IP Library › Granted Patent US 12,448,447
Granted Patent B2
US 12,448,447 · App. 18/060,694 · Granted Oct 21, 2025

CD8 binding agents

Inventors: Nikolai Kley (Newton, MA); Jan Tavernier (Balegem, BE); Lennart Zabeau (Ghent, BE); Erik Depla (Ghent, BE)
Assignees: Orionis Biosciences, Inc.; Orionis Biosciences BV
C07K16/2815C07K14/56C07K2317/22C07K2317/24C07K2317/565C07K2317/569C07K2319/02
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Quick Facts
Patent No.
US 12,448,447
App. No.
18/060,694
Granted
Oct 21, 2025
Kind
B2
Abstract

The present invention relates, in part, to agents that bind CD8 and their use as therapeutic and diagnostic agents. The present invention further relates to pharmaceutical compositions comprising the CD8 binding agents and their use in the treatment of various diseases, including, for example, cancers.

Claims (16)

1. A chimeric protein comprising a CD8 binding agent comprising:

(a) a targeting moiety having (i) an amino acid sequence having at least 90% identity with SEQ ID NO: 1217 and having a CDR3 consisting of the amino acid sequence of SEQ ID NO: 166; (ii) an amino acid sequence having at least 90% identity with SEQ ID NO: 1218 and having a CDR3 consisting of the amino acid sequence of SEQ ID NO: 177; (ii) an amino acid sequence having at least 90% identity with SEQ ID NO: 1219 and having a CDR3 consisting of the amino acid sequence of SEQ ID NO: 181; (iv) an amino acid sequence having at least 90% identity with SEQ ID NO: 1220 and having a CDR3 consisting of the amino acid sequence of SEQ ID NO: 199; or (v) an amino acid sequence having at least 90% identity with SEQ ID NO: 1216 and having a CDR3 consisting of the amino acid sequence of SEQ ID NO: 183, and

(b) a modified human interferon alpha 2 (IFN-α2) molecule having the amino acid sequence of SEQ ID NO: 288 or 289 and comprising a R149A mutation, relative to SEQ ID NO: 288 or 289, that confers reduced affinity or activity for its receptor, wherein the reduced affinity or activity at the receptor is restorable by attachment with the CD8 binding agent.

2. The chimeric protein of claim 1 , further comprising one or more additional targeting moieties.

3. The chimeric protein of claim 2 , wherein the one or more additional targeting moieties recognize a tumor antigen.

4. The chimeric protein of claim 2 , wherein the one or more additional targeting moieties modulate a tumor antigen.

5. The chimeric protein of claim 2 , wherein the one or more additional targeting moieties recognize an antigen on an immune cell.

6. The chimeric protein of claim 2 , wherein the one or more additional targeting moieties modulate an antigen on an immune cell.

7. The chimeric protein of claim 6 , wherein the immune cell is selected from a T cell, a B cell, a dendritic cell, a macrophage, neutrophil, and a NK cell.

8. The chimeric protein of claim 1 , wherein the chimeric CD8 binding agent recruits cytotoxic T cells to tumor cells or to the tumor environment.

9. The chimeric protein of claim 1 , wherein the modified human IFN-α2 further comprises one or more mutations selected from R120E, A145G, M148A, and L153A, relative to SEQ ID NO: 288 or 289.

10. The chimeric protein of claim 1 , wherein the targeting moiety has an amino acid sequence having at least 90% identity with SEQ ID NO: 1217 and having a CDR3 consisting of the amino acid sequence of SEQ ID NO: 166.

11. The chimeric protein of claim 1 , wherein the targeting moiety has an amino acid sequence having at least 92% identity with SEQ ID NO: 1217.

12. A recombinant nucleic acid encoding the chimeric protein of claim 1 .

13. A host cell comprising the recombinant nucleic acid of claim 12 .

14. A method for treating or preventing cancer, comprising administering to a patient in need thereof an effective amount of the chimeric protein of claim 1 .

Assignments (3)
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Feb 27, 2023
From: KLEY, NIKOLAI
To: ORIONIS BIOSCIENCES, INC.
Reel/Frame 062806/0436 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Feb 27, 2023
From: TAVERNIER, JAN; ZABEAU, LENNART; DEPLA, ERIK
To: ORIONIS BIOSCIENCES NV
Reel/Frame 062806/0442 →
CHANGE OF NAME Recorded Feb 27, 2023
From: ORIONIS BIOSCIENCES NV
To: ORIONIS BIOSCIENCES BV
Reel/Frame 062871/0754 →
Continuity (3)
Continuation 16634325
Provisional Application 62542920 · Aug 9, 2017
Related Publication 20230142099A1 · May 11, 2023
References Cited (127)
US 5914254A · Mascarenhas et al. · 1999 [cited by applicant]
US 8980267B2 · Grewal et al. · 2015 [cited by applicant]
US 9139634B2 · Morrison et al. · 2015 [cited by applicant]
US 9315567B2 · Chang et al. · 2016 [cited by applicant]
US 9492562B2 · Tavernier et al. · 2016 [cited by applicant]
US 9534056B2 · Grewal et al. · 2017 [cited by applicant]
US 9732135B2 · Tavernier et al. · 2017 [cited by applicant]
US 9878014B2 · Tavernier et al. · 2018 [cited by applicant]
US 9914759B2 · Tavernier et al. · 2018 [cited by applicant]
US 9932409B2 · Tavernier et al. · 2018 [cited by applicant]
US 10034919B2 · Tavernier et al. · 2018 [cited by applicant]
US 10035835B2 · Tavernier et al. · 2018 [cited by applicant]
US 10072059B2 · Tavernier et al. · 2018 [cited by applicant]
US 10407480B2 · Tavernier et al. · 2019 [cited by applicant]
US 10640542B2 · Tavernier et al. · 2020 [cited by applicant]
US 11566072B2 · Kley · 2023 [cited by examiner]
US 20100172868A1 · Morrison et al. · 2010 [cited by applicant]
US 20100297076A1 · Morrison et al. · 2010 [cited by applicant]
US 20110020273A1 · Chang et al. · 2011 [cited by applicant]
US 20110081341A1 · Honjo et al. · 2011 [cited by applicant]
US 20110104112A1 · Morrison et al. · 2011 [cited by applicant]
US 20110224407A1 · Langer et al. · 2011 [cited by applicant]
US 20110274658A1 · Silver et al. · 2011 [cited by applicant]
US 20130137856A1 · Steyaert et al. · 2013 [cited by applicant]
US 20130183298A1 · Le et al. · 2013 [cited by applicant]
US 20130230517A1 · Grewal et al. · 2013 [cited by applicant]
US 20140271462A1 · Ho et al. · 2014 [cited by applicant]
US 20140348789A1 · Tavernier et al. · 2014 [cited by applicant]
US 20150139951A1 · Grewal et al. · 2015 [cited by applicant]
US 20150313965A1 · Pogue et al. · 2015 [cited by applicant]
US 20180186894A1 · Tavernier et al. · 2018 [cited by applicant]
US 20180333465A1 · Tavernier et al. · 2018 [cited by applicant]
US 20180334488A1 · Tavernier et al. · 2018 [cited by applicant]
US 20180334489A1 · Tavernier et al. · 2018 [cited by applicant]
US 20190010199A1 · Tavernier et al. · 2019 [cited by applicant]
US 20190071500A1 · Kley et al. · 2019 [cited by applicant]
US 20190144553A1 · Kley et al. · 2019 [cited by applicant]
US 20190194284A1 · Kley et al. · 2019 [cited by applicant]
US 20190202934A1 · Tavernier et al. · 2019 [cited by applicant]
US 20190351021A1 · Tavernier et al. · 2019 [cited by applicant]
US 20190352406A1 · Tavernier et al. · 2019 [cited by applicant]
US 20190367575A1 · Tavernier et al. · 2019 [cited by applicant]
US 20190367604A1 · Kley et al. · 2019 [cited by applicant]
US 20200071414A1 · Kley et al. · 2020 [cited by applicant]
US 20200087411A1 · Kley et al. · 2020 [cited by applicant]
RU 2011127226A · 2013 [cited by applicant]
WO WO9102754A1 · 1991 [cited by applicant]
WO 2000055305A1 · 2000 [cited by applicant]
WO WO03033720A2 · 2003 [cited by applicant]
WO WO2006053883A1 · 2006 [cited by applicant]
WO WO2006115800A2 · 2006 [cited by applicant]
WO 2007110231A2 · 2007 [cited by applicant]
WO WO2008014612A1 · 2008 [cited by applicant]
WO WO2008124086A2 · 2008 [cited by applicant]
WO WO2009003145A1 · 2008 [cited by applicant]
WO WO2009013484A1 · 2009 [cited by applicant]
WO WO2009039409A1 · 2009 [cited by applicant]
WO WO2010036918A2 · 2010 [cited by applicant]
WO WO2010066740A1 · 2010 [cited by applicant]
WO WO2011020783A2 · 2011 [cited by applicant]
WO WO2011029870A1 · 2011 [cited by applicant]
WO WO2012170072A1 · 2012 [cited by applicant]
WO WO2013053008A2 · 2013 [cited by applicant]
WO WO2013059885A2 · 2013 [cited by applicant]
WO WO2013107791A1 · 2013 [cited by applicant]
WO WO2013134138A1 · 2013 [cited by applicant]
WO WO2014122183A1 · 2014 [cited by applicant]
WO WO2014164553A1 · 2014 [cited by applicant]
WO WO2015007520A1 · 2015 [cited by applicant]
WO WO2015007536A2 · 2015 [cited by applicant]
WO WO2015007542A1 · 2015 [cited by applicant]
WO WO2015007903A1 · 2015 [cited by applicant]
WO WO2015018528A1 · 2015 [cited by applicant]
WO WO2017077382A1 · 2017 [cited by applicant]
WO WO2017134302A2 · 2017 [cited by applicant]
WO WO2017194782A2 · 2017 [cited by applicant]
Rabia et al Understanding and overcoming trade-offs between antibody affinity, specificity, stability and solubility (Biochemical Engineering Journal 137 (2018) 365-374) (Year: 2018). [cited by examiner]
Sabir et al. Construction of naive camelids VHH repertoire in phage display-based libraryC. R. Biologies 337 (2014) 244-249 (Year: 2014). [cited by examiner]
Lloyd et al.Modelling the human immune response: performance of a 1011 human antibody repertoire against a broad panel of therapeutically relevant antigens. Protein Engineering, Design & Selection vol. 22 No. 3 pp. 159-… [cited by examiner]
Acres, et al., “Fusokine Interleukin-2/Interleukin-18, a Novel Potent Innate and Adaptive Immune Stimulator with Decreased Toxicity,” Cancer Res., vol. 65, No. 20, pp. 9536-9546, 2005. [cited by applicant]
Baba, et al., “Identification of CCR6, the Specific Receptor for a Novel Lymphocyte-Directed CC Chemokine LARC,” The Journal of Biological Chemistry, vol. 272, No. 23, pp. 14893-14898, 1997. [cited by applicant]
Barbara, et al., “Dissociation of TNF-α cytotoxic and proinflammatory activities by p55 receptor-and p75 receptor-selective TNF-α mutants,” EMBO Journal, vol. 13, No. 4, pp. 843-850, 1994. [cited by applicant]
Bork, et al., “Go hunting in sequence databases but watch out for the traps.” Trends in Genetics, vol. 12, pp. 125-427, 1996. [cited by applicant]
Bork, “Powers and Pitfalls in Sequence Analysis: The 70% Hurdle,” Genome Research, vol. 10, pp. 398-400, 2000. [cited by applicant]
Boschert, et al., “Single chain TNF derivatives with individually mutated receptor binding sites reveal differential stoichiometry of ligand receptor complex formation for TNFR1 and TNFR2,” Cellular Signalling 22 (7):10… [cited by applicant]
Bremer, et al., “Superior activity of fusion protein scFvRit:sFasL over cotreatment with rituximab and Fas agonists,” Cancer Res. 68: 597-604, 2008. [cited by applicant]
Camacho, et al., “Structure of an Interleukin-1β Mutant With Reduced Bioactivity Shows Multiple Subtle Changes in Conformation That Affect Protein-Protein Recognition,” Biochemistry, vol. 32, No. 34, pp. 8749-8757, 1993. [cited by applicant]
Coulstock, et al., “Liver-Targeting of Interferon-Alpha with Tissue Specific Domain Antibodies,” PLOS ONE, vol. 8, No. 2, pp. 1-11, 2013. [cited by applicant]
De Bruyn, et al., “Antibody-Based Fusion Proteins to Target Death Receptors in Cancer,” Cancer Letters, vol. 332, pp. 175-183, 2013. [cited by applicant]
Deffar, et al., “Nanobodies—The New Concept in Antibody Engineering,” African Journal of Biotechnology, vol. 8, No. 12, pp. 2645-2652, 2009. [cited by applicant]
Dijkmans, et al., “Murine Interferon-γ Interleukin-1 Fusion Proteins Used as Antigens for the Generation of Hybridomas Producing Monoclonal Anti-Interleukin-1 Antibodies,” Cytokine, vol. 3, No. 2, pp. 134-140, 1991. [cited by applicant]
Dimitrov, “Engineered CH2 Domains (Nanoantibodies),” mAbs, Landes Bioscience, vol. 1, No. 1, pp. 26-28, 2009. [cited by applicant]
Frey, et al., “Antibody-Based Targeting of Interferon-Alpha to the Tumor Neovasculature: A Critical Evaluation,” ntegrative Biology, vol. 3, pp. 468-478, 2011. [cited by applicant]
Garcin, et al., “High Efficiency cell-specific targeting of cytokine activity,” Nature Communications, vol. 5, No. 8, 9 pages, 2014. [cited by applicant]
Garlanda, et al., “The Interleukin-1 Family: Back to the Future,” Immunity, 39 (6): pp. 1003-1018, Dec. 12, 2013. [cited by applicant]
Holler, et al., “Two Adjacent Trimeric Fas Ligands are Required for Fas Signaling and Formation of a Death-Inducing Signaling Complex,” Molecular and Cellular Biology, vol. 23, No. 4, pp. 1428-1440, 2003. [cited by applicant]
Huang, et al., “A Trimeric Anti-HER2/neu ScFv and Tumor Necrosis Factor-[alpha] Fusion Protein Induces HER2/Neu Signaling and Facilitates Repair of Injured Epithelia,” The Journal of Pharmacology and Experimental Therap… [cited by applicant]
Idoyaga, et al., “Comparable T helper 1 (Th1) and CD8 T-cell immunity by targeting HIV gag p24 to CD8 dendritic cells within antibodies to Langerin, DEC205, and Clec9A,” PNAS, vol. 108, No. 6, pp. 2384-2389, Jan. 24, 20… [cited by applicant]
International Search Report & Written Opinion, PCT Application No. PCT/EP17/52553, dated Jul. 12, 2017, 14 pages. [cited by applicant]
Kircheis, et al., “Biological activity of mutants of human tumour necrosis factor-alpha,” Immunology, pp. 433-438, Jul. 1, 1992. [cited by applicant]
Krippner-Heidenreich, et al., “Single-Chain TNF, a TNF Derivative with Enhanced Stability and Antitumoral Activity,” The Journal of Immunology, vol. 180, pp. 8176-8183, 2008. [cited by applicant]
Lahoud, et al., “Targeting Antigen to Mouse Dendritic Cells via Clec9A Induces Potent CD4 T Cell Responses Biased toward a Follicular Helper Phenotype,” The Journal of Immunology, vol. 187, No. 2, pp. 842-850, Jul. 15, … [cited by applicant]
Loetscher, et al., “Human Tumor Necrosis Factor α (TNFα) Mutants with Exclusive Specificity for 55-kDA or 75-kDa TNF Receptors,” Journal of Biological Chemistry, American Society for Biochemistry and Molecular Biology, … [cited by applicant]
Masci, et al., “New and Modified Interferon alfas: Preclinical and Clinical Data,” Current Oncology Reports, vol. 5, pp. 108-113, 2003. [cited by applicant]
Minn, “Interferons and the Immunogenic Effects of Cancer Therapy,” Trends In Immunology, vol. 36, No. 11, pp. 725-737, Nov. 1, 2015. [cited by applicant]
Ngo, et al., “Computational Complexity, Protein Structure Prediction, and the Levinthal Paradox. The Protein Folding Problem and Tertiary Structure Prediction,” Edited by: Mertz et al., (Birkhauser, Boston), pp. 491-495… [cited by applicant]
Pan, et al., “Mutation of the IFNAR-1 Receptor Binding Site of Human IFN-α2 Generates Type I IFN Competitive Antagonists,” Biochemistry, vol. 47, pp. 12018-12027, 2008. [cited by applicant]
Patris, et al., “Nanoimmunoassay onto a screen printed electrode for HER2 breast cancer biomarker determination,” Talanta, 2014, vol. 130, pp. 164-170, 2014. [cited by applicant]
Penafuerte, et al., “The Human Ortholog of Granulocyte Macrophage Colony-Stimulating Factor and Interleukin-2 fusion Protein Induces Potent Ex Vivo Natural Killer Cell Activation and Maturation,” Cancer Res, vol. 69, No… [cited by applicant]
Picco, et al., “Targeting DNGR-1 (CLEC9A) with antibody/MUC1 peptide conjugates as a vaccine for carcinomas,” European Journal of Immunology, vol. 44, No. 7, pp. 1947-1955, Apr. 17, 2014. [cited by applicant]
Puskas, et al., “Development of an attenuated interleukin-2 fusion protein that can be activated by tumour-expressed proteases,” Immunology, vol. 133, No. 2, pp. 206-220, Jun. 23, 2011. [cited by applicant]
Rafei, et al., “A MCP1 Fusokine with CCR2-Specific Tumoricidal Activity,” Molecular Cancer, vol. 10, No. 121, pp. 1-11, 2011. [cited by applicant]
Rafei, et al., “An Engineered GM-CSF-CCL2 Fusokine Is a Potent Inhibitor of CCR2-Driven Inflammation as Demonstrated in a Murine Model of Inflammatory Arthritis,” The Journal of Immunology, vol. 183, pp. 1759-1766, 2009. [cited by applicant]
Roisman, et al., “Structure of the Interferon-Receptor Complex Determined by Distant Constraints from Double Mutant Cycles and Flexible Docking,” PNAS, vol. 98, No. 23, pp. 13231-13236, 2001. [cited by applicant]
Rovero, et al., “Insertion of the DNA for the 163-171 Peptide of IL 1 II Enables a DNA Vaccine Encoding p185 [cited by applicant]
Sancho, et al., “Identification of a dendritic cell receptor that couples sensing of necrosis to immunity,” Nature, Nature Publishing Group, United Kingdom, vol. 453, No. 7240, pp. 899-903, Apr. 16, 2009. [cited by applicant]
Schutyser, et al., “The CC Chemokine CCL20 and its Receptor CCR6,” Cytokine & Growth Factor Reviews, vol. 14, pp. 409-426, 2003. [cited by applicant]
Vaneycken, et al., “Preclinical Screening of Anti-HER2 Nanobodies for Molecular Imaging of Breast Cancer”, The ASEB Journal, vol. 25, pp. 2433-2446, 2011. [cited by applicant]
Weber, et al., “Single Amino Acid Changes that Render Human IFN-α2 Biologically Active on Mouse Cells,” The EMBO Journal, vol. 6, No. 3, pp. 591-598, 1987. [cited by applicant]
Wells, “Additivity of Mutational Effects in Proteins,” Biochemistry, vol. 29, No. 37, pp. 8509-8517, 1990. [cited by applicant]
Wesolowski, et al., “Single Domain Antibodies: Promising Experimental and Therapeutic Tools in Infection and Immunity,” Med. Microbiol. Immunol., vol. 198, pp. 157-174, 2009. [cited by applicant]
Zitvogel, et al., “Type I interferons in anticancer immunity,” The Journal of Immunology, vol. 15, No. 7, pp. 405-141, Jun. 1, 2015. [cited by applicant]
Chain B, The Catalytic Domain of Murine Urokinase-type Plasminogen Activator in Complex With the Active Site Binding Inhibitory Nanobody Nb22, PDB: 5LHR_B, downloaded from the internet, <http:www.ncbi.nlm.nih.gov/protei… [cited by applicant]
International Search Report & Written Opinion, PCT. Appl. No. PCT/US18/45741, dated Dec. 14, 2018, 15 pages. [cited by applicant]
ToIC family protein [Hydrogenovibrio crunogenus], NCBI Reference Sequence: WP_011369874.1, downloaded from the internet, <https:www.ncbi.nlm.nih.gov/protein/WP_011369874.1?reporting=genbank&log$=protalign&blast_rank=1&R… [cited by applicant]
Rashidian, et al., “Predicting the response to CTLA-4 blockade by longitudinal non-invasive monitoring of CD8 T cells”, Journal of Experimental Evidence, 214(8), pp. 2243-2255 (2017). [cited by applicant]
Rock, et al., “CDR3 Length in Antigen-specific Immune Receptors,” J. Exp. Med., vol. 179, pp. 323-328, Jan. 1994. [cited by applicant]