JPWO2020061177A5 - - Google Patents

Description

Alcohol- related liver disease (ARLD) is also prevalent worldwide and represents a progressive liver disease caused by excessive and sustained alcohol use. Various disease states of ARLD exist and include alcoholic fatty liver (alcoholic fatty liver disease), alcoholic hepatitis, and cirrhosis.

Novel HSD17B13 gene-specific RNA interference (RNAi) agents (also referred to herein as RNAi agents, RNAi triggers, or triggers), such as double-stranded RNAi that can selectively and effectively inhibit the expression of the HSD17B13 gene agent is required. Additionally, there is a need for compositions containing novel HSD17B13-specific RNAi agents to treat diseases such as alcoholic or non-alcoholic liver diseases, including NAFLD, NASH, liver fibrosis, and cirrhosis, among others.

The described HSD17B13 RNAi agents can be used for therapeutic treatment (prophylactic and preventative) of conditions and diseases associated with alcoholic or non-alcoholic liver diseases, including NAFLD, NASH, liver fibrosis, and cirrhosis. ) can be used in methods for (including treatment). The methods disclosed herein involve administering one or more HSD17B13 molecules to a subject, e.g., a human or animal subject, using any suitable method known in the art, such as subcutaneous injection or intravenous administration. and administering an RNAi agent.

In a further aspect, the present disclosure provides methods for treating diseases or conditions caused by alcoholic or non-alcoholic liver diseases, including NAFLD, NASH, liver fibrosis, and/or cirrhosis (prophylactic and prophylactic). (preventative treatment), the method comprises administering to a subject in need thereof an HSD17B13 RNAi agent having an antisense strand comprising any of the sequences in Table 2 or 3. A method comprising: In some embodiments, herein are methods of treating diseases or conditions caused by alcoholic or non-alcoholic liver diseases, including NAFLD, NASH, liver fibrosis, and/or cirrhosis (prophylactic). (preventative) treatment), the method comprising administering to a subject in need thereof an HSD17B13 RNAi agent having a sense strand comprising any of the sequences in Table 2 or 3. , describes the method. Also described herein are compositions for use in such methods.

Pharmaceutical compositions comprising HSD17B13 RNAi agents and methods disclosed herein include administering to a subject a therapeutically effective amount of an HSD17B13 RNAi agent described herein, thereby increasing the expression of HSD17B13 mRNA in the subject. by inhibiting the level of target mRNA in a cell, group of cells, group of cells, tissue, organ, or subject. In some embodiments, the subject has been previously identified or diagnosed as having pathogenic upregulation of the target gene in the target cell or tissue. In some embodiments, the subject has been previously identified or diagnosed with alcoholic or non-alcoholic liver disease, such as NAFLD, NASH, liver fibrosis, and/or cirrhosis. In some embodiments, the subject suffers from a condition associated with alcoholic or non-alcoholic liver disease, such as NAFLD, NASH, liver fibrosis, and/or cirrhosis.

In some embodiments, the described pharmaceutical compositions comprising HSD17B13 RNAi agents are used to treat alcoholic or non-alcoholic liver disease, including NAFLD, NASH, liver fibrosis, cirrhosis, and excess HSD17B13 in a subject. The occurrence and associated clinical symptoms are used for treatment or management. In some embodiments, a therapeutically (including prophylactically) effective amount of one or more pharmaceutical compositions is administered to a subject in need of such treatment. In some embodiments, administration of any of the disclosed HSD17B13 RNAi agents can be used to reduce the number, severity, and/or frequency of symptoms of a disease in a subject.

In some embodiments, the described HSD17B13 RNAi agents may optionally be combined with one or more additional therapeutic agents. The HSD17B13 RNAi agent and additional therapeutic agent may be administered in a single composition or may be administered separately. In some embodiments, one or more therapeutic agents are administered separately from the RNAi agent in a separate dosage form (e.g., the HSD17B13 RNAi agent is administered by subcutaneous injection, but the additional therapeutic agent is not related to the method of therapeutic administration regimen). administered orally). In some embodiments, a described HSD17B13 RNAi agent is administered to a subject in need thereof by subcutaneous injection, and one or more optional additional therapeutic agents are administered orally, both of which are used to treat NAFLD, NASH, Treatment regimens are provided for diseases and conditions associated with alcoholic or non-alcoholic liver disease, including liver fibrosis and/or cirrhosis. In some embodiments, a described HSD17B13 RNAi agent is administered to a subject in need thereof by subcutaneous injection, and one or more optional additional therapeutic agents are administered by a separate subcutaneous injection. In some embodiments, the HSD17B13 RNAi agent and one or more additional therapeutic agents are combined into a single dosage form (eg, a "cocktail" formulated into a single composition for subcutaneous injection). A HSD17B13 RNAi agent, with or without one or more additional therapeutic agents, can be combined with one or more excipients to produce a pharmaceutical composition.

Methods of Treatment and Inhibition of Expression The HSD17B13 RNAi agents disclosed herein are used to treat a subject (e.g., a human or other mammal) with a disease or disorder that would benefit from administration of the RNAi agent. be able to. In some embodiments, the RNAi agents disclosed herein are used to reduce HSD17B13 mRNA expression and/or HSD17B13 (alternatively referred to herein as 17β-HSD13) protein levels and/or Subjects who would benefit from inhibition, such as those diagnosed with or suffering from conditions associated with alcoholic or non-alcoholic liver disease, including NAFLD, NASH, liver fibrosis, and cirrhosis. Can be treated.

In some embodiments, methods of treating diseases, disorders, or conditions caused by alcoholic or non-alcoholic liver disease, including NAFLD, NASH, liver fibrosis, and/or cirrhosis (prophylactic) or preventative treatment), the method comprises administering a therapeutically effective amount of an HSD17B13 RNAi agent comprising an antisense strand that is at least partially complementary to a portion of HSD17B13 mRNA having the sequence of Table 1. , to a subject in need thereof. In some embodiments, methods of treating diseases or conditions caused by alcoholic or non-alcoholic liver disease, including NAFLD, NASH, liver fibrosis, and/or cirrhosis (prophylactic or prophylactic (preventative treatment), the method comprises an antisense strand comprising a sequence of any of the sequences of Table 2 or 3, and an antisense strand that is at least partially complementary to the antisense strand; Disclosed herein is a method comprising administering to a subject in need thereof a therapeutically effective amount of an HSD17B13 RNAi agent comprising a sense strand comprising any of the following sequences. In some embodiments, methods of treating diseases or conditions caused by alcoholic or non-alcoholic liver disease, including NAFLD, NASH, liver fibrosis, and/or cirrhosis (prophylactic or prophylactic a sense strand comprising any of the sequences of Table 2 or 4, and a sequence of Table 2 or 3 that is at least partially complementary to the sense strand. Disclosed herein are methods comprising administering to a subject in need thereof a therapeutically effective amount of an HSD17B13 RNAi agent comprising an antisense strand comprising any of the sequences thereof.

The use of HSD17B13 RNAi agents can be used in the therapeutic (including prophylactic) treatment of diseases/disorders associated with alcoholic or non-alcoholic liver disease, including NAFLD, NASH, liver fibrosis, and cirrhosis; Provided are methods for promoting or increasing HSD17B13 expression. The HSD17B13 RNAi agents described mediate RNA interference inhibiting the expression of one or more genes required for the production of HSD17B13 protein. HSD17B13 RNAi agents can also be used to treat or prevent various diseases, disorders, or conditions, including alcoholic or non-alcoholic liver diseases, including NAFLD, NASH, liver fibrosis, and/or cirrhosis. . Additionally, compositions for in vivo delivery of HSD17B13 RNAi agents to hepatocytes are described.

Claims (27)

An RNAi agent for inhibiting HSD17B13 gene expression,
The RNAi agent is:
an antisense strand comprising at least 17 consecutive nucleotides that differ by 0 or 1 nucleotide from the sequence set forth in SEQ ID NO: 3 ;
a sense strand comprising a nucleotide sequence that is at least partially complementary to the antisense strand;
An RNAi agent comprising: 2. The RNAi agent of claim 1, wherein the RNAi agent is linked to a targeting ligand. 3. The RNAi agent of claim 2, wherein the targeting ligand comprises N -acetyl-galactosamine. The targeting ligands are: (NAG13), (NAG13)s, (NAG18), (NAG18)s, (NAG24), (NAG24)s, (NAG25), (NAG25)s, (NAG26), (NAG26)s , (NAG27), (NAG27)s, (NAG28), (NAG28)s, (NAG29), (NAG29)s, (NAG30), (NAG30)s, (NAG31), (NAG31)s, (NAG32), (NAG32)s, (NAG33), (NAG33)s, (NAG34), (NAG34)s, (NAG35), (NAG35)s, (NAG36), (NAG36)s, (NAG37), (NAG37)s, (NAG38), (NAG38)s, (NAG39), and (NAG39)s
4. The RNAi agent according to claim 3, comprising a structure selected from the group consisting of. 5. The RNAi agent according to claim 4, wherein the targeting ligand comprises a structure of (NAG37) or (NAG37)s . The antisense strand has the following nucleotide sequence (5'→3'):
usCfsasUfcUfaUfcAfgAfcUfuCfuUfaCfsg (SEQ ID NO: 2); or
usCfsasUfcUfaucagAfcUfuCfuUfaCfsg (SEQ ID NO: 4);
comprises , consists of, or consists essentially of a modified nucleotide sequence that differs by zero or one nucleotides from one of
In the above sequence, a, c, g, and u are 2'-O-methyladenosine, 2'-O-methylcytidine, 2'-O-methylguanosine, and 2'-O-methyluridine, respectively. ;Af, Cf , and Uf are 2'-fluoroadenosine, 2'-fluorocytidine , and 2'-fluorouridine, respectively; s is a phosphorothioate bond;
all or substantially all of the nucleotides on the sense strand are modified nucleotides;
The RNAi agent according to any one of claims 1 to 5 . The sense strand has the following nucleotide sequence (5'→3'):
scguaagaaGfUfCfugauagaugas (SEQ ID NO: 14); or
scguaagaaGfuCfuGfauagagas (SEQ ID NO: 15)
comprises, consists of, or consists essentially of a modified nucleotide sequence that differs by zero or one nucleotides from one of
In the above sequence, a, c, g, and u are 2'-O-methyladenosine, 2'-O-methylcytidine, 2'-O-methylguanosine, and 2'-O-methyluridine, respectively. ; Cf, Gf, and Uf are 2'-fluorocytidine, 2'-fluoroguanosine, and 2'-fluorouridine, respectively; s is a phosphorothioate bond;
all or substantially all of the nucleotides on the antisense strand are modified nucleotides;
The RNAi agent according to any one of claims 1 to 6. The RNAi agent according to any one of claims 1 to 7 , wherein the RNAi agent comprises a double-stranded structure of : AD06214 (SEQ ID NO: 2 and 14 ) or AD06280 (SEQ ID NO: 4 and 15). The antisense chain contains a modifiered nucleotide sequence of (5 '→ 3') USCFSASUFCUFCAFGUFUCUFUFUFUFACFSG (Semorable number 2), and the sense chain is (5 '→ 3') (NAG37) S (Invab) SCGUA. AGAAGFUFCFUGAUAGAUGAS (Invab) ( comprising the modified nucleotide sequence of SEQ ID NO: 14);
In the above sequence, a, c, g, and u are 2'-O-methyladenosine, 2'-O-methylcytidine, 2'-O-methylguanosine, and 2'-O-methyluridine, respectively. ; Af, Cf, Gf, and Uf are 2'-fluoroadenosine, 2'-fluorocytidine, 2'-fluoroguanosine, and 2'-fluorouridine, respectively; s is a phosphorothioate bond; (invAb) is an inverted abasic deoxyribose residue; (NAG37)s has the following chemical structure:
The RNAi agent according to claim 1, comprising: The antisense strand comprises a modified nucleotide sequence of (5'→3')usCfsasUfcUfaucagAfcUfuCfuUfaCfsg (SEQ ID NO: 4), and the sense strand comprises a modified nucleotide sequence of (5'→3')(NAG37)s(invAb)scguaagaaGfuCfuGfauagaugas(invAb)( comprising the modified nucleotide sequence of SEQ ID NO: 15);
In the above sequence, a, c, g, and u are 2'-O-methyladenosine, 2'-O-methylcytidine, 2'-O-methylguanosine, and 2'-O-methyluridine, respectively. ; Af, Cf, Gf, and Uf are 2'-fluoroadenosine, 2'-fluorocytidine, 2'-fluoroguanosine, and 2'-fluorouridine, respectively; s is a phosphorothioate bond; (invAb) is an inverted abasic deoxyribose residue; (NAG37)s has the following chemical structure:
The RNAi agent according to claim 1, comprising : The RNAi agent is shown in Figures 1A to 1D below :
Chemical structure described in Figures 2A to 2D below :
Chemical structures described in Figures 6A to 6D below :

or the chemical structure described in Figures 7A to 7D below:
The RNAi agent according to claim 1, comprising the chemical structure according to claim 1. The RNAi agent is shown in Figures 2A to 2D below :
The RNAi agent according to claim 1, comprising the chemical structure according to claim 1. The RNAi agent is shown in Figures 7A to 7D below :

The RNAi agent according to claim 1, comprising the chemical structure according to claim 1. A composition comprising an RNAi agent according to any one of claims 1 to 13 , further comprising a pharmaceutically acceptable excipient. 15. The composition of claim 14 , wherein the composition further comprises a second RNAi agent for inhibiting expression of HSD17B13. 16. The composition of claim 14 or 15 , wherein the composition further comprises one or more additional therapeutic agents. A composition according to any one of claims 14 to 16, which is a pharmaceutical composition. 18. The pharmaceutical composition of claim 17 for the treatment of a disease, disorder, or condition mediated at least in part by HSD17B13 gene expression. 19. The pharmaceutical composition according to claim 18, wherein the disease is alcohol-related liver disease. 19. The pharmaceutical composition according to claim 18, wherein the disease is non-alcoholic fatty liver disease (NAFLD). 21. The pharmaceutical composition according to claim 20, wherein the disease is non-alcoholic steatohepatitis (NASH). A method of inhibiting the expression of the HSD17B13 gene in cells in vitro , the method comprising using the RNAi agent according to any one of claims 1 to 13 or the RNAi agent according to any one of claims 14 to 16 . A method comprising introducing an effective amount of a composition into a cell. 23. The method of claim 22, wherein the cells are derived from a human subject. Use of an RNAi agent according to any one of claims 1 to 13 or a composition according to any one of claims 14 to 16 for the treatment of diseases mediated at least in part by HSD17B13 gene expression, Use for the manufacture of a medicament for the treatment of a disorder or condition. 25. The use according to claim 24, wherein the disease is alcohol-related liver disease. 25. The use according to claim 24 , wherein the disease is non-alcoholic fatty liver disease (NAFLD). 27. The use according to claim 26, wherein the disease is non-alcoholic steatohepatitis (NASH).

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