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JP-2026514390-A - GnRH-binding polypeptides and their use

JP2026514390AJP 2026514390 AJP2026514390 AJP 2026514390AJP-2026514390-A

Abstract

This invention relates to the field of GnRH antagonists and anti-GnRH ligands. The invention also relates to the field of pharmaceuticals and diagnostics. In particular, this disclosure relates to the field of contraception and GnRH-related diseases.

Inventors

  • デルコンベル，ロマン
  • オーウェン，チャールズ
  • ピメンタ・デ・カストロ，イネス
  • ベルテッリ，フランソワーズ

Assignees

  • ２７２バイオ・リミテッド
  • ビオアズ

Dates

Publication Date
20260511
Application Date
20240329
Priority Date
20230330

Claims (15)

  1. An isolated monoclonal anti-gonadotropin-releasing hormone (anti-GnRH) single-domain antibody (SdAb) or its antigen-binding fragment, which specifically binds to both the N-terminal region (pGlu-His-Trp) and the C-terminal region (Pro-Gly- NH2 ) of native GnRH.
  2. An isolated monoclonal anti-gonadotropin-releasing hormone (anti-GnRH) antibody or its antigen-binding fragment, - CDR3 polypeptide sequence of sequence number 33 or 188; - CDR3 polypeptide sequence of sequence number 57 or 187; - CDR3 polypeptide sequence of sequence number 36; - CDR3 polypeptide sequence of sequence number 96; - CDR3 polypeptide sequence of sequence number 48; - CDR3 polypeptide sequences of sequence numbers 42, 75, 81, 90, and 93; or - An isolated antibody or its antigen-binding fragment comprising a variable domain having a complementarity-determining region 3 (CDR3) selected from the group consisting of variants of the CDR3 polypeptide sequence having one or two substitutions.
  3. The isolated antibody or antigen-binding fragment thereof according to claim 2, wherein the antibody is a single-domain antibody (SdAb).
  4. An isolated antibody or antigen-binding fragment thereof according to any one of claims 1 to 3, characterized by comprising a variable domain having at least one complementarity-determining region 3 (CDR3) polypeptide sequence of SEQ ID NO: 57 or 187.
  5. - CDR1 containing sequences selected from sequence numbers 55 and 82; - CDR2 containing sequences selected from sequence numbers 56, 98, 116, and 199; - CDR3 containing the sequence of sequence number 57 An isolated antibody or antigen-binding fragment thereof according to claim 1 or 2, characterized by comprising a variable domain having a variable domain.
  6. - CDR1 containing sequences selected from sequence numbers 31 and 195; - CDR2 containing the sequence selected from sequence number 32; - CDR3 containing the sequence of sequence number 33 An isolated antibody or antigen-binding fragment thereof according to claim 1 or 2, characterized by comprising a variable domain having a variable domain.
  7. - CDR1 containing the sequence selected from sequence number 34, CDR2 containing the sequence selected from sequence number 35, CDR3 containing the sequence of sequence number 36; or - CDR1 containing the sequence selected from SEQ ID NO: 94, CDR2 containing the sequence selected from SEQ ID NO: 95, CDR3 containing the sequence of SEQ ID NO: 96; or - CDR1 containing the sequence selected from sequence number 46, CDR2 containing the sequence selected from sequence number 47, and CDR3 containing the sequence from sequence number 48. An isolated antibody or antigen-binding fragment thereof according to claim 1 or 2, characterized by comprising a variable domain having a variable domain.
  8. An isolated antibody or antigen-binding fragment thereof according to any one of claims 1 to 7, wherein the GnRH is selected from the group consisting of human GnRH, porcine GnRH, bovine GnRH, horse GnRH, sheep GnRH, dog GnRH, and feline GnRH.
  9. A nucleic acid comprising an antibody or its antigen-binding fragment according to any one of claims 1 to 8.
  10. A vector comprising the nucleic acid described in claim 9.
  11. A host cell comprising the nucleic acid described in claim 9 or the vector described in claim 10.
  12. - An isolated antibody or antigen-binding fragment thereof according to any one of claims 1 to 8, or a nucleic acid according to claim 9, or a vector according to claim 10, or a host cell according to claim 11; and - A pharmaceutical composition containing pharmaceutically acceptable excipients.
  13. For use as a pharmaceutical or for use in an in vivo diagnostic method, an isolated antibody or its antigen-binding fragment according to any one of claims 1 to 8, or a nucleic acid according to claim 9, or a vector according to claim 10, or a host cell according to claim 11, or a pharmaceutical composition according to claim 12.
  14. An in vitro method for detecting gonadotropin-releasing hormone (GnRH), a) A step of providing a sample, particularly a biological sample or a fraction thereof; b) A method comprising the step of contacting a sample with an isolated antibody or its antigen-binding fragment according to any one of claims 1 to 8.
  15. A method for isolating an anti-gonadotropin-releasing hormone (anti-GnRH) antibody or its antigen-binding fragment, a) Providing a library of single-domain antibodies (SdAbs) or fragments thereof; b) A method comprising the step of subjecting the library to antigen affinity selection, wherein the selected SdAb or fragment thereof has specificity for both the N-terminal region of natural GnRH (pGlu-His-Trp) and the C-terminal region of natural GnRH (Pro-Gly- NH2 ).

Description

This invention relates to the field of GnRH antagonists and anti-GnRH ligands. This invention also relates to the fields of pharmaceuticals and diagnostics. In particular, this disclosure relates to the fields of contraception and GnRH-related disorders. Various methods of contraception are being implemented in this field, particularly for livestock, including pigs. The long-established method of sterilization for pigs consisted of live castration of piglets without any anesthesia. However, this long-standing method has been considered unethical due to the unacceptable pain, intense suffering, and distress it inflicts on the piglets. Methods of animal contraception known in the art also include surgical contraception under anesthesia (e.g., tubal ligation or vasectomy), hormone-containing implants for insertion of intrauterine contraceptive devices (e.g., GnRH analog-containing implants), the use of hormone-containing compositions including direct injection of chemicals into the testes to destroy reproductive tissue, and methods for immunological contraception. GnRH-binding ligands, particularly monoclonal antibodies, have been reported in this field. Known immunological methods of contraception include the administration of vaccine compositions aimed at immunizing the target against endogenous GnRH. In particular, GnRH vaccines stimulate antibody production, which in turn inactivates endogenous GnRH, leading to a decrease in gonadotropin release that results in gonad atrophy in adult animals or stunted growth in sexually immature animals. The resulting regression or lack of development of the reproductive organs is called immunological sterilization. Because they function as hormones, GnRH, GnRH analogs, and GnRH-binding ligands have also been used as therapeutic agents in numerous other medical conditions and clinical applications, including the treatment or prevention of diseases beyond infertility, particularly sex hormone-dependent disorders such as delayed puberty, ovarian hyperstimulation syndrome, precocious puberty, premenstrual syndrome, endometriosis, uterine fibroids, polycystic ovary syndrome, hirsutism, acne vulgaris, acute intermittent porphyria, breast cancer, uterine cancer, uterine fibroids, endometrial cancer, prostate cancer, benign prostatic hyperplasia, and pancreatic cancer, as either GnRH agonists or GnRH antagonists. Silversides et al. ("Monoclonal antibodies against LHRH: development and immunoactivity in vivo and in vitro"; Journal of Reproductive Immunology, 1985) reported monoclonal antibodies derived from canine immunoassays against GnRH/LHRH. Talwar et al. ("Bioeffective monoclonal antibody against the decapeptide gonadotropin-releasing hormone: Reacting determinant and action on ovulation and estrus suppression," Proc. Natl. Acad. Sci. USA, 1985) reported an antibody preparation capable of suppressing the progression of estrus in dogs. Li et al. ("Vectorized antibody gene delivery mediates long-term contraception," Current Biology, 2015) and US10,570,200B2 teach antibody-mediated immunocontraception, including vectorized AAV-dependent anti-GnRH antibody expression. A purified monoclonal mouse IgG1, kappa antibody, specifically targeting the C-terminal peptide of GnRH and known as clone number SMI41 (absolute antibody reference number Ab00922-1.1), has also been reported for immunocytochemistry and expressed in mice via injection of non-replicating recombinant adeno-associated virus to induce prolonged infertility. CEVA VALORA (trademark) is reported as a GnRH/LHRH-based synthetic peptide vaccine for pigs, containing three synthetic peptides as immunogens. IMPROVAC® has been reported as a GnRH analog protein conjugate capable of stimulating the porcine immune system to produce antibodies against GnRH, thereby controlling sexual development. Chang et al. ("Effects of a recombinant gonadotropin-releasing hormone vaccine on reproductive function in adult male ICR mice"; Vaccines, 2021) reported a vaccine preparation containing a multimeric fusion protein that includes multiple copies of the GnRH I decapeptide and T cell epitope. Gonadotropin-releasing hormone (GnRH), also known as LHRH, is a well-conserved hormone in mammals, originating in the central hypothalamus. Secreted by the pituitary gland, it acts as a major reproductive hormone by regulating the release of two main gonadotropins: luteinizing hormone (LH) and follicle-stimulating hormone (FSH). GnRH has been extensively studied for the development of immunizing contraceptive vaccines for various mammalian species, including pigs, rodents, cats, dogs, and wild carnivores. The amino acid sequence of GnRH is highly conserved among phylogenetically closely related species; it is generally reported as a decapeptide characterized by a conserved N-terminal region, a central region, and a conserved C-terminal region, with the following amino acid structure provided for reference in mammals: (Pyro)Glu-His-Trp-Ser-Tyr-Gly-Leu-Arg-