WO2020218827A1 - 면역글로불린의 Fc 영역 및 GDF15를 포함하는 융합 폴리펩타이드 - Google Patents
면역글로불린의 Fc 영역 및 GDF15를 포함하는 융합 폴리펩타이드 Download PDFInfo
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- A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
- A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
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- A61K47/6801—Drug-antibody or immunoglobulin conjugates defined by the pharmacologically or therapeutically active agent
- A61K47/6803—Drugs conjugated to an antibody or immunoglobulin, e.g. cisplatin-antibody conjugates
- A61K47/6811—Drugs conjugated to an antibody or immunoglobulin, e.g. cisplatin-antibody conjugates the drug being a protein or peptide, e.g. transferrin or bleomycin
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- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
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- C12N15/09—Recombinant DNA-technology
- C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
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- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/50—Immunoglobulins specific features characterized by immunoglobulin fragments
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- C07K2317/00—Immunoglobulins specific features
- C07K2317/90—Immunoglobulins specific features characterized by (pharmaco)kinetic aspects or by stability of the immunoglobulin
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- C07K2319/00—Fusion polypeptide
- C07K2319/30—Non-immunoglobulin-derived peptide or protein having an immunoglobulin constant or Fc region, or a fragment thereof, attached thereto
Definitions
- GDF15 (Growth/differentiation factor 15) and a fusion polypeptide comprising the Fc region of an immunoglobulin, a pharmaceutical composition comprising the fusion polypeptide, and the in vivo duration of GDF15 including the step of fusing the Fc region of an immunoglobulin It relates to how to increase.
- sustained-release formulations in which a protein or peptide drug is surrounded by a biodegradable polymer matrix is prepared, and the drug is slowly released while the matrix material is slowly decomposed and removed from the body when administered. Has become.
- GDF15 (Growth/differentiation factor 15) is a member of the TGF-beta family, is a 25kDa homodimer, and is a secreted protein circulating in plasma. Plasma levels of GDF15 are related to BMI (Body Mass Index), and GDF15 acts as a long-term regulator of energy homeostasis. GDF15 also protects against pathological conditions such as cardiovascular disease, myocardial hypertrophy, and ischemic injury. In addition, GDF15 plays a role in protection against renal tubular and renal interstitial damage in type 1 diabetes and type 2 diabetes models. In addition, GDF15 has a protective effect against age-related sensory and motor nerve loss, and may contribute to the recovery of peripheral nerve damage. In addition, GDF15 has an effect of reducing body weight and reducing body fat and glucose tolerance, and has an effect of increasing systemic energy consumption and oxidative metabolism. GDF15 exhibits glycemic control effects through weight-dependent and independent mechanisms.
- GDF15 Crowth/differentiation factor 15
- a functional variant thereof with the Fc region of an immunoglobulin to form a fusion polypeptide, compared to the case where it is not fused with the Fc region, the half-life of GDF15 is increased.
- Techniques are provided to enhance the duration of the body, thereby enhancing the pharmacological effect of GDF15 and/or increasing the administration interval.
- One example provides a fusion polypeptide comprising the Fc region of an immunoglobulin with GDF15 or a functional variant thereof.
- the Fc region of the immunoglobulin may be included (linked) at the N-terminus of the GDF15 or a functional variant thereof.
- the fusion polypeptide may include (1) an Fc region of an immunoglobulin and (2) GDF15 or a functional variant thereof in the N-terminal to C-terminal direction.
- the Fc region of the immunoglobulin contained in the fusion polypeptide serves to enhance the stability of GDF15 or a variant thereof, and may be, for example, one having an effect of prolonging half-life (e.g., in vivo half-life).
- the Fc region of the immunoglobulin may be selected from the group consisting of an IgG1 Fc region and an IgG4 Fc region.
- the Fc region of IgG1 may include a CH2 domain, a CH3 domain, or a CH2+CH3 domain of IgG1, and may or may not include a hinge region of IgG1 at the N-terminus.
- the Fc region of IgG4 may include a CH2 domain, a CH3 domain, or a CH2+CH3 domain of IgG4, and may or may not include a hinge region of IgG4 at the N-terminus.
- the fusion polypeptide may further include a peptide linker between the Fc region of the immunoglobulin and GDF15 or a functional variant thereof.
- the peptide linker is preferably a flexible linker so that the Fc region and GDF15 fused to both sides freely function.
- the peptide linker may not be a rigid linker.
- the peptide linker may be a GS linker that repeatedly includes one or more Gly (G) and one or more Ser (S), for example, (GGGGS) n (n is 1 as the number of repetitions of GGGGS (SEQ ID NO: 13)). It may be an integer of to 10 or an integer of 1 to 5 (1, 2, 3, 4, or 5), but is not limited thereto.
- the stability (duration) in the body (or blood) is Characterized by increased (eg, increased half-life in the body or in the blood).
- the immunoglobulin Fc region fused to GDF15 or a functional variant thereof, compared to the immunoglobulin Fc region is not fused to GDF15 or a functional variant thereof, the pharmacological effect (e.g., weight loss effect) is improved. To do.
- fusion polypeptide dimer comprising two of the fusion polypeptides.
- the fusion polypeptide dimer may be a linkage (linkage) between GDF15 or a functional variant thereof of the two fusion polypeptides.
- Another example provides a nucleic acid molecule encoding the fusion polypeptide.
- Another example provides a recombinant vector comprising the nucleic acid molecule.
- Another example provides a recombinant cell comprising the recombinant vector.
- compositions for preventing, improving, alleviating, and/or treating weight loss, diet control (reduction in diet), or metabolic disease.
- Another example is one or more pharmaceutically effective amounts selected from the group consisting of the fusion polypeptide, the fusion polypeptide dimer, the fusion polypeptide encoding nucleic acid molecule, the recombinant vector containing the nucleic acid molecule, and the recombinant cell containing the recombinant vector.
- Prevention, amelioration, alleviation, and/or treatment of metabolic diseases comprising administering to a subject in need thereof, methods of weight loss, diet control (reduction in diet) methods, or prevention, amelioration, alleviation, and/or of metabolic diseases Or provide a treatment method.
- the metabolic disease refers to all diseases caused by metabolic disorders, such as obesity, diabetes (eg, type 2 diabetes), non-alcoholic fatty liver disease (eg, Nonalcoholic steatohepatitis (NASH), etc.). It may be selected from the group consisting of.
- metabolic disorders such as obesity, diabetes (eg, type 2 diabetes), non-alcoholic fatty liver disease (eg, Nonalcoholic steatohepatitis (NASH), etc.). It may be selected from the group consisting of.
- Another example is a method for producing GDF15 or a functional variant thereof having an increased half-life in the body (or blood), comprising the step of expressing the recombinant vector in cells, or GDF15 or a functional variant thereof having an increased half-life in the body (or blood). It provides a method for producing a homodimer comprising a fusion polypeptide or a fusion polypeptide comprising a.
- Another example provides a method of increasing the in vivo duration of GDF15 or a functional variant thereof, comprising fusing (or linking or binding) the Fc region of an immunoglobulin with GDF15 or a functional variant thereof.
- the fusing step may include fusion (or linkage or binding) of the Fc region of the immunoglobulin to the N-terminus of GDF15 or a functional variant thereof with or without a linker.
- GDF15 comprising the step of fusion (or linking or linking) the Fc region of an immunoglobulin with GDF15 or a functional variant thereof through a flexible linker (e.g., GS linker), GDF15, the Fc region of an immunoglobulin, or comprising the same.
- Methods of reducing the immunogenicity of a fusion polypeptide are provided. The step of fusion (or linkage or binding) may be performed in vitro.
- GDF15 is composed of amino acids from 197th (A) to 308th (I) (SEQ ID NO: 1; see Fig. 2; mature form), excluding signal peptide and propeptide, among a total of 308 amino acids (UniProt Q99988):
- GDF15 is the amino acid sequence from the 197th (A) to the 308th (I) of the full-length protein (UniProt Q99988) (see SEQ ID NO: 1, Figure 2; ARNG DHCPLGPGRC CRLHTVRASL EDLGWADWVL SPREVQVTMC IGACPSQFRA) ANMHAQIKTS LHRLKPDTVP APCCVPASYN PMVLIQKTDT GVSLQTYDDL LAKDCHCI) or GDF15 in the range of maintaining the intrinsic activity and structure of the amino acid sequence 80% or more, 85% or more, 90 or more, 95% or more, 96% or more, 97% or more, 98% or more, Or it means a polypeptide essentially containing an amino acid sequence having 99% or more sequence homology.
- the functional variant of GDF15 may be a variant that has been mutated so that GDF15 is more advantageous in forming a dimer structure while maintaining intrinsic activity and structure.
- the functional variant of GDF15 is one or more of the 14 amino acid residues at the N-terminus of the amino acid sequence of GDF15 of SEQ ID NO: 1 (i.e., 14 amino acid residues from the first to the 14th in SEQ ID NO: 1) ( 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, or 14) (e.g., one or more in order from the N terminus), such as all 14 amino acid residues are deleted N-terminal deletion variant (hereinafter, may be denoted as' ⁇ N14GDF15' or'GDF(CRL)').
- the functional variant of GDF15 is the amino acid sequence of SEQ ID NO: 2 (CRLHTVRASL EDLGWADWVL SPREVQVTMC IGACPSQFRA ANMHAQIKTS LHRLKPDTVP APCCVPASYN PMVLIQKTDT GVSLQTYDDL LAKDCHCI in the range of 85% or more maintaining the amino acid sequence and structure of the intrinsic activity of GDF15.
- % Or more, 90 or more, 95% or more, 96% or more, 97% or more, 98% or more, or 99% or more may be a polypeptide essentially containing an amino acid sequence having sequence homology.
- the Fc of immunoglobulin fused with GDF15 or a functional variant thereof serves to enhance the stability of the GDF15 or a functional variant thereof, such as extending half-life (eg, in vivo half-life), and / or reducing renal filtration effects. It may be to have.
- the Fc region of the immunoglobulin may be selected from IgG1 Fc and IgG4 Fc.
- the Fc region of IgG1 may include a CH2 domain, a CH3 domain, or a CH2+CH3 domain of IgG1, and may or may not include a hinge region of IgG1 at the N-terminus.
- the Fc region of IgG4 may include a CH2 domain, a CH3 domain, or a CH2+CH3 domain of IgG4, and may or may not include a hinge region of IgG4 at the N-terminus.
- IgG1 may be derived from primates such as humans or rodents such as mice and rats, and may be, for example, human IgG1 (UniProtKB P01857).
- IgG4 may be derived from a primate such as human or rodent such as mouse or rat, and may be, for example, human IgG4 (UniprotKB P01861).
- the Fc region of IgG1 may include a CH2 domain, a CH3 domain, or a CH2+CH3 domain of IgG1, and may or may not include a hinge region of IgG1 at the N-terminus.
- the Fc region of IgG4 may include a CH2 domain, a CH3 domain, or a CH2+CH3 domain of IgG4, and may or may not include a hinge region of IgG4 at the N-terminus.
- the Fc of IgG1 is a polypeptide containing a CH2 domain and a CH3 domain of human IgG1 (SEQ ID NO: 3), or a hinge region of human IgG1 at the N-terminus of the amino acid sequence of SEQ ID NO: 3 (SEQ ID NO: 4 ) May be a polypeptide further comprising.
- the Fc region of IgG1 is of the above SEQ ID NO: 3 or'N-terminal-(SEQ ID NO: 4)-(SEQ ID NO: 3) -C terminal' in a range that maintains the intrinsic activity and structure of IgG1 Fc. It can be interpreted as including an amino acid sequence having a sequence homology of 80% or more, 85% or more, 90 or more, 95% or more, 96% or more, 97% or more, 98% or more, or 99% or more of an amino acid sequence.
- the Fc of IgG4 further comprises a polypeptide containing a CH2 domain and a CH3 domain of human IgG4 (SEQ ID NO: 5) or a hinge region of human IgG4 (SEQ ID NO: 10) at the N-terminus of the amino acid sequence of SEQ ID NO: 5 It may be a polypeptide.
- the Fc region of IgG4 is SEQ ID NO: 5 or'N-terminal-(SEQ ID NO: 10)-(SEQ ID NO: 5)-C-terminal' in a range that maintains the intrinsic activity and structure of IgG4 Fc.
- amino acid sequence or the amino acid sequence of a variant Fc of IgG4 described below
- the human IgG4 Fc region is advantageous because the binding power between Fc ⁇ R and complement factors is low compared to other IgG sub-types.
- the human IgG4 Fc region may have an effector function further reduced by including amino acid substitutions.
- the amino acid substitution of the human IgG4 Fc region to reduce the effector function is the substitution of phenylalanine, which is the 234th residue of human IgG4 (UniprotKB P01861), to alanine and the 235th residue leucine to alanine (CH2-CH3 domain of IgG4 Fc. Included in the site;
- the amino acid residue number is according to EU numbering [Kabat, EA et al. (1991) Sequences of Proteins of Immunological Interest, 5th edition, US Dept. of Health and Human Services, Bethesda, MD, NIH Publication no. 91 -3242]).
- the Fc region of the IgG4 may contain amino acid substitutions that stabilize the formation of heavy chain dimers and prevent the formation of a half-IgG4 Fc chain.
- amino acid substitutions may include substitution of proline for the 228th amino acid residue serine (according to EU numbering; contained in the hinge region) of the Fc region of human IgG4 (UniprotKB P01861).
- the Fc region in order to reduce the immunogenicity (Immunogenicity), may not include a mutation other than the above mutation, but is not limited thereto.
- the Fc region of the immunoglobulin is a human IgG4 Fc CH2-CH3 domain (SEQ ID NO: 5 (general formula); SEQ ID NO: 6 (wild type), or SEQ ID NO: 7, 8, or 9 (variant)) Or a hinge region of human IgG4 (SEQ ID NO: 10 (general formula); SEQ ID NO: 11 (wild type), or SEQ ID NO: 12 (variant type)) at the N-terminus of the CH2-CH3 domain. .
- the fusion polypeptide provided herein includes the Fc region of an immunoglobulin and GDF15 or a functional variant thereof linked to the C-terminus of the Fc region of the immunoglobulin.
- the Fc region of the immunoglobulin and GDF15 or a functional variant thereof are as described above.
- the Fc region of the immunoglobulin and GDF15 or a functional variant thereof may be covalently or non-covalently linked, through an appropriate linker (eg, a peptide linker), or directly linked without a linker.
- the peptide linker may be a polypeptide consisting of 1 to 20, 1 to 15, 1 to 10, 2 to 20, 2 to 15, or 2 to 10 arbitrary amino acids, and the There is no limitation on the type of amino acid contained.
- the peptide linker is preferably a flexible linker in order to allow the Fc region fused to both sides (stabilizing activity such as increasing half-life, reducing renal filtration, etc.) and GDF15 to freely function.
- the peptide linker may not be a rigid linker.
- the peptide linker may include, for example, one or more amino acid residues selected from the group consisting of Gly, Asn, Ser, Glu, and Lys, and may include neutral amino acids such as Thr and/or Ala, Without being limited thereto, amino acid sequences suitable for peptide linkers are known in the art.
- the peptide linker may be a GS linker repeatedly containing one or more Gly (G) and one or more Ser (S), for example, (GGGGS) n (n is the number of repetitions of GGGGS (SEQ ID NO: 13)) May be an integer of 1 to 10 or an integer of 1 to 5 (1, 2, 3, 4, or 5), but is not limited thereto.
- Gly (glycine) is an amino acid whose R group is -H and is nonpolar, but has excellent mobility due to its high degree of freedom (phi, psi angle)
- Ser (serine) is an amino acid whose R group is -CH2-OH.
- the size is small and polar, it is advantageous in maintaining the stability of the linker by forming a hydrogen bond with water, thereby contributing to reducing possible non-specific interactions between the GS linker and the GDF15 or Fc region.
- the GS linker since the GS linker has a flexible structure, it has an advantage of low immunogenicity.
- the peptide linker included in the fusion polypeptide proposed herein does not include amino acid residues other than the above-described GS linker [(GGGGS)n (n is an integer of 1 to 10 or an integer of 1 to 5)]. It may not be.
- the fusion polypeptide may further include a peptide linker between the Fc region of the immunoglobulin and GDF15 or a functional variant thereof.
- the peptide linker may be a GS linker repeatedly containing one or more Gly (G) and one or more Ser (S), for example, (GGGGS) n (n is the number of repetitions of GGGGS (SEQ ID NO: 13)) May be an integer of 1 to 10 or an integer of 1 to 5 (1, 2, 3, 4, or 5), but is not limited thereto.
- the fusion polypeptide can be produced recombinantly or chemically synthesized.
- the fusion polypeptide is encoded by one reading frame in which expression is regulated by one transcription initiation regulator (eg, a promoter, etc.) (expressed as "single strand" ) Can be.
- one transcription initiation regulator eg, a promoter, etc.
- GDF15 or a functional variant thereof fused with an Fc region of an immunoglobulin is GDF15 or a functional variant thereof in which the Fc region of an immunoglobulin is not fused, and/or an Fc region and GDF15 other than the GS linker described above.
- stability in the body (or blood) is increased (eg, increased half-life in blood or in the body), and/or immunogenicity is decreased.
- the immunoglobulin Fc region fused to GDF15 or a functional variant thereof compared to the immunoglobulin Fc region is not fused to GDF15 or a functional variant thereof, the pharmacological effect (e.g., weight loss effect) is improved. To do.
- the functionally active form of GDF15 is the homodimer form.
- a fusion polypeptide dimer in which two fusion polypeptides (a first fusion polypeptide and a second fusion polypeptide) are bound.
- the first fusion polypeptide comprises an Fc region of a first immunoglobulin and a first GDF15 or a functional variant thereof
- the second fusion polypeptide comprises an Fc region of a second immunoglobulin and a second GDF15 or a functional variant thereof.
- the first fusion polypeptide and the second fusion polypeptide may be a first GDF15 or a functional variant thereof and a second GDF15 or a functional variant thereof linked through a covalent bond (e.g., a disulfide bond).
- the Fc region of the first immunoglobulin and the Fc region of the second immunoglobulin are covalently bonded (eg, disulfide bonds, etc.) or non-covalent bonds (eg, knobs & holes, electrostatic interactions). Action, hydrophobic interaction, etc.) may be connected to each other (see FIG. 1).
- the Fc region of the immunoglobulin and GDF15 or a functional variant thereof are as described above, and the Fc region of the first immunoglobulin and the Fc region of the second immunoglobulin, and the first GDF15 or a functional variant thereof and the second GDF15 or a functional variant thereof Each of the variants can be the same or different.
- the first fusion polypeptide and the second fusion polypeptide are each in the form of a single strand (chain; for example, a polypeptide encoded by one reading frame), and the Fc region of the first immunoglobulin and the Fc of the second immunoglobulin A structure in which at least one of the region, and the first GDF15 or functional variant thereof and the second GDF15 or functional variant thereof, and the first fusion polypeptide and the second fusion polypeptide formed by linking them is in a dimeric form (e.g., first or When the Fc region of the first or second immunoglobulin included in the second fusion polypeptide is in a form in which two strands (two molecules) are linked by a disulfide bond or the like)
- the fusion partner ie, the Fc region of an immunoglobulin
- the fusion partner is preferably linked to the N-terminus of GDF15.
- GDF15 binds to its receptor, GFRAL (GDNF family receptor alpha-like; for example, GenBank Accession no. NP_997293.2) to form a GDF15-GFRAL complex structure.
- GFRAL GDNF family receptor alpha-like; for example, GenBank Accession no. NP_997293.2
- the GDF15 monomer and the GFRAL monomer are each bound, and the N-terminus of GDF15 is located in the middle of the GDF15 dimer.
- the N-terminal four amino acid residues that can be fused to GDF15 are not observed in the X-ray structure, so it is predicted that they do not have a specific structure.
- the 14 amino acid residues at the N-terminus of GDF15 are fixed with a Cys7-Cys14 disulfide bond and are not involved in the interaction between monomers in the GDF15 dimer.
- the 14 amino acid residues at the N-terminus of GDF15 are structurally located at a distance that cannot interact (bind) with the GFRAL receptor, so even if the N-terminal 14 amino acid residues of GDF15 are deleted, GDF15 binds to the GFRAL receptor. No impact is expected.
- the GDF15 monomer has four disulfide bonds in the molecule and one disulfide bond between the dimer molecule.
- the disulfide bonds in the molecule are believed to play a role in stabilizing the structure, and the disulfide bond connecting Cys7-Cys14 is located at the edge of the monomer structure and serves to fix the N-terminal loop structure.
- N-terminal 14 amino acid residues do not play a direct role in binding to the GFRAL receptor, so removing one or more of them does not affect the function and structure of GDF15, but Cys15 forms a disulfide bond with Cys88, so Cys15 cannot be removed. In this case, it may affect the three-dimensional structure of GDF15.
- the region that can be removed while maintaining the function and structure of GDF15 is up to 14 amino acids at the N-terminus (i.e., in SEQ ID NO: 1, at least one of the 14 amino acids from Ala1 to Cys14, for example, in order from the N-terminus One or more (1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, or 14)).
- the GDF15 monomer may be a wild type (SEQ ID NO: 1) or a 14 amino acid deletion form at the N-terminus in the wild type ( ⁇ N14GDF15; SEQ ID NO: 2).
- Fc- ⁇ N14GDF15 or Fc-wtGDF15 and GFRAL are functionally in dimer form, and in the case of Fc- ⁇ N14GDF15, the formation of Fc dimer and ⁇ N14GDF15 dimer, respectively, has a structural arrangement that facilitates formation. Therefore, Fc- ⁇ N14GDF15 may be more advantageous in terms of dimer formation than Fc-wtGDF15 without interfering with the formation of a complex with GFRAL.
- the in vivo (blood) half-life in mammals of GDF15 or a functional variant thereof contained in the fusion polypeptide or fusion polypeptide dimer provided herein is compared to GDF15 or a functional variant thereof in which the Fc region of an immunoglobulin is not fused. , About 1.5 times or more, about 2 times or more, about 2.5 times or more, about 3 times or more, about 3.5 times or more, about 4 times or more, about 4.5 times or more, about 5 times or more, about 5.5 times or more, about 6 times or more , About 7 times or more, about 8 times or more, about 9 times or more, or about 10 times or more.
- GDF15 in the form of a fusion polypeptide to which the Fc region of an immunoglobulin is bound, or a functional variant thereof is GDF15 in a form in which the Fc region of an immunoglobulin is not linked, or a functional variant thereof.
- the administration interval can be lengthened.
- the fusion polypeptide comprising the Fc region of GDF15 or a functional variant thereof and an immunoglobulin may be produced by a conventional chemical synthesis method or a recombinant method, and may not be naturally occurring.
- the term "vector” refers to an expression means for expressing a gene of interest in a host cell, for example, a plasmid vector, a cozmid vector, and a bacteriophage vector, an adenovirus vector, a retroviral vector, and It may be selected from the group consisting of viral vectors such as adeno-associated viral vectors.
- the vector that can be used in the recombinant vector is a plasmid (e.g., pcDNA series, pSC101, pGV1106, pACYC177, ColE1, pKT230, pME290, pBR322, pUC8/9, pUC6, pBD9, pHC79, pIJ61, pLAFR1, pHV14, pGEX series, pET series, pUC19, etc.), phage (e.g., ⁇ gt4 ⁇ B, ⁇ -Charon, ⁇ z1, M13, etc.) or virus (e.g., SV40, etc.), but is not limited thereto.
- plasmid e.g., pcDNA series, pSC101, pGV1106, pACYC177, ColE1, pKT230, pME290, pBR322, pUC8/9, pUC6, pBD9, pHC79, pIJ
- the nucleic acid molecule encoding the fusion polypeptide may be operably linked to a promoter.
- operatively linked refers to a functional linkage between a nucleic acid expression control sequence (eg, a promoter sequence) and another nucleic acid sequence.
- the regulatory sequences can be “operatively linked” to control transcription and/or translation of other nucleic acid sequences.
- the recombinant vector can be typically constructed as a vector for cloning or an expression vector for expression.
- the expression vector may be a conventional one used in the art to express foreign proteins in plants, animals or microorganisms.
- the recombinant vector can be constructed through various methods known in the art.
- the recombinant vector can be expressed using eukaryotic cells as a host.
- the recombinant vector includes the nucleic acid molecule to be expressed, the promoter described above, the ribosome binding site, the secretion signal sequence (refer to Publication No. 2015-0125402), and/or transcription/
- the origin of replication operating in eukaryotic cells may include the f1 origin of replication, SV40 origin of replication, pMB1 origin of replication, adeno origin of replication, AAV origin of replication, and/or BBV origin of replication, but are limited thereto. no.
- a promoter derived from the genome of a mammalian cell e.g., metallotionine promoter
- a promoter derived from mammalian virus e.g., adenovirus late promoter, vaccinia virus 7.5K promoter, SV40 promoter, A cytomegalovirus promoter, a fusion promoter (KR 10-1038126 or KR 10-1868139), or the tk promoter of HSV
- a promoter derived from the genome of a mammalian cell e.g., metallotionine promoter
- a promoter derived from mammalian virus e.g., adenovirus late promoter, vaccinia virus 7.5K promoter, SV40 promoter, A cytomegalovirus promoter, a fusion promoter (KR 10-1038126 or KR 10-1868139), or the tk promoter of HSV
- all commonly available secretion signal sequences can be used as the secreti
- the recombinant cell may be obtained by introducing (transforming or transfecting) the recombinant vector into an appropriate host cell.
- the host cell may be selected from all eukaryotic cells capable of stably and continuously cloning or expressing the recombinant vector.
- Eukaryotic cells usable as a host include yeast ( Saccharomyces cerevisiae ), insect cells, plant cells, and animal cells, and the like, for example, mice (e.g., COP, L, C127, Sp2/0, NS-0, NS-1, At20, or NIH3T3), rat (e.g., PC12, PC12h, GH3, or MtT), hamster (e.g., BHK, CHO, GS gene defect CHO, or DHFR gene defect CHO), monkey (eg, COS ( COS1, COS3, COS7, etc.), CV1 or Vero), human (e.g., HeLa, HEK-293, retinal-derived PER-C6, cells derived from diploid fibroblasts, myeloma cells or HepG2), other animal cells (e.g. , MDCK, etc.), insect cells (eg, Sf9 cells, Sf21 cells, Tn-368 cells, BTI-TN
- Polypeptide dimers can be prepared.
- the method for preparing the fusion polypeptide or the fusion polypeptide dimer may include culturing a recombinant cell containing the nucleic acid molecule. The step of culturing may be performed under conventional culture conditions. In addition, the manufacturing method may further include separating and/or purifying the fusion polypeptide or fusion polypeptide dimer from the culture after the culturing step.
- Transport (introduction) of the nucleic acid molecule or a recombinant vector containing the same into a host cell may use a transport method well known in the art.
- a transport method well known in the art.
- the host cell is a eukaryotic cell
- a microinjection method, a calcium phosphate precipitation method, an electroporation method, a liposome-mediated transfection method, and gene bombadment may be used, but are not limited thereto. .
- the method of selecting the transformed (recombinant vector introduced) host cells can be easily carried out according to a method well known in the art using a phenotype expressed by a selection label.
- the selection marker is a specific antibiotic resistance gene, it is possible to easily select a recombinant cell into which the preparation vector has been introduced by culturing in a medium containing the antibiotic.
- At least one pharmaceutically effective amount selected from the group consisting of the fusion polypeptide, the fusion polypeptide dimer, the fusion polypeptide encoding nucleic acid molecule, the recombinant vector containing the nucleic acid molecule, and the recombinant cell containing the recombinant vector is used for metabolic disease.
- Methods of weight loss, diet control (reduction in diet), or methods of preventing, ameliorating, alleviating, and/or treating metabolic diseases, comprising administering to a subject in need of prevention, improvement, alleviation, and/or treatment. is provided.
- the metabolic disease refers to all diseases caused by metabolic disorders, such as obesity, diabetes (eg, type 2 diabetes), non-alcoholic fatty liver disease (eg, Nonalcoholic steatohepatitis (NASH), etc.). It may be selected from the group consisting of.
- metabolic disorders such as obesity, diabetes (eg, type 2 diabetes), non-alcoholic fatty liver disease (eg, Nonalcoholic steatohepatitis (NASH), etc.). It may be selected from the group consisting of.
- a pharmaceutically effective amount refers to the content or dosage of an active ingredient capable of obtaining a desired effect.
- the active ingredient in the pharmaceutical composition (GDF15 or a functional variant thereof and a fusion polypeptide containing the Fc region of an immunoglobulin, a fusion polypeptide dimer, a fusion polypeptide encoding nucleic acid molecule, a recombinant vector containing the nucleic acid molecule, and the recombinant
- the content or dosage of one or more selected from the group consisting of recombinant cells containing the vector) is the formulation method, the mode of administration, the patient's age, weight, sex, pathological condition, food, administration time, administration interval, administration route, excretion.
- one dose of the active ingredient is 0.001 to 1000 mg/kg, 0.01 to 100 mg/kg, 0.01 to 50 mg/kg, 0.01 to 20 mg/kg, 0.01 to 10 mg/kg, 0.01 to 5 mg/kg , 0.1 to 100 mg/kg, 0.1 to 50 mg/kg, 0.1 to 20 mg/kg, 0.1 to 10 mg/kg, 0.1 to 5 mg/kg, 1 to 100 mg/kg, 1 to 50 mg/kg, 1 to It may be in the range of 20 mg/kg, 1 to 10 mg/kg, or 1 to 5 mg/kg, but is not limited thereto.
- the content of the active ingredient in the pharmaceutical composition is, based on the weight of the total pharmaceutical composition, 0.01% to 99.9% by weight, 0.01% to 90% by weight, 0.01% to 80% by weight, 0.01% to 70% by weight %, 0.01% to 60%, 0.01% to 50%, 0.01% to 40%, 0.01% to 30%, 1% to 99.9%, 1% to 90%, 1% to 80%, 1% to 70%, 1% to 60%, 1% to 50%, 1% to 40%, 1% to 30%, 5% % To 99.9%, 5% to 90%, 5% to 80%, 5% to 70%, 5% to 60%, 5% to 50%, 5% to 40 wt%, 5 wt% to 30 wt%, 10 wt% to 99.9 wt%, 10 wt% to 90 wt%, 10 wt% to 80 wt%, 10 wt% to 70 wt%, 10 wt% to 60 wt% %, 10% to 50% by
- the administration interval (temporal interval between two adjacent administrations) of the active ingredient provided herein or a pharmaceutical composition containing the same can be adjusted according to the concentration or state of the active ingredient (whether or not there is a mutation), or the condition or symptom of the patient, For example, 2 days or more, 3 days or more, 4 days or more, 5 days or more, 6 days or more, 7 days or more, 8 days or more, 9 days or more, 10 days or more, 2 weeks or more, 3 weeks or more, 4 weeks or more, It can be at least 6 weeks, at least 8 weeks, at least 10 weeks, or at least 12 weeks.
- the maximum administration interval may be about 2 weeks, about 3 weeks, about 4 weeks, about 1 month, about 2 months, or about 3 months, but is not limited thereto, and the concentration or state of the active ingredient (such as mutation), Or it can be increased or decreased according to the patient's condition or symptoms. In one embodiment, the administration interval may be appropriately prescribed within about 1 week to about 3 months.
- the pharmaceutical composition may further include a pharmaceutically acceptable carrier in addition to the active ingredient.
- the carrier is commonly used in the formulation of proteins, nucleic acids, or drugs containing cells, and is lactose, dextrose, sucrose, sorbitol, mannitol, starch, gum acacia, calcium phosphate, alginate, gelatin, calcium silicate, It may be one or more selected from the group consisting of microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methyl cellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, mineral oil, etc. However, it is not limited thereto.
- the pharmaceutical composition may further include one or more selected from the group consisting of diluents, excipients, lubricants, wetting agents, sweetening agents, flavoring agents, emulsifying agents, suspending agents, preservatives, and the like, which are commonly used in the manufacture of pharmaceutical compositions.
- the administration target of the pharmaceutical composition may be a mammal, including a human, a primate including a monkey, a rodent including a mouse, or a rat, or a cell, tissue, cell culture or tissue culture derived therefrom.
- the pharmaceutical composition may be administered by oral administration or parenteral administration, or may be administered by contacting cells, tissues, or body fluids.
- parenteral administration may be administered by subcutaneous injection, intramuscular injection, intravenous injection, intraperitoneal injection, endothelial administration, topical administration, intranasal administration, intrapulmonary administration, and rectal administration.
- the oral composition should be coated with an active agent or formulated to protect it from degradation in the stomach.
- the pharmaceutical composition may be in the form of a solution, suspension, syrup, or emulsion in an oil or aqueous medium, or may be formulated in the form of extracts, powders, granules, tablets or capsules, and a dispersant or a stabilizer is additionally added for formulation.
- Can include.
- Another example provides a method of increasing the in vivo duration of GDF15 or a functional variant thereof, comprising fusing (or linking or binding) the Fc region of an immunoglobulin with GDF15 or a functional variant thereof.
- the fusing step may include fusion (or linkage or binding) of the Fc region of the immunoglobulin to the N-terminus of GDF15 or a functional variant thereof with or without a linker.
- Another example is the immunogenicity of GDF15, the Fc region of an immunoglobulin, or a fusion polypeptide comprising the same, comprising the step of fusion (or linking or linking) the Fc region of an immunoglobulin with GDF15 or a functional variant thereof through a flexible linker.
- the step of fusion (or linkage or binding) may be performed in vitro.
- GDF15 or a functional variant thereof fused with the Fc region of immunoglobulin provided herein compared to the case where it is not fused with the Fc region of immunoglobulin, has a longer duration when administered in vivo, so the administration interval can be extended and administered through it. Since the dose can be reduced, it has an advantageous effect in terms of administration convenience and/or economy, and has excellent pharmacological effects, and thus can be usefully applied to fields requiring treatment of GDF15 or a functional variant thereof.
- FIG. 1 is a schematic diagram of a fusion polypeptide according to an embodiment.
- Figure 2 shows the amino acid sequence of GDF15 (wild type; mature form; SEQ ID NO: 1) (N-terminal to C-terminal direction).
- 3A and 3B show a complex structure in which Fc- ⁇ N14GDF15 (3a) or Fc-wtGDF15 (3b) and GFRAL are bound.
- FIG. 4 is a schematic diagram schematically showing a fusion polypeptide gene according to an embodiment.
- FIG. 5 is a graph showing changes in body weight of the IgG1-GDF (CRL) administration group and the HIgG1-GDF (CRL) administration group.
- FIG. 6 is a graph showing the rate of body weight change on the 7th day after administration of the IgG1-GDF (CRL) administration group and the HIgG1-GDF (CRL) administration group.
- FIG. 7 is a graph showing changes in body weight of the IgG4-GDF15 administration group and the IgG4-GDF (CRL) administration group.
- FIG 8 is a graph showing the rate of change in body weight on days 7 (blue bars) and 14 days (red bars) of the IgG4-GDF15 administration group and the IgG4-GDF (CRL) administration group.
- FIG 9 is a graph showing the cumulative feed intake up to the 7th day after administration of the IgG1-GDF (CRL) administration group and the HIgG1-GDF (CRL) administration group.
- Figure 10 is a graph showing the cumulative feed intake up to the 7th, 14th, and 19th days after administration of the IgG4-GDF15 administration group and the IgG4-GDF (CRL) administration group.
- FIG. 11 is a graph showing changes in the concentration of the fusion polypeptide in blood (in serum) according to the elapsed time after administration of the fusion polypeptide.
- Immunoglobulin Fc with or without Hinge (IgG1-Fc (without hinge: SEQ ID NO: 3; HIgG1-Fc (with hinge; [SEQ ID NO: 4]-[SEQ ID NO: 3]), Mutated IgG4-Fc (without hinge; SEQ ID NO: 7) or Mutated HIgG4-Fc (including hinge: [SEQ ID NO: 12]-[SEQ ID NO: 7]) is the target polypeptide Mature GDF15 (shown as GDF or GDF15; SEQ ID NO: 1), or GDF15 variant (GDF ( CRL); N-terminal 14 amino acid deletion: fusion polypeptide fused with SEQ ID NO: 2) IgG1-GDF (CRL), HIgG1-GDF (CRL), IgG4-GDF, HIgG4-GDF, IgG4-GDF (CRL) , And HIgG4-GDF(CRL) (see Fig. 1.) The amino
- IgG4-GDF15 (without hinge) & HIgG4-GDF15 (with hinge) (N-terminal to C-terminal direction) Amino acid sequence Sequence number Signal Peptide MHRPEAMLLL LTLALLGGPT WA 17 Mutated IgG4 Fc Hinge ESKYGPPCPP CP 12 CH2-CH3 APEAAGGPSV FLFPPKPKDT LMISRTPEVT CVVVDVSQED PEVQFNWYVD GVEVHNAKTK PREEQFNSTY RVVSVLTVLH QDWLNGKEYK CKVSNKGLPS SIEKTISKAK GQPREPQVYT LPPSQEEPEMTK YGFKVSCLVNGDS 7 Linker GGGGSGGGGS GGGGSGGGGS 18 GDF15 ARNGDHCPLG PGRCCRLHTV RASLEDLGWA DWVLSPREVQ VTMCIGACPS QFRAANMHAQ IKTSLHRLKP DTVPAPCCVP ASYNPMVLIQ
- a gene encoding mature GDF15 (SEQ ID NO: 14) was synthesized in Bioneer with reference to the amino acid of UniprotKB Q99968.
- the gene coding for human IgG1 Fc including Hinge or the gene coding for human IgG1 Fc not including Hinge was obtained by PCR using a plasmid containing a core Hinge of human IgG1 and a gene coding for IgG1 Fc.
- the gene coding for human IgG4 Fc including Hinge or the gene coding for human IgG4 Fc not including Hinge was obtained through PCR using a plasmid containing genes coding for Hinge and IgG4 Fc of human IgG4.
- pDHDD-D1G1 a variant of pcDNA3.1(+) (Invitrogen, Cat.No.V790-20) (including the promoter of KR10-1868139B1) was cut with BamHI and NotI, and the genes (mature GDF15, IgG1-Fc, IgG4-Fc) were combined to insert a gene having the following structure (see FIG. 4) to prepare each recombinant vector.
- the prepared recombinant expression vectors pHIgG1-GDF (CRL), pIgG1-GDF (CRL), pHIgG4-GDF (CRL), pIgG4-GDF (CRL), pHIgG4-GDF and pIgG4-GDF were used in ExpiCHO-S TM cells (Thermo Fisher Scientific ) And cultured in ExpiCHO Expression Medium (Thermo Fisher Scientific; 400 mL) for 12 days (Fed-Batch Culture; Day 1 & Day 5 Feeding), the fusion polypeptide HIgG1-GDF (CRL), IgG1-GDF ( CRL), HIgG4-GDF (CRL), IgG4-GDF (CRL), HIgG4-GDF and IgG4-GDF were expressed.
- the fusion polypeptide was purified from the cell culture prepared in Example 1.1 using Protein A Affinity Chromatography.
- the culture solution of the fusion polypeptide from which the cells were removed was filtered through a 0.22 ⁇ m filter.
- a column packed with MabSelect SuRe TM pcc (GE Healthcare Life Sciences) resin was mounted on AKTA TM Pure (GE Healthcare Life Sciences), and Phosphate Buffered Saline (PBS, 10 mM Sodium Phosphate, 150 mM NaCl, pH 7.4) was poured.
- PBS Phosphate Buffered Saline
- the given column was equilibrated.
- PBS was again flowed to wash the column.
- an elution buffer (0.1 M Sodium Citrate pH 3.5) was flowed through the column to elute the target fusion polypeptide.
- the eluate was immediately adjusted to neutral pH by adding 1 M Tris pH 8.5.
- fractions with high concentration of fusion polypeptide and high purity were collected and stored frozen.
- the eluted fraction sample containing the fusion polypeptide was concentrated with PBS or 20 mM Tris pH 8.0, 150 mM NaCl, and buffer exchange was performed. Implemented.
- Quantitative analysis of the fusion polypeptide was performed by measuring the absorbance of 280 nm and 340 nm in a UV Spectrophotometer (G113A, Agilent Technologies), and calculating the protein concentration through the following calculation formula.
- the extinction coefficient of each material was theoretically calculated using the amino acid sequence (Table 5).
- Extinction coefficient of fusion polypeptide Sample name Extinction coefficient (0.1%, 1 mg/mL) IgG1-GDF (CRL) 1.317 HIgG1-GDF (CRL) 1.22 IgG4-GDF (CRL) 1.334 IgG4-GDF 1.287
- Example 1 The pharmacological effect of the fusion polypeptide produced and purified in Example 1 was tested in mice (C57BL/6J, 6 weeks old, male, 100 mice; Raon Bio Co., Ltd.).
- the DIO mouse model is an animal model widely used for evaluating the efficacy of improving diabetes and insulin resistance because it exhibits clinical characteristics of type 2 diabetes such as hyperlipemia, insulin resistance, and hyperglycemia. Since a large amount of comparable basic data has been accumulated, this model was selected as appropriate for the pharmacological effect test of this example.
- the mouse model fed the obesity feed for the above 8 weeks went through a two-week quarantine and acclimatization period, and during this period, general symptoms were observed once a day to check whether they were healthy and suitable for the conduct of the experiment, and healthy animals were selected. .
- the tail of the animal was marked with a red oil pen at the time of receipt, and a temporary individual identification card (test name, subject number, arrival time) was attached to the breeding box during the quarantine acclimatization period.
- a temporary individual identification card test name, subject number, arrival time
- an individual was marked using a black oil pen on the tail of the animal, and an individual identification card (test name, group information, individual number, sex, time of arrival, administration period) was attached to each cage.
- test substance fusion polypeptide
- sterilized distilled physiological saline was administered subcutaneously to all animals at 200 uL/head from 3 days before administration of the test substance using a 1 mL syringe. Pre-adaptation training for subcutaneous administration was performed.
- Body weight and feed intake were measured, and group separation was performed so that the average of the two measurements between groups was similar based on body weight.
- Administration of the test substance was started from the day after group separation. Remaining animals that were not selected were excluded from the test system after group separation.
- the information of the high fat feed (obesity feed; High fat diet (HFD)) fed to the C57BL/6J-DIO is as follows:
- the feed was fed by free feeding (purification and feeding during the test period).
- water was filtered with a filter oil-water sterilizer, irradiated with ultraviolet rays, and freely ingested using a drinking water bottle made of polycarbonate (250 mL).
- test substances HgG1-GDF(CRL) and IgG1-GDF(CRL)
- control substance GDF15 R&D Systems
- the dose amount was 5 mL/kg, and the dose amount for each individual was calculated based on the recently measured body weight, and administered subcutaneously once the day of the start of the test using a disposable syringe (1 mL).
- the test substance was administered only once on the start of the test.
- a control group administered with the control substance GDF15 was prepared, and in the case of the control group administered with GDF15, a total of 5 administrations were performed once a day for 5 days, and all administrations were performed from 9 am.
- test group composition and dosage are summarized in Table 6 below:
- IgG1-GDF fusion polypeptide administration group High Fat Diet Test substance Route of administration Dosage (nmol/kg) Dosing volume (mL/kg) Number of animals X Vehicle Subcutaneous - 5 5 O Vehicle, qd Subcutaneous - 5 5 O Vehicle, qw Subcutaneous - 5 5 O GDF15 Subcutaneous 2.86 5 3 O IgG1-GDF (CRL) Subcutaneous One 5 5 O IgG1-GDF (CRL) Subcutaneous 10 5 5 O HIgG1-GDF (CRL) Subcutaneous One 5 5 O HIgG1-GDF (CRL) Subcutaneous 10 5 5 5
- the observation, measurement, and test schedule for the test group was set as Day 0, and 7 days from the start date of administration as one week.
- the body weight of each mouse was measured on the start date of administration of the test substance (before administration), and the body weight was measured every day thereafter (up to 9 days). The amount of test substance administered was determined based on the most recently measured body weight.
- feed intake was measured every day, and the amount of feeding was measured using an electronic scale for each breeding box, and then the remaining amount was measured to calculate the daily feed intake. Individuals who gnaw food severely were excluded from the measurement.
- Both the test substance (IgG4-GDF and IgG4-GDF (CRL)) and the control substance Semaglutide (Bachem) were administered subcutaneously.
- the dose amount was 5 mL/kg, and the dose amount for each individual was calculated based on the recently measured body weight, and administered subcutaneously once the day of the start of the test using a disposable syringe (1 mL).
- the test substance was administered only once on the start of the test, and for comparison, a control group administered with the control substance Semaglutide was prepared, and in the case of the control group administered with Semaglutide, it was administered once a day, and all administrations proceeded from 9 am. .
- test group composition and dosage are summarized in Table 8 below:
- composition of IgG4-GDF15 fusion polypeptide administration group High Fat Diet Test substance Route of administration Dosage (nmol/kg) Dosing volume (mL/kg) Number of animals O Vehicle, qw Subcutaneous - 5 5 O Semaglutide, qd Subcutaneous 3 5 5 O IgG4-GDF Subcutaneous One 5 5 O IgG4-GDF Subcutaneous 10 5 5 O IgG4-GDF (CRL) Subcutaneous One 5 5 O IgG4-GDF (CRL) Subcutaneous 10 5 5 5
- the body weight of each mouse was measured on the start day of administration of the test substance (before administration), and the body weight was measured every day thereafter (up to 19 days).
- the amount of test substance administered was determined based on the most recently measured body weight.
- feed intake was measured every day, and the amount of feeding was measured using an electronic scale for each breeding box, and then the remaining amount was measured to calculate the daily feed intake. Individuals who gnaw food severely were excluded from the measurement.
- Example 2.1.1 The change in weight measured in Example 2.1.1 is shown in FIGS. 5 and 6, and in Table 9 (Body Weight (Group,% of initial)).
- FIG. 5 shows fusion protein IgG1-GDF (CRL) The body weight change at one time of administration was compared with the negative control group (vehicle administration group) and the positive control group (GDF15 daily administration group).
- FIG. 6 is a graph showing the results of the 7th day extracted from the results of FIG. 5.
- Example 2.1.2 The weight change measured in Example 2.1.2 is shown in Fig. 7 and Table 10 (Body Weight (Group,% of initial)).
- FIG. 7 and 8 and Table 10 show fusion proteins IgG4-GDF and IgG4-GDF ( CRL) is shown in comparison with the negative control group (vehicle administration group) and the positive control group (Semaglutide daily administration group).
- FIG. 8 is a graph showing the results of the 7th and 14th days extracted from the results of FIG. 7.
- the weight loss effect at the same dose was compared to the GDF (CRL) fusion polypeptide with Hinge-containing IgG4 Fc (Mutated). It was found to be excellent.
- the weight loss effect of this IgG4 Fc fusion polypeptide can be said to be comparable when Semaglutide is administered once a day throughout the test period.
- Example 2.1.1 The changes in the feed intake measured in Example 2.1.1 are shown in Table 11 and Fig. 9 (cumulative intake up to day 6), respectively.
- Table 12 and FIG. 10 (cumulative intake amount up to the 7th day, the 14th day and the 19th day) are shown for the change in feed intake measured in Example 2.1.2.
- GDF15 or GDF (CRL) is a fusion polypeptide fused with an IgG4 Fc (Mutated) that does not contain Hinge, compared with 10 nmol/kg administration groups, in the case of the fusion polypeptide administration group with full-length GDF15 fusion, the dietary inhibitory ability is about 10 days. In the case of the GDF (CRL) fused fusion polypeptide administration group, the dietary inhibitory ability lasted about 2 weeks. That is, compared to the full-length GDF15 fusion polypeptide, the GDF (CRL) fusion polypeptide was found to have slightly superior weight loss effect at a dose of 10 nmol/kg.
- GDF15 R&D Systems
- Xaa is Phe or Ala
- Xaa is Leu or Ala
- Xaa is absent or Lys
- Xaa is Ser or Pro
- gacaaaactc acacatgccc accgtgccca gcacctgaac tcctgggggg accgtcagtc 60
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Abstract
Description
아미노산 서열 | 서열번호 | ||
Signal Peptide | MHRPEAMLLL LTLALLGGPT WA | 17 | |
IgG1 Fc | Hinge | DKTHTCPPCP | 4 |
CH2-CH3 | APELLGGPSV FLFPPKPKDT LMISRTPEVT CVVVDVSHED PEVKFNWYVD GVEVHNAKTK PREEQYNSTY RVVSVLTVLH QDWLNGKEYK CKVSNKALPA PIEKTISKAK GQPREPQVYT LPPSRDELTK NQVSLTCLVK GFYPSDIAVE WESNGQPENN YKTTPPVLDS DGSFFLYSKL TVDKSRWQQG NVFSCSVMHE ALHNHYTQKS LSLSPG | 3 | |
Linker | GGGGSGGGGS GGGGSGGGGS | 18 | |
GDF(CRL) | CRLHTVRASL EDLGWADWVL SPREVQVTMC IGACPSQFRA ANMHAQIKTS LHRLKPDTVP APCCVPASYN PMVLIQKTDT GVSLQTYDDL LAKDCHCI | 2 |
아미노산 서열 | 서열번호 | ||
Signal Peptide | MHRPEAMLLL LTLALLGGPT WA | 17 | |
Mutated IgG4 Fc | Hinge | ESKYGPPCPP CP | 12 |
CH2-CH3 | APEAAGGPSV FLFPPKPKDT LMISRTPEVT CVVVDVSQED PEVQFNWYVD GVEVHNAKTK PREEQFNSTY RVVSVLTVLH QDWLNGKEYK CKVSNKGLPS SIEKTISKAK GQPREPQVYT LPPSQEEMTK NQVSLTCLVK GFYPSDIAVE WESNGQPENN YKTTPPVLDS DGSFFLYSRL TVDKSRWQEG NVFSCSVMHE ALHNHYTQKS LSLSLG | 7 | |
Linker | GGGGSGGGGS GGGGSGGGGS | 18 | |
GDF15 | ARNGDHCPLG PGRCCRLHTV RASLEDLGWA DWVLSPREVQ VTMCIGACPS QFRAANMHAQ IKTSLHRLKP DTVPAPCCVP ASYNPMVLIQ KTDTGVSLQT YDDLLAKDCH CI | 1 |
아미노산 서열 | 서열번호 | ||
Signal Peptide | MHRPEAMLLL LTLALLGGPT WA | 17 | |
Mutated IgG4 Fc | Hinge | ESKYGPPCPP CP | 12 |
CH2-CH3 | APEAAGGPSV FLFPPKPKDT LMISRTPEVT CVVVDVSQED PEVQFNWYVD GVEVHNAKTK PREEQFNSTY RVVSVLTVLH QDWLNGKEYK CKVSNKGLPS SIEKTISKAK GQPREPQVYT LPPSQEEMTK NQVSLTCLVK GFYPSDIAVE WESNGQPENN YKTTPPVLDS DGSFFLYSRL TVDKSRWQEG NVFSCSVMHE ALHNHYTQKS LSLSLG | 7 | |
Linker | GGGGSGGGGS GGGGSGGGGS | 18 | |
GDF15(CRL) | CRLHTVRASL EDLGWADWVL SPREVQVTMC IGACPSQFRA ANMHAQIKTS LHRLKPDTVP APCCVPASYN PMVLIQKTDT GVSLQTYDDL LAKDCHCI | 2 |
시료명 | 흡광계수 (0.1%, 1 mg/mL) |
IgG1-GDF (CRL) | 1.317 |
HIgG1-GDF (CRL) | 1.22 |
IgG4-GDF (CRL) | 1.334 |
IgG4-GDF | 1.287 |
High Fat Diet | 시험물질 | 투여경로 | 투여용량 (nmol/kg) | 투여볼륨 (mL/kg) | 동물수 |
X | Vehicle | 피하 | - | 5 | 5 |
O | Vehicle, qd | 피하 | - | 5 | 5 |
O | Vehicle, qw | 피하 | - | 5 | 5 |
O | GDF15 | 피하 | 2.86 | 5 | 3 |
O | IgG1-GDF (CRL) | 피하 | 1 | 5 | 5 |
O | IgG1-GDF (CRL) | 피하 | 10 | 5 | 5 |
O | HIgG1-GDF (CRL) | 피하 | 1 | 5 | 5 |
O | HIgG1-GDF (CRL) | 피하 | 10 | 5 | 5 |
관찰항목 | 순화기간(week) | 기간(day) | ||||||||
1 | 2 | 0 | 1 | 2 | 3 | 4 | 5 | 6 | 7 | |
고지방 사료 급이 | ● | ● | ● | ● | ● | ● | ● | ● | ● | ● |
경구 및 피하 투여 적응 | ● | |||||||||
투여 | ● | |||||||||
체중측정 | ● | ● | ● | ● | ● | ● | ● | ● | ● | |
사료섭취량 측정 | ● | ● | ● | ● | ● | ● | ● | ● |
High Fat Diet | 시험물질 | 투여경로 | 투여용량 (nmol/kg) | 투여볼륨 (mL/kg) | 동물수 |
O | Vehicle, qw | 피하 | - | 5 | 5 |
O | Semaglutide, qd | 피하 | 3 | 5 | 5 |
O | IgG4-GDF | 피하 | 1 | 5 | 5 |
O | IgG4-GDF | 피하 | 10 | 5 | 5 |
O | IgG4-GDF (CRL) | 피하 | 1 | 5 | 5 |
O | IgG4-GDF (CRL) | 피하 | 10 | 5 | 5 |
Group | Day | 0 | 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 |
Lean Vehicle Control (Daily inj.) | Mean | 100 | 102 | 101 | 101 | 101 | 101 | 101 | 101 | 101 | 102 |
S.E. | 0 | 1 | 1 | 1 | 1 | 1 | 1 | 1 | 1 | 1 | |
DIO Vehicle Control (Daily inj.) | Mean | 100 | 100 | 100 | 100 | 100 | 100 | 101 | 101 | 101 | 101 |
S.E. | 0 | 0 | 0 | 0 | 1 | 1 | 1 | 1 | 0 | 0 | |
DIO Vehicle Control (Single Inj.) | Mean | 100 | 100 | 99 | 100 | 100 | 100 | 101 | 101 | 102 | 102 |
S.E. | 0 | 0 | 0 | 0 | 1 | 1 | 1 | 1 | 1 | 1 | |
GDF15, 2.86 nmol/kg (Daily Inj.) | Mean | 100 | 99 | 98 | 97 | 96 | 95 | 95 | 95 | 96 | 97 |
S.E. | 0 | 0 | 0 | 1 | 1 | 1 | 1 | 2 | 2 | 2 | |
IgG1-GDF(CRL), 1 nmol/kg (Single Inj.) | Mean | 100 | 99 | 98 | 96 | 96 | 96 | 95 | 95 | 95 | 95 |
S.E. | 0 | 1 | 1 | 0 | 1 | 1 | 1 | 1 | 1 | 1 | |
IgG1-GDF(CRL), 10 nmol/kg (Single Inj.) | Mean | 100 | 98 | 97 | 95 | 94 | 93 | 91 | 90 | 90 | 90 |
S.E. | 0 | 0 | 1 | 1 | 1 | 1 | 1 | 1 | 1 | 1 | |
HIgG1-GDF(CRL), 1 nmol/kg (Single Inj.) | Mean | 100 | 100 | 98 | 98 | 97 | 96 | 97 | 96 | 96 | 97 |
S.E. | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 1 | |
HIgG1-GDF(CRL), 10 nmol/kg (Single Inj.) | Mean | 100 | 99 | 98 | 97 | 95 | 95 | 94 | 93 | 93 | 93 |
S.E. | 0 | 0 | 0 | 0 | 1 | 1 | 1 | 1 | 1 | 1 |
Group | Day | 0 | 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 |
DIO Vehicle Control (Single Inj.) | Mean | 100 | 100 | 100 | 99 | 100 | 100 | 100 | 100 | 101 | 101 |
S.E. | 0 | 0 | 1 | 0 | 0 | 0 | 0 | 1 | 0 | 1 | |
Semaglutide, 3 nmol/kg (Daily Inj.) | Mean | 100 | 94 | 91 | 90 | 87 | 87 | 85 | 86 | 83 | 85 |
S.E. | 0 | 0 | 1 | 2 | 2 | 3 | 3 | 3 | 3 | 3 | |
IgG4-GDF15, 1 nmol/kg (Single Inj.) | Mean | 100 | 99 | 97 | 96 | 95 | 93 | 93 | 92 | 92 | 91 |
S.E. | 0 | 0 | 1 | 1 | 1 | 1 | 1 | 1 | 1 | 1 | |
IgG4-GDF15, 10 nmol/kg (Single Inj.) | Mean | 100 | 98 | 97 | 96 | 94 | 93 | 93 | 92 | 91 | 90 |
S.E. | 0 | 0 | 1 | 1 | 1 | 1 | 1 | 1 | 2 | 2 | |
IgG4-GDF(CRL), 1 nmol/kg (Single Inj.) | Mean | 100 | 99 | 98 | 97 | 96 | 95 | 94 | 93 | 92 | 92 |
S.E. | 0 | 0 | 0 | 0 | 1 | 1 | 1 | 1 | 1 | 1 | |
IgG4-GDF(CRL), 10 nmol/kg (Single Inj.) | Mean | 100 | 98 | 97 | 95 | 94 | 92 | 91 | 90 | 88 | 87 |
S.E. | 0 | 0 | 0 | 1 | 1 | 1 | 1 | 1 | 1 | 1 | |
Group | Day | 10 | 11 | 12 | 13 | 14 | 15 | 16 | 17 | 18 | 19 |
DIO Vehicle Control (Single Inj.) | Mean | 102 | 102 | 103 | 104 | 104 | 104 | 105 | 105 | 105 | 105 |
S.E. | 1 | 1 | 0 | 1 | 1 | 1 | 1 | 1 | 1 | 1 | |
Semaglutide, 3 nmol/kg (Daily Inj.) | Mean | 84 | 82 | 81 | 82 | 81 | 79 | 79 | 78 | 77 | 77 |
S.E. | 3 | 3 | 3 | 3 | 3 | 2 | 2 | 2 | 2 | 1 | |
IgG4-GDF15, 1 nmol/kg (Single Inj.) | Mean | 91 | 91 | 93 | 94 | 94 | 94 | 95 | 96 | 96 | 96 |
S.E. | 1 | 1 | 1 | 1 | 1 | 1 | 1 | 2 | 1 | 2 | |
IgG4-GDF15, 10 nmol/kg (Single Inj.) | Mean | 90 | 90 | 91 | 92 | 92 | 92 | 92 | 93 | 94 | 94 |
S.E. | 2 | 1 | 2 | 2 | 2 | 2 | 2 | 2 | 2 | 2 | |
IgG4-GDF(CRL), 1 nmol/kg (Single Inj.) | Mean | 91 | 91 | 91 | 91 | 91 | 91 | 91 | 92 | 92 | 92 |
S.E. | 1 | 1 | 1 | 1 | 1 | 1 | 1 | 2 | 2 | 2 | |
IgG4-GDF(CRL), 10 nmol/kg (Single Inj.) | Mean | 85 | 85 | 84 | 83 | 83 | 82 | 83 | 84 | 84 | 85 |
S.E. | 1 | 1 | 1 | 1 | 2 | 2 | 2 | 2 | 3 | 2 |
Group | Day | 0 | 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 |
Lean Vehicle Control (Daily inj.) | Mean | 4.5 | 4.6 | 4.6 | 4.3 | 4.4 | 4.6 | 4.2 | 4.5 | 3.9 | 4.7 |
S.E. | 0.2 | 0.2 | 0.1 | 0.2 | 0.1 | 0.3 | 0.2 | 0.2 | 0.1 | 0.3 | |
DIO Vehicle Control (Daily inj.) | Mean | 2.8 | 2.8 | 3.1 | 3.0 | 3.1 | 2.9 | 3.0 | 2.9 | 2.9 | 3.3 |
S.E. | 0.3 | 0.2 | 0.2 | 0.3 | 0.3 | 0.2 | 0.3 | 0.2 | 0.1 | 0.2 | |
DIO Vehicle Control (Single Inj.) | Mean | 3.1 | 2.9 | 3.1 | 3.2 | 3.3 | 3.2 | 3.3 | 3.2 | 3.0 | 3.5 |
S.E. | 0.1 | 0.2 | 0.2 | 0.2 | 0.2 | 0.1 | 0.1 | 0.1 | 0.1 | 0.2 | |
GDF15, 2.86 nmol/kg (Daily Inj.) | Mean | 2.8 | 2.4 | 2.7 | 2.4 | 2.5 | 2.6 | 2.7 | 3.0 | 3.3 | 3.5 |
S.E. | 0.4 | 0.3 | 0.2 | 0.4 | 0.2 | 0.3 | 0.2 | 0.3 | 0.2 | 0.3 | |
IgG1-GDF(CRL), 1 nmol/kg (Single Inj.) | Mean | 3.0 | 2.1 | 2.2 | 2.2 | 2.6 | 2.6 | 2.6 | 2.8 | 2.7 | 3.0 |
S.E. | 0.2 | 0.2 | 0.1 | 0.1 | 0.2 | 0.1 | 0.0 | 0.1 | 0.1 | 0.3 | |
IgG1-GDF(CRL), 10 nmol/kg (Single Inj.) | Mean | 2.7 | 2.0 | 2.5 | 2.0 | 1.9 | 1.9 | 2.1 | 2.1 | 2.3 | 2.8 |
S.E. | 0.3 | 0.1 | 0.1 | 0.3 | 0.2 | 0.2 | 0.2 | 0.1 | 0.1 | 0.3 | |
HIgG1-GDF(CRL), 1 nmol/kg (Single Inj.) | Mean | 3.0 | 2.4 | 2.4 | 2.5 | 2.7 | 2.6 | 2.9 | 2.7 | 2.9 | 3.0 |
S.E. | 0.2 | 0.1 | 0.1 | 0.0 | 0.1 | 0.0 | 0.1 | 0.1 | 0.1 | 0.1 | |
HIgG1-GDF(CRL), 10 nmol/kg (Single Inj.) | Mean | 3.8 | 2.2 | 2.2 | 2.3 | 2.4 | 2.4 | 2.3 | 2.4 | 2.9 | 3.3 |
S.E. | 0.5 | 0.1 | 0.1 | 0.1 | 0.2 | 0.1 | 0.1 | 0.1 | 0.3 | 0.3 |
Group | Day | 0 | 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 |
DIO Vehicle Control (Single Inj.) | Mean | 3.3 | 2.7 | 3.0 | 3.0 | 3.1 | 3.5 | 3.0 | 3.5 | 3.3 | 3.4 |
S.E. | 0.1 | 0.1 | 0.2 | 0.1 | 0.1 | 0.2 | 0.2 | 0.1 | 0.1 | 0.2 | |
Semaglutide, 3 nmol/kg (Daily Inj.) | Mean | 3.7 | 2.1 | 3.3 | 2.8 | 2.3 | 2.9 | 2.7 | 3.2 | 2.4 | 3.7 |
S.E. | 0.3 | 0.7 | 1.4 | 0.6 | 0.4 | 0.9 | 0.6 | 0.3 | 0.4 | 0.1 | |
IgG4-GDF15, 1 nmol/kg (Single Inj.) | Mean | 3.0 | 1.9 | 1.9 | 2.1 | 2.0 | 2.0 | 2.1 | 2.4 | 2.4 | 2.5 |
S.E. | 0.1 | 0.1 | 0.1 | 0.2 | 0.2 | 0.1 | 0.1 | 0.2 | 0.2 | 0.2 | |
IgG4-GDF15, 10 nmol/kg (Single Inj.) | Mean | 3.0 | 1.7 | 2.0 | 2.0 | 2.1 | 2.2 | 2.4 | 2.4 | 2.2 | 2.7 |
S.E. | 0.1 | 0.1 | 0.3 | 0.2 | 0.2 | 0.3 | 0.2 | 0.2 | 0.2 | 0.2 | |
IgG4-GDF(CRL), 1 nmol/kg (Single Inj.) | Mean | 3.3 | 2.0 | 2.6 | 2.7 | 2.4 | 2.7 | 2.3 | 2.5 | 2.5 | 2.8 |
S.E. | 0.2 | 0.1 | 0.2 | 0.2 | 0.1 | 0.1 | 0.2 | 0.1 | 0.2 | 0.1 | |
IgG4-GDF(CRL), 10 nmol/kg (Single Inj.) | Mean | 3.1 | 1.5 | 2.3 | 2.4 | 2.4 | 2.3 | 2.3 | 2.0 | 2.3 | 2.4 |
S.E. | 0.2 | 0.3 | 0.2 | 0.2 | 0.2 | 0.2 | 0.2 | 0.1 | 0.2 | 0.3 | |
Group | Day | 10 | 11 | 12 | 13 | 14 | 15 | 16 | 17 | 18 | 19 |
DIO Vehicle Control (Single Inj.) | Mean | 3.0 | 3.4 | 3.2 | 3.5 | 3.1 | 3.3 | 3.1 | 3.2 | 3.3 | 3.3 |
S.E. | 0.2 | 0.1 | 0.2 | 0.1 | 0.1 | 0.2 | 0.1 | 0.1 | 0.1 | 0.1 | |
Semaglutide, 3 nmol/kg (Daily Inj.) | Mean | 2.7 | 2.5 | 2.4 | 3.0 | 2.5 | 2.1 | 2.5 | 2.0 | 2.2 | 3.7 |
S.E. | 0.4 | 0.4 | 0.3 | 0.4 | 0.3 | 0.3 | 0.2 | 0.2 | 0.2 | 0.3 | |
IgG4-GDF15, 1 nmol/kg (Single Inj.) | Mean | 2.7 | 3.1 | 3.2 | 3.7 | 3.0 | 3.1 | 3.1 | 3.1 | 3.1 | 3.0 |
S.E. | 0.1 | 0.0 | 0.0 | 0.1 | 0.1 | 0.3 | 0.2 | 0.3 | 0.1 | 0.1 | |
IgG4-GDF15, 10 nmol/kg (Single Inj.) | Mean | 3.1 | 3.8 | 3.3 | 3.7 | 3.4 | 3.6 | 3.1 | 3.5 | 3.3 | 3.0 |
S.E. | 0.3 | 0.7 | 0.5 | 0.5 | 0.4 | 0.6 | 0.2 | 0.1 | 0.1 | 0.1 | |
IgG4-GDF(CRL), 1 nmol/kg (Single Inj.) | Mean | 2.4 | 2.8 | 2.6 | 3.0 | 2.8 | 2.9 | 3.1 | 3.3 | 3.0 | 3.3 |
S.E. | 0.1 | 0.1 | 0.2 | 0.2 | 0.2 | 0.2 | 0.2 | 0.2 | 0.2 | 0.2 | |
IgG4-GDF(CRL), 10 nmol/kg (Single Inj.) | Mean | 2.1 | 2.5 | 2.3 | 2.5 | 2.6 | 2.6 | 3.0 | 3.4 | 3.3 | 3.1 |
S.E. | 0.1 | 0.2 | 0.3 | 0.2 | 0.4 | 0.2 | 0.3 | 0.3 | 0.3 | 0.2 |
PK Parameter | Group 1 | Group 2 | Group 3 |
GDF15 | IgG4-GDF | IgG4-GDF(CRL) | |
Cmax (ug/mL) | 0.443 | 10.6 | 9.73 |
Tmax (hr) | 1 | 96 | 48 |
AUClast (ug·hr/mL) | 4.86 | 2144 | 2004 |
AUCinf (ug·hr/mL) | 4.88 | 2854 | 2593 |
t1/2 (hr) | 19.0 | 101 | 114 |
AUCextp (%) | 0.463 | 14.6 | 18.2 |
Claims (21)
- GDF15 (Growth/differentiation factor 15) 또는 이의 기능적 변이체, 및면역글로불린의 Fc 영역을 포함하고,상기 면역글로불린의 Fc 영역은 단일 사슬의 IgG1 Fc 영역 또는 IgG4 Fc 영역이고, 상기 GDF15 또는 이의 기능적 변이체의 N-말단에 유연성(flexible) 펩타이드 링커를 통하여 연결되며,상기 GDF15의 기능적 변이체는 서열번호 1의 아미노산 서열 중 1번째부터 14번째까지의 14개 아미노산 중 하나 이상이 결실된 결실 변이체이고,상기 유연성 펩타이드 링커는 (GGGGS)n (n은 1, 2, 3, 4, 또는 5)로 표현되는 것인,융합 폴리펩타이드.
- 제1항에 있어서, 상기 면역글로불린의 Fc 영역은 인간 IgG4 Fc 영역인, 융합 폴리펩타이드.
- 제2항에 있어서, 상기 인간 IgG4 Fc 영역은(1) 서열번호 5의 아미노산 서열을 포함하거나,(2) 서열번호 5의 아미노산 서열의 N 말단에 서열번호 10의 아미노산 서열을 추가로 포함하는 것인,융합 폴리펩타이드.
- 제3항에 있어서, 상기 인간 IgG4 Fc 영역은(1) 서열번호 6 내지 9 중에서 선택된 아미노산 서열을 포함하거나,(2) 서열번호 6 내지 9 중에서 선택된 아미노산 서열의 N 말단에 서열번호 11 또는 서열번호 12의 아미노산 서열을 추가로 포함하는 것인,융합 폴리펩타이드.
- 제1항에 있어서, 상기 GDF15의 기능적 변이체는 서열번호 2의 아미노산 서열을 포함하는 것인, 융합 폴리펩타이드.
- 제1항 내지 제5항 중 어느 한 항에 있어서, 상기 융합 폴리펩타이드 내의 면역글로불린의 Fc 영역과 연결된 GDF15 또는 이의 기능적 변이체는, 면역글로불린의 Fc 영역과 결합되지 않은 GDF15 또는 이의 기능적 변이체와 비교하여, 체내 반감기가 1.5배 이상 증가한 것인, 융합 폴리펩타이드.
- 제1항 내지 제5항 중 어느 한 항의 융합 폴리펩타이드를 2개 포함하는, 융합 폴리펩타이드 이량체.
- 제1항 내지 제5항 중 어느 한 항의 융합 폴리펩타이드를 암호화하는 핵산 분자.
- 제8항의 핵산 분자를 포함하는 재조합 벡터.
- 제9항의 재조합 벡터를 포함하는 재조합 세포.
- 제10항의 재조합 세포를 배양하는 단계를 포함하는, GDF15 또는 이의 기능적 변이체 및 면역글로불린의 Fc 영역을 포함하는, 제1항의 융합 폴리펩타이드의 제조 방법.
- 단일 사슬의 IgG1 Fc 영역 또는 IgG4 Fc 영역을 GDF15 또는 이의 기능적 변이체의 N 말단에 유연성 펩타이드 링커를 통하여 연결시키는 단계를 포함하고,상기 GDF15의 기능적 변이체는 서열번호 1의 아미노산 서열 중 1번째부터 14번째까지의 14개 아미노산 중 하나 이상이 결실된 결실 변이체이고,상기 유연성 펩타이드 링커는 (GGGGS)n [(서열번호 13)n; n은 1, 2, 3, 4, 또는 5]로 표현되는 것인,GDF15 또는 이의 기능적 변이체의 체내 안정성 증진 방법.
- 제12항에 있어서, 상기 면역글로불린의 Fc 영역은 인간 IgG4 Fc 영역인, GDF15 또는 이의 기능적 변이체의 체내 안정성 증진 방법.
- 제13항에 있어서, 상기 인간 IgG4 Fc 영역은(1) 서열번호 5의 아미노산 서열을 포함하거나,(2) 서열번호 5의 아미노산 서열의 N 말단에 서열번호 10의 아미노산 서열을 추가로 포함하는 것인,GDF15 또는 이의 기능적 변이체의 체내 안정성 증진 방법.
- 제14항에 있어서, 상기 인간 IgG4 Fc 영역은(1) 서열번호 6 내지 9 중에서 선택된 아미노산 서열을 포함하거나,(2) 서열번호 6 내지 9 중에서 선택된 아미노산 서열의 N 말단에 서열번호 11 또는 서열번호 12의 아미노산 서열을 추가로 포함하는 것인,GDF15 또는 이의 기능적 변이체의 체내 안정성 증진 방법.
- 제12항에 있어서, 상기 GDF15의 기능적 변이체는 서열번호 2의 아미노산 서열을 포함하는 것인, GDF15 또는 이의 기능적 변이체의 체내 안정성 증진 방법.
- 제12항 내지 제16항 중 어느 한 항에 있어서, 상기 면역글로불린의 Fc 영역과 연결된 GDF15 또는 이의 기능적 변이체는, 면역글로불린의 Fc 영역과 결합되지 않은 GDF15 또는 이의 기능적 변이체와 비교하여, 체내 반감기가 1.5배 이상 증가한 것인, GDF15 또는 이의 기능적 변이체의 체내 안정성 증진 방법.
- 제1항 내지 제5항 중 어느 한 항의 융합 폴리펩타이드, 상기 융합 폴리펩타이드를 2개 포함하는 융합 폴리펩타이드 이량체, 상기 융합 폴리펩타이드를 암호화하는 핵산 분자, 상기 핵산 분자를 포함하는 재조합 벡터, 및 상기 재조합 벡터를 포함하는 재조합 세포로 이루어진 군에서 선택된 하나 이상을 포함하는, 체중 감소용 조성물.
- 제1항 내지 제5항 중 어느 한 항의 융합 폴리펩타이드, 상기 융합 폴리펩타이드를 2개 포함하는 융합 폴리펩타이드 이량체, 상기 융합 폴리펩타이드를 암호화하는 핵산 분자, 상기 핵산 분자를 포함하는 재조합 벡터, 및 상기 재조합 벡터를 포함하는 재조합 세포로 이루어진 군에서 선택된 하나 이상을 포함하는, 식이 조절용 조성물.
- 제1항 내지 제5항 중 어느 한 항의 융합 폴리펩타이드, 상기 융합 폴리펩타이드를 2개 포함함하는 융합 폴리펩타이드 이량체, 상기 융합 폴리펩타이드를 암호화하는 핵산 분자, 상기 핵산 분자를 포함하는 재조합 벡터, 및 상기 재조합 벡터를 포함하는 재조합 세포로 이루어진 군에서 선택된 하나 이상을 포함하는, 대사성 질병의 예방 또는 치료용 약학 조성물.
- 제20항에 있어서, 상기 대사성 질병은 비만, 당뇨, 또는 비알코올성 지방간 질환인, 약학 조성물.
Priority Applications (15)
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AU2020260931A AU2020260931B2 (en) | 2019-04-23 | 2020-04-22 | Fusion polypeptide comprising Fc region of immunoglobulin and GDF15 |
MX2021012964A MX2021012964A (es) | 2019-04-23 | 2020-04-22 | Polipeptido de fusion que comprende region fc de inmunoglobulina y gdf15. |
JOP/2021/0282A JOP20210282A1 (ar) | 2019-04-23 | 2020-04-22 | بولي ببتيد اندماج يتضمن منطقة fc لجلوبيولين مناعي وgdf15 |
PE2021001749A PE20220500A1 (es) | 2019-04-23 | 2020-04-22 | POLIPEPTIDO DE FUSION QUE COMPRENDE LA REGION Fc DE INMUNOGLOBULINA Y GDF15 |
EP20793990.1A EP3978518A4 (en) | 2019-04-23 | 2020-04-22 | FUSION POLYPEPTIDE COMPRISING FC REGION OF IMMUNOGLOBULIN AND GDF15 |
JP2021562863A JP2022530216A (ja) | 2019-04-23 | 2020-04-22 | 免疫グロブリンのFc領域およびGDF15を含む融合ポリペプチド |
CA3136969A CA3136969A1 (en) | 2019-04-23 | 2020-04-22 | Fusion polypeptide comprising fc region of immunoglobulin and gdf15 |
US17/605,798 US20230053119A1 (en) | 2019-04-23 | 2020-04-22 | FUSION POLYPEPTIDE COMPRISING Fc REGION OF IMMUNOGLOBULIN AND GDF15 |
BR112021021271A BR112021021271A2 (pt) | 2019-04-23 | 2020-04-22 | Polipeptídeo de fusão compreendendo região fc de imunoglobulina e gdf15 |
CN202080030773.8A CN113767112A (zh) | 2019-04-23 | 2020-04-22 | 包含免疫球蛋白Fc区和GDF15的融合多肽 |
SG11202111570TA SG11202111570TA (en) | 2019-04-23 | 2020-04-22 | Fusion polypeptide comprising fc region of immunoglobulin and gdf15 |
IL287343A IL287343A (en) | 2019-04-23 | 2021-10-18 | A fused polypeptide containing the fc region of immunoglobulin and gdf15 |
CONC2021/0014413A CO2021014413A2 (es) | 2019-04-23 | 2021-10-27 | Polipéptido de fusión que comprende región fc de inmunoglobulina y gdf15 |
ZA2021/08437A ZA202108437B (en) | 2019-04-23 | 2021-10-29 | Fusion polypeptide comprising fc region of immunoglobulin and gdf15 |
JP2023101082A JP2023115118A (ja) | 2019-04-23 | 2023-06-20 | 免疫グロブリンのFc領域およびGDF15を含む融合ポリペプチド |
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KR10-2019-0047558 | 2019-04-23 | ||
KR20190047558 | 2019-04-23 |
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PCT/KR2020/005324 WO2020218827A1 (ko) | 2019-04-23 | 2020-04-22 | 면역글로불린의 Fc 영역 및 GDF15를 포함하는 융합 폴리펩타이드 |
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US (1) | US20230053119A1 (ko) |
EP (1) | EP3978518A4 (ko) |
JP (2) | JP2022530216A (ko) |
KR (2) | KR20200124176A (ko) |
CN (1) | CN113767112A (ko) |
AU (1) | AU2020260931B2 (ko) |
BR (1) | BR112021021271A2 (ko) |
CA (1) | CA3136969A1 (ko) |
CL (1) | CL2021002757A1 (ko) |
CO (1) | CO2021014413A2 (ko) |
IL (1) | IL287343A (ko) |
JO (1) | JOP20210282A1 (ko) |
MX (1) | MX2021012964A (ko) |
PE (1) | PE20220500A1 (ko) |
SG (1) | SG11202111570TA (ko) |
TW (2) | TW202100563A (ko) |
WO (1) | WO2020218827A1 (ko) |
ZA (1) | ZA202108437B (ko) |
Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2023025123A1 (zh) * | 2021-08-24 | 2023-03-02 | 广东东阳光药业有限公司 | Gdf15融合蛋白及其用途 |
WO2023154953A1 (en) | 2022-02-14 | 2023-08-17 | Ngm Biopharmaceuticals, Inc. | Gdf15 polypeptides for treating and preventing autoimmune diseases |
Families Citing this family (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2022245183A1 (en) * | 2021-05-21 | 2022-11-24 | Yuhan Corporation | Composition for preventing or treating non-alcoholic fatty liver disease or non-alcoholic steatohepatitis comprising growth differentiation factor-15 variant |
Citations (8)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
KR101038126B1 (ko) | 2010-11-30 | 2011-05-31 | 주식회사 엘지생명과학 | 새로운 융합 프로모터 및 이를 포함하는 재조합 벡터 |
US20140378665A1 (en) * | 2012-01-26 | 2014-12-25 | Amgen Inc. | Growth differentiation factor 15 (gdf-15) polypeptides |
KR20150125402A (ko) | 2014-04-30 | 2015-11-09 | 주식회사 엘지생명과학 | 고효율 분비능을 가지는 단백질 분비 인자 및 이를 포함하는 발현 벡터 |
US20160168213A1 (en) * | 2013-07-31 | 2016-06-16 | Amgen Inc. | Growth differentiation factor 15 (gdf-15) constructs |
KR20170020878A (ko) * | 2014-06-23 | 2017-02-24 | 노파르티스 아게 | 지방산 및 생체분자에 대한 접합에서의 이의 용도 |
EP3144320A1 (en) * | 2011-04-13 | 2017-03-22 | Bristol-Myers Squibb Company | Fc fusion proteins comprising novel linkers or arrangements |
KR101868139B1 (ko) | 2010-11-30 | 2018-06-15 | 주식회사 엘지화학 | 새로운 융합 프로모터 및 이를 포함하는 재조합 벡터 |
KR20190003746A (ko) * | 2016-05-10 | 2019-01-09 | 얀센 바이오테크 인코포레이티드 | Gdf15 융합 단백질 및 이의 용도 |
Family Cites Families (10)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
ATE240395T1 (de) * | 1994-03-29 | 2003-05-15 | Celltech Therapeutics Ltd | Antikörper gegen e-selektin |
WO2002079415A2 (en) * | 2001-03-30 | 2002-10-10 | Lexigen Pharmaceuticals Corp. | Reducing the immunogenicity of fusion proteins |
US7737260B2 (en) | 2003-11-13 | 2010-06-15 | Hanmi Pharm. Co., Ltd | Protein complex using an immunoglobulin fragment and method for the preparation thereof |
US9550819B2 (en) * | 2012-03-27 | 2017-01-24 | Ngm Biopharmaceuticals, Inc. | Compositions and methods of use for treating metabolic disorders |
JP6272907B2 (ja) * | 2013-01-30 | 2018-01-31 | エヌジーエム バイオファーマシューティカルズ インコーポレイテッド | 代謝障害の処置における組成物及び使用方法 |
US9920118B2 (en) * | 2014-10-31 | 2018-03-20 | Ngm Biopharmaceuticals, Inc. | Compositions and methods of use for treating metabolic disorders |
HUE061108T2 (hu) * | 2015-05-18 | 2023-05-28 | Pieris Pharmaceuticals Gmbh | Rákellenes fúziós polipeptid |
KR102668200B1 (ko) * | 2015-10-28 | 2024-05-23 | 주식회사유한양행 | 지속형 fgf21 융합 단백질 및 이를 포함하는 약학적 조성물 |
CN108367053A (zh) * | 2015-12-22 | 2018-08-03 | 诺华股份有限公司 | 使用生长分化因子15(gdf-15)治疗或改善代谢性疾病的方法 |
MX2020010659A (es) * | 2018-04-09 | 2020-10-28 | Amgen Inc | Proteinas de fusion del factor de diferenciacion de crecimiento 15. |
-
2020
- 2020-04-22 MX MX2021012964A patent/MX2021012964A/es unknown
- 2020-04-22 CN CN202080030773.8A patent/CN113767112A/zh active Pending
- 2020-04-22 JO JOP/2021/0282A patent/JOP20210282A1/ar unknown
- 2020-04-22 CA CA3136969A patent/CA3136969A1/en active Pending
- 2020-04-22 EP EP20793990.1A patent/EP3978518A4/en active Pending
- 2020-04-22 PE PE2021001749A patent/PE20220500A1/es unknown
- 2020-04-22 WO PCT/KR2020/005324 patent/WO2020218827A1/ko active Application Filing
- 2020-04-22 BR BR112021021271A patent/BR112021021271A2/pt unknown
- 2020-04-22 TW TW109113479A patent/TW202100563A/zh unknown
- 2020-04-22 AU AU2020260931A patent/AU2020260931B2/en active Active
- 2020-04-22 SG SG11202111570TA patent/SG11202111570TA/en unknown
- 2020-04-22 TW TW112122841A patent/TW202337914A/zh unknown
- 2020-04-22 KR KR1020200048925A patent/KR20200124176A/ko active Application Filing
- 2020-04-22 JP JP2021562863A patent/JP2022530216A/ja active Pending
- 2020-04-22 US US17/605,798 patent/US20230053119A1/en active Pending
-
2021
- 2021-06-24 KR KR1020210082625A patent/KR102609627B1/ko active IP Right Grant
- 2021-10-18 IL IL287343A patent/IL287343A/en unknown
- 2021-10-20 CL CL2021002757A patent/CL2021002757A1/es unknown
- 2021-10-27 CO CONC2021/0014413A patent/CO2021014413A2/es unknown
- 2021-10-29 ZA ZA2021/08437A patent/ZA202108437B/en unknown
-
2023
- 2023-06-20 JP JP2023101082A patent/JP2023115118A/ja active Pending
Patent Citations (8)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
KR101038126B1 (ko) | 2010-11-30 | 2011-05-31 | 주식회사 엘지생명과학 | 새로운 융합 프로모터 및 이를 포함하는 재조합 벡터 |
KR101868139B1 (ko) | 2010-11-30 | 2018-06-15 | 주식회사 엘지화학 | 새로운 융합 프로모터 및 이를 포함하는 재조합 벡터 |
EP3144320A1 (en) * | 2011-04-13 | 2017-03-22 | Bristol-Myers Squibb Company | Fc fusion proteins comprising novel linkers or arrangements |
US20140378665A1 (en) * | 2012-01-26 | 2014-12-25 | Amgen Inc. | Growth differentiation factor 15 (gdf-15) polypeptides |
US20160168213A1 (en) * | 2013-07-31 | 2016-06-16 | Amgen Inc. | Growth differentiation factor 15 (gdf-15) constructs |
KR20150125402A (ko) | 2014-04-30 | 2015-11-09 | 주식회사 엘지생명과학 | 고효율 분비능을 가지는 단백질 분비 인자 및 이를 포함하는 발현 벡터 |
KR20170020878A (ko) * | 2014-06-23 | 2017-02-24 | 노파르티스 아게 | 지방산 및 생체분자에 대한 접합에서의 이의 용도 |
KR20190003746A (ko) * | 2016-05-10 | 2019-01-09 | 얀센 바이오테크 인코포레이티드 | Gdf15 융합 단백질 및 이의 용도 |
Non-Patent Citations (1)
Title |
---|
KABAT, E.A. ET AL.: "Sequences of Proteins of Immunological Interest", 1991, U.S. DEPT. OF HEALTH AND HUMAN SERVICES |
Cited By (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2023025123A1 (zh) * | 2021-08-24 | 2023-03-02 | 广东东阳光药业有限公司 | Gdf15融合蛋白及其用途 |
WO2023025129A1 (zh) * | 2021-08-24 | 2023-03-02 | 广东东阳光药业有限公司 | Gdf15融合蛋白及其用途 |
WO2023154953A1 (en) | 2022-02-14 | 2023-08-17 | Ngm Biopharmaceuticals, Inc. | Gdf15 polypeptides for treating and preventing autoimmune diseases |
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JP2023115118A (ja) | 2023-08-18 |
KR102609627B1 (ko) | 2023-12-04 |
CA3136969A1 (en) | 2020-10-29 |
KR20210080339A (ko) | 2021-06-30 |
ZA202108437B (en) | 2023-07-26 |
US20230053119A1 (en) | 2023-02-16 |
AU2020260931B2 (en) | 2023-08-24 |
CN113767112A (zh) | 2021-12-07 |
TW202337914A (zh) | 2023-10-01 |
BR112021021271A2 (pt) | 2021-12-21 |
SG11202111570TA (en) | 2021-11-29 |
AU2020260931A1 (en) | 2021-12-09 |
JOP20210282A1 (ar) | 2023-01-30 |
CL2021002757A1 (es) | 2022-06-10 |
EP3978518A4 (en) | 2022-11-02 |
JP2022530216A (ja) | 2022-06-28 |
EP3978518A1 (en) | 2022-04-06 |
IL287343A (en) | 2021-12-01 |
KR20200124176A (ko) | 2020-11-02 |
MX2021012964A (es) | 2021-12-10 |
PE20220500A1 (es) | 2022-04-07 |
TW202100563A (zh) | 2021-01-01 |
CO2021014413A2 (es) | 2021-11-19 |
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