Granted Patent
B2
US 12,351,636 · App. 17/010,178 · Granted Jul 8, 2025
Fusion immunomodulatory proteins and methods for making same
Inventors:
Nagaraj Govindappa (Karnataka, IN); Maria Melina Soares (Karnataka, IN); Kedarnath Sastry (Karnataka, IN)
Assignee:
Biocon Ltd.
C07K16/2863C07K14/705C07K14/71C07K16/2818C12N15/62A61K38/00C07K2317/40C07K2317/526C07K2317/90C07K2319/00
View Patent ↗
Loading inventors, assignments & file history…
Abstract
The present invention relates generally to the field of generating recombinant chimeric fusion proteins to be used in the cancer therapy, and more specifically, to fusion molecules of Anti-EGFR1-TGFβRII, Anti-EGFR1-PD1 and Anti-CTLA4-PD1 and methods of generating same, wherein the methods reduce production costs and increase homogeneity of the recombinant chimeric fusion proteins.
Claims (2)
1. A fusion protein comprising a targeting moiety and an immunomodulating moiety exhibiting bi-specificity, wherein the targeting moiety and the immunomodulating moiety are linked by an amino acid spacer encoded by nucleotide sequence SEQ ID NO: 4, wherein the immunomodulating moiety is encoded by nucleotide sequence SEQ ID NO: 3 or SEQ ID NO: 7, wherein the targeting moiety is an anti-EGFR1 antibody comprising a heavy chain and a light chain encoded by nucleotide sequences consisting of SEQ ID NO: 1 and SEQ ID NO: 2, respectively, and wherein the immunomodulating moiety is attached via the amino acid spacer to the C-terminus of the heavy chain or the light chain.
2. A fusion protein comprising a targeting moiety and an immunomodulating moiety exhibiting bi-specificity, wherein the targeting moiety and the immunomodulating moiety are linked by an amino acid spacer encoded by nucleotide sequence SEQ ID NO: 4, wherein the immunomodulating moiety is encoded by nucleotide sequence SEQ ID NO: 3 or SEQ ID NO: 7, wherein the targeting moiety is an anti-CTLA-4 antibody comprising a heavy chain and a light chain encoded by nucleotide sequences consisting of SEQ ID NO: 5 and SEQ ID NO: 6, respectively, and wherein the immunomodulating moiety is attached via the amino acid spacer to the C-terminus of the heavy chain or the light chain.
Assignments (1)
ASSIGNMENT OF ASSIGNOR'S INTEREST
Recorded Jan 29, 2024
From: GOVINDAPPA, NAGARAJ; SOARES, MARIA MELINAS; SASTRY, KEDARNATH
To: BIOCON LIMITED
Reel/Frame 066283/0577 →
Continuity (4)
Division
14771308
Provisional Application
61777016
· Mar 12, 2013
References Cited (118)
US 7141651B2
· Gillies et al.
· 2006
[cited by applicant]
US 8815247B2
· Govindappa et al.
· 2014
[cited by applicant]
US 9340617B2
· Govindappa et al.
· 2016
[cited by applicant]
US 9452149B2
· Nilsson et al.
· 2016
[cited by applicant]
US 9758582B2
· Govindappa et al.
· 2017
[cited by applicant]
US 9809651B2
· Govindappa et al.
· 2017
[cited by applicant]
US 9988456B2
· Govindappa et al.
· 2018
[cited by applicant]
US 20100297697A1
· Ambrosius et al.
· 2010
[cited by applicant]
CN 101511373A
· 2009
[cited by applicant]
EP 0361902
· 1989
[cited by applicant]
JP H02163096
· 1990
[cited by applicant]
RU 2120475C1
· 1998
[cited by applicant]
WO WO199833914
· 1998
[cited by applicant]
WO WO0155163A1
· 2001
[cited by applicant]
WO WO0158957A2
· 2001
[cited by applicant]
WO WO2002083854
· 2002
[cited by applicant]
WO WO2008077889
· 2008
[cited by applicant]
WO WO2009027471
· 2009
[cited by applicant]
WO WO2009039409
· 2009
[cited by applicant]
WO WO2009114110
· 2009
[cited by applicant]
WO WO2011102845
· 2011
[cited by applicant]
WO WO2011109789
· 2011
[cited by applicant]
WO WO2012033953A1
· 2012
[cited by applicant]
WO WO2012147048A2
· 2012
[cited by applicant]
WO WO2012147053
· 2012
[cited by applicant]
WO WO2013004841
· 2013
[cited by applicant]
WO WO201316469
· 2013
[cited by applicant]
WO WO2014164427A1
· 2014
[cited by applicant]
Altschul, Stephen et al. “Basic Local Alignment Search Tool.” 1990,
[cited by applicant]
Birch, J. R. et al. “Antibody production.” Advanced Drug Delivery Reviews, vol. 58, No. 5-6, Aug. 7, 2006.
[cited by applicant]
Bitter, Grant et al. “Expression and Secretion Vectors for Yeast.” 1987,
[cited by applicant]
Carton et al. “Codon engineering for improved antibody expression in mammalian cells.” Protein Expression and Purification, Academic Press, vol. 55, No. 2, Sep. 8, 2007.
[cited by applicant]
Casi, Giulio et al. “Antibody drug conjugates: Basic concepts, examples and future perspectives.” Journal of Controlled Release, vol. 161, No. 2, Jan. 10, 2012.
[cited by applicant]
Dick, Lawrence W. et al. “C-Terminal Lysine Variants in Fully Human Monoclonal Antibodies: Investigation of Test Methods and Possible Causes.” Biotechnology and Bioengineering, vol. 100, No. 6, Aug. 15, 2008.
[cited by applicant]
Dong, Mei et al. “Role of transforming growth factor-ß in hematologic malignancies.” 2006,
[cited by applicant]
Fath, Stephan, et al. “Multiparameter RNA and Codon Optimization: A Standardized Tool to Access and Enhance Autologous Mammalian Gene Expression.” 2011,
[cited by applicant]
Flies, Dallas B. et al. “Blockade of the B7-H1/PD-1 Pathway for Cancer Immunotherapy.” 2011,
[cited by applicant]
Kalwy, S. et al. “Toward more efficient protein expression.” Molecular Biotechnology, Humana Press, Inc., vol. 34, No. 2, Sp. Iss. SI, Oct. 1, 2006.
[cited by applicant]
Kotsopoulou, E. et al. “Optimised mammalian expression through the coupling of codon adaptation with gene amplification; maximum yields with minimum effort.” Journal of Biotechnology, vol. 146, No. 4, Apr. 15, 2010.
[cited by applicant]
Lee, Robert J. et al. “Delivery of liposomes into cultured KB cells via folate receptor-mediated endocytosis.” 1994,
[cited by applicant]
Lee, Robert J. et al. “Folate-mediated tumor cell targeting of liposome-entrapped doxorubicin in vitro.” 1995
[cited by applicant]
Luo, Jun et al. “Probing of C-Terminal Lysine Variation in a Recombinant Monoclonal Antibody Production Using Chinese Hamster Ovary Cells with Chemically Defined Media.” Biotechnology and Bioengineering, vol. 109, No. 9…
[cited by applicant]
Mackett, Michael et al. “Vaccinia virus: a selectable eukaryotic cloning and expression vector.” 1982,
[cited by applicant]
Mackett, Michael et al. “General method for production and selection of infectious vaccinia virus recombinants expressing foreign genes.” 1984,
[cited by applicant]
Nomi, Takeo et al. “Clinical significance and therapeutic potential of the programmed death-1 ligand/programmed death-1 pathway in human pancreatic cancer.” 2007,
[cited by applicant]
Ortiz-Sanchez, Elizabeth et al. “Antibody cytokine fusion proteins: applications in cancer therapy.” Expert Opinion on Biological Therapy, vol. 8, No. 5, May 1, 2008.
[cited by applicant]
Okazaki, Taku et al. “PD-1 and PD-1 ligands: from discovery to clinical Application.” 2007,
[cited by applicant]
Panicali, Dennis et al. “Construction of poxviruses as cloning vectors: insertion of the thymidine kinase gene from herpes simplex virus into the DNA of infectious vaccine virus.” 1982,
[cited by applicant]
Pardoll, Drew M. “The blockage of immune checkpoints in cancer immunotherapy.” 2012,
[cited by applicant]
Raab, David et al. “The GeneOptimizer Algorithm: using a sliding window approach to cope with the vast sequence space in multiparameter DNA sequence optimization.” 2010,
[cited by applicant]
Sundaram, Shanmuuga et al. “An innovative approach for the characterization of the isoforms of a monoclonal antibody product.” MABS, vol. 3, No. 6, Nov. 1, 2011.
[cited by applicant]
Thompson, R. Houston, et al. “Costimulatory B7-H1 in renal cell carcinoma patients: Indicator of tumor aggressiveness and potential therapeutic target.” 2004,
[cited by applicant]
Wrzesinski, Stephen et al. “Transforming Growth Factor-ß and the Immune Response: Implications for Anticancer Therapy.” 2007,
[cited by applicant]
Altschul, et al. Basic Local Alignment Search Tool. J. Mol. Biol., 1990, pp. 403-410.
[cited by applicant]
Carton et al., Codon engineering for improved antibody expression in mammalian cells, Protein Expression and Purification, Sep. 2007, pp. 279-286.
[cited by applicant]
Cocco, et al. hl-con1, a factor VII-IgGFc chimeric protein targeting tissue factor for immunotherapy of uterine serous papillary carcinoma. British journal of cancer, 2010, pp. 812-819.
[cited by applicant]
European Extended Search Report, corresponding to European Divisional Patent Application No. 18212095, dated Mar. 8, 2019, 11 pages.
[cited by applicant]
European Office Action, corresponding to European Divisional Patent Application No. 18153517.0, dated Mar. 22, 2018, 10 pages.
[cited by applicant]
Fath, et al. Multiparameter RNA and Codon Optimization: A Standardized Tool to Access and Enhance Autologous Mammalian Gene Expression, 2011, PloSone, pp. 1-14.
[cited by applicant]
Giulio, et al. Antibody-drug conjugates: Basic concepts, examples and future perspectives. Journal of Controlled Release, 2012, pp. 422-428.
[cited by applicant]
International Preliminary Report on Patentability for Application No. PCT/US2014/022404 dated Sep. 15, 2015, 11 pages.
[cited by applicant]
International Search Report and Written Opinion for Application No. PCT/US2014/022404 dated Aug. 1, 2014, 15 pages.
[cited by applicant]
Japanese Office Action, corresponding to Japanese Patent Application No. 2016-500960, dated Oct. 18, 2017, 14 pages.
[cited by applicant]
Liu, et al. Heterogeneity of monoclonal antibodies. Pharmaceutical Sciences, American Pharmaceutical Assoc., 2007, pp. 2426-2447.
[cited by applicant]
Luo, J. et al. Comparative metabolite analysis to understand lactate metabolism shift in Chinese hamster ovary cell culture process. Biotechnology and Bioengineering, 2011, pp. 146-156.
[cited by applicant]
Office Action, issued by the European Patent Office for corresponding European Divisional Patent Application No. 18153517.0, dated Mar. 22, 2018, 10 pages.
[cited by applicant]
Office Action, issued by the European Patent Office for corresponding European Patent Application No. 1373158.2, dated Jun. 6, 2017, 9 pages.
[cited by applicant]
Office Action, issued by the Russian Patent Office for corresponding Russian Patent Application No. 2014145158, dated Feb. 14, 2017, 6 pages.
[cited by applicant]
Qian, et al. Cell Culture and Gene Transcription Effects of Copper Sulfate on Chinese Hamster Ovary Cells. Biotechnology Progress, 2011, pp. 1190-1194.
[cited by applicant]
Russian Office Action, corresponding to Russian Divisional Patent Application No. 2018117558, issued by the Patent Office of the Russian Federation dated Jan. 29, 2019, 5 pages.
[cited by applicant]
Author Unknown, Cathepsin L, filed with Notice of Opposition in EP2844667 (Application No. 13734158.2) on Jul. 11, 2019, 3 pages, downloaded/retrieval dated May 23, 2019 from https://www.signmaaldrich.com/life-science/b…
[cited by applicant]
Author Unknown, Opposition against EP2844667 (Application No. 13734158.2) further efficacy data provided by patentee, filed with Notice of Opposition in EP2844667 on Jan. 27, 2020, 5 pages, downloaded/retrieval date unk…
[cited by applicant]
Author Unknown, Overview of the embodiments shown in the Figures of D1 filed with Notice of Opposition in EP2844667 (Application No. 13734158.2) on Jul. 11, 2019, 2 pages, downloaded/retrieval date unknown.
[cited by applicant]
Author Unknown, Sequence alignment of i) the amino acid sequence translated from SEQ ID No. 16 and II) the fused amino acid sequence consisting of SEQ ID No. 5 and SEQ ID No. 3 (in this order), annotated copy of D18, da…
[cited by applicant]
Author Unknown, Sequence alignment of i) the amino acid sequence translated from SEQ ID No. 16 and ii) the fused amino acid sequence consisting of SEQ ID No. 5 and SEQ ID No. 3 (in this order), dated Feb. 3, 2020, 2 pag…
[cited by applicant]
Author Unknown, Sequence alignment of SEQ ID No. 104 of D1 v. SEQ ID No. 3 of the patent in suit, 1 page downloaded/retrieval dated Feb. 28, 2019 from https://www.ebi.ac.uk/Tools/services/rest/emboss stretcher/result/em…
[cited by applicant]
Author Unknown, Sequence alignment of SEQ ID No. 2 of D1 v. SEQ ID No. 3 of the patent in suit, 2 pages downloaded/retrieval dated Feb. 28, 2019 from https://www.ebi.ac.uk/Tools/services/rest/emboss stretcher/result/emb…
[cited by applicant]
Author Unknown, Sequence alignment of SEQ ID No. 2 of D1 v. SEQ ID No. 4 of the patent in suit, 2 pages downloaded/retrieval dated Feb. 28, 2019 from https://www.ebi.ac.uk/Tools/services/rest/emboss stretcher/result/emb…
[cited by applicant]
Author Unknown, Sequence alignment of SEQ ID No. 2 of D1 v. SEQ ID No. 5 of the patent in suit, 2 pages downloaded/retrieval dated Feb. 28, 2019 from https://www.ebi.ac.uk/Tools/services/resUemboss stretcher/resulUembos…
[cited by applicant]
Author Unknown, Sequence alignment of SEQ ID No. 4 of the patent in suit and SEQ ID No. 87 of D1, 1 page downloaded/retrieval dated Feb. 28, 2019 from https://www.ebi.ac.uk/Tools/services/rest/emboss stretcher/result/em…
[cited by applicant]
Author Unknown, Sequence alignment of SEQ ID No. 71 of D1 v. SEQ ID No. 6 of the patent in suit, 1 page downloaded/retrieval dated Feb. 28, 2019 from https://www.ebi.ac.uk/Tools/services/rest/emboss stretcher/result/emb…
[cited by applicant]
Author Unknown, stability data provided by patentee, filed with Notice of Opposition in EP2844667 on Jan. 27, 2020, 7 pages, downloaded/retrieval date unknown.
[cited by applicant]
Author Unknown, Synopsis of the independent claims of EP2844667B1 (Application No. 13734158.2) and U.S. Pat. No. 8,815,247 B2, 1 page, downloaded/retrieval date unknown.
[cited by applicant]
Author Unknown, Technical Report No. DDL/TR/BR.14.09015/19/003, Report title: To evaluate role of FmAb2 in immune cell activation, using an invitro tumour-immune cell coculture assay system, filed with Notice of Opposit…
[cited by applicant]
Author Unknown, Technical Report No. IPL/TR/BR.14.09015/19/001, Report title: Evaluation of anti-tumor activity of FmAb2 in HT-29 (V600E) tumor xenograft model, filed with Notice of Opposition in EP2844667 (Application …
[cited by applicant]
Cederholm-Williams, S.A., “Concentration of plasminogen and antiplasmin in plasma and serum,” J Clin Pathol 1981; 34:979-981.
[cited by applicant]
Decision Denying Inter Partes Review for Case IPR2016-00175 Patent 8,815,247 B2, filed with Notice of Opposition in EP2844667 (Application No. 13734158.2) on Jul. 11, 2019, 11 pages.
[cited by applicant]
Decision of the Opposition & Instruction dated Mar. 12, 2021 for EP Application No. 13734158.2 (EP2844667), 1 page.
[cited by applicant]
Decision Rejecting the Opposition dated Mar. 12, 2021 for EP Application No. 13734158.2 (EP2844667), 2 pages.
[cited by applicant]
EPO Communication dated Mar. 11, 2019, enclosing Notice of Opposition for EP13734158.2 (EP2844667), 42 pages.
[cited by applicant]
Grounds for the Decision (Annex) Opposition for EP Application No. 13734158.2 (EP2844667) dated Mar. 12, 2021, 16 pages.
[cited by applicant]
Hsieh, et al., Micro-heterogeneity of the monoclonal antibody Erbitux ((TM)) characterized by ion exchange chromatography, isoelectric focusing, peptide mapping, and LC-MS/MS, Abstracts of Papers of The American Chemica…
[cited by applicant]
Keil, Specificity of Proteolysis, Springer-Verlag Berlin Heidelberg 1992, pp. 6, 12, 67, 108 (8 total pages).
[cited by applicant]
Keil, Specificity of Proteolysis, Springer-Verlag Berlin Heidelberg 1992, pp. 6, 12, 67, 108 (8 total pages), annotated copy.
[cited by applicant]
Magdelaine-Beuzelin, et al., “Structure-function relationships of the variable domains of monoclonal antibodies approved for cancer treatment,” Crit Rev Oncol Hematol. Dec. 2007; 64(3):210-25. Epub Jul. 12, 2007.
[cited by applicant]
Nakano, et al., “Molecular genetics of vaccinia virus: demonstration of marker rescue,” Proc Natl Acad Sci USA., vol. 79(5), pp. 1593-1596, Mar. 1982.
[cited by applicant]
Notice of Opposition for EP13734158.2 (EP2844667), dated and filed Feb. 28, 2019, with Exhibits, 709 total pages.
[cited by applicant]
Office Action, issued by the European Patent Office for EP Application No. 13734158.2, dated Jul. 27, 2016, 7 pages.
[cited by applicant]
Podar, et al., “Inhibition of the TGF-beta signaling pathway in tumor cells,” Recent Results Cancer Res. 2007; 172:77-97.
[cited by applicant]
Schrama, David, et al., “Antibody targeted drugs as cancer therapeutics,” Nature Reviews Drug Discovery (2006); 5.2: 147-159.
[cited by applicant]
Summons to attend oral proceedings for EP Application No. 13734158.2 (EP2844667), dated Jun. 22, 2020, 13 pages.
[cited by applicant]
Summons to attend oral proceedings for EP Application No. 13734158.2 (EP2844667), dated Jun. 6, 2017, 9 pages.
[cited by applicant]
Termination of Opposition Proceedings for EP13734158.2 (EP2844667) dated Mar. 30, 2023, 1 page.
[cited by applicant]
Written submission of ABG Patentes dated Nov. 13, 2017 for EP13734158.2 (EP2844667), 11 pages.
[cited by applicant]
Yao, et al., “Contribution of B7-H1/PD-1 Co-inhibitory pathway to T-Cell dysfunction in Cancer,” Tumor Induced Immune Suppression: Mechanisms and Therapeutic Reversal 2008 (pp. 29-40). Springer New York.
[cited by applicant]
Author Unknown, Sequence alignment of i) the amino acid sequence translated from SEQ ID No. 16 and ii) the fused amino acid sequence consisting of SEQ ID No. 5 and SEQ ID No. 3 (in this order), annotated copy of D18, da…
[cited by applicant]
Author Unknown, Sequence alignment of i) the amino acid sequence translated from SEQ ID No. 16 and ii) the fused amino acid sequence consisting of SEQ ID No. 5 and SEQ ID No. 3 (in this order), dated Feb. 3, 2020, 2 pag…
[cited by applicant]