Compositions and methods for inhibiting the expression of angiotensinogen (AGT) protein

Abstract

This invention provides compositions and methods for inhibiting angiotensinogen (AGT) protein expression for alternative and combination therapies for the treatment of hypertension and other angiotensinogen-related disorders. [Solution] Compositions and methods for inhibiting angiotensinogen (AGT) protein expression are provided. Specifically, compositions and methods are provided that can be used to reduce AGT gene expression and to treat AGT-related diseases and disorders. AGT dsRNA agents, AGT antisense polynucleotide agents, compositions containing AGT dsRNA agents, and compositions containing AGT antisense polynucleotide agents are provided that can be used to reduce AGT expression in cells and subjects.

Description

[Technical Field]

[0001] Some embodiments of the present invention involve the inhibition of angiotensinogen (AGT) protein expression. This relates to compositions and methods useful for [the purpose]. [Background technology]

[0002] The renin-angiotensin-aldosterone system (RAAS) plays a crucial role in blood pressure regulation. The RAAS cascade opens with the secretion of renin into circulation by juxtaglomerular cells of the kidney. It begins. Renin secretion starts in the distal tubules. + Loading, β-sympathetic nerve stimulation and / Alternatively, it can be stimulated by several factors, such as decreased renal blood flow. Active renin in plasma is Angiotensinogen (produced by the liver) is broken down into angiotensin I, and It circulates and, through locally expressed angiotensin-converting enzyme (ACE), It is converted to angiotensin II. The effect of angiotensin II on the RAAS is significant. This portion is exerted by its binding to the angiotensin type II 1 receptor (AT1R), N a + Arterial vasoconstriction, tubular and glomerular effects, such as improved reabsorption or regulation of glomerular filtration rate. It occurs. In addition, other stimuli (adrenocorticotropic hormone, antidiuretic hormone, catecholamines) are present. (including endothelin and serotonin), and Mg 2+ and K + Along with the level, AT1 R stimulation leads to the release of aldosterone, followed by Na in the distal tubules. + and K + Discharge This is promoted.

[0003] For example, this can result in excessive angiotensin II production and / or AT1R stimulation. Dysregulation of the RAAS can cause hypertension and oxidative stress in the heart, kidneys, and arteries, for example. Increased stress promotes inflammation and hypertrophy, for example, left ventricular fibrosis and arterial remodeling. It can also cause ring formation and glomerular sclerosis.

[0004] Hypertension is the most prevalent disease in developed countries, affecting 20-50% of the adult population. It is a disease that is relatively easy to manage. Hypertension is associated with a shortened life expectancy, chronic kidney disease, stroke, and myocardial infarction. Infarction, heart failure, aneurysm (e.g., aortic aneurysm), peripheral vascular disease, cardiac injury (e.g., cardiomegaly or (This includes hypertrophy) and other cardiovascular diseases, disorders and / or medical conditions, etc. It is a major risk factor for various diseases, disorders, and conditions. Furthermore, hypertension is a major risk factor for cardiovascular morbidity. It has been found to be an important risk factor for mortality, accounting for 62% of all strokes, and It accounts for or constitutes 49% of all heart diseases. In 2017, the diagnosis and prevention of hypertension... , and treatment guidelines have been changed to further reduce the risk of hypertension-related diseases and disorders. To that end, even lower blood pressure targets were provided (e.g., Reboussin et al.) .,Systematic Review for the 2017ACC / AHA / AAPA / ABC / ACPM / AGS / APhA / ASH / ASPC / NMA / PCNA Guideline for the Prevention,Detection, Evaluation, and Management of High Blood Pressure in Adults: A Report of the Ameri can College of Cardiology / American Heart Association Task Force on Clinical Prac tice Guidelines.J Am Coll Cardiol.2017 N ov7.pii:S0735-1097(17)41517-8.doi:10.101 6 / j.jacc.2017.11.004; and Whelton et al. (20 17ACC / AHA / AAPA / ABC / ACPM / AGS / APhA / ASH / ASP C / NMA / PCNA Guideline for the Prevention, Detection, Evaluation, and Management of H igh Blood Pressure in Adults:A Report of the American College of Cardiology / Amer ican Heart Association Task Force on Cli nical Practice Guidelines.J Am Coll Card iol.2017 Nov7.pii:S0735-1097(17)41519-1. (see doi:10.1016 / j.jacc.2017.11.006).

[0005] Despite the numerous antihypertensive drugs available to treat high blood pressure, one-third of the target population... In more than two people, symptoms can be controlled with a single type of antihypertensive drug. Therefore, two or more antihypertensive drugs selected from different drug classes are required. Therefore, furthermore, there is a decrease in medication compliance and the side effects experienced with the use of large amounts of drugs. The increase leads to a decrease in the number of people whose blood pressure needs to be controlled.

[0006] Therefore, alternative and adjunctive therapies for the treatment of hypertension and other angiotensinogen-related disorders. Laws are needed in this field of technology. [Overview of the project]

[0007] According to one aspect of the present invention, double-stranded riboblasts inhibit angiotensinogen (AGT) expression. A nucleic acid (dsRNA) drug is provided, and the dsRNA drug comprises a sense strand and an antisense strand It includes, and nucleotide positions 2-18 in the antisense strand are relative to the AGT RNA transcript. and include a complementary region, and that complementary region is one of the antisense sequences shown in Tables 1-4 It contains at least 15 neighboring nucleotides that are different in number 0, 1, 2, or 3. , optionally containing a targeted ligand. In some embodiments, against the AGT RNA transcript The complementary region is one of the antisense sequences shown in Tables 1-4, and differs by 3 nucleotides or less. It contains at least 15, 16, 17, 18, or 19 adjacent nucleotides. In one embodiment, the antisense strand of the dsRNA is one of the target regions of SEQ ID NO: 519 It is at least substantially complementary to the region and is provided in one of Tables 1-4. In one embodiment, the antisense strand of the dsRNA is one of the target regions of SEQ ID NO: 519 It is completely complementary to the domain and is provided in one of Tables 1-4. In some embodiments Furthermore, the dsRNA agent contains one of the sense strand sequences shown in Tables 1-4, and the sense strand sequence The sequence is at least substantially complementary to the antisense strand sequence in the dsRNA agent. In one embodiment, the dsRNA agent comprises one of the sense strand sequences shown in Tables 1-4. Furthermore, its sense strand sequence is perfectly complementary to the antisense strand sequence in dsRNA agents. In some embodiments, the dsRNA agent is one of the sense strand sequences shown in Tables 1-4. Includes. In some embodiments, the dsRNA agent is the sequence shown as a double-strand sequence in Tables 1-4. Includes any of the following.

[0008] In some embodiments, the dsRNA agent is of formula (A): 5'-Z1AGCUUGUUU GUGAAACZ2-3' contains a sense strand with 0, 1, 2, or 3 nucleotides different from the other, In the formula, Z1 is a nucleotide sequence containing 0 to 15 nucleotide motifs, and Z2 is selected from one of A, U, C, or G, or it does not exist. In a particular embodiment... Z1 is a nucleotide sequence containing 1 to 4 nucleotide motifs. In the embodiment, Z1 is a nucleus containing 1, 2, 3, or 4 nucleotide motifs. This is an Otid sequence. In certain embodiments, Z2 is A. In certain embodiments Z1 is a nucleotide sequence containing a CACC or GACC motif. Specific implementation In this state, the Z1 nucleotide sequence has the following motifs: C, AC, UC, GC, CC, ACC, UCC, GCC, CCC, GACC, AACC, UACC, CACC, CGAC C, CCGACC, ACCGACC, AACCGACC, CAACCGACC, CCAA CCGACC, UCCAACCGACC, UUCCAACCGACC, AUUCCAAC Among CGACC, AAUUCCAACCGACC, or GAAUUCCAACCGACC It is selected from one of the following. In some embodiments, the dsRNA agent is formula (B):5'-Z 3GUUUCACAAACAAGCUZ4-3' and 0, 1, 2, or 3 nucleotides The formula includes different antisense chains, where Z3 is selected from one of A, U, C, and G. It is either present or absent, and Z4 contains 0 to 15 nucleotide motifs. This is an ocidal sequence. In certain embodiments, Z4 consists of 1 to 4 nucleotide motifs. It is a nucleotide sequence containing . In certain embodiments, Z4 is 1, 2, 3, or 4 It is a nucleotide sequence containing a nucleotide motif. In certain embodiments, Z 3 is U. In certain embodiments, the Z4 nucleotide sequence is GGUC or GGU Selected from nucleotide sequences containing a G motif. In certain embodiments, the Z4 nucleus The rheotide sequences are based on the following motifs: G, GU, GC, GA, GG, GGU, GGA, GG C, GGG, GGUG, GGUC, GGUU, GGUA, GGUCG, GGUCGG, G GUCGGU, GGUCGGUU, GGUCGGUUG, GGUCGGUUGG, GGU CGGUUGGA, GGUCGGUUGGAA, GGUCGGUUGGAAU, GGUC Select from either GGUUGGAAUU or GGUGGGUUGGAAUUC. In some embodiments, the dsRNA agent comprises a sense strand and an antisense strand, and its The antisense chain and the antisense chain are, respectively, formula (A) and formula (B) as described herein, and 0, 1, 2, or 3 nucleotides contain different nucleotide sequences, and optionally a targeting ligand. Includes. In certain embodiments, the sense strand (A) and antisense strand (B) of the dsRNA agent. Each of these has a length of 35 nucleotides or less. In certain embodiments, Z1 The Z4 nucleotide motif is completely or partially complementary. In some embodiments, In this context, the dsRNA agent is represented by formula (A'): 5'-Z1'CAGCUUGUUUGUGAAA CA-3' contains a sense strand with 0, 1, 2, or 3 nucleotides, and formula (B') :5'-UGUUUCACAAACAAGCUGZ4'-3' and 0, 1, 2 or 3 Nucle Otid contains different antisense chains, where Z1' and Z4' are independently of each other. Contains a 0-13 nucleotide sequence of the creotide motif. In certain embodiments, Z1' And Z4' are, independently, nucleotide motif 1, 2, or 3 nucleotide sequences. Includes. In certain embodiments, the Z1' nucleotide sequence is the following motif: A, U ,G,C,AC,UC,GC,CC,GAC,AAC,UAC,CAC,CGAC,CC GAC, ACCGAC, AACCGAC, CAACCGAC or GAAUUCCAACC It is selected from one of the GACs. The Z4' nucleotide sequence is the following motif: U, C, A, G, GU, GA, GC, GG, GUG, GUC, GUU, GUA, GUCG, G UCGG, GUCGGU, GUCGGUU, GUCGGUUG, or GUCGGUUGGA It will be selected from one of the AUUC options.

[0009] In some embodiments, the dsRNA agent comprises a sense strand and an antisense strand, and anti The sense strand is the antisense sequence shown below. 5'-UACUCAUUAGAAGAAAAGGUG-3'(Sequence ID 162); 5'-UCUUAGACCAAGGAGAAACGG-3'(Sequence ID 163); 5'-UGUUUCACAAACAAGCUGGUC-3'(Sequence ID 167); 5'-UGUUUCACAAACAAGCUGGUG-3'(Sequence ID 523); 5'-UUCGGUUGGAAUUCUUUUUGC-3'(Sequence ID 184); 5'-GUUUCACAAACAAGCUGG-3'(Sequence ID 653); One of them and at least 15 adjacent nucleotides that differ by 0, 1, 2, or 3 nucleotides Contains creotide.

[0010] In some embodiments, the dsRNA agent comprises a sense strand and an antisense strand, and The nucleotide sequences shown below are: Sense chain: 5'-CACCUUUUCUUCUAAUGAGUA-3'(Sequence ID 65) ), Antisense chain: 5'-UACUCAUUAGAAGAAAAGGUG-3' (sequence number) No. 162) One of them and at least 15 adjacent nucleotides that differ by 0, 1, 2, or 3 nucleotides Contains creotide.

[0011] In some embodiments, the dsRNA agent comprises a sense strand and an antisense strand, and The nucleotide sequences shown below are: Sense chain: 5'-CCGUUUCUCCUUGGUCUAAGA-3'(SEQ ID NO: 66) ), Antisense chain: 5'-UCUUAGACCAAGGAGAAACGG-3' (sequence number) No. 163) One of them and at least 15 adjacent nucleotides that differ by 0, 1, 2, or 3 nucleotides Contains creotide.

[0012] In some embodiments, the dsRNA agent comprises a sense strand and an antisense strand, and The nucleotide sequences shown below are: Sense chain: 5'-GACCAGCUUGUUUGUGAAACA-3'(SEQ ID NO: 70) ), Antisense chain: 5'-UGUUUCACAAACAAGCUGGUC-3' (sequence number) No. 167) One of them and at least 15 adjacent nucleotides that differ by 0, 1, 2, or 3 nucleotides Contains creotide.

[0013] In some embodiments, the dsRNA agent comprises a sense strand and an antisense strand, and The nucleotide sequences shown below are: Sense chain: 5'-CACCAGCUUGUUUGUGAAACA-3'(Sequence ID: 5 twenty two) Antisense chain: 5'-UGUUUCACAAACAAGCUGGUG-3'(number: 523); One of them and at least 15 adjacent nucleotides that differ by 0, 1, 2, or 3 nucleotides Contains creotide. In some embodiments, the dsRNA agent comprises a sense strand and an antisense strand, and The nucleotide sequences shown below are: Sense chain: 5'-GCAAAAAGAAUUCCAACCGAA-3'(Sequence ID 87) ), Antisense chain: 5'-UUCGGUUGGAAUUCUUUUUGC-3' (sequence number) No. 184) One of them and at least 15 adjacent nucleotides that differ by 0, 1, 2, or 3 nucleotides Contains creotide.

[0014] In some embodiments, the dsRNA agent comprises a sense strand and an antisense strand, and The nucleotide sequences shown below are: Sense chain: 5'-CCAGCUUGUUUGUGAAAC-3' (SEQ ID NO: 652), Antisense chain: 5'-GUUUCACAAACAAGCUGG-3'(SEQ ID NO: 65) 3) One of them and at least 15 adjacent nucleotides that differ by 0, 1, 2, or 3 nucleotides Contains creotide.

[0015] In some embodiments, the dsRNA double helix is ​​AD00158-19 in Table 1. -2, AD00158-19-1, AD00158-3, AD00158-1, AD00 158-2, AD00158, AD00159, AD00159-1, AD00159- 2, AD00159-19-1, AD00159-19-2, AD00163, AD00 163-1, AD00163-2, AD00163-19-1, AD00163-19- 2, AD00163-3, AD00300-1, AD00300-19-1 and AD00 One of the 300-19-2 double helixes is selected.

[0016] In some embodiments, the dsRNA double helix is ​​AV01227, AV in Table 1. 01228, AV01229, AV01230, AV01231, AV01232, AV 01233, AV01234, AV01235, AV01236, AV01237, AV 01238, AV01239, AV01240, AV01241, AV01242, AV 01243, AV01244, AV01245, AV01246, AV01247, AV 01248, AV01249, AV01250, AV01251, AV01252, AV 01253, AV01254, AV01255, AV01256, AV01257 or A Selected from one of the double hemispheres of V01711.

[0017] In some embodiments, the dsRNA agent comprises at least one modified nucleotide. In certain embodiments, all or substantially all of the nucleotides of the antisense strand are removed. is a modified nucleotide. In some embodiments, at least one of its modified nucleotides Examples of rheotides include 2'-O-methylnucleotide, 2'-fluoronucleotide, and 2' -Deoxynucleotides, 2’,3’-seconucleotide mimics, locke d) Nucleotides, unlocked nucleic acid (UNA) nucleotides, glycol nucleic acid (GNA) Nucleotides, 2’-F-arabinonucleotides, 2’-methoxyethyl nucleotides, Apurinic / apyrimidinic nucleotides, ribitol, inverted nucleotides, inverted apurinic / apyrimidinic nucleotides, inverted 2 ’-OMe nucleotides, inverted 2’-deoxynucleotides, 2’-amino-modified nucleo tides, 2’-alkyl-modified nucleotides, morpholino nucleotides and 3’-OMe nucleo tides, nucleotides containing a 5’-phosphorothioate group, or cholesterol derivatives or terminal nucleotides linked to a dodecyl bisdecyloamide group, 2’-amino-mod ified nucleotides, phosphoramidates or nucleotides containing unnatural bases are included .

[0018] In some embodiments, these double-stranded ribonucleic acid (dsRNA) agents include a sense strand and, an antisense strand that is complementary to at least a portion of the mRNA corresponding to the target gene, and the dsRNA agent includes a nucleotide sequence having an antisense strand represented by formula (C), and a nucleotide sequence having a sense strand represented by formula (D).

[0019] The antisense strand is represented by formula (C) shown in the 3’ to 5’ direction: 3’-(N L ) n N M1 N L N M2 N L N F N L N M3 N L N M4 N L N M5 N M6 NL N M7 N M8 N L N F N L -5' includes formula (C), and The sense chain is shown in equation (D) from 5' to 3': 5'-(N' L ) n 'N' L N' L N' L N' N1 N' N2 N' N3 N' N4 N' F N' L N' N5 N' N6 N' L N' L N' L N' L N' L N' L N' L -3' formula (D) Includes.

[0020] In the above formula, N F Each of these represents a 2'-fluoro-modified nucleotide. M1 , N M 2, N M3 , N M4 , N M5 , N M6 , N M7 and N M8 Independently, modified or unmodified Nu Represents creotide. M1 , N M2 , N M3 , N M4 , N M5 , N M6 , N M7 and N M In 8, there are three 2'-fluoro-modified nucleotides, or only one 2'-fluoro-modified nucleotide. - Modified nucleotides are present. L Each of these is independently a modified or unmodified nucleotide. This represents a modification that is not a 2'-fluoro-modified nucleotide. F Each of them is 2 'Represents a fluoro-modified nucleotide. N' N1 , N' N2 , N' N3 , N' N4 , N ' N5 and N' N6 The 'N' independently represents a modified or unmodified nucleotide. N1 , N' N 2, N' N3 , N' N4 , N' N5 and N' N6 In the 2'-fluoro-modified nucleo There are only two Chidos. N' L Each independently represents a modified or unmodified nucleotide. Furthermore, this modification is not a 2'-fluoro-modified nucleotide. n and n' are independent of each other. Therefore, it can be an integer between 0 and 7.

[0021] In certain embodiments, the anticerene represented by formula (C) in the dsRNA agent Positions 2, 7, 12, 14, and 16 of the chain (from the first pair of nucleotides at the 5' end) The nucleotides of (the nucleotides) are 2'-fluorine-modified nucleotides. The sense strand represented by formula (D) has positions 9, 11 and 13 (the first pair of nu at the 3' end). (Counted from the creotide) is a 2'-fluoro-modified nucleotide. Specific implementation Morphologically, the NM2, NM3, and NM6 positions of the antisense chain represented by formula (C) The nucleotide is a 2'-fluoro-modified nucleotide, represented by formula (D). Sense chain N' N3 , N' N5 The nucleotide at position 2'-fluoro-modified is a nucleotide. be.

[0022] In some embodiments, the dsRNA agent has an E-vinylphosphonate at the 5' end of the guide strand. Contains a nucleotide. In certain embodiments, the dsRNA agent contains at least one It contains phosphorothioate nucleoside interbonding. In certain embodiments, the sense chain is small It contains at least one phosphorothioate nucleoside bond. In some embodiments, The antisense chain contains at least one phosphorothioate nucleoside bond. In one embodiment, the sense chain has 1, 2, 3, 4, 5, or 6 phosphorothioates It contains nucleoside bonds. In some embodiments, the antisense chains are 1, 2, 3, 4 It contains 5 or 6 phosphorothioate nucleoside interbonds.

[0023] In certain embodiments, all or substantially all of the sense chain and antisense chain are removed. All nucleotides are modified nucleotides. In some embodiments, the modified sense strand is The modified sense chain sequences are shown in Tables 2-4. In some embodiments, modified antisense chains are used. The chains are modified antisense chain sequences as shown in Tables 2-4.

[0024] In certain embodiments, the sense chain is complementary or substantially complementary to the antisense chain. The complementary region is 16–23 nucleotides in length. In some embodiments In this embodiment, the complementary region is 19 to 21 nucleotides long. The complementary regions are 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 2 The length is 4, 25, 26, 27, 28, 29, or 30 nucleotides.

[0025] In some embodiments, the length of each chain is 40 nucleotides or less. In this configuration, each strand is 30 nucleotides or less in length. In some embodiments, each strand The length is 25 nucleotides or less. In some embodiments, each strand is 23 nucleotides. The length is less than or equal to 1. In some embodiments, each chain is 4, 15, 16, 17, 18, 1 9, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleos It is the length of Chido.

[0026] In certain embodiments, the dsRNA agent comprises at least one modified nucleotide, It further includes one or more targeted or linking groups. In some embodiments, And one or more targeting groups or linking groups are conjugated to the sense chain. In embodiments, the targeting group or linking group is N-acetyl-galactosamine (GalNAc ) includes. In some embodiments, the targeting group has the following structure: [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] It holds.

[0027] In certain embodiments, the dsRNA agent is conjugated to the 5' end of the sense strand. It contains a targeting group. In some embodiments, the dsRNA agent has a targeting group at the 3' end of the sense strand. It contains a denjugated targeting group. In some embodiments, the antisense chain is 3' It contains an inverted debase residue at the terminal. In certain embodiments, the sense strand is 3' and / or 5'. 'Contains one or two inverted debase residues at the terminal. In some embodiments, dsRNA agent It has two blunt ends. In some embodiments, at least one chain is at least It also includes a 3' overhang that is the length of one nucleotide. In some embodiments, At least one strand contains a 3' overhang that is at least two nucleotides long. .

[0028] In certain embodiments, the present invention relates to open nucleic acids (UNA) used in therapeutic applications. ) Regarding oligomers. Unlocked nucleic acids (UNAs) are the C2' and C3' atoms between ribose molecules. It is an acyclic analog of RNA in which the bonds with atoms are broken. UNA incorporation is tolerable. It has been found that this is beneficial, and in some cases, it can activate siRNA gene silencing activity. Enhance (Meghan A. et al. Locked vs. unlocked n ucleic acids(LNA vs.UNA):contrasting str uctures work towards common therapeutic goals.Chem.Soc.Rev.,2011, 40,5680-5689).

[0029] UNA is a thermally unstable modification, and by replacing a ribonucleotide with UNA... The strength of the base pair and the stability of the double helix are reduced. Seed region of the siRNA antisense strand. The strategic placement of UNAs in [the region] is related to the genetic mediated by microRNAs (miRNAs). It can reduce off-target activity in the sub-silencing mechanism. RNA primarily consists of the antisense seed region (positions 2-8 from the end) and the target mR of gene repression. It recognizes target genes through base pairing with NA. Each miRNA regulates numerous genes. It has the potential to be loaded by the RNA-induced silencing complex (RISC). siRNA antisense strands can also unintentionally cause a large number of transmissions via miRNA-mediated mechanisms. It may be possible to regulate the gene. Therefore, the seed region of the siRNA contains UNA By adding a thermally unstable nucleotide, off-target activity can be reduced. (Lam JK, Chow MY, Zhang Y, Leung SW.si) RNA Versus miRNA as Therapeutics for Gen e Silencing.Mol Ther Nucleic Acids.2015 Sep15;4(9):e252.doi:10.1038 / mtna.2015.23 .PMID:26372022;PMCID:PMC4877448.). In particular, such RNA oligonucleotides or RNA oligonucleotide complexes have a small amount of seed region. It contains at least one UNA nucleotide monomer (Narendra Vaish) et al.Improved specificity of gene silen cing by siRNAs containing unlocked nuclei obase analog.Nucleic Acids Research,2011 (Vol.39, No.5 1823-1832).

[0030] According to the technical solutions of the present invention, RNA oligonucleotides or RNA oligonucleotides Potential advantages of incorporating UNA into the CHIDO complex include, but are not limited to, the following: To be able to 1. Reduction of off-target activity. When UNA is added to the iRNA seed region, the seed region The base pair strength in the region is reduced, thereby causing potential occlusion by micro-RNAs. GET activity decreases. 2. Good tolerance of UNA in terms of siRNA activity. Therefore, UNA may cause an increase in activity.

[0031] Examples of UNA monomers that can be used in technical solutions include, but are not limited to, UNA monomers. but: [ka] These are some examples.

[0032] In some embodiments, the dsRNA agent is the double-stranded AD00158- shown in Tables 2-4. 19-2, AD00158-19-1, AD00158-3, AD00158-1, AD 00158-2, AD00158, AD00159, AD00159-1, AD0015 9-2, AD00159-19-1, AD00159-19-2, AD00163, AD 00163-1, AD00163-2, AD00163-19-1, AD00163-1 9-2, AD00163-3, AD00300-1, AD00300-19-1 and AD It is a modified double helix selected from one of the following: 00300-19-2.

[0033] In some embodiments, the dsRNA agent is the double-stranded AV01227 in Tables 2-4. AV01228, AV01229, AV01230, AV01231, AV01232, AV01233, AV01234, AV01235, AV01236, AV01237, AV01238, AV01239, AV01240, AV01241, AV01242, AV01243, AV01244, AV01245, AV01246, AV01247, AV01248, AV01249, AV01250, AV01251, AV01252, AV01253, AV01254, AV01255, AV01256 and AV01257 It is a modified double helix selected from one of the following:

[0034] According to one aspect of the present invention, any embodiment of the above-described embodiment of the dsRNA agent of the present invention A composition containing is provided. In certain embodiments, the composition is further permitted as a pharmacopoeia. It contains a carrier. In some embodiments, the composition further includes one or more additional The therapeutic agent is included. In certain embodiments, the composition is a kit, container, pack, dispenser. Packaged in a sensor, pre-filled syringe, or vial. In some embodiments... The composition is formulated for subcutaneous or intravenous (IV) administration.

[0035] In accordance with another aspect of the present invention, any embodiment of the above-described embodiment of the dsRNA agent of the invention Cells containing the cells are provided. In some embodiments, the cells are mammalian cells, optionally human cells. It is a cell.

[0036] A method for inhibiting the expression of the AGT gene in cells is provided according to another aspect of the present invention. The method described above is: (i) implementing either the aforementioned dsRNA agent or the aforementioned composition of the present invention. This involves preparing cells containing an effective amount of morphology. In a particular embodiment, the method is Furthermore: (ii) Maintain the prepared cells for a sufficient amount of time to degrade the mRNA transcript of the AGT gene. This includes, in some cases, the inhibition of AGT gene expression in cells. In this embodiment, the cells are cells in the subject, and the dsRNA agent is administered subcutaneously to the subject. In some embodiments, the cells are cells in the target, and the dsRNA agent is the target. It is administered intravenously. In a particular embodiment, the method further involves administering a dsRNA agent to the subject. The method of evaluation includes assessing the inhibition of the AGT gene after administration, and the means of this evaluation are: (i) Determining the physiological characteristics of one or more AGT-related diseases or conditions in elephants; AG Baseline pre-treatment physiological characteristics of T-related disorders or conditions and / or AGT-related disorders Alternatively, by comparing the physiological characteristics of a control of the disease condition with the determined physiological characteristics. This includes, for example, that the comparison indicates whether or not the expression of the AGT gene is inhibited in the subject. In this embodiment, the determined physiological characteristic is the level of AGT in the blood. In the embodiment of the part, the determined physiological characteristics are systolic blood pressure (SBP) and diastolic blood pressure. Blood pressure, including DBP and mean arterial pressure (MAPR). Blood AGT level and / Alternatively, a decrease in blood pressure means a decrease in AGT gene expression in the subject.

[0037] A method for inhibiting AGT gene expression in a subject, according to another aspect of the present invention, The embodiment of the dsRNA formulation described above or the embodiment of the composition described above is administered to the subject in an effective amount. A method is provided which includes the following: In some embodiments, the dsRNA agent is administered subcutaneously to the target. It is administered. In certain embodiments, the dsRNA agent is administered intravenously to the subject. In terms of administration methods, the method further evaluates the inhibition of the AGT gene after administration of the dsRNA agent. This evaluation includes: (i) one AGT-related disease or condition in the subject (ii) Determining one or more physiological characteristics of AGT-related diseases or conditions Pre-treatment physiological characteristics of Sline and / or physiological characteristics of controls with AGT-related diseases or conditions This involves comparing the characteristics with the determined physiological characteristics, and the comparison is in relation to A in the subject. This includes indicating whether or not the expression of the GT gene is inhibited; in some embodiments, this determination. The physiological characteristic determined is the level of AGT in the blood; in some embodiments, The physiological characteristics identified include systolic blood pressure (SBP), diastolic blood pressure (DBP), and mean arterial pressure. Blood pressure (such as MAPR) is measured. A decrease in blood AGT levels and / or blood pressure is observed in the subject. This means a decrease in AGT gene expression.

[0038] A method for treating a disease or condition associated with AGT protein, according to another aspect of the present invention. Therefore, in order to inhibit AGT gene expression, the above-mentioned embodiment of the dsRNA agent of the present invention Any embodiment or a method comprising administering the above-mentioned composition of the present invention to a subject in an effective amount Provided. In certain embodiments, AGT-related disorders include: hypertension, high blood pressure, and borderline hypertension. Blood pressure, essential hypertension, secondary hypertension, isolated systolic or diastolic hypertension, pregnancy-related conditions. Diabetic hypertension, treatment-resistant hypertension, refractory hypertension, paroxysmal hypertension, renovascular hypertension Hypertension, Goldblatt hypertension, intraocular hypertension, glaucoma, pulmonary hypertension, portal hypertension, systemic Venous hypertension, systolic hypertension, unstable hypertension; hypertensive heart disease, hypertensive nephropathy, aterostomy Diabetic arteriosclerosis, arteriosclerosis, vascular disease, diabetic nephropathy, diabetic retinopathy, chronic heart failure, Cardiomyopathy, diabetic cardiomyopathy, glomerulosclerosis, aortic stenosis, aortic aneurysm, ventricular fibrosis, heart failure, Myocardial infarction, anguina, stroke, kidney disease, renal failure, systemic sclerosis, intrauterine growth restriction (I Selected from UGR and fetal growth restriction. In some embodiments, this method further This includes administering an additional treatment regimen to the subject. In some embodiments, the additional The treatment regimen includes the treatment of AGT-related diseases or conditions. In certain embodiments, additional The treatment regimen is: one or more AGT antisense polynucleotides of the present invention Administering to elephants; administering to non-AGT dsRNA therapeutic agents; and target odor This includes bringing about behavioral changes; in some embodiments, non-AGT dsRN A therapeutic agent may include the following: additional therapeutic agents, such as diuretics, angiotensin-converting enzyme (ACE) ) inhibitors, angiotensin II receptor antagonists, beta-blockers, vasodilators, calcium Umum channel blockers, aldosterone antagonists, α2 agonists, renin inhibitors Harmful agents, α-blockers, peripherally acting adrenergic agonists, selective D1 receptor partial agonists, Non-selective alpha-adrenergic antagonists, synthetic steroidal antimineralocorticoids, or the above Any combination of the above, and one of the antihypertensive drugs formulated as a combination of drugs. There are two.

[0039] In some embodiments, the dsRNA agent is administered subcutaneously to the subject. The dsRNA agent is administered intravenously to the subject. In some embodiments, the method is further To determine the effectiveness of double-stranded ribonucleic acid (dsRNA) agents administered to the subjects. Includes. In some embodiments, means for determining the effectiveness of treatment in a subject are: (i) (ii) Determining one or more physiological characteristics in elephants; (ii) AGT-related diseases or This involves correlating the disease state with the determined physiological characteristics, and the comparison is then applied to the subject. The effectiveness of the double-stranded ribonucleic acid (dsRNA) agent, and one of the levels of effectiveness, Including indicating one or more; in some embodiments, its determined physiological characteristics This is the level of AGT in the blood; in some embodiments, the determined physiological characteristics are Blood pressure, including systolic blood pressure (SBP), diastolic blood pressure (DBP), and mean arterial pressure (MAPR). It is pressure. A decrease in blood AGT levels and / or blood pressure is due to the administration of double-stranded ribonuclear nuclei to the subject. This indicates the effectiveness of acid (dsRNA) agents.

[0040] According to another aspect of the present invention, baseline pre-treatment levels of AGT protein in a subject A method for reducing the level of AGT protein in a subject compared to Bell, To reduce the level of T gene expression, any of the above dsRNA agent embodiments of the present invention The embodiment or any of the above-described compositions of the present invention is administered to the subject in an effective amount. A method is provided which includes the following. In some embodiments, the dsRNA agent is administered subcutaneously to the subject. Alternatively, it may be administered intravenously.

[0041] According to another aspect of the present invention, baseline treatment of AGT-related disease or condition in a subject Compared to the pre-treatment physiological characteristics, the physiological characteristics of AGT-related diseases or conditions in the subjects were improved. A method is provided for modifying the physiological characteristics of AGT-related diseases or conditions in the subject. To modify the characteristics, any embodiment of the above dsRNA agent configuration of the present invention or the present invention This includes administering an effective amount of any embodiment of the above-mentioned composition to a subject. In the administration method, the dsRNA agent is administered to the subject subcutaneously or intravenously. The physiological characteristic is the level of AGT in the blood; in some embodiments, it is determined The physiological characteristics identified include systolic blood pressure (SBP), diastolic blood pressure (DBP), and mean arterial pressure. This refers to blood pressure such as MAPR.

[0042] Array description Double strand AD00051~AD00122-19-2, AD00163-3, AV012 Table 1 shows AV01257 and AV01711, along with their sense strand sequences. vinegar.

[0043] Double strand AD00051~AD00122-19-2, AD00163-3, AV012 Table 1 shows 27~AVAV01257 and AV01711, and their antisense chains. Show the array.

[0044] Sequence ID 519 is human angiotensinogen (AGT) mRNA [see NCBI]. Array:NM_001384479.1]: [ka] [ka]

[0045] Sequence ID 520 is mouse angiotensinogen (AGT) mRNA [NCBI reference]. Reference array: NM_007428.4] [ka]

[0046] Sequence ID 521 is cynomolgus monkey angiotensinogen (AGT) mRNA [NC BI reference sequence: NM_001283634.1] [ka]

[0047] In the sequences shown in Table 2, the chemical modifications are: uppercase: 2'-fluoro; lowercase: 2'-OM e; thiophosphate: * It is shown as follows.

[0048] In the sequences shown in Table 3, the delivery molecules used in in vivo studies are This is indicated by "GLO-0" at the 3' end of each sense strand. Chemical modifications are indicated by uppercase 2'. -Fluoro; lowercase: 2'-OMe; thiophosphate: * Unlocked nucleic acid: UNA; is expressed as (Note: AD00052, AD00113-AD00260: No UNA; AD002 82-AD00301: UNA version).

[0049] In the sequences shown in Table 4, the chemical modifications are: uppercase: 2'-fluoro; lowercase: 2'-OM e; thiophosphate: * Invab is denoted as "reverse debasement". [Brief explanation of the drawing]

[0050] [Figure 1] This graph shows the serum AGT protein levels in cynomolgus monkeys after administration of AD00158-1, AD00158-2, AD00163-1, AD00159-1, and AD00300-1 at 2 mg / kg, respectively. [Figure 2] This graph shows serum AGT protein levels in cynomolgus monkeys after administration of AD00163-3 10 mg / kg. [Figure 3] This graph shows the changes in serum SBP in cynomolgus monkeys after administration of AD00163-3 10 mg / kg. [Figure 4] AD00163-3 is a graph showing the mean arterial pressure (MBP) in cynomolgus monkeys after administration of 10 mg / kg. [Figure 5] AD00163-3 is a graph showing diastolic blood pressure (DBP) in cynomolgus monkeys after administration of 10 mg / kg. [Modes for carrying out the invention]

[0051] Some embodiments of the present invention are, but are not limited to, double-stranded (ds) RNAi agents, etc. This product contains an RNAi agent that can inhibit the expression of the giotensinogen (AGT) gene. Some embodiments of the invention include a composition comprising an AGT RNAi agent, and a method of using the composition. This includes methods. The AGT RNAi agents disclosed herein are delivered to cells, such as hepatocytes. It can be attached to a delivery compound for delivery. The pharmaceutical composition of the present invention has at least one The present invention may include a dsAGT agent and a delivery compound. In some embodiments of the present invention, the delivery compound It is a GalNAc-containing delivery compound. The AGT RNAi agent delivered to the cell is AGT It is possible to inhibit gene expression, thereby affecting the AGT protein of genes in cells. The production of phosphate is reduced. Using the dsRNAi agent of the present invention, AGT-related diseases and conditions can be treated. It can be treated. Examples of such dsRNAi agents include the double-stranded AD0 shown in Table 1. Examples include 0051 to AD00122-19-2. In some embodiments, preferred Examples of dsRNAi agents include double-stranded AD00158, AD00163, and AD001 Examples include 59, AD00290, AD00300, or AD00122. Other implementations In this state, preferred dsRNAi agents include, for example, AD00158-1, AD00 158-2, AD00163-1, AD00163-3, AD00159-1 or AD0 0300-1 is one example. In some other embodiments, such dsRNAi agents These are double-stranded AD00158, AD00163, AD00163-3, AD00159, A Dual-strand variants such as variants of D00290, AD00300, or AD00122 These are some examples.

[0052] In some embodiments of the present invention, by reducing AGT expression in cells or subjects, In each cell or subject, a disease or condition associated with AGT expression is treated. Examples of diseases and conditions that can be treated by reducing activity without limitation include hypertension. Hypertension, borderline hypertension, essential hypertension, secondary hypertension, isolated systolic or diastolic hypertension, Pregnancy-related hypertension, diabetic hypertension, treatment-resistant hypertension, refractory hypertension, paroxysmal hypertension, Renal vascular hypertension, Goldblatt hypertension, intraocular hypertension, glaucoma, pulmonary hypertension, portal hypertension Systemic venous hypertension, systolic hypertension, unstable hypertension; hypertensive heart disease, hypertensive nephropathy Atherosclerosis, arteriosclerosis, vascular disease, diabetic nephropathy, diabetic retinopathy, chronic Heart failure, cardiomyopathy, diabetic cardiomyopathy, glomerulosclerosis, aortic stenosis, aortic aneurysm, ventricular fibrosis, Heart failure, myocardial infarction, anguina, stroke, kidney disease, renal failure, systemic sclerosis, intrauterine growth restriction (IUGR), and fetal growth restriction.

[0053] AGT single-stranded (ssRNA) and double-stranded (dsRNA) agents that inhibit AGT gene expression How to prepare and use compositions containing AGT gene expression, and how to produce Compositions and methods for treating, or regulating, diseases and conditions are described below. The term "RNAi" is well known in this field and may also be called "siRNA".

[0054] As used herein, the term "RNAi" includes RNA and RNA-induced It mediates targeted cleavage of RNA transcripts via the Erasing Complex (RISC) pathway. It means an agent. As is well known in the art, the RNAi target region and This includes messenger RNA (mRNA), which is the RNA-treated product of the primary transcript, and other genetic materials. This refers to the adjacent region of the nucleotide sequence of an RNA molecule formed during gene transcription. The target portion acts as a substrate for RNAi-directed cleavage at or near this portion. It is at least long enough to accomplish this. The target sequence is 8-30 nucleotides long (including), 10-30 nucleotides in length (including), 12-25 nucleotides in length (including), 15 ~23 nucleotides in length (including), 16~23 nucleotides in length (including), or 18 It can be up to 23 nucleotides in length (including), and all shorter lengths within each specified range Includes. In some embodiments of the present invention, the target sequences are 9, 10, 11, 12, 13, 1 4, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25 or 26 nuk This is the rheotide length. In certain embodiments, the target sequence is all the subranges between them. The length is 9 to 26 nucleotides, including integers. For example, the specification of the present invention Although not intended to be limited to the embodiments, the target sequences are 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 2 The RN of the AGT gene is 5, 26, 27, 28, 29, or 30 nucleotides in length. It is completely, or at least substantially, complementary to at least a portion of the transcript. Some aspects of the invention involve one or more AGT dsRNA agents and a drug-acceptable carrier. A pharmaceutical composition comprising the body and. In specific embodiments of the present invention, A described herein GT RNAi inhibits the expression of AGT proteins.

[0055] As used herein, "dsRNA agent" refers to an agent that degrades target mRNA transcripts. , or RNA or RNA-like (e.g.,) that can inhibit the translation of target mRNA transcripts. This refers to a composition containing chemically modified RNA (RNA) oligonucleotide molecules. Although we do not wish to be constrained, the dsRNA agent of the present invention operates via the RNA interference mechanism. It can act (that is, the mechanism of the RNA interference pathway in mammalian cells (RNA-induced silencing) It induces RNA interference by interacting with the RISC complex or other complexes, or alternatively, It may act through the following mechanism or pathway. In plant, invertebrate, and vertebrate cells. Methods for silencing genes are well known in this field (e.g., Shar p et al.,Genes Dev.2001,15:485;Bernstein ,et al.,(2001)Nature409:363;Nykanen,et a l., (2001) Cell 107:309; and Elbashir, et al., (2001) Genes Dev. 15:188) see), the entirety of that disclosure is referenced More incorporated herein. Gene silencing methods known in the art are AGT To achieve inhibition of expression, use in conjunction with the disclosures provided herein. It is possible.

[0056] The dsRNA agents disclosed herein consist of a sense strand and an antisense strand, and are not limited to However: Short-chain interfering RNA (siRNA), RNAi agents, microRNA (miRNA) Examples include short hairpin RNA (shRNA) and Dicer substrates. The antisense strand of the listed dsRNA agent is at least partially complementary to the targeted mRNA. The method involves targeting and inhibiting target gene expression using dsRNA double helix of various lengths. It is understood in this field that this is possible. For example, 19, 20, 21, 22, and 2 dsRNAs with a 3-base pair double-strand structure are known to effectively induce RNA interference. (Elbashiret al., EMBO 2001, 20:6877-688) 8) Shorter or longer RNA double-strand structures are also effective in inducing RNA interference. This is also known in the art. In a particular embodiment of the present invention, AGT dsRNA is , may include at least one strand having a length of at least 21 nucleotides, or the double helix may be Based on the lengths derived from at least one of the sequences shown in Tables 1-4, 1, 2, or 3 nucleotides It may have a length obtained by subtracting or less than that. Compared to the dsRNAs shown in Tables 1-4, A reduction of 4 nucleotides at one or both ends may also be effective. In some embodiments, the AGT dsRNA agent is one or more sequences from Tables 1-4. At least 15, 16, 17, 18, 19, 20 or more adjacent nucleotides from It may have a subsequence and may include the entire sequence (also referred to herein as the "parent" sequence). The level of inhibition caused by dsRNA, and 5%, 10%, 15%, 20%, 25%, and It differs in its ability to inhibit AGT gene expression at a level exceeding 30%.

[0057] Specific embodiments of the compositions and methods of the present invention include a composition comprising single-stranded RNA and / Alternatively, single-stranded RNA may be administered as the target. For example, antisense strands shown in any of Tables 1-4. It can be used as a composition or within a composition, and when administered to a subject, it can be used in the subject. This reduces AGT polypeptide activity and / or AGT gene expression. Tables 1-4 show: The antisense and sense strand cores are related to the base sequence of some AGT dsRNA agents. It indicates how to stretch it. It may be contained in certain compositions of the present invention and / or certain compositions of the present invention Single-stranded antisense molecules that can be administered by this method are referred to herein as "single-stranded antisense." It is called a "drug" or an "antisense polynucleotide agent". Contained in a specific composition of the present invention A single-stranded sense molecule that can be and / or can be administered by a specific method of the present invention is described herein as a "single-stranded sense agent" or a "sense polynucleotide agent". As used herein The term "base sequence" means a polynucleotide sequence that does not include chemical modifications or delivery compounds For example, the sense strand shown in Table 1 corresponds to the corresponding base sequence in Table 3; the corresponding sequences in Table 3 show their respective chemical modifications and delivery compounds The sequences disclosed herein can be assigned identifiers. For example, a single-stranded sense sequence can be identified by a "sense strand SS number"; a single-stranded antisense sequence can be identified by an "antisense strand AS number"; and a double-strand containing a sense strand and an antisense strand can be identified by a "double-strand AD number".

[0058] Table 1 includes sense and antisense strands and provides identification numbers for the double-strands formed by the sense and antisense strands in the same column of Table 1. In a specific embodiment of the present invention In a specific embodiment of the present invention, the antisense sequence contains the nucleobase u or the nucleobase a at its first position. In a specific embodiment of the present invention In a specific embodiment of the present invention, the antisense sequence contains the nucleobase u at its first position. As used herein, the term "matching position" means the position in each strand that "pairs" with each other in a sense when the two strands act as a double-strand For example, in a 21-nucleobase sense strand and a 21-nucleobase antisense strand, the nucleobase at the 1st position of the sense strand is at the "matching position" with the nucleobase at the 21st position of the antisense strand. For example, in a 21-nucleobase sense strand and a 21-nucleobase antisense strand, the nucleobase at the 1st position of the sense strand is at the "matching position" with the nucleobase at the 21st position of the antisense strand. For example, in a 21-nucleobase sense strand and a 21-nucleobase antisense strand, the nucleobase at the 1st position of the sense strand is at the "matching position" with the nucleobase at the 21st position of the antisense strand. ​In another non-restrictive example in 23 nucleic acid base sense strands and 23 nucleic acid base antisense strands, Therefore, the nucleic acid base at position 2 of the sense strand is in a matching position with position 22 of the antisense strand. In another non-restrictive example in 18 nucleic acid base sense strands and 18 nucleic acid base antisense strands, Therefore, the nucleic acid base at position 1 of the sense strand is in a matching position with position 18 of the antisense strand. The nucleic acid base at position 4 of the sense strand matches the nucleic acid base at position 15 of the antisense strand. It is in the position. Those skilled in the art will know the difference between the sense and antisense chains of double chains and paired chains. It should be clear how the matching location is identified.

[0059] The last column in Table 1 contains the dual sense and antisense arrays in the same column of the table. The double-strand AD / AV numbers of the chain are shown. For example, Table 1 shows the corresponding sense and antisense numbers. We disclose a double helix containing a double helix sequence, named "Double Helix AD Number AD00051". Therefore, each column in Table 1 contains the sense and antisense arrays shown in the same column. The identifiers that identify the double helix of the present invention and that are named for each double helix are shown at the end of the column in the field. It will be done.

[0060] Some embodiments of the method of the present invention include R containing the polynucleotide sequence shown in Table 1. A NAi agent is administered to the subject. In some embodiments of the present invention, an RN is administered to the subject. The AI ​​agent contains at least one of the base sequences shown in Table 1, and is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 1 The present invention includes a double helix containing sequence modifications 9, 20, 21, 22, 23, or 24. In some embodiments of the method, RNAi agents with polynucleotide sequences shown in Table 1 are delivered by a delivery molecule. further includes linking thereto, and non-limiting examples thereof are delivery compounds containing GalNAc exist.

[0061] [Table 1]

[0062] [Table 2]

[0063] [Table 3]

[0064] [Table 4]

[0065] [Table 5]

[0066] [Table 6]

[0067] [Table 7]

[0068] [Table 8]

[0069] [Table 9]

[0070] [Table 10]

[0071] Table 2 shows the antisense and sense strand sequences of specific chemically modified AGT RNAi agents of the present invention. This shows that, in some embodiments of the present invention, R has the polynucleotide sequence shown in Table 2. The NAi agent is administered to cells and / or subjects. In some embodiments of the present invention, see Table 2. An RNAi agent having the polynucleotide sequence described above is administered to the target. In this embodiment, the RNAi agent administered to the subject comprises the double hemisphere shown in the first column of Table 2. Furthermore, the sense and antisense shown in the third and sixth columns of the same column in Table 2, respectively. This includes sequence modification of the lance chain sequence. In some embodiments of the present invention, the sequences shown in Table 2 are paired Compounds capable of delivering RNAi agents to elephant cells and / or tissues (as specified herein) In certain embodiments of the present invention, it is possible to (also called conjugate). A non-restrictive example of a useful delivery compound is a GalNAc-containing compound. In Table 2, The first column represents the double-stranded AD number or AV number of the base sequence corresponding to Table 1. With respect to the base sequence identified by the D number, the sense and antisense strands are contained Not only is the base sequence shown, but the specified chemical modifications are also shown in the same column of Table 2. For example For example, the first column of Table 1 shows the double helix, identified as double helix AD number AD00051, and the double helix together form In contrast to the single-stranded sense and antisense base sequences shown in Table 2, the double-stranded sequence is shown. As a double strand in AD number AD00051, it is AD00051-SS and AD0 Includes the base sequence of 0051-AS, and the sense and A shown in the third and sixth columns respectively. The sense sequence contains chemical modifications. See the "Sense Chain SS Number" in column 2 of Table 2. " is the identifier for the sense array (including modifiers) shown in the third column of the same column. The "Antisense Chain AS Number" in column 5 refers to the antisense sequence (including modifications) shown in column 6. This is the identifier for the specified (mu).

[0072] [Table 11]

[0073] [Table 12]

[0074] [Table 13]

[0075] [Table 14]

[0076] [Table 15]

[0077] Table 3 shows the antisense and sense strand sequences of specific chemically modified AGT RNAi agents of the present invention. This shows that in some embodiments of the present invention, the RNAi agents shown in Table 3 are used on cells and / or target cells. It is administered to. In some embodiments of the present invention, the polynucleotide sequences listed in Table 3 are used The RNAi agent is administered to the subject. In some embodiments of the present invention, the agent is administered to the subject. The RNAi agents include the double hemispheres identified in column 1 of Table 3, and the same column in Table 3 Sequence modifications and / or shown in the sense and antisense chain sequences of the 3rd and 6th columns, respectively. It contains a delivery compound. Its sequence is as described herein for specific in vivo (in v ivo) Used in experimental research. In some embodiments of the present invention, the sequences shown in Table 3 are The compound for delivery is linked (also referred to as conjugated herein). A non-restrictive example of such a compound is a GalNAc-containing compound, namely "GLX- The delivery compound identified as "n" is located on the sense chain in column 3 of Table 3. In use, "GLX" refers to a "GLS" or "GLO" delivery compound (where "X" is "S" or It is used to mean (which can be "O"), and GLX-n is a synthesis process Either of the following can be linked to the 3' or 5' end of the oligonucleotide of the present invention. These may be GLS and GLO. As an unrestrictive example, GLX-13 and GLX-14 are, During the synthesis process, it can be linked to the 3' end of the oligonucleotide of the present invention, GLX -5 and GLX-15 are linked to the 5' end of the oligonucleotide of the present invention during the synthesis process. It can be concluded. In some embodiments, as used herein and shown in Table 3 Furthermore, "GLX-n" is used to represent a compound containing a bound GalNAc, and Compounds GLS-1, GLS-2, GLS-3, GLS-4, GLS-5, GLS-6, G LS-7, GLS-8, GLS-9, GLS-10, GLS-11, GLS-12, GL S-13, GLS-14, GLS-15, GLS-16, GLO-1.GLO-2, GL O-3, GLO-4, GLO-5, GLO-6, GLO-7, GLO-8, GLO-9, GLO-10, GLO-11, GLO-12, GLO-13, GLO-14, GLO-1 It is either 5 or GLO-16. In some implementations, but not limited to, all This is described herein by Jayaprakash, et al., (2014) G, useful for ligation, disclosed in J.Am.Chem.Soc., 136, 16958 GLO-0, containing alNAc, is disclosed in prior art as being usable for ligation. This represents a GalNAc-containing compound. In some embodiments, those skilled in the art will know that it is not limited to... :GLS-1, GLS-2, GLS-3, GLS-4, GLS-5, GLS-6, G LS-7, GLS-8, GLS-9, GLS-10, GLS-11, GLS-12, GL S-13, GLS-14, GLS-15, GLS-16, GLO-1, GLO-2, GL O-3, GLO-4, GLO-5, GLO-6, GLO-7, GLO-8, GLO-9, GLO-10, GLO-11, GLO-12, GLO-13, GLO-14, GLO-1 The present invention, together with a conjugated delivery compound containing one of 5 and GLO-16, RNA compounds can be prepared and used. The structure of each of these compounds is described in this specification. Provided elsewhere in the book. Column 1 of Table 3 shows the sense and antisense in that column of the table. Provides the double-strand AD number of the double-stranded sequence assigned to the sequence. For example, double-strand AD number A D00052 consists of the sense chain AD00052-SS and the antisense chain AD00052-A It is a double helix composed of S. Each column in Table 3 represents one sense strand and one antisense strand. We provide and disclose a double helix composed of a sense strand and an antisense strand. The "Sense Chain SS Number" in column 2 of Table 3 corresponds to the sense sequence (modified) shown in column 3 of the same column. This is the identifier for the designation of (including). The "Antisense Chain AS Number" in column 5 of Table 3. " is the identifier for the antisense sequence (including modifications) shown in column 6. The identifier for the linked GalNAc-containing GLO compound is indicated as GLO-0, and also as GLO-n. Another GLS-n compound may be substituted in place of the compound designated as GLO-0. The compounds obtained are also included in embodiments of the methods and / or compositions of the present invention. Please understand this.

[0078] [Table 16]

[0079] [Table 17]

[0080] [Table 18]

[0081] [Table 19]

[0082] [Table 20]

[0083] [Table 21]

[0084] Table 4 shows the antisense and sense strand sequences of specific chemically modified AGT RNAi agents of the present invention. This shows that, in some embodiments of the present invention, R has the polynucleotide sequence shown in Table 4. A NAi agent is administered to the subject. In some embodiments of the present invention, an RN is administered to the subject. The AI ​​agent contains a double helix identified in column 1 of Table 4, and in columns 3 and 6 of Table 4. Sequence modifications and / or delivery compounds indicated by the same sense and antisense chain sequences in the same column Includes. In some embodiments of the present invention, the sequences shown in Table 4 are used to deliver RNAi agents to target cells. It can be linked to a compound that can be delivered to and / or tissue. A non-limiting example of a useful delivery compound in this embodiment is a GalNAc-containing compound. In Table 4, the term "GLX-n" refers to the presence of GalNAc in the sense strand shown. It refers to the compounds contained. For example, the terms "GLO-0" and "GLS-5" refer to the compounds contained. Each represents a different GalNAc-containing compound bonded to the sense chain. GLO-0 indicates... The compound may be substituted with another compound, either GLO-n or GLS-n. Furthermore, it is understood that the resulting compounds are also included in the methods and / or compositions of the present invention. Please do so. Similarly, compounds designated as GLS-5 are also GLS-n or GLO-n compounds. The compound that can be substituted by another substance and obtained is the method and / or combination of the present invention. This is included in the embodiments of the product. Table 4 shows the bound GalNAC-containing compounds. The compound GLX-n used is the same as compounds GLS-1, GLS-2, GLS-3, G LS-4, GLS-5, GLS-6, GLS-7, GLS-8.GLS-9, GLS-1 0, GLS-11, GLS-12, GLS-13, GLS-14, GLS-15, GLS -16, GLO-1, GLO-2, GLO-3, GLO-4, GLO-5, GLO-6, GLO-7, GLO-8, GLO-9, GLO-10, GLO-11, GLO-12, G These are LO-13, GLO-14, GLO-15, and GLO-16, and their respective structures This is provided elsewhere in this specification. Column 1 of Table 4 corresponds to the double helix shown in Table 3. This represents the double-stranded AD number. The double-stranded AD number identifies the corresponding double-stranded sequence in Table 3. The sense, antisense, and double-stranded sequences in 4 correspond to the same double-stranded AD numbers in Table 3. The base sequence is identical to the one shown in Table 4, but the sequences and double helix in Table 4 correspond to the sequences shown in Table 3. This indicates that the sequence and double hemisphere have different chemical modifications and / or delivery compounds compared to the original. For example, as shown in Table 4, sequences AD00113-1-SS and AD00113-1-A S and its double-strand AD number AD00113-1 are shown in Table 3 as AD00113- SS (Sense), AD00113-AS (Antisense), and double-stranded AD number AD001 It has the same base sequence as 13, but has chemical modifications and / or delivery compounds as shown in each table. Table 4, column 1 identifies the double-stranded AD number; each column is identified by number. The double helix includes the sense and antisense helix shown in the third and sixth columns, respectively, in the same column. Furthermore, it includes modifications, each of which is a GLO- or G attached to the 3' or 5' end of the sense strand. It contains an LS-delivery compound.

[0085] [Table 22]

[0086] [Table 23]

[0087] [Table 24]

[0088] [Table 25]

[0089] [Table 26]

[0090] [Table 27]

[0091] [Table 28]

[0092] [Table 29]

[0093] mismatch In particular, when the mismatch is located within the terminal region of the dsRNA, the mismatch affects the effectiveness of the dsRNA. It is known to those skilled in the art that it can withstand force. Certain mismatches, such as smatches, are tolerated better (Duet al., A syste matic analysis of the silencing effects of an active siRNA at all single-nucleots ide mismatched target sites.Nucleic Acid s Res.2005 Mar21;33(5):1671-7.Doi:10.109 3 / nar / gki312.Nucleic Acids Res.2005;33(1 1):3698). In some embodiments of the methods and compounds of the present invention, AGT dsR NA agents may contain one or more mismatches with respect to the AGT target sequence. In terms of form, the AGT dsRNA agent of the present invention does not contain mismatches. Specific implementation In terms of form, the AGT dsRNA agent of the present invention contains one or fewer mismatches. In one embodiment, the AGT dsRNA agent of the present invention contains 2 or fewer mismatches. In certain embodiments, the AGT dsRNA agent of the present invention has three or fewer mismatches. Contains. In some embodiments of the present invention, the antisense strand of the AGT dsRNA agent is It contains mismatches for AGT target sequences that are not located in the center of the complementary region. In this embodiment, the antisense strand of the AGT dsRNA agent is located at the 5' or 5' of the complementary region. It is located within the last 5, 4, 3, 2, or 1 nucleotide from one or both of the 3' ends. It contains 2, 3, 4 or more mismatches against the AGT target sequence. To determine whether an AGT dsRNA agent containing ci is effective in inhibiting AGT gene expression. To do so, the methods described herein and / or methods known in the art may be used. Cut.

[0094] Complementarity Unless otherwise specified, the term “complementarity” as used herein This is the second nucleotide sequence (for example, AGT dsRNA antisense strand or single-stranded A The first nucleotide sequence (e.g., AGT ds) for nucleotide polynucleotides When used to describe the relationship between RNA drug sense strands or targeted AGT mRNA) , hybridizes with oligonucleotides containing a second nucleotide sequence, and under specific conditions Below, it forms a double helix or double-strand structure [under the physiological conditions of mammals (or outside of the body)]. [Under similar conditions], it forms hydrogen bonds between base pairs, including the first nucleotide sequence. This refers to the capabilities of oligonucleotides or polynucleotides. Other conditions may also apply, such as rationally related conditions. Hybridized Nucleotide The conditions best suited for testing the complementarity of two sequences based on the final application of the method. Those skilled in the art will be able to determine the pair. The complementary sequence is the Watson-Crick base pair. Or containing non-Watson-Crick base pairs and being a necessary condition for the above hybridization Natural or modified nucleotides or nucleotide ears to the extent that at least the following conditions are met Includes bucks. Sequence identity or complementarity does not depend on modification.

[0095] For example, the complementary sequences within the AGT dsRNA described herein may be one or both An oligonucleotide containing the second nucleotide sequence throughout the entire length of the nucleotide sequence For a nucleotide or polynucleotide, an oligonucleotide containing the first nucleotide sequence or Contains base pairs of polynucleotides. Such sequences are "completely complementary" to each other as defined herein. It can be called when two oligonucleotides hybridize into one In embodiments where it is designed to form multiple single-chain overhangs, Overhangs are considered mismatches determined in this specification based on complementarity. It should be understood that one oligonucleotide has a length of 19 nucleotides. AGT d contains tide and another oligonucleotide with a length of 20 nucleotides. In sRNA agents, long oligonucleotides are completely different to short oligonucleotides. AGT dsRNA agents containing a 19-nucleotide sequence complementary to the above are described herein. For the purposes of this, it can be called "completely complementary". Therefore, as used herein, "Completely complementary" means all bases (100%) in the adjacent sequence of the first polynucleotide. However, hybridizing with the same number of bases in the adjacent sequence of the second polynucleotide This means that the adjacent sequence may contain all or part of the first or second nucleotide sequence.

[0096] As used herein, the term “substantially complementary” refers to a hybrid of nucleic acid base sequences. In a pair, at least about 85% of the bases in the adjacent sequence of the first polynucleotide ( (Not all) but the same number of bases in the adjacent sequence of the second polynucleotide and hybridized It means to hybridize. When hybridized, the two sequences become one or more micronized. Smatch base pairs, for example, containing at least 1, 2, 3, 4, or 5 mismatch base pairs In such cases, the term "substantially complementary" means that, for example, its end use is via a RISC pathway. It retains the ability to hybridize under conditions most relevant to inhibiting AGT gene expression. However, relative to the second sequence, the first sequence forms 15, 16, 17, 18, 19, 2 0, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 base pairs (bp) The term "partially complementary" can be used in relation to double hemispheres up to the first polynucleate. At least 75% (but not all) of the bases in the adjacent sequence of ocide are second polynuclei Nucleic acid base sequence hybridization occurs when a rheotide hybridizes with the same number of bases in its adjacent sequence. In this specification, it may be used to mean a reduced pair. In some embodiments "Partially complementary" means that at least one of the bases in the adjacent sequence of the first polynucleotide is 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% are the same in the adjacent sequence of the second polynucleotide. It means hybridizing with a number of bases.

[0097] "Complementary," "fully complementary," "substantially complementary," and "partially complementary" are AG T dsRNA agent sense strand and antisense strand, AGT dsRNA agent antisense A single-stranded antisense oligonucleotide pair with the sequence of the target AGT mRNA and It can be used to indicate a base mismatch with the target AGT mRNA sequence. The "antisense strand of GT dsRNA agent" is the same as the "AGT antisense polynucleotide agent". Please understand that these can mean the same sequence.

[0098] In this specification, when referring to nucleic acid sequences, the terms "substantially identical" or "substantial" are used. The term "identity" refers to a nucleic acid sequence that is at least approximately 85% identical to a reference sequence. Uniformity, preferably at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, This means that it contains sequences with at least 98% or at least 99% sequence identity. The percentage of sequence identity is calculated by comparing two sequences that are optimally aligned across the comparison window. It is determined by comparing the columns. The percentage is determined by the number of identical nucleic acid bases in both columns. Determine the number of positions in the column, obtain the number of matched positions, and then use the total number of positions in the comparison window. By dividing the number of match positions and multiplying the result by 100, you can obtain the percentage of sequence identity. It is calculated by [formula]. The present invention disclosed herein is described herein (for example, in Tables 1-5). It includes a nucleotide sequence that is substantially identical to the shown sequence. In some embodiments, The nucleotide sequence is identical to, or at least identical to, the sequences disclosed herein (e.g., Tables 1-4). Approximately 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94% They are 95%, 96%, 97%, 98%, or 99% identical.

[0099] As used herein, the term “sequence-containing chain” refers to a chain that does not conform to standard nucleotide nomenclature. Oligonucleotides containing a chain of nucleotides described by a sequence referenced using The term "double-stranded RNA" or "dsRNA" as used herein means. "Sense" and "antisense" orientation for RNA molecules or target AGT RNA. It contains two antiparallel and substantially or completely complementary nucleic acid chains, which are referred to as having a characteristic. This refers to a sequence containing a complex of RNAi molecules having a hybridized double-stranded region. The double-stranded region enables the specific degradation of target AGT RNA via the RISC pathway. It can be any desired length, but is typically 9 to 30 base pairs long, for example, 15 to 30 base pairs long. This is true. Considering a double helix of 9 to 30 base pairs, the length of that double helix can be any length within this range. For example, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29 or 30 base pairs, and among them Within a partial range, not limited to, but 15-30 base pairs, 15-26 base pairs; 15-23 base pairs , 15-22 base pairs, 15-21 base pairs, 15-20 base pairs, 15-19 base pairs, 15- 18 base pairs, 15-17 base pairs, 18-30 base pairs, 18-26 base pairs, 18-23 base pairs Pairs, 18-22 base pairs, 18-21 base pairs, 18-20 base pairs, 19-30 base pairs, 19 ~26 base pairs, 19-23 base pairs, 19-22 base pairs, 19-21 base pairs, 19-20 base pairs Base pair, 20-30 base pairs, 20-26 base pairs, 20-25 base pairs, 20-24 base pairs, 2 0-23 base pairs, 20-22 base pairs, 20-21 base pairs, 21-30 base pairs, 21-26 base pairs, 21-25 base pairs, 21-24 base pairs, 21-23 base pairs, or 21-22 base pairs They can be of paired length. They are produced in cells by treatment with dicers and similar enzymes. AGT dsRNA reagents are typically in the range of 19–22 base pairs in length. One strand of the double-stranded region of agent A is substantially complementary to the region of the target AGT RNA. It contains a sequence. The two strands forming the double-stranded structure have at least one self-complementary region. It may arise from a single RNA molecule, or it may be formed from two or more other RNA molecules. When the heavy chain region is formed from a single molecule, the molecule is the 3' end of a single strand of nucleotide. It has a double-strand structure formed between one strand and another strand at the corresponding 5' end (Honmei (Referred to as "hairpin loop" in the details). In some embodiments of the present invention, hair The pin arrangement is at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 1 Contains 3, 14, 15, 16, 17, 18, 19, 20 or more unpaired nucleotides If the two substantially complementary strands of the AGT dsRNA agent consist of separate RNA molecules... These molecules do not need to be covalently bonded, but they can be. Two chains, hairpin When covalent bonds are formed by means other than loops, the linked structure is called a "linker." The term "siRNA" also means the dsRNA agents described herein. It is used.

[0100] In some embodiments of the present invention, the AGT dsRNA agent is one of the dsRNA agents. sense and antisense, which have unpaired nucleotides or nucleotide analogs at both ends. It may contain sequences. Ends that do not contain unpaired nucleotides are called "blunt ends," and also contain nucleotides. It does not have a rheotide overhang. If both ends of the dsRNA agent are blunt, ds RNA is said to have a "blunt end". In some embodiments of the present invention, the dsRNA agent If the first end is blunt, in some embodiments the second end of the dsRNA agent is blunt. In certain embodiments of the present invention, both ends of the AGT dsRNA agent are blunt. ru.

[0101] In some embodiments of the dsRNA agent of the present invention, the dsRNA contains one or two flat There is no smooth end. In this case, there is at least one unpaired nucleo at the end of the dsRNA drug strand. There is a limit. For example, the 3' end of one strand of dsRNA extends beyond the 5' end of the other strand. A nucleotide overhang exists when it extends in both directions, or vice versa. RNA contains at least 1, 2, 3, 4, 5, 6 or more nucleotides. It may contain hangs. Nucleotide overhangs are deoxynucleotides / nucleos It may contain or consist of nucleotide / nucleoside analogs such as sides. In one embodiment, the nucleotide overhang is on the sense strand of the dsRNA agent, dsRN Located on the antisense strand of agent A, or at the ends of both dsRNA agents, and with an overhang Nucleotides are located at the 5' end or 3' end of either the antisense or sense strand of dsRNA. It should be understood that this may be present at one or both ends. In certain embodiments of the present invention And one or more nucleotides in the overhang are nucleoside thiophosphates. It can be replaced with.

[0102] As used herein, the terms “antisense strand” or “guide strand” refer to the AGT mark. This refers to a strand of AGT dsRNA agent containing a region substantially complementary to the target sequence. The terms "sense chain" or "passenger chain" used in this document are AGT ds AGT ds contains a region substantially complementary to the antisense strand region of the RNA agent. This refers to the RNA chain.

[0103] qualification In some embodiments of the present invention, the RNA of the AGT RNAi agent is chemically modified, Qualitative improvements and / or one or more other beneficial properties are obtained. The nucleic acids in the application form are synthesized and / or modified by methods well known in the art. This can be done, for example, as incorporated herein by reference, “Current pr otocols in Nucleic Acid Chemistry”,Beauc age, SL et al. (Eds.), John Wiley & Sons, In See c., New York, NY, USA. The AGT dsRNA agent of the present invention Modifications that may exist in a particular embodiment include, for example: (a) terminal modifications, e.g., 5' terminal modifications. Modifications (phosphorylation, conjugation, inverse bonding, etc.), 3' end modifications (conjugation, DNA nucleotides, inverse bonding) (b) Base modification, e.g., stabilizing base, base with expanded partner pool Alternatively, base pairing, missing base (debasalized nucleotide), or conjugated Substitutions of a base; (c) sugar modification (e.g., at the 2' or 4' position) or sugar substitution; and ( d) Modification of the main chain, such as modification or substitution of phosphate diester bonds. GT dsRNA agent, AGT antisense polynucleotide, and AGT sense polynucleotide Specific examples of RNA compounds useful for certain embodiments of rheotide are limited to: However, examples include RNA that contains a modified backbone or lacks natural nucleoside bonds. As a limited example, RNA with main chain modifications does not possess a phosphorus atom in its main chain. RNA that does not have a phosphorus atom in the interosidal backbone can be called an oligonucleoside. In a particular embodiment, the modified RNA has a phosphorus atom in its internucleoside backbone. .

[0104] The terms "RNA molecule" or "RNA" or "ribonucleic acid molecule" are used in natural expression or observed in In addition to the RNA molecules released, ribonucleotides described herein or known in the art are also present. It is understood that this includes RNA analogs and derivatives, including ocides / molecular analogs or derivatives. The terms "molecule" and "ribonucleotide" may be used interchangeably herein. RNA molecules have a nucleic acid base structure or a ribose-phosphate main chain structure (for example, as described below). Molecules that can be modified in (and contain molecular analogs or derivatives) form a double chain. It must retain the ability to do so. As a non-restrictive example, RNA molecules also have at least the ability to do so. Another modified molecule, for example, a locked nucleoside, a debased nucleoside, etc. Oside, 2'-deoxy-2'-fluoromodified nucleoside, 2'-aminomodified nucleo sides, 2'-alkyl-modified nucleosides, morpholino nucleosides, phosphoramidates Alternatively, it may include non-natural base-containing nucleosides, or combinations thereof. Part of the present invention In this embodiment, the RNA molecule has the following number of modified ribonucleosides: at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, Ribonucleosides of AGT dsRNA drug molecules up to 18, 19, 20, or full length Includes. With respect to each of the numerous modified ribonucleosides in such RNA molecules, modification They don't need to be the same.

[0105] In some embodiments, the dsRNA agent of the present invention, AGT antisense polynucleotide The , and / or AGT sense polynucleotides are selected one or more independently. Modified nucleotides and / or one or more independently selected non-phosphate diste compounds May include a nucleotide bond. Modified nucleotides and non-phosphate diester bonds as used herein. When used to mean elements that are selected, such as, "independently selected" The words do not need to have two or more selected elements that are identical to each other, but they are identical to each other. It means to obtain. The terms "nucleotide base" and "nucleotide" as used herein are used in this specification. Alternatively, "nucleic acid base" refers to heterocyclic pyrimidines or plycos, which are the standard building blocks of all nucleic acids. It is a compound, and the bases that form nucleotides are adenine (a), guanine (g), and cytonucleotides. It includes syn (c), thymine (t), and uracil (u). Nucleic acid bases are further limited to these. Although not intended to be a universal base, hydrophobic base, promiscuous base It can be modified to include cuous bases, size-extended bases, and fluorinated bases. In this specification, the terms "ribonucleotide" or "nucleotide" refer to unmodified nucleotides. It can be used to mean octide, modified nucleotide, or another site. Such substitution Significantly altering the base pair properties of an oligonucleotide containing a nucleotide that maintains a position. Instead, guanine, cytosine, adenine, and uracil can be substituted by other sites. Those skilled in the art will recognize this.

[0106] In one embodiment, a modified RN intended for use in the methods and compositions described herein A forms the desired double-strand structure and enables specific degradation of the target via the RISC pathway. A peptide nucleic acid (PNA) having the ability to make or mediate. Specific embodiment of the present invention Morphologically, AGT RNA interferants interact with the target AGT RNA sequence, thereby targeting the target It contains single-stranded RNA that instructs the cleavage of AGT RNA.

[0107] Modified RNA backbone can be, for example, phosphorothioates, chiral phosphorothioates, or phosphorothioates. Logithioate, phosphotriester, aminoalkylphosphotriester, methyl and Other alkylphosphonates (such as 3'-alkylenephosphonates and chiralphosphonates) ), phosphinate, phosphoramidate (3'-aminophosphoramidate and amino Alkyl phosphoramidates, thiophosphoamidates, thioalkylphosphonates , thioalkyl phosphotriesters, and boranophosphates (usually 3'-5' bonded) , and also having these 2'-5' bond analogues, and adjacent pairs of nucleoside units are 3' Concatenated in the form of -5' or 5'-3' or 2'-5' or 5'-2', with opposite polarity It may contain (having). Various salts, mixed salts and free acid forms are also included. Phosphorus-containing bonds The method of formation is commonly practiced in the art, and using such a method, the present invention Specific modified AGT dsRNA agents, specific modified AGT antisense polynucleotides, And / or specific modified AGT sense polynucleotides can be prepared.

[0108] Modified RNA backbone that does not contain phosphorus atoms is short alkyl or cycloalkyl nucleo Osid-to-osidic bonds, mixed heteroatoms, and alkyl or cycloalkylnucleoside bonds or formed by one or more short-chain heteroatoms or heterocyclic nucleoside bonds It has a structure. Morpholino bond (partially formed from the sugar portion of the nucleoside); Xane backbone; sulfide, sulfoxide and sulfone backbone; methylacetyl and thiomethic acid methylacetyl main chain; methylenemethylacetyl and thiomethylacetyl main chain; alkene-containing main chain Chains; sulfamate backbone; methyleneimino and methylenehydrazino backbone; sulfonates and Sulfonamide main chain; amide main chain; and other having mixed N, O, S and CH2 components Examples include those having a portion. A method for producing a modified RNA backbone that does not contain a phosphorus atom. This is commonly practiced in the art, and using such methods, the specific modified AGT of the present invention dsRNA agents, specific modified AGT antisense polynucleotides, and / or specific modified Decorated AGT sense polynucleotides can be prepared.

[0109] In certain embodiments of the present invention, but not limited to, for example, nucleotide units of sugars By using a new group instead of the nucleoside bond (i.e., the main chain), RNA mimetic The AGT dsRNA, AGT antisense polynucleotide, and / or AGT sec It is included in the nucleotide. In this embodiment, the base unit is an appropriate AGT. Maintained for hybridization with nucleic acid target compounds. Excellent hybridization Such oligomer compounds, RNA mimetic compounds, are known to have symmetry properties. This is called peptide nucleic acid (PNA). In PNA compounds, the sugar backbone of RNA is The nucleic acid bases are substituted by an amide-containing backbone, specifically an aminoethylglycine backbone. RNA mime Methods for manufacturing ticks are commonly practiced in the art, and using such methods, The specific modified AGT dsRNA agent of the present invention can be produced.

[0110] Some embodiments of the present invention relate RNA having a phosphorothioate backbone, and heteroatoms Main chain, especially --CH2--NH--CH2-, --CH2--N(CH3)--O--C H2--[known as methylene (methylimino) or MMI backbone],--CH2-- O--N(CH3)--CH2--, --CH2--N(CH3)--N(CH3)-- CH2-- and --N(CH3)--CH2----[Natural phosphodiester back chains are-- It contains oligonucleotides having the structure O--P--O--CH2--. RNA having a thioate backbone and oligonucleotides having a heteroatom backbone are produced. The methods of making are commonly practiced in the art, and using such methods, specific modified AGT dsRNA agents, specific AGT antisense polynucleotides and / or specific AGT sense polynucleotides can be manufactured.

[0111] Modified RNAs may also contain one or more substituted sugar moieties. The AGT dsRNA of the present invention , AGT antisense polynucleotides and / or AGT sense polynucleotides have 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-a lkyl; and may include one of these, where alkyl, alkenyl and alkynyl are substituted or unsubstituted C1-C 10 alkyl or C2-C 10 alkenyl and alkynyl. Exemplary suitable modifications are: O[(CH2) n O] m CH3, O(CH2) n OCH 3, O(CH2) n NH2, O(CH2) n CH3, O(CH2) n ONH2, and O( CH2) n ON[(CH2) n CH3)]2 (n and m are from 1 to about 10). Other In embodiments, the dsRNA has the following at the 2' position: C1-C 10 lower alkyl, substituted lower alkyl, aralkyl, aralkyl, O-aralkyl or O-aralkyl, S H, SCH3, OCN, Cl, Br, CN, CF3, OCF3, SOCH3, SO2CH 3, ONO2, NO2, N3, NH2, heterocycloalkyl, heterocycloaralkyl aminoalkylamino, polyalkylamino, substituted silyl, RNA cleavage group, reporter The pharmacokinetics of intercalators and AGT dsRNA agents. A base used to improve physiological properties, or AGT dsRNA agent, AGT anti Improving the pharmacodynamic properties of sense polynucleotides and / or AGT sense polynucleotides. A group used for that purpose, and other groups having similar properties; including one of them. In embodiments, the modification is 2'-methoxyethoxy(2'-O-(2-methoxyethyl ) or also known as 2'-MOE (2'-O--CH2CH2OCH3)(Marti net al., Helv. Chim. Acta, 1995, 78:486-504), In other words, it contains an alkoxy-alkoxy group. Another exemplary modification is 2'-dimethylaminoester. Toxyethoxy, also known as 2'-DMAOE as described in the following examples, O( CH2)2ON(CH3)2 groups, and 2'-dimethylaminoethoxyethoxy(2'-O Also known in the art as dimethylaminoethoxyethyl or 2'-DMAEOE ), that is, 2'-O--CH2-O--CH2--N(CH2)2. However, methods for producing modified RNA are commonly practiced in the art, and such methods are used This allows for the production of specific modified AGT dsRNA agents according to the present invention.

[0112] Other modifications include 2'-methoxy(2'-OCH3), 2'-aminopropoxy(2 Examples include '-OCH2CH2CH2NH2' and 2'-fluoro(2'-F). Similar The modification of the present invention relates to the AGT dsRNA agent, AGT antisense polynucleotide, and AG. Other positions on RNA of T sense polynucleotides and / or AGT sense polynucleotides In particular, the 3' position of the sugar on the 3' terminal nucleotide, or the 2'-5' ligated AGT dsR NA, AGT antisense polynucleotide, or AGT sense polynucleotide It can also be added at the 3' position and the 5' position of the 5' terminal nucleotide. AGT dsRNA agents, AGT antisense polynucleotides, and / or AGT sense polynucleotides Cleotides also contain sugar mimetics such as cyclobutyl moieties instead of pentofuranosyl sugars. It is possible. Methods for producing modified RNA, such as those described, are commonly practiced in this art. The present invention provides a specific modified AGT dsRNA agent, AGT AN The production of thisense polynucleotides and / or AGT sense polynucleotides can.

[0113] In some embodiments, AGT dsRNA agents and AGT antisense polynucleotides are used. D, and / or AGT sense polynucleotides are nucleic acid bases (commonly referred to as "bases" in the art). This may include modifications or substitutions (referred to as "unmodified" or "natural"). As used herein, "unmodified" or "natural" may refer to modifications or substitutions. Nucleic acid bases include the purine bases adenine (A) and guanine (G), and pyrimidine salts. Examples of modified nucleic acid bases include thymine (T), cytosine (C), and uracil (U). For other synthetic and natural nucleic acid bases, such as 5-methylcytosine (5-me-C) and 5-Hynoside, Droxymethylcytosine, xanthine, hypoxanthine, 2-aminoadenine, adenine and 6-methyl and other alkyl derivatives of guanine, adenine and 2-propyl of guanine Alkyl and other alkyl derivatives, 2-thiouracil, 2-thiothymine and 2-thiositosan ( thiocytosan) pyrimidine, 5-halouracil and cytosine, 5-propynyl Uracil and cytosine, 6-azouracil, cytosine and thymine, 5-uracil (pseudouracil) Sil), 4-thiouracil, 8-halo, 8-amino, 8-thiol, 8-thioalkyl, 8-hydroxyl and other 8-substituted adenines and guanines, 5-halo, especially 5-bromo, 5-trifluoromethyl and other 5-substituted uracils and cytosines; 7-methylguanine and 7-methyladenine, 8-azaguanine and 8-azaadenine, 7-azaguanine and Examples include 7-azaadenine, as well as 3-azaguanine and 3-azaadenine. Additional nucleic acid bases that may be included in certain embodiments of the AGT dsRNA agent are, in the Art of It is publicly known, for example: Modified Nucleosides in Biochemistry mistry,Biotechnology and Medicine,Herdew ijn,P.Ed.Wiley-VCH2008;The Concise Encyc lopedia Of Polymer Science And Engineeri ng,pages 858-859,Kroschwitz,JL,Ed.John Wiley & Sons,1990,English et al.,Angewan dte Chemie,International Edition,1991,30 ,613,Sanghvi,Y S.,Chapter15,dsRNA Research ch and Applications, pages 289-302, and Crook See e, ST and Lebleu, B., Ed., CRC Press, 1993. Please include dsRNA, including those described herein, which include nucleic acid base modifications and / or substitutions. , AGT antisense strand polynucleotide, and / or AGT sense strand polynucleotide Methods for producing are commonly practiced in the art, and using such methods, the present invention Specific modified AGT dsRNA agents, AGT sense polynucleotides and / or AGT agents It is possible to manufacture cysens polynucleotides.

[0114] The present invention relates to AGT dsRNA agents, AGT antisense polynucleotides, and / or A A specific embodiment of GT sense polynucleotides is one or more LOK nucleic acids (LNAs) It contains RNA modified to include the following: The lock nucleic acid has additional 2' and 4' carbons linked together. This nucleotide has a modified ribose site that includes a crosslink. This structure is the 3' terminal structure Effectively "locks" ribose in conformation. The AGT dsR of the present invention NA agent, AGT antisense polynucleotide and / or AGT sense polynucleotide By adding lock nucleic acid, the stability in serum is increased, and off-target effects are reduced. The effect may be reduced (Elmen, J. et al., (2005) Nucleic Aci ds Research 33(1):439-447;Mook, O R. et al .,(2007)Mol Canc Ther6(3):833-843;Grunwe ller, A. et al., (2003) Nucleic Acids Resear (ch 31(12):3185-3193). dsRNA agents containing loc nucleic acids, AGT Method for producing antisense polynucleotides and / or AGT sense polynucleotides. The method is commonly practiced in the art, and using such a method, a particular modification A of the present invention is GT dsRNA agents can be produced. The AGT dsRNA compound of the present invention, Sen Certain embodiments of polynucleotides and / or antisense polynucleotides are few. It contains at least one modified nucleotide, and at least one of those modified nucleotides is :2' -O-methylnucleotide, 2'-fluoronucleotide, 2'-deoxynucleotide , 2',3'-seconucleotide mimic, locked nucleotide, 2'-F-ara Binonucleotides, 2'-methoxyethyl nucleotides, 2'-amino modified nucleotides , 2'-alkyl-modified nucleotides, morpholino nucleotides and 3'-Ome nucleos Cholesterol, nucleotides containing a 5'-phosphorothioate group, or cholesterol derivatives Alternatively, terminal nucleotides linked to the bisdecylamide group of dodecanoate, 2'-aminobutyric acid Contains ornamental nucleotides, phosphoramidates, or non-natural base-containing nucleotides. In one embodiment, the AGT dsRNA compound is an antisense strand (G in this specification). It contains an E-vinylphosphonate nucleotide at the 5' end of the id chain (also called the id chain).

[0115] In certain embodiments of the present invention, at least one modified nucleotide is AGT d sRNA compounds, the 3' and 5' ends of sense polynucleotides, and / or antisense It is contained at the 3' end of the polynucleotide, and at least one of its modified nucleotides is removed Base nucleotide, ribitol, inverted nucleotide, inverted debased nucleotide, inverted 2' -Contains OMe nucleotides and inverted 2'-deoxynucleotides. Oligonucleoti By including a debased or inverted debased nucleotide at the end of the nucleotide, stability can be increased. It is known to those skilled in the art that this can be done (Czauderna et al. Struc). tural variations and stabilizing modifica ations of synthetic siRNAs in mammalian cells.Nucleic Acids Res.2003;31(11):2705 -2716.doi:10.1093 / nar / gkg393).

[0116] In a specific embodiment of the present invention, an AGT dsRNA compound and an antisense polymer The creotide comprises at least one modified nucleotide, and the at least one modified nucleotide Rheotide is an unlocked nucleic acid (UNA) nucleotide and / or glycol nucleic acid (GNA) ) Contains nucleotides. UNA and GNA have off-target profiles of siRNA compounds. It is known to those skilled in the art that this is a heat-unstable chemical modification that can significantly improve the file. Janas,et al.,Selection of GalNAc-conjuga ted siRNAs with limited off-target-drive n rat hepatotoxicity.Nat Commun 2018;9(1 ):723.doi:10.1038 / s41467-018-02989-4;Lau rsen et al.,Utilization of unlocked nucl eic acid(UNA) to enhance siRNA performance e in vitro and in vivo.Mol BioSyst.2010; 6:862-70).

[0117] The present invention relates to AGT dsRNA agents, AGT antisense polynucleotides, and / or A Another modification that may be present in RNA of a particular embodiment of GT sense polynucleotide is AGT dsRNA agents, AGT antisense polynucleotides and / or AGT sense polynucleotides One or more chemically bound RNAs that enhance one or more characteristics of each rheotide Includes ligands, sites, or conjugates. Non-limiting examples of features that can be enhanced are: : AGT dsRNA agents, AGT antisense polynucleotides and / or AGT sense Polynucleotide activity, cell distribution, delivery of AGT dsRNA agents, AGT dsRNA agents The pharmacokinetic properties and intracellular uptake of AGT dsRNA agents; one aspect of the present invention. In one embodiment of the present invention, the AGT dsRNA agent is, in a particular embodiment of the present invention, The lance chain contains one or more targeted or linking groups conjugated to it. A restrictive example is a compound containing N-acetyl-galactosamine (GalNAc). "Targeting agent," "linking agent," "targeting compound," and "label" In this specification, the term "targeted ligand" may be used without distinction. In specific embodiments of the present invention Furthermore, AGT dsRNA agents have a targeted compound conjugated to the 5' end of the sense strand. Contains a substance. In certain embodiments of the present invention, the AGT dsRNA agent is the 3' of the sense strand. It contains a targeting compound conjugated at the terminal. In some embodiments of the present invention, A GT dsRNA agents contain a GalNAc-containing targeting group. In certain embodiments of the present invention, AGT dsRNA agents have condyloma at one or both of the 3' and 5' ends of the sense strand. It does not contain regulated targeted compounds. In certain embodiments of the present invention, AGT d The sRNA agent is conjugated to one or both of the 5' and 3' ends of the sense strand. Does not contain galNAc-containing targeted compounds.

[0118] Additional targeting and linking agents are well known in the art, for example, the specifics of this invention In the embodiments, useful targeting and coupling agents include, but are not limited to, cholesterol. Lipid regions such as the sacral region (Letsinger et al., Proc. Natl. A cid.Sci.USA,1989,86:6553-6556), cholic acid (Mano haran et al.,Biorg.Med.Chem.Let.,1994,4: 1053-1060), thioates such as beryl-S-tritylthiol (Manoharan et al., Ann. NY Acad. Sci., 1992 ,660:306-309;Manoharan et al.,Biorg.Med. Chem. Let., 1993, 3:2765-2770), thiocholesterol (Ob Erhauser et al., Nucl. Acids Res., 1992, 20: 533-538), aliphatic chains such as dodecanediol or undecyl residues (Saison -Behmoaras et al.,EMBO J,1991,10:1111-11 18;Kabanov et al.,FEBS Lett.,1990,259:32 7-330;Svinarchuk et al.,Biochimie,1993,7 5:49-54), phospholipids, e.g., 2-hexadecyl-rac-glycerol or triglycerides Ethylammonium 1,2-di-O-hexadecyl-rac-glycerol-3-phospho Tetrahedron Lett.,1 995,36:3651-3654;Shea et al.,Nucl.Acids Res., 1990, 18:3777-3783), polyamine or polyethylene glyco chain (Manoharan et al., Nucleosides & Nucle (otides, 1995, 14:969-973) or adamantane acetate (Manoha ran et al.,Tetrahedron Lett.,1995,36:365 1-3654), palmitoyl portion (Mishra et al., Biochim.B iophys.Acta,1995,1264:229-237) or Octadecylami The hexylamino-carbonyloxycholesterol site (Crooke et al., J. Pharmacol. Exp. Ther., 1996, 277: 923- 937) is one example.

[0119] AGT dsRNA agents, AGT antisense polynucleotides, and / or AGT sen Specific embodiments of compositions containing sporinucreoti include the distribution and targeting of AGT dsRNA agents. The present invention may contain ligands that alter properties such as chemical properties. In some embodiments of the substance, for example, a ligand is compared to a species in which such a ligand does not exist. In comparison, the selected target (e.g., molecule, cell or cell type, compartment, e.g., To increase affinity for cells or organ compartments, tissues, organs, or body regions. Ligands useful in the compositions and / or methods of the present invention are proteins (e.g., hi Serum albumin (HSA), low-density lipoprotein (LDL) or globulin, carbohydrates Compounds (e.g., dextran, amylopectin, chitin, chitosan, inulin, cyclodextrin) The ligand may be a natural substance such as chistrin or hyaluronic acid, or a lipid. Alternatively, it may be a synthetic molecule, such as a synthetic polymer like a synthetic polyamino acid or polyamine. Examples of polyamino acids include polylysine (PLL), poly-L-aspartic acid, and poly-L-glycerides. Lutamic acid, styrene-maleic anhydride copolymer, poly(L-lactide-co-glyco (Hydroxylic acid) copolymer, divinyl ether-maleic anhydride copolymer, N-(2-hydro Xypropyl methacrylamide copolymer (HMPA), polyethylene glycol (P EG), polyvinyl alcohol (PVA), polyurethane, poly(2-ethylacrylic acid) ), N-isopropylacrylamide polymer, or polyphosphazene. Examples of ions include: polyethyleneimine, polylysine (PLL), spermine, spermidyl Polyamines, pseudopeptides, peptidomimetic polyamines, dendritic polyamines Amines, arginine, amidine, protamine, cationic lipids, cationic porphyrins Examples include quaternary salts of polyamines or α-helix peptides.

[0120] The ligands included in the compositions and / or methods of the present invention may include a targeting group, and their non-controllable Limited examples include cell or tissue targeting agents, such as lectins, glycoproteins, lipids, or tacin. It is an antibody that binds to specific cell types, such as proteins, including kidney cells or liver cells. The targeting groups are thyroid-stimulating hormone, melanogen, lectin, glycoprotein, and surfactant. Protein A, mucin carbohydrates, polyhydric lactose, polyhydric galactose, N-acetyl-galactose Lactosamine, N-acetyl-glucosamine, polyvalent mannose, polyvalent fucose, glycosyl Polyamino acids, polyvalent galactose, transferrin, bisphosphonate, polyglutamic acid Minic acid, polyaspartic acid, lipids, cholesterol, steroids, bile acids, folic acid, vitamins Vitamin B12, Vitamin A, Biotin, or RGD peptide or RGD peptide mimeti It could be a problem.

[0121] Other examples of ligands include dyes, intercalators (e.g., acridine), and crosslinks. Agents (e.g., psoralen, mitomycin C), porphyrins (TPPC4, texafyl) Sapphyrin, polycyclic aromatic hydrocarbons (e.g., phenazine) Examples of lipophilic molecules include dihydrophenazine, artificial endonucleases (e.g., EDTA), and lipophilic molecules. For example, cholesterol, cholic acid, adamantane acetate, 1-pyrene butyric acid, dihydrotestosterone. Steron, 1,3-bis-O(hexadecyl)glycerin, geranyloxyhexyl, Xadecylglycerol, borneol, menthol, 1,3-propanediol, hep Tadecyl, palmitic acid, myristic acid, O3-(oleoyl)lithocholic acid, O3-( Oleoylcholic acid, dimethoxytrityl or phenoxazine and peptide conjugate (e.g., Antennapedia peptide, Tat peptide), alkylating agent, phosphoric acid, Amino, mercapto, PEG (e.g., PEG-40K), MPEG, [MPEG]2, Polyamino, alkyl, substituted alkyl, radiolabeled markers, enzymes, haptens (e.g., Biotin), transport / absorption enhancers (e.g., aspirin, vitamin E, folic acid), synthesis Ribonucleases (e.g., imidazole, bisimidazole, histamine, imidazo) Lucluster, acridine-imidazole conjugate, Eu of tetraaza macrocyclic molecule 3+ Examples include complexes, dinitrophenyl, HRP, or AP.

[0122] The ligands included in the composition and / or method of the present invention are glycoproteins or peptides, etc. A protein, for example, a molecule that has a specific affinity for a coligand, or an antibody, for example These are antibodies that bind to specific cell types such as cancer cells, endothelial cells, cardiomyocytes, or osteocytes. It is possible. In embodiments of the compositions and / or methods of the present invention, useful ligands are hormones or This can be a hormone receptor. Useful in embodiments of the composition and / or method of the present invention Ligands include lipids, lectins, carbohydrates, vitamins, coenzymes, polyhydric lactose, and polyhydric galacids. Tose, N-acetyl-galactosamine, N-acetyl-glucosamine polyvalent mannose or It can be polyvalent fucose. Useful in embodiments of the composition and / or method of the present invention Gund can destroy cells by, for example, disrupting the cytoskeleton (e.g., microtubules, microtubules). By disrupting the clofilaments and / or intermediate filaments, the cells are introduced into the cell. It may be a substance that increases the uptake of AGT dsRNA agents. Such an unrestricted agonist Examples include: Taxone, vincristine, vinblastine, cytochalasin, nocodazole, di Jaspranquinolide, Latrantrin A, Phalloidin, Swinhold A, Indanosine It may also be myoservin.

[0123] In some embodiments, the ligand bound to the AGT dsRNA agent of the present invention is a drug It plays a role as a kinetic (PK) modifier. In the composition and method of the present invention, Examples of PK modifiers include, but are not limited to, lipophilic agonists, bile acids, and steroids. Phospholipid analogs, peptides, protein binders, PEG, vitamins, cholesterol Fatty acids, cholic acid, lithocholic acid, dialkylglycerides, diacylglycerides, phosphorus Lipids, sphingolipids, naproxen, ibuprofen, vitamin E, biotin, and blood Examples include aptamers that bind to sterile proteins. They contain many phosphorothioate bonds. The oligonucleotides it possesses are also known to bind to serum proteins, therefore Therefore, a short oligonucleotide containing multiple phosphorothioate bonds in its main chain, for example, about Oligonucleotides with 5, 10, 15, or 20 bases are also compositions of the present invention. It may be used as a ligand in and / or by method.

[0124] AGT dsRNA formulation In some embodiments of the present invention, the AGT dsRNA agent is present in the composition. The composition comprises one or more AGT dsRNA agents, and optionally one or more other agents. This may include a carrier, delivery agent, targeting agent, detectable label, etc., which may be permitted. Non-limiting examples of targeting agents that may be useful according to certain embodiments are given in the present invention AGT d An agent that directs sRNA agents to the cells to be treated and / or into the cells to be treated. The selection of a targeting agent varies depending on the nature of the AGT-related disease or condition, and the target cell type. In non-limiting examples, in some embodiments of the present invention, the AGT dsRNA agent is used in hepatocytes. It is desirable to target the cells and / or to target them within hepatocytes. In some embodiments of the present invention The therapeutic agent contains N-acetylgalactosamine (GalNAc) without additional linking elements. Please understand that the AGT dsRNA agent is included along with the delivery agent alone. In some aspects of the present invention, the AGT dsRNA agent is a delivery compound comprising GalNAc A composition containing a carrier that can be attached to an object and is acceptable as a pharmaceutical agent. It is possible to attach a detectable label or targeting agent to the AGT dsRNA agent, etc. It can be administered to cells or subjects in the presence of [the substance].

[0125] The AGT dsRNA agent of the present invention comprises one or more types of delivery agents, targeting agents, labeling agents, etc. They are administered together and / or linked to one or more delivery agents, targeting agents, labeling agents, etc. In this case, a person skilled in the art can select and use an appropriate agent for use in the method of the present invention. It will be understood that the labeling agent is located in cells and tissues, and the position of the AGT dsRNA agent. To determine, the present invention can be used in a particular manner and administered in the manner of the present invention. Used to identify the location of cells, tissues, or organs in therapeutic compositions containing AGT dsRNA agents. Means for attaching and using enzyme labels, dyes, radiolabels, etc., may be used in this art. It is well known. In some embodiments of the compositions and methods of the present invention, the labeling agent is A One or both of the sense and antisense polynucleotides contained in GT dsRNA agents Please understand that it is linked to.

[0126] Delivery of AGT dsRNA agents and AGT antisense polynucleotide agents A particular embodiment of the method of the present invention involves the delivery of an AGT dsRNA agent into a cell. As used in this specification, the term "delivery" refers to promoting intracellular uptake or absorption. This means that it affects intracellular uptake or absorption. AGT dsRNA The absorption or uptake of the agent occurs through independent diffusive or active cellular processes, This occurs by using a delivery agent, targeting agent, etc. that can bind to the AGT dsRNA agent of the present invention. A suitable delivery method for use in the method of the present invention is, but is not limited to, AGT ds RNA agents are injected into tissue sites or administered systemically; this is called in vivo delivery. Examples include: In some embodiments of the present invention, the AGT dsRNA agent is linked to the delivery agent. It will be done.

[0127] It can be used for the delivery of AGT dsRNA agents to cells, tissues, and / or targets. Non-restrictive examples of methods include: AGT dsRNA-GalNAc conjugate, SA Examples include MiRNA technology, LNP-based delivery methods, and naked RNA delivery. Various diseases And the symptoms, though not limited to, include liver disease, acute intermittent porphyria (AIP), hemophilia, and lung disease. These and other delivery methods are used to deliver RNAi therapeutic drugs in the treatment of fibrosis and other conditions. It is well used in the technical field. Details of various delivery methods are available from Nikam, RR & K. .R.Gore(2018)Nucleic Acid Ther,28(4),209 -224 Aug 2018;Springer AD&S.F.Dowdy(20 18) Nucleic Acid Ther.Jun1;28(3):109-118; Lee,K.et al.,(2018)Arch Pharm Res,41(9), 867-874; and Nair, JKet al., (2014) J.Am.Che It is described in publications such as m.Soc.136:16958-16961, and its contents This is incorporated herein by reference.

[0128] Some embodiments of the present invention involve the application of the AGT dsRNA agent of the present invention to cells, tissues and / or other cells. This involves the use of lipid nanoparticles (LNPs) for delivery to elephants. The LNPs are AGT dsRN It is typically used for the in vivo delivery of AGT dsRNA agents such as A therapy agents. LNP or other One advantage of using a delivery agent is that when delivered to the target using LNP or other delivery agents... In addition, the stability of the AGT RNA agent is enhanced. In some embodiments of the present invention The LNP is a cationic molecule loaded with one or more AGT RNAi molecules of the present invention. Contains LNP. LNPs containing AGT RNAi molecules are administered to the subject, and the LNP and its constituents are administered. The combined AGT RNAi molecules are taken up by cells via endocytosis. Its presence triggers the release of RNAi trigger molecules, which in turn mediates RNAi. ru.

[0129] To deliver the AGT dsRNA agent of the present invention to cells, tissues and / or targets, the present invention Another non-limiting example of a delivery agent that may be used in the embodiments is the AGT dsRNA agent of the present invention. It is linked to and delivers the AGT dsRNA agent to cells, tissues, and / or targets, Ga It is an activator containing lNAc. Used in specific embodiments of the methods and compositions of the present invention. Examples of other specific GalNAc-containing delivery agents that may exist are found in PCT application International Publication No. 202019. This is disclosed in pamphlet No. 1183A1. It involves delivering AGT dsRNA agents to cells. Non-GalNAc targeting ligands that can be used in the compositions and methods of the present invention A restrictive example is a targeted ligand cluster. The targeted ligand proposed herein An example of a Gand cluster is: GalNAc liggan with a phosphate diester bond (GLO). It is a GalNAc ligand having a phosphate and phosphorothioate bond (GLS). In this specification, the term "Xn" refers to a compound containing GalNAC that is bonded to the compound. GLS-1, GLS-2, GLS-3, GLS-4, GLS-5, GLS-6, GLS- 7, GLS-8, GLS-9, GLS-10, GLS-11, GLS-12, GLS-1 3, GLS-14, GLS-15, GLS-16, GLO-1, GLO-2, GLO-3 , GLO-4, GLO-5, GLO-6, GLO-7, GLO-8, GLO-9, GLO -10, GLO-11, GLO-12, GLO-13, GLO-14, GLO-15 and It can be used to mean that it is one of the GLO-16s, each The structure is shown below. In the following figure, the GalNAc-targeting ligand and RN of the present invention are shown. The linkage position of the Ai agent is located at the rightmost position of each targeting ligand. And the dsRNA molecules are GLS-1, GLS-2, GLS-3, GLS-4, GLS-5 , GLS-6, GLS-7, GLS-8, GLS-9, GLS-10, GLS-11, G LS-12, GLS-13, GLS-14, GLS-15, GLS-16, GLO-1, GLO-2, GLO-3, GLO-4, GLO-5, GLO-6, GLO-7, GLO- 8, GLO-9, GLO-10, GLO-11, GLO-12, GLO-13, GLO- 14. It should be understood that it can be connected to GLO-15 and GLO-16. The structures of 1-GLO-16 and GLS-1-GLS-16 are shown below. [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka]

[0130] In some embodiments of the present invention, in vivo delivery is described in its entirety by reference herein. Incorporated in U.S. Patent No. 5,032,401 and U.S. Patent No. 5,607,6 The delivery system described in Specification No. 77, U.S. Patent Application Publication No. 2005 / 0281781 Delivery may also be via a β-glucan delivery system such as TEM. AGT RNAi agents are Using methods known in the art, such as electroporation and lipofection, cells are processed in vitro. It can also be introduced into AGT dsRNA. In a specific embodiment of the method of the present invention, AGT dsRNA These RNAs are delivered without targeting agents. These RNAs are delivered as "naked" RNA molecules. It is possible. As a non-limiting example, the AGT dsRNA of the present invention can be used as a targeting agent (e.g., (Not a GalNAc-targeting compound, but administered to the target in a pharmaceutical composition containing an RNAi agent) This allows for the treatment of AGT-related diseases or conditions in patients with conditions such as hypertension.

[0131] In addition to the specific delivery methods described herein, other RNAi delivery methods may be used as described herein. Embodiments of AGT RNAi agents and therapeutic methods, not limited to those described herein, Please understand that this may be used in conjunction with other technologies used in this field.

[0132] The AGT dsRNA agent of the present invention is an AGT polypeptide in cells and / or subjects. The substance can be administered to the subject in an amount and manner effective in reducing its level and activity. In some embodiments, one or more AGT dsRNA agents are administered to the target. AGT expression and activity are associated with diseases or conditions that are treated. In some embodiments, The method of the present invention is intended to alleviate diseases or conditions associated with AGT expression in a subject. To administer one or more AGT dsRNA agents to patients requiring such treatment Includes: Administration of the AGT dsRNA agent or AGT antisense polynucleotide agent of the present invention. AGT expression and / or in one or more cells in vitro, in vitro, and in vivo. This can reduce activity.

[0133] In some embodiments of the present invention, the level of AGT polypeptide in cells, however And that activity is finely detailed by AGT dsRNA agents or AGT antisense polynucleotide agents. It is reduced by delivery (e.g., introduction) into the cell. Using targeted agents and methods, Specific cell types, cell subtypes, organs, or spatial regions within a subject and / or cells within a cell Promote delivery of AGT dsRNA agents or AGT antisense polynucleotide agents to the lower region. It can be advanced. The AGT dsRNA agent, in a specific method of the present invention, alone, Alternatively, it may be administered in combination with one or more additional AGT dsRNA agents. (Some embodiments) In this case, two, three, four, or more independently selected AGT dsRNA agents are used against It is administered to elephants. In certain embodiments of the present invention, the AGT dsRNA agent is AGT-related In conjunction with one or more additional therapies for the treatment of related diseases or conditions, AGT-related diseases or It is administered to the subject to treat the disease. Unrestricted examples of additional treatment regimens are given in this case. Administration of one or more AGT antisense polynucleotides, non-AGT dsRNA This involves the administration of therapeutic drugs and behavioral modifications. Additional treatment regimens are available at the following time points: AG of the present invention. The procedure is performed at one or more of the following time points: before, simultaneously with, and after administration of the T dsRNA agent. It is possible. As used herein, "simultaneously" means within 5 minutes of zero, or within 10 minutes of zero. This means within 30 minutes of zero, within 45 minutes of zero, and within 60 minutes of zero. Therefore, the "zero point in time" is understood to be the point in time when the AGT dsRNA agent of the present invention is administered to the subject. I want to unravel this. Non-limiting examples of non-AGT dsRNA therapies include: diuretics, angiotensin mutations. ACE inhibitors, angiotensin II receptor antagonists, beta-blockers, blood vessels Dilators, calcium channel blockers, aldosterone antagonists, α-2 agonists Stromal, renin inhibitors, alpha-blockers, peripheral adrenergic agonists, selective D1 receptor moieties Agonist, non-selective α-adrenergic antagonist, synthetic, steroidal antimineralicoid , or additional therapeutic agents such as any combination of the above, and formulations into pharmaceutical combinations. It is a drug used to treat hypertension. Non-restrictive behavioral modifications include: administration plan, counseling and These are exercise therapies. These, and other therapeutic agents and behavioral modifiers are publicly known in the art. It may be used to treat AGT disease or condition in the subject, and AGT disease or condition For therapeutic purposes, it may be administered to the target in combination with one or more AGT dsRNA agents of the present invention. The present invention provides an AG that is administered to cells or subjects to treat AGT-related diseases or conditions. T dsRNA agents act synergistically with one or more other therapeutic agents or active ingredients. This allows for the effectiveness of one or more other therapeutic agents or active ingredients to be enhanced. AGT dsRNA is used in the treatment of AGT-related diseases or conditions. The effectiveness of the agent is enhanced.

[0134] The therapeutic method of the present invention is used before the onset of AGT-related disease or condition, and / or AGT-related disease When a disease or illness exists, for example, in the early, middle, and late stages of the disease or illness, AGT dsRNA agents can be used at any time before or after any of these stages. The method of the present invention is used in cases where treatment of the target AGT-related disease or condition has been unsuccessful. , minimal success and / or no longer success, one or more other A therapeutic agent and / or therapeutic active ingredient for AGT-related diseases or conditions that have been treated in advance. Even those who are already being treated can be treated.

[0135] Vector-encoding dsRNA In a particular embodiment of the present invention, in order to deliver the AGT dsRNA agent to cells, A der can be used. The AGT dsRNA agent transcription unit is DNA or RNA. This may be included in the processor. Such transgene codes for delivering sequences to cells and / or targets. The preparation and use of ionization vectors are well known in the art. For example, at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 hours or more, or at least 1, 2 For 3, 4, 5, 6, 7, 8, 9, 10 weeks or more, one AGT dsRNA A vector that causes transient expression can be used in the method of the present invention. The length is not limited, but is a selected specific vector construct, and target cells and / or Such transgenes can be determined using conventional methods based on factors such as tissue. linear constructs, circular plasmids, or viruses, which may be embedded or unembedded vectors. It can be introduced as a vector. The introduced gene is inherited as an extrachromosomal plasmid. It is also possible to create transgenes so that they can be used (Gassmann, et al., P roc.Natl.Acad.Sci.USA(1995)92:1292).

[0136] One or more single strands of AGT dsRNA agents are transmitted from the promoter on the expression vector. It can be transcribed. For example, if two separate strands are expressed to produce dsRNA. In this, two separate expression vectors are used by means of transfection or infection. It can be introduced into cells simultaneously. In certain embodiments, the individual AGT dsRNA agent of the present invention Each of these strands can be transcribed from a promoter contained on the same expression vector. In a specific embodiment of the present invention, the AGT dsRNA agent is the stem (s Linked by linker polynucleotide sequences to have a tem and loop structure It is expressed as an inverted repeat polynucleotide.

[0137] Unrestrictive examples of RNA expression vectors are DNA plasmids or viral vectors. An expression vector useful in the embodiments of the present invention may be compatible with eukaryotic cells. Biological expression vectors are commonly used in this field and are available from many commercial suppliers. It is possible. Delivery of the AGT dsRNA expression vector can be done, for example, by intravenous or intramuscular administration. By means of administration to target cells that are extracted from the subject and reintroduced into the subject, or for the purpose This may involve systemic delivery by any means that enables the introduction of target cells.

[0138] The viral vector systems that may be included in the embodiments of the method are, but are not limited to: a) Adenovirus vectors; (b) Retrovirus vectors, not limited to, but Viral vectors, Moloney's mouse leukemia virus, etc.; (c) Adeno-associated virus vectors (d) Herpes simplex virus vector; (e) SV40 vector; (f) Polyo (g) Mavirus vector; (h) Papillomavirus vector; (g) Picornavirus vector (i) Poxvirus vectors, for example, orthopoxvirus vectors, for example, Vaccinia virus vectors or canary or poultry poxvirus vectors, etc. Tripoxvirus vector; (j) Helper-dependent or gutless Adenovirus vectors are an example. They are used for the recombinant expression of AGT dsRNA agents. The constructs may be selected to provide constitutive or regulatory / inducible expression, promoters, It may contain regulatory factors such as enhancers. Viral vector systems and promoters - The use of enhancers, etc., is common practice in the art and is described herein. It can also be used in combination with other compositions.

[0139] A particular embodiment of the present invention is a viral delivery method for delivering AGT dsRNA agents to cells. This includes the use of ctor. Many adenovirus-based delivery systems are used, for example, in the lungs, liver, It is commonly used in this art for delivery to the central nervous system, endothelial cells, and muscles. Non-limiting examples of viral vectors that may be used in the method of the present invention include: AAV vector, vaccine Poxviruses such as Senior Virus, modified Ankara virus (MVA), NYVAC or tripoxviruses such as poultry or canary poxvirus.

[0140] A particular embodiment of the present invention involves delivering an AGT dsRNA agent to cells using a vector. The method includes a sustained-release matrix into which the gene delivery vector is embedded. It is not necessary to include, but may include, and may be a pharmacopoeially acceptable vector. Morphologically, vectors for delivering AGT dsRNA are produced by recombinant cells. The pharmaceutical composition of the present invention can produce an AGT dsRNA delivery system. It may contain one or more types of cells.

[0141] Pharmaceutical composition containing AGT dsRNA or ssRNA agent Specific embodiments of the present invention relate to AGT dsRNA agents or AGT antisense polynucleotides. This includes the use of a pharmaceutical composition containing an otidol and a pharmacopoecitable carrier. Pharmaceutical compositions containing dsRNA agents or AGT antisense polynucleotide agents are detailed Used in the method of the present invention to reduce AGT gene expression and AGT activity in cells Such medicine can be used to treat related diseases or conditions. Compositions can be formulated based on the delivery method. Non-restrictive examples of formulations for different delivery methods include: : Compositions formulated for subcutaneous delivery, compositions formulated for systemic administration by parenteral delivery Substances, compositions formulated for intravenous (IV) delivery, compositions formulated for intrathecal delivery, and compositions formulated for direct delivery into the brain. The pharmaceutical compositions of the present invention are To deliver AGT dsRNA agents or AGT antisense polynucleotide agents to cells Using one or more means, for example: topical administration (e.g., via a transdermal patch); intrapulmonary administration. For example, by inhaling or blowing powder or aerosol via a nebulizer. Administration; it can be administered intratracheally, intranasally, epidermally and transdermally, orally or parenterally. Parenteral administration. Administration methods include intravenous, intra-arterial, subcutaneous, intraperitoneal, or intramuscular injection or infusion; for example, implantation. Subcutaneous administration via a device; or intracranial administration, for example, intraparenchymal, intrathecal, or intraventricular. It may be administered using AGT dsRNA agents or AGT antisense polynucleotides. Tydotherapy drugs can also be delivered directly to target tissues, such as directly to the liver or directly to the kidneys. To "deliver AGT dsRNA agents" or "deliver AGT antisense polynucleotide agents" "To deliver" means to deliver an AGT dsRNA agent or an AGT antisense polynucleotide agent. To deliver them, to directly express AGT dsRNA agents in cells, and cells Expressing AGT dsRNA agents from an encoding vector delivered to cells, or in cells Either AGT dsRNA or AGT antisense polynucleotide agent is present. It should be understood that this includes appropriate means of preparation and use of formulations, and delivery of inhibitory RNA. The means of achieving this are well known and commonly used in this field.

[0142] As used herein, "pharmaceutical composition" refers to a pharmacologically effective amount of AGT dsR of the present invention. A NA agent or AGT antisense polynucleotide agent and a carrier that is acceptable as a drug, Includes. The term "pharmaceutically acceptable carrier" is used to administer therapeutic drugs. It means a carrier. Such carriers are not limited to physiological saline, buffered physiological saline, Examples include glucose, water, glycerol, ethanol, and combinations thereof. Specifically, the term excludes cell culture media. Regarding orally administered drugs, the term is permitted as a drug. Acceptable carriers are not limited to, but include excipients that are acceptable as pharmaceuticals, such as inactive dilutes. Examples include disintegrants, disintegrants, binders, lubricants, sweeteners, flavorings, colorings, and preservatives. Appropriate Suitable inert diluents include sodium carbonate and calcium carbonate, sodium phosphate and ri Examples include calcium phosphate and lactose, and suitable corn starch and alginic acid. It is a disintegrant. Examples of binders include starch and gelatin, and a white lubricant is used. If present, it is usually magnesium stearate, stearic acid, or talc. If desired, the tablets may contain glyceryl monostearate to slow down absorption in the gastrointestinal tract. It may be coated with materials such as phosphorus or glyceryl distearate. Actions included in pharmaceutical formulations. The agent is further described below. The term "pharmacologically effective" as used herein refers to the pharmacologically effective agent used in this specification. Terms such as "quantity," "therapeutic effective dose," and "effective dose" refer to the intended pharmacological, therapeutic, or The AGT dsRNA agent or AGT antisense polynucleotide of the present invention that produces a preventive outcome This refers to the amount of leotide. For example, it refers to the measurable parameter associated with the disease or disorder. A prescribed clinical treatment is considered effective if the dose is reduced by at least 10%. The therapeutically effective dose of the drug used to treat the disease or condition is determined by its parameters. - This is an amount that needs to be reduced by at least 10%. For example, AGT dsRNA agent or The therapeutically effective dose of AGT antisense polynucleotide agents is low in AGT polypeptide levels. It can be reduced by 10% even without it. The pharmaceutical compositions are AD00051~AD shown in Table 1. 00122-19-2, AD00163-3, AV01227~AVAV01257, and It may include dsRNAi agents containing double hemispheres such as AV01711. For example, preferred dsRNAi agents include double-stranded AD00158 and AD00163. Examples include AD00159, AD00290, AD00300, or AD00122. In other embodiments, preferred dsRNAi agents include, for example, AD00158- 1, AD00158-2, AD00163-1, AD00159-1, or AD0030 Examples include 0-1. In some other embodiments, such dsRNAi agents include two Heavy chain variants, e.g., double chain AD00158, AD00163, AD00163-3 Variants of AD00159, AD00290, AD00300 or AD00122 It can be listed.

[0143] Effective amount In some embodiments, the method of the present invention reduces AGT gene expression in contact cells. Therefore, an effective amount of AGT dsRNA agent or AGT antisense polynucleotide agent is used. This includes contacting cells. A particular embodiment of the method of the present invention reduces AGT gene expression. Furthermore, in an amount effective to treat AGT-related diseases or conditions in the subject, AGT ds This includes administering RNA agents or AGT antisense polynucleotide agents to the target. Use to reduce T expression and / or for the treatment of AGT-related diseases or conditions. In this case, the "effective dose" is the amount necessary or sufficient to perceive the desired biological effect. For example, AGT dsRNA agents or AGT for treating AGT-related diseases or conditions. The effective dose of antisense polynucleotide agents slows or stops the progression of the disease or condition. (ii) To stop; (ii) To reverse, reduce, or reverse one or more symptoms of a disease or condition. It may be the amount necessary to resolve; in some embodiments of the present invention, the effective amount is related to AGT When administered to a patient requiring treatment for a disease or condition, it may produce a therapeutic response, and the disease or The disease is prevented and / or treated by AGT dsRNA agents or AGT antisense polymerases. This is the amount of nucleotide agent. According to some aspects of the present invention, the effective amount is for AGT-related diseases or When administered in combination with or concurrently with therapeutic measures specific to the disease, it elicits a therapeutic response. AGT dsRNA agents or AGT antiseptics are used to prevent and / or treat diseases or conditions. This is the amount of the nspolynucleotide agent. In some embodiments of the present invention, the AGT of the present invention The biological effect of treating the target with dsRNA agents or AGT antisense polynucleotide agents. The effect may be the alleviation and / or complete elimination of symptoms caused by AGT-related diseases or conditions. In some embodiments of the present invention, the biological effect is, for example, that the subject has AGT-related disease. Or, AGT-related disease or complete absence of symptoms as demonstrated by diagnostic tests showing the absence of symptoms. This is an effective blockage. A non-limiting example of detectable physiological symptoms is the administration of the agent of the present invention. Later, a reduction in lipid accumulation in the target liver is mentioned. AGT-related disease or medical condition Using other means known in the art to evaluate AGT-related diseases or conditions The effects of the agent and / or method of the present invention can be determined in relation to the present invention.

[0144] A to reduce the activity of AGT polypeptides to a level that treats AGT-related diseases or conditions. The effective dose of GT dsRNA agents or AGT antisense polynucleotide agents was determined in clinical trials. The effective dose is determined and established in blinded trials of the test population compared to the control population. In the embodiment, the effective amount is the cells, tissues, and / or cells affected by the disease or condition. This is the amount that produces the desired response, such as a reduction in AGT-related disease or symptoms in the target population. Yes. Therefore, AGT activity can be treated by reducing AGT polypeptide activity. AGT dsRNA agents or AGT antisense polymers for treating related diseases or conditions The effective dose of nucleotide agents, when administered, is equivalent to that of AGT dsRNA agents or AGT antiseptics. The amount that would be present in cells, tissues, and / or subjects that are not administered the nspolynucleotide agent. A lower amount may be an amount that reduces the amount of AGT polypeptide activity in the target. In certain embodiments, the AGT dsRNA agent or AGT antisense polynucleotide of the present invention Present in cells, tissues, and / or subjects that have not been exposed to or administered leotide agents. The level of AGT polypeptide activity and / or AGT gene expression is referred to as the "control" level. In some embodiments of the present invention, the control amount of the subject is the amount of the subject before treatment. In other words, the level in the subject before administration of the AGT agent may be the control level of the subject, AGT polypeptide activity and / or AGT gene activity in subjects after administration of siRNA to elephants. It can be used for comparison with the level of gene expression. In the treatment of AGT-related diseases or conditions. In this case, the desired response is one of the diseases or conditions in the cells, tissues, and / or subject. Multiple symptoms may be reduced or eliminated. This reduction or elimination may be temporary. Or it may be permanent. AGT polypeptide activity, AGT gene expression, symptom evaluation, clinical By using methods to determine the tests, etc., it is possible to monitor the state of AGT-related diseases or conditions. Please understand that this is possible. In some embodiments of the present invention, treatment of AGT-related diseases or conditions The desired response to this is to delay or even prevent the onset of symptoms or conditions.

[0145] The effective amount of a compound that reduces the activity of AGT polypeptide is the amount of AGT-related disease after administration or AGT dsRNA agents or AGT antisenses applied to cells or targets, such as reducing disease symptoms. This can also be determined by evaluating the physiological effects of administering renucleotide drugs. Using assays and / or symptom monitoring in the subject, the AGT ds of the present invention RNA agents or AGT antisense polynucleotide agents (administered in the pharmaceutical compounds of the present invention) It is possible to determine the effectiveness of (and whether there is a response to the treatment) It can be determined. A non-limiting example is one or more blood pressure tests known in the art. There is another non-limiting example: before and after treating a subject with the AGT dsRNA agent of the present invention. , to determine the state of the AGT-related disorder of the subject, one or more blood samples known in the art This is a pressure test. In another non-limiting example, to lower blood pressure levels, 1 known in the art The status of AGT-related disease in the subjects is determined using one or more tests. In this study, the disease included hypertension, and using the AGT dsRNA agent of the present invention, The reduction in blood pressure levels in the subjects before and after treatment is determined.

[0146] Some embodiments of the present invention relate to one or more AGT-related diseases or conditions in the subject. By evaluating and / or monitoring the "physiological characteristics," AGT-related diseases or illnesses can be identified. The dsRNA agent or AGT antisense agent of the present invention administered to a subject to treat the condition Includes a method for determining the efficacy of a renucleotide agent. Physiological characteristics of AGT-related diseases or conditions. Non-limiting examples of indicators include serum AGT levels, mean arterial pressure, and diastolic blood pressure in subjects. Standard methods for determining such physiological characteristics are publicly known in the art and are not limited thereto. However, these include blood tests, imaging studies, and physical examinations.

[0147] AGT dsRNA agents or AGT antisense polynucleotide agents administered to the target The amount is determined by the subject, the determinants of the disease and / or the state of the disease and / or life It should be understood that modifications can be made based on at least some of the physical characteristics. AGT-dsR To increase or decrease the amount of NA agent or AGT antisense polynucleotide agent, for example If an AGT dsRNA agent or an AGT antisense polynucleotide agent is administered within it... By changing the composition, by changing the administration route, the timing of administration can be changed. By doing so, the treatment dose can be changed. AGT dsRNA agent or AGT The effective dose of nitrescent polynucleotides depends on the specific medical condition being treated and the target patient. Age and medical condition, severity of condition, duration of treatment, nature of concurrent treatment (if any), specific administration It varies depending on the route, as well as other factors within the knowledge and professional opinion of a healthy physician. For example, The efficacy is determined by the AGT polypeptide activity and / or AG activity that is effective in treating AGT-related diseases or conditions. This may vary depending on the desired level of T gene expression. Those skilled in the art will know from experience that it is not necessary to experiment more than necessary. Without performing any action, the specific AGT dsRNA agent or AGT anti used in the method of the present invention The effective amount of sense polynucleotide agent can be determined. Teachings provided herein Along with the various AGT dsRNA agents or AGT antisense polynucleotides of the present invention, The agent, and the factors that are important, such as potency, relative bioavailability, patient weight, By selecting from the severity of adverse side effects and preferred administration methods, specific It is possible to devise effective preventive or therapeutic therapies to effectively treat the target. The AGT dsRNA agent or AGT agent used in embodiments of the present invention The effective amount of cysens polynucleotide agent is such that, upon contact with it, it produces the desired bioavailability in cells. It can be a quantity that produces a physical effect.

[0148] AGT gene silencing can be constitutive or performed on any cell expressing AGT. This can be done through genome engineering and determined by an appropriate assay. It will be recognized that this is possible. In some embodiments of the present invention, AGT gene expression By administering the AGT dsRNA agent of the present invention, at least 5%, 6%, and 7% of the population are affected. %, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or It is reduced by 100%. In some embodiments of the present invention, AGT gene expression is the present invention By administering AGT dsRNA agents, the rates increase by 5-10%, 5-25%, and 10-50%. It is reduced by 10-75%, 25-75%, 25-100%, or 50-100%.

[0149] dosage AGT dsRNA agents and AGT antisense polynucleotide agents are used for the AGT gene. It is delivered in a pharmaceutical composition in a dose sufficient to inhibit expression. Specific embodiments of the present invention In this state, the dose of AGT dsRNA agent or AGT antisense polynucleotide agent is , 0.01 to 200.0 mg per kilogram of recipient's body weight per day, generally 1 ~50mg / kg body weight, 5~40mg / kg body weight, 10~30mg / kg body weight, 1~20 The dosage is mg / kg body weight, 1-10 mg / kg body weight, or 4-15 mg / kg body weight / day. For example, a single dose of either an AGT dsRNA agent or an AGT antisense polynucleotide agent is , about 0.01mg / kg, 0.05mg / kg, 0.1mg / kg, 0.2mg / kg, 0.3mg / kg, 0.4mg / kg, 0.5mg / kg, 1mg / kg, 1.1mg / kg, 1.2mg / kg, 1.3mg / kg, 1.4mg / kg, 1.5mg / kg, 1 .6mg / kg、1.7mg / kg、1.8mg / kg、1.9mg / kg、2mg / k g、2.1mg / kg、2.2mg / kg、2.3mg / kg、2.4mg / kg、2. 5mg / kg、2.6mg / kg、2.7mg / kg、2.8mg / kg、2.9mg / kg、3.0mg / kg、3.1mg / kg、3.2mg / kg、3.3mg / kg、3 .4mg / kg、3.5mg / kg、3.6mg / kg、3.7mg / kg、3.8mg / kg、3.9mg / kg、4mg / kg、4.1mg / kg、4.2mg / kg、4. 3mg / kg、4.4mg / kg、4.5mg / kg、4.6mg / kg、4.7mg / kg、4.8mg / kg、4.9mg / kg、5mg / kg、5.1mg / kg、5.2 mg / kg、5.3mg / kg、5.4mg / kg、5.5mg / kg、5.6mg / k g、5.7mg / kg、5.8mg / kg、5.9mg / kg、6mg / kg、6.1m g / kg、6.2mg / kg、6.3mg / kg、6.4mg / kg、6.5mg / kg 、6.6mg / kg、6.7mg / kg、6.8mg / kg、6.9mg / kg、7mg / kg、7.1mg / kg、7.2mg / kg、7.3mg / kg、7.4mg / kg、 7.5mg / kg、7.6mg / kg、7.7mg / kg、7.8mg / kg、7.9m g / kg、8mg / kg、8.1mg / kg、8.2mg / kg、8.3mg / kg、8 .4mg / kg、8.5mg / kg、8.6mg / kg、8.7mg / kg、8.8mg / kg、8.9mg / kg、9mg / kg、9.1mg / kg、9.2mg / kg、9. 3mg / kg、9.4mg / kg、9.5mg / kg、9.6mg / kg、9.7mg / kg、9.8mg / kg、9.9mg / kg、10mg / kg、11mg / kg、12m g / kg, 13mg / kg, 14mg / kg, 15mg / kg, 16mg / kg, 17m g / kg, 18mg / kg, 19mg / kg, 20mg / kg, 21mg / kg, 22m g / kg, 23mg / kg, 24mg / kg, 25mg / kg, 26mg / kg, 27m g / kg, 28mg / kg, 29mg / kg, 30mg / kg, 31mg / kg, 32m g / kg, 33mg / kg, 34mg / kg, 35mg / kg, 36mg / kg, 37m g / kg, 38mg / kg, 39mg / kg, 40mg / kg, 41mg / kg, 42m g / kg, 43mg / kg, 44mg / kg, 45mg / kg, 46mg / kg, 47m It can be administered in doses ranging from g / kg, 48 mg / kg, or 49-50 mg / kg body weight.

[0150] When determining the dosage and delivery timing of the AGT dsRNA agent of the present invention, various Factors that may be considered include the delivered AGT dsRNA agent or AGT antisense polymerase. The absolute amount of creotide drugs depends on simultaneous treatment, number of administrations, and age, physical condition, size, and weight. It varies depending on various factors, such as the individual target parameters. These factors are known to those skilled in the art. This is well known and can be addressed through conventional experiments. In some embodiments Therefore, the maximum dose, that is, the safest dose according to sound medical judgment, may be used.

[0151] In some embodiments, the method of the present invention uses an AGT dsRNA agent or an AGT antiseptic. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 doses of nspolynucleotide preparation or more This may include administering the above doses to the target population. In some cases, the dose of the pharmaceutical compound (e.g., (containing AGT dsRNA agents or AGT antisense polynucleotide agents) It can be administered to the target at least daily, every other day, weekly, every other week, monthly, etc. The dosage is daily One or more times, for example, two, three, four, five or more times within a 24-hour period. It can be administered in multiple doses. The pharmaceutical composition of the present invention can be administered once a day; or AG T dsRNA agents or AGT antisense polynucleotide agents should be used throughout the day. It can be administered in two, three or more partial doses at short intervals, or by continuous infusion or sustained release. It can be administered by delivery by a drug. In some embodiments of the present invention, The medicinal composition of the present invention may be taken once or more times a day, once or more times a week, once or more times a month, or It is administered to the target individual once or multiple times a year.

[0152] In a particular embodiment, the method of the present invention involves using a pharmaceutical compound alone, or one or more other A In combination with a GT dsRNA agent or an AGT antisense polynucleotide agent, and / Or other drug therapies or treatments administered to subjects suffering from AGT-related diseases or conditions. This includes administering the drug in combination with an activity or administration plan. The pharmaceutical compound may be in the form of a pharmaceutical composition. It can be administered in a sterile state. The pharmaceutical composition used in the method of the present invention is sterile and can be administered to the target. AGT powder in a suitable weight or volume to a level sufficient to produce the desired reaction AGT dsRNA agents or AGT antisense polynucleotides that reduce the activity of lipeptides. It may contain an amount of otide. It is administered to the subject to reduce AGT protein activity. A pharmaceutical composition containing an AGT dsRNA agent or an AGT antisense polynucleotide agent The dosage is selected according to various parameters, particularly the administration method used and the patient's condition. Obtain. Other factors include the required duration of treatment. Target reaction with the initial dose. If the response is insufficient, a higher dose may be administered, as long as the patient can tolerate it. The dose can be increased effectively (or via a different, more localized delivery route).

[0153] treatment Diseases and disorders that may benefit from reduced AGT gene expression as used herein. When used to mean a medical condition, the terms "prevention" or "preventing" are, The target is, for example, the renin-angiotensin-aldosterone system (RAAS) in cases of hypertension. A disease, disorder, or medical condition that is caused by or associated with the activation of ) This means that the likelihood of developing related symptoms is low. In such situations, hypertension The likelihood of developing the disease decreases; for example, an individual has one or more risk factors for hypertension. , either not developing hypertension, or only developing mild hypertension, or disease , not developing a disability or medical condition, or symptoms associated with such disease, disability, or medical condition The incidence of the condition is higher compared to the same risk factor population that has not received the treatment described herein. Reduced (for example, at least the scale of disease or condition in a clinical setting) (10% reduction), or delayed onset of symptoms (e.g., by days, weeks, months, or years). In such cases, prevention is considered effective.

[0154] Based on the average of sitting blood pressure readings accurately measured during two or more visits, normal blood The target pressure range is systolic blood pressure of approximately 90-119 mmHg (approximately 12-15.9 kPa (kN / m²)). 2 ) ) and diastolic blood pressure approximately 60-79 mmHg (approximately 8.0-10.5 kPa (kN / m²)2 ) has. The subjects with prehypertension have a systolic blood pressure of about 120 - 139 mmHg (about 16.1 - 18 .5 kPa (kN / m .5kPa(kN / m 2 )) and a diastolic blood pressure of about 60 - 79 mmHg (about 8.0 - 10.5 kPa(kN / m 2 )); the subjects with hypertension (e.g., stage I hypertension) have a systolic blood pressure of about 140 - 159 mmHg (about 18.7 - 21.2 kPa (kN / m )) and 2 ) and a diastolic blood pressure of about 90 - 99 mmHg (about 12.0 - 13.2 kPa (kN / m 2 )); the subjects with hypertension (e.g., stage II hypertension) have a systolic blood pressure of about ≧160 mmHg (about ≧21.3 kPa (kN / m≧)) and a diastolic blood pressure of about ≧100 mmHg (about ≧13. 3 kPa (kN / m )) has. 2 ) has.

[0155] In certain embodiments, the angiotensinogen-related disease is essential hypertension. "Essential hypertension" is the result of environmental or genetic factors (e.g., a result without a clear, underlying medical cause ). Result).

[0156] In certain embodiments, the angiotensinogen-related disease is secondary hypertension. "Secondary hypertension" has an identifiable underlying condition and has multiple etiologies such as renal, vascular, and endocrine causes , e.g., renal parenchymal diseases (e.g., polycystic kidney disease, glomerular or interstitial diseases), renal vascular diseases( e.g., renal artery stenosis, fibromuscular dysplasia), endocrine disorders (e.g., adrenocortical steroid or mineralocorticoid excess, pheochromocytoma, hyperthyroidism or hypothyroidism, growth hormone excess, hyperparathyroidism), coarctation of the aorta, or use of oral contraceptives, etc. ). ).

[0157] In certain embodiments, angiotensinogen-related diseases include malignant hypertension and accelerated hypertension. This refers to hypertensive emergencies such as high blood pressure. "Accelerated hypertension" is defined as the condition of one or more terminal organs. Severe blood pressure elevation with direct damage (i.e., systolic blood pressure of 180 mmHg or higher or This means a diastolic blood pressure of 110 mmHg. Blood pressure is immediately important to prevent further organ damage. It must be lowered. "Malignant hypertension" is defined as direct hypertension to one or more terminal organs. Severe hypertension (i.e., systolic blood pressure of 180 mmHg or higher) accompanied by damage and papilledema. This means a diastolic blood pressure of 110 mmHg. Blood pressure is important to prevent further organ damage. It must be lowered to a seat. Nerve terminals caused by uncontrolled blood pressure. Damage to the brain includes hypertensive encephalopathy, cerebrovascular disease / cerebral infarction, subarachnoid hemorrhage and / or intracranial hemorrhage. Bleeding is a possible cause. Damage to the cardiovascular terminal organs includes myocardial ischemia / infarction and acute left ventricular dysfunction. Other organ systems may also show acute renal failure / Uncontrolled It can be affected by high blood pressure.

[0158] In certain embodiments, angiotensinogen-related disease is acute hypertension. "Hypertensive hypertension" is a severe condition characterized by elevated blood pressure that does not directly damage one or more organs. This means an increase (i.e., a systolic blood pressure of 180 mmHg or higher, or a diastolic blood pressure of 110 mmHg). Blood pressure can be safely lowered within a few hours.

[0159] In certain embodiments, angiotensinogen-related disorders include pregnancy-related hypertension, for example. Chronic hypertension during pregnancy, gestational hypertension, pre-eclampsia, eclampsia, and eclampsia aggravated by chronic hypertension. Pre-eclampsia, HELLP syndrome, and pregnancy-induced hypertension (transient pregnancy-induced hypertension, seen in late pregnancy) This is chronic hypertension, also known as gestational hypertension (PIH). The target group for this condition is those whose blood pressure exceeds 140 / 90 mmHg before pregnancy or 20 weeks before pregnancy. It is an elephant. "Pregnancy-induced hypertension" or "pregnancy-induced hypertension" is a condition that, without other characteristics of pre-eclampsia, This refers to hypertension that develops in late pregnancy (after 20 weeks of gestation) and returns to normal after delivery. Mild preeclampsia occurs in women with normal blood pressure at least 6 hours before 20 weeks of pregnancy. However, there is no evidence of terminal organ damage, and two epidemiological conditions with hypertension (blood pressure ≥ 140 / 90 mmHg) It is defined as a sword. In subjects with pre-existing essential hypertension, pre-eclampsia is characterized by systolic contractions. Diagnosis is made when systolic blood pressure rises by 30 mmHg or diastolic blood pressure rises by 15 mmHg. "Severe pre-eclampsia" is defined as the following signs or symptoms of pre-eclampsia: symptoms appear after at least 6 hours. Two episodes in which systolic blood pressure was 160 mmHg or higher, or diastolic blood pressure was 110 mmHg or higher; Proteinuria exceeding 5g collected over a 24-hour period, or collected at least 4 hours apart. Two randomly selected urine samples show a result of 3+ or higher; pulmonary edema or cyanosis; excessive urine volume. Small amount (less than 400 mL in 24 hours); persistent headache, epigastric pain, and / or liver dysfunction; blood It is defined as the presence of one of the following: thrombocytopenia, oligohydramnios, fetal growth restriction, or placental abruption. Eclampsia is defined as a seizure in a woman with pre-eclampsia that cannot be caused by any other factor. It is called "HELLP syndrome" (edema-proteinuria-hypertension, also known as type B pre-eclampsia). (This refers to hemolysis, elevated liver enzyme levels, and decreased platelet levels in pregnant subjects.) It means a small amount.

[0160] In certain embodiments, angiotensinogen-related disease is treatment-resistant hypertension. "Treatment-resistant hypertension" is a condition in which one of the antihypertensive drugs is thiazide diuretics. Despite the simultaneous use of three different classes, the value exceeded the target value mentioned above (for example, 140). This refers to blood pressure that remains at 90 mmHg. Blood pressure is controlled with four or more medications. Those who underwent this procedure are also considered to have treatment-resistant hypertension.

[0161] A decrease in AGT polypeptide levels and / or activity is beneficial in the treatment of a disease or condition. The efficacy of the present invention lies in treating AGT-related diseases and conditions using the method and AGT dsRNA agent of the present invention. This treatment can inhibit AGT expression. The AGT dsRNA agent or AG of the present invention T antisense polynucleotide agents and diseases and illnesses that can be treated by the method of the present invention Examples of the condition include, but are not limited to, hypertensive diseases, hypertension, borderline hypertension, and essential hypertension. Hypertension, secondary hypertension, isolated systolic or diastolic hypertension, pregnancy-related hypertension, diabetic hypertension Blood pressure, treatment-resistant hypertension, refractory hypertension, paroxysmal hypertension, renovascular hypertension, Goldb Rat hypertension, intraocular pressure, glaucoma, pulmonary hypertension, portal hypertension, systemic venous hypertension, systolic Hypertension, unstable hypertension; hypertensive heart disease, hypertensive nephropathy, atherosclerosis, Cardiac sclerosis, vascular disease, diabetic nephropathy, diabetic retinopathy, chronic heart failure, cardiomyopathy, diabetic heart Myopathy, glomerulosclerosis, aortic stenosis, aortic aneurysm, ventricular fibrosis, heart failure, myocardial infarction, anguina , stroke, kidney disease, renal failure, systemic sclerosis, intrauterine growth restriction (IUGR), and fetal development Delays are one example. Such diseases and conditions are referred to as "AGT-related diseases and conditions" in this specification. These may be called "diseases and conditions caused and / or regulated by AGT."

[0162] In certain embodiments of the present invention, the AGT dsRNA agent or AGT antisense agent of the present invention Polynucleotide preparations are administered once or multiple times before or after the diagnosis of AGT-related disease or condition. It can be administered to. In some embodiments of the present invention, the subject is AGT-related disease or disease There is a risk of developing or contracting an AGT-related disease or condition. In some subjects, the risk of developing AGT-related disease or condition is higher compared to the control group. This refers to individuals who are more likely to develop the disease or illness. In some embodiments of the present invention, The risk level is statistically significant compared to the control level of risk. For example, the subjects should be those related to AGT, rather than control subjects without pre-existing diseases or genetic abnormalities. The subject has a pre-existing disease and / or genetic abnormality that makes them more susceptible to developing related diseases or conditions. Subjects who are or will be subject to; family history of AGT-related diseases or conditions and / or Subjects with a personal history; and subjects who have previously received treatment for AGT-related diseases or conditions; This includes pre-existing conditions and / or sequelae that make the subject more susceptible to AGT-related diseases or conditions. If a genetic abnormality is present, it increases the likelihood of developing AGT-related diseases or conditions. It should be understood that this may be related to a previously determined disease or genetic abnormality. .

[0163] AGT dsRNA agents or AGT antisense polynucleotide agents are used for each target disease. Please understand that it may be administered to the subject based on their condition. For example, the health care provided to the subject By this method, the AGT level measured in the sample taken from the subject can be evaluated, and this Administer the AGT dsRNA agent or AGT antisense polynucleotide agent of the invention. This can help determine whether it is desirable to reduce the target AGT level. In limited cases, biological samples such as blood or serum samples are taken from the subject, and the subject's AG The T level can be determined in the sample. AGT dsRNA agent or AGT antisense agent The subject was administered a nucleotide agent, and after administration, a blood or serum sample was collected from the subject. Use the sample to determine the AGT level and compare the result with the pre-administration (previous) sample of the subject. The decrease in the subject's AGT level in subsequent samples compared to the pre-administration level indicates the subject's AG In reducing T levels, administered AGT dsRNA agents or AGT antisense polymerase This demonstrates the efficacy of nucleotide agents. In a non-restrictive example, even if the subject is not described herein, Even if not diagnosed with an AGT-related disorder such as a blood pressure disorder, blood pressure is a physiological characteristic of related disorders. It may be considered a sign. Healthcare providers may use the administered AGT dsRNA agent or As a measure of the effectiveness of AGT antisense polynucleotide agents, changes in the subject's blood pressure are monitored. It can be done.

[0164] A specific embodiment of the method of the present invention relates to one or more AGT-related diseases or conditions in a subject. Based at least in part on the evaluation of changes in the physiological characteristics of the number, the present invention's dsRNA agent or A This includes administering GT antisense polynucleotide agents to the target. For example, the present invention In one embodiment, the dsRNA agent or AGT anticeramic agent of the present invention is administered to the subject. The action of the sporonucleotide agent is determined and used to administer the dsR of the present invention to the subject. It can help adjust the dosage of NA agents or AGT antisense polynucleotide agents. In a non-limiting example, the dsRNA agent or AGT antisense polynucleotide of the present invention The drug Otide is administered to the subject, and the subject's blood pressure is determined after administration, and that determined level is... Based at least in part on the physiology of the administered drug, such as lowering or further lowering the target blood pressure. To enhance the scientific effect, use a larger amount of dsRNA agent or AGT antisense polynucleotide. It is determined whether or not a leotide agent is necessary. In another non-restrictive example, the dsRNA of the present invention When the agent or AGT antisense polynucleotide agent is administered to the subject, the subject's blood pressure after administration Determined, and based at least in part on that determined level, a smaller amount of dsR Administration of NA agents or AGT antisense polynucleotide agents is expected.

[0165] Therefore, in some embodiments of the present invention, the dsRNA of the present invention is administered to the subject. To adjust the dosage of the agent or AGT antisense polynucleotide agent, from before the treatment of the subject This includes evaluating changes in one or more physiological characteristics. Partial implementation of the method of the present invention The morphology is characterized by one, two, three, four, five, six or more physiological features of AGT-related diseases or conditions. The above determination; administered AGT dsRNA agent or AGT antisense polynucleotide of the present invention Evaluation and / or monitoring of the efficacy of otidol agents; and optionally, AGT-related aspects in the subjects. The present invention provides a dsRNA agent or AGT antisense polynucleotide for treating a disease or condition. Adjust one or more of the dosage, administration plan, and / or frequency of administration of leotide drugs. The use of the determined results; including. In some embodiments of the present invention, the subject is the present invention. The desired result can be achieved by administering an effective dose of a dsRNA agent or an AGT antisense polynucleotide agent. The result is that the subject's blood pressure decreases compared to the previously determined blood pressure for the subject. Or, blood pressure within the normal range.

[0166] When used herein in relation to AGT-related diseases or conditions, "treatment" means The terms "therapeutic" or "treated" refer to the subject who develops an AGT-related disease or condition. This can mean preventive treatment that reduces the likelihood of AGT and resolves AGT-related diseases or conditions. Therefore, or in order to reduce the level of AGT-related disease or condition, AGT-related disease or To prevent the disease from becoming more severe, and / or to increase the AGT polypeptide activity in the target population. Compared to subjects who have not received treatment to reduce AGT-related disease or illness, the subjects have AGT-related disease or illness. To slow the progression of the condition, treatment after the onset of AGT-related disease or symptoms is also important. It can taste good.

[0167] Specific embodiments of the activating agent, composition, and method of the present invention inhibit AGT gene expression. It can be used for the following purposes. With respect to the expression of the AGT gene, the term "inhibition" as used herein is used. "Harm," "silence," "reduction," "downregulate," and The term "knockdown" refers to the control level of RNA transcribed from the AGT gene, or expression level. The control level of AGT activity, or the control level of AGT translated from mRNA. In comparison with the present invention, the AGT dsRNA agent or AGT antisense polynucleotide Exposure of cells, groups of cells, tissues, organs, or subjects to a drug (e.g., treatment with it) ) In the case of the following, for example: cells, groups of cells, tissues, or organs in which the AGT gene is transcribed. or, in the subject, the level of RNA transcribed from the gene, the activity of expressed AGT Levels, and AGT polypeptides, proteins, or protein derivatives translated from mRNA. The development of the AGT gene by one or more of the following: a reduction in the level of bunits This means modifying the current state. In some embodiments, the control level is AGT d Not exposed to sRNA agents or AGT antisense polynucleotide agents (for example, A (Not treated with GT dsRNA agents or AGT antisense polynucleotide agents) This refers to the level in cells, tissues, organs, or objects.

[0168] Method of administration Various routes of administration for AGT dsRNA agents or AGT antisense polynucleotide agents It can be used in the method of the present invention. The selection of a specific delivery mode depends on the specific condition being treated and the treatment. It depends at least in part on the dosage required for therapeutic efficacy. Generally, the method of the present invention is medically It may be administered using any acceptable form of administration, and may cause clinically unacceptable side effects. Without causing any harm, it provides an effective level of treatment for AGT-related diseases or conditions. This means the form of. In some embodiments of the present invention, AGT dsRNA agent or AGT Antisense polynucleotides are administered orally, enterally, transmucosally, subcutaneously, and / or parenterally. It can be administered by route. The term "parenteral" refers to subcutaneous, intravenous, intrathecal, intramuscular, and abdominal. This includes intracavitary and intrasternal injection or infusion techniques. Other routes include, but are not limited to, transnasal injections (e.g., Examples include administration via a nasogastric tube, percutaneous, transvaginal, transrectal, sublingual, and inhalation. Delivery routes include intrathecal delivery, intraventricular delivery, or intracranial delivery. Partial embodiment of the present invention In terms of form, AGT dsRNA agents or AGT antisense polynucleotide agents are slow The substance is placed in an efficacy matrix and administered by positioning the matrix on the target. This is possible. In some aspects of the present invention, an AGT dsRNA agent or an AGT antisense agent can be used. Polynucleotide agents are nano-sized Particles can be used to deliver to target cells. Various delivery modes, methods, and reagents are included in this technology. It is publicly known in the field. Non-limiting examples of delivery methods and delivery agents are provided elsewhere in this specification. In some aspects of the present invention, an AGT dsRNA agent or an AGT antisense agent is used. With regard to dinucleotide drugs, the term "delivery" refers to one or more "bare" AGT ds The sequence of an RNA agent or AGT antisense polynucleotide agent is administered to cells or subjects. This may mean that. In certain aspects of the present invention, "delivery" means transfection Therefore, administer to cells or subjects, AGT dsRNA agents or AGT antisense agents. Delivering cells containing polynucleotide agents, or to cells and / or subjects, AGT Send a vector encoding a dsRNA agent or an AGT antisense polynucleotide agent. It means to reach. Transfection-based AGT dsRNA agent or AGT Delivery of antisense polynucleotide agents includes administration of the vector to cells and / or targets. It is visible.

[0169] In a part of the present invention, one or more AGT dsRNA agents or AGT AN Thicens polynucleotide preparations are available in pharmaceutical concentrations as salts, buffers, preservatives, and compatible materials. A preparation (preparation) that may generally contain a sexual carrier, an adjuvant, and optionally a therapeutic component. It can be administered in the form of (ation) or in a drug-acceptable solution. Part of the present invention In this embodiment, the AGT dsRNA agent or the AGT antisense polynucleotide agent , it can be formulated together with another therapeutic agent for simultaneous administration. According to the method of the present invention, AG T dsRNA agents or AGT antisense polynucleotide agents are administered in the form of a pharmaceutical composition. It is possible. Typically, pharmaceutical compositions are AGT dsRNA agents or AGT antisense agents. It comprises a polynucleotide agent and optionally a drug-acceptable carrier. The carriers are well known to those skilled in the art. The carriers used herein as pharmaceutically acceptable are This refers to the effectiveness of the biological activity of the active ingredient (for example, AGT gene expression in cells or subjects). (the ability of AGT dsRNA agents or AGT antisense polynucleotide agents to inhibit) This refers to non-toxic substances that do not interfere with other substances. dsRNA agents or AGT anticeres for therapeutic use. Various methods for administering and delivering spolynucleotides are known in the art, and the present invention It can be used in the following method.

[0170] Acceptable carriers for pharmaceuticals include diluents, fillers, salts, buffers, stabilizers, and solubilizers. and other substances known in the art are also examples. Acceptable carriers as exemplary agents are: As described in U.S. Patent No. 5,211,657, other carriers are known to those skilled in the art. Yes, such preparations generally contain salts, buffers, preservatives, compatible carriers, and optionally other therapeutic agents. It may contain [a certain substance]. When used in pharmaceuticals, the salt should be acceptable as a drug. However, salts that are not acceptable as pharmaceuticals can be conveniently used to prepare salts that are acceptable as pharmaceuticals. Such pharmacological and pharmaceutical products may be used and are not excluded from the scope of the present invention. The following acids are acceptable, but are not limited to: hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, and ricomethic acid. Prepared from nic acid, maleic acid, acetic acid, salicylic acid, citric acid, formic acid, malonic acid, succinic acid, etc. Salts that are used as drugs include sodium, potassium, and They can be prepared as alkali metal salts such as calcium salts or alkaline earth metal salts.

[0171] Some embodiments of the method of the present invention involve one or more AGT dsRNA agents or AGT This includes administering an antisense polynucleotide agent directly to tissue. In some embodiments, In this case, the tissue to which the compound is administered has an AGT-related disease or condition, or is experiencing These are tissues that can be affected, and non-limiting examples include the liver or kidneys. Direct tissue drugs Delivery can be achieved by direct injection or other means. Many of these compounds naturally enter and pass through the liver and kidneys, and are part of the therapeutic method of the present invention. The treatment includes oral administration of one or more AGT dsRNA agents. , or in combination with other therapeutic agents, AGT dsRNA agents or AGT antisense porin Cleotide drugs can be administered once or multiple times. When administered, AGT dsRNA agents or AGT antisense polynucleotide agents are different. It may be administered by the following route. For example, although not intended to restrict, the first (or most The initial doses may be administered subcutaneously, and one or more additional doses may be administered orally and / or systemically. It may be administered.

[0172] Systemic administration of AGT dsRNA agents or AGT antisense polynucleotide agents. With regard to embodiments of the present invention in which an AGT dsRNA agent or AGT antisense agent is desired, Linucleotide preparations are administered parenterally by injection, for example, by bolus injection or continuous infusion. It can be formulated for use. Injectable formulations may or may not contain preservatives. It may be available in unit dosage forms such as ampoules or multi-dose containers. AGT dsRNA preparations (medical (Also known as a pharmaceutical composition) is a suspension, solution, or emulsion in an oil or aqueous carrier. Formulation agents that can take the form of a suspension, stabilizer and / or dispersant (formu It may contain a latory agent. Dosage forms for parenteral administration include sterile aqueous solutions, non-aqueous solutions, suspensions, and emulsions. Examples of non-aqueous solvents include propylene glycol, polyethylene glycol, and olive oil. Examples include vegetable oils and injectable organic esters such as ethyl oleate. Suitable carriers include water, alcohol / aqueous solutions, emulsions, or suspensions, such as physiological saline. Examples include buffer media. Parenteral carriers include sodium chloride solution and Ringerbud. Examples include glycan solution, glucose and sodium chloride solution, lactated Ringer's solution, or fixed oil. It is used as an intravenous excipient. It is used as a liquid and nutritional supplement and electrolyte supplement (lingua). Examples include solutions based on glucose solution. Preservatives, antibacterial agents, and antioxidants. Other additives such as chelating agents and inert gases may also be present. Other forms of administration include intravenous administration. Depending on the patient's condition, the dosage may be reduced. If the patient's response to the initial dose is inadequate, the patient's tolerance may be reduced. Depending on the sex, a higher dose may be administered to the extent that is tolerable (or different, more localized) The dose can be effectively increased depending on the delivery route. One or more AGT dsRNA agents Or achieve appropriate systemic or local levels of AGT antisense polynucleotide agents. To achieve this, and to achieve an appropriate reduction in AGT protein activity, multiple doses per day are administered. The "give" can be used as needed.

[0173] In other embodiments, the method of the present invention involves biocompatible microparticles, nanoparticles, or other materials. This includes the use of a delivery carrier, such as an implant suitable for implantation in the recipient. An exemplary biodegradable implantable tablet that can be used according to the method is described in International Publication No. 95 / 24929. As described in the brochure (which is incorporated herein by reference), contains biomolecules. The biocompatible and biodegradable matrix required for this purpose is explained.

[0174] The target is one or more AGT dsRNA agents or AGT antisense polynucleotides. To deliver the drug, both non-biodegradable and biodegradable polymer matrices are used in this invention. It can be used in the method. In some embodiments, the matrix is ​​biodegradable The matrix polymer can be natural or synthetic polymer. The polymer is released during the period for which release is desired, typically ranging from a few hours to a year or more. The selection can be made based on the following criteria. Typically, the duration ranges from a few hours to 3 or 12 months. The output is available. The polymer optionally takes the form of a hydrogel and in water approximately its weight It can absorb up to 90% and can be further crosslinked optionally with polyvalent ions or other polymers. ru.

[0175] Typically, AGT dsRNA agents or AGT antisense polynucleotide agents are the present invention. In some embodiments, biodegradable implantable tablets are used to diffuse the polymer matrix or It can be delivered by decomposition. Exemplary synthetic polymers for this purpose are well known in the art. Known. Using biodegradable and non-biodegradable polymers, known in the art. The AGT dsRNA agent or AGT antisense polynucleotide agent is delivered using this method. It is possible to use AGT dsRNA agents or A for the treatment of AGT-related diseases or conditions. To deliver GT antisense polynucleotide agents, biodegradable (bioerodi ble) Bio-adhesive polymers such as hydrogels (HSSawhney, CPPa thak and JAHubellin Macromolecules,199 3, 26, 581-587) can also be used. Other suitable delivery systems include Examples include sustained-release, delayed-release, or sustained-release delivery systems. Such systems include AGT. Avoid repeated administration of dsRNA agents or AGT antisense polynucleotide agents. This could improve convenience for the target population and healthcare professionals. The types are available and known to those skilled in the art (e.g., U.S. Patent No. 5,075, U.S. Patent No. 109, U.S. Patent No. 4,452,775, U.S. Patent No. 4,675,18 U.S. Patent No. 9, U.S. Patent No. 5,736,152, U.S. Patent No. 3,854,480 U.S. Patent No. 5,133,974, and U.S. Patent No. 5,407,686 (See specifications). In addition, pump-type hardware delivery systems are available, and some of them It is also suitable for planting.

[0176] The use of long-term sustained-release implantable tablets is appropriate for patients with recurrent AGT-related disease or condition and It is suitable for the prophylactic treatment of at-risk subjects. As used herein, long-term release means The therapeutic level of AGT dsRNA agents or AGT antisense polynucleotide agents is low. Even if not available, up to 10 days, 20 days, 30 days, 60 days, 90 days, 6 months, 1 year, This refers to implantable tablets configured and positioned to deliver for a period of time of or longer. The locking mechanism is well known to those skilled in the art and includes part of the aforementioned release system.

[0177] A molecule or compound of the desired purity can be used as any agent, carrier, excipient, or agent. Fixed preparation [Remington's Pharmaceutical Sciences 2 By mixing with the 1st edition (2006), AGT dsRNA agent Alternatively, a therapeutic formulation of AGT antisense polynucleotide agents in the form of a lyophilized preparation or aqueous solution. It can be manufactured and used for storage in its current state. Acceptable carriers, excipients or stabilizers are used. It is nontoxic to the recipient at the dose and concentration it can tolerate, and: phosphoric acid, citric acid and Buffering agents such as other organic acids; antioxidants such as ascorbic acid and methionine; preservatives ( For example, stearyldimethylbenzylammonium chloride, hexamethonium chloride, benzylammonium chloride Inzalkonium, benzethonium chloride, phenol, butanol, and benzyl alcohol , or parabens such as methyl or propylparaben, catechol, resorcinol, Cyclohexanol, 3-pentanol, and m-cresol; low molecular weight (approximately 10 molecules) Peptides (full number of residues); proteins such as serum albumin, gelatin, or immunoglobulin; Hydrophilic polymers such as polyvinylpyrrolidone; glycine, glutamine, asparagine, hyaluronic acid Amino acids such as stidine, arginine, or lysine; monosaccharides, disaccharides, and glucose, mannor Other carbohydrates such as sucrose or dextrin; chelating agents such as EDTA; sucrose, manifold, etc. Sugars such as tol, trehalose, or sorbitol; salt-forming counterions such as sodium; Metal complexes (e.g., zinc-protein complexes); and / or TWEEN®, P LURONICS® or non-ionic polyethylene glycol (PEG) Contains a surfactant.

[0178] Cells, subjects, and controls The method of the present invention can be used with cells, tissues, organs and / or subjects. In some manifestations, the subject is humans or vertebrate mammals, but is not limited to dogs. These include cats, horses, cows, goats, mice, rats, and primates such as monkeys. Therefore, the present invention is used to treat AGT-related diseases or conditions in human and non-human subjects. It may be used. In some embodiments of the present invention, the subject is livestock, zoo animals, domesticated animals, or non- The method of the present invention can be used in veterinary prevention and treatment, and may apply to domesticated animals. In some embodiments of the present invention, the subject is a human, and the method of the present invention is a human pre- It can be used in both protective and therapeutic treatments.

[0179] A non-limiting example of a subject to which the present invention may apply is also called "high AGT expression levels." , diagnosed as a disease or condition accompanied by levels higher than the desirable AGT expression and / or activity levels. These are subjects who have been diagnosed, suspected of having, or are at risk of developing AGT. AGT expression and / Or, non-limiting cases of diseases and conditions involving levels of activity higher than the desired level, This is described elsewhere in the details. The method of the present invention is used at the time of treatment for a disease or medical condition. Subjects diagnosed with the disease, with a higher level of desirable AGT expression and / or activity. Subjects with a disease, or a disease with AGT expression and / or activity levels higher than the desired level. It can be applied to subjects who are considered to be at risk of suffering from or having a disease. In some aspects of the present invention In this context, a disease or condition characterized by AGT expression and / or activity levels higher than the desired level. The condition is an acute disease or medical condition; in certain embodiments of the present invention, AGT expression and / or activity A disease or condition characterized by a sexual level higher than the desirable level is a chronic disease or condition.

[0180] In non-restrictive examples, the AGT dsRNA agent of the present invention is used for hypertension, such as essential hypertension. Blood pressure, secondary hypertension, hypertensive emergencies (malignant hypertension and accelerated hypertension, acute hypertension, This drug is administered to patients diagnosed with pregnancy-related hypertension and treatment-resistant hypertension. The method of treatment is based on the diagnosis of having a disease or medical condition at the time of treatment, or the disease or condition This may apply to individuals who are considered to have developed or be at risk of developing the disease.

[0181] In another non-restrictive example, the renin-angiotensin-aldosterone system (RAAS) Diseases or disorders caused by or associated with the activation of RAAS, or when RAAS is not functioning In order to treat the symptoms or progression of the disease or disorder that is activated, according to the present invention, A GT dsRNA agents are administered. The term "angiotensinogen-related disease" is A This includes diseases, disorders, or medical conditions that benefit from reduced GT expression. Such diseases are generally high It is accompanied by blood pressure. An unrestricted example of angiotensinogen-related disease is hypertension, for example. , borderline hypertension (also known as prehypertension), essential hypertension (essential hypertension or idiopathic hypertension) Secondary hypertension (also known as idiopathic hypertension), isolated Sexual systolic or diastolic hypertension, pregnancy-related hypertension (eGO, pre-eclampsia, eclampsia, and postpartum hypertension) Pre-eclampsia, diabetic hypertension, treatment-resistant hypertension, refractory hypertension, paroxysmal hypertension, renal hemorrhage Renal hypertension (also known as renal hypertension), Goldblatt hypertension, intraocular hypertension, glaucoma Disorders, pulmonary hypertension, portal hypertension, systemic venous hypertension, systolic hypertension, unstable hypertension, Hypertensive heart disease, hypertensive nephropathy, atherosclerosis, arteriosclerosis, vascular disease (peripheral) Vascular diseases, diabetic nephropathy, diabetic retinopathy, chronic heart failure, cardiomyopathy, diabetic cardiomyopathy , glomerulosclerosis, aortic coarctation, aortic aneurysm, ventricular fibrosis, sleep apnea, heart failure (for example) (left ventricular systolic dysfunction), myocardial infarction, anguina, stroke, kidney disease (for example, pregnancy) Chronic kidney disease or diabetic nephropathy in the setting, renal failure (e.g., chronic renal failure), recognition Examples include intellectual disability (e.g., Alzheimer's disease) and systemic sclerosis (e.g., scleroderma nephropathy). In certain embodiments, AGT-related disorders include intrauterine growth restriction (IUGR) or One example is fetal growth restriction.

[0182] The cells to which this invention can be applied are in vitro, in vivo, and in vitro cells. The cells are present in the subject, in the culture and / or in the suspension, or in any other appropriate manner. The state or conditions in which the method of the present invention can be applied are: liver cells, liver Cells, cardiac cells, pancreatic cells, cardiovascular cells, kidney cells, or human and non-human mammalian cells, etc. It may be other types of vertebrate cells. In a particular embodiment of the present invention, the method of the present invention The cells to which it can be applied are healthy, normal cells that are not known to be affected cells. In certain embodiments of the present invention, the methods and compositions of the present invention are used for the liver, hepatocytes, and heart. Applicable to cells, pancreatic cells, cardiovascular cells, and / or kidney cells. Specific to the present invention In this embodiment, the control cells are normal cells, but the results of treated cells with disease or pathological conditions are shown. In certain environments, such as when compared to untreated cells with disease or disease symptoms, the disease or disease It should be understood that cells exhibiting this characteristic can also be used as control cells.

[0183] According to the method of the present invention, the level of AGT polypeptide activity is determined, and the AGT polypeptide The active level can be compared to a control level. The control is a predetermined value that can take many forms. It's possible. It could be a single cutoff value, such as the median or mean. Example For example, in a group having normal levels of AGT polypeptide and / or AGT polypeptide activity Furthermore, there is an increase in the level of AGT polypeptide and / or AGT polypeptide activity. In such a group, it can be established based on comparing the groups. Another non-restrictive example of a comparison group. This refers to AGT-related data for a population without one or more symptoms or diagnoses of a disease or medical condition. A population having one or more symptoms or diagnoses of a disease or medical condition, or the siR of the present invention The group of subjects that received siRNA treatment according to the present invention compared to the group of subjects that did not receive NA treatment. This can be a group. Typically, the control is a seemingly healthy, normal individual in an appropriate age group or a seemingly healthy individual. Based on healthy, normal cells. In addition to predetermined values, the control according to the present invention is parallel with the experimental material. It will be understood that this can be a sample of the material being tested. For example, in parallel with the experimental sample. Examples include samples from a control population or control sample produced by manufacturing for testing purposes. In some embodiments of the present invention, a control is exposed to the AGT dsRNA agent of the present invention. Includes cells or subjects that are not treated with the AGT dsRNA agent of the present invention. In that case, the control level of AGT polypeptide and / or AGT polypeptide activity. The subject is exposed to the AGT dsRNA agent or AGT antisense polynucleotide agent of the present invention. The level of AGT polypeptide and / or AGT polypeptide activity in the cells or subjects. It can be compared to Bell.

[0184] In some embodiments of the present invention, the control level is determined for the subject by AGT poly AGT peptides can be determined at the peptide level for the same subject at different time points. The levels are compared. In a non-limiting example, the level of AGT is the AGT treatment of the present invention. This is determined in biological samples taken from subjects who have not received medical treatment. The biological sample is a serum sample. The AGT poly(A) is determined in the sample taken from the subject. Peptide levels can serve as a baseline or control value for a given subject. In the therapeutic method of the invention, after administering the AGT dsRNA agent once or multiple times, one or multiple A number of additional serum samples may be taken from the subject, and subsequent AG in one or more samples T polypeptide levels can be compared to the control / baseline levels of the subject. This method is used to evaluate the onset, progression, or regression of AGT-related diseases or conditions in the subjects. For example, the AGT dsRNA agent or AGT antisense agent of the present invention can be applied to the target. The levels were higher than those collected from the same subject after administration of the renucleotide agent. The levels of AGT polypeptide in the baseline sample taken indicate an AGT-related disease or illness. Representing a receding state, the administered AGT of the present invention in the treatment of AGT-related diseases or conditions. This demonstrates the effectiveness of dsRNA agents.

[0185] In a particular embodiment of the present invention, AGT polypeptide and / or determined for the subject. One or more of the AGT polypeptide activity levels in the same subject It can serve as a control for subsequent comparison of ptide and / or AGT activity levels. This allows for the evaluation of changes from the "baseline" AGT polypeptide activity in the subject. This becomes possible. Therefore, when the initial value is used as a reference level for the target, Using the AGT polypeptide level and / or initial AGT polypeptide activity level, To reduce the level of AGT polypeptide and / or AGT polypeptide activity in the target. The capabilities of the methods and compounds of the present invention may be demonstrated and / or determined.

[0186] Using the method of the present invention, the AGT dsRNA agent and / or AGT antisense agent of the present invention Polynucleotide agents may be administered to the target. Such dsRNAi agents are shown in Table 1, for example. Double-chain AD00051~AD00122-19-2, AD00163-3, AV012 Includes 27~AVAV01257 and AV01711. In some embodiments, preferred Examples of suitable dsRNAi agents include double-stranded AD00158, AD00163, AD0 0163-3, AD00159, AD00290, AD00300 or AD00122 Examples include: In other embodiments, preferred dsRNAi agents include, for example, AD0 0158-1, AD00158-2, AD00163-1, AD00159-1, or A D00300-1 is an example. In some other embodiments, such dsRNAi agents and For example, double-strand variants, such as double-strand AD00158, AD00163, AD00 163-3, AD00159, AD00290, AD00300 or AD00122 A riant is one example. The efficacy of the administration and treatment of the present invention is obtained from a subject at a prior time. The pre-administration level of AGT polypeptide in the sampled serum, or the non-exposure control level (for example) It can be evaluated by comparing it with the level of AGT polypeptide in the control serum sample. Administration and treatment When treated, the level of AGT polypeptide in serum samples taken from the subject was low. At least 0.5%, 1%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70% It is reduced by %, 80%, 90%, 95%, or more. The level of AGT polypeptide and A It is understood that both the level of GT polypeptide activity and the level of AGT gene expression are related. In a particular embodiment of the present invention, the present invention is used in an amount effective for inhibiting AGT gene expression. This includes administering AGT dsRNA and / or AGT antisense agents to the target, This reduces the level of AGT polypeptide in the target, and also reduces the AGT polypeptide The level of activity is reduced.

[0187] Some embodiments of the present invention involve one or more biological samples taken from one or more subjects. The presence or absence of AGT polypeptide in the material, and / or the amount (referred to as level in this specification) This includes determining the effectiveness of the therapeutic method of the present invention. For example, using the method and composition of the present invention, the AGT dsRNA agent and / or A of the present invention AGT poly(AGT) The peptide levels can be determined. AGT polypeptide levels in serum samples after administration and treatment. Bell's reduction is a reserve of AGT polypeptide in serum samples taken from subjects at a previous point in time. Compared to the dose level, or to the non-exposed control level (e.g., AGT polyp in control serum samples) Compared to petit dot levels, at least 0.5%, 1%, 5%, 10%, 20%, 30% , 40%, 50%, 60%, 70%, 80%, 90%, 95% or more, target The level of effectiveness of the treatment given is indicated.

[0188] In some embodiments of the present invention, AGT-related diseases or medical conditions determined for the subject Physiological characteristics can be used as a control result, and the physiological characteristics of the same subject at different time points. The decision is compared to the control result. In an unrestricted case, blood pressure (and / or AGT disease) (or other physiological characteristics of the disease) are either present in subjects who have not received any AGT treatment of the present invention. The values ​​are measured and can be used as a baseline or control value for the target. In the treatment method, after administering an AGT dsRNA agent once or multiple times to the subject, blood pressure is measured. It is determined and compared to the control / baseline levels of the subject, respectively. Using such comparisons, It is possible to evaluate the onset, progression, or relapse of AGT-related diseases or conditions in the subjects. For example, the target may be the AGT dsRNA agent or AGT antisense polynucleotide of the present invention. Blood pressure that is higher than that measured in the same subject after administration of the drug, baseline blood pressure obtained from the subject Blood pressure indicates a regression of AGT-related disease or disorder, and is used in the treatment of AGT-related disease or condition. The effectiveness of administering the AGT dsRNA agent of the present invention is demonstrated.

[0189] In some embodiments of the present invention, one or more physiological characteristics of AGT-related diseases or disorders The values ​​determined for a subject are used as a control for subsequent comparison of the physiological characteristics of the same subject. It can serve as a value to assess changes in the physiological characteristics of the subject's "baseline." This makes it possible to obtain an initial physiological profile in an individual, The initial physiological profile measured as a control of the subject is measured, and the method of the present invention and Using compounds, the level of AGT polypeptide in an individual and / or AGT polypeptide It is possible to determine the effect of reducing the activity of butylene. Using the method of the present invention, AGT dsRNA agents and / or AGT antisense polynucleotide agents are AGT The present invention allows for administration to a target in an amount effective for treating a disease or medical condition. Effectiveness is determined by determining changes in one or more physiological characteristics of AGT disease or condition. It can be evaluated as follows. In an unrestrictive example, the blood pressure of the subject is the subject at a previous point in time. When compared to blood pressure obtained from, or to non-exposed control blood pressure, the subject's blood pressure is within the normal range. Until it reaches within the normal range, at least 0.5 mmHg, 1 mmHg, 2 mmHg, 3 mmHg, 4 mmHg mHg, 5mmHg, 6mmHg, 7mmHg, 8mmHg, 9mmHg, 10mmHg , 11mmHg, 12mmHg, 13mmHg, 14mmHg, 15mmHg, 16mm Reduced to Hg, 17mmHg, 18mmHg, 19mmHg, 20mmHg or higher. ru.

[0190] Some embodiments of the present invention, though not limited to them, include: (1) measuring the blood pressure of a subject; (2 ) Evaluate the physiological characteristics of one or more biological samples taken from one or more subjects. (3) Physical examination of the subject; using methods such as the presenting of AGT-related diseases or symptoms This includes determining the presence or absence of physical characteristics and / or changes thereof. This determination is made in accordance with the therapeutic method of the present invention. It can be used to evaluate the effectiveness of the law.

[0191] kit One or more AGT dsRNA agents and / or AGT antisense polynucleotides The present invention also includes a kit containing a drug, instructions for its use in the method of the present invention, and the drug itself. It is within the range. The kit of the present invention is useful in the treatment of AGT-related diseases or conditions. dsRNA agents, AGT sense polynucleotides, and AGT antisense polynucleotides May contain one or more of the following: one or more AGT dsRNA agents, AGT Kit containing sense polynucleotides and AGT antisense polynucleotide agents The kit of the present invention can be manufactured for use in the therapeutic method of the present invention. The components can be packaged in an aqueous medium or in a freeze-dried state. The kit of the present invention is one type or multiple types of container means or a series of container means (test tubes, vials, flasks, bottles, silicone May include a carrier separated for sealing in a container (such as a jar). First container means or a series of container means This includes one or more compounds, such as AGT dsRNA agents and / or AGT senses. The second container means or a series of container means may contain an antisense polynucleotide agent. AGT dsRNA and / or AGT administered in embodiments of the therapeutic method of the present invention Targeting agents, labeling agents, etc., which may be included as part of an antisense polynucleotide. Alternatively, it may contain a delivery agent or the like.

[0192] The kit of the present invention may also include instructions. Instructions are usually in written form and vary depending on the kit. Guidelines for implementing the treatment that is concretized and for making decisions based on that treatment. To provide.

[0193] The following examples are provided to illustrate specific examples of the implementation of the present invention and the scope of the invention. This invention is not intended to limit anything. The present invention is applicable to a variety of compositions and methods. It will be clear that this is the case. [Examples]

[0194] Example 1. Synthesis of RNAi agents The AGT RNAi double helix shown in Tables 2-4 above was synthesized following the basic procedure described below. : Using established solid-phase synthesis based on phosphoramidite chemistry, siRNA sense and A The nucleotide chain sequence was synthesized using an oligonucleotide synthesizer. Chain propagation is a four-step cycle: deprotection, condensation, capping. This was achieved using oxidation or sulfidation steps for the addition of each nucleotide. (Porous glass) Synthesis was carried out using a solid support made of (CPG, 1000 Å). Monomer phosphoramidites Purchased from the supplier. GalNAc ligand cluster (GLPA1 as an unrestricted example) A phosphoramidite having GLPA2 is prepared according to the procedure of Examples 2-3 of this specification. Synthesized. siRNA used for in vitro screening (Table 2) For this, synthesis was carried out on a 2 μmol scale; for in vivo testing, s For iRNA (Tables 3 and 4), synthesis was performed on a scale of 5 μmol or more. When an Ac ligand (GLO-0 as a restrictive example) is bound to the 3' end of the sense strand A CPG solid support to which a GalNAc ligand is bound was used. The c ligand (GLS-1 or GLS-2, for example, is linked to the 5' end of the sense strand) If combined, GalNAc phosphoramidite (as an unrestricted example, GLPA1 or GLPA2 was used in the final coupling reaction. Lichloroacetic acid (TCA) is used to deprotect the 4,4'-dimethoxytrityl protecting group (DMT). Used. 5-ethylthio-1H-tetrazole was used as an activator. THF I2 in / Py / H2O and phenylacetyl disulfide (P) in pyridine / MeCN ADS) was used in the oxidation reaction and the sulfidation reaction, respectively. After the final solid-phase synthesis step, The oligomer bound to the support body is cleaved, and a 1:1 volume 40 wt% methylamine aqueous solution is added. The protecting group was removed by treatment with a 28% ammonium hydroxide solution. The crude mixture was concentrated to synthesize siRNA for screening. The remaining solid was Dissolve in 1.0 M NaOAc, add ice-cold EtOH, and form a single chain as a sodium salt. The resulting product was precipitated and used in annealing without further purification. For in vivo testing. To synthesize siRNA, the crude single-stranded product is subjected to ion-pair reverse-phase HPLC (IP-RP). Further purification was performed by HPLC. Dissolve in 1.0 M NaOAc and ice-cold EtOH. By adding and precipitating, purified single-chain oligonucleotides can be obtained from IP-RP-HPLC. The ocidal product was converted to a sodium salt. Sense and antisense oligonucleotide chains were also used. Annealing is performed by equimolar complementation in water, and The main-chain siRNA product was formed and freeze-dried to obtain a cottony white solid.

[0195] Example 2. Preparation of intermediates A and B As shown in Scheme 1 below, commercially available galactose pentaacetic acid is used in dichloromethane (DCM). Treat samin with trimethylsilyltrifluoromethanesulfonate (TMSOTf). Intermediate A was synthesized by this process. Subsequently, Cbz-protected 2-(2-aminoethoxy Compound II was obtained by glycosylation with ethane-1-ol. Hydrogenation yielded Cbz The protecting group was removed to obtain intermediate A as a trifluoroacetic acid (TFA) salt. Cbz protection 2- (2-(2-aminoethoxy)ethoxy)ethane-1-ol plays the role of a starting material. Intermediate B was synthesized using the same scheme, except for one additional step. [ka]

[0196] Scheme 1 Compound I (20.0 g, 51.4 ml) in 100 mL of 1,2-dichloroethane (DCE) TMSOTf (17.1 g, 77.2 mmol) was added to a mol (mol) solution. The reaction solution was stirred at 60°C for 2 hours, and then stirred at 25°C for 1 hour. Cbz protection 2-( 2-aminoethoxy)ethane-1-ol (13.5g, 56.5mmol) is DCE( Dissolve in 100 mL, dry on a 4 Å powder molecular sieve (10 g), and under an N2 atmosphere. The reaction solution was added drop by drop at 0°C. The resulting reaction mixture was incubated at 25°C for 16 hours in N2. 2. Stirred under atmospheric conditions. Filter the reaction mixture and add saturated NaHCO3 (200 mL) and water ( The organic layer was washed with 200 mL of sodium hydroxide (Na2SO4) and saturated brine (200 mL). The mixture is dried, filtered and concentrated under reduced pressure to obtain the crude product, which is then converted to 2-methyltetrahydrochloride. The mixture was ground in Drofuran / heptane (5 / 3, v / v, 1.80 L) for 2 hours. The substance was filtered and dried to obtain compound II as a white solid (15.0 g, yield 50.3%). I obtained it.

[0197] Pour 10% Pd / C (1.50 g) into a dry and argon-purged hydrogenation flask. Add with caution, then 10 mL of tetrahydrofuran (THF), followed by 300 mL of THF. l) Solution of compound II (15.0 g, 26.4 mmol) and TFA (trifluoro Acetic acid (3.00 g, 26.4 mmol) was added. The resulting mixture was degassed and 3 times with H2. The mixture was purged and stirred at 25°C for 3 hours under an H2 (45 psi) atmosphere. Thin-layer chromatography Complete analysis of compound II by Raffy (TLC, solvent: DCM:MeOH = 10:1) Consumption was indicated. The reaction mixture was filtered under reduced pressure and concentrated. The residue was collected in anhydrous DCM (50 It was dissolved in 0 mL and concentrated. The process was repeated three times to obtain intermediate A as a foamy white solid ( It was obtained as 14.0 g, with a yield of 96.5%. 1 1H NMR (400MHz DMSO-d 6):δ ppm 7.90(d,J=9.29Hz,1H),7.78(br s,3 H),5.23(d,J=3.26Hz,1H),4.98(dd,J=11.29,3 .26Hz,1H),4.56(d,J=8.53Hz,1H),3.98-4.07( m,3H),3.79-3.93(m,2H),3.55-3.66(m,5H),2. 98(br d,J=4.77Hz,2H),2.11(s,3H),2.00(s,3 H), 1.90 (s, 3H), 1.76 (s, 3H).

[0198] Intermediate B was synthesized using a procedure similar to that used for the synthesis of intermediate A. 1 1H NMR (400M) Hz DMSO-d6):δ ppm 7.90(br d,J=9.03Hz,4H) ,5.21(d,J=3.51Hz,1H),4.97(dd,J=11.1Hz,1H ),4.54(d,J=8.53Hz,1H),3.98 -4.06(m,3H),3 .88(dt,J=10.9Hz,1H),3.76-3.83(m,1H),3.49 -3.61(m,9H),2.97(br s,2H),2.10(s,3H),1.9 9(s,3H),1.88(s,3H),1.78(s,3H).C 20 H 34 N2O1 Calculated mass of 1: 478.22; Measured mass: 479.3 (M+H + ).

[0199] Example 3. GalNAc ligand cluster phosphoramidites GLPA1, GLPA2 and synthesis of GLPA15 GLPAl and GLPA2 were prepared according to the following scheme 2. Benzyl protective agent Starting with pan-1,3-diamine, alkylation with t-butyl 2-bromoacetate Then, triester compound I was obtained. By removing the benzyl protecting group by hydrogenation, a secondary compound was obtained. Amine compound II was obtained by amide coupling with 6-hydroxycaproic acid. Compound III was obtained by treatment with HCl in dioxane. The t-butyl protecting group was then removed. Removed to obtain trioxidant compound IV. The amide cut of trioxidant compound IV with intermediate A or intermediate B was obtained. Pulling was performed to obtain compound Va or Vb. 2-Cyanoethyl N,N-diiso Using propyl chloride phosphoramidite and a catalytic amount of 1H-tetrazole, the compound By phosphorylating Va or Vb, phosphoramidite GLPA1 or GLPA 2 was combined. [ka]

[0200] Scheme 2 N-benzyl-1,3-propanedi in dimethylformamide (DMF, 100 mL) Add amine (5.00g, 30.4 mmol) and 2-t-butyl bromoacetate (23. Add to 7g, 121mmol) and then add diisopropylethylamine (DIEA, 2 3.61 g (182 mmol) was added drop by drop. The resulting reaction mixture was stored at 25-30°C. The mixture was stirred for 16 hours. From LC-MS, complete N-benzyl-1,3-propanediamine was found. Consumption was shown. The reaction mixture was diluted with H2O (500 mL) and ethyl acetate (500 mL) Extracted using (x2). The combined organic components were washed with saturated brine (1L). The solution is purified, dried with anhydrous Na2SO4, concentrated, and then concentrated under reduced pressure to obtain the crude product. This was subjected to silica gel column chromatography (gradient: petroleum ether:ethyl acetate 20:1~ It was purified by a 5:1 ratio. Compound I was obtained as a colorless oil (12.1 g, yield 78%). 0.4%). 1 H NMR(400MHz,CDCl3):δ ppm 7.26-7.4 0(m,5H),3.79(s,2H),3.43(s,4H),3.21(s,2H) ,2.72(dt,J=16.9,7.34Hz,4H),1.70(quin,J=7 0.2Hz, 2H), 1.44-1.50(m, 27H).

[0201] The dry hydrogenated bottle was purged three times with argon. Pd / C (200 mg, 10%) Add the following: MeOH (5 mL), then compound I (1.0) in MeOH (5 mL). (0g, 1.97 mmol) was added. The reaction mixture was degassed under vacuum and refilled with H2. This process was repeated three times. The mixture was heated at 25°C under an H2 (15 psi) atmosphere. The mixture was stirred for 12 hours. LC-MS showed that compound I was completely consumed. The mixture was filtered under reduced pressure in an N2 atmosphere. The filtrate was concentrated under reduced pressure to obtain a yellow oil. Compound II (655 mg, 79.7% yield) was obtained without further purification. It was then used in the next step. 1 ¹H NMR (400 MHz, CDCl3): δ ppm 3. 44(s,4H),3.31(s,2H),2.78(t,J=7.1Hz,2H),2 .68(t,J=6.9Hz,2H),1.88(br s,1H),1.69(qui n,J=7.03Hz,2H),1.44-1.50(s,27H).

[0202] Compound II (655 mg, 1.57 mmol), 6-hydroxy in DMF (6 mL) Caproic acid (249 mg, 1.89 mmol), DIEA (1.02 g, 7.86 mmol) l) 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDC I, 904 mg, 4.72 mmol) and 1-hydroxybenzotriazole (HOBt The mixture of 637 mg and 4.72 mmol was degassed, purged three times with N2, and then N2 The mixture was stirred at 25°C for 3 hours under controlled conditions. LC-MS revealed the target product. The mixture was diluted with H2O (10 mL) and extracted with 20 mL (10 mL x 2) of ethyl acetate. Combine the organic components, wash with saturated brine (20 mL), and dry with anhydrous Na2SO4. Filter and concentrate to obtain the crude product, which is then subjected to silica gel column chromatography (gradient). It is purified by petroleum ether:ethyl acetate (5:1~1:1) and compound III (650m g, yield 77.8%) was obtained as a yellow oil. 1 1H NMR (400MHz, CDCl) 3):δ ppm 3.90-3.95(s,2H),3.63(t,J=6.40Hz ,2H),3.38-3.45(m,6H),2.72(t,J=6.65Hz,2H) ,2.40(t,J=7.28Hz,2H),1.55-1.75(m,8H),1.4 4(s,27H).C 27 H 50 Calculated mass of N2O8: 530.36; Measured mass: 531 .3(M+H + ).

[0203] Compound III (5.5g, 10.3 mmol) in HCl / dioxane (2M, 55mL) The mixture of (1) was stirred at 25°C for 3 hours. LC-MS showed complete consumption of compound III. The reaction mixture was filtered, washed with alkyl(50 mL), and dried under reduced pressure. Crude product was obtained. This was dissolved in CH3CN (50 mL), and volatile components were removed under vacuum. This process was repeated three times to obtain compound IV as a white solid (2.05 g). , yield 54.5%. 1 H NMR(400MHz,D2O):δ ppm 4.21( s,1H),4.07(d,J=4.5Hz,4H),3.99(s,1H),3.45 -3.52(m,3H),3.42(t,J=6.5Hz,1H),3.32-3.38 (m,1H),3.24-3.31(m,1H),2.37(t,J=7.4Hz,1H ),2.24(t,J=7.4Hz,1H),1.99(dt,J=15.5,7.53 Hz,1H),1.85-1.94(m,1H),1.85-1.94(m,1H),1 .39-1.56(m,4H),1.19-1.31(m,2H).

[0204] Before stirring the mixture at 25°C under an N2 atmosphere for 3 hours, add DMF (10 mL) (496 Compound IV (500 mg, 1.05 mmol), intermediate A (3.67 mmol / mg) (2.02g,3.67mmol), DIEA(813mg,6.30mmol), ED The mixture of CI (704 mg, 3.67 mmol) and HOBt was degassed and perforated three times with N2. The desired product was shown by LC-MS. H2O (10 mL) was added. Therefore, the reaction mixture was quenched and extracted with DCM (10 mL x 2). The fraction was extracted with 10% citric acid (20 mL). The aqueous phase was neutralized with saturated NaHCO3 solution. Re-extraction was performed using CM (10 mL x 2). The organic components were dried with sodium sulfate and then under reduced pressure. The mixture was filtered and concentrated to obtain compound Va as a white solid (570 mg, 0.281 mmol). (26.8% yield). 1 H NMR:(400MHz,CDCl3)ppm δ 7.8 4-8.12(m,3H),6.85-7.15(m,2H),6.66-6.81(m ,1H),5.36(br d,J=2.7Hz,3H),5.11-5.27(m,3 H),4.63-4.85(m,3H),3.90-4.25(m,18H),3.37 -3.75(m,28H),3.15-3.28(m,4H),2.64(br d,J =6.53Hz,2H),2.30-2.46(m,2H),2.13-2.18(m, 9H),2.05(s,9H),1.94-2.03(m,18H),1.68(br s,2H),1.45(br s,2H),1.12(br t,J=7.0Hz,2H ).

[0205] In a solution of compound Va (260 mg, 0.161 mmol) in anhydrous DCM (5 mL), Following diisopropylammonium tetrazolide (30.3 mg, 0.177 mmol) 3-Bis(diisopropylamino)phosphanyloxypropanenitrile (194m (g, 0.645 mmol) was added dropwise at ambient temperature under N2. Reaction mixture 20 The mixture was stirred at ~25°C for 2 hours. LC-MS showed that compound Va was completely consumed. After cooling to -20°C, stir brine / saturated NaHCO3 (1:1, 5 mL) at 0°C. The reaction mixture was added to the mixed solution. After stirring for 1 minute, DCM (5 mL) was added to form a layer. This occurred. The organic layer was washed with brine / saturated NaHCO3 aqueous solution (1:1.5 mL). The solution was dried over Na2SO4, filtered, and concentrated to a volume of approximately 1 mL. Methyl t - The remaining solution was added drop by drop to 20 mL of butyl ether (MTBE). The result was... A white solid precipitated. The mixture was centrifuged and the solid was collected. The solid was redissolved in 1 mL of DCM. The mixture was dissolved and precipitated by adding MTBE (20 mL). The solid was then separated by centrifugation. It was isolated again. The recovered solid was dissolved in anhydrous CH3CN to remove volatile components. Repeat the process two more times to obtain the GalNAc ligand phosphoramidite compound GLP A1 (153 mg, 84.4 μmol) was obtained as a white solid. 1 1H NMR (400 MHz, CDCl3):ppm δ 7.71-8.06(m,2H),6.60-7. 06(m,3H),5.37(br d,J=3.0Hz,3H),5.18-5.32 (m,3H),4.70-4.86(m,3H),3.92-4.25(m,18H), 3.42-3.85(m,30H),3.25(m,4H),2.59-2.75(m, 4H),2.27-2.44(m,2H),2.15-2.20(s,9H)2.07( s,9H),1.96-2.03(m,18H),1.65(br s,4H),1.4 4(br d,J=7.28Hz,2H),1.14-1.24(m,12H). 31 P NMR (CDCl3): ppm δ 147.15.

[0206] Except for the use of intermediate-B, the same procedure was used to create a GalNAc ligand. We synthesized the sforamidite compound GLPA2. 1 1H NMR (400MHz, CDCl) 3):ppm δ 7.94-8.18(m,1H),7.69(br s,1H),6 .66-7.10(m,3H),5.35(d,J=3.5Hz,3H),5.07-5 .25(m,3H),4.76-4.86(m,3H),4.01-4.31(m,10 H),3.91-4.01(m,8H),3.74-3.86(m,4H),3.52- 3.71(m,30H),3.42-3.50(m,6H),3.15-3.25(m, 4H),2.52-2.70(m,4H),2.22-2.45(m,2H),2.15 -2.22(s,9H),2.06(s,9H),1.95-2.03(m,18H), 1.77(br s,2H),1.58-1.66(m,4H),1.40(m,2H) ,1.08-1.24(m,12H). 31 P NMR (CDCl3): ppm δ 1 47.12.

[0207] To manufacture GLPA15, we followed Scheme 3 below. [ka] Intermediate compound II (275g, 660mmol, 1) in dichloromethane (2.75L) To a solution of 0.00 equivalents, add triethylamine (133g, 1.32mol, 2.00 equivalents) Add the following, followed by Cbz-Cl (169g, 990mmol, 1.50 equivalents) drop by drop. The compound II was added. The reaction solution was stirred at 25°C for 2 hours, and as a result, compound II was completely removed by LC-MS. It was shown that the reaction solution was converted to a saturated NaHCO3 (800 mL) solution and a saturated The samples were washed sequentially with Japanese brine (500 mL), and the organic phase was dried with anhydrous Na2SO4. Filtration followed. After removing the drying agent, the filtrate was concentrated to a dry state. The crude product was then subjected to column chromatography. During the feeding phase (SiO2,PE / EA = 100 / 1 to 5 / 1), it transforms into a colorless oil. Compound 5 (290g, 527 mmol, yield 75.7%) was obtained. 1 1H NMR (400M) Hz in DMSO-d6):δ ppm 7.23-7.40(m,5H),5.0 0-5.12(m,2H),3.86-3.95(m,2H),3.23-3.39(m ,6H),2.55-2.67(m,2H),1.56-1.64(m,2H),1.3 1-1.46 (m, 27H).MS (ESI) [M+H] + m / z:551.6.

[0208] Add HCOOH (2.9 L) to compound 5 (145 g, 263 mmol, 1.00 equivalent). The compound was added, and the solution was stirred at 60°C for 12 hours. As a result, LC-MS showed complete transformation of compound 5. The exchange was shown. 1.5 L of toluene and acetnitrile toluene were added to the reaction solution, It was concentrated to approximately 500 mL under reduced pressure. Then, before concentrating to approximately 500 mL, toluene / Acetonitrile (1:1, approximately 750 mL) was added. Subsequently, the solution was concentrated to a dry state. Before proceeding, acetonitrile (approximately 1000 mL) was added. The crude product was heated to 60°C in acetonitrile. The material was finely ground in 700 mL of toril for 2 hours, then filtered. The solid was collected, dried, and white. A solid, compound 6 (105 g, quantitative analysis) was obtained. 1 1H NMR (400MHz in D) MSO-d6):δ ppm 7.26-7.40(m,5H),5.02-5.10( m,2H),3.89-4.00(m,2H),3.36-3.45(m,4H),3. 24-3.34(m,2H),2.59-2.72(m,2H),1.40(s,2H) .MS(ESI)[M+H] + m / z:383.0.

[0209] Compound 6 (100g, 261 mmol) and Intermediate A (502g, 915 mmol) Add TBTU (327g, 1.02mol) to a 1.0L solution of DMF (3.50 equivalents). (3.90 equivalents) and triethylamine (212 g, 2.09 mol, 8.00 equivalents) are added. The reaction was carried out at 25°C for 1 hour, and as a result, complete conversion of compound 6 was observed from LC-MS. The reaction solution was added to 4000 mL of water, and methyl t-butyl ether (2000 mL) was added. The remaining aqueous phase was extracted with dichloromethane (3000 mL). The dichloromethane phase was extracted using a 10% citric acid aqueous solution (2000 mL, washed twice). Saturated NaHCO3 (2.0 L, separated by two washes), and saturated bri (separated by two washes), and saturated bri The material was continuously washed with 2.0 L of water, and then dried with anhydrous sodium 2SO4. The filtrate was then drained under reduced pressure. Filtered and concentrated, a white solid, compound 8 (260g, 159mmol, yield 60.9%) was obtained. %) was obtained. 1 H NMR(400MHz in DMSO-d6):δ ppm 7. 99-8.08(m,2H),7.93(br d,J=5.50Hz,1H),7.7 9-7.86(m,3H),7.26-7.39(m,5H),5.22(d,J=3. 13Hz,3H),4.95-5.08(m,5H),4.54(br d,J=8.3 8Hz,3H),4.03(s,9H),3.81-3.93(m,5H),3.76( br d,J=4.88Hz,3H),3.44-3.62(m,10H),3.34- 3.43(m,6H),3.24(br d,J=6.13Hz,7H),3.02-3 .09(m,4H),2.40-2.47(m,2H),2.10(s,9H),1.9 9(s,9H),1.89(s,9H),1.77(s,9H),1.57-1.68( m,2H).MS(ESI)[M+H] + m / z:816.4.

[0210] Dry Pd / C (9g), followed by carefully adding MeOH (50mL) to moisten the Pd / C. Before proceeding, the 2L hydrogenation kettle was deactivated with argon. Next, the solution of compound 8 (90 (g, 55.1 mmol, 1.00 equivalent) and trifluorovinegar in MeOH (850 mL) The acid (6.29 g, 55.1 mmol, 1.00 equivalent) was slowly dissolved under an argon atmosphere. The mixture was degassed and replaced with H2 three times to obtain a hydrogen atmosphere. The mixture was stirred at 25°C for 10 hours. LC-MS showed the complete transformation of compound 8. Pd / C was removed by filtration, and the filtrate was concentrated under reduced pressure to obtain compound 9 (80g, yield 90%). 2% was obtained. 1 H NMR(400MHz in DMSO-d6):δ ppm 9 .12(br s,2H),8.50(br t,J=5.19Hz,1H),8.10 (br t,J=5.50Hz,2H),7.85-7.91(m,3H),5.22( d,J=3.25Hz,3H),4.95-5.01(m,3H),4.52-4.58 (m,3H),4.03(s,9H),3.84-3.93(m,3H),3.75-3 .83(m,3H),3.39-3.61(m,16H),3.23-3.32(m,6 H),3.15-3.18(m,3H),2.97-3.05(m,2H),2.54- 2.61(m,2H),2.10(s,9H),2.00(s,9H),1.89(s, 9H),1.77-1.80(m,9H),1.70-1.76(m,2H).MS(E SI)[M+H] + m / z:749.3.

[0211] Compound 9 (270g, 168mmol, 1.00 equivalent) of dichloromethane (2.7L) To the solution and glutaric acid anhydride (28.6 g, 252 mmol, 1.50 equivalents), triethyl Add ruamine (67.8g, 672 mmol, 4.00 equivalents) and leave the solution at 25°C for 1 hour. The mixture was stirred. LC-MS revealed that compound 9 was completely converted to compound 11. 4-hydroxypiperidine (42.4g, 420mmol, 2.50 equivalents) and TB Add TU (107g, 335 mmol, 2.00 equivalents) to the reaction solution and stir at 25°C. The process was continued for one hour. LC-MS showed the complete transformation of compound 11. Saturated NH4Cl(3) By slowly adding (0.0L), the reaction is quenched, the layers are separated, and the aqueous phase is separated. Extraction was performed using chloromethane (2 x 1000 mL) and combined with the previous organic phase. The combined organic phase was then... Wash with a 1:1 mixture of saturated NaHCO3 (aqueous solution) and saturated brine (3.0 L), then remove The solution was dried over sodium 2SO4, filtered under reduced pressure, and concentrated. It was then added to 1.5 L of dichloromethane. The crude product was dissolved and added dropwise to methyl t-butyl ether (7.5 L). During the addition process, a translucent white precipitate gradually formed. The precipitate was filtered under vacuum, and the solid was extracted. The mixture was recovered and dried under vacuum to obtain compound 13 as a white solid (207 g, yield 7 2.8%). 1 H NMR(400MHz in DMSO-d6):δ ppm 8. 05(br d,J=2.00Hz,2H),7.82(br d,J=7.38Hz, 3H),5.21(br s,3H),4.98(br d,J=10.26Hz,3H ),4.72(br s,1H),4.54(br d,J=7.88Hz,3H),4 .03(br s,9H),3.74-3.94(m,9H),3.45-3.71(m ,12H),3.40(br s,6H),3.24(br s,7H),3.07(b rd,J=14.13Hz,5H),2.91-3.01(m,1H),2.24-2 .44(m,5H),2.20(br s,1H),2.10(s,9H),1.96- 2.04(m,9H),1.89(br s,9H),1.74-1.81(m,9H) ,1.51-1.73(m,6H),1.07-1.36(m,3H).MS(ESI) [M+H] + m / z:848.0.

[0212] Compound 13 in dichloromethane (2.0 L) (200 g, 118 mmol, 1.00 equivalent) (Quantity) and tetrazole diisopropylammonium (8.08 g, 47.2 mmol, 0 Add 3-bis(diisopropylamino)phosphanyloxypropane to a 0.40 equivalent solution. Nitrile (53.3 g, 177 mmol, 1.50 equivalents) was added. The reaction solution was heated at 40°C. The mixture was stirred for 2 hours, and as a result, LC-MS showed the complete transformation of compound 13. The reaction solution was washed with a 1:1 mixture of sodium NaHCO3 and saturated brine (2.0 L), and then anhydrous water was added. The product was dried with Na2SO4. After concentrating the filtrate, the crude product obtained was treated with dichloromethane (1 Dissolve in 0.2L of methyl t-butyl ether (6.0L) and add dropwise to the stirred methyl t-butyl ether. The suspension was filtered, and the filtered cake was rinsed with t-butyl ether. The solid was recovered and vacuum was applied. The mixture was dried below ground. The product was dissolved in dichloromethane (1.0 L) and concentrated to a dry state. Repeat this process four times to remove any remaining t-butyl ether and ensure compliance with GLPA15(1 64g was obtained, with a yield of 73.3%. 1 1H NMR (400MHz in DMSO-d6) ):δ ppm 8.05(br d,J=6.50Hz,2H),7.81(br d ,J=9.01Hz,3H),5.22(d,J=3.25Hz,3H),4.98(d d,J=11.26,3.25Hz,3H),4.55(br d,J=8.50Hz, 3H),4.03(s,9H),3.64-3.97(m,12H),3.55-3.6 3(m,6H),3.50(br s,5H),3.40(br d,J=6.13Hz ,6H),3.17-3.30(m,9H),3.07(br d,J=14.26Hz ,4H),2.76(t,J=5.82Hz,2H),2.18-2.47(m,6H) ,2.10(s,9H),1.99(s,9H),1.89(s,9H),1.78(s ,9H),1.52-1.74(m,6H),1.12-1.19(m,12H). 31 P NMR(DMSO-d6):ppm δ 145.25.MS(ESI)[M+H] + m / z:1895.7.

[0213] In some studies, GalNAc (GalNAc in this specification) was found at the 5' end of the sense strand. A method for attaching a targeting group containing a NAc delivery compound (also known as a NAc delivery compound), wherein the sense chain has 5 'Oligonoid chain extension to attach it to the end (i.e., the 5' end of the sense strand) Solid-phase synthesis using synthetic processes such as those used for the addition of nucleotides to the ends. Use of GalNAc phosphoramidite (GLPA1) in the final coupling process A method including this is provided.

[0214] Some studies have explored methods for attaching a GalNAc-containing targeting group to the 3' end of a sense chain. This includes the use of GLO-n-containing solid carriers (CPGs). In some studies, the sense strand A method for attaching a GalNAc-containing targeting group to the 3' end is to use an ester bond to solidify the CPG. Linking a GalNAc targeting group to the body support, during the synthesis of the sense chain, the GalNAc target The process involves using the resulting CPG to which the chemical group is attached, and as a result, the 3' end of the sense chain A GalNAc targeting group is obtained with the end attached. Other GalNAc phosphoramidites Compound (GLPAn) is similar to the method described herein, or is well known in the art. It can be obtained using known methods after the use of a reasonably equivalent intermediate, and standard It can be linked to the appropriate position on the siRNA double helix as a targeting group.

[0215] Example 4. In vitro screening of AGT siRNA double-stranded cells Hep3B cells were trypsin-treated, adjusted to the appropriate density, and then plated in a 96-well plate. Seeding was performed. Simultaneously with seeding, lipofectamine was applied according to the manufacturer's recommendations. Test siRNA using RNAiMax (Invitrogen-13778-150) Alternatively, control siRNA was transfected into cells. siRNA was administered at two different concentrations (0.2 The test was performed in triplicates at nM or 1.0nM, and the results were 4.6pM~1 The control siRNA was tested in sets of three at eight different concentrations using sequential 3-fold dilutions of 0 nM.

[0216] 24 hours after transfection, remove the culture medium and collect the cells for RNA extraction. Collected according to the manual. TRIzol (trademark) reagent (Invitrogen-155 Total RNA was extracted using 96018).

[0217] Follow the manual for the PrimeScript® RT reagent kit and gDNA E Raser (Perfect Real Time) (TaKaRa-RR047A) cDNA was synthesized using the following method. AGT cDNA was detected by qPCR. GAP DH cDNA was detected in parallel with the internal control. PCR was performed as follows: 40 cycles of 30 seconds at 95°C, followed by 10 seconds at 95°C and 30 seconds at 60°C.

[0218] Data Analysis Expression in each sample was quantified relatively (RQ) using the comparative Ct (ΔΔCt) method. It was decided that this method involves target genes and housekeeping genes (GAPDH) The difference in Ct (ΔCt) is measured.

[0219] The equation is shown below. ΔCT=Target gene average Ct-GAPDH average Ct ΔΔCT = ΔCT(sample) - ΔCT(random control or lipofectamine RNAiMa) (x comparison) Relative quantification of target gene mRNA = 2( -ΔΔCT ) Inhibition percentage = (Relative quantification of control - Relative quantification of sample) / Relative quantification of control × 100%

[0220] [Table 30]

[0221] [Table 31]

[0222] Example 5. In vivo testing of AGT siRNA double helix To evaluate the in vivo activity of AGT siRNA, the human AGT gene encoding A Mice infected with AV were used (4 mice per group). 14 days after siRNA administration... , 1 × 10⁻¹⁶ adeno-associated virus 8 (AAV8) vector encoding the human AGT gene 11 Female C57BL / 6J mice were infected by intravenous injection of virus particles. On day 0, AGT siRNA or PBS 2.5 mg / kg or 3 mg / kg was administered. A single dose was administered by subcutaneous injection. Blood samples were taken on day 0, before siRNA administration, and at the end of day 7. Collected. Manufacturer's recommended protocol (IBL America, Human Angiotensinus) Human AGT protein concentration by ELISA assay according to the ELISA kit The following was measured: Human in the liver of mice on day 7 between the siRN-treated group and the PBS-treated group. AGT mRNA levels (determined by qPCR) or human AGT levels in plasma samples The knockdown percentage was calculated by comparing the protein levels. The results are shown in Tables 6-9.

[0223] [Table 32]

[0224] [Table 33]

[0225] [Table 34]

[0226] [Table 35]

[0227] [Table 36]

[0228] Example 6. In vivo testing of AGT siRNA double-stranded cells To evaluate the in vivo activity of AGT siRNA, a total of 15 cynomolgus monkeys (13 We recruited animals (up to 22 years old, weighing 7-9 kg) for this study. Three animals were assigned to five different groups. The material was randomly divided, and 2 mg / kg of the test substance was subcutaneously injected into each animal. The quality is as follows: Compounds in Table 4 (AD00158-1, AD00158-2, AD00163 This corresponds to -1, AD00159-1, and AD00300-1.

[0229] After fasting overnight, -14th day (before administration), -7th day (before administration), 1st day (before administration) And on the 8th, 15th, 22nd, 29th, 43rd, 57th, 64th, and 7th days after administration. Blood samples were collected on days 1, 78, 85, and 92. Subsequently, the collected blood samples... Leave the mixture at room temperature for at least 30 minutes to allow it to solidify, then run it at 350 rpm at 4°C for 10 minutes. The serum was centrifuged. The recovered serum (approximately 1.0 mL) was placed into two pre-labeled bottles. Polypropylene screw-cap vials (0.5 ml / vial, one for ELISA assay, also One bottle was transferred to another (for later use) and stored in a -80°C freezer until testing.

[0230] Serum AGT protein levels were determined by ELISA. (Monkeys on day 1) Figure 1 shows the percentage remaining compared to the AGT level in plasma.

[0231] Example 7. In vitro screening of AGT siRNA double-stranded cells In vitro studies were conducted according to the method of Example 4, and the experimental results are shown in Table 10.

[0232] [Table 37]

[0233] Example 8. In vivo analysis of AD00163-3 in an AAV mouse model. test: After an adaptation period, 12 female C57BL / 6J mice were divided into two groups based on body weight: model (solvent). The mice were randomly divided into two groups: the ) group and the AD00163-3 (1 mg / kg) group. On day 1, each mouse AAV-AGT virus 1 x 10 in the tail vein 11 Inject vg, 100 μL / animal volume An animal model was established by injecting PBS or on day 15, as shown in Table 4. AD00163-3 was administered to mice in each group by subcutaneous injection at a volume of 5 mL / kg. It was administered by the following method. Before administration on day 15, blood was collected from the submandibular vein of each mouse. Serum samples were collected after centrifugation. On day 22, all mice were sacrificed with CO2, and the heart was extracted. Whole blood was collected by puncture, and serum samples were collected after centrifugation. Manufacturer's recommended protocol. (According to the IBL America Human Angiotensinogen ELISA Kit) Human AGT protein concentrations were measured by the LISA assay. The siRN treatment group and P Human-derived AGT protein levels in mouse plasma samples were compared on day 7 between the BS treatment groups. By doing so, the knockdown percentage was calculated. From that data, AD00 Treatment with 163-3 (1 mg / kg) increased the expression of human AGT protein in mouse serum. It was found that this could be significantly reduced by 91%.

[0234] Example 9. Testing of AD00163-3 in a spontaneous hypertension model in cynomolgus monkeys. Ten cynomolgus macaques with hypertension were randomly assigned to two groups (5 monkeys in each group). The doses were divided and administered at a rate of 10 mg / kg of physiological saline or AD00163-3 as shown in Table 4. - Day 6 and - Day 2 (before administration), and 2, 7, 14, 21, 28 and 35 days after administration. A blood sample was taken from the eye. Serum AGT concentration was measured using ELISA according to the manufacturer's recommended protocol. Blood pressure was measured using a tailcuff device. As shown in Figures 2 and 3, serum AGT was reduced (98% reduction on day 35 after administration), and at the same time, AD00163-3 10 mg / kg was administered subcutaneously as a single dose, resulting in a 28 m³ of activity on day 35 after administration. SBP in mHg decreased significantly (SBP changed to a baseline of 147-119 mmHg). (The change occurred), and there was no significant change in SBP in the control group during the same period (SBP was 14 (Changed to baseline of 4-145 mmHg). Mean arterial pressure and diastolic pressure (MBP and D) A significant decrease in BP was also confirmed, as shown in Figures 4 and 5, respectively.

[0235] Equivalents Although several embodiments of the present invention are described and illustrated herein, To perform the functions described and / or to obtain the results and / or one or more advantages. Therefore, various other means and / or structures may be used, as well as modifications thereof. It is readily apparent to those skilled in the art that each of these modifications is considered to fall within the scope of the present invention. It should be understood. More generally, all parameters, dimensions, materials, and The configuration is illustrative, and the actual parameters, dimensions, materials, and / or spatial arrangement are not included. It is readily apparent to those skilled in the art that the placement will vary depending on the specific application in which the teachings of this invention are used. This will be understood. Not only ordinary experiments, but also many of the specific embodiments of the present invention described herein Those skilled in the art will recognize or understand the use of an equivalent. Therefore, the embodiments described above are shown merely as examples and are not part of the attached claims. And within the scope of such equivalents, it is understood that other actions may be taken in addition to those specifically described and requested. The present invention relates to each of the individual features, systems, articles, materials and / or described herein. Regarding methods. Furthermore, two or more such features, systems, articles, materials and / or methods. Any combination of such features, systems, articles, materials and / or methods is inconsistent with each other. Provided that it does not fall within the scope of the present invention, it is included within the scope of the present invention.

[0236] All definitions defined and used herein are compiled by comparative dictionary definitions, references, etc. The definitions in the document and / or the terms defined are understood as having their usual meanings. It should.

[0237] If no quantitative limitations are used in this specification and in the claims, this is equivalent to Conversely, unless explicitly stated otherwise, it should be understood as "at least one."

[0238] As used herein and in the claims, the phrase "and / or" refers to an element The "one or both" of these is combined in this way, that is, when such elements are in a particular case It should be understood that this means they appear to have been combined separately in one case and in other cases. It is. In addition to the elements specifically identified by "and / or", the elements specifically identified Whether related to the element or not, unless explicitly specified otherwise, Other elements may exist at will.

[0239] All references, patents, patent applications, and publications described in or referenced in this application are the same as those of the original application. The entire text is incorporated herein by reference.

Claims

1. Double-stranded ribonucleic acid (dsRNA) drugs that inhibit the expression of angiotensinogen (AGT) The dsRNA agent comprises a sense strand and an antisense strand, and the antisense strand contains The nucleotides at positions 2-18 contain a region complementary to the AGT RNA transcript. The complementary region is one of the antisense sequences shown in Tables 1 to 4, and 0, 1, 2, Or it contains at least 15 neighboring nucleotides that are different from three nucleotides, and can be optionally targeted. A dsRNA agent containing a ligand.

2. The region that complements the AGT RNA transcript is one of the antisensors shown in Tables 1-4. One of the sequences and at least 15, 16, 17, 18 that differ by three or fewer nucleotides, Or the dsRNA agent according to claim 1, comprising 19 adjacent nucleotides.

3. The antisense strand of the dsRNA is less than any of the target regions of SEQ ID NO:

519. Claim 1 or that is substantially complementary and provided in any one of Tables 1 to 4 The dsRNA agent described in 2.

4. The antisense strand of the dsRNA is less than any of the target regions of SEQ ID NO:

519. The claims described in claim 3, which are at least completely complementary and provided in any one of Tables 1 to 4. dsRNA agent.

5. The dsRNA agent comprises a sense strand sequence described in any one of Tables 1 to 4, and the sense strand sequence is described in Table 1 to 4. The s-chain sequence is at least substantially different from the antisense chain sequence in the dsRNA agent. A dsRNA agent according to claim 1, which is complementary in nature.

6. The dsRNA agent comprises a sense strand sequence described in any one of Tables 1 to 4, and the sense strand sequence is described in Table 1 to 4. The strand sequence is completely complementary to the antisense strand sequence in the dsRNA agent. A dsRNA agent according to claim 1.

7. The dsR according to claim 1, comprising the antisense strand sequence described in any one of Tables 1 to 4. Na agent.

8. The dsR according to claim 1, which includes the sequence shown as a double-stranded sequence in any one of Tables 1 to 4. Na agent.

9. The dsRNA agent according to claim 1, comprising at least one modified nucleotide.

10. All or substantially all nucleotides in the antisense strand are modified nucleotides A dsRNA agent according to claim 1, which is a rheotide.

11. The at least one modified nucleotide is: 2'-O-methylnucleotide, 2'-F Luoronucleotide, 2'-deoxynucleotide, 2',3'-seconucleotide mimi Locked nucleotides, unlocked nucleic acid (UNA) nucleotides, Glycol nucleic acid (GNA) nucleotides, 2'-F-arabinonucleotides, 2'-meth Xyethyl nucleotide, debasalized nucleotide, ribitol, inverted nucleotide, inverted Base nucleotide, inverted 2'-OMe nucleotide, inverted 2'-deoxynucleotide, 2'-amino modified nucleotide, 2'-alkyl modified nucleotide, morpholino nucleo Tide and 3'-OMe nucleotide, nucleotide containing a 5'-phosphorothioate group, or a terminal nucleus linked to a cholesterol derivative or a dodecanoic acid bisdecylamide group Rheotides, 2'-amino modified nucleotides, phosphoramidates, or non-natural base-containing nucleotides A dsRNA agent according to claim 5 or 6, comprising a creotide.

12. Claim 9 or otherwise, the E-vinylphosphonate nucleotide is included in the 5' end of the guide chain. This is the dsRNA agent described in 10.

13. The d according to claim 1, comprising at least one phosphorothioate nucleoside interbonding. sRNA agent.

14. The sense chain includes at least one phosphorothioate nucleoside bond. The dsRNA agent described in item 1.

15. The antisense chain contains at least one phosphorothioate nucleoside bond. The dsRNA agent according to claim 1.

16. The sense chain is between 1, 2, 3, 4, 5, or 6 phosphorothioate nucleosides. A dsRNA agent according to claim 1, comprising binding.

17. The antisense chain comprises 1, 2, 3, 4, 5, or 6 phosphorothioate nucleos. A dsRNA agent according to claim 1, comprising intersidal binding.

18. All or substantially all nucleotides of the sense and antisense strands are modified. A dsRNA agent according to claim 1, wherein the agent is a nucleotide.

19. The modified sense chain is a modified sense chain sequence shown in one of Tables 2 to 4, according to claim 1. dsRNA agent.

20. The modified antisense chain is one of the modified antisense chain sequences shown in Tables 2 to 4. The dsRNA agent described in item 1.

21. The sense chain is complementary or substantially complementary to the antisense chain, and The dsRN according to claim 1, wherein the complementary region has a length of 16 to 23 nucleotides. Agent A.

22. The ds according to claim 21, wherein the complementary region has a length of 19 to 21 nucleotides. RNA agent.

23. The dsRNA agent according to claim 1, wherein each chain has a length of 30 nucleotides or less.

24. The dsRNA agent according to claim 1, wherein each chain has a length of 25 nucleotides or less.

25. The dsRNA agent according to claim 1, wherein each chain has a length of 23 nucleotides or less.

26. It contains at least one modified nucleotide and one or more targeting groups or links. The dsRNA agent according to claim 1, further comprising a group.

27. The one or more of the targeting groups or linking groups are conjugated to the sense chain. , the dsRNA agent according to claim 26.

28. The targeting group or linking group contains N-acetyl-galactosamine (GalNAc), A dsRNA agent as described in item 26 or 27.

29. Each targeting group has the following structure: 【Chemistry 1】 【Chemistry 2】 【Transformation 3】 【Chemistry 4】 【Transformation 5】 【Transformation 6】 【Transformation 7】 【Transformation 8】 【Chemistry 9】 【Chemistry 10】 【Chemistry 11】 A dsRNA agent according to claim 26 or 27, having the following characteristics.

30. The d according to claim 1, comprising a targeting group conjugated to the 5' end of the sense chain. sRNA agent.

31. The d according to claim 1, comprising a targeting group conjugated to the 3' end of the sense chain. sRNA agent.

32. The ds according to claim 1, wherein the antisense chain includes an inverted debase residue at its 3' end. RNA agent.

33. The sense strand contains one or two inverted debase residues at its 3' and / or 5' ends. , the dsRNA agent according to claim 1.

34. A dsRNA agent according to claim 1, comprising two blunt ends.

35. Claims that at least one strand comprises a 3' overhang of at least one nucleotide. The dsRNA agent described in 1.

36. Claims that at least one strand comprises a 3' overhang of at least two nucleotides. The dsRNA agent described in 1.

37. A composition comprising a dsRNA agent according to any one of claims 1 to 36.

38. The composition according to claim 37, further comprising a carrier that is acceptable as a pharmaceutical agent.

39. The composition according to claim 38, further comprising one or more additional therapeutic agents.

40. Packaging for kits, containers, packs, dispensers, pre-filled syringes or vials The composition according to claim 39, as described.

41. Claim 37, which is formulated for subcutaneous administration or for intravenous (IV) administration. The composition described.

42. A cell comprising a dsRNA agent according to any one of claims 1 to 36.

43. The cell according to claim 42, which is a mammalian cell, or optionally a human cell.

44. (i) A double-stranded ribonucleic acid (dsRNA) agent according to any one of claims 1 to 36 or To prepare cells containing an effective amount of the composition according to any one of claims 37 to 41. A method for inhibiting AGT gene expression in cells, including [the specified method].

45. (ii) The cells prepared in (i) of claim 44, the mRNA of the AGT gene Maintain the transcript for a sufficient time to obtain degradation of the AGT gene in the cells. The method according to claim 44, further comprising inhibiting the expression of the gene.

46. The claim is that the aforementioned cells are present in the subject and the dsRNA agent is administered subcutaneously to the subject. The method described in paragraph 44.

47. The claim is that the aforementioned cells are present in the subject and the dsRNA agent is administered intravenously to the subject. The method described in paragraph 44.

48. After administering the dsRNA agent to the subject, the inhibition of the AGT gene is evaluated. A method further including, (i) One or more physiological characteristics of the AGT-related disease or condition in the subject To decide, (ii) Baseline pretreatment physiological characteristics and / Or, the physiological characteristics of the control of the aforementioned AGT-related disease or condition, determined physiological This includes comparing specific characteristics, From the above comparison, one or more of the following can be determined: whether or not the expression of the AGT gene is inhibited in the subject. The method according to claim 46 or 47, wherein the following is shown.

49. The aforementioned physiological characteristics include essential hypertension, secondary hypertension, hypertensive emergency (malignant hypertension and This includes acute hypertension (such as accelerated hypertension), pregnancy-related hypertension, and treatment-resistant hypertension. The method according to claim 48, wherein the patient has hypertension.

50. Claim 49 states that a decrease in the target's blood pressure indicates a decrease in AGT gene expression in the target. Method of description.

51. A method for inhibiting AGT gene expression in a target, any one of claims 1 to 36. A double-stranded ribonucleic acid (dsRNA) agent as described in paragraph 1 or any one of claims 37 to 41 A method comprising administering the composition described above to the subject in an effective amount.

52. The method according to claim 51, wherein the dsRNA agent is administered subcutaneously to the subject.

53. The method according to claim 51, wherein the dsRNA agent is administered intravenously to the subject.

54. After administering the dsRNA agent to the subject, the inhibition of the AGT gene is evaluated. A method further including, (i) One or more physiological characteristics of the AGT-related disease or condition in the subject To decide, (ii) Baseline pretreatment physiological characteristics and / Or, the physiological characteristics of the control of the aforementioned AGT-related disease or condition, determined physiological This includes comparing specific characteristics, From the above comparison, one or more of the following can be determined: whether or not the expression of the AGT gene is inhibited in the subject. The method according to any one of claims 51 to 53, wherein the following is shown.

55. The aforementioned physiological characteristics include essential hypertension, secondary hypertension, hypertensive emergency (malignant hypertension and This includes acute hypertension (such as accelerated hypertension), pregnancy-related hypertension, and treatment-resistant hypertension. The method according to claim 54, wherein the patient has hypertension.

56. Claim 55, wherein a decrease in the target blood pressure indicates a decrease in AGT gene expression in the target. Method of description.

57. A method for treating diseases or conditions associated with AGT protein, wherein AGT gene expression To inhibit double-stranded ribonucleic acid (dsRN) according to any one of claims 1 to 36 A) Inject an effective amount of the agent or the composition described in any one of claims 37 to 41 into the subject. A method that includes giving in to something.

58. The aforementioned disease or condition is related to the activity of the renin-angiotensin-aldosterone system (RAS). Symptoms caused by, or related to, sexualization, or in response to RAAS activation Or progression, and the aforementioned diseases or conditions are not limited to the following hypertensive diseases, hypertension Borderline hypertension, essential hypertension, secondary hypertension, isolated hypertension, systolic or diastolic hypertension High blood pressure, pregnancy-related hypertension, diabetic hypertension, treatment-resistant hypertension, refractory hypertension, paroxysmal Hypertension, renovascular hypertension, Goldblatt hypertension, intraocular hypertension, glaucoma, pulmonary hypertension, Portal hypertension, systemic venous hypertension, systolic hypertension, unstable hypertension, hypertensive heart disease, Hypertensive nephropathy, atherosclerosis, arteriosclerosis, vascular disease, diabetic nephropathy, diabetic Retinopathy, chronic heart failure, cardiomyopathy, diabetic cardiomyopathy, glomerulosclerosis, aortic stenosis, aortic aneurysm, Ventricular fibrosis, heart failure, myocardial infarction, anguina, stroke, kidney disease, renal failure, systemic sclerosis, children Hypertension and one or more of the following conditions: Miyauchi growth restriction (IUGR) and fetal growth restriction. The method according to claim 57, which is usually related.

59. The method according to claim 57, further comprising administering an additional treatment regimen to the subject.

60. The aforementioned additional treatment regimen is one or more types of AGT antisense polynucleotides of the present invention Administering otide to the subject, administering a non-AGT dsRNA therapeutic agent, and The method according to claim 59, comprising performing a dynamic modification on the target.

61. The aforementioned non-AGT dsRNA therapeutics include the following: additional therapeutics, e.g., diuretics, angiotherapy Tensin-converting enzyme (ACE) inhibitors, angiotensin II receptor antagonists, β-blockers Drug discontinuation, vasodilators, calcium channel blockers, aldosterone antagonists, alpha -2 agonists, renin inhibitors, alpha-blockers, peripherally acting adrenergic agonists, selective D1 Receptor partial agonist, non-selective α-adrenergic antagonist, synthetic steroidal antimineral corticosteroid It is formulated as a ticoid, or any combination of the above, and a combination of agonists. The method according to claim 60, wherein the method is one or more of the drugs used to treat hypertension.

62. The method according to claim 57, wherein the dsRNA agent is administered subcutaneously to the subject.

63. The method according to claim 57, wherein the dsRNA agent is administered intravenously to the subject.

64. The efficacy of the administered double-stranded ribonucleic acid (dsRNA) agent in the subject is determined. The method according to any one of claims 57 to 63, further comprising:

65. The method for determining the effectiveness of treatment in the aforementioned subjects is: (i) One or more physiological characteristics of the AGT-related disease or condition in the subject To decide, (ii) The baseline pre-treatment physiological characteristics of the AGT-related disease or condition, This includes comparing the determined physiological characteristics, From the above comparison, the effectiveness of the double-stranded ribonucleic acid (dsRNA) agent administered to the subjects can be determined. The method according to claim 64, wherein one or more of the following are indicated: and levels.

66. The determined physiological characteristics are essential hypertension, secondary hypertension, and hypertensive emergency (malignant hypertension). (including hypertension and accelerated hypertension), acute hypertension, pregnancy-related hypertension, and treatment-resistant hypertension. The method according to claim 65, wherein hypertension includes high blood pressure.

67. The reduction in the subject's blood pressure indicates the effectiveness of administering the double-stranded ribonucleic acid (dsRNA) agent to the subject. The method according to claim 65, which demonstrates the presence of sex.

68. The level of AGT protein in the subject is determined by the base level of AGT protein in the subject. A method for reducing the level of AGT gene expression compared to the pre-treatment level of Sline To reduce it, double-stranded ribonucleic acid (dsRNA) according to any one of claims 1 to 36. ) an agent, or the composition according to any one of claims 37 to 41, administered to the subject in an effective amount. A method that includes doing something.

69. The method according to claim 68, wherein the dsRNA agent is administered subcutaneously or intravenously to the subject.

70. The physiological characteristics of AGT-related diseases or conditions in the subject are defined as the AGT-related characteristics in the subject. A method for modifying pre-treatment physiological characteristics of a disease or condition compared to baseline, To change the physiological characteristics of AGT-related diseases or conditions in the subject, A double-stranded ribonucleic acid (dsRNA) agent according to any one of claims 1 to 36, or claim 37. A method comprising administering to the subject an effective amount of the composition described in any one of items 41.

71. The method according to claim 70, wherein the dsRNA agent is administered subcutaneously or intravenously to the subject.

72. The aforementioned physiological characteristics include essential hypertension, secondary hypertension, hypertensive emergency (malignant hypertension and This includes acute hypertension (such as accelerated hypertension), pregnancy-related hypertension, and treatment-resistant hypertension. The method according to claim 70, wherein the patient has hypertension.

73. Equation (A): 5'-Z １ AGCUUGUUUGUGAAACZ ２ -3' and 0, 1, 2, also It contains three different sense strands, and in the formula, Z １ However, 0-15 nucleotide motif A nucleotide sequence containing 'f', and Z ２ However, is it selected from one of A, U, C, or G? The dsRNA agent according to claim 1, or one that does not exist.

74. Formula (B): 5'-Z ３ GUUUCACAAAACAAGCUZ ４ -3' and 0, 1, 2, Or the formula contains three different antisense strands, where Z ３ However, A, U, C, G One is selected from or does not exist, and Z ４ However, it contains 0-15 nucleotide motifs A dsRNA agent according to claim 1, wherein the nucleotide sequence is a dsRNA agent.

Images