IP Library › Granted Patent US 12,304,961
Granted Patent B2
US 12,304,961 · App. 18/181,339 · Granted May 20, 2025

PD-L1 and OX40 binding proteins for cancer regulation

Inventors: Jhong-Jhe You (Taipei, TW); Ching-Hsuan Hsu (Taoyuan, TW); Po-Lin Huang (Taipei, TW); Hung-Tsai Kan (New Taipei, TW); Ting-Yi Chang (New Taipei, TW); Hsin-Ta Hsieh (Taipei, TW); Jeng-Horng Her (San Jose, CA)
Assignee: AP Biosciences, Inc.
C07K16/2878A61P35/00C07K16/2827A61K2039/505C07K2317/31C07K2317/52C07K2317/55C07K2317/622
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 12,304,961
App. No.
18/181,339
Granted
May 20, 2025
Kind
B2
Abstract

Provided are monospecific and bispecific proteins that bind specifically to OX40 and/or PD-L1. Exemplary proteins release the inhibition through PD-L1 and stimulate T cell through OX40. Exemplary polyvalent proteins comprise at least one OX40 binding site and at least one PD-L1 binding site. In certain embodiments, the binding sites may be linked through an immunoglobulin constant region. Anti-OX40 and anti-PD-L1 antibodies are also provided.

Claims (42)

1. An antibody or an antigen-binding portion thereof binding to OX40 (CD134), comprising:

a heavy chain variable region and a light chain variable region respectively comprising an amino acid sequence of:

amino acid 128-246 of SEQ ID NO: 10 and amino acid 1-112 of SEQ ID NO: 10.

2. The antibody or the antigen-binding portion thereof of claim 1 , wherein the antibody or the antigen-binding portion thereof is a single chain variable fragment (scFv), wherein the sequence of the scFv is SEQ ID NO: 10.

3. The antibody or the antigen-binding portion thereof of claim 1 , wherein the antibody or the antigen-binding portion thereof is a bispecific antibody.

4. The antibody or the antigen-binding portion thereof of claim 3 , wherein the bispecific antibody comprises an immune checkpoint protein binding site.

5. The antibody or the antigen-binding portion thereof of claim 4 , wherein the immune checkpoint protein binding site comprises a programmed cell death protein 1 ligand (PD-L1) binding site, PD-1 binding site, cytotoxic T-lymphocyte-associated antigen 4 (CTLA-4) binding site, or lymphocyte activation gene 3 (LAG3) binding site.

6. A bispecific antibody comprising at least one of polypeptide chain, wherein the polypeptide chain comprises:

an OX40 binding site, comprising:

a heavy chain variable region and a light chain variable region respectively comprising an amino acid sequence of amino acid 128-246 of SEQ ID NO: 10 and amino acid 1-112 of SEQ ID NO: 10;

and

a PD-L1 binding site, comprising:

a heavy chain variable region and a light chain variable region respectively comprising an amino acid sequence of SEQ ID NO: 2 and amino acid 1-111 of SEQ ID NO: 1.

7. The bispecific antibody of claim 6 , wherein the polypeptide chain further comprises:

a Fc domain;

a Fab fragment connected to a N-terminus of the Fc domain, wherein the Fab fragment comprises the PD-L1 binding site; and

a scFv connected to a C-terminus of the Fc domain, wherein the scFv comprises the OX40 binding site.

8. The bispecific antibody of claim 7 , wherein the polypeptide chain further comprises a linker between the Fc domain and the scFv.

9. The bispecific antibody of claim 6 , wherein the bispecific antibody comprises two of said polypeptide chains.

10. The bispecific antibody of claim 9 , wherein the bispecific antibody is an IgG, IgE, IgM, IgD, IgA, or IgY antibody.

11. The bispecific antibody of claim 10 , wherein the bispecific antibody is an IgG antibody.

12. The bispecific antibody of claim 11 , wherein the IgG antibody is an IgG1, IgG2, IgG3, or IgG4 antibody.

13. A pharmaceutical composition comprising the antibody or the antigen-binding portion thereof according to claim 1 , and at least one pharmaceutically acceptable carrier.

14. A pharmaceutical composition comprising the bispecific antibody according to claim 6 , and at least one pharmaceutically acceptable carrier.

15. An antibody-drug conjugate comprising:

a therapeutic agent; and

an antibody or an antigen-binding portion thereof binding PD-L1 and/or OX40, wherein the therapeutic agent is covalently conjugated to the antibody or the antigen-binding portion thereof by a linker;

wherein the antibody or the antigen-binding portion thereof is:

a) an antibody or an antigen-binding portion thereof binding to OX40, comprising:

a heavy chain variable region and a light chain variable region respectively comprising an amino acid sequence of:

amino acid 128-246 of SEQ ID NO: 10 and amino acid 1-112 of SEQ ID NO: 10;

or

b) a bispecific antibody comprising at least one polypeptide chain, wherein the polypeptide chain comprises:

an OX40 binding site, comprising:

a heavy chain variable region and a light chain variable region respectively comprising an amino acid sequence of amino acid 128-246 of SEQ ID NO: 10 and amino acid 1-112 of SEQ ID NO: 10;

and

a PD-L1 binding site, comprising:

a heavy chain variable region and a light chain variable region respectively comprising an amino acid sequence of SEQ ID NO: 2 and amino acid 1-111 of SEQ ID NO: 1.

16. A method of treating cancer comprising administering to a subject in need thereof an effective amount of the antibody or the antigen-binding portion thereof according to claim 1 .

17. The method of claim 16 , wherein the cancer is selected from the group consisting of prostate cancer, lung cancer, Non-Small Cell Lung Cancer (NSCLC), melanoma, lymphoma, breast cancer, head and neck cancer, renal cell carcinoma (RCC), and ovarian cancer.

18. A method of treating cancer comprising administering to a subject in need thereof an effective amount of the bispecific antibody according to claim 6 .

19. The method of claim 18 , wherein the cancer is selected from the group consisting of prostate cancer, lung cancer, Non-Small Cell Lung Cancer (NSCLC), melanoma, lymphoma, breast cancer, head and neck cancer, renal cell carcinoma (RCC), and ovarian cancer.

Assignments (1)
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Mar 9, 2023
From: YOU, JHONG-JHE; HSU, CHING-HSUAN; HUANG, PO-LIN; KAN, HUNG-TSAI; CHANG, TING-YI; HSIEH, HSIN-TA; HER, JENG-HORNG
To: AP BIOSCIENCES, INC.
Reel/Frame 062937/0813 →
Continuity (4)
Continuation 16871799 · May 11, 2020
Continuation PCTUS2018067868 · Dec 28, 2018
Provisional Application 62611543 · Dec 29, 2017
Related Publication 20230322939A1 · Oct 12, 2023
References Cited (113)
US 5541297A · Hansen et al. · 1996 [cited by applicant]
US 7550140B2 · Bakker et al. · 2009 [cited by applicant]
US 7943743B2 · Korman et al. · 2011 [cited by applicant]
US 7960515B2 · Min et al. · 2011 [cited by applicant]
US 9006399B2 · Liu et al. · 2015 [cited by applicant]
US 9175082B2 · Zhou et al. · 2015 [cited by applicant]
US 9475880B2 · Simons et al. · 2016 [cited by applicant]
US 9856320B2 · Cogswell et al. · 2018 [cited by applicant]
US 10058609B2 · Zhou et al. · 2018 [cited by applicant]
US 10457732B2 · Kasturirangan et al. · 2019 [cited by applicant]
US 11117972B2 · Eckelman et al. · 2021 [cited by applicant]
US 20150307617A1 · Du et al. · 2015 [cited by applicant]
US 20160166685A1 · Cheung et al. · 2016 [cited by applicant]
US 20170198051A1 · Eckelman et al. · 2017 [cited by applicant]
US 20170275362A1 · Brentjens et al. · 2017 [cited by applicant]
US 20170281765A1 · Zhou et al. · 2017 [cited by applicant]
US 20180147271A1 · Morgan et al. · 2018 [cited by applicant]
US 20180327504A1 · Al-Shamkhani et al. · 2018 [cited by applicant]
US 20180346571A1 · Gurney et al. · 2018 [cited by applicant]
US 20190112380A1 · Chaudhary · 2019 [cited by applicant]
US 20200299389A1 · Her et al. · 2020 [cited by applicant]
CN 104080809A · 2014 [cited by applicant]
CN 104470949A2 · 2015 [cited by applicant]
CN 104736168A · 2015 [cited by applicant]
CN 106103486A · 2016 [cited by applicant]
CN 106999583A · 2017 [cited by applicant]
CN 107074953A · 2017 [cited by applicant]
JP 2008544755A · 2008 [cited by applicant]
JP 2011505836A · 2011 [cited by applicant]
JP 2013538057A · 2013 [cited by applicant]
JP 2014527814A · 2014 [cited by applicant]
JP 2015519375A · 2015 [cited by applicant]
JP 2017514461A · 2017 [cited by applicant]
WO 03106498A2 · 2003 [cited by applicant]
WO 2013038191A2 · 2013 [cited by applicant]
WO 2013181634A2 · 2013 [cited by applicant]
WO 2016090337A1 · 2016 [cited by applicant]
WO 2016187216A1 · 2016 [cited by applicant]
WO 2017068181A1 · 2017 [cited by applicant]
WO 2017087547A1 · 2017 [cited by applicant]
WO 2017096120A1 · 2017 [cited by applicant]
WO 2017123673A2 · 2017 [cited by applicant]
WO 2017172981A2 · 2017 [cited by applicant]
WO 2017193032A2 · 2017 [cited by applicant]
WO 2019168947A1 · 2019 [cited by applicant]
WO 2019184909A1 · 2019 [cited by applicant]
Reeck, “Homology' in Proteins and Nucleic Acids: A Terminology Muddle and A Way out of it” (Letter to the Editor), Cell, 1987, vol. 50, Cell Press. [cited by applicant]
Rudikoff et al., “Single amino acid substitution altering antigen-binding specificity,” 1982, Proc Natl Acad Sci. USA, vol. 79, pp. 1979-1983. [cited by applicant]
Tamura et al., “Structural Correlates of an Anticarcinoma Antibody: Identification of Specificity-Determining Residues (SDRs) and Development of a Minimally Immunogenic Antibody Variant by Retention of SDRs Only,” 2000,… [cited by applicant]
Yarilin, “The Principles of Immunology”, Moscow: Meditsina, 1999, pp. 169-174, vol. 608. [cited by applicant]
Dondelinger et al., “Understanding the Significance and Implications of Antibody Numbering and Antigen-Binding Surface/Residue Definition”, Frontiers in Immunology, 2018, vol. 9, doi.org/10.3389/fimmu.2018.02278. [cited by applicant]
Tingsong, “Advances in Treatment of Gastric Cancer”, Tongji University Press, May 31, 2017, p. 144. [cited by applicant]
Zhang et al., “Fc Engineering Approaches to Enhance the Agonism and Effector Functions of an Anti-OX40 Antibody”, Journal of Biological Chemistry, pp. 27134-27146, vol. 291(53). [cited by applicant]
Weinberg et al., “Anti-OX40 (CD134) administration to nonhuman primates: immunostimulatory effects and toxicokinetic study”, Journal of Immunotherapy, Nov.-Dec. 2006, vol. 29(6), pp. 575-585. [cited by applicant]
Guo et al., “PD-1 Blockade and OX40 Triggering Synergistically Protects against Tumor Growth in a Murine Model of Ovarian Cancer”, PLOS ONE, Feb. 27, 2014, vol. 9(2). [cited by applicant]
Granier et al., “Mechanisms of action and rationale for the use of checkpoint inhibitors in cancer”, ESMO Open, 2017. [cited by applicant]
Shrimali et al., “Concurrent PD-1 Blockade Negates the Effects of OX40 Agonist Antibody in Combination Immunotherapy through Inducing T-cell Apoptosis”, Cancer Immunology Research, 2017, pp. 755-766. [cited by applicant]
Hui et al., “Research on an anti-human OX40 eliciting monoclonal antibody and preliminary study on its biological ability”, Modern Immunology, May 31, 2010, pp. 195-201, vol. 30(3). [cited by applicant]
Wei et al., “Comprehensive therapeutic strategies regarding immune checkpoint”, Chin J Clin Oncol, Aug. 15, 2017, pp. 782-786, vol. 44(15). [cited by applicant]
Ping et al., “Research progress of new target OX40 in tumor immunotherapy”, Chinese Journal of Microbiology and Immunology, Mar. 31, 2017, pp. 240-244, vol. 37(3). [cited by applicant]
Geng et al., “Advances in the application of bispecific antibody drugs”, Progress in Biotechnology, Nov. 25, 2015, pp. 420-424, vol. 5(6). [cited by applicant]
Nagaya et al., “Near infrared photoimmunotherapy with avelumab, an anti-programmed death-ligand 1 (PD-L1) antibody”, Oncotarget, 2017, pp. 8807-8817, vol. 8, No. 5. [cited by applicant]
Bargou et al., “Tumor Regression in Cancer Patients by Very Low Doses of a T Cell-Engaging Antibody”, Science, 2008, pp. 974-977, vol. 321, No. 5891. [cited by applicant]
Bretscher et al., “A Theory of Self-Nonself Discrimination”, Science, 1970, pp. 1042-1049, vol. 169, No. 3950. [cited by applicant]
Bretscher, “A two-step, two-signal model for the primary activation of precursor helper T cells”, Proceedings of the National Academy of Sciences of the United States of America, 1999, pp. 185-190, vol. 96, No. 1. [cited by applicant]
Chames et al., “Bispecific antibodies for cancer therapy: The light at the end of the tunnel?”, mAbs, 2009, pp. 539-547, vol. 1, No. 6. [cited by applicant]
Croft, “Costimulation of T cells by OX40, 4-1BB, and CD27”, Cytokine & Growth Factor Reviews, 2003, pp. 265-273. vol. 14, Nos. 3-4. [cited by applicant]
Demarest et al., “Antibody therapeutics, antibody engineering, and the merits of protein stability”, Current Opinion in Drug Discovery & Development, 2008, pp. 675-687, vol. 11, No. 5. [cited by applicant]
Fridman et al., “The immune contexture in human tumours: impact on clinical outcome”, Nature Reviews, Cancer, 2012, pp. 298-306, vol. 12, No. 4. [cited by applicant]
Goldenberg et al., “Cancer imaging and therapy with bispecific antibody pretargeting”, Update on Cancer Therapeutics, 2007, pp. 19-31, vol. 2, No. 1. [cited by applicant]
Heiss et al., “The trifunctional antibody catumaxomab for the treatment of malignant ascites due to epithelial cancer: results of a prospective randomized phase II/III trial”, International Journal of Cancer, 2010, pp. … [cited by applicant]
Hollander, “Bispecific antibodies for cancer therapy”, Immunotherapy, 2009, pp. 211-222, vol. 1, No. 2. [cited by applicant]
Jenkins et al., “Antigen Presentation by Chemically Modified Splenocytes Induces Antigen-Specific T Cell Unresponsiveness in Vitro and in Vivo”, Journal of Experimental Medicine, 1987, pp. 302-319, vol. 165, No. 2. [cited by applicant]
Karpovsky et al., “Production of target-specific effector cells using hetero-cross-linked aggregates containing anti-target cell and anti-Fc gamma receptor antibodies”, Journal of Experimental Medicine, 1984, pp. 1686-1… [cited by applicant]
King et al., “A new Hu-PBL model for the study of human islet alloreactivity based on NOD-scid mice bearing a targeted mutation in the IL-2 receptor gamma chain gene”, Clinical Immunology, 2008, pp. 303-314, vol. 126, N… [cited by applicant]
Lafferty et al., “A New Analysis of Allogeneic Interactions”, The Australian Journal of Experimental Biology and Medical Science, 1975, pp. 27-42, vol. 53, Pt. 1. [cited by applicant]
Larkin et al., “Combined Nivolumab and Ipilimumab or Monotherapy in Untreated Melanoma”, The New England Journal of Medicine, 2015, pp. 23-34, vol. 373, No. 1. [cited by applicant]
Lenschow et al., “CD28/B7 System of T Cell Costimulation”, Annual Review of Immunology, 1996, pp. 233-258, vol. 14. [cited by applicant]
Liang et al., “PD-L1 and PD-L2 have distinct roles in regulating host immunity to cutaneous leishmaniasis”, European Journal of Immunology, 2006, pp. 58-64, vol. 36, No. 1. [cited by applicant]
Linch et al., “OX40 agonists and combination immunotherapy: putting the pedal to the metal”, Frontiers in Oncology, 2015, 14 pages, vol. 5, Article No. 34. [cited by applicant]
Lu et al., “A Fully Human Recombinant IgG-like Bispecific Antibody to Both the Epidermal Growth Factor Receptor and the Insulin-like Growth Factor Receptor for Enhanced Antitumor Activity”, The Journal of Biological Che… [cited by applicant]
Müller et al., “Improved Pharmacokinetics of Recombinant Bispecific Antibody Molecules by Fusion to Human Serum Albumin”, The Journal of Biological Chemistry, 2007, pp. 12650-12660, vol. 282, No. 17. [cited by applicant]
Pardoll, “The blockade of immune checkpoints in cancer immunotherapy”, Nature Review, Cancer, 2012, pp. 252-264, vol. 12, No. 4. [cited by applicant]
Paterson et al., “Antigens of Activated Rat T Lymphocytes Including a Molecule of 50,000 Mr Detected Only on CD4 Positive T Blasts”, Molecular Immunology, 1987, pp. 1281-1290, vol. 24, No. 12. [cited by applicant]
Perez et al., “Specific targeting of cytotoxic T cells by anti-T3 linked to anti-target cell antibody”, Nature, 1985, pp. 354-356, vol. 316, No. 6026. [cited by applicant]
Ridgway et al., “Knobs-into-holes' engineering of antibody CH3 domains for heavy chain heterodimerization”, Protein Engineering, 1996, pp. 617-621, vol. 9, No. 7. [cited by applicant]
Staerz et al., “Hybrid antibodies can target sites for attack by T cells”, Nature, 1985, pp. 628-631, vol. 314, No. 6012. [cited by applicant]
Sznol et al., “Antagonist Antibodies to PD-1 and B7-H1 (PD-L1) in the Treatment of Advanced Human Cancer”, Clinical Cancer Research, 2013, pp. 1021-1034, vol. 19, No. 5. [cited by applicant]
Thakur et al., “Cancer therapy with bispecific antibodies: Clinical experience”, Current Opinion in Molecular Therapeutics, 2010, pp. 340-349, vol. 12, No. 3. [cited by applicant]
Topalian et al., “Survival, Durable Tumor Remission, and Long-Term Safety in Patients With Advanced Melanoma Receiving Nivolumab”, Journal of Clinical Oncology, 2014, pp. 1020-1030, vol. 32, No. 10. [cited by applicant]
Watts, “TNF/TNFR Family Members in Costimulation of T Cell Responses”, Annual Review of Immunology, 2005, pp. 23-68, vol. 23. [cited by applicant]
Weinberg et al., “Target Organ-Specific Up-Regulation of the Mrc OX-40 Marker and Selective Production of Th1 Lymphokine mRNA by Encephalitogenic T Helper Cells Isolated from the Spinal Cord of Rats with Experimental Au… [cited by applicant]
“Atezolizumab”, Wikipedia, pp. 1-5, retrieved from https://en.wikipedia.org/wiki/Atezolizumab on May 4, 2021. [cited by applicant]
Azevedo et al., “First-line atezolizumab monotherapy in patients with advanced BRAFV600 wild-type melanoma”, Pigment Cell & Melanoma Research, 2021. [Abstract]. [cited by applicant]
Bahleda et al., “Long-Term Safety and Clinical Outcomes of Atezolizumab in Head and Neck Cancer: Phase la Trial Results”, Annals of Oncology, Sep. 2017, pp. 373, vol. 28:5. [Abstract]. [cited by applicant]
Geoerger et al., “Atezolizumab for children and young adults with previously treated solid tumours, non-Hodgkin lymphoma, and Hodgkin lymphoma (iMATRIX): a multicentre phase 1-2 study”, The Lancet Oncology, 2019, pp. 13… [cited by applicant]
“Highlights of Prescribing Information: Tecentriq (atezolizumab) injection, for intravenous use”, U.S. Food and Drug Administration, 2016, pp. 1-38. [cited by applicant]
Lafon et al., “Atezolizumab for the treatment of renal cell carcinoma”, Expert Opinion on Biological Therapy, 2020, pp. 679-686, vol. 20:7. [cited by applicant]
Petrylak et al., “Safety and Clinical Activity of Atezolizumab in Patients with Metastatic Castration-Resistant Prostate Cancer: A Phase I Study”, Clinical Cancer Research, 2021, pp. 3360-3369, vol. 27:12. [cited by applicant]
Rico et al., “Atezolizumab for the treatment of colorectal cancer: the latest evidence and clinical potential”, Expert Opinion on Biological Therapy, 2018, pp. 449-457, vol. 18:4. [cited by applicant]
Bulliard et al., “OX40 engagement depletes intratumoral Tregs via activating FcyRs, leading to antitumor efficacy”, Immunology and Cell Biology, 2014, pp. 475-480, vol. 92. [cited by applicant]
Lee et al. “Molecular mechanism of PD-1/PD-L1 blockade via anti-PD-L1 antibodies atezolizumab and durvalumab”, Scientific Reports, pp. 1-12, vol. 7. [cited by applicant]
Alves Costa Silva et al., “New pathways in immune stimulation: targeting OX40,” ESMO Open, 2020, pp. 1-8, vol. 5, article No. e000573. [cited by applicant]
Bell et al., “OX40 signaling in head and neck squamous cell carcinoma: Overcoming immunosuppression in the tumor microenvironment,” Oral Oncology, 2016, pp. 1-10, vol. 52. [cited by applicant]
Ramser et al., “High OX40 expression in recurrent ovarian carcinoma is indicative for response to repeated chemotherapy,” BMC Cancer, 2018, pp. 1-9, vol. 18, article No. 425. [cited by applicant]
Yokouchi et al., “Anti-OX40 monoclonal antibody therapy in combination with radiotherapy results in therapeutic antitumor immunity to murine lung cancer,” Cancer Sci., 2008, pp. 361-367, vol. 99:2. [cited by applicant]
Chen et al., “Enhancement and destruction of antibody function by somatic mutation: unequal occurrence is controlled by V gene combinatorial associations”, The EMBO Journal, 1995, pp. 2784-2794, vol. 14, No. 12, Oxford … [cited by applicant]
Kranz et al., “Restricted reassociation of heavy and light chains from hapten-specific monoclonal antibodies”, Proc. Natl. Acad. Sci. USA, 1981, pp. 5807-5811, vol. 78, No. 9. [cited by applicant]
Amminmaki et al., “Crystal Structure of a Recombinant Anti-estradiol Fab Fragment in Complex with 17(beta)-Estradiol”, The Journal of Biological Chemistry, 2001, pp. 36687-36694, vol. 276, No. 39, The American Society f… [cited by applicant]
MacCallum et al., “Antibody-antigen Interactions: Contact Analysis and Binding Site Topography”, J. Mol. Biol., 1996, pp. 732-745, Academic Press Limited. [cited by applicant]
Nezlin, “Biochmistry of Antibodies”, 1970, p. 160, Plenum Press: New York. [cited by applicant]
Pearson, “An Introduction to Sequence Similarity (‘Homology’) Searching”, Current Protocols in Bioinformatics, 2013, pp. 1-8, John Wiley & Sons, Inc. [cited by applicant]
Tan et al., “Distinct PD-L1 binding characteristics of therapeutic monoclonal antibody durvalumab”, Protein & Cell, vol. 9, Issue 1, Jan. 2018, pp. 135-139. [cited by applicant]