IP Library › Granted Patent US 12,454,577
Granted Patent B2
US 12,454,577 · App. 16/972,654 · Granted Oct 28, 2025

Trimeric polypeptide complexes comprising a collagen XVIII homotrimerization domain and/or a collagen xv homotrimerization and an agonist of a TNFR family costimulatory receptor, encoding polynucleotide thereof and method of use thereof to treat cancer

Inventors: Marta Compte Grau (Madrid, ES); Luis Álvarez Vallina (Madrid, ES)
Assignee: LEADARTIS, S.L.
C07K16/2878A61P35/00C07K16/2863C07K16/3007A61K2039/505C07K2317/31C07K2317/75C07K2317/92C07K2317/94
View Patent ↗
Loading inventors, assignments & file history…
Monitor This Case
Get email alerts when status or documents change.
Order Certified Copies
Most orders are placed with the USPTO same day — all within 24 business hours.
Order via The Patent Place →
Pre-filled with this patent's details
Quick Facts
Patent No.
US 12,454,577
App. No.
16/972,654
Granted
Oct 28, 2025
Kind
B2
Abstract

The invention relates to tumor-targeted trimeric polypeptides based on the homotrimerization region from collagen XV or XVIII and which contain domains capable of specifically binding to and activating receptors of the TNFR family of costimulatory receptors as well as to the uses thereof in the treatment of cancer.

Claims (40)

1. A trimeric polypeptide complex comprising three monomer polypeptides

wherein each monomer polypeptide comprises:

i. a homotrimerization domain selected from the group consisting of the collagen XVIII homotrimerization domain (TIE XVIII ), the collagen XV homotrimerization domain (TIE XV ) and a functionally equivalent variant thereof, wherein the functionally equivalent variant of TIE XVIII is a sequence showing at least 80% identity with the sequence SEQ ID NO: 12 and the functionally equivalent variant of TIE XV is a sequence showing at least 80% identity with the sequence SEQ ID NO: 2; and

ii. an agonist of a TNFR family costimulatory receptor, and

wherein at least one monomer polypeptide further comprises a polypeptide region which is capable of specifically binding to a tumor associated antigen.

2. The trimeric polypeptide complex according to claim 1 wherein the region which is capable of specifically binding to the tumor associated antigen is positioned N-terminal or C-terminal with respect to the homotrimerization domain.

3. The trimeric polypeptide complex according to claim 2 wherein

a. if the region which is capable of specifically binding to the tumor associated antigen is positioned N-terminal with respect to the homotrimerization domain, then the agonist of a TNFR family costimulatory receptor is positioned C-terminal with respect to the homotrimerization domain, or

b. if the region which is capable of specifically binding to the tumor associated antigen is positioned C-terminal with respect to the homotrimerization domain, then the agonist of a TNFR family costimulatory receptor is positioned N-terminal with respect to the homotrimerization domain.

4. The trimeric polypeptide complex according to claim 1 wherein the TNFR family costimulatory receptor is 4-1BB.

5. The trimeric polypeptide complex according to claim 1 wherein the agonist of the TNFR family costimulatory receptor is an agonistic antibody.

6. The trimeric polypeptide complex according to claim 1 wherein the tumor associated antigen is the epidermal growth factor receptor (EGFR).

7. The trimeric polypeptide complex according to claim 6 , wherein the polypeptide region which is capable of specifically binding to EGFR is an antibody.

8. The trimeric polypeptide complex according to claim 7 , wherein the anti-EGFR antibody is the EGA1 nanobody encoded by the sequence shown in SEQ ID NO: 3.

9. The trimeric polypeptide complex according to claim 1 wherein the tumor associated antigen is a carcinoembryonic antigen (CEA).

10. The trimeric polypeptide complex according to claim 9 , wherein the polypeptide region which is capable of specifically binding to CEA is an anti-CEA antibody.

11. The trimeric polypeptide complex according to claim 1 wherein the agonist of the TNFR family costimulatory receptor, the polypeptide region which is capable of specifically binding to the tumor associated antigen and/or the homotrimerization domain are either directly linked or linked through a spacer.

12. The trimeric polypeptide complex according to claim 1 wherein the agonist of the TNFR family costimulatory receptor is linked to the homotrimerization domain through a 18-residue-long linker and/or wherein the polypeptide region which is capable of specifically binding to the tumor associated antigen is linked to homotrimerization domain through a 16-residue long linker.

13. The trimeric polypeptide complex according to claim 1 wherein at least one of the monomers further comprises a tag suitable for detection and/or purification of the trimeric polypeptide complex and/or wherein at least one of the monomers further comprise a moiety which increases its circulation half-life.

14. The trimeric polypeptide complex according to claim 5 wherein the agonistic antibody of the TNFR family costimulatory receptor is a scFv, a nanobody or an antibody mimetic.

15. The trimeric polypeptide complex according to claim 6 , wherein the polypeptide region which is capable of specifically binding to EGFR is a scFv, a nanobody or an antibody mimetic.

16. The trimeric polypeptide complex according to claim 10 , wherein the anti-CEA antibody is a scFv, a nanobody or an antibody mimetic.

17. The trimeric polypeptide complex according to claim 11 wherein the spacer is a flexible linker with between 1 and 18 residues.

18. The trimeric polypeptide complex according to claim 13 wherein the moiety that increases the circulation half-life of the trimer is an albumin fragment or an albumin-binding moiety.

19. A pharmaceutical composition comprising a trimeric polypeptide complex according to claim 1 .

20. A polynucleotide encoding the monomer polypeptide forming part of the trimeric polypeptide complex as defined in claim 1 , wherein the monomer polypeptide comprises:

a homotrimerization domain selected from the group consisting of the collagen XVIII homotrimerization domain (TIE XVIII ), the collagen XV homotrimerization domain (TIE XV ) and a functionally equivalent variant thereof; and

an agonist of a TNFR family costimulatory receptor, and

a polypeptide region which is capable of specifically binding to a tumor associated antigen.

21. A vector comprising the polynucleotide according to claim 20 .

22. A host cell comprising a vector according to claim 21 .

23. A method for producing a trimeric polypeptide complex according to claim 1 which comprises isolating said trimeric polypeptide complex from a culture comprising a host cell which carries and expresses a polynucleotide which encodes a polypeptide comprising:

a homotrimerization domain selected from the group consisting of the collagen XVIII homotrimerization domain (TIE XVIII ), the collagen XV homotrimerization domain (TIE XV ) and a functionally equivalent variant thereof; and

an agonist of a TNFR family costimulatory receptor, and

a polypeptide region which is capable of specifically binding to a tumor associated antigen,

and optionally subjecting the trimeric polypeptide to further processing.

24. A method of treating cancer which comprises administering the trimeric polypeptide complex according to claim 1 to a subject in need thereof.

25. The method for treating cancer according to claim 24 , wherein the cancer is positive for the tumor associated antigen which is specifically recognized by the polypeptide region capable of specifically binding to a tumor associated antigen present in the trimeric polypeptide complex.

26. The method for treating cancer according to claim 25 , wherein the cancer is selected from the group consisting of colorectal cancer, lung cancer, breast cancer, pancreatic, renal, head and neck, gastric cancer, esophageal cancer, gynecologic cancer, prostate cancer, urothelial, a neurological cancer, and an haematological cancer.

27. The method according to claim 26 , wherein the gynecologic cancer is selected from the group consisting of ovarian, cervical and endometrial cancer, the neurological cancer is glioblastoma multiforme or the haematological cancer is acute myelogenous leukaemia.

Priority Claims (1)
EP 18382401 · Jun 6, 2018 · regional
Continuity (1)
Related Publication 20210246217A1 · Aug 12, 2021
References Cited (38)
US 11485790B2 · Igawa et al. · 2022 [cited by applicant]
US 20150139991A1 · Alvarez et al. · 2015 [cited by applicant]
US 20190169308A1 · Dahlen et al. · 2019 [cited by applicant]
WO WO9418227 · 1994 [cited by applicant]
WO 2012049328A1 · 2012 [cited by applicant]
WO WO2017098005 · 2017 [cited by applicant]
WO 2017182672A1 · 2017 [cited by applicant]
Cuningham BC and Wells JA. (Jun. 2, 1989) Science. 244:1081-1085. (DOI: 10.1126/science.2471267). [cited by examiner]
Pak MA, et al. (2023) PLoS ONE. 18(3):9 pages. e0282689. (https://doi.org/10.1371/journal.pone.0282689). [cited by examiner]
Melero, I. et al., Monoclonal Antibodies against the 4-1BB T-cell activation molecule eradicate established tumors, 1997, Nature Medicine, vol. 3, No. 6, pp. 682-685. [cited by applicant]
Neil H. Segal et al., Results from an Integrated Safety Analysis of Urelumab, an Agonist Anti-CD137 Monoclonal Antibody, 2016 Clinical Cancer Research, vol. 23, No. 8, pp. 1929-1936. [cited by applicant]
Altschul et al. Gapped BLAST and PSI-BLAST: a new generation of protein database search programs, [cited by applicant]
Binz et al. Engineering novel binding proteins from nonimmunoglobulin domains, [cited by applicant]
Chothia et al. Canonical Structures for the Hypervariable Regions of Immunoglobulins, [cited by applicant]
Henikoff et al. Amino acid substitution matrices from protein blocks, [cited by applicant]
Humphrey et al. Anti-synthetic peptide antibody reacting at the fusion junction of deletion-mutant epidermal growth factor receptors in human glioblastoma, [cited by applicant]
James at al. Benzodiazepine Peptidomimetics: Potent Inhibitors of Ras Farnesylation in Animal Cells, [cited by applicant]
Jones. Proteinase Mutants of [cited by applicant]
Lefranc et al. IMGT unique numbering for immunoglobulin and T cell receptor variable domains and Ig superfamily V-like domains, [cited by applicant]
Niu et al. Cytokine-Mediated Disruption of Lymphocyte Trafficking, Hemopoiesis, and Induction of Lymphopenia, Anemia, and Thrombocytopenia in Anti-CD137-Treated Mice, [cited by applicant]
Wu et al. An Analysis of the Sequences of the Variable Regions of Bence Jones Proteins and Myeloma Light Chains and their Implications for Anti- Body Complementarity, [cited by applicant]
International Search Report, International Patent Application No. PCT/EP2019/064864, Aug. 14, 2019. [cited by applicant]
Blanco-Toribio et al.; “Generation and characterization of monospecific and bispecific hexavalent trimerbodies,” 2013 MAbs. vol. 5(1): pp. 70-79. [cited by applicant]
Harwood et al.; “ATTACK, a novel bispecific T cell-recruiting antibody with trivalent EGFR binding and monovalent CD3 binding for cancer immunotherapy,” 2018 Oncoimmunology, vol. 7(1):e1377874; https://doi.org/10.1080/2… [cited by applicant]
Hinner et al.; “Costimulatory T cell engagement via a novel bispecific anti-CD137 /anti-HER2 protein,” 2015, Journal for Immuno Therapy of Cancer, vol. 3(Suppl 2):P187; doi:10.1186/2051-1426-3-S2-P187. [cited by applicant]
Hinner et al., “Costimulatory T cell engagement by PRS-343, a 4-1BB (CD137)/HER2 bispecific, leads to tumor growth inhibition and TIL expansion in humanized mouse model,” 2016, ABSTRACT. [cited by applicant]
Shuford et al.; “4-1BB Costimulatory Signals Preferentially Induce CD8+ T Cell Proliferation and Lead to Amplification In Vivo of Cytotoxic T Cell Responses,” 1997, J Exp Med, vol. 186(1): pp. 47-55. [cited by applicant]
Schmitz et al.; “Structural evaluation of EGFR inhibition mechanisms for nanobodies/VHH domains,” 2013, Structure vol. 21(7): pp. 1214-1224. [cited by applicant]
Compte et al.; “A tumor-targeted trimeric 4-1BB-agonistic antibody induces potent anti-tumor immunity without systemic toxicity,” 2018, Nat Commun, vol. 9(1):4809; DOI: 10.1038/s41467-018-07195-w. [cited by applicant]
Bartkowiak et al.; “4-1BB agonists: multi-potent potentiators of tumor immunity,” 2015, Front Oncol, vol. 5:117; doi: 10.3389/fonc.2015.00117. [cited by applicant]
Alvarez-Cienfuegos et al. “Intramolecular trimerization, a novel strategy for making multispecific antibodies with controlled orientation of the antigen binding domains,” 2016, Sci Rep, vol. 6:28643; DOI: 10.1038/srep28… [cited by applicant]
Cuesta et al.; “In Vivo Tumor Targeting and Imaging with Engineered Trivalent Antibody Fragments Containing Collagen-Derived Sequences,” 2009, PLoS On, e 4(4):e5381; doi:10.1371/journal.pone.0005381. [cited by applicant]
Compte et al., An Fc-free EGFR-specific 4-1BB-agonistic Trimerbody Displays Broad Antitumor Activity in Humanized Murine Cancer Models without Toxicity, Jun. 1, 2021, Clinical Cancer Research, vol. 27, No. 11, pp. 3167-… [cited by applicant]
Mittler et al., Anti-4-1BB Monoclonal Antibodies Abrogate T Cell-dependent Humoral Immune Responses In Vivo through the Induction of Helper T Cell Anergy, Nov. 15, 1999, Journal of Experimental Medicine, vol. 190, No. 1… [cited by applicant]
Muik et al., Preclinical Characterization and Phase I Trial Results of a Bispecific Antibody Targeting PD-L1 and 4-1BB (GEN1046) in Patients with Advanced Refractory Solid Tumors, May 2022, Cancer Discovery, vol. 12, pp… [cited by applicant]
Wu et al., Phase I study of the efficacy and safety of IB1319 in patients with advanced malignant tumors, 2022, Journal of Clinical Oncology, vol. 40, No. 16 Suppl: 2646. [cited by applicant]
Compte et al., An EGFR-Targeted 4-1BB-agonistic Trimerbody Does Not Induce Hepatotoxicity in Transgenic Mice With Liver Expression of Human EGFR, Frontiers in Immunology, Jan. 7, 2021, vol. 11, art. 614363. [cited by applicant]
Silva-Pilipich et al., Local delivery of optimized nanobodies targeting the PD-1/PD-L1 axis with a self-amplifying RNA viral vector induces potent antitumor responses, Cancer Letters, May 1, 2023, vol. 561, art. 216139. [cited by applicant]