCCC AGCTATGAAA GT SP1 - VHH regulator - type 2 - FGF21 is a 2-dimensional compound SEQ ID NO: 46 1 ATGGGCTGGA GCTGCATCAT CCTGTTCCTG GTGGCCACCG CCACCGGCGT 51 GCACAGCGAG GTGCAGCTGG TGGAGAGCGG CGGCGGCCTG GTGCAGCCTG 101 GCAACAGCCT GAGACTGAGC TGCGCCGCCA GCGGCTTCAC CTTCAGCAGC 151 TTCGGCATGA GCTGGGTGAG ACAGGCCCCT GGCAAGGGCC TGGAGTGGGT 201 GAGCAGCATC AGCGGCAGCG GCAGCGACAC CCTGTACGCC GACAGCGTGA 251 AGGGCAGATT CACCATCAGC AGAGACAACG CCAAGACCAC CCTGTACCTG 301 CAGATGAACA GCCTGAGACC TGAGGACACC GCCGTGTACT ACTGCACCAT 351 CGGCGGCAGC CTGAGCAGAA GCAGCCAGGG CACCCTGGTG ACCGTGAGCA 401 GCGGCGGCGG CGGCAGCGGC GGCGGCAGCC ACCCTATCCC TGACAGCAGC 451 CCTCTGCTGC AGTTCGGCGG CCAGGTGAGA CAGAGATACC TGTACACCGA 501 CGACGCCCAG CAGACCGAGG CCCACCTGGA GATCAGAGAG GACGGCACCG 551 TGGGCGGCGC CGCCGACCAG AGCCCTGAGA GCCTGCTGCA GCTGAAGGCC 601 CTGAAGCCTG GCGTGATCCA GATCCTGGGC GTGAAGACCA GCAGATTCCT 651 GTGCCAGAGA CCTGACGGCG CCCTGTACGG CAGCCTGCAC TTCGACCCTG 701 AGGCCTGCAG CTTCAGAGAG AGACTGCTGG AGGACGGCTA CAACGTGTAC 751 CAGAGCGAGG CCCACGGCCT GCCTCTGCAC CTGCCTGGCA ACAAGAGCCC 801 TCACAGAGAC CCTGCCCCTA GAGGCCCTGC CAGATTCCTG CCTCTGCCTG 851 GCCTGCCTCC TGCCCTGCCT GAGCCTCCTG GCATCCTGGC CCCTCAGCCT 901 CCTGACGTGG GCAGCAGCGA CCCTCTGAGC ATGGTGGGCG GCAGCCAGGG 951 CAGAAGCCCT AGCTACGAGA GC Coding sequence of SP1 - anti - albumin VHH - linker2 - FGF21 variant 2 SEQ ID NO: 47 1 ATGGGCTGGA GCTGCATCAT CCTCTTCCTG GTGGCCACAG CCACTGGAGT 51 TCATTCAGAA GTTCAGCTGG TGGAAAGTGG TGGTGGCCTG GTGCAGCCTG 101 GGAACAGCCT GCGCCTCAGC TGTGCTGCCA GTGGCTTCAC CTTCAGCAGC 151 TTTGGAATGT CCTGGGTGCG GCAGGCCCCT GGGAAAGGCC TGGAGTGGGT 201 CAGCAGCATC AGTGGCAGTG GCAGTGACAC CCTCTATGCA GATTCTGTGA 251 AGGGCCGCTT CACCATCAGC CGGGACAATG CCAAGACCAC CCTCTACCTG 301 CAGATGAACA GCCTGCGGCC GGAGGACACA GCTGTGTACT ACTGCACCAT 351 TGGTGGCAGC CTCAGCCGCA GCAGCCAGGG CACCCTGGTG ACTGTTTCTT 401 CAGGAGGTGG TGGCAGTGGT GGTGGCAGCC ACCCCATCCC AGACAGCAGC 451 CCCCTGCTGC AGTTTGGTGG CCAGGTGCGG CAGCGCTACC TCTACACAGA 501 TGATGCCCAG CAGACAGAAG CCCACCTGGA AATAAGAGAA GATGGAACTG 551 TTGGTGGTGC TGCTGACCAG AGCCCTGAGA GCCTGCTGCA GCTGAAGGCC 601 CTGAAGCCAG GTGTCATCCA GATTCTTGGT GTGAAGACCA GCCGCTTCCT 651 CTGCCAGAGG CCAGATGGGG CCCTCTATGG CAGCCTTCAT TTTGATCCTG 701 AGGCCTGCAG CTTCCGGGAG CGGCTGCTGG AAGATGGCTA CAATGTTTAT 751 CAAAGTGAAG CACATGGCCT GCCCCTGCAC CTGCCTGGAA ATAAAAGCCC 801 CCACAGAGAT CCTGCCCCCC GGGGCCCTGC CCGCTTCCTG CCCCTGCCTG 851 GGCTGCCCCC GGCCCTGCCT GAGCCGCCGG GCATCCTGGC CCCCCAGCCC 901 CCAGATGTGG GCAGCAGTGA CCCCCTCTCC ATGGTTGGTG GCAGCCAGGG 951 CCGCAGCCCC AGCTATGAAA GT Coding sequence of SP3 - GLP-1 – linker 3 - ABD - linker 2 - FGF21 variant 2 SEQ ID NO: 48 1 ATGGAGACCG ACACCCTGCT GCTGTGGGTG CTGCTGCTGT GGGTGCCTGG 51 CAGCACCGGC CACGGCGAGG GCACCTTCAC CAGCGACGTG AGCAGCTACC 101 TGGAGGAGCA GGCCGCCAAG GAGTTCATCG CCTGGCTGGT GAAGGGCGGC 151 GGCGGCGGCG GCGGCAGCGC CGAGGCCAAG GAGGCCGCCA ACGCCGAGCT 201 GGACAGCTAC GGCGTGAGCG ACTTCTACAA GAGACTGATC GACAAGGCCA 251 AGACCGTGGA GGGCGTGGAG GCCCTGAAGG ACGCCATCCT GGCCGCCCTG 301 CCTGGCGGCG GCGGCAGCGG CGGCGGCAGC CACCCTATCC CTGACAGCAG 351 CCCTCTGCTG CAGTTCGGCG GCCAGGTGAG ACAGAGATAC CTGTACACCG 401 ACGACGCCCA GCAGACCGAG GCCCACCTGG AGATCAGAGA GGACGGCACC 451 GTGGGCGGCG CCGCCGACCA GAGCCCTGAG AGCCTGCTGC AGCTGAAGGC 501 CCTGAAGCCT GGCGTGATCC AGATCCTGGG CGTGAAGACC AGCAGATTCC 551 TGTGCCAGAG ACCTGACGGC GCCCTGTACG GCAGCCTGCA CTTCGACCCT 601 GAGGCCTGCA GCTTCAGAGA GAGACTGCTG GAGGACGGCT ACAACGTGTA 651 CCAGAGCGAG GCCCACGGCC TGCCTCTGCA CCTGCCTGGC AACAAGAGCC 701 CTCACAGAGA CCCTGCCCCT AGAGGCCCTG CCAGATTCCT GCCTCTGCCT 751 GGCCTGCCTC CTGCCCTGCC TGAGCCTCCT GGCATCCTGG CCCCTCAGCC 801 TCCTGACGTG GGCAGCAGCG ACCCTCTGAG CATGGTGGGC GGCAGCCAGG 851 GCAGAAGCCC TAGCTACGAG AGC Coding sequence of SP3 - GLP-1 – linker 3 - ABD - linker 2 - FGF21 variant 2 SEQ ID NO: 49 1 ATGGAAACAG ACACGCTGCT GCTCTGGGTG CTGCTGCTCT GGGTTCCTGG 51 CAGCACTGGA CATGGAGAAG GCACCTTCAC CAGTGATGTC AGCAGCTACC 101 TGGAGGAGCA GGCGGCCAAG GAGTTCATTG CCTGGCTGGT GAAAGGTGGT 151 GGTGGTGGTG GTGGCTCGGC GGAGGCCAAG GAGGCGGCCA ATGCAGAGCT ' 201 GGACAGCTAT GGTGTCAGTG ACTTCTACAA GCGCCTCATT GACAAGGCCA 251 AGACAGTAGA AGGTGTGGAG GCCTTGAAAG ATGCCATCCT GGCGGCGCTG 301 CCTGGTGGTG GTGGCAGTGG TGGTGGCAGC CACCCCATCC CAGACAGCAG 351 CCCCCTGCTG CAGTTTGGTG GCCAGGTGCG GCAGCGCTAC CTCTACACAG 401 ATGATGCCCA GCAGACAGAG GCCCACCTGG AGATCAGAGA AGATGGAACT 451 GTTGGTGGCG CGGCGGACCA GAGCCCTGAG AGCCTGCTGC AGCTGAAGGC 501 CCTGAAGCCA GGTGTCATCC AGATCCTGGG GGTCAAGACC AGCCGCTTCC 551 TCTGCCAGAG GCCAGATGGG GCCCTCTATG GCAGCCTGCA CTTTGATCCT 601 GAGGCCTGCT CCTTCCGGGA GCGGCTGCTG GAGGATGGCT ACAATGTCTA 651 CCAGAGTGAA GCTCATGGCC TGCCCCTGCA CCTTCCTGGG AACAAGAGCC 701 CCCACCGGGA CCCGGCGCCG CGGGGCCCGG CGCGCTTCCT GCCCCTGCCT 751 GGGCTGCCGC CGGCGCTGCC TGAGCCGCCG GGCATCCTGG CGCCCCAGCC 801 CCCAGATGTG GGCAGCAGTG ACCCCCTCTC CATGGTTGGT GGCAGCCAGG 851 GCCGCAGCCC CAGCTATGAA AGT Coding sequence of SP3 - GLP-1 – linker 3 - ABD - linker 2 - FGF21 variant 2 SEQ ID NO: 50 1 ATGGAAACAG ACACCCTGCT GCTGTGGGTG CTGCTGCTGT GGGTTCCTGG 51 CAGCACTGGA CATGGAGAAG GCACCTTCAC CAGTGATGTC AGCAGCTACC 101 TGGAGGAGCA GGCGGCCAAA GAATTTATTG CCTGGCTGGT GAAAGGAGGT 151 GGTGGTGGTG GTGGCTCTGC AGAAGCAAAA GAAGCAGCAA ATGCAGAGCT 201 GGACAGCTAT GGTGTTTCTG ATTTTTATAA AAGATTAATT GATAAAGCAA 251 AAACTGTAGA AGGTGTGGAG GCCTTGAAAG ATGCCATCCT GGCGGCGCTG 301 CCTGGTGGTG GTGGCAGTGG TGGTGGCAGC CACCCCATCC CAGACAGCAG 351 CCCCCTGCTG CAGTTTGGTG GCCAGGTGCG GCAGCGCTAC CTCTACACAG 401 ATGATGCCCA GCAGACAGAA GCACATTTAG AAATAAGAGA AGATGGAACT 451 GTTGGTGGTG CTGCTGACCA GAGCCCTGAG AGCCTGCTGC AGCTGAAGGC 501 CTTGAAACCT GGTGTCATCC AGATTCTTGG AGTAAAAACC AGCCGCTTCC 551 TCTGCCAGAG GCCAGATGGG GCCCTCTATG GCAGCCTTCA TTTTGATCCT 601 GAGGCCTGCA GCTTCCGGGA GCGGCTGCTG GAAGATGGCT ACAATGTTTA 651 TCAAAGTGAA GCACATGGCC TGCCCCTGCA CCTTCCTGGA AATAAAAGCC 701 CCCACAGAGA TCCTGCCCCC CGGGGCCCTG CCCGCTTCCT GCCCCTGCCT 751 GGGCTGCCCC CGGCCCTGCC TGAGCCGCCG GGCATCCTGG CCCCCCAGCC 801 CCCAGATGTG GGCAGCAGTG ACCCCCTCTC CATGGTTGGT GGCAGCCAGG 851 GCCGCAGCCC CAGCTATGAA AGT Coding sequence of SP3 - GLP-1 – linker 3 - ABD - linker 2 - FGF21 variant 2 SEQ ID NO: 51 1 ATGGAAACAG ACACCCTGCT GCTGTGGGTG CTGCTGCTGT GGGTGCCTGG 51 CAGCACTGGA CATGGAGAGG GCACCTTCAC CAGTGATGTC AGCAGCTACC 101 TGGAGGAGCA GGCTGCCAAG GAGTTCATTG CCTGGCTGGT GAAAGGAGGA 151 GGAGGAGGAG GAGGCTCTGC AGAGGCCAAG GAGGCTGCCA ATGCAGAGCT 201 GGACAGCTAT GGTGTCAGTG ACTTCTACAA GCGCCTCATT GACAAGGCCA 251 AGACAGTGGA GGGTGTGGAG GCCCTGAAGG ATGCCATCCT GGCGGCGCTG 301 CCTGGAGGAG GAGGCAGTGG TGGTGGCAGC CACCCCATCC CAGACAGCAG 351 CCCCCTGCTG CAGTTTGGGG GCCAGGTGCG GCAGCGCTAC CTCTACACAG 401 ATGATGCCCA GCAGACAGAG GCCCACCTGG AGATCAGAGA AGATGGAACA 451 GTTGGTGGTG CTGCTGACCA GAGCCCTGAG AGCCTGCTGC AGCTGAAGGC 501 CCTGAAGCCT GGGGTCATCC AGATCCTGGG CGTGAAGACC AGCCGCTTCC 551 TCTGCCAGAG GCCAGATGGG GCCCTGTATG GCAGCCTGCA CTTTGATCCT 601 GAGGCCTGCT CCTTCCGGGA GCGGCTGCTG GAGGATGGCT ACAATGTCTA 651 CCAGAGTGAA GCTCATGGCC TGCCCCTGCA CCTGCCTGGG AACAAGAGCC 701 CCCACCGGGA CCCTGCCCCC CGGGGCCCTG CCCGCTTCCT GCCCCTGCCT 751 GGGCTGCCCC CAGCCCTGCC TGAGCCCCCA GGCATCCTGG CCCCCCAGCC 801 CCCAGATGTG GGCAGCAGTG ACCCCCTGTC CATGGTGGGC GGCAGCCAGG 851 GCCGCAGCCC CAGCTATGAA AGT Coding sequence of SP3 - GLP-1 – linker 3 - ABD - linker 2 - FGF21 variant 2 SEQ ID NO: 52 1 ATGGAAACAG ACACCCTGCT GCTGTGGGTG CTGCTGCTGT GGGTGCCTGG 51 CAGCACTGGA CATGGAGAGG GCACCTTCAC CAGTGATGTC AGCAGCTACC 101 TGGAGGAGCA GGCTGCCAAG GAGTTCATTG CCTGGCTGGT GAAAGGTGGT 151 GGTGGTGGTG GTGGCTCTGC AGAGGCCAAG GAGGCGGCCA ATGCAGAGCT 201 GGACAGCTAT GGTGTCAGTG ACTTCTACAA GCGCCTCATT GACAAGGCCA 251 AGACAGTAGA AGGTGTGGAG GCCCTGAAGG ATGCCATCCT GGCGGCGCTG 301 CCTGGTGGTG GTGGCAGTGG TGGTGGCAGC CACCCCATCC CAGACAGCAG 351 CCCCCTGCTG CAGTTTGGTG GCCAGGTGCG GCAGCGCTAC CTCTACACAG 401 ATGATGCCCA GCAGACAGAG GCCCACCTGG AGATCAGAGA AGATGGAACT 451 GTTGGTGGTG CTGCTGACCA GAGCCCTGAG AGCCTGCTGC AGCTGAAGGC 501 CCTGAAGCCA GGTGTCATCC AGATCCTGGG CGTGAAGACC AGCCGCTTCC 551 TCTGCCAGAG GCCAGATGGG GCCCTCTATG GCAGCCTGCA CTTTGATCCT 601 GAGGCCTGCT CCTTCCGGGA GCGGCTGCTG GAGGATGGCT ACAATGTCTA 651 CCAGAGTGAA GCTCATGGCC TGCCCCTGCA CCTGCCTGGG AACAAGAGCC 701 CCCACCGGGA CCCGGCGCCC CGGGGCCCTG CCCGCTTCCT GCCCCTGCCT 751 GGGCTGCCGC CGGCGCTGCC TGAGCCGCCG GGCATCCTGG CCCCCCAGCC 801 CCCAGATGTG GGCAGCAGTG ACCCCCTCTC CATGGTGGGC GGCAGCCAGG 851 GCCGCAGCCC CAGCTATGAA AGT Coding sequence of SP3 - GLP - 1 - linker 3 - ABD - linker 2 - FGF21 variant 2 SEQ ID NO: 53 1 ATGGAAACAG ACACCCTGCT GCTGTGGGTG CTGCTGCTGT GGGTGCCTGG 51 CAGCACTGGA CATGGAGAAG GCACCTTCAC CAGTGATGTC AGCAGCTACC 101 TGGAGGAGCA GGCTGCCAAG GAATTTATTG CCTGGCTGGT GAAAGGAGGA 151 GGAGGAGGAG GAGGCTCTGC AGAGGCCAAG GAAGCTGCCA ATGCAGAGCT 201 GGACAGCTAT GGAGTTTCTG ACTTCTACAA GCGCCTCATT GATAAAGCCA 251 AGACTGTGGA AGGTGTGGAG GCCCTGAAAG ATGCCATCCT GGCTGCCCTG 301 CCTGGAGGAG GAGGAAGTGG TGGTGGCAGC CACCCCATCC CAGACAGCAG 351 CCCCCTGCTG CAGTTTGGGG GCCAGGTGCG GCAGCGCTAC CTCTACACAG 401 ATGATGCCCA GCAGACAGAA GCCCACCTGG AGATCAGAGA AGATGGAACT 451 GTTGGAGGAG CTGCTGACCA GAGCCCTGAG AGCCTGCTGC AGCTGAAGGC 501 CCTGAAGCCT GGGGTCATCC AGATCCTGGG GGTGAAGACC AGCCGCTTCC 551 TCTGCCAGAG GCCAGATGGG GCCCTGTATG GCAGCCTGCA CTTTGATCCT 601 GAGGCCTGCA GCTTCCGGGA GCGGCTGCTG GAAGATGGCT ACAATGTCTA 651 CCAGAGTGAA GCTCATGGCC TGCCCCTGCA CCTGCCTGGA AATAAAAGCC 701 CCCACAGAGA TCCTGCCCCC CGGGGCCCTG CCCGCTTCCT GCCCCTGCCT 751 GGGCTGCCCC CAGCCCTGCC TGAGCCCCCA GGCATCCTGG CCCCCCAGCC 801 CCCAGATGTG GGCAGCAGTG ACCCCCTGTC CATGGTGGGC GGCAGCCAGG 851 GCCGCAGCCC CAGCTATGAA AGT Coding sequence of SP3 - GLP - 1 - linker 2 - anti - albumin VHH - linker 2 - FGF21 variant 2 SEQ ID NO: 54 1 ATGGAGACAG ACACTCTGCT GCTGTGGGTG CTGCTCCTCT GGGTTCCTGG 51 CAGCACTGGA CATGGAGAGG GCACCTTCAC TTCAGATGTC TCCTCCTACT 101 TGGAGGAGCA GGCTGCCAAG GAGTTCATCG CCTGGCTGGT CAAGGGCGGC 151 GGGGGAGGTG GAGGCTCAGG AGGGGGCTCT GAGGTGCAGC TGGTGGAGAG 201 CGGCGGTGGC CTCGTCCAGC CTGGAAATTC TCTGCGGCTG AGCTGTGCCG 251 CCAGTGGCTT CACATTTTCT TCCTTTGGCA TGTCCTGGGT GCGCCAGGCC 301 CCAGGAAAAG GCCTGGAGTG GGTCAGCAGC ATCTCTGGAA GTGGATCTGA 351 CACCCTGTAT GCAGATTCTG TGAAAGGAAG GTTCACCATC AGCCGGGACA 401 ACGCCAAAAC CACCCTCTAC CTCCAGATGA ACAGCCTGAG GCCTGAGGAC 451 ACAGCTGTCT ACTACTGCAC CATTGGAGGA AGCCTCAGCC GCTCCAGCCA 501 GGGGACCTTG GTGACTGTGT CCTCTGGGGG TGGGGGCAGT GGTGGTGGCA 551 GCCACCCCAT CCCAGACAGC TCCCCACTGC TTCAGTTTGG AGGCCAGGTG 601 CGGCAGCGCT ACCTGTACAC AGATGATGCC CAGCAGACAG AGGCCCACCT 651 GGAGATCCGG GAGGATGGAA CAGTGGGAGG AGCTGCTGAC CAGAGCCCAG 701 AAAGCCTTCT GCAGCTCAAG GCCCTGAAGC CAGGGGTCAT CCAGATCCTC 751 GGTGTGAAGA CCAGCAGATT TCTGTGCCAG AGGCCAGATG GGGCCCTCTA 801 TGGATCCCTG CATTTTGATC CTGAAGCCTG CAGCTTCAGA GAAAGACTCC 851 TGGAAGATGG CTACAACGTG TACCAGAGTG AAGCTCATGG CCTTCCTCTG 901 CACCTCCCCG GCAACAAGAG CCCCCACAGG GACCCTGCCC CTCGGGGCCC 951 AGCCCGCTTC CTGCCCCTCC CTGGGCTGCC TCCTGCTCTG CCAGAGCCCC 1001 CAGGCATCCT GGCCCCCCAG CCCCCTGATG TGGGGAGCAG TGACCCCCTG 1051 TCCATGGTGG GCGGCTCCCA GGGCCGCAGC CCTTCCTATG AAAGT Coding sequence of SP3 - GLP - 1 - linker 2 - anti - albumin VHH - linker 2 - FGF21 variant 2 SEQ ID NO: 55 1 ATGGAGACCG ACACCCTGCT GCTGTGGGTG CTGCTGCTGT GGGTGCCTGG 51 CAGCACCGGC CACGGCGAGG GCACCTTCAC CAGCGACGTG AGCAGCTACC 101 TGGAGGAGCA GGCCGCCAAG GAGTTCATCG CCTGGCTGGT GAAGGGCGGC 151 GGCGGCGGCG GCGGCAGCGG CGGCGGCAGC GAGGTGCAGC TGGTGGAGAG 201 CGGCGGCGGC CTGGTGCAGC CTGGCAACAG CCTGAGACTG AGCTGCGCCG 251 CCAGCGGCTT CACCTTCAGC AGCTTCGGCA TGAGCTGGGT GAGACAGGCC 301 CCTGGCAAGG GCCTGGAGTG GGTGAGCAGC ATCAGCGGCA GCGGCAGCGA 351 CACCCTGTAC GCCGACAGCG TGAAGGGCAG ATTCACCATC AGCAGAGACA 401 ACGCCAAGAC CACCCTGTAC CTGCAGATGA ACAGCCTGAG ACCTGAGGAC 451 ACCGCCGTGT ACTACTGCAC CATCGGCGGC AGCCTGAGCA GAAGCAGCCA 501 GGGCACCCTG GTGACCGTGA GCAGCGGCGG CGGCGGCAGC GGCGGCGGCA 551 GCCACCCTAT CCCTGACAGC AGCCCTCTGC TGCAGTTCGG CGGCCAGGTG 601 AGACAGAGAT ACCTGTACAC CGACGACGCC CAGCAGACCG AGGCCCACCT 651 GGAGATCAGA GAGGACGGCA CCGTGGGCGG CGCCGCCGAC CAGAGCCCTG 701 AGAGCCTGCT GCAGCTGAAG GCCCTGAAGC CTGGCGTGAT CCAGATCCTG 751 GGCGTGAAGA CCAGCAGATT CCTGTGCCAG AGACCTGACG GCGCCCTGTA 801 CGGCAGCCTG CACTTCGACC CTGAGGCCTG CAGCTTCAGA GAGAGACTGC 851 TGGAGGACGG CTACAACGTG TACCAGAGCG AGGCCCACGG CCTGCCTCTG 901 CACCTGCCTG GCAACAAGAG CCCTCACAGA GACCCTGCCC CTAGAGGCCC 951 TGCCAGATTC CTGCCTCTGC CTGGCCTGCC TCCTGCCCTG CCTGAGCCTC 1001 CTGGCATCCT GGCCCCTCAG CCTCCTGACG TGGGCAGCAG CGACCCTCTG 1051 AGCATGGTGG GCGGCAGCCA GGGCAGAAGC CCTAGCTACG AGAGC Coding sequence of SP3 - GLP-1 - linker 2 - anti-albumin VHH - linker 2 - FGF21 variant 2 SEQ ID NO: 56 1 ATGGAGACAG ACACCCTCCT GCTCTGGGTG CTGCTGCTGT GGGTTCCTGG 51 CAGCACTGGA CATGGAGAGG GCACCTTCAC TTCAGATGTC AGCAGCTACT 101 TGGAGGAGCA GGCTGCCAAG GAGTTCATCG CCTGGCTGGT CAAGGGCGGG 151 GGTGGTGGAG GTGGCAGTGG TGGGGGCTCT GAGGTGCAGC TGGTGGAGAG 201 CGGCGGTGGC CTCGTCCAGC CTGGAAATTC TCTGCGGCTG AGCTGTGCCG 251 CCAGTGGCTT CACATTTTCT TCCTTTGGCA TGTCCTGGGT GCGGCAGGCC 301 CCAGGAAAAG GCCTGGAGTG GGTCTCCAGC ATCTCTGGAA GTGGATCTGA 351 CACTCTGTAT GCAGATTCTG TGAAAGGAAG GTTCACCATC AGCCGGGACA 401 ACGCCAAAAC CACCCTGTAC CTGCAGATGA ACAGCCTGAG GCCTGAGGAC 451 ACAGCTGTCT ACTACTGCAC CATTGGAGGC TCCCTCAGCC GCTCCTCCCA 501 GGGGACCTTG GTGACTGTGT CCTCTGGGGG CGGCGGCTCA GGAGGAGGGA 551 GCCACCCCAT CCCAGACAGC TCCCCACTGC TTCAGTTTGG AGGCCAGGTG 601 CGCCAGCGCT ACCTCTACAC AGATGATGCC CAGCAGACAG AGGCCCACCT 651 GGAGATCCGG GAGGATGGAA CAGTGGGGGG AGCTGCTGAC CAGAGCCCAG 701 AAAGCCTCCT CCAGCTCAAG GCCCTGAAGC CAGGGGTCAT CCAGATCCTC 751 GGTGTGAAGA CCAGCAGATT TCTGTGCCAG AGGCCAGATG GGGCCCTCTA 801 TGGATCCCTG CATTTTGATC CTGAAGCCTG CAGCTTCAGA GAAAGACTTC 851 TGGAAGATGG CTACAACGTG TACCAGAGTG AAGCTCATGG CCTTCCTCTG 901 CACCTCCCCG GCAACAAGAG CCCCCACAGG GACCCTGCCC CTCGGGGCCC 951 AGCCCGCTTC CTGCCCCTCC CTGGGCTGCC TCCTGCTCTG CCAGAGCCCC 1001 CAGGCATCCT GGCCCCCCAG CCCCCTGATG TGGGCAGCAG TGACCCCCTG 1051 TCCATGGTGG GAGGAAGCCA GGGCCGCAGC CCTTCCTATG AAAGT Coding sequence of SP3 - GLP - 1 - linker 2 - anti - albumin VHH - linker 2 - FGF21 variant 2 SEQ ID NO: 57 1 ATGGAGACAG ACACTCTGCT CCTCTGGGTG CTGCTGCTGT GGGTTCCTGG 51 GAGCACTGGA CATGGAGAGG GCACCTTCAC TTCAGATGTC TCCTCCTACT 101 TGGAGGAGCA GGCTGCCAAG GAGTTCATCG CCTGGCTGGT GAAAGGAGGT 151 GGGGGCGGTG GTGGCTCAGG AGGAGATCT GAGGTGCAGC TCGTGGAGAG 201 CGGCGGCGGG CTGGTCCAGC CTGGAAATTC TCTGCGGCTG AGCTGTGCCG 251 CCTCTGGCTT CACATTTTCT TCCTTTGGCA TGTCCTGGGT GCGCCAGGCC 301 CCAGGAAAAG GCCTGGAGTG GGTCAGCAGC ATCTCTGGAA GTGGCTCTGA 351 CACCCTCTAT GCAGATTCTG TCAAGGGCCG CTTCACCATC AGCCGGGACA 401 ACGCCAAAAC CACCCTGTAC CTGCAGATGA ACAGCCTGAG GCCTGAGGAC 451 ACAGCTGTCT ACTACTGCAC CATTGGAGGC AGCCTCAGCC GCTCCAGCCA 501 GGGGACCTTG GTGACTGTGT CCTCTGGGGG TGGCGGCAGT GGAGGGGAA 551 GCCACCCCAT CCCAGACAGC TCCCCACTGC TTCAGTTTGG AGGCCAGGTG 601 CGGCAGCGCT ACCTCTACAC BUYGCC CAGCAGACAG AGGCCCACCT 651 GGAGATCCGG GAGGGGA CAGTGGGTGG AGCTGCTGAC CAGAGCCCAG 701 AAAGCCTTCT CCAGCTGAAG GCCCTCAAGC CAGGGGTCAT CCAGATCCTC 751 GGTGTGAAGA CCAGCAGATT TCTGTGCCAG AGGCCAGATG GGGCCCTGTA 801 TGGATCCCTG CACTTTGATC CTGAAGCCTG CAGCTTCAGA GAAAGACTCC 851 TGGAAGATGG CTACAACGTG TACCAGAGTG AAGCTCATGG CCTCCCCCTC 901 CACCTCCCTG GCAACAAGAG CCCCCACAGG GACCCTGCCC CTCGGGGCCC 951 AGCCAGGTTC CTGCCTCTGC CCGGCCTTCC TCCTGCTCTG CCAGAGCCCC 1001 CAGGCATCCT GGCCCCCCAG CCCCCTGATG TGGGCAGCAG TGACCCCCTG 1051 TCCATGGTGG GGGGCTCCCA GGGCCGGAGC CCTTCCTATG AAAGT Coding sequence of SP3 - GLP - 1 - linker2 - anti - albumin VHH - linker2 - FGF21 variant 2 SEQ ID NO: 58 1 ATGGAGACAG ACACCCTCCT GCTGTGGGTG CTGCTGCTCT GGGTTCCAGG 51 GAGCACTGGA CATGGAGAGG GCACCTTCAC TTCAGATGTG TCCTCCTACT 101 TGGAGGAGCA GGCTGCCAAG GAGTTCATCG CCTGGCTGGT GAAAGGAGGG 151 GGCGGGGGTG GTGGCTCAGG AGGAGGATCT GAGGTGCAGC TCGTGGAGAG 201 TGGAGGTGGG CTGGTCCAGC CTGGAAATTC TCTGCGGCTG AGCTGTGCCG 251 CCTCTGGCTT CACATTTTCT TCCTTTGGCA TGTCCTGGGT GAGGCAGGCC 301 CCAGGAAAAG GCCTGGAGTG GGTCTCCAGC ATCTCTGGAA GTGGCTCTGA 351 CACTCTGTAT GCAGATTCTG TCAAGGGCCG CTTCACCATC AGCCGGGACA 401 ACGCCAAAAC AACCCTGTAC CTGCAGATGA ACAGCCTGAG GCCAGAGGAC 451 ACAGCTGTCT ACTACTGCAC CATTGGTGGC TCCCTCAGCC GCAGCTCCCA 501 GGGCACACTG GTTACTGTGT CTTCTGGGGG GGGCGGCAGT GGTGGAGGCA 551 GCCACCCCAT CCCAGACAGC AGCCCACTGC TTCAGTTTGG AGGCCAGGTG 601 CGGCAGAGAT ACCTCTACAC AGATGATGCC CAGCAGACAG AGGCCCACCT 651 GGAAATTCGG GAGGATGGAA CAGTGGGGGG AGCTGCTGAC CAGAGCCCTG 701 AGAGCCTCCT CCAGCTGAAG GCCCTGAAAC CTGGTGTCAT CCAGATCCTC 751 GGTGTGAAGA CCAGCAGATT TCTGTGCCAG AGGCCTGATG GGGCCCTCTA 801 TGGATCCCTG CATTTTGATC CTGAAGCCTG CAGCTTCAGA GAAAGACTGT 851 TGGAAGATGG CTACAACGTG TACCAGAGTG AAGCTCATGG CCTCCCTCTG 901 CACCTCCCAG GCAACAAGAG CCCCCACAGA GACCCTGCCC CTCGGGGCCC 951 AGCCAGGTTC CTGCCCCTGC CTGGGCTTCC TCCAGCTCTG CCAGAACCTC 1001 CTGGCATCCT GGCCCCCCAG CCCCCAGATG TTGGAAGCAG TGACCCCCTC 1051 TCCATGGTGG GCGGCAGCCA GGGAAGATCT CCATCCTATG AAAGT Coding sequence of SP3 - GLP - 1 - linker 2 - anti - albumin VHH - linker 2 - FGF21 variant 2 SEQ ID NO: 59 1 ATGGAAACAG ACACGCTGCT GCTCTGGGTG CTGCTGCTCT GGGTTCCTGG 51 CAGCACTGGA CATGGAGAAG GCACCTTCAC CAGTGATGTC AGCAGCTACC 101 TGGAGGAGCA GGCGGCCAAG GAGTTCATTG CCTGGCTGGT GAAAGGTGGT 151 GGTGGTGGTG GTGGCAGTGG TGGTGGCAGT GAAGTTCAGC TGGTGGAGAG 201 TGGTGGTGGC CTGGTGCAGC CTGGGAACAG CCTGCGCCTC AGCTGTGCTG 251 CCTCGGGCTT CACCTTCAGC AGCTTTGGAA TGTCCTGGGT GCGGCAGGCG 301 CCGGGGAAGG GCCTGGAGTG GGTCAGCAGC ATCAGTGGCA GTGGAAGTGA 351 CACCCTCTAT GCAGATTCTG TGAAGGGCCG CTTCACCATC AGCCGGGACA 401 ATGCCAAGAC CACCCTCTAC CTGCAGATGA ACAGCCTGCG GCCGGAGGAC 451 ACAGCTGTCT ACTACTGCAC CATTGGTGGC AGCCTCAGCC GCAGCAGCCA 501 GGGCACCCTG GTCACTGTCA GCAGTGGTGG TGGTGGCAGT GGTGGTGGCA 551 GCCACCCCAT CCCAGACAGC AGCCCCCTGC TGCAGTTTGG TGGCCAGGTG 601 CGGCAGCGCT ACCTCTACAC AGATGATGCC CAGCAGACAG AAGCTCATTT 651 AGAAATAAGA GAAGATGGAA CTGTTGGTGG GGCGGCGGAC CAGAGCCCTG 701 AGAGCCTGCT GCAGCTGAAG GCCCTGAAGC CAGGTGTCAT CCAGATCCTG 751 GGGGTCAAGA CCAGCCGCTT CCTCTGCCAG AGGCCAGATG GGGCCCTCTA 801 TGGCAGCCTT CATTTTGATC CTGAGGCCTG CTCCTTCCGG GAGCGGCTGC 851 TGGAGGATGG CTACAATGTC TACCAGAGTG AAGCTCATGG ACTTCCTCTT 901 CATCTTCCTG GGAACAAGAG CCCCCACCGG GACCCGGCGC CGCGGGGCCC 951 GGCGCGCTTC CTGCCCCTGC CTGGGCTGCC GCCGGCGCTG CCTGAGCCGC 1001 CGGGCATCCT GGCGCCCCAG CCCCCAGATG TGGGCAGCAG TGACCCCCTC 1051 TCCATGGTTG GTGGCAGCCA GGGCCGCAGC CCCAGCTATG AAAGT Coding sequence of SP3 - IgG1 Fc - linker1 - FGF21 variant 2 SEQ ID NO: 60 1 ATGGAAACAG ACACGCTGCT GCTCTGGGTG CTGCTGCTCT GGGTTCCTGG 51 CAGCACTGGA CATGGAGAG GCACCTTCAC CAGTGATGTC AGCAGCTATT 101 TAGAAGAACA AGCAGCAAAA GAATTTATTG CCTGGCTGGT GAAAGGTGGT 151 GGTGGTGGTG GTGGCAGTGG TGGTGGTGGC AGTGGTGGTG GTGGCTCTGA 201 TAAAACCCAC ACCTGCCCGC CCTGCCCGGC GCCGGAGCTG CTGGGGGGCC 251 CCAGTGTCTT CCTCTTCCCG CCCAAGCCCA AGGACACCCT CATGATTTCA 301 AGAACTCCAG AAGTCACCTG TGTGGTGGTG GATGTTTCTC ATGAAGATCC 351 TGAGGTGAAG TTCAACTGGT ATGTTGATGG TGTTGAAGTT CATAATGCCA 401 AGACCAAGCC CCGGGAGGAG CAGTACAACA GCACCTACAG AGTTGTTTCT 451 GTTTTAACTG TTCTTCATCA GGACTGGTTA AATGGAAAAG AAATAAAATG 501 TAAAGTTTCA AATAAAGCCC TGCCCGCGCC CATTGAAAAA ACCATCTCCA 551 AAGCAAAAGG ACAGCCCCGG GAGCCCCAGG TGTACACGCT GCCGCCCAGC 601 AGAGATGAGC TGACCAAAA TCAAGTCAGC CTCACCTGCC TGGTGAAGGG 651 CTTCTACCCC AGTGATATTG CTGTGGAGTG GGAGAGCAAT GGGCAGCCAG 701 AAAATAATTA TAAAACCACC CCGCCGGTGC TGGACAGTGA TGGCTCCTTC 751 TTCCTCTACA GCAAATTAAC TGTGGACAAG AGCCGCTGGC AGCAAGGAAA 801 TGTCTTCAGC TGCAGTGTCA TGCATGAAGC CCTGCACAAC CACTACACCC 851 AGAAAAGCCT CAGCCTCAGC CCTGGGAAAG GAGGTGGTGG CAGTGGTGGT 901 GGTGGCAGTG GTGGTGGTGG CAGCCACCCC ATCCCAGACA GCAGCCCCCT 951 GCTGCAGTTT GGTGGCCAGG TGCGGCAGCG CTACCTCTAC ACAGATGATG 1001 CCCAGCAGAC AGAAGCACAT TTAGAAATAA GAGAAGATGG AACTGTTGGT 1051 GGTGCTGCTG ACCAGAGCCC TGAGAGCCTG CTGCAGCTGA AGGCCCTGAA 1101 GCCAGGTGTC ATCCAGATTC TTGGTGTGAA GACCAGCCGC TTCCTCTGCC 1151 AGAGGCCAGA TGGGGCCCTC TATGGCAGCC TTCATTTTGA TCCTGAGGCC 1201 TGCAGCTTCC GGGAGCGGCT GCTGGAAGAT GGCTACAATG TTTATCAAAG 1251 TGAAGCACAT GGCCTGCCCC TGCACCTTCC TGGAAATAAA AGCCCCCACA 1301 GAGATCCTGC CCCCCGGGGC CCTGCCCGCT TCTTGCCCCT GCCTGGGCTG 1351 CCCCCGGCCC TGCCTGAGCC GCCGGGCATC CTGGCCCCCC AGCCCCCAGA 1401 TGTGGGCAGC AGTGACCCCC TCTCCATGGT TGGTGGCAGC CAGGGCCGCA 1451 GCCCCAGCTA TGAAAGT SP3 - GLP-1 - Binding1 - IgG4 Fc - Binding1 - FGF21 and 2 complexes SEQ ID NO: 61 1 ATGGAAACAG ACACGCTGCT GCTCTGGGTG CTGCTGCTCT GGGTTCCTGG 51 CAGCACTGGA CATGGAGAAG GCACCTTCAC CAGTGATGTC AGCAGCTATT 101 AGCAGCAAAA GAATTTATTG CCTGGCTGGT GAAAGGTGGT 151 GGTGGTGGTG GTGGCAGTGG TGGTGGTGGC AGTGGTGGTG GTGGCTCTGC 201 AGAAAGTAAA TATGGGCCGC CCTGCCCGCC CTGCCCGGCG CCGGAGGCGG 251 CGGGGGGCCC CAGTGTCTTC CTCTTCCCGC CCAAGCCCA GGACACCCTC 301 ATGATTTCAA GAACTCCAGA AGTCACCTGT GTGGTGGTGG ATGTTTCTCA 351 AGAAGATCCT GAAGTTCAGT TCAACTGGTA TGTAGATGGT GTTGAAGTTC 401 ATATGCCAA GACCAAGCCC CGGGAGGAGC AGTTCAACAG CACCTACAGA 451 GTTGTTTCTG TTTTAACTGT TTTACATCAA ATGGAAAAGA 501 ATATAAATGT AAAGTTTCAA ATAAAGGCCT GCCCAGCAGC ATTGAAAAAA 551 CCATCTCCAA AGCAAAAGGA CAGCCCCGGG AGCCCCAGGT CTACACGCTG 601 CCGCCCAGCC AAGAAGAAAT GACCAAAAAT CAAGTTTCTC TCACCTGCCT 651 GGTGAAGGGC TTCTACCCCA GTGATATTGC TGTGGAGTGG GAGAGCAATG 701 GGCAGCCAGA AAATAATTAT AAAACCACCC CGCCGGTGCT GGACAGTGAT 751 GGCTCCTTCT TCCTCTACAG CCGCCTCACT GTGGACAAGA GCCGCTGGCA 801 AGAAGGAAAT GTCTTCTCCT GCAGTGTCAT GCATGAAGCC CTGCACAACC 851 ACTACACCCA GAAAAGCCTC AGCCTCAGCC TGGGCGGAGG TGGTGGCAGT 901 GGTGGTGGTG GCAGTGGTGG TGGTGGCAGC CACCCCATCC CAGACAGCAG 951 CCCCCTGCTG CAGTTTGGTG GCCAGGTGCG GCAGCGCTAC CTCTACACAG 1001 ATGATGCCCA GCAGACAGAA GCACATTTAG AAATAAGAGA AGATGGAACT 1051 GTTGGTGGTG CTGCTGACCA GAGCCCTGAG AGCCTGCTGC AGCTGAAGGC 1101 CCTGAAGCCA GGTGTCATCC AGATTCTTGG TGTGAAGACC AGCCGCTTCC 1151 TCTGCCAGAG GCCAGATGGG GCCCTCTATG GCAGCCTTCA TTTTGATCCT 1201 GAGGCCTGCA GCTTCCGGGA GCGGCTGCTG GAAGATGGCT ACAATGTTTA 1251 TCAAAGTGAA GCACATGGCC TGCCCCTGCA CCTTCCTGGA AATAAAAGCC 1301 CCCACAGAGATCCTGCCCCC CGGGGCCCTG CCCGCTTCCT GCCCCTGCCT 1351 GGGCTGCCCC CGGCCCTGCC TGAGCCGCCG GGCATCCTGG CCCCCCAGCC 1401 CCCAGATGTG GGCAGCAGTG ACCCCCTCTC CATGGTTGGT GGCAGCCAGG 1451 GCCGCAGCCC CAGCTATGAA AGT Amino acid sequence of signal peptide 1 (SP1) SEQ ID NO: 62 MGWSCIILFLVATATGVHS Amino acid sequence of signal peptide 2 (SP2) SEQ ID NO: 63 METDTLLLMVLLLWVPGSTG Amino acid sequence of signal peptide 2 (SP3) SEQ ID NO: 64 METDTLLLWVLLLWVPGSTG Amino acid sequence of IgG 1 Fc SEQ ID NO: 65 DKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIE KTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK Amino acid sequence of IgG 1 Fc (YTE) SEQ ID NO: 66 DKTHTCPPCPAPELLGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIE KTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK Amino acid sequence of IgG 4 Fc SEQ ID NO: 67 AESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSS IEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG The amino acid sequence of the albumin-binding domain SEQ ID NO: 68 AEAKEAANAELDSYGVSDFYKRLIDKAKTVEGVEALKDAILAALP Amino acid sequence of VHH antibody against human albumin SEQ ID NO: 69 EVQLVESGGGLVQPGNSLRLSCAASGFTFSSFGMSWVRQAPGKGLEWVSSISGSGSDTLYADSVKGRFTISRDNAKTTLYLQMNSLRPEDTAVYYCTIGGSSLRSSQGTLVTVSS amino acid sequence of linker 1 SEQ ID NO: 70 GGGGSGGGGSGGGGS amino acid sequence of linker 2 SEQ ID NO: 71 GGGGSGGGS amino acid sequence of linker 3 SEQ ID NO: 72 GGGGS Amino acid sequence of linker 4 SEQ ID NO: 73 GGGGGSGGGSGGGGS Amino acid sequence of recombinant human FGF21 SEQ ID NO: 74 MDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKGGGGSGGGGSGGGGSHPIPDSSPLLQFGGQVRQRYLYTDDAQQTEAHLEIREDGTVGGAADQSPESLLQLKALKPGVIQILGVKTSRFLCQRPDGALYGSLHFDPEACSFRERLLEDGYNVYQSEAHGLPLELPGNKSPHRDPAPRGPARFLPLPGLPPAPPEPPGILAPQPPDVGSSDPLSMVGGSQGRSPSYES Coding sequence of FGF21 variant 2 SEQ ID NO: 75 1 CACCCCATCC CGGACAGCAG CCCGCTGCTG CAGTTTGGAG GACAAGTTCG 51 TCAGCGCTAC CTCTACACAG ATGATGCCCA GCAAACAGAA GCACATTTAG 101 AAATAAGAGA AGATGGAACT GTTGGTGGGG CGGCGGACCA AAGTCCTGAA 151 AGTTTATTAC AATTAAAAGC ATTAAAACCT GGTGTTATTC AAATATTAGG 201 AGTAAAAACC AGCCGCTTCC TCTGCCAAAG ACCAGATGGT GCTTTATATG 251 GAAGTTTACA TTTTGATCCT GAGGCCTGCT CCTTCAGAGA AAGATTATTA 301 GAAGATGGTT ATAATGTTTA TCAAAGTGAA GCACATGGTC TTCCTTTACA 351 TCTTCCTGGA AATAAATCAC CCCACCGGGA CCCGGCGCCG CGGGGCCCGG 401 CGCGCTTCCT TCCTCTTCCT GGGCTGCCGC CGGCGCTGCC GGAGCCGCCG 451 GGCATCCTGG CGCCGCAGCC GCCGGATGTT GGCAGCAGTG ACCCCCTCTC 501 CATGGTTGGT GGCAGCCAAG GCCGCTCGCC CAGCTATGAA AGT Coding sequence of FGF21 variant 2 SEQ ID NO: 76 1 CACCCCATCC CAGACAGCAG CCCCCTGCTG CAGTTTGGTG GCCAGGTGCG 51 GCAGCGCTAC CTCTACACAG ATGATGCCCA GCAGACAGAA GCACATTTAG 101 AAATAAGAGA AGATGGAACT GTTGGTGGTG CTGCTGACCA GAGCCCTGAG 151 AGCCTGCTGC AGCTGAAGGC CCTGAAGCCA GGTGTCATCC AGATTCTTGG 201 TGTGAAGACC AGCCGCTTCC TCTGCCAGAG GCCAGATGGG GCCCTCTATG 251 GCAGCCTTCA TTTTGATCCT GAGGCCTGCA GCTTCCGGGA GCGGCTGCTG 301 GAAGATGGCT ACAATGTTTA TCAAAGTGAA GCACATGGCC TGCCCCTGCA 351 CCTTCCTGGA AATAAAAGCC CCCACAGAGA TCCTGCCCCC CGGGGCCCTG 401 CCCGCTTCCT GCCCCTGCCT GGGCTGCCCC CGGCCCTGCC TGAGCCGCCG 451 GGCATCCTGG CCCCCCAGCC CCCAGATGTG GGCAGCAGTG ACCCCCTCTC 501 CATGGTTGGT GGCAGCCAGG GCCGCAGCCC CAGCTATGAA AGT Coding sequence of FGF21 variant 2 SEQ ID NO: 77 1 CACCCCATCC CAGACAGCAG CCCGCTGCTG CAGTTTGGTG GTCAAGTTCG 51 ACAGCGCTAC CTCTACACAG ATGATGCCCA GCAGACAGAA GCTCATTTAG 101 AAATAAGAGA AGATGGAACT GTTGGTGGGG CGGCGGACCA GTCTCCAGAA 151 AGTTTATTAC AATTAAAAGC TTTAAAACCT GGTGTCATCC AGATTCTTGG 201 TGTCAAGACC AGCCGCTTCC TCTGCCAGAG GCCAGATGGT GCTTTATATG 251 GCAGCCTTCA TTTTGATCCT GAGGCCTGCT CCTTCCGAGA AAGATTATTA 301 GAAGATGGCT ACAATGTCTA CCAGAGTGAA GCTCATGGTC TTCCTCTTCA 351 TCTTCCTGGG AACAAGTCGC CGCACCGGGA CCCGGCGCCG CGGGGCCCGG 401 CGCGCTTCCT GCCGCTGCCG GGCCTCCCGC CGGCGCTGCC GGAGCCGCCG 451 GGCATCCTGG CGCCGCAGCC GCCGGATGTG GGCAGCAGTG ACCCCCTCTC 501 CATGGTTGGT GGCAGCCAGG GCCGCTCGCC CAGCTATGAA AGT Coding sequence of FGF21 variant 2 SEQ ID NO: 78 1 CACCCCATCC CGGACTCGTC GCCGCTGCTG CAGTTTGGTG GTCAAGTTCG 51 ACAGCGCTAC CTCTACACAG ATGATGCTCA ACAAACAGAA GCTCATTTAG 101 AAATAAGAGA AGATGGAACT GTTGGTGGGG CGGCGGACCA GTCGCCGGAG 151 AGTTTATTAC AATTAAAAGC TTTAAAACCT GGTGTCATCC AGATTCTTGG 201 TGTCAAGACC AGCCGCTTCC TCTGCCAGAG GCCAGATGGT GCTTTATATG 251 GAAGTTTACA TTTTGACCCG GAGGCCTGCT CCTTCCGAGA AAGATTATTA 301 GAAGATGGCT ACAATGTTTA TCAAAGTGAA GCTCATGGTC TTCCTCTTCA 351 TCTTCCTGGG AACAAGTCGC CGCACCGGGA CCCGGCGCCG CGGGGCCCGG 401 CGCGCTTCCT CCCGCTGCCG GGCCTCCCGC CGGCGCTGCC GGAGCCGCCG 451 GGCATCCTGG CGCCGCAGCC GCCGGATGTT GGCTCCTCGG ACCCGCTCTC 501 CATGGTTGGT GGCAGCCAGG GCCGCTCGCC CAGCTATGAA AGT Coding sequence of FGF21 variant 2 SEQ ID NO: 79 1 CACCCCATCC CGGACAGCAG CCCTTTATTA CAATTTGGTG GTCAAGTAAG 51 ACAAAGATAT TTATATACAG ATGATGCACA ACAAACAGAA GCACATTTAG 101 AAATAAGAGA AGATGGAACT GTTGGTGGGG CGGCGGACCA AAGTCCTGAA 151 AGTTTATTAC AATTAAAAGC ATTAAAACCT GGTGTTATTC AAATATTAGG 201 AGTAAAAACC AGCCGCTTCC TCTGCCAAAG ACCAGATGGT GCTTTATATG 251 GAAGTTTACA TTTTGATCCT GAGGCCTGCT CCTTCAGAGA AAGATTATTA 301 GAAGATGGTT ATAATGTTTA TCAAAGTGAA GCACATGGTC TTCCTTTACA 351 TTTACCTGGA AATAAATCCC CGCACCGGGA CCCGGCGCCG CGGGGCCCGG 401 CGCGCTTCCT CCCGCTGCCG GGCCTCCCGC CGGCGCTGCC GGAGCCGCCG 451 GGCATCCTGG CGCCGCAGCC GCCGGATGTT GGCAGCAGTG ACCCCCTCTC 501 CATGGTTGGT GGCAGCCAAG GCCGCTCGCC CAGCTATGAA AGT FGF21 is a 2-month-old SEQ ID NO: 80 1 CACCCCATCC CAGACAGCAG CCCCCTGCTG CAGTTTGGTG GCCAGGTGCG 51 GCAGCGCTAC CTCTACACAG ATGATGCCCA GCAGACAGAA GCACATTTAG 101 AAATAAGAGA AGATGGAACT GTTGGTGGGG CGGCGGACCA GAGCCCAGAA 151 AGTTTATTAC AATTAAAAGC ATTAAAACCT GGTGTCATCC AGATTCTTGG 201 AGTAAAAACC AGCCGCTTCC TCTGCCAGAG GCCAGATGGG GCCCTCTATG 251 GCAGCCTTCA TTTTGATCCT GAGGCCTGCA GCTTCCGGGA GCGGCTGCTG 301 GAAGATGGCT ACAATGTTTA TCAAAGTGAA GCACATGGCC TGCCCCTGCA 351 CCTTCCTGGA AATAAAAGCC CCCACCGGGA CCCGGCGCCC CGGGGCCCGG 401 CGCGCTTCCT GCCCCTGCCT GGGCTGCCGC CGGCGCTGCC TGAGCCGCCG 451 GGCATCCTGG CGCCCCAGCC CCCAGATGTG GGCAGCAGTG ACCCCCTCTC 501 CATGGTTGGT GGCAGCCAGG GCCGCAGCCC CAGCTATGAA AGT FGF21 is a 2-month-old SEQ ID NO: 81 1 CACCCCATCC CAGACAGCAG CCCCCTGCTG CAGTTTGGTG GCCAGGTGCG 51 GCAGCGCTAC CTCTACACAG ATGATGCCCA GCAAACAGAA GCACATTTAG 101 AAATAAGAGA AGATGGAACT GTTGGTGGGG CGGCGGACCA GTCTCCAGAA 151 AGTTTATTAC AATTAAAAGC ATTAAAACCT GGTGTCATTC AAATATTAGG 201 AGTAAAAACC AGCCGCTTCC TCTGCCAGAG GCCAGATGGT GCTTTATATG 251 GAAGTTTACA TTTTGATCCT GAGGCCTGCT CCTTCAGAGA AAGATTATTA 301 GAAGATGGCT ACAATGTTTA TCAAAGTGAA GCACATGGTC TTCCTCTTCA 351 TCTTCCTGGA AATAAATCTC CCCACCGGGA CCCGGCGCCG CGGGGCCCGG 401 CGCGCTTCCT GCCCCTTCCT GGGCTGCCGC CGGCGCTGCC GGAGCCGCCG 451 GGCATCCTGG CGCCCCAGCC GCCGGATGTT GGCAGCAGTG ACCCCCTCTC 501 CATGGTTGGT GGCAGCCAGG GCCGCAGCCC CAGCTATGAA AGT Coding sequence of FGF21 variant 2 SEQ ID NO: 82 1 CACCCCATCC CAGACAGCAG CCCCCTGCTG CAGTTTGGTG GCCAGGTGCG 51 GCAGCGCTAC CTCTACACAG ATGATGCCCA GCAGACAGAA GCTCATTTAG 101 AAATAAGAGA AGATGGAACT GTTGGTGGTG CTGCTGACCA GAGCCCTGAG 151 AGCCTGCTGC AGCTGAAGGC CCTGAAGCCA GGTGTCATCC AGATTCTTGG 201 AGTAAAAACC AGCCGCTTCC TCTGCCAGAG GCCAGATGGG GCCCTCTATG 251 GCAGCCTTCA TTTTGATCCT GAGGCCTGCA GCTTCCGGGA GCGGCTGCTG 301 GAAGATGGCT ACAATGTTTA TCAAAGTGAA GCTCATGGCC TGCCCCTGCA 351 CCTTCCTGGA AATAAAAGCC CCCACAGAGA TCCTGCCCCC CGGGGCCCTG 401 CCCGCTTCCT GCCCCTGCCT GGGCTGCCCC CGGCCCTGCC TGAGCCGCCG 451 GGCATCCTGG CCCCCCAGCC CCCAGATGTG GGCAGCAGTG ACCCCCTCTC 501 CATGGTTGGT GGCAGCCAGG GCCGCAGCCC CAGCTATGAA AGT Coding sequence of FGF21 variant 2 SEQ ID NO: 83 1 CACCCTATCC CTGACAGCAG CCCTCTGCTG CAGTTCGGCG GCCAGGTGAG 51 ACAGAGATAC CTGTACACCG ACGACGCCCA GCAGACCGAG GCCCACCTGG 101 AGATCAGAGA GGACGGCACC GTGGGCGGCG CCGCCGACCA GAGCCCTGAG 151 AGCCTGCTGC AGCTGAAGGC CCTGAAGCCT GGCGTGATCC AGATCCTGGG 201 CGTGAAGACC AGCAGATTCC TGTGCCAGAG ACCTGACGGC GCCCTGTACG 251 GCAGCCTGCA CTTCGACCCT GAGGCCTGCA GCTTCAGAGA GAGACTGCTG 301 GAGGACGGCT ACAACGTGTA CCAGAGCGAG GCCCACGGCC TGCCTCTGCA 351 CCTGCCTGGC AACAAGAGCC CTCACAGAGA CCCTGCCCCT AGAGGCCCTG 401 CCAGATTCCT GCCTCTGCCT GGCCTGCCTC CTGCCCTGCC TGAGCCTCCT 451 GGCATCCTGG CCCCTCAGCC TCCTGACGTG GGCAGCAGCG ACCCTCTGAG 501 CATGGTGGGC GGCAGCCAGG GCAGAAGCCC TAGCTACGAG AGC Coding sequence of FGF21 variant 2 SEQ ID NO: 84 1 CACCCCATCC CAGACAGCAG CCCACTGCTC CAGTTTGGAG GGCAGGTTCG 51 GCAGCGCTAC CTGTATACAG ATGATGCCCA GCAGACAGAA GCCCACCTGG 101 AGATCCGGGA GGACGGCACC GTGGGAGGAG CTGCTGACCA GAGCCCTGAA 151 AGCCTCCTGC AGCTGAAAGC ACTGAAGCCT GGGGTCATCC AGATCCTGGG 201 GGTGAAAACT TCCCGCTTCC TGTGCCAGAG GCCTGATGGG GCCCTCTATG 251 GAAGTCTTCA TTTTGATCCT GAGGCCTGCT CCTTCAGAGA AAGACTTCTG 301 GAAGATGGCT ACAATGTGTA CCAGAGTGAA GCTCATGGCC TGCCCCTGCA 351 TCTGCCTGGC AACAAGTCTC CCCACCGGGA CCCAGCTCCC CGGGGCCCTG 401 CTAGATTTCT TCCTCTTCCA GGCCTCCCTC CAGCCCTGCC GGAGCCTCCT 451 GGAATTCTGG CCCCTCAGCC ACCAGATGTG GGCTCCTCTG ACCCTCTGTC 501 CATGGTGGGG GGCAGCCAGG GCCGCAGCCC CTCCTATGAA AGT Coding sequence of FGF21 variant 2 SEQ ID NO: 85 1 CACCCCATCC CAGACAGCAG CCCACTGCTC CAGTTTGGAG GGCAGGTTCG 51 GCAGCGCTAC CTGTATACAG ATGATGCCCA GCAGACAGAA GCCCACCTGG 101 AGATCCGGGA GGACGGCACC GTGGGAGGAG CTGCTGACCA GAGCCCTGAA 151 AGCCTCCTGC AGCTGAAAGC ACTGAAGCCT GGGGTCATCC AGATCCTGGG 201 GGTGAAAACT TCCCGCTTCC TGTGCCAGAG GCCTGATGGG GCCCTCTATG 251 GAAGTCTTCA TTTTGATCCT GAGGCCTGCT CCTTCAGAGA AAGACTTCTG 301 GAAGATGGCT ACAATGTGTA CCAGAGTGAA GCTCATGGCC TGCCCCTGCA 351 TCTGCCTGGC AACAAGTCTC CCCACCGGGA CCCAGCTCCC CGGGGCCCTG 401 CTAGATTTCT TCCTCTTCCA GGCCTCCCTC CAGCCCTGCC GGAGCCTCCT 451 GGAATTCTGG CCCCTCAGCC ACCAGATGTG GGCTCCTCTG ACCCTCTGTC 501 CATGGTGGGG GGCAGCCAGG GCCGCAGCCC CTCCTATGAA AGT Coding sequence of FGF21 variant 2 SEQ ID NO: 86 1 CACCCCATCC CAGACAGCAG CCCACTGCTC CAGTTTGGAG GGCAGGTTCG 51 GCAGCGCTAC CTGTATACAG ATGATGCCCA GCAGACAGAA GCCCACCTGG 101 AGATCCGGGA GGACGGCACC GTGGGAGGAG CTGCTGACCA GAGCCCTGAA 151 AGCCTCCTGC AGCTGAAAGC ACTGAAGCCT GGGGTCATCC AGATCCTGGG 201 GGTGAAAACT TCCCGCTTCC TGTGCCAGAG GCCTGATGGG GCCCTCTATG 251 GAAGTCTTCA TTTTGATCCT GAGGCCTGCT CCTTCAGAGA AAGACTTCTG 301 GAAGATGGCT ACAATGTGTA CCAGAGTGAA GCTCATGGCC TGCCCCTGCA 351 TCTGCCTGGC AACAAGTCTC CCCACCGGGA CCCAGCTCCC CGGGGCCCTG 401 CTAGATTTCT TCCTCTTCCA GGCCTCCCTC CAGCCCTGCC GGAGCCTCCT 451 GGAATTCTGG CCCCTCAGCC ACCAGATGTG GGCTCCTCTG ACCCTCTGTC 501 CATGGTGGGG GGCAGCCAGG GCCGCAGCCC CTCCTATGAA AGT Coding sequence of FGF21 variant 2 SEQ ID NO: 87 1 CACCCCATCC CAGACAGCAG CCCACTGCTC CAGTTTGGAG GGCAGGTTCG 51 GCAGCGCTAC CTGTATACAG ATGATGCCCA GCAGACAGAA GCCCACCTGG 101 AGATCCGGGA GGACGGCACC GTGGGAGGAG CTGCTGACCA GAGCCCTGAA 151 AGCCTCCTGC AGCTGAAAGC ACTGAAGCCT GGGGTCATCC AGATCCTGGG 201 GGTGAAAACT TCCCGCTTCC TGTGCCAGAG GCCTGATGGG GCCCTCTATG 251 GAAGTCTTCA TTTTGATCCT GAGGCCTGCT CCTTCAGAGA AAGACTTCTG 301 GAAGATGGCT ACAATGTGTA CCAGAGTGAA GCTCATGGCC TGCCCCTGCA 351 TCTGCCTGGC AACAAGTCTC CCCACCGGGA CCCAGCTCCC CGGGGCCCTG 401 CTAGATTTCT TCCTCTTCCA GGCCTCCCTC CAGCCCTGCC GGAGCCTCCT 451 GGAATTCTGG CCCCTCAGCC ACCAGATGTG GGCTCCTCTG ACCCTCTGTC 501 CATGGTGGGG GGCAGCCAGG GCCGCAGCCC CTCCTATGAA AGT Coding sequence of FGF21 variant 2 SEQ ID NO: 88 1 CACCCCATCC CAGACAGCAG CCCACTGCTC CAGTTTGGAG GGCAGGTTCG 51 GCAGCGCTAC CTGTATACAG ATGATGCCCA GCAGACAGAA GCCCACCTGG 101 AGATCCGGGA GGACGGCACC GTGGGAGGAG CTGCTGACCA GAGCCCTGAA 151 AGCCTCCTGC AGCTGAAAGC ACTGAAGCCT GGGGTCATCC AGATCCTGGG 201 GGTGAAAACT TCCCGCTTCC TGTGCCAGAG GCCTGATGGG GCCCTCTATG 251 GCTCTCTTCA TTTTGATCCT GAGGCCTGCT CCTTCAGAGA AAGACTTCTG 301 GAAGATGGCT ACAATGTGTA CCAGAGTGAA GCTCATGGCC TGCCCCTGCA 351 TCTGCCTGGC AACAAGTCTC CCCACCGGGA CCCAGCTCCC CGGGGCCCTG 401 CTAGATTTCT TCCTCTTCCA GGCCTCCCTC CAGCCCTGCC GGAGCCTCCT 451 GGAATTCTGG CCCCTCAGCC ACCAGATGTG GGCTCCTCTG ACCCTCTGTC 501 CATGGTGGGG GGCAGCCAGG GCCGCAGCCC CTCCTATGAA AGT Coding sequence of FGF21 variant 2 SEQ ID NO: 89 1 CACCCCATCC CAGACAGCAG CCCCCTGCTG CAGTTTGGTG GCCAGGTGCG 51 GCAGCGCTAC CTCTACACAG ATGATGCCCA GCAGACAGAG GCCCACCTGG 101 AGATCAGAGA AGATGGAACA GTTGGAGGAG CTGCTGACCA GAGCCCTGAG 151 AGCCTGCTGC AGCTGAAGGC CCTGAAGCCT GGGGTCATCC AGATCCTGGG 201 CGTGAAGACC AGCCGCTTCC TCTGCCAGAG GCCAGATGGG GCCCTGTATG 251 GCAGCCTGCA CTTTGATCCT GAGGCCTGCT CCTTCCGGGA GCGGCTGCTG 301 GAGGATGGCT ACAATGTCTA CCAGAGTGAA GCTCATGGCC TGCCCCTGCA 351 CCTGCCTGGG AACAAGAGCC CCCACCGGGA CCCTGCCCCC CGGGGCCCTG 401 CCCGCTTCCT GCCCCTGCCT GGGCTGCCCC CAGCCCTGCC TGAGCCCCCA 451 GGCATCCTGG CCCCCCAGCC CCCAGATGTG GGCAGCAGTG ACCCCCTGTC 501 CATGGTGGGC GGCAGCCAGG GCCGCAGCCC CAGCTATGAG AGC Coding sequence of FGF21 variant 2 SEQ ID NO: 90 1 CACCCCATCC CAGACAGCAG CCCCCTGCTG CAGTTTGGTG GCCAGGTGCG 51 GCAGCGCTAC CTCTACACAG ATGATGCCCA GCAGACAGAA GCACATTTAG 101 AAATAAGAGA AGATGGAACT GTTGGTGGTG CTGCTGACCA GAGCCCTGAG 151 AGCCTGCTGC AGCTGAAGGC CCTGAAGCCA GGTGTCATCC AGATTCTTGG 201 TGTGAAGACC AGCCGCTTCC TCTGCCAGAG GCCAGATGGG GCCCTCTATG 251 GCAGCCTTCA TTTTGATCCT GAGGCCTGCA GCTTCCGGGA GCGGCTGCTG 301 GAAGATGGCT ACAATGTTTA TCAAAGTGAA GCTCATGGCC TGCCCCTGCA 351 CCTTCCTGGA AATAAAAGCC CCCACAGAGA TCCTGCCCCC CGGGGCCCTG 401 CCCGCTTCCT GCCCCTGCCT GGGCTGCCCC CGGCCCTGCC TGAGCCGCCG 451 GGCATCCTGG CCCCCCAGCC CCCAGATGTG GGCAGCAGTG ACCCCCTCTC 501 CATGGTTGGT GGCAGCCAGG GCCGCAGCCC CAGCTATGAA AGT Coding sequence of FGF21 variant 2 SEQ ID NO: 91 1 CACCCCATCC CAGACAGCAG CCCCCTGCTG CAGTTTGGTG GCCAGGTGCG 51 GCAGCGCTAC CTCTACACAG ATGATGCCCA GCAGACAGAG GCCCACCTGG 101 AGATCAGAGA AGATGGAACA GTTGGTGGTG CTGCTGACCA GAGCCCTGAG 151 AGCCTGCTGC AGCTGAAGGC CCTGAAGCCT GGTGTCATCC AGATCCTGGG 201 TGTGAAGACC AGCCGCTTCC TCTGCCAGAG GCCAGATGGG GCCCTCTATG 251 GCAGCCTGCA CTTTGATCCT GAGGCCTGCT CCTTCCGGGA GCGGCTGCTG 301 GAGGATGGCT ACAATGTCTA CCAGAGTGAA GCTCATGGCC TGCCCCTGCA 351 CCTGCCTGGG AACAAGAGCC CCCACCGGGA CCCCGCGCCC CGGGGCCCTG 401 CCCGCTTCCT GCCCCTGCCT GGGCTGCCGC CGGCCCTGCC TGAGCCGCCG 451 GGCATCCTGG CCCCCCAGCC CCCAGATGTG GGCAGCAGTG ACCCCCTCTC 501 CATGGTGGGC GGCAGCCAGG GCCGCAGCCC CAGCTATGAA AGT Coding sequence of FGF21 variant 2 SEQ ID NO: 92 1 CACCCCATCC CAGACAGCAG CCCCCTGCTG CAGTTTGGTG GCCAGGTGCG 51 GCAGCGCTAC CTCTACACAG ATGATGCCCA GCAGACAGAA GCCCACCTGG 101 AAATAAGAGA AGATGGAACT GTTGGTGGTG CTGCTGACCA GAGCCCTGAG 151 AGCCTGCTGC AGCTGAAGGC CCTGAAGCCA GGTGTCATCC AGATTCTTGG 201 TGTGAAGACC AGCCGCTTCC TCTGCCAGAG GCCAGATGGG GCCCTCTATG 251 GCAGCCTGCA CTTTGATCCT GAGGCCTGCA GCTTCCGGGA GCGGCTGCTG 301 GAAGATGGCT ACAATGTCTA CCAGAGTGAA GCTCATGGCC TGCCCCTGCA \351 CCTGCCTGGA AATAAAAGCC CCCACAGAGA TCCTGCCCCC CGGGGCCCTG[[ID=](16) 401 CCCGCTTCCT GCCCCTGCCT GGGCTGCCCC CAGCCCTGCC TGAGCCCCCG 451 GGCATCCTGG CCCCCCAGCC CCCAGATGTG GGCAGCAGTG ACCCCCTCTC 501 CATGGTGGGG GGCAGCCAGG GCCGCAGCCC CAGCTATGAA AGT Coding sequence of FGF21 variant 2 SEQ ID NO: 93 1 CACCCTATCC CTGACAGCAG CCCTCTGCTG CAGTTCGGCG GCCAGGTGAG 51 ACAGAGATAC CTGTACACCG ACGACGCCCA GCAGACCGAG GCCCACCTGG 101 AGATCAGAGA GGACGGCACC GTGGGCGGCG CCGCCGACCA GAGCCCTGAG 151 AGCCTGCTGC AGCTGAAGGC CCTGAAGCCT GGCGTGATCC AGATCCTGGG 201 CGTGAAGACC AGCAGATTCC TGTGCCAGAG ACCTGACGGC GCCCTGTACG 251 GCAGCCTGCA CTTCGACCCT GAGGCCTGCA GCTTCAGAGA GAGACTGCTG 301 GAGGACGGCT ACAACGTGTA CCAGAGCGAG GCCCACGGCC TGCCTCTGCA 351 CCTGCCTGGC AACAAGAGCC CTCACAGAGA CCCTGCCCCT AGAGGCCCTG 401 CCAGATTCCT GCCTCTGCCT GGCCTGCCTC CTGCCCTGCC TGAGCCTCCT 451 GGCATCCTGG CCCCTCAGCC TCCTGACGTG GGCAGCAGCG ACCCTCTGAG 501 CATGGTGGGC GGCAGCCAGG GCAGAAGCCC TAGCTACGAG AGC Coding sequence of FGF21 variant 2 SEQ ID NO: 94 1 CACCCCATCC CAGACAGCAG CCCCCTGCTG CAGTTTGGGG GCCAGGTGCG 51 GCAGCGCTAC CTCTACACAG ATGATGCCCA GCAGACAGAG GCCCACCTGG 101 AGATCAGAGA AGATGGAACA GTTGGAGGAG CTGCTGACCA GAGCCCTGAG 151 AGCCTGCTGC AGCTGAAGGC CCTGAAGCCT GGGGTCATCC AGATCCTGGG 201 CGTGAAGACC AGCCGCTTCC TCTGCCAGAG GCCAGATGGG GCCCTGTATG 251 GCAGCCTGCA CTTTGATCCT GAGGCCTGCA GCTTCCGGGA GCGGCTGCTG 301 GAAGATGGCT ACAATGTCTA CCAGAGTGAG GCCCATGGCC TGCCCCTGCA 351 CCTGCCTGGA AATAAAAGCC CCCACAGAGA TCCTGCCCCC CGGGGCCCTG 401 CCCGCTTCCT GCCCCTGCCT GGCCTGCCCC CAGCCCTGCC TGAGCCCCCA 451 GGCATCCTGG CCCCCCAGCC CCCAGATGTG GGCAGCAGTG ACCCCCTGTC 501 CATGGTGGGC GGCAGCCAGG GCCGCAGCCC CAGCTATGAA AGT Coding sequence of FGF21 variant 2 SEQ ID NO: 95 1 CACCCCATCC CAGACAGCAG CCCCCTGCTG CAGTTTGGTG GCCAGGTGCG 51 GCAGCGCTAC CTCTACACAG ATGATGCCCA GCAGACAGAG GCCCACCTGG 101 AGATCAGAGA AGATGGAACA GTTGGTGGTG CTGCTGACCA GAGCCCTGAG 151 AGCCTGCTGC AGCTGAAGGC CCTGAAGCCT GGGGTCATCC AGATCCTGGG 201 CGTGAAGACC AGCCGCTTCC TCTGCCAGAG GCCAGATGGG GCCCTGTATG 251 GCAGCCTGCA CTTTGATCCT GAGGCCTGCT CCTTCCGGGA GCGGCTGCTG 301 GAGGATGGCT ACAATGTCTA CCAGAGTGAA GCTCATGGCC TGCCCCTGCA 351 CCTGCCTGGG AACAAGAGCC CCCACCGGGA CCCTGCCCCC CGGGGCCCTG 401 CCCGCTTCCT GCCCCTGCCT GGGCTGCCCC CAGCCCTGCC TGAGCCCCCA 451 GGCATCCTGG CCCCCCAGCC CCCAGATGTG GGCAGCAGTG ACCCCCTGTC 501 CATGGTGGGC GGCAGCCAGG GCCGCAGCCC CAGCTATGAA AGT Coding sequence of FGF21 variant 2 SEQ ID NO: 96 1 CACCCCATCC CAGACAGCAG CCCCCTGCTG CAGTTTGGTG GCCAGGTGCG 51 GCAGCGCTAC CTCTACACAG ATGATGCCCA GCAGACAGAA GCACATTTAG 101 AAATAAGAGA AGATGGAACT GTTGGTGGGG CGGCGGACCA GAGCCCTGAG 151 AGCCTGCTGC AGCTGAAGGC CTTGAAACCT GGTGTCATCC AGATTCTTGG 201 AGTAAAAACC AGCCGCTTCC TCTGCCAGAG GCCAGATGGG GCCCTCTATG 251 GCAGCCTTCA TTTTGATCCT GAGGCCTGCA GCTTCCGGGA GCGGCTGCTG 301 GAAGATGGCT ACAATGTTTA TCAAAGTGAA GCACATGGCC TGCCCCTGCA 351 CCTTCCTGGA AATAAAAGCC CCCACCGGGA CCCGGCGCCC CGGGGCCCGG 401 CCCGCTTCCT GCCCCTGCCT GGGCTGCCCC CGGCGCTGCC TGAGCCGCCG 451 GGCATCCTGG CCCCCCAGCC CCCAGATGTG GGCAGCAGTG ACCCCCTCTC 501 CATGGTTGGT GGCAGCCAGG GCCGCAGCCC CAGCTATGAA AGT Coding sequence of FGF21 variant 2 SEQ ID NO: 97 1 CACCCCATCC CAGACAGCAG CCCCCTGCTG CAGTTTGGTG GCCAGGTGCG 51 GCAGCGCTAC CTCTACACAG ATGATGCCCA GCAGACGGAG GCCCACCTGG 101 AGATCCGAGA AGATGGAACT GTTGGTGGCG CGGCGGACCA GAGCCCTGAG 151 AGCCTGCTGC AGCTGAAGGC CCTGAAGCCA GGTGTCATCC AGATTCTTGG 201 TGTCAAGACC AGCCGCTTCC TCTGCCAGAG GCCAGATGGG GCCCTCTATG 251 GCAGCCTTCA TTTTGATCCT GAGGCCTGCT CCTTCCGGGA GCGGCTGCTG 301 GAGGATGGCT ACAATGTCTA CCAGAGTGAA GCTCATGGAC TTCCTCTTCA 351 TCTTCCTGGG AACAAGAGCC CCCACCGGGA CCCGGCGCCG CGGGGCCCGG 401 CGCGCTTCCT GCCCCTGCCT GGGCTGCCGC CGGCGCTGCC TGAGCCGCCG 451 GGCATCCTGG CGCCCCAGCC CCCAGATGTG GGCAGCAGTG ACCCCCTCTC 501 CATGGTTGGT GGCAGCCAGG GCCGCAGCCC CAGCTATGAA AGT Coding sequence of FGF21 variant 2 SEQ ID NO: 98 1 CACCCCATCC CAGACAGCAG CCCCCTGCTG CAGTTTGGTG GCCAGGTGCG 51 GCAGCGCTAC CTCTACACAG ATGATGCCCA GCAGACAGAG GCCCACCTGG 101 AGATCAGAGA AGATGGAACA GTTGGTGGTG CTGCTGACCA GAGCCCTGAG 151 AGCCTGCTGC AGCTGAAGGC CCTGAAGCCA GGTGTCATCC AGATCCTGGG 201 TGTGAAGACC AGCCGCTTCC TCTGCCAGAG GCCAGATGGG GCCCTCTATG 251 GCAGCCTGCA CTTTGATCCT GAGGCCTGCA GCTTCCGGGA GCGGCTGCTG 301 GAGGATGGCT ACAATGTCTA CCAGAGTGAA GCTCATGGCC TGCCCCTGCA 351 CCTGCCTGGG AACAAGAGCC CCCACCGGGA CCCGGCGCCC CGGGGCCCGG 401 CGCGCTTCCT GCCCCTGCCT GGGCTGCCGC CGGCGCTGCC TGAGCCGCCG 451 GGCATCCTGG CCCCCCAGCC CCCAGATGTG GGCAGCAGTG ACCCCCTCTC 501 CATGGTGGGC GGCAGCCAGG GCCGCAGCCC CAGCTATGAA AGT Coding sequence of FGF21 variant 2 SEQ ID NO: 99 1 CACCCTATCC CTGACAGCAG CCCTCTGCTG CAGTTCGGCG GCCAGGTGAG 51 ACAGAGATAC CTGTACACCG ACGACGCCCA GCAGACCGAG GCCCACCTGG 101 AGATCAGAGA GGACGGCACC GTGGGCGGCG CCGCCGACCA GAGCCCTGAG 151 AGCCTGCTGC AGCTGAAGGC CCTGAAGCCT GGCGTGATCC AGATCCTGGG 201 CGTGAAGACC AGCAGATTCC TGTGCCAGAG ACCTGACGGC GCCCTGTACG 251 GCAGCCTGCA CTTCGACCCT GAGGCCTGCA GCTTCAGAGA GAGACTGCTG 301 GAGGACGGCT ACAACGTGTA CCAGAGCGAG GCCCACGGCC TGCCTCTGCA 351 CCTGCCTGGC AACAAGAGCC CTCACAGAGA CCCTGCCCCT AGAGGCCCTG 401 CCAGATTCCT GCCTCTGCCT GGCCTGCCTC CTGCCCTGCC TGAGCCTCCT 451 GGCATCCTGG CCCCTCAGCC TCCTGACGTG GGCAGCAGCG ACCCTCTGAG 501 CATGGTGGGC GGCAGCCAGG GCAGAAGCCC TAGCTACGAG AGC Coding sequence of FGF21 variant 2 SEQ ID NO: 100 1 CACCCCATCC CAGACAGCAG CCCCCTGCTG CAGTTTGGTG GCCAGGTGCG 51 GCAGCGCTAC CTCTACACAG ATGATGCCCA GCAGACAGAA GCCCACCTGG 101 AAATAAGAGA AGATGGAACT GTTGGTGGTG CTGCTGACCA GAGCCCTGAG 151 AGCCTGCTGC AGCTGAAGGC CCTGAAGCCA GGTGTCATCC AGATTCTTGG 201 TGTGAAGACC AGCCGCTTCC TCTGCCAGAG GCCAGATGGG GCCCTCTATG 251 GCAGCCTTCA TTTTGATCCT GAGGCCTGCA GCTTCCGGGA GCGGCTGCTG 301 GAAGATGGCT ACAATGTTTA TCAAAGTGAA GCACATGGCC TGCCCCTGCA 351 CCTGCCTGGA AATAAAAGCC CCCACAGAGA TCCTGCCCCC CGGGGCCCTG 401 CCCGCTTCCT GCCCCTGCCT GGGCTGCCCC CGGCCCTGCC TGAGCCGCCG 451 GGCATCCTGG CCCCCCAGCC CCCAGATGTG GGCAGCAGTG ACCCCCTCTC 501 CATGGTTGGT GGCAGCCAGG GCCGCAGCCC CAGCTATGAA AGT Coding sequence of FGF21 variant 2 SEQ ID NO: 101 1 CACCCTATCC CTGACAGCAG CCCTCTGCTG CAGTTCGGCG GCCAGGTGAG 51 ACAGAGATAC CTGTACACCG ACGACGCCCA GCAGACCGAG GCCCACCTGG 101 AGATCAGAGA GGACGGCACC GTGGGCGGCG CCGCCGACCA GAGCCCTGAG 151 AGCCTGCTGC AGCTGAAGGC CCTGAAGCCT GGCGTGATCC AGATCCTGGG 201 CGTGAAGACC AGCAGATTCC TGTGCCAGAG ACCTGACGGC GCCCTGTACG 251 GCAGCCTGCA CTTCGACCCT GAGGCCTGCA GCTTCAGAGA GAGACTGCTG 301 GAGGACGGCT ACAACGTGTA CCAGAGCGAG GCCCACGGCC TGCCTCTGCA 351 CCTGCCTGGC AACAAGAGCC CTCACAGAGA CCCTGCCCCT AGAGGCCCTG 401 CCAGATTCCT GCCTCTGCCT GGCCTGCCTC CTGCCCTGCC TGAGCCTCCT 451 GGCATCCTGG CCCCTCAGCC TCCTGACGTG GGCAGCAGCG ACCCTCTGAG 501 CATGGTGGGC GGCAGCCAGG GCAGAAGCCC TAGCTACGAG AGC Coding sequence of FGF21 variant 2 SEQ ID NO: 102 1 CACCCCATCC CAGACAGCAG CCCCCTGCTG CAGTTTGGTG GCCAGGTGCG 51 GCAGCGCTAC CTCTACACAG ATGATGCCCA GCAGACAGAG GCCCACCTGG 101 AGATCAGAGA AGATGGAACT GTTGGTGGCG CGGCGGACCA GAGCCCTGAG 151 AGCCTGCTGC AGCTGAAGGC CCTGAAGCCA GGTGTCATCC AGATCCTGGG 201 GGTCAAGACC AGCCGCTTCC TCTGCCAGAG GCCAGATGGG GCCCTCTATG 251 GCAGCCTGCA CTTTGATCCT GAGGCCTGCT CCTTCCGGGA GCGGCTGCTG 301 GAGGATGGCT ACAATGTCTA CCAGAGTGAA GCTCATGGCC TGCCCCTGCA 351 CCTTCCTGGG AACAAGAGCC CCCACCGGGA CCCGGCGCCG CGGGGCCCGG 401 CGCGCTTCCT GCCCCTGCCT GGGCTGCCGC CGGCGCTGCC TGAGCCGCCG 451 GGCATCCTGG CGCCCCAGCC CCCAGATGTG GGCAGCAGTG ACCCCCTCTC 501 CATGGTTGGT GGCAGCCAGG GCCGCAGCCC CAGCTATGAA AGT Coding sequence of FGF21 variant 2 SEQ ID NO: 103 1 CACCCCATCC CAGACAGCAG CCCCCTGCTG CAGTTTGGTG GCCAGGTGCG 51 GCAGCGCTAC CTCTACACAG ATGATGCCCA GCAGACAGAA GCACATTTAG 101 AAATAAGAGA AGATGGAACT GTTGGTGGTG CTGCTGACCA GAGCCCTGAG 151 AGCCTGCTGC AGCTGAAGGC CTTGAAACCT GGTGTCATCC AGATTCTTGG 201 AGTAAAAACC AGCCGCTTCC TCTGCCAGAG GCCAGATGGG GCCCTCTATG 251 GCAGCCTTCA TTTTGATCCT GAGGCCTGCA GCTTCCGGGA GCGGCTGCTG 301 GAAGATGGCT ACAATGTTTA TCAAAGTGAA GCACATGGCC TGCCCCTGCA 351 CCTTCCTGGA AATAAAAGCC CCCACAGAGA TCCTGCCCCC CGGGGCCCTG 401 CCCGCTTCCT GCCCCTGCCT GGGCTGCCCC CGGCCCTGCC TGAGCCGCCG 451 GGCATCCTGG CCCCCCAGCC CCCAGATGTG GGCAGCAGTG ACCCCCTCTC 501 CATGGTTGGT GGCAGCCAGG GCCGCAGCCC CAGCTATGAA AGT Coding sequence of FGF21 variant 2 SEQ ID NO: 104 1 CACCCCATCC CAGACAGCAG CCCCCTGCTG CAGTTTGGGG GCCAGGTGCG 51 GCAGCGCTAC CTCTACACAG ATGATGCCCA GCAGACAGAG GCCCACCTGG 101 AGATCAGAGA AGATGGAACA GTTGGTGGTG CTGCTGACCA GAGCCCTGAG 151 AGCCTGCTGC AGCTGAAGGC CCTGAAGCCT GGGGTCATCC AGATCCTGGG 201 CGTGAAGACC AGCCGCTTCC TCTGCCAGAG GCCAGATGGG GCCCTGTATG 251 GCAGCCTGCA CTTTGATCCT GAGGCCTGCT CCTTCCGGGA GCGGCTGCTG 301 GAGGATGGCT ACAATGTCTA CCAGAGTGAA GCTCATGGCC TGCCCCTGCA 351 CCTGCCTGGG AACAAGAGCC CCCACCGGGA CCCTGCCCCC CGGGGCCCTG 401 CCCGCTTCCT GCCCCTGCCT GGGCTGCCCC CAGCCCTGCC TGAGCCCCCA 451 GGCATCCTGG CCCCCCAGCC CCCAGATGTG GGCAGCAGTG ACCCCCTGTC 501 CATGGTGGGC GGCAGCCAGG GCCGCAGCCC CAGCTATGAA AGT Coding sequence of FGF21 variant 2 SEQ ID NO: 105 1 CACCCCATCC CAGACAGCAG CCCCCTGCTG CAGTTTGGTG GCCAGGTGCG 51 GCAGCGCTAC CTCTACACAG ATGATGCCCA GCAGACAGAG GCCCACCTGG 101 AGATCAGAGA AGATGGAACT GTTGGTGGTG CTGCTGACCA GAGCCCTGAG 151 AGCCTGCTGC AGCTGAAGGC CCTGAAGCCA GGTGTCATCC AGATCCTGGG 201 CGTGAAGACC AGCCGCTTCC TCTGCCAGAG GCCAGATGGG GCCCTCTATG 251 GCAGCCTGCA CTTTGATCCT GAGGCCTGCT CCTTCCGGGA GCGGCTGCTG 301 GAGGATGGCT ACAATGTCTA CCAGAGTGAA GCTCATGGCC TGCCCCTGCA 351 CCTGCCTGGG AACAAGAGCC CCCACCGGGA CCCGGCGCCC CGGGGCCCTG 401 CCCGCTTCCT GCCCCTGCCT GGGCTGCCGC CGGCGCTGCC TGAGCCGCCG 451 GGCATCCTGG CCCCCCAGCC CCCAGATGTG GGCAGCAGTG ACCCCCTCTC 501 CATGGTGGGC GGCAGCCAGG GCCGCAGCCC CAGCTATGAA AGT Coding sequence of FGF2 variant 2 SEQ ID NO: 106 1 CACCCCATCC CAGACAGCAG CCCCCTGCTG CAGTTTGGGG GCCAGGTGCG 51 GCAGCGCTAC CTCTACACAG ATGATGCCCA GCAGACAGAA GCCCACCTGG 101 AGATCAGAGA AGATGGAACT GTTGGAGGAG CTGCTGACCA GAGCCCTGAG 151 AGCCTGCTGC AGCTGAAGGC CCTGAAGCCT GGGGTCATCC AGATCCTGGG 201 GGTGAAGACC AGCCGCTTCC TCTGCCAGAG GCCAGATGGG GCCCTGTATG 251 GCAGCCTGCA CTTTGATCCT GAGGCCTGCA GCTTCCGGGA GCGGCTGCTG 301 GAAGATGGCT ACAATGTCTA CCAGAGTGAA GCTCATGGCC TGCCCCTGCA 351 CCTGCCTGGA AATAAAAGCC CCCACAGAGA TCCTGCCCCC CGGGGCCCTG 401 CCCGCTTCCT GCCCCTGCCT GGGCTGCCCC CAGCCCTGCC TGAGCCCCCA 451 GGCATCCTGG CCCCCCAGCC CCCAGATGTG GGCAGCAGTG ACCCCCTGTC 501 CATGGTGGGC GGCAGCCAGG GCCGCAGCCC CAGCTATGAA AGT Coding sequence of FGF21 variant 2 SEQ ID NO: 107 1 CACCCCATCC CAGACAGCTC CCCACTGCTT CAGTTTGGAG GCCAGGTGCG 51 GCAGCGCTAC CTGTACACAG ATGATGCCCA GCAGACAGAG GCCCACCTGG 101 AGATCCGGGA GGATGGAACA GTGGGAGGAG CTGCTGACCA GAGCCCAGAA 151 AGCCTTCTGC AGCTCAAGGC CCTGAAGCCA GGGGTCATCC AGATCCTCGG 201 TGTGAAGACC AGCAGATTTC TGTGCCAGAG GCCAGATGGG GCCCTCTATG 251 GATCCCTGCA TTTTGATCCT GAAGCCTGCA GCTTCAGAGA AAGACTCCTG 301 GAAGATGGCT ACAACGTGTA CCAGAGTGAA GCTCATGGCC TTCCTCTGCA 351 CCTCCCCGGC AACAAGAGCC CCCACAGGGA CCCTGCCCCT CGGGGCCCAG 401 CCCGCTTCCT GCCCCTCCCT GGGCTGCCTC CTGCTCTGCC AGAGCCCCCA 451 GGCATCCTGG CCCCCCAGCC CCCTGATGTG GGGAGCAGTG ACCCCCTGTC 501 CATGGTGGGC GGCTCCCAGG GCCGCAGCCC TTCCTATGAA AGT Coding sequence of FGF21 variant 2 SEQ ID NO: 108 1 CACCCTATCC CTGACAGCAG CCCTCTGCTG CAGTTCGGCG GCCAGGTGAG 51 ACAGAGATAC CTGTACACCG ACGACGCCCA GCAGACCGAG GCCCACCTGG 101 AGATCAGAGA GGACGGCACC GTGGGCGGCG CCGCCGACCA GAGCCCTGAG 151 AGCCTGCTGC AGCTGAAGGC CCTGAAGCCT GGCGTGATCC AGATCCTGGG 201 CGTGAAGACC AGCAGATTCC TGTGCCAGAG ACCTGACGGC GCCCTGTACG 251 GCAGCCTGCA CTTCGACCCT GAGGCCTGCA GCTTCAGAGA GAGACTGCTG 301 GAGGACGGCT ACAACGTGTA CCAGAGCGAG GCCCACGGCC TGCCTCTGCA 351 CCTGCCTGGC AACAAGAGCC CTCACAGAGA CCCTGCCCCT AGAGGCCCTG 401 CCAGATTCCT GCCTCTGCCT GGCCTGCCTC CTGCCCTGCC TGAGCCTCCT 451 GGCATCCTGG CCCCTCAGCC TCCTGACGTG GGCAGCAGCG ACCCTCTGAG 501 CATGGTGGGC GGCAGCCAGG GCAGAAGCCC TAGCTACGAG AGC Coding sequence of FGF21 variant 2 SEQ ID NO: 109 1 CACCCCATCC CAGACAGCTC CCCACTGCTT CAGTTTGGAG GCCAGGTGCG 51 CCAGCGCTAC CTCTACACAG ATGATGCCCA GCAGACAGAG GCCCACCTGG 101 AGATCCGGGA GGATGGAACA GTGGGGGGAG CTGCTGACCA GAGCCCAGAA 151 AGCCTCCTCC AGCTCAAGGC CCTGAAGCCA GGGGTCATCC AGATCCTCGG 201 TGTGAAGACC AGCAGATTTC TGTGCCAGAG GCCAGATGGG GCCCTCTATG 251 GATCCCTGCA TTTTGATCCT GAAGCCTGCA GCTTCAGAGA AAGACTTCTG 301 GAAGATGGCT ACAACGTGTA CCAGAGTGAA GCTCATGGCC TTCCTCTGCA 351 CCTCCCCGGC AACAAGAGCC CCCACAGGGA CCCTGCCCCT CGGGGCCCAG 401 CCCGCTTCCT GCCCCTCCCT GGGCTGCCTC CTGCTCTGCC AGAGCCCCCA 451 GGCATCCTGG CCCCCCAGCC CCCTGATGTG GGCAGCAGTG ACCCCCTGTC 501 CATGGTGGGA GGAAGCCAGG GCCGCAGCCC TTCCTATGAA AGT Coding sequence of FGF21 variant 2 SEQ ID NO: 110 1 CACCCCATCC CAGACAGCTC CCCACTGCTT CAGTTTGGAG GCCAGGTGCG 51 GCAGCGCTAC CTCTACACAG ATGATGCCCA GCAGACAGAG GCCCACCTGG 101 AGATCCGGGA GGATGGAACA GTGGGTGGAG CTGCTGACCA GAGCCCAGAA 151 AGCCTTCTCC AGCTGAAGGC CCTCAAGCCA GGGGTCATCC AGATCCTCGG 201 TGTGAAGACC AGCAGATTTC TGTGCCAGAG GCCAGATGGG GCCCTGTATG 251 GATCCCTGCA CTTTGATCCT GAAGCCTGCA GCTTCAGAGA AAGACTCCTG 301 GAAGATGGCT ACAACGTGTA CCAGAGTGAA GCTCATGGCC TCCCCCTCCA 351 CCTCCCTGGC AACAAGAGCC CCCACAGGGA CCCTGCCCCT CGGGGCCCAG 401 CCAGGTTCCT GCCTCTGCCC GGCCTTCCTC CTGCTCTGCC AGAGCCCCCA 451 GGCATCCTGG CCCCCCAGCC CCCTGATGTG GGCAGCAGTG ACCCCCTGTC 501 CATGGTGGGG GGCTCCCAGG GCCGGAGCCC TTCCTATGAA AGT Coding sequence of FGF21 variant 2 SEQ ID NO: 111 1 CACCCCATCC CAGACAGCAG CCCACTGCTT CAGTTTGGAG GCCAGGTGCG 51 GCAGAGATAC CTCTACACAG ATGATGCCCA GCAGACAGAG GCCCACCTGG 101 AAATTCGGGA GGATGGAACA GTGGGGGGAG CTGCTGACCA GAGCCCTGAG 151 AGCCTCCTCC AGCTGAAGGC CCTGAAACCT GGTGTCATCC AGATCCTCGG 201 TGTGAAGACC AGCAGATTTC TGTGCCAGAG GCCTGATGGG GCCCTCTATG 251 GATCCCTGCA TTTTGATCCT GAAGCCTGCA GCTTCAGAGA AAGACTGTTG 301 GAAGATGGCT ACAACGTGTA CCAGAGTGAA GCTCATGGCC TCCCTCTGCA 351 CCTCCCAGGC AACAAGAGCC CCCACAGAGA CCCTGCCCCT CGGGGCCCAG 401 CCAGGTTCCT GCCCCTGCCT GGGCTTCCTC CAGCTCTGCC AGAACCTCCT 451 GGCATCCTGG CCCCCCAGCC CCCAGATGTT GGAAGCAGTG ACCCCCTCTC 501 CATGGTGGGC GGCAGCCAGG GAAGATCTCC ATCCTATGAA AGT Coding sequence of FGF21 variant 2 SEQ ID NO: 112 1 CACCCCATCC CAGACAGCAG CCCCCTGCTG CAGTTTGGTG GCCAGGTGCG 51 GCAGCGCTAC CTCTACACAG ATGATGCCCA GCAGACAGAA GCTCATTTAG 101 AAATAAGAGA AGATGGAACT GTTGGTGGGG CGGCGGACCA GAGCCCTGAG 151 AGCCTGCTGC AGCTGAAGGC CCTGAAGCCA GGTGTCATCC AGATCCTGGG 201 GGTCAAGACC AGCCGCTTCC TCTGCCAGAG GCCAGATGGG GCCCTCTATG 251 GCAGCCTTCA TTTTGATCCT GAGGCCTGCT CCTTCCGGGA GCGGCTGCTG 301 GAGGATGGCT ACAATGTCTA CCAGAGTGAA GCTCATGGAC TTCCTCTTCA 351 TCTTCCTGGG AACAAGAGCC CCCACCGGGA CCCGGCGCCG CGGGGCCCGG 401 CGCGCTTCCT GCCCCTGCCT GGGCTGCCGC CGGCGCTGCC TGAGCCGCCG 451 GGCATCCTGG CGCCCCAGCC CCCAGATGTG GGCAGCAGTG ACCCCCTCTC 501 CATGGTTGGT GGCAGCCAGG GCCGCAGCCC CAGCTATGAA AGT Coding sequence of FGF21 variant 3 SEQ ID NO: 113 1 CACCCTATCC CCGACAGCAG CCCCCTGCTG CAGTTCGGCG GCCAAGTGAG 51 ACAGAGATAC CTGTACACCG ACGACGCTCA GCAGACAGAG GCCCATCTGG 101 AAATCAGAGA GGACGGGACC GTGGGCGGGG CTGCCGATCA GAGCCCCGAG 151 AGCCTGCTGC AGCTGAAGGC CCTGAAGCCC GGCGTGATTC AGATCCTGGG 201 CGTGAAGACA AGCAGATTCC TGTGTCAGAG ACCCGACGGC GCCCTGTACG 251 GCAGCCTGCA CTTCGACCCC GAGGCCTGCA GCTTCAGAGA GAGACTGCTG 301 GAGGACGGCT ACAACGTGTA TCAGAGCGAG GCCCACGGCC TGCCCCTCCA 351 CCTGCCCGGG AACAAGAGCC CTCACAGAGA TCCTGCTCCT AGAGGCCCCG 401 CTAGATTTCT CCCCCTGCCC GGGCTGCCTC CCGCCCCTCC TGAACCCCCT 451 GGCATCCTCG CTCCCCAACC CCCTGATGTG GGCAGCAGCG ACCCCCTGAG 501 CATGGTGGGG GGCAGCCAAG GCAGAAGCCC TAGCTACGCT AGC Amino acid sequence of GLP-1 variant 1 SEQ ID NO: 114 HGEGTFTSDVSSYLEEQAAKEFIAWLVKGGG Coding sequence of GLP-1 variant 1 SEQ ID NO: 115 1 CACGGCGAGG GCACCTTCAC CAGCGACGTG AGCAGCTACC TGGAGGAGCA 51 GGCCGCCAAG GAGTTCATCG CCTGGCTGGT GAAGGGCGGC GGC Coding sequence of GLP-1 variant 1 SEQ ID NO: 116 1 CATGGAGAAG GCACCTTCAC CAGTGATGTC AGCAGCTACC TGGAGGAGCA 51 GGCGGCCAAG GAGTTCATTG CCTGGCTGGT GAAAGGTGGT GGT The coding sequence of GLP-1 variant 1 SEQ ID NO: 117 1 CATGGAGAAGGCACCTTCAC CAGTGATGTC AGCAGCTACC TGGAGGAGCA 51 GGCGGCCAAA GAATTTATTG CCTGGCTGGT GAAAGGAGGT GGT The coding sequence of GLP-1 variant 1 SEQ ID NO: 118 1 CATGGAGAGG GCACCTTTCAC CAGTGATGTC AGCAGCTACC TGGAGGAGCA 51 GGCTGCCAAG GAGTTCATTG CCTGGCTGGT GAAAGGAGGA GGA The coding sequence of GLP-1 variant 1 SEQ ID NO: 119 1 CATGGAGAGG GCACCTTTCAC CAGTGATGTC AGCAGCTACC TGGAGGAGCA 51 GGCTGCCAAG GAGTTCATTG CCTGGCTGGT GAAAGGTGGT GGT The coding sequence of GLP-1 variant 1 SEQ ID NO: 120 1 CATGGAGAAGGCACCTTCAC CAGTGATGTC AGCAGCTACC TGGAGGAGCA 51 GGCTGCCAAG GAATTTATTG CCTGGCTGGT GAAAGGAGGA GGA The coding sequence of GLP-1 variant 1 SEQ ID NO: 121 1 CATGGAGAGG GCACCTTTCAC TTCAGATGTC TCCTCCTACT TGGAGGAGCA 51 GGCTGCCAAG GAGTTCATCG CCTGGCTGGT CAAGGGCGGC GGG The coding sequence of GLP-1 variant 1 SEQ ID NO: 122 1 CACGGCGAGG GCACCTTTCAC CAGCGACGTG AGCAGCTACC TGGAGGAGCA 51 GGCCGCCAAG GAGTTCATCG CCTGGCTGGT GAAGGGCGGC GGC The coding sequence of GLP-1 variant 1 SEQ ID NO: 123 1 CATGGAGAGG GCACCTTTCAC TTCAGATGTC AGCAGCTACT TGGAGGAGCA 51 GGCTGCCAAG GAGTTCATCG CCTGGCTGGT CAAGGGCGGG GGT The coding sequence of GLP-1 variant 1 SEQ ID NO: 124 1 CATGGAGAGG GCACCTTTCAC TTCAGATGTC TCCTCCTACT TGGAGGAGCA 51 GGCTGCCAAG GAGTTCATCG CCTGGCTGGT GAAAGGAGGT GGG The coding sequence of GLP-1 variant 1 SEQ ID NO: 125 1 CATGGAGAGG GCACCTTTCAC TTCAGATGTG TCCTCCTACT TGGAGGAGCA 51 GGCTGCCAAG GAGTTCATCG CCTGGCTGGT GAAAGGAGGG GGC The coding sequence of GLP-1 variant 1 SEQ ID NO: 126 1 CATGGAGAAGGCACCTTCAC CAGTGATGTC AGCAGCTACC TGGAGGAGCA 51 GGCGGCCAAG GAGTTCATTG CCTGGCTGGT GAAAGGTGGT GGT The coding sequence of GLP-1 variant 1 SEQ ID NO: 127 1 CATGGAGAAG GCACCTTCAC CAGTGATGTC AGCAGCTATT TAGAAGAACA 51 AGCAGCAAAA GAATTTATTG CCTGGCTGGT GAAAGGTGGT GGT

Claims

1. A circular RNA comprising a regulatory element and an expression element, wherein the expression element comprises a polynucleotide encoding FGF21 or a variant thereof.

2. The circular RNA of claim 1, wherein the regulatory element comprises an internal ribosome entry site (IRES) or a fragment thereof.

3. The circular RNA according to claim 2, wherein the IRES is selected from: Coxsackievirus B3 (CVB3) IRES, Enterovirus 71 (EV71) IRES, Encephalomyocarditis Virus (EMCV) IRES, Picornavir Virus (PV) IRES, Hepatitis C Virus (HCV) IRES, Adenovirus (AdV) IRES, Human Papillomavirus 31 (HPV31) IRES, Human Herpesvirus (HHV) IRES, Raul's Sarcoma Virus (RSV) IRES, Classical Swine Fever Virus (CSFV) IRES, FGF9 IRES, SLC7A1 IRES, and RUNX1 IRES, preferably the IRES is CVB3 IRES (SEQ ID NO: 14).

4. The circular RNA according to any one of the preceding claims, wherein the polynucleotide encoding FGF21 or a variant thereof has at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 98%, at least 99%, or 100% identity with the sequence shown in SEQ ID NO: 15, 16, 17, 18, 19, 20, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, or 113, preferably encoding FGF21 or a variant thereof, as shown in SEQ ID NO: As shown in SEQ ID NO: 15, 16, 17, 18, 19, 20, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112 or 113, more preferably, polynucleotides encoding FGF21 or variants thereof as shown in SEQ ID NO: 17, 76, 89, 95, 101, 107 or 113.

5. The circular RNA according to any one of the preceding claims, wherein the amino acid sequence of FGF21 or a variant thereof is as shown in SEQ ID NO: 1, 2, 3, 4 or 74.

6. The circular RNA according to any one of the preceding claims, wherein the expression element comprises a polynucleotide encoding Fc, ABD, or antialbumin VHH, and optionally the circular RNA comprises a polynucleotide encoding GLP-1 or a variant thereof.

7. The circular RNA of claim 6, wherein the expression element comprises a polynucleotide encoding a linker between Fc and FGF21 or a variant thereof, a polynucleotide encoding a linker between ABD and FGF21 or a variant thereof, or a polynucleotide encoding a linker between antialbumin VHH and FGF21 or a variant thereof.

8. The circular RNA according to claim 7, wherein the amino acid sequence of the adapter is as shown in SEQ ID NO: 70, 71, 72 or 73.

9. The circular RNA according to claim 7 or 8, comprising, from 5' to 3', a polynucleotide encoding Fc (or ABD or antialbumin VHH), a polynucleotide encoding an adapter, and a polynucleotide encoding FGF21 or a variant thereof; Optionally, the circular RNA comprises, from 5' to 3', the following polynucleotides in sequence: a polynucleotide encoding GLP-1 or a variant thereof, a polynucleotide encoding a linker, a polynucleotide encoding Fc (or ABD or antialbumin VHH), a polynucleotide encoding a linker, and a polynucleotide encoding FGF21 or a variant thereof.

10. The circular RNA of claim 6, wherein Fc contains one or more mutations, preferably Fc contains a YTE mutation.

11. The circular RNA according to claim 6, wherein Fc is selected from: IgG1 Fc, IgG2 Fc, IgG3 Fc and IgG4 Fc, preferably Fc is IgG1 Fc.

12. The circular RNA according to claim 6, wherein the amino acid sequence of Fc is as shown in SEQ ID NO: 65, 66 or 67, the amino acid sequence of ABD is as shown in SEQ ID NO: 68, or the amino acid sequence of antialbumin VHH is as shown in SEQ ID NO:

69.

13. The circular RNA of claim 9, wherein the polynucleotide encoding GLP-1 or a variant thereof is as shown in SEQ ID NO: 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126 or 127; and / or the amino acid sequence of GLP-1 or a variant thereof is as shown in SEQ ID NO:

114.

14. The circular RNA according to any one of the preceding claims, comprising a polynucleotide encoding a signal peptide. Optionally, the circular RNA comprises, from 5' to 3', the following polynucleotides in sequence: a polynucleotide encoding a signal peptide, a polynucleotide encoding Fc (or ABD or antialbumin VHH), a polynucleotide encoding an adapter, and a polynucleotide encoding FGF21 or a variant thereof; Optionally, the circular RNA comprises, from 5' to 3', the following polynucleotides in sequence: a polynucleotide encoding a signal peptide, a polynucleotide encoding GLP-1 or a variant thereof, a polynucleotide encoding a linker, a polynucleotide encoding Fc (or ABD or antialbumin VHH), a polynucleotide encoding a linker, and a polynucleotide encoding FGF21 or a variant thereof.

15. The circular RNA according to claim 14, wherein the amino acid sequence of the signal peptide is as shown in SEQ ID NO: 62, 63 or 64.

16. The circular RNA according to any one of the preceding claims, wherein the expression element comprises a sequence having at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 98%, at least 99%, or 100% identity with any sequence shown in SEQ ID NOs: 21-61, preferably the expression element comprises any sequence shown in SEQ ID NOs: 21-61, more preferably the expression element comprises a sequence shown in SEQ ID NOs: 22, 36, 42, 48, 54, 60, or 61, and even more preferably the expression element comprises a sequence shown in SEQ ID NO:

22.

17. A composition comprising the circular RNA of any one of the preceding claims, wherein the composition further comprises a pharmaceutically acceptable excipient.

18. The composition of claim 17, wherein the composition comprises nanoparticles, such as lipid nanoparticles.

19. A method of treating a subject's disease, comprising administering to the subject a therapeutically effective amount of the circular RNA of any one of claims 1-16, or administering to the subject the composition of claim 17 or claim 18.

20. Use of the circular RNA of any one of claims 1-16 or the composition of claim 17 or claim 18 in the preparation of a medicament for treating a disease of a subject.

21. The circular RNA of any one of claims 1-16 or the composition of claim 17 or claim 18, for treating a disease in a subject.

22. The method or use according to any one of claims 19-21, wherein the disease is selected from: diabetes, type 1 diabetes, type 2 diabetes, malnutrition-associated diabetes, diabetic hyperosmolar hyperglycemic state, hypoglycemia in the context of diabetes, diabetic ketoacidosis, diabetic coma, uncontrolled and unstable diabetes.

23. The method or use according to any one of claims 19-21, wherein the disease is selected from: hyperlipoproteinemia, hypercholesterolemia, hypertriglyceridemia, mixed hyperlipidemia, hyperalpha lipoproteinemia, hypolipoproteinemia, hypoalpha lipoproteinemia, and hypobeta lipoproteinemia.

24. The method or use according to any one of claims 19-21, wherein the disease is selected from: obesity or specific nutritional excess and malnutrition.

25. The method or use according to any one of claims 19-21, wherein the disease is selected from: non-alcoholic fatty liver disease, non-alcoholic steatohepatitis (NASH), or hepatic steatosis.

26. The method or use according to any one of claims 19-21, wherein treating the disease is reducing the risk of cardiovascular events.

27. The method or use according to claim 26, wherein the subject is an adult with type 2 diabetes and a confirmed cardiovascular disease.

28. The method or use according to any one of claims 19-21, wherein the disease is selected from: metabolic dysfunction-associated fatty liver disease (MAFLD) or steatohepatitis (MASH).

29. A method for reducing the weight of a subject, comprising administering to the subject a therapeutically effective amount of the circular RNA of any one of claims 1-16, or administering to the subject the composition of claim 17 or claim 18, wherein the method does not reduce the subject's lean muscle mass.

30. A method for reducing fat in a subject, comprising administering to the subject a therapeutically effective amount of the circular RNA of any one of claims 1-16, or administering to the subject the composition of claim 17 or 18, wherein the method does not reduce the subject's lean muscle mass.

31. A method for reducing the weight of a subject, wherein the method comprises administering to the subject a therapeutically effective amount of the circular RNA of any one of claims 1-16, or administering to the subject the composition of claim 17 or claim 18.

32. A method for reducing total cholesterol, LDL-C, or blood glucose in a subject, wherein the method comprises administering to the subject a therapeutically effective amount of the circular RNA of any one of claims 1-16, or administering to the subject the composition of claim 17 or claim 18.

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