US-20260125443-A1 - FUSION PROTEIN COMPRISING FOLLICLE STIMULATING HORMONE AND ANTIGEN-BINDING FRAGMENT TO SERUM ALBUMIN AND COMPOSITIONS AND USES THEREOF

US20260125443A1US 20260125443 A1US20260125443 A1US 20260125443A1US-20260125443-A1

Abstract

The present application relates to a fusion protein including a follicle stimulating hormone and an antigen-binding fragment to serum albumin and a use thereof. Accordingly, a recombinant fusion protein, which includes an antigen-binding fragment that binds to serum albumin and a human follicle stimulating hormone linked to the antigen-binding fragment, and a long-acting human follicle stimulating hormone formulation including the recombinant fusion protein as an active ingredient are provided.

Inventors

Sang Hoon Cha

Assignees

APRILBIO CO., LTD.

Dates

Publication Date: 20260507
Application Date: 20231004
Priority Date: 20221004

Claims (20)

1 . A recombinant fusion protein comprising (a) a human follicle stimulating hormone (hFSH) comprising an hFSH alpha subunit and an hFSH beta subunit and (b) an antigen binding fragment (Fab) that binds to serum albumin.
2 . The recombinant fusion protein of claim 1 , wherein the hFSH alpha subunit and the hFSH beta subunit are linked to terminal regions of the Fab.
3 . The recombinant fusion protein of claim 1 or 2 , wherein the hFSH alpha subunit and the hFSH beta subunit are independently linked to a terminus of the heavy chain constant 1 domain and a terminus of the light chain constant domain of the Fab.
4 . The recombinant fusion protein of any one of claims 1-3 , wherein the hFSH alpha subunit and the hFSH beta subunit are independently linked by a linker to a terminus of a heavy chain constant 1 domain and a terminus of a light chain constant domain of the Fab.
5 . The recombinant fusion protein of any one of claims 1-4 , wherein the hFSH alpha subunit is linked to a terminus of the light chain constant domain and the hFSH beta subunit is linked to a terminus of the heavy chain constant 1 domain of the Fab.
6 . The recombinant fusion protein of claim 4 or 5 , wherein the linker comprises 1 to 50 amino acids.
7 . The recombinant fusion protein of claim 6 , wherein the linker comprises an amino acid sequence of any one of SEQ ID NOS:16, 70-85, and 88.
8 . The recombinant fusion protein of any one of claims 1-7 , wherein the hFSH alpha subunit comprises an amino acid sequence having at least 90% identity to SEQ ID NO:1.
9 . The recombinant fusion protein of any one of claims 1-8 , wherein the hFSH alpha subunit comprises an amino acid sequence of SEQ ID NO:1.
10 . The recombinant fusion protein of any one of claims 1-9 , wherein the hFSH beta subunit comprises an amino acid sequence having at least 90% identity to SEQ ID NO:4.
11 . The recombinant fusion protein of any one of claims 1-10 , wherein the hFSH beta subunit comprises an amino acid sequence of SEQ ID NO:4.
12 . The recombinant fusion protein of any one of claims 1-11 , wherein the Fab comprises a heavy chain comprising a heavy chain variable domain comprising (a) a heavy chain complementarity determining domain 1 (CDR1) comprising the amino acid sequence of SYGIS (SEQ ID NO:22), a heavy chain complementarity determining domain 2 (CDR2) comprising the amino acid sequence of WINTYSGGTKYAQKFQG (SEQ ID NO:23), and a heavy chain complementarity determining domain 3 (CDR3) comprising the amino acid sequence of LGHCQRGICSDALDT (SEQ ID NO:24); (b) a heavy chain CDR1 comprising the amino acid sequence of SYGIS (SEQ ID NO:22), a heavy chain CDR2 comprising the amino acid sequence of RINTYNGNTGYAQRLQG (SEQ ID NO:25), and a heavy chain CDR3 comprising the amino acid sequence of LGHCQRGICSDALDT (SEQ ID NO:24); (c) a heavy chain CDR1 comprising the amino acid sequence of NYGIH (SEQ ID NO:26), a heavy chain CDR2 comprising the amino acid sequence of SISYDGSNKYYADSVKG (SEQ ID NO:27), and a heavy chain CDR3 comprising the amino acid sequence of DVHYYGSGSYYNAFDI (SEQ ID NO:28); (d) a heavy chain CDR1 comprising the amino acid sequence of SYAMS (SEQ ID NO:29), a heavy chain CDR2 comprising the amino acid sequence of VISHDGGFQYYADSVKG (SEQ ID NO:30), and a heavy chain CDR3 comprising the amino acid sequence of AGWLRQYGMDV (SEQ ID NO:31); (e) a heavy chain CDR1 comprising the amino acid sequence of AYWIA (SEQ ID NO:32), a heavy chain CDR2 comprising the amino acid sequence of MIWPPDADARYSPSFQG (SEQ ID NO:33), and a heavy chain CDR3 comprising the amino acid sequence of LYSGSYSP (SEQ ID NO:34); or (f) a heavy chain CDR1 comprising the amino acid sequence of AYSMN (SEQ ID NO:35), a heavy chain CDR2 comprising the amino acid sequence of SISSSGRYIHYADSVKG (SEQ ID NO:36), and a heavy chain CDR3 comprising the amino acid sequence of ETVMAGKALDY (SEQ ID NO:37); and a light chain comprising a light chain variable domain comprising (g) a light chain CDR1 comprising the amino acid sequence of RASQSISRYLN (SEQ ID NO:38), a light chain CDR2 comprising the amino acid sequence of GASRLES (SEQ ID NO:39), and a light chain CDR3 comprising the amino acid sequence of QQSDSVPVT (SEQ ID NO:40); (h) a light chain CDR1 comprising the amino acid sequence of RASQSISSYLN (SEQ ID NO:41), a light chain CDR2 comprising the amino acid sequence of AASSLQS (SEQ ID NO:42), and a light chain CDR3 comprising the amino acid sequence of QQSYSTPPYT (SEQ ID NO:43); (i) a light chain CDR1 comprising the amino acid sequence of RASQSIFNYVA (SEQ ID NO:44), a light chain CDR2 comprising the amino acid sequence of DASNRAT (SEQ ID NO:45), and a light chain CDR3 comprising the amino acid sequence of QQRSKWPPTWT (SEQ ID NO:46); (j) a light chain CDR1 comprising the amino acid sequence of RASETVSSRQLA (SEQ ID NO:47), a light chain CDR2 comprising the amino acid sequence of GASSRAT (SEQ ID NO:48), and a light chain CDR3 comprising the amino acid sequence of QQYGSSPRT (SEQ ID NO:49); (k) a light chain CDR1 comprising the amino acid sequence of RASQSVSSSSLA (SEQ ID NO:50), a light chain CDR2 comprising the amino acid sequence of GASSRAT (SEQ ID NO:48), and a light chain CDR3 comprising the amino acid sequence of QKYSSYPLT (SEQ ID NO:51); or (1) a light chain CDR1 comprising the amino acid sequence of RASQSVGSNLA (SEQ ID NO:52), a light chain CDR2 comprising the amino acid sequence of GASTGAT (SEQ ID NO:53), and a light chain CDR3 comprising the amino acid sequence of QQYYSFLAKT (SEQ ID NO:54).
13 . The recombinant fusion protein of any one of claims 1-12 , wherein the heavy chain variable domain comprises a heavy chain CDR1 comprising the amino acid sequence of SEQ ID NO:35, a heavy chain CDR2 comprising the amino acid sequence of SEQ ID NO:36, and a heavy chain CDR3 comprising the amino acid sequence of SEQ ID NO:37, and wherein the light chain variable domain comprises a light chain CDR1 comprising the amino acid sequence of SEQ ID NO:52, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO:53, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO:54.
14 . The recombinant fusion protein of any one of claims 1-13 , wherein the heavy chain variable domain comprises an amino acid sequence having at least 90% identity to SEQ ID NO:55, 56, 57, 58, 59, or 60.
15 . The recombinant fusion protein of any one of claims 1-14 , wherein the heavy chain variable domain comprises the amino acid sequence of SEQ ID NO:60.
16 . The recombinant fusion protein of any one of claims 1-15 , wherein the light chain variable domain comprises an amino acid sequence having at least 90% identity to SEQ ID NO:61, 62, 63, 64, 65, 66, or 67.
17 . The recombinant fusion protein of any one of claims 1-16 , wherein the light chain variable domain comprises the amino acid sequence of SEQ ID NO:67.
18 . The recombinant fusion protein of any one of claims 1-17 , wherein the heavy chain variable domain comprises an amino acid sequence of SEQ ID NO:55, 56, 57, 58, 59, or 60, and the light chain variable domain comprises an amino acid sequence of SEQ ID NO:61, 62, 63, 64, 65, 66, or 67.
19 . The recombinant fusion protein of any one of claims 1-18 , wherein the heavy chain constant domain comprises an amino acid sequence having at least 90% identity to SEQ ID NO:68.
20 . The recombinant fusion protein of any one of claims 1-19 , wherein the light chain constant domain comprises an amino acid sequence having at least 90% identity to SEQ ID NO:69 or 91.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority under 35 U.S.C. § 119 to Korean Appl. No. 10-2022-0126516, filed Oct. 4, 2022, the disclosure of which is incorporated by reference herein in its entirety. REFERENCE TO SEQUENCE LISTING SUBMITTED ELECTRONICALLY This application contains a Sequence Listing which has been submitted in XML format via EFS-Web and is hereby incorporated by reference in its entirety. Said XML copy, created on Oct. 4, 2023, is named 2662-0007WO01_SEQL_ST26 and is 130,077 bytes in size. BACKGROUND 1. Field The disclosure relates to a fusion protein including a follicle stimulating hormone and an antigen-binding fragment to serum albumin and uses thereof. 2. Description of the Related Art Infertility refers to a condition in which pregnancy does not happen even after being in a normal conjugal relationship for more than one year. The causes of infertility are estimated to lie in the male in 40%, the female in 40%, both the male and female in 10%, and unknown causes in 10%. In the case of female infertility, main causes include ovulation disorders, hormonal abnormalities, uterine abnormalities, and the like. In the case of male infertility, main causes include hormonal abnormalities, testicular abnormalities due to toxic substances or trauma, sperm abnormalities, and the like. Assisted reproductive technologies in the female infertility treatment include in vitro fertilization-embryo transfer (IVF-ET), gamete intrafallopian transfer (GIFT), zygote intrafallopian transfer (ZIFT), intracytoplasmic sperm injection (ICSI), and the like, such assisted reproductive technologies essentially include ovulation induction processes using follicle stimulating hormone. In addition, follicle stimulating hormone is used for therapeutic purposes in women suffering from anovulation. Furthermore, follicle stimulating hormone is also used for the treatment of male infertility caused by sperm abnormalities. Meanwhile, human follicle stimulating hormone (hFSH) is glycoprotein hormone produced by the pituitary gland and secreted in the endocrine system and has a dimer structure including an alpha subunit and a beta subunit. The alpha subunit belongs to a glycoprotein composed of 92 amino acids common to the glycoprotein families, such as luteinizing hormone (LH) as a pituitary glycoprotein, human chorionic gonadotropin (hCG) as a placental glycoprotein, and thyroid stimulating hormone (TSH), and has two N-glycosylation sites at positions 52 and 78 of asparagine. The beta subunit is a glycoprotein composed of 111 amino acids specific to hFSH and has two N-glycosylation sites at positions 7 and 24 of asparagine. The hFSH is a glycoprotein in which sugar chains account for about 40% of the total molecular weight, and such glycosylation is known to play an important role in the receptor binding ability and signal transduction activity of hFSH. The hFSH is known to play an important role in the differentiation and growth of male and female reproductive cells, and thus is used to mature multiple follicles in women who have undergone assisted reproductive technology for the infertility treatment or has been used for the treatment of anovulation. The hFSH is also used for sperm production and sperm count increase in men. Urine-derived hFSH purified from female urine has low efficiency and supply instability, and poses many risks, especially in terms of safety. In this regard, genetically engineered formulations prepared by using genetic engineering technology are being in common use, but due to a short in vivo half-life, there is a limitation that frequent administration of the genetically engineered formulations is required. Under such a technical background, there is a need to develop a long-acting hormone formulation as a technique for improving therapeutic efficacy of hFSH and increasing convenience of patients. Antigen-binding fragments that bind to serum albumin are provided in U.S. Pat. Nos. 9,879,077 and 11,773,176, each incorporated herein by reference in its entirety. SUMMARY Provided is a recombinant fusion protein including an antigen-binding fragment that binds to serum albumin and a human follicle stimulating hormone (hFSH). Provided are a nucleic acid encoding the recombinant fusion protein, an expression vector including the nucleic acid, and a cell transformed with the expression vector. Provided are a long-acting hFSH formulation or a pharmaceutical composition for infertility treatment, each including the recombinant fusion protein as an active ingredient. Additional aspects will be set forth in part in the description which follows and, in part, will be apparent from the description, or can be learned by practice of the presented embodiments of the disclosure. Disclosed herein are recombinant fusion proteins comprising (a) a human follicle stimulating hormone (hFSH) comprising an hFSH alpha subunit and an hFSH beta subunit and (b) an antigen binding fragment (Fab) that bi