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CN-122029185-A - TNF receptor 2 specific agonist fusion proteins comprising a TNF homology domain and a mutated Fc domain

CN122029185ACN 122029185 ACN122029185 ACN 122029185ACN-122029185-A

Abstract

The present disclosure relates to tumor necrosis factor receptor 2 (TNFR 2) agonist polypeptides comprising (i) a TNFR2 binding domain comprising three TNF Homology Domains (THDs) that specifically bind TNFR2, and (ii) an Fc domain. The present disclosure also relates to TNFR2 agonists for use in the treatment and/or prevention of, for example, chronic pain or multiple sclerosis.

Inventors

  • R. FISHER
  • U. Sarin
  • HANNA CHARLES
  • J. Bessia
  • ROLAND KONTERMANN
  • C. Vaslan
  • K. Pfeizemel
  • C. Terman
  • P. Prazon

Assignees

  • 雷萨诺有限责任公司
  • 百欧恩泰欧洲股份公司
  • 斯图加特大学

Dates

Publication Date
20260512
Application Date
20240927
Priority Date
20230928

Claims (16)

  1. 1. A tumor necrosis factor receptor 2 (TNFR 2) agonist polypeptide comprising (i) a TNFR2 binding domain comprising three TNF-homologous domains (THDs) that specifically bind to TNFR2, and (ii) an Fc domain, wherein the Fc domain comprises the following mutations L234A, L235A, A327G, A S and P331S.
  2. 2. The TNFR2 agonist polypeptide of claim 1, wherein (a) the Fc domain comprises (i) NO proline at position 233 and/or (ii) glycine at position 236, and/or (b) the Fc domain comprises a sequence having at least 85% sequence identity to SEQ ID No. 1.
  3. 3. The TNFR2 agonist polypeptide of claim 1 or claim 2, wherein each THD comprises an amino acid sequence having at least 80% sequence identity to SEQ ID No. 2, optionally wherein each THD independently comprises at least one mutation or a set of mutations ：D139Y、D139F、D139E、D139N、D139T、D139S、E142Q、E142H、E142K、A141R/S143T、Q84N/T89S/A141S/E142A/S143D、Q84N/A141I/E142G/S143D、A141H/E142S/S143D、A141H/S143D、L25V/A141D/E142D/S143D、A141N/E142D/S143D、A141T/E142S/S143D、A141Q/E142D/S143D、A141T/E142D/S143D、A141D/E142G/S143D、A141D/S143D、A141K/E142D/S143T、A141R/E142T/S143D、A141R/S143T、E142D/S143D、E142N/S143、S91C/G144C、K61A、K61W、Q63K、Q63T、Q63Y、L71H、L71W、D139W、D139V、D139V/F140L/A141S、D139N/A141R、D139V/A141S、L25V、L25T、L25S、L25A、L25G、R27H、R27I、R27L、R28G、R28E、S143L、S143R、S143P S143T、S143A、Q145E、Q145N、E142D、E142N、E142S、E142G、A141R、A141S、A141T、A141H、A141K、A141F、A141D、A141G、A141N、A141P、A141Q、A141Y、A141V and a141W, preferably at least one mutation or a set of mutations selected from the group consisting of D139N and a141R, in the corresponding position of the sequence of SEQ ID No. 2.
  4. 4. The TNFR2 agonist polypeptide according to any one of the preceding claims, wherein (a) one or more THDs, preferably each THD, comprises or consists of SEQ ID No. 3 or an amino acid sequence having at least 90% sequence identity to SEQ ID No. 3, and/or (b) one THD comprises or consists of SEQ ID No. 4 or an amino acid sequence having at least 90% sequence identity to SEQ ID No. 4.
  5. 5. The TNFR2 agonist polypeptide of any one of the preceding claims, wherein the TNFR2 binding domain comprises or consists of a sequence having at least 80% sequence identity to SEQ ID No. 5.
  6. 6. The TNFR2 agonist polypeptide of any one of the preceding claims, wherein the TNFR2 binding domain is linked to the Fc domain by a flexible linker consisting of 3-20 amino acids, optionally wherein the flexible linker is a Ser-Gly linker, preferably GGSGGGGSGG (SEQ ID NO: 6).
  7. 7. The TNFR2 agonist polypeptide of any one of the preceding claims, wherein the TNFR2 agonist polypeptide comprises a sequence having at least 90% sequence identity to SEQ ID No. 7.
  8. 8. A TNFR2 agonist multimer, wherein the TNFR2 agonist multimer is in the form of a dimer comprising two TNFR2 agonist polypeptides, wherein each TNFR2 agonist polypeptide is independently a TNFR2 agonist polypeptide according to any preceding claim, optionally wherein each TNFR2 agonist polypeptide in the dimer is the same.
  9. 9. A polynucleotide encoding a TNFR2 agonist polypeptide according to any one of claims 1 to 7.
  10. 10. A vector comprising the polynucleotide according to claim 9.
  11. 11. A pharmaceutical composition comprising a TNFR2 agonist polypeptide according to any one of claims 1 to 7, a TNFR2 agonist multimer according to claim 8, a polynucleotide according to claim 9 or a vector according to claim 10.
  12. 12. A TNFR2 agonist comprising three TNF-homologous domains (THD) and an Fc domain that specifically bind to TNFR2 for use in the treatment and/or prevention of chronic pain or multiple sclerosis.
  13. 13. The TNFR2 agonist for use according to claim 12, wherein the TNFR2 agonist is a TNFR2 agonist polypeptide according to any one of claims 1 to 7, a TNFR2 agonist multimer according to claim 8, a polynucleotide according to claim 9, a vector according to claim 10, or provided as a pharmaceutical composition according to claim 11.
  14. 14. The TNFR2 agonist for use according to claim 12 or claim 13, wherein the TNFR2 agonist induces TNFR2 signaling in a neuron, optionally wherein the neuron is an on-spinal-cord neuron, preferably a nex+ on-spinal-cord neuron.
  15. 15. The TNFR2 agonist for use according to any one of claims 12 to 14, wherein the TNFR2 agonist is administered to a subject, wherein the administration is selected from the list consisting of systemic administration, intravenous administration, parenteral administration (INTRAPARENTERAL ADMINISTRATION), subcutaneous administration, intrathecal administration, and intra-articular administration.
  16. 16. The TNFR2 agonist for use according to any one of claims 11 to 15, wherein the chronic pain is associated with osteoarthritis.

Description

TNF receptor 2 specific agonist fusion proteins comprising a TNF homology domain and a mutated Fc domain FIELD The present disclosure relates to tumor necrosis factor receptor 2 (TNFR 2) agonist polypeptides comprising (i) a TNFR2 binding domain comprising three TNF Homology Domains (THDs) that specifically bind to TNFR2, and (ii) an Fc domain. The present disclosure also relates to TNFR2 agonists for use in the treatment and/or prevention of, for example, chronic pain (e.g., chronic neuropathic pain) or multiple sclerosis. Background The Tumor Necrosis Factor (TNF) superfamily is a family of structurally related cytokines that have a variety of functions. TNF itself is a multifunctional cytokine with pleiotropic functions. It is a major regulator of the immune system, and is also a key member of the initiation and coordination of inflammation and immunity. Like most ligands of this superfamily, TNF is synthesized as a trimeric transmembrane protein type 2 (tmTNF), which can be proteolytically processed into soluble circulating TNF homotrimers (sTNF). Interestingly, sTNF and tmTNF activate two different TNF receptors (TNFRs), TNFR1 and TNFR2, with different abilities. Although TNFR1 is activated by both sTNF and tmTNF, potent activation of TNFR2 is dependent on tmTNF (Muhlenbeck et al, 2000, J.biol., chem.275, 32208-32213; wajant et al, 2001, oncogene 20, 4101-4106). There remains a need for improved TNFR2 agonists. In particular, there remains a need for TNFR2 agonists with advantageous properties for in vivo administration and for use as a medicament. SUMMARY The present disclosure is based, at least in part, on the inventors' development of TNFR2 agonist polypeptides having particularly advantageous and surprising properties. For example, the inventors have developed TNFR2 agonist polypeptides that have reduced binding and activation to fcyriia and have beneficial in vivo properties. Furthermore, TNFR2 agonist polypeptides of the present invention have been shown to have significant efficacy in disease-related pathways, including direct effects on neurons on the spinal cord. Without wishing to be bound by theory, the inventors believe that the direct effect on neurons on the spinal cord-unlike the previously reported effect on Treg cells-may promote the long-term and prophylactic effects of TNFR2 agonists of the invention. Accordingly, the present disclosure provides tumor necrosis factor receptor 2 (TNFR 2) agonist polypeptides comprising (i) a TNFR2 binding domain comprising three TNF-homologous domains (TNF homology domain, THD) that specifically bind to TNFR2, and (ii) an Fc domain, wherein the Fc domain comprises the following mutations L234A, L235A, A327G, A S and P331S. Suitably, the Fc domain may not comprise a proline at position 233. Suitably, the Fc domain may comprise glycine at position 236. Suitably, the Fc domain (i) may not comprise a proline at position 233, and (ii) may comprise a glycine at position 236. Suitably, the Fc domain may comprise a sequence having at least 85% sequence identity to SEQ ID NO. 1. Suitably, each THD may independently comprise at least one mutation or at least one set of mutations ：D139Y、D139F、D139E、D139N、D139T、D139S、E142Q、E142H、E142K、A141R/S143T、Q84N/T89S/A141S/E142A/S143D、Q84N/A141I/E142G/S143D、A141H/E142S/S143D、A141H/S143D、L25V/A141D/E142D/S143D、A141N/E142D/S143D、A141T/E142S/S143D、A141Q/E142D/S143D、A141T/E142D/S143D、A141D/E142G/S143D、A141D/S143D、A141K/E142D/S143T、A141R/E142T/S143D、A141R/S143T、E142D/S143D、E142N/S143、S91C/G144C、K61A、K61W、Q63K、Q63T、Q63Y、L71H、L71W、D139W、D139V、D139V/F140L/A141S、D139N/A141R、D139V/A141S、L25V、L25T、L25S、L25A、L25G、R27H、R27I、R27L、R28G、R28E、S143L、S143R、S143P S143T、S143A、Q145E、Q145N、E142D、E142N、E142S、E142G、A141R、A141S、A141T、A141H、A141K、A141F、A141D、A141G、A141N、A141P、A141Q、A141Y、A141V and A141W selected from the group consisting of in the corresponding position of the sequence of SEQ ID NO: 2. It will be appreciated that the term "set of mutations" as used above refers to a combination of mutations denoted "/", wherein all mutations in the combination are selected, e.g., if "Q84N/T89S/A141S/E142A/S143D" as used above is selected, all denoted mutations will be present in the amino acid sequence. Suitably, each THD may comprise a sequence having at least 85% sequence identity to SEQ ID NO. 2. Suitably, THDs according to the present disclosure may comprise mutations D139N and A141R at positions corresponding to the sequence of SEQ ID NO: 2. Suitably, one or more THDs may comprise or consist of SEQ ID NO. 3 or an amino acid sequence having at least 95% sequence identity to SEQ ID NO. 3. Suitably, each THD may comprise or consist of SEQ ID NO. 3 or an amino acid sequence having at least 95% sequence identity with SEQ ID NO. 3. Suitably, one THD may comprise or consist of SEQ ID NO. 4 or an amino acid sequence having at least 95% sequence identity with SEQ ID NO. 4. Suitably, the first and second THDs can be directly linked without any intervenin