Products and compositions

Abstract

The present invention relates to products, compositions and uses thereof. In particular, the invention relates to nucleic acid products that modulate, in particular interfere with or inhibit the expression of the C5 gene. The product may be an oligomeric compound comprising at least a first linking nucleoside region having at least a first nucleoside base sequence at least partially complementary to at least a portion of RNA transcribed from C5 gene wherein the first nucleoside base sequence is selected from the group consisting of SEQ ID NO 1 to 250 or a portion thereof.

Description

[0001] Related Applications

[0002] This application claims the benefit and priority of U.S. Provisional Patent Application No. 63 / 352,503, filed on June 15, 2022, and U.S. Provisional Patent Application No. 63 / 408,318, filed on September 20, 2022. The entire contents of these two applications are incorporated herein by reference.

[0003] Sequence Listing

[0004] This application contains a sequence listing, which has been electronically submitted in XML format and is incorporated herein by reference in its entirety. The XML document was created on June 8, 2023, named 4690_0070I_SL, and is 4587 kilobytes in size.

[0005] Field

[0006] The present invention relates to products, compositions, and their uses. Specifically, the present invention relates to nucleic acid products that modulate, particularly interfere with or inhibit, the expression of the complement component C5 gene. Thus, embodiments of the present invention can provide methods, compounds, and compositions for reducing C5 mRNA and protein expression in animals. Such methods, compounds, and compositions can be used to treat, prevent, or improve complement system-related diseases, including C5-related diseases such as paroxysmal nocturnal hemoglobinuria (PNH), Alzheimer's disease, atherosclerosis, choroid plexus inflammation, generalized myasthenia gravis (gMG), amyotrophic lateral sclerosis (ALS), lupus nephritis (LN), central nervous system (CNS) diseases; age-related macular degeneration (AMD) and / or geographic atrophy (GA); uveitis and / or panuveitis; cold agglutinin disease, membranoproliferative glomerulonephritis (MPGN), Guillain-Barré syndrome, Shiga toxin-producing Escherichia coli hemolytic uremic syndrome (STEC-HUS), and organ transplantation-related autoimmune diseases.

[0007] Background

[0008] The complement system is part of the innate immune system. It is evolutionarily older than the adaptive immune system and is conserved in most taxa. Its functions include modifying potentially pathogenic microorganisms (a process called opsonization) and disrupting their infection, which is achieved by a large molecular assembly called the membrane attack complex (MAC). Once certain components of the complement system are activated, they promote the chemotaxis and activation of white blood cells.

[0009] Complement activation can be triggered by a variety of factors, all of which are related to the presence of microorganisms, but may also be related to components of the adaptive immune system, such as immunoglobulins (including immunoglobulin M). Three major pathways of complement activation have been identified, namely the classical pathway, the alternative pathway (or "alternative pathway"), and the lectin pathway.

[0010] Functionally, complement activation itself occurs at a low level (spontaneous cleavage of C3 to produce C3a and C3b), and is enhanced in the presence of microorganisms by converting inactive forms of enzymes (zymogens) to their active counterparts through an enzymatic cascade. The term "convertase", such as C3 convertase, is mainly a functional term and may refer to complexes with different structures. One C3 convertase is a complex of C3b and complement factor B (CFB, factor B). Once formed, C3 convertase can convert a large amount of C3 into its cleavage products C3a and C3b in a short time. A specific C3 convertase is a complex of C3b and factor B, which was initially discovered in the alternative pathway, but may also form in the context of the other two pathways. In the alternative pathway, factor B is also a component of C5 convertase, which is a complex that converts C5 (a downstream component of this pathway) into an active form. After C5 is formed, C5 convertase cleaves C5 into C5b and C5a, which is the starting event for the later steps of complement activation. Based on C5b, a membrane attack complex (MAC) is formed, which lyses the target membrane by constructing pores with C9 molecules.

[0011] Diseases

[0012] The complement system is usually triggered by patterns of surface binding sites. These binding sites may be components of microorganisms or pathogens, or may be previously formed antibodies that can bind to any target. In the latter case, the complement system plays a role in enhancing the adaptive immune system. Therefore, if the antibody is an autoantibody, the complement system will exacerbate the adverse autoimmune response. Interfering with the complement system in this case is a means of treating or improving autoimmune diseases. Since the complement system, more specifically the C1 of the classical pathway, recognizes the constant region of the antibody, interfering with the complement system opens up a way to interfere with autoimmune diseases without particular limitations. Experience shows that autoimmune diseases affect the skin, joints, and kidneys more frequently than other organs.

[0013] On the other hand, in the absence of autoantibodies, complement dysfunction may also trigger diseases. In addition, in this case, due to the general mechanism of the complement system, specific diseases can be treated without particular limitations by complement system inhibitors (more specifically, by complement component C5 inhibitors).

[0014] Treatment

[0015] Eculizumab is a humanized monoclonal antibody against C5 and has been approved for the treatment of PNH (paroxysmal nocturnal hemoglobinuria). Its production uses a murine cell line. Patients sensitive to murine proteins must not use eculizumab. The treatment cost of eculizumab is very expensive, and the cost for each patient per year may exceed 600,000 euros.

[0016] Therefore, there is still a need for therapies for treating complement-related diseases (including complement component C5-related diseases). To this end, we aim to provide compounds, methods, and pharmaceutical compositions for treating such diseases.

[0017] Double-stranded RNA (dsRNA) capable of complementary binding to expressed mRNA has been shown to block gene expression through a mechanism called RNA interference (RNAi) (Fire et al., 1998, Nature. February 19, 1998; 391(6669):806-11 and Elbashir et al., 2001, Nature. May 24, 2001; 411(6836):494-8). Short dsRNAs direct gene-specific, post-transcriptional silencing in many organisms, including vertebrates, and have become useful tools for studying gene function. RNAi is mediated by the RNA-induced silencing complex (RISC), a sequence-specific, multi-component nuclease that destroys messenger RNAs homologous to the silencing trigger loaded into the RISC complex. Interfering RNAs (iRNAs), such as siRNAs, antisense RNAs, and microRNAs, are all oligonucleotides that prevent protein formation through gene silencing, that is, they inhibit the gene translation of proteins by degrading mRNA molecules. Gene silencers are becoming increasingly important in medical treatment applications.

[0018] According to the article by Watts and Corey in the Journal of Pathology (2012; Vol. 226, pp. 365-379), there are algorithms available for designing nucleic acid silencing triggers, but all of these algorithms have serious limitations. Various experimental methods may be required to identify effective siRNAs because the algorithms do not take into account factors such as the tertiary structure of the target mRNA or the involvement of RNA-binding proteins. Therefore, finding effective nucleic acid silencing triggers with minimal off-target effects is a complex process. For the drug development of these highly charged molecules, it must be possible to synthesize them economically, distribute them to the target tissues, enter the cells, and function within an acceptable toxicity range. Therefore, the aim is to provide compounds, methods, and pharmaceutical compositions for treating the C5-related diseases described herein, which comprise oligomeric compounds that regulate (especially inhibit) gene expression through RNAi.

[0019] Overview

[0020] The present invention solves the problem of providing compounds and treatment methods with the potential to effectively reduce the impact of C5-related diseases as described above.

[0021] According to a first aspect, the present invention relates to an oligomeric compound capable of inhibiting the expression of complement component C5, wherein the compound comprises at least a first linked nucleoside region, and the at least first nucleoside base sequence is at least partially complementary to at least a part of the RNA transcribed from the C5 gene, and the first nucleoside base sequence is selected from the sequences in Table 1a (SEQ ID NO: 1 to 250) or a part thereof, and the part preferably has a length of at least 18 nucleosides.

[0022] A particularly preferred embodiment according to the first aspect of the present invention relates to an optimized hairpin RNA (referred to as mxRNA); for more details, please refer to the embodiments and their discussions below.

[0023] Furthermore, as further disclosed below, the present invention also relates to double-stranded RNA (dsRNA). Different from mxRNA, dsRNA lacks the loop connecting the antisense and sense parts and thus comprises two strands. The two strands are not covalently linked but form a double-stranded region where base pairing occurs.

[0024] According to a second aspect, the present invention relates to a nucleic acid construct comprising at least:

[0025] (a) a first nucleic acid part that is at least partially complementary to at least a first part of the RNA transcribed from the C5 gene;

[0026] (b) a second nucleic acid part that is at least partially complementary to at least a second part of the RNA transcribed from the C5 gene, and the second part is different from the first part;

[0027] (c) a third nucleic acid part that is at least partially complementary to the first nucleic acid part of (a) and thus forms a first nucleic acid duplex region therewith;

[0028] (d) a fourth nucleic acid part that is at least partially complementary to the second nucleic acid part of (b) and thus forms a second nucleic acid duplex region therewith.

[0029] Preferred and / or exemplary features of the constructs according to the second aspect of the present invention are as follows:

[0030] 1) They contain multiple (two or more) at least partially double-stranded RNA interference-triggering factors that are mainly linked together through complementary (Watson-Crick) interactions to form a single nanostructure;

[0031] 2) Optionally, other covalent linkages (e.g.,) can be used to construct the construct and / or add various ligands (e.g., delivery / targeting moieties such as GalNAc and / or other carbohydrates, cholesterol, peptides, or small molecules, optionally linked via a linker);

[0032] 3) The constructs of the present invention mainly contain chemically modified nucleotides (e.g., 2'F, 2'OMe, LNO, PNA, MOE, BNA, PMO, phosphorothioate, dithiophosphate, etc.), mainly (but not limited to) to increase resistance to nucleases;

[0033] 4) The construct contains "labile" components (e.g., chemical linkers, unmodified nucleotides, etc.), which cause the construct to decompose when exposed to certain biological environments (e.g., exposure to extracellular and / or intracellular fluids); specific examples can be (but are not limited to): a) the oligonucleotide backbone is cleaved by nucleases at the site of unmodified nucleotides; b) chemical bonds are broken due to changes in pH (e.g., in endosomes);

[0034] 5) After dissociation upon exposure to said certain biological environments, the active ingredient (e.g., the agent capable of triggering RNA interference of at least a part of the double strand) is released to regulate (upregulate or downregulate, preferably downregulate) the expression of target genes in cells / organisms;

[0035] 6) The construct can be used to regulate, preferably downregulate or silence gene expression, study gene function, or treat various diseases related to downregulation of target genes.

[0036] According to a third aspect, the present invention relates to a composition comprising an oligomeric compound according to the first aspect and / or a nucleic acid construct according to the second aspect, and a physiologically acceptable excipient.

[0037] According to a fourth aspect, the present invention relates to a pharmaceutical composition comprising an oligomeric compound according to the first aspect and / or a nucleic acid construct according to the second aspect.

[0038] According to a fifth aspect, the present invention relates to an oligomeric compound according to the first aspect and / or a nucleic acid construct according to the second aspect for use in human or veterinary medicine or therapy.

[0039] According to a sixth aspect, the present invention relates to an oligomeric compound according to the first aspect and / or a nucleic acid construct according to the second aspect for use in a method of treating a disease or disorder.

[0040] According to a seventh aspect, the present invention relates to a method of treating a disease or disorder, comprising administering to an individual in need of treatment an oligomeric compound according to the first aspect and / or a nucleic acid construct according to the second aspect.

[0041] According to an eighth aspect, the present invention relates to the use of an oligomeric compound according to the first or second aspect for research as a gene function analysis tool.

[0042] According to a ninth aspect, the present invention relates to the use of an oligomeric compound according to the first or second aspect in the manufacture of a medicament for treating a disease or disorder.

[0043] Effects achieved by the oligomeric compounds of the present invention

[0044] Due to the use of the oligomeric compounds of the present invention, as shown in the examples disclosed herein, a significant reduction in the expression of complement component C5 in vitro and in vivo was achieved. The most inhibitory compound surprisingly produced up to a 70% to 75% reduction in C5 mRNA in vitro. In addition, a large number of compounds were also able to produce a 60% to 70% reduction in C5 mRNA in vitro. The significant in vitro activity of these compounds was also confirmed in the in vivo studies disclosed herein. Therefore, the oligomeric compounds of the present invention are suitable candidates for treating C5-related diseases.

[0045] Furthermore, surprisingly, in certain embodiments, the use of an oligomeric compound according to the present invention to inhibit the expression of C5 achieves the above effects, and the oligomeric compound is in the form of an shRNA construct, which has a shorter length (e.g., 33 nucleotides) compared to a conventional shRNA molecule with a longer length. This can, for example, make the synthesis of the shRNA molecule more efficient because fewer units are required.

[0046] For certain oligomeric compounds of the present invention, which are in the form of shRNA constructs for inhibiting the expression of C5, it was surprisingly found that the above effects can be achieved by using a short sense strand in the shRNA, and the length of the short sense strand is preferably 14 nucleotides, which is shorter than the length of the sense strand in a conventional shRNA molecule.

[0047] Due to the successful C5 knockdown by the compounds of the present invention in the mxRNA form, they also have the potential to act in the same manner as the muRNA constructs disclosed herein. This is especially the case without being bound by a particular theory, assuming that the active species are the same. Brief description of the drawings

[0049] Figure 1 Shows the single-dose curves of certain C5 mxRNA compounds of the present invention and their activity in inhibiting C5 gene expression (preliminary screening).

[0050] Figure 2 Shows the dose curves of 25 C5 mxRNA compounds and their activity in inhibiting C5 gene expression (secondary screening).

[0051] Figure 3 Dose curve prepared in vivo for the C5 mxRNA lead compound.

[0052] Figure 4 Shows the research plan and research information related to the C5-targeted mxRNA study, which provides clues for the candidate dose and duration response study in the humanized liver-uPA-SCID mouse (PXB) model.

[0053] Figure 5 Shows the results of the dose and duration response study of the C5-targeted mxRNA construct in the humanized liver-uPA-SCID mouse (PXB).

[0054] Figure 6 Shows the research plan and research information related to the study of the duration effect of mxRNA targeting human complement C5 in the non-GLP model of humanized liver-uPA-SCID mice.

[0055] Figure 7 Shows the results of the study of the duration effect of mxRNA targeting human complement C5 in the non-GLP model of humanized liver-uPA-SCID mice.

[0056] Detailed description and examples

[0057] The following further embodiments (items) of the present invention are described only by way of example. These examples represent the best ways currently known to the applicant to put the present invention into practice, although they are not the only ways to achieve this goal.

[0058] It should be understood that the benefits and advantages described herein may relate to one embodiment or multiple embodiments. The embodiments are not limited to those that solve any or all of the problems described, nor are they limited to those that have any or all of the benefits and advantages described. The embodiments marked as "preferred", "preferably" or "preferably" are not intended to limit the scope of the claims, but to show alternative embodiments of the present invention.

[0059] The features of different aspects and embodiments of the present invention can be appropriately combined, which is obvious to those skilled in the art and can be combined with any aspect of the present invention.

[0060] Definition

[0061] The following definitions run throughout the present invention. In many cases, in addition to the definitions themselves, these definitions also provide a non-exhaustive list of possible implementations, which are equivalent to the preferred embodiments.

[0062] Unless otherwise defined, the terms used in analytical chemistry, synthetic organic chemistry, and medicinal and pharmaceutical chemistry, as well as their procedures and techniques described herein, are well known and commonly used in the art. Standard techniques are available for chemical synthesis and chemical analysis. Certain such techniques and procedures can be found, for example, in "Carbohydrate Modifications in Antisense Research", edited by Sangvi and Cook, American Chemical Society, Washington, D.C., 1994; "Remington: The Science and Practice of Pharmacy", Mack Publishing Co., Easton, Pa., 21st Edition, 2005; and "Antisense Drug Technology, Principles, Strategies, and Applications", edited by Stanley T. Crooke, CRC Press, Boca Raton, Fla.; and Sambrook et al., "Molecular Cloning, A Laboratory Manual", 2nd Edition, Cold Spring Harbor Laboratory Press, 1989, which are hereby incorporated by reference herein for any purpose. To the extent permitted, all patents, applications, published applications, and other publications and other data mentioned in this disclosure are incorporated herein by reference in their entirety.

[0063] Unless otherwise noted, the following terms have the following meanings:

[0064] As used herein, "excipient" refers to any compound or mixture of compounds added to the compositions provided herein that is suitable for delivering an oligomeric compound.

[0065] "Nucleoside" as used herein refers to a compound that includes a nucleobase moiety and a sugar moiety. Nucleosides include, but are not limited to, naturally occurring nucleosides (such as those found in DNA and RNA) and modified nucleosides. A nucleoside can be linked to a phosphate moiety, and a phosphate-linked nucleoside is also referred to as a "nucleotide". The structural features and / or length of the oligomeric compounds or nucleic acid constructs disclosed herein are expressed in terms of "nucleosides" or "nucleotides".

[0066] "Chemical modification" or "chemically modified" as used herein refers to a chemical difference of a compound compared to its naturally occurring counterpart. Chemical modifications of oligonucleotides include nucleoside modifications (including sugar moiety modifications and nucleobase modifications) and internucleoside bond modifications. For oligonucleotides, chemical modifications do not include differences in only the nucleobase sequence.

[0067] "Furanosyl" as used herein refers to the structure of a five-membered ring containing four carbon atoms and one oxygen atom.

[0068] As used herein, "naturally occurring sugar moiety" refers to ribofuranosyl in naturally occurring RNA or deoxyribofuranosyl in naturally occurring DNA. The "naturally occurring sugar moiety" as referred to herein is also referred to as "unmodified sugar moiety". Specifically, such "naturally occurring sugar moiety" or "unmodified sugar moiety" as referred to herein has -H (DNA sugar moiety) or -OH (RNA sugar moiety) at the 2'-position of the sugar moiety, especially -H (DNA sugar moiety) at the 2'-position of the sugar moiety.

[0069] As used herein, "sugar moiety" refers to the naturally occurring sugar moiety or modified sugar moiety of a nucleoside. The "modified sugar moiety" as used herein refers to a substituted sugar moiety or sugar surrogate.

[0070] As used herein, "substituted sugar moiety" refers to a furanosyl that has been substituted. Substituted sugar moieties include, but are not limited to, furanosyls containing substituents at the 2'-position, 3'-position, 5'-position, and / or 4'-position. Certain substituted sugar moieties are bicyclic sugar moieties.

[0071] As used herein, "2'-substituted sugar moiety" refers to a furanosyl containing a substituent other than H or OH at the 2'-position. Unless otherwise specified, the 2'-substituted sugar moiety is not a bicyclic sugar moiety (i.e., the 2'-substituent of the 2'-substituted sugar moiety does not form a bridge with another atom of the furanosyl ring).

[0072] As used herein, "MOE" represents -OCH2CH2OCH3.

[0073] As used herein, "2'-F nucleoside" refers to a nucleoside containing a sugar with fluorine at the 2'-position. Unless otherwise specified, the fluorine in the 2'-F nucleoside is at the ribose position (substituting the OH of natural ribose). A uniformly modified 2'-fluorinated (ribose) oligonucleotide duplex hybridizing to an RNA strand is not an RNase H substrate, while the analogue retains RNase H activity.

[0074] As used herein, the term "sugar surrogate" refers to a structure that does not contain a furanosyl and is capable of substituting the natural sugar moiety of a nucleoside such that the resulting nucleoside subunit can be linked together and / or linked to other nucleosides to form an oligomeric compound capable of hybridizing to a complementary oligomeric compound. Such structures include rings containing a different number of atoms than a furanosyl (e.g., 4, 6, or 7-membered rings); substitution of the oxygen of the furanosyl with a non-oxygen atom (e.g., carbon, sulfur, or nitrogen); or a change in the number of atoms and oxygen substitution. Such structures may also contain substitutions corresponding to those described for substituted sugar moieties (e.g., a 6-membered carbon ring bicyclic sugar surrogate may optionally contain additional substituents). Sugar surrogates also include more complex sugar substitutions (e.g., the acyclic system of peptide nucleic acid). Sugar surrogates include, but are not limited to, morpholine, cyclohexenyl, and cyclohexitol.

[0075] As used herein, the term "bicyclic sugar moiety" refers to a modified sugar moiety containing a 4- to 7-membered ring (including but not limited to furanosyl), which contains a bridge connecting two atoms of the 4- to 7-membered ring to form a second ring, thereby producing a bicyclic structure. In certain embodiments, the 4- to 7-membered ring is a sugar ring. In certain embodiments, the 4- to 7-membered ring is furanosyl. In certain such embodiments, the bridge connects the 2'-carbon and the 4'-carbon of the furanosyl.

[0076] As used herein, the term "nucleotide" refers to a nucleoside that further includes a phosphate linking group. As used herein, "linked nucleosides" may or may not be linked by a phosphate bond, and thus include but are not limited to "linked nucleotides". As used herein, "linked nucleosides" are nucleosides linked in a continuous sequence (i.e., there are no additional nucleosides between the linked nucleosides).

[0077] As used herein, the term "nucleobase" refers to a group of atoms that can be linked to a sugar moiety to form a nucleoside capable of being incorporated into an oligonucleotide, and wherein the group of atoms is capable of binding to a complementary naturally occurring nucleobase of another oligonucleotide or nucleic acid, more specifically by hydrogen bonding. Nucleobases can be naturally occurring or modified.

[0078] As used herein, the term "unmodified nucleobase" or "naturally occurring nucleobase" refers to the naturally occurring heterocyclic nucleobases of RNA or DNA: the purine bases adenine (A) and guanine (G), and the pyrimidine bases thymine (T), cytosine (C) (including 5-methyl C) and uracil (U).

[0079] As used herein, a "modified nucleobase" refers to any non-naturally occurring nucleobase.

[0080] As used herein, a "modified nucleoside" refers to a nucleoside that contains at least one chemical modification compared to a naturally occurring RNA or DNA nucleoside. Modified nucleosides can contain a modified sugar moiety and / or a modified nucleobase.

[0081] As used herein, "bicyclic nucleoside" or "BNA" refers to a nucleoside that contains a bicyclic sugar moiety.

[0082] As used herein, "locked nucleic acid nucleoside" or "LNA" refers to a nucleoside that contains a bicyclic sugar moiety that contains a 4'-CH2-O-2' bridge.

[0083] As used herein, a "2'-substituted nucleoside" refers to a nucleoside that contains a substituent other than H or OH at the 2'-position of the sugar moiety. Unless otherwise specified, 2'-substituted nucleosides are not bicyclic nucleosides.

[0084] As used herein, "deoxynucleoside" refers to a nucleoside containing a 2'-H furanosyl sugar moiety, such as found in naturally occurring deoxyribonucleosides (DNA). In certain embodiments, the 2'-deoxynucleoside may contain a modified nucleobase or may contain an RNA nucleobase (e.g., uracil).

[0085] As used herein, "oligonucleotide" refers to a compound containing multiple linked nucleosides. In certain embodiments, the oligonucleotide contains one or more unmodified ribonucleosides (RNA) and / or unmodified deoxyribonucleosides (DNA) and / or one or more modified nucleosides.

[0086] As used herein, "modified oligonucleotide" refers to an oligonucleotide containing at least one modified nucleoside and / or at least one modified internucleoside bond.

[0087] Preferred modified internucleoside bonds are those that have higher stability compared to the naturally occurring phosphodiester. "Stability" particularly refers to stability against hydrolysis (including enzyme-catalyzed hydrolysis, enzymes including exonucleases and endonucleases).

[0088] Preferred positions for such modified internucleoside bonds include the termini and hairpin loops of the single-stranded oligomeric compounds of the present invention. For example, modify the internucleoside bonds between the first and second nucleosides and between the second and third nucleosides counted from the 5'-terminus, and / or modify the internucleoside bonds between the first and second nucleosides and between the second and third nucleosides counted from the 3'-terminus. In addition, the bond between the terminal nucleoside linked to the 3'-terminus and a ligand (e.g., GalNAc) can be modified.

[0089] As described above, the preferred positions are in the hairpin loops of the single-stranded oligomeric compounds. Specifically, all the bonds, all but one bond, or most of the bonds in the hairpin loop are modified. As used herein, "bonds in the hairpin loop" refers to the bonds between nucleosides that do not participate in base pairing. For example, in a hairpin loop composed of five nucleosides, there are four bonds between nucleosides that do not participate in base pairing. Preferably, the term "bonds in the hairpin loop" also extends to the bonds connecting the stem to the loop, i.e., the bonds connecting base-paired nucleosides to non-base-paired nucleosides. Generally, according to the present invention, there are two such positions in the hairpin and mxRNA.

[0090] Most preferably, the modified internucleoside bonds are located at both ends and in the hairpin loop.

[0091] As used herein, "linking" or "linking group" refers to a group of atoms that connects two or more other groups of atoms together.

[0092] As used herein, "internucleoside bond" refers to the covalent bond between adjacent nucleosides in an oligonucleotide.

[0093] As used herein, the "naturally occurring internucleoside bond" refers to a 3' to 5' phosphodiester bond.

[0094] As used herein, the "modified internucleoside bond" refers to any internucleoside bond other than the naturally occurring internucleoside bond. Specifically, the "modified internucleoside bond" as referred to herein may include a modified phosphorus linking group, such as a phosphorothioate or dithiophosphonate internucleoside bond.

[0095] As used herein, the "terminal internucleoside bond" refers to the bond between the last two nucleosides of an oligonucleotide or a defined region thereof.

[0096] As used herein, the "phosphorus linking group" refers to a linking group containing a phosphorus atom, which may include naturally occurring phosphorus linking groups, such as those present in naturally occurring RNA or DNA, such as phosphodiester linking groups, or modified phosphorus linking groups not normally present in naturally occurring RNA or DNA, such as phosphorothioate or dithiophosphonate linking groups. Thus, the phosphorus linking group may include, but is not limited to, phosphodiester, phosphorothioate, dithiophosphonate, phosphonate, methylphosphonate, phosphoramide, phosphorothioamide, thioalkylphosphonate, phosphotriester, thioalkyl phosphotriester, and boranophosphonate.

[0097] As used herein, the "internucleoside phosphorus linking group" refers to a phosphorus linking group that directly links two nucleosides.

[0098] As used herein, an "oligomeric compound" refers to a polymeric structure comprising two or more substructures. In certain embodiments, the oligomeric compound comprises an oligonucleotide, such as a modified oligonucleotide. In certain embodiments, the oligomeric compound further comprises one or more conjugate groups and / or terminal groups and / or ligands. In certain embodiments, the oligomeric compound consists of oligonucleotides. In certain embodiments, the oligomeric compound comprises a backbone of one or more linked monomeric sugar moieties, wherein each linked monomeric sugar moiety is directly or indirectly linked to a heterocyclic base moiety. In certain embodiments, the oligomeric compound may also include monomeric sugar moieties not linked to the heterocyclic base moiety, thereby providing abasic sites. The oligomeric compound may be defined solely based on the nucleoside base sequence, i.e., by specifying the sequence of A, G, C, U (or T). In this case, the structure of the sugar-phosphate backbone is not particularly limited and may or may not include modified sugars and / or modified phosphates. On the other hand, the oligomeric compound may be more comprehensively defined, i.e., by specifying not only the nucleoside base sequence but also the structure of the backbone, particularly the modification status of the sugar (unmodified, 2'-OMe modified, 2'-F modified, etc.) and / or the phosphate. mxRNA is a non-limiting example of an oligomeric compound.

[0099] As used herein, "nucleic acid construct" or "construct" refers to an assembly of two or more oligomeric compounds (e.g., four oligomeric compounds). The oligomeric compounds can be linked to each other by covalent bonds (e.g., phosphodiester bonds, such as those found in naturally occurring nucleic acids or modified versions thereof as disclosed herein) or by non-covalent bonds (e.g., hydrogen bonds, preferably hydrogen bonds between nucleobases, such as Watson-Crick base pairing). In certain embodiments, preferably, the construct comprises four oligomeric compounds, wherein two oligomeric compounds are covalently linked, thereby producing two nucleic acid strands that bind to each other by hydrogen bonds. The complementarity between the strands can be throughout the entire strands, but it does not necessarily have to be so. Specifically, an exemplary embodiment provides an antisense strand targeting a first region of C5 mRNA, which is covalently linked to the sense strand of another C5-targeting double-stranded RNA molecule, and the antisense strand of the C5 mRNA-targeting double-stranded RNA molecule is covalently linked to the sense strand of another C5 mRNA-targeting double-stranded RNA molecule. Since the antisense and sense strands of the parental single-targeting RNA molecule do not need to have the same length and preferably do not have the same length, with the antisense portion being longer than the sense portion, the preferred constructs of the present invention comprise a central region where the 3' regions of the antisense portions of the parental single-targeting RNA molecules face each other. Base pairing usually does not occur or only partially occurs in this region, but complete complementarity is not excluded. Otherwise, when the antisense and sense portions of each parental RNA molecule face each other; there is complementarity, preferably complete complementarity or 1 or 2 mismatches. MuRNA is a non-limiting example of a nucleic acid construct.

[0100] The term "strand" has its established meaning in the art and refers to a plurality of linked nucleosides. The linker is not particularly limited, but includes phosphodiesters and their variants as disclosed herein. A strand can also be regarded as a plurality of linked nucleotides, in which case the linker will be a covalent bond.

[0101] As used herein, "terminal group" refers to one or more atoms attached to one or both of the 3'-end or 5'-end of an oligonucleotide, also referred to as "terminus". In certain embodiments, the terminal group includes one or more terminal group nucleosides, and a "terminal nucleoside" is only one nucleotide at the corresponding terminus (5'-end or 3'-end).

[0102] As used herein, "conjugate" or "conjugate group" refers to an atom or group of atoms that binds to an oligonucleotide or oligomeric compound. In certain embodiments, the conjugate group links a ligand to a modified oligonucleotide or oligomeric compound. Generally, the conjugate group can modify one or more properties of the compound to which it is linked, including but not limited to pharmacodynamics, pharmacokinetics, binding, absorption, cellular distribution, cellular uptake, and charge and / or clearance properties.

[0103] As used herein, in the context of a conjugate group, a "conjugate linker" or "linker" refers to a portion of the conjugate group that includes any atom or group of atoms and covalently attaches an oligonucleotide to another portion of the conjugate group. In certain embodiments, the point of attachment on the oligomeric compound is the 3'-oxygen atom of the 3'-hydroxyl group of the 3'-terminal nucleoside of the oligonucleotide. In certain embodiments, the point of attachment on the oligomeric compound is the 5'-oxygen atom of the 5'-hydroxyl group of the 5'-terminal nucleoside of the oligonucleotide. In certain embodiments, the bond formed to attach to the oligomeric compound is a cleavable bond. In certain such embodiments, such cleavable bond constitutes all or part of a cleavable moiety.

[0104] In certain embodiments, the conjugate group comprises a cleavable moiety (e.g., a cleavable bond or a cleavable nucleoside) and a ligand moiety that can comprise one or more ligands, such as a carbohydrate cluster moiety, such as N-acetylgalactosamine, also referred to as "GalNAc", cluster moiety. In certain embodiments, the carbohydrate cluster moiety is identified by the number and identity of the ligands. For example, in certain embodiments, the carbohydrate cluster moiety comprises 2 GalNAc groups. For example, in certain embodiments, the carbohydrate cluster moiety comprises 3 GalNAc groups, which are particularly preferred. In certain embodiments, the carbohydrate cluster moiety comprises 4 GalNAc groups. Such ligand moieties are attached to the oligomeric compound via a cleavable moiety (e.g., a cleavable bond or a cleavable nucleoside). The ligands can be arranged in a linear or branched structure, such as a bi-antennary or tri-antennary structure. Preferred carbohydrate clusters have the following formula:

[0105]

[0106] , wherein one, two, or three of the phosphodiester bonds in the structural formula can also be replaced by phosphorothioate bonds.

[0107] As used herein, a "cleavable moiety" refers to a bond or group that is capable of cleaving under physiological conditions. In certain embodiments, the cleavable moiety cleaves within a cell or subcellular compartment (e.g., an endosome or a lysosome). In certain embodiments, the cleavable moiety is cleaved by an endogenous enzyme (e.g., a nuclease). In certain embodiments, the cleavable moiety comprises a group of atoms having one, two, three, four, or more than four cleavable bonds. In certain embodiments, the cleavable moiety is a phosphodiester bond.

[0108] As used herein, a "cleavable bond" refers to any chemical bond that is capable of being broken.

[0109] As used herein, a "carbohydrate cluster" refers to a compound having one or more carbohydrate residues attached to a linking group.

[0110] As used herein, "modified carbohydrate" refers to any carbohydrate having one or more chemical modifications relative to a naturally occurring carbohydrate.

[0111] As used herein, "carbohydrate derivative" refers to any compound that can be synthesized using a carbohydrate as a starting material or intermediate.

[0112] As used herein, "carbohydrate" refers to a naturally occurring carbohydrate, a modified carbohydrate, or a carbohydrate derivative. Carbohydrates are biomolecules that include carbon (C), hydrogen (H), and oxygen (O) atoms. Carbohydrates can include monosaccharides, disaccharides, trisaccharides, tetrasaccharides, oligosaccharides, or polysaccharides, such as one or more galactose moieties, one or more lactose moieties, one or more N-acetylgalactosamine moieties, and / or one or more mannose moieties. A particularly preferred carbohydrate is N-acetylgalactosamine.

[0113] As used herein, "strand" refers to an oligomeric compound that contains linked nucleosides.

[0114] As used herein, "single-stranded" or "single-strandedness" refers to an oligomeric compound composed of linked nucleosides that are linked in a continuous sequence without interruption. Such single strands may include regions that are sufficiently self-complementary to be able to form stable self-double strands in a hairpin structure.

[0115] As used herein, "hairpin" refers to a single-stranded oligomeric compound that includes a duplex formed by base pairing between self-complementary and oppositely directed sequences in the strand.

[0116] As used herein, "hairpin loop" refers to an unpaired loop of linked nucleosides in a hairpin structure formed by hybridization of self-complementary sequences. The resulting structure looks like a loop or a U shape.

[0117] Specifically, short hairpin RNA (also referred to as shRNA) contains a duplex region and a loop that connects the regions forming the duplex. The ends of the duplex region do not carry a loop and can be blunt-ended or carry a (a) 3' and / or a (a) 5' overhang. A blunt-ended construct is preferred. The term "shRNA" is more general than "mxRNA" (defined below) and can include compounds in which the loop is not or not entirely formed by the antisense strand. Specifically, shRNA includes an antisense strand (also referred to as the guide strand) that is complementary to a target RNA region and a sense strand (i.e., the passenger strand) that is substantially complementary to the antisense strand. More specifically, the antisense strand and the sense strand in shRNA are directly linked, for example, by a phosphodiester or phosphorothioate linkage, or by a third moiety forming a loop that links the nucleosides, which means that the 3' end of the antisense strand is covalently linked to the 5' end of the sense strand through several other groups. Such direct linkage does not include gaps or nicks.

[0118] As used herein, "directionality" refers to the end-to-end chemical orientation of an oligonucleotide, which is based on the chemical convention of numbering the carbon atoms of the sugar moiety, meaning that there will be a 5' end defined by the 5' carbon of the sugar moiety and a 3' end defined by the 3' carbon of the sugar moiety. In a double-stranded or duplex oligonucleotide, the strands run in opposite 5' to 3' directions to allow base pairing between them.

[0119] As used herein, "duplex" or the abbreviation "dup" refers to two or more complementary strand regions or strands of an oligonucleotide that hybridize together through non-covalent sequence-specific interactions. Most commonly, the hybridization in a duplex is between the nucleobases adenine

[0120] (A) and thymine (T), and / or between (A) adenine and uracil (U), and / or between guanine (G) and cytosine (C). A duplex can be part of a single-stranded structure where self-complementarity results in hybridization, or as a result of hybridization between the strands in a double-stranded construct.

[0121] As used herein, "double-stranded" or "duplex" refers to a pair of oligomeric compounds that hybridize to each other. In certain embodiments, the double-stranded oligomeric compound comprises a first and a second oligomeric compound.

[0122] As used herein, "expression" refers to the process by which a gene ultimately gives rise to a protein. Expression includes, but is not limited to, transcription, post-transcriptional modification (e.g., splicing, polyadenylation, addition of a 5'-cap), and translation.

[0123] As used herein, "transcription" or "transcribing" refers to the first of several steps in the expression of a DNA gene, in which the target sequence of DNA is copied into RNA (especially mRNA) by the enzyme RNA polymerase. During transcription, the DNA sequence is read by RNA polymerase, resulting in the production of a complementary antiparallel RNA sequence called the primary transcript.

[0124] As used herein, "target sequence" refers to the sequence to which an oligomeric compound is designed to hybridize to produce the desired C5 expression activity. The oligonucleotide has sufficient complementarity to its target sequence to allow hybridization under physiological conditions.

[0125] As used herein, "nucleobase complementarity" or "complementarity", when referring to nucleobases, means a nucleobase that is capable of base pairing with another nucleobase. For example, in DNA, adenine (A) is complementary to thymine (T). For example, in RNA, adenine (A) is complementary to uracil (U). In DNA and RNA, guanine (G) is complementary to cytosine (C). In certain embodiments, a complementary nucleobase refers to a nucleobase of an oligomeric compound that is capable of base pairing with the nucleobases of its target sequence. For example, if the nucleobase at a certain position of an oligomeric compound can form a hydrogen bond with the nucleobase at a certain position of the target sequence, the hydrogen bond position between the oligomeric compound and the target sequence is considered to be complementary at that nucleobase pair. Nucleobases containing certain modifications can retain the ability to pair with the corresponding nucleobases and thus can still achieve nucleobase complementarity.

[0126] As used herein, "non-complementary" with respect to nucleobases means a pair of nucleobases that do not form a hydrogen bond with each other.

[0127] As used herein, "complementary" with respect to an oligomeric compound (e.g., linked nucleosides, oligonucleotides) means the ability of such an oligomeric compound or a region thereof to hybridize to a target sequence or a region of the oligomeric compound itself through nucleobase complementarity.

[0128] A complementary oligomeric compound does not need to have nucleobase complementarity at every nucleoside. Instead, some mismatches can be tolerated. In certain embodiments, the complementary oligomeric compound or region is complementary at 70% of the nucleobases (70% complementary). In certain embodiments, the complementary oligomeric compound or region is 80% complementary. In certain embodiments, the complementary oligomeric compound or region is 90% complementary. In certain embodiments, the complementary oligomeric compound or region is at least 95% complementary. In certain embodiments, the complementary oligomeric compound or region is 100% complementary. As used herein, "self-complementarity" with respect to an oligomeric compound means that the compound can fold upon itself such that a double strand is generated due to the hybridization of the nucleobases in the internal complementary strand regions. Depending on the tightness and / or length of the strand regions, the compound can form hairpin loops, junctions, bulges, or internal loops.

[0129] As used herein, "mismatch" means that when an oligomeric compound is aligned with a target sequence and / or the self-complementary region of the oligomeric compound, the nucleobase of the oligomeric compound cannot base pair with the nucleobase at the corresponding position of the target sequence, or when the oligomeric compound hybridizes due to self-complementarity, the nucleobases at the corresponding positions of the oligomeric compound itself cannot pair.

[0130] As used herein, "hybridization" refers to the pairing of complementary oligomeric compounds (e.g., an oligomeric compound and its target sequence). While not limited to a particular mechanism, the most common pairing mechanism involves hydrogen bonding between complementary nucleobases, which can be Watson-Crick, Hoogsteen, or reverse Hoogsteen hydrogen bonds.

[0131] As used herein, "specific hybridization" refers to the ability of an oligomeric compound to hybridize to one nucleic acid site with a greater affinity than to another nucleic acid site.

[0132] As used herein, "fully complementary" refers to an oligomeric compound or a region thereof, which means that each nucleobase of the oligomeric compound or a region thereof is capable of pairing with a nucleobase of a complementary nucleic acid target sequence or a self-complementary region of the oligomeric compound. Thus, a fully complementary oligomeric compound or a region thereof does not contain mismatched or unhybridized nucleobases relative to its target sequence or the self-complementary region of the oligomeric compound.

[0133] As used herein, "complementary rate" refers to the percentage of nucleobases in an oligomeric compound that are complementary to an equally long portion of a target nucleic acid. The complementary rate is calculated by dividing the number of nucleobases in the oligomeric compound that are complementary to the nucleobases at the corresponding positions of the target nucleic acid by the total length of the oligomeric compound.

[0134] As used herein, "percent identity" refers to the number of nucleobases in a first nucleic acid that are of the same type (irrespective of chemical modification) as the nucleobases at the corresponding positions in a second nucleic acid, divided by the total number of nucleobases in the first nucleic acid.

[0135] As used herein, "modulation" refers to a change in the quantity or quality of a molecule, function, or activity as compared to the quantity or quality of the molecule, function, or activity before modulation. For example, modulation includes a change in gene expression, i.e., an increase (stimulation or induction) or a decrease (inhibition or reduction).

[0136] As used herein, "modification type" in reference to a nucleoside or "type" of nucleoside refers to a chemical modification of the nucleoside, including modified and unmodified nucleosides. Thus, unless otherwise specified, a "nucleoside having a first type of modification" can be an unmodified nucleoside.

[0137] As used herein, "different modifications" refers to chemical modifications or chemical substituents that are different from each other, including the absence of modification. Thus, for example, MOE nucleosides and unmodified naturally occurring RNA nucleosides are "differently modified", even though the naturally occurring nucleoside is unmodified. Similarly, DNA and RNA oligonucleotides are "differently modified", even though both are unmodified naturally occurring nucleosides. Nucleosides that are identical but contain different nucleobases do not have different modifications. For example, a nucleoside containing a 2'-OMe modified sugar moiety and an unmodified adenine nucleobase and a nucleoside containing a 2'-OMe modified sugar moiety and an unmodified thymine nucleobase do not have different modifications.

[0138] "Same type of modification" as used herein refers to modifications that are the same as each other, including the absence of modification. Thus, for example, two unmodified RNA nucleosides have the "same type of modification", even though the RNA nucleosides are unmodified. Such nucleosides with the same type of modification may contain different nucleobases.

[0139] "Region" or "portion" as used herein refers to a plurality of linked nucleosides having the functions or characteristics defined herein, particularly with reference to the claims and definitions provided herein. Generally, such a region or portion contains at least 10, at least 11, at least 12, or at least 13 linked nucleosides. For example, such a region may contain 13 to 20 linked nucleosides, such as 13 to 16 or 18 to 20 linked nucleosides. Generally, the first region defined herein consists essentially of 18 to 20 nucleosides, while the second region defined herein consists essentially of 13 to 16 linked nucleosides.

[0140] "Pharmaceutically acceptable carrier or diluent" as used herein refers to any substance suitable for administration to an animal. In certain embodiments, the pharmaceutically acceptable carrier or diluent is sterile saline. In certain embodiments, this sterile saline is pharmaceutical grade saline. "Substituent" and "substituent group" as used herein refer to an atom or group of atoms that replaces an atom or group of atoms of a specified parent compound. For example, a substituent of a modified nucleoside is any atom or group of atoms different from those found in a naturally occurring nucleoside (e.g., a modified 2'-substituent is any atom or group of atoms other than H or OH at the 2'-position of the nucleoside). A substituent group may be protected or unprotected. In certain embodiments, the compounds of the present disclosure have substituents at one or more positions of the parent compound. A substituent may further be substituted by other substituents and may be directly or through a linking group (such as oxygen or an alkyl or hydrocarbon group) attached to the parent compound.

[0141] Such substituents can be present as modifications on the sugar moiety, particularly substituents present at the 2'-position of the sugar moiety. Unless otherwise indicated, groups that can serve as substituents include, but are not limited to, one or more of halogen, hydroxy, alkyl, alkenyl, alkynyl, acyl, carboxyl, alkoxy, alkoxyalkylene, and amino substituents. Certain substituents described herein can represent modifications directly attached to the ring of the sugar moiety (e.g., halogen, such as fluorine, directly attached to the sugar ring), or modifications indirectly attached to the ring of the sugar moiety through an oxygen linking atom, which oxygen linking atom is itself directly attached to the sugar moiety (e.g., alkoxyalkylene, such as methoxyethylene, attached to an oxygen atom, generally providing an MOE substituent attached to the 2'-position of the sugar moiety as described herein).

[0142] As used herein, "alkyl" refers to a saturated straight-chain or branched-chain monovalent C1-6 hydrocarbon group, where methyl is the most preferred alkyl as a substituent at the 2'-position of the sugar moiety. The alkyl is typically attached to the oxygen linking atom at the 2'-position of the sugar, and thus generally provides an -O-alkyl substituent, such as an -OCH3 substituent, on the sugar moiety of the oligomeric compounds according to the present invention. This will be well understood by those skilled in the art.

[0143] As used herein, "alkylene" refers to a saturated straight-chain or branched-chain divalent hydrocarbon group of the general formula -C n H 2n -, where n is 1-6. Methylene or ethylene is the preferred alkylene.

[0144] As used herein, "alkenyl" refers to a straight-chain or branched-chain unsaturated monovalent C2-6 hydrocarbon group, where vinyl or propenyl is the most preferred alkenyl as a substituent at the 2'-position of the sugar moiety. It is well understood in the art that the unsaturation present in the alkenyl is the presence of at least one carbon-carbon double bond. The alkenyl is typically attached to the oxygen linking atom at the 2'-position of the sugar, and thus generally provides an -O-alkenyl substituent, such as an -OCH2CH=CH2 substituent, on the sugar moiety of the oligomeric compounds according to the present invention. This will be well understood by those skilled in the art.

[0145] As used herein, "alkynyl" refers to a straight-chain or branched-chain unsaturated C2-6 hydrocarbon group, where ethynyl is the most preferred alkynyl as a substituent at the 2'-position of the sugar moiety. As is well known in the art, the unsaturation present in the alkynyl is the presence of at least one carbon-carbon triple bond. The alkynyl is typically attached to the oxygen linking atom at the 2'-position of the sugar, and thus generally provides an -O-alkynyl substituent on the sugar moiety of the oligomeric compounds according to the present invention. This will be well understood by those skilled in the art.

[0146] As used herein, "carboxyl" is a group having the general formula -CO2H.

[0147] As used herein, "acyl" refers to a group formed by removing a hydroxyl group from a carboxyl group as defined herein, and has the general formula -C(O)-X, where X is usually a C1-6 alkyl group.

[0148] As used herein, "alkoxy" refers to a group formed between an alkyl group (such as a C1-6 alkyl group) and an oxygen atom, where the oxygen atom is used to connect the alkoxy group to the parent molecule (such as the 2'-position of a sugar moiety) or another group (such as an alkylene group as defined herein). Examples of alkoxy groups include, but are not limited to, methoxy, ethoxy, propoxy, isopropoxy, n-butoxy, sec-butoxy, and tert-butoxy. The alkoxy groups used herein may optionally include further substituents.

[0149] As used herein, alkoxyalkylene refers to an alkoxy group as defined herein that is connected to an alkylene group as defined herein, where the oxygen atom of the alkoxy group is connected to the alkylene group, and the alkylene group is connected to the parent molecule. The alkylene group is usually connected to an oxygen-bonded atom at the 2'-position of the sugar, and thus generally provides an -O-alkylene-alkoxy substituent on the sugar moiety of the oligomeric compound according to the present invention, such as an -OCH2CH2OCH3 substituent. Those skilled in the art will fully understand this, and it is generally referred to as an MOE substituent as defined herein and well-known in the art.

[0150] "Amino" as used herein includes primary, secondary, and tertiary amino groups.

[0151] "Halogen" and "halogens" as used herein refer to atoms selected from fluorine, chlorine, bromine, and iodine.

[0152] As used herein, the term "mxRNA" should be understood in particular as defined in WO 2020 / 044186 A2, which is incorporated herein by reference in its entirety. Specifically, mxRNA is a hairpin-shaped RNA molecule composed of an antisense portion (also called the guide strand) and a sense portion (also called the passenger strand). mxRNA includes a double-stranded region and a hairpin loop, where the length of the mxRNA is about 34 nucleotides. The double-stranded region includes a region where a portion of the antisense portion and substantially the entire sense portion (usually 14 or 15 nucleotides of each strand) are base-paired. The hairpin loop connects the two regions of the double strand, i.e., the antisense region and the sense region, through, for example, a phosphate or phosphorothioate linker (i.e., covalently), and the antisense portion usually has a length of about 18 to 20 nucleotides, thus forming an antisense double-stranded region and a loop. The antisense portion is part of the loop, which further connects the sense portion (forming the second strand of the loop) and the antisense portion.

[0153] The term "angiotensinogen" or the abbreviation "AGT", also known as SERPINA 8 or ANHU, is used in its ordinary meaning to denote a protein produced by the liver and is part of the renin-angiotensin-aldosterone system (RAAS). When released into the circulation, it is converted by renin into angiotensin I. Angiotensinogen is expressed and produced in the liver by the angiotensin gene or "AGT gene". As used herein, the term "muRNA" or "multi-RNA" includes nucleic acid constructs containing more than one (usually two) RNA sequences, namely a first and a second nucleic acid portion targeting different regions of the C5 mRNA; or a region of the C5 mRNA and a region of the mRNA of another target molecule. The RNA sequences targeted are also referred to as the "antisense" or "guide" strands, and the corresponding passenger strands (i.e., the third and fourth nucleic acid portions complementary to the first and second portions respectively) are also included in the nucleic acid construct. Specifically, such muRNAs are designed to be broken down and release the first and second nucleic acid portions after in vivo administration. A specific example of such a muRNA is shown below, where (1) is the first nucleic acid portion, (2) is the third nucleic acid portion complementary to (1), (3) is the second nucleic acid portion complementary to the fourth nucleic acid portion, and (5) is a labile linker, and (6) is a ligand, all of which will be explained below.

[0154]

[0155] It should also be understood that the oligomeric compounds described herein may have one or more non-hybridizing nucleosides (overhangs) and / or one or more internal non-hybridizing nucleosides (mismatches) at one or both ends of one or both strands, provided that there is sufficient complementarity to maintain hybridization under physiologically relevant conditions. Alternatively, the oligomeric compounds described herein may have blunt ends at at least one end.

[0156] The term "complement component C5" or simply "C5" as used herein denotes the corresponding, generally known protein, which is cleaved into C5a and C5b, where C5b forms part of the membrane attack complex in the later stages of complement activation. C5 is a protein encoded by the C5 gene in humans. Complement component C5 is the fifth component of the complement and plays an important role in the processes of inflammation and cell killing. The protein consists of α and β polypeptide chains linked by disulfide bonds. The activated peptide C5a is an anaphylatoxin with potent spasmogenic and chemotactic activities, obtained by cleavage of the α polypeptide by C5 convertase. The large molecular cleavage product of C5b can form a complex with the C6 complement component, which is the basis for the formation of the membrane attack complex including other complement components.

[0157] The term "comprising" as used herein means including the identified method steps or elements, but these steps or elements do not constitute an exclusive list, so there may be additional steps or elements.

[0158] In addition, to the extent that the term "comprising" is used in the detailed description or claims, that term is intended to be inclusive in a manner similar to the term "including" as "including" is construed as inclusive when used as a transitional word in a claim.

[0159] The present invention relates to the following aspects and embodiments

[0160] Small hairpin (shRNA) and mxRNA oligomeric compounds

[0161] According to a first aspect, the present invention relates to an oligomeric compound capable of inhibiting the expression of complement factor C5 (C5), wherein the compound comprises at least a first linked nucleoside region having at least a first nucleoside base sequence that is at least partially complementary to at least a portion of the RNA transcribed from the C5 gene, wherein the first nucleoside base sequence is selected from the sequences of Table 1a (SEQ ID NOs: 1 to 250) or a portion thereof, and wherein the portion preferably has a length of at least 18 nucleosides. Specifically, the 5'-terminal nucleoside of the first nucleoside base sequence may respectively comprise U instead of A; or U instead of G; or U instead of C.

[0162] In certain embodiments, the oligomeric compound may further comprise at least a second linked nucleoside region having at least a second nucleoside base sequence that is at least partially complementary to the first nucleoside base sequence and selected from the sequences of Table 1b (SEQ ID NOs: 251 to 500) or a portion thereof, and wherein the length of the portion is preferably at least 8, 9, 10 or 11 nucleosides, more preferably at least 10 nucleosides.

[0163] Specifically, the 3'-terminal nucleoside of the second nucleoside base sequence may respectively comprise A instead of U, G or C; and more specifically, the nucleobase A is the complementary nucleobase to the 5'-terminal nucleoside of the first nucleoside base sequence.

[0164] The first region of the linked nucleosides is also referred to as the antisense region or the guide region / strand, while the second region of the linked nucleosides is referred to as the sense region or the passenger region / strand. As disclosed in the preferred embodiments below, the two regions may be located on the same RNA strand, preferably in an adjacent manner. This results in a hairpin molecule, also referred to as mxRNA. On the other hand, the two regions may be located on different strands, which results in double-stranded RNA (dsRNA), wherein preferably each strand consists of the respective regions.

[0165] Without being bound by theory, it is hypothesized that the oligomeric compounds of the invention comprising the first linking nucleoside region and the second linking nucleoside region can be cleaved by the protein Argonaute 2 (Ago2) during RNA interference. The mxRNA is incorporated into the RNA-induced silencing complex (RISC). RISC assembly then binds and degrades the target mRNA. Specifically, this is achieved when the guide strand pairs with the complementary sequence in the C5 mRNA molecule and induces cleavage by Ago2, the catalytic component of RISC. Thus, since the expression of C5 is inhibited, it is believed that the effects associated with the overregulation of angiotensin II via the above pathway are also inhibited.

[0166] In certain embodiments, the first nucleobase sequence is selected from the following sequences or portions thereof: SEQ ID NO: 61, 30, 37, 87, 55, 66, 23, 83, 43, 47, 72, 27, 14, 28, 46, 82, 74, 75, 73, 53, 16, 36, 59, 42, and 56. Preferably, the first nucleobase sequence is selected from the following sequences or portions thereof: SEQ ID NO: 61, 30, 37, 83, 82, 74, 75, 73, 53, 16, 36, 59, 42, and 56, preferably 30, 37, and 83, more preferably 30 and 37.

[0167] In its preferred embodiments, the second nucleobase sequence is selected from the following sequences or portions thereof: SEQ ID NO: 311, 280,

[0168] 287, 333, 332, 324, 325, 323, 303, 266, 286, 309, 292, 306, and preferably 280, 287, and 333, more preferably 30 and 37.

[0169] Thus, the oligomeric compounds according to the invention can comprise SEQ ID NO 30 + 280 or 37 + 287.

[0170] Length and molecular characteristics of the oligomeric compounds according to the first aspect

[0171] The first linking nucleoside region can consist essentially of 18 to 35, preferably 18 to 20, more preferably 18 or 19, more preferably 19 linking nucleosides. Additionally, the second linking nucleoside region can consist essentially of 10 to 35, preferably 10 to 20, more preferably 10 to 16, more preferably 10 to 15, particularly 13, 14, or 15 linking nucleosides.

[0172] The oligomeric compound comprising the first and second linking nucleoside regions may comprise at least one complementary duplex region, which comprises at least a portion of the first linking nucleoside region directly or indirectly linked to at least a portion of the second linking nucleoside region, wherein preferably the length of the duplex region is from 10 to 19 base pairs, more preferably from 12 to 19 base pairs, and more preferably from 12 to 15 base pairs, especially 14 or 15 base pairs, wherein optionally there is one mismatch within the duplex region.

[0173] In certain embodiments, each of the first and second regions of the linking nucleoside has a 5' to 3' directionality, thereby defining its 5' and 3' regions, respectively.

[0174] In an oligomeric compound having the first linking nucleoside region and the second linking nucleoside region with 5' to 3' directionality, the 5' region of the first linking nucleoside region may be directly or indirectly linked to the 3' region of the second linking nucleoside region, for example by complementary base pairing, wherein preferably the 5'-terminal nucleoside of the first nucleoside region base pairs with the 3'-terminal nucleoside of the second nucleoside region.

[0175] In the above embodiments, the 3' region of the first linking nucleoside region may be directly or indirectly linked to the 5' region of the second linking nucleoside region, wherein preferably the first nucleoside region is directly covalently linked to the second nucleoside region by a phosphate, phosphorothioate or dithiophosphate, wherein more preferably the 3'-terminal nucleoside of the first linking nucleoside region is directly covalently linked to the 5'-terminal nucleoside of the second linking nucleoside region by a phosphate, phosphorothioate or dithiophosphate. Particularly preferably, the 3'-terminal nucleoside of the first region is directly linked to the 5'-terminal nucleoside of the second region by an internucleoside phosphorothioate bond.

[0176] This corresponds to forming a single oligonucleotide that comprises or consists of the two regions directly fused together. Due to the base pairing as defined in the previous embodiment, such an oligonucleotide will assume a hairpin structure. Optimized hairpins, especially in terms of size, are the subject of further embodiments below.

[0177] In certain embodiments, the oligomeric compound may consist of the first linking nucleoside region and the second linking nucleoside region. Each of the regions may constitute a separate strand, thereby yielding double-stranded RNA (dsRNA). A particularly preferred dsRNA of the invention is a dsRNA having a length of 19 nucleosides for the first strand and a length of 14 or 15, preferably 14, nucleosides for the second region. When used to define the length of a region or strand, the terms "nucleoside" and "nucleotide" (sometimes abbreviated as "nt") are used interchangeably.

[0178] Alternatively, as described above, the two regions can fuse together to form a hairpin structure.

[0179] In certain embodiments, there may be an intervening third region of linking nucleosides between the first region and the second region. In a preferred embodiment, the oligomeric compound comprises or consists of a single strand comprising the first, the third, and the second nucleoside regions, wherein at least a portion of the first nucleoside region is directly or indirectly linked to at least a portion of the second nucleoside region so as to form the at least partially complementary double-stranded region.

[0180] In other words, the oligomeric compound comprises a single strand comprising the first and second nucleoside regions, wherein at least a portion of the first nucleoside region is directly or indirectly linked to at least a portion of the second nucleoside region so as to form the at least partially complementary double-stranded region. As described above, the third region is optional.

[0181] In certain embodiments, the oligomeric compound may comprise or consist of a single strand that comprises or consists of the first and second linked nucleoside regions, wherein at least a portion of the first linked nucleoside region is directly or indirectly linked to at least a portion of the second linked nucleoside region so as to form the at least partially complementary double-stranded region.

[0182] In an oligomeric compound that may comprise or consist of a single strand, the first nucleoside region and the second nucleoside region are directly adjacent on the single strand.

[0183] In certain embodiments, the first nucleoside region may have a greater number of linking nucleosides compared to the second nucleoside region.

[0184] Optionally, the ratio of the total number of linking nucleosides in the first nucleoside region to the total number of linking nucleosides in the second nucleoside region ranges from about 19 / 15 to about 19 / 8 or from about 18 / 15 to about 18 / 8. In a particularly preferred embodiment, the ratio is 19 / 15, 19 / 14, 19 / 13, 18 / 15, 18 / 14, or 18 / 13, most preferably 19 / 14 or 19 / 15.

[0185] Optionally or alternatively, the percentage of the total number of linking nucleosides in the first nucleoside region relative to the total number of nucleosides in the oligomeric compound may range from about 55% to about 60%. In a particularly preferred embodiment, the percentage range may be from about 57% to about 59.5%, and most preferably, the percentage is about 57.6% or about 59.4%.

[0186] Without being bound by theory, it is assumed that the above ratios and / or percentages provide an appropriate ratio / percentage of the number of nucleotides in the antisense (guide) strand to the number of nucleotides in the sense (passenger) strand such that it can be processed by the RISC complex as described above without being significantly degraded beforehand, and thus, C5 knockdown can be effectively carried out.

[0187] In an oligomeric compound in which the number of linking nucleotides in the first region is greater than the number of linking nucleotides in the second region, an additional number of linking nucleotides in the first nucleoside region form a hairpin loop that links the first and second linking nucleoside regions, wherein preferably, a first nucleoside base sequence that is part of the first nucleoside base sequence of the complementary RNA transcribed from the C5 gene forms the hairpin loop, and wherein the loop comprises 2 to 5, preferably 4 or 5 nucleosides.

[0188] Such compounds are also referred to herein as hairpins or mxRNAs. Since the second region is shorter than the first region, the size of the compound is optimized (or miniaturized) compared to a conventional siRNA having two regions of comparable length.

[0189] Preferably, the loop has 4 or 5 linked nucleosides. Particularly preferred is that the first region has a length of 19 nucleosides, the second region has a length of 14 nucleosides, and the hairpin loop has a length of 5 nucleosides, wherein the 5 nucleosides in the hairpin are the 5 3'-terminal nucleosides of the first region. This molecular structure of the hairpin or mxRNA of the present invention is also referred to herein as "14-5-14". In certain embodiments, the oligomeric single strand can be selected from Table 2 as disclosed previously herein, particularly SEQ ID NO: 561, 530, 537, 587, 555, 566, 523, 583, 543, 547, 572, 527, 514, 528, 546, 582, 574, 575, 573, 553, 516, 536, 559, 542, and 556, preferably 583, 530, and 537, more preferably 530 and 537, wherein preferably the 5'-terminal nucleoside of the first linking nucleoside region comprises U as the nucleoside base, and the 5'-terminal nucleoside of the second linking nucleoside region comprises as the nucleoside base.

[0190] In a specific embodiment, the single strand is selected from Table 3c, particularly selected from construct ID NO: 1011, 980, 987, 1037, 1005, 1016, 973, 1033, 993, 997, 1022, 977, 964, 978, 996, 1032, 1024, 1025, 1023

[0191] 1003, 966, 986, 1009, 992, 1006, preferably 980, 987 and 1033, more preferably 980 and 987, wherein preferably the 5'-terminal nucleoside of the first linking nucleoside region comprises U as the nucleobase, and the 5'-terminal nucleoside of the second linking nucleoside region comprises A as the nucleobase.

[0192] In certain embodiments, a hairpin loop as previously described herein may be present in the 3'-region of the first linking nucleoside region, wherein optionally one, two or more 3'-terminal nucleosides of the first nucleobase sequence, within the scope permitted by the nucleobases of the one, two or more 3'-terminal nucleosides, fold back and form or contribute to the second linking nucleoside region.

[0193] This is a structural design, also known as "spillover". Spillover is only possible if there is self-complementarity between the nucleobases at the 3'-end of the guide sequence region contained in the double strand and the nucleobases of the 3'-terminal nucleosides of the same guide sequence. For example, this can be achieved as a 13-5-13 design, thus allowing further miniaturization. The first "13" refers to the guide sequence region involved in the double strand, 5 is the length of the loop formed by the guide sequence, and the second 13 refers to the second region of the double strand, formed by one nucleobase of the guide sequence and 12 nucleobases of the passenger region in the 5' to 3' direction. Thus, the length of the guide sequence remains 19 nucleosides, but the passenger sequence is shortened to 12 nucleosides.

[0194] In certain embodiments, in the case of the third nucleoside region as previously described herein, the third nucleoside region and optionally the 3'-terminal portion of the first nucleoside region (preferably consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 linked nucleosides) and / or the 5'-terminal portion of the second nucleoside region (preferably consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 linked nucleosides) may form a hairpin loop.

[0195] In certain embodiments, the hairpin loop comprises 1 to 8, 2 to 7, 3 to 6, preferably 4 or 5 linked nucleosides. The oligomeric compound according to the first aspect disclosed herein may be blunt-ended.

[0196] In the oligomeric compound according to the first aspect disclosed herein, the first or second nucleoside region may have an overhang.

[0197] In the oligomeric compound according to the first aspect disclosed herein, the first region may be selected from the sequences of Table 3a, or portions thereof, particularly selected from construct ID NO: 327, 352, 356, 362, 375 and 393.

[0198] In the oligomeric compound according to the first aspect disclosed herein, the second region may be selected from the sequences of Table 3b, or portions thereof, particularly portions of length 14 nucleosides, especially from constructs ID NO: 427, 452, 456, 462, 475, and 493. The oligomeric compound may have a total length of from about 25 to about 35 nucleosides, particularly about 33 or about 34 nucleosides.

[0199] In certain embodiments, the terminal nucleoside at the 5' position of the first region has a nucleobase selected from the group consisting of A, U, G, and C, preferably U, and wherein optionally, the terminal nucleoside at the 3' position of the second region has a base complementary to the base at the 5' position of the first region, preferably A.

[0200] Ligand

[0201] The oligomeric compound may comprise one or more ligands.

[0202] The one or more ligands, particularly two or more or three ligands, may be conjugated to the second linked nucleoside region and / or the first linked nucleoside region.

[0203] The one or more ligands may bind in the 3' region, preferably to the 3' terminal nucleoside of the second linked nucleoside region and / or the first linked nucleoside region, and / or to the 5' terminal nucleoside of the second linked nucleoside region. Specifically, the ligand may bind to the 3' terminal nucleoside.

[0204] The one or more ligands are any cell targeting moieties, such as lipids, carbohydrates, aptamers, vitamins, and / or peptides that bind to specific targets on the cell membrane or cell surface.

[0205] The one or more ligands may include one or more, particularly three, carbohydrates.

[0206] The one or more, particularly three, carbohydrates may be monosaccharides, disaccharides, trisaccharides, tetrasaccharides, oligosaccharides, or polysaccharides.

[0207] The one or more carbohydrates may include or consist of one or more (particularly three) hexose moieties.

[0208] The one or more, particularly three, hexose moieties are one or more galactose moieties, one or more lactose moieties, one or more, particularly three, N-acetylgalactosamine moieties, and / or one or more mannose moieties.

[0209] The one or more carbohydrates may include one or more, particularly three, N-acetylgalactosamine moieties.

[0210] Alternatively, the one or more carbohydrates may comprise two or more N-acetylgalactosamine moieties, preferably three.

[0211] The one or more ligands are attached to the oligomeric compound in a linear configuration or a branched configuration, preferably to the second region of the oligomeric compound that links the nucleosides.

[0212] Particularly preferred ligands are as follows, also known as "toothbrushes":

[0213]

[0214] Without being bound by a particular theory, it is speculated that due to such ligands, the target tissue, i.e., the liver that produces C5, can be selectively targeted, and thus the oligomeric compound can more effectively exhibit inhibition of the C5 gene.

[0215] The one or more, particularly three, ligands may be attached to the oligomeric compound in a bi-antennary or tri-antennary structure.

[0216] The one or more ligands as described above are preferably attached to the 3'-terminal nucleoside of the second linking nucleoside region.

[0217] Internucleoside linkage

[0218] The oligomeric compound according to the first aspect disclosed herein may comprise internucleoside bonding, and at least one of the internucleoside bondings is a modified internucleoside bonding.

[0219] The modified internucleoside bonding may be a phosphorothioate or dithiophosphonate internucleoside bonding.

[0220] The oligomeric compound according to the first aspect disclosed herein may comprise 1 to 16 phosphorothioate or dithiophosphonate internucleoside bonds. Preferred modified internucleoside bonds are the subject of the following preferred embodiments. Certain modified internucleoside bonds are known in the art and are described, for example, in Signal Transduction and Targeted Therapy (2020) 5:101 by Hu et al.

[0221] The oligomeric compound may comprise 7, 8, 9, or 10 phosphorothioate or dithiophosphonate internucleoside bonds, and the one or more phosphorothioate or dithiophosphonate internucleoside bonds may be present in the 5'-region of the first linking nucleoside region, wherein preferably, the oligomeric compound comprises three phosphorothioate internucleoside bonds at three adjacent nucleosides in the 5'-region.

[0222] In addition, the oligomeric compound may comprise phosphorothioate or phosphorodithioate internucleoside linkages between at least two, preferably at least three, preferably at least four, preferably at least five adjacent nucleosides in the hairpin loop, depending on the number of nucleosides present in the hairpin loop. Specifically, the oligomeric compound may comprise phosphorothioate or phosphorodithioate internucleoside linkages between each adjacent nucleoside present in the hairpin loop.

[0223] Modification

[0224] In the oligomeric compound according to the first aspect of the present invention, at least one nucleoside comprises a modified sugar.

[0225] The modified sugar may be selected from 2'-modified sugars, conformationally restricted nucleoside (CRN) sugars such as locked nucleic acid (LNA) sugars, (S)-restricted ethyl bicyclic nucleic acid, restricted ethyl (cEt) sugars, tricyclo-DNA, morpholinos, unlocked nucleic acid (UNA) sugars, glycol nucleic acid (GNA), D-hexitol nucleic acid (HNA), cyclohexene nucleic acid (CeNA).

[0226] Preferred modified sugars are the subject of the following preferred embodiments. Certain modified sugars are known in the art and are described, for example, in Hu et al., Signal Transduction and Targeted Therapy (2020) 5:101.

[0227] The 2'-modified sugar may be selected from 2'-O-alkyl modified sugars, 2'-O-methyl modified sugars, 2'-O-methoxyethyl modified sugars, 2'-O-allyl modified sugars, 2'-C-allyl modified sugars, 2'-deoxy modified sugars such as 2'-deoxyribose, 2'-F modified sugars, 2'-arabino-fluoro modified sugars, 2'-O-benzyl modified sugars, 2'-O-methyl-4-pyridyl modified sugars, wherein at least one modified sugar may be a 2'-O-methyl modified sugar.

[0228] At least one modified sugar may be a 2'-F modified sugar, and preferably, at most 16 or 17 sugars are 2'-F modified sugars. Preferably, the sugar is ribose.

[0229] In the oligomeric compound according to the first aspect of the present invention, the sugars of the nucleosides at any position from 2 to 14 downstream of the first nucleoside in the 5' region of the first linked nucleoside region do not contain a 2'-O-methyl modification.

[0230] In certain embodiments, the 3' terminal position of the second region of the linked nucleoside does not contain a 2'-O-methyl modification.

[0231] In certain embodiments, the sugar of the nucleoside at either position 2 or 14 downstream of the first nucleoside in the 5' region of the first linked nucleoside region contains a 2'-F modification.

[0232] In certain embodiments, the sugar of a nucleoside in the second linked nucleoside region corresponding to any nucleoside in the first linked nucleoside region is located at any position among positions 11 to 13 downstream of the first nucleoside in the 5' region of the first linked nucleoside region and contains a 2'-F modification.

[0233] In certain embodiments, the 3'-terminal nucleoside of the second linked nucleoside region contains a 2'-F modification.

[0234] In certain embodiments, one or more of the odd-numbered nucleosides starting from the 5' region of the first linked nucleoside region can be modified, and / or one or more of the even-numbered nucleosides starting from the 5' region of the first linked nucleoside region can be modified, wherein the modification of the even-numbered nucleosides is typically a second modification different from the modification of the odd-numbered nucleosides.

[0235] In certain embodiments, one or more odd nucleosides starting from the 3' region of the second linked nucleoside region can be modified by a modification different from the modification of the odd nucleosides in the first linked nucleoside region.

[0236] In certain embodiments, one or more even nucleosides starting from the 3' region of the second linked nucleoside region are modified by a modification different from the modification of the even nucleosides in the first linked nucleoside region.

[0237] In certain embodiments, at least one or more modified even-numbered nucleosides in the first linked nucleoside region are adjacent to at least one or more differently modified odd-numbered nucleosides in the first nucleoside region.

[0238] In certain embodiments, at least one or more modified even-numbered nucleosides in the second nucleoside region are adjacent to at least one or more differently modified odd-numbered nucleosides in the second linked nucleoside region.

[0239] In certain embodiments, the sugar of one or more of the odd nucleosides starting from the 5' region of the first nucleoside region can be a 2'-O-methyl-modified sugar.

[0240] In certain embodiments, one or more of the even nucleosides starting from the 3' region of the first region of the linked nucleoside can be a 2'-F-modified sugar.

[0241] In certain embodiments, the sugar of one or more of the odd nucleosides starting from the 5' region of the second region of the linked nucleoside can be a 2'-O-methyl-modified sugar.

[0242] In certain embodiments, one or more of the even nucleosides starting from the 5' region of the second linked nucleoside region can be a 2'-F-modified sugar.

[0243] In certain embodiments, the sugars of multiple adjacent nucleosides in the first nucleoside region can be modified by common or different modifications.

[0244] In certain embodiments, the sugars of multiple adjacent nucleosides in the second nucleoside region can be modified by common or different modifications.

[0245] In certain embodiments, the sugars of multiple adjacent nucleosides in the hairpin loop can be modified by common modification or different modifications, and the common modification can be a 2'-F modified sugar.

[0246] Alternatively, the common modification can be a 2'-O-methyl modified sugar.

[0247] The multiple adjacent 2'-O-methyl modified sugars can be present in at least eight adjacent nucleosides in the first and / or second nucleoside regions. The multiple adjacent 2'-O-methyl modified sugars can be present in three or four adjacent nucleosides in the hairpin loop.

[0248] In certain embodiments, the hairpin loop (as disclosed hereinbefore) can comprise at least one nucleoside having a modified sugar.

[0249] In certain embodiments, the at least one nucleoside is adjacent to a nucleoside having a sugar with a different modification, and preferably all adjacent nucleosides in the hairpin loop have sugars with different modifications.

[0250] In certain embodiments, the modified sugar is a 2'-O-methyl modified sugar, and the differently modified sugar is a 2'-F modified sugar.

[0251] In certain embodiments, one or more nucleosides in the first linking nucleoside region and / or the second linking nucleoside region can be inverted nucleosides and are linked to adjacent nucleosides via the 3'-carbon of their sugars and the 3'-carbon of the sugars of adjacent nucleosides, and / or one or more nucleosides in the first linking nucleoside region and / or the second linking nucleoside region are inverted nucleosides and are linked to adjacent nucleosides via the 5'-carbon of their sugars and the 5'-carbon of the sugars of adjacent nucleosides.

[0252] muRNA nucleic acid constructs

[0253] According to a second aspect, the present invention relates to a nucleic acid construct comprising at least:

[0254] (a) A first nucleic acid portion that is at least partially complementary to at least a first portion of an RNA transcribed from the C5 gene;

[0255] (b) A second nucleic acid portion that is at least partially complementary to at least a second portion of an RNA transcribed from the C5 gene, the second portion being different from the first portion;

[0256] (c) A third nucleic acid portion that is at least partially complementary to the first nucleic acid portion of (a) such that a first nucleic acid duplex region is formed therewith;

[0257] (d) A fourth nucleic acid portion that is at least partially complementary to the second nucleic acid portion of (b) such that a second nucleic acid duplex region is formed therewith.

[0258] The construct can be designed such that after in vivo administration, the construct disassembles to produce at least first and second discrete nucleic acid targeting molecules that respectively target the RNA portions transcribed from the target genes of (a) and (b);

[0259] wherein (i) the first nucleic acid targeting molecule is capable of regulating the expression of the target gene of (a) and comprises or is derived from at least the first nucleic acid portion of (a), and (ii) the second nucleic acid targeting molecule is capable of regulating the expression of the target gene of (b) and comprises or is derived from the second nucleic acid portion of (b).

[0260] The construct can be designed to disassemble such that the first and second discrete nucleic acid targeting molecules are respectively processed by independent RNAi-induced silencing complexes.

[0261] Sequence characteristics, labile functions, and structural characteristics of the RNA molecule

[0262] The construct according to the second aspect and its foregoing embodiments can comprise at least one labile function such that after in vivo administration, the construct is cleaved to produce the at least first and second discrete nucleic acid targeting molecules.

[0263] The labile function can comprise one or more unmodified nucleotides. Specifically, the one or more unmodified nucleotides of the labile function represent one or more cleavage positions in the construct, such that after in vivo administration, the construct is cleaved at the one or more cleavage positions to produce the at least first and second discrete nucleic acid targeting molecules. In particular, the cleavage positions can be respectively located in the construct such that after cleavage, the first discrete nucleic acid targeting molecule comprises or is derived from the first nucleic acid duplex region, and the second discrete nucleic acid targeting molecule comprises or is derived from the second nucleic acid duplex region. Preferably, the first discrete nucleic acid targeting molecule comprises or consists of the first nucleic acid portion of (a) and the third nucleic acid portion of (c), and / or the second discrete nucleic acid targeting molecule comprises or consists of the second nucleic acid portion of (b) and the fourth nucleic acid portion of (d).

[0264] In certain embodiments

[0265] (a) The first nucleic acid portion has a nucleobase sequence selected from SEQ ID NOs: 1 to 250 in Table 1a;

[0266] (b) The second nucleic acid portion has a nucleobase sequence selected from Table 1a (SEQ ID NOs: 1 to 250);

[0267] (c) The third nucleic acid portion has a nucleobase sequence selected from SEQ ID NOs: 251 to 500 in Table 1b; and / or

[0268] (d) The fourth nucleic acid portion has a nucleobase sequence selected from Table 1b (SEQ ID NOs: 251 to 500).

[0269] Wherein the third and fourth nucleobase sequences may be one, two, or three nucleobases shorter within the range of their 14-nucleobase length, and preferably the 5'-terminal nucleobase is absent.

[0270] In certain such embodiments, the first nucleic acid portion of (a) may be directly or indirectly linked to the fourth nucleic acid portion of (d) as a primary structure.

[0271] In certain embodiments, the first and fourth nucleic acid portions have nucleobase sequences of SEQ ID NO: 30 and 287, 30 and 333, 37 and 280, 37 and 333, 83 and 280, 83 and 287, respectively, and preferably, wherein the

[0272] sequences of SEQ ID NO: 280, 287, and 333 may be one, two, three, or four nucleobases shorter, and preferably the 5'-terminal nucleobase is absent.

[0273] In certain embodiments, the second nucleic acid portion of (b) may be directly or indirectly linked to the third nucleic acid portion of (c) as a primary structure.

[0274] In certain embodiments, the second and third nucleic acid portions have nucleobase sequences of SEQ ID NO: 30 and 287, 30 and 333, 37 and 280, 37 and 333, 83 and 280, 83 and 287, respectively, and preferably, wherein the sequences of SEQ ID NO: 280, 287, and 333 may be one, two, three, or four nucleobases shorter, and preferably the 5'-terminal nucleobase is absent.

[0275] In certain embodiments, the construct may further comprise 1 to 8 additional nucleic acid moieties, each of which is at least partially complementary to a respective additional 1 to 8 RNA moieties transcribed from one or more target genes, which target genes may be the same as or different from each other, and / or the same as or different from the target genes defined in (a) and / or (b), and wherein each of the 1 to 8 additional nucleic acid moieties forms an additional duplex region with a respective passenger nucleic acid moiety that is at least partially complementary thereto. Specifically, the second nucleic acid moiety of (b) and the 1 to 8 additional nucleic acid moieties may be directly or indirectly linked to a selected passenger nucleic acid moiety as their respective primary structures.

[0276] In certain embodiments, the direct or indirect linkage may represent (i) an internucleotide bond, (ii) an internucleotide gap, or (iii) a nucleic acid linker moiety of 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 nucleotides, which nucleic acid linker is preferably single-stranded. Preferably, the linkage may be direct, resulting in a continuous strand.

[0277] In certain embodiments, there may be some complementarity between the first nucleic acid moiety of (a) and the second nucleic acid moiety of (b), or between the third nucleic acid moiety of (c) and the fourth nucleic acid moiety of (d). Preferably, the complementarity (i) may be 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, preferably 2, 3, 4, or 5 base pairs; and / or

[0278] (ii) may be located between the first nucleic acid moiety of (a) and the second nucleic acid moiety of (b).

[0279] In certain embodiments, the internucleotide bond may involve at least one of the one or more unmodified nucleotides, wherein preferably cleavage may occur at the 3' position of the unmodified nucleotide(s).

[0280] In certain embodiments, the length of the first nucleic acid moiety of (a) and / or the second nucleic acid moiety of (b) and / or the third nucleic acid moiety of (c) and / or the fourth nucleic acid moiety of (d) is 7 to 25 nucleotides, respectively. Optionally, the length of the first nucleic acid moiety of (a) and / or the second nucleic acid moiety of (b) may be 18 to 21 nucleotides, more preferably 18 to 20 nucleotides, more preferably 19 nucleotides. In a preferred embodiment, the length of the first nucleic acid moiety of (a) and the second nucleic acid moiety of (b) is 19 nucleotides. Further preferably, the length of the third nucleic acid moiety of (c) and / or the fourth nucleic acid moiety of (d) is 11 to 20 nucleotides, more preferably 13 to 16 nucleotides, more preferably 14 or 15 nucleotides, and most preferably 14 nucleotides.

[0281] In certain embodiments, the first nucleic acid portion of (a) and the second nucleic acid portion of (b) can have a length of 19 nucleotides, and the third nucleic acid portion of (c) and the fourth nucleic acid portion of (b) can have a length of 14 nucleotides.

[0282] In certain embodiments, the unmodified nucleotide is located at any position from position 18 to 25, more preferably at any position from position 18 to 21, and / or at the 3'-terminal position of the first nucleic acid portion of (a) and / or the third nucleic acid portion of (c).

[0283] In certain embodiments, the unmodified nucleotide is located at position 19.

[0284] In certain embodiments, the first nucleic acid portion of (a) and the second nucleic acid portion of (b) can have a length of 19 nucleotides, while the third nucleic acid portion of (c) and the fourth nucleic acid portion of (b) can have a length of 14 nucleotides, and the unmodified nucleotide is located at the 19th position of the first nucleic acid portion of (a) and the second nucleic acid portion of (b).

[0285] In certain embodiments, the nucleic acid linker portion can have a length of 1 to 8 nucleotides, preferably a length of 2 to 7 or 3 to 6 nucleotides, more preferably a length of about 4 or 5 nucleotides and most preferably a length of 4 nucleotides.

[0286] In certain embodiments, one or more of all the duplex regions can independently have a length of 10 to 19 base pairs, more preferably 13 to 19 base pairs, and still more preferably 13, 14 or 15 base pairs, most preferably 14 base pairs, where optionally, there is one mismatch within the duplex region.

[0287] In certain embodiments, the nucleic acid construct can be blunt-ended.

[0288] In certain embodiments,

[0289] the first nucleic acid portion of (a); and / or

[0290] the second nucleic acid portion of (b); and / or

[0291] the third nucleic acid portion of (c); and / or

[0292] the fourth nucleic acid portion of (d); and / or

[0293] to the extent present, 1 to 8 additional nucleic acid portions as previously defined herein; and / or

[0294] To the extent of its existence, the passenger nucleic acid portion is as defined previously herein;

[0295] There may be an overhang.

[0296] In certain embodiments, the target RNA can be mRNA or another RNA molecule.

[0297] Ligand

[0298] The nucleic acid construct according to the second aspect and the foregoing embodiments may further comprise one or more ligands.

[0299] In certain embodiments, the first nucleic acid portion of (a), and / or the second nucleic acid portion of (b), and / or the third nucleic acid portion of (c), and / or the fourth nucleic acid portion of (d), and / or, to the extent of its existence, the 1 to 8 additional nucleic acid portions defined previously herein, and / or the passenger nucleic acid portion defined previously herein, may each have 5' to 3' directionality, thereby defining its 5' and 3' regions.

[0300] In certain embodiments, one or more ligands are conjugated to (i) the third nucleic acid portion of (c), and / or (ii) the fourth nucleic acid portion of (d), and / or, to the extent of its existence, the 3' region, preferably the 3' end, of the (iii) passenger nucleic acid portion as defined previously herein.

[0301] In certain embodiments, one or more ligands may be conjugated to one or more regions intermediate the 5' and 3' regions of any of the nucleic acid portions, preferably the third nucleic acid portion of (c), and / or the fourth nucleic acid portion of (d), and / or the passenger nucleic acid portion as defined previously herein.

[0302] In certain embodiments, one or more ligands may bind to the 5' region, preferably the 5' end, of any of the nucleic acid portions.

[0303] In certain embodiments, the one or more ligands can be any cell targeting moiety, such as a lipid, carbohydrate, aptamer, vitamin, and / or a peptide that binds to a specific target on the cell membrane or cell surface. In a preferred embodiment, the one or more carbohydrates can be a monosaccharide, disaccharide, trisaccharide, tetrasaccharide, oligosaccharide, or polysaccharide. In a more preferred embodiment, the one or more carbohydrates can include one or more hexose moieties. In particular, the one or more hexose moieties can be one or more galactose moieties, one or more lactose moieties, one or more N-acetylgalactosamine moieties, and / or one or more mannose moieties. The hexose moiety can include two or three N-acetylgalactosamine moieties. In particular, the hexose moiety can include three N-acetylgalactosamine moieties.

[0304] In certain embodiments, the one or more ligands may be linked in a linear configuration or a branched configuration. Preferably, wherein the one or more ligands may be linked in a bi-antennary configuration or a tri-antennary configuration, or in a configuration based on individual ligands at different positions. Preferably, the ligand may have the following structure:

[0305]

[0306] Internucleoside linkage

[0307] The nucleotide construct according to the second aspect of the present invention or its foregoing embodiments may comprise one or more phosphorothioate or phosphorodithioate internucleotide linkages.

[0308] In certain embodiments, the nucleic acid construct may comprise 1 to 15 phosphorothioate or phosphorodithioate internucleotide linkages.

[0309] In certain embodiments, the nucleic acid construct may comprise one or more phosphorothioate or phosphorodithioate internucleotide linkages at one or more 5' and / or 3' regions of the first nucleic acid portion of (a), and / or the second nucleic acid portion of (b), and / or the third nucleic acid portion of (c), and / or the fourth nucleic acid portion of (d), and / or the 1 to 8 additional nucleic acid portions as defined hereinbefore, and / or the passenger nucleic acid portion as defined hereinbefore.

[0310] In certain embodiments, the nucleic acid construct may comprise a phosphorothioate or phosphorodithioate internucleotide linkage between at least two adjacent nucleotides of the nucleic acid linker portion as defined hereinbefore.

[0311] In certain embodiments, the nucleic acid construct may comprise a phosphorothioate or phosphorodithioate internucleotide linkage between each adjacent nucleotide present in the nucleic acid linker portion.

[0312] In certain embodiments, the nucleic acid construct may comprise a phosphorothioate or phosphorodithioate internucleotide linkage connecting:

[0313] The first nucleic acid portion of (a) to the nucleic acid linker portion as previously defined herein; and / or

[0314] The second nucleic acid portion of (b) to the nucleic acid linker portion as previously defined herein; and / or

[0315] The third nucleic acid portion of (c) to the nucleic acid linker portion as previously defined herein and / or

[0316] The fourth nucleic acid portion of (d) to the nucleic acid linker portion as previously defined herein; and / or

[0317] Linking 1 to 8 additional nucleic acid moieties as defined previously herein to a nucleic acid linker moiety as further defined previously herein; and / or

[0318] A passenger nucleic acid moiety as defined previously herein and a nucleic acid linker moiety as further defined previously herein.

[0319] Modify

[0320] In the nucleic acid constructs according to the second aspect of the invention and its foregoing embodiments, at least one nucleotide of at least one of the following can be modified:

[0321] (a) the first nucleic acid moiety; and / or

[0322] (b) the second nucleic acid moiety; and / or

[0323] (c) the third nucleic acid moiety; and / or

[0324] (d) the fourth nucleic acid moiety; and / or

[0325] To the extent present, 1 to 8 additional nucleic acid moieties as defined previously herein; and / or

[0326] To the extent present, the passenger nucleic acid moiety as defined previously herein; and / or

[0327] If present, the nucleic acid linker moiety as further defined previously herein.

[0328] In a preferred embodiment, one or more odd-numbered nucleotides starting from the 5'-region of one of the following can be modified, and / or one or more even-numbered nucleotides starting from the 5'-region of one of the following can be modified, wherein the modification of the usually even-numbered nucleotides is a second modification different from the modification of the odd-numbered nucleotides:

[0329] (a) the first nucleic acid moiety; and / or

[0330] (b) the second nucleic acid moiety; and / or

[0331] (c) the third nucleic acid moiety; and / or

[0332] (d) the fourth nucleic acid moiety; and / or

[0333] To the extent present, 1 to 8 additional nucleic acid moieties as defined previously herein; and / or

[0334] To the extent present, the passenger nucleic acid moiety as defined previously herein.

[0335] In certain embodiments, one or more of the odd-numbered nucleotides starting from the 3' region of the third nucleic acid portion of (c) may be modified by a modification different from the modification of the odd-numbered nucleotides starting from the 5' region of the first nucleic acid portion of (a); and / or

[0336] one or more of the odd-numbered nucleotides starting from the 3' region of the fourth nucleic acid portion of (d) may be modified by a modification different from the modification of the odd-numbered nucleotides starting from the 5' region of the second nucleic acid portion of (b); and / or

[0337] One or more odd-numbered nucleotides starting from the 3' region of the passenger nucleic acid portion as previously defined herein, to the extent present, may be modified by a modification different from the modification of the odd-numbered nucleotides starting from the 5' region of 1 to 8 additional nucleic acid portions as previously defined herein; and / or

[0338] wherein one or more nucleotides of the nucleic acid linker portion, as further defined hereinabove, to the extent present, may be modified by a modification that (i) is different from the modification of the adjacent nucleotides in the 3' region of the first nucleic acid portion of (a); and / or (ii) is different from the modification of the adjacent nucleotides in the 3' region of the second nucleic acid portion of (b); and / or is different from the modification of the adjacent nucleotides in the 3' region of 1 to 8 additional nucleic acid portions, to the extent present, as previously defined herein.

[0339] In certain embodiments, one or more of the even-numbered nucleotides starting from the 3' region of: (i) the third nucleic acid portion of (c), and / or (ii) the fourth nucleic acid portion of (d), and / or (iii) the passenger nucleic acid portion as previously defined herein, to the extent present, may be modified by a modification different from the modification of the odd-numbered nucleotides starting from the 3' region of these respective portions.

[0340] In certain embodiments, (i) the first nucleic acid portion of (a), and / or (ii) the second nucleic acid portion of (b), and / or (iii) at least one or more modified even-numbered nucleotides of, to the extent present, 1 to 8 additional nucleic acid portions as previously defined herein, may be adjacent to at least one or more odd-numbered nucleotides modified differently from these respective portions.

[0341] In certain embodiments, (i) the third nucleic acid portion of (c), and / or (ii) the fourth nucleic acid portion of (d), and / or (iii) at least one or more of the modified even-numbered nucleotides of, to the extent present, the passenger nucleic acid portion as previously defined herein, may be adjacent to at least one or more odd-numbered nucleotides modified differently from these respective portions.

[0342] In certain embodiments, the first nucleic acid portion of (i)(a), and / or the second nucleic acid portion of (ii)(b), and / or the plurality of adjacent nucleotides of (iii), to the extent present, 1 to 8 additional nucleic acid portions as previously defined herein, can be modified by co-modification.

[0343] In certain embodiments, the third nucleic acid portion of (i)(c), and / or the fourth nucleic acid portion of (ii)(d), and / or the plurality of adjacent nucleotides of (iii), to the extent present, the passenger nucleic acid portion as previously defined herein, can be modified by co-modification.

[0344] In certain embodiments, the plurality of adjacent co-modified nucleotides can be 2 to 4 adjacent nucleotides, preferably 3 or 4 adjacent nucleotides.

[0345] In certain embodiments, the plurality of adjacent co-modified nucleotides can be located in the third nucleic acid portion of (i)(c), and / or the fourth nucleic acid portion of (ii)(d), and / or the 5' region of (iii), with respect to the passenger nucleic acid portion that was previously present herein.

[0346] In certain embodiments, the plurality of adjacent co-modified nucleotides can be located in a nucleic acid linker portion, as further defined hereinabove.

[0347] In certain embodiments, one or more modified nucleotides of the first nucleic acid portion of (a) may not have the co-modification present in the corresponding nucleotide of the third nucleic acid portion of (c) in the first duplex region; and / or one or more modified nucleotides of the second nucleic acid portion of (b) may not have the co-modification present in the corresponding nucleotide of the fourth nucleic acid portion of (d) in the second duplex region; and / or one or more modified nucleotides of 1 to 8 additional nucleic acid portions, to the extent present as previously defined herein, may not have the co-modification present in the corresponding nucleotide of the corresponding passenger nucleic acid portion in the respective duplex region.

[0348] In certain embodiments, one or more modified nucleotides of the first nucleic acid portion of (a) can be shifted by at least one nucleotide relative to the co-modified nucleotides of the third nucleic acid portion of (c); and / or one or more modified nucleotides of the second nucleic acid portion of (b) can be shifted by at least one nucleotide relative to the co-modified nucleotides of the fourth nucleic acid portion of (d); and / or one or more modified nucleotides of 1 to 8 additional nucleic acid portions (to the extent present as defined hereinbefore) can be shifted by at least one nucleotide relative to the co-modified nucleotides of the passenger nucleic acid portion (to the extent present as defined hereinbefore).

[0349] In certain embodiments, the modification and / or the modification may each individually be a sugar, phosphate or base modification.

[0350] In certain embodiments, the modification can be selected from nucleotides with 2' modified sugars; conformationally restricted nucleotide (CRN) sugars, such as locked nucleic acids (LNA), (S)-restricted ethyl bicyclic nucleic acids and restricted ethyl (cEt), tricyclic-DNA; morpholino, unlocked nucleic acids (UNA), glycol nucleic acids (GNA), D-hexitol nucleic acids (HNA) and cyclohexene nucleic acids (CeNA). In preferred embodiments, the 2' modified sugar can be selected from 2'-O-alkyl modified sugars, 2'-O-methyl modified sugars, 2'-O-methoxyethyl modified sugars, 2'-O-allyl modified sugars, 2'-C-allyl modified sugars, 2'-deoxy modified sugars such as 2'-deoxyribose, 2'-F modified sugars, 2'-arabinofluoride modified sugars, 2'-O-benzyl modified sugars, 2'-amino modified sugars and 2'-O-methyl-4-pyridine modified sugars.

[0351] In certain embodiments, the base modification may be any of an abasic nucleotide and a non-natural base containing nucleotide.

[0352] In certain embodiments, at least one modification may be a 2'-O-methyl modification in the ribose moiety.

[0353] In certain embodiments, at least one modification may be a 2'-F modification of the ribose moiety.

[0354] In certain embodiments, the first nucleic acid portion of (i)(a); and / or the second nucleic acid portion of (ii)(b); and / or the nucleotide at any one of positions 2 and 14 downstream of the first nucleotide of the 5' region of (iii), in terms of the range of presence, 1 to 8 additional nucleic acid portions as previously defined herein; may not comprise a 2'-O-methyl modification in the ribose moiety.

[0355] In certain embodiments, the third nucleic acid portion of (i)(c); and / or the fourth nucleic acid portion of (ii)(d); and / or (iii) one, two or all three nucleotides in the passenger nucleic acid portion (if present) (as defined herein before) correspond in position to the first nucleic acid portion of (i)(a); and / or the second nucleic acid portion of (ii)(b); and / or (iii) any nucleotide in positions 11 to 13 downstream of the first nucleotide of the 5' region of the 1 to 8 additional nucleic acid portions (if present), respectively, as defined herein before; may not comprise a 2'-O-methyl modification in the ribose moiety.

[0356] In certain embodiments, nucleotides at any one of positions 2 and 14 downstream of the first nucleic acid portion of (i)(a); and / or the second nucleic acid portion of (ii)(b); and / or the first nucleic acid portion in (iii) may, in terms of the scope present, have 1 to 8 additional nucleic acid portions as previously defined herein; and may comprise a 2'-F modification in the ribose moiety.

[0357] In certain embodiments, the third nucleic acid portion of (i)(c); and / or the fourth nucleic acid portion of (ii)(d); and / or

[0358] (iii) one, two, or all three nucleotides in the (if present) passenger nucleic acid portion (as defined previously herein) may comprise a 2'-F modification in the ribose moiety, the passenger nucleic acid portion corresponding in position to the first nucleic acid portion of (i)(a); and / or the second nucleic acid portion of (ii)(b); and / or (iii) (if present) any nucleotide at positions 11 to 13 downstream of the first nucleotide in the 5' region of the 1 to 8 additional nucleic acid portions, and the passenger nucleic acid portion (as defined previously herein) may comprise a 2'-F modification in the ribose moiety.

[0359] In certain embodiments, all remaining nucleotides may contain a 2'-O-methyl modification or a 2'-F modification in the ribose moiety, preferably, except for unmodified nucleotides, according to the labile linkages defined herein. Preferably, the remaining nucleotides may contain a 2'-O-methyl modification in the ribose moiety.

[0360] In certain embodiments, the one or more, preferably one unmodified nucleotide represents any one of the nucleotides of the nucleic acid linker portion as further defined previously herein, preferably the nucleotides of the nucleic acid linker portion as further defined previously herein, which is adjacent to the third nucleic acid portion of (i)(c); and / or the fourth nucleic acid portion of (ii)(d); and / or (iii) in the scope where present, the passenger nucleic acid portion.

[0361] In certain embodiments,

[0362] (a) the first nucleic acid portion may be selected from Table 3a;

[0363] (b) the second nucleic acid portion may be selected from Table 3a;

[0364] (c) the third nucleic acid portion may be selected from Table 3b; and / or

[0365] (d) the fourth nucleic acid portion may be selected from Table 3b.

[0366] In a preferred embodiment, the first nucleic acid portion and the second nucleic acid portion can be selected from Table 3a, wherein the first and second nucleic acid portions are different; and the third and fourth nucleic acid portions can be selected from Table 3b.

[0367] In certain embodiments, the 3'-terminal positions of the first and the third nucleic acid portions can be substituted with unmodified nucleotides.

[0368] In certain embodiments, the nucleic acid construct can comprise at least one vinyl phosphonate modification, such as in the first nucleic acid portion of (i)(a); and / or in the second nucleic acid portion of (ii)(b); and / or in the 5'-region of (iii), to the extent present, of 1 to 8 additional nucleic acid portions as previously defined herein.

[0369] In certain embodiments,

[0370] the first nucleic acid portion of (a); and / or

[0371] the second nucleic acid portion of (b); and / or

[0372] the third nucleic acid portion of (c); and / or

[0373] the fourth nucleic acid portion of (d); and / or

[0374] to the extent present, of 1 to 8 additional nucleic acid portions as previously defined herein; and / or

[0375] to the extent present, a passenger nucleic acid portion as previously defined herein;

[0376] can be an inverted nucleotide and can be linked to an adjacent nucleotide via the 3'-carbon of the nucleotide and the 3'-carbon of the adjacent nucleotide, and / or can be an inverted nucleotide and can be linked to an adjacent nucleotide via the 5'-carbon of the nucleotide and the 5'-carbon of the adjacent nucleotide.

[0377] In certain embodiments, the inverted nucleotide can be linked to an adjacent nucleotide through a phosphate group via a phosphodiester bond; or can be linked to an adjacent nucleotide through a phosphorothioate group; or can be linked to an adjacent nucleotide through a phosphorodithioate group. Compositions and pharmaceutical groups comprising the shRNA, mxRNA and / or muRNA oligomeric constructs of the present invention Compounds According to a third aspect, the invention relates to a composition comprising an oligomeric compound according to the first aspect of the invention and / or a nucleic acid construct according to the second aspect of the invention, and a physiologically acceptable excipient.

[0378] According to a fourth aspect, the invention relates to a pharmaceutical composition comprising an oligomeric compound according to the first aspect of the invention and / or a nucleic acid construct according to the second aspect of the invention.

[0379] The pharmaceutical composition may further comprise a pharmaceutically acceptable excipient, diluent, antioxidant and / or preservative.

[0380] The oligomeric compound according to the first aspect and / or the construct according to the second aspect may be the sole pharmaceutically active agent. Alternatively, the pharmaceutical composition further comprises one or more other pharmaceutically active agents. The other pharmaceutically active agents are agents that modulate the innate and / or adaptive immune system, such as other oligomeric compounds targeting immune system targets different from complement component C5, preferably interleukin-6; agents that reduce the expression or level of interleukin-6; or agents such as antibodies against complement components, which are preferably eculizumab. Preferably the oligomeric compound and / or the nucleic acid construct; and the other pharmaceutically active agents will be administered simultaneously or in any order.

[0381] Diseases to be treated by the shRNA, mxRNA and / or muRNA oligomeric compounds of the present invention and their further uses According to a fifth aspect, the present invention relates to an oligomeric compound according to the first aspect of the present invention and / or a nucleic acid construct according to the second aspect of the present invention for use in human or veterinary medicine or therapy.

[0382] According to a sixth aspect, the present invention relates to an oligomeric compound according to the first aspect of the present invention and / or a nucleic acid construct according to the second aspect of the present invention for use in a method of treating, ameliorating and / or preventing a disease or disorder.

[0383] The disease or disorder may be a C5-related disease or disorder or a disease or disorder that requires reducing C5 expression.

[0384] Specifically, the disease or disorder is selected from the group consisting of: autoimmune diseases, complement system dysfunction (including abnormal upregulation of complement components (such as C5)), age-related macular degeneration (AMD) (including dry AMD and geographic atrophy), paroxysmal nocturnal hemoglobinuria (PNH), generalized myasthenia gravis (gMG), lupus nephritis (LN), Alzheimer's disease, atherosclerosis, choroid plexus inflammation, atypical hemolytic uremic syndrome (aHUS), C3 glomerulopathy (C3G), Ig-mediated kidney lesions (such as IgA nephropathy and primary membranous nephropathy), asthma, rheumatism, rheumatoid arthritis, systemic lupus erythematosus (SLE), antineutrophil cytoplasmic antibody (ANCA) vasculitis, antiphospholipid antibody syndrome (APS), glomerulonephritis, bullous dermatomyositis pemphigoid, Shiga toxin Escherichia coli-associated hemolytic uremic syndrome, amyotrophic lateral sclerosis (ALS), central nervous system (CNS) diseases, myasthenia gravis (MG), neuromyelitis optica (NMO), dense deposit disease, C3 neuropathy, cold agglutinin disease, humoral and vascular transplant rejection, graft dysfunction, myocardial infarction, asthma, rheumatoid arthritis (RA) sensitization to transplantation, antiphospholipid antibody syndrome; lupus nephritis; ischemia-reperfusion injury; typical or infectious hemolytic uremic syndrome (tHUS); dense deposit disease (DDD); neuromyelitis optica (NO); multifocal motor neuropathy (MMN); multiple sclerosis (MS); macular degeneration (e.g., age-related macular degeneration (AMD); hemolysis, elevated liver enzymes, and low platelets (HELLP) syndrome; thrombotic thrombocytopenic purpura (TTP); spontaneous fetal loss; pauci-immune vasculitis; epidermolysis bullosa; recurrent fetal loss; preeclampsia, traumatic brain injury, myasthenia gravis, cold agglutinin disease, bullous dermatomyositis, Shiga toxin E.co / related hemolytic uremic syndrome, C3 glomerulopathy, antineutrophil cytoplasmic antibody - associated vasculitis, humoral and vascular transplant rejection, graft dysfunction, myocardial infarction, allograft, sepsis, coronary artery disease, dermatomyositis, Graves' disease, atherosclerosis, systemic inflammatory response sepsis, septic shock, spinal cord injury, glomerulonephritis, Hashimoto's thyroiditis, type I diabetes, pemphigus, autoimmune hemolytic anemia (AIHA), ITP, Goodpasture syndrome, Degos disease, antiphospholipid syndrome (APS), catastrophic APS (CAPS), cardiovascular disease, myocarditis, cerebrovascular disease, peripheral vascular disease, renal vascular disease, mesenteric / intestinal vascular disease, vasculitis, Henoch - Schönlein purpura nephritis, systemic lupus erythematosus - related vasculitis, rheumatoid arthritis - related vasculitis, immune complex vasculitis, Takayasu disease, dilated cardiomyopathy, diabetic angiopathy, Kawasaki disease (arteritis), venous gas embolism (VGE), restenosis after stent implantation, rotational atherectomy, membranous nephropathy, Guillain - Barré syndrome and percutaneous transluminal coronary angioplasty age - related macular degeneration (AMD) and / or geographic atrophy (GA); uveitis and / or panuveitis; cold agglutinin disease, membranoproliferative glomerulonephritis (MPGN), Guillain - Barré syndrome, Shiga toxin - producing Escherichia coli hemolytic uremic syndrome (STEC - HUS), organ transplant - related autoimmune diseases and sepsis. According to a seventh aspect, the present invention relates to a method of treating a disease or disorder, comprising administering to an individual in need of treatment an oligomeric compound according to the first aspect and / or a nucleic acid construct according to the second aspect of the present invention.

[0385] Specifically, the disease or disorder is selected from paroxysmal nocturnal hemoglobinuria (PNH), Alzheimer's disease, atherosclerosis, choroid plexus inflammation, generalized myasthenia gravis (gMG), amyotrophic lateral sclerosis (ALS), lupus nephritis (LN), central nervous system (CNS) diseases; age - related macular degeneration (AMD) and / or geographic atrophy (GA); uveitis and / or panuveitis; cold agglutinin disease, membranoproliferative glomerulonephritis (MPGN), Guillain - Barré syndrome, Shiga toxin - producing Escherichia coli hemolytic uremic syndrome (STEC - HUS) and organ transplant - related autoimmune diseases.

[0386] The oligomeric compound and / or nucleic acid construct can be administered to the individual by subcutaneous injection or intravenous injection.

[0387] According to an eighth aspect, the present invention relates to the use of an oligomeric compound according to the first aspect or a nucleic acid construct according to the second aspect for research as a gene function analysis tool.

[0388] According to a ninth aspect, the present invention relates to the use of an oligomeric compound according to the first aspect and / or a nucleic acid construct according to the second aspect in the manufacture of a medicament for treating a disease or disorder.

[0389] Constructs and sequences of the oligomeric compounds of the present invention

[0390] The following table shows the nucleobase sequences of the antisense and sense strands of the oligomeric compounds of the present invention, as well as the nucleobase sequences of the single-stranded oligomeric compounds of the present invention, and the definitions of the modified oligomeric compounds of the present invention (symbols including nucleobase sequences, sugar modifications, and modified phosphates where applicable).

[0391] The symbols used are common in the art and have the following meanings:

[0392] A represents adenine;

[0393] U represents uracil;

[0394] C represents cytosine;

[0395] G represents guanine.

[0396] 5Phos represents a 5'-terminal phosphate group, which is preferred but not essential;

[0397] m represents a methyl modification at the 2'-position of the sugar of the base nucleoside;

[0398] f represents a fluorine modification at the 2'-position of the sugar of the base nucleoside;

[0399] r represents an unmodified (2'-OH) ribonucleotide;

[0400] [Ps] or # represents a phosphorothioate internucleoside bond;

[0401] i represents an inverted internucleoside bonding, which can be 3'-3' or 5'-5';

[0402] 3xGalNAc represents trivalent GalNAc.

[0403] The following Tables 1a and 1b show the nucleobase sequences of the antisense and sense strands of 250 oligomeric compounds according to the examples.

[0404] Table 1a: Nucleobase sequences of the antisense strands of 250 constructs of the present invention

[0405]

[0406]

[0407]

[0408]

[0409]

[0410]

[0411]

[0412] Table 1b: Nucleobase sequences of the sense strands of 250 constructs of the present invention

[0413]

[0414]

[0415]

[0416]

[0417]

[0418]

[0419] The following Table 2 shows the nucleobase sequences of 250 hairpin structures of the present invention selected according to the examples. The nucleobase sequences are the direct fusions of the antisense sequences of Table 1a and the corresponding sense sequences of Table 1b.

[0420] Table 2: Nucleobase sequences of 250 constructs of the present invention, where the sense and antisense sequences of Tables 1a and 1b are combined.

[0421]

[0422]

[0423]

[0424]

[0425]

[0426]

[0427] The following Tables 3a to c show 100 antisense sequences, sense sequences, and hairpin structures of the present invention, respectively; and the complete modification information (modified sugars and modified phosphates when applicable).

[0428] Table 3a: Modified antisense constructs of the present invention

[0429]

[0430]

[0431]

[0432]

[0433]

[0434] Note = Each of the above constructs may or may not have a phosphate modification at the 5'-end group. Additionally, and independently, each of the above constructs may or may not have "3x GalNAc" coupled to the 3'-end group. Constructs with the 3xGalNAc ligand are preferred. Particularly preferred are constructs that additionally have a 5'-phosphate, even though this is not strictly required, as in the absence of phosphate, mammalian cells will add this phosphate in the absence of phosphate in the administered molecule.

[0435] Table 3b: Modified sense constructs of the present invention

[0436]

[0437]

[0438]

[0439]

[0440]

[0441] Note = Each of the above constructs may or may not have "3x GalNAc" coupled to the 3'-end group. Constructs with the 3x GalNAc ligand are preferred, particularly the toothbrush ligand as defined herein.

[0442] Table 3c: Modified hairpin structures of the present invention

[0443]

[0444]

[0445]

[0446]

[0447]

[0448]

[0449]

[0450]

[0451]

[0452] Note = Each of the above constructs may or may not have a phosphate modification at the 5'-end group. Additionally, and independently, each of the above constructs may or may not have "3x GalNAc" coupled to the 3'-end group. Constructs having the 3xGalNAc ligand are preferred, particularly the toothbrush ligand as defined herein. Particularly preferred are constructs that additionally have a 5'-phosphate, even though this is not strictly required since in the absence of phosphate, mammalian cells will add this phosphate in the absence of phosphate in the administered molecule.

[0453] Specific notes on the nomenclature in Tables 3a to 3c:

[0454] fN: 2'-fluoro residue

[0455] mN: 2'-O-methyl residue Postscript: phosphorothioate

[0456] p, Phos: phosphate

[0457] (GalNAc): Sirnaomics mono-GalNAc building block

[0458] It should also be noted that the scope of the present invention extends to sequences corresponding to the sequences in the above table, wherein the 5'-terminal nucleoside of the antisense (guide) strand (the first region as defined in the claims herein) may include any nucleobase that can be present in an RNA molecule, in other words, it can be any one of adenine (A), uracil (U), guanine (G), or cytosine (C). Additionally, the scope of the present invention extends to sequences corresponding to the above table, wherein the 3'-terminal nucleoside of the sense (passenger) strand (the second region as defined in the claims herein) may include any nucleobase that can be present in an RNA molecule, in other words, it can be any one of adenine (A), uracil (U), guanine (G), or cytosine (C), but preferably a nucleobase complementary to the 5'-nucleobase of the antisense (guide) strand (the first region as defined in the claims herein).

[0459] Although these methods are shown and described as a series of acts performed in a particular order, it should be understood and recognized that these methods are not limited by the order of the acts. For example, some acts may occur in a different order than that described herein. Additionally, one act may occur concurrently with another act. Moreover, in certain cases, not all acts may be required to implement the methods described herein.

[0460] The order of steps of the methods described herein is exemplary, but these steps can be performed in any suitable order or simultaneously where appropriate. Additionally, steps can be added or replaced in any method, or individual steps can be deleted from any method without departing from the scope of the subject matter described herein. Aspects of any of the above examples can be combined with aspects of any of the other above examples to form further examples.

[0461] It should be understood that the description of the above preferred embodiments is given by way of example only, and various modifications can be made by those skilled in the art. The above description includes examples of one or more embodiments. Of course, it is not possible to describe all conceivable modifications and changes of the above compounds, compositions or methods for the purpose of describing the above aspects, but those of ordinary skill in the art can recognize that many further modifications and arrangements of various aspects are possible. Accordingly, the aspects are intended to cover all such changes, modifications and variations that fall within the scope of the appended claims. Specific embodiments

[0462] The following examples illustrate certain embodiments of the present disclosure but are not limiting. Additionally, in providing the specific embodiments, the inventors have considered the general application of these specific embodiments. For example, the disclosure of oligonucleotides having a particular motif or modification pattern provides reasonable support for other oligonucleotides having the same or similar motif or modification pattern.

[0463] The synthesis of the RNAi constructs according to the invention and disclosed herein was carried out using synthetic methods known to those skilled in the art, such as the synthetic methods disclosed in https: / / en.wikipedia.org / wiki / Oligonucleo_synthesis {retrieved on February 16, 2022}, wherein the methods disclosed on that website are incorporated herein by reference in their entirety. The only difference from the synthetic methods disclosed in that reference is the use of GalNAc phosphoramidite immobilized on a support during the first step of the synthetic process.

[0464] Example 1

[0465] Materials and methods

[0466] Cell culture:

[0467] Human primary hepatocytes (pooled from 5 donors - Sekisui XenoTech, HPCH05+) were thawed immediately before the experiment and cultured in 1x complete Williams medium (Gibco, A1217601) supplemented with a hepatocyte culture supplement pack (Gibco, CM3000). The FBS concentration was changed from the manufacturing formulation to a final 2.5% (instead of 5%) to ensure compound stability.

[0468] 1x Complete WEM: 2.5% FBS, 1 μM dexamethasone, penicillin / streptomycin (100 U / mL / 100 μg / mL), 4 μg / ml human insulin, 2 mM GlutaMAX, 15 mM HEPES, pH 7.4.

[0469] C5 Target Identification and Duplex Preparation:

[0470] Oligomeric compounds against C5 were identified by bioinformatics analysis of the human C5 mRNA sequence given in RefSeq sequence ID NM_001735.2. 100 compounds were selected for mxRNA hairpin synthesis. The compounds were dissolved in molecular biology grade water to 50 μM. The duplexes were annealed by heating at 95 °C for 5 minutes and then gradually cooled to room temperature. The mxRNA was annealed by heating at 95 °C for 5 minutes and then rapidly cooled on ice.

[0471] C5 - Primary Screening:

[0472] On the day of transfection, primary human hepatocytes were thawed in 45 ml of human OptiThaw (Sekisui XenoTech, K8000) and centrifuged at 200 g for 5 minutes. The cells were suspended in 2x Complete WEM and counted. Then the cells were seeded in 50 μL of 2x Complete WEM in a 96-well type 1 rat tail collagen plate, 25,000 cells per well, and allowed to settle and attach for four hours before transfection. After settling, the compounds were further diluted to 2 μM in basal WEM. 50 μL of each 2 μM compound was added separately to three replicates of the seeded hepatocytes, with a final concentration of 1 μM and a volume of 100 μL of 1x Complete WEM.

[0473] 72 hours after transfection, cells were harvested and RNA was isolated using the PureLink Pro 96 Total RNA Purification Kit (ThermoFisher, 12173011A) according to the manufacturer's protocol. C5 expression of the harvested RNA was detected by Taqman qPCR using the Luna Universal Probe One-Step RT-qPCR Kit (NEB, E3006). Each sample was subjected to qPCR using the C5 TaqMan probe set (Hs01004342_m1-FAM) multiplexed with the universal GAPDH VIC probe (ThermoFisher, 4326317E). Thermal cycling and data acquisition were performed using an Applied Biosystems QuantStudio 3 / 5 real-time PCR system.

[0474] C5 - Secondary Screening:

[0475] Based on the preliminary screening data, a set of 25 C5-targeting mxRNA constructs with the best effects were tested in the dose curve. The compound was further diluted to 2 μM in the basal WEM. A seven-step five-fold dilution series was prepared in the basal WEM, ranging from 2 μM to 0.000128 μM. 50 μL of each dilution was added to three replicates of the inoculated hepatocytes respectively, and the final dilution series was from 1 μM to 0.000064 μM in 100 μL of 1x complete WEM.

[0476] At 72 hours post-transfection, according to the manufacturer's protocol, cells were harvested and RNA was isolated using the PureLink Pro 96 Total RNA Purification Kit (ThermoFisher, 12173011A). C5 expression of the harvested RNA was detected by Taqman qPCR using the Luna Universal Probe One-Step RT-qPCR Kit (NEB, E3006). Each sample was subjected to qPCR detection using the C5 TaqMan probe set (Hs01004342_m1-FAM) multiplexed with the universal GAPDH VIC probe (ThermoFisher, 4326317E). Thermal cycling and data acquisition were performed using the Applied Biosystems QuantStudio 3 / 5 Real-Time PCR System.

[0477] Example 2

[0478] Results

[0479] Figure 1 The preliminary screening results of the selected compounds of the present invention and their activity of inhibiting C5 expression are shown.

[0480] Table 4 below shows the IC50 values (in nM) of 25 preferred constructs selected according to the examples. The maximum % KD represents the maximum knockdown achieved at 1000 nM, where 0% represents no knockdown and 100% represents complete knockdown. M4K4 was used as a reference.

[0481] Construct ID Maximum KD% IC50 C5-m-30 71.88574381 4.943 C5-m-37 77.48131233 11.25 C5-m-83 59.57363723 21.99 C5-m-61 68.93837532 40.04 C5-m-74 63.30023809 41.25 C5-m-82 62.26208156 68.84 C5-m-87 63.87542555 89.55 C5-m-55 63.0068919 90.72 C5-m-23 60.00147173 213.2 C5-m-28 50.59311869 295.4 C5-m-42 49.72928101 302.6 C5-m-73 48.77874599 351.7 C5-m-66 56.30345942 351.9 C5-m-47 53.46576404 367.2 C5-m-46 50.25328616 416.7 C5-m-27 48.71998765 583.3 C5-m-16 50.77488697 629.7 C5-m-43 43.01211645 779.6 C5-m-36 48.1079184 894.2 C5-m-72 44.02312363 1090 C5-m-53 40.3528029 1167 C5-m-14 38.57830035 2282 C5-m-75 38.99786713 3485 C5-m-59 32.76891127 5167 C5-m-56 22.82809496 6117 M4K4 11.680749 3810

[0482] The IC50 data is in the single-digit to low double-digit nanomolar range, demonstrating the excellent performance of numerous constructs of the present invention. In addition, no obvious toxicity was observed.

[0483] The further screening results of the constructs of the present invention in Table 4 above and their excellent performance are as Figure 2 shown.

[0484] Table 5 below shows the IC50 values (in nM) of 6 preferred constructs selected according to the examples. The maximum % KD represents the maximum knockdown achieved at 1000 nM (nanomoles per liter), where 0% represents no knockdown and 100% represents complete knockdown.

[0485] Construct ID KD% at 1000 nM IC50 (nM) C5-30 (large) 85.136214 2.939 C5-37 (large) 85.717658 5.605 C5-83 (large) 71.592597 37.45

[0486] Figure 3 Further results of the constructs in Table 5 above at different concentrations are shown.

[0487] Table 5 and Figure 3 show that the large-scale preparation and screening synthesis of C5 are very close. At the highest dose, the knockdown rates of C5-m-30 and C5-m-37 are 85%, while the knockdown rate of C5-m-83 is 72%.

[0488] Example 3

[0489] Complement component C5-targeted mxRNA leading candidate dose and duration response study in humanized liver-uPA-SCID mouse model, non-GLP Figure 4

[0490] 1. Research Objectives

[0491] The purpose of this non-GLP study is to evaluate the dose and duration responses of GalNAc-conjugated complement component C5-targeted mxRNA constructs in humanized liver-uPA-SCID mice. The compound will be administered subcutaneously and the mice will survive for up to 42 days.

[0492] Plasma and serum will be collected before necropsy. At necropsy, 3 liver biopsy samples (2 mm) from each animal will be separately stored in vials in RNAlater, snap-frozen, and stored at -80 °C. Three additional liver biopsy samples (2 mm) will be collected, snap-frozen, stored in the same vial, and stored at -80 °C.

[0493] 2. Regulatory Compliance

[0494] This non-GLP study will not be conducted in accordance with the regulations of the Food and Drug Administration's Good Laboratory Practice (GLP) (21 CFR Part 58).

[0495] 3. Animal Welfare Compliance

[0496] The protocol has been reviewed and approved by the IACUC committee of the testing facility.

[0497] 4. Test System Information

[0498] 4.1. Animal Experiments

[0499] 4.1.1. Common Name: Mouse

[0500] 4.1.2. Variety / Category: Rodent - Humanized Liver - uPA - SCID Mouse Model

[0501] 4.1.3. Number of Animals (by Gender): 44 males, all neonatal mice

[0502] 4.2. Adaptation Period:

[0503] 4.2.1. Duration:

[0504] All animals will undergo an adaptation period of at least five (5) days before being released, and the attending veterinarian will assess the overall health status of the animals at that time.

[0505] 4.2.2. Required Medications and / or Vaccinations:

[0506] · All received rodents are from certified suppliers free of any lethal parasites that may affect the entire population of the facility.

[0507] · All rodents must be accompanied by sentinel reports including statistical analysis.

[0508] · Each batch of rodents must be housed separately from other animals in the facility.

[0509] 4.3. Animal Identification Method and Location:

[0510] Animals will be assigned consecutive numbers. The supplier will notch the ears before shipping the animals to permanently identify each animal. Animals may have color markings to distinguish similar ear notches. A cage card will also be attached to each animal cage indicating the animal number, gender, supplier, strain, principal investigator, and study number.

[0511] 5. Study Design

[0512] 5.1. Design Details

[0513] This study will use a mouse, N = 44. Animals will be grouped according to treatment type, dose, and survival period. Each animal will be treated with a subcutaneous injection of the test material. For details, please refer to Study Table 1.

[0514] At autopsy, three 2 - millimeter biopsy samples will be taken from the left, middle, and right liver lobes and placed in separate vials, soaked in RNAlater for 15 minutes, flash - frozen, and stored at - 80°C. Another three 2 - millimeter liver biopsy samples taken from the left, middle, and right liver lobes will be placed in one vial, flash - frozen, and stored at - 80°C. The remaining liver will be flash - frozen and stored in a 10 - mL conical tube at - 80°C.

[0515]

[0516] The research plan is also as Results shown.

[0517] Table 6: Dosage Information

[0518]

[0519] Table 7: Research Table

[0520]

[0521] 6. Information on Test Articles and Auxiliary Materials

[0522] 6.1. Test Drug 1:

[0523] 6.1.1. Identification: C5-30 (SEQ ID No.980)

[0524] 6.1.2. Manufacturer: Shennuo Pharmaceutical

[0525] 6.1.3. Description: GalNAc-conjugated human complement component C5-targeted mxRNA

[0526] 6.1.4. Batch Number: Will be recorded on the research data form.

[0527] 6.1.5. Expiry Date: Will be recorded on the research data form.

[0528] 6.1.6. Storage Temperature: 4°C

[0529] 6.1.7. Biological Hazard Status: None

[0530] 6.1.8. Material Safety Data Sheet*: To be determined

[0531] 6.1.9. Appearance: Clear liquid

[0532] 6.1.10. Dosage Information: See Table 6

[0533] 6.1.11. Storage of Residual Test Articles: None

[0534] 6.2. Test Drug 2:

[0535] 6.2.1. Identification: C5-37

[0536] 6.2.2. Manufacturer: Shennuo Pharmaceutical

[0537] 6.2.3. Description: GalNAc-conjugated human complement component C5-targeted mxRNA

[0538] 6.2.4. Batch Number: Will be recorded on the research data form.

[0539] 6.2.5. Cut-off Date: It will be recorded on the research data form.

[0540] 6.2.6. Storage Temperature: 4°C

[0541] 6.2.7. Biological Hazard Status: None

[0542] 6.2.8. Material Safety Data Sheet*: To be determined

[0543] 6.2.9. Appearance: Clear liquid

[0544] 6.2.10. Dosage Information: See Table 6

[0545] 6.2.11. Residual Test Article Storage: None

[0546] Figure 5

[0547] The following table shows the results of knocking down the C5 gene with constructs C5-30 and C5-37 in humanized liver-uPA-SCID mice.

[0548] Table 8a: Results of knocking down (KD) the C5 gene at different doses at several time points with construct C5-30 (structure shown in Table 3c)

[0549]

[0550] Table 8b: Results of knocking down the C5 gene at different doses at several time points with construct C5-37 (structure shown in Table 3c)

[0551]

[0552] The results of the mouse study are also as Figure 6 shown.

[0553] Example 4

[0554] Evaluate the duration effect of mxRNA targeting human complement C5 in a non-GLP model of humanized liver-uPA-SCID mice. This protocol is presented in its original wording. Therefore, any description using the future tense of the verb means that the experiment has been completed.

[0555] 1. Research Objectives

[0556] The objective of this non-GLP study is to evaluate the duration effect of the C5-30 mxRNA compound (see Table 3c; experimental code: C5-m-30 SEQ ID No. 980) targeting human complement C5 mRNA in humanized liver-uPA-SCID mice.

[0557] The compound will be administered by subcutaneous injection, and the mice will survive for up to 84 days.

[0558] 2. Research Design

[0559] 2.1. Design Details

[0560] This study will use one type of mouse, PXB. The animals will be grouped according to treatment type, dose, and survival period. Each animal will be treated by subcutaneous injection of the test material. (Note: The injection must be subcutaneous. If the injection site is incorrect, injected into the muscle area or vein / blood, the test article will not work).

[0561] · Groups 1A, 1B, 1C, and 1D** will each have five animals and receive a single control dose of PBS (phosphate buffered saline).

[0562] · Groups 2A, 2B, 2C, and 2D** will each have five animals and receive a single dose of C5-30 (mxRNA targeting human complement C5 mRNA, SEQ ID No. 980) at a dose of 30 mg / kg.

[0563] The animals will survive for 14 days, 28 days, 56 days, and 84 days respectively. See Figure 7 and Table 9.

[0564] Table 9: Research Table

[0565]

[0566] Note: The structure of C5-30 is shown in Table 3c, and the experimental code is "C5-m-30".

[0567] 3. Test Article and Related Material Information

[0568] 3.1. Test Drug 1:

[0569] 3.1.1. Identification: C5-30

[0570] 3.1.2. Manufacturer: Sirnaomics

[0571] 3.1.3. Description: GalNAc-conjugated human complement component C5-targeting mxRNA

[0572] 3.1.4. Batch Number: Will be recorded in the research material table.

[0573] 3.1.5. Expiration Date: Will be recorded in the research material table.

[0574] 3.1.6. Storage Temperature: 4°C

[0575] 3.1.7. Biohazard Status: None

[0576] 3.1.8. Safety Data Sheet (SDS): To be determined

[0577] 3.1.9. Appearance: Clear liquid

[0578] 3.1.10. Dosage information: See Table 1

[0579] 3.1.11. Storage of remaining test items: None

[0580] Note: The structure of C5-30 is shown in Table 3c, and the experimental code is C5-m-30.

[0581] Results

[0582] The research results are as Figure 7 shown.

[0583] Figure 7 It shows the duration effect of C5 gene expression knockdown (KD):

[0584] - Week 2: C5 mRNA knockdown was 61%;

[0585] - Week 4: C5 mRNA knockdown was 46%;

[0586] - Week 8: C5 mRNA knockdown was 13%;

[0587] - Week 12: Returned to the control group level.

[0588] Note:

[0589] - Two animals died in the control group (at 8 weeks) (remaining N = 4);

[0590] - Two animals died in the C5-30 group (at 4 weeks) (remaining N = 4);

[0591] - Two animals died in the C5-30 group (at 8 weeks) (remaining N = 4).

Claims

1. An oligomeric compound capable of suppressing the expression of complement component C5, wherein the compound comprises at least a first linked nucleoside region, and the base (nucleoside base) sequence of the at least first linked nucleoside region is at least partially complementary to at least a part of the RNA transcribed from the C5 gene, wherein the first nucleoside base sequence is selected from the following sequences or parts thereof: the sequences of Table 1a (SEQ ID NO: 1 to 250), wherein the part preferably has a length of at least 18 nucleosides.

2. The oligomeric compound according to claim 1, which further comprises at least a second linked nucleoside region having at least a second nucleoside base sequence that is at least partially complementary to the first nucleoside base sequence and is selected from the following sequences or parts thereof: the sequences of Table 1b (SEQ ID NO: 251 to 500), wherein the length of the part is preferably at least 8, 9, 10 or 11 nucleosides, more preferably at least 10 nucleosides.

3. The oligomeric compound according to claim 1 or 2, wherein the first nucleoside base sequence is selected from the following sequences or parts thereof: SEQ ID NO: 61, 30, 37, 87, 55, 66, 23, 83, 43, 47, 72, 27, 14, 28, 46, 82, 74, 75, 73, 53, 16, 36, 59, 42 and 56.

4. The oligomeric compound according to claim 3, wherein the second nucleoside base sequence is selected from the following sequences or parts thereof: SEQ ID NO: 311, 280, 287, 337, 305, 316, 273, 333, 293, 297, 322, 277, 264, 278, 296, 332, 324, 325, 323, 303, 266, 286, 309, 292 and 306.

5. The oligomeric compound according to any one of claims 1 to 4, wherein the first nucleoside base sequence is selected from the following sequences or parts thereof: SEQ ID NO: 61, 30, 37, 83, 82, 74, 75, 73, 53, 16, 36, 59, 42 and 56, preferably 30, 37 and 83, more preferably 30 and 37.

6. The oligomeric compound according to claim 5, wherein the second nucleoside base sequence is selected from the following sequences or parts thereof: SEQ ID NO: 311, 280, 287, 333, 332, 324, 325, 323, 303, 266, 286, 309, 292, 306, preferably 280, 287 and 333, more preferably 280 and 287.

7. The oligomeric compound according to any one of claims 1 to 6, wherein the first region of the linked nucleosides consists essentially of 18 to 35, preferably 18 to 20, more preferably 18 or 19, more preferably 19 linked nucleosides.

8. An oligomeric compound according to any one of claims 2 to 7, wherein said second linked nucleoside region consists essentially of 10 to 35, preferably 10 to 20, more preferably 10 to 16, still more preferably 10 to 15, and especially 13, 14 or 15 linked nucleosides.

9. An oligomeric compound according to any one of claims 2 to 8, which comprises at least one complementary duplex region, said complementary duplex region comprising at least a portion of said first linked nucleoside region directly or indirectly linked to at least a portion of said second linked nucleoside region, wherein preferably the length of said duplex region is 10 to 19 base pairs, more preferably 12 to 19 base pairs, and still more preferably 12 to 15 base pairs, especially 14 or 15 base pairs, wherein optionally there is one mismatch within said duplex region.

10. An oligomeric compound according to claim 9, wherein each of said first and second regions of linked nucleosides has a 5' to 3' directionality, thereby defining its 5' and 3' regions respectively.

11. An oligomeric compound according to claim 10, wherein the 5' region of said first linked nucleoside region is directly or indirectly linked to the 3' region of said second linked nucleoside region, for example by complementary base pairing, wherein preferably the 5'-terminal nucleoside of said first nucleoside region base pairs with the 3'-terminal nucleoside of said second nucleoside region.

12. An oligomeric compound according to claim 10 or 11, wherein the 3' region of said first linked nucleoside region is directly or indirectly linked to the 5' region of said second linked nucleoside region, wherein preferably said first nucleoside region is directly covalently linked to said second nucleoside region, for example by a phosphate, phosphorothioate or dithiophosphate, and more preferably, the 3'-terminal nucleoside of said first linked nucleoside region is directly covalently linked to the 5'-terminal nucleoside of said second linked nucleoside region by a phosphate, phosphorothioate or dithiophosphate.

13. An oligomeric compound according to any one of claims 1 to 12, which further comprises one or more ligands.

14. An oligomeric compound according to claim 13, wherein said one or more ligands, especially two or more or three ligands, are conjugated to said second linked nucleoside region and / or said first linked nucleoside region.

15. An oligomeric compound according to claim 14, dependent on claim 10, wherein said one or more ligands are conjugated at the 3' region, preferably at the 3'-terminal nucleoside of the second linked nucleoside region and / or the first linked nucleoside region, and / or conjugated to the 5'-terminal nucleoside of said second linked nucleoside region.

16. An oligomeric compound according to any one of claims 13 to 15, wherein said one or more ligands are any cell-targeting moieties, such as lipids, carbohydrates, aptamers, vitamins and / or peptides that bind to specific targets on the cell membrane or cell surface.

17. An oligomeric compound according to claim 16, wherein said one or more ligands comprise one or more carbohydrates.

18. The oligomeric compound according to claim 17, wherein the one or more carbohydrates can be monosaccharides, disaccharides, trisaccharides, tetrasaccharides, oligosaccharides or polysaccharides.

19. The oligomeric compound according to claim 18, wherein the one or more carbohydrates comprise or consist of one or more hexose moieties.

20. The oligomeric compound according to claim 19, wherein the one or more hexose moieties are one or more galactose moieties, one or more lactose moieties, one or more N-acetylgalactosamine moieties, and / or one or more mannose moieties.

21. The oligomeric compound according to claim 20, wherein the one or more carbohydrates comprise one or more N-acetylgalactosamine moieties.

22. The oligomeric compound according to claim 21, which comprises two or more N-acetylgalactosamine moieties, preferably three.

23. The oligomeric compound according to any one of claims 13 to 22, wherein the one or more ligands are attached to the oligomeric compound in a linear configuration or a branched configuration, preferably to the second region of the linked nucleoside.

24. The oligomeric compound according to claim 23, wherein the one or more ligands are attached to the oligomeric compound in a bi-antennary or tri-antennary structure.

25. The oligomeric compound according to any one of claims 1 to 24, wherein the compound consists of the first linked nucleoside region and the second linked nucleoside region.

26. The oligomeric compound according to any one of claims 1 to 24, wherein there is a third region of linked nucleosides intervening between the first region and the second region.

27. The oligomeric compound according to claim 26, wherein the oligomeric compound comprises or consists of a single strand comprising the first, the third and the second nucleoside regions, wherein at least a portion of the first nucleoside region is directly or indirectly linked to at least a portion of the second nucleoside region, thereby forming the at least partially complementary double-stranded region.

28. The oligomeric compound according to any one of claims 9 to 25, wherein the oligomeric compound comprises or consists of a single strand comprising the first and second linked nucleoside regions, wherein at least a portion of the first linked nucleoside region is directly or indirectly linked to at least a portion of the second linked nucleoside region, thereby forming the at least partially complementary double-stranded region.

29. The oligomeric compound according to claim 28, wherein the first and the second nucleoside regions are directly adjacent on the single strand.

30. The oligomeric compound according to claim 28 or 29, wherein the first nucleoside region has a greater number of linked nucleosides compared to the second nucleoside region, wherein optionally the ratio of the total number of linked nucleosides in the first nucleoside region to the total number of linked nucleosides in the second nucleoside region ranges from about 19 / 15 to about 19 / 8, or from about 18 / 15 to about 18 / 8; and / or the percentage of the total number of nucleosides linked to the first nucleoside region relative to the total number of nucleosides of the oligomeric compound ranges from about 55% to about 60%.

31. The oligomeric compound of claim 30, wherein an additional number of linking nucleosides in the first nucleoside region form a hairpin loop that links the first and second linking nucleoside regions, wherein preferably a part of the first nucleoside base sequence of the first nucleoside base sequence is complementary RNA transcribed from the C5 gene to form the hairpin loop, and wherein the loop contains 2 to 5, preferably 4 or 5 nucleosides.

32. The oligomeric compound according to any one of claims 27 to 31, wherein the single strand has a nucleoside base sequence selected from Table 2, in particular SEQ ID NO: 561, 530, 537, 587, 555, 566, 523, 583, 543, 547, 572, 527, 514, 528, 546, 582, 574, 575, 573, 553, 516, 536, 559, 542, and 556, preferably 583, 530, and 537, more preferably 530 and 537.

33. The oligomeric compound according to claim 32, wherein the single strand is selected from Table 3c, in particular selected from construct ID NO: 1011, 980, 987, 1037, 1005, 1016, 973, 1033, 993, 997, 1022, 977, 964, 978, 996, 1032, 1024, 1025, 1023, 1003, 966, 986, 1009, 992, 1006, preferably 980, 987, and 1033, more preferably 980 and 987.

34. The oligomeric compound according to claim 33, subordinate to claim 10, wherein the hairpin loop is present in the 3' region of the first linking nucleoside region, and wherein optionally, one, two, or more 3'-terminal nucleosides of the first nucleoside base sequence, within the range permitted by the nucleoside bases of the one, two, or more 3'-terminal nucleosides, fold back and form or contribute to the second linking nucleoside region.

35. The oligomeric compound of claim 26 or 27, wherein the third nucleoside region and optionally the 3'-terminal portion of the first nucleoside region (preferably consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 linked nucleosides) and / or the 5'-terminal portion of the second nucleoside region (preferably consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 linked nucleosides) form a hairpin loop.

36. The oligomeric compound according to any one of claims 29 to 31, wherein the hairpin loop contains 1 to 8, 2 to 7 linked nucleosides, 3 to 6, preferably 4 or 5 linked nucleosides.

37. The oligomeric compound according to any one of claims 1 to 36, which contains internucleoside linkages and wherein at least one internucleoside linkage is a modified internucleoside linkage.

38. The oligomeric compound according to claim 37, wherein the modified internucleoside bond is a phosphorothioate or dithiophosphate internucleoside bond.

39. The oligomeric compound according to claim 38, which contains 1 to 16 phosphorothioate or dithiophosphate internucleoside bonds.

40. The oligomeric compound according to claim 39, which comprises 7, 8, 9 or 10 phosphorothioate or dithiophosphate internucleoside linkages.

41. The oligomeric compound according to any one of claims 38 to 40, which depends on claim 10 and comprises one or more phosphorothioate or dithiophosphate internucleoside linkages at the 5' region of the first region connecting the nucleosides.

42. The oligomeric compound according to any one of claims 38 to 41, which depends on claim 10 and comprises one or more phosphorothioate or dithiophosphate internucleoside linkages at the 5' region of the second nucleoside connecting region, wherein preferably, the oligomeric compound comprises three phosphorothioate internucleoside linkages at three adjacent nucleosides in the 5' region.

43. The oligomeric compound according to any one of claims 38 to 42, which depends on any one of claims 30 to 32 and comprises phosphorothioate or dithiophosphate internucleoside linkages between at least two, preferably at least three, preferably at least four, preferably at least five adjacent nucleosides of the hairpin loop, depending on the number of nucleosides present in the hairpin loop.

44. The oligomeric compound according to claim 43, which comprises phosphorothioate or dithiophosphate internucleoside linkages between each adjacent nucleoside present in the hairpin loop.

45. The oligomeric compound according to any one of claims 1 to 44, wherein at least one nucleoside comprises a modified sugar.

46. The oligomeric compound according to claim 45, wherein the modified sugar is selected from 2'-modified sugars, conformationally restricted nucleoside (CRN) sugars such as locked nucleic acid (LNA) sugars, (S)-constrained ethyl bicyclic nucleic acid and constrained ethyl (cEt) sugars, tricyclic DNA, morpholine, unlocked nucleic acid (UNA) sugars, glycol nucleic acid (GNA), D-hexitol nucleic acid (HNA) and cyclohexene nucleic acid (CeNA).

47. The oligomeric compound according to claim 46, wherein the 2'-modified sugar is selected from 2'-O-alkyl modified sugars, 2'-O-methyl modified sugars, 2'-O-methoxyethyl modified sugars, 2'-O-allyl modified sugars, 2'-C-allyl modified sugars, 2'-deoxy modified sugars such as 2'-deoxyribose, 2'-F modified sugars, 2'-arabino-fluoride modified sugars, 2'-O-benzyl modified sugars and 2'-O-methyl-4-pyridyl modified sugars.

48. The oligomeric compound according to claim 47, wherein at least one modified sugar is a 2'-O-methyl modified sugar.

49. The oligomeric compound according to claim 47 or 48, wherein at least one modified sugar is a 2'-F modified sugar, and preferably, at most 16 or 17 sugars are 2'-F modified sugars.

50. The oligomeric compound of claim 48 or 49, wherein the sugar is ribose.

51. The oligomeric compound according to any one of claims 48 to 50, which depends on claim 10, wherein the sugar of the nucleoside at either position 2 or 14 downstream of the first nucleoside in the 5' region of the first region connecting the nucleosides does not contain a 2'-O-methyl modification.

52. The oligomeric compound according to any one of claims 48 to 51, wherein the 3'-terminal position of the second region connecting the nucleosides does not contain a 2'-O-methyl modification.

53. The oligomeric compound according to any one of claims 48 to 52, wherein the sugar of the nucleoside at any one of positions 2 and 14 downstream of the first nucleoside in the 5'-region of the first region connecting the nucleosides contains a 2'-F modification.

54. The oligomeric compound according to any one of claims 52 to 53, wherein the sugar of the nucleoside in the second region connecting the nucleosides contains a 2'-F modification, the sugar of the nucleoside in the second region connecting the nucleosides corresponds in position to any one of the nucleosides at positions 11 to 13 downstream of the first nucleoside in the 5'-region of the first region connecting the nucleosides, and the sugar of the nucleoside in the second region connecting the nucleosides contains a 2'-F modification.

55. The oligomeric compound of claim 53 or 54, wherein the 3'-terminal nucleoside of the second region connecting the nucleosides contains a 2'-F modification.

56. The oligomeric compound according to any one of claims 52 to 55, as dependent on claim 10, wherein one or more odd-numbered nucleosides starting from the 5'-region of the first region connecting the nucleosides are modified, and / or wherein one or more even-numbered nucleosides starting from the 5'-region of the first region connecting the nucleosides are modified, wherein the modification of the even-numbered nucleosides is generally a second modification different from the modification of the odd-numbered nucleosides.

57. The oligomeric compound according to claim 56, wherein one or more odd-numbered nucleosides starting from the 3'-region of the second region connecting the nucleosides are modified with a modification different from the modification of the odd-numbered nucleosides in the first region connecting the nucleosides.

58. The oligomeric compound according to claim 56 or 57, wherein one or more of the even-numbered nucleosides starting from the 3'-region of the second region connecting the nucleosides are modified by a modification different from the modification of the even-numbered nucleosides in the first region connecting the nucleosides according to claim 51.

59. The oligomeric compound according to any one of claims 56 to 58, wherein at least one or more modified even-numbered nucleosides in the first region connecting the nucleosides are adjacent to at least one or more differently modified odd-numbered nucleosides in the first nucleoside region.

60. The oligomeric compound according to any one of claims 56 to 59, wherein at least one or more modified even-numbered nucleosides in the second nucleoside region are adjacent to at least one or more differently modified odd-numbered nucleosides in the second region connecting the nucleosides.

61. The oligomeric compound according to any one of claims 56 to 60, wherein the sugar of one or more of the odd-numbered nucleosides starting from the 5'-region of the first nucleoside region is a 2'-O-methyl modified sugar.

62. The oligomeric compound according to any one of claims 56 to 61, wherein one or more of the even-numbered nucleosides starting from the 3'-region of the first region connecting the nucleosides are 2'-F modified sugars.

63. The oligomeric compound according to any one of claims 56 to 62, wherein the sugar of one or more of the odd-numbered nucleosides starting from the 5'-region of the second region connecting the nucleosides is a 2'-O-methyl modified sugar.

64. The oligomeric compound according to any one of claims 56 to 63, wherein one or more of the even-numbered nucleosides starting from the 5'-region of the second linked nucleoside region are 2'-F-modified sugars.

65. The oligomeric compound according to any one of claims 45 to 64, wherein the sugars of a plurality of adjacent nucleosides in the first nucleoside region are modified by common or different modifications.

66. The oligomeric compound according to any one of claims 45 to 65, wherein the sugars of a plurality of adjacent nucleosides in the second nucleoside region are modified by common or different modifications.

67. The oligomeric compound according to any one of claims 56 to 66, subordinated to any one of claims 30 to 33, wherein the sugars of a plurality of adjacent nucleosides in the hairpin loop are modified by common or different modifications.

68. The oligomeric compound according to any one of claims 65 to 67, wherein the common modification is a 2'-F-modified sugar.

69. The oligomeric compound according to any one of claims 65 to 67, wherein the common modification is a 2'-O-methyl-modified sugar.

70. The oligomeric compound according to claim 69, wherein the plurality of adjacent 2'-O-methyl-modified sugars are present in at least eight adjacent nucleosides in the first and / or second nucleoside regions.

71. The oligomeric compound according to claim 70, wherein the plurality of adjacent 2'-O-methyl-modified sugars are present in three or four adjacent nucleosides in the hairpin loop.

72. The oligomeric compound according to claim 46, subordinated to any one of claims 31 to 35, wherein the hairpin loop contains at least one nucleoside having a modified sugar.

73. The oligomeric compound according to claim 72, wherein the at least one nucleoside is adjacent to a nucleoside having a sugar with a different modification, and preferably all adjacent nucleosides in the hairpin loop have sugars with different modifications.

74. The oligomeric compound according to claim 73, wherein the modified sugar is a 2'-O-methyl-modified sugar, and the sugar with a different modification is a 2'-F-modified sugar.

75. The oligomeric compound according to any one of claims 1 to 74, wherein one or more nucleosides in the first linked nucleoside region and / or the second linked nucleoside region are inverted nucleosides and are linked to adjacent nucleosides via the 3'-carbon of their sugar and the 3'-carbon of the sugar of the adjacent nucleoside, and / or one or more nucleosides in the first linked nucleoside region and / or the second linked nucleoside region are inverted nucleosides and are linked to adjacent nucleosides via the 5'-carbon of their sugar and the 5'-carbon of the sugar of the adjacent nucleoside.

76. The oligomeric compound according to any one of claims 1 to 75, which is blunt-ended.

77. The oligomeric compound according to any one of claims 1 to 76, wherein the first or second nucleoside region has an overhang.

78. The oligomeric compound according to any one of claims 1 to 77, wherein the first region is selected from the sequences of Table 3a or portions thereof, particularly selected from construct ID NO: 811, 780, 787, 837, 805, 816, 773, 833, 793, 797, 822, 777, 764, 778, 796, 832, 824, 825, 823, 803, 766, 786, 809, 792, and 806, preferably 780, 787, and 833, more preferably 780 and 787.

79. The oligomeric compound according to any one of claims 1 to 79, wherein said second region is selected from the sequences of Table 3b, or a portion thereof, in particular a portion of 14 nucleotides in length, especially from construct ID NO: 911, 880, 887, 937, 905, 916, 873, 933, 893, 897, 922, 877, 864, 878, 896, 932, 924, 925, 923, 903, 866, 886, 909, 892, 906, preferably 880, 887, 933, more preferably 880 and 887.

80. The oligomeric compound according to any one of claims 1 to 80, wherein the total length of said oligomeric compound is from about 25 to about 35 nucleotides, in particular about 33 or about 34 nucleotides.

81. The oligomeric compound according to any one of claims 10 to 80, wherein the terminal nucleotide at the 5'-position of said first region has a nucleobase selected from A, U, G, and C, preferably U, and wherein optionally, the terminal nucleotide at the 3'-position of said second region has a base complementary to the base at the 5'-position of said first region, preferably A.

82. A nucleic acid construct, comprising at least: (a) a first nucleic acid portion that is at least partially complementary to at least a first portion of the RNA transcribed from the C5 gene; (b) a second nucleic acid portion that is at least partially complementary to at least a second portion of the RNA transcribed from the C5 gene, said second portion being different from the first portion; (c) a third nucleic acid portion that is at least partially complementary to said first nucleic acid portion of (a), thereby forming a first nucleic acid duplex region therewith; (d) a fourth nucleic acid portion that is at least partially complementary to said second nucleic acid portion of (b), thereby forming a second nucleic acid duplex region therewith.

83. The construct according to claim 82, wherein the construct is designed such that after in vivo administration, the construct dissociates to produce at least first and second phase-separated nucleic acid targeting molecules that respectively target the RNA portions transcribed from the target genes of (a) and (b); wherein (i) said first nucleic acid targeting molecule is capable of regulating the expression of the target gene of (a) and comprises or is derived from at least said first nucleic acid portion of (a), and (ii) said second nucleic acid targeting molecule is capable of regulating the expression of the target gene of (b) and comprises or is derived from said second nucleic acid portion of (b).

84. The construct according to claim 82 or 83, wherein the construct is designed to be dissociable such that said first and second phase-separated (discrete) nucleic acid targeting molecules are respectively processed by independent RNAi-induced silencing complexes.

85. A construct according to any one of claims 82 to 84, further comprising at least one labile functional group such that the construct is cleaved after in vivo administration to produce the at least first and second discrete nucleic acid targeting molecules.

86. The construct according to claim 85, wherein the labile functionality comprises one or more unmodified nucleotides.

87. The construct according to claim 86, wherein the one or more unmodified nucleotides of the labile functionality represent one or more cleavage positions within the construct such that after in vivo administration, the construct is cleaved at the one or more cleavage positions to produce the at least first and second phase-separated nucleic acid targeting molecules.

88. The construct according to claim 87, wherein the cleavage positions are located within the construct such that after cleavage, the first discrete nucleic acid targeting molecule comprises or is derived from the first nucleic acid duplex region and the second discrete nucleic acid targeting molecule comprises or is derived from the second nucleic acid duplex region.

89. The construct according to claim 88, wherein the first discrete nucleic acid targeting molecule comprises or consists of the first nucleic acid portion of (a) and the third nucleic acid portion of (c), and / or the second discrete nucleic acid targeting molecule comprises or consists of the second nucleic acid portion of (b) and the fourth nucleic acid portion of (d).

90. The construct according to any one of claims 82 to 89, wherein (a) the first nucleic acid portion has a nucleobase sequence selected from SEQ ID NOs: 1 to 250 in Table 1a; (b) the second nucleic acid portion has a nucleobase sequence selected from Table 1a (SEQ ID NOs: 1 to 250); (c) the third nucleic acid portion has a nucleobase sequence selected from SEQ ID NOs: 251 to 500 in Table 1b; and / or (d) the fourth nucleic acid portion has a nucleobase sequence selected from Table 1b (SEQ ID NOs: 251 to 500). Wherein the third and fourth nucleobase sequences may be one, two or three nucleobases shorter within the range of their 14-nucleobase length, with the 5'-terminal nucleobase preferably absent.

91. The construct according to any one of claims 82 to 90, wherein the first nucleic acid portion of (a) is directly or indirectly linked to the fourth nucleic acid portion of (d) as a primary structure.

92. The construct according to claim 91, wherein the first and fourth nucleic acid portions have nucleobase sequences of SEQ ID NOs: 30 and 287, 30 and 333, 37 and 280, 37 and 333, 83 and 280, 83 and 287, and preferably, the sequences of SEQ ID NOs: 280, 287 and 333 may be one, two, three or four nucleobases shorter, with the 5'-terminal nucleobase preferably absent.

93. A construct according to any one of claims 82 to 92, wherein the second nucleic acid moiety of (b) is directly or indirectly linked to the third nucleic acid moiety of (c) as a primary structure.

94. A construct according to claim 92 or 93, wherein the second and third nucleic acid moieties have nucleobase sequences of SEQ ID NO: 30 and 287, 30 and 333, 37 and 280, 37 and 333, 83 and 280, 83 and 287, respectively, and preferably, the sequences of SEQ ID NO: 280, 287, and 333 may be one, two, three, or four nucleobases shorter, with the 5'-terminal nucleobase preferably absent.

95. A construct according to any one of claims 82 to 90, 91, or 93, further comprising 1 to 8 additional nucleic acid moieties, each of the additional nucleic acid moieties being at least partially complementary to an additional 1 to 8 RNA moieties transcribed from one or more target genes, the target genes may be the same or different from each other, and / or the same or different from the target genes defined in (a) and / or (b), and wherein each of the 1 to 8 additional nucleic acid moieties forms an additional duplex region with a respective passenger nucleic acid moiety, the passenger nucleic acid moiety being at least partially complementary to it.

96. A construct according to claim 95, wherein the second nucleic acid moiety of (b) and the 1 to 8 additional nucleic acid moieties are directly or indirectly linked to a selected passenger nucleic acid moiety as their respective primary structures.

97. A construct according to any one of claims 91, 93, or 96, wherein the direct or indirect linkage represents (i) an internucleotide bond, (ii) an internucleotide nick, or (iii) a nucleic acid linker moiety of 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 nucleotides, the nucleic acid linker being preferably single-stranded.

98. A construct according to claim 97(i), wherein the linkage is direct, resulting in a continuous strand.

99. A construct according to any one of claims 82 to 99, especially claim 97(i), wherein there is a certain complementarity between the first nucleic acid moiety of (a) and the second nucleic acid moiety of (b), or between the third nucleic acid moiety of (c) and the fourth nucleic acid moiety of (d).

100. A construct according to claim 99, wherein the complementarity (i) is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 base pairs, preferably 2, 3, 4, or 5 base pairs; and / or (ii) is located between the first nucleic acid moiety of (a) and the second nucleic acid moiety of (b).

101. A construct according to claims 97(i) to 100, subordinate to claim 86, wherein the internucleotide bond involves at least one of the one or more unmodified nucleotides, and preferably cleavage occurs at the 3'-position of (at least one) the unmodified nucleotide.

102. The construct according to any one of claims 82 to 101, wherein the length of the first nucleic acid moiety of (a), and / or the second nucleic acid moiety of (b), and / or the third nucleic acid moiety of (c), and / or the fourth nucleic acid moiety of (d) is 7 to 25 nucleotides, respectively.

103. The construct according to claim 102, wherein the length of the first nucleic acid moiety of (a) and / or the second nucleic acid moiety of (b) is 18 to 21 nucleotides, more preferably 18 to 20 nucleotides, more preferably 19 nucleotides.

104. The construct according to claim 102 or 103, wherein the length of the third nucleic acid moiety of (c), and / or the fourth nucleic acid moiety of (d) is 11 to 20, more preferably 13 to 16, more preferably 14 or 15, and most preferably 14 nucleotides.

105. The construct according to any one of claims 102 to 104, wherein the unmodified nucleotide is located at any one of positions 18 to 25, more preferably at any one of positions 18 to 21, and / or at the 3'-terminal position of the first nucleic acid moiety of (a) and / or the third nucleic acid moiety of (c).

106. The construct according to claim 105, wherein the unmodified nucleotide is located at position 19.

107. The construct according to any one of dependent claims 99 to 100 or 102 to 105 of claim 87(iii), wherein the length of the nucleic acid linker moiety is 1 to 8 nucleotides, preferably a length of 2 to 7 or 3 to 6 nucleotides, more preferably a length of about 4 or 5 and most preferably a length of 4 nucleotides.

108. The construct according to any one of claims 103 to 107, wherein one or more of all duplex regions independently have a length of 10 to 19 base pairs, more preferably 13 to 19 base pairs, still more preferably 13, 14 or 15 base pairs, and most preferably 14 base pairs, wherein optionally, there is one mismatch within the duplex region.

109. The construct according to any one of claims 82 to 108, which further comprises one or more ligands.

110. The construct according to any one of claims 82 to 109, wherein the first nucleic acid moiety of (a), and / or the second nucleic acid moiety of (b), and / or the third nucleic acid moiety of (c), and / or the fourth nucleic acid moiety of (d), and / or (to the extent present) the 1 to 8 additional nucleic acid moieties as defined in claims 14 and 15, and / or the passenger nucleic acid moiety as defined in claim 95 or 96, respectively have 5'-to-3' directionality, thereby defining their 5' and 3' regions.

111. The construct according to any one of claims 109 or 110, wherein one or more ligands are conjugated to (i) the third nucleic acid moiety of (c), and / or (ii) the fourth nucleic acid moiety of (d), and / or, to the extent present, the 3'-region, preferably the 3'-end, of the passenger nucleic acid moiety as defined in claim 95 or 96.

112. A construct according to any one of claims 109 to 111, wherein one or more ligands are conjugated to one or more regions intermediate the 5' and 3' regions of any of said nucleic acid moieties, preferably the third nucleic acid moiety of (c), and / or the fourth nucleic acid moiety of (d), and / or the passenger nucleic acid moiety defined in claim 95 or 96.

113. A construct according to any one of claims 109 to 112, wherein one or more ligands are conjugated to the 5' region of any of said nucleic acid moieties, preferably the 5' end.

114. A construct according to any one of claims 109 to 113, wherein said one or more ligands are any cell targeting moieties, such as lipids, carbohydrates, aptamers, vitamins, and / or peptides that bind to specific targets on the cell membrane or cell surface.

115. A construct according to claim 114, wherein said one or more carbohydrates can be monosaccharides, disaccharides, trisaccharides, tetrasaccharides, oligosaccharides, or polysaccharides.

116. A construct according to claim 115, wherein said one or more carbohydrates comprise one or more hexose moieties.

117. A construct according to claim 116, wherein said one or more hexose moieties are one or more galactose moieties, one or more lactose moieties, one or more N-acetylgalactosamine moieties, and / or one or more mannose moieties.

118. A construct according to claim 117, which comprises two or three N-acetylgalactosamine moieties.

119. A construct according to any one of claims 109 to 118, wherein said one or more ligands are linked in a linear configuration or a branched configuration.

120. A construct according to claim 119, wherein said one or more ligands are linked in a bi-antennary or tri-antennary structure, or in a structure based on a single ligand at different positions.

121. A construct according to claim 118 or 119, wherein the ligand has the following structure:

122. A construct according to any one of claims 82 to 121, which further comprises one or more phosphorothioate or phosphorodithioate internucleotide linkages.

123. A construct according to claim 122, which comprises 1 to 15 phosphorothioate or phosphorodithioate internucleotide linkages.

124. A construct according to claim 122 or 123, which comprises one or more phosphorothioate or phosphorodithioate internucleotide linkages at one or more 5' and / or 3' regions of the first nucleic acid moiety of (a), and / or the second nucleic acid moiety of (b), and / or the third nucleic acid moiety of (c), and / or the fourth nucleic acid moiety of (d), and / or the one or more additional nucleic acid moieties defined in claim 95 or 96, and / or the passenger nucleic acid moiety defined in claim 95 or 96.

125. A construct according to any one of claims 122 to 124, which comprises a phosphorothioate or phosphorodithioate internucleotide linkage between at least two adjacent nucleotides of the nucleic acid linker moiety defined in claim 97(iii).

126. A construct according to any one of claims 125, comprising phosphorothioate or phosphorodithioate internucleotide linkages between each adjacent nucleotide present in said nucleic acid linker portion.

127. A construct according to any one of claims 122 to 126, comprising phosphorothioate or phosphorodithioate internucleotide linkages that connect: (a) a first nucleic acid portion that is a nucleic acid linker portion as described in claim 97(iii); and / or (b) a second nucleic acid portion that is a nucleic acid linker portion as defined in claim 97(iii); and / or (c) a third nucleic acid portion that is a nucleic acid linker portion as described in claim 97(iii), and / or (d) a fourth nucleic acid portion that is a nucleic acid linker portion as defined in claim 97(iii); and / or one to eight additional nucleic acid portions as defined in claims 95 or 96 to a nucleic acid linker portion as defined in claim 97(iii); and / or a passenger nucleic acid portion as defined in claims 95 or 96 to a nucleic acid linker portion as defined in claim 97(iii).

128. A construct according to any one of claims 82 to 127, wherein at least one nucleotide of at least one of the following is modified: (a) the first nucleic acid portion; and / or (b) the second nucleic acid portion; and / or (c) the third nucleic acid portion; and / or (d) the fourth nucleic acid portion; and / or to the extent present, one to eight additional nucleic acid portions as defined in claims 95 or 96; and / or to the extent present, a passenger nucleic acid portion as defined in claims 95 or 96; and / or to the extent present, a nucleic acid linker portion as defined in claim 97(iii).

129. A construct according to claim 128, wherein one or more odd-numbered nucleotides starting from the 5' region of one of the following are modified, and / or wherein one or more even-numbered nucleotides starting from the 5' region of one of the following are modified, wherein the modification of even-numbered nucleotides is typically a second modification different from the modification of odd-numbered nucleotides: (a) the first nucleic acid portion; and / or (b) the second nucleic acid portion; and / or (c) the third nucleic acid portion; and / or (d) the fourth nucleic acid portion; and / or to the extent present, one to eight additional nucleic acid portions as defined in claims 95 or 96; and / or to the extent present, a passenger nucleic acid portion as defined in claims 95 or 96.

130. A construct according to claim 128 or 129, wherein one or more of the odd-numbered nucleotides starting from the 3' region of the third nucleic acid portion of (c) are modified by a modification different from the modification of the odd-numbered nucleotides starting from the 5' region of the first nucleic acid portion of (a); and / or Among them, one or more of the odd-numbered nucleotides starting from the 3’ region of the fourth nucleic acid portion of (d) are modified, said modification being different from the modification of the odd-numbered nucleotides starting from the 5’ region of the second nucleic acid portion of (b); and / or One or more odd-numbered nucleotides starting from the 3' region of the passenger nucleic acid moiety as defined in claim 95 or 96, to the extent present, are modified by a modification different from the modification of the odd-numbered nucleotides starting from the 5' region of one to eight additional nucleic acid moieties as defined in claim 95 or 96; and / or One or more nucleotides of the nucleic acid linker moiety as defined in claim 97(iii), to the extent present, are modified by a modification that (i) is different from the modification of the adjacent nucleotide in the 3' region of the first nucleic acid moiety of (a); and / or (ii) is different from the modification of the adjacent nucleotide in the 3' region of the second nucleic acid moiety of (b); and / or is different from the modification of the adjacent nucleotide in the 3' region of one to eight additional nucleic acid moieties, to the extent present, as defined in claim 95 or 96.

131. The construct according to any one of claims 128 to 130, wherein: (i) The third nucleic acid moiety of (c), and / or (ii) the fourth nucleic acid moiety of (d), and / or (iii) one or more of the even-numbered nucleotides starting from the 3' region of the passenger nucleic acid moiety as defined in claim 95 or 96, to the extent present, are modified by a modification different from the modification of the odd-numbered nucleotides starting from the 3' region of these respective moieties.

132. The construct according to any one of claims 128 to 131, wherein (i) the first nucleic acid moiety of (a), and / or (ii) the second nucleic acid moiety of (b), and / or (iii) at least one or more of the modified even-numbered nucleotides, to the extent present, are adjacent to at least one or more odd-numbered nucleotides that are differently modified from these respective moieties according to one to eight additional nucleic acid moieties as defined in claim 95 or 96.

133. The construct according to any one of claims 128 to 132, wherein (i) the third nucleic acid moiety of (c), and / or (ii) the fourth nucleic acid moiety of (d), and / or (iii) at least one or more of the modified even-numbered nucleotides, to the extent present, are adjacent to at least one or more odd-numbered nucleotides that are differently modified from these respective moieties according to the passenger nucleic acid moiety as defined in claim 95 or 96.

134. The construct according to any one of claims 128 to 133, wherein (i) the first nucleic acid moiety of (a), and / or (ii) the second nucleic acid moiety of (b), and / or (iii) a plurality of adjacent nucleotides, to the extent present, are modified by a common modification according to one to eight additional nucleic acid moieties as defined in claim 95 or 96.

135. The construct according to any one of claims 128 to 134, wherein (i) the third nucleic acid moiety of (c), and / or (ii) the fourth nucleic acid moiety of (d), and / or (iii) a plurality of adjacent nucleotides, to the extent present, are modified by a common modification according to the passenger nucleic acid moiety as defined in claim 95 or 96.

136. A construct according to claim 134 or 135, wherein the plurality of adjacent co-modified nucleotides are 2 to 4 adjacent nucleotides, preferably 3 or 4 adjacent nucleotides.

137. A construct according to claim 136, wherein the plurality of adjacent co-modified nucleotides are located in (i) the third nucleic acid portion of (c), and / or (ii) the fourth nucleic acid portion of (d), and / or (iii) within the 5' region, to the extent present, of the passenger nucleic acid portion as defined in claim 95 or 96.

138. A construct according to any one of claims 134 to 137, wherein the plurality of adjacent co-modified nucleotides are located in the nucleic acid linker portion as defined in claim 97(iii).

139. A construct according to any one of claims 128 to 138, wherein one or more modified nucleotides of the first nucleic acid portion of (a) do not have the co-modification present in the corresponding nucleotide of the third nucleic acid portion of (c) in the first duplex region; and / or (b) one or more modified nucleotides of the second nucleic acid portion do not have the co-modification present in the corresponding nucleotide of the fourth nucleic acid portion of (d) in the second duplex region; and / or one or more modified nucleotides of 1 to 8 additional nucleic acid portions, to the extent present as defined in claim 95 or 96, do not have the co-modification present in the corresponding nucleotide of the corresponding passenger nucleic acid portion in the respective duplex region.

140. A construct according to any one of claims 128 to 139, wherein one or more modified nucleotides of the first nucleic acid portion of (a) are shifted by at least one nucleotide relative to the co-modified nucleotides of the third nucleic acid portion of (c); and / or one or more modified nucleotides of the second nucleic acid portion of (b) are shifted by at least one nucleotide relative to the co-modified nucleotides of the fourth nucleic acid portion of (d); and / or one or more modified nucleotides of 1 to 8 additional nucleic acid portions (within the extent of presence defined in claim 95 or 96) are shifted by at least one nucleotide relative to the co-modified nucleotides of the passenger nucleic acid portion (within the extent of presence defined in claim 95 or 96).

141. A construct according to any one of claims 128 to 140, wherein the modification and / or modifications are each a sugar, phosphate or base modification.

142. A construct according to claim 141, wherein the modification is selected from nucleotides having a 2'-modified sugar; conformationally restricted nucleotide (CRN) sugars such as locked nucleic acid (LNA), (S)-constrained ethyl bicyclic nucleic acid and constrained ethyl (cEt), tricyclo-DNA; morpholino, unlocked nucleic acid (UNA), glycol nucleic acid (GNA), D-hexitol nucleic acid (HNA) and cyclohexene nucleic acid (CeNA).

143. The construct according to claim 142, wherein the 2'-modified sugar is selected from 2'-O-alkyl-modified sugars, 2'-O-methyl-modified sugars, 2'-O-methoxyethyl-modified sugars, 2'-O-allyl-modified sugars, 2'-C-allyl-modified sugars, 2'-deoxy-modified sugars such as 2'-deoxyribose, 2'-F-modified sugars, 2'-arabino-fluoride-modified sugars, 2'-O-benzyl-modified sugars, 2'-amino-modified sugars, and 2'-O-methyl-4-pyridyl-modified sugars.

144. The construct according to any one of claims 141 to 143, wherein the base modification is either a abasic nucleotide or a nucleotide containing a non-natural base.

145. The construct according to any one of claims 138 to 144, wherein at least one modification is a 2'-O-methyl modification in the ribose moiety.

146. The construct according to any one of claims 138 to 145, wherein at least one modification is a 2'-F modification of the ribose moiety.

147. The construct according to any one of claims 138 to 146, wherein (i) the first nucleic acid moiety of (a); and / or (ii) the second nucleic acid moiety of (b); and / or (iii) within the scope where present, the nucleotide at either position 2 or 14 downstream of the first nucleotide in the 5'-region of the 1 to 8 additional nucleic acid moieties defined in claim 95 or 96 does not contain a 2'-O-methyl modification in the ribose moiety.

148. The construct according to any one of claims 138 to 147, wherein (i) the third nucleic acid moiety of (c); and / or (ii) the fourth nucleic acid moiety of (d); and / or (iii) within the scope where present, the passenger nucleic acid moiety as defined in claim 95 or 96; which corresponds in position to (i) the first nucleic acid moiety of (a); and / or (ii) the second nucleic acid moiety of (b); and / or (iii) within the scope where present, any nucleotide at positions 11 to 13 of any nucleotide in the 5'-region of the 1 to 8 additional nucleic acid moieties defined in claim 95 or 96; does not contain a 2'-O-methyl modification in the ribose moiety.

149. The construct according to claim 147 or 148, wherein the nucleotide at either position 2 or 14 downstream of the first in (i) the first nucleic acid moiety of (a); and / or (ii) the second nucleic acid moiety of (b); and / or (iii) within the scope where present, the 1 to 8 additional nucleic acid moieties defined in claim 95 or 96; contains a 2'-F modification of the ribose moiety.

150. The construct according to any one of claims 147 to 149, wherein one, two, or all three nucleotides of (i) the third nucleic acid portion of (c); and / or (ii) the fourth nucleic acid portion of (d); and / or (iii), to the extent present, the passenger nucleic acid portion as defined in claim 95 or 96; are respectively located at positions corresponding to any nucleotide among nucleotides 11 to 13 downstream of the first nucleotide in the 5' region of (i) the first nucleic acid portion of (a); and / or (ii) the second nucleic acid portion of (b); and / or (iii), to the extent present, any of the 1 to 8 additional nucleic acid portions as defined in claim 95 or 96.

151. The construct according to any one of claims 146 to 150, wherein all remaining nucleotides contain a 2'-O-methyl modification or a 2'-F modification in the ribose moiety, preferably, except for the unmodified nucleotides according to claim 86.

152. The construct according to claim 151, wherein the remaining nucleotides contain a 2'-O-methyl modification in the ribose moiety.

153. The construct according to claim 151 or 152, wherein the one or more, preferably one, unmodified nucleotide represents any nucleotide of the nucleic acid linker portion as defined in claim 97(iii), preferably the nucleotide of the nucleic acid linker portion as defined in claim 97(iii), which is adjacent to (i) the third nucleic acid portion of (c); and / or (ii) the fourth nucleic acid portion of (d); and / or (iii), to the extent present, the passenger nucleic acid portion as defined in claim 95 or 96.

154. The construct of any of the preceding claims, wherein (a) the first nucleic acid portion is selected from Table 3a; (b) the second nucleic acid portion is selected from Table 3a; (c) the third nucleic acid portion is selected from Table 3b; and / or (d) the fourth nucleic acid portion is selected from Table 3b.

155. The construct according to claim 154, wherein the 3' terminal positions of the first and the third nucleic acid portions are replaced by unmodified nucleotides.

156. The construct according to any one of claims 82 to 155, which comprises at least one vinyl phosphonate modification, such as at least one vinyl phosphonate modification in the 5' region of (i) the first nucleic acid portion of (a); and / or (ii) the second nucleic acid portion of (b); and / or (iii) 1 to 8 additional nucleic acid portions as defined in claim 95 or 96 to the extent present.

157. The construct according to any one of claims 82 to 156, wherein the first nucleic acid portion of (a); and / or the second nucleic acid portion of (b); and / or the third nucleic acid portion of (c); and / or the fourth nucleic acid portion of (d); and / or 1 to 8 additional nucleic acid portions as defined in claim 95 or 96 to the extent present; and / or the passenger nucleic acid portion as defined in claim 95 or 96 to the extent present; is an inverted nucleotide and is linked to an adjacent nucleotide via the 3'-carbon of the nucleotide and the 3'-carbon of the adjacent nucleotide, and / or is an inverted nucleotide and is linked to an adjacent nucleotide via the 5'-carbon of the nucleotide and the 5'-carbon of the adjacent nucleotide.

158. The construct according to claim 157, wherein the inverted nucleotide is linked to the adjacent nucleotide via a phosphate group in a phosphodiester bond; or is linked to the adjacent nucleotide via a phosphorothioate group; or is linked to the adjacent nucleotide via a dithiophosphate group.

159. The construct according to any one of claims 82 to 158, which is blunt-ended.

160. The construct according to any one of claims 82 to 158, wherein (a) the first nucleic acid moiety; and / or (b) the second nucleic acid moiety; and / or (c) the third nucleic acid moiety; and / or (d) the fourth nucleic acid moiety; and / or to the extent present, 1 to 8 additional nucleic acid moieties as defined in claim 95 or 96; and / or to the extent present, a passenger nucleic acid moiety as defined in claim 95 or 96; has an overhang.

161. The construct according to any one of claims 82 to 160, wherein the target RNA is mRNA or another RNA molecule.

162. A composition comprising an oligomeric compound according to any one of claims 1 to 81 and / or a nucleic acid construct according to any one of claims 82 to 161, and a physiologically acceptable excipient.

163. A pharmaceutical composition comprising an oligomeric compound according to any one of claims 1 to 81 and / or a nucleic acid construct according to any one of claims 82 to 161.

164. The pharmaceutical composition of claim 163, further comprising a pharmaceutically acceptable excipient, diluent, antioxidant, and / or preservative.

165. The pharmaceutical composition of claim 163 or 164, wherein the oligomeric compound according to any one of claims 1 to 81 and / or the construct according to any one of claims 82 to 161 is the sole pharmaceutically active agent.

166. The pharmaceutical composition of claim 163 or 164, wherein the pharmaceutical composition further comprises one or more other pharmaceutically active agents.

167. The pharmaceutical composition of claim 166, wherein the additional pharmaceutically active agent is an agent that modulates the innate and / or adaptive immune system, such as an additional oligomeric compound directed against an immune system target other than complement component C5, preferably interleukin-6; an agent that reduces the expression or level of interleukin-6; or an agent such as an antibody directed against a complement component, said antibody preferably being eculizumab.

168. The pharmaceutical composition of claim 166 or 168, wherein the oligomeric compound and / or the nucleic acid construct; and the additional pharmaceutically active agent are administered simultaneously or in any order. An oligomeric compound according to any one of claims 1 to 81 and / or a nucleic acid construct according to any one of claims 82 to 161, for use in human or veterinary medicine or therapy. An oligomeric compound according to any one of claims 1 to 81 and / or a nucleic acid construct according to any one of claims 82 to 161, for use in a method of treating, ameliorating and / or preventing a disease or disorder.

171. The compound and / or construct according to claim 170, wherein the disease or disorder is a C5-related disease or disorder or a disease or disorder that requires reduction of C5 expression.

172. The compound and / or construct according to claim 171, wherein the disease or disorder is selected from autoimmune diseases, complement system dysfunction (including abnormal upregulation of complement components (such as C5)), age-related macular degeneration (AMD) (including dry AMD and geographic atrophy), paroxysmal nocturnal hemoglobinuria (PNH), generalized myasthenia gravis (gMG), lupus nephritis (LN), Alzheimer's disease, atherosclerosis, choroid plexus inflammation, atypical hemolytic uremic syndrome (aHUS), C3 glomerulopathy (C3G), Ig-mediated kidney diseases (such as IgA nephropathy and primary membranous nephropathy), asthma, rheumatism, rheumatoid arthritis, systemic lupus erythematosus (SLE), antineutrophil cytoplasmic antibody (ANCA) vasculitis, antiphospholipid antibody syndrome (APS), glomerulonephritis, bullous dermatomyositis pemphigoid, Shiga toxin Escherichia coli-related hemolytic uremic syndrome, amyotrophic lateral sclerosis (ALS), central nervous system (CNS) diseases, myasthenia gravis (MG), neuromyelitis optica (NMO), dense deposit disease, C3 neuropathy, cold agglutinin disease, humoral and vascular transplant rejection, graft dysfunction, myocardial infarction, asthma, rheumatoid arthritis (RA) transplant sensitization, antiphospholipid antibody syndrome; Lupus nephritis; ischemia-reperfusion injury; typical or atypical hemolytic uremic syndrome (tHUS); dense deposit disease (DDD); neuromyelitis optica (NO); multifocal motor neuropathy (MMN); multiple sclerosis (MS); macular degeneration (e.g., age-related macular degeneration (AMD)); hemolysis, elevated liver enzymes, and low platelets (HELLP) syndrome; thrombotic thrombocytopenic purpura (TTP); spontaneous fetal loss; pauci-immune vasculitis; epidermolysis bullosa; recurrent fetal loss; preeclampsia, traumatic brain injury, myasthenia gravis, cold agglutinin disease, dermatomyositis bullosa, Shiga toxin E.co / -associated hemolytic uremic syndrome, C3 glomerulopathy, antineutrophil cytoplasmic antibody-associated vasculitis, humoral and vascular transplant rejection, graft dysfunction, myocardial infarction, allograft, sepsis, coronary artery disease, dermatomyositis, Graves' disease, atherosclerosis, systemic inflammatory response sepsis, septic shock, spinal cord injury, glomerulonephritis, Hashimoto's thyroiditis, type I diabetes, pemphigus, autoimmune hemolytic anemia (AIHA), ITP, Goodpasture's syndrome, Degos' disease, antiphospholipid syndrome (APS), catastrophic APS (CAPS), cardiovascular disease, myocarditis, cerebrovascular disease, peripheral vascular disease, renal vascular disease, mesenteric / intestinal vascular disease, vasculitis, Henoch-Schönlein purpura nephritis, systemic lupus erythematosus-related vasculitis, vasculitis associated with rheumatoid arthritis, immune complex vasculitis, Takayasu's disease, dilated cardiomyopathy, diabetic angiopathy, Kawasaki disease (arteritis), venous gas embolism (VGE), restenosis after stent placement, rotational atherectomy, membranous nephropathy, Guillain-Barré syndrome, and percutaneous transluminal coronary angioplasty Age-related macular degeneration (AMD) and / or geographic atrophy (GA); uveitis and / or panuveitis; cold agglutinin disease, membranoproliferative glomerulonephritis (MPGN), Guillain-Barré syndrome, Shiga toxin-producing Escherichia coli hemolytic uremic syndrome (STEC-HUS), organ transplantation-related autoimmune diseases, and sepsis.

173. A compound for use according to claim 171 or 172, wherein the disease or disorder is selected from paroxysmal nocturnal hemoglobinuria (PNH), Alzheimer's disease, atherosclerosis, choroid plexus inflammation, generalized myasthenia gravis (gMG), amyotrophic lateral sclerosis (ALS), lupus nephritis (LN), central nervous system (CNS) diseases; age-related macular degeneration (AMD) and / or geographic atrophy (GA); uveitis and / or panuveitis; cold agglutinin disease, membranoproliferative glomerulonephritis (MPGN), Guillain-Barré syndrome, Shiga toxin-producing Escherichia coli hemolytic uremic syndrome (STEC-HUS), and organ transplantation-related autoimmune diseases.

174. A nucleic acid construct and / or oligomeric compound for use according to any one of claims 169 to 173, wherein the dose of the nucleic acid construct and / or oligomeric compound is administered at a dose of about 0.05 mg / kg to about 50.0 mg / kg, optionally 0.05 mg / kg to about 30.0 mg / kg or 10.0 mg / kg to about 50.0 mg / kg, based on the body weight of a human subject.

175. A method of treating a disease or disorder, comprising administering to an individual in need thereof an oligomeric compound according to any one of claims 1 to 81 and / or a nucleic acid construct according to any one of claims 82 to 161.

176. The method according to claim 174, wherein the oligomeric compound and / or nucleic acid construct is administered subcutaneously or intravenously to the individual.

177. Use of an oligomeric compound according to any one of claims 1 to 81 or a nucleic acid construct according to any one of claims 82 to 161 as a gene function analysis tool for research.

178. Use of an oligomeric compound according to any one of claims 1 to 81 and / or a nucleic acid construct according to any one of claims 82 to 161 in the manufacture of a medicament for treating a disease or disorder.

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