COMPLEMENT COMPONENT C5 iRNA COMPOSITIONS AND METHODS OF USE THEREOF FOR TREATING PAROXYSMAL NOCTURNAL HEMOGLOBINURIA (PNH)

Abstract

To provide methods for treating subjects having paroxysmal nocturnal hemoglobinuria.SOLUTION: A method for treating a subject having paroxysmal nocturnal hemoglobinuria (PNH) comprises administering to an eculizumab-naive subject a 200-400 mg fixed dose of a double stranded ribonucleic acid (dsRNA) agent for inhibiting expression of complement component C5 once every week; and administering to the subject a dose of about 300 mg to about 900 mg of eculizumab, or an antigen-binding fragment thereof, thereby treating the subject. The dsRNA agent comprises a sense strand and an antisense strand. The sense strand comprises 5'-asasGfcAfaGfaUfAfUfuUfuuAfuAfaua-3'. The antisense strand comprises 5'-usAfsUfuAfuaAfaAfauaUfcUfuGfcuususudTdT-3'.SELECTED DRAWING: None

Description

Technical Field

[0001] Related Applications This application claims the benefit of priority of U.S. Provisional Patent Application No. 62 / 348,564, filed on June 10, 2016, and U.S. Provisional Patent Application No. 62 / 429,448, filed on December 2, 2016. The entire contents of each of the above provisional patent applications are incorporated herein by reference.

[0002] Sequence Listing This application includes a sequence listing submitted electronically in ASCII format, which is incorporated herein by reference in its entirety. The name of the ASCII copy created on May 31, 2017 is 121301-06420_SL.txt, and the size is 956,487 bytes.

Background Art

[0003] Complement was first discovered in the 1890s (Non-Patent Document 1) when it was found to assist or "complement" the killing of bacteria by heat-stable antibodies present in normal serum. The complement system consists of more than 30 proteins that exist as soluble proteins in the blood or as membrane-bound proteins. Activation of complement results in a sequential cascade of enzymatic reactions known as the complement activation pathway, leading to the formation of the potent anaphylatoxins C3a and C5a, which trigger many physiological reactions ranging from chemotaxis to apoptosis. Initially, complement was thought to play a major role in innate immunity where a strong and rapid response is made against invading pathogens. However, more recently, complement has been shown to play an important role in adaptive immunity involving T cells and B cells that help eliminate pathogens in maintaining immune memory against reinvasion of pathogens (Non-Patent Document 2; Non-Patent Document 3), and is increasingly being shown to be involved in human pathological conditions (Non-Patent Document 4; Non-Patent Document 5).

[0004] Complement activation is known to occur through three distinct pathways involving proteins that mainly exist as inactive zymogens and are subsequently sequentially cleaved and activated: the alternative pathway, the classical pathway, and the lectin pathway (Figure 1). All pathways of complement activation result in the cleavage of C5 molecule to generate anaphylatoxin C5a and C5b which later forms the terminal complement complex (C5b-9). C5a exerts significant pro-inflammatory activity through interaction with the classical G-protein coupled receptor C5aR (CD88) and the non-G-protein coupled receptor C5L2 (GPR77), which are expressed on various immune and non-immune cells. C5b-9 causes cell lysis through the formation of the membrane attack complex (MAC), and sub-lytic MAC and soluble C5b-9 also have many non-cytolytic immune functions. These two complement effectors, C5a and C5b-9, generated from the cleavage of C5, are major components of the complement system that are responsible for the propagation and / or initiation of lesions in various diseases including paroxysmal nocturnal hemoglobinuria, rheumatoid arthritis, ischemia-reperfusion injury and neurodegenerative diseases.

[0005] To date, the anti-C5 antibody, eculizumab (Soliris®), which is the only therapy targeting the C5-C5a axis, is available for the treatment of diseases associated with complement component C5. Eculizumab has been shown to be effective in the treatment of paroxysmal nocturnal hemoglobinuria (PNH) and atypical hemolytic uremic syndrome (aHUS), and is currently being evaluated in clinical trials for additional diseases associated with complement component C5. However, the eculizumab therapy requires a high dose injection once a week and subsequent bi-weekly maintenance injections at an annual cost of approximately $400,000. Furthermore, there are large individual differences in the pharmacodynamics and clearance of eculizumab, and a significant number of patients treated with eculizumab still require transfusion while undergoing treatment due to hemolytic attacks or potential hemolysis (Non-Patent Document 6; Non-Patent Document 7; Non-Patent Document 8). Therefore, there is a need in the art for alternative and combination therapies that provide a fairly consistent level of efficacy and minimal hemolytic attacks or potential hemolysis for subjects suffering from diseases associated with complement component C5.

Prior Art Documents

Non-Patent Documents

[0006]

Non-Patent Document 1

Non-Patent Document 2

Non-Patent Document 3

Non-Patent Document 4

Non-Patent Document 5

Non-Patent Document 6

Non-Patent Document 7

Non-Patent Document 8

Summary of the Invention

Means for Solving the Problems

[0007] The present invention provides an iRNA composition that results in cleavage via an RNA-induced silencing complex (RISC) of an RNA transcript of a C5 gene to inhibit the expression of the C5 gene, and a method and combination therapy for treating a subject suffering from a disorder, such as paroxysmal nocturnal hemoglobinuria (PNH), atypical hemolytic uremic syndrome (aHUS), neuromyelitis optica (NMO), and myasthenia gravis, that may benefit from inhibiting or reducing the expression of the C5 gene using an anti-C5 antibody, such as eculizumab.

[0008] The data presented herein demonstrate that the iRNA agents and compositions of the present invention are effective in treating PNH in eculizumab-naïve subjects when administered as monotherapy at doses of 200 mg or 400 mg. The data presented herein also demonstrate that the iRNA agents and compositions of the present invention can be effectively used as part of combination therapy with eculizumab for treating subjects suffering from PNH. For example, in the current AD-62643 pharmacology situation, administering the iRNA agents and compositions of the present invention in combination with eculizumab allows for a reduction in the dose of eculizumab while maintaining C5 knockdown, enabling inhibition of complement activity and a decrease in LDH levels in subjects suffering from PNH. The data also demonstrate that the iRNA agents and compositions of the present invention are suitable for treating subjects suffering from PNH who are non-responsive to treatment with eculizumab alone (e.g., subjects with hemolytic attacks, i.e., subjects being treated with eculizumab who experienced symptoms of intravascular hemolysis 1 - 2 days prior to the next eculizumab infusion).

[0009] Accordingly, in one aspect, the present invention provides a method for treating, e.g., treating for a long period of time, a subject suffering from paroxysmal nocturnal hemoglobinuria (PNH). The method comprises administering, once a week, a fixed dose of 200 - 400 mg of a double-stranded ribonucleic acid (dsRNA) agent that inhibits the expression of complement component C5 to a subject with PNH who has not been administered eculizumab (i.e., a subject suffering from PNH who has not been administered eculizumab); and administering to the subject a dose of eculizumab, or an antigen-binding fragment thereof, of about 300 mg to about 900 mg, thereby treating the subject, wherein the dsRNA agent comprises a sense strand and an antisense strand, wherein the sense strand comprises 5’-asasGfcAfaGfaUfAfUfuUfuuAfuAfaua-3’ (SEQ ID NO: 2876), and the antisense strand comprises 5’-usAfsUfuAfuaAfaAfauaUfcUfuGfcuususudTdT-3’ (SEQ ID NO: 2889), wherein a, g, c, and u are 2’-O-methyl (2’-OMe) A, G, C, and U, respectively; Af, Gf, Cf, and Uf are 2’-fluoro A, G, C, and U, respectively; dT is a deoxy-thymine nucleotide; and s is a phosphorothioate bond.

[0010] In another aspect, the present invention provides a method for treating, e.g., treating over a long period of time, a subject suffering from paroxysmal nocturnal hemoglobinuria (PNH). The method comprises administering to a subject with PNH who has not been administered eculizumab a double-stranded ribonucleic acid (dsRNA) agent at a fixed dose of 200-400 mg once a month for inhibiting the expression of complement component C5; and administering to the subject a dose of eculizumab, or an antigen-binding fragment thereof, of about 300 mg to about 900 mg, thereby treating the subject, wherein the dsRNA agent comprises a sense strand and an antisense strand, wherein the sense strand comprises 5’-asasGfcAfaGfaUfAfUfuUfuuAfuAfaua-3’ (SEQ ID NO: 2876), and the antisense strand comprises 5’-usAfsUfuAfuaAfaAfauaUfcUfuGfcuususudTdT-3’ (SEQ ID NO: 2889), wherein a, g, c, and u are 2’-O-methyl (2’-OMe) A, G, C, and U, respectively; Af, Gf, Cf, and Uf are 2’-fluoro A, G, C, and U, respectively; dT is a deoxy-thymine nucleotide; and s is a phosphorothioate bond.

[0011] In one aspect, the present invention provides a method for treating, e.g., treating for a long period of time, a subject suffering from paroxysmal nocturnal hemoglobinuria (PNH). The method comprises administering to a subject with PNH who has not received eculizumab a double-stranded ribonucleic acid (dsRNA) agent at a fixed dose of 200 mg once a week for 10 to 15 weeks to inhibit the expression of complement component C5, followed by administering a dsRNA agent at a fixed dose of 400 mg once a week; and administering to the subject an eculizumab at a dose of about 300 mg to about 900 mg, or an antigen-binding fragment thereof, thereby treating the subject, wherein the dsRNA agent comprises a sense strand and an antisense strand, wherein the sense strand comprises 5'-asasGfcAfaGfaUfAfUfuUfuuAfuAfaua-3' (SEQ ID NO: 2876), and the antisense strand comprises 5'-usAfsUfuAfuaAfaAfauaUfcUfuGfcuususudTdT-3' (SEQ ID NO: 2889), wherein a, g, c, and u are 2'-O-methyl (2'-OMe) A, G, C, and U, respectively; Af, Gf, Cf, and Uf are 2'-fluoro A, G, C, and U, respectively; dT is deoxy-thymidine nucleotide; and s is a phosphorothioate bond. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 200 mg once a week for 13 weeks, followed by a dsRNA agent at a fixed dose of 400 mg once a week.

[0012] In one aspect, the present invention provides a method for treating, e.g., treating over a long period of time, a subject suffering from paroxysmal nocturnal hemoglobinuria (PNH). The method comprises administering to a subject with PNH who has not been administered eculizumab a double-stranded ribonucleic acid (dsRNA) agent at a fixed dose of 200 mg once a week for 10 to 15 weeks to inhibit the expression of complement component C5, followed by administering a dsRNA agent at a fixed dose of 400 mg once a month; and administering to the subject a dose of eculizumab or an antigen-binding fragment thereof of about 300 mg to about 900 mg, thereby treating the subject, wherein the dsRNA agent comprises a sense strand and an antisense strand, wherein the sense strand comprises 5’-asasGfcAfaGfaUfAfUfuUfuuAfuAfaua-3’ (SEQ ID NO: 2876), and the antisense strand comprises 5’-usAfsUfuAfuaAfaAfauaUfcUfuGfcuususudTdT-3’ (SEQ ID NO: 2889), wherein a, g, c, and u are 2’-O-methyl (2’-OMe) A, G, C, and U, respectively; Af, Gf, Cf, and Uf are 2’-fluoro A, G, C, and U, respectively; dT is deoxy-thymidine nucleotide; and s is a phosphorothioate bond. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 200 mg once a week for 13 weeks, followed by a dsRNA agent at a fixed dose of 400 mg once a month.

[0013] In another aspect, the present invention provides a method for treating, e.g., treating over a long period of time, a subject suffering from paroxysmal nocturnal hemoglobinuria (PNH). The method comprises administering to a subject with PNH who has not received eculizumab a double-stranded ribonucleic acid (dsRNA) agent at a fixed dose of 200 mg once a month for 2 to 4 months to inhibit the expression of complement component C5; and administering to the subject a dose of eculizumab, or an antigen-binding fragment thereof, of about 300 mg to about 900 mg, thereby treating the subject, wherein the dsRNA agent comprises a sense strand and an antisense strand, wherein the sense strand comprises 5’-asasGfcAfaGfaUfAfUfuUfuuAfuAfaua-3’ (SEQ ID NO: 2876), and the antisense strand comprises 5’-usAfsUfuAfuaAfaAfauaUfcUfuGfcuususudTdT-3’ (SEQ ID NO: 2889), wherein a, g, c, and u are 2’-O-methyl (2’-OMe) A, G, C, and U, respectively; Af, Gf, Cf, and Uf are 2’-fluoro A, G, C, and U, respectively; dT is deoxy-thymine nucleotide; and s is a phosphorothioate linkage.

[0014] In one aspect, the present invention provides a method for treating, e.g., treating over a long period of time, a subject suffering from paroxysmal nocturnal hemoglobinuria (PNH). The method comprises administering to a naïve subject suffering from PNH a double-stranded ribonucleic acid (dsRNA) agent at a fixed dose of 200 mg once a week for inhibiting the expression of complement component C5; and administering to the subject eculizumab or an antigen-binding fragment thereof at a dose of about 300 mg to about 900 mg once every four weeks, thereby treating the subject, wherein the dsRNA agent comprises a sense strand and an antisense strand, wherein the sense strand comprises 5’-asasGfcAfaGfaUfAfUfuUfuuAfuAfaua-3’ (SEQ ID NO: 2876), and the antisense strand comprises 5’-usAfsUfuAfuaAfaAfauaUfcUfuGfcuususudTdT-3’ (SEQ ID NO: 2889), wherein a, g, c, and u are 2’-O-methyl (2’-OMe) A, G, C, and U, respectively; Af, Gf, Cf, and Uf are 2’-fluoro A, G, C, and U, respectively; dT is deoxy-thymidine nucleotide; and s is a phosphorothioate bond. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 200 mg once a week for 8 weeks, e.g., prior to the administration of eculizumab. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 200 mg once a week for 8 weeks, e.g., prior to the administration of eculizumab, and then once a month thereafter. In another embodiment, the dsRNA agent is administered to the subject at a fixed dose of 200 mg once a week for 8 weeks, e.g., prior to the administration of eculizumab, and then once every three months thereafter. In another embodiment, the dsRNA agent is administered to the subject at a fixed dose of 200 mg once a week for 8 weeks, e.g., prior to the administration of eculizumab, and then once every six months thereafter. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 200 mg once a week for a long period of time.

[0015] In one aspect, the present invention provides a method for treating, e.g., treating over a long period of time, a subject suffering from paroxysmal nocturnal hemoglobinuria (PNH). The method comprises administering to a subject not receiving eculizumab and suffering from PNH a double-stranded ribonucleic acid (dsRNA) agent at a fixed dose of 200 mg once a week for inhibiting the expression of complement component C5; and administering to the subject eculizumab or an antigen-binding fragment thereof at a dose of about 300 mg once every 4 weeks, thereby treating the subject, wherein the dsRNA agent comprises a sense strand and an antisense strand, wherein the sense strand comprises 5'-asasGfcAfaGfaUfAfUfuUfuuAfuAfaua-3' (SEQ ID NO: 2876), the antisense strand comprises 5'-usAfsUfuAfuaAfaAfauaUfcUfuGfcuususudTdT-3' (SEQ ID NO: 2889), wherein a, g, c, and u are 2'-O-methyl (2'-OMe) A, G, C, and U, respectively; Af, Gf, Cf, and Uf are 2'-fluoro A, G, C, and U, respectively; dT is deoxy-thymine nucleotide; and s is a phosphorothioate bond. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 200 mg once a week for 8 weeks, e.g., before the administration of eculizumab. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 200 mg once a week for 8 weeks, e.g., before the administration of eculizumab, and then once a month thereafter. In another embodiment, the dsRNA agent is administered to the subject at a fixed dose of 200 mg once a week for 8 weeks, e.g., before the administration of eculizumab, and then once every 3 months thereafter. In another embodiment, the dsRNA agent is administered to the subject at a fixed dose of 200 mg once a week for 8 weeks, e.g., before the administration of eculizumab, and then once every 6 months thereafter. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 200 mg once a week for a long period of time.

[0016] In one aspect, the present invention provides a method for treating, e.g., treating over a long period of time, a subject suffering from paroxysmal nocturnal hemoglobinuria (PNH). The method comprises administering to a subject not previously administered eculizumab suffering from PNH, once a week, a fixed dose of 200 mg of a double-stranded ribonucleic acid (dsRNA) agent for inhibiting the expression of complement component C5; and administering to the subject, once every 4 weeks, a dose of about 600 mg of eculizumab, or an antigen-binding fragment thereof, thereby treating the subject, wherein the dsRNA agent comprises a sense strand and an antisense strand, wherein the sense strand comprises 5’-asasGfcAfaGfaUfAfUfuUfuuAfuAfaua-3’ (SEQ ID NO: 2876), and the antisense strand comprises 5’-usAfsUfuAfuaAfaAfauaUfcUfuGfcuususudTdT-3’ (SEQ ID NO: 2889), wherein a, g, c, and u are 2’-O-methyl (2’-OMe) A, G, C, and U, respectively; Af, Gf, Cf, and Uf are 2’-fluoro A, G, C, and U, respectively; dT is a deoxy-thymine nucleotide; and s is a phosphorothioate bond. In one embodiment, the dsRNA agent is administered to the subject once a week, at a fixed dose of 200 mg, for 8 weeks, e.g., prior to the administration of eculizumab. In one embodiment, the dsRNA agent is administered to the subject once a week, at a fixed dose of 200 mg, for 8 weeks, e.g., prior to the administration of eculizumab, and then once a month thereafter. In another embodiment, the dsRNA agent is administered to the subject once a week, at a fixed dose of 200 mg, for 8 weeks, e.g., prior to the administration of eculizumab, and then once every 3 months thereafter. In another embodiment, the dsRNA agent is administered to the subject once a week, at a fixed dose of 200 mg, for 8 weeks, e.g., prior to the administration of eculizumab, and then once every 6 months thereafter. In one embodiment, the dsRNA agent is administered to the subject over a long period of time, once a week, at a fixed dose of 200 mg.

[0017] In one aspect, the present invention provides a method for treating, e.g., treating over a long period of time, a subject suffering from paroxysmal nocturnal hemoglobinuria (PNH). The method comprises administering to a subject with PNH who has not received eculizumab a double-stranded ribonucleic acid (dsRNA) agent at a fixed dose of 200 mg once a week for inhibiting the expression of complement component C5; and administering to the subject eculizumab or an antigen-binding fragment thereof at a dose of about 900 mg once every 4 weeks, thereby treating the subject, wherein the dsRNA agent comprises a sense strand and an antisense strand, wherein the sense strand comprises 5'-asasGfcAfaGfaUfAfUfuUfuuAfuAfaua-3' (SEQ ID NO: 2876), and the antisense strand comprises 5'-usAfsUfuAfuaAfaAfauaUfcUfuGfcuususudTdT-3' (SEQ ID NO: 2889), wherein a, g, c, and u are 2'-O-methyl (2'-OMe) A, G, C, and U, respectively; Af, Gf, Cf, and Uf are 2'-fluoro A, G, C, and U, respectively; dT is deoxy-thymine nucleotide; and s is a phosphorothioate bond. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 200 mg once a week for 8 weeks, e.g., before the administration of eculizumab. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 200 mg once a week for 8 weeks, e.g., before the administration of eculizumab, and then once a month thereafter. In another embodiment, the dsRNA agent is administered to the subject at a fixed dose of 200 mg once a week for 8 weeks, e.g., before the administration of eculizumab, and then once every 3 months thereafter. In another embodiment, the dsRNA agent is administered to the subject at a fixed dose of 200 mg once a week for 8 weeks, e.g., before the administration of eculizumab, and then once every 6 months thereafter. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 200 mg once a week for a long period of time.

[0018] In another aspect, the present invention provides a method for treating, e.g., treating over a long period of time, a subject suffering from paroxysmal nocturnal hemoglobinuria (PNH). The method comprises administering to a subject with PNH who has not received eculizumab a double-stranded ribonucleic acid (dsRNA) agent at a fixed dose of 200 mg once a month to inhibit the expression of complement component C5; and administering to the subject eculizumab or an antigen-binding fragment thereof at a dose of about 300 mg to about 900 mg once every four weeks, thereby treating the subject, wherein the dsRNA agent comprises a sense strand and an antisense strand, wherein the sense strand comprises 5'-asasGfcAfaGfaUfAfUfuUfuuAfuAfaua-3' (SEQ ID NO: 2876), and the antisense strand comprises 5'-usAfsUfuAfuaAfaAfauaUfcUfuGfcuususudTdT-3' (SEQ ID NO: 2889), wherein a, g, c, and u are 2'-O-methyl (2'-OMe) A, G, C, and U, respectively; Af, Gf, Cf, and Uf are 2'-fluoro A, G, C, and U, respectively; dT is deoxy-thymine nucleotide; and s is a phosphorothioate bond. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 200 mg once a month for two months, e.g., prior to the administration of eculizumab. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 200 mg once a month for two months, e.g., prior to the administration of eculizumab, and then once every three months. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 200 mg once a month for two months, e.g., prior to the administration of eculizumab, and then once every six months. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 200 mg once a month for a long period of time.

[0019] In another aspect, the present invention provides a method for treating, e.g., treating for a long period of time, a subject suffering from paroxysmal nocturnal hemoglobinuria (PNH). The method comprises administering to a subject with PNH who has not been administered eculizumab a double-stranded ribonucleic acid (dsRNA) agent at a fixed dose of 200 mg once a month for inhibiting the expression of complement component C5; and administering to the subject eculizumab or an antigen-binding fragment thereof at a dose of about 300 mg once every four weeks, thereby treating the subject, wherein the dsRNA agent comprises a sense strand and an antisense strand, wherein the sense strand comprises 5’-asasGfcAfaGfaUfAfUfuUfuuAfuAfaua-3’ (SEQ ID NO: 2876), and the antisense strand comprises 5’-usAfsUfuAfuaAfaAfauaUfcUfuGfcuususudTdT-3’ (SEQ ID NO: 2889), wherein a, g, c, and u are 2’-O-methyl (2’-OMe) A, G, C, and U, respectively; Af, Gf, Cf, and Uf are 2’-fluoro A, G, C, and U, respectively; dT is deoxy-thymine nucleotide; and s is a phosphorothioate bond. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 200 mg once a month for two months, e.g., prior to the administration of eculizumab. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 200 mg once a month for two months, e.g., prior to the administration of eculizumab, and then once every three months. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 200 mg once a month for two months, e.g., prior to the administration of eculizumab, and then once every six months. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 200 mg once a month for a long period of time.

[0020] In another aspect, the present invention provides a method for treating, e.g., treating over a long period of time, a subject suffering from paroxysmal nocturnal hemoglobinuria (PNH). The method comprises administering to a subject not receiving eculizumab and suffering from PNH a double-stranded ribonucleic acid (dsRNA) agent at a fixed dose of 200 mg once a month for inhibiting the expression of complement component C5; and administering to the subject eculizumab or an antigen-binding fragment thereof at a dose of about 600 mg once every four weeks, thereby treating the subject, wherein the dsRNA agent comprises a sense strand and an antisense strand, wherein the sense strand comprises 5’-asasGfcAfaGfaUfAfUfuUfuuAfuAfaua-3’ (SEQ ID NO: 2876), and the antisense strand comprises 5’-usAfsUfuAfuaAfaAfauaUfcUfuGfcuususudTdT-3’ (SEQ ID NO: 2889), wherein a, g, c, and u are 2’-O-methyl (2’-OMe) A, G, C, and U, respectively; Af, Gf, Cf, and Uf are 2’-fluoro A, G, C, and U, respectively; dT is deoxy-thymidine nucleotide; and s is a phosphorothioate linkage. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 200 mg of the dsRNA agent once a month for two months, e.g., prior to the administration of eculizumab. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 200 mg of the dsRNA agent once a month for two months, e.g., prior to the administration of eculizumab, and then once every three months. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 200 mg of the dsRNA agent once a month for two months, e.g., prior to the administration of eculizumab, and then once every six months. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 200 mg of the dsRNA agent once a month for a long period of time.

[0021] In another aspect, the present invention provides a method for treating, e.g., treating over a long period of time, a subject suffering from paroxysmal nocturnal hemoglobinuria (PNH). The method comprises administering to a subject with PNH who has not received eculizumab a double-stranded ribonucleic acid (dsRNA) agent at a fixed dose of 200 mg once a month for inhibiting the expression of complement component C5; and administering to the subject eculizumab or an antigen-binding fragment thereof at a dose of about 900 mg once every four weeks, thereby treating the subject, wherein the dsRNA agent comprises a sense strand and an antisense strand, wherein the sense strand comprises 5’-asasGfcAfaGfaUfAfUfuUfuuAfuAfaua-3’ (SEQ ID NO: 2876), and the antisense strand comprises 5’-usAfsUfuAfuaAfaAfauaUfcUfuGfcuususudTdT-3’ (SEQ ID NO: 2889), Here, a, g, c, and u are 2'-O-methyl (2'-OMe) A, G, C, and U, respectively; Af, Gf, Cf, and Uf are 2'-fluoro A, G, C, and U, respectively; dT is deoxy-thymidine nucleotide; and s is a phosphorothioate bond. In one embodiment, the dsRNA agent is administered to a subject at a fixed dose of 200 mg once a month for 2 months, for example, before the administration of eculizumab. In one embodiment, the dsRNA agent is administered to a subject at a fixed dose of 200 mg once a month for 2 months, for example, before the administration of eculizumab, and then once every 3 months thereafter. In one embodiment, the dsRNA agent is administered to a subject at a fixed dose of 200 mg once a month for 2 months, for example, before the administration of eculizumab, and then once every 6 months thereafter. In one embodiment, the dsRNA agent is administered to a subject at a fixed dose of 200 mg once a month for a long period. In one aspect, the present invention provides a method for treating, for example, treating for a long period, a subject suffering from paroxysmal nocturnal hemoglobinuria (PNH). The method comprises administering to a PNH subject not administered with eculizumab a double-stranded ribonucleic acid (dsRNA) agent at a fixed dose of 400 mg once a week for inhibiting the expression of complement component C5; and administering to the subject eculizumab or an antigen-binding fragment thereof at a dose of about 300 mg to about 900 mg once every 4 weeks, thereby treating the subject, wherein the dsRNA agent comprises a sense strand and an antisense strand, wherein the sense strand comprises 5'-asasGfcAfaGfaUfAfUfuUfuuAfuAfaua-3' (SEQ ID NO: 2876), and the antisense strand comprises 5'-usAfsUfuAfuaAfaAfauaUfcUfuGfcuususudTdT-3' (SEQ ID NO: 2889), here, a, g, c, and u are 2'-O-methyl (2'-OMe) A, G, C, and U, respectively; Af, Gf, Cf, and Uf are 2'-fluoro A, G, C, and U, respectively; dT is deoxy-thymidine nucleotide; and s is a phosphorothioate bond.In one embodiment, the dsRNA agent is administered to the subject, for example, at a fixed dose of 400 mg of the dsRNA agent once a week for 8 weeks, prior to the administration of eculizumab. In one embodiment, the dsRNA agent is administered to the subject, for example, at a fixed dose of 400 mg of the dsRNA agent once a week for 8 weeks, and then once a month, prior to the administration of eculizumab. In one embodiment, the dsRNA agent is administered to the subject, for example, at a fixed dose of 400 mg of the dsRNA agent once a week for 8 weeks, and then once every three months, prior to the administration of eculizumab. In one embodiment, the dsRNA agent is administered to the subject, for example, at a fixed dose of 400 mg of the dsRNA agent once a week for 8 weeks, and then once every six months, prior to the administration of eculizumab. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 400 mg of the dsRNA agent once a week for a long period of time.

[0022] In one aspect, the present invention provides a method for treating, e.g., treating over a long period of time, a subject suffering from paroxysmal nocturnal hemoglobinuria (PNH). The method comprises administering to a naïve subject suffering from PNH a double-stranded ribonucleic acid (dsRNA) agent at a fixed dose of 400 mg once a week for inhibiting the expression of complement component C5; and administering to the subject eculizumab or an antigen-binding fragment thereof at a dose of about 300 mg once every 4 weeks, thereby treating the subject, wherein the dsRNA agent comprises a sense strand and an antisense strand, wherein the sense strand comprises 5’-asasGfcAfaGfaUfAfUfuUfuuAfuAfaua-3’ (SEQ ID NO: 2876), and the antisense strand comprises 5’-usAfsUfuAfuaAfaAfauaUfcUfuGfcuususudTdT-3’ (SEQ ID NO: 2889), wherein a, g, c, and u are 2’-O-methyl (2’-OMe) A, G, C, and U, respectively; Af, Gf, Cf, and Uf are 2’-fluoro A, G, C, and U, respectively; dT is a deoxy-thymine nucleotide; and s is a phosphorothioate bond. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 400 mg once a week for 8 weeks prior to, e.g., the administration of eculizumab. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 400 mg once a week for 8 weeks prior to, e.g., the administration of eculizumab and then once a month thereafter. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 400 mg once a week for 8 weeks prior to, e.g., the administration of eculizumab and then once every 3 months thereafter. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 400 mg once a week for 8 weeks prior to, e.g., the administration of eculizumab and then once every 6 months thereafter. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 400 mg once a week for a long period of time.

[0023] In one aspect, the present invention provides a method for treating, e.g., treating over a long period of time, a subject suffering from paroxysmal nocturnal hemoglobinuria (PNH). The method comprises administering to a subject with PNH who has not received eculizumab a double-stranded ribonucleic acid (dsRNA) agent at a fixed dose of 400 mg once a week for inhibiting the expression of complement component C5; and administering to the subject eculizumab or an antigen-binding fragment thereof at a dose of about 600 mg once every 4 weeks, thereby treating the subject, wherein the dsRNA agent comprises a sense strand and an antisense strand, wherein the sense strand comprises 5’-asasGfcAfaGfaUfAfUfuUfuuAfuAfaua-3’ (SEQ ID NO: 2876), and the antisense strand comprises 5’-usAfsUfuAfuaAfaAfauaUfcUfuGfcuususudTdT-3’ (SEQ ID NO: 2889), wherein a, g, c, and u are 2’-O-methyl (2’-OMe) A, G, C, and U, respectively; Af, Gf, Cf, and Uf are 2’-fluoro A, G, C, and U, respectively; dT is a deoxy-thymine nucleotide; and s is a phosphorothioate bond. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 400 mg once a week for 8 weeks, e.g., prior to the administration of eculizumab. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 400 mg once a week for 8 weeks, e.g., prior to the administration of eculizumab, and then once a month thereafter. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 400 mg once a week for 8 weeks, e.g., prior to the administration of eculizumab, and then once every 3 months thereafter. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 400 mg once a week for 8 weeks, e.g., prior to the administration of eculizumab, and then once every 6 months thereafter. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 400 mg once a week for a long period of time.

[0024] In one aspect, the present invention provides a method for treating, e.g., treating over a long period of time, a subject suffering from paroxysmal nocturnal hemoglobinuria (PNH). The method comprises administering to a subject not previously administered eculizumab suffering from PNH a double-stranded ribonucleic acid (dsRNA) agent at a fixed dose of 400 mg once a week for inhibiting the expression of complement component C5; and administering to the subject eculizumab or an antigen-binding fragment thereof at a dose of about 900 mg once every four weeks, thereby treating the subject, wherein the dsRNA agent comprises a sense strand and an antisense strand, wherein the sense strand comprises 5'-asasGfcAfaGfaUfAfUfuUfuuAfuAfaua-3' (SEQ ID NO: 2876), and the antisense strand comprises 5'-usAfsUfuAfuaAfaAfauaUfcUfuGfcuususudTdT-3' (SEQ ID NO: 2889), wherein a, g, c, and u are 2'-O-methyl (2'-OMe) A, G, C, and U, respectively; Af, Gf, Cf, and Uf are 2'-fluoro A, G, C, and U, respectively; dT is deoxy-thymidine nucleotide; and s is a phosphorothioate bond. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 400 mg once a week for 8 weeks, e.g., prior to the administration of eculizumab. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 400 mg once a week for 8 weeks, e.g., prior to the administration of eculizumab, and then once a month thereafter. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 400 mg once a week for 8 weeks, e.g., prior to the administration of eculizumab, and then once every three months thereafter. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 400 mg once a week for 8 weeks, e.g., prior to the administration of eculizumab, and then once every six months thereafter. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 400 mg once a week for a long period of time.

[0025] In another aspect, the present invention provides a method for treating, e.g., treating over a long period of time, a subject suffering from paroxysmal nocturnal hemoglobinuria (PNH). The method comprises administering to a subject with PNH who has not been administered eculizumab a double-stranded ribonucleic acid (dsRNA) agent at a fixed dose of 400 mg once a month to inhibit the expression of complement component C5; and administering to the subject eculizumab or an antigen-binding fragment thereof at a dose of about 300 mg to about 900 mg once every four weeks, thereby treating the subject, wherein the dsRNA agent comprises a sense strand and an antisense strand, wherein the sense strand comprises 5’-asasGfcAfaGfaUfAfUfuUfuuAfuAfaua-3’ (SEQ ID NO: 2876), and the antisense strand comprises 5’-usAfsUfuAfuaAfaAfauaUfcUfuGfcuususudTdT-3’ (SEQ ID NO: 2889), wherein a, g, c, and u are 2’-O-methyl (2’-OMe) A, G, C, and U, respectively; Af, Gf, Cf, and Uf are 2’-fluoro A, G, C, and U, respectively; dT is deoxy-thymidine nucleotide; and s is a phosphorothioate bond. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 400 mg once a month for two months, e.g., prior to the administration of eculizumab. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 400 mg once a month for two months, e.g., prior to the administration of eculizumab, and then once every three months. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 400 mg once a month for two months, e.g., prior to the administration of eculizumab, and then once every six months. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 400 mg once a month for a long period of time.

[0026] In another aspect, the present invention provides a method for treating, e.g., treating over a long period of time, a subject suffering from paroxysmal nocturnal hemoglobinuria (PNH). The method comprises administering to a subject with PNH who has not been administered eculizumab a double-stranded ribonucleic acid (dsRNA) agent at a fixed dose of 400 mg once a month for inhibiting the expression of complement component C5; and administering to the subject a dose of about 300 mg of eculizumab, or an antigen-binding fragment thereof, thereby treating the subject, wherein the dsRNA agent comprises a sense strand and an antisense strand, wherein the sense strand comprises 5’-asasGfcAfaGfaUfAfUfuUfuuAfuAfaua-3’ (SEQ ID NO: 2876), and the antisense strand comprises 5’-usAfsUfuAfuaAfaAfauaUfcUfuGfcuususudTdT-3’ (SEQ ID NO: 2889), wherein a, g, c, and u are 2’-O-methyl (2’-OMe) A, G, C, and U, respectively; Af, Gf, Cf, and Uf are 2’-fluoro A, G, C, and U, respectively; dT is deoxy-thymine nucleotide; and s is a phosphorothioate linkage. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 400 mg once a month for 2 months, e.g., prior to the administration of eculizumab. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 400 mg once a month for 2 months, e.g., prior to the administration of eculizumab, and then once every 3 months thereafter. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 400 mg once a month for 2 months, e.g., prior to the administration of eculizumab, and then once every 6 months thereafter. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 400 mg once a month for a long period of time.

[0027] In another aspect, the present invention provides a method for treating, e.g., treating for a long period of time, a subject suffering from paroxysmal nocturnal hemoglobinuria (PNH). The method comprises administering to a subject with PNH who has not received eculizumab a double-stranded ribonucleic acid (dsRNA) agent at a fixed dose of 400 mg once a month for inhibiting the expression of complement component C5; and administering to the subject a dose of about 600 mg of eculizumab or an antigen-binding fragment thereof, thereby treating the subject, wherein the dsRNA agent comprises a sense strand and an antisense strand, wherein the sense strand comprises 5'-asasGfcAfaGfaUfAfUfuUfuuAfuAfaua-3' (SEQ ID NO: 2876), and the antisense strand comprises 5'-usAfsUfuAfuaAfaAfauaUfcUfuGfcuususudTdT-3' (SEQ ID NO: 2889), wherein a, g, c, and u are 2'-O-methyl (2'-OMe) A, G, C, and U, respectively; Af, Gf, Cf, and Uf are 2'-fluoro A, G, C, and U, respectively; dT is deoxy-thymidine nucleotide; and s is a phosphorothioate bond. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 400 mg once a month for 2 months, e.g., prior to the administration of eculizumab. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 400 mg once a month for 2 months and then once every 3 months, e.g., prior to the administration of eculizumab. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 400 mg once a month for 2 months and then once every 6 months, e.g., prior to the administration of eculizumab. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 400 mg once a month for a long period of time.

[0028] In another aspect, the present invention provides a method for treating, e.g., treating over a long period of time, a subject suffering from paroxysmal nocturnal hemoglobinuria (PNH). The method comprises administering to a subject with PNH who has not been administered eculizumab, once a month, a fixed dose of 400 mg of a double-stranded ribonucleic acid (dsRNA) agent for inhibiting the expression of complement component C5; and administering to the subject a dose of about 900 mg of eculizumab, or an antigen-binding fragment thereof, thereby treating the subject, wherein the dsRNA agent comprises a sense strand and an antisense strand, wherein the sense strand comprises 5’-asasGfcAfaGfaUfAfUfuUfuuAfuAfaua-3’ (SEQ ID NO: 2876), and the antisense strand comprises 5’-usAfsUfuAfuaAfaAfauaUfcUfuGfcuususudTdT-3’ (SEQ ID NO: 2889), Here, a, g, c, and u are 2'-O-methyl (2'-OMe) A, G, C, and U, respectively; Af, Gf, Cf, and Uf are 2'-fluoro A, G, C, and U, respectively; dT is deoxy-thymidine nucleotide; and s is a phosphorothioate bond. In one embodiment, the dsRNA agent is administered to a subject at a fixed dose of 400 mg once a month for 2 months, for example, prior to the administration of eculizumab. In one embodiment, the dsRNA agent is administered to a subject at a fixed dose of 400 mg once a month for 2 months, for example, prior to the administration of eculizumab, and then once every 3 months thereafter. In one embodiment, the dsRNA agent is administered to a subject at a fixed dose of 400 mg once a month for 2 months, for example, prior to the administration of eculizumab, and then once every 6 months thereafter. In one embodiment, the dsRNA agent is administered to a subject at a fixed dose of 400 mg once a month for a long period of time. In one aspect, the present invention provides a method for treating, for example, treating for a long period of time, a subject suffering from paroxysmal nocturnal hemoglobinuria (PNH). The method includes administering to a subject suffering from PNH and previously treated with eculizumab a double-stranded ribonucleic acid (dsRNA) agent at a fixed dose of 400 mg once a week to inhibit the expression of complement component C5; and administering to the subject eculizumab or an antigen-binding fragment thereof at a dose of about 300 mg to about 900 mg once every 4 weeks, thereby treating the subject, wherein the dsRNA agent includes a sense strand and an antisense strand, wherein the sense strand includes 5'-asasGfcAfaGfaUfAfUfuUfuuAfuAfaua-3' (SEQ ID NO: 2876), and the antisense strand includes 5'-usAfsUfuAfuaAfaAfauaUfcUfuGfcuususudTdT-3' (SEQ ID NO: 2889), where a, g, c, and u are 2'-O-methyl (2'-OMe) A, G, C, and U, respectively; Af, Gf, Cf, and Uf are 2'-fluoro A, G, C, and U, respectively; dT is deoxy-thymidine nucleotide; and s is a phosphorothioate bond.In one embodiment, the dsRNA agent is administered to a subject once a week for 8 weeks at a fixed dose of 400 mg of the dsRNA agent, for example, before administration of eculizumab at a dose of about 300 mg to about 900 mg, or an antigen-binding fragment thereof. In one embodiment, the dsRNA agent is administered to a subject once a week for 8 weeks at a fixed dose of 400 mg of the dsRNA agent, for example, before administration of eculizumab at a dose of about 300 mg to about 900 mg, or an antigen-binding fragment thereof, and then once a month thereafter. In one embodiment, the dsRNA agent is administered to a subject once a week for 8 weeks at a fixed dose of 400 mg of the dsRNA agent, for example, before administration of eculizumab at a dose of about 300 mg to about 900 mg, or an antigen-binding fragment thereof, and then once every 3 months thereafter. In one embodiment, the dsRNA agent is administered to a subject once a week for 8 weeks at a fixed dose of 400 mg of the dsRNA agent, for example, before administration of eculizumab at a dose of about 300 mg to about 900 mg, or an antigen-binding fragment thereof, and then once every 6 months thereafter. In one embodiment, the dsRNA agent is administered to a subject at a fixed dose of 400 mg of the dsRNA agent once a week for a long period of time.

[0029] In one aspect, the present invention provides a method for treating, e.g., treating for a long period of time, a subject suffering from paroxysmal nocturnal hemoglobinuria (PNH). The method comprises administering to a subject suffering from PNH and previously treated with eculizumab, once a week, a fixed dose of 400 mg of a double-stranded ribonucleic acid (dsRNA) agent for inhibiting the expression of complement component C5; and administering to the subject, once every 4 weeks, a dose of about 300 mg of eculizumab, or an antigen-binding fragment thereof, thereby treating the subject, wherein the dsRNA agent comprises a sense strand and an antisense strand, wherein the sense strand comprises 5'-asasGfcAfaGfaUfAfUfuUfuuAfuAfaua-3' (SEQ ID NO: 2876), and the antisense strand comprises 5'-usAfsUfuAfuaAfaAfauaUfcUfuGfcuususudTdT-3' (SEQ ID NO: 2889), wherein a, g, c, and u are 2'-O-methyl (2'-OMe) A, G, C, and U, respectively; Af, Gf, Cf, and Uf are 2'-fluoro A, G, C, and U, respectively; dT is a deoxy-thymine nucleotide; and s is a phosphorothioate bond. In one embodiment, the dsRNA agent is administered to the subject once a week, at a fixed dose of 400 mg, for 8 weeks prior to the administration of, e.g., a dose of about 300 mg of eculizumab, or an antigen-binding fragment thereof. In one embodiment, the dsRNA agent is administered to the subject once a week, at a fixed dose of 400 mg, for 8 weeks prior to the administration of, e.g., a dose of about 300 mg of eculizumab, or an antigen-binding fragment thereof, and then once a month thereafter. In one embodiment, the dsRNA agent is administered to the subject once a week, at a fixed dose of 400 mg, for 8 weeks prior to the administration of, e.g., a dose of about 300 mg of eculizumab, or an antigen-binding fragment thereof, and then once every 3 months thereafter. In one embodiment, the dsRNA agent is administered to the subject once a week, at a fixed dose of 400 mg, for 8 weeks prior to the administration of, e.g., a dose of about 300 mg of eculizumab, or an antigen-binding fragment thereof, and then once every 6 months thereafter. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 400 mg once a week for a long period of time.

[0030] In one aspect, the present invention provides a method for treating, e.g., treating over a long period of time, a subject suffering from paroxysmal nocturnal hemoglobinuria (PNH). The method comprises administering to a subject suffering from PNH and previously treated with eculizumab, once a week, a fixed dose of 400 mg of a double-stranded ribonucleic acid (dsRNA) agent for inhibiting the expression of complement component C5; and administering to the subject, once every 4 weeks, a dose of about 600 mg of eculizumab, or an antigen-binding fragment thereof, thereby treating the subject, wherein the dsRNA agent comprises a sense strand and an antisense strand, wherein the sense strand comprises 5'-asasGfcAfaGfaUfAfUfuUfuuAfuAfaua-3' (SEQ ID NO: 2876), and the antisense strand comprises 5'-usAfsUfuAfuaAfaAfauaUfcUfuGfcuususudTdT-3' (SEQ ID NO: 2889), wherein a, g, c, and u are 2'-O-methyl (2'-OMe) A, G, C, and U, respectively; Af, Gf, Cf, and Uf are 2'-fluoro A, G, C, and U, respectively; dT is deoxy-thymidine nucleotide; and s is a phosphorothioate bond. In one embodiment, the dsRNA agent is administered to the subject once a week, at a fixed dose of 400 mg, for 8 weeks prior to the administration of, e.g., a dose of about 600 mg of eculizumab, or an antigen-binding fragment thereof. In one embodiment, the dsRNA agent is administered to the subject once a week, at a fixed dose of 400 mg, for 8 weeks prior to the administration of, e.g., a dose of about 600 mg of eculizumab, or an antigen-binding fragment thereof, and then once a month thereafter. In one embodiment, the dsRNA agent is administered to the subject once a week, at a fixed dose of 400 mg, for 8 weeks prior to the administration of, e.g., a dose of about 600 mg of eculizumab, or an antigen-binding fragment thereof, and then once every 3 months thereafter. In one embodiment, the dsRNA agent is administered to the subject once a week, at a fixed dose of 400 mg, for 8 weeks prior to the administration of, e.g., a dose of about 600 mg of eculizumab, or an antigen-binding fragment thereof, and then once every 6 months thereafter. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 400 mg once a week for a long period of time.

[0031] In one aspect, the present invention provides a method for treating, e.g., treating for a long period of time, a subject suffering from paroxysmal nocturnal hemoglobinuria (PNH). The method comprises administering to a subject suffering from PNH and previously treated with eculizumab, once a week, a fixed dose of 400 mg of a double-stranded ribonucleic acid (dsRNA) agent for inhibiting the expression of complement component C5; and administering to the subject, once every 4 weeks, a dose of about 900 mg of eculizumab, or an antigen-binding fragment thereof, thereby treating the subject, wherein the dsRNA agent comprises a sense strand and an antisense strand, wherein the sense strand comprises 5'-asasGfcAfaGfaUfAfUfuUfuuAfuAfaua-3' (SEQ ID NO: 2876), and the antisense strand comprises 5'-usAfsUfuAfuaAfaAfauaUfcUfuGfcuususudTdT-3' (SEQ ID NO: 2889), wherein a, g, c, and u are 2'-O-methyl (2'-OMe) A, G, C, and U, respectively; Af, Gf, Cf, and Uf are 2'-fluoro A, G, C, and U, respectively; dT is a deoxy-thymine nucleotide; and s is a phosphorothioate bond. In one embodiment, the dsRNA agent is administered to the subject once a week for 8 weeks at a fixed dose of 400 mg of the dsRNA agent, e.g., prior to the administration of a dose of about 900 mg of eculizumab, or an antigen-binding fragment thereof. In one embodiment, the dsRNA agent is administered to the subject once a week for 8 weeks at a fixed dose of 400 mg of the dsRNA agent, e.g., prior to the administration of a dose of about 900 mg of eculizumab, or an antigen-binding fragment thereof, and then once a month. In one embodiment, the dsRNA agent is administered to the subject once a week for 8 weeks at a fixed dose of 400 mg of the dsRNA agent, e.g., prior to the administration of a dose of about 900 mg of eculizumab, or an antigen-binding fragment thereof, and then once every 3 months. In one embodiment, the dsRNA agent is administered to the subject once a week for 8 weeks at a fixed dose of 400 mg of the dsRNA agent, e.g., prior to the administration of a dose of about 900 mg of eculizumab, or an antigen-binding fragment thereof, and then once every 6 months. In one embodiment, the dsRNA agent is administered to the subject for a long period of time at a fixed dose of 400 mg of the dsRNA agent once a week.

[0032] In another aspect, the present invention provides a method for treating, e.g., treating for a long period of time, a subject suffering from paroxysmal nocturnal hemoglobinuria (PNH), the method comprising administering, to a subject suffering from PNH and previously treated with eculizumab, a double-stranded ribonucleic acid (dsRNA) agent at a fixed dose of 400 mg once a month to inhibit the expression of complement component C5; and administering to the subject eculizumab or an antigen-binding fragment thereof at a dose of about 300 mg to about 900 mg once every four weeks, thereby treating the subject, wherein the dsRNA agent comprises a sense strand and an antisense strand, wherein the sense strand comprises 5'-asasGfcAfaGfaUfAfUfuUfuuAfuAfaua-3' (SEQ ID NO: 2876), and the antisense strand comprises 5'-usAfsUfuAfuaAfaAfauaUfcUfuGfcuususudTdT-3' (SEQ ID NO: 2889), wherein a, g, c, and u are 2'-O-methyl (2'-OMe) A, G, C, and U, respectively; Af, Gf, Cf, and Uf are 2'-fluoro A, G, C, and U, respectively; dT is deoxy-thymidine nucleotide; and s is a phosphorothioate bond. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 400 mg once a month for two months prior to administration of eculizumab or an antigen-binding fragment thereof at a dose of about 300 mg to about 900 mg. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 400 mg once a month for two months prior to administration of eculizumab or an antigen-binding fragment thereof at a dose of about 300 mg to about 900 mg, and then once every three months thereafter. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 400 mg once a month for two months prior to administration of eculizumab or an antigen-binding fragment thereof at a dose of about 300 mg to about 900 mg, and then once every six months thereafter. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 400 mg once a month for a long period of time.

[0033] In another aspect, the present invention provides a method for treating, e.g., treating over a long period of time, a subject suffering from paroxysmal nocturnal hemoglobinuria (PNH), the method comprising administering to a subject suffering from PNH and previously treated with eculizumab, once a month, a fixed dose of 400 mg of a double-stranded ribonucleic acid (dsRNA) agent for inhibiting the expression of complement component C5; and administering to the subject, once every four weeks, a dose of about 300 mg of eculizumab, or an antigen-binding fragment thereof, thereby treating the subject, wherein the dsRNA agent comprises a sense strand and an antisense strand, wherein the sense strand comprises 5'-asasGfcAfaGfaUfAfUfuUfuuAfuAfaua-3' (SEQ ID NO: 2876), and the antisense strand comprises 5'-usAfsUfuAfuaAfaAfauaUfcUfuGfcuususudTdT-3' (SEQ ID NO: 2889), wherein a, g, c, and u are 2'-O-methyl (2'-OMe) A, G, C, and U, respectively; Af, Gf, Cf, and Uf are 2'-fluoro A, G, C, and U, respectively; dT is a deoxy-thymine nucleotide; and s is a phosphorothioate linkage. In one embodiment, the dsRNA agent is administered to the subject once a month, at a fixed dose of 400 mg, for two months prior to, e.g., administration of a dose of about 300 mg of eculizumab, or an antigen-binding fragment thereof. In one embodiment, the dsRNA agent is administered to the subject once a month, at a fixed dose of 400 mg, for two months prior to, e.g., administration of a dose of about 300 mg of eculizumab, or an antigen-binding fragment thereof, and then once every three months. In one embodiment, the dsRNA agent is administered to the subject once a month, at a fixed dose of 400 mg, for two months prior to, e.g., administration of a dose of about 300 mg of eculizumab, or an antigen-binding fragment thereof, and then once every six months. In one embodiment, the dsRNA agent is administered to the subject over a long period of time, once a month, at a fixed dose of 400 mg.

[0034] In another aspect, the present invention provides a method for treating, e.g., treating over a long period of time, a subject suffering from paroxysmal nocturnal hemoglobinuria (PNH), the method comprising administering to a subject suffering from PNH and previously treated with eculizumab a double-stranded ribonucleic acid (dsRNA) agent at a fixed dose of 400 mg once a month to inhibit the expression of complement component C5; and administering to the subject eculizumab or an antigen-binding fragment thereof at a dose of about 600 mg once every four weeks, thereby treating the subject, wherein the dsRNA agent comprises a sense strand and an antisense strand, wherein the sense strand comprises 5'-asasGfcAfaGfaUfAfUfuUfuuAfuAfaua-3' (SEQ ID NO: 2876), and the antisense strand comprises 5'-usAfsUfuAfuaAfaAfauaUfcUfuGfcuususudTdT-3' (SEQ ID NO: 2889), wherein a, g, c, and u are 2'-O-methyl (2'-OMe) A, G, C, and U, respectively; Af, Gf, Cf, and Uf are 2'-fluoro A, G, C, and U, respectively; dT is deoxy-thymidine nucleotide; and s is a phosphorothioate bond. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 400 mg once a month for two months prior to, e.g., administration of eculizumab or an antigen-binding fragment thereof at a dose of about 600 mg. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 400 mg once a month for two months prior to, e.g., administration of eculizumab or an antigen-binding fragment thereof at a dose of about 600 mg, and then once every three months thereafter. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 400 mg once a month for two months prior to, e.g., administration of eculizumab or an antigen-binding fragment thereof at a dose of about 600 mg, and then once every six months thereafter. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 400 mg once a month for a long period of time.

[0035] In another aspect, the present invention provides a method for treating, e.g., treating for a long period of time, a subject suffering from paroxysmal nocturnal hemoglobinuria (PNH), the method comprising administering to a subject suffering from PNH and previously treated with eculizumab, a double-stranded ribonucleic acid (dsRNA) agent that inhibits the expression of complement component C5 at a fixed dose of 400 mg once a month; and administering to the subject eculizumab or an antigen-binding fragment thereof at a dose of about 900 mg once every four weeks, thereby treating the subject, wherein the dsRNA agent comprises a sense strand and an antisense strand, wherein the sense strand comprises 5’-asasGfcAfaGfaUfAfUfuUfuuAfuAfaua-3’ (SEQ ID NO: 2876), and the antisense strand comprises 5’-usAfsUfuAfuaAfaAfauaUfcUfuGfcuususudTdT-3’ (SEQ ID NO: 2889), wherein a, g, c, and u are 2’-O-methyl (2’-OMe) A, G, C, and U, respectively; Af, Gf, Cf, and Uf are 2’-fluoro A, G, C, and U, respectively; dT is deoxy-thymidine nucleotide; and s is a phosphorothioate bond. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 400 mg once a month for two months prior to, e.g., the administration of eculizumab or an antigen-binding fragment thereof at a dose of about 900 mg. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 400 mg once a month for two months prior to, and then once every three months after, the administration of eculizumab or an antigen-binding fragment thereof at a dose of about 300 mg. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 400 mg once a month for two months prior to, and then once every six months after, the administration of eculizumab or an antigen-binding fragment thereof at a dose of about 300 mg. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 400 mg once a month for a long period of time. In one aspect, the present invention provides a method for treating, e.g., treating for a long period of time, a subject suffering from paroxysmal nocturnal hemoglobinuria (PNH).This method involves administering to a subject suffering from PNH and previously treated with eculizumab a double-stranded ribonucleic acid (dsRNA) agent at a fixed dose of 400 mg once a week for 2 to 8 weeks to inhibit the expression of complement component C5; and administering to the subject eculizumab or an antigen-binding fragment thereof at a dose of about 300 mg to about 900 mg once every 4 weeks, thereby treating the subject, wherein the dsRNA agent comprises a sense strand and an antisense strand, wherein the sense strand comprises 5'-asasGfcAfaGfaUfAfUfuUfuuAfuAfaua-3' (SEQ ID NO: 2876), the antisense strand comprises 5'-usAfsUfuAfuaAfaAfauaUfcUfuGfcuususudTdT-3' (SEQ ID NO: 2889), wherein a, g, c, and u are 2'-O-methyl (2'-OMe) A, G, C, and U, respectively; Af, Gf, Cf, and Uf are 2'-fluoro A, G, C, and U, respectively; dT is deoxy-thymidine nucleotide; and s is a phosphorothioate bond. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 400 mg once a week for 2 to 8 weeks before administering to the subject eculizumab or an antigen-binding fragment thereof at a dose of about 300 mg to about 900 mg. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 400 mg once a week for 2 to 8 weeks before administering to the subject eculizumab or an antigen-binding fragment thereof at a dose of about 300 mg to about 900 mg, and then once a month thereafter. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 400 mg once a week for 2 to 8 weeks before administering to the subject eculizumab or an antigen-binding fragment thereof at a dose of about 300 mg to about 900 mg, and then once every 3 months thereafter. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 400 mg once a week for 2 to 8 weeks before administering to the subject eculizumab or an antigen-binding fragment thereof at a dose of about 300 mg to about 900 mg, and then once every 6 months thereafter. In one embodiment, the dsRNA agent is administered to the subject for a long period of time as a dsRNA agent at a fixed dose of 400 mg once a week.

[0036] In one aspect, the present invention provides a method for treating, e.g., treating for a long period of time, a subject suffering from paroxysmal nocturnal hemoglobinuria (PNH). The method comprises administering to a subject suffering from PNH and previously treated with eculizumab, once a week for 2 to 8 weeks, a fixed dose of 400 mg of a double-stranded ribonucleic acid (dsRNA) agent for inhibiting the expression of complement component C5; and administering to the subject once every 4 weeks, a dose of about 300 mg of eculizumab, or an antigen-binding fragment thereof, thereby treating the subject, wherein the dsRNA agent comprises a sense strand and an antisense strand, wherein the sense strand comprises 5’-asasGfcAfaGfaUfAfUfuUfuuAfuAfaua-3’ (SEQ ID NO: 2876), and the antisense strand comprises 5’-usAfsUfuAfuaAfaAfauaUfcUfuGfcuususudTdT-3’ (SEQ ID NO: 2889), wherein a, g, c, and u are 2’-O-methyl (2’-OMe) A, G, C, and U, respectively; Af, Gf, Cf, and Uf are 2’-fluoro A, G, C, and U, respectively; dT is a deoxy-thymine nucleotide; and s is a phosphorothioate bond. In one embodiment, the dsRNA agent is administered to the subject once a week for 2 to 8 weeks at a fixed dose of 400 mg prior to administration of a dose of about 300 mg of eculizumab, or an antigen-binding fragment thereof, to the subject. In one embodiment, the dsRNA agent is administered to the subject once a week for 2 to 8 weeks at a fixed dose of 400 mg prior to administration of a dose of about 300 mg of eculizumab, or an antigen-binding fragment thereof, to the subject and then once a month thereafter. In one embodiment, the dsRNA agent is administered to the subject once a week for 2 to 8 weeks at a fixed dose of 400 mg prior to administration of a dose of about 300 mg of eculizumab, or an antigen-binding fragment thereof, to the subject and then once every 3 months thereafter. In one embodiment, the dsRNA agent is administered to the subject once a week for 2 to 8 weeks at a fixed dose of 400 mg prior to administration of a dose of about 300 mg of eculizumab, or an antigen-binding fragment thereof, to the subject and then once every 6 months thereafter. In one embodiment, the dsRNA agent is administered to the subject for a long period of time at a fixed dose of 400 mg of the dsRNA agent once a week.

[0037] In one aspect, the present invention provides a method for treating, e.g., treating over a long period of time, a subject suffering from paroxysmal nocturnal hemoglobinuria (PNH). The method comprises administering to a subject suffering from PNH and previously treated with eculizumab, once a week for 2 to 8 weeks, a fixed dose of 400 mg of a double-stranded ribonucleic acid (dsRNA) agent for inhibiting the expression of complement component C5; and administering to the subject once every 4 weeks a dose of about 600 mg of eculizumab, or an antigen-binding fragment thereof, thereby treating the subject, wherein the dsRNA agent comprises a sense strand and an antisense strand, wherein the sense strand comprises 5'-asasGfcAfaGfaUfAfUfuUfuuAfuAfaua-3' (SEQ ID NO: 2876), and the antisense strand comprises 5'-usAfsUfuAfuaAfaAfauaUfcUfuGfcuususudTdT-3' (SEQ ID NO: 2889), wherein a, g, c, and u are 2'-O-methyl (2'-OMe) A, G, C, and U, respectively; Af, Gf, Cf, and Uf are 2'-fluoro A, G, C, and U, respectively; dT is deoxy-thymidine nucleotide; and s is a phosphorothioate linkage. In one embodiment, the dsRNA agent is administered to the subject once a week for 2 to 8 weeks at a fixed dose of 400 mg prior to administration of a dose of about 600 mg of eculizumab, or an antigen-binding fragment thereof, to the subject. In one embodiment, the dsRNA agent is administered to the subject once a week for 2 to 8 weeks at a fixed dose of 400 mg prior to administration of a dose of about 600 mg of eculizumab, or an antigen-binding fragment thereof, to the subject, and then once a month thereafter. In one embodiment, the dsRNA agent is administered to the subject once a week for 2 to 8 weeks at a fixed dose of 400 mg prior to administration of a dose of about 600 mg of eculizumab, or an antigen-binding fragment thereof, to the subject, and then once every 3 months thereafter. In one embodiment, the dsRNA agent is administered to the subject once a week for 2 to 8 weeks at a fixed dose of 400 mg prior to administration of a dose of about 600 mg of eculizumab, or an antigen-binding fragment thereof, to the subject, and then once every 6 months thereafter. In one embodiment, the dsRNA agent is administered to the subject over a long period of time at a fixed dose of 400 mg of the dsRNA agent once a week.

[0038] In one aspect, the present invention provides a method for treating, e.g., treating over a long period of time, a subject suffering from paroxysmal nocturnal hemoglobinuria (PNH). The method comprises administering to a subject suffering from PNH and previously treated with eculizumab, once a week for 2 to 8 weeks, a fixed dose of 400 mg of a double-stranded ribonucleic acid (dsRNA) agent for inhibiting the expression of complement component C5; and administering to the subject, once every 4 weeks, a dose of about 900 mg of eculizumab, or an antigen-binding fragment thereof, thereby treating the subject, wherein the dsRNA agent comprises a sense strand and an antisense strand, wherein the sense strand comprises 5'-asasGfcAfaGfaUfAfUfuUfuuAfuAfaua-3' (SEQ ID NO: 2876), and the antisense strand comprises 5'-usAfsUfuAfuaAfaAfauaUfcUfuGfcuususudTdT-3' (SEQ ID NO: 2889), wherein a, g, c, and u are 2'-O-methyl (2'-OMe) A, G, C, and U, respectively; Af, Gf, Cf, and Uf are 2'-fluoro A, G, C, and U, respectively; dT is a deoxy-thymine nucleotide; and s is a phosphorothioate bond. In one embodiment, the dsRNA agent is administered to the subject once a week for 2 to 8 weeks at a fixed dose of 400 mg prior to administration of a dose of about 900 mg of eculizumab, or an antigen-binding fragment thereof, to the subject. In one embodiment, the dsRNA agent is administered to the subject once a week for 2 to 8 weeks at a fixed dose of 400 mg prior to administration of a dose of about 900 mg of eculizumab, or an antigen-binding fragment thereof, to the subject, and then once a month thereafter. In one embodiment, the dsRNA agent is administered to the subject once a week for 2 to 8 weeks at a fixed dose of 400 mg prior to administration of a dose of about 900 mg of eculizumab, or an antigen-binding fragment thereof, to the subject, and then once every 3 months thereafter. In one embodiment, the dsRNA agent is administered to the subject once a week for 2 to 8 weeks at a fixed dose of 400 mg prior to administration of a dose of about 900 mg of eculizumab, or an antigen-binding fragment thereof, to the subject, and then once every 6 months thereafter. In one embodiment, the dsRNA agent is administered to the subject over a long period of time at a fixed dose of 400 mg of the dsRNA agent once a week.In another aspect, the present invention provides a method for treating, e.g., treating over a long period of time, a subject suffering from paroxysmal nocturnal hemoglobinuria (PNH). The method comprises administering to a subject suffering from PNH and previously treated with eculizumab, a double-stranded ribonucleic acid (dsRNA) agent at a fixed dose of 400 mg once a month for 1 to 2 months to inhibit the expression of complement component C5; and administering to the subject eculizumab or an antigen-binding fragment thereof at a dose of about 300 mg to about 900 mg once every 4 weeks, thereby treating the subject, wherein the dsRNA agent comprises a sense strand and an antisense strand, wherein the sense strand comprises 5’-asasGfcAfaGfaUfAfUfuUfuuAfuAfaua-3’ (SEQ ID NO: 2876), and the antisense strand comprises 5’-usAfsUfuAfuaAfaAfauaUfcUfuGfcuususudTdT-3’ (SEQ ID NO: 2889), wherein a, g, c, and u are 2’-O-methyl (2’-OMe) A, G, C, and U, respectively; Af, Gf, Cf, and Uf are 2’-fluoro A, G, C, and U, respectively; dT is deoxy-thymidine nucleotide; and s is a phosphorothioate linkage. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 400 mg once a month for 1 to 2 months prior to administration of eculizumab or an antigen-binding fragment thereof to the subject at a dose of about 300 mg to about 900 mg. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 400 mg once a month for 1 to 2 months prior to administration of eculizumab or an antigen-binding fragment thereof to the subject at a dose of about 300 mg to about 900 mg, and then once every 3 months. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 400 mg once a month for 1 to 2 months prior to administration of eculizumab or an antigen-binding fragment thereof to the subject at a dose of about 300 mg to about 900 mg, and then once every 6 months. In one embodiment, the dsRNA agent is administered to the subject over a long period of time at a fixed dose of 400 mg of the dsRNA agent once a month.

[0039] In another aspect, the present invention provides a method for treating, e.g., treating over a long period of time, a subject suffering from paroxysmal nocturnal hemoglobinuria (PNH). The method comprises administering to a subject suffering from PNH and previously treated with eculizumab, a double-stranded ribonucleic acid (dsRNA) agent at a fixed dose of 400 mg once a month for 1 to 2 months to inhibit the expression of complement component C5; and administering to the subject eculizumab, or an antigen-binding fragment thereof, at a dose of about 300 mg once every 4 weeks, thereby treating the subject, wherein the dsRNA agent comprises a sense strand and an antisense strand, wherein the sense strand comprises 5'-asasGfcAfaGfaUfAfUfuUfuuAfuAfaua-3' (SEQ ID NO: 2876), and the antisense strand comprises 5'-usAfsUfuAfuaAfaAfauaUfcUfuGfcuususudTdT-3' (SEQ ID NO: 2889), wherein a, g, c, and u are 2'-O-methyl (2'-OMe) A, G, C, and U, respectively; Af, Gf, Cf, and Uf are 2'-fluoro A, G, C, and U, respectively; dT is deoxy-thymidine nucleotide; and s is a phosphorothioate bond. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 400 mg once a month for 1 to 2 months prior to administration of eculizumab, or an antigen-binding fragment thereof, to the subject at a dose of about 300 mg. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 400 mg once a month for 1 to 2 months prior to administration of eculizumab, or an antigen-binding fragment thereof, to the subject at a dose of about 300 mg, and then once every 3 months thereafter. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 400 mg once a month for 1 to 2 months prior to administration of eculizumab, or an antigen-binding fragment thereof, to the subject at a dose of about 300 mg, and then once every 6 months thereafter. In one embodiment, the dsRNA agent is administered to the subject over a long period of time at a fixed dose of 400 mg of the dsRNA agent once a month.

[0040] In another aspect, the present invention provides a method for treating, e.g., treating over a long period of time, a subject suffering from paroxysmal nocturnal hemoglobinuria (PNH). The method comprises administering to a subject suffering from PNH and previously treated with eculizumab, a double-stranded ribonucleic acid (dsRNA) agent at a fixed dose of 400 mg once a month for 1 to 2 months to inhibit the expression of complement component C5; and administering to the subject eculizumab or an antigen-binding fragment thereof at a dose of about 600 mg once every 4 weeks, thereby treating the subject, wherein the dsRNA agent comprises a sense strand and an antisense strand, wherein the sense strand comprises 5'-asasGfcAfaGfaUfAfUfuUfuuAfuAfaua-3' (SEQ ID NO: 2876), and the antisense strand comprises 5'-usAfsUfuAfuaAfaAfauaUfcUfuGfcuususudTdT-3' (SEQ ID NO: 2889), wherein a, g, c, and u are 2'-O-methyl (2'-OMe) A, G, C, and U, respectively; Af, Gf, Cf, and Uf are 2'-fluoro A, G, C, and U, respectively; dT is deoxy-thymidine nucleotide; and s is a phosphorothioate bond. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 400 mg once a month for 1 to 2 months prior to administration of eculizumab or an antigen-binding fragment thereof to the subject at a dose of about 600 mg. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 400 mg once a month for 1 to 2 months prior to administration of eculizumab or an antigen-binding fragment thereof to the subject at a dose of about 600 mg, and then once every 3 months thereafter. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 400 mg once a month for 1 to 2 months prior to administration of eculizumab or an antigen-binding fragment thereof to the subject at a dose of about 600 mg, and then once every 6 months thereafter. In one embodiment, the dsRNA agent is administered to the subject over a long period of time at a fixed dose of 400 mg of the dsRNA agent once a month.

[0041] In another aspect, the present invention provides a method for treating, e.g., treating over a long period of time, a subject suffering from paroxysmal nocturnal hemoglobinuria (PNH). The method comprises administering to a subject suffering from PNH and previously treated with eculizumab, a double-stranded ribonucleic acid (dsRNA) agent at a fixed dose of 400 mg once a month for 1 to 2 months to inhibit the expression of complement component C5; and administering to the subject eculizumab, or an antigen-binding fragment thereof, at a dose of about 900 mg once every 4 weeks, thereby treating the subject, wherein the dsRNA agent comprises a sense strand and an antisense strand, wherein the sense strand comprises 5’-asasGfcAfaGfaUfAfUfuUfuuAfuAfaua-3’ (SEQ ID NO: 2876), and the antisense strand comprises 5’-usAfsUfuAfuaAfaAfauaUfcUfuGfcuususudTdT-3’ (SEQ ID NO: 2889), wherein a, g, c, and u are 2’-O-methyl (2’-OMe) A, G, C, and U, respectively; Af, Gf, Cf, and Uf are 2’-fluoro A, G, C, and U, respectively; dT is deoxy-thymidine nucleotide; and s is a phosphorothioate bond. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 400 mg once a month for 1 to 2 months prior to administration of eculizumab, or an antigen-binding fragment thereof, to the subject at a dose of about 300 mg. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 400 mg once a month for 1 to 2 months prior to administration of eculizumab, or an antigen-binding fragment thereof, to the subject at a dose of about 300 mg, and then once every 3 months thereafter. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 400 mg once a month for 1 to 2 months prior to administration of eculizumab, or an antigen-binding fragment thereof, to the subject at a dose of about 300 mg, and then once every 6 months thereafter. In one embodiment, the dsRNA agent is administered to the subject over a long period of time at a fixed dose of 400 mg of the dsRNA agent once a month.

[0042] In one aspect, the present invention provides a method for treating, e.g., treating over a long period of time, a subject suffering from paroxysmal nocturnal hemoglobinuria (PNH). The method comprises administering to a subject suffering from PNH and previously treated with eculizumab, once a week, a fixed dose of 200 mg of a double-stranded ribonucleic acid (dsRNA) agent for inhibiting the expression of complement component C5; and administering to the subject, once every 4 weeks, a dose of eculizumab of about 300 mg to about 900 mg, or an antigen-binding fragment thereof, thereby treating the subject, wherein the dsRNA agent comprises a sense strand and an antisense strand, wherein the sense strand comprises 5’-asasGfcAfaGfaUfAfUfuUfuuAfuAfaua-3’ (SEQ ID NO: 2876), and the antisense strand comprises 5’-usAfsUfuAfuaAfaAfauaUfcUfuGfcuususudTdT-3’ (SEQ ID NO: 2889), wherein a, g, c, and u are 2’-O-methyl (2’-OMe) A, G, C, and U, respectively; Af, Gf, Cf, and Uf are 2’-fluoro A, G, C, and U, respectively; dT is a deoxy-thymine nucleotide; and s is a phosphorothioate bond. In one embodiment, the dsRNA agent is administered to the subject once a week at a fixed dose of 200 mg for 8 weeks prior to administration of, e.g., a dose of eculizumab of about 300 mg to about 900 mg, or an antigen-binding fragment thereof, to the subject. In another embodiment, the dsRNA agent is administered to the subject once a week at a fixed dose of 200 mg for 12 weeks prior to administration of, e.g., a dose of eculizumab of about 300 mg to about 900 mg, or an antigen-binding fragment thereof, to the subject. In one embodiment, the dsRNA agent is administered to the subject once a week at a fixed dose of 200 mg for 8 weeks prior to administration of, e.g., a dose of eculizumab of about 300 mg to about 900 mg, or an antigen-binding fragment thereof, to the subject and then once a month thereafter. In one embodiment, the dsRNA agent is administered to the subject once a week at a fixed dose of 200 mg for 8 weeks prior to administration of, e.g., a dose of eculizumab of about 300 mg to about 900 mg, or an antigen-binding fragment thereof, to the subject and then once every 3 months thereafter.In one embodiment, the dsRNA agent is administered to the subject once a week for 8 weeks at a fixed dose of 200 mg of the dsRNA agent, and then once every 6 months, prior to the administration of eculizumab at a dose of, for example, about 300 mg to about 900 mg, or an antigen-binding fragment thereof. In one embodiment, the dsRNA agent is administered to the subject once a week for 12 weeks at a fixed dose of 200 mg of the dsRNA agent, and then once a month, prior to the administration of eculizumab at a dose of, for example, about 300 mg to about 900 mg, or an antigen-binding fragment thereof. In one embodiment, the dsRNA agent is administered to the subject once a week for 12 weeks at a fixed dose of 200 mg of the dsRNA agent, and then once every 3 months, prior to the administration of eculizumab at a dose of, for example, about 300 mg to about 900 mg, or an antigen-binding fragment thereof. In one embodiment, the dsRNA agent is administered to the subject once a week for 12 weeks at a fixed dose of 200 mg of the dsRNA agent, and then once every 6 months, prior to the administration of eculizumab at a dose of, for example, about 300 mg to about 900 mg, or an antigen-binding fragment thereof. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 200 mg of the dsRNA agent once a week for a long period of time.

[0043] In one aspect, the present invention provides a method for treating, e.g., treating over a long period of time, a subject suffering from paroxysmal nocturnal hemoglobinuria (PNH). The method comprises administering to a subject suffering from PNH and previously treated with eculizumab, once a week, a fixed dose of 200 mg of a double-stranded ribonucleic acid (dsRNA) agent for inhibiting the expression of complement component C5; and administering to the subject, once every 4 weeks, a dose of about 300 mg of eculizumab, or an antigen-binding fragment thereof, thereby treating the subject, wherein the dsRNA agent comprises a sense strand and an antisense strand, wherein the sense strand comprises 5'-asasGfcAfaGfaUfAfUfuUfuuAfuAfaua-3' (SEQ ID NO: 2876), and the antisense strand comprises 5'-usAfsUfuAfuaAfaAfauaUfcUfuGfcuususudTdT-3' (SEQ ID NO: 2889), wherein a, g, c, and u are 2'-O-methyl (2'-OMe) A, G, C, and U, respectively; Af, Gf, Cf, and Uf are 2'-fluoro A, G, C, and U, respectively; dT is a deoxy-thymine nucleotide; and s is a phosphorothioate bond. In one embodiment, the dsRNA agent is administered to the subject once a week at a fixed dose of 200 mg for 8 weeks, e.g., prior to administration of a dose of about 300 mg of eculizumab, or an antigen-binding fragment thereof, to the subject. In another embodiment, the dsRNA agent is administered to the subject once a week at a fixed dose of 200 mg for 12 weeks, e.g., prior to administration of a dose of about 300 mg of eculizumab, or an antigen-binding fragment thereof, to the subject. In one embodiment, the dsRNA agent is administered to the subject once a week at a fixed dose of 200 mg for 8 weeks, e.g., prior to administration of a dose of about 300 mg of eculizumab, or an antigen-binding fragment thereof, to the subject, and then once a month thereafter. In one embodiment, the dsRNA agent is administered to the subject once a week at a fixed dose of 200 mg for 8 weeks, e.g., prior to administration of a dose of about 300 mg of eculizumab, or an antigen-binding fragment thereof, to the subject, and then once every 3 months thereafter.In one embodiment, the dsRNA agent is administered to a subject once a week for 8 weeks at a fixed dose of 200 mg of the dsRNA agent, and then once every 6 months, for example, prior to administration of a dose of about 300 mg of eculizumab or an antigen-binding fragment thereof. In one embodiment, the dsRNA agent is administered to a subject once a week for 12 weeks at a fixed dose of 200 mg of the dsRNA agent, and then once a month, for example, prior to administration of a dose of about 300 mg of eculizumab or an antigen-binding fragment thereof. In one embodiment, the dsRNA agent is administered to a subject once a week for 12 weeks at a fixed dose of 200 mg of the dsRNA agent, and then once every 3 months, for example, prior to administration of a dose of about 300 mg of eculizumab or an antigen-binding fragment thereof. In one embodiment, the dsRNA agent is administered to a subject once a week for 12 weeks at a fixed dose of 200 mg of the dsRNA agent, and then once every 6 months, for example, prior to administration of a dose of about 300 mg of eculizumab or an antigen-binding fragment thereof. In one embodiment, the dsRNA agent is administered to a subject at a fixed dose of 200 mg of the dsRNA agent once a week for a long period of time.

[0044] In one aspect, the present invention provides a method for treating, e.g., treating over a long period of time, a subject suffering from paroxysmal nocturnal hemoglobinuria (PNH). The method comprises administering to a subject suffering from PNH and previously treated with eculizumab, once a week, a fixed dose of 200 mg of a double-stranded ribonucleic acid (dsRNA) agent for inhibiting the expression of complement component C5; and administering to the subject, once every 4 weeks, a dose of about 600 mg of eculizumab, or an antigen-binding fragment thereof, thereby treating the subject, wherein the dsRNA agent comprises a sense strand and an antisense strand, wherein the sense strand comprises 5'-asasGfcAfaGfaUfAfUfuUfuuAfuAfaua-3' (SEQ ID NO: 2876), and the antisense strand comprises 5'-usAfsUfuAfuaAfaAfauaUfcUfuGfcuususudTdT-3' (SEQ ID NO: 2889), wherein a, g, c, and u are 2'-O-methyl (2'-OMe) A, G, C, and U, respectively; Af, Gf, Cf, and Uf are 2'-fluoro A, G, C, and U, respectively; dT is a deoxy-thymine nucleotide; and s is a phosphorothioate bond. In one embodiment, the dsRNA agent is administered to the subject once a week at a fixed dose of 200 mg for 8 weeks, e.g., prior to administration of a dose of about 600 mg of eculizumab, or an antigen-binding fragment thereof, to the subject. In another embodiment, the dsRNA agent is administered to the subject once a week at a fixed dose of 200 mg for 12 weeks, e.g., prior to administration of a dose of about 600 mg of eculizumab, or an antigen-binding fragment thereof, to the subject. In one embodiment, the dsRNA agent is administered to the subject once a week at a fixed dose of 200 mg for 8 weeks, e.g., prior to administration of a dose of about 600 mg of eculizumab, or an antigen-binding fragment thereof, to the subject, and then once a month thereafter. In one embodiment, the dsRNA agent is administered to the subject once a week at a fixed dose of 200 mg for 8 weeks, e.g., prior to administration of a dose of about 600 mg of eculizumab, or an antigen-binding fragment thereof, to the subject, and then once every 3 months thereafter.In one embodiment, the dsRNA agent is administered to the subject once a week for 8 weeks at a fixed dose of 200 mg of the dsRNA agent, and then once every 6 months, for example, before administration of a dose of about 600 mg of eculizumab, or an antigen-binding fragment thereof. In one embodiment, the dsRNA agent is administered to the subject once a week for 12 weeks at a fixed dose of 200 mg of the dsRNA agent, and then once a month, for example, before administration of a dose of about 600 mg of eculizumab, or an antigen-binding fragment thereof. In one embodiment, the dsRNA agent is administered to the subject once a week for 12 weeks at a fixed dose of 200 mg of the dsRNA agent, and then once every 3 months, for example, before administration of a dose of about 600 mg of eculizumab, or an antigen-binding fragment thereof. In one embodiment, the dsRNA agent is administered to the subject once a week for 12 weeks at a fixed dose of 200 mg of the dsRNA agent, and then once every 6 months, for example, before administration of a dose of about 600 mg of eculizumab, or an antigen-binding fragment thereof. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 200 mg of the dsRNA agent once a week for a long period of time.

[0045] In one aspect, the present invention provides a method for treating, e.g., treating over a long period of time, a subject suffering from paroxysmal nocturnal hemoglobinuria (PNH). The method comprises administering to a subject suffering from PNH and previously treated with eculizumab, once a week, a fixed dose of 200 mg of a double-stranded ribonucleic acid (dsRNA) agent for inhibiting the expression of complement component C5; and administering to the subject, once every 4 weeks, a dose of about 900 mg of eculizumab, or an antigen-binding fragment thereof, thereby treating the subject, wherein the dsRNA agent comprises a sense strand and an antisense strand, wherein the sense strand comprises 5’-asasGfcAfaGfaUfAfUfuUfuuAfuAfaua-3’ (SEQ ID NO: 2876), and the antisense strand comprises 5’-usAfsUfuAfuaAfaAfauaUfcUfuGfcuususudTdT-3’ (SEQ ID NO: 2889), wherein a, g, c, and u are 2’-O-methyl (2’-OMe) A, G, C, and U, respectively; Af, Gf, Cf, and Uf are 2’-fluoro A, G, C, and U, respectively; dT is a deoxy-thymine nucleotide; and s is a phosphorothioate bond. In one embodiment, the dsRNA agent is administered to the subject once a week, at a fixed dose of 200 mg, for 8 weeks, e.g., prior to administration of a dose of about 900 mg of eculizumab, or an antigen-binding fragment thereof, to the subject. In another embodiment, the dsRNA agent is administered to the subject once a week, at a fixed dose of 200 mg, for 12 weeks, e.g., prior to administration of a dose of about 900 mg of eculizumab, or an antigen-binding fragment thereof, to the subject. In one embodiment, the dsRNA agent is administered to the subject once a week, at a fixed dose of 200 mg, for 8 weeks, e.g., prior to administration of a dose of about 900 mg of eculizumab, or an antigen-binding fragment thereof, to the subject, and then once a month thereafter. In one embodiment, the dsRNA agent is administered to the subject once a week, at a fixed dose of 200 mg, for 8 weeks, e.g., prior to administration of a dose of about 900 mg of eculizumab, or an antigen-binding fragment thereof, to the subject, and then once every 3 months thereafter.In one embodiment, the dsRNA agent is administered to the subject once a week for 8 weeks at a fixed dose of 200 mg of the dsRNA agent, and then once every 6 months, for example, before administration of a dose of about 900 mg of eculizumab, or an antigen-binding fragment thereof. In one embodiment, the dsRNA agent is administered to the subject once a week for 12 weeks at a fixed dose of 200 mg of the dsRNA agent, and then once a month, for example, before administration of a dose of about 900 mg of eculizumab, or an antigen-binding fragment thereof. In one embodiment, the dsRNA agent is administered to the subject once a week for 12 weeks at a fixed dose of 200 mg of the dsRNA agent, and then once every 3 months, for example, before administration of a dose of about 900 mg of eculizumab, or an antigen-binding fragment thereof. In one embodiment, the dsRNA agent is administered to the subject once a week for 12 weeks at a fixed dose of 200 mg of the dsRNA agent, and then once every 6 months, for example, before administration of a dose of about 900 mg of eculizumab, or an antigen-binding fragment thereof. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 200 mg once a week for a long period of time.

[0046] In another aspect, the present invention provides a method for treating, e.g., treating for a long period of time, a subject suffering from paroxysmal nocturnal hemoglobinuria (PNH). The method comprises administering to a subject suffering from PNH and previously treated with eculizumab, once a month, a fixed dose of 200 mg of a double-stranded ribonucleic acid (dsRNA) agent for inhibiting the expression of complement component C5; and administering to the subject, once every 4 weeks, a dose of eculizumab, or an antigen-binding fragment thereof, of about 300 mg to about 900 mg, thereby treating the subject, wherein the dsRNA agent comprises a sense strand and an antisense strand, wherein the sense strand comprises 5'-asasGfcAfaGfaUfAfUfuUfuuAfuAfaua-3' (SEQ ID NO: 2876), the antisense strand comprises 5'-usAfsUfuAfuaAfaAfauaUfcUfuGfcuususudTdT-3' (SEQ ID NO: 2889), wherein a, g, c and u are 2'-O-methyl (2'-OMe) A, G, C, and U, respectively; Af, Gf, Cf and Uf are 2'-fluoro A, G, C and U, respectively; dT is deoxy-thymidine nucleotide; and s is a phosphorothioate bond. In one embodiment, the dsRNA agent is administered to the subject, once a month for 2 months, at a fixed dose of 200 mg of the dsRNA agent, e.g., prior to administration of a dose of eculizumab, or an antigen-binding fragment thereof, of about 300 mg to about 900 mg to the subject. In one embodiment, the dsRNA agent is administered to the subject, once a month for 2 months, at a fixed dose of 200 mg of the dsRNA agent, e.g., prior to administration of a dose of eculizumab, or an antigen-binding fragment thereof, of about 300 mg to about 900 mg to the subject, and then once every 3 months. In one embodiment, the dsRNA agent is administered to the subject, once a month for 2 months, at a fixed dose of 200 mg of the dsRNA agent, e.g., prior to administration of a dose of eculizumab, or an antigen-binding fragment thereof, of about 300 mg to about 900 mg to the subject, and then once every 6 months. In another embodiment, the dsRNA agent is administered to the subject, once a month for 3 months, at a fixed dose of 200 mg of the dsRNA agent, e.g., prior to administration of a dose of eculizumab, or an antigen-binding fragment thereof, of about 300 mg to about 900 mg to the subject.In one embodiment, the dsRNA agent is administered to the subject, for example, at a fixed dose of 200 mg of the dsRNA agent once a month for 3 months, and then once every 3 months, prior to administration of eculizumab, or an antigen-binding fragment thereof, at a dose of about 300 mg to about 900 mg. In one embodiment, the dsRNA agent is administered to the subject, for example, at a fixed dose of 200 mg of the dsRNA agent once a month for 3 months, and then once every 6 months, prior to administration of eculizumab, or an antigen-binding fragment thereof, at a dose of about 300 mg to about 900 mg. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 200 mg of the dsRNA agent once a month for a long period of time.

[0047] In another aspect, the present invention provides a method for treating, e.g., treating over a long period of time, a subject suffering from paroxysmal nocturnal hemoglobinuria (PNH). The method comprises administering to a subject suffering from PNH and previously treated with eculizumab, once a month, a fixed dose of 200 mg of a double-stranded ribonucleic acid (dsRNA) agent for inhibiting the expression of complement component C5; and administering to the subject, once every 4 weeks, a dose of about 300 mg of eculizumab, or an antigen-binding fragment thereof, thereby treating the subject, wherein the dsRNA agent comprises a sense strand and an antisense strand, wherein the sense strand comprises 5'-asasGfcAfaGfaUfAfUfuUfuuAfuAfaua-3' (SEQ ID NO: 2876), and the antisense strand comprises 5'-usAfsUfuAfuaAfaAfauaUfcUfuGfcuususudTdT-3' (SEQ ID NO: 2889), wherein a, g, c, and u are 2'-O-methyl (2'-OMe) A, G, C, and U, respectively; Af, Gf, Cf, and Uf are 2'-fluoro A, G, C, and U, respectively; dT is a deoxy-thymine nucleotide; and s is a phosphorothioate linkage. In one embodiment, the dsRNA agent is administered to the subject once a month, at a fixed dose of 200 mg, for 2 months, e.g., prior to administration of a dose of about 300 mg of eculizumab, or an antigen-binding fragment thereof, to the subject. In one embodiment, the dsRNA agent is administered to the subject once a month, at a fixed dose of 200 mg, for 2 months, e.g., prior to administration of a dose of about 300 mg of eculizumab, or an antigen-binding fragment thereof, to the subject, and then once every 3 months. In one embodiment, the dsRNA agent is administered to the subject once a month, at a fixed dose of 200 mg, for 2 months, e.g., prior to administration of a dose of about 300 mg of eculizumab, or an antigen-binding fragment thereof, to the subject, and then once every 6 months. In another embodiment, the dsRNA agent is administered to the subject once a month, at a fixed dose of 200 mg, for 3 months, e.g., prior to administration of a dose of about 300 mg of eculizumab, or an antigen-binding fragment thereof, to the subject.In one embodiment, the dsRNA agent is administered to the subject, for example, at a fixed dose of 200 mg of the dsRNA agent once a month for 3 months, and then once every 3 months, prior to administration of about 300 mg of eculizumab, or an antigen-binding fragment thereof, to the subject. In one embodiment, the dsRNA agent is administered to the subject, for example, at a fixed dose of 200 mg of the dsRNA agent once a month for 3 months, and then once every 6 months, prior to administration of about 300 mg of eculizumab, or an antigen-binding fragment thereof, to the subject. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 200 mg of the dsRNA agent once a month for a long period of time.

[0048] In another aspect, the present invention provides a method for treating, e.g., treating over a long period of time, a subject suffering from paroxysmal nocturnal hemoglobinuria (PNH). The method comprises administering to a subject suffering from PNH and previously treated with eculizumab, once a month, a fixed dose of 200 mg of a double-stranded ribonucleic acid (dsRNA) agent for inhibiting the expression of complement component C5; and administering to the subject, once every 4 weeks, a dose of about 600 mg of eculizumab, or an antigen-binding fragment thereof, thereby treating the subject, wherein the dsRNA agent comprises a sense strand and an antisense strand, wherein the sense strand comprises 5’-asasGfcAfaGfaUfAfUfuUfuuAfuAfaua-3’ (SEQ ID NO: 2876), and the antisense strand comprises 5’-usAfsUfuAfuaAfaAfauaUfcUfuGfcuususudTdT-3’ (SEQ ID NO: 2889), wherein a, g, c, and u are 2’-O-methyl (2’-OMe) A, G, C, and U, respectively; Af, Gf, Cf, and Uf are 2’-fluoro A, G, C, and U, respectively; dT is deoxy-thymidine nucleotide; and s is a phosphorothioate bond. In one embodiment, the dsRNA agent is administered to the subject once a month for 2 months at a fixed dose of 200 mg of the dsRNA agent, e.g., prior to administration of a dose of about 600 mg of eculizumab, or an antigen-binding fragment thereof, to the subject. In one embodiment, the dsRNA agent is administered to the subject once a month for 2 months at a fixed dose of 200 mg of the dsRNA agent, e.g., prior to administration of a dose of about 600 mg of eculizumab, or an antigen-binding fragment thereof, to the subject, and then once every 3 months. In one embodiment, the dsRNA agent is administered to the subject once a month for 2 months at a fixed dose of 200 mg of the dsRNA agent, e.g., prior to administration of a dose of about 600 mg of eculizumab, or an antigen-binding fragment thereof, to the subject, and then once every 6 months. In another embodiment, the dsRNA agent is administered to the subject once a month for 3 months at a fixed dose of 200 mg of the dsRNA agent, e.g., prior to administration of a dose of about 600 mg of eculizumab, or an antigen-binding fragment thereof, to the subject.In one embodiment, the dsRNA agent is administered to the subject, for example, at a fixed dose of 200 mg of the dsRNA agent once a month for 3 months prior to administration of about 600 mg of eculizumab, or an antigen-binding fragment thereof, to the subject, and then once every 3 months thereafter. In one embodiment, the dsRNA agent is administered to the subject, for example, at a fixed dose of 200 mg of the dsRNA agent once a month for 3 months prior to administration of about 600 mg of eculizumab, or an antigen-binding fragment thereof, to the subject, and then once every 6 months thereafter. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 200 mg of the dsRNA agent once a month for a long period of time.

[0049] In another aspect, the present invention provides a method for treating, e.g., treating over a long period of time, a subject suffering from paroxysmal nocturnal hemoglobinuria (PNH). The method comprises administering to a subject suffering from PNH and previously treated with eculizumab, once a month, a fixed dose of 200 mg of a double-stranded ribonucleic acid (dsRNA) agent for inhibiting the expression of complement component C5; and administering to the subject, once every 4 weeks, a dose of about 900 mg of eculizumab, or an antigen-binding fragment thereof, thereby treating the subject, wherein the dsRNA agent comprises a sense strand and an antisense strand, wherein the sense strand comprises 5'-asasGfcAfaGfaUfAfUfuUfuuAfuAfaua-3' (SEQ ID NO: 2876), and the antisense strand comprises 5'-usAfsUfuAfuaAfaAfauaUfcUfuGfcuususudTdT-3' (SEQ ID NO: 2889), wherein a, g, c, and u are 2'-O-methyl (2'-OMe) A, G, C, and U, respectively; Af, Gf, Cf, and Uf are 2'-fluoro A, G, C, and U, respectively; dT is a deoxy-thymine nucleotide; and s is a phosphorothioate bond. In one embodiment, the dsRNA agent is administered to the subject once a month, at a fixed dose of 200 mg, for 2 months, e.g., prior to administration of a dose of about 900 mg of eculizumab, or an antigen-binding fragment thereof, to the subject. In one embodiment, the dsRNA agent is administered to the subject once a month, at a fixed dose of 200 mg, for 2 months, e.g., prior to administration of a dose of about 900 mg of eculizumab, or an antigen-binding fragment thereof, to the subject, and then once every 3 months. In one embodiment, the dsRNA agent is administered to the subject once a month, at a fixed dose of 200 mg, for 2 months, e.g., prior to administration of a dose of about 900 mg of eculizumab, or an antigen-binding fragment thereof, to the subject, and then once every 6 months. In another embodiment, the dsRNA agent is administered to the subject once a month, at a fixed dose of 200 mg, for 3 months, e.g., prior to administration of a dose of about 900 mg of eculizumab, or an antigen-binding fragment thereof, to the subject.In one embodiment, the dsRNA agent is administered to the subject, for example, at a fixed dose of 200 mg of the dsRNA agent once a month for 3 months prior to administration of about 900 mg of eculizumab, or an antigen-binding fragment thereof, to the subject, and then once every 3 months thereafter. In one embodiment, the dsRNA agent is administered to the subject, for example, at a fixed dose of 200 mg of the dsRNA agent once a month for 3 months prior to administration of about 900 mg of eculizumab, or an antigen-binding fragment thereof, to the subject, and then once every 6 months thereafter. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 200 mg of the dsRNA agent once a month for a long period of time.

[0050] In one aspect, the present invention provides a method for treating, e.g., treating over a long period of time, a subject suffering from paroxysmal nocturnal hemoglobinuria (PNH). The method comprises administering to a subject suffering from PNH and who has not responded to treatment with eculizumab, a double-stranded ribonucleic acid (dsRNA) agent at a fixed dose of 200 mg once a week for inhibiting the expression of complement component C5; and administering to the subject eculizumab or an antigen-binding fragment thereof at a dose of about 300 mg to about 900 mg once every four weeks, thereby treating the subject, wherein the dsRNA agent comprises a sense strand and an antisense strand, wherein the sense strand comprises 5’-asasGfcAfaGfaUfAfUfuUfuuAfuAfaua-3’ (SEQ ID NO: 2876), and the antisense strand comprises 5’-usAfsUfuAfuaAfaAfauaUfcUfuGfcuususudTdT-3’ (SEQ ID NO: 2889), wherein a, g, c, and u are 2’-O-methyl (2’-OMe) A, G, C, and U, respectively; Af, Gf, Cf, and Uf are 2’-fluoro A, G, C, and U, respectively; dT is a deoxy-thymine nucleotide; and s is a phosphorothioate bond. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 200 mg once a week for 8 weeks, e.g., prior to administration of eculizumab or an antigen-binding fragment thereof to the subject at a dose of about 300 mg to about 900 mg. In another embodiment, the dsRNA agent is administered to the subject at a fixed dose of 200 mg once a week for 12 weeks, e.g., prior to administration of eculizumab or an antigen-binding fragment thereof to the subject at a dose of about 300 mg to about 900 mg. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 200 mg once a week for 8 weeks, e.g., prior to administration of eculizumab or an antigen-binding fragment thereof to the subject at a dose of about 300 mg to about 900 mg, and then once a month thereafter. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 200 mg once a week for 8 weeks, e.g., prior to administration of eculizumab or an antigen-binding fragment thereof to the subject at a dose of about 300 mg to about 900 mg, and then once every three months thereafter.In one embodiment, the dsRNA agent is administered to a subject once a week for 8 weeks at a fixed dose of 200 mg of the dsRNA agent, and then once every 6 months, prior to administration of eculizumab, for example, at a dose of about 300 mg to about 900 mg, or an antigen-binding fragment thereof. In one embodiment, the dsRNA agent is administered to a subject once a week for 12 weeks at a fixed dose of 200 mg of the dsRNA agent, and then once a month, prior to administration of eculizumab, for example, at a dose of about 300 mg to about 900 mg, or an antigen-binding fragment thereof. In one embodiment, the dsRNA agent is administered to a subject once a week for 12 weeks at a fixed dose of 200 mg of the dsRNA agent, and then once every 3 months, prior to administration of eculizumab, for example, at a dose of about 300 mg to about 900 mg, or an antigen-binding fragment thereof. In one embodiment, the dsRNA agent is administered to a subject once a week for 12 weeks at a fixed dose of 200 mg of the dsRNA agent, and then once every 6 months, prior to administration of eculizumab, for example, at a dose of about 300 mg to about 900 mg, or an antigen-binding fragment thereof. In one embodiment, the dsRNA agent is administered to a subject at a fixed dose of 200 mg of the dsRNA agent once a week for a long period of time.

[0051] In one aspect, the present invention provides a method for treating, e.g., treating over a long period of time, a subject suffering from paroxysmal nocturnal hemoglobinuria (PNH). The method comprises administering to a subject suffering from PNH and who has not responded to treatment with eculizumab a double-stranded ribonucleic acid (dsRNA) agent at a fixed dose of 200 mg once a week for inhibiting the expression of complement component C5; and administering to the subject eculizumab or an antigen-binding fragment thereof at a dose of about 300 mg once every four weeks, thereby treating the subject, wherein the dsRNA agent comprises a sense strand and an antisense strand, wherein the sense strand comprises 5’-asasGfcAfaGfaUfAfUfuUfuuAfuAfaua-3’ (SEQ ID NO: 2876), and the antisense strand comprises 5’-usAfsUfuAfuaAfaAfauaUfcUfuGfcuususudTdT-3’ (SEQ ID NO: 2889), wherein a, g, c, and u are 2’-O-methyl (2’-OMe) A, G, C, and U, respectively; Af, Gf, Cf, and Uf are 2’-fluoro A, G, C, and U, respectively; dT is a deoxy-thymine nucleotide; and s is a phosphorothioate bond. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 200 mg once a week for 8 weeks, e.g., prior to administration of eculizumab or an antigen-binding fragment thereof at a dose of about 300 mg to the subject. In another embodiment, the dsRNA agent is administered to the subject at a fixed dose of 200 mg once a week for 12 weeks, e.g., prior to administration of eculizumab or an antigen-binding fragment thereof at a dose of about 300 mg to the subject. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 200 mg once a week for 8 weeks, e.g., prior to administration of eculizumab or an antigen-binding fragment thereof at a dose of about 300 mg to the subject, and then once a month thereafter. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 200 mg once a week for 8 weeks, e.g., prior to administration of eculizumab or an antigen-binding fragment thereof at a dose of about 300 mg to the subject, and then once every three months thereafter.In one embodiment, the dsRNA agent is administered to the subject once a week for 8 weeks at a fixed dose of 200 mg of the dsRNA agent, and then once every 6 months, prior to administration of, for example, a dose of about 300 mg of eculizumab or an antigen-binding fragment thereof. In one embodiment, the dsRNA agent is administered to the subject once a week for 12 weeks at a fixed dose of 200 mg of the dsRNA agent, and then once a month, prior to administration of, for example, a dose of about 300 mg of eculizumab or an antigen-binding fragment thereof. In one embodiment, the dsRNA agent is administered to the subject once a week for 12 weeks at a fixed dose of 200 mg of the dsRNA agent, and then once every 3 months, prior to administration of, for example, a dose of about 300 mg of eculizumab or an antigen-binding fragment thereof. In one embodiment, the dsRNA agent is administered to the subject once a week for 12 weeks at a fixed dose of 200 mg of the dsRNA agent, and then once every 6 months, prior to administration of, for example, a dose of about 300 mg of eculizumab or an antigen-binding fragment thereof. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 200 mg of the dsRNA agent once a week for a long period of time.

[0052] In one aspect, the present invention provides a method for treating, e.g., treating over a long period of time, a subject suffering from paroxysmal nocturnal hemoglobinuria (PNH). The method comprises administering to a subject suffering from PNH and not responding to treatment with eculizumab a double-stranded ribonucleic acid (dsRNA) agent at a fixed dose of 200 mg once a week for inhibiting the expression of complement component C5; and administering to the subject eculizumab or an antigen-binding fragment thereof at a dose of about 600 mg once every four weeks, thereby treating the subject, wherein the dsRNA agent comprises a sense strand and an antisense strand, wherein the sense strand comprises 5'-asasGfcAfaGfaUfAfUfuUfuuAfuAfaua-3' (SEQ ID NO: 2876), and the antisense strand comprises 5'-usAfsUfuAfuaAfaAfauaUfcUfuGfcuususudTdT-3' (SEQ ID NO: 2889), wherein a, g, c, and u are 2'-O-methyl (2'-OMe) A, G, C, and U, respectively; Af, Gf, Cf, and Uf are 2'-fluoro A, G, C, and U, respectively; dT is deoxy-thymidine nucleotide; and s is a phosphorothioate bond. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 200 mg once a week for 8 weeks, e.g., prior to administration of eculizumab or an antigen-binding fragment thereof at a dose of about 600 mg to the subject. In another embodiment, the dsRNA agent is administered to the subject at a fixed dose of 200 mg once a week for 12 weeks, e.g., prior to administration of eculizumab or an antigen-binding fragment thereof at a dose of about 600 mg to the subject. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 200 mg once a week for 8 weeks, e.g., prior to administration of eculizumab or an antigen-binding fragment thereof at a dose of about 600 mg to the subject, and then once a month thereafter. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 200 mg once a week for 8 weeks, e.g., prior to administration of eculizumab or an antigen-binding fragment thereof at a dose of about 600 mg to the subject, and then once every three months thereafter.In one embodiment, the dsRNA agent is administered to a subject once a week for 8 weeks at a fixed dose of 200 mg of the dsRNA agent, and then once every 6 months, prior to administration of, for example, a dose of about 600 mg of eculizumab, or an antigen-binding fragment thereof. In one embodiment, the dsRNA agent is administered to a subject once a week for 12 weeks at a fixed dose of 200 mg of the dsRNA agent, and then once a month, prior to administration of, for example, a dose of about 600 mg of eculizumab, or an antigen-binding fragment thereof. In one embodiment, the dsRNA agent is administered to a subject once a week for 12 weeks at a fixed dose of 200 mg of the dsRNA agent, and then once every 3 months, prior to administration of, for example, a dose of about 600 mg of eculizumab, or an antigen-binding fragment thereof. In one embodiment, the dsRNA agent is administered to a subject once a week for 12 weeks at a fixed dose of 200 mg of the dsRNA agent, and then once every 6 months, prior to administration of, for example, a dose of about 600 mg of eculizumab, or an antigen-binding fragment thereof. In one embodiment, the dsRNA agent is administered to a subject at a fixed dose of 200 mg of the dsRNA agent once a week for a long period of time.

[0053] In one aspect, the present invention provides a method for treating, e.g., treating over a long period of time, a subject suffering from paroxysmal nocturnal hemoglobinuria (PNH). The method comprises administering to a subject suffering from PNH and not responding to treatment with eculizumab a double-stranded ribonucleic acid (dsRNA) agent at a fixed dose of 200 mg once a week for inhibiting the expression of complement component C5; and administering to the subject eculizumab at a dose of about 900 mg once every four weeks, or an antigen-binding fragment thereof, thereby treating the subject, wherein the dsRNA agent comprises a sense strand and an antisense strand, wherein the sense strand comprises 5'-asasGfcAfaGfaUfAfUfuUfuuAfuAfaua-3' (SEQ ID NO: 2876), the antisense strand comprises 5'-usAfsUfuAfuaAfaAfauaUfcUfuGfcuususudTdT-3' (SEQ ID NO: 2889), wherein a, g, c, and u are 2'-O-methyl (2'-OMe) A, G, C, and U, respectively; Af, Gf, Cf, and Uf are 2'-fluoro A, G, C, and U, respectively; dT is deoxy-thymidine nucleotide; and s is a phosphorothioate bond. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 200 mg once a week for 8 weeks, e.g., prior to administration of eculizumab at a dose of about 900 mg, or an antigen-binding fragment thereof, to the subject. In another embodiment, the dsRNA agent is administered to the subject at a fixed dose of 200 mg once a week for 12 weeks, e.g., prior to administration of eculizumab at a dose of about 900 mg, or an antigen-binding fragment thereof, to the subject. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 200 mg once a week for 8 weeks, e.g., prior to administration of eculizumab at a dose of about 900 mg, or an antigen-binding fragment thereof, to the subject, and then once a month thereafter. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 200 mg once a week for 8 weeks, e.g., prior to administration of eculizumab at a dose of about 900 mg, or an antigen-binding fragment thereof, to the subject, and then once every three months thereafter.In one embodiment, the dsRNA agent is administered to the subject once a week for 8 weeks at a fixed dose of 200 mg of the dsRNA agent, and then once every 6 months, prior to administration of, for example, a dose of about 900 mg of eculizumab, or an antigen-binding fragment thereof. In one embodiment, the dsRNA agent is administered to the subject once a week for 12 weeks at a fixed dose of 200 mg of the dsRNA agent, and then once a month, prior to administration of, for example, a dose of about 900 mg of eculizumab, or an antigen-binding fragment thereof. In one embodiment, the dsRNA agent is administered to the subject once a week for 12 weeks at a fixed dose of 200 mg of the dsRNA agent, and then once every 3 months, prior to administration of, for example, a dose of about 900 mg of eculizumab, or an antigen-binding fragment thereof. In one embodiment, the dsRNA agent is administered to the subject once a week for 12 weeks at a fixed dose of 200 mg of the dsRNA agent, and then once every 6 months, prior to administration of, for example, a dose of about 900 mg of eculizumab, or an antigen-binding fragment thereof. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 200 mg once a week for a long period of time.

[0054] In another aspect, the present invention provides a method for treating, e.g., treating over a long period of time, a subject suffering from paroxysmal nocturnal hemoglobinuria (PNH). The method comprises administering to a subject suffering from PNH and who has not responded to treatment with eculizumab, once a month, a fixed dose of 200 mg of a double-stranded ribonucleic acid (dsRNA) agent for inhibiting the expression of complement component C5; and administering to the subject, once every 4 weeks, a dose of eculizumab, or an antigen-binding fragment thereof, of about 300 mg to about 900 mg, thereby treating the subject, wherein the dsRNA agent comprises a sense strand and an antisense strand, wherein the sense strand comprises 5’-asasGfcAfaGfaUfAfUfuUfuuAfuAfaua-3’ (SEQ ID NO: 2876), and the antisense strand comprises 5’-usAfsUfuAfuaAfaAfauaUfcUfuGfcuususudTdT-3’ (SEQ ID NO: 2889), wherein a, g, c, and u are 2’-O-methyl (2’-OMe) A, G, C, and U, respectively; Af, Gf, Cf, and Uf are 2’-fluoro A, G, C, and U, respectively; dT is a deoxy-thymine nucleotide; and s is a phosphorothioate bond. In one embodiment, the dsRNA agent is administered to the subject once a month, at a fixed dose of 200 mg, for 2 months, e.g., prior to administration of a dose of eculizumab, or an antigen-binding fragment thereof, of about 300 mg to about 900 mg to the subject. In one embodiment, the dsRNA agent is administered to the subject once a month, at a fixed dose of 200 mg, for 2 months, e.g., prior to administration of a dose of eculizumab, or an antigen-binding fragment thereof, of about 300 mg to about 900 mg to the subject, and then once every 3 months. In one embodiment, the dsRNA agent is administered to the subject once a month, at a fixed dose of 200 mg, for 2 months, e.g., prior to administration of a dose of eculizumab, or an antigen-binding fragment thereof, of about 300 mg to about 900 mg to the subject, and then once every 6 months. In another embodiment, the dsRNA agent is administered to the subject once a month, at a fixed dose of 200 mg, for 3 months, e.g., prior to administration of a dose of eculizumab, or an antigen-binding fragment thereof, of about 300 mg to about 900 mg to the subject.In one embodiment, the dsRNA agent is administered to the subject, for example, at a fixed dose of 200 mg of the dsRNA agent once a month for 3 months, and then once every 3 months, prior to administration of eculizumab, or an antigen-binding fragment thereof, at a dose of about 300 mg to about 900 mg. In one embodiment, the dsRNA agent is administered to the subject, for example, at a fixed dose of 200 mg of the dsRNA agent once a month for 3 months, and then once every 6 months, prior to administration of eculizumab, or an antigen-binding fragment thereof, at a dose of about 300 mg to about 900 mg. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 200 mg of the dsRNA agent once a month for a long period of time.

[0055] In another aspect, the present invention provides a method for treating, e.g., treating over a long period of time, a subject suffering from paroxysmal nocturnal hemoglobinuria (PNH). The method comprises administering to a subject suffering from PNH and who has not responded to treatment with eculizumab, once a month, a fixed dose of 200 mg of a double-stranded ribonucleic acid (dsRNA) agent for inhibiting the expression of complement component C5; and administering to the subject, once every 4 weeks, a dose of about 300 mg of eculizumab, or an antigen-binding fragment thereof, thereby treating the subject, wherein the dsRNA agent comprises a sense strand and an antisense strand, wherein the sense strand comprises 5’-asasGfcAfaGfaUfAfUfuUfuuAfuAfaua-3’ (SEQ ID NO: 2876), and the antisense strand comprises 5’-usAfsUfuAfuaAfaAfauaUfcUfuGfcuususudTdT-3’ (SEQ ID NO: 2889), wherein a, g, c, and u are 2’-O-methyl (2’-OMe) A, G, C, and U, respectively; Af, Gf, Cf, and Uf are 2’-fluoro A, G, C, and U, respectively; dT is deoxy-thymidine nucleotide; and s is a phosphorothioate bond. In one embodiment, the dsRNA agent is administered to the subject once a month, at a fixed dose of 200 mg, for 2 months, e.g., prior to administration of a dose of about 300 mg of eculizumab, or an antigen-binding fragment thereof, to the subject. In one embodiment, the dsRNA agent is administered to the subject once a month, at a fixed dose of 200 mg, for 2 months, e.g., prior to administration of a dose of about 300 mg of eculizumab, or an antigen-binding fragment thereof, to the subject, and then once every 3 months thereafter. In one embodiment, the dsRNA agent is administered to the subject once a month, at a fixed dose of 200 mg, for 2 months, e.g., prior to administration of a dose of about 300 mg of eculizumab, or an antigen-binding fragment thereof, to the subject, and then once every 6 months thereafter. In another embodiment, the dsRNA agent is administered to the subject once a month, at a fixed dose of 200 mg, for 3 months, e.g., prior to administration of a dose of about 300 mg of eculizumab, or an antigen-binding fragment thereof, to the subject.In one embodiment, the dsRNA agent is administered to the subject, for example, at a fixed dose of 200 mg of the dsRNA agent once a month for 3 months, and then once every 3 months, prior to administration of a dose of about 300 mg of eculizumab, or an antigen-binding fragment thereof, to the subject. In one embodiment, the dsRNA agent is administered to the subject, for example, at a fixed dose of 200 mg of the dsRNA agent once a month for 3 months, and then once every 6 months, prior to administration of a dose of about 300 mg of eculizumab, or an antigen-binding fragment thereof, to the subject. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 200 mg of the dsRNA agent once a month for a long period of time.

[0056] In another aspect, the present invention provides a method for treating, e.g., treating over a long period of time, a subject suffering from paroxysmal nocturnal hemoglobinuria (PNH). The method comprises administering to a subject suffering from PNH and who has not responded to treatment with eculizumab a double-stranded ribonucleic acid (dsRNA) agent at a fixed dose of 200 mg once a month for inhibiting the expression of complement component C5; and administering to the subject eculizumab or an antigen-binding fragment thereof at a dose of about 600 mg once every 4 weeks, thereby treating the subject, wherein the dsRNA agent comprises a sense strand and an antisense strand, wherein the sense strand comprises 5'-asasGfcAfaGfaUfAfUfuUfuuAfuAfaua-3' (SEQ ID NO: 2876), and the antisense strand comprises 5'-usAfsUfuAfuaAfaAfauaUfcUfuGfcuususudTdT-3' (SEQ ID NO: 2889), wherein a, g, c, and u are 2'-O-methyl (2'-OMe) A, G, C, and U, respectively; Af, Gf, Cf, and Uf are 2'-fluoro A, G, C, and U, respectively; dT is a deoxy-thymine nucleotide; and s is a phosphorothioate bond. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 200 mg once a month for 2 months, e.g., prior to administration of eculizumab or an antigen-binding fragment thereof at a dose of about 600 mg to the subject. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 200 mg once a month for 2 months, e.g., prior to administration of eculizumab or an antigen-binding fragment thereof at a dose of about 600 mg to the subject, and then once every 3 months thereafter. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 200 mg once a month for 2 months, e.g., prior to administration of eculizumab or an antigen-binding fragment thereof at a dose of about 600 mg to the subject, and then once every 6 months thereafter. In another embodiment, the dsRNA agent is administered to the subject at a fixed dose of 200 mg once a month for 3 months, e.g., prior to administration of eculizumab or an antigen-binding fragment thereof at a dose of about 600 mg to the subject.In one embodiment, the dsRNA agent is administered to the subject, for example, at a fixed dose of 200 mg of the dsRNA agent once a month for 3 months, and then once every 3 months, prior to administration of a dose of about 600 mg of eculizumab, or an antigen-binding fragment thereof, to the subject. In one embodiment, the dsRNA agent is administered to the subject, for example, at a fixed dose of 200 mg of the dsRNA agent once a month for 3 months, and then once every 6 months, prior to administration of a dose of about 600 mg of eculizumab, or an antigen-binding fragment thereof, to the subject. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 200 mg of the dsRNA agent once a month for a long period of time.

[0057] In another aspect, the present invention provides a method for treating, e.g., treating over a long period of time, a subject suffering from paroxysmal nocturnal hemoglobinuria (PNH). The method comprises administering to a subject suffering from PNH and not responsive to treatment with eculizumab, once a month, a fixed dose of 200 mg of a double-stranded ribonucleic acid (dsRNA) agent for inhibiting the expression of complement component C5; and administering to the subject, once every 4 weeks, a dose of about 900 mg of eculizumab, or an antigen-binding fragment thereof, thereby treating the subject, wherein the dsRNA agent comprises a sense strand and an antisense strand, wherein the sense strand comprises 5’-asasGfcAfaGfaUfAfUfuUfuuAfuAfaua-3’ (SEQ ID NO: 2876), and the antisense strand comprises 5’-usAfsUfuAfuaAfaAfauaUfcUfuGfcuususudTdT-3’ (SEQ ID NO: 2889), wherein a, g, c, and u are 2’-O-methyl (2’-OMe) A, G, C, and U, respectively; Af, Gf, Cf, and Uf are 2’-fluoro A, G, C, and U, respectively; dT is deoxy-thymidine nucleotide; and s is a phosphorothioate linkage. In one embodiment, the dsRNA agent is administered to the subject once a month at a fixed dose of 200 mg for 2 months, e.g., prior to administration of a dose of about 900 mg of eculizumab, or an antigen-binding fragment thereof, to the subject. In one embodiment, the dsRNA agent is administered to the subject once a month at a fixed dose of 200 mg for 2 months, e.g., prior to administration of a dose of about 900 mg of eculizumab, or an antigen-binding fragment thereof, to the subject, and then once every 3 months. In one embodiment, the dsRNA agent is administered to the subject once a month at a fixed dose of 200 mg for 2 months, e.g., prior to administration of a dose of about 900 mg of eculizumab, or an antigen-binding fragment thereof, to the subject, and then once every 6 months. In another embodiment, the dsRNA agent is administered to the subject once a month at a fixed dose of 200 mg for 3 months, e.g., prior to administration of a dose of about 900 mg of eculizumab, or an antigen-binding fragment thereof, to the subject.In one embodiment, the dsRNA agent is administered to a subject, for example, once a month for 3 months at a fixed dose of 200 mg of the dsRNA agent, and then once every 3 months, prior to administration of about 900 mg of eculizumab, or an antigen-binding fragment thereof, to the subject. In one embodiment, the dsRNA agent is administered to a subject, for example, once a month for 3 months at a fixed dose of 200 mg of the dsRNA agent, and then once every 6 months, prior to administration of about 900 mg of eculizumab, or an antigen-binding fragment thereof, to the subject. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 200 mg once a month for a long period of time. In one aspect, the present invention provides a method for treating, for example, treating for a long period of time, a subject suffering from paroxysmal nocturnal hemoglobinuria (PNH). The method comprises administering to a subject suffering from PNH and not responding to treatment with eculizumab a double-stranded ribonucleic acid (dsRNA) agent at a fixed dose of 400 mg once a week to inhibit the expression of complement component C5; and administering to the subject eculizumab, or an antigen-binding fragment thereof, at a dose of about 300 mg to about 900 mg once every 4 weeks, thereby treating the subject, wherein the dsRNA agent comprises a sense strand and an antisense strand, wherein the sense strand comprises 5'-asasGfcAfaGfaUfAfUfuUfuuAfuAfaua-3' (SEQ ID NO: 2876), the antisense strand comprises 5'-usAfsUfuAfuaAfaAfauaUfcUfuGfcuususudTdT-3' (SEQ ID NO: 2889), wherein a, g, c, and u are 2'-O-methyl (2'-OMe) A, G, C, and U, respectively; Af, Gf, Cf, and Uf are 2'-fluoro A, G, C, and U, respectively; dT is a deoxy-thymine nucleotide; and s is a phosphorothioate bond. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 400 mg once a week for 8 weeks prior to administration of about 300 mg to about 900 mg of eculizumab, or an antigen-binding fragment thereof, to the subject.In another embodiment, the dsRNA agent is administered to the subject once a week for 12 weeks at a fixed dose of 400 mg of the dsRNA agent, for example, prior to administration of a dose of eculizumab of about 300 mg to about 900 mg, or an antigen-binding fragment thereof. In one embodiment, the dsRNA agent is administered to the subject once a week for 8 weeks at a fixed dose of 400 mg of the dsRNA agent, for example, prior to administration of a dose of eculizumab of about 300 mg to about 900 mg, or an antigen-binding fragment thereof, and then once a month thereafter. In one embodiment, the dsRNA agent is administered to the subject once a week for 8 weeks at a fixed dose of 400 mg of the dsRNA agent, for example, prior to administration of a dose of eculizumab of about 300 mg to about 900 mg, or an antigen-binding fragment thereof, and then once every 3 months thereafter. In one embodiment, the dsRNA agent is administered to the subject once a week for 8 weeks at a fixed dose of 400 mg of the dsRNA agent, for example, prior to administration of a dose of eculizumab of about 300 mg to about 900 mg, or an antigen-binding fragment thereof, and then once every 6 months thereafter. In one embodiment, the dsRNA agent is administered to the subject once a week for 12 weeks at a fixed dose of 400 mg of the dsRNA agent, for example, prior to administration of a dose of eculizumab of about 300 mg to about 900 mg, or an antigen-binding fragment thereof, and then once a month thereafter. In one embodiment, the dsRNA agent is administered to the subject once a week for 12 weeks at a fixed dose of 400 mg of the dsRNA agent, for example, prior to administration of a dose of eculizumab of about 300 mg to about 900 mg, or an antigen-binding fragment thereof, and then once every 3 months thereafter. In one embodiment, the dsRNA agent is administered to the subject once a week for 12 weeks at a fixed dose of 400 mg of the dsRNA agent, for example, prior to administration of a dose of eculizumab of about 300 mg to about 900 mg, or an antigen-binding fragment thereof, and then once every 6 months thereafter. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 400 mg of the dsRNA agent once a week for a long period of time.

[0058] In one aspect, the present invention provides a method for treating, e.g., treating over a long period of time, a subject suffering from paroxysmal nocturnal hemoglobinuria (PNH). The method comprises administering to a subject suffering from PNH and not responsive to treatment with eculizumab a double-stranded ribonucleic acid (dsRNA) agent at a fixed dose of 400 mg once a week for inhibiting the expression of complement component C5; and administering to the subject eculizumab or an antigen-binding fragment thereof at a dose of about 300 mg once every 4 weeks, thereby treating the subject, wherein the dsRNA agent comprises a sense strand and an antisense strand, wherein the sense strand comprises 5'-asasGfcAfaGfaUfAfUfuUfuuAfuAfaua-3' (SEQ ID NO: 2876), and the antisense strand comprises 5'-usAfsUfuAfuaAfaAfauaUfcUfuGfcuususudTdT-3' (SEQ ID NO: 2889), wherein a, g, c, and u are 2'-O-methyl (2'-OMe) A, G, C, and U, respectively; Af, Gf, Cf, and Uf are 2'-fluoro A, G, C, and U, respectively; dT is deoxy-thymidine nucleotide; and s is a phosphorothioate bond. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 400 mg once a week for 8 weeks, e.g., prior to administration of eculizumab or an antigen-binding fragment thereof at a dose of about 300 mg to the subject. In another embodiment, the dsRNA agent is administered to the subject at a fixed dose of 400 mg once a week for 12 weeks, e.g., prior to administration of eculizumab or an antigen-binding fragment thereof at a dose of about 300 mg to the subject. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 400 mg once a week for 8 weeks, e.g., prior to administration of eculizumab or an antigen-binding fragment thereof at a dose of about 300 mg to the subject, and then once a month thereafter. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 400 mg once a week for 8 weeks, e.g., prior to administration of eculizumab or an antigen-binding fragment thereof at a dose of about 300 mg to the subject, and then once every 3 months thereafter.In one embodiment, the dsRNA agent is administered to the subject once a week for 8 weeks at a fixed dose of 400 mg of the dsRNA agent, and then once every 6 months, for example, before administration of a dose of about 300 mg of eculizumab, or an antigen-binding fragment thereof. In one embodiment, the dsRNA agent is administered to the subject once a week for 12 weeks at a fixed dose of 400 mg of the dsRNA agent, and then once a month, for example, before administration of a dose of about 300 mg of eculizumab, or an antigen-binding fragment thereof. In one embodiment, the dsRNA agent is administered to the subject once a week for 12 weeks at a fixed dose of 400 mg of the dsRNA agent, and then once every 3 months, for example, before administration of a dose of about 300 mg of eculizumab, or an antigen-binding fragment thereof. In one embodiment, the dsRNA agent is administered to the subject once a week for 12 weeks at a fixed dose of 400 mg of the dsRNA agent, and then once every 6 months, for example, before administration of a dose of about 300 mg of eculizumab, or an antigen-binding fragment thereof. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 400 mg of the dsRNA agent once a week for a long period of time.

[0059] In one aspect, the present invention provides a method for treating, e.g., treating over a long period of time, a subject suffering from paroxysmal nocturnal hemoglobinuria (PNH). The method comprises administering to a subject suffering from PNH and who has not responded to treatment with eculizumab a double-stranded ribonucleic acid (dsRNA) agent at a fixed dose of 400 mg once a week for inhibiting the expression of complement component C5; and administering to the subject eculizumab or an antigen-binding fragment thereof at a dose of about 600 mg once every four weeks, thereby treating the subject, wherein the dsRNA agent comprises a sense strand and an antisense strand, wherein the sense strand comprises 5'-asasGfcAfaGfaUfAfUfuUfuuAfuAfaua-3' (SEQ ID NO: 2876), and the antisense strand comprises 5'-usAfsUfuAfuaAfaAfauaUfcUfuGfcuususudTdT-3' (SEQ ID NO: 2889), wherein a, g, c, and u are 2'-O-methyl (2'-OMe) A, G, C, and U, respectively; Af, Gf, Cf, and Uf are 2'-fluoro A, G, C, and U, respectively; dT is a deoxy-thymine nucleotide; and s is a phosphorothioate bond. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 400 mg once a week for 8 weeks, e.g., prior to administration of eculizumab or an antigen-binding fragment thereof at a dose of about 600 mg to the subject. In another embodiment, the dsRNA agent is administered to the subject at a fixed dose of 400 mg once a week for 12 weeks, e.g., prior to administration of eculizumab or an antigen-binding fragment thereof at a dose of about 600 mg to the subject. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 400 mg once a week for 8 weeks, e.g., prior to administration of eculizumab or an antigen-binding fragment thereof at a dose of about 600 mg to the subject, and then once a month thereafter. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 400 mg once a week for 8 weeks, e.g., prior to administration of eculizumab or an antigen-binding fragment thereof at a dose of about 600 mg to the subject, and then once every three months thereafter.In one embodiment, the dsRNA agent is administered to a subject once a week for 8 weeks at a fixed dose of 400 mg of the dsRNA agent, and then once every 6 months, for example, before administration of about 600 mg of eculizumab or an antigen-binding fragment thereof. In one embodiment, the dsRNA agent is administered to a subject once a week for 12 weeks at a fixed dose of 400 mg of the dsRNA agent, and then once a month, for example, before administration of about 600 mg of eculizumab or an antigen-binding fragment thereof. In one embodiment, the dsRNA agent is administered to a subject once a week for 12 weeks at a fixed dose of 400 mg of the dsRNA agent, and then once every 3 months, for example, before administration of about 600 mg of eculizumab or an antigen-binding fragment thereof. In one embodiment, the dsRNA agent is administered to a subject once a week for 12 weeks at a fixed dose of 400 mg of the dsRNA agent, and then once every 6 months, for example, before administration of about 600 mg of eculizumab or an antigen-binding fragment thereof. In one embodiment, the dsRNA agent is administered to a subject at a fixed dose of 400 mg of the dsRNA agent once a week for a long period of time.

[0060] In one aspect, the present invention provides a method for treating, e.g., treating over a long period of time, a subject suffering from paroxysmal nocturnal hemoglobinuria (PNH). The method comprises administering to a subject suffering from PNH and not responding to treatment with eculizumab a double-stranded ribonucleic acid (dsRNA) agent at a fixed dose of 400 mg once a week for inhibiting the expression of complement component C5; and administering to the subject eculizumab or an antigen-binding fragment thereof at a dose of about 900 mg once every four weeks, thereby treating the subject, wherein the dsRNA agent comprises a sense strand and an antisense strand, wherein the sense strand comprises 5'-asasGfcAfaGfaUfAfUfuUfuuAfuAfaua-3' (SEQ ID NO: 2876), and the antisense strand comprises 5'-usAfsUfuAfuaAfaAfauaUfcUfuGfcuususudTdT-3' (SEQ ID NO: 2889), wherein a, g, c, and u are 2'-O-methyl (2'-OMe) A, G, C, and U, respectively; Af, Gf, Cf, and Uf are 2'-fluoro A, G, C, and U, respectively; dT is deoxy-thymine nucleotide; and s is a phosphorothioate bond. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 400 mg once a week for 8 weeks, e.g., prior to administration of eculizumab or an antigen-binding fragment thereof at a dose of about 900 mg to the subject. In another embodiment, the dsRNA agent is administered to the subject at a fixed dose of 400 mg once a week for 12 weeks, e.g., prior to administration of eculizumab or an antigen-binding fragment thereof at a dose of about 900 mg to the subject. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 400 mg once a week for 8 weeks, e.g., prior to administration of eculizumab or an antigen-binding fragment thereof at a dose of about 900 mg to the subject, and then once a month thereafter. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 400 mg once a week for 8 weeks, e.g., prior to administration of eculizumab or an antigen-binding fragment thereof at a dose of about 900 mg to the subject, and then once every three months thereafter.In one embodiment, the dsRNA agent is administered to the subject once a week for 8 weeks at a fixed dose of 400 mg of the dsRNA agent, and then once every 6 months, for example, prior to administration of a dose of about 900 mg of eculizumab, or an antigen-binding fragment thereof. In one embodiment, the dsRNA agent is administered to the subject once a week for 12 weeks at a fixed dose of 400 mg of the dsRNA agent, and then once a month, for example, prior to administration of a dose of about 900 mg of eculizumab, or an antigen-binding fragment thereof. In one embodiment, the dsRNA agent is administered to the subject once a week for 12 weeks at a fixed dose of 400 mg of the dsRNA agent, and then once every 3 months, for example, prior to administration of a dose of about 900 mg of eculizumab, or an antigen-binding fragment thereof. In one embodiment, the dsRNA agent is administered to the subject once a week for 12 weeks at a fixed dose of 400 mg of the dsRNA agent, and then once every 6 months, for example, prior to administration of a dose of about 900 mg of eculizumab, or an antigen-binding fragment thereof. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 400 mg of the dsRNA agent once a week for a long period of time.

[0061] In another aspect, the present invention provides a method for treating, e.g., treating for a long period of time, a subject suffering from paroxysmal nocturnal hemoglobinuria (PNH). The method comprises administering to a subject suffering from PNH and not responsive to treatment with eculizumab a double-stranded ribonucleic acid (dsRNA) agent at a fixed dose of 400 mg once a month for inhibiting the expression of complement component C5; and administering to the subject eculizumab or an antigen-binding fragment thereof at a dose of about 300 mg to about 900 mg once every 4 weeks, thereby treating the subject, wherein the dsRNA agent comprises a sense strand and an antisense strand, wherein the sense strand comprises 5'-asasGfcAfaGfaUfAfUfuUfuuAfuAfaua-3' (SEQ ID NO: 2876), the antisense strand comprises 5'-usAfsUfuAfuaAfaAfauaUfcUfuGfcuususudTdT-3' (SEQ ID NO: 2889), wherein a, g, c, and u are 2'-O-methyl (2'-OMe) A, G, C, and U, respectively; Af, Gf, Cf, and Uf are 2'-fluoro A, G, C, and U, respectively; dT is deoxy-thymidine nucleotide; and s is a phosphorothioate linkage. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 400 mg once a month for 2 months prior to, e.g., administration of eculizumab or an antigen-binding fragment thereof to the subject at a dose of about 300 mg to about 900 mg. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 400 mg once a month for 2 months prior to, e.g., administration of eculizumab or an antigen-binding fragment thereof to the subject at a dose of about 300 mg to about 900 mg and then once every 3 months thereafter. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 400 mg once a month for 2 months prior to, e.g., administration of eculizumab or an antigen-binding fragment thereof to the subject at a dose of about 300 mg to about 900 mg and then once every 6 months thereafter. In another embodiment, the dsRNA agent is administered to the subject at a fixed dose of 400 mg once a month for 3 months prior to, e.g., administration of eculizumab or an antigen-binding fragment thereof to the subject at a dose of about 300 mg to about 900 mg.In one embodiment, the dsRNA agent is administered to the subject once a month for 3 months at a fixed dose of 400 mg of the dsRNA agent, and then once every 3 months, prior to administration of eculizumab at a dose of about 300 mg to about 900 mg, or an antigen-binding fragment thereof, to the subject. In one embodiment, the dsRNA agent is administered to the subject once a month for 3 months at a fixed dose of 400 mg of the dsRNA agent, and then once every 6 months, prior to administration of eculizumab at a dose of about 300 mg to about 900 mg, or an antigen-binding fragment thereof, to the subject. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 400 mg once a month for a long period of time. In another aspect, the present invention provides a method for treating, e.g., treating for a long period of time, a subject suffering from paroxysmal nocturnal hemoglobinuria (PNH). The method comprises administering to a subject suffering from PNH and not responsive to treatment with eculizumab a double-stranded ribonucleic acid (dsRNA) agent at a fixed dose of 400 mg once a month for inhibiting the expression of complement component C5; and administering to the subject eculizumab at a dose of about 300 mg or an antigen-binding fragment thereof once every 4 weeks, thereby treating the subject, wherein the dsRNA agent comprises a sense strand and an antisense strand, wherein the sense strand comprises 5'-asasGfcAfaGfaUfAfUfuUfuuAfuAfaua-3' (SEQ ID NO: 2876), the antisense strand comprises 5'-usAfsUfuAfuaAfaAfauaUfcUfuGfcuususudTdT-3' (SEQ ID NO: 2889), wherein a, g, c, and u are 2'-O-methyl (2'-OMe)A, G, C, and U, respectively; Af, Gf, Cf, and Uf are 2'-fluoro A, G, C, and U, respectively; dT is deoxy-thymine nucleotide; and s is a phosphorothioate bond. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 400 mg once a month for 2 months prior to administration of eculizumab at a dose of about 300 mg, or an antigen-binding fragment thereof, to the subject.In one embodiment, the dsRNA agent is administered to the subject, for example, at a fixed dose of 400 mg of the dsRNA agent once a month for 2 months, and then once every 3 months, prior to administration of about 300 mg of eculizumab, or an antigen-binding fragment thereof, to the subject. In one embodiment, the dsRNA agent is administered to the subject, for example, at a fixed dose of 400 mg of the dsRNA agent once a month for 2 months, and then once every 6 months, prior to administration of about 300 mg of eculizumab, or an antigen-binding fragment thereof, to the subject. In another embodiment, the dsRNA agent is administered to the subject, for example, at a fixed dose of 400 mg of the dsRNA agent once a month for 3 months, prior to administration of about 300 mg of eculizumab, or an antigen-binding fragment thereof, to the subject. In one embodiment, the dsRNA agent is administered to the subject, for example, at a fixed dose of 400 mg of the dsRNA agent once a month for 3 months, and then once every 3 months, prior to administration of about 300 mg of eculizumab, or an antigen-binding fragment thereof, to the subject. In one embodiment, the dsRNA agent is administered to the subject, for example, at a fixed dose of 400 mg of the dsRNA agent once a month for 3 months, and then once every 6 months, prior to administration of about 300 mg of eculizumab, or an antigen-binding fragment thereof, to the subject. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 400 mg of the dsRNA agent once a month for a long period of time.

[0062] In another aspect, the present invention provides a method for treating, e.g., treating over a long period of time, a subject suffering from paroxysmal nocturnal hemoglobinuria (PNH). The method comprises administering to a subject suffering from PNH and not responsive to treatment with eculizumab, once a month, a fixed dose of 400 mg of a double-stranded ribonucleic acid (dsRNA) agent for inhibiting the expression of complement component C5; and administering to the subject, once every 4 weeks, a dose of about 600 mg of eculizumab, or an antigen-binding fragment thereof, thereby treating the subject, wherein the dsRNA agent comprises a sense strand and an antisense strand, wherein the sense strand comprises 5’-asasGfcAfaGfaUfAfUfuUfuuAfuAfaua-3’ (SEQ ID NO: 2876), the antisense strand comprises 5’-usAfsUfuAfuaAfaAfauaUfcUfuGfcuususudTdT-3’ (SEQ ID NO: 2889), wherein a, g, c, and u are 2’-O-methyl (2’-OMe) A, G, C, and U, respectively; Af, Gf, Cf, and Uf are 2’-fluoro A, G, C, and U, respectively; dT is deoxy-thymidine nucleotide; and s is a phosphorothioate bond. In one embodiment, the dsRNA agent is administered to the subject once a month for 2 months at a fixed dose of 400 mg of the dsRNA agent, e.g., prior to administration of a dose of about 600 mg of eculizumab, or an antigen-binding fragment thereof, to the subject. In one embodiment, the dsRNA agent is administered to the subject once a month for 2 months at a fixed dose of 400 mg of the dsRNA agent, e.g., prior to administration of a dose of about 600 mg of eculizumab, or an antigen-binding fragment thereof, to the subject, and then once every 3 months. In one embodiment, the dsRNA agent is administered to the subject once a month for 2 months at a fixed dose of 400 mg of the dsRNA agent, e.g., prior to administration of a dose of about 600 mg of eculizumab, or an antigen-binding fragment thereof, to the subject, and then once every 6 months. In another embodiment, the dsRNA agent is administered to the subject once a month for 3 months at a fixed dose of 400 mg of the dsRNA agent, e.g., prior to administration of a dose of about 600 mg of eculizumab, or an antigen-binding fragment thereof, to the subject.In one embodiment, the dsRNA agent is administered to the subject, for example, at a fixed dose of 400 mg of the dsRNA agent once a month for 3 months, and then once every 3 months, prior to administration of about 600 mg of eculizumab, or an antigen-binding fragment thereof, to the subject. In one embodiment, the dsRNA agent is administered to the subject, for example, at a fixed dose of 400 mg of the dsRNA agent once a month for 3 months, and then once every 6 months, prior to administration of about 600 mg of eculizumab, or an antigen-binding fragment thereof, to the subject. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 400 mg of the dsRNA agent once a month for a long period of time.

[0063] In another aspect, the present invention provides a method for treating, e.g., treating over a long period of time, a subject suffering from paroxysmal nocturnal hemoglobinuria (PNH). The method comprises administering to a subject suffering from PNH and not responsive to treatment with eculizumab, once a month, a fixed dose of 400 mg of a double-stranded ribonucleic acid (dsRNA) agent for inhibiting the expression of complement component C5; and administering to the subject a dose of about 900 mg of eculizumab, or an antigen-binding fragment thereof, thereby treating the subject, wherein the dsRNA agent comprises a sense strand and an antisense strand, wherein the sense strand comprises 5’-asasGfcAfaGfaUfAfUfuUfuuAfuAfaua-3’ (SEQ ID NO: 2876), and the antisense strand comprises 5’-usAfsUfuAfuaAfaAfauaUfcUfuGfcuususudTdT-3’ (SEQ ID NO: 2889), wherein a, g, c, and u are 2’-O-methyl (2’-OMe) A, G, C, and U, respectively; Af, Gf, Cf, and Uf are 2’-fluoro A, G, C, and U, respectively; dT is a deoxy-thymine nucleotide; and s is a phosphorothioate bond. In one embodiment, the dsRNA agent is administered to the subject once a month for 2 months at a fixed dose of 400 mg of the dsRNA agent, e.g., prior to administration of a dose of about 900 mg of eculizumab, or an antigen-binding fragment thereof, to the subject. In one embodiment, the dsRNA agent is administered to the subject once a month for 2 months at a fixed dose of 400 mg of the dsRNA agent, e.g., prior to administration of a dose of about 900 mg of eculizumab, or an antigen-binding fragment thereof, to the subject, and then once every 3 months thereafter. In one embodiment, the dsRNA agent is administered to the subject once a month for 2 months at a fixed dose of 400 mg of the dsRNA agent, e.g., prior to administration of a dose of about 900 mg of eculizumab, or an antigen-binding fragment thereof, to the subject, and then once every 6 months thereafter. In another embodiment, the dsRNA agent is administered to the subject once a month for 3 months at a fixed dose of 400 mg of the dsRNA agent, e.g., prior to administration of a dose of about 900 mg of eculizumab, or an antigen-binding fragment thereof, to the subject.In one embodiment, the dsRNA agent is administered to the subject once a month for 3 months at a fixed dose of 400 mg of the dsRNA agent, and then once every 3 months, prior to administration of, for example, a dose of about 900 mg of eculizumab, or an antigen-binding fragment thereof, to the subject. In one embodiment, the dsRNA agent is administered to the subject once a month for 3 months at a fixed dose of 400 mg of the dsRNA agent, and then once every 6 months, prior to administration of, for example, a dose of about 900 mg of eculizumab, or an antigen-binding fragment thereof, to the subject. In one embodiment, the dsRNA agent is administered to the subject at a fixed dose of 400 mg of the dsRNA agent once a month for a long period of time.

[0064] In certain embodiments, the dose of eculizumab, or an antigen-binding fragment thereof, is about 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, or 75% of the eculizumab maintenance level dose. In one embodiment, the dose of eculizumab, or an antigen-binding fragment thereof, is about 25% - 75%, 25% - 70%, 25% - 65%, 25% - 60%, 30% - 75%, 30% - 70%, 30% - 65%, 30% - 60%, 25% - 50%, 25% - 40% or 25% - 30% of the eculizumab maintenance level dose.

[0065] In certain embodiments, the frequency of administration of eculizumab is decreased compared to the frequency of administration required by this level. In certain aspects, eculizumab is administered once every 4 weeks, once every 2 months, once every 3 months, once every 4 months, once every 5 months or once every 6 months.

[0066] In certain embodiments, eculizumab, or an antigen-binding fragment thereof, is administered to a subject as a fixed dose of 300 mg once monthly. In certain embodiments, eculizumab, or an antigen-binding fragment thereof, is administered to a subject as a fixed dose of 600 mg once monthly. In certain embodiments, eculizumab, or an antigen-binding fragment thereof, is administered to a subject as a fixed dose of 900 mg every other week. In other embodiments, eculizumab, or an antigen-binding fragment thereof, is administered to a subject as a fixed dose of 1200 mg over a four-week period, followed by a fixed dose of 900 mg every other week.

[0067] In certain embodiments, eculizumab, or an antigen-binding fragment thereof, is administered to a subject naïve to eculizumab as a fixed dose of 300 mg once every four weeks. In certain embodiments, eculizumab, or an antigen-binding fragment thereof, is administered to a subject naïve to eculizumab as a fixed dose of 600 mg once every four weeks. In other embodiments, eculizumab, or an antigen-binding fragment thereof, is administered to a subject previously treated with eculizumab as a fixed dose of 900 mg once every four weeks.

[0068] In one embodiment, previous treatment with eculizumab or an antigen-binding fragment thereof included administration of eculizumab at a fixed dose of 900 mg twice weekly to a subject. In one embodiment, eculizumab treatment to which a subject did not respond included administration of eculizumab at a fixed dose of 1200 mg over a four-week period to a subject.

[0069] In certain aspects, the dsRNA agent and eculizumab, or an antigen-binding fragment thereof, are administered to a subject simultaneously. In other aspects, the dsRNA agent is administered to a subject prior to eculizumab, or an antigen-binding fragment thereof. In yet other aspects, eculizumab, or an antigen-binding fragment thereof, is administered to a subject prior to the dsRNA agent.

[0070] In certain embodiments, the treatment prevents hemolytic attacks in a subject.

[0071] In certain embodiments, the treatment reduces the mean maximal C5 mRNA level by at least about 98%, for example, by at least about 99% relative to baseline.

[0072] In certain embodiments, the treatment reduces the minimal residual C5 level to about 1.0 microgram / mL or less, for example, about 0.9 microgram / mL or less, 0.8 microgram / mL or less, 0.7 microgram / mL or less, 0.6 microgram / mL or less, 0.5 microgram / mL or less, 0.4 microgram / mL or less, 0.3 microgram / mL or less, 0.2 microgram / mL or less, or 0.1 microgram / mL or less.

[0073] In certain aspects, the treatment reduces classical complement pathway (CCP) activity by at least about 94%, for example, by at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% relative to baseline.

[0074] In certain embodiments, the treatment reduces alternative complement pathway (CAP) activity by at least about 94%, for example, by at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% relative to baseline.

[0075] In certain embodiments, the treatment inhibits mean maximal hemolysis by at least about 75%, such as at least about 76%, at least about 77%, at least about 78%, at least about 79%, at least about 80%, at least about 81%, at least about 82%, at least about 83%, at least about 84%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98% or at least about 99% relative to baseline, as measured by inhibition of ovine erythrocyte hemolysis.

[0076] In certain embodiments, the treatment reduces the level of lactate dehydrogenase (LDH) in the subject to a level lower than about 1.5 times the upper limit of normal (ULN).

[0077] In certain embodiments, subjects previously treated with eculizumab did not have hemolytic attacks. In other embodiments, subjects previously treated with eculizumab had hemolytic attacks.

[0078] In certain embodiments, the treatment reduces the mean maximal C5 mRNA level by at least about 86%, such as at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98% or at least about 99% relative to baseline.

[0079] In one aspect, the treatment reduces the minimum residual C5 level to about 8.0 micrograms / mL or less, such as about 7.5 micrograms / mL or less, about 7.0 micrograms / mL or less, about 6.5 micrograms / mL or less, about 6.0 micrograms / mL or less, about 5.5 micrograms / mL or less, about 5.0 micrograms / mL or less, about 4.5 micrograms / mL or less, about 4.0 micrograms / mL or less, about 3.5 micrograms / mL or less, about 3.0 micrograms / mL or less, about 2.5 micrograms / mL or less, about 2.0 micrograms / mL or less, about 1.5 micrograms / mL or less, about 1.0 micrograms / mL or less, or about 0.5 micrograms / mL or less.

[0080] In certain embodiments, the treatment reduces classical complement pathway (CCP) activity by at least about 98% relative to baseline, such as by at least about 99% relative to baseline.

[0081] In one aspect, the treatment reduces alternative complement pathway (CAP) activity by at least about 98% relative to baseline, such as by at least about 99% relative to baseline.

[0082] In certain embodiments, the treatment inhibits mean maximum hemolysis by at least about 98% relative to baseline, such as by at least about 99% relative to baseline, as measured by inhibition of ovine red blood cell hemolysis.

[0083] In certain embodiments, the level of lactate dehydrogenase (LDH) in the subject is reduced to about 215 - 225 IU / L.

[0084] In one aspect, the dsRNA agent is administered subcutaneously to the subject. In other aspects, eculizumab is administered intravenously to the subject.

[0085] In certain embodiments, the dsRNA agent further comprises a ligand. The ligand can be one or more GalNAc derivatives linked via a divalent or trivalent branched linker. In certain aspects, the ligand is

Chemical formula

[0086] In a further aspect, the ligand is attached to the 3’ end of the sense strand. In certain embodiments, the RNAi agent is conjugated to the ligand as shown in the following schematic

Chemical formula

[0087] In one aspect, the present invention provides a method for treating, e.g., treating for a long period of time, a subject suffering from paroxysmal nocturnal hemoglobinuria (PNH). The method comprises administering, once weekly for 13 weeks, a fixed dose of 200 mg of a double-stranded ribonucleic acid (dsRNA) agent for inhibiting the expression of complement component C5, followed by administering, once weekly, a fixed dose of 400 mg of the dsRNA agent to a subject not receiving eculizumab, thereby treating the subject, wherein the dsRNA agent comprises a sense strand and an antisense strand, wherein the sense strand comprises 5’-asasGfcAfaGfaUfAfUfuUfuuAfuAfaua-3’ (SEQ ID NO: 2876), the antisense strand comprises 5’-usAfsUfuAfuaAfaAfauaUfcUfuGfcuususudTdT-3’ (SEQ ID NO: 2889), wherein a, g, c, and u are 2’-O-methyl (2’-OMe) A, G, C, and U, respectively; Af, Gf, Cf, and Uf are 2’-fluoro A, G, C, and U, respectively; dT is deoxy-thymidine nucleotide; and s is a phosphorothioate bond.

Brief Description of the Drawings

[0088]

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Mode for Carrying Out the Invention

[0089] The present invention provides an iRNA agent that effects cleavage via an RNA-induced silencing complex (RISC) of an RNA transcript of a complement component C5 gene.

[0090] The iRNA of the present invention may comprise an RNA strand (antisense strand) having a region of about 30 nucleotides or less in length, for example, 15 - 30, 15 - 29, 15 - 28, 15 - 27, 15 - 26, 15 - 25, 15 - 24, 15 - 23, 15 - 22, 15 - 21, 15 - 20, 15 - 19, 15 - 18, 15 - 17, 18 - 30, 18 - 29, 18 - 28, 18 - 27, 18 - 26, 18 - 25, 18 - 24, 18 - 23, 18 - 22, 18 - 21, 18 - 20, 19 - 30, 19 - 29, 19 - 28, 19 - 27, 19 - 26, 19 - 25, 19 - 24, 19 - 23, 19 - 22, 19 - 21, 19 - 20, 20 - 30, 20 - 29, 20 - 28, 20 - 27, 20 - 26, 20 - 25, 20 - 24, 20 - 23, 20 - 22, 20 - 21, 21 - 30, 21 - 29, 21 - 28, 21 - 27, 21 - 26, 21 - 25, 21 - 24, 21 - 23, or 21 - 22 nucleotides in length, and this region is substantially complementary to at least a part of the mRNA transcript of the C5 gene.

[0091] In certain embodiments, the iRNA of the invention comprises an RNA strand (antisense strand) having a region of at least 19 contiguous nucleotides that is substantially complementary to at least a portion of the mRNA transcript of the complement component C5 gene, and a longer length, for example, up to 66 nucleotides, for example, lengths including 36 - 66, 26 - 36, 25 - 36, 31 - 60, 22 - 43, 27 - 53 nucleotides. These iRNAs having longer antisense strands preferably comprise a second RNA strand (sense strand) that is 20 - 60 nucleotides in length, where the sense and antisense strands form a double-stranded region of 18 - 30 contiguous nucleotides.

[0092] The use of these iRNAs enables targeted degradation of the mRNA of the mammalian C5 gene. In particular, very low doses of C5 iRNA can specifically and efficiently mediate RNA interference (RNAi) and result in a significant inhibition of C5 gene expression. The inventors have demonstrated that iRNAs targeting C5 can mediate RNAi in vitro and in vivo and result in a significant inhibition of C5 gene expression. Thus, methods and compositions comprising these iRNAs are useful for treating subjects who can benefit from a decrease in the level and / or activity of C5 protein, such as subjects suffering from a disease associated with complement component C5, such as paroxysmal nocturnal hemoglobinuria (PNH).

[0093] The present invention also provides methods and combination therapies for treating subjects suffering from a disorder, such as paroxysmal nocturnal hemoglobinuria (PNH), atypical hemolytic uremic syndrome (aHUS), neuromyelitis optica (NMO), and myasthenia gravis, which can benefit from inhibiting or reducing the expression of the C5 gene using an iRNA composition that effects cleavage via an RNA-induced silencing complex (RISC) of an RNA transcript of the complement component C5 gene.

[0094] The present invention also provides a method for preventing at least one symptom, such as hemolysis, in a subject suffering from a disorder, such as paroxysmal nocturnal hemoglobinuria (PNH), atypical hemolytic uremic syndrome (aHUS), neuromyelitis optica (NMO), and myasthenia gravis, which can benefit from inhibiting or reducing the expression of the C5 gene. The present invention further provides an iRNA composition that results in cleavage of an RNA transcript of the complement component C5 gene via an RNA-induced silencing complex (RISC). The C5 gene can be present within a cell, such as a cell within a subject, such as a human.

[0095] The combination therapy of the present invention comprises administering to a subject suffering from a disease associated with complement component C5, the RNAi agent of the present invention and an anti-complement component C5 antibody, or an antigen-binding fragment thereof, such as eculizumab, as a further therapeutic agent. The combination therapy of the present invention uses the iRNA agent of the present invention to target C5 mRNA to reduce (e.g., by about 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or about 99%) the C5 level in the subject, and thus reduce the therapeutically (or prophylactically) effective amount of eculizumab required to treat the subject, thereby reducing the treatment cost and enabling an easier and more convenient method of administering eculizumab, such as subcutaneous administration.

[0096] The following detailed description discloses methods for making and using compositions containing iRNA for inhibiting the expression of the C5 gene, as well as compositions, uses, and methods for treating subjects suffering from diseases and disorders that can benefit from inhibition and / or reduction of the expression of this gene.

[0097] I. Definitions To make the present invention more readily understood, several terms are first defined. Further, it should be noted that whenever a value or range of values of a variable is recited, intermediate values and ranges within the recited values are also intended to be part of the present invention.

[0098] The articles "a" and "an" are used herein to refer to one or more (i.e., at least one) of the grammatical objects of the article. By way of example, "an element" means one element or more than one element, e.g., a plurality of elements.

[0099] The term "including" is used herein to mean the phrase "including but not limited to" and is used synonymously with this phrase.

[0100] The term "or" is used herein to mean the term "and / or" unless the context clearly indicates otherwise and is used synonymously with this term.

[0101] As used herein, "complement component C5", which is used synonymously with the term "C5", refers in the art to well-known genes and polypeptides also known as CPAMD4, C3 and PZP-like alpha-2-macroglobulin domain-containing protein, anaphylatoxin C5a analog, hemolytic complement (Hc), and complement C5. The sequence of the human C5 mRNA transcript can be found, for example, in GenBank accession number GI:38016946 (NM_001735.2; SEQ ID NO:1). The sequence of the rhesus monkey C5 mRNA can be found, for example, in GenBank accession number GI:297270262 (XM_001095750.2; SEQ ID NO:2). The sequence of the mouse C5 mRNA can be found, for example, in GenBank accession number GI:291575171 (NM_010406.2; SEQ ID NO:3). The sequence of the rat C5 mRNA can be found, for example, in GenBank accession number GI:392346248 (XM_345342.4; SEQ ID NO:4). Further examples of C5 mRNA sequences are readily available using publicly available databases such as GenBank.

[0102] As used herein, the term "C5" also refers to variations in the native DNA sequence of the C5 gene, such as single nucleotide polymorphisms in the C5 gene. Many SNPs in the C5 gene have been identified and can be found, for example, in NCBI dbSNP (see, e.g., ncbi.nlm.nih.gov / snp). Non-limiting examples of SNPs within the C5 gene can be found in NCBI dbSNP accession numbers rs121909588 and rs121909587.

[0103] As used herein, "target sequence" refers to a contiguous portion of the nucleotide sequence of an mRNA molecule formed upon transcription of the C5 gene, including the mRNA that is the product of RNA processing of the primary transcript. In one embodiment, the target portion of the sequence will be at least long enough to function as a substrate for iRNA-directed cleavage at or near the corresponding portion of the nucleotide sequence of the mRNA molecule formed upon transcription of the C5 gene.

[0104] The target sequence can be about 9 to 36 nucleotides in length, such as about 15 to 30 nucleotides in length. For example, the target sequence can be about 15 to 30 nucleotides, 15 - 29, 15 - 28, 15 - 27, 15 - 26, 15 - 25, 15 - 24, 15 - 23, 15 - 22, 15 - 21, 15 - 20, 15 - 19, 15 - 18, 15 - 17, 18 - 30, 18 - 29, 18 - 28, 18 - 27, 18 - 26, 18 - 25, 18 - 24, 18 - 23, 18 - 22, 18 - 21, 18 - 20, 19 - 30, 19 - 29, 19 - 28, 19 - 27, 19 - 26, 19 - 25, 19 - 24, 19 - 23, 19 - 22, 19 - 21, 19 - 20, 20 - 30, 20 - 29, 20 - 28, 20 - 27, 20 - 26, 20 - 25, 20 - 24, 20 - 23, 20 - 22, 20 - 21, 21 - 30, 21 - 29, 21 - 28, 21 - 27, 21 - 26, 21 - 25, 21 - 24, 21 - 23, or 21 - 22 nucleotides in length. Ranges and lengths intermediate to those described above are also contemplated as part of the present invention.

[0105] As used herein, the term "strand containing an array" refers to an oligonucleotide containing a nucleotide strand represented by an array shown using standard nucleotide nomenclature.

[0106] "G", "C", "A", and "U" each generally represent nucleotides containing the bases guanine, cytosine, adenine, and uracil, respectively. However, it will be understood that the terms "ribonucleotide" or "nucleotide" can also refer to modified nucleotides, or surrogate replacement moieties, as further detailed below (see, e.g., Table 2). One of ordinary skill in the art will fully recognize that guanine, cytosine, adenine, and uracil may be replaced by other moieties without significantly altering the base pairing properties of the oligonucleotide containing nucleotides having such replacement moieties. For example, without limitation, a nucleotide containing inosine as its base can base pair with a nucleotide containing adenine, cytosine, or uracil. Thus, a nucleotide containing uracil, guanine, or adenine may be replaced, for example, by a nucleotide containing inosine in the nucleotide sequence of the dsRNA characterized in the present invention. In another example, some adenines and cytosines in an oligonucleotide may be replaced by guanine and uracil, respectively, to form wobble G-U base pairing with a target mRNA. Sequences containing such replacement moieties are suitable for the compositions and methods encompassed by the present invention.

[0107] The terms "iRNA", "RNAi agent", "iRNA agent", and "RNA interference agent", which are used interchangeably herein, refer to an agent that contains RNA and mediates the targeted cleavage of an RNA transcript via the RNA-induced silencing complex (RISC) pathway, as defined herein. iRNA leads to sequence-specific degradation of mRNA by a process known as RNA interference (RNAi). iRNA modulates (e.g., inhibits) the expression of C5 in cells, e.g., in cells of a mammalian subject.

[0108] In one embodiment, the RNAi agent of the present invention includes a single-stranded RNA that interacts with a target RNA sequence, for example, a C5 target mRNA sequence, to direct cleavage of the target RNA. Without wishing to be bound by theory, it is believed that long double-stranded RNA introduced into cells is degraded into siRNA by type III endonucleases known as Dicer (Sharp et al. (2001) Genes Dev. 15:485). Dicer, a ribonuclease III-like enzyme, processes dsRNA into short interfering RNAs of 19-23 base pairs with characteristic two-base 3' overhangs (Bernstein, et al., (2001) Nature 409:363). Next, the siRNA is incorporated into the RNA-induced silencing complex (RISC), where one or more helicases unwind the siRNA duplex, allowing the complementary antisense strand to direct target recognition (Nykanen, et al., (2001) Cell 107:309). Upon binding to the appropriate target mRNA, one or more endonucleases in the RISC cleave the target to induce silencing (Elbashir, et al., (2001) Genes Dev. 15:188). Here, in one aspect, the present invention relates to a single-stranded RNA (siRNA) that is generated in cells and promotes the formation of the RISC complex that results in silencing of the target gene, namely, the C5 gene. Accordingly, the term "siRNA" is also used herein to refer to the RNAi described above.

[0109] In another embodiment, the RNAi agent can be a single-stranded siRNA that is introduced into a cell or organism to inhibit a target mRNA. The single-stranded RNAi agent binds to the RISC endonuclease Argonaute 2, which then cleaves the target mRNA. Single-stranded siRNAs are generally 15 to 30 nucleotides in length and are chemically modified. The design and testing of single-stranded siRNAs are described in U.S. Patent No. 8,101,348 and Lima et al., (2012) Cell 150:883-894, the entire contents of each of which are incorporated herein by reference. Any of the antisense nucleotide sequences described herein can be used as a single-stranded siRNA that is chemically modified by the methods described herein or as described in Lima et al., (2012) Cell 150;:883-894.

[0110] In another embodiment, the "iRNA" for use in the compositions, uses, and methods of the present invention is double-stranded RNA, referred to herein as "double-stranded RNAi agent", "double-stranded RNA (dsRNA) molecule", "dsRNA agent", or "dsRNA". The term "dsRNA" refers to a complex of ribonucleic acid molecules having a double-stranded structure comprising two anti-parallel and substantially complementary nucleic acid strands that are shown to have "sense" and "antisense" orientations with respect to a target RNA, i.e., the C5 gene. In certain embodiments of the invention, the double-stranded RNA (dsRNA) causes degradation of a target RNA, e.g., mRNA, by a post-transcriptional gene silencing mechanism referred to herein as RNA interference or RNAi.

[0111] Generally, most of the nucleotides of each strand of a dsRNA molecule are ribonucleotides, but as described in detail herein, each or both strands may also include one or more non-ribonucleotides, such as deoxyribonucleotides and / or modified nucleotides. Further, as used herein, an "RNAi agent" may include ribonucleotides having chemical modifications; an RNAi agent may include substantial modifications in multiple nucleotides. As used herein, the term "modified nucleotide" independently refers to a nucleotide having a modified sugar moiety, a modified internucleotide linkage, and / or a modified nucleobase. Thus, the term modified nucleotide encompasses substitutions, additions, or removals of, for example, functional groups or atoms to the internucleoside linkage, the sugar moiety, or the nucleobase. Suitable modifications for use in the agents of the present invention include any type of modification disclosed herein or known in the art. Any such modification when used in an siRNA-type molecule is encompassed by an "RNAi agent" for the purposes of this specification and the claims.The double-stranded region may be of any length that enables specific degradation of a desired target RNA via the RISC pathway, and may be in the range of about 9 to 36 base pairs in length, for example, in the range of about 15 to 30 base pairs in length, for example, about 15 to 30, 15 to 29, 15 to 28, 15 to 27, 15 to 26, 15 to 25, 15 to 24, 15 to 23, 15 to 22, 15 to 21, 15 to 20, 15 to 19, 15 to 18, 15 to 17, 18 to 30, 18 to 29, 18 to 28, 18 to 27, 18 to 26, 18 to 25, 18 to 24, 18 to 23, 18 to 22, 18 to 21, 18 to 20, 19 to 30, 19 to 29, 19 to 28, 19 to 27, 19 to 26, 19 to 25, 19 to 24, 19 to 23, 19 to 22, 19 to 21, 19 to 20, 20 to 30, 20 to 29, 20 to 28, 20 to 27, 20 to 26, 20 to 25, 20 to 24, 20 to 23, 20 to 22, 20 to 21, 21 to 30, 21 to 29, 21 to 28, 21 to 27, 21 to 26, 21 to 25, 21 to 24, 21 to 23, or 21 to 22 base pairs in length, and may be, for example, about 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, or 36 base pairs in length. Ranges and lengths intermediate to those described above are also considered to be part of the present invention.

[0112] The two strands forming the double-stranded structure may be different portions of one larger RNA molecule, or they may be separate RNA molecules. When the two strands are part of one larger molecule and are thus joined by a continuous strand of nucleotides between the 3' end of one strand forming the double-stranded structure and the 5' end of the other strand, the joined RNA strands are referred to as a "hairpin loop". The hairpin loop may contain at least one unpaired nucleotide. In certain embodiments, the hairpin loop may contain at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 20, at least 23, or more unpaired nucleotides.

[0113] When two substantially complementary strands of dsRNA are composed of separate RNA molecules, those molecules may or may not be covalently linked. When the two strands are covalently linked by means other than a continuous strand of nucleotides between the 3' end of one strand forming a double-stranded structure and the 5' end of the other strand, the linkage structure is referred to as a "linker". The RNA strands can have the same or different numbers of nucleotides. The maximum number of base pairs is the number of nucleotides in the shortest strand of the dsRNA minus the number of overhangs present in the duplex. In addition to the double-stranded structure, the RNAi agent can include one or more nucleotide overhangs.

[0114] In one embodiment, the RNAi agent of the invention is a 24 - 30 nucleotide dsRNA that interacts with a target RNA sequence, e.g., a C5 target mRNA sequence, to direct cleavage of the target RNA. Without wishing to be bound by theory, long double-stranded RNAs introduced into cells are degraded into siRNAs by a type III endonuclease known as Dicer (Sharp et al. (2001) Genes Dev. 15:485). Dicer, a ribonuclease III-like enzyme, processes dsRNA into short interfering RNAs of 19 - 23 base pairs with characteristic two-base 3' overhangs (Bernstein, et al., (2001) Nature 409:363). The siRNAs are then incorporated into the RNA-induced silencing complex (RISC), where one or more helicases unwind the siRNA duplex, allowing the complementary antisense strand to enable target recognition (Nykanen, et al., (2001) Cell 107:309). Upon binding to the appropriate target mRNA, one or more endonucleases in the RISC cleave the target to induce silencing (Elbashir, et al., (2001) Genes Dev. 15:188).

[0115] As used herein, the term "nucleotide overhang" refers to at least one unpaired nucleotide that protrudes from the double-stranded structure of an iRNA, such as dsRNA. For example, when the 3' end of one strand of dsRNA extends beyond the 5' end of the other strand or vice versa, a nucleotide overhang exists. The dsRNA can include an overhang of at least one nucleotide; alternatively, the overhang can include at least two nucleotides, at least three nucleotides, at least four nucleotides, at least five nucleotides or more. The nucleotide overhang can include or consist of nucleotide / nucleoside analogs including deoxynucleotides / nucleosides. The overhang can be the sense strand, the antisense strand or any combination thereof. Further, the nucleotides of the overhang can be present at the 5' end, 3' end or both ends of either the antisense strand or the sense strand of the dsRNA.

[0116] In one embodiment, the antisense strand of the dsRNA has an overhang of 1 to 10 nucleotides, such as 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 nucleotides, at the 3' end and / or 5' end. In one embodiment, the sense strand of the dsRNA has an overhang of 1 to 10 nucleotides, such as 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 nucleotides, at the 3' end and / or 5' end. In another embodiment, one or more of the nucleotides in the overhang are replaced with nucleoside phosphorothioates.

[0117] In certain embodiments, the overhangs in the sense strand or the antisense strand, or both, can include an extended length greater than 10 nucleotides in length, such as 1 to 30 nucleotides, 2 to 30 nucleotides, 10 to 30 nucleotides, or 10 to 15 nucleotides in length. In certain embodiments, the extended overhang is on the double-stranded sense strand. In certain embodiments, the extended overhang is present at the 3' end of the double-stranded sense strand. In certain embodiments, the extended overhang is present at the 5' end of the double-stranded sense strand. In certain embodiments, the extended overhang is on the double-stranded antisense strand. In certain embodiments, the extended overhang is present at the 3' end of the double-stranded antisense strand. In certain embodiments, the extended overhang is present at the 5' end of the double-stranded antisense strand. In certain embodiments, one or more of the nucleotides in the overhang are replaced with nucleoside phosphorothioates. In certain embodiments, the overhang includes self-complementary portions such that it is capable of forming a hairpin structure that is stable under physiological conditions.

[0118] "Blunt" or "blunt end" means that there are no unpaired nucleotides at the corresponding end of the double-stranded RNAi agent, i.e., there is no nucleotide overhang. A "blunt end" RNAi agent is double-stranded over its entire length, i.e., it is a dsRNA having no nucleotide overhangs at either end of the molecule. The RNAi agents of the present invention include RNAi agents having a nucleotide overhang at one end (i.e., an agent having one overhang and one blunt end) or RNAi agents having nucleotide overhangs at both ends.

[0119] The term "antisense strand" or "guide strand" refers to a strand of an iRNA, e.g., a dsRNA, that contains a region substantially complementary to a target sequence, e.g., the C5 mRNA. As used herein, the term "region of complementarity" refers to a region of the antisense strand that is substantially complementary to a sequence, e.g., a target sequence, e.g., the C5 nucleotide sequence, as defined herein. If the region of complementarity is not completely complementary to the target sequence, mismatches may be present in the internal or terminal regions of the molecule. Generally, most tolerated mismatches are present in the terminal regions, e.g., 5, 4, 3, or 2 nucleotides at the 5' end and / or 3' end of the iRNA.

[0120] As used herein, the term "sense strand" or "passenger strand" refers to a strand of an iRNA that contains a region substantially complementary to the region of the antisense strand, as the term is defined herein.

[0121] As used herein, the term "cleavage region" refers to the region located immediately adjacent to the cleavage site. The cleavage site is the site at which cleavage occurs in the target. In certain embodiments, the cleavage region contains 3 bases immediately adjacent to the cleavage site at either end of the cleavage site. In certain embodiments, the cleavage region contains 2 bases immediately adjacent to the cleavage site at either end of the cleavage site. In certain embodiments, the cleavage site occurs specifically at the site joined by nucleotides 10 and 11 of the antisense strand, and the cleavage region contains nucleotides 11, 12, and 13.

[0122] As used herein, unless otherwise indicated, the term "complementary," when used to describe a first nucleotide sequence in relation to a second nucleotide sequence, as understood by one of ordinary skill in the art, refers to the ability of an oligonucleotide or polynucleotide containing the first nucleotide sequence to hybridize with an oligonucleotide or polynucleotide containing the second nucleotide sequence and form a double-stranded structure under certain conditions. Such conditions can be, for example, stringent conditions, where stringent conditions can include 400 mM NaCl, 40 mM PIPES (pH 6.4), 1 mM EDTA, 12-16 hours at 50°C or 70°C, followed by washing (see, e.g., "Molecular Cloning: A Laboratory Manual, Sambrook, et al. (1989) Cold Spring Harbor Laboratory Press"). Other conditions can be applied, such as physiologically relevant conditions that can occur within an organism. One of ordinary skill in the art will be able to determine the most appropriate set of conditions for testing the complementarity of the two sequences according to the ultimate use of the hybridized nucleotides.

[0123] Among the iRNAs described herein, for example, the complementary sequences in dsRNA include base pairing over the entire length of one or both nucleotide sequences of an oligonucleotide or polynucleotide containing a first nucleotide sequence to an oligonucleotide or polynucleotide containing a second nucleotide sequence. Such sequences may be referred to herein as "perfectly complementary" to each other. However, when a first sequence is referred to herein as "substantially complementary" to a second sequence, the two sequences may be perfectly complementary, or, while retaining the ability to hybridize under conditions optimal for their ultimate application, e.g., inhibition of gene expression via the RISC pathway, they may form one or more, but generally 5 or fewer, 4 or fewer, 3 or fewer, or 2 or fewer mismatched base pairs when hybridized to a duplex of up to 30 base pairs. However, if two oligonucleotides are designed to form one or more single-stranded overhangs after hybridization, such overhangs shall not be considered mismatches with respect to the determination of complementarity. For example, for the purposes described herein, a dsRNA containing one 21-nucleotide-long oligonucleotide and another 23-nucleotide-long oligonucleotide may be referred to as "perfectly complementary" if the longer oligonucleotide contains a 21-nucleotide sequence that is perfectly complementary to the shorter oligonucleotide.

[0124] As used herein, "complementary" sequences can also include, or can be entirely formed from, base pairs formed from non-Watson-Crick base pairs and / or non-natural and modified nucleotides, so long as the above requirements regarding their ability to hybridize are met. Such non-Watson-Crick base pairs include, but are not limited to, G:U wobbles or Hoogsteen-type base pairs.

[0125] As used herein, the terms "complementary", "fully complementary" and "substantially complementary" can be used in connection with matching bases between the sense and antisense strands of a dsRNA, or between the antisense strand of a dsRNA and a target sequence, as understood from the context of their use.

[0126] As used herein, a polynucleotide "substantially complementary to at least a portion of" a messenger RNA (mRNA) refers to a polynucleotide that is substantially complementary to a continuous portion of the target mRNA (e.g., an mRNA encoding PCSK9) that includes the 5'UTR, open reading frame (ORF), or 3'UTR. For example, a polynucleotide is complementary to at least a portion of a PCSK9 mRNA if its sequence is substantially complementary to a continuous portion of the mRNA encoding PCSK9.

[0127] Thus, in certain embodiments, the sense strand polynucleotides and antisense polynucleotides disclosed herein are fully complementary to a complement component C5 gene sequence.

[0128] In one embodiment, the antisense polynucleotides disclosed herein are fully complementary to a target complement component C5 sequence. In other embodiments, the antisense polynucleotides disclosed herein are substantially complementary to a target complement component C5 sequence and are at least about 80% complementary over their entire length to an equivalent region of the nucleotide sequence of SEQ ID NO: 1, or a fragment of SEQ ID NO: 1, e.g., about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% complementary, and include a continuous nucleotide sequence.

[0129] In other embodiments, the antisense polynucleotides disclosed herein are substantially complementary to the target complement component C5 sequence and are at least about 80% complementary over their entire length to any one of the sense strand nucleotide sequences in any one of Tables 3, 4, 5, 6, 18, 19, 20, 21, and 23, or to any one of the fragments of any one of the antisense strand nucleotide sequences in any one of Tables 3, 4, 5, 6, 18, 19, 20, 21, and 23, for example, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% complementary, and include a continuous nucleotide sequence.

[0130] In one embodiment, the RNAi agent of the present invention is complementary to the target complement component C5 sequence and includes a sense strand that is substantially complementary to an antisense polynucleotide that is at least about 80% complementary over its entire length to any one of the antisense strand nucleotide sequences in any one of Tables 3, 4, 5, 6, 18, 19, 20, 21, and 23, or to any one of the fragments of any one of the antisense strand nucleotide sequences in any one of Tables 3, 4, 5, 6, 18, 19, 20, 21, and 23, for example, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% complementary, and includes a continuous nucleotide sequence.

[0131] Generally, most of the nucleotides of each strand are ribonucleotides, but as described in detail herein, each or both strands may also include one or more non-ribonucleotides, such as deoxyribonucleotides and / or modified nucleotides. Further, "iRNA" may include ribonucleotides having chemical modifications. Such modifications may include any type of modification disclosed herein or known in the art. Any such modification when used in an iRNA molecule is encompassed by "iRNA" for the purposes of this specification and the claims.

[0132] In one aspect of the present invention, the agent for use in the methods and compositions of the present invention is a single-stranded antisense RNA molecule that inhibits target mRNA via an antisense inhibition mechanism. The single-stranded antisense RNA molecule is complementary to the sequence in the target mRNA. The single-stranded antisense oligonucleotide can inhibit translation stoichiometrically by base pairing with the mRNA and physically interfering with the translation machinery (see Dias, N. et al., (2002) Mol Cancer Ther 1:347-355). The single-stranded antisense RNA molecule is about 15 to about 30 nucleotides in length and can have a sequence complementary to the target sequence. For example, the single-stranded antisense RNA molecule can include a sequence that is at least about 15, 16, 17, 18, 19, 20, or more consecutive nucleotides from any one of the antisense sequences described herein.

[0133] The term "lipid nanoparticle" or "LNP" refers to a vesicle that includes a lipid layer encapsulating a nucleic acid molecule, such as a pharmaceutically active molecule such as an iRNA or a plasmid from which the iRNA is transcribed. LNPs are described, for example, in U.S. Patent Nos. 6,858,225, 6,815,432, 8,158,601, and 8,058,069, the entire contents of each of which are incorporated herein by reference.

[0134] As used herein, "subject" is an animal such as a mammal including primates (humans, non-human primates such as monkeys and chimpanzees, etc.), non-primates (cows, pigs, camels, llamas, horses, goats, rabbits, sheep, hamsters, guinea pigs, cats, dogs, rats, mice, horses, and whales, etc.), or birds (such as ducks or geese). In one embodiment, the subject is a human being who is being treated or evaluated for a disease, disorder or condition that may benefit from a decrease in the expression of C5 as described herein; a human being at risk of a disease, disorder or condition that may benefit from a decrease in the expression of C5; a human being suffering from a disease, disorder or condition that may benefit from a decrease in the expression of C5; and / or a human being such as a human being who is being treated for a disease, disorder or condition that may benefit from a decrease in the expression of C5.

[0135] As used herein, the term "treating" or "treatment" refers to, but is not limited to, the alleviation or improvement of one or more symptoms (such as hemolysis and / or chronic inflammation) associated with unwanted complement pathway activation; a decrease in the degree of unwanted complement pathway activation; stabilization of the state of chronic inflammation and / or hemolysis (i.e., not getting worse); improvement or alleviation of unwanted complement pathway activation (such as chronic inflammation and / or hemolysis) whether detectable or undetectable. "Treatment" can also mean an extension of the survival period compared to the predicted survival period without treatment.

[0136] The term "lowering" in the context of complement component C5 or a disease marker or symptom in a subject refers to a statistically significant decrease in such levels. The decrease can be, for example, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or more, and is preferably lowered to a level recognized as within the normal range of an individual without such a disorder.

[0137] As used herein, "prevent" or "preventing" when used in connection with a disease, disorder or condition in which a benefit can be obtained from a decrease in the expression of the C5 gene, means that the subject has a reduced likelihood of developing symptoms associated with such disease, disorder or condition, or a decrease in the frequency and / or duration of symptoms associated with such disease, disorder or condition, such as symptoms of unwanted complement activation such as chronic inflammation, hemolysis and / or thrombosis. The likelihood of developing thrombosis is reduced, for example, if an individual having one or more risk factors for thrombosis does not develop thrombosis, or develops a less severe thrombosis compared to a group having the same risk factors and not receiving the treatment described herein. Not developing a disease, disorder or condition, or a decrease in the occurrence of symptoms associated with such disease, disorder or condition (e.g., at least about 10% on a clinically recognized scale of such disease or disorder), or a delayed appearance of symptoms (e.g., by only a few days, weeks, months or years) is considered effective prevention.

[0138] As used herein, the term "disease associated with complement component C5" refers to a disease or disorder caused by or associated with complement activation. Such diseases are typically associated with inflammation and / or activation of the immune system, such as lysis by the membrane attack complex, anaphylaxis, and / or hemolysis. Non-limiting examples of diseases associated with complement component C5 include paroxysmal nocturnal hemoglobinuria (PNH), atypical hemolytic uremic syndrome (aHUS), asthma, rheumatoid arthritis (RA); antiphospholipid antibody syndrome; lupus nephritis; ischemia-reperfusion injury; typical or infectious hemolytic uremic syndrome (tHUS); dense deposit glomerulonephritis (DDD); neuromyelitis optica (NMO); multifocal motor neuropathy (MMN); multiple sclerosis (MS); macular degeneration (e.g., age-related macular degeneration (AMD)); hemolysis, elevated liver escape enzymes, and low platelets (HELLP) syndrome; thrombotic thrombocytopenic purpura (TTP); spontaneous abortion; microscopic immunologic vasculitis; epidermolysis bullosa; habitual abortion; pregnancy-induced hypertension nephropathy, traumatic brain injury, myasthenia gravis, cold agglutinin disease, dermatomyositis, bullous pemphigoid, Shiga toxin-producing Escherichia coli (E.hemolytic uremic syndrome associated with Escherichia coli (E. coli), C3 glomerulopathy, antineutrophil cytoplasmic antibody-associated vasculitis (e.g., granulomatosis with polyangiitis (previously known as Wegener's granulomatosis), Churg-Strauss syndrome, and microscopic polyangiitis), humoral and vascular transplant rejection, graft dysfunction, myocardial infarction (e.g., tissue injury and ischemia in myocardial infarction), allotransplantation, sepsis (e.g., poor outcome in sepsis), coronary artery disease, dermatomyositis, Graves' disease, atherosclerosis, Alzheimer's disease, systemic inflammatory response sepsis, septic shock, spinal cord injury, glomerulonephritis, Hashimoto's thyroiditis, type 1 diabetes, psoriasis, pemphigus, autoimmune hemolytic anemia (AIHA), ITP, Goodpasture's syndrome, Doge disease, antiphospholipid syndrome (APS), catastrophic APS (CAPS), cardiovascular disease, myocarditis, cerebrovascular disorder, peripheral (e.g., skeletal) vascular disorder, renal vascular disorder, mesenteric / intestinal vascular disorder, vasculitis, Henoch-Schönlein purpura nephritis, systemic lupus erythematosus-related vasculitis, vasculitis associated with rheumatoid arthritis, immune complex vasculitis, Takayasu disease, dilated cardiomyopathy, diabetic angiopathy, Kawasaki disease (arteritis), venous gas embolism (VGE), and restenosis after stent placement, rotational atherectomy, membranous nephropathy, Guillain-Barré syndrome, and percutaneous transluminal coronary angioplasty (PTCA) (see, e.g., Holers (2008) Immunological Reviews 223:300-316; Holers and Thurman (2004) Molecular Immunology 41:147-152; U.S. Patent Application Publication No. 20070172483).

[0139] In one embodiment, the disease associated with complement component C5 is paroxysmal nocturnal hemoglobinuria (PNH). PNH can be classical PNH or another myelodysplastic syndrome and / or myelodysplasia (MDS), e.g., PNH in the case of cytopenia. In another embodiment, the disease associated with complement component C5 is atypical hemolytic uremic syndrome (aHUS). In another embodiment, the disease associated with complement component C5 is neuromyelitis optica (NMO). In yet another embodiment, the disease associated with complement component C5 is myasthenia gravis.

[0140] II. iRNA of the Present Invention The present invention provides an iRNA that inhibits the expression of the complement component C5 gene. In one embodiment, the iRNA agent is a double-stranded ribonucleic acid (dsRNA) molecule for inhibiting the expression of the C5 gene in cells, such as cells in a subject suffering from a disease associated with complement component C5, e.g., PNH, such as a mammal such as a human. The dsRNA includes an antisense strand having a complementary region that is at least partially complementary to at least a portion of the mRNA formed during the expression of the C5 gene. The complementary region is about 30 nucleotides or less in length (e.g., about 30, 29, 28, 27, 26, 25, 24, 23, 22, 21, 20, 19, or 18 nucleotides or less in length). When contacted with cells expressing the C5 gene, the iRNA inhibits the expression of the C5 gene (e.g., the C5 gene of a human, primate, non-primate, or bird) by at least about 10% when assayed by, for example, a protein-based method such as PCR or branched DNA (bDNA) assay, or immunofluorescence analysis using, for example, Western blot or flow cytometry techniques.

[0141] The dsRNA includes two RNA strands that are complementary and hybridize to form a double-stranded structure under the conditions in which the dsRNA is used. One strand (antisense strand) of the dsRNA includes a complementary region that is substantially complementary, generally fully complementary, to the target sequence. The target sequence may be derived from the sequence of the mRNA formed during the expression of the C5 gene. The other strand (sense strand) includes a region complementary to the antisense strand and is adapted to hybridize with the two strands to form a double-stranded structure when combined under suitable conditions. As described elsewhere herein and as known in the art, the complementary sequences of the dsRNA may also be included as self-complementary regions of a single nucleic acid molecule rather than on separate oligonucleotides.

[0142] Generally, the double-stranded structure has a length of about 15 to 30 base pairs, for example, about 15 to 29, 15 to 28, 15 to 27, 15 to 26, 15 to 25, 15 to 24, 15 to 23, 15 to 22, 15 to 21, 15 to 20, 15 to 19, 15 to 18, 15 to 17, 18 to 30, 18 to 29, 18 to 28, 18 to 27, 18 to 26, 18 to 25, 18 to 24, 18 to 23, 18 to 22, 18 to 21, 18 to 20, 19 to 30, 19 to 29, 19 to 28, 19 to 27, 19 to 26, 19 to 25, 19 to 24, 19 to 23, 19 to 22, 19 to 21, 19 to 20, 20 to 30, 20 to 29, 20 to 28, 20 to 27, 20 to 26, 20 to 25, 20 to 24, 20 to 23, 20 to 22, 20 to 21, 21 to 30, 21 to 29, 21 to 28, 21 to 27, 21 to 26, 21 to 25, 21 to 24, 21 to 23, or 21 to 22 base pairs in length. The intermediate ranges and lengths within the ranges and lengths described above are also considered to be part of the present invention.

[0143] Similarly, the complementary region of the target sequence has a length of about 15 to 30 nucleotides, for example, about 15 to 29, 15 to 28, 15 to 27, 15 to 26, 15 to 25, 15 to 24, 15 to 23, 15 to 22, 15 to 21, 15 to 20, 15 to 19, 15 to 18, 15 to 17, 18 to 30, 18 to 29, 18 to 28, 18 to 27, 18 to 26, 18 to 25, 18 to 24, 18 to 23, 18 to 22, 18 to 21, 18 to 20, 19 to 30, 19 to 29, 19 to 28, 19 to 27, 19 to 26, 19 to 25, 19 to 24, 19 to 23, 19 to 22, 19 to 21, 19 to 20, 20 to 30, 20 to 29, 20 to 28, 20 to 27, 20 to 26, 20 to 25, 20 to 24, 20 to 23, 20 to 22, 20 to 21, 21 to 30, 21 to 29, 21 to 28, 21 to 27, 21 to 26, 21 to 25, 21 to 24, 21 to 23, or 21 to 22 nucleotides in length. The intermediate ranges and lengths within the ranges and lengths described above are also considered to be part of the present invention.

[0144] In certain embodiments, the dsRNA is from about 15 to about 20 nucleotides in length, or from about 25 to about 30 nucleotides in length. Generally, the dsRNA is long enough to function as a substrate for the Dicer enzyme. For example, it is well known in the art that dsRNA longer than about 21-23 nucleotides in length can function as a substrate for Dicer. As will also be appreciated by those skilled in the art, the region of the RNA targeted for cleavage is, in most cases, part of a larger RNA molecule, often an mRNA molecule. When relevant, a "portion" of the mRNA target is a continuous sequence of the mRNA target that is long enough to be capable of being a substrate for RNAi-directed cleavage (i.e., cleavage via the RISC pathway).

[0145] The double-stranded region is the primary functional part of the dsRNA, for example, about 9 to 36 base pairs, such as about 10 to 36, 11 to 36, 12 to 36, 13 to 36, 14 to 36, 15 to 36, 9 to 35, 10 to 35, 11 to 35, 12 to 35, 13 to 35, 14 to 35, 15 to 35, 9 to 34, 10 to 34, 11 to 34, 12 to 34, 13 to 34, 14 to 34, 15 to 34, 9 to 33, 10 to 33, 11 to 33, 12 to 33, 13 to 33, 14 to 33, 15 to 33, 9 to 32, 10 to 32, 11 to 32, 12 to 32, 13 to 32, 14 to 32, 15 to 32, 9 to 31, 10 to 31, 11 to 31, 12 to 31, 13 to 32, 14 to 31, 15 to 31, 15 to 30, 15 to 29, 15 to 28, 15 to 27, 15 to 26, 15 to 25, 15 to 24, 15 to 23, 15 to 22, 15 to 21, 15 to 20, 15 to 19, 15 to 18, 15 to 17, 18 to 30, 18 to 29, 18 to 28, 18 to 27, 18 to 26, 18 to 25, 18 to 24, 18 to 23, 18 to 22, 18 to 21, 18 to 20, 19 to 30, 19 to 29, 19 to 28, 19 to 27, 19 to 26, 19 to 25, 19 to 24, 19 to 23, 19 to 22, 19 to 21, 19 to 20, 20 to 30, 20 to 29, 20 to 28, 20 to 27, 20 to 26, 20 to 25, 20 to 24, 20 to 23, 20 to 22, 20 to 21, 21 to 30, 21 to 29, 21 to 28, 21 to 27, 21 to 26, 21 to 25, 21 to 24, 21 to 23, or a double-stranded region of 21 to 22 base pairs. Those skilled in the art will also recognize this. Here, in one embodiment, a complex of an RNA molecule or an RNA molecule having a double-stranded region of more than 30 base pairs is processed to a functional double-stranded of, for example, 15 to 30 base pairs to target the desired RNA for cleavage, and such a complex is dsRNA. Thus, those skilled in the art will recognize that in one embodiment, miRNA is dsRNA. In another embodiment, the dsRNA is not a natural miRNA. In another embodiment, the iRNA agent useful for targeting the expression of C5 is not generated in the target cells by cleavage of larger dsRNAs.

[0146] The dsRNA described in this specification may further include one or more single-stranded nucleotide overhangs, for example, 1, 2, 3, or 4 nucleotides. dsRNA having at least one nucleotide overhang may have unexpectedly excellent inhibitory properties compared to its blunt-end equivalent. The nucleotide overhang may comprise or consist of nucleotide / nucleoside analogs including deoxynucleotides / nucleosides. The overhang may be on the sense strand, the antisense strand, or any combination thereof. Further, the nucleotides of the overhang may be present on the 5'-end, 3'-end, or both ends of either the antisense strand or the sense strand of the dsRNA.

[0147] The dsRNA can be synthesized, for example, by use of an automated DNA synthesizer (such as those commercially available from Biosearch, Applied Biosystems, Inc., etc.) by standard methods known in the art as further described hereinafter.

[0148] The iRNA compounds of the present invention can be prepared using a two-step procedure. First, the individual strands of the double-stranded RNA molecule are prepared separately. Next, the component strands are annealed. The individual strands of the siRNA compounds can be prepared using solution-phase or solid-phase organic synthesis or both. Organic synthesis offers the advantage that oligonucleotide strands containing non-natural or modified nucleotides can be readily prepared. The single-stranded oligonucleotides of the present invention can be prepared using solution-phase or solid-phase organic synthesis or both.

[0149] In one aspect, the dsRNA of the present invention includes at least two nucleotide sequences, a sense sequence and an antisense sequence. The sense strand is selected from the group of sequences shown in any one of Tables 3, 4, 5, 6, 18, 19, 20, 21, and 23, and the corresponding antisense strand of the sense strand is selected from the group of sequences of any one of Tables 3, 4, 5, 6, 18, 19, 20, 21, and 23. In this aspect, one of the two sequences is complementary to the other of the two sequences, and one of the sequences is substantially complementary to the sequence of the mRNA generated during the expression of the C5 gene. Thus, in this aspect, the dsRNA will comprise two oligonucleotides, where one oligonucleotide is represented as the sense strand in any one of Tables 3, 4, 5, 6, 18, 19, 20, 21, and 23, and the second oligonucleotide is represented as the corresponding antisense strand of the sense strand in any one of Tables 3, 4, 5, 6, 18, 19, 20, 21, and 23. In one embodiment, the substantially complementary sequences of the dsRNA are contained in separate oligonucleotides. In another embodiment, the substantially complementary sequences of the dsRNA are contained in one oligonucleotide.

[0150] Some of the sequences in Tables 3, 4, 5, 6, 18, 19, 20, 21, and 23 are represented as modified and / or conjugate sequences, but it will be understood that the RNA of the iRNA of the present invention, for example, the dsRNA of the present invention, may include any one of the sequences described in Tables 3, 4, 5, 6, 18, 19, 20, 21, and 23 that are unmodified, unconjugated, and / or modified and / or conjugated, different from those described in the tables.

[0151] Those skilled in the art are well aware that dsRNA having a double-stranded structure of about 20 to 23 base pairs, for example, 21 base pairs, has been found to be particularly effective in inducing RNA interference (Elbashir et al., EMBO 2001, 20: 6877-6888). However, other those skilled in the art have found that shorter or longer RNA double-stranded structures may also be effective (Chu and Rana (2007) RNA 14: 1714-1719; Kim et al. (2005) Nat Biotech 23: 222-226). In the above embodiments, depending on the nature of the oligonucleotide sequences shown in any one of Tables 3, 4, 5, 6, 18, 19, 20, 21, and 23, the dsRNA described herein may include at least one strand with a minimum length of 21 nucleotides. It can be reasonably predicted that shorter double-strands having one of the sequences of any one of Tables 3, 4, 5, 6, 18, 19, 20, 21, and 23, with a few nucleotides subtracted at one or both ends, may be equally effective compared to the above dsRNA. Therefore, dsRNA having at least 15, 16, 17, 18, 19, 20, or more consecutive nucleotide sequences from one of the sequences of any one of Tables 3, 4, 5, 6, 18, 19, 20, 21, and 23, and whose ability to inhibit the expression of the C5 gene differs from that of the dsRNA containing the complete sequence by only about 5, 10, 15, 20, 25, or 30% or less inhibition, is considered to be within the scope of the present invention.

[0152] Furthermore, the RNA shown in any one of Tables 3, 4, 5, 6, 18, 19, 20, 21, and 23 identifies a site in the C5 transcript that is susceptible to cleavage via RISC. Accordingly, the invention further encompasses iRNAs that target within one of these sites. As used herein, an iRNA is said to target within a particular site of an RNA transcript if the iRNA promotes cleavage of the transcript at any location within that particular site. Such iRNAs generally will comprise at least about 15 contiguous nucleotides from one of the sequences shown in any one of Tables 3, 4, 5, 6, 18, 19, 20, 21, and 23, which are bound to an additional nucleotide sequence taken from a region adjacent to a selected sequence in the C5 gene.

[0153] Target arrays are generally about 15 to 30 nucleotides in length, but for any given target RNA cleavage, the suitability of specific sequences within this range varies. The various software packages and guidelines described herein provide guidance for identifying optimal target sequences for any given gene target, but a "window" or "mask" of a given size (as a non-limiting example, 21 nucleotides) is placed literally or metaphorically (including in silico) on the target RNA sequence to identify sequences within the size range that can function as target sequences. An empirical approach can also be taken. The next potential target sequence can be identified by gradually moving the sequence "window" 1 nucleotide upstream or downstream of the initial target sequence position until a complete set of possible sequences is identified for any given target size selected. This process can identify the RNA sequence that mediates the best inhibition of target gene expression when targeted with an iRNA agent, along with the systematic synthesis and testing of the identified sequences to identify the sequences that function optimally (using assays described herein or known in the art). Thus, for example, the sequences identified in any one of Tables 3, 4, 5, 6, 18, 19, 20, 21, and 23 represent effective target sequences, but further optimization of inhibition efficiency can be achieved, it is believed, by gradually "moving the window" 1 nucleotide upstream or downstream of a given sequence to identify sequences with equivalent or better inhibitory properties.

[0154] Furthermore, for any array identified in any one of Tables 3, 4, 5, 6, 18, 19, 20, 21, and 23, additional optimization can be achieved by systematically adding or removing nucleotides to generate longer or shorter arrays, and testing the arrays generated by moving a window of longer or shorter size above or below the target RNA from that point. Also, in the inhibition assays known in the art and / or described herein, by coupling this approach for generating new candidate targets with the testing of the efficacy of iRNAs based on their target sequences, the efficiency of inhibition can be further improved. Additionally, such optimized arrays can be adjusted, for example, by introducing modified nucleotides as described herein or known in the art, adding or changing overhangs, or other modifications known in the art and / or described herein for further optimizing the molecule as an expression inhibitor (e.g., increasing serum stability or circulatory half-life, increasing thermal stability, improving membrane permeation delivery, targeting to specific locations or cell types, increasing interaction with silencing pathway enzymes, increasing release from endosomes).

[0155] The iRNAs described herein may contain one or more mismatches with the target sequence. In one embodiment, the iRNAs described herein contain three or fewer mismatches. When the antisense strand of the iRNA contains a mismatch with the target sequence, it is preferred that the region of the mismatch is not located at the center of the complementary region. When the antisense strand of the iRNA contains a mismatch with the target sequence, the mismatch is preferably restricted within the last 5 nucleotides from either the 5' or 3' end of the complementary region. For example, for a 23-nucleotide iRNA agent, the strand complementary to the region of the C5 gene generally contains no mismatches within the central 13 nucleotides. It is possible to determine whether an iRNA containing a mismatch with the target sequence is effective in inhibiting the expression of the C5 gene using the methods described herein or methods known in the art. Considering the effectiveness of an iRNA with a mismatch in inhibiting the expression of the C5 gene is particularly important when it is known that a specific complementary region in the C5 gene has polymorphic sequence variations within the population.

[0156] III. Modified iRNAs of the Invention In one embodiment, the RNA of the iRNA of the invention, for example, dsRNA, is not modified, for example, does not contain chemical modifications and / or conjugates known in the art and described herein. In another embodiment, the RNA of the iRNA of the invention, for example, dsRNA, is chemically modified to improve stability or other beneficial properties. In certain embodiments of the invention, substantially all of the nucleotides of the iRNA of the invention are modified. In other embodiments of the invention, all of the nucleotides of the iRNA of the invention are modified. The iRNA of the invention in which "substantially all of the nucleotides are modified" is mostly modified but not completely modified and may contain 5 or fewer, 4 or fewer, 3 or fewer, 2 or fewer, or 1 or fewer unmodified nucleotides.

[0157] The nucleic acids employed in the present invention can be synthesized and / or modified by methods well established in the art, such as those described in “Current protocols in nucleic acid chemistry,” Beaucage, S.L. et al. (Eds.), John Wiley & Sons, Inc., New York, NY, USA, which is hereby incorporated by reference. Modifications include, for example, terminal modifications, such as 5′-terminal modifications (phosphorylation, conjugation, inverted linkage) or 3′-terminal modifications (conjugation, DNA nucleotides, inverted linkage, etc.); base modifications, such as substitution of a base with a stable base, an unstable base, or a base that base pairs with a wide range of partners, removal of a base (non-basic nucleotide), or a conjugated base; sugar modifications (e.g., at the 2′ or 4′ position) or substitution of the sugar; and / or backbone modifications, including modification or substitution of the phosphodiester bond. Specific examples of iRNA compounds useful in the embodiments described herein include, but are not limited to, RNAs that contain a modified backbone or do not contain natural internucleoside linkages. RNAs having a modified backbone include, in particular, those that do not have a phosphorus atom in the backbone. For the purposes of this specification and as sometimes referred to in the art, a modified RNA that does not have a phosphorus atom in the internucleoside backbone can also be considered an oligonucleoside. In certain embodiments, the modified iRNA has a phosphorus atom in its internucleoside backbone.

[0158] Examples of modified RNA backbones include phosphorothioate, chiral phosphorothioate, phosphorodithioate, phosphotriester, aminoalkyl phosphotriester, methylphosphonate, and other alkylphosphonates including 3'-alkylene phosphonate and chiral phosphonate, phosphinate, phosphoramidate including 3'-aminophosphoramidate and aminoalkylphosphoramidate, thionophosphoramidate, thionoalkylphosphonate, thionoalkylphosphotriester, and boranophosphate having a normal 3'-5' linkage, their 2'-5' linkage analogs, and those having an inverted polarity where adjacent pairs of nucleoside units are linked 3'-5' to 5'-3' or 2'-5' to 5'-2'. Also included are various salts, mixed salts, and free acid forms.

[0159] Representative U.S. patents that teach the preparation of the above phosphorus-containing linkages include, but are not limited to, U.S. Patent No. 3,687,808; No. 4,469,863; No. 4,476,301; No. 5,023,243; No. 5,177,195; No. 5,188,897; No. 5,264,423; No. 5,276,019; No. 5,278,302; No. 5,286,717; No. 5,321,131; No. 5,399,676; No. 5,405,939; No. 5,453,496; No. 5,455,233; No. 5,466,677; No. 5,476,925; No. 5,519,126; No. 5,536,821; No. 5,541,316; No. 5,550,111; No. 5,563,253; No. 5,571,799; No. 5,587,361; No. 5,625,050; No. 6,028,188; No. 6,124,445; No. 6,160,109; No. 6,169,170; No. 6,172,209; No. 6,239,265; No. 6,277,603; No. 6,326,199; No. 6,346,614; No. 6,444,423; No. 6,531,590; No. 6,534,639; No. 6,608,035; No. 6,683,167; No. 6,858,715; No. 6,867,294; No. 6,878,805; No. 7,015,315; No. 7,041,816; No. 7,273,933; No. 7,321,029; and U.S. Reissue Patent No. RE39464, the entire contents of each of which are hereby incorporated by reference into this specification.

[0160] Modified RNA backbones that do not contain phosphorus atoms within have a backbone formed by short-chain alkyl or cycloalkyl nucleoside linkages, mixed heteroatom and alkyl or cycloalkyl nucleoside linkages, or one or more short-chain heteroatom or heterocyclic nucleoside linkages. These include those having a morpholino linkage (partially formed from the sugar moiety of the nucleoside); a siloxane backbone; sulfide, sulfoxide, and sulfone backbones; formacetyl and thioformacetyl backbones; methyleneformacetyl and thioformacetyl backbones; alkene-containing backbones; sulfamate backbones; methyleneimino and methylenehydrazino backbones; sulfonate and sulfonamide backbones; amide backbones; and other ones having a mixed N, O, S, and CH 2 and other ones having component parts.

[0161] Representative U.S. patents that teach the preparation of the above oligonucleosides include, but are not limited to, U.S. Patent Nos. 5,034,506; 5,166,315; 5,185,444; 5,214,134; 5,216,141; 5,235,033; 5,264,562; 5,264,564; 5,405,938; 5,434,257; 5,466,677; 5,470,967; 5,489,677; 5,541,307; 5,561,225; 5,596,086; 5,602,240; 5,608,046; 5,610,289; 5,618,704; 5,623,070; 5,663,312; 5,633,360; 5,677,437; and 5,677,439, the entire contents of each of which are hereby incorporated by reference into this specification.

[0162] In other embodiments, suitable RNA mimics are contemplated for use in iRNA, where both the sugar and the internucleoside linkage, i.e., the backbone of the nucleotide unit, are replaced with novel groups. The base units are maintained for hybridization with a suitable nucleic acid target compound. One such oligomeric compound that has been shown to have excellent hybridization properties is an RNA mimic called peptide nucleic acid (PNA). In PNA compounds, the sugar backbone of RNA is replaced with an amide-containing backbone, specifically, an aminoethylglycine backbone. The nucleobases are retained and are attached directly or indirectly to the azanitrogen atoms of the amide portions of the backbone. Representative U.S. patents that teach the preparation of PNA compounds include, but are not limited to, U.S. Patent Nos. 5,539,082; 5,714,331; and 5,719,262, the entire contents of each of which are incorporated herein by reference. Further PNA compounds suitable for use in the iRNA of the present invention are described, for example, in Nielsen et al., Science, 1991, 254, 1497-1500.

[0163] One embodiment taken up in the present invention includes RNAs having phosphorothioate backbones and oligonucleosides having heteroatom backbones, particularly the --CH of U.S. Patent No. 5,489,677 described above. 2 --NH--CH 2 -、--CH 2 --N(CH 3 )--O--CH 2 --[known as the methylene(methylimino) or MMI backbone], --CH 2 --O--N(CH 3 )--CH 2 --、--CH 2 --N(CH 3 )--N(CH 3 )--CH 2 --and --N(CH 3 )--CH 2 --CH 2 --[wherein the natural phosphodiester backbone is --O--P--O--CH 2--], and the amide backbone of the aforementioned U.S. Patent No. 5,602,240. In some embodiments, the RNA featured herein has the morpholino backbone structure of the aforementioned U.S. Patent No. 5,034,506.

[0164] Modified RNAs may also contain one or more substituted sugar moieties. The iRNAs, e.g., dsRNAs, featured herein, may include one of the following at the 2' position: OH; F; O-, S-, or N-alkyl; O-, S-, or N-alkenyl; O-, S-, or N-alkynyl; or O-alkyl-O-alkyl, where alkyl, alkenyl, and alkynyl are substituted or unsubstituted C 1 ~C 10 Alkyl or C 2 ~C 10 It can be alkenyl and alkynyl. Exemplary suitable modifications include O[(CH 2 ) n O] m CH 3 , O(CH 2 ). n OCH 3 , O(CH 2 ) n NH 2 , O(CH 2 ) n CH 3 , O(CH 2 ) n O.N.H. 2 , and O(CH 2 ) n ON[(CH 2 ) n CH 3 )] 2 wherein n and m are from 1 to about 10. In other embodiments, the dsRNA comprises at the 2' position: 1 ~C 10 Lower alkyl, substituted lower alkyl, alkaryl, aralkyl, O-alkaryl or O-aralkyl, SH, SCH 3 , OCN, Cl, Br, CN, CF 3 , OCF 3 , SOCH 3 , S.O. 2 CH3 、ONO 2 、NO 2 、N 3 、NH 2 、heterocycloalkyl, heterocycloalkaryl, aminoalkylamino, polyalkylamino, substituted silyl, RNA cleavage group, reporter group, intercalator, a group that improves the pharmacodynamic properties of iRNA, or a group that improves the pharmacokinetic properties of iRNA, and one of other substituents having similar properties. In certain embodiments, the modification is 2'-methoxyethoxy (also known as 2'-O-(2-methoxyethyl) or 2'-MOE), i.e., 2'-O--CH 2 CH 2 OCH 3 )(Martin et al., Helv. Chim. Acta, 1995, 78:486-504), i.e., an alkoxy-alkoxy group. Another exemplary modification is 2'-dimethylaminooxyethoxy, also known as 2'-DMAOE, described in the examples hereinbelow, i.e., O(CH 2 ) 2 ON(CH 3 ) 2 group, and 2'-dimethylaminoethoxyethoxy (also known in the art as 2'-O-dimethylaminoethylethyl or 2'-DMAEOE), i.e., 2'-O--CH 2 --O--CH 2 --N(CH 2 ) 2 .

[0165] Other modifications are 2'-methoxy (2'-OCH 3 ), 2'-aminopropoxy (2'-OCH 2 CH 2 CH 2 NH 2) and 2'-fluoro (2'-F). Similar modifications can also be made at other positions in the RNA of the iRNA, particularly at the 3'-terminal nucleotide or at the 3'-position of the sugar in the 2'-5'-linked dsRNA and at the 5'-position of the 5'-terminal nucleotide. The iRNA can also have a sugar mimic such as a cyclobutyl moiety instead of a pentofuranosyl sugar. Representative U.S. patents teaching the preparation of such modified sugar structures include, but are not limited to, U.S. Patent Nos. 4,981,957; 5,118,800; 5,319,080; 5,359,044; 5,393,878; 5,446,137; 5,466,786; 5,514,785; 5,519,134; 5,567,811; 5,576,427; 5,591,722; 5,597,909; 5,610,300; 5,627,053; 5,639,873; 5,646,265; 5,658,873; 5,670,633; and 5,700,920, some of which have the same owner as the present application. The entire contents of each of the above are hereby incorporated by reference into this specification.

[0166] iRNAs can also include nucleobase (often simply referred to as "base" in the art) modifications or substitutions. As used herein, "unmodified" or "natural" nucleobases include the purine bases adenine (A) and guanine (G), and the pyrimidine bases thymine (T), cytosine (C), and uracil (U). Modified nucleobases include deoxy-thymine (dT), 5-methylcytosine (5-me-C), 5-hydroxymethylcytosine, xanthine, hypoxanthine, 2-aminoadenine, 6-methyl and other alkyl derivatives of adenine and guanine, 2-propyl and other alkyl derivatives of adenine and guanine, 2-thiouracil, 2-thiothymine and 2-thiocytosine, 5-halouracil and cytosine, 5-propynyluracil and cytosine, 6-azauracil, cytosine and thymine, 5-uracil (pseudouracil), 4-thiouracil, 8-halo, 8-amino, 8-thiol, 8-thioalkyl, 8-hydroxyl and other 8-substituted adenines and guanines, 5-halo, particularly 5-bromo, 5-trifluoromethyl and other 5-substituted uracils and cytosines, 7-methylguanine and 7-methyladenine, 8-azaguanine and 8-azaadenine, 7-deazaguanine and 7-deazaadenine (daazaadenine) and other synthetic and natural nucleobases such as 3-deazaguanine and 3-deazaadenine.Additional nucleobases include those disclosed in U.S. Patent No. 3,687,808; those disclosed in Modified Nucleosides in Biochemistry, Biotechnology and Medicine, Herdewijn, P. ed. Wiley-VCH, 2008; those disclosed in Concise Encyclopedia Of Polymer Science and Engineering, pp. 858-859, Kroschwitz, J.L, ed. John Wiley & Sons, 1990; those disclosed by Englisch et al., Angewandte Chemie, International Edition, 1991, 30, 613; and those disclosed by Sanghvi, Y S., Chapter 15, dsRNA Research and Applications, pp. 289-302, Crooke, S.T. and Lebleu, B., Ed., CRC Press, 1993. Some of these nucleobases are particularly useful for enhancing the binding affinity of the oligomeric compounds taken up in the present invention. These include 5-substituted pyrimidines, 6-azapyrimidines and N-2, N-6 and 0-6 substituted purines, including 2-aminopropyladenine, 5-propynyluracil and 5-propynylcytosine. The 5-methylcytosine substituent has been shown to increase nucleic acid duplex stability by 0.6 to 1.2 °C (Sanghvi, Y.S., Crooke, S.T. and Lebleu, B., Eds., dsRNA Research and Applications, CRC Press, Boca Raton, 1993, pp. 276-278) and is an exemplary base substitution, especially when combined with 2'-O-methoxyethyl sugar modifications.

[0167] Representative U.S. patents that teach the preparation of some of the above-described modified nucleobases and other modified nucleobases include, but are not limited to, the above-mentioned U.S. Patent Nos. 3,687,808; 4,845,205; 5,130,30; 5,134,066; 5,175,273; 5,367,066; 5,432,272; 5,457,187; 5,459,255; 5,484,908; 5,502,177; 5,525,711; 5,552,540; 5,587,469; 5,594,121; 5,596,091; 5,614,617; 5,681,941; 5,750,692; 6,015,886; 6,147,200; 6,166,197; 6,222,025; 6,235,887; 6,380,368; 6,528,640; 6,639,062; 6,617,438; 7,045,610; 7,427,672; and 7,495,088, the entire contents of each of which are hereby incorporated by reference.

[0168] The RNA of the iRNA can also be modified to include one or more bicyclic sugar moieties. A "bicyclic sugar" is a furanosyl ring modified by a bridge of two atoms. A "bicyclic nucleoside" ("BNA") is a nucleoside having a sugar moiety that includes a bridge that joins two carbon atoms of the sugar ring, thereby forming a bicyclic ring system. In certain embodiments, the bridge joins the 4'-carbon and the 2'-carbon of the sugar ring. Thus, in one embodiment, the agent of the invention may include an RNA of an iRNA that is similarly modified to include one or more locked nucleic acids (LNAs). A locked nucleic acid is a nucleotide having a modified ribose moiety, the ribose moiety including an additional bridge that joins the 2' and 4' carbons. In other words, an LNA is a nucleotide that includes a bicyclic sugar moiety that includes a 4'-CH2-O-2' bridge. This structure effectively "locks" the ribose in the 3'-endo conformational configuration. Addition of locked nucleic acids to siRNA has been shown to increase siRNA stability in serum and decrease off-target effects (Elmen, J. et al., (2005) Nucleic Acids Research 33(1):439-447; Mook, O.R. et al., (2007) Mol Canc Ther 6(3):833-843; Grunweller, A. et al., (2003) Nucleic Acids Research 31(12):3185-3193).

[0169] Examples of bicyclic nucleosides for use in the polynucleotides of the present invention include, but are not limited to, nucleosides containing a bridge between the 4' and 2' ribosyl ring atoms. In certain embodiments, the antisense polynucleotide agents of the present invention comprise one or more bicyclic nucleosides containing a 4'-2' bridge. Examples of such 4'-2' bridged bicyclic nucleosides include, but are not limited to, 4'-(CH2)-O-2' (LNA); 4'-(CH2)-S-2'; 4'-(CH2)2-O-2' (ENA); 4'-CH(CH3)-O-2' (also referred to as "constrained ethyl" or "cEt") and 4'-CH(CH2OCH3)-O-2' (and its analogs; see, e.g., U.S. Patent No. 7,399,845); 4'-C(CH3)(CH3)-O-2' (and its analogs; see, e.g., U.S. Patent No. 8,278,283); 4'-CH2-N(OCH3)-2' (and its analogs; see, e.g., U.S. Patent No. 8,278,425); 4'-CH2-O-N(CH3)-2' (see, e.g., U.S. Patent Application Publication No. 2004 / 0171570); 4'-CH2-N(R)-O-2' where R is H, C1-C12 alkyl, or a protecting group (see, e.g., U.S. Patent No. 7,427,672); 4'-CH2-C(H)(CH3)-2' (see, e.g., Chattopadhyaya et al., J. Org. Chem., 2009, 74, 118-134); and 4'-CH2-C(=CH2)-2' (and its analogs; see, e.g., U.S. Patent No. 8,278,426). The entire contents of each of the above are hereby incorporated by reference into this specification.

[0170] Additional representative U.S. patents and U.S. patent publications that teach the preparation of locked nucleic acid nucleotides include, but are not limited to, U.S. Patent No. 6,268,490; 6,525,191; 6,670,461; 6,770,748; 6,794,499; 6,998,484; 7,053,207; 7,034,133; 7,084,125; 7,399,845; 7,427,672; 7,569,686; 7,741,457; 8,022,193; 8,030,467; 8,278,425; 8,278,426; 8,278,283; U.S. Patent Application Publication No. 2008 / 0039618; and U.S. Patent Application Publication No. 2009 / 0012281, the entire contents of each of which are hereby incorporated by reference.

[0171] Any of the above bicyclic nucleosides can be prepared having one or more stereochemical sugar configurations including, for example, α-L-ribofuranose and β-D-ribofuranose (see WO 99 / 14226).

[0172] The RNA of the iRNA can also be modified to include one or more constrained ethyl nucleotides. As used herein, "constrained ethyl nucleotide" or "cEt" is a locked nucleic acid containing a bicyclic sugar moiety including a 4'-CH(CH3)-O-2' bridge. In one embodiment, the constrained ethyl nucleotide is in the S conformation, referred to herein as "S-cEt".

[0173] The iRNA of the present invention may also include one or more "conformationally restricted nucleotides" ("CRNs"). A CRN is a nucleotide analog having a linker that binds the C2' and C4' carbons of ribose or the C3 and C5' carbons of ribose. The CRN locks the ribose ring into a stable conformation and increases the hybridization affinity for mRNA. The linker is of sufficient length to position oxygen optimally for stability and affinity so as to reduce ribose ring packing.

[0174] Representative publications teaching some preparations of the above CRNs include, but are not limited to, U.S. Patent Application Publication No. 2013 / 0190383; and PCT Publication WO 2013 / 036868, the entire contents of each of which are incorporated herein by reference.

[0175] One or more of the nucleotides of the iRNA of the present invention may also include hydroxymethyl-substituted nucleotides. A "hydroxymethyl-substituted nucleotide" is an acyclic 2'-3'-seco-nucleotide, also referred to as a "non-locked nucleic acid" ("UNA") modification.

[0176] Representative U.S. patent publications teaching the preparation of UNA include, but are not limited to, U.S. Patent No. 8,314,227; and U.S. Patent Application Publication Nos. 2013 / 0096289; 2013 / 0011922; and 2011 / 0313020, the entire contents of each of which are incorporated herein by reference.

[0177] Potentially stable modifications to the ends of RNA molecules can include N-(acetylaminocaproyl)-4-hydroxyprolinol (Hyp-C6-NHAc), N-(caproyl4-hydroxyprolinol (Hyp-C6), N-(acetyl-4-hydroxyprolinol (Hyp-NHAc), thymidine-2'-O-deoxythymidine (ether), N-(aminocaproyl)-4-hydroxyprolinol (Hyp-C6-amino), 2-docosanoyl-uridine-3”-phosphate, inverted base dT (idT), and the like. The disclosure of this modification can be found in PCT Publication No. WO 2011 / 005861 pamphlet.

[0178] A. Modified iRNA containing the motif of the present invention In certain embodiments of the present invention, double-stranded RNAi agents of the present invention include, for example, agents having chemical modifications disclosed in U.S. Provisional Patent Application No. 61 / 561,710, filed November 18, 2011, or PCT Publication No. WO 2010 / 065691, filed November 16, 2012, the entire contents of each of which are hereby incorporated by reference in their entirety.

[0179] As shown herein and in U.S. Provisional Patent Application No. 61 / 561,710 or PCT Application No. PCT / US2010 / 065691, better results are obtained by introducing one or more motifs of three identical modifications on three consecutive nucleotides into the sense strand and / or the antisense strand of the RNAi agent, particularly at or near the cleavage site. In certain embodiments, the sense strand and the antisense strand of the RNAi agent can alternatively be fully modified. The introduction of these motifs disrupts the modification pattern of the sense strand and / or the antisense strand, if present. The RNAi agent can be optionally conjugated, for example, on the sense strand, with a GalNAc derivative ligand. The resulting RNAi agent exhibits better gene silencing activity.

[0180] More specifically, it has been unexpectedly discovered that when the sense strand and the antisense strand of a double-stranded RNAi agent are fully modified such that they have one or more motifs of three identical modifications on three consecutive nucleotides at or near the cleavage site of at least one strand of the RNAi agent, the gene silencing activity of the RNAi agent is significantly improved.

[0181] Accordingly, the present invention provides a double-stranded RNAi agent capable of inhibiting the expression of a target gene (i.e., the complement component C5 (C5) gene) in vivo. The RNAi agent includes a sense strand and an antisense strand. Each strand of the RNAi agent can be in the range of 12 to 30 nucleotides in length. For example, each strand can be 14 to 30 nucleotides in length, 17 to 30 nucleotides in length, 25 to 30 nucleotides in length, 27 to 30 nucleotides in length, 17 to 23 nucleotides in length, 17 to 21 nucleotides in length, 17 to 19 nucleotides in length, 19 to 25 nucleotides in length, 19 to 23 nucleotides in length, 19 to 21 nucleotides in length, 21 to 25 nucleotides in length, or 21 to 23 nucleotides in length.

[0182] The sense strand and the antisense strand typically form a double-stranded RNA ("dsRNA"), also referred to herein as an "RNAi agent". The double-stranded region of the RNAi agent can be 12 to 30 nucleotide pairs in length. For example, the double-stranded region can be 14 to 30 nucleotide pairs in length, 17 to 30 nucleotide pairs in length, 27 to 30 nucleotide pairs in length, 17 to 23 nucleotide pairs in length, 17 to 21 nucleotide pairs in length, 17 to 19 nucleotide pairs in length, 19 to 25 nucleotide pairs in length, 19 to 23 nucleotide pairs in length, 19 to 21 nucleotide pairs in length, 21 to 25 nucleotide pairs in length, or 21 to 23 nucleotide pairs in length. In another example, the double-stranded region is selected from 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, and 27 nucleotides in length.

[0183] In one embodiment, the RNAi agent may include one or more overhang regions and / or capping groups at the 3'-end, 5'-end, or both ends of one or both strands. The overhang may be 1 to 6 nucleotides in length, for example, 2 to 6 nucleotides in length, 1 to 5 nucleotides in length, 2 to 5 nucleotides in length, 1 to 4 nucleotides in length, 2 to 4 nucleotides in length, 1 to 3 nucleotides in length, 2 to 3 nucleotides in length, or 1 to 2 nucleotides in length. The overhang may be the result of one strand being longer than the other, or the result of two strands of the same length being staggered. The overhang may form a mismatch with the target mRNA, or the overhang may be complementary to the target gene sequence, or it may be another sequence. The first and second strands may also be joined by additional bases, for example, to form a hairpin, or by other non-base linkers.

[0184] In one embodiment, each nucleotide in the overhang region of the RNAi agent may independently be a modified or unmodified nucleotide including a 2'-sugar modification such as, but not limited to, 2'-F, 2'-O-methyl, thymidine (T), 2'-O-methoxyethyl-5-methyluridine (Teo), 2'-O-methoxyethyladenosine (Aeo), 2'-O-methoxyethyl-5-methylcytidine (m5Ceo), and any combination thereof. For example, TT may be an overhang sequence for either end on either strand. The overhang may form a mismatch with the target mRNA, or the overhang may be complementary to the target gene sequence, or it may be another sequence.

[0185] The 5'- or 3'-overhangs in the sense strand, antisense strand, or both strands of the RNAi agent can be phosphorylated. In certain embodiments, the overhang region comprises two nucleotides having a phosphorothioate between two nucleotides, where the two nucleotides can be the same or different. In one embodiment, the overhang is present at the 3'-end of the sense strand, antisense strand, or both strands. In one embodiment, this 3'-overhang is present in the antisense strand. In one embodiment, this 3'-overhang is present in the sense strand.

[0186] The RNAi agent can contain only one overhang that can enhance the interference activity of RNAi without affecting its overall stability. For example, a single-stranded overhang can be located at the 3'-end of the sense strand or the 3'-end of the antisense strand. RNAi can also have blunt ends located at the 5'-end of the antisense strand (or the 3'-end of the sense strand) or vice versa. Generally, the antisense strand of RNAi has a nucleotide overhang at the 3'-end and a blunt 5'-end. Without wishing to be bound by theory, the asymmetric blunt ends at the 5'-end and the 3'-end overhang of the antisense strand favorably act on the introduction of the guide strand into the RISC process.

[0187] In one embodiment, the RNAi agent is a 19-nucleotide-long double-ended bluntmer, and the sense strand contains at least one motif of three 2'-F modifications in three consecutive nucleotides at positions 7, 8, and 9 from the 5'-end. The antisense strand contains at least one motif of three 2'-O-methyl modifications in three consecutive nucleotides at positions 11, 12, and 13 from the 5'-end.

[0188] In another embodiment, the RNAi agent is a blunt-ended double strand 20 nucleotides in length, and the sense strand contains at least one motif of three 2'-F modifications in three consecutive nucleotides at positions 8, 9, and 10 from the 5' end. The antisense strand contains at least one motif of three 2'-O-methyl modifications in three consecutive nucleotides at positions 11, 12, and 13 from the 5' end.

[0189] In yet another embodiment, the RNAi agent is a blunt-ended double strand 21 nucleotides in length, and the sense strand contains at least one motif of three 2'-F modifications in three consecutive nucleotides at positions 9, 10, and 11 from the 5' end. The antisense strand contains at least one motif of three 2'-O-methyl modifications in three consecutive nucleotides at positions 11, 12, and 13 from the 5' end.

[0190] In one embodiment, the RNAi agent comprises a 21-nucleotide sense strand and a 23-nucleotide antisense strand. The sense strand contains at least one motif of three 2'-F modifications in three consecutive nucleotides at positions 9, 10, and 11 from the 5'-end; the antisense strand contains at least one motif of three 2'-O-methyl modifications in three consecutive nucleotides at positions 11, 12, and 13 from the 5'-end. One end of the RNAi agent is blunt, while the other end contains a two-nucleotide overhang. Preferably, the two-nucleotide overhang is at the 3'-end of the antisense strand. When the two-nucleotide overhang is at the 3'-end of the antisense strand, there may be two phosphorothioate internucleotide linkages between the three terminal nucleotides, two of the three nucleotides being overhang nucleotides and the third nucleotide being the paired nucleotide adjacent to the overhang nucleotide. In one embodiment, the RNAi agent further has two phosphorothioate internucleotide linkages between the three terminal nucleotides at both the 5'-end of the sense strand and the 5'-end of the antisense strand. In one embodiment, all nucleotides in the sense and antisense strands of the RNAi agent, including those nucleotides that are part of the motif, are modified nucleotides. In one embodiment, each residue is independently modified, for example, with 2'-O-methyl or 3'-fluoro in an alternating motif. Optionally, the RNAi agent further comprises a ligand (preferably, GalNAc 3 ).

[0191] In one embodiment, the RNAi agent includes a sense strand and an antisense strand. The sense strand is 25 to 30 nucleotide residues in length. Starting from the 5'-terminal nucleotide (position 1), positions 1 to 23 of the first strand contain at least 8 ribonucleotides. The antisense strand is 36 to 66 nucleotide residues in length. Starting from the 3'-terminal nucleotide, it contains at least 8 ribonucleotides at the position paired with positions 1 to 23 of the sense strand to form a double strand. At least the 3'-terminal nucleotides of the antisense strand are not paired with the sense strand, and a maximum of 6 consecutive 3'-terminal nucleotides are not paired with the sense strand, thereby forming a 3'-single-stranded overhang of 1 to 6 nucleotides. The 5'-terminal of the antisense strand contains 10 to 30 consecutive nucleotides not paired with the sense strand, thereby forming a single-stranded 5'-overhang of 10 to 30 nucleotides. At least the 5'-terminal and 3'-terminal nucleotides of the sense strand are bases paired with the nucleotides of the antisense strand when the sense strand and the antisense strand are aligned for maximum complementarity, thereby forming a substantially double-stranded region between the sense strand and the antisense strand. The antisense strand is sufficiently complementary to the target RNA along at least 19 ribonucleotides of the antisense strand length so as to reduce the expression of the target gene when the double-stranded nucleic acid is introduced into mammalian cells. The sense strand contains at least one motif of three 2'-F modifications in three consecutive nucleotides, where at least one of the motifs is present at or near the cleavage site. The antisense strand contains at least one motif of three 2'-O-methyl modifications in three consecutive nucleotides at or near the cleavage site.

[0192] In one embodiment, the RNAi agent comprises a sense strand and an antisense strand. The RNAi agent comprises a first strand having a nucleotide length of at least 25 and 29 or less, and a second strand having a length of 30 nucleotides or less, which comprises at least one motif of three 2'-O-methyl modifications in three consecutive nucleotides at positions 11, 12, and 13 from the 5' end; the 3' end of the first strand and the 5' end of the second strand form blunt ends, the second strand is 1 to 4 nucleotides longer than the first strand at its 3' end, the double-stranded region is at least 25 nucleotides long, and the second strand is sufficiently complementary to the target mRNA along at least 19 nucleotides of the second strand length so that when the RNAi agent is introduced into mammalian cells, it reduces the expression of the target gene. Dicer cleavage of the RNAi agent preferentially yields siRNAs containing the 3' end of the second strand, thereby reducing the expression of the target gene in mammals. Optionally, the RNAi agent further comprises a ligand.

[0193] In one embodiment, the sense strand of the RNAi agent comprises at least one motif of three identical modifications in three consecutive nucleotides, and one of the motifs is present at the cleavage site of the sense strand.

[0194] In one embodiment, the antisense strand of the RNAi agent can also comprise at least one motif of three identical modifications in three consecutive nucleotides, and one of the motifs is present at or near the cleavage site of the antisense strand.

[0195] In an RNAi agent having a double-stranded region with a length of 17 to 23 nucleotides, the cleavage site of the antisense strand is typically near the 10th, 11th, and 12th positions from the 5' end. Therefore, three identical modified motifs can start counting from the first nucleotide from the 5' end of the antisense strand, or from the first paired nucleotide within the double-stranded region from the 5' end of the antisense strand, and can be present at the 9th, 10th, 11th positions; 10th, 11th, 12th positions; 11th, 12th, 13th positions; 12th, 13th, 14th positions; or 13th, 14th, 15th positions of the antisense strand. The cleavage site in the antisense strand can also vary depending on the length of the double-stranded region of the RNAi from the 5' end.

[0196] The sense strand of the RNAi agent may contain at least one motif of three identical modifications in three consecutive nucleotides at the cleavage site of the strand; the antisense strand may have at least one motif of three identical modifications in three consecutive nucleotides at or near the cleavage site of the strand. When the sense strand and the antisense strand form a dsRNA duplex, the sense strand and the antisense strand can be aligned such that one motif of three nucleotides in the sense strand and one motif of three nucleotides in the antisense strand have at least one nucleotide overlap, that is, at least one of the three nucleotides of the motif in the sense strand forms a base pair with at least one of the three nucleotides of the motif in the antisense strand. Alternatively, at least two nucleotides may overlap, or all three nucleotides may overlap.

[0197] In one embodiment, the sense strand of the RNAi agent may include two or more motifs of three identical modifications in three consecutive nucleotides. The first motif may be present at or near the cleavage site of the strand, and the other motifs may be wing modifications. As used herein, the term "wing modification" refers to a motif present in another part of the strand away from the motif at or near the cleavage site of the same strand. The wing modification is adjacent to the first motif or separated by at least one or more nucleotides. When the motifs are directly adjacent to each other, the chemical structures of the motifs are different from each other. When the motifs are separated by one or more nucleotides, the chemical structures may be the same or different. Two or more wing modifications may be present. For example, when two wing modifications are present, each wing modification may be present at one end with respect to the first motif at or near the cleavage site or on either side of the lead motif.

[0198] Similar to the sense strand, the antisense strand of the RNAi agent may also include two or more motifs of three identical modifications in three consecutive nucleotides, and at least one of the motifs is present at or near the cleavage site of the strand. This antisense strand may also include one or more wing modifications in a sequence similar to the wing modifications that may be present in the sense strand.

[0199] In one embodiment, the wing modification in the sense strand or antisense strand of the RNAi agent typically does not include the first one or two terminal nucleotides at the 3' end, 5' end, or both ends of the strand.

[0200] In another embodiment, the wing modification in the sense strand or antisense strand of the RNAi agent typically does not include the first one or two paired nucleotides within the double-stranded region at the 3' end, 5' end, or both ends of the strand.

[0201] When the sense strand and the antisense strand of the RNAi agent each contain at least one wing modification, the wing modifications may be located at the same end of the double-stranded region and may have an overlap of one, two, or three nucleotides.

[0202] When the sense strand and the antisense strand of the RNAi agent each contain at least two wing modifications, the sense strand and the antisense strand may be such that two modifications from one strand are each located at one end of the double-stranded region and have an overlap of one, two, or three nucleotides; two modifications from one strand are each located at the other end of the double-stranded region and have an overlap of one, two, or three nucleotides; and two modifications from one strand are located on each side of the lead motif and may be aligned to have an overlap of one, two, or three nucleotides in the double-stranded region.

[0203] In one embodiment, all nucleotides in the sense strand and the antisense strand of the RNAi agent, including nucleotides that are part of the motif, may be modified. Each nucleotide may be modified with the same or different modifications, which may include one or more changes to one or both of the non-bridging phosphate oxygen and / or the bridging phosphate oxygen; changes to the ribose sugar component, such as changes to the 2'-hydroxyl of the ribose sugar; large-scale substitution of the phosphate moiety with a "dephospho" linker; modification or substitution of natural bases; and substitution or modification of the ribose-phosphate backbone.

[0204] Because nucleic acids are polymers of subunits, many modifications, such as bases, or modifications of the phosphate moiety, or non-bridging O of the phosphate moiety, are present at repeated positions within the nucleic acid. In some cases, the modification may be present at all positions of interest in the nucleic acid, but in many cases it is not. As an example, the modification may be present only at the 3' or 5' terminal positions, or only in the terminal region, such as a position on the terminal nucleotide or only the last 2, 3, 4, 5, or 10 nucleotides of the strand. The modification may be present in double-stranded regions, single-stranded regions, or both. The modification may be present only in the double-stranded region of RNA, or only in the single-stranded region of RNA. For example, phosphorothioate modification at the non-bridging O position may be present only at one or both ends, or only in the terminal region, such as a position on the terminal nucleotide or only the last 2, 3, 4, 5, or 10 nucleotides of the strand, or may be present in double-stranded and single-stranded regions, particularly at the ends. The 5' end or both ends may be phosphorylated.

[0205] For example, it may be possible to enhance stability, include specific bases in the overhang, or include modified nucleotides or nucleotide surrogates in single-stranded overhangs, such as 5' or 3' overhangs, or both. For example, it may be desirable to include purine nucleotides in the overhang. In certain embodiments, all or some of the bases in the 3' or 5' overhang may be modified, for example, with the modifications described herein. Modifications can include, for example, the use of modifications at the 2' position of the ribose sugar by modifications known in the art, such as the use of deoxyribonucleotides, 2'-deoxy-2'-fluoro (2'-F) or 2'-O-methyl modifications instead of the ribose sugar of the nucleobase, and modifications of the phosphate group, such as phosphorothioate modification. The overhang need not be homologous to the target sequence.

[0206] In one embodiment, each residue of the sense strand and the antisense strand is independently modified with LNA, HNA, CeNA, 2'-methoxyethyl, 2'-O-methyl, 2'-O-allyl, 2'-C-allyl, 2'-deoxy, 2'-hydroxyl, or 2'-fluoro. The strand may contain two or more modifications. In one embodiment, each residue of the sense strand and the antisense strand is independently modified with 2'-O-methyl or 2'-fluoro.

[0207] Typically, at least two different modifications are present in the sense strand and the antisense strand. Those two modifications can be 2'-O-methyl or 2'-fluoro modifications, or others.

[0208] In one embodiment, N a and / or N b contains an alternating pattern of modifications. As used herein, the term "alternating motif" refers to a motif having one or more modifications, with each modification being present at alternating nucleotides of one strand. Alternating nucleotides can refer to every other nucleotide, every third nucleotide, or a similar pattern. For example, if A, B, and C each represent one type of modification to a nucleotide, the alternating motif can be "ABABABABABAB···", "AABBAABBAABB···", "AABAABAABAAB···", "AAABAAABAAAB···", "AAABBBAAABBB···", or "ABCABCABCABC···", etc.

[0209] The types of modifications included in the alternating motif can be the same or different. For example, if A, B, C, D each represent one type of modification on a nucleotide, the alternating pattern, i.e., the modification at every other nucleotide, may be the same, but each of the sense strand or the antisense strand can be selected from several possibilities of modifications within an alternating motif such as "ABABAB···", "ACACAC···", "BDBDBD···", or "CDCDCD···".

[0210] In one embodiment, the RNAi agent of the present invention includes a modified pattern of alternating motifs in the sense strand that is shifted relative to the modified pattern of alternating motifs in the antisense strand. This shift can be such that the modified groups of the nucleotides of the sense strand correspond to different modified groups of the nucleotides of the antisense strand, or vice versa. For example, when the sense strand is paired with the antisense strand in the dsRNA duplex, the alternating motifs in the sense strand may start with "ABABAB" from the 5' to 3' of the strand, and the alternating motifs in the antisense strand may be started with "BABABA" from the 5' to 3' of the strand within the duplex region. As another example, the alternating motifs in the sense strand may start with "AABBAABB" from the 5' to 3' of the strand, and the alternating motifs in the antisense strand may be started with "BBAABBAA" from the 5' to 3' of the strand within the duplex region, thereby having a complete or partial shift in the modified pattern between the sense strand and the antisense strand.

[0211] In one embodiment, the RNAi agent includes a pattern of alternating motifs of 2'-O-methyl modification and 2'-F modification in the sense strand, and this pattern has a shift relative to the pattern of alternating motifs of 2'-O-methyl modification and 2'-F modification in the antisense strand, that is, the 2'-O-methyl modified nucleotides in the sense strand form base pairs with the 2'-F modified nucleotides in the antisense strand, and vice versa. The 1st position of the sense strand may start with a 2'-F modification, and the 1st position of the antisense strand may start with a 2'-O-methyl modification.

[0212] The introduction of one or more motifs of three identical modifications on three consecutive nucleotides into the sense strand and / or the antisense strand interrupts the initial modified pattern present in the sense strand and / or the antisense strand. This interruption of the modified pattern of the sense strand and / or the antisense strand by introducing one or more motifs of three identical modifications on three consecutive nucleotides unexpectedly enhances the gene silencing activity against the target gene.

[0213] In one embodiment, when a motif of three identical modifications on three consecutive nucleotides is introduced into either strand, the modification of the nucleotide adjacent to the motif is a modification different from the modification of the motif. For example, a part of the sequence containing the motif is "···N a YYYN b ···", where "Y" represents the modification of the motif of three identical modifications in three consecutive nucleotides, and "N a " and "N b " represent the modifications of the nucleotides adjacent to the motif "YYY" that are different from the modification of Y, and N a and N b can be the same or different modifications. Alternatively, N a and / or N b may or may not be present when a wing modification is present.

[0214] The RNAi agent may further include at least one phosphorothioate or methylphosphonate nucleotide internucleotide linkage. The modification of the phosphorothioate or methylphosphonate nucleotide internucleotide linkage may be present in any nucleotide of the sense strand, the antisense strand, or both strands at any position of the strand. For example, the modification of the internucleotide linkage may be present in all nucleotides in the sense strand and / or the antisense strand; the modification of each internucleotide linkage may be present in an alternating pattern in the sense strand and / or the antisense strand; or the sense strand or the antisense strand may include the modification of both internucleotide linkages in an alternating pattern. The alternating pattern of the modification of the internucleotide linkage in the sense strand may be the same as or different from that in the antisense strand, and the alternating pattern of the modification of the internucleotide linkage in the sense strand may have a shift relative to the alternating pattern of the modification of the internucleotide linkage in the antisense strand. In one embodiment, the double-stranded RNAi agent includes 6 to 8 phosphorothioate nucleotide internucleotide linkages. In one embodiment, the antisense strand includes two phosphorothioate nucleotide internucleotide linkages at the 5' end and two phosphorothioate nucleotide internucleotide linkages at the 3' end, and the sense strand includes at least two phosphorothioate nucleotide internucleotide linkages at the 5' end or the 3' end.

[0215] In one embodiment, the RNAi comprises a modification of the phosphorothioate or methylphosphonate internucleotide linkage in the overhang region. For example, the overhang region may include two nucleotides having a phosphorothioate or methylphosphonate internucleotide linkage between the two nucleotides. The modification of the internucleotide linkage may also be formed to bind the overhang nucleotide to the paired nucleotide at the end within the double-stranded region. For example, at least two, three, four, or all of the overhang nucleotides may be linked by phosphorothioate or methylphosphonate internucleotide linkages, and optionally, there may be additional phosphorothioate or methylphosphonate internucleotide linkages that bind the overhang nucleotide to the paired nucleotide adjacent to the overhang nucleotide. For example, there may be at least two phosphorothioate internucleotide linkages between the three terminal nucleotides, two of the three nucleotides being overhang nucleotides and the third nucleotide being a paired nucleotide adjacent to the overhang nucleotide. These three terminal nucleotides may be present at the 3' end of the antisense strand, the 3' end of the sense strand, the 5' end of the antisense strand, and / or the 5' end of the antisense strand.

[0216] In one embodiment, the two-nucleotide overhang is at the 3' end of the antisense strand, there are two phosphorothioate internucleotide linkages between the three terminal nucleotides, two of the three nucleotides being overhang nucleotides and the third nucleotide being a paired nucleotide adjacent to the overhang nucleotide. Optionally, the RNAi agent may further have two phosphorothioate internucleotide linkages between the three terminal nucleotides at both the 5' end of the sense strand and the 5' end of the antisense strand.

[0217] In one embodiment, the RNAi agent comprises a mismatch with the target, a mismatch within the double strand, or a combination thereof. The mismatch can occur in the overhang region or the double strand region. Base pairs can be evaluated based on their tendency to promote dissociation or melting (e.g., with respect to the free energy of binding or dissociation of a particular pairing, and the simplest approach is to examine the pairs for each individual pair, although similar or analogous analyses can also be used). With regard to promoting dissociation: A:U is preferred over G:C; G:U is preferred over G:C; I:C is preferred over G:C (I = inosine). Mismatches, e.g., non-canonical or non-standard pairings (as described elsewhere herein), are preferred over canonical (A:T, A:U, G:C) pairings; pairings containing universal bases are preferred over canonical pairings.

[0218] In one embodiment, the RNAi agent comprises at least one of the first 1, 2, 3, 4, or 5 base pairs within the double strand region from the 5' end of the antisense strand independently selected from the group of A:U, G:U, I:C, and a mismatch pair for promoting dissociation of the antisense strand at the 5' end of the double strand, e.g., a non-canonical or non-standard pairing or a pairing containing a universal base.

[0219] In one embodiment, the nucleotide at position 1 within the double strand region from the 5' end of the antisense strand is selected from the group consisting of A, dA, dU, U, and dT. Alternatively, at least one of the first 1, 2, or 3 base pairs within the double strand region from the 5' end of the antisense strand is an AU base pair. For example, the first base pair within the double strand region from the 5' end of the antisense strand is an AU base pair.

[0220] In another embodiment, the nucleotide at the 3' end of the sense strand is deoxy-thymine (dT). In another embodiment, the nucleotide at the 3' end of the antisense strand is deoxy-thymine (dT). In one embodiment, there is a short sequence of deoxy-thymine nucleotides, e.g., two dT nucleotides at the 3' end of the sense strand and / or the antisense strand.

[0221] In one embodiment, the sense strand sequence has the formula (I): 5’n p -N a -(XXX) i -N b -YYY-N b -(ZZZ) j -N a -n q 3’(I) (wherein: i and j are each independently 0 or 1; p and q are each independently 0 to 6; each N a represents an oligonucleotide sequence containing 0 to 25 modified nucleotides, and each sequence contains at least two different modified nucleotides; each N b represents an oligonucleotide sequence containing 0 to 10 modified nucleotides; each n p and n q each represent an overhang nucleotide; where Nb and Y do not have the same modification; XXX, YYY and ZZZ each independently represent one motif of three identical modifications in three consecutive nucleotides) and can be represented by. Preferably, YYY are all 2'-F modified nucleotides.

[0222] In one embodiment, N a and / or N b contains modifications in an alternating pattern.

[0223] In one embodiment, the YYY motif is present at or near the cleavage site of the sense strand. For example, when the RNAi agent has a double-stranded region 17 to 23 nucleotides in length, the YYY motif may be present at or near the cleavage site of the sense strand starting from the first nucleotide from the 5'-end; or optionally, starting from the first paired nucleotide within the double-stranded region from the 5'-end (e.g., it may be present at the 6th, 7th, 8th, 7th, 8th, 9th, 8th, 9th, 10th, 9th, 10th, 11th, 10th, 11th, 12th or 11th, 12th, 13th positions).

[0224] In one embodiment, i is 1 and j is 0, or i is 0 and j is 1, or both i and j are 1. Thus, the sense strand can be represented by the following formula: 5’n p -N a -YYY-N b -ZZZ-N a -n q 3’(Ib); 5’n p -N a -XXX-N b -YYY-N a -n q 3’(Ic); or 5’n p -N a -XXX-N b -YYY-N b -ZZZ-N a -n q 3’(Id).

[0225] When the sense strand is represented by formula (Ib), N b represents an oligonucleotide sequence containing 0 to 10, 0 to 7, 0 to 5, 0 to 4, 0 to 2 or 0 modified nucleotides. Each N a can independently represent an oligonucleotide sequence containing 2 to 20, 2 to 15, or 2 to 10 modified nucleotides.

[0226] When the sense strand is represented as formula (Ic), N brepresents an oligonucleotide sequence containing modified nucleotides of 0 to 10, 0 to 7, 0 to 10, 0 to 7, 0 to 5, 0 to 4, 0 to 2, or 0. Each N a independently may represent an oligonucleotide sequence containing modified nucleotides of 2 to 20, 2 to 15, or 2 to 10.

[0227] When the sense strand is represented by formula (Id), each N b independently represents an oligonucleotide sequence containing modified nucleotides of 0 to 10, 0 to 7, 0 to 5, 0 to 4, 0 to 2, or 0. Preferably, N b is 0, 1, 2, 3, 4, 5, or 6. Each N a independently may represent an oligonucleotide sequence containing modified nucleotides of 2 to 20, 2 to 15, or 2 to 10.

[0228] Each of X, Y, and Z may be the same as or different from each other.

[0229] In other embodiments, i is 0, j is 0, and the sense strand may be represented by the following formula: 5’n p -N a -YYY-N a -n q 3’(Ia).

[0230] When the sense strand is represented by formula (Ia), each N a independently may represent an oligonucleotide sequence containing modified nucleotides of 2 to 20, 2 to 15, or 2 to 10.

[0231] In one embodiment, the antisense strand sequence of RNAi is formula (II): 5’n q’ -N a ’-(Z’Z’Z’) k -N b ’-Y’Y’Y’-N b ’-(X’X’X’) l -N’ a -n p ’3’(II) (wherein: k and l are each independently 0 or 1; p’ and q’ are each independently from 0 to 6; each N a ’ represents an oligonucleotide sequence containing from 0 to 25 modified nucleotides, each sequence containing at least two different modified nucleotides; each N b ’ represents an oligonucleotide sequence containing from 0 to 10 modified nucleotides; each n p ’ and n q ’ each independently represent overhang nucleotides; wherein, N b ’ and Y’ do not have the same modification; X’X’X’, Y’Y’Y’ and Z’Z’Z’ each independently represent one motif of three identical modifications in three consecutive nucleotides) can be represented by.

[0232] In one embodiment, N a ’ and / or N b ’ contain an alternating pattern of modifications.

[0233] The Y’Y’Y’ motif is present at or near the cleavage site of the antisense strand. For example, when the RNAi agent has a double-stranded region 17 to 23 nucleotides in length, the Y’Y’Y’ motif starts counting from the first nucleotide from the 5’ end; or optionally, starting from the 5’ end, counting from the first paired nucleotide within the double-stranded region, it can be present at positions 9, 10, 11; 10, 11, 12; 11, 12, 13; 12, 13, 14; or 13, 14, 15 of the antisense strand. Preferably, the Y’Y’Y’ motif is present at positions 11, 12, 13.

[0234] In one embodiment, the Y’Y’Y’ motif consists entirely of 2’-OMe modified nucleotides.

[0235] In one embodiment, k is 1, l is 0, or k is 0, l is 1, or both k and l are 1.

[0236] Therefore, the antisense strand can be represented by the following formula: 5’n q’ -N a ’-Z’Z’Z’-N b ’-Y’Y’Y’-N a ’-n p’ 3’(IIb); 5’n q’ -N a ’-Y’Y’Y’-N b ’-X’X’X’-n p’ 3’(IIc); or 5’n q’ -N a ’-Z’Z’Z’-N b ’-Y’Y’Y’-N b ’-X’X’X’-N a ’-n p’ 3’(IId).

[0237] When the antisense strand is represented by formula (IIb), N b ’ represents an oligonucleotide sequence containing modified nucleotides of 0 to 10, 0 to 7, 0 to 10, 0 to 7, 0 to 5, 0 to 4, 0 to 2, or 0. Each N a ’ independently represents an oligonucleotide sequence containing modified nucleotides of 2 to 20, 2 to 15, or 2 to 10.

[0238] When the antisense strand is represented by formula (IIc), N b ’ represents an oligonucleotide sequence containing modified nucleotides of 0 to 10, 0 to 7, 0 to 10, 0 to 7, 0 to 5, 0 to 4, 0 to 2, or 0. Each N a ’ independently represents an oligonucleotide sequence containing modified nucleotides of 2 to 20, 2 to 15, or 2 to 10.

[0239] When the antisense strand is represented by formula (IId), each N b' independently represents an oligonucleotide sequence containing 0 to 10, 0 to 7, 0 to 10, 0 to 7, 0 to 5, 0 to 4, 0 to 2, or 0 modified nucleotides. Each N a ' independently represents an oligonucleotide sequence containing 2 to 20, 2 to 15, or 2 to 10 modified nucleotides. Preferably, N b is 0, 1, 2, 3, 4, 5, or 6.

[0240] In other embodiments, k is 0, l is 0, and the antisense strand can be represented by the following formula: 5’n p’ -N a’ -Y’Y’Y’-N a’ -n q’ 3’(Ia).

[0241] When the antisense strand is represented as formula (IIa), each N a ' independently represents an oligonucleotide sequence containing 2 to 20, 2 to 15, or 2 to 10 modified nucleotides.

[0242] Each of X’, Y’, and Z’ can be the same as or different from each other.

[0243] Each nucleotide of the sense strand and the antisense strand can be independently modified with LNA, HNA, CeNA, 2'-methoxyethyl, 2'-O-methyl, 2'-O-allyl, 2'-C-allyl, 2'-hydroxyl, or 2'-fluoro. For example, each nucleotide of the sense strand and the antisense strand is independently modified with 2'-O-methyl or 2'-fluoro. Each X, Y, Z, X’, Y’, and Z’ can in particular represent a 2'-O-methyl modification or a 2'-fluoro modification.

[0244] In one embodiment, for the sense strand of the RNAi agent, when the double-stranded region is 21 nucleotides, starting from the first nucleotide from the 5'-end; or optionally, starting from the 5'-end and counting from the first paired nucleotide within the double-stranded region, it may contain the YYY motif present at positions 9, 10, and 11 of the strand; Y represents a 2'-F modification. The sense strand may further contain an XXX motif or a ZZZ motif as a wing modification at the opposite end of the double-stranded region; XXX and ZZZ each independently represent a 2'-OMe modification or a 2'-F modification.

[0245] In one embodiment, for the antisense strand, starting from the first nucleotide from the 5'-end; or optionally, starting from the 5'-end and counting from the first paired nucleotide within the double-stranded region, it may contain the Y'Y'Y' motif present at positions 11, 12, and 13 of the strand; Y' represents a 2'-O-methyl modification. The antisense strand may further contain an X'X'X' motif or a Z'Z'Z' motif as a wing modification at the opposite end of the double-stranded region; X'X'X' and Z'Z'Z' each independently represent a 2'-OMe modification or a 2'-F modification.

[0246] The sense strand represented by any one of the above formulas (Ia), (Ib), (Ic), and (Id) forms a double strand with the antisense strand represented by any one of the formulas (IIa), (IIb), (IIc), and (IId), respectively.

[0247] Therefore, the RNAi agent for use in the method of the present invention may contain a sense strand and an antisense strand, each strand having 14 to 30 nucleotides, and the RNAi double strand has the formula (III): Sense: 5’n p -N a -(XXX) i -N b -YYY-N b -(ZZZ) j -N a -n q 3’ Antisense: 3’n p’ -N a ’ -(X’X’X’) k -N b ’ -Y’Y’Y’-N b ’ -(Z’Z’Z’) l -N a ’ -n q ’ 5’ (III) (wherein: i, j, k, and l are each independently 0 or 1; p, p’, q, and q’ are each independently 0 to 6; each N a and N a ’ represents an oligonucleotide sequence containing 0 to 25 modified nucleotides, and each sequence contains at least two different modified nucleotides; each N b and N b ’ represents an oligonucleotide sequence containing 0 to 10 modified nucleotides; here, each n, which may or may not be present, p n’ p n q n’ q and n represents an overhang nucleotide independently; XXX, YYY, ZZZ, X’X’X’, Y’Y’Y’, and Z’Z’Z’ each independently represent one motif of three identical modifications on three consecutive nucleotides) is represented by.

[0248] In one embodiment, i is 0 and j is 0; or i is 1 and j is 0; or i is 0 and j is 1; or both i and j are 0; or both i and j are 1. In another embodiment, k is 0 and l is 0; or k is 1 and l is 0; k is 0 and l is 1; or both k and l are 0; or both k and l are 1.

[0249] Exemplary combinations of the sense and antisense strands forming the RNAi double strand include the following formula: 5’n p -N a -YYY-N a -n q 3’ 3’n p ’ -N a ’ -Y’Y’Y’-N a ’ n q ’ 5’ (IIIa) 5’n p -N a -YYY-N b -ZZZ-N a -n q 3’ 3’n p ’ -N a ’ -Y’Y’Y’-N b ’ -Z’Z’Z’-N a ’ n q ’ 5’ (IIIb) 5’n p -N a -XXX-N b -YYY-N a -n q 3’ 3’n p ’ -N a ’ -X’X’X’-N b ’ -Y’Y’Y’-N a’ -n q ’ 5' (IIIc) 5'n p -N a -XXX-N b -YYY-N b -ZZZ-N a -n q 3' 3'n p ’ -N a ’ -X’X’X’-N b ’ -Y’Y’Y’-N b ’ -Z’Z’Z’-N a -n q ’ 5' (IIId)

[0250] When the RNAi agent is represented by formula (IIIa), each N a independently represents an oligonucleotide sequence containing 2 to 20, 2 to 15, or 2 to 10 modified nucleotides.

[0251] When the RNAi agent is represented by formula (IIIb), each N b independently represents an oligonucleotide sequence containing 1 to 10, 1 to 7, 1 to 5, or 1 to 4 modified nucleotides. Each N a independently represents an oligonucleotide sequence containing 2 to 20, 2 to 15, or 2 to 10 modified nucleotides.

[0252] When the RNAi agent is represented by formula (IIIc), each N b and N b ' independently represent an oligonucleotide sequence containing 0 to 10, 0 to 7, 0 to 10, 0 to 7, 0 to 5, 0 to 4, 0 to 2, or 0 modified nucleotides. Each N a independently represents an oligonucleotide sequence containing 2 to 20, 2 to 15, or 2 to 10 modified nucleotides.

[0253] When the RNAi agent is represented by formula (IIId), each N b , N b ’ independently represents an oligonucleotide sequence containing 0 to 10, 0 to 7, 0 to 10, 0 to 7, 0 to 5, 0 to 4, 0 to 2, or 0 modified nucleotides. Each N a , N a ’ independently represents an oligonucleotide sequence containing 2 to 20, 2 to 15, or 2 to 10 modified nucleotides. N a , N a ’, N b and N b ’ each independently contains modifications in an alternating pattern.

[0254] Each of X, Y, and Z in formulas (III), (IIIa), (IIIb), (IIIc), and (IIId) can be the same as or different from each other.

[0255] When the RNAi agent is represented by formulas (III), (IIIa), (IIIb), (IIIc), and (IIId), at least one of the Y nucleotides can form a base pair with one of the Y’ nucleotides. Alternatively, at least two of the Y nucleotides can form base pairs with the corresponding Y’ nucleotides; or all three of the Y nucleotides can all form base pairs with the corresponding Y’ nucleotides.

[0256] When the RNAi agent is represented by formula (IIIb) or (IIId), at least one of the Z nucleotides can form a base pair with one of the Z’ nucleotides. Alternatively, at least two of the Z nucleotides can form base pairs with the corresponding Z’ nucleotides; or all three of the Z nucleotides can all form base pairs with the corresponding Z’ nucleotides.

[0257] When the RNAi agent is represented by formula (IIIc) or (IIId), at least one of the X nucleotides can form a base pair with one of the X' nucleotides. Alternatively, at least two of the X nucleotides form base pairs with the corresponding X' nucleotides; or all three of the X nucleotides all form base pairs with the corresponding X' nucleotides.

[0258] In one embodiment, the modification on the Y nucleotide is different from the modification on the Y' nucleotide, the modification on the Z nucleotide is different from the modification on the Z' nucleotide, and / or the modification on the X nucleotide is different from the modification on the X' nucleotide.

[0259] In one embodiment, when the RNAi agent is represented by formula (IIId), N a The modification is a 2'-O-methyl or 2'-fluoro modification. In another embodiment, when the RNAi agent is represented by formula (IIId), N a The modification is a 2'-O-methyl or 2'-fluoro modification, and n p '>0, and at least one n p ' is linked to the adjacent nucleotide via a phosphorothioate bond. In yet another embodiment, when the RNAi agent is represented by formula (IIId), N a The modification is a 2'-O-methyl or 2'-fluoro modification, and n p '>0, and at least one n p ' is linked to the adjacent nucleotide via a phosphorothioate bond, and the sense strand is conjugated to one or more GalNAc derivatives linked via a divalent or trivalent branched linker (described below). In another embodiment, when the RNAi agent is represented by formula (IIId), N a The modification is a 2'-O-methyl or 2'-fluoro modification, and n p '>0, and at least one n p' is linked to an adjacent nucleotide via a phosphorothioate bond, the sense strand contains at least one phosphorothioate bond, and the sense strand is conjugated to one or more GalNAc derivatives linked via a divalent or trivalent branched linker.

[0260] In one embodiment, when the RNAi agent is represented by formula (IIIa), N a The modification is a 2'-O-methyl or 2'-fluoro modification, and n p '> 0, and at least one n p ' is linked to an adjacent nucleotide via a phosphorothioate bond, the sense strand contains at least one phosphorothioate bond, and the sense strand is conjugated to one or more GalNAc derivatives linked via a divalent or trivalent branched linker.

[0261] In one embodiment, the RNAi agent is a multimer comprising at least two double strands represented by formula (III), (IIIa), (IIIb), (IIIc), and (IIId), and the double strands are linked by a linker. The linker can be cleavable or non-cleavable. Optionally, the multimer further comprises a ligand. Each of the double strands can target the same gene or two different genes; or each of the double strands can target the same gene at two different target sites.

[0262] In one embodiment, the RNAi agent is a multimer comprising 3, 4, 5, 6 or more double strands represented by formula (III), (IIIa), (IIIb), (IIIc), and (IIId), and the double strands are linked by a linker. The linker can be cleavable or non-cleavable. Optionally, the multimer further comprises a ligand. Each of the double strands can target the same gene or two different genes; or each of the double strands can target the same gene at two different target sites.

[0263] In one embodiment, two RNAi agents represented by formulas (III), (IIIa), (IIIb), (IIIc), and (IIId) are bound to each other at one or both of the 5'-end and the 3'-end and are optionally conjugated to a ligand. Each of the RNAi agents can target the same gene or two different genes; or each of the RNAi agents can target the same gene at two different target sites.

[0264] A variety of publications describe multimeric RNAi agents that can be used in the methods of the present invention. Such publications include WO 2007 / 091269 pamphlet, US Patent No. 7858769 specification, WO 2010 / 141511 pamphlet, WO 2007 / 117686 pamphlet, WO 2009 / 014887 pamphlet, and WO 2011 / 031520 pamphlet, the entire contents of each of which are incorporated herein by reference.

[0265] As described in more detail below, RNAi agents comprising conjugation of one or more carbohydrate moieties to the RNAi agent can optimize one or more properties of the RNAi agent. In many cases, the carbohydrate moiety is attached to a modified subunit of the RNAi agent. For example, the ribose sugar of one or more ribonucleotide subunits of a dsRNA agent can be replaced with another moiety, for example, a non-carbohydrate (preferably cyclic) carrier to which a carbohydrate ligand is attached. A ribonucleotide subunit in which the ribose sugar of the subunit is thus replaced is referred to herein as a ribose substitution modified subunit (RRMS). The cyclic carrier may be a carbocyclic system, i.e., all ring atoms are carbon atoms, or a heterocyclic system, i.e., one or more ring atoms may be heteroatoms, such as nitrogen, oxygen, sulfur. The cyclic carrier may be a monocyclic system or may contain two or more rings, such as fused rings. The cyclic carrier may be a completely saturated ring system or may contain one or more double bonds.

[0266] The ligand can be bound to the polynucleotide via a carrier. The carrier includes (i) at least one "backbone attachment point", preferably two "backbone attachment points" and (ii) at least one "tethering attachment point". As used herein, a "backbone attachment point" refers to a functional group, such as a hydroxyl group, or generally a backbone, such as a phosphate, or a modified phosphate, such as a sulfur-containing phosphate, that is available for and suitable for incorporation of the carrier into the backbone of a ribonucleic acid and forms a suitable bond therewith. A "tethering attachment point" (TAP) refers, in certain embodiments, to a ring-forming atom of a cyclic carrier that connects a selected moiety, such as a carbon atom or a heteroatom (different from the atom providing the backbone attachment point). This moiety can be, for example, a carbohydrate, such as a monosaccharide, disaccharide, trisaccharide, tetrasaccharide, oligosaccharide, and polysaccharide. Optionally, the selected moiety is connected to the cyclic carrier by an intervening tether. Thus, the cyclic carrier often contains a functional group, such as an amino group, or generally provides a bond suitable for incorporation or tethering of another chemical component, such as a ligand, into the ring structure.

[0267] The RNAi agent may be conjugated to the ligand via a carrier, which may be a cyclic group or a cyclic group; preferably, the cyclic group is selected from pyrrolidinyl, pyrazolinyl, pyrazolidinyl, imidazolinyl, imidazolidinyl, piperidinyl, piperazinyl, [1,3]dioxolane, oxazolidinyl, isoxazolidinyl, morpholinyl, thiazolidinyl, isothiazolidinyl, quinoxalinyl, pyridazinonyl, tetrahydrofuryl, and decalin; preferably, the cyclic group is selected from a serinol backbone or a diethanolamine backbone.

[0268] In certain embodiments, the RNAi agent for use in the methods of the invention is an agent selected from the group of agents listed in any one of Tables 3, 4, 5, 6, 18, 19, 20, 21, and 23. These agents may further include a ligand.

[0269] IV. iRNA Conjugated to a Ligand Another modification of the RNA of the iRNA of the present invention involves chemically binding to the RNA one or more ligands, moieties or conjugates that improve the activity, cellular distribution or cellular uptake of the iRNA.Such moieties include, but are not limited to, cholesterol moieties (Letsinger et al., Proc. Natl. Acid. Sci. USA, 1989, 86: 6553-6556), cholic acid (Manoharan et al., Biorg. Med. Chem. Let., 1994, 4: 1053-1060), thioethers such as beryl-S-tritylthiol (Manoharan et al., Ann. N.Y. Acad. Sci., 1992, 660: 306-309; Manoharan et al., Biorg. Med. Chem. Let., 1993, 3: 2765-2770), thiocolesterol (Oberhauser et al., Nucl. Acids Res., 1992, 20: 533-538), aliphatic chains such as dodecanediol or undecyl residues (Saison-Behmoaras et al., EMBO J, 1991, 10: 1111-1118; Kabanov et al., FEBS Lett., 1990, 259: 327-330; Svinarchuk et al., Biochimie, 1993, 75: 49-54), phospholipids such as di-hexadecyl-rac-glycerol or triethyl-a...

Claims

1. 1. A method for treating a subject suffering from paroxysmal nocturnal hemoglobinuria (PNH), comprising: administering to an eculizumab naive subject a fixed dose of 200-400 mg once weekly of a double-stranded ribonucleic acid (dsRNA) agent for inhibiting expression of complement component C5; administering to the subject a dose of about 300 mg to about 900 mg of eculizumab, or an antigen-binding fragment thereof, thereby treating the subject; wherein the dsRNA agent comprises a sense strand and an antisense strand; wherein the sense strand comprises 5'-asasGfcAfaGfaUfAfUfuUfuuAfuAfaua-3' (SEQ ID NO: 2876) and the antisense strand comprises 5'-usAfsUfuAfuaAfaAfauaUfcUfuGfcuususudTdT-3' (SEQ ID NO: 2889); wherein a, g, c and u are 2'-O-methyl (2'-OMe) A, G, C and U, respectively; Af, Gf, Cf and Uf are 2'-fluoro A, G, C and U, respectively; dT is a deoxy-thymine nucleotide; and s is a phosphorothioate linkage.

2. 1. A method for treating a subject suffering from paroxysmal nocturnal hemoglobinuria (PNH), comprising: administering to an eculizumab naive subject a fixed dose of 200-400 mg once per month of a double-stranded ribonucleic acid (dsRNA) agent for inhibiting expression of complement component C5; administering to the subject a dose of about 300 mg to about 900 mg of eculizumab, or an antigen-binding fragment thereof, thereby treating the subject; wherein the dsRNA agent comprises a sense strand and an antisense strand; wherein the sense strand comprises 5'-asasGfcAfaGfaUfAfUfuUfuuAfuAfaua-3' (SEQ ID NO: 2876) and the antisense strand comprises 5'-usAfsUfuAfuaAfaAfauaUfcUfuGfcuususudTdT-3' (SEQ ID NO: 2889); wherein a, g, c and u are 2'-O-methyl (2'-OMe) A, G, C and U, respectively; Af, Gf, Cf and Uf are 2'-fluoro A, G, C and U, respectively; dT is a deoxy-thymine nucleotide; and s is a phosphorothioate linkage.

3. 1. A method for treating a subject suffering from paroxysmal nocturnal hemoglobinuria (PNH), comprising: administering to an eculizumab naive subject a fixed dose of 200 mg once weekly for 10-15 weeks, followed by a fixed dose of 400 mg once weekly of said dsRNA agent to inhibit expression of complement component C5; administering to the subject a dose of about 300 mg to about 900 mg of eculizumab, or an antigen-binding fragment thereof, thereby treating the subject; wherein the dsRNA agent comprises a sense strand and an antisense strand; wherein the sense strand comprises 5'-asasGfcAfaGfaUfAfUfuUfuuAfuAfaua-3' (SEQ ID NO: 2876) and the antisense strand comprises 5'-usAfsUfuAfuaAfaAfauaUfcUfuGfcuususudTdT-3' (SEQ ID NO: 2889); wherein a, g, c and u are 2'-O-methyl (2'-OMe) A, G, C and U, respectively; Af, Gf, Cf and Uf are 2'-fluoro A, G, C and U, respectively; dT is a deoxy-thymine nucleotide; and s is a phosphorothioate linkage.

4. 1. A method for treating a subject suffering from paroxysmal nocturnal hemoglobinuria (PNH), comprising: administering to an eculizumab naive subject a fixed dose of 200 mg once per month for 2-4 months of a double-stranded ribonucleic acid (dsRNA) agent for inhibiting expression of complement component C5; administering to the subject a dose of about 300 mg to about 900 mg of eculizumab, or an antigen-binding fragment thereof, thereby treating the subject; wherein the dsRNA agent comprises a sense strand and an antisense strand; wherein the sense strand comprises 5'-asasGfcAfaGfaUfAfUfuUfuuAfuAfaua-3' (SEQ ID NO: 2876) and the antisense strand comprises 5'-usAfsUfuAfuaAfaAfauaUfcUfuGfcuususudTdT-3' (SEQ ID NO: 2889); wherein a, g, c and u are 2'-O-methyl (2'-OMe) A, G, C and U, respectively; Af, Gf, Cf and Uf are 2'-fluoro A, G, C and U, respectively; dT is a deoxy-thymine nucleotide; and s is a phosphorothioate linkage.

5. 1. A method for treating a subject suffering from paroxysmal nocturnal hemoglobinuria (PNH), comprising: administering to an eculizumab naive subject a fixed dose of 400 mg once weekly of a double-stranded ribonucleic acid (dsRNA) agent for inhibiting expression of complement component C5; administering to the subject a dose of about 300 mg to about 900 mg of eculizumab, or an antigen-binding fragment thereof, thereby treating the subject; wherein the dsRNA agent comprises a sense strand and an antisense strand; wherein the sense strand comprises 5'-asasGfcAfaGfaUfAfUfuUfuuAfuAfaua-3' (SEQ ID NO: 2876) and the antisense strand comprises 5'-usAfsUfuAfuaAfaAfauaUfcUfuGfcuususudTdT-3' (SEQ ID NO: 2889); wherein a, g, c and u are 2'-O-methyl (2'-OMe) A, G, C and U, respectively; Af, Gf, Cf and Uf are 2'-fluoro A, G, C and U, respectively; dT is a deoxy-thymine nucleotide; and s is a phosphorothioate linkage.

6. 1. A method for treating a subject suffering from paroxysmal nocturnal hemoglobinuria (PNH), comprising: administering to an eculizumab naive subject a fixed dose of 400 mg once monthly of a double-stranded ribonucleic acid (dsRNA) agent for inhibiting expression of complement component C5; administering to the subject a dose of about 300 mg to about 900 mg of eculizumab, or an antigen-binding fragment thereof, thereby treating the subject; wherein the dsRNA agent comprises a sense strand and an antisense strand; wherein the sense strand comprises 5'-asasGfcAfaGfaUfAfUfuUfuuAfuAfaua-3' (SEQ ID NO: 2876) and the antisense strand comprises 5'-usAfsUfuAfuaAfaAfauaUfcUfuGfcuususudTdT-3' (SEQ ID NO: 2889); wherein a, g, c and u are 2'-O-methyl (2'-OMe) A, G, C and U, respectively; Af, Gf, Cf and Uf are 2'-fluoro A, G, C and U, respectively; dT is a deoxy-thymine nucleotide; and s is a phosphorothioate linkage.

7. 1. A method for treating a subject suffering from paroxysmal nocturnal hemoglobinuria (PNH), comprising: administering a fixed dose of 400 mg once weekly of a double-stranded ribonucleic acid (dsRNA) agent to inhibit expression of complement component C5 to a subject previously treated with eculizumab; administering to the subject a dose of about 300 mg to about 900 mg of eculizumab, or an antigen-binding fragment thereof, thereby treating the subject; wherein the dsRNA agent comprises a sense strand and an antisense strand; wherein the sense strand comprises 5'-asasGfcAfaGfaUfAfUfuUfuuAfuAfaua-3' (SEQ ID NO: 2876) and the antisense strand comprises 5'-usAfsUfuAfuaAfaAfauaUfcUfuGfcuususudTdT-3' (SEQ ID NO: 2889); wherein a, g, c and u are 2'-O-methyl (2'-OMe) A, G, C and U, respectively; Af, Gf, Cf and Uf are 2'-fluoro A, G, C and U, respectively; dT is a deoxy-thymine nucleotide; and s is a phosphorothioate linkage.

8. 1. A method for treating a subject suffering from paroxysmal nocturnal hemoglobinuria (PNH), comprising: administering a fixed dose of 400 mg once monthly of a double-stranded ribonucleic acid (dsRNA) agent for inhibiting expression of complement component C5 to a subject previously treated with eculizumab; administering to the subject a dose of about 300 mg to about 900 mg of eculizumab, or an antigen-binding fragment thereof, thereby treating the subject; wherein the dsRNA agent comprises a sense strand and an antisense strand; wherein the sense strand comprises 5'-asasGfcAfaGfaUfAfUfuUfuuAfuAfaua-3' (SEQ ID NO: 2876) and the antisense strand comprises 5'-usAfsUfuAfuaAfaAfauaUfcUfuGfcuususudTdT-3' (SEQ ID NO: 2889); wherein a, g, c and u are 2'-O-methyl (2'-OMe) A, G, C and U, respectively; Af, Gf, Cf and Uf are 2'-fluoro A, G, C and U, respectively; dT is a deoxy-thymine nucleotide; and s is a phosphorothioate linkage.

9. 1. A method for treating a subject suffering from paroxysmal nocturnal hemoglobinuria (PNH), comprising: administering to a subject previously treated with eculizumab a fixed dose of 400 mg once weekly for 2-8 weeks of a double-stranded ribonucleic acid (dsRNA) agent for inhibiting expression of complement component C5; administering to the subject a dose of about 300 mg to about 900 mg of eculizumab, or an antigen-binding fragment thereof, thereby treating the subject; wherein the dsRNA agent comprises a sense strand and an antisense strand; wherein the sense strand comprises 5'-asasGfcAfaGfaUfAfUfuUfuuAfuAfaua-3' (SEQ ID NO: 2876) and the antisense strand comprises 5'-usAfsUfuAfuaAfaAfauaUfcUfuGfcuususudTdT-3' (SEQ ID NO: 2889); wherein a, g, c and u are 2'-O-methyl (2'-OMe) A, G, C and U, respectively; Af, Gf, Cf and Uf are 2'-fluoro A, G, C and U, respectively; dT is a deoxy-thymine nucleotide; and s is a phosphorothioate linkage.

10. 1. A method for treating a subject suffering from paroxysmal nocturnal hemoglobinuria (PNH), comprising: administering a fixed dose of 400 mg once per month for 1-2 months to a subject previously treated with eculizumab of a double-stranded ribonucleic acid (dsRNA) agent for inhibiting expression of complement component C5; administering to the subject a dose of about 300 mg to about 900 mg of eculizumab, or an antigen-binding fragment thereof, thereby treating the subject; wherein the dsRNA agent comprises a sense strand and an antisense strand; wherein the sense strand comprises 5'-asasGfcAfaGfaUfAfUfuUfuuAfuAfaua-3' (SEQ ID NO: 2876) and the antisense strand comprises 5'-usAfsUfuAfuaAfaAfauaUfcUfuGfcuususudTdT-3' (SEQ ID NO: 2889); wherein a, g, c and u are 2'-O-methyl (2'-OMe) A, G, C and U, respectively; Af, Gf, Cf and Uf are 2'-fluoro A, G, C and U, respectively; dT is a deoxy-thymine nucleotide; and s is a phosphorothioate linkage.

11. 1. A method for treating a subject suffering from paroxysmal nocturnal hemoglobinuria (PNH), comprising: administering a fixed dose of 200 mg once weekly of a double-stranded ribonucleic acid (dsRNA) agent for inhibiting expression of complement component C5 to a subject previously treated with eculizumab; administering to the subject a dose of about 300 mg to about 900 mg of eculizumab, or an antigen-binding fragment thereof, thereby treating the subject; wherein the dsRNA agent comprises a sense strand and an antisense strand; wherein the sense strand comprises 5'-asasGfcAfaGfaUfAfUfuUfuuAfuAfaua-3' (SEQ ID NO: 2876) and the antisense strand comprises 5'-usAfsUfuAfuaAfaAfauaUfcUfuGfcuususudTdT-3' (SEQ ID NO: 2889); wherein a, g, c and u are 2'-O-methyl (2'-OMe) A, G, C and U, respectively; Af, Gf, Cf and Uf are 2'-fluoro A, G, C and U, respectively; dT is a deoxy-thymine nucleotide; and s is a phosphorothioate linkage.

12. 1. A method for treating a subject suffering from paroxysmal nocturnal hemoglobinuria (PNH), comprising: administering a fixed dose of 200 mg once monthly of a double-stranded ribonucleic acid (dsRNA) agent for inhibiting expression of complement component C5 to a subject previously treated with eculizumab; administering to the subject a dose of about 300 mg to about 900 mg of eculizumab, or an antigen-binding fragment thereof, thereby treating the subject; wherein the dsRNA agent comprises a sense strand and an antisense strand; wherein the sense strand comprises 5'-asasGfcAfaGfaUfAfUfuUfuuAfuAfaua-3' (SEQ ID NO: 2876) and the antisense strand comprises 5'-usAfsUfuAfuaAfaAfauaUfcUfuGfcuususudTdT-3' (SEQ ID NO: 2889); wherein a, g, c and u are 2'-O-methyl (2'-OMe) A, G, C and U, respectively; Af, Gf, Cf and Uf are 2'-fluoro A, G, C and U, respectively; dT is a deoxy-thymine nucleotide; and s is a phosphorothioate linkage.

13. 1. A method for treating a subject suffering from paroxysmal nocturnal hemoglobinuria (PNH), comprising: administering a fixed dose of 200 mg once weekly of a double-stranded ribonucleic acid (dsRNA) agent for inhibiting expression of complement component C5 to subjects who have not responded to treatment with eculizumab; administering to the subject a dose of about 300 mg to about 900 mg of eculizumab, or an antigen-binding fragment thereof, thereby treating the subject; wherein the dsRNA agent comprises a sense strand and an antisense strand; wherein the sense strand comprises 5'-asasGfcAfaGfaUfAfUfuUfuuAfuAfaua-3' (SEQ ID NO: 2876) and the antisense strand comprises 5'-usAfsUfuAfuaAfaAfauaUfcUfuGfcuususudTdT-3' (SEQ ID NO: 2889); wherein a, g, c and u are 2'-O-methyl (2'-OMe) A, G, C and U, respectively; Af, Gf, Cf and Uf are 2'-fluoro A, G, C and U, respectively; dT is a deoxy-thymine nucleotide; and s is a phosphorothioate linkage.

14. 1. A method for treating a subject suffering from paroxysmal nocturnal hemoglobinuria (PNH), comprising: administering a fixed dose of 200 mg once monthly of a double-stranded ribonucleic acid (dsRNA) agent for inhibiting expression of complement component C5 to subjects who have not responded to treatment with eculizumab; administering to the subject a dose of about 300 mg to about 900 mg of eculizumab, or an antigen-binding fragment thereof, thereby treating the subject; wherein the dsRNA agent comprises a sense strand and an antisense strand; wherein the sense strand comprises 5'-asasGfcAfaGfaUfAfUfuUfuuAfuAfaua-3' (SEQ ID NO: 2876) and the antisense strand comprises 5'-usAfsUfuAfuaAfaAfauaUfcUfuGfcuususudTdT-3' (SEQ ID NO: 2889); wherein a, g, c and u are 2'-O-methyl (2'-OMe) A, G, C and U, respectively; Af, Gf, Cf and Uf are 2'-fluoro A, G, C and U, respectively; dT is a deoxy-thymine nucleotide; and s is a phosphorothioate linkage.

15. 1. A method for treating a subject suffering from paroxysmal nocturnal hemoglobinuria (PNH), comprising: administering a fixed dose of 400 mg once weekly of a double-stranded ribonucleic acid (dsRNA) agent for inhibiting expression of complement component C5 to subjects who have not responded to treatment with eculizumab; administering to the subject a dose of about 300 mg to about 900 mg of eculizumab, or an antigen-binding fragment thereof, thereby treating the subject; wherein the dsRNA agent comprises a sense strand and an antisense strand; wherein the sense strand comprises 5'-asasGfcAfaGfaUfAfUfuUfuuAfuAfaua-3' (SEQ ID NO: 2876) and the antisense strand comprises 5'-usAfsUfuAfuaAfaAfauaUfcUfuGfcuususudTdT-3' (SEQ ID NO: 2889); wherein a, g, c and u are 2'-O-methyl (2'-OMe) A, G, C and U, respectively; Af, Gf, Cf and Uf are 2'-fluoro A, G, C and U, respectively; dT is a deoxy-thymine nucleotide; and s is a phosphorothioate linkage.

16. 1. A method for treating a subject suffering from paroxysmal nocturnal hemoglobinuria (PNH), comprising: administering a fixed dose of 400 mg once monthly of a double-stranded ribonucleic acid (dsRNA) agent for inhibiting expression of complement component C5 to subjects who have not responded to treatment with eculizumab; administering to the subject a dose of about 300 mg to about 900 mg of eculizumab, or an antigen-binding fragment thereof, thereby treating the subject; wherein the dsRNA agent comprises a sense strand and an antisense strand; wherein the sense strand comprises 5'-asasGfcAfaGfaUfAfUfuUfuuAfuAfaua-3' (SEQ ID NO: 2876) and the antisense strand comprises 5'-usAfsUfuAfuaAfaAfauaUfcUfuGfcuususudTdT-3' (SEQ ID NO: 2889); wherein a, g, c and u are 2'-O-methyl (2'-OMe) A, G, C and U, respectively; Af, Gf, Cf and Uf are 2'-fluoro A, G, C and U, respectively; dT is a deoxy-thymine nucleotide; and s is a phosphorothioate linkage.

17. 17. The method of any one of claims 1-16, wherein the dose of eculizumab, or an antigen-binding fragment thereof, is about 25% to about 75% of the eculizumab maintenance level dose.

18. 18. The method of claim 17, wherein the dose of eculizumab, or an antigen-binding fragment thereof, is between 300 mg and 600 mg.

19. 18. The method of claim 17, wherein the dose of eculizumab, or an antigen-binding fragment thereof, is between 600 mg and 900 mg.

20. 17. The method of any one of claims 1-16, wherein the frequency of administration of eculizumab is reduced compared to the frequency of administration required by said level.

21. 21. The method of claim 20, wherein eculizumab is administered to the subject once every 4 weeks, once every 2 months, once every 3 months, once every 4 months, once every 5 months, or once every 6 months.

22. 7. The method of any one of claims 1-6, wherein the eculizumab, or antigen-binding fragment thereof, is administered to the subject as a fixed dose of 600 mg once every four weeks.

23. 7. The method of any one of claims 1-6, wherein the eculizumab, or antigen-binding fragment thereof, is administered to the subject as a fixed dose of 300 mg once every four weeks.

24. 17. The method of any one of claims 7-16, wherein the eculizumab, or antigen-binding fragment thereof, is administered to the subject as a fixed dose of 600 mg once every four weeks.

25. 17. The method of any one of claims 7-16, wherein the eculizumab, or antigen-binding fragment thereof, is administered to the subject as a fixed dose of 900 mg once every four weeks.

26. 13. The method of any one of claims 7-12, wherein the previous treatment with eculizumab, or an antigen-binding fragment thereof, comprised administration to the subject of a fixed dose of 900 mg eculizumab every other week.

27. 17. The method of any one of claims 13-16, wherein the eculizumab, or antigen-binding fragment thereof, treatment to which the subject did not respond comprised administration to the subject of a fixed dose of 1200 mg every other week.

28. 28. The method of any one of claims 1-27, wherein the dsRNA agent and the eculizumab, or antigen-binding fragment thereof, are administered to the subject simultaneously.

29. 28. The method of any one of claims 1-27, wherein the dsRNA agent is administered to the subject prior to the eculizumab, or antigen-binding fragment thereof.

30. 28. The method of any one of claims 1-27, wherein the eculizumab, or antigen-binding fragment thereof, is administered to the subject prior to the dsRNA agent.

31. The method of any one of claims 1 to 27, wherein the treatment prevents a hemolytic attack in the subject.

32. 28. The method of any one of claims 1-27, wherein the treatment reduces mean maximum C5 mRNA levels by at least about 98% relative to baseline.

33. 28. The method of any one of claims 1 to 27, wherein the treatment reduces the minimum residual C5 level to about 1.0 micrograms / ml or less.

34. 28. The method of any one of claims 1-27, wherein the treatment reduces classical complement pathway (CCP) activity by at least about 94% relative to baseline.

35. 28. The method of any one of claims 1-27, wherein the treatment reduces alternative complement pathway (CAP) activity by at least about 94% relative to baseline.

36. 28. The method of any one of claims 1-27, wherein the treatment inhibits mean maximal hemolysis by at least about 75% relative to baseline as measured by inhibition of sheep red blood cell hemolysis.

37. 28. The method of any one of claims 1-27, wherein said treatment reduces the level of lactate dehydrogenase (LDH) in said subject to less than about 1.5 times the upper limit of normal (ULN).

38. The method of any one of claims 7 to 12, wherein the subject previously treated with eculizumab did not have a hemolytic episode.

39. The method of any one of claims 7 to 12, wherein the subject previously treated with eculizumab had a hemolytic episode.

40. The method of any one of claims 1-27, wherein the dsRNA agent is administered to the subject subcutaneously.

41. 28. The method of any one of claims 1-27, wherein the eculizumab is administered intravenously to the subject.

42. 42. The method of any one of claims 1-41, wherein the dsRNA agent further comprises a ligand.

43. 43. The method of claim 42, wherein the ligand is one or more GalNAc derivatives attached via a bivalent or trivalent branched linker.

44. The ligand is 【Chemistry 1】 43. The method of claim 42, wherein:

45. 43. The method of claim 42, wherein the ligand is attached to the 3' end of the sense strand.

46. The RNAi agent is shown in the schematic diagram below. 【Chemistry 2】 46. ​​The method of claim 45, wherein the ligand is conjugated as shown in

47. 1. A method for treating a subject suffering from paroxysmal nocturnal hemoglobinuria (PNH), comprising: administering to an eculizumab naive subject a fixed dose of 200 mg of a double-stranded ribonucleic acid (dsRNA) agent to inhibit expression of complement component C5 once per week for 12 weeks, followed by a fixed dose of 400 mg of said dsRNA agent once per week, thereby treating said subject; wherein the dsRNA agent comprises a sense strand and an antisense strand; wherein the sense strand comprises 5'-asasGfcAfaGfaUfAfUfuUfuuAfuAfaua-3' (SEQ ID NO: 2876) and the antisense strand comprises 5'-usAfsUfuAfuaAfaAfauaUfcUfuGfcuususudTdT-3' (SEQ ID NO: 2889); wherein a, g, c and u are 2'-O-methyl (2'-OMe) A, G, C and U, respectively; Af, Gf, Cf and Uf are 2'-fluoro A, G, C and U, respectively; dT is a deoxy-thymine nucleotide; and s is a phosphorothioate linkage.

48. The method of any one of claims 1 to 47, wherein the treatment is long term.

49. The method of any one of claims 13-16, wherein the dsRNA agent is administered to the subject for 8 to 15 weeks.

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