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CN-121986112-A - Bivalent and multivalent albumin binders

CN121986112ACN 121986112 ACN121986112 ACN 121986112ACN-121986112-A

Abstract

The present technology relates to bivalent and multivalent albumin binders. In particular, the present technology relates to novel and improved human serum albumin binders, in particular to polypeptides comprising (i) at least one Immunoglobulin Single Variable Domain (ISVD) that specifically binds to serum albumin or at least one human serum albumin and (ii) at least one further moiety that specifically binds to serum albumin or at least one human serum albumin.

Inventors

  • T. Van Bogart
  • J. Verkhurst

Assignees

  • 阿布林克斯有限公司
  • 赛诺菲

Dates

Publication Date
20260505
Application Date
20240920
Priority Date
20230922

Claims (15)

  1. 1. A polypeptide comprising: At least one Immunoglobulin Single Variable Domain (ISVD) that specifically binds to serum albumin, and at least one additional moiety that specifically binds to serum albumin or at least one human serum albumin, Wherein said (i) at least one ISVD consists essentially of 4 framework regions (FR 1 to FR4, respectively) and three complementarity determining regions (CDR 1 to CDR3, respectively), wherein: CDR1 comprises the amino acid sequence of SEQ ID NO. 1or has 3, 2 or 1 amino acid differences with SEQ ID NO.1, or CDR1 comprises the amino acid sequence of SEQ ID NO. 22 or has 3, 2 or 1 amino acid differences with SEQ ID NO. 22; CDR2 comprises the amino acid sequence of SEQ ID NO. 2 or has 3, 2 or 1 amino acid differences from SEQ ID NO. 2, and CDR3 comprises the amino acid sequence of SEQ ID NO. 3 or has 3, 2 or 1 amino acid differences with SEQ ID NO. 3, And/or CDR1 comprises the amino acid sequence of SEQ ID NO. 36 or has 3, 2 or 1 amino acid differences from SEQ ID NO. 36; CDR2 comprises the amino acid sequence of SEQ ID NO. 37 or has 3,2 or 1 amino acid differences from SEQ ID NO. 37 and CDR3 comprises the amino acid sequence of SEQ ID NO. 38 or has 3, 2 or 1 amino acid differences from SEQ ID NO. 38, And/or CDR1 comprises the amino acid sequence of SEQ ID NO. 55 or has 3, 2 or 1 amino acid differences from SEQ ID NO. 55; CDR2 comprises the amino acid sequence of SEQ ID NO: 56 or has 3,2 or 1 amino acid differences from SEQ ID NO: 56 and CDR3 comprises the amino acid sequence of SEQ ID NO: 57 or has 3, 2 or 1 amino acid differences from SEQ ID NO: 57 Wherein the CDR sequences are determined according to AbM numbering.
  2. 2. The polypeptide of claim 1, wherein the (i) at least one ISVD that specifically binds to serum albumin comprises: CDR1 comprising the amino acid sequence of SEQ ID NO. 1 or 22, CDR2 comprising the amino acid sequence of SEQ ID NO. 2, and CDR3 comprising the amino acid sequence of SEQ ID NO. 3, or CDR1 comprising the amino acid sequence of SEQ ID NO. 36, CDR2 comprising the amino acid sequence of SEQ ID NO. 37, and CDR3 comprising the amino acid sequence of SEQ ID NO. 38, or CDR1 comprising the amino acid sequence of SEQ ID NO. 55, CDR2 comprising the amino acid sequence of SEQ ID NO. 56, and CDR3 comprising the amino acid sequence of SEQ ID NO. 57; wherein the CDR sequences are determined according to AbM numbering.
  3. 3. The polypeptide of any one of claims 1-2, wherein the (i) specifically binds to at least one ISVD of serum albumin A degree of sequence identity of at least 85%, preferably at least 90%, more preferably at least 95% with any of the sequences as defined in SEQ ID NO 4 to 21, 54 or 91 to 93, and even more preferably a degree of sequence identity of 100% with any of the sequences as defined in SEQ ID NO 4 to 21, 54 or 91 to 93, wherein any C-terminal extension and these CDRs that may be present are not considered for determining the degree of sequence identity, and/or Has NO more than 7, preferably NO more than 5 amino acid differences, for example only 3, 2 or 1 amino acid differences, from any of the sequences as defined in SEQ ID NOS.4 to 21, 54 or 91 to 93.
  4. 4. The polypeptide of any one of claims 1-3, wherein the (i) at least one ISVD comprised therein is derived from V H , humanized V H , human V H 、V HH , humanized V HH , or camelized V H (derived from heavy chain ISVD).
  5. 5. The polypeptide of any one of claims 1 to 4, wherein the polypeptide comprises: at least one Immunoglobulin Single Variable Domain (ISVD) that specifically binds to serum albumin as defined in any of claims 1 to 5, and At least one human serum albumin.
  6. 6. The polypeptide of any one of claims 1 to 5, wherein the polypeptide comprises: at least one Immunoglobulin Single Variable Domain (ISVD) that specifically binds to serum albumin as defined in any of claims 1 to 5, and Specifically binds to at least one further portion of serum albumin, Wherein the (ii) specifically binds to at least one further moiety of serum albumin is selected from the group consisting of an antibody or fragment thereof, a single chain variable fragment (scFv), preferably an Immunoglobulin Single Variable Domain (ISVD), DARP-in, an affibody, an acilin and an Albumin Binding Domain (ABD).
  7. 7. The polypeptide of any one of claims 1 to 6, wherein the at least one additional moiety that specifically binds to serum albumin is ISVD comprising three complementarity determining regions (CDR 1 to CDR3, respectively), wherein: CDR1 comprises the amino acid sequence of SEQ ID NO. 1or has 3, 2 or 1 amino acid differences with SEQ ID NO.1, or CDR1 comprises the amino acid sequence of SEQ ID NO. 22 or has 3, 2 or 1 amino acid differences with SEQ ID NO. 22; CDR2 comprises the amino acid sequence of SEQ ID NO. 2 or has 3, 2 or 1 amino acid differences from SEQ ID NO. 2, and CDR3 comprises the amino acid sequence of SEQ ID NO. 3 or has 3, 2 or 1 amino acid differences with SEQ ID NO. 3, And/or CDR1 comprises the amino acid sequence of SEQ ID NO. 36 or has 3, 2 or 1 amino acid differences from SEQ ID NO. 36; CDR2 comprises the amino acid sequence of SEQ ID NO. 37 or has 3,2 or 1 amino acid differences from SEQ ID NO. 37 and CDR3 comprises the amino acid sequence of SEQ ID NO. 38 or has 3, 2 or 1 amino acid differences from SEQ ID NO. 38, And/or CDR1 comprises the amino acid sequence of SEQ ID NO. 55 or has 3, 2 or 1 amino acid differences from SEQ ID NO. 55; CDR2 comprises the amino acid sequence of SEQ ID NO: 56 or has 3,2 or 1 amino acid differences from SEQ ID NO: 56 and CDR3 comprises the amino acid sequence of SEQ ID NO. 57 or has 3, 2 or 1 amino acid differences from SEQ ID NO. 57, Wherein the CDR sequences are determined according to AbM numbering.
  8. 8. The polypeptide of claim 7, wherein the at least one additional moiety that specifically binds to serum albumin is ISVD A degree of sequence identity of at least 85%, preferably at least 90%, more preferably at least 95% to any of the sequences as defined in SEQ ID nos. 4 to 21, 54 or 91 to 93, wherein any C-terminal extension and the CDRs which may be present are not considered for determining the degree of sequence identity; And/or Having NO more than 7, preferably NO more than 5 amino acid differences, for example only 3, 2 or 1 amino acid differences, from any of the sequences as defined in SEQ ID NOS.4 to 21, 54 or 91 to 93, Preferably wherein the at least one further moiety that (ii) specifically binds to serum albumin is an ISVD having a sequence selected from SEQ ID NOs 4 to 21, 54 or 91 to 93.
  9. 9. A fusion protein comprising the polypeptide of any one of claims 1 to 8, wherein the fusion protein further comprises one or more (e.g., one or two) other amino acid sequences, (binding) domains, binding units or other moieties or chemical entities.
  10. 10. The fusion protein of claim 9, wherein the fusion protein comprises the polypeptide of any one of claims 1 to 8 and at least one therapeutic and/or targeting moiety, optionally wherein the therapeutic and/or targeting moiety comprises at least one ISVD that specifically binds to a therapeutic target, or wherein the fusion protein further comprises an additional ISVD that specifically binds to (human) serum albumin.
  11. 11. A composition comprising the polypeptide of any one of claims 1 to 8 and/or the fusion protein of any one of claims 9 to 10, optionally wherein the composition is a pharmaceutical composition.
  12. 12. A nucleic acid or nucleic acid sequence encoding a polypeptide according to any one of claims 1 to 8 and/or a fusion protein according to any one of claims 9 to 10.
  13. 13. A vector comprising the nucleic acid or nucleic acid sequence of claim 12.
  14. 14. A non-human host or host cell comprising the vector comprising a nucleic acid sequence of claim 13 or expressing the nucleic acid sequence of claim 12.
  15. 15. The polypeptide according to any one of claims 1 to 8, the fusion protein according to any one of claims 9 to 10 and/or the composition according to claim 11 for use as a medicament.

Description

Bivalent and multivalent albumin binders Technical Field The present technology relates to bivalent and multivalent albumin binders. In particular, the present technology relates to novel and improved human serum albumin binders, in particular to polypeptides comprising (i) at least one Immunoglobulin Single Variable Domain (ISVD) that specifically binds to serum albumin or at least one human serum albumin and (ii) at least one further moiety that specifically binds to serum albumin or at least one human serum albumin. The technology further relates to polypeptides comprising one albumin binding agent and one Human Serum Albumin (HSA) molecule, and to polypeptides comprising two HSA molecules. The present technology further relates to fusion proteins comprising polypeptides, to nucleic acids encoding such molecules or parts of such molecules, to host cells comprising such nucleic acids and/or expressing or being capable of expressing such polypeptides and/or fusion proteins, to compositions, and in particular to pharmaceutical compositions comprising such polypeptides and/or fusion proteins, nucleic acids and/or host cells. Furthermore, the present technology relates to methods for producing such polypeptides and fusion proteins, and to the use of such polypeptides and/or fusion proteins in different applications, including but not limited to extending the in vivo half-life of therapeutic compounds and/or preventing and/or treating diseases and/or disorders. Background Peptides and proteins are two classes of molecules that have attractive possibilities for therapeutic applications. However, the bottleneck in the development of peptides and proteins into clinically and commercially relevant drugs is their short half-life in vivo, typically only a few minutes to a few hours. The half-life of peptides and proteins in human serum is determined by several factors including size, charge, proteolytic sensitivity, their biological properties, turnover rate of the proteins to which they bind, and the like. There is a relationship between the hydrodynamic radius of a peptide or protein and its serum half-life. Generally, those peptides and proteins having a molecular weight of less than about 70 kDa are eliminated primarily by renal filtration and generally have a very short serum half-life. Larger proteins may last for several days in the circulation. Human Serum Albumin (HSA) and IgG are the two most abundant soluble proteins present in the blood circulation, and unlike most proteins in the circulation, they have a common property of extending serum half-life in humans by about 19 to 21 days. HSA is the most abundant plasma protein in blood and is a carrier protein involved in many processes to maintain homeostasis (i.e., maintain osmotic pressure). Albumin is widely used as a drug delivery vehicle （Mishra, V.;Heath, R.J. Structural and Biochemical Features of Human Serum Albumin Essential for Eukaryotic Cell Culture. [ for eukaryotic cell culture for its structural and biochemical characteristics of human serum albumin int.j.mol.sci. [ journal of international molecular sciences ] 2021, 22, 8411) due to its high serum concentration, long half-life, non-toxicity and low immunogenicity, and its ability to be taken in benign and tissues and bind to a variety of drugs. This prolongation of serum albumin half-life is mainly due to its protection from intracellular lysosomal degradation by binding to neonatal Fc receptor (FcRn). FcRn is a heterodimer consisting of an N-glycosylated transmembrane MHC class I-like heavy chain non-covalently associated with a soluble b 2-microglobulin. Both IgG and albumin are ligands that bind to different epitopes of FcRn. In general, fcRn recycling mechanisms are strictly pH dependent and favor binding to FcRn at low pH after acidification of the endosomal compartment (e.g., acidic endosomal pH, which is typically below 6.5). Albumin escapes degradation in lysosomes when it binds to FcRn. Upon returning to the cell surface, binding is reduced at extracellular physiological pH (typically around pH 7.4), resulting in albumin release into the blood stream (see, e.g., ward ES, ober RJ., targeting FcRn to Generate Antibody-Based therapeutics [ targeting FcRn to produce antibody-Based therapeutics ] Trends Pharmacol Sci. [ pharmacological Trends ], 2018; 39 (10): 892-904 and Andersen et al, extending Serum Half-life of Albumin by Engineering Neonatal Fc Receptor (FcRn) Binding [ extending serum half-life of albumin by engineered neonatal Fc receptor (FcRn) Binding ], JBC [ journal of biochemistry ], 2014, 289, 19: 13492-13502). In particular, albumin is increasingly being used to improve the pharmacokinetics of short-lived small molecule drugs that are capable of binding to albumin and biologically active therapeutic peptides and proteins by fusion of these molecules to the N-or C-terminal genes of albumin (Nilsen, j, trabjerg, e., grevys, a. Et al AN INTACT C-TERMINAL END of albumin is requ