[go: up one dir, main page]

WO2025002299A1 - Compositions et procédés pour inhiber l'expression du facteur b du complément (cfb) - Google Patents

Compositions et procédés pour inhiber l'expression du facteur b du complément (cfb) Download PDF

Info

Publication number
WO2025002299A1
WO2025002299A1 PCT/CN2024/102170 CN2024102170W WO2025002299A1 WO 2025002299 A1 WO2025002299 A1 WO 2025002299A1 CN 2024102170 W CN2024102170 W CN 2024102170W WO 2025002299 A1 WO2025002299 A1 WO 2025002299A1
Authority
WO
WIPO (PCT)
Prior art keywords
cfb
subject
dsrna
agent
dsrna agent
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
PCT/CN2024/102170
Other languages
English (en)
Inventor
Dongxu Shu
Pengcheng Patrick Shao
Shiwei Xia
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Shanghai Argo Biopharmaceutical Co Ltd
Original Assignee
Shanghai Argo Biopharmaceutical Co Ltd
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Shanghai Argo Biopharmaceutical Co Ltd filed Critical Shanghai Argo Biopharmaceutical Co Ltd
Publication of WO2025002299A1 publication Critical patent/WO2025002299A1/fr
Anticipated expiration legal-status Critical
Pending legal-status Critical Current

Links

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P13/00—Drugs for disorders of the urinary system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/7088—Compounds having three or more nucleosides or nucleotides
    • A61K31/713—Double-stranded nucleic acids or oligonucleotides
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
    • A61K45/06—Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/54—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic compound
    • A61K47/549—Sugars, nucleosides, nucleotides or nucleic acids
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
    • C12N15/113—Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2310/00—Structure or type of the nucleic acid
    • C12N2310/10—Type of nucleic acid
    • C12N2310/14—Type of nucleic acid interfering nucleic acids [NA]
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2310/00—Structure or type of the nucleic acid
    • C12N2310/30—Chemical structure
    • C12N2310/31—Chemical structure of the backbone
    • C12N2310/315—Phosphorothioates
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2310/00—Structure or type of the nucleic acid
    • C12N2310/30—Chemical structure
    • C12N2310/32—Chemical structure of the sugar
    • C12N2310/321—2'-O-R Modification
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2310/00—Structure or type of the nucleic acid
    • C12N2310/30—Chemical structure
    • C12N2310/32—Chemical structure of the sugar
    • C12N2310/322—2'-R Modification
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2310/00—Structure or type of the nucleic acid
    • C12N2310/30—Chemical structure
    • C12N2310/35—Nature of the modification
    • C12N2310/351—Conjugate
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2750/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA ssDNA viruses
    • C12N2750/00011—Details
    • C12N2750/14011—Parvoviridae
    • C12N2750/14111—Dependovirus, e.g. adenoassociated viruses
    • C12N2750/14141—Use of virus, viral particle or viral elements as a vector
    • C12N2750/14143—Use of virus, viral particle or viral elements as a vector viral genome or elements thereof as genetic vector

Definitions

  • the invention relates, in part, to compositions and methods that can be used to inhibit a complement factor B (CFB) gene expression.
  • CFB complement factor B
  • Complement was first discovered in the 1890s when it was found to aid or “complement” the killing of bacteria by heat-stable antibodies present in normal serum.
  • the complement system or pathway is part of the innate immune system of host defence against invading pathogens. It mainly consists of more than 30 proteins that are either present as soluble proteins in the blood or are present as membrane-associated proteins.
  • complement activation pathways Three main pathways of complement activation have been recognized and are referred to as classical pathway, alternative pathway and lectin pathway. Activation of complement leads to a sequential cascade of enzymatic reactions, known as complement activation pathways resulting in the formation of the potent anaphylatoxins C3a and C5a that elicit a plethora of physiological responses that range from chemoattraction to apoptosis.
  • complement activation pathways resulting in the formation of the potent anaphylatoxins C3a and C5a that elicit a plethora of physiological responses that range from chemoattraction to apoptosis.
  • complement was thought to play a major role in innate immunity where a robust and rapid response is mounted against invading pathogens.
  • complement also plays an important role in adaptive immunity involving T and B cells that help in elimination of pathogens, in maintaining immunologic memory preventing pathogenic reinvasion, and is involved in numerous human pathological states.
  • complement activation occurs inherently at a low level (spontaneous cleavage of C3 to yield C3a and C3b) and is reinforced in the presence of microbes via an enzymatic cascade converting inactive forms of enzymes (zymogenes) into their active counterparts.
  • C3 convertases is a complex of C3b and complement factor B (CFB, Factor B) . Once formed, a C3 convertase can convert large amounts of C3 into its cleavage products C3a and C3b within short amount of time.
  • C3 convertase which is a complex of C3b and Factor B has originally been described in the context of the alternative pathway, but may form also in the context of the other two pathways.
  • Factor B is also a constituent of C5 convertase, acomplex which converts C5, a more downstream component of the pathway, into its active form.
  • Complement Factor B also known as CFB or “factor B”
  • CFB Complement Factor B
  • Binding of CFB to C3b renders CFB susceptible to cleavage by Factor D, forming the serine protease C3Bb, which is itself a C3 convertase, leading to an amplification loop for C3 activation.
  • CFB is primarily synthesised in the liver, as well as in low levels at several extrahepatic sites.
  • C3 glomerulopathy systemic lupus erythematosus (SLE) , Lupus Nephritis, IgA nephropathy, diabetic nephropathy, polycystic kidney disease, membranous nephropathy, age-related macular degeneration, atypical hemolytic uremic syndrome, thrombotic microangiopathy, myasthenia gravis, ischemia and reperfusion injury, paroxysmal nocturnal hemoglobinuria, rheumatoid arthritis, immune complex-mediated glomerulonephritis (IC-mediated GN) , post-infectious glomerulonephritis (PIGN) , ischemia/reperfusion injury , antineutrophil cytoplasmic autoantibodies-associated vasculitis (ANCA-AV) , dysbiotic periodontal disease, malaria
  • Eculizumab has been shown to be effective for the treatment of paroxysmal nocturnal hemoglobinuria (PNH) , atypical hemolytic uremic syndrome (aHUS) , and Myasthenia Gravis, and is currently being evaluated in clinical trials for additional complement component-associated diseases, eculizumab therapy requires weekly high dose infusions followed by biweekly maintenance infusions at a high cost. There is thus a high unmet need for medical treatments of complement mediated or associated diseases.
  • C3 is a pivotal factor in the complement pathway activation.
  • CFB Inhibiting expression of factors such as CFB which are involved in C3 activation therefore presents a promising therapeutic strategy for many complement-mediated diseases.
  • CFB RNAi agents disclosed herein for for treating diseases, disorders, and conditions associated with complement activation by, for example, activation of complement factor B activity.
  • the present disclosure features novel CFB gene-specific RNAi agents, compositions that include CFB RNAi agents, and methods for inhibiting expression of a CFB gene in vitro and/or in vivo using the CFB RNAi agents and compositions that include CFB RNAi agents described herein.
  • the CFB RNAi agents described herein can selectively and efficiently decrease, inhibit, or silence expression of a CFB gene in a subject, e.g., a human or animal subject.
  • a double-stranded ribonucleic acid (dsRNA) agent for inhibiting expression of CFB wherein the dsRNA agent including a sense strand and an antisense strand, wherein the sense strand comprises at least 15 contiguous nucleotides differing by no more than 1, 2 or 3 nucleotides from the nucleotide sequence of SEQ ID NO: l, 3 or 5 and the antisense strand comprises at least 15 contiguous nucleotides differing by no more than 1, 2 or 3 nucleotides from the nucleotide sequence of SEQ ID NO: 2, 4 or 6, wherein the sense strand and the antisense strand can be partially, substantially, or fully complementary to each other.
  • the sense strand and the antisense strand can be partially, substantially, or fully complementary to each other.
  • the dsRNA agent includes a sense strand and an antisense strand forming a double stranded region, wherein said antisense strand comprises a region of complementarity to a CFB RNA transcript which comprises at least 15 contiguous nucleotides differing by no more than 1, 2, or 3 nucleotides from any one of the antisense sequences listed in any one of Tables 1-3.
  • the dsRNA agent includes a sense strand and an antisense strand forming a double stranded region, wherein said antisense strand comprises a region of complementarity to a CFB RNA transcript which comprises at least 15 contiguous nucleotides from any one of the antisense sequences listed in any one of Tables 1-3.
  • the dsRNA agent including a sense strand and an antisense strand, nucleotide positions 2 to 18 in the antisense strand including a region of complementarity to a CFB RNA transcript, wherein the region of complementarity includes at least 15, 16, 17, 18, 19, 20 or 21 contiguous nucleotides that differ by 0, 1, 2, or 3 nucleotides from one of the antisense sequences listed in one of Tables 1-3, and optionally including a targeting ligand.
  • a double-stranded ribonucleic acid (dsRNA) agent for inhibiting expression of CFB wherein the dsRNA agent includes a sense strand and an antisense strand, wherein the sense strand comprises at least 15, 16, 17, 18, 19, 20 or 21 contiguous nucleotides that differ by 0, 1, 2, or 3 nucleotides from any one of the nucleotide sequences of nucleotides 483-513, 486-516, 491-521, 483-521, 513-543, 987-1017, 989-1019, 1317-1347, 2237-2267, 2439-2469 of SEQ ID NO: 1, and the antisense strand comprises at least 15, 16, 17, 18, 19, 20 or 21 contiguous nucleotides differing by 0, 1, 2, or 3 nucleotides from the corresponding nucleotide sequence of SEQ ID NO: 2, wherein the sense strand and the antisense strand can be partially, substantially, or fully complementary to each other.
  • the sense strand and the antisense strand can
  • a double-stranded ribonucleic acid (dsRNA) agent for inhibiting expression of CFB wherein the sense strand comprises at least 15, 16, 17, 18, 19, 20 or 21 contiguous nucleotides that differ by 0, 1, 2, or 3 nucleotides from any one of the nucleotide sequences of nucleotides 488-508, 491-511, 496-516, 488-516, 518-538, 992-1012, 994-1014, 1322-1342, 2242-2262, 2444-2464 of SEQ ID NO: 1, and the antisense strand comprises at least 15, 16, 17, 18, 19, 20 or 21 contiguous nucleotides differing by 0, 1, 2, or 3 nucleotides from the corresponding nucleotide sequence of SEQ ID NO: 2.
  • dsRNA double-stranded ribonucleic acid
  • a double-stranded ribonucleic acid (dsRNA) agent for inhibiting expression of CFB wherein the sense strand comprises at least 15, 16, 17, 18, or 19 contiguous nucleotides that differ by 0, 1, 2, or 3 nucleotides from any one of the nucleotide sequences of nucleotides 490-508, 493-511, 498-516, 490-516, 520-538, 994-1012, 996-1014, 1324-1342, 2244-2262, 2446-2464 of SEQ ID NO: 1, and the antisense strand comprises at least 15, 16, 17, 18, or 19 contiguous nucleotides differing by 0, 1, 2, or 3 nucleotides from the corresponding nucleotide sequence of SEQ ID NO: 2.
  • dsRNA double-stranded ribonucleic acid
  • a double-stranded ribonucleic acid (dsRNA) agent for inhibiting expression of CFB wherein the sense strand comprises at least 15, 16, 17, 18, or 19 contiguous nucleotides that differ by 0, 1, 2, or 3 nucleotides from any one of the nucleotide sequences of nucleotides 489-507, 492-510, 497-515, 489-515, 519-537, 993-1011, 995-1013, 1323-1341, 2243-2261, 2445-2463 of SEQ ID NO: 1, and the antisense strand comprises at least 15, 16, 17, 18, or 19 contiguous nucleotides differing by 0, 1, 2, or 3 nucleotides from the corresponding nucleotide sequence of SEQ ID NO: 2.
  • dsRNA double-stranded ribonucleic acid
  • the CFB RNA transcript is SEQ ID NO: 1.
  • the antisense strand of the dsRNA agent is at least substantially complementary to any one of a target region of SEQ ID NO: 1 and is provided in any one of Tables 1-3. In some embodiments, the antisense strand of the dsRNA agent is fully complementary to any one of a target region of SEQ ID NO: 1 and is provided in any one of Tables 1-3. In some embodiments, the dsRNA agent includes a sense strand sequence set forth in any one of Tables 1-3, wherein the sense strand sequence is at least substantially complementary to the antisense strand sequence in the dsRNA agent.
  • the dsRNA agent includes a sense strand sequence set forth in any one of Tables 1-3, wherein the sense strand sequence is fully complementary to the antisense strand sequence in the dsRNA agent.
  • the dsRNA agent includes an antisense strand sequence set forth in any one of Tables 1-3.
  • the dsRNA agent includes the sequences set forth as a duplex sequence in any of Tables 1-3.
  • the dsRNA agent wherein the sense strand and the antisense strand can be partially, substantially, or fully complementary to each other.
  • the dsRNA agent includes at least one modified nucleotide.
  • all or substantially all of the nucleotides of the antisense strand are modified nucleotides.
  • at least one of the modified nucleotides comprises: 2’-O-methyl nucleotide, 2’-fluoro nucleotide, 2’-deoxy nucleotide, 2’ 3’-seco nucleotide mimic, locked nucleotide, unlocked nucleic acid nucleotide (UNA) , glycol nucleic acid nucleotide (GNA) , 2’-F-Arabino nucleotide, 2’-methoyxyethyl nucleotide, abasic nucleotide, ribitol, inverted nucleotide, inverted abasic nucleotide, inverted 2’-Ome nucleotide, inverted 2’-deoxy nucleot
  • the dsRNA agent includes an E-vinylphosphonate nucleotide at the 5′end of the guide strand.
  • the dsRNA agent includes at least one phosphorothioate internucleoside linkage.
  • the sense strand includes at least one phosphorothioate internucleoside linkage.
  • the antisense strand includes at least one phosphorothioate internucleoside linkage.
  • the sense strand includes 1, 2, 3, 4, 5, or 6 phosphorothioate internucleoside linkages.
  • the antisense strand includes 1, 2, 3, 4, 5, or 6 phosphorothioate internucleoside linkages.
  • the 5' end of antisense strand includes 2 phosphorothioate internucleoside linkages.
  • the 3' end of antisense strand includes 2 phosphorothioate internucleoside linkages. In some embodiments, the 5' end of antisense strand and 3' end of antisense strand includes independently 2 phosphorothioate internucleoside linkages.
  • nucleotides of the sense strand and the antisense strand are modified nucleotides.
  • the antisense strand comprises 15 or more modified nucleotides independently selected from a 2’-O-methyl nucleotide, a 2’-fluoro nucleotide and an UNA modified nucleotide, wherein less than 6 modified nucleotides are 2’-fluoro nucleotides.
  • the antisense strand comprises 3 or 5 2’-fluoro nucleotides, preferably, the antisense strand comprises 5 2’-fluoro nucleotides.
  • the sense strand comprises 15 or more modified nucleotides independently selected from a 2’-O-methyl nucleotide and a 2’-fluoro nucleotide, wherein less than 4 modified nucleotides are 2’-fluoro nucleotides. In certain embodiments, the sense strand comprises 3 2’-fluoro nucleotides.
  • the antisense strand comprises 15 or more modified nucleotides independently selected from a 2’-O-methyl nucleotide and a 2’-fluoro nucleotide, wherein at least 14 modified nucleotides are 2’-O-methyl nucleotides and the nucleotides at positions 2, 5, 7, 11, 12, 14, 16 and/or 18 counting from the first matching position of the 5’ end of the antisense strand are independently a 2’-fluoro nucleotide.
  • the antisense strand comprises at least one UNA modified nucleotide and 5 2’-fluoro nucleotides.
  • the antisense strand comprises one UNA modified nucleotide at position 7 and 5 2’-fluoro nucleotides at positions 2, 5, 12, 14 and 16 counting from the first matching position of the 5’ end, and the rest 2’-O-methyl nucleotides. In some embodiments, the antisense strand comprises one UNA modified nucleotide at position 7 and 5 2’-fluoro nucleotides at positions 2, 5, 12, 14 and 18 counting from the first matching position of the 5’ end, and the rest 2’-O-methyl nucleotides.
  • the antisense strand comprises one UNA modified nucleotide at position 7 and 5 2’-fluoro nucleotides at positions 2, 5, 11, 14 and 16 counting from the first matching position of the 5’ end, and the rest 2’-O-methyl nucleotides. In some embodiments, the antisense strand comprises 5 2’-fluoro nucleotides at positions 2, 7, 12, 14 and 16 counting from the first matching position of the 5’ end, and the rest 2’-O-methyl nucleotides. In some embodiments, the antisense strand comprises 5 2’-fluoro nucleotides at positions 2, 7, 11, 14 and 16 counting from the first matching position of the 5’ end, and the rest 2’-O-methyl nucleotides.
  • the antisense strand comprises 5 2’-fluoro nucleotides at positions 2, 5, 12, 14 and 16 counting from the first matching position of the 5’ end, and the rest 2’-O-methyl nucleotides. In some embodiments, the antisense strand comprises 5 2’-fluoro nucleotides at positions 2, 5, 12, 14 and 18 counting from the first matching position of the 5’ end, and the rest 2’-O-methyl nucleotides.
  • the sense strand comprises 15 or more modified nucleotides independently selected from a 2’-O-methyl nucleotide and a 2’-fluoro nucleotide, preferably, wherein at least 18 modified nucleotides are 2’-O-methyl nucleotides and the nucleotides at positions 9, 11 and/or 13 counting from the first matching position of the 3’end of the sense strand are 2’-fluoro nucleotides.
  • the sense strand comprises at least 18 modified nucleotides are 2’-O-methyl nucleotides and the nucleotides at positions 8, 11 and/or 13 counting from the first matching position of the 3’ end of the sense strand are 2’-fluoro nucleotides.
  • the modified sense strand is a modified sense strand sequence set forth in one of Tables 2-3.
  • the modified antisense strand is a modified antisense strand sequence set forth in one of Tables 2-3.
  • the dsRNA agent includes at least one modified nucleotide and further includes one or more targeting groups or linking groups.
  • the one or more targeting groups or linking groups are conjugated to the sense strand.
  • the targeting group or linking group includes N-acetyl-galactosamine (GalNAc) .
  • the targeting group has a structure:
  • n are independently selected from 1 or 2.
  • the targeting group has a structure:
  • the dsRNA agent includes a targeting group that is conjugated to the 5’-terminal end of the sense strand. In some embodiments, the dsRNA agent includes a targeting group that is conjugated to the 3'-terminal end of the sense strand.
  • the antisense strand includes one inverted abasic residue at 3’-terminal end.
  • the sense strand includes one or two inverted abasic residues and/or one or two imann residues at 3’ or/and 5’ terminal end.
  • each end of the sense strand includes one inverted abasic residue.
  • each end of the sense strand includes one imann residue.
  • at the 5' terminal end of the sense strand includes one inverted abasic residue or imann residues, wherein inverted abasic residue or imann residues is linked to an adjacent nucleotide via a phosphorothioate linkage to the 5' terminal end of the nucleotide sequence of the sense strand.
  • the sense strand further includes targeting group linked to an inverted abasic residue or an imann residue at the 5' terminal end of the sense strand, wherein targeting group is linked to an adjacent inverted abasic residue or imann via a phosphorothioate linkage, and optionally targeting group is N-acetyl-galactosamine (GalNAc) .
  • targeting group is linked to an adjacent inverted abasic residue or imann via a phosphorothioate linkage
  • optionally targeting group is N-acetyl-galactosamine (GalNAc) .
  • at the 5' terminal end of the sense strand includes one inverted abasic residue, wherein inverted abasic residue is linked to an adjacent nucleotide via a phosphorothioate linkage to the 5' terminal end of the nucleotide sequence of the sense strand.
  • the sense strand further includes targeting group linked to an inverted abasic residue at the 5' terminal end of the sense strand, wherein targeting group is linked to an adjacent inverted abasic residue via a phosphorothioate linkage, and optionally targeting group is N-acetyl-galactosamine (GalNAc) , each strand is independently 21 nucleotides in length.
  • targeting group is linked to an adjacent inverted abasic residue via a phosphorothioate linkage
  • optionally targeting group is N-acetyl-galactosamine (GalNAc)
  • each strand is independently 21 nucleotides in length.
  • the dsRNA agent has two blunt ends.
  • at least one strand includes a 3’ overhang of at least 1 nucleotide. In some embodiments, at least one strand includes a 3’ overhang of at least 2 nucleotides.
  • the dsRNA comprises a duplex selected from AV02373, AV02375, AV02379, AV02388, AV02411, AV02464, AV02554, AV02584, AV06327, AV06328, AV06329, and wherein duplex optionally including a targeting ligand.
  • the dsRNA comprises a duplex selected from the group consisting of AD01093, AD01093-1, AD01093-2, AD01094, AD01096, AD01393, AD01393-1, AD01396, AD01399, AD01412, AD01420, AD01420-1.
  • a double-stranded ribonucleic acid (dsRNA) agent for inhibiting expression of CFB wherein the dsRNA agent including a sense strand and an antisense strand, wherein the sense strand is complementary to the antisense strand, wherein the antisense strand comprises a region complementary to part of a CFB RNA transcript, wherein each strand is about 15 to about 30 nucleotides in length, wherein the sense strand comprises sequence may be represented by formula (I) :
  • each N′ F represents a 2'-fluoro-modified nucleotide
  • each N′ N1 and N′ N2 independently represents a modified or unmodified nucleotide
  • each N′ L independently represents a modified or unmodified nucleotide but not a 2'-fluoro-modified nucleotide
  • m′and n′ are each independently an integer of 0 to 7.
  • N′ N1 and N′ N2 include only one 2'-Fluorine modified nucleotides.
  • N′ N1 independently represents a 2'-fluoro-modified nucleotide.
  • N′ N2 independently represents a 2'-fluoro-modified nucleotide.
  • the dsRNA agent includes a targeting group that is conjugated to the 5’-terminal end of the sense strand, preferably, the targeting group is any one selected from aforesaid GLO-1 through GLO-16 and GLS-1*through GLS-16*, more preferably, the targeting group is aforesaid GLS-15*.
  • the dsRNA agent includes a targeting group that is conjugated to the 3'-terminal end of the sense strand.
  • the antisense strand includes one inverted abasic residue at 3’-terminal end.
  • the sense strand includes one or two inverted abasic residues and/or one or two imann residues at 3’ or/and 5’ terminal end. In certain embodiments, each 3’ and 5’ terminal end of the sense strand independently includes an inverted abasic residue. In certain embodiments, each 3’ and 5’ terminal end of the sense strand independently includes an imann residue. In certain embodiments, the sense strand includes two inverted abasic residues at 3’ and 5’ terminal end and either residue at 3’ or 5’ terminal end is further conjugated to a targeting group, which preferably is aforesaid GLS-15*.
  • the sense strand includes two imann residues at 3’ and 5’ terminal end and either residue at 3’ or 5’ terminal end is further conjugated to a targeting group, which preferably is aforesaid GLS-15*.
  • each end of the sense strand includes one inverted abasic residue or imann residues at the 5' terminal end of the sense strand, wherein inverted abasic residue or imann residues is linked to an adjacent nucleotide via a phosphorothioate linkage to the 5' terminal end of the nucleotide sequence of the sense strand.
  • the sense strand further includes targeting group linked to an inverted abasic residue or an imann residue at the 5' terminal end of the sense strand, wherein targeting group is linked to an adjacent inverted abasic residue or imann via a phosphorothioate linkage, and optionally targeting group is N-acetyl-galactosamine (GalNAc) .
  • targeting group is linked to an adjacent inverted abasic residue or imann via a phosphorothioate linkage
  • optionally targeting group is N-acetyl-galactosamine (GalNAc) .
  • at the 5' terminal end of the sense strand includes one inverted abasic residue, wherein inverted abasic residue is linked to an adjacent nucleotide via a phosphorothioate linkage to the 5' terminal end of the nucleotide sequence of the sense strand.
  • the sense strand further includes targeting group linked to an inverted abasic residue at the 5' terminal end of the sense strand, wherein targeting group is linked to an adjacent inverted abasic residue via a phosphorothioate linkage, and optionally targeting group is N-acetyl-galactosamine (GalNAc) , the each strand is independently 21 nucleotides in length.
  • the antisense strand of the dsRNA agent is at least substantially complementary to any one of a target region of SEQ ID NO: 1 and is provided in any one of Tables 1-3.
  • a double-stranded ribonucleic acid (dsRNA) agent for inhibiting expression of CFB wherein the dsRNA agent including a sense strand and an antisense strand, wherein the sense strand is complementary to the antisense strand, wherein the antisense strand comprises a region complementary to a CFB RNA transcript, wherein each strand is about 18 to about 30 nucleotides in length, wherein the antisense strand comprises sequence may be represented by formula (II) :
  • each N F represents a 2'-fluoro-modified nucleotide
  • each N M1 , N M2 , N M3 , N M4 , N M5 , and N M6 independently represents a modified or unmodified nucleotide
  • each N L independently represents a modified or unmodified nucleotide but not a 2'-fluoro-modified nucleotide
  • n is an integer of 0 to 7.
  • N M1 , N M2 , N M3 , N M4 , N M5 , and N M6 have only three 2'-fluoro-modified nucleotides.
  • N M2 , N M3 and N M5 each independently represents a 2'-fluoro-modified nucleotide.
  • N M2 , N M4 and N M5 each independently represents a 2'-fluoro-modified nucleotide.
  • N M1 , N M3 and N M6 each independently represents a 2'-fluoro-modified nucleotide.
  • N M2 , N M3 and N M6 each independently represents a 2'-fluoro-modified nucleotide.
  • N M2 , N M4 and N M6 each independently represents a 2'-fluoro-modified nucleotide.
  • N M1 , N M3 and N M6 each independently represents a 2'-fluoro-modified nucleotide and N M5 represents an UNA modified nucleotide.
  • N M2 , N M3 and N M6 each independently represents a 2'-fluoro-modified nucleotide and N M5 represents an UNA modified nucleotide.
  • N M2 , N M4 and N M6 each independently represents a 2'-fluoro-modified nucleotide and N M5 represents an UNA modified nucleotide.
  • n is 1, or n is 2, or n is 3, or n is 5.
  • the antisense strand of the dsRNA agent is at least substantially complementary to any one of a target region of SEQ ID NO: 1 and is provided in any one of Tables 1-3.
  • a double-stranded ribonucleic acid (dsRNA) agent for inhibiting expression of CFB wherein the dsRNA agent including a sense strand and an antisense strand, wherein the sense strand and the antisense strand form a dsRNA duplex, wherein said sense strand is complementary to the antisense strand, wherein said antisense strand comprises a region of complementarity to a CFB RNA transcript, wherein the region of complementarity comprises at least 15 contiguous nucleotides, wherein the dsRNA duplex comprises represented by formula (III) :
  • each strand is about 18 to about 30 nucleotides in length
  • each N F and N′ F independently represents a 2'-fluoro-modified nucleotide
  • N M1 , N M2 , N M3 , N M4 , N M5 , N M6 , N′ N1 , and N′ N2 each independently represents a modified or unmodified nucleotide
  • each N L and N′ L independently represents a modified or unmodified nucleotide but not a 2'-fluoro-modified nucleotide, and m′, n′and n are each independently an integer of 0 to 7.
  • N M1 , N M2 , N M3 , N M4 , N M5 , and N M6 have only three 2'-fluoro-modified nucleotides, N′ N1 and N′ N2 include only one 2'-Fluorine modified nucleotides.
  • n 1, or n is 2, or n is 3, or n is 5.
  • N′ N1 independently represents a 2'-fluoro-modified nucleotide.
  • N′ N2 independently represents a 2'-fluoro-modified nucleotide.
  • N M2 , N M3 and N M5 each independently represents a 2'-fluoro-modified nucleotide.
  • N M2 , N M4 and N M5 each independently represents a 2'-fluoro-modified nucleotide.
  • N M1 , N M3 and N M6 each independently represents a 2'-fluoro-modified nucleotide.
  • N M2 , N M3 and N M6 each independently represents a 2'-fluoro-modified nucleotide.
  • N M2 , N M4 and N M6 each independently represents a 2'-fluoro-modified nucleotide.
  • N M1 , N M3 and N M6 each independently represents a 2'-fluoro-modified nucleotide and N M5 represents an UNA modified nucleotide.
  • N M2 , N M3 and N M6 each independently represents a 2'-fluoro-modified nucleotide and N M5 represents an UNA modified nucleotide.
  • N M2 , N M4 and N M6 each independently represents a 2'-fluoro-modified nucleotide and N M5 represents an UNA modified nucleotide.
  • the dsRNA agent includes a targeting group that is conjugated to the 5’-terminal end of the sense strand, preferably, the targeting group is any one selected from aforesaid GLO-1 through GLO-16 and GLS-1*through GLS-16*, more preferably, the targeting group is aforesaid GLS-15*.
  • the dsRNA agent includes a targeting group that is conjugated to the 5'-terminal end of the sense strand.
  • the antisense strand includes one inverted abasic residue at 3’-terminal end.
  • the sense strand includes one or two inverted abasic residues and/or one or two imann residues at 3’ or/and 5’ terminal end. In certain embodiments, each 3’ and 5’ terminal end of the sense strand independently includes an inverted abasic residue. In certain embodiments, each 3’ and 5’ terminal end of the sense strand independently includes an imann residue. In certain embodiments, the sense strand includes two inverted abasic residues at 3’ and 5’ terminal end and either residue at 3’ or 5’ terminal end is further conjugated to a targeting group, which preferably is aforesaid GLS-15*.
  • the sense strand includes two imann residues at 3’ and 5’ terminal end and either residue at 3’ or 5’ terminal end is further conjugated to a targeting group, which preferably is aforesaid GLS-15*.
  • the dsRNA agent has two blunt ends.
  • at least one strand includes a 3’ overhang of at least 1 nucleotide. In certain embodiments, at least one strand includes a 3’ overhang of at least 2 nucleotides.
  • each end of the sense strand includes one inverted abasic residue or imann residues at the 5' terminal end of the sense strand, wherein inverted abasic residue or imann residues is linked to an adjacent nucleotide via a phosphorothioate linkage to the 5' terminal end of the nucleotide sequence of the sense strand.
  • the sense strand further includes targeting group linked to an inverted abasic residue or an imann residue at the 5' terminal end of the sense strand, wherein targeting group is linked to an adjacent inverted abasic residue or imann via a phosphorothioate linkage, and optionally targeting group is N-acetyl-galactosamine (GalNAc) .
  • targeting group is linked to an adjacent inverted abasic residue or imann via a phosphorothioate linkage
  • optionally targeting group is N-acetyl-galactosamine (GalNAc) .
  • at the 5' terminal end of the sense strand includes one inverted abasic residue, wherein inverted abasic residue is linked to an adjacent nucleotide via a phosphorothioate linkage to the 5' terminal end of the nucleotide sequence of the sense strand.
  • the sense strand further includes targeting group linked to an inverted abasic residue at the 5' terminal end of the sense strand, wherein targeting group is linked to an adjacent inverted abasic residue via a phosphorothioate linkage, and optionally targeting group is N-acetyl-galactosamine (GalNAc) , the each strand is independently 21 nucleotides in length.
  • the antisense strand of the dsRNA agent is at least substantially complementary to any one of a target region of SEQ ID NO: 1 and is provided in any one of Tables 1-3.
  • a composition that includes any embodiment of the aforementioned dsRNA agent aspect of the invention.
  • the composition also includes a pharmaceutically acceptable carrier.
  • the composition also includes one or more additional therapeutic agents.
  • the composition is packaged in a kit, container, pack, dispenser, pre-filled syringe, or vial.
  • the composition is formulated for subcutaneous administration or is formulated for intravenous (IV) administration.
  • a cell that includes any embodiment of an aforementioned dsRNA agent aspect of the invention.
  • the cell is a mammalian cell, optionally a human cell.
  • a method of inhibiting the expression of a CFB gene in a cell including: (i) preparing a cell including an effective amount of any embodiment of the aforementioned dsRNA agent aspect of the invention or any embodiment of an aforementioned composition of the invention. In certain embodiments, the method also includes: (ii) maintaining the prepared cell for a time sufficient to obtain degradation of the mRNA transcript of a CFB gene, thereby inhibiting expression of the CFB gene in the cell.
  • the cell is in a subject and the dsRNA agent is administered to the subject subcutaneously. In some embodiments, the cell is in a subject and the dsRNA agent is administered to the subject by IV administration.
  • the method also includes assessing inhibition of the CFB gene, following the administration of the dsRNA agent to the subject, wherein a means for the assessing comprises: (i) determining one or more physiological characteristics of a CFB-associated disease or condition in the subject and (ii) comparing the determined physiological characteristic (s) to a baseline pre-treatment physiological characteristic of the CFB-associated disease or condition and/or to a control physiological characteristic of the CFB-associated disease or condition, wherein the comparison indicates one or more of a presence or absence of inhibition of expression of the CFB gene in the subject.
  • the physiological characteristic is one or more of: the CFB mRNA level and the CFB protein level.
  • a reduction in the expression of CFB may also be assessed indirectly by measuring a decrease in biological activity of CFB, e.g., or additional pathologies associated with elevated levels of CFB, preferably in the blood or in the kidneys, or over activation of the complement pathway, or additional therapeutic approaches where inhibition of CFB expression is desired.
  • a method of inhibiting expression of a CFB gene in a subject including administering to the subject an effective amount of an embodiment of the aforementioned dsRNA agent aspect of the invention or an embodiment of an aforementioned composition of the invention.
  • the dsRNA agent is administered to the subject subcutaneously.
  • the dsRNA agent is administered to the subject by IV administration.
  • the method also includes: assessing inhibition of the CFB gene, following the administration of the dsRNA agent, wherein a means for the assessing comprises: (i) determining one or more physiological characteristics of a CFB-associated disease or condition in the subject and (ii) comparing the determined physiological characteristic (s) to a baseline pre-treatment physiological characteristic of the CFB-associated disease or condition and/or to a control physiological characteristic of the CFB-associated disease or condition, wherein the comparison indicates one or more of a presence or absence of inhibition of expression of the CFB gene in the subject.
  • expression of the CFB gene can be assessed based on the level or change in level of any variable associated with CFB gene expression, such as CFB mRNA level, CFB protein level.
  • a reduction in the expression of CFB may also be assessed indirectly by measuring a decrease in biological activity of CFB, e.g., or additional pathologies associated with elevated levels of CFB, preferably in the blood or in the kidneys, or over activation of the complement pathway, or additional therapeutic approaches where inhibition of CFB expression is desired.
  • a method of treating a disease or condition associated with the presence of CFB protein including: administering to a subject an effective amount of an embodiment of any aforementioned dsRNA agent aspect of the invention or an embodiment of any aforementioned composition of the invention, to inhibit CFB gene expression.
  • the disease, disorder or condition associated with CFB is selected from the group consisting of: autoimmune diseases, complement system dysfunction including aberrant upregulation of complement components such as CFB, C3 glomerulopathy (C3G) , systemic lupus erythematosus (SLE) , Lupus Nephritis, Ig-mediated kidney pathologies such as IgA nephropathy and primary membranous nephropathy, nephropathy, diabetic nephropathy, polycystic kidney disease, membranous nephropathy, age-related macular degeneration (AMD) including dry AMD and geographic atrophy, typical or infectious hemolytic uremic syndrome (tHUS) , atypical hemolytic uremic syndrome (aHUS) , asthma, psoriasis, thrombotic microangiopathy, ischemia and reperfusion injury, paroxysmal nocturnal hemoglobinuria (PNH) , rheumatic disease, rheum
  • coli-related hemolytic uremic syndrome myasthenia gravis (MG) , neuromyelistis optica (NMO) , dense deposit disease, Coronary artery disease, dermatomyositis, Graves' disease, atherosclerosis, Alzheimer's disease, systemic inflammatory response sepsis, septic shock, spinal cord injury, glomerulonephritis, Hashimoto's thyroiditis, type I diabetes, psoriasis, pemphigus, autoimmune hemolytic anemia (AIHA) , cold agglutinin disease, humoral and vascular transplant rejection, graft dysfunction, myocardial infarction, sensitization towards a transplant, hyperlipidemia, and sepsis.
  • MG myasthenia gravis
  • NMO neuromyelistis optica
  • dense deposit disease Coronary artery disease
  • dermatomyositis CAD
  • Graves' disease atherosclerosis
  • Alzheimer's disease systemic inflammatory response seps
  • the method also includes: administering an additional therapeutic regimen to the subject.
  • the additional therapeutic regimen includes a treatment for the CFB-associated disease or condition.
  • the additional therapeutic regimen comprises: administering to the subject one or more CFB antisense polynucleotides of the invention, administering to the subject a non-CFB dsRNA therapeutic agent, and a behavioral modification in the subject.
  • the additional therapeutic agent is selected from the group consisting of an oligonucleotide, a small molecule, a monoclonal antibody, a polyclonal antibody and a peptide.
  • the additional therapeutic agent is an inhibitor of C5, such as an anticomplement component C5 antibody, or antigen-binding fragment thereof (e.g., eculizumab, ravulizumab-cwvz, or Polimab (REGN3918) ) or a C5 peptide inhibitor (e.g., zilucoplan) .
  • Eculizumab is a humanized monoclonal IgG2/4, kappa light chain antibody that specifically binds complement component C5 with high affinity and inhibits cleavage of C5 to C5a and C5b, thereby inhibiting the generation of the terminal complement complex C5b-9.
  • Ravulizumab-cwvz is a humanized IgG2/4 monoclonal antibody that specifically binds complement component C5 with high affinity and inhibits cleavage of C5 to C5a and C5b, thereby inhibiting the generation of the terminal complement complex C5b-9.
  • Pozelimab also known as H4H12166P, described in US20170355757
  • Zilucoplan is a synthetic, macrocyclic peptide that binds complement component 5 (C5) with sub-nanomolar affinity and allosterically inhibits its cleavage into C5a and C5b upon activation of the classical, alternative, or lectin pathways.
  • the additional therapeutic is a C3 peptide inhibitor, or analog thereof.
  • the C3 peptide inhibitor is compstatin.
  • Compstatin is a cyclic tridecapeptide with potent and selective C3 inhibitory activity.
  • the dsRNA agent is administered to the subject subcutaneously. In certain embodiments, the dsRNA agent is administered to the subject by IV administration. In some embodiments, the method also includes determining an efficacy of the administered double-stranded ribonucleic acid (dsRNA) agent in the subject.
  • dsRNA double-stranded ribonucleic acid
  • a means of determining an efficacy of the treatment in the subject comprises: (i) determining one or more physiological characteristics of the CFB-associated disease or condition in the subject and (ii) comparing the determined physiological characteristic (s) to a baseline pre-treatment physiological characteristic of the CFB-associated disease or condition wherein the comparison indicates one or more of a presence, absence, and level of efficacy of the administration of the double-stranded ribonucleic acid (dsRNA) agent to the subject.
  • dsRNA double-stranded ribonucleic acid
  • expression of the CFB gene can be assessed based on the level or change in level of any variable associated with CFB gene expression, such as CFB mRNA level, CFB protein level in the subject, or CH50 activity (a measure of total hemolytic complement) , AH50 (a measure the hemolytic activity of the alternate pathway of complement) , lactate dehydrogenase (LDH) (a measure of intravascular hemolysis) , hemoglobin levels; the level of any one or more of C3, C9, C5, C5a, C5b, and soluble C5b-9 complex.
  • CH50 activity a measure of total hemolytic complement
  • AH50 a measure the hemolytic activity of the alternate pathway of complement
  • LDH lactate dehydrogenase
  • hemoglobin levels the level of any one or more of C3, C9, C5, C5a, C5b, and soluble C5b-9 complex.
  • a method of decreasing a level of CFB protein in a subject compared to a baseline pre-treatment level of CFB protein in the subject including administering to the subject an effective amount of an embodiment of any aforementioned dsRNA agent of the invention or an embodiment of any aforementioned composition of the invention, to decrease the level of CFB gene expression.
  • the dsRNA agent is administered to the subject subcutaneously or is administered to the subject by IV administration.
  • a method of altering a physiological characteristic of a CFB-associated disease or condition in a subject compared to a baseline pre-treatment physiological characteristic of the CFB-associated disease or condition in the subject including administering to the subject an effective amount of an embodiment of any aforementioned dsRNA agent of the invention or an embodiment of any aforementioned composition of the invention, to alter the physiological characteristic of the CFB-associated disease or condition in the subject.
  • the dsRNA agent is administered to the subject subcutaneously or is administered to the subject by IV administration.
  • the physiological characteristic and symptoms are one or more of: CFB mRNA level, CFB protein level in the subject, or CH50 activity (a measure of total hemolytic complement) , AH50 (a measure the hemolytic activity of the alternate pathway of complement) , lactate dehydrogenase (LDH) (a measure of intravascular hemolysis) , hemoglobin levels; the level of any one or more of C3, C9, C5, C5a, C5b, and soluble C5b-9 complex.
  • CH50 activity a measure of total hemolytic complement
  • AH50 a measure the hemolytic activity of the alternate pathway of complement
  • LDH lactate dehydrogenase
  • hemoglobin levels the level of any one or more of C3, C9, C5, C5a, C5b, and soluble C5b-9 complex.
  • the aforementioned dsRNA agent for use in a method of treating a disease or condition associated with the presence of CFB protein.
  • the disease or condition is one or more of: autoimmune diseases, complement system dysfunction including aberrant upregulation of complement components such as CFB, C3 glomerulopathy (C3G) , systemic lupus erythematosus (SLE) , Lupus Nephritis, Ig-mediated kidney pathologies such as IgA nephropathy and primary membranous nephropathy, nephropathy, diabetic nephropathy, polycystic kidney disease, membranous nephropathy, age-related macular degeneration (AMD) including dry AMD and geographic atrophy, typical or infectious hemolytic uremic syndrome (tHUS) , atypical hemolytic uremic syndrome (aHUS) , asthma, psoriasis, thrombotic microangiopathy, ischemia and reperfusion
  • C3G C3 glomerulopathy
  • coli-related hemolytic uremic syndrome myasthenia gravis (MG) , neuromyelistis optica (NMO) , dense deposit disease, Coronary artery disease, dermatomyositis, Graves' disease, atherosclerosis, Alzheimer's disease, systemic inflammatory response sepsis, septic shock, spinal cord injury, glomerulonephritis, Hashimoto's thyroiditis, type I diabetes, psoriasis, pemphigus, autoimmune hemolytic anemia (AIHA) , cold agglutinin disease, humoral and vascular transplant rejection, graft dysfunction, myocardial infarction, sensitization towards a transplant, hyperlipidemia, and sepsis.
  • MG myasthenia gravis
  • NMO neuromyelistis optica
  • dense deposit disease Coronary artery disease
  • dermatomyositis CAD
  • Graves' disease atherosclerosis
  • Alzheimer's disease systemic inflammatory response seps
  • an antisense polynucleotide agent for inhibiting expression of CFB protein including from 10 to 30 contiguous nucleotides, wherein at least one of the contiguous nucleotides is a modified nucleotide, and wherein the nucleotide sequence of the agent is about 80%complementary over its entire length to the equivalent region of the nucleotide sequence of SEQ ID NO: 1.
  • the equivalent region is any one of the target regions of SEQ ID NO: 1 and the complementary sequence is one provided in one of Tables 1-3.
  • the antisense polynucleotide agent includes one of the antisense sequences provided in one of Tables 1-3.
  • compositions including an embodiment of any aforementioned antisense polynucleotide agents is provided.
  • the composition also includes a pharmaceutically acceptable carrier.
  • the composition also includes one or more additional therapeutic agents for treatment of a CFB-associated disease or condition.
  • the composition is packaged in a kit, container, pack, dispenser, pre-filled syringe, or vial.
  • the composition is formulated for subcutaneous or IV administration.
  • a cell that includes an embodiment of any of the aforementioned antisense polynucleotide agents is provided.
  • the cell is a mammalian cell, optionally a human cell.
  • a method of inhibiting the expression of a CFB gene in a cell including: (i) preparing a cell including an effective amount of an embodiment of any aforementioned antisense polynucleotide agents. In some embodiments, the method also includes (ii) maintaining the cell prepared in (i) for a time sufficient to obtain degradation of the mRNA transcript of a CFB gene, thereby inhibiting expression of the CFB gene in the cell.
  • a method of inhibiting expression of a CFB gene in a subject including administering to the subject an effective amount of an embodiment of any of the aforementioned antisense polynucleotide agent.
  • a method of treating a disease or condition associated with the presence of CFB protein including administering to a subject an effective amount of an embodiment of any of the aforementioned antisense polynucleotide agents or an embodiment of any aforementioned composition of the invention, to inhibit CFB gene expression.
  • the disease or condition is one or more of: autoimmune diseases, complement system dysfunction including aberrant upregulation of complement components such as CFB, C3 glomerulopathy (C3G) , systemic lupus erythematosus (SLE) , Lupus Nephritis, Ig-mediated kidney pathologies such as IgA nephropathy and primary membranous nephropathy, nephropathy, diabetic nephropathy, polycystic kidney disease, membranous nephropathy, age-related macular degeneration (AMD) including dry AMD and geographic atrophy, typical or infectious hemolytic uremic syndrome (tHUS) , atypical hemolytic uremic syndrome (aHUS) , asthma, psoriasis, thrombotic microangiopathy, ischemia and reperfusion injury, paroxysmal nocturnal hemoglobinuria (PNH) , rheumatic disease, rheumatoid arthritis, multiple s
  • CFB
  • coli-related hemolytic uremic syndrome myasthenia gravis (MG) , neuromyelistis optica (NMO) , dense deposit disease, Coronary artery disease, dermatomyositis, Graves' disease, atherosclerosis, Alzheimer's disease, systemic inflammatory response sepsis, septic shock, spinal cord injury, glomerulonephritis, Hashimoto's thyroiditis, type I diabetes, psoriasis, pemphigus, autoimmune hemolytic anemia (AIHA) , cold agglutinin disease, humoral and vascular transplant rejection, graft dysfunction, myocardial infarction, sensitization towards a transplant, hyperlipidemia, and sepsis.
  • MG myasthenia gravis
  • NMO neuromyelistis optica
  • dense deposit disease Coronary artery disease
  • dermatomyositis CAD
  • Graves' disease atherosclerosis
  • Alzheimer's disease systemic inflammatory response seps
  • a method of decreasing a level of CFB protein in a subject compared to a baseline pre-treatment level of CFB protein in the subject including administering to the subject an effective amount of an embodiment of any of the aforementioned antisense polynucleotide agents or an embodiment of any aforementioned composition of the invention, to decrease the level of CFB gene expression.
  • the antisense polynucleotide agent is administered to the subject subcutaneously or by IV administration.
  • an antisense polynucleotide agent for inhibiting expression of CFB gene including from 10 to 30 contiguous nucleotides, wherein at least one of the contiguous nucleotides is a modified nucleotide, and wherein the nucleotide sequence of the agent is about 80%or about 85%complementary over its entire length to the equivalent region of the nucleotide sequence of SEQ ID NO: 1.
  • a method of altering a physiological characteristic of a CFB-associated disease or condition in a subject compared to a baseline pre-treatment physiological characteristic of the CFB-associated disease or condition in the subject including administering to the subject an effective amount of an embodiment of any of the aforementioned antisense polynucleotide agents or an embodiment of any aforementioned composition of the invention, to alter the physiological characteristic of the CFB disease or condition in the subject.
  • the antisense polynucleotide agent is administered to the subject subcutaneously or by IV administration.
  • the physiological characteristic and symptoms are one or more of: CFB mRNA level, CFB protein level in the subject, or CH50 activity (a measure of total hemolytic complement) , AH50 (a measure the hemolytic activity of the alternate pathway of complement) , lactate dehydrogenase (LDH) (a measure of intravascular hemolysis) , hemoglobin levels; the level of any one or more of C3, C9, C5, C5a, C5b, and soluble C5b-9 complex.
  • CH50 activity a measure of total hemolytic complement
  • AH50 a measure the hemolytic activity of the alternate pathway of complement
  • LDH lactate dehydrogenase
  • hemoglobin levels the level of any one or more of C3, C9, C5, C5a, C5b, and soluble C5b-9 complex.
  • SEQ ID NO: 1 and SEQ ID NO: 2 are Homo sapiens CFB mRNA [NCBI Reference Sequence: NM_001710.6] .
  • SEQ ID NO: 3 and SEQ ID NO: 4 are Macaca mulatta (Rhesus monkey) CFB mRNA [NCBI Reference Sequence: XM_015136029.2] .
  • SEQ ID NO: 5 and SEQ ID NO: 6 are Mus musculus (isoform 1) CFB mRNA [NCBI Reference Sequence: NM_008198.3] .
  • SEQ ID Nos: 7-468, 1489-1595 are shown in Table 1 and are sense strand sequences.
  • SEQ ID Nos: 469-930, 1596-1702 are shown in Table 1 and are antisense strand sequences.
  • SEQ ID Nos: 931-1392, 1703-1810 are shown in Table 2 with chemical modifications.
  • a delivery molecule is indicated as “GLX-__” at the 3’ end or 5’ end of each sense strand.
  • the invention in part, includes RNAi agents, for example, though not limited to double stranded (ds) RNAi agents, which are capable of inhibiting CFB gene expression.
  • the invention in part also includes compositions comprising CFB RNAi agents and methods of use of the compositions.
  • CFB RNAi agents disclosed herein may be attached to delivery compounds for delivery to cells, including to hepatocytes.
  • Pharmaceutical compositions of the invention may include at least one dsRNA CFB agent and a delivery compound.
  • the delivery compound is a GalNAc-containing delivery compound.
  • CFB RNAi agents delivered to cells are capable of inhibiting CFB gene expression, thereby reducing activity in the cell of the CFB protein product of the gene.
  • dsRNAi agents of the invention can be used to treat CFB-associated diseases and conditions.
  • reducing CFB expression in a cell or subject treats a disease or condition associated with CFB expression in the cell or subject, respectively.
  • diseases and conditions that may be treated by reducing CFB activity are: alleviation or amelioration of one or more symptoms associated with unwanted or excessive CFB expression, CH50 activity (a measure of total hemolytic complement) , AH50 (a measure the hemolytic activity of the alternate pathway of complement) , lactate dehydrogenase (LDH) (a measure of intravascular hemolysis) , hemoglobin levels; the level of any one or more of C3, C9, C5, C5a, C5b, and soluble C5b-9 complex.
  • “Treatment” can also mean prolonging survival as compared to expected survival in the absence of treatment.
  • G, " C, “ “A” and “U” each generally stand for a nucleotide that contains guanine, cytosine, adenine, and uracil as a base, respectively.
  • ribonucleotide or “nucleotide” can also refer to a modified nucleotide, as further detailed below, or a surrogate replacement moiety.
  • guanine, cytosine, adenine, and uracil may be replaced by other moieties without substantially altering the base pairing properties of an oligonucleotide comprising a nucleotide bearing such replacement moiety.
  • nucleotide comprising inosine as its base may base pair with nucleotides containing adenine, cytosine, or uracil.
  • nucleotides containing uracil, guanine, or adenine may be replaced in the nucleotide sequences of the invention by a nucleotide containing, for example, inosine. Sequences comprising such replacement moieties are embodiments of the invention.
  • Complement Factor B used interchangeably with the term “factor B” or “CFB” refers to the naturally occurring gene that encodes a Complement Factor B protein from any vertebrate or mammalian source, including, but not limited to, human, bovine, chicken, rodent, mouse, rat, porcine, ovine, primate, monkey, and guinea pig, unless specified otherwise.
  • the term also refers to fragments and variants of native CFB that maintain at least one in vivo or in vitro activity of a native CFB.
  • the amino acid and complete coding sequences of the reference sequence of the human CFB gene may be found in, for example, GenBank Ref Seq Accession No.
  • NM_001710.6 (SEQ ID NO: 1 and SEQ ID NO: 2) .
  • Macaca mulatta (Rhesus monkey) CFB mRNA GenBank Ref Seq Accession No. XM_015136029.2 (SEQ ID NO: 3 and SEQ ID NO: 4) , Mus musculus (isoform 1) NM_008198.3 (SEQ ID NO: 5 and SEQ ID NO: 6) .
  • Additional examples of CFB mRNA sequences are readily available using publicly available databases, e.g., GenBank, UniProt, Ensembl and OMIM.
  • RNAi CFB single-stranded
  • siRNA dsRNA agents
  • RNAi refers to an agent that comprises RNA and mediates targeted cleavage of an RNA transcript via an RNA-induced silencing complex (RISC) pathway.
  • RISC RNA-induced silencing complex
  • an RNAi a target region refers to a contiguous portion of the nucleotide sequence of an mRNA molecule formed during the transcription of a gene, including messenger RNA (mRNA) that is a product of RNA processing of a primary transcription product.
  • mRNA messenger RNA
  • the target portion of the sequence will be at least long enough to serve as a substrate for RNAi-directed cleavage at or near that portion.
  • a target sequence may be from 8-30 nucleotides long (inclusive) , from 10 -30 nucleotides long (inclusive) , from 12 -25 nucleotides long (inclusive) , from 15 -23 nucleotides long (inclusive) , from 16 -23 nucleotides long (inclusive) , or from 18 –23 nucleotides long (inclusive) , including all shorter lengths within each stated range.
  • a target sequence is 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, or 26 nucleotides long.
  • a target sequence is between 9 and 26 nucleotides long (inclusive) , including all sub-ranges and integers there between.
  • a target sequence is 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleotides long, with the sequence fully or at least substantially complementary to at least part of an RNA transcript of a CFB gene.
  • Some aspects of the invention include pharmaceutical compositions comprising one or more CFB dsRNA agents and a pharmaceutically acceptable carrier.
  • a CFB RNAi as described herein inhibits expression of CFB protein.
  • a “dsRNA agent” means a composition that contains an RNA or RNA-like (e.g., chemically modified RNA) oligonucleotide molecule that is capable of degrading or inhibiting translation of messenger RNA (mRNA) transcripts of a target mRNA in a sequence specific manner.
  • dsRNA agents of the invention may operate through the RNA interference mechanism (i.e., inducing RNA interference through interaction with the RNA interference pathway machinery (RNA-induced silencing complex or RISC) of mammalian cells) , or by any alternative mechanism (s) or pathway (s) .
  • DsRNA agents disclosed herein are comprised of a sense strand and an antisense strand, and include, but are not limited to: short interfering RNAs (siRNAs) , RNAi agents, micro RNAs (miRNAs) , short hairpin RNAs (shRNA) , and dicer substrates.
  • the antisense strand of the dsRNA agents described herein is at least partially complementary to the mRNA being targeted. It is understood in the art that different lengths of dsRNA duplex structure can be used to inhibit target gene expression.
  • dsRNAs having a duplex structure of 19, 20, 21, 22, and 23 base pairs are known to be effective to induce RNA interference (Elbashir et al., EMBO 2001, 20: 6877-6888) . It is also known in the art that shorter or longer RNA duplex structures are also effective to induce RNA interference.
  • the sense strand and the antisense strand may be the same length or different lengths.
  • each strand is no more than 40 nucleotides in length.
  • each strand is no more than 30 nucleotides in length.
  • each strand is no more than 25 nucleotides in length.
  • each strand is no more than 23 nucleotides in length.
  • each strand is no more than 21 nucleotides in length.
  • the sense and antisense strands of the RNAi agents can each be 15 to 49 nucleotides in length.
  • the antisense strand is independently 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleotides in length.
  • the length of the sense strand is independently 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, or 49 nucleotides.
  • the sense strand and the antisense strand are both 21 nucleotides in length.
  • the sense strand is complementary or substantially complementary to the antisense strand, and the region of complementarity is between 15 and 23 nucleotides in length.
  • the region of complementarity is 19-21 nucleotides in length.
  • the region of complementarity is 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleotides in length.
  • CFB dsRNAs in certain embodiments of the invention can include at least one strand of a length of minimally 21 nt or may have shorter duplexes based on one of the sequences set forth in any one of Tables 1-3, but minus 1, 2, 3, or 4 nucleotides on one or both ends may also be effective as compared to the dsRNAs set forth in Tables 1-3, respectively.
  • CFB dsRNA agents may have a partial sequence of at least 15, 16, 17, 18, 19, 20, or more contiguous nucleotides from one or more sequences of Tables 1-3, and differ in their ability to inhibit the expression of a CFB gene by not more than 5%, 10%, 15%, 20%, 25%, or 30%from the level of inhibition resulting from a dsRNA comprising the full sequence.
  • a sense sequence, an antisense sequence and a duplex disclosed in Tables 1-3 may be referred to herein as a “parent” sequence, meaning that the sequences disclosed in Tables 1-3 may be modified, shorten, lengthened, include substitutions, etc.
  • Sense and antisense strands included in a dsRNA of the invention are independently selected.
  • independently selected means each of two or more like elements can be selected independent of the selection of the other elements. For example, though not intended to be limiting, in preparing a dsRNA of the invention, one may select the “elements” of the two strands to include in the duplex.
  • the sense sequence may be SEQ ID NO: 932 (shown in Table 2) and the other selected element, the antisense sequence, may be SEQ ID NO: 1163, or may be SEQ ID NO: 1163 that is modified, shortened, lengthened, and/or includes 1, 2, or 3 substitutions as compared to its parent sequence SEQ ID NO: 1163.
  • a duplex of the invention need not include both sense and antisense sequences shown as paired in duplexes in Tables 1-3. Each sense and antisense strand sequence in the tables is immediately followed by its SEQ ID NO.
  • compositions and methods of the invention comprise a single-strand RNA in a composition and/or administered to a subject.
  • an antisense strand such as one listed in any one of Tables 1-3 may be a composition or in a composition administered to a subject to reduce CFB polypeptide activity and/or expression of CFB gene in the subject.
  • Tables 1 shows certain CFB dsRNA agent antisense strand and sense strand core stretch base sequences.
  • a single-strand antisense molecule that may be included in certain compositions and/or administered in certain methods of the invention are referred to herein as a “single-strand antisense agent” or an “antisense polynucleotide agent” .
  • a single-strand sense molecule that may be included in certain compositions and/or administered in certain methods of the invention are referred to herein as a “single-strand sense agent” or a “sense polynucleotide agent” .
  • the term “base sequence” is used herein in reference to a polynucleotide sequence without chemical modifications or delivery compounds.
  • the sense strand GACAAUGUGAGUGAUGAGAUA (SEQ ID NO: 22) shown in Table 1 is the base sequence for SEQ ID NO: 946 in Table 2 and for SEQ ID NO: 1393 in Table 3, with SEQ ID NO: 946 and SEQ ID NO: 1393 shown with their chemical modifications and a delivery compound. Sequences disclosed herein may be assigned identifiers.
  • a single-stranded sense sequence may be identified with a “Sense strand SS#” ; a single stranded antisense sequence may be identified with an “Antisense strand AS#” and a duplex that includes a sense strand and an antisense strand may be identified with a “Duplex AD#/AV#” .
  • Table 1 includes sense and antisense strands and provides the identification number of duplexes formed from the sense and antisense strand on the same line in Table 1.
  • an antisense sequence includes nucleobase u or nucleobase a in position 1 of the antisense sequence.
  • an antisense sequence includes nucleobase u in position 1 of the antisense sequence.
  • the term “matching position” in a sense and an antisense strand are the positions in each strand that “pair” when the two strands are duplexed strands.
  • nucleobase in position 1 of the sense strand and position 21 in the antisense strand are in “matching positions” .
  • nucleobase 2 of the sense strand and position 22 of the antisense strand are in matching positions.
  • nucleobase in position 1 of the sense strand and nucleobase 18 in the antisense strand are in matching positions
  • nucleobase 4 in the sense strand and nucleobase 15 in the antisense strand are in matching positions.
  • the final column in Table 1 indicates a Duplex AV# for a duplex that includes the sense and antisense sequences in the same table row.
  • Table 1 discloses the duplex assigned Duplex AV02358. um, which includes sense strand SEQ ID NO: 7 and antisense strand SEQ ID NO: 469.
  • each row in Table 1 identifies a duplex of the invention, each comprising the sense and antisense sequences shown in the same row, with the assigned identifier for each duplex shown in the final column in the row.
  • an RNAi agent comprising a polynucleotide sequence shown in any one of Tables 1-3 is administered to a subject.
  • an RNAi agent administered to a subject comprises is a duplex comprising at least one of the base sequences set forth in Table 1, including 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, or 24 sequence modifications.
  • an RNAi agent comprising a polynucleotide sequence shown in any one of Tables 1-3 is attached to a delivery molecule, a non-limiting example of which is a delivery compound comprising a GalNAc compound, or a GLS-15*compound.
  • a dsRNA (also referred to herein as a “duplex” ) is one disclosed in one of Tables 1-3.
  • Tables 1-3 Each row in Tables 1-3 discloses a duplex comprising the sequence of the sense strand and the sequence of the antisense strand in that table row.
  • a duplex of the invention may include sense and antisense sequences shown in Tables 1-3, that differ by zero, one, two, or three nucleotides shown in a sequence shown in Tables 1-3.
  • an antisense strand in a duplex of the invention may be SEQ ID NO: 484, 490, 497, 665, or 667 with zero, one, two, or three different nucleotides than those in SEQ ID484, 490, 497, 665, or 667 respectively.
  • a dsRNA of the invention may comprise a sense strand and an antisense strand of a duplex disclosed in a row in Tables 1-3.
  • one or both of the selected sense and antisense strand in the dsRNA may include sequences shown in Tables 1-3 but with one or both of the sense and antisense sequences including 1, 2, 3, or more nucleobase substitutions from the parent sequence.
  • the selected sequences may in some embodiments be longer or shorter than their parent sequence.
  • dsRNA agents included in the invention can but need not include exact sequences of the sense and antisense pairs disclosed as duplexes in Tables 1-3.
  • a dsRNA agent comprises a sense strand and an antisense strand, nucleotide positions 2 to 18 in the antisense strand comprising a region of complementarity to a CFB RNA transcript, wherein the region of complementarity comprises at least 15 contiguous nucleotides that differ by 0, 1, 2, or 3 nucleotides from one of the antisense sequences listed in one of Tables 1-3, and optionally comprising a targeting ligand.
  • the region of complementarity to the CFB RNA transcript comprises at least 15, 16, 17, 18, or 19 contiguous nucleotides that differ by no more than 3 nucleotides from one of the antisense sequences listed in one of Tables 1-3.
  • the antisense strand of the dsRNA is at least substantially complementary to any one of a target region of SEQ ID NO: 1 and is provided in any one of Tables 1-3.
  • an antisense strand of a dsRNA agent of the invention is fully complementary to any one of a target region of SEQ ID NO: 1 and is provided in any one of Tables 1-3.
  • a dsRNA agent includes a sense strand sequence set forth in any one of Tables 1-3, and the sense strand sequence is at least substantially complementary to the antisense strand sequence in the dsRNA agent.
  • a dsRNA agent of the invention comprises a sense strand sequence set forth in any one of Tables 1-3, and the sense strand sequence is fully complementary to the antisense strand sequence in the dsRNA agent.
  • a dsRNA agent of the invention comprises an antisense strand sequence set forth in any one of Tables 1-3.
  • Some embodiments of a dsRNA agent of the invention comprises the sense and antisense sequences disclosed as duplex in any of Tables 1-3. As described herein, it will be understood that the sense and antisense strands in a duplex of the invention may be independently selected.
  • mismatches are tolerated for efficacy in dsRNA, especially the mismatches are within terminal region of dsRNA.
  • Certain mismatches tolerate better, for example mismatches with wobble base pairs G: U and A: C are tolerated better for efficacy (Du et el., A systematic analysis of the silencing effects of an active siRNA at all single-nucleotide mismatched target sites.
  • a CFB dsRNA agent may contain one or more mismatches to the CFB target sequence.
  • CFB dsRNA agent of the invention includes no mismatches.
  • CFB dsRNA agent of the invention includes no more than 1 mismatch.
  • CFB dsRNA agent of the invention includes no more than 2 mismatches.
  • CFB dsRNA agent of the invention includes no more than 3 mismatches.
  • an antisense strand of a CFB dsRNA agent contains mismatches to a CFB target sequence that are not located in the center of the region of complementarity.
  • the antisense strand of the CFB dsRNA agent includes 1, 2, 3, 4, or more mismatches that are within the last 5, 4, 3, 2, or 1 nucleotide from one or both of the 5' or 3' end of the region of complementarity.
  • the term “complementary, ” when used to describe a first nucleotide sequence (e.g., CFB dsRNA agent sense strand or targeted CFB mRNA) in relation to a second nucleotide sequence (e.g., CFB dsRNA agent antisense strand or a single-stranded antisense polynucleotide) means the ability of an oligonucleotide or polynucleotide including the first nucleotide sequence to hybridize [form base pair hydrogen bonds under mammalian physiological conditions (or similar conditions in vitro) ] and form a duplex or double helical structure under certain conditions with an oligonucleotide or polynucleotide including the second nucleotide sequence.
  • Complementary sequences include Watson-Crick base pairs or non-Watson-Crick base pairs and include natural or modified nucleotides or nucleotide mimics, at least to the extent that the above hybridization requirements are fulfilled. Sequence identity or complementarity is independent of modification.
  • Complementary sequences for example, within a CFB dsRNA as described herein, include base-pairing of the oligonucleotide or polynucleotide comprising a first nucleotide sequence to an oligonucleotide or polynucleotide comprising a second nucleotide sequence over the entire length of one or both nucleotide sequences.
  • Such sequences can be referred to as “fully complementary” with respect to each other herein. It will be understood that in embodiments when two oligonucleotides are designed to form, upon hybridization, one or more single stranded overhangs, such overhangs are not regarded herein as mismatches with regard to the determination of complementarity.
  • a CFB dsRNA agent comprising one oligonucleotide 19 nucleotides in length and another oligonucleotide 20 nucleotides in length, wherein the longer oligonucleotide comprises a sequence of 19 nucleotides that is fully complementary to the shorter oligonucleotide, can yet be referred to as “fully complementary” for the purposes described herein.
  • “fully complementary” means that all (100%) of the bases in a contiguous sequence of a first polynucleotide will hybridize with the same number of bases in a contiguous sequence of a second polynucleotide.
  • the contiguous sequence may comprise all or a part of a first or second nucleotide sequence.
  • substantially complementary means that in a hybridized pair of nucleobase sequences, at least about 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%, but not all, of the bases in a contiguous sequence of a first polynucleotide will hybridize with the same number of bases in a contiguous sequence of a second polynucleotide.
  • substantially complementary can be used in reference to a first sequence with respect to a second sequence if the two sequences include one or more, for example at least 1, 2, 3, 4, or 5 mismatched base pairs upon hybridization for a duplex up to 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29 or 30 base pairs (bp) , while retaining the ability to hybridize under the conditions most relevant to their ultimate application, e.g., inhibition of CFB gene expression via a RISC pathway.
  • partially complementary may be used herein in reference to a hybridized pair of nucleobase sequences, in which at least 75%, but not all, of the bases in a contiguous sequence of a first polynucleotide will hybridize with the same number of bases in a contiguous sequence of a second polynucleotide.
  • “partially complementary” means at least 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%of the bases in a contiguous sequence of a first polynucleotide will hybridize with the same number of bases in a contiguous sequence of a second polynucleotide.
  • complementary, ” “fully complementary, ” “substantially complementary, ” and “partially complimentary” are used herein in reference to the base matching between the sense strand and the antisense strand of a CFB dsRNA agent, between the antisense strand of a CFB dsRNA agent and a sequence of a target CFB mRNA, or between a single-stranded antisense oligonucleotide and a sequence of a target CFB mRNA.
  • antisense strand of a CFB dsRNA agent may refer to the same sequence of an “CFB antisense polynucleotide agent” .
  • nucleic acid sequence As used herein, the term “substantially identical” or “substantial identity” used in reference to a nucleic acid sequence means a nucleic acid sequence comprising a sequence with at least about 85%sequence identity or more, preferably at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, compared to a reference sequence. Percentage of sequence identity is determined by comparing two optimally aligned sequences over a comparison window.
  • the percentage is calculated by determining the number of positions at which the identical nucleic acid base occurs in both sequences to yield the number of matched positions, dividing the number of matched positions by the total number of positions in the window of comparison and multiplying the result by 100 to yield the percentage of sequence identity.
  • the inventions disclosed herein encompasses nucleotide sequences substantially identical to those disclosed herein. e.g., in Tables 1-3. In some embodiments, the sequences disclosed herein are exactly identical, or at least about 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%percent identical to those disclosed herein, e.g., in Tables 1-3.
  • strand comprising a sequence means an oligonucleotide comprising a chain of nucleotides that is described by the sequence referred to using the standard nucleotide nomenclature.
  • double-stranded RNA or “dsRNA, ” as used herein, refers to an RNAi that includes an RNA molecule or complex of molecules having a hybridized duplex region comprising two anti-parallel and substantially or fully complementary nucleic acid strands, which are referred to as having “sense” and “antisense” orientations with respect to a target CFB RNA.
  • the duplex region can be of any length that permits specific degradation of a desired target CFB RNA through a RISC pathway but will typically range from 9 to 30 base pairs in length, e.g., 15-30 base pairs in length.
  • the duplex can be any length in this range, for example, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30, and any sub-range therein between, including, but not limited to 15-30 base pairs, 15-26 base pairs, 15-23 base pairs, 15-22 base pairs, 15-21 base pairs, 15-20 base pairs, 15-19 base pairs, 15-18 base pairs, 15-17 base pairs, 18-30 base pairs, 18-26 base pairs, 18-23 base pairs, 18-22 base pairs, 18-21 base pairs, 18-20 base pairs, 19-30 base pairs, 19-26 base pairs, 19-23 base pairs, 19-22 base pairs, 19-21 base pairs, 19-20 base pairs, 20-30 base pairs, 20-26 base pairs, 20-25 base pairs, 20-24 base pairs, 20-23 base
  • CFB dsRNA agents generated in the cell by processing with Dicer and similar enzymes are generally in the range of 19-22 base pairs in length.
  • One strand of the duplex region of a CFB dsDNA agent comprises a sequence that is substantially complementary to a region of a target CFB RNA.
  • the two strands forming the duplex structure can be from a single RNA molecule having at least one self-complementary region, or can be formed from two or more separate RNA molecules.
  • the molecule can have a duplex region separated by a single stranded chain of nucleotides (herein referred to as a “hairpin loop” ) between the 3'-end of one strand and the 5'-end of the respective other strand forming the duplex structure.
  • a hairpin look comprises at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, or more unpaired nucleotides.
  • RNA is also used herein to refer to a dsRNA agent as described herein.
  • a CFB dsRNA agent may include a sense and antisense sequence that have no-unpaired nucleotides or nucleotide analogs at one or both terminal ends of the dsRNA agent.
  • An end with no unpaired nucleotides is referred to as a “blunt end” and as having no nucleotide overhang. If both ends of a dsRNA agent are blunt, the dsRNA is referred to as “blunt ended.
  • a first end of a dsRNA agent is blunt, in some embodiments a second end of a dsRNA agent is blunt, and in certain embodiments of the invention, both ends of a CFB dsRNA agent are blunt.
  • the dsRNA does not have one or two blunt ends.
  • a dsRNA can comprise an overhang of at least 1, 2, 3, 4, 5, 6, or more nucleotides.
  • a nucleotide overhang can comprise or consist of a nucleotide/nucleoside analog, including a deoxynucleotide/nucleoside.
  • nucleotide overhang is on a sense strand of a dsRNA agent, on an antisense strand of a dsRNA agent, or on both ends of a dsRNA agent and nucleotide (s) of an overhang can be present on the 5' end, 3' end or both ends of either an antisense or sense strand of a dsRNA.
  • nucleotides in an overhang is replaced with a nucleoside thiophosphate.
  • antisense strand or “guide strand” refers to the strand of a CFB dsRNA agent that includes a region that is substantially complementary to a CFB target sequence.
  • sense strand, ” or “passenger strand” refers to the strand of a CFB dsRNA agent that includes a region that is substantially complementary to a region of the antisense strand of the CFB dsRNA agent.
  • the RNA of a CFB RNAi agent is chemically modified to enhance stability and/or one or more other beneficial characteristics.
  • Nucleic acids in certain embodiments of the invention may be synthesized and/or modified by methods well established in the art, for example, those described in “Current protocols in Nucleic Acid Chemistry, " Beaucage, S.L. et al. (Eds. ) , John Wiley & Sons, Inc., New York, N.Y., USA, which is incorporated herein by reference.
  • Modifications that can be present in certain embodiments of CFB dsRNA agents of the invention include, for example, (a) end modifications, e.g., 5' end modifications (phosphorylation, conjugation, inverted linkages, etc. ) 3' end modifications (conjugation, DNA nucleotides, inverted linkages, etc.
  • RNA compounds useful in certain embodiments of CFB dsRNA agents, CFB antisense polynucleotides, and CFB sense polynucleotides of the invention include, but are not limited to RNAs comprising modified backbones or no natural internucleoside linkages.
  • an RNA having a modified backbone may not have a phosphorus atom in the backbone.
  • RNAs that do not have a phosphorus atom in their internucleoside backbone may be referred to as oligonucleosides.
  • a modified RNA has a phosphorus atom in its internucleoside backbone.
  • RNA molecule or “RNA” or “ribonucleic acid molecule” encompasses not only RNA molecules as expressed or found in nature, but also analogs and derivatives of RNA comprising one or more ribonucleotide/ribonucleoside analogs or derivatives as described herein or as known in the art.
  • ribonucleoside and “ribonucleotide” may be used interchangeably herein.
  • An RNA molecule can be modified in the nucleobase structure or in the ribose-phosphate backbone structure, e.g., as described herein below, and molecules comprising ribonucleoside analogs or derivatives must retain the ability to form a duplex.
  • an RNA molecule can also include at least one modified ribonucleoside including but not limited to a 2'-O-methyl modified nucleoside, a nucleoside comprising a 5' phosphorothioate group, a terminal nucleoside linked to a cholesteryl derivative or dodecanoic acid bisdecylamide group, a locked nucleoside, an abasic nucleoside, a 2'-deoxy-2'-fluoro modified nucleoside, a 2'-amino-modified nucleoside, 2'-alkyl-modified nucleoside, morpholino nucleoside, a phosphoramidate or a non-natural base comprising nucleoside, or any combination thereof.
  • a 2'-O-methyl modified nucleoside a nucleoside comprising a 5' phosphorothioate group, a terminal nucleoside linked to a cholesteryl derivative or dodecanoic acid bisdec
  • an RNA molecule comprises at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, or up to the full length of the CFB dsRNA agent molecule’s ribonucleosides that are modified ribonucleosides.
  • the modifications need not be the same for each of such a plurality of modified ribonucleosides in an RNA molecule.
  • DsRNA agents, CFB antisense polynucleotides, and/or CFB sense polynucleotides of the invention may, in some embodiments comprise one or more independently selected modified nucleotide and/or one or more independently selected non-phosphodiester linkage.
  • independently selected used in reference to a selected element, such as a modified nucleotide, non-phosphodiester linkage, etc., means that two or more selected elements can but need not be the same as each other.
  • nucleotide base As used herein, a “nucleotide base, ” “nucleotide, ” or “nucleobase” is a heterocyclic pyrimidine or purine compound, which is a standard constituent of all nucleic acids, and includes the bases that form the nucleotides adenine, guanine, cytosine, thymine, and uracil.
  • a nucleobase may further be modified to include, though not intended to be limiting: universal bases, hydrophobic bases, promiscuous bases, size-expanded bases, and fluorinated bases.
  • ribonucleotide or “nucleotide” may be used herein to refer to an unmodified nucleotide, a modified nucleotide, or a surrogate replacement moiety.
  • guanine, cytosine, adenine, and uracil can be replaced by other moieties without substantially altering the base pairing properties of an oligonucleotide comprising a nucleotide bearing such replacement moiety.
  • modified RNAs contemplated for use in methods and compositions described herein are peptide nucleic acids (PNAs) that have the ability to form the required duplex structure and that permit or mediate the specific degradation of a target RNA via a RISC pathway.
  • PNAs peptide nucleic acids
  • a CFB RNA interference agent includes a single stranded RNA that interacts with a target CFB RNA sequence to direct the cleavage of the target CFB RNA.
  • Modified RNA backbones can include, for example, phosphorothioates, chiral phosphorothioates, phosphorodithioates, phosphotriesters, aminoalkylphosphotriesters, methyl and other alkyl phosphonates including 3'-alkylene phosphonates and chiral phosphonates, phosphinates, phosphoramidates including 3'-amino phosphoramidate and aminoalkylphosphoramidates, thionophosphoramidates, thionoalkylphosphonates, thionoalkylphosphotriesters, and boranophosphates having normal 3'-5' linkages, 2'-5' linked analogs of these, and those) having inverted polarity wherein the adjacent pairs of nucleoside units are linked 3'-5' to 5'-3' or 2'-5' to 5'-2'.
  • Modified RNA backbones that do not include a phosphorus atom therein have backbones that are formed by short chain alkyl or cycloalkyl internucleoside linkages, mixed heteroatoms and alkyl or cycloalkyl internucleoside linkages, or one or more short chain heteroatomic or heterocyclic internucleoside linkages.
  • morpholino linkages formed in part from the sugar portion of a nucleoside
  • siloxane backbones siloxane backbones
  • sulfide, sulfoxide and sulfone backbones formacetyl and thioformacetyl backbones
  • methylene formacetyl and thioformacetyl backbones alkene containing backbones
  • sulfamate backbones methyleneimino and methylenehydrazino backbones
  • sulfonate and sulfonamide backbones amide backbones
  • others having mixed N, O, S and CH 2 component parts.
  • Means of preparing modified RNA backbones that do not include a phosphorus atom are routinely practiced in the art and such methods can be used to prepare certain modified CFB dsRNA agents, certain modified CFB antisense polynucleotides, and/or certain modified CFB sense polynucleotides of the invention.
  • RNA mimetics are included in CFB dsRNAs, CFB antisense polynucleotides, and/or CFB sense polynucleotides, such as, but not limited to: replacement of the sugar and the internucleoside linkage, i.e., the backbone, of the nucleotide units with novel groups.
  • base units are maintained for hybridization with an appropriate CFB nucleic acid target compound.
  • a peptide nucleic acid (PNA) is referred to as a peptide nucleic acid (PNA) .
  • RNA In PNA compounds, the sugar backbone of an RNA is replaced with an amide containing backbone, in particular an aminoethylglycine backbone.
  • the nucleobases are retained and are bound directly or indirectly to aza nitrogen atoms of the amide portion of the backbone.
  • Means of preparing RNA mimetics are routinely practiced in the art and such methods can be used to prepare certain modified CFB dsRNA agents of the invention.
  • RNAs with phosphorothioate backbones and oligonucleosides with heteroatom backbones and in particular -CH 2 -NH-CH 2 -, -CH 2 -N (CH 3 ) -O-CH 2 - [known as a methylene (methylimino) or MMI backbone] , -CH 2 -O-N (CH 3 ) -CH 2 -, -CH 2 -N (CH 3 ) -N (CH 3 ) -CH 2 -and -N (CH 3 ) -CH 2 - [wherein the native phosphodiester backbone is represented as -O-P-O-CH 2 -] .
  • RNAs with phosphorothioate backbones and oligonucleosides with heteroatom backbones are routinely practiced in the art and such methods can be used to prepare certain modified CFB dsRNA agents, certain CFB antisense polynucleotides, and/or certain CFB sense polynucleotides of the invention.
  • Modified RNAs can also contain one or more substituted sugar moieties.
  • CFB dsRNAs, CFB antisense polynucleotides, and/or CFB sense polynucleotides of the invention may comprise one of the following at the 2' position: OH; F; O-, S-, or N-alkyl; O-, S-, or N-alkenyl; O-, S-or N-alkynyl; or O-alkyl-O-alkyl, wherein the alkyl, alkenyl and alkynyl may be substituted or unsubstituted C 1 to C 10 alkyl or C 2 to C 10 alkenyl and alkynyl.
  • Exemplary suitable modifications include O [ (CH 2 ) n O] m CH 3 , O (CH 2 ) n OCH 3 , O (CH 2 ) n NH 2 , O (CH 2 ) n CH 3 , O (CH 2 ) n ONH 2 , and O (CH 2 ) n ON [ (CH 2 ) n CH 3 ) ] 2 , where n and m are from 1 to about 10.
  • dsRNAs include one of the following at the 2' position: C 1 to C 10 lower alkyl, substituted lower alkyl, alkaryl, aralkyl, O-alkaryl or O-aralkyl, SH, SCH 3 , OCN, Cl, Br, CN, CF 3 , OCF 3 , SOCH 3 , SO 2 CH 3 , ONO 2 , NO 2 , N 3 , NH 2 , heterocycloalkyl, heterocycloalkaryl, aminoalkylamino, polyalkylamino, substituted silyl, an RNA cleaving group, a reporter group, an intercalator, a group for improving the pharmacokinetic properties of a CFB dsRNA agent, or a group for improving the pharmacodynamic properties of a CFB dsRNA agent, CFB antisense polynucleotide, and/or CFB sense polynucleotide, and other substituents having similar properties.
  • the modification includes a 2'-methoxyethoxy (2'-O-CH 2 CH 2 OCH 3 , also known as 2'-O- (2-methoxyethyl) or 2'-MOE) (Martin et al., Helv. Chim. Acta, 1995, 78: 486-504) i.e., an alkoxy-alkoxy group.
  • Another exemplary modification is 2'-dimethylaminooxyethoxy, i.e., a O (CH 2 ) 2 ON (CH 3 ) 2 group, also known as 2'-DMAOE, as described in examples herein below, and 2'-dimethylaminoethoxyethoxy (also known in the art as 2'-O-dimethylaminoethoxyethyl or 2'-DMAEOE) , i.e., 2'-O-CH 2 -O-CH 2 -N (CH 2 ) 2 .
  • Means of preparing modified RNAs such as those described are routinely practiced in the art and such methods can be used to prepare certain modified CFB dsRNA agents of the invention.
  • modifications include 2'-methoxy (2'-OCH 3 ) , 2'-aminopropoxy (2'-OCH 2 CH 2 CH 2 NH 2 ) and 2'-fluoro (2'-F) .
  • Similar modifications can also be made at other positions on the RNA of a CFB dsRNA agent, CFB antisense polynucleotide, and/or CFB sense polynucleotide of the invention, particularly the 3' position of the sugar on the 3' terminal nucleotide or in 2'-5' linked CFB dsRNAs, CFB antisense polynucleotides, or CFB sense polynucleotides, and the 5' position of 5' terminal nucleotide.
  • CFB dsRNA agents, CFB antisense polynucleotides, and/or CFB sense polynucleotides may also have sugar mimetics such as cyclobutyl moieties in place of the pentofuranosyl sugar.
  • sugar mimetics such as cyclobutyl moieties in place of the pentofuranosyl sugar.
  • An CFB dsRNA agent, CFB antisense polynucleotide, and/or CFB sense polynucleotide may, in some embodiments, include nucleobase (often referred to in the art simply as "base” ) modifications or substitutions.
  • nucleobase often referred to in the art simply as "base”
  • “unmodified” or “natural” nucleobases include the purine bases adenine and guanine, and the pyrimidine bases thymine, cytosine and uracil.
  • Modified nucleobases include other synthetic and natural nucleobases such as 5-methylcytosine (5-Me-C) , 5-hydroxymethyl cytosine, xanthine, hypoxanthine, 2-aminoadenine, 6-methyl and other alkyl derivatives of adenine and guanine, 2-propyl and other alkyl derivatives of adenine and guanine, 2-thiouracil, 2-thiothymine and 2-thiocytosine, 5-halouracil and cytosine, 5-propynyl uracil and cytosine, 6-azo uracil, cytosine and thymine, 5-uracil (pseudouracil) , 4-thiouracil, 8-halo, 8-amino, 8-thiol, 8-thioalkyl, 8-hydroxyl anal other 8-substituted adenines and guanines, 5-halo, particularly 5-bromo, 5-trifluoromethyl
  • CFB dsRNA agents of the invention are known in the art, see for example: Modified Nucleosides in Biochemistry, Biotechnology and Medicine, Herdewijn, P. Ed. Wiley-VCH, 2008; The Concise Encyclopedia Of Polymer Science And Engineering, pages 858-859, Kroschwitz, J.L, Ed. John Wiley & Sons, 1990, English et al., Angewandte Chemie, International Edition, 1991, 30, 613, Sanghvi, Y S., Chapter 15, dsRNA Research and Applications, pages 289-302, Crooke, S.T. and Lebleu, B., Ed., CRC Press, 1993.
  • Means of preparing dsRNAs, CFB antisense strand polynucleotides and/or CFB sense strand polynucleotides that comprise nucleobase modifications and/or substitutions such as those described herein are routinely practiced in the art and such methods can be used to prepare certain modified CFB dsRNA agents, CFB sense polynucleotides, and/or CFB antisense polynucleotides of the invention.
  • CFB dsRNA agents, CFB antisense polynucleotides, and/or CFB sense polynucleotides of the invention include RNA modified to include one or more locked nucleic acids (LNA) .
  • LNA locked nucleic acids
  • a locked nucleic acid is a nucleotide with a modified ribose moiety comprising an extra bridge connecting the 2' and 4' carbons. This structure effectively “locks” the ribose in the 3'-endo structural conformation.
  • the addition of locked nucleic acids in a CFB dsRNA agent, CFB antisense polynucleotides, and/or CFB sense polynucleotides of the invention may increase stability in serum, and to reduce off-target effects (Elmen, J.
  • CFB dsRNA compounds, sense polynucleotides, and/or antisense polynucleotides of the invention include at least one modified nucleotide, wherein the at least one modified nucleotide comprises: a 2’-O-methyl nucleotide, 2’-Fluoro nucleotide, 2’-deoxy nucleotide, 2’ 3’-seco nucleotide mimic, locked nucleotide, 2’-F-Arabino nucleotide, 2’-methoyxyethyl nucleotide, 2’-amino-modified nucleotide, 2’-alkyl-modified nucleotide, mopholino nucleotide, and 3’-OMe nucleotide, a nucleotide comprising a 5’-phosphorothioate group, a nucleotide comprising vinyl phosphonate, a nucleotide comprising
  • CFB dsRNA compounds 3’ and 5’ end of sense polynucleotides, and/or 3’ end of antisense polynucleotides of the invention, include at least one modified nucleotide, wherein the at least one modified nucleotide comprises: abasic nucleotide, ribitol, inverted nucleotide, inverted abasic nucleotide, inverted 2’-OMe nucleotide, inverted 2’-deoxy nucleotide. It is known to skilled in art, including an abasic or inverted abasic nucleotide at the end of oligonucleotide enhances stability (Czauderna et al.
  • a CFB dsRNA compound includes one or more inverted abasic residues (invab) at either 3’-end or 5’-end, or both 3’-end and 5’-end.
  • invab inverted abasic residues
  • Exemplified inverted abasic residues (invab) include, but are not limited to the following:
  • CFB dsRNA compounds 3’ and 5’ end of sense polynucleotides, and/or 3’ end of antisense polynucleotides of the invention, include at least one modified nucleotide, wherein the at least one modified nucleotide comprises: isomannide residues or stereoisomer of said isomannide residues.
  • isomannide residues or stereoisomers of said isomannide residues include, but are not limited to:
  • isomannide residues include, but are not limited to, the following:
  • the isomannide nucleotides may further conjugate to one or more targeting groups or delivery molecules, such as GalNAc moieties.
  • CFB dsRNA compounds, antisense polynucleotides of the invention include at least one modified nucleotide, wherein the at least one modified nucleotide comprises unlocked nucleic acid nucleotide (UNA) or/and glycol nucleic acid nucleotide (GNA) .
  • UNA and GNA are thermally destabilizing chemical modifications, can significantly improves the off-target profile of a siRNA compound (Janas, et al., Selection of GalNAc-conjugated siRNAs with limited off-target-driven rat hepatotoxicity. Nat Commun. 2018; 9 (1) : 723.
  • Another modification that may be included in the RNA of certain embodiments of CFB dsRNA agents, CFB antisense polynucleotides, and/or CFB sense polynucleotides of the invention comprises chemically linking to the RNA one or more ligands, moieties or conjugates that enhance one or more characteristics of the CFB dsRNA agent, CFB antisense polynucleotide, and/or CFB sense polynucleotide, respectively.
  • Non-limiting examples of characteristics that may be enhanced are: CFB dsRNA agent, CFB antisense polynucleotide, and/or CFB sense polynucleotide activity, cellular distribution, delivery of a CFB dsRNA agent, pharmacokinetic properties of a CFB dsRNA agent, and cellular uptake of the CFB dsRNA agent.
  • a CFB dsRNA agent comprises one or more targeting groups or linking groups, which in certain embodiments of CFB dsRNA agents of the invention are conjugated to the sense strand.
  • a non-limiting example of a targeting group is a compound comprising N-acetyl-galactosamine (GalNAc) .
  • a CFB dsRNA agent comprises a targeting compound that is conjugated to the 5'-terminal end of the sense strand.
  • a CFB dsRNA agent comprises a targeting compound that is conjugated to the 3'-terminal end of the sense strand.
  • a CFB dsRNA agent comprises a targeting group that comprises GalNAc.
  • a CFB dsRNA agent does not include a targeting compound conjugated to one or both of the 3'-terminal end and the 5'-terminal end of the sense strand. In certain embodiments of the invention a CFB dsRNA agent does not include a GalNAc containing targeting compound conjugated to one or both of the 5'-terminal end and the 3'-terminal end of the sense strand.
  • targeting and linking agents include but are not limited to lipid moieties such as a cholesterol moiety (Letsinger et al., Proc. Natl. Acid. Sci. USA, 1989, 86: 6553-6556) , cholic acid (Manoharan et al., Biorg. Med. Chem. Let., 1994, 4: 1053-1060) , a thioether, e.g., beryl-S-tritylthiol (Manoharan et al., Ann. N.Y. Acad. Sci., 1992, 660: 306-309; Manoharan et al., Biorg.
  • lipid moieties such as a cholesterol moiety (Letsinger et al., Proc. Natl. Acid. Sci. USA, 1989, 86: 6553-6556) , cholic acid (Manoharan et al., Biorg. Med. Chem. Let., 1994, 4: 1053-1060)
  • Acids Res., 1990, 18: 3777-3783) a polyamine or a polyethylene glycol chain (Manoharan et al., Nucleosides & Nucleotides, 1995, 14: 969-973) , or adamantane acetic acid (Manoharan et al., Tetrahedron Lett., 1995, 36: 3651-3654) , a palmityl moiety (Mishra et al., Biochim. Biophys. Acta, 1995, 1264: 229-237) , or an octadecylamine or hexylamino-carbonyloxycholesterol moiety (Crooke et al., J. Pharmacol. Exp. Ther., 1996, 277: 923-937) .
  • compositions comprising a CFB dsRNA agent, CFB antisense polynucleotide, and/or CFB sense polynucleotide may comprise a ligand that alters distribution, targeting, or etc. of the CFB dsRNA agent.
  • the ligand increases affinity for a selected target, e.g., molecule, cell or cell type, compartment, e.g., a cellular or organ compartment, tissue, organ or region of the body, as, e.g., compared to a species absent such a ligand.
  • a ligand useful in a composition and/or method of the invention may be a naturally occurring substance, such as a protein (e.g., human serum albumin (HSA) , low-density lipoprotein (LDL) , or globulin) ; a carbohydrate (e.g., a dextran, pullulan, chitin, chitosan, inulin, cyclodextrin or hyaluronic acid) ; or a lipid.
  • a ligand may also be a recombinant or synthetic molecule, such as a synthetic polymer, e.g., a synthetic polyamino acid or polyamine.
  • polyamino acids examples include a polylysine (PLL) , poly L-aspartic acid, poly L-glutamic acid, styrene-maleic acid anhydride copolymer, poly (L-lactide-co-glycolied) copolymer, divinyl ether-maleic anhydride copolymer, N- (2-hydroxypropyl) methacrylamide copolymer (HMPA) , polyethylene glycol (PEG) , polyvinyl alcohol (PVA) , polyurethane, poly (2-ethylacryllic acid) , N-isopropylacrylamide polymers, or polyphosphazine.
  • PLL polylysine
  • poly L-aspartic acid poly L-glutamic acid
  • styrene-maleic acid anhydride copolymer poly (L-lactide-co-glycolied) copolymer
  • divinyl ether-maleic anhydride copolymer N-
  • polyamines include: polyethylenimine, polylysine (PLL) , spermine, spermidine, polyamine, pseudopeptide-polyamine, peptidomimetic polyamine, dendrimer polyamine, arginine, amidine, protamine, cationic lipid, cationic porphyrin, quaternary salt of a polyamine, or an alpha helical peptide.
  • a ligand included in a composition and/or method of the invention may comprise a targeting group, non-limiting examples of which are a cell or tissue targeting agent, e.g., a lectin, glycoprotein, lipid or protein, e.g., an antibody that binds to a specified cell type such as a kidney cell or a liver cell.
  • a targeting group non-limiting examples of which are a cell or tissue targeting agent, e.g., a lectin, glycoprotein, lipid or protein, e.g., an antibody that binds to a specified cell type such as a kidney cell or a liver cell.
  • a targeting group can be a thyrotropin, melanotropin, lectin, glycoprotein, surfactant protein A, Mucin carbohydrate, multivalent lactose, multivalent galactose, N-acetyl-galactosamine, N-acetyl-gulucosamine multivalent mannose, multivalent fucose, glycosylated polyaminoacids, multivalent galactose, transferrin, bisphosphonate, polyglutamate, polyaspartate, a lipid, cholesterol, a steroid, bile acid, folate, vitamin B12, vitamin A, biotin, or an RGD peptide or RGD peptide mimetic.
  • ligands include dyes, intercalating agents (e.g. acridines) , cross-linkers (e.g. psoralene, mitomycin C) , porphyrins (TPPC4, texaphyrin, Sapphyrin) , polycyclic aromatic hydrocarbons (e.g., phenazine, dihydrophenazine) , artificial endonucleases (e.g.
  • EDTA lipophilic molecules, e.g., cholesterol, cholic acid, adamantane acetic acid, 1-pyrene butyric acid, dihydrotestosterone, 1, 3-Bis-O (hexadecyl) glycerol, geranyloxyhexyl group, hexadecylglycerol, borneol, menthol, 1, 3-propanediol, heptadecyl group, palmitic acid, myristic acid, O3- (oleoyl) lithocholic acid, O3- (oleoyl) cholenic acid, dimethoxytrityl, or phenoxazine) and peptide conjugates (e.g., antennapedia peptide, Tat peptide) , alkylating agents, phosphate, amino, mercapto, PEG (e.g., PEG-40K) , MPEG, [MPEG] 2 , polyamin
  • a ligand included in a composition and/or method of the invention may be a protein, e.g., glycoprotein, or peptide, for example a molecule with a specific affinity for a co-ligand, or an antibody, for example an antibody, that binds to a specified cell type such as a cancer cell, endothelial cell, cardiac cell, or bone cell.
  • a ligand useful in an embodiment of a composition and/or method of the invention can be a hormone or hormone receptor.
  • a ligand useful in an embodiment of a composition and/or method of the invention can be a lipid, lectin, carbohydrates, vitamin, cofactos, multivalent lactose, multivalent galactose, N-acetyl-galactosamine, N-acetyl-gulucosamine multivalent mannose, or multivalent fucose.
  • a ligand useful in an embodiment of a composition and/or method of the invention can be a substance that can increase uptake of the CFB dsRNA agent into the cell, for example, by disrupting the cell's cytoskeleton, e.g., by disrupting the cell's microtubules, microfilaments, and/or intermediate filaments.
  • Non-limiting examples of this type of agent are: taxon, vincristine, vinblastine, cytochalasin, nocodazole, japlakinolide, latrunculin A, phalloidin, swinholide A, indanocine, and myoservin.
  • a ligand attached to a CFB dsRNA agent of the invention functions as a pharmacokinetic (PK) modulator.
  • PK modulator that may be used in compositions and methods of the invention includes but is not limited to: a lipophiles, a bile acid, a steroid, a phospholipid analogue, a peptide, a protein binding agent, PEG, a vitamin, cholesterol, a fatty acid, cholic acid, lithocholic acid, dialkylglycerides, diacylglyceride, a phospholipid, a sphingolipid, naproxen, ibuprofen, vitamin E, biotin, an aptamer that binds a serum protein, etc.
  • Oligonucleotides comprising a number of phosphorothioate linkages are also known to bind to serum protein, thus short oligonucleotides, e.g., oligonucleotides of about 5 bases, 10 bases, 15 bases or 20 bases, comprising multiple of phosphorothioate linkages in the backbone may also be used in compositions and/or methods of the invention as ligands.
  • a CFB dsRNA agent is in a composition.
  • a composition of the invention may include one or more CFB dsRNA agent and optionally one or more of a pharmaceutically acceptable carrier, a delivery agent, a targeting agent, detectable label, etc.
  • a non-limiting example of a targeting agent that may be useful according to some embodiments of methods of the invention is an agent that directs a CFB dsRNA agent of the invention to and/or into a cell to be treated.
  • a targeting agent of choice will depend upon such elements as: the nature of the CFB-associated disease or condition, and on the cell type being targeted.
  • a therapeutic agent comprises a CFB dsRNA agent with only a delivery agent, such as a delivery agent comprising N-Acetylgalactosamine (GalNAc) , without any additional attached elements.
  • a CFB dsRNA agent may be attached to a delivery compound comprising GalNAc and included in a composition comprising a pharmaceutically acceptable carrier and administered to a cell or subject without any detectable labels, or targeting agents, etc. attached to the CFB dsRNA agent.
  • Labeling agents may be used in certain methods of the invention to determine the location of a CFB dsRNA agent in cells and tissues and may be used to determine a cell, tissue, or organ location of a treatment composition comprising a CFB dsRNA agent that has been administered in methods of the invention.
  • Procedures for attaching and utilizing labeling agents such as enzymatic labels, dyes, radiolabels, etc. are well known in the art.
  • a labeling agent is attached to one or both of a sense polynucleotide and an antisense polynucleotide included in a CFB dsRNA agent.
  • Certain embodiments of methods of the invention includes delivery of a CFB dsRNA agent into a cell.
  • delivery means facilitating or effecting uptake or absorption into the cell. Absorption or uptake of a CFB dsRNA agent can occur through unaided diffusive or active cellular processes, or by use of delivery agents, targeting agents, etc. that may be associated with a CFB dsRNA agent of the invention.
  • Delivery means that are suitable for use in methods of the invention include, but are not limited to: in vivo delivery, in which a CFB dsRNA agent is in injected into a tissue site or administered systemically. In some embodiments of the invention, a CFB dsRNA agent is attached to a delivery agent.
  • Non-limiting examples of methods that can be used to deliver CFB dsRNA agents to cells, tissues and/or subjects include: CFB dsRNA-GalNAc conjugates, SAMiRNA technology, LNP-based delivery methods, and naked RNA delivery. These and other delivery methods have been used successfully in the art to deliver therapeutic RNAi agents for treatment of various diseases and conditions, such as but not limited to: liver diseases, acute intermittent porphyria (AIP) , hemophilia, pulmonary fibrosis, etc. Details of various delivery means are found in publications such as: Nikam, R.R. & K.R. Gore (2016) Nucleic Acid Ther, 28 (4) , 209-224 Aug 2018; Springer A.D. & S.F.
  • LNPs lipid nanoparticles
  • CFB dsRNA agent of the invention Some embodiments of the invention comprise use of lipid nanoparticles (LNPs) to deliver a CFB dsRNA agent of the invention to a cell, tissue, and/or subject.
  • LNPs are routinely used for in vivo delivery of CFB dsRNA agents, including therapeutic CFB dsRNA agents.
  • One benefit of using an LNP or other delivery agent is an increased stability of the CFB RNA agent when it is delivered to a subject using the LNP or other delivery agent.
  • an LNP comprises a cationic LNP that is loaded with one or more CFB RNAi molecules of the invention.
  • the LNP comprising the CFB RNAi molecule (s) is administered to a subject, the LNPs and their attached CFB RNAi molecules are taken up by cells via endocytosis, their presence results in release of RNAi trigger molecules, which mediate RNAi.
  • a delivery agent that may be used in embodiments of the invention to delivery a CFB dsRNA agent of the invention to a cell, tissue and/or subject is an agent comprising GalNAc that is attached to a CFB dsRNA agent of the invention and delivers the CFB dsRNA agent to a cell, tissue, and/or subject.
  • agents comprising GalNAc that can be used in certain embodiments of methods and composition of the invention are disclosed in PCT Application: WO2020191183A1 (incorporated herein in its entirety) .
  • a non-limiting example of a GalNAc targeting ligand that can be used in compositions and methods of the invention to deliver a CFB dsRNA agent to a cell is a targeting ligand cluster.
  • GalNAc Ligand with phosphodiester link GLO
  • GalNAc Ligand with phosphorothioate link GLO
  • GLX-n GalNAc Ligand with phosphorothioate link
  • the term “GLX-n” may be used herein to indicate the attached GalNAc-containing compound is any one of compounds GLS-1*, GLS-2*, GLS-3*, GLS-4*, GLS-5*, GLS-6*, GLS-7*, GLS-8*, GLS-9*, GLS-10*, GLS-11*, GLS-12*, GLS-13*, GLS-14*, GLS-15*, GLS-16*, GLO-1, GLO-2, GLO-3, GLO-4, GLO-5, GLO-6, GLO-7, GLO-8, GLO-9, GLO-10, GLO-11, GLO-12, GLO-13, GLO-14, GLO-15, and GLO-16, the structure of each of which is shown below, with the below with location of attachment of the GalNAc-targeting lig
  • any RNAi and dsRNA molecule of the invention can be attached to the GLS-1*, GLS-2*, GLS-3*, GLS-4*, GLS-5*, GLS-6*, GLS-7*, GLS-8*, GLS-9*, GLS-10*, GLS-11*, GLS-12*, GLS-13*, GLS-14*, GLS-15*, GLS-16*, GLO-1, GLO-2, GLO-3, GLO-4, GLO-5, GLO-6, GLO-7, GLO-8, GLO-9, GLO-10, GLO-11, GLO-12, GLO-13, GLO-14, GLO-15, and GLO-16, GLO-1 through GLO-16 and GLS-1*through GLS-16*structures are shown below.
  • the aforesaid isomannide nucleotides may further conjugate to one or more GalNAc targeting ligands.
  • Specific examples of isomannide nucleotides conjugated to a GalNAc targeting ligand include, but are not limited to:
  • olig each independently represents a polynucleotide moiety.
  • in vivo delivery can also be by a beta-glucan delivery system, such as those described in U.S. Pat. Nos. 5,032,401 and 5,607,677, and U.S. Publication No. 2005/0281781, which are hereby incorporated by reference in their entirety.
  • a CFB dsRNA is delivered without a targeting agent. These RNAs may be delivered as “naked” RNA molecules.
  • a CFB dsRNA of the invention may be administered to a subject to treat a CFB-associated disease or condition in the subject, such as a cardiovascular disease, in a pharmaceutical composition comprising the RNAi agent, but not including a targeting agent such as a GalNAc targeting compound.
  • RNAi delivery means such as but not limited to those described herein and those used in the art, can be used in conjunction with embodiments of CFB RNAi agents and treatment methods described herein.
  • CFB dsRNA agents of the invention may be administered to a subject in an amount and manner effective to reduce a level and activity of CFB polypeptide in a cell and/or subject.
  • one or more CFB dsRNA agents are administered to a cell and/or subject to treat a disease or condition associated with CFB expression and activity.
  • Methods of the invention include administering one or more CFB dsRNA agents to a subject in need of such treatment to reduce a disease or condition associated with CFB expression in the subject.
  • CFB dsRNA agents or CFB antisense polynucleotide agents of the invention can be administered to reduce CFB expression and/or activity in one more of in vitro, ex vivo, and in vivo cells.
  • a level, and thus an activity, of CFB polypeptide in a cell is reduced by delivering (e.g. introducing) an CFB dsRNA agent or CFB antisense polynucleotide agent into a cell.
  • Targeting agents and methods may be used to aid in delivery of a CFB dsRNA agent or CFB antisense polynucleotide agent to a specific cell type, cell subtype, organ, spatial region within a subject, and/or to a sub-cellular region within a cell.
  • An CFB dsRNA agent can be administered in certain methods of the invention singly or in combination with one or more additional CFB dsRNA agents. In some embodiments, 2, 3, 4, or more independently selected CFB dsRNA agents are administered to a subject.
  • an CFB dsRNA agent is administered to a subject to treat a CFB-associated disease or condition in conjunction with one or more additional therapeutic regimens for treating the CFB-associate disease or condition.
  • additional therapeutic regimens are: administering one or more CFB antisense polynucleotides of the invention, administering a non-CFB dsRNA therapeutic agent, and a behavioral modification.
  • An additional therapeutic regimen may be administered at a time that is one or more of: prior to, simultaneous with, and following administration of a CFB dsRNA agent of the invention.
  • Non-limiting examples of non-CFB dsRNA therapeutic agents are: an inhibitor of C5, such as an anticomplement component C5 antibody, or antigen-binding fragment thereof (e.g., eculizumab, ravulizumab-cwvz, or Polimab (REGN3918) ) or a C5 peptide inhibitor (e.g., zilucoplan) .
  • an inhibitor of C5 such as an anticomplement component C5 antibody, or antigen-binding fragment thereof (e.g., eculizumab, ravulizumab-cwvz, or Polimab (REGN3918)
  • a C5 peptide inhibitor e.g., zilucoplan
  • Eculizumab is a humanized monoclonal IgG2/4, kappa light chain antibody that specifically binds complement component C5 with high affinity and inhibits cleavage of C5 to C5a and C5b, thereby inhibiting the generation of the terminal complement complex C5b-9.
  • Ravulizumab-cwvz is a humanized IgG2/4 monoclonal antibody that specifically binds complement component C5 with high affinity and inhibits cleavage of C5 to C5a and C5b, thereby inhibiting the generation of the terminal complement complex C5b-9.
  • Pozelimab (also known as H4H12166P, described in US20170355757) is a fully-human IgG4 monoclonal antibody designed to block complement factor C5.
  • Zilucoplan is a synthetic, macrocyclic peptide that binds complement component 5 (C5) with sub-nanomolar affinity and allosterically inhibits its cleavage into C5a and C5b upon activation of the classical, alternative, or lectin pathways.
  • the additional therapeutic is a C3 peptide inhibitor, or analog thereof.
  • the C3 peptide inhibitor is compstatin.
  • Compstatin is a cyclic tridecapeptide with potent and selective C3 inhibitory activity.
  • CFB-associated disease or condition in a subject and may be administered to a subject in combination with the administration of one or more CFB dsRNA agents of the invention to treat the CFB-associated disease or condition.
  • a CFB dsRNA agent of the invention administered to a cell or subject to treat a CFB-associated disease or condition may act in a synergistic manner with one or more other therapeutic agents or activities and increase the effectiveness of the one or more therapeutic agents or activities and/or to increase the effectiveness of the CFB dsRNA agent at treating the CFB-associated disease or condition.
  • Treatment methods of the invention that include administration of a CFB dsRNA agent can be used prior to the onset of a CFB-associated disease or condition and/or when a CFB-associated disease or condition is present, including at an early stage, mid-stage, and late stage of the disease or condition and all times before and after any of these stages.
  • Methods of the invention may also be to treat subjects who have previously been treated for a CFB-associated disease or condition with one or more other therapeutic agents and/or therapeutic activities that were not successful, were minimally successful, and/or are no longer successful at treating the CFB-associated disease or condition in the subject.
  • a CFB dsRNA agent can be delivered into a cell using a vector.
  • CFB dsRNA agent transcription units can be included in a DNA or RNA vector.
  • Vectors can be used in methods of the invention that result in transient expression of CFB dsRNA, for example for at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more hours, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more weeks.
  • the length of the transient expression can be determined using routine methods based on elements such as, but not limited to the specific vector construct selected and the target cell and/or tissue.
  • transgenes can be introduced as a linear construct, a circular plasmid, or a viral vector, which can be an integrating or non-integrating vector.
  • the transgene can also be constructed to permit it to be inherited as an extrachromosomal plasmid (Gassmann, et al., Proc. Natl. Acad. Sci. USA (1995) 92: 1292) .
  • An individual strand or strands of a CFB dsRNA agent can be transcribed from a promoter on an expression vector. Where two separate strands are to be expressed to generate, for example, a dsRNA, two separate expression vectors can be co-introduced to a cell using means such as transfection or infection.
  • each individual strand of a CFB dsRNA agent of the invention can be transcribed by promoters that are both included on the same expression vector.
  • a CFB dsRNA agent is expressed as inverted repeat polynucleotides joined by a linker polynucleotide sequence such that the CFB dsRNA agent has a stem and loop structure.
  • RNA expression vectors are DNA plasmids or viral vectors.
  • Expression vectors useful in embodiments of the invention can be compatible with eukaryotic cells.
  • Eukaryotic cell expression vectors are routinely used in the art and are available from a number of commercial sources. Delivery of CFB dsRNA expressing vectors can be systemic, such as by intravenous or intramuscular administration, by administration to target cells ex-planted from a subject followed by reintroduction into the subject, or by any other means that allows for introduction into a desired target cell.
  • Viral vector systems that may be included in an embodiment of a method of the include, but are not limited to, (a) adenovirus vectors; (b) retrovirus vectors, including but not limited to lentiviral vectors, moloney murine leukemia virus, etc.; (c) adeno-associated virus vectors; (d) herpes simplex virus vectors; (e) SV 40 vectors; (f) polyoma virus vectors; (g) papilloma virus vectors; (h) picornavirus vectors; (i) pox virus vectors such as an orthopox, e.g., vaccinia virus vectors or avipox, e.g.
  • pox virus vectors such as an orthopox, e.g., vaccinia virus vectors or avipox, e.g.
  • Constructs for the recombinant expression of a CFB dsRNA agent may include regulatory elements, such as promoters, enhancers, etc., which may be selected to provide constitutive or regulated/inducible expression.
  • regulatory elements such as promoters, enhancers, etc.
  • Viral vector systems, and the use of promoters and enhancers, etc. are routine in the art and can be used in conjunction with methods and compositions described herein.
  • Certain embodiments of the invention include use of viral vectors for delivery of CFB dsRNA agents into cells.
  • Numerous adenovirus-based delivery systems are routinely used in the art for deliver to, for example, lung, liver, the central nervous system, endothelial cells, and muscle.
  • Non-limiting examples of viral vectors that may be used in methods of the invention are: AAV vectors, a pox virus such as a vaccinia virus, a Modified Virus Ankara (MVA) , NYVAC, an avipox such as fowl pox or canary pox.
  • Certain embodiments of the invention include methods of delivering CFB dsRNA agents into cells using a vector and such vectors may be in a pharmaceutically acceptable carrier that may, but need not, include a slow release matrix in which the gene delivery vehicle is imbedded.
  • a vector for delivering a CFB dsRNA can be produced from a recombinant cell, and a pharmaceutical composition of the invention may include one or more cells that produced the CFB dsRNA delivery system.
  • compositions Containing CFB dsRNA or ssRNA agents Containing CFB dsRNA or ssRNA agents
  • Certain embodiments of the invention include use of pharmaceutical compositions containing a CFB dsRNA agent or CFB antisense polynucleotide agent and a pharmaceutically acceptable carrier.
  • the pharmaceutical composition containing the CFB dsRNA agent or CFB antisense polynucleotide agent can be used in methods of the invention to reduce CFB gene expression and CFB activity in a cell and is useful to treat a CFB-associated disease or condition.
  • Such pharmaceutical compositions can be formulated based on the mode of delivery.
  • Non-limiting examples of formulations for modes of delivery are: a composition formulated for subcutaneous delivery, a composition formulated for systemic administration via parenteral delivery, a composition formulated for intravenous (IV) delivery, a composition formulated for intrathecal delivery, a composition formulated for direct delivery into brain, etc.
  • Administration of a pharmaceutic composition of the invention to deliver a CFB dsRNA agent or CFB antisense polynucleotide agent into a cell may be done using one or more means such as: topical (e.g., by a transdermal patch) , pulmonary, e.g., by inhalation or insufflation of powders or aerosols, including by nebulizer; intratracheal, intranasal, epidermal and transdermal, oral or parenteral.
  • topical e.g., by a transdermal patch
  • pulmonary e.g., by inhalation or insufflation of powders or aerosols, including by nebulizer
  • intratracheal intranasal, epidermal and transdermal, oral or parenteral.
  • Parenteral administration includes intravenous, intraarterial, subcutaneous, intraperitoneal or intramuscular injection or infusion; subdermal, e.g., via an implanted device; or intracranial, e.g., by intraparenchymal, intrathecal or intraventricular, administration.
  • a CFB dsRNA agent or CFB antisense polynucleotide agent can also be delivered directly to a target tissue, for example directly into the liver, directly into a kidney, etc.
  • delivering a CFB dsRNA agent” or “delivering a CFB antisense polynucleotide agent” into a cell encompasses delivering a CFB dsRNA agent or CFB antisense polynucleotide agent, respectively, directly as well as expressing a CFB dsRNA agent in a cell from an encoding vector that is delivered into a cell, or by any suitable means with which the CFB dsRNA or CFB antisense polynucleotide agent becomes present in a cell.
  • Preparation and use of formulations and means for delivering inhibitory RNAs are well known and routinely used in the art.
  • a “pharmaceutical composition” comprises a pharmacologically effective amount of a CFB dsRNA agent or CFB antisense polynucleotide agent of the invention and a pharmaceutically acceptable carrier.
  • pharmaceutically acceptable carrier refers to a carrier for administration of a therapeutic agent. Such carriers include, but are not limited to, saline, buffered saline, dextrose, water, glycerol, ethanol, and combinations thereof. The term specifically excludes cell culture medium.
  • pharmaceutically acceptable carriers include, but are not limited to pharmaceutically acceptable excipients such as inert diluents, disintegrating agents, binding agents, lubricating agents, sweetening agents, flavoring agents, coloring agents and preservatives.
  • suitable inert diluents include sodium and calcium carbonate, sodium and calcium phosphate, and lactose, while corn starch and alginic acid are suitable disintegrating agents.
  • Binding agents may include starch and gelatin, while the lubricating agent, if present, will generally be magnesium stearate, stearic acid or talc.
  • the tablets may be coated with a material such as glyceryl monostearate or glyceryl distearate, to delay absorption in the gastrointestinal tract. Agents included in drug formulations are described further herein below.
  • pharmacologically effective amount refers to that amount of a CFB dsRNA agent or CFB antisense polynucleotide agent of the invention to produce the intended pharmacological, therapeutic or preventive result.
  • a therapeutically effective amount of a drug for the treatment of that disease or disorder is the amount necessary to effect at least a 10%reduction in that parameter.
  • a therapeutically effective amount of a CFB dsRNA agent or CFB antisense polynucleotide agent can reduce CFB polypeptide levels by at least 10%.
  • Methods of the invention in some aspects comprise contacting a cell with a CFB dsRNA agent or CFB antisense polynucleotide agent in an effective amount to reduce CFB gene expression in the contacted cell.
  • Certain embodiments of methods of the invention comprise administering a CFB dsRNA agent or a CFB antisense polynucleotide agent to a subject in an amount effective to reduce CFB gene expression and treat a CFB-associated disease or condition in the subject.
  • An “effective amount” used in terms of reducing expression of CFB and/or for treating a CFB-associated disease or condition is an amount necessary or sufficient to realize a desired biologic effect.
  • an effective amount of a CFB dsRNA agent or CFB antisense polynucleotide agent to treat a CFB-associated disease or condition could be that amount necessary to (i) slow or halt progression of the disease or condition; or (ii) reverse, reduce, or eliminate one or more symptoms of the disease or condition.
  • an effective amount is that amount of a CFB dsRNA agent or CFB antisense polynucleotide agent that when administered to a subject in need of a treatment of a CFB-associated disease or condition, results in a therapeutic response that prevents and/or treats the disease or condition.
  • an effective amount is that amount of a CFB dsRNA agent or CFB antisense polynucleotide agent of the invention that when combined or co-administered with another therapeutic treatment for a CFB-associated disease or condition, results in a therapeutic response that prevents and/or treats the disease or condition.
  • a biologic effect of treating a subject with a CFB dsRNA agent or CFB antisense polynucleotide agent of the invention may be the amelioration and or absolute elimination of symptoms resulting from the CFB-associated disease or condition.
  • a biologic effect is the complete abrogation of the CFB-associated disease or condition, as evidenced for example, by a diagnostic test that indicates the subject is free of the CFB-associated disease or condition.
  • a non-limiting example of a physiological symptom that may be detected includes a reduction in CFB level in liver of a subject following administration of an agent of the invention. Additional art-known means of assessing the status of a CFB-associated disease or condition can be used to determine an effect of an agent and/or methods of the invention on a CFB-associated disease or condition.
  • an effective amount of a CFB dsRNA agent or CFB antisense polynucleotide agent to decrease CFB polypeptide activity to a level to treat a CFB-associated disease or condition will be determined in clinical trials, establishing an effective dose for a test population versus a control population in a blind study. In some embodiments, an effective amount will be that results in a desired response, e.g., an amount that diminishes a CFB-associated disease or condition in cells, tissues, and/or subjects with the disease or condition.
  • an effective amount of a CFB dsRNA agent or CFB antisense polynucleotide agent to treat a CFB-associated disease or condition that can be treated by reducing CFB polypeptide activity may be the amount that when administered decreases the amount of CFB polypeptide activity in the subject to an amount that is less than the amount that would be present in the cell, tissue, and/or subject without the administration of the CFB dsRNA agent or CFB antisense polynucleotide agent.
  • control amount for a subject is a pre-treatment amount for the subject, in other words, a level in a subject before administration of a CFB agent can be a control level for that subject and compared to a level of CFB polypeptide activity and/or CFB gene expression in the subject following siRNA administered to the subject.
  • the desired response may be reducing or eliminating one or more symptoms of the disease or condition in the cell, tissue, and/or subject.
  • the reduction or elimination may be temporary or may be permanent.
  • the status of a CFB-associated disease or condition can be monitored using methods of determining CFB polypeptide activity, CFB gene expression, symptom evaluation, clinical testing, etc.
  • a desired response to treatment of a CFB-associated disease or condition is delaying the onset or even preventing the onset of the disease or condition.
  • An effective amount of a compound that decreases CFB polypeptide activity may also be determined by assessing physiological effects of administration of a CFB dsRNA agent or CFB antisense polynucleotide agent on a cell or subject, such as a decrease of a CFB-associated disease or condition following administration.
  • Assays and/or symptomatic monitoring of a subject can be used to determine efficacy of a CFB dsRNA agent or CFB antisense polynucleotide agent of the invention, which may be administered in a pharmaceutical compound of the invention, and to determine the presence or absence of a response to the treatment.
  • a non-limiting example is that one or more art-known tests of CH50 activity (a measure of total hemolytic complement) , AH50 (a measure the hemolytic activity of the alternate pathway of complement) , lactate dehydrogenase (LDH) (a measure of intravascular hemolysis) , hemoglobin levels; the level of any one or more of C3, C9, C5, C5a, C5b, and soluble C5b-9 complex.
  • AH50 a measure the hemolytic activity of the alternate pathway of complement
  • LDH lactate dehydrogenase
  • Another non-limiting example is that one or more art-known tests of liver function can be used to determine the status of the CFB-associated lipid imbalance in a subject before and after treatment of the subject with a CFB dsRNA agent of the invention.
  • Some embodiments of the invention include methods of determining an efficacy of an dsRNA agent or CFB antisense polynucleotide agent of the invention administered to a subject, to treat a CFB-associated disease or condition by assessing and/or monitoring one or more “physiological characteristics” of the CFB-associated disease or condition in the subject.
  • Non-limiting examples of physiological characteristics of a CFB-associated disease or condition are the CFB mRNA level, the CFB protein level, or CH50 activity (a measure of total hemolytic complement) , AH50 (a measure the hemolytic activity of the alternate pathway of complement) , lactate dehydrogenase (LDH) (a measure of intravascular hemolysis) , hemoglobin levels; the level of any one or more of C3, C9, C5, C5a, C5b, and soluble C5b-9 complex. Standard means of determining such physiological characteristic are known in the art and include, but are not limited to, blood tests, imaging studies, physical examination, etc.
  • the amount of a CFB dsRNA agent or CFB antisense polynucleotide agent administered to a subject can be modified based, at least in part, on such determinations of disease and/or condition status and/or physiological characteristics determined for a subject.
  • the amount of a treatment may be varied for example by increasing or decreasing the amount of a CFB-dsRNA agent or CFB antisense polynucleotide agent, by changing the composition in which the CFB dsRNA agent or CFB antisense polynucleotide agent, respectively, is administered, by changing the route of administration, by changing the dosage timing and so on.
  • the effective amount of a CFB dsRNA agent or CFB antisense polynucleotide agent will vary with the particular condition being treated, the age and physical condition of the subject being treated; the severity of the condition, the duration of the treatment, the nature of the concurrent therapy (if any) , the specific route of administration, and additional factors within the knowledge and expertise of the health practitioner.
  • an effective amount may depend upon the desired level of CFB polypeptide activity and or CFB gene expression that is effective to treat the CFB-associated disease or condition.
  • a skilled artisan can empirically determine an effective amount of a particular CFB dsRNA agent or CFB antisense polynucleotide agent of the invention for use in methods of the invention without necessitating undue experimentation.
  • an effective prophylactic or therapeutic treatment regimen can be planned that is effective to treat the particular subject.
  • an effective amount of a CFB dsRNA agent or CFB antisense polynucleotide agent of the invention can be that amount that when contacted with a cell results in a desired biological effect in the cell.
  • CFB gene silencing may be determined in any cell expressing CFB, either constitutively or by genomic engineering, and by any appropriate assay.
  • CFB gene expression is reduced by at least 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100%by administration of a CFB dsRNA agent of the invention.
  • CFB gene expression is reduced by at between 5%and 10%, 5% and 25%, 10%and 50%, 10%and 75%, 25%and 75%, 25%and 100%, or 50%and 100%by administration of a CFB dsRNA agent of the invention.
  • CFB dsRNA agents and CFB antisense polynucleotide agents are delivered in pharmaceutical compositions in dosages sufficient to inhibit expression of CFB genes.
  • a dose of CFB dsRNA agent or CFB antisense polynucleotide agent is in a range of 0.01 to 200.0 milligrams per kilogram body weight of the recipient per day, generally in the range of 1 to 50 mg per kilogram body weight, 5 to 40 mg/kg body weight, 10 to 30 mg/kg body weight, 1 to 20 mg/kg body weight, 1 to 10 mg/kg body weight, 4 to 15 mg/kg body weight per day, inclusive.
  • the CFB dsRNA agent or CFB antisense polynucleotide agent can be administered in an amount that is from about 0.01 mg/kg, 0.05 mg/kg, 0.1 mg/kg, 0.2 mg/kg, 0.3 mg/kg, 0.4 mg/kg, 0.5 mg/kg, 1 mg/kg, 1.1 mg/kg, 1.2 mg/kg, 1.3 mg/kg, 1.4 mg/kg, 1.5 mg/kg, 1.6 mg/kg, 1.7 mg/kg, 1.8 mg/kg, 1.9 mg/kg, 2 mg/kg, 2.1 mg/kg, 2.2 mg/kg, 2.3 mg/kg, 2.4 mg/kg, 2.5 mg/kg, 2.6 mg/kg, 2.7 mg/kg, 2.8 mg/kg, 2.9 mg/kg, 3.0 mg/kg, 3.1 mg/kg, 3.2 mg/kg, 3.3 mg/kg, 3.4 mg/kg, 3.5 mg/kg, 3.6 mg/kg, 3.7 mg/kg, 3.8 mg/kg, 3.9 mg/kg, 4 mg/kg
  • a CFB dsRNA agent of the invention may be considered in the determination of dosage and timing of delivery of a CFB dsRNA agent of the invention.
  • the absolute amount of a CFB dsRNA agent or CFB antisense polynucleotide agent delivered will depend upon a variety of factors including a concurrent treatment, the number of doses and the individual subject parameters including age, physical condition, size and weight. These are factors well known to those of ordinary skill in the art and can be addressed with no more than routine experimentation.
  • a maximum dose can be used, that is, the highest safe dose according to sound medical judgment.
  • Methods of the invention may in some embodiments include administering to a subject 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more doses of a CFB dsRNA agent or CFB antisense polynucleotide agent.
  • a pharmaceutical compound e.g., comprising a CFB dsRNA agent or comprising a CFB antisense polynucleotide agent
  • Doses may be administered once per day or more than once per day, for example, 2, 3, 4, 5, or more times in one 24 hour period.
  • a pharmaceutical composition of the invention may be administered once daily, or the CFB dsRNA agent or CFB antisense polynucleotide agent may be administered as two, three, or more sub-doses at appropriate intervals throughout the day or even using continuous infusion or delivery through a controlled release formulation.
  • a pharmaceutical composition of the invention is administered to a subject one or more times per day, one or more times per week, one or more times per month, or one or more times per year.
  • Methods of the invention include administration of a pharmaceutical compound alone, in combination with one or more other CFB dsRNA agents or CFB antisense polynucleotide agents, and/or in combination with other drug therapies or treatment activities or regimens that are administered to subjects with a CFB-associated disease or condition.
  • Pharmaceutical compounds may be administered in pharmaceutical compositions.
  • Pharmaceutical compositions used in methods of the invention may be sterile and contain an amount of a CFB dsRNA agent or CFB antisense polynucleotide agent that will reduce activity of a CFB polypeptide to a level sufficient to produce the desired response in a unit of weight or volume suitable for administration to a subject.
  • a dose administered to a subject of a pharmaceutical composition that includes a CFB dsRNA agent or CFB antisense polynucleotide agent to reduce CFB protein activity can be chosen in accordance with different parameters, in particular in accordance with the mode of administration used and the state of the subject. Other factors include the desired period of treatment. In the event that a response in a subject is insufficient at the initial doses applied, higher doses (or effectively higher doses by a different, more localized delivery route) may be employed to the extent that patient tolerance permits.
  • CFB-associated disease As used herein, “CFB-associated disease” , “CFB-associated diseases and conditions” and “diseases and conditions caused and/or modulated by CFB” is intended to include any disease associated with the CFB gene or protein. Such diseases may be caused, for example, by overproduction of CFB protein, by mutation of the CFB gene, by abnormal cleavage of the CFB protein, by abnormal interaction between CFB and other proteins or other endogenous or exogenous substances.
  • Exemplary CFB -associated diseases include, but are not limited to: autoimmune diseases, complement system dysfunction including aberrant upregulation of complement components such as CFB, C3 glomerulopathy (C3G) , systemic lupus erythematosus (SLE) , Lupus Nephritis, Ig-mediated kidney pathologies such as IgA nephropathy and primary membranous nephropathy, nephropathy, diabetic nephropathy, polycystic kidney disease, membranous nephropathy, age-related macular degeneration (AMD) including dry AMD and geographic atrophy, typical or infectious hemolytic uremic syndrome (tHUS) , atypical hemolytic uremic syndrome (aHUS) , asthma, psoriasis, thrombotic microangiopathy, ischemia and reperfusion injury, paroxysmal nocturnal hemoglobinuria (PNH) , rheumatic disease, rheumatoid arthritis, multiple
  • coli-related hemolytic uremic syndrome myasthenia gravis (MG) , neuromyelistis optica (NMO) , dense deposit disease, Coronary artery disease, dermatomyositis, Graves' disease, atherosclerosis, Alzheimer's disease, systemic inflammatory response sepsis, septic shock, spinal cord injury, glomerulonephritis, Hashimoto's thyroiditis, type I diabetes, psoriasis, pemphigus, autoimmune hemolytic anemia (AIHA) , cold agglutinin disease, humoral and vascular transplant rejection, graft dysfunction, myocardial infarction, sensitization towards a transplant, hyperlipidemia, and sepsis, or lipid imbalance related to CFB.
  • MG myasthenia gravis
  • NMO neuromyelistis optica
  • a subject may be administered a CFB dsRNA agent or CFB antisense polynucleotide agent of the invention at a time that is one or more of before or after diagnosis of a CFB-associated disease or condition.
  • a subject is at risk of having or developing a CFB-associated disease or condition.
  • a subject at risk of developing a CFB-associated disease or condition is one who has an increased probability of developing the CFB-associated disease or condition, compared to a control risk of developing the CFB-associated disease or condition.
  • a level of risk may be statistically significant compared to a control level of risk.
  • a subject at risk may include, for instance, a subject who is, or will be, a subject who has a preexisting disease and/or a genetic abnormality that makes the subject more susceptible to a CFB-associated disease or condition than a control subject without the preexisting disease or genetic abnormality; a subject having a family and/or personal medical history of the CFB-associated disease or condition; and a subject who has previously been treated for a CFB-associated disease or condition.
  • a preexisting disease and/or a genetic abnormality that makes the subject more susceptible to a CFB-associated disease or condition may be a disease or genetic abnormality that when present has been previously identified as having a correlative relation to a higher likelihood of developing a CFB-associated disease or condition.
  • a CFB dsRNA agent or CFB antisense polynucleotide agent may be administered to a subject based on a medical status of the individual subject.
  • a health-care provided for a subject may assess a CFB level measured in a sample obtained from a subject and determine it is desirable to reduce the subject’s CFB level, by administration of a CFB dsRNA agent or CFB antisense polynucleotide agent of the invention.
  • the CFB level may be considered to be a physiological characteristic of a CFB-associated condition, even if the subject is not diagnosed as having a CFB-assoicated disease such as one disclosed herein.
  • a healthcare provider may monitor changes in the subject’s CFB level, as a measure of efficacy of the administered CFB dsRNA agent or CFB antisense polynucleotide agent of the invention.
  • a biological sample such as a blood or serum sample may be obtained from a subject and a CFB level for the subject determined in the sample.
  • a CFB dsRNA agent or CFB antisense polynucleotide agent is administered to the subject and a blood sample is obtained from the subject following the administration and the CFB level determined using the sample and the results compared to the results determined in the subject’s pre-administration (prior) sample.
  • a reduction in the subject’s CFB level in the later sample compared to the pre-administration level indicates the administered CFB dsRNA agent or CFB antisense polynucleotide agent efficacy in reducing the lipid level in the subject.
  • Certain embodiments of methods of the invention include adjusting a treatment that includes administering a dsRNA agent or a CFB antisense polynucleotide agent of the invention to a subject based at least in part on assessment of a change in one or more of the subject’s physiological characteristics of a CFB-associated disease or condition resulting from the treatment.
  • an effect of an administered dsRNA agent or CFB antisense polynucleotide agent of the invention may be determined for a subject and used to assist in adjusting an amount of a dsRNA agent or CFB antisense polynucleotide agent of the invention subsequently administered to the subject.
  • a subject is administered a dsRNA agent or CFB antisense polynucleotide agent of the invention, the subject’s CFB level is determined after the administration, and based at least in part on the determined level, a greater amount of the dsRNA agent or CFB antisense polynucleotide agent is determined to be desirable in order to increase the physiological effect of the administered agent, for example to reduce or further reduce the subject’s CFB level.
  • a subject is administered a dsRNA agent or CFB antisense polynucleotide agent of the invention, the subject’s CFB level is determined after the administration and based at least in part on the determined level, a lower amount of the dsRNA agent or CFB antisense polynucleotide agent is desirable to administer to the subject.
  • some embodiments of the invention include assessing a change in one or more physiological characteristics of resulting from a subject’s previous treatment to adjust an amount of a dsRNA agent or CFB antisense polynucleotide agent of the invention subsequently administered to the subject.
  • Some embodiments of methods of the invention include 1, 2, 3, 4, 5, 6, or more determinations of a physiological characteristic of a CFB-associated disease or condition to assess and/or monitor the efficacy of an administered CFB dsRNA agent or CFB antisense polynucleotide agent of the invention, and optionally using the determinations to adjust one or more of: a dose, administration regimen, and or administration frequency of a dsRNA agent or CFB antisense polynucleotide agent of the invention to treat a CFB-associated disease or condition in a subject.
  • a desired result of administering an effective amount of a dsRNA agent or CFB antisense polynucleotide agent of the invention to a subject is a reduction of the subject’s the CFB mRNA level, the CFB protein level in the subject, or CH50 activity (a measure of total hemolytic complement) , AH50 (a measure the hemolytic activity of the alternate pathway of complement) , lactate dehydrogenase (LDH) (a measure of intravascular hemolysis) , hemoglobin levels; the level of any one or more of C3, C9, C5, C5a, C5b, and soluble C5b-9 complex.
  • the terms “treat” , “treated” , or “treating” when used with respect to a CFB-associated disease or condition may refer to a prophylactic treatment that decreases the likelihood of a subject developing the CFB-associated disease or condition, and also may refer to a treatment after the subject has developed a CFB-associated disease or condition in order to eliminate or reduce the level of the CFB-associated disease or condition, prevent the CFB-associated disease or condition from becoming more advanced (e.g., more severe) , and/or slow the progression of the CFB-associated disease or condition in a subject compared to the subject in the absence of the therapy to reduce activity in the subject of CFB polypeptide.
  • agents, compositions, and methods of the invention can be used to inhibit CFB gene expression.
  • the terms “inhibit, ” “silence, ” “reduce, ” “down-regulate, ” and “knockdown” mean the expression of the CFB gene, as measured by one or more of: a level of RNA transcribed from the gene, a level of activity of CFB expressed, and a level of CFB polypeptide, protein or protein subunit translated from the mRNA in a cell, group of cells, tissue, organ, or subject in which the CFB gene is transcribed, is reduced when the cell, group of cells, tissue, organ, or subject is contacted with (e.g., treated with) a CFB dsRNA agent or CFB antisense polynucleotide agent of the invention, compared to a control level of RNA transcribed from the CFB gene, a level of activity of expressed CFB, or a level of CFB dsRNA agent or CFB antisense polynucle
  • a variety of administration routes for a CFB dsRNA agent or CFB antisense polynucleotide agent are available for use in methods of the invention.
  • the particular delivery mode selected will depend at least in part, upon the particular condition being treated and the dosage required for therapeutic efficacy. Methods of this invention, generally speaking, may be practiced using any mode of administration that is medically acceptable, meaning any mode that produces effective levels of treatment of a CFB-associated disease or condition without causing clinically unacceptable adverse effects.
  • a CFB dsRNA agent or CFB antisense polynucleotide agent may be administered via an oral, enteral, mucosal, subcutaneous, and/or parenteral route.
  • parenteral includes subcutaneous, intravenous, intrathecal, intramuscular, intraperitoneal, and intrasternal injection, or infusion techniques.
  • Other routes include but are not limited to nasal (e.g., via a gastro-nasal tube) , dermal, vaginal, rectal, sublingual, and inhalation.
  • Delivery routes of the invention may include intrathecal, intraventricular, or intracranial.
  • a CFB dsRNA agent or CFB antisense polynucleotide agent may be placed within a slow-release matrix and administered by placement of the matrix in the subject.
  • a CFB dsRNA agent or CFB antisense polynucleotide agent may be delivered to a subject cell using nanoparticles coated with a delivery agent that targets a specific cell or organelle.
  • a delivery agent that targets a specific cell or organelle.
  • Various delivery means, methods, agents are known in the art. Non-limiting examples of delivery methods and delivery agents are additionally provided elsewhere herein.
  • the term “delivering” in reference to a CFB dsRNA agent or CFB antisense polynucleotide agent may mean administration to a cell or subject of one or more “naked” CFB dsRNA agent or CFB antisense polynucleotide agent sequences and in certain aspects of the invention “delivering” means administration to a cell or subject via transfection means, delivering a cell comprising a CFB dsRNA agent or CFB antisense polynucleotide agent to a subject, delivering a vector encoding a CFB dsRNA agent or CFB antisense polynucleotide agent into a cell and/or subject, etc. Delivery of a CFB dsRNA agent or CFB antisense polynucleotide agent using a transfection means may include administration of a vector to a cell and/or subject.
  • one or more CFB dsRNA agents or CFB antisense polynucleotide agents may be administered in formulations, which may be administered in pharmaceutically acceptable solutions, which may routinely contain pharmaceutically acceptable concentrations of salt, buffering agents, preservatives, compatible carriers, adjuvants, and optionally other therapeutic ingredients.
  • a CFB dsRNA agent or CFB antisense polynucleotide agent may be formulated with another therapeutic agent for simultaneous administration.
  • a CFB dsRNA agent or CFB antisense polynucleotide agent may be administered in a pharmaceutical composition.
  • a pharmaceutical composition comprises a CFB dsRNA agent or CFB antisense polynucleotide agent and optionally, a pharmaceutically acceptable carrier.
  • Pharmaceutically acceptable carriers are well-known to those of ordinary skill in the art.
  • a pharmaceutically acceptable carrier means a non-toxic material that does not interfere with the effectiveness of the biological activity of the active ingredients, e.g., the ability of the CFB dsRNA agent or CFB antisense polynucleotide agent to inhibit CFB gene expression in a cell or subject. Numerous methods to administer and deliver dsRNA agents or CFB antisense polynucleotide agents for therapeutic use are known in the art and may be utilized in methods of the invention.
  • Pharmaceutically acceptable carriers include diluents, fillers, salts, buffers, stabilizers, solubilizers and other materials that are well-known in the art. Exemplary pharmaceutically acceptable carriers are described in U.S. Pat. No. 5,211,657 and others are known by those skilled in the art. Such preparations may routinely contain salt, buffering agents, preservatives, compatible carriers, and optionally other therapeutic agents. When used in medicine, the salts should be pharmaceutically acceptable, but non-pharmaceutically acceptable salts may conveniently be used to prepare pharmaceutically acceptable salts thereof and are not excluded from the scope of the invention.
  • Such pharmacologically and pharmaceutically acceptable salts include, but are not limited to, those prepared from the following acids: hydrochloric, hydrobromic, sulfuric, nitric, phosphoric, maleic, acetic, salicylic, citric, formic, malonic, succinic, and the like.
  • pharmaceutically acceptable salts can be prepared as alkaline metal or alkaline earth salts, such as sodium, potassium or calcium salts.
  • Some embodiments of methods of the invention include administering one or more CFB dsRNA agents or CFB antisense polynucleotide agents directly to a tissue.
  • the tissue to which the compound is administered is a tissue in which the CFB-associated disease or condition is present or is likely to arise, non-limiting examples of which are the heart.
  • Direct tissue administration may be achieved by direct injection or other means. Many orally delivered compounds naturally travel to and through the liver and kidneys and some embodiments of treatment methods of the invention include oral administration of one or more CFB dsRNA agents to a subject.
  • CFB dsRNA agents or CFB antisense polynucleotide agents may be administered once, or alternatively they may be administered in a plurality of administrations. If administered multiple times, the CFB dsRNA agent or CFB antisense polynucleotide agent may be administered via different routes. For example, though not intended to be limiting, a first (or first several) administrations may be made via subcutaneous means and one or more additional administrations may be oral and/or systemic administrations.
  • the CFB dsRNA agent or CFB antisense polynucleotide agent may be formulated for parenteral administration by injection, e.g., by bolus injection or continuous infusion.
  • Formulations for injection may be presented in unit dosage form, e.g., in ampoules or in multi-dose containers, with or without an added preservative.
  • CFB dsRNA agent formulations (also referred to as pharmaceutical compositions) may take such forms as suspensions, solutions or emulsions in oily or aqueous vehicles, and may contain formulatory agents such as suspending, stabilizing and/or dispersing agents.
  • Preparations for parenteral administration include sterile aqueous or non-aqueous solutions, suspensions, and emulsions.
  • non-aqueous solvents are propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable organic esters such as ethyl oleate.
  • Aqueous carriers include water, alcoholic/aqueous solutions, emulsions or suspensions, including saline and buffered media.
  • Parenteral vehicles include sodium chloride solution, Ringer's dextrose, dextrose and sodium chloride, lactated Ringer's , or fixed oils.
  • Intravenous vehicles include fluid and nutrient replenishers, electrolyte replenishers (such as those based on Ringer's dextrose) , and the like. Preservatives and other additives may also be present such as, for example, antimicrobials, anti-oxidants, chelating agents, and inert gases and the like. Lower doses will result from other forms of administration, such as intravenous administration. In the event that a response in a subject is insufficient at the initial doses applied, higher doses (or effectively higher doses by a different, more localized delivery route) may be employed to the extent that patient tolerance permits. Multiple doses per day may be used as needed to achieve appropriate systemic or local levels of one or more CFB dsRNA agents or CFB antisense polynucleotide agents and to achieve appropriate reduction in CFB protein activity.
  • methods of the invention include use of a delivery vehicle such as biocompatible microparticle, nanoparticle, or implant suitable for implantation into a recipient, e.g., a subject.
  • a delivery vehicle such as biocompatible microparticle, nanoparticle, or implant suitable for implantation into a recipient, e.g., a subject.
  • exemplary bioerodible implants that may be useful in accordance with this method are described in PCT Publication No. WO 95/24929 (incorporated by reference herein) , which describes a biocompatible, biodegradable polymeric matrix for containing a biological macromolecule.
  • a matrix may be biodegradable.
  • Matrix polymers may be natural or synthetic polymers.
  • a polymer can be selected based on the period of time over which release is desired, generally in the order of a few hours to a year or longer. Typically, release over a period ranging from between a few hours and three to twelve months can be used.
  • the polymer optionally is in the form of a hydrogel that can absorb up to about 90%of its weight in water and further, optionally is cross-linked with multivalent ions or other polymers.
  • CFB dsRNA agents or CFB antisense polynucleotide agents may be delivered in some embodiments of the invention using the bioerodible implant by way of diffusion, or by degradation of the polymeric matrix.
  • Exemplary synthetic polymers for such use are well known in the art.
  • Biodegradable polymers and non-biodegradable polymers can be used for delivery of CFB dsRNA agents or CFB antisense polynucleotide agents using art-known methods.
  • Bioadhesive polymers such as bioerodible hydrogels (see H.S. Sawhney, C.P. Pathak and J.A.
  • Hubell in Macromolecules, 1993, 26, 581-587, the teachings of which are incorporated by reference herein) may also be used to deliver CFB dsRNA agents or CFB antisense polynucleotide agents for treatment of a CFB-associated disease or condition.
  • Additional suitable delivery systems can include time-release, delayed release or sustained release delivery systems. Such systems can avoid repeated administrations of a CFB dsRNA agent or CFB antisense polynucleotide agent, increasing convenience to the subject and the medical care professional.
  • Many types of release delivery systems are available and known to those of ordinary skill in the art. (See for example: U.S. Pat. Nos.
  • pump-based hardware delivery systems can be used, some of which are adapted for implantation.
  • Long-term sustained release implant may be suitable for prophylactic treatment of subjects and for subjects at risk of developing a recurrent CFB-associated disease or condition.
  • Long-term release means that the implant is constructed and arranged to deliver a therapeutic level of a CFB dsRNA agent or CFB antisense polynucleotide agent for at least up to 10 days, 20 days, 30 days, 60 days, 90 days, six months, a year, or longer.
  • Long-term sustained release implants are well-known to those of ordinary skill in the art and include some of the release systems described above.
  • Therapeutic formulations of CFB dsRNA agents or CFB antisense polynucleotide agents may be prepared for storage by mixing the molecule or compound having the desired degree of purity with optional pharmaceutically acceptable carriers, excipients or stabilizers [Remington's Pharmaceutical Sciences 21 st edition, (2006) ] , in the form of lyophilized formulations or aqueous solutions.
  • Acceptable carriers, excipients, or stabilizers are nontoxic to recipients at the dosages and concentrations employed, and include buffers such as phosphate, citrate, and other organic acids; antioxidants including ascorbic acid and methionine; preservatives (such as octadecyldimethylbenzyl ammonium chloride; hexamethonium chloride; benzalkonium chloride, benzethonium chloride; phenol, butyl or benzyl alcohol; alkyl parabens such as methyl or propyl paraben; catechol; resorcinol; cyclohexanol; 3-pentanol; and m-cresol) ; low molecular weight (less than about 10 residues) polypeptides; proteins, such as serum albumin, gelatin, or immunoglobulins; hydrophilic polymers such as polyvinylpyrrolidone; amino acids such as glycine, glutamine, asparagine, histidine
  • Methods of the invention may be used in conjunction with cells, tissues, organs and/or subjects.
  • a subject is a human or vertebrate mammal including but not limited to a dog, cat, horse, cow, goat, mouse, rat, and primate, e.g., monkey.
  • the invention can be used to treat CFB-associated diseases or conditions in human and non-human subjects.
  • a subject may be a farm animal, a zoo animal, a domesticated animal or non-domesticated animal and methods of the invention can be used in veterinary prevention and treatment regimens.
  • the subject is a human and methods of the invention can be used in human prevention and treatment regimens.
  • Non-limiting examples of subjects to which the present invention can be applied are subjects who are diagnosed with, suspected of having, or at risk of having a disease or condition associated with a higher than desirable CFB expression and/or activity, also referred to as “elevated levels of CFB expression” .
  • elevated levels of CFB expression Non-limiting examples of diseases and conditions associated with a higher than desirable levels of CFB expression and/or activity are described elsewhere herein.
  • Methods of the invention may be applied to a subject who, at the time of treatment, has been diagnosed as having the disease or condition associated with a higher than desirable CFB expression and/or activity, or a subject who is considered to be at risk for having or developing a disease or condition associated with a higher than desirable CFB expression and/or activity.
  • a disease or condition associated with a higher than desirable CFB level of expression and/or activity is an acute disease or condition, and in certain aspects of the invention a disease or condition associated with a higher than desirable CFB level of expression and/or activity is a chronic disease or condition.
  • a CFB dsRNA agent of the invention is administered to a subject diagnosed with, suspected of having, or at risk of having, statin resistant hypercholesterolemia, which is a disease in which it is desirable to reduce CFB expression.
  • Methods of the invention may be applied to the subject who, at the time of treatment, has been diagnosed as having the disease or condition, or a subject who is considered to be at risk for having or developing the disease or condition.
  • a CFB dsRNA agent of the invention is administered to a subject diagnosed with, suspected of having, or at risk of having, hyperlipidemia, which is a disease in which it is desirable to reduce CFB expression.
  • Methods of the invention may be applied to the subject who, at the time of treatment, has been diagnosed as having the disease or condition, or a subject who is considered to be at risk for having or developing the disease or condition.
  • a cell to which methods of the invention may be applied include cells that are in vitro, in vivo, ex vivo cells. Cells may be in a subject, in culture, and/or in suspension, or in any other suitable state or condition.
  • a cell to which a method of the invention may be applied can be a liver cell, a hepatocyte, a cardiac cell, a pancreatic cell, a cardiovascular cell, kidney cell or other type of vertebrate cell, including human and non-human mammalian cells.
  • a cell to which methods of the invention may be applied is a healthy, normal cell that is not known to be a disease cell.
  • a control cell is a normal cell, but it will be understood that a cell having a disease or condition may also serve as a control cell in particular circumstances for example to compare results in a treated cell having a disease or condition versus an untreated cell having the disease or condition, etc.
  • a level of CFB polypeptide activity can be determined and compared to control level of CFB polypeptide activity, according to methods of the invention.
  • a control may be a predetermined value, which can take a variety of forms. It can be a single cut-off value, such as a median or mean. It can be established based upon comparative groups, such as in groups having normal levels of CFB polypeptide and/or CFB polypeptide activity and groups having increased levels of CFB polypeptide and/or CFB polypeptide activity.
  • comparative groups may be groups having one or more symptoms of or a diagnosis of a CFB-associated disease or condition; groups without having one or more symptoms of or a diagnosis of the disease or condition; groups of subjects to whom an siRNA treatment of the invention has been administered; groups of subjects to whom an siRNA treatment of the invention has not been administered.
  • a control may be based on apparently healthy normal individuals in an appropriate age bracket or apparently healthy cells. It will be understood that controls according to the invention may be, in addition to predetermined values, samples of materials tested in parallel with the experimental materials. Examples include samples from control populations or control samples generated through manufacture to be tested in parallel with the experimental samples.
  • a control may include a cell or subject not contacted or treated with a CFB dsRNA agent of the invention and in such instances, a control level of CFB polypeptide and/or CFB polypeptide activity can be compared to a level of CFB polypeptide and/or CFB polypeptide activity in a cell or subject contacted with a CFB dsRNA agent or CFB antisense polynucleotide agent of the invention.
  • a level of CFB polypeptide determined for a subject can be a control level against which a level of CFB polypeptide determined for the same subject at a different time is compared.
  • a level of CFB is determined in a biological sample obtained from a subject who has not been administered a CFB treatment of the invention.
  • the biological sample is a serum sample.
  • the level of CFB polypeptide determined in the sample obtained from the subject can serve as a baseline or control value for the subject.
  • one or more additional serum samples can be obtained from the subject and the level of CFB polypeptide in the subsequent sample or samples can be compared to the control/baseline level for the subject. Such comparisons can be used to assess onset, progression, or recession of a CFB associated disease or condition in the subject.
  • a level of CFB polypeptide in the baseline sample obtained from the subject that is higher than a level obtained from the same subject after the subject has been administered a CFB dsRNA agent or CFB antisense polynucleotide agent of the invention indicates regression of the CFB-associated disease or condition and indicates efficacy of the administered CFB dsRNA agent of the invention for treatment of the CFB-associated disease or condition.
  • values of one or more of a level of CFB polypeptide and/or CFB polypeptide activity determined for a subject may serve as control values for later comparison of levels of CFB polypeptide and/or CFB activity, in that same subject, thus permitting assessment of changes from a “baseline” CFB polypeptide activity in a subject.
  • an initial CFB polypeptide level and/or initial CFB polypeptide activity level may be present and/or determined in a subject and methods and compounds of the invention may be used to decrease the level of CFB polypeptide and/or CFB polypeptide activity in the subject, with the initial level serving as a control level for that subject.
  • CFB dsRNA agents and/or CFB antisense polynucleotide agents of the invention may be administered to a subject.
  • Efficacy of the administration and treatment of the invention can be assessed when a level of CFB polypeptide in a serum sample obtained from a subject is decreased by at least 0.5%, 1%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, or more compared to a pre-administration level of CFB polypeptide in a serum sample obtained from the subject at a prior time point, or compared to a non-contacted control level, for example a level of CFB polypeptide in a control serum sample.
  • Certain embodiments of methods of the invention comprise administering a CFB dsRNA and/or CFB antisense agent of the invention to a subject in an amount effective to inhibit CFB gene expression and thereby reduce a level of CFB polypeptide and reduce a level of CFB polypeptide activity in the subject.
  • Some embodiments of the invention include determining presence, absence, and/or an amount (also referred to herein as a level) of CFB polypeptide in one or more biological samples obtained from one or more subjects. The determination can be used to assess efficacy of a treatment method of the invention. For example, methods and compositions of the invention can be used to determine a level of CFB polypeptide in a biological sample obtained from a subject previously treated with administration of a CFB dsRNA agent and/or a CFB antisense agent of the invention.
  • a physiological characteristic of a CFB-associated disease or condition determined for a subject can be a control determination against which a determination of the physiological characteristic in the same subject at a different time is compared.
  • a physiological characteristic such as the CFB mRNA level, the CFB protein level in the subject, or CH50 activity , AH50 , lactate dehydrogenase (LDH) , hemoglobin levels; the level of any one or more of C3, C9, C5, C5a, C5b, and soluble C5b-9 complex in the plasma or the tissue sample is determined in a biological sample, such as a serum sample, obtained from a subject who has not been administered a CFB treatment of the invention.
  • a biological sample such as a serum sample
  • the CFB mRNA level (and/or other physiological characteristic of a CFB disease or condition) determined in the sample obtained from the subject can serve as a baseline or control value for the subject.
  • a CFB dsRNA agent to the subject in a treatment method of the invention
  • one or more additional serum samples can be obtained from the subject and CFB mRNA level and/or CFB protein level in the subsequent sample or samples are compared to the control/baseline level and/or ratio, respectively, for the subject. Such comparisons can be used to assess onset, progression, or recession of a CFB associated disease or condition in the subject.
  • CFB mRNA level in the baseline sample obtained from the subject that is higher than CFB mRNA level determined in a sample obtained from the same subject after the subject has been administered a CFB dsRNA agent or CFB antisense polynucleotide agent of the invention indicates regression of the CFB-associated disease or condition and indicates efficacy of the administered CFB dsRNA agent of the invention for treatment of the CFB-associated disease or condition.
  • values of one or more of a physiological characteristic of a CFB-associcated disease or condition determined for a subject may serve as control values for later comparison of the physiological characteristics in that same subject, thus permitting assessment of changes from a “baseline” physiological characteristic in a subject.
  • an initial physiological characteristic may be present and/or determined in a subject and methods and compounds of the invention may be used to decrease the level of CFB polypeptide and/or CFB polypeptide activity in the subject, with the initial physiological characteristic determination serving as a control for that subject.
  • CFB dsRNA agents and/or CFB antisense polynucleotide agents of the invention may be administered to a subject in an effective amount to treat a CFB disease or condition.
  • Efficacy of the administration and treatment of the invention can be assessed by determining a change in one or more physiological characteristics of the CFB disease or condition.
  • a CFB mRNA level in a serum sample obtained from a subject is decreased by at least 0.5%, 1%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, or more compared to a pre-administration lipid in a serum sample obtained from the subject at a prior time point, or compared to a non-contacted control level, for example CFB mRNA level in a control serum sample.
  • a CFB mRNA level in a serum sample obtained from a subject is decreased by at least 0.5%, 1%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, or more compared to a pre-administration lipid in a serum sample obtained from the subject at a prior time point, or compared to a non-contacted control level, for example CFB mRNA level in a control serum sample.
  • Certain embodiments of methods of the invention comprise administering a CFB dsRNA and/or CFB antisense agent of the invention to a subject in an amount effective to inhibit CFB gene expression and thereby reduce the CFB mRNA level, the CFB protein level in the subject, or otherwise positively impact a physiological characteristic of a CFB-assocaited disease or condition in the subject.
  • Some embodiments of the invention include determining presence, absence, and/or a change in a physiological characteristic of a CFB-associated disease or condition using methods such as but not limited to: (1) assessing one or more biological samples obtained from one or more subjects for the physiological characteristic; (2) imaging a subject (for example but not limited to obtaining a liver image) ; and (3) or physical examination of the subject. The determination can be used to assess efficacy of a treatment method of the invention.
  • kits that comprise one or more CFB dsRNA agents and/or CFB antisense polynucleotide agents and instructions for its use in methods of the invention.
  • Kits of the invention may include one or more of a CFB dsRNA agent, CFB sense polynucleotide, and CFB antisense polynucleotide agent that may be used to treat a CFB-associated disease or condition.
  • Kits containing one or more CFB dsRNA agents, CFB sense polynucleotides, and CFB antisense polynucleotide agents can be prepared for use in treatment methods of the invention.
  • Components of kits of the invention may be packaged either in aqueous medium or in lyophilized form.
  • a kit of the invention may comprise a carrier being compartmentalized to receive in close confinement therein one or more container means or series of container means such as test tubes, vials, flasks, bottles, syringes, or the like.
  • a first container means or series of container means may contain one or more compounds such as a CFB dsRNA agent and/or CFB sense or antisense polynucleotide agent.
  • a second container means or series of container means may contain a targeting agent, a labelling agent, a delivery agent, etc. that may be included as a portion of a CFB dsRNA agent and/or CFB antisense polynucleotide to be administered in an embodiment of a treatment method of the invention.
  • a kit of the invention may also include instructions. Instructions typically will be in written form and will provide guidance for carrying-out a treatment embodied by the kit and for making a determination based upon that treatment.
  • phosphoramidites may be prepared according to procedures described herein and/or prior arts such as, but are not limited to, US426, 220 and WO02/36743.
  • Dichloromethane (19.50kg) was added to the 50 L glass kettle under the protection of nitrogen and started stirring.
  • the temperature was controlled at 20 ⁇ 30 °C, and DMTr imann (1.47 kg) , triethylamine (1.50 kg) , 4-dimethylaminopyridine (0.164 kg) and succinic anhydride (1.34 kg) was added to the glass kettle.
  • the system was kept at 20 ⁇ 30 °C for 18h, samples were taken and the reaction was ended.
  • Saturated sodium bicarbonate solution (22.50 kg) was added into the reaction system, stirred for 10-20 min, and allowed to separate into layers.
  • the organic phase was separated, and the aqueous phase was extracted twice with dichloromethane, and the organic phase was combined and dried over anhydrous sodium sulfate, filtered, and concentrated in vacuum to get the residue forming a gray to off-white solid of 1.83 kg.
  • N, N-dimethylformamide (23.50 kg) was added into a 100L glass kettle and stirred. The temperature was controlled at 20 ⁇ 30 °C. Under the protection of nitrogen, the products of the previous step, O-benzotriazole tetramethylurea hexafluorophosphate (0.33 kg) and N, N-diisopropylethylamine (0.13 kg) were added into the aforesaid 100L glass kettle through the solid feeding funnel and stirred for 10 ⁇ 30 minutes and were discharged into a 50 L zinc barrel for use.
  • Macroporous amine methyl resin (3.25 kg) (purchased from Tianjin Nankai Hecheng Science and Technology Co., Ltd., batch number HA2X1209, load capacity 0.48 mmol/g) were added into the aforesaid 100 L solid phase synthesis reactor through the solid feeding funnel, the temperature was controlled at 20 ⁇ 30 °C, N, N-dimethylformamide (21.00 kg+21.00 kg) and the reaction solution in the zinc barrel of the previous step were add into the solid phase synthesis reactor. The system was subject to thermal insulation reaction, and the solid load was tracked to ⁇ 250umol/g, and the load detection method was UV.
  • the isomannide residue can be added to the 5'-end or 3'-end of the oligonucleotide chain by a method well known to those skilled in the art, such as the reverse abasic (invab) method, and further added to the target to the group.
  • a method well known to those skilled in the art such as the reverse abasic (invab) method
  • Sense and antisense strand sequences of siRNA were synthesized on oligonucleotide synthesizers using a well-established solid phase synthesis method based on phosphoramidite chemistry. Oligonucleotide chain propagation is achieved through 4-step cycles: a deprotection, a condensation, a capping and an oxidation or a sulfurization step for addition of each nucleotide. Syntheses were performed on a solid support made of controlled pore glass (CPG, ) . Monomer phosphoramidites may be purchased from commercial sources or may be the phosporamidite compounds in example 1.
  • the phosporamidite compounds herein may be attached to the 3'-end as a monomeric phosphoramidite, and further be attached to the CPG solid support. In the case of attachment at the 5'-end, the phosphoramidite compounds may be used for the final coupling reaction, and can be further conjugated to target ligands if necessary.
  • Phosphoramidites with GalNAc ligand cluster were public in WO2023/045995A1 (incorporated herein in its entirety) .
  • GalNAc ligand (GLS-5*or GLS-15*as non-limiting example are attached at 5’-end of sense strand
  • GLS-5*or GLS-15*Phosphoramidites with GalNAc ligand cluster were public in WO2023/045995A1 (incorporated herein in its entirety)
  • a GalNAc phosphoramidite was used for the last coupling reaction.
  • Trichloroacetic acid (TCA) 3%in dichloromethane was used for deprotection of 4, 4′-dimethoxytrityl protecting group (DMT) .
  • DMT 4′-dimethoxytrityl protecting group
  • 5-Ethylthio-1H-tetrazole was used as an activator.
  • IP-RP-HPLC ion pairing reversed phase HPLC
  • Purified single strand oligonucleotide product from IP-RP-HPLC was converted to sodium salt by dissolving in 1.0 M NaOAc and precipitation by addition of ice cold EtOH. Annealing of equimolar complementary sense stand and antisense strand oligonucleotide in water was performed to form the double strand siRNA product, which was lyophilized to afford a fluffy white solid.
  • a method used to attach a targeting group comprising GalNAc (also referred to herein as a GalNAc delivery compound) to the 5’-end of a sense strand included use of a GalNAc phosphoramidite (GLS-5*or GLS-15*Phosphoramidites) in the last coupling step in the solid phase synthesis, using a synthetic process such as the process used if oligonucleotide chain propagation of adding a nucleotide to the 5’-end of the sense strand is performed.
  • GalNAc phosphoramidite GLS-5*or GLS-15*Phosphoramidites
  • a method of attaching a targeting group comprising GalNAc to the 3’-end of a sense strand comprised use of a solid support (CPG) that included a GLO-n.
  • a method of attaching a targeting group comprising GalNAc to the 3’-end of a sense strand comprises attaching a GalNAc targeting group to CPG solid support through an ester bond and using the resulting CPG with the attached GalNAc targeting group when synthesizing the sense strand, which results in the GalNAc targeting group attached at the 3’-end of the sense strand.
  • the imann residues can be added to the 5' end or 3' end of the oligonucleotide chain by a method well known to those skilled in the art, such as the inverted abasic residues (invab) method, and/or further added to the target to the GalNAc targeting group.
  • invab inverted abasic residues
  • Huh7 cells were trypsinized and adjusted to appropriate density, mixed with the complexes of psiCHECK (TM) -2 Vector plasmid and Lipofectamine 2000 (Invitrogen-11668-019) and seeded into 96-well plates.
  • Cells were transfected with test siRNAs or a control siRNA using Lipofectamine RNAiMax (Invitrogen -13778-150) at the same time of seeding following the protocol according to manufacturer’s recommendation.
  • the siRNAs were tested at two concentrations (1 nM and 10 nM) in triplicate.
  • No compound control well was defined as cells transfected with psiCHECK (TM) -2 Vector and without siRNA treatment; blank control was cell only wells.
  • Ratio of sample well (sample Renilla luminescence-background blank) / (sample Fireflyluminescence-background blank)
  • Ratio of no compound control well (control Renilla luminescence-background blank) / (control sample Fireflyluminescence-background blank)
  • duplex sequences used correspond to those shown in Table 5-7.
  • Table 5 provides experimental results of in vitro studies using various CFB RNAi agents to inhibit CFB expression.
  • the duplex sequences used correspond to those shown in Table 2.
  • Table 6 provides experimental results of in vitro studies using various CFB RNAi agents to inhibit CFB expression.
  • the duplex sequences used correspond to those shown in Table 2.
  • Table 7 provides experimental results of in vitro studies using various CFB RNAi agents to inhibit CFB expression.
  • the duplex sequences used correspond to those shown in Table 2.
  • mice Female C57BL/6J mice (4 in each group) were infected by intravenous administration of a solution of adeno-associated virus 8 (AAV8) vector encoding human CFB and luciferase gene.
  • AAV8 adeno-associated virus 8
  • mice were subcutaneously administered a single 2 or 3 mg/kg of CFB siRNA agents or PBS.
  • Blood samples were collected at day 8, before dosing of siRNA, at day 15, day 22 and day29. Plasma samples were isolated and luciferase activity of plasma samples was measured per manufacturer’s recommended protocol.
  • mice Female C57BL/6J mice (4 in each group) were infected by intravenous administration of a solution of adeno-associated virus 8 (AAV8) vector encoding human CFB and luciferase gene.
  • AAV8 adeno-associated virus 8
  • mice were subcutaneously administered a single 2 mg/kg of CFB siRNA agents or PBS.
  • Blood samples were collected at day 8, before dosing of siRNA, at day 18, day 25 and day32. Plasma samples were isolated and serum samples were collected for quantification protein level through ELISA was measured per manufacturer’s recommended protocol.
  • Cynomolgus monkeys (6 years old, 3 ⁇ 6 kg of weights, 3 monkeys in each group) were enrolled in the study. Each monkey was subcutaneously administered a single 6 mg/kg of CFB siRNA agents or PBS at day 1 (pre-dosing of siRNA) . After overnight fast, serum were drawn at day -14 (pre-dose) , day -7 (pre-dose) , 1 (pre-dose) , , 8, 15, 22, 29, 35, 43, 50, 57, 64, and 71; CFB protein remaining by Western blotting, the result is shown in table 13.
  • Cynomolgus monkeys (6 years old, 3 ⁇ 6 kg of weights, 3 monkeys in each group) were enrolled in the study. Each monkey was subcutaneously administered a single 3mg/kg of CFB siRNA agents or PBS at day 1 (pre-dosing of siRNA) . After overnight fast, serum were drawn at day -14 (pre-dose) , day -7 (pre-dose) , 1 (pre-dose) , all group Sample collection: Day -14, Day -7, 1, 8, 15, 22, 29, 35, 43, 50, and 57; CFB protein remaining by Western blotting (WB) , The result is shown in table 17.
  • WB Western blotting
  • Huh7 cells were trypsinized and adjusted to appropriate density seeded into 96-well plates.
  • Cells were transfected with the complexes of psiCHECK (TM) -2 Vector plasmid, blank vector pCNDNA 3.0, siRNAs or a control siRNA using Lipofectamine 2000 (Invitrogen-11668-019) at the next day of seeding following the protocol according to manufacturer’s recommendation.
  • the siRNAs were tested at different concentrations (10 nM and 1 nM, ) in triplicate.
  • psiCHECK (TM) Vector/blank vector pCDNA3.0/siRNAs/Lipofectamine 2000 mixture transfection.
  • No compound control well was defined as cells transfected with psiCHECK (TM) -2 Vector and blank vector pCNDNA 3.0 and without siRNA treatment; blank control was cell only wells.
  • Ratio of sample well (sample Renilla luminescence-background blank) / (sample Fireflyluminescence-background blank)
  • Ratio of no compound control well (control Renilla luminescence-background blank) / (control sample Fireflyluminescence-background blank)
  • Table 18 provides experimental results of in vitro studies using various CFB RNAi agents to inhibit CFB expression.
  • the duplex sequences used correspond to those shown in Table 2.

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Genetics & Genomics (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Molecular Biology (AREA)
  • Medicinal Chemistry (AREA)
  • Biomedical Technology (AREA)
  • Biochemistry (AREA)
  • Epidemiology (AREA)
  • Organic Chemistry (AREA)
  • Wood Science & Technology (AREA)
  • Biotechnology (AREA)
  • General Engineering & Computer Science (AREA)
  • Zoology (AREA)
  • Plant Pathology (AREA)
  • Microbiology (AREA)
  • Biophysics (AREA)
  • Physics & Mathematics (AREA)
  • Urology & Nephrology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)

Abstract

L'invention concerne des compositions et des procédés utiles pour réduire l'expression du gène CFB et pour le traitement de maladies et d'états associés à CFB. L'invention concerne des agents ARNdb du CFB, des agents polynucléotidiques antisens du CFB, des compositions comprenant des agents ARNdb du CFB, et des compositions comprenant des agents polynucléotidiques antisens du CFB qui peuvent être utilisés pour réduire l'expression de CFB chez des cellules et des sujets.
PCT/CN2024/102170 2023-06-28 2024-06-28 Compositions et procédés pour inhiber l'expression du facteur b du complément (cfb) Pending WO2025002299A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
CNPCT/CN2023/103142 2023-06-28
CN2023103142 2023-06-28

Publications (1)

Publication Number Publication Date
WO2025002299A1 true WO2025002299A1 (fr) 2025-01-02

Family

ID=93937679

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/CN2024/102170 Pending WO2025002299A1 (fr) 2023-06-28 2024-06-28 Compositions et procédés pour inhiber l'expression du facteur b du complément (cfb)

Country Status (2)

Country Link
TW (1) TW202500169A (fr)
WO (1) WO2025002299A1 (fr)

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2015038939A2 (fr) * 2013-09-13 2015-03-19 Isis Pharmaceuticals, Inc. Modulateurs du facteur b du complément
CN105814205A (zh) * 2013-12-12 2016-07-27 阿尔尼拉姆医药品有限公司 补体成分iRNA组合物及其使用方法
WO2023031359A1 (fr) * 2021-09-02 2023-03-09 Silence Therapeutics Gmbh Acides nucléiques pour inhiber l'expression du facteur b du complément (cfb) dans une cellule
WO2023045994A1 (fr) * 2021-09-23 2023-03-30 Shanghai Argo Biopharmaceutical Co., Ltd. Compositions et méthodes d'inhibition de l'expression de la protéine 3 (angptl3) de type angiopoïétine

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2015038939A2 (fr) * 2013-09-13 2015-03-19 Isis Pharmaceuticals, Inc. Modulateurs du facteur b du complément
CN105814205A (zh) * 2013-12-12 2016-07-27 阿尔尼拉姆医药品有限公司 补体成分iRNA组合物及其使用方法
WO2023031359A1 (fr) * 2021-09-02 2023-03-09 Silence Therapeutics Gmbh Acides nucléiques pour inhiber l'expression du facteur b du complément (cfb) dans une cellule
WO2023045994A1 (fr) * 2021-09-23 2023-03-30 Shanghai Argo Biopharmaceutical Co., Ltd. Compositions et méthodes d'inhibition de l'expression de la protéine 3 (angptl3) de type angiopoïétine

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
ADACHI,H. ET AL: "From antisens RNA to RNA modification: therapeutic potential of RNA-based technologies", BIOMEDICINES, vol. 9, 14 May 2021 (2021-05-14), XP055965840, DOI: 10.3390/biomedicines9050550 *
CHEN HUAN, LIJUN WEI: "Advances in research on non-viral delivery system for RNA drugs", PROGRESS IN PHARMACEUTICAL SCIENCES, CHINA PHARMACEUTICAL UNIVERSITY, CN, vol. 46, no. 11, 1 January 2022 (2022-01-01), CN , pages 839 - 847, XP093253568, ISSN: 1001-5094, DOI: 10.20053/j.issn1001-5094.2022.11.005 *
DATABASE NUCLEOTIDE 17 March 2023 (2023-03-17), ANONYMOUS: "Homo sapiens complement factor B (CFB), mRNA", XP093253567, Database accession no. NM_001710.6 *
JAFFE GLENN J, SAHNI JAYASHREE, FAUSER SASCHA, GEARY RICHARD S, SCHNEIDER EUGENE, MCCALEB MICHAEL: "Development of IONIS-FB-L to Treat Geographic Atrophy Associated with AMD", ARVO ANNUAL MEETING ABSTRACT, vol. 61, no. 7, 1 June 2020 (2020-06-01), pages 4305, XP093253566 *

Also Published As

Publication number Publication date
TW202500169A (zh) 2025-01-01

Similar Documents

Publication Publication Date Title
WO2023045994A1 (fr) Compositions et méthodes d'inhibition de l'expression de la protéine 3 (angptl3) de type angiopoïétine
EP4592390A1 (fr) Réactif d'arni spécifiquement modifié et composition
EP4442827A1 (fr) Composition et procédé d'inhibition de l'expression de la protéine du virus de l'hépatite b (hbv)
US20230092615A1 (en) Compositions and methods for inhibiting expressing of methylation-controlled j-protein (mcj)
WO2025077806A1 (fr) Compositions et procédés pour inhiber l'expression de la protéine tau associée aux microtubules (mapt)
WO2025108284A1 (fr) Compositions et procédés d'inhibition de l'expression de la sous-unité 9 du canal sodique voltage-dépendant (scn9a)
WO2024188164A1 (fr) Compositions et procédés d'inhibition de l'expression de l'effecteur b de type dffa induisant la mort cellulaire (cideb)
WO2025021007A1 (fr) Compositions et procédés pour inhiber l'expression du composant 3 du complément (c3)
WO2025002299A1 (fr) Compositions et procédés pour inhiber l'expression du facteur b du complément (cfb)
WO2025113470A1 (fr) Compositions et procédés d'inhibition de l'expression de la transthyrétine (ttr)
WO2023143483A1 (fr) Compositions et procédés pour inhiber l'expression de la protéine prékallikréine (pkk)
AU2024276138A1 (en) Compositions and methods for inhibiting expression of coagulation factor xi (fxi)
WO2024240058A1 (fr) Compositions et procédés pour inhiber l'expression du facteur xi de coagulation (fxi)
WO2025148896A1 (fr) Compositions et procédés d'inhibition de l'expression de sérine protéase transmembranaire 6 (tmprss6)
WO2025131019A1 (fr) Compositions et procédés pour inhiber l'expression de la sous-unité bêta e de l'inhibine (inhbe)
WO2025168022A1 (fr) Compositions et procédés d'inhibition de l'expression de huntingtine (htt)
WO2025077711A9 (fr) Compositions et procédés d'inhibition de l'expression de la protéine précurseur de l'amyloïde (app)
EP4630556A1 (fr) Compositions et méthodes pour inhiber l'expression du gène contenant le domaine de phospholipase de type patatine 3 (pnpla3)
WO2025016444A1 (fr) Compositions et procédés d'inhibition de l'expression de pd-l1
WO2024131916A1 (fr) Compositions et méthodes pour inhiber l'expression de 17bêta-hydroxystéroïde déshydrogénase de type 13 (hsd17b13)
CN121219417A (zh) 抑制凝血因子xi(fxi)表达的组合物和方法
OA21722A (en) Compositions and methods for inhibiting expression of Prekallikrein (PKK) protein.
HK40117932A (en) Composition and method for inhibiting expression of hepatitis b virus (hbv) protein
OA21711A (en) Composition and method for inhibiting expression of hepatitis B virus (HBV) protein.
JP2025542119A (ja) パタチン様ホスホリパーゼドメイン含有３（ｐｎｐｌａ３）の発現を阻害するための組成物及び方法

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 24830931

Country of ref document: EP

Kind code of ref document: A1