EP-4739775-A1 - RECOMBINANT GLYCOSIDASES, PHARMACEUTICAL COMPOSITIONS, AND USES IN MANAGING AUTOIMMUNE OR INFLAMMATORY CONDITIONS

EP4739775A1EP 4739775 A1EP4739775 A1EP 4739775A1EP-4739775-A1

Abstract

This disclosure relates to recombinant immunoglobulin endoglycosidases and uses in managing diseases or conditions associated with immune dysregulation or inflammation. In certain embodiments, the recombinant immunoglobulin endoglycosidases comprise sequences disclosed herein. In certain embodiments, this disclosure relates to pharmaceutical compositions and methods of treating or preventing an autoimmune disease or inflammatory condition comprising administering to a subject in need thereof an effective amount of a recombinant immunoglobulin endoglycosidase, or nucleic acids or vector encoding the same, as disclosed herein.

Inventors

SUNDBERG, ERIC
SASTRE, Diego
DU, Jonathan
BOURNAZOS, Stylianos
RAVETCH, JEFFREY V.

Assignees

Emory University
The Rockefeller University

Dates

Publication Date: 20260513
Application Date: 20240708

Claims (1)

CLAIMS What is claimed is: 1. A recombinant immunoglobulin endoglycosidase comprising peptide motifs of the following amino acid sequences from an N-terminal to C-terminal order sequentially, YYRTWRDK (SEQ ID NO: 1), TDIP, (SEQ ID NO: 2), LDGLDX’DME (SEQ ID (SEQ ID NO: 4), NRFL (SEQ ID NO: 5), and DRDG (SEQ ID NO: 6), wherein any amino acid and provided the immunoglobulin endoglycosidase does not contain an amino acid sequence KESV (SEQ ID NO: 48, contained in CP40) between LDGLDX’DME (SEQ ID NO: 3) and SCQF (SEQ ID NO: 4). 2. A recombinant immunoglobulin endoglycosidase of claim 1 comprising peptide motifs of the following amino acid sequences from an N-terminal to C-terminal order sequentially, AXXXXXPLXXXXGXXXXXG (SEQ ID NO: 7), PIXXXYYRTWRDKXIXXXXXD (SEQ ID NO: 8), TDIPXXXXXXSLXHVXD, (SEQ ID NO: 9), QXSDXXFWXTFXXXYXPXLXXRGTXVXXTXXXXXXL (SEQ ID NO: 10) YXXXAXXXXXXYVXXHXLDGLDXDME, (SEQ ID NO: 11) LRXXMXAXXXLXGPXXXXN (SEQ ID NO: 12) LXYXTXXNAQXXXXXXV (SEQ ID NO: 13), AXXVXYVXXQTY (SEQ ID NO: 14) WNXXRXXXXSCQFXXXYAXPEEXDX (SEQ ID NO: 15), NRFLXAXGXGXVXXXXAXXXAXWXP (SEQ ID NO: 16), and GXKGGXXXYAXDRDGXTYDXXDXXTLXXTXFXXXKR (SEQ ID NO: 17), wherein X is individually and independently at each position is any amino acid. 3. The recombinant immunoglobulin endoglycosidase of claim 1 comprising the amino acid sequence CU43) AALSNAPLAASPGQADKVGAQATCAAKPIFFGYYRTWRDKAIELNDGDKWKDKLHTK LTDIPEQVDMVSLFHVPDNQKSDQRFWETFDKEYHPTLKERGTKVVRTIGAKLLLNKIK EKGLYGQ SREDD SK YREI AHEVYEEYVAKHNLDGLD VDMELREVEKYTNLRWQLRKI MGAFSELMGPKAPGNAGKKPGDDGYKYLIYDTFDNAQLAQVALVADVVDYVLAQTY DKGTEESITRVWNGFRDKINSCQFLAGYAHPEENDTNRFLTAIGDVDTSGAMNVAAWK PEGGEKGGTFAYALDRDGRTYDGDDLTTLKPTDFAFTKRAIELTKGISLTDLG (SEQ ID NO: 19). 4. A recombinant immunoglobulin endoglycosidase of claim 1 comprising the amino acid sequence of (CP258) ADLSQAPLKASPGHADKVGVQTTCDAKPIFFGYYRTWRDKAIQLKDDDPWKDKLQVK LTDIPEHVNMVSLFHVEDNQKSDDQFWETFRKEYQPKLKERGTRVVRTVGAQLLLNKI KEKGLYGRSVEDDYKYREIARDIYKKYVTDHNLDGLDVDMELRKVEKRIDLQWQLRKI MGAFSELMGPKAPANEGKKPGHEGKKPGHEGYKYLIYDTFDNAQTSQVGLVADLVDY VLAQTYDKGTKESIDQVWNGFRDKINSCQFMAGYAHPEENDTNRFLTAVGEVNKSGA MQVAEWKPDNGVKGGTFAYALDRDGRTYDGDDFTTLKPTDFAFTKRAIELTTGESSTD LG (SEQ ID NO: 20). 5. A recombinant immunoglobulin endoglycosidase of claim 1 comprising the amino acid sequence of (CM49) ASPTSLPLPEHKGFHPDAGERSDCEPIVTAYYRTWRDKEIQQLPDDRVGPNVIAMTDIPH NIDVLSLIHVPDHQQSDQQFWATFASTYLPELHRRGTKVLYTLDISAVLDPSLKPTSSPEE YAAHAQRLVDKYVRPHKLDGLDIDMERDLSFDQRIVLRNTMRALTQLVGPFSNTNTLL TYNTNRNAQKAYIDEVAQYVNYVFVQTYSVTDPNEIQYSYWNTYRFYLSSCQFLPSYA NPEEFDRNRFLGAIGPVEETAAVKIAGWQPFQGGVKGGIMTYAIDRDGMTYDQPDISTL RVTTFPVIKRVTAVLKAKKFAAAK (SEQ ID NO: 21). 6. A recombinant immunoglobulin endoglycosidase comprising peptide motifs of the following amino acid sequences from an N-terminal to C-terminal order sequentially, PVFF (SEQ ID NO: 22), YYRTWRDKAI (SEQ ID NO: 23), LTDIP (SEQ ID NO: 24), MVSLFHA (SEQ ID NO: 25), VRTT (SEQ ID NO: 26), HNLDGLDVDME, (SEQ ID NO: 27), GPKA (SEQ ID NO: 28), LIYDTFD (SEQ ID NO: 29), VDYVL (SEQ ID NO: 30), SCQF (SEQ ID NO: 4), GYAHPEE (SEQ ID NO: 31), NRFETAIG (SEQ ID NO: 32), DRDGRTY(SEQ ID NO: 33), TFKR (SEQ ID NO: 34). 7. A recombinant immunoglobulin endoglycosidase of claim 6 comprising peptide motifs of the following amino acid sequences from an N-terminal to C-terminal order sequentially, PXXXXXSLXXXXGXXXXXG (SEQ ID NO: 35), PVFFXYYRTWRDKAIXXXXXD (SEQ ID NO: 36), LTDIPXXVXMVSLFHAXN, (SEQ ID NO: 37), YXSDXXFWXTFXXXYXPXLKXRGTXVRTTXXAXXLL (SEQ ID NO: 38) DXXYREXAXXXXXXYVXXHNLDGLDVDME, (SEQ ID NO: 39) WXIRKXMXAXSELXGPKAXXN (SEQ ID NO: 40) GYKXLIYDTFDXXXXXQ (SEQ ID NO: 41), AXXVDYVLXQTY (SEQ ID NO: 42) WNXXRXXXXSCQFXXGYAHPEEXDT (SEQ ID NO: 43), NRFETAIGXGXVXXSXAMXVAXWXP (SEQ ID NO: 44), and GGXKGGXFXYAXDRDGRTYXXDDXXXXKXTDFXTFKR (SEQ ID NO: 45), wherein X is individually and independently at each position is any amino acid. 8. A recombinant immunoglobulin endoglycosidase of claim 6 comprising the amino acid sequence of (CR39) PAIAETSLRGGTGQHETPGSQKCDSEPVFFAYYRTWRDKAIVQNESDKPNRANKIALTDI PQHVNMVSLFHAGNDHYQSDAEFWKTFDDVYYPELKRRGTRVVRTISATELLKETDSL RAVGVDTDAADYREVAEKIKKEYVDAHNLDGLDVDMEVLHLERNWRSSWEKRWRIR KTMAALSELLGPKADVNQGKKKDDSGYKFLIYDTFDDVERSQIRAIAELVDYVLPQTYK SGKAEIDQLWNQSKGILSSCQFVPGYAHPEEGDTVNRFETAIGDVDSSKAMEVAAWQP AGGEKGGAFVYAIDRDGRTYGEDDLKNVKETDFSFTKRGAALARSVTFAKAK (SEQ ID NO: 46). 9. A fusion protein comprising any of the immunoglobulin endoglycosidases as provided in claims 1-8. 10. A nucleic acid encoding a recombinant immunoglobulin endoglycosidase as provided in claims 1-8 in operable combination with a heterologous promoter. 11. The nucleic acid of claim 10 which is mRNA, RNA, or DNA. 12. A live attenuated Corynebacterium pseudotuberculosis strain comprising a recombinant immunoglobulin endoglycosidase as provided in any of claims 1-9. 13. A vector comprising a nucleic acid of claim 10 or encoding a recombinant immunoglobulin endoglycosidase as provided in claims 1-9. 15. A method of treating or preventing a disease or condition associates mediated by IgG antibodies or with altered Asn297 IgG glycosylation comprising administering to a subject in need thereof an effective amount of a recombinant immunoglobulin endoglycosidase as provided in claims 1-9, or nucleic acid, vector, or attenuated Corynebacterium pseudotuberculosis strain encoding a recombinant immunoglobulin endoglycosidase as provided in claims 10-12. 16. The method of claim 15 wherein the disease or condition an autoimmune diseases and chronic inflammatory condition such as, including rheumatoid arthritis, systemic lupus erythematosus, inflammatory bowel disease, autoimmune liver disease, juvenile idiopathic arthritis, osteoarthritis, spondyloarthropathies, neonatal lupus, lupus nephritis, Sjogren’s syndrome, antineutrophil cytoplasmic antibody (ANCA)-associated antineutrophil cytoplasmic antibody (ANCA)-associated vasculitis (AAV), Crohn’s disease, ulcerative colitis, Hashimoto’s thyroiditis, multiple sclerosis, Guillain-Barre syndrome, chronic inflammatory demyelinating polyneuropathy, Myasthenia gravis, Lambert-Eaton myasthenic syndrome, autoimmune hemolytic anemia, and antiphospholipid syndrome. 17. The method of claim 15 wherein the disease or conditions is associated with increased or decreased galactosylation, sialylation, fucosylation, and/or the presence of bisecting N- acetylglucosamine (GlcNAc) chains. 18. A method of treating or preventing a viral infection comprising administering to a subject in need thereof an effective amount of a recombinant immunoglobulin endoglycosidase as provided in claims 1 -9, or nucleic acid, vector, or attenuated Corynebacterium pseudotuberculosis strain encoding a recombinant immunoglobulin endoglycosidase as provided in claims 10-12. 19. A method of treating or preventing antibody-mediated depletion of B cells comprising administering to a subject in need thereof an effective amount of a recombinant immunoglobulin endoglycosidase as provided in claims 1-9, or nucleic acid, vector, or attenuated Corynebacterium pseudotuberculosis strain encoding a recombinant immunoglobulin endoglycosidase as provided in claims 10-12. 20. A method of treating or preventing antibody-mediated cytotoxicity against CD4 T cells comprising administering to a subject in need thereof an effective amount of a recombinant immunoglobulin endoglycosidase as provided in claims 1-9, or nucleic acid, vector, or attenuated Corynebacterium pseudotuberculosis strain encoding a recombinant immunoglobulin endoglycosidase as provided in claims 10-12.

Description

RECOMBINANT GLYCOSIDASES, PHARMACEUTICAL COMPOSITIONS, AND USES IN MANAGING AUTOIMMUNE OR INFLAMMATORY CONDITIONS CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit of U.S. Provisional Application No. 63/525,476 filed July 7, 2023 and U.S. Provisional Application No. 63/584,527 filed September 22, 2023. The entirety of each of these applications is hereby incorporated by reference for all purposes. STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT This invention was made with government support under Al 149297 awarded by the National Institutes of Health. The government has certain rights in the invention. INCORPORATION-BY-REFERENCE OF MATERIAL SUBMITTED AS AN XML FILE VIA THE OFFICE ELECTRONIC FILING SYSTEM The Sequence Listing associated with this application is provided in XML format and is hereby incorporated by reference into the specification. The name of the XML file containing the Sequence Listing is 23002PCT.xml. The XML file is 48 KB, was created on July 5, 2024, and is being submitted electronically via the USPTO patent electronic filing system. BACKGROUND Humans produce antibodies containing immunoglobulin G (IgG) proteins which are abundantly present in the blood. As part of the immune system, IgG antibodies play a central role in protecting the body from pathogens by recognizing the presence of biological material that is not a “self-molecule.” When IgG antibodies specifically bind a pathogen, a series of biological events occur that cause the body to attack the pathogen. However, environmental factors, genetic defects, and aging sometimes inadvertently directs these same events to occur abnormally. Thus, the presence of undesirable IgG antibodies are often the cause of autoimmune diseases and inflammatory disorders. Recombinant therapeutic antibody therapies exist for certain autoimmune diseases and inflammatory disorders; however, treatments are not always universally effective and sometimes have side effects. Thus, there is a need to identify improvements. Human TgG proteins contain variable regions and constant regions (Fc). The Fc region contains an asparagine amino acid at position 297, (Asn297 or N297), which is commonly substituted to contain different chains of sugar units, also referred to as a glycan. The state of glycosylation at Asn297 is reported to be involved regulating immunological events. See Flevaris et al. Int J Mol Sci, 2022, 23, 5180. Flevaris et al. report immunoglobulin GN-glycan biomarkers for autoimmune diseases. Int J Mol Sci, 2022, 23, 5180 Shadnezhad et al. report CP40 from Corynebacterium pseudotuberculosis is an endo- -N- acetylglucosaminidase. BMC Microbiology, 2016, 16:261. Yamin et al. report human FcyRIIIa activation on splenic macrophages drives dengue pathogenesis in mice. Nat Microbiol, 2023, 8(8): 1468-1479. References cited herein are not an admission of prior art. SUMMARY This disclosure relates to immunoglobulin endoglycosidases and uses in managing diseases or conditions associated with immune dysregulation. In certain embodiments, this disclosure relates to methods of cleaving or altering glycans attached to immunoglobulins comprising contacting a glycosylated immunoglobulin and an immunoglobulin endoglycosidase disclosed herein providing a glycan cleaved immunoglobulin. In certain embodiments, this disclosure relates to recombinant immunoglobulin endoglycosidases comprising sequences disclosed herein. In certain embodiments, this disclosure relates to nucleic acids and vectors encoding immunoglobulin endoglycosidases disclosed herein. In certain embodiments, this disclosure relates to pharmaceutical compositions comprising recombinant immunoglobulin endoglycosidases disclosed herein and/or nucleic acids or vector encoding the same. In certain embodiments, this disclosure relates to methods of treating or preventing an autoimmune disease or condition associated with glycosylated immunoglobulins comprising administering to a subject in need thereof an effective amount of a recombinant immunoglobulin endoglycosidase, or nucleic acids or vectors encoding the same, as disclosed herein. In certain embodiments, the recombinant immunoglobulin endoglycosidase comprises the amino acid sequence of (CU43) AALSNAPLAASPGQADKVGAQATCAAKPIFFGYYRTWRDKAIELNDGDKWKDK LHTKLTDIPEQVDMVSLFHVPDNQKSDQRFWETFDKEYHPTLKERGTKVVRTIGAKLLL NKIKEKGL YGQ SREDD SK YREI AHE V YEE Y V AK HNLDGLD VDMELRE VEK YTNLR WQ LRKIMGAFSELMGPKAPGNAGKKPGDDGYKYLIYDTFDNAQLAQVALVADVVDYVLA QTYDKGTEESITRVWNGFRDKINSCQFLAGYAHPEENDTNRFLTAIGDVDTSGAMNVA AWKPEGGEKGGTFAYALDRDGRTYDGDDLTTLKPTDFAFTKRAIELTKGISLTDLG (SEQ ID NO: 19). In certain embodiments, the recombinant immunoglobulin endoglycosidase comprises the amino acid sequence of (CP258) ADLSQAPLKASPGHADKVGVQTTCDAKPIFFGYYRTWRDKAIQLKDDDPWKDKLQVK LTDIPEHVNMVSLFHVEDNQKSDDQFWETFRKEYQPKLKERGTRVVRTVGAQLLLNKI KEKGLYGRSVEDDYKYREIARDIYKKYVTDHNLDGLDVDMELRKVEKRIDLQWQLRKI MGAFSELMGPKAPANEGKKPGHEGKKPGHEGYKYLIYDTFDNAQTSQVGLVADL