Inhibitors of expression and / or function

Abstract

This application relates to inhibitors, compositions containing inhibitors, and their use in the treatment or prevention of vascular and / or metabolic diseases.

Description

Technical Field

[0001] This application provides inhibitors suitable for therapeutic use, such as nucleic acid compounds, like siRNA. Furthermore, this application provides methods for preparing these compounds, and methods for using these compounds to treat various diseases and conditions. Background Technology

[0002] Inhibitors, such as oligonucleotides, can suppress gene expression and / or the expression or function of other targets, such as LNCRNAs, and have significant therapeutic applications in medicine. Oligonucleotides can be used to silence genes that cause specific diseases. Gene silencing prevents protein synthesis by inhibiting translation. Importantly, gene silencers are a promising alternative to traditional small-molecule organic compounds that inhibit the function of disease-related proteins. siRNA, antisense RNA, and microRNA are all oligonucleotides that prevent protein synthesis through gene silencing.

[0003] Over the past two decades, numerous modified siRNA compounds have been developed for diagnostic and therapeutic purposes, including siRNA / RNAi therapeutics for treating a wide range of diseases, including central nervous system disorders, inflammatory diseases, metabolic disorders, tumors, infectious diseases, and ophthalmic diseases.

[0004] This application relates to inhibitors such as oligomers (e.g., nucleic acids, such as oligonucleotides / oligonucleotide compounds), and their use in the treatment and / or prevention of diseases.

[0005] Specifically, suitable inhibitors are still needed to help prevent and / or treat diseases such as vascular and / or metabolic disorders. Summary of the Invention

[0006] This application relates to, and particularly to, the following as defined in the claims: In one aspect, this application relates to inhibitors of B4GALT1 expression and / or function, which are used to prevent and / or treat and / or control insulin resistance, and / or elevated blood glucose levels, and / or elevated blood insulin levels, and / or elevated glycated hemoglobin (HbA1c) levels, and / or elevated blood free fatty acid levels, and / or elevated blood fibrinogen levels, and / or elevated blood total cholesterol levels, and / or elevated blood low-density lipoprotein (LDL) cholesterol levels, and / or elevated blood triglyceride levels.

[0007] On the other hand, this application relates to inhibitors of B4GALT1 expression and / or function, which are used to prevent and / or treat and / or control insulin resistance, and / or elevated blood glucose levels, and / or elevated blood insulin levels, and / or elevated blood HbA1c levels, and / or elevated blood free fatty acid levels, and / or elevated blood fibrinogen levels, and / or elevated blood total cholesterol levels, and / or elevated blood LDL cholesterol levels, and / or elevated blood triglyceride levels, and / or diabetes in patients who have or are at risk of developing metabolic and / or vascular diseases.

[0008] In one aspect, this application relates to an inhibitor for the purpose of preventing and / or treating and / or controlling insulin resistance, preferably wherein said inhibitor is capable of improving insulin resistance.

[0009] In one aspect, this application relates to an inhibitor for the purpose of this application, used to prevent and / or treat and / or control elevated blood glucose levels, preferably wherein said inhibitor is capable of reducing elevated blood glucose levels.

[0010] In one aspect, this application relates to an inhibitor for the purpose of preventing and / or treating and / or controlling elevated blood insulin levels, preferably wherein the inhibitor is capable of reducing elevated blood insulin levels.

[0011] In one aspect, this application relates to an inhibitor for the purpose of preventing and / or treating and / or controlling elevated blood HbA1c levels, preferably wherein the inhibitor is capable of reducing elevated blood HbA1c levels.

[0012] In one aspect, this application relates to inhibitors for the purposes of this application, which are used to prevent and / or treat and / or control elevated blood free fatty acid levels, preferably wherein said inhibitor is capable of reducing elevated blood free fatty acid levels.

[0013] In one aspect, this application relates to an inhibitor for the purpose of preventing and / or treating and / or controlling elevated fibrinogen levels, preferably wherein the inhibitor is capable of reducing elevated fibrinogen levels.

[0014] In one aspect, this application relates to an inhibitor for the purpose of preventing and / or treating and / or controlling elevated total cholesterol levels, preferably wherein the inhibitor is capable of reducing elevated total cholesterol levels.

[0015] In one aspect, this application relates to an inhibitor for the purpose of preventing and / or treating and / or controlling elevated blood LDL cholesterol levels, preferably wherein the inhibitor is capable of reducing elevated blood LDL cholesterol levels.

[0016] In one aspect, this application relates to an inhibitor for the purpose of preventing and / or treating and / or controlling elevated blood triglyceride levels, preferably wherein the inhibitor is capable of reducing elevated blood triglyceride levels.

[0017] On the other hand, this application relates to inhibitors of B4GALT1 expression and / or function for the prevention and / or treatment and / or control of vascular disease in patients, wherein the vascular disease is associated with insulin resistance, and / or elevated levels of free fatty acids, and / or elevated levels of fibrinogen, and / or elevated levels of total cholesterol, and / or elevated levels of LDL cholesterol, and / or elevated levels of triglycerides, and / or diabetes.

[0018] In one aspect, this application relates to an inhibitor for the purpose of preventing and / or treating and / or controlling vascular diseases associated with insulin resistance, preferably wherein said inhibitor is capable of improving insulin resistance.

[0019] In one aspect, this application relates to inhibitors for the purposes of this application, which are used to prevent and / or treat and / or control vascular diseases associated with elevated blood free fatty acid levels, preferably wherein said inhibitor is capable of reducing elevated blood free fatty acid levels.

[0020] In one aspect, this application relates to an inhibitor for the purpose of preventing and / or treating and / or controlling vascular diseases associated with elevated fibrinogen levels, preferably wherein the inhibitor is capable of reducing elevated fibrinogen levels.

[0021] In one aspect, this application relates to an inhibitor for the purpose of preventing and / or treating and / or controlling vascular diseases associated with elevated total cholesterol levels, preferably wherein the inhibitor is capable of reducing elevated total cholesterol levels.

[0022] In one aspect, this application relates to an inhibitor for the purpose of preventing and / or treating and / or controlling vascular diseases associated with elevated blood LDL cholesterol levels, preferably wherein the inhibitor is capable of reducing elevated blood LDL cholesterol levels.

[0023] In one aspect, this application relates to an inhibitor for the purpose of preventing and / or treating and / or controlling vascular diseases associated with elevated blood triglyceride levels, preferably wherein the inhibitor is capable of reducing elevated blood triglyceride levels.

[0024] In one aspect, this application relates to an inhibitor for the purpose of preventing and / or treating and / or controlling vascular disease associated with diabetes, preferably wherein the inhibitor is capable of improving diabetes.

[0025] On the other hand, this application relates to inhibitors of B4GALT1 expression and / or function for the prevention and / or treatment and / or control of obesity, and / or weight gain, and / or metabolic syndrome in patients.

[0026] In one aspect, this application relates to an inhibitor for use in accordance with the purpose of this application, wherein the obesity, and / or weight gain, and / or metabolic syndrome is associated with insulin resistance, and / or elevated blood free fatty acid levels, preferably wherein the inhibitor is capable of reducing elevated blood free fatty acid levels.

[0027] In one aspect, this application relates to inhibitors for the purposes of this application, wherein the obesity, and / or weight gain, and / or metabolic syndrome is associated with insulin resistance.

[0028] In one aspect, this application relates to an inhibitor for the purposes of this application, wherein the obesity, and / or weight gain, and / or metabolic syndrome is associated with elevated blood free fatty acid levels, preferably wherein the inhibitor is capable of reducing elevated blood free fatty acid levels.

[0029] In one aspect, this application relates to an inhibitor for the purposes of this application, wherein the obesity, and / or weight gain, and / or metabolic syndrome is associated with elevated total cholesterol levels, and / or elevated LDL cholesterol levels, and / or elevated triglyceride levels, preferably wherein the inhibitor is capable of reducing elevated total cholesterol levels, and / or elevated LDL cholesterol levels, and / or elevated triglyceride levels.

[0030] In one aspect, this application relates to inhibitors for the purposes of this application, wherein the metabolic syndrome is further or independently associated with elevated total cholesterol levels and / or elevated LDL cholesterol levels and / or elevated triglyceride levels, preferably wherein the inhibitor is capable of reducing elevated total cholesterol levels, and / or elevated LDL cholesterol levels, and / or elevated triglyceride levels.

[0031] In one aspect, this application relates to an inhibitor for the purposes of this application, wherein the metabolic syndrome is further or independently associated with elevated total blood cholesterol levels, preferably wherein the inhibitor is capable of reducing elevated total blood cholesterol levels.

[0032] In one aspect, this application relates to an inhibitor for use in accordance with the purpose of this application, wherein the metabolic syndrome is further or independently associated with elevated blood LDL cholesterol levels, preferably wherein the inhibitor is capable of reducing elevated blood LDL cholesterol levels.

[0033] In one aspect, this application relates to an inhibitor for use in accordance with the purpose of this application, wherein the metabolic syndrome is further or independently associated with elevated blood triglyceride levels, preferably wherein the inhibitor is capable of reducing elevated blood triglyceride levels.

[0034] In one respect, this application relates to an inhibitor for use according to any of the preceding claims, wherein the patient is a mammalian patient, preferably a human patient.

[0035] In one aspect, this application relates to an inhibitor for the use described in any of the preceding claims, wherein the inhibitor of B4GALT1 expression and / or function is an siRNA oligomer.

[0036] In one aspect, this application relates to inhibitors for the purposes of this application, said inhibitors being siRNA oligomers conjugated to one or more ligand portions.

[0037] In a further aspect, this application relates to an inhibitor according to the uses of this application, said inhibitor being a siRNA oligomer having a first strand and a second strand, wherein: i) The first strand of the siRNA has a length of 15-30 nucleotides, preferably 19-25 nucleotides, more preferably 23 or 25 nucleotides; even more preferably 23 nucleotides; and / or ii) The second strand of the siRNA has a length of 15-30 nucleotides, preferably 19-25 nucleotides, and more preferably 21 nucleotides.

[0038] Furthermore, this application relates to an inhibitor according to the purpose of this application, wherein the second positive chain further includes one or more abasic nucleosides located in the terminal region of the second chain, and wherein the abasic nucleosides are linked to adjacent nucleosides via reverse internucleotide bonds.

[0039] Furthermore, this application relates to an inhibitor according to the purpose of this application, wherein the second chain comprises: i. Two or more abase-free nucleotides located in the terminal region of the second chain; and / or ii. Two or more abase-free nucleosides located in the 5' or 3' end region of the second strand; and / or iii. Two or more abasic nucleosides located in the 5' or 3' end region of the second chain, wherein the abasic nucleosides are located at the overhangs described herein; and / or iv. Two or more consecutive abasic nucleosides located in the terminal region of the second chain, wherein preferably, one of the abasic nucleosides is a terminal nucleoside; and / or v. Two or more consecutive abasic nucleosides located in the 5' or 3' end region of the second strand, wherein preferably, one of the abasic nucleosides is located in the 5' or 3' end region of the second strand; and / or vi. In the terminal region of the second chain, at least one base-free nucleotide is linked to an adjacent base-containing nucleotide via a reverse internucleotide bond; and / or vii. In the 5' or 3' end region of the second chain, at least one base-free nucleotide is linked to an adjacent base-containing nucleotide via a reverse internucleotide bond; and / or viii. A base-free nucleoside that is the penultimate nucleoside, which is linked to a non-terminal nucleoside (referred to herein as the penultimate nucleoside) via a reverse bond; and / or ix. When the chain is read along the terminal direction, the baseless nucleosides are connected as two terminal nucleosides by 5'-3' bonds; x. When the chain is read along the end containing the terminal nucleoside, the baseless nucleoside is connected by a 3'-5' bond as two terminal nucleosides; xi. As a nucleoside with no base at the two terminal positions, wherein the penultimate nucleoside and the penultimate nucleoside are connected by a reverse bond, and wherein the reverse bond is a 5'-5' reverse bond or a 3'-3' reverse bond; xii. As a nucleoside with no bases at the two terminal positions, wherein the penultimate nucleoside and the penultimate nucleoside are linked by an anti-bond, and wherein... (1) The reverse bond is a 5'-5' reverse bond, and when read along the end direction containing the terminal abasic nucleotide and the penultimate abasic nucleotide, there is a 3'-5' bond between the terminal abasic nucleotide and the penultimate abasic nucleotide; or (2) The reverse bond is a 3'-3' reverse bond, and when read along the end direction containing the terminal abase nucleoside and the penultimate abase nucleoside, there is a 5'-3' bond between the terminal abase nucleoside and the penultimate abase nucleoside.

[0040] Furthermore, this application relates to an inhibitor according to the purpose of this application, wherein the reverse nucleoside internucleotide bond is located in a terminal region remote from the 5' terminal region of the second chain, or in a terminal region remote from the 3' terminal region of the second chain.

[0041] Furthermore, this application relates to inhibitors for the purposes described herein, wherein the reverse nucleoside internucleotide bond is a 3'-3' reverse bond.

[0042] Furthermore, this application relates to inhibitors for the purposes described herein, wherein the reverse nucleoside internucleotide bond is a 5'-5' reverse bond.

[0043] In a further aspect, this application relates to an inhibitor according to the purpose of this application, wherein one or more nucleosides on the first chain and / or the second chain are modified to form a modified nucleoside.

[0044] Furthermore, this application relates to inhibitors for the purposes of this application, wherein the modification is a modification of the 2'-OH group of the ribose, optionally selected from 2'-Me modification or 2'-F modification.

[0045] Furthermore, this application relates to an inhibitor according to the uses of this application, wherein, starting from position 1 of the first chain, the first chain contains a 2'-F modification at any of positions 14, 2, 6, or any combination thereof.

[0046] Furthermore, this application relates to an inhibitor for the purposes of this application, wherein, starting from position 1 of the second chain, the second chain contains a 2'-F modification at positions 7, and / or 9, and / or 11, and / or 13.

[0047] Furthermore, this application relates to inhibitors according to the uses of this application, wherein the first chain and the second chain respectively contain 2'-Me and 2'-F modifications.

[0048] In a further aspect, this application relates to an inhibitor for the purposes of this application, said inhibitor being siRNA, wherein said siRNA comprises at least one thermally unstable modification, starting from position 1 on the first strand, the modification suitably located at one or more positions from position 1 to position 9 on the first strand, and / or located at one or more positions on the second strand that match positions 1-9 on the first strand, wherein said unstable modification is selected from modified unlocking nucleic acids (UNA) and ethylene glycol nucleic acids (GNA), preferably ethylene glycol nucleic acids.

[0049] Furthermore, this application relates to an inhibitor according to the uses of this application, wherein, starting from position 1 on the first strand, the siRNA contains at least one thermally unstable modification at position 7 on the first strand.

[0050] Furthermore, this application relates to an inhibitor for the purposes of this application, said inhibitor being siRNA, wherein, starting from position 1 of the second strand, the siRNA contains three or more 2'-F modifications at positions 7 to 13 of the second strand, for example, four, five, six, or seven 2'-F modifications at positions 7 to 13 of the second strand.

[0051] Furthermore, this application relates to an inhibitor for the purposes of this application, said inhibitor being siRNA, wherein, counting from position 1 of the second strand, the second strand contains at least three 2'-Me modifications at positions 1 to 6, for example, four, five, or six 2'-Me modifications.

[0052] Furthermore, this application relates to an inhibitor for the purposes of this application, said inhibitor being siRNA, wherein the first strand contains at least five consecutive 2'-Me modifications in the 3' end region, preferably including terminal nucleosides in the 3' end region, or nucleosides at least one or two nucleosides away from the terminal nucleosides in the 3' end region.

[0053] Furthermore, this application relates to an inhibitor for the purposes of this application, said inhibitor being siRNA, wherein the first strand contains seven consecutive 2'-Me modifications in the 3' end region, preferably including terminal nucleosides in the 3' end region.

[0054] Furthermore, this application relates to inhibitors for the purposes described herein, wherein the siRNA oligomer further comprises one or more phosphate thioester nucleoside bonds.

[0055] Furthermore, this application relates to inhibitors for the purposes of this application, wherein the one or more thiophosphate nucleoside internucleotide bonds are located between at least three consecutive positions in a 5' proximal terminal region or a 3' proximal terminal region of the second chain, wherein the proximal terminal region is preferably adjacent to the terminal region, and the terminal region contains one or more abase-free nucleosides of the second chain as defined herein.

[0056] Furthermore, this application relates to inhibitors for the purposes of this application, wherein the one or more thiophosphate nucleoside interbonds are located between at least three consecutive positions in the 5' end region and / or 3' end region of the first chain, wherein preferably, the end positions of the 5' end region and / or 3' end region of the first chain are connected to their adjacent positions by thiophosphate nucleoside interbonds.

[0057] In a further aspect, this application relates to an inhibitor according to the uses of this application, wherein the oligomer is siRNA, and the second strand of the siRNA is directly or indirectly conjugated to one or more ligand moieties, wherein the ligand moieties are generally present in the terminal region of the second strand, preferably located in its 3' terminal region.

[0058] Furthermore, this application relates to an inhibitor according to the uses of this application, wherein the ligand portion comprises: i) one or more GalNAc ligands; and / or ii) One or more GalNAc ligand derivatives; and / or iii) One or more GalNAc ligands and / or GalNAc ligand derivatives conjugated to the siRNA via a linker.

[0059] In a further aspect, this application relates to inhibitors according to the uses of this application, wherein the one or more GalNAc ligands and / or GalNAc ligand derivatives are conjugated directly or indirectly to the 5' or 3' end region of the second strand of the siRNA oligomer, preferably conjugated to its 3' end region.

[0060] Furthermore, this application relates to an inhibitor according to the purpose of this application, wherein the ligand portion comprises: .

[0061] Furthermore, this application relates to an inhibitor for the purpose described in this application, having the following structure: ; in: R1 is selected independently from hydrogen, methyl, and ethyl each time it appears; R2 is selected from the following group: hydrogen, hydroxyl group, -OC 1-3 Alkyl group, -C(=O)OC 1-3 Alkyl, halogen, and nitro groups; X1 and X2 are each independently selected from methylene, oxygen, and sulfur when they appear; m is an integer from 1 to 6; n is an integer from 1 to 10; q, r, s, t, and v are each an independent integer between 0 and 4, but must satisfy the following condition: (i) q and r cannot both be 0; and (ii) s, t and v cannot all be 0 at the same time; Z is an oligomer.

[0062] Furthermore, this application relates to an inhibitor for the purpose described in this application, having the following structure: ; in: r and s are each independent integers selected from 1 to 16; and Z is an oligomer.

[0063] Furthermore, this application relates to inhibitors for the uses described herein, which are formulated as pharmaceutical compositions containing excipients and / or carriers.

[0064] On the other hand, this application relates to pharmaceutical compositions comprising the inhibitor described in this application, and pharmaceutically acceptable excipients or carriers.

[0065] On the other hand, this application relates to nucleic acid or pharmaceutical compositions for the prevention or treatment of vascular diseases, such as cardiovascular diseases. Attached Figure Description

[0066] Figure 1a shows an exemplary linear configuration of the conjugate.

[0067] Figure 1b shows an exemplary branching configuration of the conjugate.

[0068] Figure 2-5 The preferred oligomer-connector-ligand construct of this application is shown.

[0069] Figure 6 This refers to the detailed contents of the chemical formulas described in statements 1-101 disclosed herein.

[0070] Figure 7 shows the details of the chemical formulas described in Clauses 1-56 disclosed herein.

[0071] Figure 8 This is a group of active GalNAc-siRNA molecules with EC50 values ​​less than 100 nM. After 10 serial dilutions (starting from 1000 nM) of GalNAc-siRNA targeting mouse B4GALT1 and co-incubation with primary mouse hepatocytes for 48 hours, the dose-response to B4GALT1 mRNA knockdown in primary mouse hepatocytes was measured. Using GraphPad Prism, the EC50 values ​​were determined by fitting the data to a 4-parameter sigmoid dose-response (variable slope) equation. 50 Values. Four active GalNAc-siRNA molecules, namely ETXM619, ETXM624, ETXM628 and ETXM633, were selected for in vivo pharmacological studies.

[0072] Figure 9 This study summarized the effects of multiple administrations of GalNAc-siRNA (ETXM619, ETXM624, ETXM628, and ETXM633, at a dose of 10 mg / kg) to mouse liver tissue on B4GALT1 mRNA knockdown. The y-axis values ​​represent relative mRNA expression levels compared to the untreated group (n=5). Each data point represents relative mRNA expression level, expressed as mean ± standard deviation of n=3 experiments. The red arrows at the top of the graph indicate the date of administration of the test substance.

[0073] Figure 10This study demonstrated the effect of B4GALT1 mRNA knockdown on plasma LDL-c, glucose, and fibrinogen levels. Plasma samples were collected on day 14 after three administrations of ETXM (10 mg / kg, subcutaneous injection) on days 0, 3, and 7. Compared with the untreated group (n=5), the ETXM-treated group (n=12) showed significantly lower levels of LDL-c, glucose, and fibrinogen in normal C57BL / 6 mice. Data presented in this study are mean ± standard deviation.

[0074] Figure 11 The results of 12 weeks of treatment with 3 mg / kg and 10 mg / kg ETXM1201 showed liver function. B4galt1 mRNA expression levels. Compared to negative control animals, liver... B4galt1 mRNA levels were significantly reduced. The sample size for all groups was N=12. Data presented here are mean ± standard deviation. A simple linear model was used for data analysis. A t-test was used to compare treatment groups. Multiple comparison correction was performed on p-values ​​using the FDR method. # indicates the p-value of the negative control group relative to the 3 mg / kg ETXM1201 group, and * indicates the p-value of the negative control group relative to the 10 mg / kg ETXM1201 group.

[0075] Figure 12 This study demonstrates the effect of inhibiting hepatic B4GALT1 on body weight changes during the study. Compared to negative control animals, B4GALT1 inhibitors significantly reduced body weight gain. N=12 across all groups. Data presented here are mean ± standard deviation. A simple linear model was used for data analysis. t-tests were used to compare treatment groups. Multiple comparison corrections for p-values ​​were performed using the FDR method. # indicates the p-value of the negative control group relative to the 3 mg / kg ETXM1201 group, and * indicates the p-value of the negative control group relative to the 10 mg / kg ETXM1201 group.

[0076] Figure 13The effects of hepatic B4GALT1 inhibition on plasma total cholesterol (a), LDL cholesterol (LDL-c) (b), and total triglycerides (c) throughout the time course are shown, along with plasma FPLC profiles at week 12 (d, e). Compared to negative control animals, B4GALT1 inhibition reduced circulating cholesterol, LDL-c, and triglyceride levels, and decreased VLDL and LDL levels in the FPLC profiles. Sample sizes N = 12–15 in all groups. Except for pooled data shown in (d) and (e), all data presented here are mean ± standard deviation. (a)–(c): Data were analyzed using a generalized additive mixed model. The F-test was used to compare treatment groups. Multiple comparisons were corrected using the FDR method. # indicates p-value of negative control group relative to 3 mg / kg ETXM1201 group, and * indicates p-value of negative control group relative to 10 mg / kg ETXM1201 group.

[0077] Figure 14 The effect of hepatic B4GALT1 inhibition on plasma free fatty acids at week 12 is shown. Compared with the negative control group, B4GALT1 inhibition significantly reduced circulating free fatty acid (FFA) levels. N=12 for all groups. Data presented here are mean ± standard deviation. A simple linear model was used to analyze the data. Robust t-tests with standard errors were used to compare treatment groups. The FDR method was used to correct for p-values ​​in multiple comparisons. * indicates the p-value of the negative control group relative to the 10 mg / kg ETXM1201 group.

[0078] Figure 15 The effect of hepatic B4GALT1 inhibition on plasma fibrinogen levels over the entire time course is shown. B4GALT1 inhibition significantly reduced plasma fibrinogen levels compared to negative control animals. N = 12–15 in all groups. Data presented here are mean ± standard deviation. A generalized additive mixed model was used for data analysis. The F-test was used for comparisons between treatment groups. Multiple comparisons were corrected using the FDR method. * indicates the p-value of the negative control group relative to the 10 mg / kg ETXM1201 group.

[0079] Figure 16The effects of hepatic B4GALT1 inhibition on plasma glucose (a), insulin (b), QUICKI index (c), and HbA1c (d) over the entire time course are shown, along with the results of the oral glucose tolerance test (OGTT) (e, f). Compared to negative control animals, B4GALT1 inhibition reduced glucose, insulin, and HbA1c levels and increased the QUICKI index, indicating enhanced insulin sensitivity. Compared to negative control animals, B4GALT1 inhibition reduced glucose levels and the area under the glucose curve (AUC) in the OGTT. N = 12–15 in all groups. Data presented here are mean ± standard deviation. Time course: Data were analyzed using a generalized additive mixed model. The F-test was used to compare treatment groups. Multiple comparison correction was performed using the FDR method. OGTT curves: Data were analyzed using a generalized additive model. The parametric bootstrap method was used to compare treatment groups. Multiple comparison correction was performed using the FDR method. AUC: Data were analyzed using a simple linear model. Robust t-tests with standard errors were used to compare the treatment groups. Multiple comparisons were corrected for p-values ​​using the FDR method. # indicates the p-value of the negative control group relative to the 3 mg / kg ETXM1201 group, and * indicates the p-value of the negative control group relative to the 10 mg / kg ETXM1201 group. Invention Details This application specifically provides inhibitors, such as oligomers (e.g., nucleic acids, such as repressive RNA molecules (which may be referred to as iRNA or siRNA)), and compositions comprising said inhibitors, said inhibitors being capable of affecting the expression of a target, for example by binding to mRNA transcribed from a gene, or by inhibiting the function of nucleic acids such as long non-coding RNAs (hereinafter referred to as "LNCRNAs"). The target may be intracellular, for example, in cells within a subject (e.g., a human). The inhibitors can be used to prevent and / or treat medical conditions related to, for example, the expression of a target gene or the presence / activity of intracellular nucleic acids (e.g., long non-coding RNAs).

[0081] Specifically, this application identifies that inhibitors of post-translational glycosylation (e.g., B4GALT1 inhibitors) can be used to prevent and / or treat vascular and / or metabolic diseases, as detailed below.

[0082] B4GALT1 is β-1,4-galactosyltransferase 1, which is encoded by the B4GALT1 gene (SEQ ID NO:1) in humans.

[0083] definition The “first strand” (also referred to herein as the antisense strand or guide strand, the two terms are used interchangeably) refers to a nucleic acid strand, such as a single strand in siRNA or dsiRNA, that contains a region substantially complementary to the target sequence (e.g., mRNA). The term “complementary region” as used herein refers to a region on the antisense strand that is substantially complementary to the sequence (e.g., the target sequence). When this complementary region is not perfectly complementary to the target sequence, mismatches may occur in internal or terminal regions of the molecule. In some embodiments, double-stranded nucleic acids (e.g., the siRNA reagent of this application) contain nucleotide mismatches in the antisense strand.

[0084] The “second strand” (also referred to as the sense strand or the follower strand in this paper, which are used interchangeably) refers to a nucleic acid strand (e.g., siRNA) that contains a region substantially complementary to the region of the term antisense strand as defined in this paper.

[0085] In the context of a molecule containing a nucleic acid with a ligand-linked portion (optionally also including a linker portion), the nucleic acid described in this application may be referred to as an oligonucleotide portion or oligonucleotide portion.

[0086] Oligonucleotides are short-chain nucleic acid polymers. While oligonucleotides contain phosphodiester bonds between nucleoside components (bases plus sugars), this application is not limited to oligonucleotides where adjacent nucleosides are always linked by such phosphodiester bonds, but also considers other nucleoside oligomers linked by non-phosphate bonds. For example, the bond between nucleosides may be a thiophosphate bond. Therefore, the term "oligonucleotide" herein encompasses both oligonucleotides and other nucleoside oligomers. According to this application, nucleic acid oligonucleotides that are at least partially oligonucleotides are preferred. According to this application, oligonucleotides having one or more or mostly phosphodiester backbone bonds between nucleosides are also preferred. Furthermore, according to this application, oligonucleotides having one or more or mostly phosphodiester backbone bonds between nucleosides, and also having one or more thiophosphate backbone bonds between nucleosides (typically located in the terminal regions of the first and / or second chains) are also preferred.

[0087] In some embodiments, the double-stranded nucleic acid (e.g., siRNA reagent) described in this application contains nucleoside mismatches in its positive strand. In some embodiments, the nucleoside mismatches are, for example, located within 5, 4, 3, 2, or 1 nucleoside from the 3' end of the nucleic acid, such as siRNA.

[0088] In another embodiment, the nucleoside mismatch is located at the 3' end nucleoside of, for example, a nucleic acid (such as siRNA).

[0089] A “target sequence” (also known as target RNA or target mRNA) is a continuous nucleoside sequence within an mRNA molecule formed during gene transcription. It can include mRNA produced by RNA processing of primary transcription products, or it can be a continuous nucleotide sequence in any RNA molecule that needs to be repressed (such as LNCRNA).

[0090] The target sequence length can be approximately 10-35 nucleotides, such as approximately 15-30 nucleotides. Specifically, target sequence lengths can be approximately 15-30, 15-29, 15-28, 15-27, 15-26, 15-25, 15-24, 15-23, 15-22, 15-21, 15-20, 15-19, 15-18, 15-17, 18-30, 18-29, 18- 28, 18-27, 18-26, 18-25, 18-24, 18-23, 18-22, 18-21, 18-20, 19-30, 19-29, 19-28, 19-27, 19-26, 19-25, 19-24, 19-23, 19-22, 19-21, 19-20, 20-30, 20-29, 20-28, 20-27, 20-26, 20-25, 20-24, 20-23, 20-22, 20-21, 21-30, 21-29, 21-28, 21-27, 21-26, 21-25, 21-24, 21-23 or 21-22 nucleosides. Furthermore, other ranges and lengths falling between those described above also fall within the scope of this application.

[0091] The terms “ribonucleoside” or “nucleoside” can also refer to modified nucleosides as further detailed below.

[0092] Nucleic acids can be DNA or RNA and may contain modified nucleosides. RNA is the preferred nucleic acid.

[0093] In this document, the terms “iRNA,” “siRNA,” “RNAi reagent,” “iRNA reagent,” and “RNA interference agent,” which are used interchangeably, refer to agents containing RNA that mediate the targeted cleavage of RNA transcripts via the RNA-induced silencing complex (RISC) pathway. siRNA guides the sequence-specific degradation of mRNA through RNA interference (RNAi).

[0094] In this document, double-stranded RNA is referred to as "double-stranded siRNA (dsiRNA) reagent," "double-stranded siRNA (dsiRNA) molecule," "double-stranded RNA (dsRNA) reagent," "double-stranded RNA (dsRNA) molecule," "dsiRNA reagent," "dsiRNA molecule," or "dsiRNA," referring to a ribonucleic acid molecular complex having a double-stranded structure comprising two antiparallel and substantially complementary nucleic acid strands, which have "sense" and "antisense" orientations relative to the target RNA, respectively. The majority of the nucleosides in each strand of the nucleic acid (e.g., dsRNA molecule) are preferably ribonucleosides; however, in this case, each or both strands may also contain one or more non-ribonucleosides, such as deoxyribonucleosides or modified ribonucleosides. Furthermore, as used in this specification, "siRNA" may contain chemically modified ribonucleosides.

[0095] The term "modified nucleoside" refers to a nucleoside whose glycosyl moiety, internucleotide bond, or nucleobase has been modified, or any combination thereof. Therefore, the term modified nucleoside encompasses substitution, addition, or removal of, for example, functional groups or atoms, at the internucleotide bond, glycosyl moiety, or nucleobase. Any such modification used in siRNA-type molecules in this specification and claims is covered by "iRNA," "RNAi reagent," "siRNA," or "siRNA reagent."

[0096] The length of the double-stranded region of the nucleic acid (e.g., dsRNA) in this application is approximately 9-40 base pairs, such as 9-36 base pairs, for example, approximately 15-30 base pairs, such as approximately 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, or 36 base pairs, such as approximately 15-30, 15-29, 15-28, 15-27, 15-26, 15-25, 15-24, 15-23, 15-22, 15-21, 15-20, 15-19, 15-18, 15-17, 18-30. 18-29, 18-28, 18-27, 18-26, 18-25, 18-24, 18-23, 18-22, 18-21, 18-20, 19-30, 19-29, 19-28, 19-27, 19-26, 19-25, 19-24, 19-23, 19-22, 19-21, 1 9-20, 20-30, 20-29, 20-28, 20-27, 20-26, 20-25, 20-24, 20-23, 20-22, 20-21, 21-30, 21-29, 21-28, 21-27, 21-26, 21-25, 21-24, 21-23 or 21-22 base pairs.

[0097] The two strands that make up a double-stranded structure can be different parts of a larger molecule, or they can be independent molecules such as RNA molecules.

[0098] The term "nucleoside overhang" refers to at least one unpaired nucleoside extending from the double-stranded structure of a double-stranded nucleic acid. The double-stranded nucleic acid may contain at least one nucleoside overhang; or, the overhang may contain at least two, three, four, five, or more nucleosides. The nucleoside overhang may comprise or consist of nucleoside analogs containing deoxynucleosides. The overhang may be located on the sense strand, antisense strand, or any combination thereof. Furthermore, the nucleoside at the overhang may be present at the 5' end, 3' end, or both ends of the antisense or sense strand.

[0099] In some embodiments, the antisense strand has 1-10 nucleosides at the 3' or 5' end, such as 0-3, 1-3, 2-4, 2-5, 4-10, 5-10, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 nucleoside overhangs.

[0100] "Blunt" or "blunt end" refers to a double-stranded nucleic acid that has no unpaired nucleoside at one end, i.e., no nucleoside overhang. The nucleic acids described in this application include those without a nucleoside overhang at one end or without nucleoside overhangs at both ends.

[0101] Unless otherwise stated, when the term "complementary" is used to describe the relationship between a first nucleoside sequence and a second nucleoside sequence, it refers to the ability of an oligonucleotide containing the first nucleoside sequence to hybridize with an oligonucleotide or polynucleotide containing the second nucleoside sequence under certain conditions and form a double-stranded structure, as will be understood by those skilled in the art. For example, these conditions can be stringent conditions, which may include: 400 mM NaCl, 40 mM PIPES pH 6.4, 1 mM EDTA, reaction at 50°C or 70°C for 12–16 hours, followed by washing (see, for example, Molecular Cloning: A Laboratory Manual, Sambrook et al. (1989), Cold Spring Harbor Laboratory Press).

[0102] Complementary sequences in nucleic acids (e.g., dsiRNA) as described herein refer to oligonucleotides or polynucleotides containing a first nucleoside sequence that form base pairs with oligonucleotides or polynucleotides containing a second nucleoside sequence over the entire length of one or both nucleoside sequences. In this document, such sequences may be referred to as “perfectly complementary” to each other. However, when the first sequence is referred to herein as “substantially complementary” to the second sequence, the two sequences may be perfectly complementary, or they may form one or more mismatched base pairs (e.g., 2, 4, or 5 mismatched base pairs, but preferably no more than 5), while retaining their ability to hybridize under the conditions most relevant to their final application (e.g., repressing gene expression via a RISC pathway). When determining complementarity, overhangs should not be considered mismatches. For example, a nucleic acid (e.g., dsRNA) containing one 17-nucleoside oligonucleotide and another 19-nucleoside oligonucleotide, where the longer oligonucleotide contains a 17-nucleoside sequence perfectly complementary to the shorter oligonucleotide, can still be referred to as “perfectly complementary.”

[0103] The “complementary” sequences used in this article may also include, or consist entirely of, non-Watson-Crick base pairs or base pairs formed from non-natural and modified nucleosides, provided that the above requirements regarding their hybridization ability are met. Such non-Watson-Crick base pairs include, but are not limited to, G:U Wobble or Hoogstein base pairs.

[0104] The terms “complementary,” “fully complementary,” and “fundamentally complementary” used in this article can be used for base pairing between the sense and antisense strands of a nucleic acid (e.g., dsiRNA), or between the antisense strand and the target sequence of a double-stranded nucleic acid (e.g., siRNA) reagent.

[0105] In this application, the second strand of the nucleic acid, particularly the dsiRNA for inhibiting B4GALT1 expression, is at least partially complementary to the first strand of the nucleic acid. In some embodiments, the first and second strands of the nucleic acid are partially complementary if the first and second strands form a double-stranded region of at least 17 base pairs in length, and the double-stranded region contains no more than 1, 2, 3, 4, or 5 mismatched base pairs.

[0106] In some embodiments, if the first and second strands of the nucleic acid according to this application form a double-stranded region of at least 19 base pairs in length, and the double-stranded region contains no more than 1, 2, 3, 4, or 5 mismatched base pairs, then the first and second strands of the nucleic acid are partially complementary. In some embodiments, if the first and second strands of the nucleic acid according to this application form a double-stranded region of at least 21 base pairs in length, and the double-stranded region contains no more than 1, 2, 3, 4, or 5 mismatched base pairs, then the first and second strands of the nucleic acid are partially complementary.

[0107] Alternatively, if the first and second strands of the nucleic acid described in this application form a double-stranded region of at least 17 base pairs in length, wherein at least 14, 15, 16, or 17 are complementary base pairs (specifically Watson-Crick base pairs), then the first and second strands of the nucleic acid are partially complementary.

[0108] In some embodiments, if the first and second strands of the nucleic acid described in this application form a double-stranded region of 19 base pairs in length, wherein at least 14, 15, 16, 17, 18, or all 19 base pairs are complementary (specifically, Watson-Crick base pairs), then the first and second strands of the nucleic acid are partially complementary. In some embodiments, if the first and second strands of the nucleic acid described in this application form a double-stranded region of 21 base pairs in length, wherein at least 16, 17, 18, 19, 20, or all 21 base pairs are complementary (specifically, Watson-Crick base pairs), then the first and second strands of the nucleic acid are partially complementary.

[0109] As used herein, a nucleic acid that is at least partially “substantially complementary” or “partially complementary” to messenger RNA (mRNA) refers to a polynucleotide that is substantially or partially complementary to a contiguous segment of the target mRNA (such as the mRNA encoding a gene). In some embodiments, the contiguous segment of the mRNA is a sequence listed in Table 1, namely any one of SEQ ID NO:2-21 or SEQ ID NO:102-201. For example, if the sequence of the polynucleotide is substantially or partially complementary to an uninterrupted portion of the mRNA encoding the target gene, then the polynucleotide is complementary to at least a portion of the mRNA of the target gene.

[0110] Therefore, in some preferred embodiments, the antisense oligonucleotides disclosed herein are completely complementary to the target mRNA sequence.

[0111] In other embodiments, the antisense oligonucleotides disclosed herein are substantially or partially complementary to the target RNA sequence and comprise a continuous nucleoside sequence that is at least about 80% complementary to the equivalent region of the target RNA sequence over its entire length, for example, at least about 85%, 86%, 87%, 88%, 89%, about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% complementary or 100% complementary.

[0112] In some embodiments, the first (antisense) strand of the nucleic acid according to this application is partially or completely complementary to a consecutive segment of RNA transcribed from the B4GALT1 gene. In some embodiments, the first strand of the nucleic acid according to this application is partially or completely complementary to a consecutive segment of at least 17 nucleotides of the B4GALT1 mRNA. In some embodiments, the first strand of the nucleic acid according to this application is partially or completely complementary to a consecutive segment of 17, 18, 19, 20, 21, 22, or 23 nucleotides of the B4GALT1 mRNA. In some embodiments, the first strand of the nucleic acid according to this application is partially or completely complementary to a consecutive segment of 17, 18, or 19 nucleotides of any sequence listed in Table 1 (i.e., any sequence in SEQ ID NO:2-21 or SEQ ID NO:102-201). In some embodiments, the first strand of the nucleic acid described in this application is partially or completely complementary to a continuous segment of 19, 20, 21, 22 or 23 nucleotides of any sequence in SEQ ID NO:102-201.

[0113] In some embodiments, if the first (antisense) strand of the nucleic acid according to this application comprises a continuous nucleoside sequence of at least 17 nucleotides, wherein at least 14, 15, 16, or 17 nucleotides of the continuous nucleoside sequence are complementary to a continuous segment of B4GALT1 mRNA, then the first (antisense) strand is partially complementary to a continuous segment of B4GALT1 mRNA. In some embodiments, the first strand of the nucleic acid according to this application comprises a continuous nucleoside sequence of at least 17 nucleotides, wherein at least 14, 15, 16, or 17 nucleotides of the continuous nucleoside sequence are complementary to a continuous segment of any of the sequences listed in Table 1 (i.e., any sequence in SEQ ID NO:2-21 or SEQ ID NO:102-201). In some embodiments, the first strand of the nucleic acid according to this application comprises a continuous nucleoside sequence of 19 nucleotides, wherein at least 14, 15, 16, 17, 18, or all 19 nucleotides of the continuous nucleoside sequence are complementary to consecutive segments of any sequence listed in Table 1 (i.e., any sequence in SEQ ID NO: 2-21 or SEQ ID NO: 102-201). In some embodiments, the first strand of the nucleic acid according to this application comprises a continuous nucleoside sequence of 21 nucleotides, wherein at least 16, 17, 18, 19, 20, or all 21 nucleotides of the continuous nucleoside sequence are complementary to consecutive segments of any sequence listed in Table 1 (i.e., any sequence in SEQ ID NO: 102-201). In some embodiments, the first strand of the nucleic acid according to this application comprises a continuous nucleoside sequence of 23 nucleosides, wherein at least 18, 19, 20, 21, 22 or all 23 nucleosides of the continuous nucleoside sequence are complementary to a continuous segment of any sequence listed in Table 1 (i.e. any sequence in SEQ ID NO:102-201).

[0114] In some embodiments, the nucleic acid (such as siRNA) described in this application comprises a sense strand that is substantially or partially complementary to an antisense polynucleotide, which in turn is complementary to a target mRNA sequence and comprises a continuous nucleoside sequence, wherein the continuous nucleoside sequence is typically at least about 80% complementary to the equivalent region of the nucleoside sequence of the antisense strand over its entire length, for example, about 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%, or 100% complementary.

[0115] In some embodiments, the nucleic acid (such as siRNA) described in this application includes an antisense strand that is substantially or partially complementary to a target sequence, and the antisense strand comprises a continuous nucleoside sequence that is at least 80% complementary to the target sequence over its entire length, for example, about 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%, or 100% complementary.

[0116] As used herein, when the target mRNA sequence is sufficiently complementary to the nucleic acid (e.g., iRNA) reagent to facilitate target knockdown, "subject" refers to an animal that endogenously or heterologously expresses the target gene, such as mammals, including primates (e.g., humans, non-human primates (e.g., monkeys and chimpanzees)) or non-primates, or birds. In some preferred embodiments, the subject is a human.

[0117] The term "treatment" refers to a beneficial or anticipated outcome, including but not limited to the relief or improvement of one or more symptoms associated with gene expression. "Treatment" can also refer to prolonged survival compared to expected survival without treatment. Treatment may include prevention of complications, such as reducing liver damage in subjects with liver infection.

[0118] The term “management” as used in this article follows its conventional meaning, referring to bringing the symptoms of a subject’s disease to at least control (i.e., controlling the severity of symptoms within a predetermined level), and in some cases, alleviating symptoms without eliminating the underlying cause.

[0119] As used herein, the term "prevent" is defined as eliminating or reducing the likelihood of the occurrence of symptoms of one or more diseases or conditions. For example, the inhibitors disclosed herein may be used to prevent the occurrence of metabolic and / or vascular diseases.

[0120] The term “therapeuticly effective amount” as used in this article is intended to include an amount of nucleic acid (e.g., iRNA) that is sufficient to achieve effective treatment of the disease when administered to a patient to treat a subject with a disease (e.g., by reducing, improving, or maintaining the existing disease or symptoms of one or more diseases or their associated complications).

[0121] As used herein, the term "pharmaceutically acceptable" means a compound, material, composition, or dosage form that is suitable for tissue contact with human and animal subjects without excessive toxicity, irritation, allergic reactions, or other problems or complications, and has a reasonable benefit / risk ratio.

[0122] As used herein, the term "pharmaceutically acceptable carrier" refers to a pharmaceutically acceptable material, composition, or drug carrier, such as a liquid or solid filler, diluent, excipient, manufacturing aid, or solvent encapsulating material, relating to the transport or delivery of a target compound from one organ or body site to another. Each carrier must be "acceptable," meaning it is compatible with other components in the formulation and will not cause harm to the subject receiving treatment.

[0123] When listing the values ​​or ranges of parameters, the intermediate values ​​and ranges listed are also intended to be part of this application.

[0124] The articles “a” and “an” used in this article refer to one or more (i.e., at least one) of the grammatical objects of the article.

[0125] The term “including” as used in this article means the phrase “including but not limited to” and is used interchangeably with “including but not limited to”.

[0126] Unless the context clearly indicates otherwise, the term “or” as used herein is used to mean “and / or” and is used interchangeably with the term “and / or”. For example, “justice chain or antisense chain” should be understood as “justice chain or antisense chain or justice chain and antisense chain”.

[0127] As used herein, the term “about” refers to the standard tolerance range in this field. For example, “about” can be understood as approximately 2 standard deviations from the mean. In some implementations, “about” means +10%. In some implementations, “about” means +5%. When “about” appears before a series of numbers or ranges, it should be understood that “about” may modify each number in the series or range.

[0128] The term "at least" preceding a number or series of numbers should be understood to include the number adjacent to the term "at least," as well as all subsequent numbers or integers that can be logically understood from the context. For example, the number of nucleotides in a nucleic acid molecule must be an integer. For example, "at least 18 nucleotides in a nucleic acid molecule of 21 nucleotides" means that 18, 19, 20, or 21 nucleotides have the indicated property. When "at least" appears before a series of numbers or a range, it should be understood that "at least" can modify each number in that series or range.

[0129] As used herein, “not more than” or “less than” should be understood as the value immediately preceding the phrase, and as a lower value or integer up to 0 that can be logically inferred from the context. For example, a double strand with “not more than 2 nucleotides” overhangs can have an overhang length of 2, 1, or 0 nucleotides. When “not more than” appears before a series of numbers or ranges, it should be understood that “not more than” can modify each number in the series or range.

[0130] The terminal region of a chain refers to the last 5 nucleotides, counting from the 5' end or 3' end.

[0131] The nucleobase sequence is the sequence of bases in nucleic acids within an oligomer.

[0132] Those skilled in the art can combine various embodiments of this application as needed.

[0133] target The inhibitory targets disclosed in this article may be, but are not limited to, mRNA, LNCRNA, peptides, proteins, or genes.

[0134] The targets described in this article are involved in the post-translational glycosylation pathway. Inhibition of these targets is preferably beneficial for the prevention or treatment of diabetes. The preferred inhibitory target is B4GALT1, which can be inhibited by suppressing its mRNA or protein expression or function, or both.

[0135] On one hand, the target is the mRNA or long non-coding RNA (LNCRNA) expressed by the gene.

[0136] In a preferred embodiment, the target is the mRNA produced by the expression of the B4GALT1 gene. Table 1 below lists exemplary target sequences on the B4GALT1 mRNA.

[0137] Table 1

[0138] It should be understood that SEQ ID NO:2-21 and SEQ ID NO:102-201 are associated with human (Homo sapiens) mRNA sequences.

[0139] Diseases / symptoms This application relates to inhibitors of B4GALT1 expression and / or function in patients with insulin resistance, and / or elevated blood glucose levels, and / or elevated blood insulin levels, and / or elevated glycated hemoglobin (HbA1c) levels, and / or elevated blood free fatty acid levels, and / or elevated blood fibrinogen levels, and / or elevated blood total cholesterol levels, and / or elevated blood LDL cholesterol levels, and / or elevated blood triglyceride levels. In some embodiments, the patients are those who have or are at risk of developing metabolic or vascular diseases.

[0140] Therefore, certain embodiments of this application relate to inhibitors of B4GALT1 expression and / or function for the prevention and / or treatment and / or control of insulin resistance in patients, preferably, said inhibitor is capable of improving insulin resistance.

[0141] As used in this article, the term "insulin resistance" refers to a state in which cells become less responsive to insulin. Therefore, pancreatic beta cells typically compensate by increasing insulin production and secreting more insulin into the bloodstream to lower blood glucose levels and compensate for insulin resistance. Normally, insulin resistance leads to elevated blood glucose levels.

[0142] Various methods are known in the art for assessing a patient's insulin resistance and / or sensitivity. One commonly used method is the Quantitative Insulin Sensitivity Test Index (QUICKI). The QUICKI score can be calculated using the following formula: QUICKI = 1 / (log(fasting blood glucose, mg / dL) + log(fasting insulin, μU / ml)). Fasting blood glucose and insulin levels can be measured as described herein.

[0143] Surprisingly, this study shows that, compared with mice that did not receive siRNA, mice treated with siRNA that inhibited B4GALT1 expression significantly improved insulin sensitivity in mice fed a high-calorie diet, specifically as evidenced by increased QUICKI scores in these mice (see Example 9 and...). Figure 16 C).

[0144] Insulin resistance in humans is typically defined as a QUICKI score ≤ 0.35. Therefore, a QUICKI score ≤ 0.45 (preferably ≤ 0.4, more preferably ≤ 0.35) indicates insulin resistance or a risk of developing insulin resistance.

[0145] In some embodiments, if a patient's QUICKI score improves after receiving treatment with the inhibitor described in this application, the inhibitor can be deemed to improve the patient's insulin sensitivity and / or reduce insulin resistance. For example, during appropriate treatment, the QUICKI score may improve by at least 1%, at least 2%, at least 3%, at least 4%, at least 5%, at least 6%, at least 7%, at least 8%, at least 9%, at least 10%, at least 11%, at least 12%, at least 13%, at least 14%, or at least 15%.

[0146] Another indicator of insulin sensitivity / resistance is the concentration of HbA1c in the blood. HbA1c can potentially reflect average blood glucose levels over the past few weeks or months and can serve as a marker of insulin sensitivity / resistance. HbA1c levels can be used as a diagnostic tool for early detection of insulin sensitivity / resistance, with high HbA1c levels indicating insulin resistance and low HbA1c levels indicating insulin sensitivity.

[0147] This study unexpectedly found that when mice were treated with siRNA that inhibited B4GALT1 expression, the HbA1c levels in mice fed a high-calorie diet were reduced (see Example 9 and...). Figure 16 D).

[0148] In healthy human subjects, HbA1c levels are typically below 6% (42 mmol / mol). Therefore, when a human patient's blood HbA1c level reaches 6% (0.42 mmol / mol) or higher, preferably 6.5% (48 mmol / mol) or higher, the human patient can be described as having insulin resistance or being at risk of developing insulin resistance.

[0149] In some implementations, if a patient's blood HbA1c level decreases after receiving treatment with the inhibitor described in this application, the inhibitor can be deemed to improve the patient's insulin sensitivity and / or reduce insulin resistance. For example, during appropriate treatment, the patient's blood HbA1c level may decrease by at least 1%, at least 2%, at least 3%, at least 4%, at least 5%, at least 6%, at least 7%, at least 8%, at least 9%, or at least 10%.

[0150] To determine a patient's HbA1c level, blood can be collected according to the method described herein, preferably after a fast of at least 8 hours. Subsequently, HbA1c in the blood can be measured using a clinically validated HbA1c test kit, following the manufacturer's instructions.

[0151] Another method for assessing a patient's insulin sensitivity / resistance is the oral glucose tolerance test (OGTT). An OGTT is a medical test that involves orally administering glucose and then collecting a blood sample to determine the rate at which glucose is cleared from the bloodstream. Therefore, an increase in blood glucose and / or insulin concentrations after a glucose load indicates insulin resistance, while a decrease in blood glucose and / or insulin concentrations indicates insulin sensitivity.

[0152] This study unexpectedly found that, compared with control mice that did not receive siRNA treatment, mice fed a high-calorie diet and administered siRNA that inhibited B4GALT1 expression had lower blood glucose levels following a glucose load (see Example 9 and...). Figure 16E and 16F).

[0153] In an oral glucose tolerance test (75 g glucose ingested after a 12-hour fast), insulin resistance may be characterized by insulin levels >100 mIU / L after 60 minutes and >75 mIU / L after 120 minutes. Additionally, or concurrently, insulin resistance in an oral glucose tolerance test (75 g glucose ingested after a 12-hour fast) may also be characterized by blood glucose levels >180 mg / dL (10 mmol / L) after 60 minutes and >140 mg / dL (7.8 mmol / L) after 120 minutes. Therefore, if a patient has an insulin level >100 mIU / L after 60 minutes and >75 mIU / L after 120 minutes in an oral glucose tolerance test (after fasting for 12 hours and then ingesting 75 g of glucose), and / or a glucose level >180 mg / dL (10 mmol / L) after 60 minutes and >140 mg / dL (7.8 mmol / L) after 120 minutes, the patient can be considered to have insulin resistance or be at risk of developing insulin resistance.

[0154] In some embodiments, when a patient’s efficiency in clearing glucose from the blood is improved after receiving treatment with an inhibitor according to the present application, the inhibitor is determined to improve the patient’s insulin sensitivity and / or reduce insulin resistance; preferably, the improvement in blood glucose clearance efficiency is verified by an improved OGTT score.

[0155] An oral glucose tolerance test (OGTT) can be performed by administering an oral glucose load after fasting for at least 8 hours. In humans, the oral glucose dose can be standardized to 75 g dissolved in 300 mL of water. Blood glucose and / or insulin can be measured at t=0 minutes (immediately before glucose administration) and at t=60 and 120 minutes after glucose administration, as described herein.

[0156] In a specific implementation, this application relates to an inhibitor of B4GALT1 expression and / or function for the prevention and / or treatment and / or control of insulin resistance in patients who have or are at risk of developing metabolic and / or vascular diseases, such as any metabolic and / or vascular diseases disclosed herein.

[0157] In a specific implementation, this application relates to a method for preventing and / or treating and / or controlling insulin resistance in a patient, the method comprising administering an inhibitor of B4GALT1 expression and / or function to the patient.

[0158] In a specific implementation, this application relates to a method for preventing and / or treating and / or controlling insulin resistance in patients who have or are at risk of developing metabolic and / or vascular diseases, the method comprising administering an inhibitor of B4GALT1 expression and / or function to the patient.

[0159] In some embodiments, this application relates to inhibitors of B4GALT1 expression and / or function for the prevention and / or treatment and / or control of elevated blood glucose levels in patients.

[0160] This study unexpectedly found that treating mice fed a high-calorie diet with siRNA that inhibits B4GALT1 expression significantly reduced their blood glucose levels (see Example 9 and...). Figure 16 A).

[0161] In healthy individuals, fasting blood glucose levels typically range from 3.9 to 5.6 mmol / L. Therefore, elevated blood glucose levels can be defined as fasting blood glucose levels of at least 5.7 mmol / L, at least 5.8 mmol / L, at least 5.9 mmol / L, at least 6.0 mmol / L, at least 6.1 mmol / L, at least 6.2 mmol / L, at least 6.3 mmol / L, at least 6.4 mmol / L, at least 6.5 mmol / L, at least 6.6 mmol / L, at least 6.7 mmol / L, at least 6.8 mmol / L, at least 6.9 mmol / L, or at least 7.0 mmol / L.

[0162] In some embodiments, the inhibitor is determined to be useful for regulating and / or reducing plasma glucose concentration when a patient's plasma glucose concentration decreases after treatment with the inhibitor according to this application. For example, during appropriate treatment, plasma glucose concentration may be reduced by at least 1%, at least 2%, at least 3%, at least 4%, at least 5%, at least 6%, at least 7%, at least 8%, at least 9%, at least 10%, at least 11%, at least 12%, at least 13%, at least 14%, or at least 15%. Plasma glucose concentration is preferably measured under standard conditions (i.e., fasting). More preferably, plasma glucose concentration is measured after fasting for at least 8 hours.

[0163] In specific implementations, this application relates to an inhibitor of B4GALT1 expression and / or function for the prevention and / or treatment and / or control of elevated blood glucose levels in patients who have or are at risk of developing metabolic and / or vascular diseases (such as any metabolic and / or vascular diseases disclosed herein).

[0164] In a specific implementation, this application relates to a method for preventing and / or treating and / or controlling elevated blood glucose levels in a patient, the method comprising administering an inhibitor of B4GALT1 expression and / or function to the patient.

[0165] In a specific implementation, this application relates to a method for preventing and / or treating and / or controlling elevated blood glucose levels in patients who have or are at risk of developing metabolic and / or vascular diseases, the method comprising administering an inhibitor of B4GALT1 expression and / or function to the patient.

[0166] In a specific implementation, this application relates to inhibitors of B4GALT1 expression and / or function for the prevention and / or treatment and / or control of hyperglycemia. Hyperglycemia is a condition characterized by excessively high levels of glucose in the bloodstream, possibly featuring a fasting blood glucose level higher than 125 mg / dL (6.9 mmol / L).

[0167] In some embodiments, this application relates to inhibitors of B4GALT1 expression and / or function for the prevention and / or treatment and / or control of elevated blood insulin levels in patients.

[0168] This study unexpectedly found that treatment with siRNA that inhibits B4GALT1 expression reduced serum insulin levels in mice fed a high-calorie diet (see Example 9 and...). Figure 16 B).

[0169] In healthy individuals, fasting plasma insulin levels typically range from 5 to 15 μIU / L. Therefore, elevated insulin levels can be defined as fasting plasma insulin levels of at least 16 μIU / L, at least 17 μIU / L, at least 18 μIU / L, at least 19 μIU / L, at least 20 μIU / L, at least 21 μIU / L, at least 22 μIU / L, at least 23 μIU / L, at least 24 μIU / L, at least 25 μIU / L, at least 26 μIU / L, at least 27 μIU / L, at least 28 μIU / L, at least 29 μIU / L, or at least 30 μIU / L.

[0170] In some embodiments, when a patient's plasma insulin concentration decreases after receiving treatment with an inhibitor according to this application, it can be determined that the inhibitor can regulate and / or reduce plasma insulin concentration. For example, during appropriate treatment, plasma insulin concentration may be reduced by at least 1%, at least 2%, at least 3%, at least 4%, at least 5%, at least 6%, at least 7%, at least 8%, at least 9%, at least 10%, at least 11%, at least 12%, at least 13%, at least 14%, or at least 15%. Preferably, plasma insulin concentration is measured under standardized conditions, i.e., in a fasting state. More preferably, plasma insulin concentration is measured after fasting for at least 8 hours.

[0171] In a specific implementation, this application relates to an inhibitor of B4GALT1 expression and / or function, which is used to prevent and / or treat and / or control elevated blood insulin levels in patients who have or are at risk of developing metabolic and / or vascular diseases (such as any metabolic and / or vascular diseases disclosed herein).

[0172] In a specific implementation, this application relates to a method for preventing and / or treating and / or controlling elevated blood insulin levels in a patient, the method comprising administering an inhibitor of B4GALT1 expression and / or function to the patient.

[0173] In a specific implementation, this application relates to a method for preventing and / or treating and / or controlling elevated blood insulin levels in patients who have or are at risk of developing metabolic and / or vascular diseases, the method comprising administering an inhibitor of B4GALT1 expression and / or function to the patient.

[0174] In a specific implementation plan, this application relates to inhibitors of B4GALT1 expression and / or function for the prevention and / or treatment and / or control of hyperinsulinemia. Hyperinsulinemia is a state of excessive circulating insulin in the blood, which may be characterized by fasting plasma insulin levels higher than 20 μIU / mL.

[0175] Those skilled in the art are familiar with methods for determining blood glucose and insulin concentrations. For example, blood can be collected after a fasting period, and plasma can be obtained according to methods known in the art. For humans, the fasting period should last at least 8 hours. Clinically validated kits for determining blood / plasma glucose and insulin concentrations are known to those skilled in the art.

[0176] In some embodiments, this application relates to the use of inhibitors of B4GALT1 expression and / or function in the prevention and / or treatment and / or control of elevated serum HbA1c levels in patients.

[0177] In healthy individuals, HbA1c levels are typically below 6% (42 mmol / mol). Therefore, when HbA1c levels in the blood of human patients reach 6% (0.42 mmol / mol) or higher, preferably 6.5% (48 mmol / mol) or higher, it can be described as an elevated blood HbA1c level.

[0178] Therefore, in some embodiments, if a patient's blood HbA1c level decreases after receiving treatment with the inhibitor of this application, the inhibitor can be deemed to regulate and / or reduce the patient's blood HbA1c level. For example, during an appropriate treatment period, the blood HbA1c level may decrease by at least 1%, at least 2%, at least 3%, at least 4%, at least 5%, at least 6%, at least 7%, at least 8%, at least 9%, or at least 10%.

[0179] In a specific implementation, this application relates to the use of inhibitors of B4GALT1 expression and / or function in the prevention and / or treatment and / or control of elevated serum HbA1c levels in patients who have or are at risk of developing metabolic and / or vascular diseases (such as any of the metabolic and / or vascular diseases disclosed herein).

[0180] In a specific implementation, this application relates to a method for preventing and / or treating and / or controlling elevated blood HbA1c levels in a patient, the method comprising administering an inhibitor of B4GALT1 expression and / or function to the patient.

[0181] In a specific implementation, this application relates to a method for preventing and / or treating and / or controlling elevated blood HbA1c levels in patients who have or are at risk of developing metabolic and / or vascular diseases, the method comprising administering an inhibitor of B4GALT1 expression and / or function to the patient.

[0182] In some embodiments, this application relates to the use of inhibitors of B4GALT1 expression and / or function in the prevention and / or treatment and / or control of elevated levels of free fatty acids in the blood of patients.

[0183] This study unexpectedly found that treating mice fed a high-calorie diet with siRNA that inhibits B4GALT1 expression significantly reduced their circulating free fatty acid levels (see Example 9 and...). Figure 14 Free fatty acids (FFA) are the non-esterified anionic form of fatty acids, mainly derived from the lipolysis of triglycerides. In the blood, FFA mainly circulates in the form of binding with albumin.

[0184] In healthy individuals, the plasma level of circulating free fatty acids is typically in the range of 0.1–0.6 mmol / L. Therefore, an elevated blood level of free fatty acids can be defined as a fasting plasma level of at least 0.6 mmol / L, at least 0.7 mmol / L, at least 0.8 mmol / L, at least 0.9 mmol / L, at least 1 mmol / L, at least 1.1 mmol / L, at least 1.2 mmol / L, at least 1.3 mmol / L, at least 1.4 mmol / L, at least 1.5 mmol / L, at least 1.6 mmol / L, at least 1.7 mmol / L, at least 1.8 mmol / L, at least 1.9 mmol / L, or at least 2 mmol / L.

[0185] In some embodiments, when a patient responds to treatment with the inhibitor according to this application and the concentration of circulating free fatty acids in the patient's plasma decreases, it can be determined that the inhibitor can regulate and / or reduce the concentration of circulating free fatty acids in the plasma. For example, during appropriate treatment, the concentration of circulating free fatty acids in the plasma may decrease by at least 1%, at least 2%, at least 3%, at least 4%, at least 5%, at least 6%, at least 7%, at least 8%, at least 9%, at least 10%, at least 11%, at least 12%, at least 13%, at least 14%, or at least 15%. Preferably, the concentration of circulating free fatty acids in the plasma is measured under standard conditions, i.e., in a fasting state. More preferably, the concentration of circulating free fatty acids in the plasma is measured after fasting for at least 8 hours.

[0186] In a specific implementation, this application relates to the use of inhibitors of B4GALT1 expression and / or function in the prevention and / or treatment and / or control of elevated blood free fatty acid levels in patients who have or are at risk of developing metabolic and / or vascular diseases (such as any of the metabolic and / or vascular diseases disclosed herein).

[0187] In a specific implementation, this application relates to a method for preventing and / or treating and / or controlling elevated levels of free fatty acids in the blood of a patient, the method comprising administering an inhibitor of B4GALT1 expression and / or function to the patient.

[0188] In a specific implementation, this application relates to a method for preventing and / or treating and / or controlling elevated blood free fatty acid levels in patients who have or are at risk of developing metabolic and / or vascular diseases, the method comprising administering an inhibitor of B4GALT1 expression and / or function to the patient.

[0189] Those skilled in the art are familiar with methods for determining the concentration of free fatty acids in blood (particularly plasma). For example, blood can be collected after a fasting period, and plasma can be obtained according to methods known in the art. For humans, the fasting period should last at least 8 hours. Clinically validated kits for determining the concentration of free fatty acids (FFA) in blood / plasma are known to those skilled in the art.

[0190] In some embodiments, this application relates to the use of inhibitors of B4GALT1 expression and / or function in the prevention and / or treatment and / or control of elevated fibrinogen levels in patients.

[0191] This study unexpectedly found that mice fed a high-calorie diet treated with siRNA that inhibits B4GALT1 expression showed a significant reduction in fibrinogen levels in their blood (see Example 9 and...). Figure 15 ).

[0192] In healthy individuals, plasma fibrinogen levels typically range from 200 to 400 mg / dL. Therefore, elevated fibrinogen levels can be defined as fasting plasma fibrinogen levels of at least 400 mg / dL, at least 425 mg / dL, at least 450 mg / dL, at least 475 mg / dL, at least 500 mg / dL, at least 525 mg / dL, at least 550 mg / dL, at least 575 mg / dL, at least 600 mg / dL, at least 625 mg / dL, at least 650 mg / dL, at least 675 mg / dL, at least 700 mg / dL, at least 725 mg / dL, or at least 750 mg / dL.

[0193] In some embodiments, when a patient responds to treatment with the inhibitor according to this application and the concentration of fibrinogen in the patient's plasma decreases, it can be determined that the inhibitor can regulate and / or reduce the concentration of fibrinogen in the plasma. For example, during appropriate treatment, the concentration of fibrinogen in the plasma may decrease by at least 1%, at least 2%, at least 3%, at least 4%, at least 5%, at least 6%, at least 7%, at least 8%, at least 9%, at least 10%, at least 11%, at least 12%, at least 13%, at least 14%, or at least 15%. Preferably, the concentration of fibrinogen in the plasma is measured under standard conditions.

[0194] In a specific implementation, this application relates to the use of inhibitors of B4GALT1 expression and / or function in the prevention and / or treatment and / or control of elevated fibrinogen levels in patients who have or are at risk of developing metabolic and / or vascular diseases (such as any of the metabolic and / or vascular diseases disclosed herein).

[0195] In a specific implementation, this application relates to a method for preventing and / or treating and / or controlling elevated fibrinogen levels in a patient, the method comprising administering an inhibitor of B4GALT1 expression and / or function to the patient.

[0196] In a specific implementation, this application relates to a method for preventing and / or treating and / or controlling elevated fibrinogen levels in patients who have or are at risk of developing metabolic and / or vascular diseases, the method comprising administering an inhibitor of B4GALT1 expression and / or function to the patient.

[0197] Those skilled in the art are familiar with methods for determining fibrinogen levels in blood / plasma. Those skilled in the art are aware of clinically validated kits for determining fibrinogen concentrations in blood / plasma.

[0198] In some embodiments, this application relates to the use of inhibitors of B4GALT1 expression and / or function in the prevention and / or treatment and / or control of elevated total cholesterol levels in patients.

[0199] This study unexpectedly found that treatment with siRNA that inhibits B4GALT1 expression significantly reduced the levels of total cholesterol, LDL cholesterol, and triglycerides in the blood of mice fed a high-calorie diet (see Example 9 and 2010). Figure 14 A-14C). Simultaneously, elevated high-density lipoprotein cholesterol levels were observed in mice receiving this siRNA. Figure 14 D and 14E).

[0200] In healthy individuals, total cholesterol levels are typically below 200 mg / dL. Therefore, elevated blood total cholesterol levels can be defined as fasting plasma total cholesterol levels of at least 200 mg / dL, at least 205 mg / dL, at least 210 mg / dL, at least 215 mg / dL, at least 220 mg / dL, at least 225 mg / dL, at least 230 mg / dL, at least 235 mg / dL, or at least 240 mg / dL.

[0201] In some embodiments, when a patient responds to treatment with the inhibitor according to this application and the patient's plasma total cholesterol concentration decreases, it can be determined that the inhibitor can regulate and / or reduce the concentration of total cholesterol in the plasma. For example, during appropriate treatment, the plasma total cholesterol concentration may decrease by at least 1%, at least 2%, at least 3%, at least 4%, at least 5%, at least 6%, at least 7%, at least 8%, at least 9%, at least 10%, at least 11%, at least 12%, at least 13%, at least 14%, or at least 15%. Preferably, the plasma total cholesterol concentration is measured under standard conditions, i.e., in a fasting state. More preferably, the plasma total cholesterol concentration is measured after fasting for at least 8 hours.

[0202] In a specific implementation, this application relates to the use of inhibitors of B4GALT1 expression and / or function in the prevention and / or treatment and / or control of elevated total cholesterol levels in patients who have or are at risk of developing metabolic and / or vascular diseases (such as any of the metabolic and / or vascular diseases disclosed herein).

[0203] In a specific implementation, this application relates to a method for preventing and / or treating and / or controlling elevated total cholesterol levels in a patient, the method comprising administering an inhibitor of B4GALT1 expression and / or function to the patient.

[0204] In a specific implementation, this application relates to a method for preventing and / or treating and / or controlling elevated total cholesterol levels in patients who have or are at risk of developing metabolic and / or vascular diseases, the method comprising administering an inhibitor of B4GALT1 expression and / or function to the patient.

[0205] In some embodiments, this application relates to the use of inhibitors of B4GALT1 expression and / or function in the prevention and / or treatment and / or control of elevated blood LDL cholesterol levels in patients.

[0206] In healthy individuals, LDL cholesterol levels are typically below 100 mg / dL. Therefore, elevated blood LDL cholesterol levels can be defined as fasting plasma LDL cholesterol levels of at least 100 mg / dL, at least 105 mg / dL, at least 110 mg / dL, at least 115 mg / dL, at least 120 mg / dL, at least 125 mg / dL, at least 130 mg / dL, at least 135 mg / dL, at least 140 mg / dL, at least 145 mg / dL, at least 150 mg / dL, at least 155 mg / dL, or at least 160 mg / dL.

[0207] In some embodiments, when a patient responds to treatment with the inhibitor according to this application and the patient's plasma LDL cholesterol concentration decreases, it can be determined that the inhibitor according to this application can regulate and / or reduce the plasma LDL cholesterol concentration. For example, during appropriate treatment, the plasma LDL cholesterol concentration may decrease by at least 1%, at least 2%, at least 3%, at least 4%, at least 5%, at least 6%, at least 7%, at least 8%, at least 9%, at least 10%, at least 11%, at least 12%, at least 13%, at least 14%, or at least 15%. Preferably, the plasma LDL cholesterol concentration is measured under standard conditions (i.e., fasting). More preferably, the plasma LDL cholesterol concentration is measured after fasting for at least 8 hours.

[0208] In a specific implementation, this application relates to the use of inhibitors of B4GALT1 expression and / or function in the prevention and / or treatment and / or control of elevated blood LDL cholesterol levels in patients who have or are at risk of developing metabolic and / or vascular diseases (such as any of the metabolic and / or vascular diseases disclosed herein).

[0209] In a specific implementation, this application relates to a method for preventing and / or treating and / or controlling elevated blood LDL cholesterol levels in a patient, the method comprising administering an inhibitor of B4GALT1 expression and / or function to the patient.

[0210] In a specific implementation, this application relates to a method for preventing and / or treating and / or controlling elevated blood LDL cholesterol levels in patients who have or are at risk of developing metabolic and / or vascular diseases, the method comprising administering an inhibitor of B4GALT1 expression and / or function to the patient.

[0211] In some embodiments, this application relates to a pharmaceutical composition comprising nucleic acids for treating cardiovascular disease (preferably coronary artery disease), wherein the treatment can lower the level of LDL cholesterol (LDL-c) in the blood.

[0212] In some embodiments, this application relates to a pharmaceutical composition comprising nucleic acids for treating cardiovascular disease (preferably coronary artery disease), wherein the treatment can reduce the level of fibrinogen in the blood.

[0213] In some embodiments, this application relates to the use of inhibitors of B4GALT1 expression and / or function in the prevention and / or treatment and / or control of elevated blood triglyceride levels in patients.

[0214] In healthy individuals, triglyceride levels are typically below 150 mg / dL. Therefore, elevated blood triglyceride levels can be defined as fasting plasma triglyceride levels of at least 150 mg / dL, at least 175 mg / dL, at least 200 mg / dL, at least 225 mg / dL, at least 250 mg / dL, at least 275 mg / dL, at least 300 mg / dL, at least 325 mg / dL, at least 350 mg / dL, at least 375 mg / dL, or at least 400 mg / dL.

[0215] In some embodiments, when a patient responds to treatment with the inhibitor according to this application and the concentration of triglycerides in the patient's plasma decreases, it can be determined that the inhibitor can regulate and / or reduce the concentration of triglycerides in the plasma. For example, during appropriate treatment, the concentration of triglycerides in the plasma may decrease by at least 1%, at least 2%, at least 3%, at least 4%, at least 5%, at least 6%, at least 7%, at least 8%, at least 9%, at least 10%, at least 11%, at least 12%, at least 13%, at least 14%, or at least 15%. Preferably, the concentration of triglycerides in the plasma is measured under standardized conditions (i.e., fasting). More preferably, the concentration of triglycerides in the plasma is measured after fasting for at least 8 hours.

[0216] In a specific implementation, this application relates to the use of inhibitors of B4GALT1 expression and / or function in the prevention and / or treatment and / or control of elevated blood triglyceride levels in patients who have or are at risk of developing metabolic and / or vascular diseases (such as any of the metabolic and / or vascular diseases disclosed herein).

[0217] In a specific implementation, this application relates to a method for preventing and / or treating and / or controlling elevated blood triglyceride levels in a patient, the method comprising administering an inhibitor of B4GALT1 expression and / or function to the patient.

[0218] In a specific implementation, this application relates to a method for preventing and / or treating and / or controlling elevated blood triglyceride levels in patients who have or are at risk of developing metabolic and / or vascular diseases, the method comprising administering an inhibitor of B4GALT1 expression and / or function to the patient.

[0219] Those skilled in the art are familiar with methods for determining cholesterol and triglyceride levels in blood / plasma. Those skilled in the art are aware of clinically validated kits for determining total cholesterol, LDL cholesterol, and / or triglyceride concentrations in blood / plasma.

[0220] In a specific implementation, this application relates to the use of inhibitors of B4GALT1 expression and / or function in the prevention and / or treatment and / or control of insulin resistance, and / or elevated blood glucose levels, and / or elevated blood insulin levels, and / or elevated blood HbA1c levels, and / or elevated blood free fatty acid levels in patients. Preferably, the patient is a patient who has or is at risk of developing a metabolic disease or vascular disease (such as any metabolic disease and / or vascular disease disclosed herein).

[0221] In a specific implementation, this application relates to a method for preventing and / or treating and / or controlling insulin resistance, and / or elevated blood glucose levels, and / or elevated blood insulin levels, and / or elevated blood HbA1c levels, and / or elevated blood free fatty acid levels in a patient, the method comprising administering an inhibitor of B4GALT1 expression and / or function to the patient, preferably, the patient being a patient who has or is at risk of developing metabolic and / or vascular diseases (such as any metabolic and / or vascular diseases disclosed herein).

[0222] In some embodiments, this application relates to the use of inhibitors of B4GALT1 expression and / or function in the prevention and / or treatment and / or control of vascular disease in patients, wherein the vascular disease is associated with insulin resistance, and / or elevated levels of free fatty acids, and / or elevated levels of fibrinogen, and / or elevated levels of total cholesterol, and / or elevated levels of LDL cholesterol, and / or elevated levels of triglycerides, and / or diabetes.

[0223] In some embodiments, this application relates to a method for preventing and / or treating and / or controlling vascular disease in a patient, the method comprising administering to the patient an inhibitor of B4GALT1 expression and / or function, wherein the vascular disease is associated with insulin resistance, and / or elevated levels of free fatty acids, and / or elevated levels of fibrinogen, and / or elevated levels of total cholesterol, and / or elevated levels of LDL cholesterol, and / or elevated levels of triglycerides, and / or diabetes.

[0224] In a specific implementation, this application relates to the use of inhibitors of B4GALT1 expression and / or function in the prevention and / or treatment and / or control of vascular diseases in patients, wherein the vascular diseases are associated with insulin resistance, preferably wherein the inhibitor is capable of improving insulin sensitivity / improving insulin resistance.

[0225] In a specific implementation, this application relates to a method for preventing and / or treating and / or controlling vascular disease in a patient, the method comprising administering to the patient an inhibitor of B4GALT1 expression and / or function, wherein the vascular disease is associated with insulin resistance, preferably wherein the inhibitor is capable of improving insulin sensitivity / improving insulin resistance.

[0226] In other words, in some implementations, patients with or at risk of vascular disease may have the following characteristics: a QUICKI score ≤0.4; and / or a blood HbA1c level ≥6% (0.42 mmol / mol); and / or an insulin level >100 mIU / L after 60 minutes and >75 mIU / L after 120 minutes in an oral glucose tolerance test (75 g glucose ingested after a 12-hour fast); and / or an oral glucose tolerance test (75 g glucose ingested after a 12-hour fast) with a glucose level >180 mg / dL (10 mmol / L) after 60 minutes and >140 mg / dL (7.8 mmol / L) after 120 minutes.

[0227] In some implementations, patients with or at risk of vascular disease may have the following characteristics: a QUICKI score ≤0.35; and / or a blood HbA1c level ≥6.5% (0.48 mmol / mol); and / or an insulin level >100 mIU / L after 60 minutes and >75 mIU / L after 120 minutes in an oral glucose tolerance test (75 g glucose ingested after a 12-hour fast); and / or a glucose level >180 mg / dL (10 mmol / L) after 60 minutes and >140 mg / dL (7.8 mmol / L) after 120 minutes in an oral glucose tolerance test (75 g glucose ingested after a 12-hour fast).

[0228] Preferably, administering an inhibitor of B4GALT1 expression and / or function to patients with or at risk of developing vascular disease can improve insulin sensitivity, thereby preventing, alleviating, or curing the vascular disease. For example, treating patients with insulin resistance and at risk of developing vascular disease using the inhibitor of this application can prevent the occurrence of vascular disease. Similarly, treating patients with insulin resistance and existing vascular disease using the inhibitor of this application can prevent the deterioration or further development of the vascular disease, and may even cure the vascular disease.

[0229] In a specific implementation, this application relates to the use of B4GALT1 expression and / or function inhibitors in the prevention and / or treatment and / or control of vascular diseases in patients, wherein the vascular diseases are associated with elevated blood free fatty acid levels, preferably wherein the inhibitor can reduce elevated blood free fatty acid levels.

[0230] In a specific implementation, this application relates to a method for preventing and / or treating and / or controlling vascular disease in a patient, the method comprising administering a B4GALT1 expression and / or function inhibitor to the patient, wherein the vascular disease is associated with elevated blood free fatty acid levels, preferably wherein the inhibitor can reduce elevated blood free fatty acid levels.

[0231] In other words, in some implementations, patients with or at risk of developing vascular disease may have fasting plasma free fatty acid levels of at least 0.6 mmol / L, at least 0.7 mmol / L, at least 0.8 mmol / L, at least 0.9 mmol / L, at least 1 mmol / L, at least 1.1 mmol / L, at least 1.2 mmol / L, at least 1.3 mmol / L, at least 1.4 mmol / L, at least 1.5 mmol / L, at least 1.6 mmol / L, at least 1.7 mmol / L, at least 1.8 mmol / L, at least 1.9 mmol / L, or at least 2 mmol / L.

[0232] Preferably, administering an inhibitor of B4GALT1 expression and / or function to patients with or at risk of developing vascular disease can reduce the patients' blood free fatty acid levels, thereby preventing, alleviating, or curing vascular disease. For example, treating patients with elevated blood free fatty acid levels and at risk of developing vascular disease using the inhibitor of this application can prevent the occurrence of vascular disease. Similarly, treating patients with elevated blood free fatty acid levels and who already have vascular disease using the inhibitor of this application can prevent the deterioration or further development of vascular disease, and may even cure the vascular disease.

[0233] In a specific implementation, this application relates to the use of B4GALT1 expression and / or function inhibitors in the prevention and / or treatment and / or control of vascular diseases in patients, wherein the vascular diseases are associated with elevated fibrinogen levels, preferably wherein the inhibitor can reduce elevated fibrinogen levels.

[0234] In a specific implementation, this application relates to a method for preventing and / or treating and / or controlling vascular disease in a patient, the method comprising administering a B4GALT1 expression and / or function inhibitor to the patient, wherein the vascular disease is associated with elevated fibrinogen levels, preferably wherein the inhibitor can reduce elevated fibrinogen levels.

[0235] In other words, in some implementations, patients with or at risk of developing vascular disease may have fasting plasma fibrinogen levels of at least 400 mg / dL, at least 425 mg / dL, at least 450 mg / dL, at least 475 mg / dL, at least 500 mg / dL, at least 525 mg / dL, at least 550 mg / dL, at least 575 mg / dL, at least 600 mg / dL, at least 625 mg / dL, at least 650 mg / dL, at least 675 mg / dL, at least 700 mg / dL, at least 725 mg / dL, or at least 750 mg / dL.

[0236] Preferably, administering an inhibitor of B4GALT1 expression and / or function to patients with or at risk of developing vascular disease can reduce the fibrinogen level in these patients, thereby preventing, alleviating, or curing the vascular disease. For example, treating patients with elevated fibrinogen levels and at risk of developing vascular disease using the inhibitor of this application can prevent the occurrence of vascular disease. Similarly, treating patients with elevated fibrinogen levels and existing vascular disease using the inhibitor of this application can prevent the deterioration or further development of the vascular disease, and may even cure the vascular disease.

[0237] In a specific implementation, this application relates to the use of B4GALT1 expression and / or function inhibitors in the prevention and / or treatment and / or control of vascular diseases in patients, wherein the vascular diseases are associated with elevated total cholesterol levels, preferably wherein the inhibitor can reduce elevated total cholesterol levels.

[0238] In a specific implementation, this application relates to a method for preventing and / or treating and / or controlling vascular disease in a patient, the method comprising administering a B4GALT1 expression and / or function inhibitor to the patient, wherein the vascular disease is associated with elevated total blood cholesterol levels, preferably wherein the inhibitor can reduce elevated total blood cholesterol levels.

[0239] In other words, in some implementations, patients with or at risk of developing vascular disease may have fasting plasma total cholesterol levels of at least 200 mg / dL, at least 205 mg / dL, at least 210 mg / dL, at least 215 mg / dL, at least 220 mg / dL, at least 225 mg / dL, at least 230 mg / dL, at least 235 mg / dL, or at least 240 mg / dL.

[0240] Preferably, administering an inhibitor of B4GALT1 expression and / or function to patients with or at risk of developing vascular disease can lower their total cholesterol levels, thereby preventing, alleviating, or curing the vascular disease. For example, treating patients with elevated total cholesterol levels and at risk of developing vascular disease using the inhibitor of this application can prevent the occurrence of vascular disease. Similarly, treating patients with elevated total cholesterol levels and existing vascular disease using the inhibitor of this application can prevent the deterioration or further development of the vascular disease, and may even cure the vascular disease.

[0241] In a specific implementation, this application relates to the use of B4GALT1 expression and / or function inhibitors in the prevention and / or treatment and / or control of vascular diseases in patients, wherein the vascular diseases are associated with elevated blood LDL cholesterol levels, preferably wherein the inhibitor can reduce elevated blood LDL cholesterol levels.

[0242] In a specific implementation, this application relates to a method for preventing and / or treating and / or controlling vascular disease in a patient, the method comprising administering a B4GALT1 expression and / or function inhibitor to the patient, wherein the vascular disease is associated with elevated blood LDL cholesterol levels, preferably wherein the inhibitor can reduce elevated blood LDL cholesterol levels.

[0243] That is, in some implementations, patients who have or are at risk of developing vascular disease may have fasting plasma LDL cholesterol levels of at least 100 mg / dL, at least 105 mg / dL, at least 110 mg / dL, at least 115 mg / dL, at least 120 mg / dL, at least 125 mg / dL, at least 130 mg / dL, at least 135 mg / dL, at least 140 mg / dL, at least 145 mg / dL, at least 150 mg / dL, at least 155 mg / dL, or at least 160 mg / dL.

[0244] Preferably, administering an inhibitor of B4GALT1 expression and / or function to patients with or at risk of developing vascular disease can lower their serum LDL cholesterol levels, thereby preventing, alleviating, or curing vascular disease. For example, treating patients with elevated serum LDL cholesterol levels and at risk of developing vascular disease using the inhibitor of this application can prevent the occurrence of vascular disease. Similarly, treating patients with elevated serum LDL cholesterol levels and existing vascular disease using the inhibitor of this application can prevent the deterioration or further development of vascular disease, and may even cure the vascular disease.

[0245] In a specific implementation, this application relates to the use of B4GALT1 expression and / or function inhibitors in the prevention and / or treatment and / or control of vascular diseases in patients, wherein the vascular diseases are associated with elevated blood triglyceride levels, preferably wherein the inhibitor can reduce elevated blood triglyceride levels.

[0246] In a specific implementation, this application relates to a method for preventing and / or treating and / or controlling vascular disease in a patient, the method comprising administering a B4GALT1 expression and / or function inhibitor to the patient, wherein the vascular disease is associated with elevated blood triglyceride levels, preferably wherein the inhibitor can reduce elevated blood triglyceride levels.

[0247] In other words, in some implementations, patients with or at risk of developing vascular disease may have fasting plasma triglyceride levels of at least 150 mg / dL, at least 175 mg / dL, at least 200 mg / dL, at least 225 mg / dL, at least 250 mg / dL, at least 275 mg / dL, at least 300 mg / dL, at least 325 mg / dL, at least 350 mg / dL, at least 375 mg / dL, or at least 400 mg / dL.

[0248] Preferably, administering an inhibitor of B4GALT1 expression and / or function to patients with or at risk of developing vascular disease can reduce their blood triglyceride levels, thereby preventing, alleviating, or curing the vascular disease. For example, treating patients with elevated blood triglyceride levels and at risk of developing vascular disease using the inhibitor of this application can prevent the occurrence of vascular disease. Similarly, treating patients with elevated blood triglyceride levels and existing vascular disease using the inhibitor of this application can prevent the deterioration or further development of the vascular disease, and may even cure the vascular disease.

[0249] In a specific implementation, this application relates to the use of B4GALT1 expression and / or function inhibitors in the prevention and / or treatment and / or control of vascular diseases in patients, wherein the vascular diseases are associated with diabetes, and preferably, wherein the inhibitor can improve diabetes.

[0250] In a specific implementation, this application relates to a method for preventing and / or treating and / or controlling vascular disease in a patient, the method comprising administering a B4GALT1 expression and / or function inhibitor to the patient, wherein the vascular disease is associated with diabetes, preferably wherein the inhibitor can improve diabetes.

[0251] According to this application, the term "diabetes" as used herein refers to a group of metabolic diseases in which patients have high blood sugar levels due to the body's inability to produce enough insulin or because cells do not respond to the insulin produced. Diabetes is mainly classified into three types: (1) Type 1 diabetes (T1D): arising from the body's inability to produce insulin, currently requiring insulin injections. (Also known as insulin-dependent diabetes, or IDDM, or juvenile diabetes). (2) Type 2 diabetes (T2D): arising from insulin resistance, i.e., cells' inability to effectively utilize insulin, sometimes accompanied by absolute insulin deficiency (formerly known as non-insulin-dependent diabetes, or NIDDM, or adult-onset diabetes). (3) Gestational diabetes (GD): referring to high blood sugar levels in pregnant women with no prior history of diabetes. This condition may indicate subsequent development of type 2 diabetes (T2D). In a preferred embodiment, the diabetes is type 2 diabetes. Without limitation, diabetes can be diagnosed by an oral glucose tolerance test according to the methods disclosed herein.

[0252] Long-term high blood sugar can damage blood vessels and the nerves that control the heart. People with diabetes are also more prone to other conditions that increase the risk of heart disease. For example, high blood pressure increases the pressure of blood flowing through the arteries and can damage the arterial walls. Therefore, people with diabetes are more susceptible to vascular diseases, especially cardiovascular diseases.

[0253] Preferably, administering an inhibitor of B4GALT1 expression and / or function to diabetic patients who have or are at risk of developing vascular disease can control or reverse the patient's diabetes, thereby preventing, alleviating, or curing the vascular disease. For example, treating a patient with diabetes and at risk of developing vascular disease using the inhibitor of this application can prevent the occurrence of vascular disease. Similarly, treating a patient with diabetes and existing vascular disease using the inhibitor of this application can prevent the deterioration or further development of the vascular disease, and may even cure the vascular disease.

[0254] In some embodiments, this application relates to the use of inhibitors of B4GALT1 expression and / or function in the prevention and / or treatment and / or control of obesity, and / or weight gain and / or metabolic syndrome in patients.

[0255] In some embodiments, this application relates to the use of inhibitors of B4GALT1 expression and / or function in the prevention and / or treatment and / or control of obesity, and / or weight gain and / or metabolic syndrome in patients, wherein the obesity and / or weight gain and / or metabolic syndrome is associated with insulin resistance and / or elevated blood free fatty acid levels, preferably, the inhibitor is capable of reducing elevated blood free fatty acid levels.

[0256] In some embodiments, this application relates to methods for preventing and / or treating and / or controlling obesity, and / or weight gain, and / or metabolic syndrome in a patient, said methods comprising administering an inhibitor of B4GALT1 expression and / or function to said patient.

[0257] In some embodiments, this application relates to a method for preventing and / or treating and / or controlling obesity, and / or weight gain, and / or metabolic syndrome in a patient, the method comprising administering to the patient an inhibitor of B4GALT1 expression and / or function, wherein the obesity and / or weight gain and / or metabolic syndrome is associated with insulin resistance, and / or elevated blood free fatty acid levels, preferably, the inhibitor is capable of reducing elevated blood free fatty acid levels.

[0258] In a specific implementation, this application relates to the use of inhibitors of B4GALT1 expression and / or function in the prevention and / or treatment and / or control of obesity and / or weight gain, and / or metabolic syndrome in patients, wherein the obesity and / or weight gain and / or metabolic syndrome is associated with insulin resistance, preferably wherein the inhibitor is capable of improving insulin resistance.

[0259] In a specific implementation, this application relates to a method for preventing and / or treating and / or controlling obesity and / or weight gain and / or metabolic syndrome in a patient, the method comprising administering an inhibitor of B4GALT1 expression and / or function to the patient, wherein the obesity and / or weight gain and / or metabolic syndrome is associated with insulin resistance, preferably wherein the inhibitor is capable of improving insulin resistance.

[0260] In other words, in some implementations, patients with or at risk of obesity and / or weight gain and / or metabolic syndrome may have the following characteristics: a QUICKI score ≤0.4; and / or a blood HbA1c level ≥6% (0.42 mmol / mol); and / or an insulin level >100 mIU / L after 60 minutes and >75 mIU / L after 120 minutes in an oral glucose tolerance test (75 g glucose ingested after a 12-hour fast); and / or an oral glucose tolerance test (75 g glucose ingested after a 12-hour fast) with a glucose level >180 mg / dL (10 mmol / L) after 60 minutes and >140 mg / dL (7.8 mmol / L) after 120 minutes.

[0261] In some implementations, patients with or at risk of obesity and / or weight gain and / or metabolic syndrome may have the following characteristics: a QUICKI score ≤0.35; and / or a blood HbA1c level ≥6.5% (0.48 mmol / mol); and / or an insulin level >100 mIU / L after 60 minutes and >75 mIU / L after 120 minutes in an oral glucose tolerance test (75 g glucose ingested after a 12-hour fast); and / or an oral glucose tolerance test (75 g glucose ingested after a 12-hour fast) with a glucose level >180 mg / dL (10 mmol / L) after 60 minutes and >140 mg / dL (7.8 mmol / L) after 120 minutes.

[0262] Preferably, administering an inhibitor of B4GALT1 expression and / or function to patients who have or are at risk of obesity and / or weight gain can improve insulin sensitivity, thereby preventing or reversing obesity and / or weight gain. For example, treating patients with insulin resistance and at risk of obesity and / or weight gain using the inhibitor of this application can prevent weight gain or induce weight loss. Similarly, treating patients with insulin resistance and who are already obese using the inhibitor of this application can prevent further weight gain or induce weight loss.

[0263] Alternatively, administering an inhibitor of B4GALT1 expression and / or function to patients with or at risk of developing metabolic syndrome can improve their insulin sensitivity, thereby preventing, alleviating, or reversing metabolic syndrome. For example, treating patients with insulin resistance and at risk of developing metabolic syndrome using the inhibitor of this application can prevent the onset of metabolic syndrome in these patients. Similarly, treating patients with insulin resistance and who already have metabolic syndrome using the inhibitor of this application can prevent the worsening or further development of metabolic syndrome, and may even reverse metabolic syndrome.

[0264] In a specific implementation, this application relates to the use of inhibitors of B4GALT1 expression and / or function in the prevention and / or treatment and / or control of obesity and / or weight gain and / or metabolic syndrome in patients, wherein the obesity and / or weight gain and / or metabolic syndrome is associated with elevated blood free fatty acid levels, preferably wherein the inhibitor can reduce elevated blood free fatty acid levels.

[0265] In a specific implementation, this application relates to a method for preventing and / or treating and / or controlling obesity and / or weight gain and / or metabolic syndrome in a patient, the method comprising administering an inhibitor of B4GALT1 expression and / or function to the patient, wherein the obesity and / or weight gain and / or metabolic syndrome is associated with elevated blood free fatty acid levels, preferably wherein the inhibitor can reduce elevated blood free fatty acid levels.

[0266] In other words, in some implementations, patients who have or are at risk of obesity and / or weight gain and / or metabolic syndrome have fasting plasma free fatty acid levels of at least 0.6 mmol / L, at least 0.7 mmol / L, at least 0.8 mmol / L, at least 0.9 mmol / L, at least 1 mmol / L, at least 1.1 mmol / L, at least 1.2 mmol / L, at least 1.3 mmol / L, at least 1.4 mmol / L, at least 1.5 mmol / L, at least 1.6 mmol / L, at least 1.7 mmol / L, at least 1.8 mmol / L, at least 1.9 mmol / L, or at least 2 mmol / L.

[0267] Preferably, administering an inhibitor of B4GALT1 expression and / or function to patients who have or are at risk of obesity and / or weight gain and / or metabolic syndrome can reduce the patients' blood free fatty acid levels, thereby preventing or reversing obesity and / or weight gain. For example, treating patients with elevated blood free fatty acid levels and at risk of obesity and / or weight gain using the inhibitor of this application can prevent weight gain or induce weight loss. Similarly, treating patients with elevated blood free fatty acid levels and who are already obese using the inhibitor of this application can prevent further weight gain or induce weight loss.

[0268] Alternatively, administering an inhibitor of B4GALT1 expression and / or function to patients with or at risk of developing metabolic syndrome can reduce their blood free fatty acid levels, thereby preventing, alleviating, or curing metabolic syndrome. For example, treating patients with elevated blood free fatty acid levels and at risk of developing metabolic syndrome using the inhibitor of this application can prevent the onset of metabolic syndrome in these patients. Similarly, treating patients with elevated blood free fatty acid levels and who already have metabolic syndrome using the inhibitor of this application can prevent the worsening or further development of metabolic syndrome, and may even reverse metabolic syndrome.

[0269] In some embodiments, this application relates to the use of inhibitors of B4GALT1 expression and / or function in the prevention and / or treatment and / or control of obesity and / or weight gain and / or metabolic syndrome in patients, wherein the obesity and / or weight gain and / or metabolic syndrome is associated with elevated total cholesterol levels and / or elevated LDL cholesterol levels and / or elevated triglyceride levels, preferably wherein the inhibitor can reduce elevated total cholesterol levels, and / or elevated LDL cholesterol levels, and / or elevated triglyceride levels.

[0270] In some embodiments, this application relates to methods for preventing and / or treating and / or controlling obesity and / or weight gain and / or metabolic syndrome in a patient, the methods comprising administering an inhibitor of B4GALT1 expression and / or function to the patient, wherein the obesity and / or weight gain and / or metabolic syndrome is associated with elevated total cholesterol levels and / or elevated LDL cholesterol levels and / or elevated triglyceride levels, preferably wherein the inhibitor can reduce elevated total cholesterol levels, and / or elevated LDL cholesterol levels, and / or elevated triglyceride levels.

[0271] In a specific implementation, this application relates to the use of inhibitors of B4GALT1 expression and / or function in the prevention and / or treatment and / or control of obesity and / or weight gain and / or metabolic syndrome in patients, wherein the obesity and / or weight gain and / or metabolic syndrome is associated with elevated total blood cholesterol levels, preferably wherein the inhibitor is capable of reducing elevated total blood cholesterol levels.

[0272] In a specific implementation, this application relates to a method for preventing and / or treating and / or controlling obesity and / or weight gain and / or metabolic syndrome in a patient, the method comprising administering an inhibitor of B4GALT1 expression and / or function to the patient, wherein the obesity and / or weight gain and / or metabolic syndrome is associated with elevated total blood cholesterol levels, preferably wherein the inhibitor is capable of reducing elevated total blood cholesterol levels.

[0273] That is, in some implementations, patients who have or are at risk of obesity and / or weight gain have fasting plasma total cholesterol levels of at least 200 mg / dL, at least 205 mg / dL, at least 210 mg / dL, at least 215 mg / dL, at least 220 mg / dL, at least 225 mg / dL, at least 230 mg / dL, at least 235 mg / dL, or at least 240 mg / dL.

[0274] Preferably, administering an inhibitor of B4GALT1 expression and / or function to patients who have or are at risk of obesity and / or weight gain can reduce their total cholesterol levels, thereby preventing or reversing obesity and / or weight gain. For example, treating patients with elevated total cholesterol levels and at risk of obesity and / or weight gain using the inhibitor of this application can prevent weight gain or induce weight loss. Similarly, treating patients with elevated total cholesterol levels and who are already obese using the inhibitor of this application can prevent further weight gain or induce weight loss.

[0275] In a specific implementation, this application relates to the use of inhibitors of B4GALT1 expression and / or function in the prevention and / or treatment and / or control of obesity and / or weight gain and / or metabolic syndrome in patients, wherein the obesity and / or weight gain and / or metabolic syndrome is associated with elevated blood LDL cholesterol levels, preferably wherein the inhibitor is capable of reducing elevated blood LDL cholesterol levels.

[0276] In a specific implementation, this application relates to a method for preventing and / or treating and / or controlling obesity and / or weight gain and / or metabolic syndrome in a patient, the method comprising administering an inhibitor of B4GALT1 expression and / or function to the patient, wherein the obesity and / or weight gain and / or metabolic syndrome is associated with elevated blood LDL cholesterol levels, preferably wherein the inhibitor is capable of reducing elevated blood LDL cholesterol levels.

[0277] That is, in some implementations, patients who have or are at risk of obesity and / or weight gain have fasting plasma LDL cholesterol levels of at least 100 mg / dL, at least 105 mg / dL, at least 110 mg / dL, at least 115 mg / dL, at least 120 mg / dL, at least 125 mg / dL, at least 130 mg / dL, at least 135 mg / dL, at least 140 mg / dL, at least 145 mg / dL, at least 150 mg / dL, at least 155 mg / dL, or at least 160 mg / dL.

[0278] Preferably, administering an inhibitor of B4GALT1 expression and / or function to patients who have or are at risk of obesity and / or weight gain can reduce their serum LDL cholesterol levels, thereby preventing or reversing obesity and / or weight gain. For example, treating patients with elevated serum LDL cholesterol levels and at risk of obesity and / or weight gain using the inhibitor of this application can prevent weight gain or induce weight loss. Similarly, treating patients with elevated serum LDL cholesterol levels and who are already obese using the inhibitor of this application can prevent further weight gain or induce weight loss.

[0279] In a specific implementation, this application relates to the use of inhibitors of B4GALT1 expression and / or function in the prevention and / or treatment and / or control of obesity and / or weight gain and / or metabolic syndrome in patients, wherein the obesity and / or weight gain and / or metabolic syndrome is associated with elevated blood triglyceride levels, preferably wherein the inhibitor is capable of reducing elevated blood triglyceride levels.

[0280] In a specific implementation, this application relates to a method for preventing and / or treating and / or controlling obesity and / or weight gain and / or metabolic syndrome in a patient, the method comprising administering an inhibitor of B4GALT1 expression and / or function to the patient, wherein the obesity and / or weight gain and / or metabolic syndrome is associated with elevated blood triglyceride levels, preferably wherein the inhibitor is capable of reducing elevated blood triglyceride levels.

[0281] That is, in some implementations, patients who have or are at risk of obesity and / or weight gain have fasting plasma triglyceride levels of at least 150 mg / dL, at least 175 mg / dL, at least 200 mg / dL, at least 225 mg / dL, at least 250 mg / dL, at least 275 mg / dL, at least 300 mg / dL, at least 325 mg / dL, at least 350 mg / dL, at least 375 mg / dL, or at least 400 mg / dL.

[0282] Preferably, administering an inhibitor of B4GALT1 expression and / or function to patients who have or are at risk of obesity and / or weight gain can reduce their blood triglyceride levels, thereby preventing or reversing obesity and / or weight gain. For example, treating patients with elevated blood triglyceride levels and at risk of obesity and / or weight gain using the inhibitor of this application can prevent weight gain or induce weight loss. Similarly, treating patients with elevated blood triglyceride levels and who are already obese using the inhibitor of this application can prevent further weight gain or induce weight loss.

[0283] In some embodiments, this application relates to the use of inhibitors of B4GALT1 expression and / or function in the prevention and / or treatment and / or control of metabolic syndrome in patients, wherein the metabolic syndrome is further or independently associated with elevated total cholesterol levels and / or elevated LDL cholesterol levels and / or elevated triglyceride levels, preferably wherein the inhibitor can reduce elevated total cholesterol levels and / or elevated LDL cholesterol levels and / or elevated triglyceride levels.

[0284] In some embodiments, this application relates to a method for preventing and / or treating and / or controlling metabolic syndrome in a patient, the method comprising administering to the patient an inhibitor of B4GALT1 expression and / or function, wherein the metabolic syndrome is further or independently associated with elevated total cholesterol levels and / or elevated LDL cholesterol levels and / or elevated triglyceride levels, preferably wherein the inhibitor can reduce elevated total cholesterol levels and / or elevated LDL cholesterol levels and / or elevated triglyceride levels.

[0285] In a specific implementation, this application relates to the use of inhibitors of B4GALT1 expression and / or function in the prevention and / or treatment and / or control of metabolic syndrome in patients, wherein the metabolic syndrome is further or independently associated with elevated total blood cholesterol levels, preferably wherein the inhibitor can reduce elevated total blood cholesterol levels.

[0286] In a specific implementation, this application relates to a method for preventing and / or treating and / or controlling metabolic syndrome in a patient, the method comprising administering to the patient an inhibitor of B4GALT1 expression and / or function, wherein the metabolic syndrome is further or independently associated with elevated total blood cholesterol levels, preferably wherein the inhibitor can reduce elevated total blood cholesterol levels.

[0287] In other words, in some implementations, patients who have or are at risk of developing metabolic syndrome have fasting plasma total cholesterol levels of at least 200 mg / dL, at least 205 mg / dL, at least 210 mg / dL, at least 215 mg / dL, at least 220 mg / dL, at least 225 mg / dL, at least 230 mg / dL, at least 235 mg / dL, or at least 240 mg / dL.

[0288] Preferably, administering an inhibitor of B4GALT1 expression and / or function to patients with or at risk of developing metabolic syndrome can reduce the patients' total cholesterol levels, thereby preventing, alleviating, or reversing metabolic syndrome. For example, treating patients with elevated total cholesterol levels and at risk of developing metabolic syndrome using the inhibitor of this application can prevent the occurrence of metabolic syndrome. Similarly, treating patients with elevated total cholesterol levels and who already have metabolic syndrome using the inhibitor of this application can prevent the worsening or further development of metabolic syndrome, and may even reverse metabolic syndrome.

[0289] In a specific implementation, this application relates to the use of inhibitors of B4GALT1 expression and / or function in the prevention and / or treatment and / or control of metabolic syndrome in patients, wherein the metabolic syndrome is further or independently associated with elevated blood LDL cholesterol levels, preferably wherein the inhibitor can reduce elevated blood LDL cholesterol levels.

[0290] In a specific implementation, this application relates to a method for preventing and / or treating and / or controlling metabolic syndrome in a patient, the method comprising administering to the patient an inhibitor of B4GALT1 expression and / or function, wherein the metabolic syndrome is further or independently associated with elevated blood LDL cholesterol levels, preferably wherein the inhibitor can reduce elevated blood LDL cholesterol levels.

[0291] In other words, in some implementations, patients who have or are at risk of developing metabolic syndrome have fasting plasma LDL cholesterol levels of at least 100 mg / dL, at least 105 mg / dL, at least 110 mg / dL, at least 115 mg / dL, at least 120 mg / dL, at least 125 mg / dL, at least 130 mg / dL, at least 135 mg / dL, at least 140 mg / dL, at least 145 mg / dL, at least 150 mg / dL, at least 155 mg / dL, or at least 160 mg / dL.

[0292] Preferably, administering an inhibitor of B4GALT1 expression and / or function to patients with or at risk of developing metabolic syndrome can reduce the patients' serum LDL cholesterol levels, thereby preventing, alleviating, or reversing metabolic syndrome. For example, treating patients with elevated serum LDL cholesterol levels and at risk of developing metabolic syndrome using the inhibitor of this application can prevent the occurrence of metabolic syndrome. Similarly, treating patients with elevated serum LDL cholesterol levels and who already have metabolic syndrome using the inhibitor of this application can prevent the worsening or further development of metabolic syndrome, and may even reverse metabolic syndrome.

[0293] In a specific implementation, this application relates to the use of inhibitors of B4GALT1 expression and / or function in the prevention and / or treatment and / or control of metabolic syndrome in patients, wherein the metabolic syndrome is further or independently associated with elevated blood triglyceride levels, preferably wherein the inhibitor can reduce elevated blood triglyceride levels.

[0294] In a specific implementation, this application relates to a method for preventing and / or treating and / or controlling metabolic syndrome in a patient, the method comprising administering to the patient an inhibitor of B4GALT1 expression and / or function, wherein the metabolic syndrome is further or independently associated with elevated blood triglyceride levels, preferably wherein the inhibitor can reduce elevated blood triglyceride levels.

[0295] In other words, in some implementations, patients who have or are at risk of developing metabolic syndrome have fasting plasma triglyceride levels of at least 150 mg / dL, at least 175 mg / dL, at least 200 mg / dL, at least 225 mg / dL, at least 250 mg / dL, at least 275 mg / dL, at least 300 mg / dL, at least 325 mg / dL, at least 350 mg / dL, at least 375 mg / dL, or at least 400 mg / dL.

[0296] Preferably, administering an inhibitor of B4GALT1 expression and / or function to patients with or at risk of developing metabolic syndrome can reduce their serum triglyceride levels, thereby preventing, alleviating, or reversing metabolic syndrome. For example, treating patients with elevated serum triglyceride levels and at risk of developing metabolic syndrome using the inhibitor of this application can prevent the occurrence of metabolic syndrome. Similarly, treating patients with elevated triglyceride levels and who already have metabolic syndrome using the inhibitor of this application can prevent the worsening or further development of metabolic syndrome, and may even reverse metabolic syndrome.

[0297] As discussed in more detail herein, the inhibitors according to this application can be used for the prevention and / or treatment and / or control of metabolic diseases. As used herein, the term "metabolic disease" refers to a disease or condition that affects human metabolic processes.

[0298] Preferably, metabolic disease refers to a disease associated with any of the factors disclosed herein, such as weight gain, obesity, insulin resistance, elevated blood glucose levels, elevated blood insulin levels, elevated blood HbA1c levels, elevated blood free fatty acid levels, elevated blood fibrinogen levels, elevated blood total cholesterol levels, elevated blood LDL cholesterol levels, and / or elevated blood triglyceride levels.

[0299] Patients receiving treatment may be those already suffering from metabolic diseases or those at risk of developing metabolic diseases. That is, in some embodiments, the inhibitors of this application can be used to treat and / or control existing metabolic diseases. Using the inhibitors of this application to treat and / or control existing metabolic diseases can prevent the metabolic diseases from worsening and / or reverse them. In some cases, using the inhibitors of this application to treat existing metabolic diseases can even cure them. In some embodiments, the inhibitors of this application can be used to prevent the development of clinical manifestations of metabolic diseases in patients at risk of developing them.

[0300] Those skilled in the art can diagnose whether a patient has a metabolic disease or is at risk of developing one. For example, a metabolic disease can be diagnosed based on weight gain and / or one or more blood markers disclosed herein. Those skilled in the art are aware of thresholds for one or more blood markers that indicate the presence of a metabolic disease or a risk of developing one.

[0301] In some implementations, the metabolic disease is diabetes, specifically type 2 diabetes (T2D), as defined elsewhere herein.

[0302] In some implementations, the metabolic disease is fatty liver disease, particularly non-alcoholic fatty liver disease (NAFLD). As used herein, “fatty liver disease” refers to a condition characterized by excessive fat accumulation in the liver, which can lead to serious illnesses such as chronic hepatitis and cirrhosis. In patients with fatty liver disease, lipids, especially triglycerides, accumulate in hepatocytes at levels exceeding physiological limits. Biochemically, the criterion for diagnosing fatty liver is that triglycerides constitute approximately 10% (100 mg / g wet weight) or more of the liver tissue's wet weight. Fatty liver disease is typically detected by observing elevated levels of liver-specific enzymes in serum (such as ALT and AST transaminases, indicators of hepatocyte damage) and the presence of symptoms including fatigue and pain in the liver area, but a definitive diagnosis usually requires a biopsy. As used herein, the terms “NAFLD” or “non-alcoholic fatty liver disease” refer to a condition characterized by fat deposition (steatodegeneration) in the liver not caused by excessive alcohol consumption. This disease is associated with insulin resistance and metabolic syndrome.

[0303] In some embodiments, the metabolic disease is nonalcoholic steatohepatitis (NASH). As used herein, the term “NASH” refers collectively to a state in which the liver develops a pathological condition (e.g., inflammation, ballooning degeneration, fibrosis, cirrhosis, or cancer), or a state in which the liver may be susceptible to such pathological conditions. Furthermore, “NASH” is distinct from “simple steatosis,” which refers to the accumulation of fat solely in the liver without developing into other conditions that predispose to liver disease.

[0304] In some embodiments, the metabolic disease is metabolic syndrome. According to this application, the term "metabolic syndrome" as used herein refers to a range of factors (metabolic abnormalities) associated with an increased risk of cardiovascular disease, such as hypertension, obesity, hyperlipidemia, diabetes, central obesity, hyperglycemia, and hepatic steatosis. Metabolic syndrome is becoming increasingly common, largely due to the rising prevalence of obesity. The International Diabetes Federation defines metabolic syndrome as central obesity (body mass index > 30 kg / m²). 2 And two or more of the following: 1) Triglycerides >150 mg / dL; 2) High-density lipoprotein (HDL) <40 mg / kL for men and <50 mg / dL for women, or currently receiving specific treatment for low HDL; 3) Elevated blood pressure (BP), such as systolic blood pressure >130 mm Hg or diastolic blood pressure >85 mm Hg, or currently receiving treatment for elevated blood pressure, or previously diagnosed with elevated blood pressure; 4) Fasting blood glucose >100 mg / dL or previously diagnosed with type 2 diabetes.

[0305] In some implementations, the metabolic disease is obesity. As used herein, the term "obesity" refers to a state in which the natural energy reserves stored in the adipose tissue of an animal (specifically humans and other mammals) increase to a certain extent, resulting in an increased risk of certain health conditions or mortality. As used herein, "obesity" is defined for adults as a body mass index (BMI) greater than 30. Obesity is generally associated with excessive weight gain, specifically dietary-induced weight gain. "Dietary-induced weight gain" is defined herein as weight gain resulting from excessive dietary intake, including excessive intake of fat, particularly saturated fat, and optionally excessive intake of monosaccharides, including sucrose and fructose. For a given individual, excessive dietary intake (particularly fat intake and optionally monosaccharide intake) refers to an intake of dietary (particularly fat and optionally monosaccharides) exceeding the amount required to meet their physiological needs and maintain their energy balance. The effectiveness of treatment in reducing or preventing diet-induced weight gain can be assessed by comparing the weight gain of treated subjects with that of the same untreated subjects with the same diet and level of physical activity.

[0306] This study has demonstrated that, compared with mice not receiving siRNA molecules, reducing B4GALT1 mRNA expression with siRNA molecules significantly reduced weight gain in mice fed a high-calorie diet (see Example 9 and...). Figure 12 Therefore, surprisingly, inhibiting the expression and / or function of B4GALT1 can prevent weight gain and / or help control weight gain.

[0307] In some embodiments, when a patient responds to treatment with the inhibitor described in this application and experiences a weight loss, it can be determined that the inhibitor of this application can control and / or reduce the patient's weight gain. For example, during appropriate treatment, the patient's weight may decrease by at least 1%, at least 2%, at least 3%, at least 4%, at least 5%, at least 6%, at least 7%, at least 8%, at least 9%, or at least 10%.

[0308] In some implementations, a patient is considered at risk of obesity if their BMI is greater than 25. In some implementations, a patient is considered obese if their BMI is greater than 30.

[0309] In some embodiments, this application relates to inhibitors of B4GALT1 expression and / or function, suitable for or used in the prevention and / or treatment and / or control of weight gain in patients, wherein the patients are characterized by a BMI ≥ 25, preferably ≥ 30.

[0310] In some embodiments, this application relates to inhibitors of B4GALT1 expression and / or function, suitable for or used in the prevention and / or treatment and / or control of obesity in patients, wherein the patients are characterized by a BMI ≥ 25, preferably ≥ 30.

[0311] The term “body mass index” as used in this article refers to weight (in kilograms) divided by the square of height (in meters).

[0312] In some embodiments, the inhibitors according to this application may be used for the prevention and / or treatment and / or control of vascular diseases. As used herein, the term "vascular disease" refers to any disease, condition, or condition affecting the vascular system, including the heart and blood vessels. Vascular diseases include, but are not limited to, cardiovascular diseases, cerebrovascular diseases, peripheral vascular diseases, atherosclerosis, and atherosclerotic vascular diseases.

[0313] Preferably, the vascular disease refers to any vascular disease associated with any of the factors disclosed herein, such as insulin resistance, elevated blood glucose levels, elevated blood insulin levels, elevated blood HbA1c levels, elevated blood free fatty acid levels, elevated blood fibrinogen levels, elevated blood total cholesterol levels, elevated blood LDL cholesterol levels, elevated blood triglyceride levels, and / or diabetes.

[0314] Preferably, the vascular disease refers to any vascular disease associated with any of the factors disclosed herein, such as insulin resistance, elevated blood free fatty acid levels, elevated blood fibrinogen levels, elevated blood total cholesterol levels, elevated blood LDL cholesterol levels, elevated blood triglyceride levels, and / or diabetes.

[0315] The patients to be treated may be those who already have vascular disease or those at risk of developing vascular disease. That is, in some embodiments, the inhibitors of this application can be used to treat and / or control existing vascular disease. Using the inhibitors of this application to treat and / or control existing vascular disease can prevent the disease from worsening and / or reverse it. In some cases, using the inhibitors of this application to treat existing vascular disease can even cure it. In some embodiments, the inhibitors of this application can be used to prevent the development of clinical manifestations of vascular disease in patients at risk of developing it.

[0316] Those skilled in the art can diagnose whether a patient has vascular disease or is at risk of developing vascular disease. For example, vascular disease can be diagnosed based on one or more blood markers disclosed herein. Those skilled in the art are aware of thresholds for one or more blood markers that indicate the presence of vascular disease or a risk of developing vascular disease. Furthermore, various imaging techniques can be used to examine the heart or blood vessels.

[0317] This document preferentially defines vascular disease as cardiovascular disease. The term "cardiovascular disease" as used herein refers to diseases affecting the heart or blood vessels, or both, including but not limited to: hypercholesterolemia, atherosclerosis, coronary artery disease, and cerebral diseases such as myocardial infarction, secondary myocardial infarction, myocardial ischemia, angina pectoris, congestive heart disease, cerebral infarction, cerebral thrombosis, cerebral ischemia, and transient ischemic attack. In some embodiments, the vascular disease is atherosclerosis. The term "atherosclerosis" as used herein encompasses vascular diseases and conditions recognized and understood by licensed physicians in the relevant medical field. Atherosclerotic cardiovascular disease, coronary artery disease (also known as coronary artery disease or ischemic heart disease), cerebrovascular disease, and peripheral vascular disease are all clinical manifestations of atherosclerosis and are therefore all encompassed under the terms "atherosclerosis" and "atherosclerotic disease." The inhibitors of this application may be administered to prevent or reduce the risk of coronary events, cerebrovascular events, or intermittent claudication, or the risk of recurrence (if the possibility of recurrence exists). Coronary artery disease events are defined as including coronary artery disease death, myocardial infarction (i.e., heart attack), and coronary revascularization. Cerebrovascular events are defined as including ischemic or hemorrhagic stroke (also known as cerebrovascular accident) and transient ischemic attack (TIA). Intermittent claudication is a clinical manifestation of peripheral vascular disease. The term "atherosclerotic disease events" as used in this article is intended to encompass coronary artery disease events, cerebrovascular events, and intermittent claudication. It is intended for individuals who have experienced one or more non-fatal atherosclerotic disease events with a history of recurrence. The term "atherosclerosis-related disorder" should be understood as a disorder related to, caused by, or resulting from atherosclerosis.

[0318] In some embodiments, the inhibitors used to treat cardiovascular disease or atherosclerosis are ligand-conjugated double-stranded siRNAs, more preferably conjugated to the target ligands disclosed herein. Therefore, in specific embodiments, this application relates to the use of inhibitors of B4GALT1 expression and / or function in controlling and / or treating and / or preventing cardiovascular disease or atherosclerosis, wherein the inhibitors of B4GALT1 expression and / or function are siRNAs conjugated to target ligands.

[0319] Inhibitors The inhibitors of this application include nucleic acids such as siRNA, antibodies and their antigen-binding fragments, such as monoclonal antibodies, peptides, antibody-drug conjugates, and small molecules. Nucleic acids such as siRNA are preferred.

[0320] The inhibitor of this application may be an inhibitor of B4GALT1 expression and / or function. That is, in some embodiments, the inhibitor may be an inhibitor of B4GALT1 expression in cells. In some embodiments, the inhibitor may inhibit the function of the B4GALT1 enzyme.

[0321] Preferably, the inhibitor of this application inhibits B4GALT1 expression, thereby causing B4GALT1 mRNA knockdown. Knockdown of B4GALT1 mRNA is preferably achieved through hybrid nucleic acid, such as siRNA.

[0322] The B4GALT1 gene is expressed in multiple cell types / tissues in the human body. Therefore, the inhibitor of this application does not necessarily need to target a specific cell type / tissue. However, blood glucose levels and lipid metabolism are primarily controlled by the liver. Therefore, targeting the liver (specifically hepatocytes) with the inhibitor of this application may be beneficial in achieving the therapeutic effects disclosed herein. Therefore, in a specific embodiment, this application relates to a B4GALT1 expression inhibitor, wherein the inhibitor causes hepatocyte-specific B4GALT1 knockdown.

[0323] Those skilled in the art are aware of methods for delivering inhibitors to the liver, particularly hepatocytes. For example, inhibitors can be injected directly into the liver. However, it is preferred herein that the inhibitors be chemically modified with ligands to improve or achieve targeting of the liver. For example, the inhibitors of this application can be chemically modified with ligands of receptors expressed on hepatocytes, such as hepatocyte-specific desialyl glycoprotein receptors (ASGPR). For hepatocytes expressing ASGPR, efficient targeting can be achieved by conjugating the inhibitor to one or more GalNAc residues or derivatives thereof (as defined in more detail elsewhere herein).

[0324] The following describes some preferred features when the inhibitor in this application is an oligonucleotide (such as siRNA).

[0325] In some embodiments, the nucleic acid comprises a first strand containing a sequence at least partially complementary to a portion of the RNA transcribed from the B4GALT1 gene (SEQ ID NO:1). In a preferred embodiment, the nucleic acid comprises a first strand containing a sequence at least partially complementary to B4GALT1 mRNA (NM_001497.4).

[0326] In some embodiments, the nucleic acid used to inhibit B4GALT1 expression includes a double-stranded region comprising a first strand and a second strand at least partially complementary to the first strand, wherein the first strand: (i) is at least partially complementary to a portion of the RNA transcribed from the B4GALT1 gene, and (ii) includes at least 17 consecutive nucleosides, wherein the nucleosides differ from any sequence in SEQ ID NO:22-41 or SEQ ID NO:202-301 by 0 or 1 nucleoside.

[0327] In some embodiments, the first chain contains nucleosides at positions 2-18 of any sequence listed in SEQ ID NO:22-41 or SEQ ID NO:202-301.

[0328] In some embodiments, the nucleic acid used to inhibit B4GALT1 expression includes a double-stranded region comprising a first strand and a second strand at least partially complementary to the first strand, wherein the first strand: (i) is at least partially complementary to a portion of the RNA transcribed from the B4GALT1 gene, and (ii) includes at least 21 consecutive nucleosides, wherein the nucleosides differ from any sequence in SEQ ID NO:202-301 by 0 or 1 nucleoside.

[0329] In some embodiments, the first chain contains nucleosides at positions 2-22 of any of the sequences listed in SEQ ID NO:202-301.

[0330] In some embodiments, the first chain comprises either SEQ ID NO:22-41 or SEQ ID NO:202-301.

[0331] In some embodiments, the second chain comprises a nucleoside sequence of at least 17 consecutive nucleosides, the nucleoside sequence differing from any of the sequences in SEQ ID NO:42-61 or SEQ ID NO:302-401 by 0 or 1 nucleoside; wherein the second chain has at least 85% complementarity with the first chain in the 17 consecutive nucleoside region.

[0332] In some embodiments, the second chain comprises a nucleoside sequence of at least 19 consecutive nucleosides, the nucleoside sequence differing from any of the sequences in SEQ ID NO:302-401 by 0 or 1 nucleoside; wherein the second chain has at least 85% complementarity with the first chain in the 19 consecutive nucleoside region.

[0333] In some embodiments, the second chain comprises a nucleoside sequence of at least 21 consecutive nucleosides, the nucleoside sequence differing from any of the sequences in SEQ ID NO:302-401 by 0 or 1 nucleoside; wherein the second chain has at least 85% complementarity with the first chain within the 21 consecutive nucleoside regions.

[0334] In some embodiments, the second chain comprises any one of the sequences in SEQ ID NO:42-61 or SEQ ID NO:302-401.

[0335] In some embodiments, the nucleic acid comprises a first strand and a second strand; wherein the first strand comprises, consists of, or is substantially composed of nucleoside sequences, the nucleoside sequences differing from any of the sequences in SEQ ID NO:22-41 or SEQ ID NO:202-301 by 0 or 1 nucleoside. The second chain contains, consists of, or is substantially composed of nucleoside sequences, which differ from any of the sequences in SEQ ID NO:42-61 or SEQ ID NO:302-401 by 0 or 1 nucleoside.

[0336] The preferred dual-chain region described herein is formed between a first (antonymous) chain and a complementary second (just) chain. Table 2 below lists exemplary pairs of complementary antisense chains and just chains.

[0337] Table 2

[0338] In a specific implementation, this application relates to a nucleic acid comprising a first strand and a second strand, wherein the first strand and the second strand comprise, consist of, or are substantially composed of nucleoside sequences, wherein the nucleoside sequences differ from any of the following first and second sequences by 0 or 1 nucleoside:

[0339] In some embodiments, the nucleic acid used to inhibit B4GALT1 expression includes a double-stranded region comprising a first strand and a second strand at least partially complementary to the first strand, wherein the first strand: (i) is at least partially complementary to a portion of the RNA transcribed from the B4GALT1 gene, and (ii) includes at least 17 consecutive nucleosides whose sequences differ from any of the sequences in SEQ ID NO:62-81 or SEQ ID NO:402-513 by 0 or 1 nucleoside.

[0340] In some embodiments, the first chain contains nucleosides at positions 2-18 of any sequence listed in SEQ ID NO:62-81 or SEQ ID NO:402-513.

[0341] In some embodiments, the nucleic acid used to inhibit B4GALT1 expression includes a double-stranded region comprising a first strand and a second strand at least partially complementary to the first strand, wherein the first strand: (i) is at least partially complementary to a portion of the RNA transcribed from the B4GALT1 gene, and (ii) includes at least 21 consecutive nucleosides whose sequences differ from any of the sequences in SEQ ID NO:402-513 by 0 or 1 nucleoside.

[0342] In some embodiments, the first chain contains nucleosides at positions 2-22 of any of the sequences listed in SEQ ID NO:402-513.

[0343] In some embodiments, the first chain comprises any one of the sequences in SEQ ID NO:62-81 or SEQ ID NO:402-513.

[0344] The modification patterns of the nucleic acids listed in SEQ ID NO:62-81 and SEQ ID NO:402-513 are summarized in Table 3 below: Table 3

[0345] In some embodiments, the second chain comprises a nucleoside sequence of at least 17 consecutive nucleosides, the nucleoside sequence differing from any of the sequences in SEQ ID NO:82-101 or SEQ ID NO:514-621 by 0 or 1 nucleoside; wherein the second chain has at least 85% complementarity with the first chain within the 17 consecutive nucleoside region.

[0346] In some embodiments, the second chain comprises a nucleoside sequence of at least 19 consecutive nucleosides, the nucleoside sequence differing from any of the sequences in SEQ ID NO:514-621 by 0 or 1 nucleoside; wherein the second chain has at least 85% complementarity with the first chain in the 19 consecutive nucleosides.

[0347] In some embodiments, the second chain comprises a nucleoside sequence of at least 21 consecutive nucleosides, the nucleoside sequence differing from any of the sequences in SEQ ID NO:514-621 by 0 or 1 nucleoside; wherein the second chain has at least 85% complementarity with the first chain within the 21 consecutive nucleoside regions.

[0348] In some embodiments, the second chain comprises either SEQ ID NO:82-101 or SEQ ID NO:514-621.

[0349] The modification patterns of the nucleic acids listed in SEQ ID NO:82-101 and SEQ ID NO:514-621 are summarized in Table 4 below: Table 4

[0350] As used herein, particularly in Tables 3 and 4, the following abbreviations are used for modified nucleosides: Am represents 2'-O-methyladenosine, Cm represents 2'-O-methylcytidine, Gm represents 2'-O-methylguanosine, Um represents 2'-O-methyluridine, Af represents 2'-fluoroadenosine, Cf represents 2'-fluorocytidine, Gf represents 2'-fluoroguanosine, and Uf represents 2'-fluorouridine.

[0351] Furthermore, the letter "s" is an abbreviation for the thiophosphate bond between two consecutive (modified) nucleosides. For example, the abbreviation "AmsAm" is used to represent two consecutive 2'-O-methyladenosine nucleosides linked by a 3'-5' thiophosphate bond. Nucleosides linked by a standard 3'-5' phosphodiester bond do not use this abbreviation. For example, the abbreviation "AmAm" is used to represent two consecutive 2'-O-methyladenosine nucleosides linked by a 3'-5' phosphodiester bond.

[0352] In some embodiments, the nucleic acid comprises a first strand and a second strand; the first strand comprises, consists of, or is substantially composed of a (modified) nucleoside sequence, wherein the (modified) nucleoside sequence differs from any of the sequences in SEQ ID NO:62-81 or SEQ ID NO:402-513 by 0 or 1 nucleoside. The second chain comprises, consists of, or is substantially composed of a (modified) nucleoside sequence, wherein the (modified) nucleoside sequence differs from any of the sequences in SEQ ID NO:82-101 or SEQ ID NO:514-621 by 0 or 1 nucleoside.

[0353] The preferred complementary modification antonym (first) chain and the positive (second) chain are shown in Table 5 below: Table 5

[0354] In a particularly preferred embodiment, this application relates to a nucleic acid comprising a first strand and a second strand, the first strand and the second strand comprising, consisting of, or substantially consisting of nucleoside sequences, said nucleoside sequences differing from any of the following first and second sequences by 0 or 1 nucleoside:

[0355] If there is any ambiguity between the sequences in this specification and the sequences in the attached sequence list, the sequences provided herein shall be considered correct sequences.

[0356] non-base nucleotides In some embodiments, the nucleic acid according to this application contains one, for example two, for example three, for example four or more abasic nucleosides. Abasic nucleosides are modified nucleosides because they lack a base that normally appears at position 1 of the glycosyl moiety. Typically, the abasic nucleosides present in the nucleic acid according to this application have a hydrogen atom at position 1 of the glycosyl moiety.

[0357] The abasic nucleoside is located in the terminal region of the second chain, preferably within the last five nucleosides at the end of the chain. The terminal region may consist of the last five nucleosides containing the abasic nucleoside.

[0358] The second chain may include the following preferred features (unless mutually exclusive, all are particularly considered for combined use): Two or more abase-free nucleotides in the terminal region of the second strand; and / or Two or more abase-free nucleotides in the 5' or 3' terminal region of the second strand; and / or Two or more abasic nucleotides in the 5' or 3' terminal region of the second strand, wherein the abasic nucleotides are present at the overhang described herein; and / or Two or more consecutive abasic nucleosides in the terminal region of the second chain, wherein, preferably, one such abasic nucleoside is a terminal nucleoside; and / or Two or more consecutive abasic nucleotides in the 5' or 3' terminal region of the second strand, wherein, preferably, one such abasic nucleotide is the terminal nucleotide of the 5' or 3' terminal region; and / or A reverse nucleoside bond connecting at least one base-free nucleoside in the terminal region of the second chain to an adjacent base nucleoside; and / or A reverse nucleoside bond connecting at least one base-free nucleotide in the 5' or 3' terminal region of the second chain to an adjacent base nucleotide; and / or The penultimate nucleoside, being a base-free nucleoside, is linked to a non-terminal nucleoside (referred to herein as the penultimate nucleoside) via a reverse bond; and / or When the chain is read along the direction toward the end containing the terminal nucleoside, the baseless nucleoside is connected by a 5'-3' bond as two terminal nucleosides; When the chain is read along the direction toward the end containing the terminal nucleoside, the baseless nucleoside is connected by 3'-5' bonds as two terminal nucleosides; As a nucleoside with no bases at the two ends, wherein the penultimate nucleoside is connected to the penultimate nucleoside via a reverse bond, and wherein the reverse bond is a 5'-5' reverse bond or a 3'-3' reverse bond; As a nucleoside without bases at the two terminal positions, the penultimate nucleoside is linked to the penultimate nucleoside via a reverse bond, and one of the following two cases exists: (1) When read along the end containing the terminal and the penultimate abasic nucleotide, the reverse bond is a 5'-5' reverse bond, and there is a 3'-5' bond between the terminal and the penultimate abasic nucleotide; or (2) When read along the end containing the end and the penultimate abase nucleoside, the reverse bond is a 3'-3' reverse bond and there is a 5'-3' bond between the end and the penultimate abase nucleoside.

[0359] Preferably, the second chain has no nucleoside at its end.

[0360] Preferably, the terminal region of the second chain has two or more abase-free nucleosides, preferably at the terminal position and the penultimate position.

[0361] Preferably, two or more abasic nucleosides are consecutive; for example, all abasic nucleosides can be consecutive. For example, the terminal 1, terminal 2, terminal 3, or terminal 4 nucleotides can be abasic nucleosides.

[0362] Unless there is only one abase-free nucleoside at the end (in which case it is in reverse bond with the adjacent nucleoside), the abase-free nucleoside can also be linked to the adjacent nucleoside through a 5'-3' phosphodiester bond or a reverse bond.

[0363] Reverse bonds (also known as inverted bonds, which are common in the art) include 5'-5', 3'-3', 3'-2', or 2'-3' phosphodiester bonds between adjacent glycosyl moieties of a nucleoside.

[0364] Non-terminal abasic nucleosides have two phosphodiester bonds, each attached to one of the two adjacent nucleosides. These bonds can be anti-bonds or 5'-3 phosphodiester bonds, or one of each.

[0365] A preferred embodiment includes two abase-free nucleosides at the terminal position and the penultimate position of the second chain, wherein the reverse nucleoside bond is located between the penultimate (abase-free) nucleoside and the penultimate nucleoside.

[0366] Preferably, there are two base-free nucleosides at the end and subterminal positions of the second chain, and the subterminal nucleoside is connected to the penultimate nucleoside via an anti-nucleoside internucleotide bond, and is connected to the terminal nucleoside via a 5'-3' or 3'-5' phosphodiester bond (read along the direction of the molecule end).

[0367] The different preferred features are as follows: The reverse nucleoside interbond is a 3'-3' reverse bond. The reverse nucleoside interbond is located in the terminal region distal to the 5' end phosphate group of the second chain.

[0368] The reverse nucleoside interbond is a 5'-5' reverse bond. The reverse nucleoside interbond is located in the terminal region distal to the 3' end hydroxyl group of the second chain.

[0369] In some embodiments, the second chain comprises two consecutive abasic nucleosides in its 5' terminal region, one of which is a terminal nucleoside in the 5' terminal region of the second chain, and the other is a penultimate nucleoside in the 5' terminal region of the second chain, wherein: (a) the penultimate abasic nucleoside is connected to the first adjacent base nucleoside in the adjacent 5' proximal terminal region by a reverse nucleoside bond; and (b) the reverse bond is a 5'-5' reverse bond; and (c) when read along the end containing the terminal and penultimate abasic nucleosides, there is a 3'-5' bond between the terminal and penultimate abasic nucleosides. More typically, (i) both the first and second chains are 19 or 23 nucleotides in length; (ii) each of the three consecutive positions in the 5' proximal region of the second chain has two thiophosphate nucleotide inter-bonds, wherein the first thiophosphate nucleotide inter-bond exists between the adjacent first base nucleotide of (a) and the adjacent second base nucleotide in the 5' proximal region of the second chain, and the second thiophosphate nucleotide inter-bond exists between the adjacent second base nucleotide and the adjacent third base nucleotide in the 5' proximal region of the second chain; (ii) (i) Two thiophosphate nucleoside bonds are present between three consecutive positions in the 5' and 3' end regions of the first strand, such that each terminal nucleoside in the 5' and 3' end regions of the first strand is connected to the corresponding penultimate 5' and 3' nucleoside via a thiophosphate nucleoside bond, and each penultimate 5' and 3' nucleoside is connected to the corresponding penultimate 5' and 3' nucleoside via a thiophosphate nucleoside bond; and (iv) the second strand of the nucleic acid is conjugated directly or indirectly to one or more ligands in the 3' end region of the second strand.

[0370] Alternatively, the second chain comprises two consecutive abasic nucleosides, preferably located at the protruding end of the 3' terminal region of the second chain, one of which is the terminal nucleoside of the 3' terminal region of the second chain, and the other is the penultimate nucleoside of the 3' terminal region of the second chain, wherein: (a) the penultimate abasic nucleoside is connected to the first adjacent nucleoside in the adjacent 3' proximal terminal region by a reverse nucleoside bond; and (b) the reverse bond is a 3'-3' reverse bond; and (c) when read along the direction toward the end containing the terminal and penultimate abasic nucleosides, there is a 5'-3' bond between the terminal and penultimate abasic nucleosides. More typically, (i) both the first and second chains are 19 or 23 nucleotides in length; (ii) each of the three consecutive positions in the 3' proximal region of the second chain has two thiophosphate nucleotide bonds, wherein the first thiophosphate nucleotide bond is between the adjacent first nucleotide of (a) and the adjacent second nucleotide in the 3' proximal region of the second chain, and the second thiophosphate nucleotide bond is between the adjacent second nucleotide and the adjacent third nucleotide in the 3' proximal region of the second chain; (iii) Two thiophosphate nucleoside internucleotide bonds are respectively located between three consecutive positions in the 5' and 3' end regions of the first strand, thereby connecting each terminal nucleoside in the 5' and 3' end regions of the first strand to the corresponding penultimate 5' and 3' adjacent nucleoside via thiophosphate nucleoside internucleotide bonds, and connecting each first 5' and 3' penultimate 5' and 3' adjacent third nucleoside via thiophosphate nucleoside internucleotide bonds; and (iv) the second strand of the nucleic acid is conjugated directly or indirectly to one or more ligand moieties in the 5' end region of the second strand.

[0371] The following are structural examples (the specific RNA nucleoside shown is not limited and can be any RNA nucleoside): A. 3'-3' reverse bond (and also shows that the 5'-3 direction of the last phosphodiester bond between the two baseless molecules is towards the end of the molecule) B. Show the 5'-5' reverse bond (and show the 3'-5' orientation of the last phosphodiester bond between two baseless molecules when read towards the end of the molecule). The presence of one or more abasic nucleosides in nucleic acids is provided by the presence of one or more reverse nucleoside bonds (i.e., 5'-5' or 3'-3' reverse nucleoside bonds). The reverse bond arises from a change in the orientation of adjacent nucleoside sugars, giving the sugar a 3'-5' orientation instead of the conventional 5'-3' orientation (refer to the numbering of ring atoms on the nucleoside sugar). The one or more abasic nucleosides present in the nucleic acids of this application preferably include such reverse nucleoside sugars.

[0372] If the terminal nucleoside has a reverse orientation, this will result in the entire nucleic acid exhibiting a "reverse" terminal configuration. Although some structures drawn and cited in this article are represented using the conventional 5'-3' orientation (refer to the numbering of ring atoms on the nucleoside sugar), it is understood that if a terminal nucleoside with a reverse orientation and a proximal 3'-3' reverse bond is present, a nucleic acid with an overall 5'-5' terminal structure will be formed (i.e., a conventional 3' terminal nucleoside becomes a 5' terminal nucleoside). Alternatively, if a terminal nucleoside with a reverse orientation and a proximal 5'-5' reverse bond is present, a nucleic acid with an overall 3'-3' terminal structure will be formed.

[0373] As described herein, a proximal 3'-3' or 5'-5' reverse bond may comprise a reverse bond directly adjacent to / connected to a reverse-oriented terminal nucleotide (e.g., a single reverse-oriented terminal nucleotide). Alternatively, as described herein, a proximal 3'-3' or 5'-5' reverse bond may comprise a reverse bond between two or more adjacent reverse-oriented nucleotides, such as between two or more reverse-oriented terminal nucleotides, such as between the terminal and penultimate nucleotide. In this way, the reverse bond can be connected to the penultimate nucleotide with the reverse orientation. Although those skilled in the art will understand that the reverse orientation described above can result in a nucleic acid molecule as a whole having a 3'-3' or 5'-5' terminal structure as described herein, it should also be understood that if one or more additional reverse bonds and / or reverse-oriented nucleotides are present, the nucleic acid molecule as a whole may have a 3'-5' terminal structure relative to the conventionally positioned 5' / 3' end.

[0374] On the one hand, nucleic acids can have 3'-3' reverse bonds, and the terminal glycosyl moiety can contain a 5' OH instead of a 5' phosphate group at the 5' position of the terminal glycosyl moiety.

[0375] Therefore, those skilled in the art will clearly understand that, compared to the more conventional 5'-3' structure shown herein (refer to the ring atom numbering on the terminal nucleotide sugar), the 5'-5', 3'-3', and 3'-5' (read along the terminal direction) terminal variants are all included within the scope of this disclosure when one or more anti-bonds are present.

[0376] For example, in cases where a reverse nucleotide bond and / or one or more nucleotides with reverse orientations constitute a reverse end, when the relative position of the bond (e.g., with a linker) or the position of an internal feature (e.g., a modified nucleotide) is defined relative to the 5' or 3' end of the nucleic acid, the 5' or 3' end refers to the conventional 5' or 3' end that would be present assuming the absence of a reverse bond; where the conventional 5' or 3' end is determined by considering the directionality of most nucleotide bonds within the nucleic acid and / or the orientation of the nucleotides. Based on these internal linking bonds and / or the orientation of the nucleotides, it can be determined which ends of the nucleic acid would constitute the conventional 5' or 3' end of the molecule in the absence of a reverse bond (refer to the ring atom numbering on the terminal nucleotide sugar).

[0377] For example, in the structure shown below, the first two positions at the "5'" end have no base residues. When the terminal nucleotide has a reverse orientation, the "5'" end (i.e., the conventional 5' end) shown in the diagram below can actually contain a 3'OH, given the reverse nucleotide at the terminal position. Nevertheless, when read according to the standard 5' [PO4] to 3' [OH] orientation of nucleic acid molecules (refer to the ring atom numbering on the nucleotide sugar), the majority of the molecule will still contain conventional internucleotide bonds, i.e., extending from the 3'OH of the sugar to the 5' phosphate group of the next sugar. This connection pattern can be used to identify the conventional 5' and 3' ends that are typically present when there is no reverse end configuration.

[0378] A. 5' AA-Me-Me-Me-Me-Me-Me-F-Me-FFF-Me-Me-Me-Me-Me-Me-Me-Me-Me-Me3' Preferably, the reverse bond is located at the end of the nucleic acid (e.g., RNA), which is located at the distal end of the molecular ligand portion (e.g., the GalNAc portion).

[0379] The GalNAc-siRNA construct with 5'-GalNAc on the positive strand can have a reverse bond at the other end of the positive strand.

[0380] GalNAc-siRNA constructs with 3'-GalNAc on the positive strand can have a reverse bond at the other end of the positive strand.

[0381] Nucleic acid length In one aspect, i) the length of the first strand of nucleic acid is 17-30 nucleosides, preferably 19-25 nucleosides, more preferably 19 or 23 nucleosides; and / or ii) the length of the second strand of nucleic acid is 17-30 nucleosides, preferably 19-25 nucleosides, more preferably 19 or 21 nucleosides.

[0382] Typically, the length of the double-stranded region of a nucleic acid is 17-30 nucleotides, more preferably 19 or 21 nucleotides. Similarly, the length of the complementary region between the first strand and the portion of RNA transcribed from the B4GALT1 gene is 17-30 nucleotides.

[0383] In one aspect, i) the length of the first strand of the nucleic acid is 15-30 nucleotides, preferably 19-25 nucleotides, more preferably 23 or 25 nucleotides; and / or ii) The length of the second strand of the nucleic acid is 15-30 nucleosides, preferably 19-25 nucleosides, and more preferably 23 nucleosides.

[0384] Generally, the double-stranded structure of nucleic acids (such as iRNA) is approximately 15-30 base pairs in length, for example, lengths of 15-29, 15-28, 15-27, 15-26, 15-25, 15-24, 15-23, 15-22, 15-21, 15-20, 15-19, 15-18, 15-17, 18-30, 18-29, 18-28, 18-27, 18-26, 18-25, 18-24, 18-23, 18-22, 18-21, 18-20, 1... 9-30, 19-29, 19-28, 19-27, 19-26, 19-25, 19-24, 19-23, 19-22, 19-21, 19-20, 20-30, 20-29, 20-28, 20-27, 20-26, 20-25, 20-24, 20-23, 20-22, 20-21, 21-30, 21-29, 21-28, 21-27, 21-26, 21-25, 21-24, 21-23, or 21-22 base pairs. The range and length between the above ranges and lengths are also considered part of this application.

[0385] Similarly, the length of the complementary regions between the antisense sequence and the target sequence and / or between the antisense sequence and the sense sequence is approximately 15-30 nucleotides, for example, lengths of 15-29, 15-28, 15-27, 15-26, 15-25, 15-24, 15-23, 15-22, 15-21, 15-20, 15-19, 15-18, 15-17, 18-30, 18-29, 18-28, 18-27, 18-26, 18-25, 18-24, 18-23, 18-22, 18-21. 18-20, 19-30, 19-29, 19-28, 19-27, 19-26, 19-25, 19-24, 19-23, 19-22, 19-21, 19-20, 20-30, 20-29, 20-28, 20-27, 20-26, 20-25, 20-24, 20-23, 20-22, 20-21, 21-30, 21-29, 21-28, 21-27, 21-26, 21-25, 21-24, 21-23, or 21-22 nucleosides. The range and length between the above ranges and lengths are also considered part of this application.

[0386] In some preferred embodiments, the length of the complementary region between the antisense sequence and the target sequence and / or the complementary region between the antisense sequence and the sense sequence is at least 17 nucleotides. For example, the length of the complementary region between the antisense strand and the target site is 19-21 nucleotides, for example, the length of the complementary region is 21 nucleotides.

[0387] In a preferred embodiment, the length of each chain does not exceed 30 nucleosides.

[0388] In some preferred embodiments, the double-stranded structure of the nucleic acid (such as siRNA) is 19 base pairs in length. In a particularly preferred embodiment, the double-stranded structure may have one of the following structures: or .

[0389] The nucleic acids (such as dsRNA) described herein may also contain one or more single-stranded nucleoside overhangs, such as 1-4, 2-4, 1-3, 2-3, 1, 2, 3, or 4 nucleosides. The nucleoside overhangs may contain or be composed of nucleosides / nucleoside analogs (including deoxynucleosides / nucleosides). The overhangs may be located on the sense strand, the antisense strand, or any combination thereof. Furthermore, the nucleosides of the overhangs may be at the 5' end, 3' end, or both ends of the antisense or sense strand of the nucleic acid (such as dsRNA).

[0390] In some preferred embodiments, at least one chain contains a 3' overhang of at least one nucleoside (e.g., at least one chain contains a 3' overhang of at least two nucleosides). The overhang is suitably located on the antisense / leader chain and / or the sense / follower chain.

[0391] Nucleic acid modification In some embodiments, the nucleic acid (e.g., RNA), such as dsiRNA, of this application does not contain further modifications, such as chemical modifications or conjugations known in the art and described herein.

[0392] In other preferred embodiments, the nucleic acid (e.g., RNA), such as dsiRNA, of this application is further chemically modified to increase stability or other beneficial properties.

[0393] In some embodiments of this application, virtually all nucleosides are modified.

[0394] The nucleic acids involved in this application can be synthesized or modified by methods well known in the art, such as those in "Currentprotocols in nucleic acid chemistry," Beaucage, SL et al. (Edrs.), JohnWiley & Sons, Inc., New York, NY, USA, the contents of which are incorporated herein by reference.

[0395] Modifications include, for example, terminal modifications such as 5'-end modifications (phosphorylation, conjugation, reverse bond) or 3'-end modifications (conjugation, DNA nucleoside within RNA or RNA nucleoside within DNA, reverse bond, etc.); base modifications such as substitution with stable bases, unstable bases or bases that can bind to more pairing partners, conjugated bases; sugar modifications (e.g. at the 2'-position or 4'-position) or sugar substitutions; or backbone modifications, including modifications or substitutions of phosphodiester bonds.

[0396] Specific examples of nucleic acid compounds, such as siRNA, used in the embodiments described herein include, but are not limited to, RNA containing a modified backbone or non-natural nucleoside internucleotide bonds. Nucleic acids (e.g., RNA) with a modified backbone comprise nucleic acids that do not have phosphorus atoms in their backbone. For the purposes of this specification, and as sometimes cited in the art, modified nucleic acids, such as RNA, that do not have phosphorus atoms in their nucleoside internucleotide backbone may also be considered oligonucleotides. In some embodiments, modified nucleic acids, such as siRNA, have phosphorus atoms in their nucleoside internucleotide backbone.

[0397] Modified nucleic acid (e.g., RNA) backbones include, for example, thiophosphates, chiral thiophosphates, dithiophosphates, phosphate triesters, aminoalkyl phosphate triesters, methyl and other alkylphosphonates (including 3'-alkylphosphonates and chiral phosphonates), phosphonites, aminophosphates (including 3'-aminoaminophosphates and aminoalkylaminophosphates), thioaminophosphates, thioalkylphosphonates, thioalkyl phosphate triesters, and borophosphates with normal 3'-5' bonds, 2'-5' linked analogs of these compounds, and those with reverse polarity (where adjacent nucleoside unit pairs are linked at 5'-3' or 5'-2'). Various salts, mixed salts, and free acid forms are also included.

[0398] Modified nucleic acids (such as RNA) may also contain one or more substituted glycosyl groups. Nucleic acids described herein, such as siRNA (e.g., dsiRNA), may contain one of the following at the 2'-position: OH; F; O-, S-, or N-alkyl; O-, S-, or N-alkenyl; O-, S-, or N-alkynyl; or O-alkyl-O-alkyl, wherein the alkyl, alkenyl, and alkynyl groups may be substituted or unsubstituted. Preferred modifications are 2'-O-methyl and 2'-F.

[0399] In some preferred embodiments, the nucleic acid comprises at least one modified nucleoside.

[0400] The nucleic acid of this application may contain one or more modified nucleosides on the first and / or second strand.

[0401] In some implementations, almost all nucleosides of the sense strand and all nucleosides of the antisense strand contain modifications.

[0402] In some implementations, all nucleosides in the sense strand and almost all nucleosides in the antisense strand contain modifications.

[0403] In some implementations, all nucleosides in the sense strand and all nucleosides in the antisense strand contain modifications.

[0404] In one embodiment, at least one of the modified nucleosides is selected from the group consisting of: deoxy-nucleosides, 3'-terminal deoxy-thymidine (dT) nucleosides, 2'-O-methyl modified nucleosides (also referred to herein as 2'-Me, where Me is methoxy), 2'-fluorine modified nucleosides, 2'-deoxy-modified nucleosides, locked nucleosides, open-ring nucleosides, conformation-restricted nucleosides, ethyl-restricted nucleosides, base-free nucleosides, 2'-amino-modified nucleosides, 2'-O-allyl-modified nucleosides, and 2'-methyl ... '-C-alkyl-modified nucleosides, 2'-hydroxy-modified nucleosides, 2'-methoxyethyl-modified nucleosides, 2'-O-alkyl-modified nucleosides, morpholino-modified nucleosides, aminophosphates, nucleosides containing non-natural bases, tetrahydropyran-modified nucleosides, 1,5-dehydrohexyl-modified nucleosides, cyclohexenyl-modified nucleosides, nucleosides containing thiophosphate groups, nucleosides containing methylphosphonate groups, nucleosides containing 5'-phosphates, and nucleosides containing 5'-phosphate mimics. In another embodiment, the modified nucleoside comprises a short sequence of 3'-terminal deoxy-thymidine (dT).

[0405] The nucleoside modification may preferably be from, but is not limited to, LNA, HNA, CeNA, 2-methoxyethyl, 2'-O-alkyl, 2-O-allyl, 2'-C-allyl, 2'-fluoro, 2'-deoxy, 2'-hydroxy, and combinations thereof. In another embodiment, the nucleoside modification is 2'-O-methyl (“2-Me”) or 2'-fluoro.

[0406] A preferred modification is the modification of the 2'-OH group of the ribose, which can be selected from 2'-Me or 2'-F modification.

[0407] The preferred nucleic acid contains one or more nucleosides on the first and / or second strand, which are modified to form modified nucleosides, as follows: A nucleic acid whose modification is located at the 2'-OH group of the ribose, which may be selected from 2'-Me or 2'-F modification.

[0408] A nucleic acid wherein, starting from position 1 of the first strand, the first strand contains a 2'-F modification at position 2, position 6, position 14, or any combination thereof.

[0409] A nucleic acid wherein, starting from position 1 of the second strand, the second strand contains a 2'-F modification at position 7, position 9, position 11, or any combination thereof.

[0410] A nucleic acid wherein, starting from position 1 of the second strand, the second strand contains a 2'-F modification at positions 7, and / or 9, and / or 11, and / or 13.

[0411] A nucleic acid wherein, starting from position 1 of the second strand, the second strand contains a 2'-F modification at positions 7, 9, and 11.

[0412] A nucleic acid wherein both the first and second strands contain 2'-Me and 2'-F modifications.

[0413] A nucleic acid comprising at least one thermally unstable modification suitably located at one or more positions 1 to 9 of the first strand, counting from position 1 on the first strand, and / or at one or more positions on the second strand corresponding to positions 1 to 9 of the first strand, wherein the unstable modification is selected from modified unlocked nucleic acids (UNA) and glycol nucleic acids (GNA), preferably glycol nucleic acids.

[0414] A nucleic acid wherein, starting from position 1 of the second strand, the second strand contains three or more 2'-F modifications at positions 7 to 13, for example, four, five, six or seven 2'-F modifications at positions 7 to 13 of the second strand.

[0415] A nucleic acid wherein, starting from position 1 of the second strand, the second strand contains at least three 2'-Me modifications, such as four, five, or six 2'-Me modifications, at positions 1 to 6.

[0416] A nucleic acid wherein the first strand contains at least five consecutive 2'-Me modifications in the 3' end region, preferably including terminal nucleosides in the 3' end region, or at least one or two terminal nucleosides located within the 3' end region.

[0417] A nucleic acid wherein the first strand contains seven consecutive 2'-Me modifications in the 3' terminal region, preferably terminal nucleotides in the 3' terminal region. A nucleic acid, wherein, starting from position 1 on the first strand, position 7 on the first strand contains at least one thermally unstable modification.

[0418] A nucleic acid, which is an siRNA oligonucleotide, wherein, starting from position 1 of the second strand, the siRNA oligonucleotide contains at least three 2'-F modifications at positions 6 to 12 of the second strand.

[0419] A nucleic acid, which is an siRNA oligonucleotide, wherein, starting from position 1 of the second strand, the second strand contains at least three 2'-Me modifications at positions 1 to 6.

[0420] A nucleic acid, specifically an siRNA oligonucleotide, wherein both the first and second strands contain alternating modification patterns, preferably with completely alternating modification patterns throughout their entire length. Specifically, the first strand is modified with: (i) 2'Me modification on odd-numbered nucleotides starting from position 1 on the first strand, and (ii) 2'F modification on even-numbered nucleotides starting from position 1 on the first strand; the second strand is modified with: (i) 2'F modification on odd-numbered nucleotides starting from position 1 on the second strand, and (ii) 2'Me modification on even-numbered nucleotides starting from position 1 on the second strand. Typically, this completely alternating modification pattern is present in blunt-terminated oligonucleotides, wherein both the first and second strands are 19 or 23 nucleotides in length.

[0421] Position 1 of the first or second strand refers to the nucleoside closest to the end of the nucleic acid (ignoring any baseless nucleosides) and connected to the adjacent nucleoside (at position 2) via a 3'-5' internal bond; the bond referred to here is the linking bond between the backbone sugar moieties and is read in the direction away from the end of the molecule.

[0422] Therefore, "position 1 of the positive strand" is the 5' nearest nucleotide (excluding nucleosides) at the conventional 5' end of the positive strand. Typically, the nucleotide at position 1 of the positive strand will correspond to the 5' nucleotide of the selected target nucleic acid sequence; and more generally, starting from position 1 of the positive strand, its nucleotide sequence corresponds to the nucleotide of the target nucleic acid sequence, while also allowing for acceptable mismatches between sequences.

[0423] The “position 1 of the antisense strand” used in this article refers to the outermost nucleoside at the conventional 5' end of the antisense strand (excluding nucleosides without bases). As mentioned earlier, there are complementary regions between the sense and antisense strands, so the antisense strand will also have complementary regions with the target nucleic acid sequence mentioned above.

[0424] In some embodiments, the nucleic acid (e.g., an RNAi reagent) further comprises at least one thiophosphate or methylphosphonate nucleoside internucleotide bond. For example, the thiophosphate or methylphosphonate nucleoside internucleotide bond may be located at the 3' end or terminal region of one strand (i.e., the sense strand or antisense strand); or at the ends of both strands (i.e., the sense strand and the antisense strand).

[0425] In some embodiments, the thiophosphate or methylphosphonate nucleoside internucleotide bond is located at the 5' end or terminal region of one chain (i.e., the sense chain or the antisense chain); or at the end of two chains (i.e., the sense chain and the antisense chain).

[0426] In some embodiments, the thiophosphate or methylphosphonate nucleoside internucleotide bond is located at the 5' and 3' ends or terminal regions of one chain (i.e., the sense chain or the antisense chain); or at the ends of two chains (i.e., the sense chain and the antisense chain).

[0427] Any nucleic acid may contain one or more phosphate thioester (PS) modifications, such as at least two PS nucleotide bonds at the end of the chain.

[0428] At least one of the oligonucleotide chains preferably contains at least two consecutive thiophosphate modifications in its last three nucleotides.

[0429] Therefore, this application also relates to: the nucleic acid disclosed herein, which contains thiophosphate nucleoside inter-bonds, the thiophosphate nucleoside inter-bonds being located between at least two or three consecutive positions, for example, located in the 5' and / or 3' terminal regions and / or near-terminal regions of the second chain, wherein the near-terminal region is preferably adjacent to the terminal region of the second chain containing one or more base-free nucleosides.

[0430] The nucleic acids disclosed herein contain at least two or three consecutive positions between 5' and / or 3' end regions of their first strand, respectively, thiophosphate nucleoside bonds; preferably, the end positions of the 5' and / or 3' end regions of the first strand are connected to their adjacent positions by thiophosphate nucleoside bonds.

[0431] The nucleic acid chain can be RNA, which contains thiophosphate nucleoside bonds between three adjacent nucleosides located at two ends of abase-free nucleosides.

[0432] Preferably, the nucleic acid is a double-stranded RNA, containing two adjacent abase-free nucleotides at the 5' end of the second strand, and a ligand portion containing one or more GalNAc ligands at the opposite 3' end of the second strand. More preferably, the nucleic acid may also contain phosphate-thioester bonds between nucleotides at positions 3-4 and 4-5 of the second strand, starting from position 1 of the second strand. Even more preferably, the nucleic acid may also contain 2'F modifications at positions 7, 9, and 11 of the second strand.

[0433] Nucleic acids with the following structure ; The preferred modification patterns are as follows: A nucleic acid wherein the first-strand modified nucleoside has the following modification pattern (5'-3'): Me–F–Me–F–Me–F–Me–F–Me–F–Me–F–Me–F–Me–F–Me–F–Me.

[0434] A nucleic acid wherein the modified nucleoside of the second strand has the following modification pattern (5'-3'): F(s)Me(s)F–Me–F–Me–F–Me–F–Me–F–Me–F–Me–F–Me–F–Me–F, or F-Me-F–Me–F–Me–F–Me–F–Me–F–Me–F–Me–F–Me–F(s)Me(s)F; Wherein, (s) is the internucleotide bond between thiophosphate esters.

[0435] A nucleic acid wherein the modified nucleoside of the second strand has the following modification pattern (5'-3'): F–Me–F–Me–F–Me–F–Me–F–Me–F–Me–F–Me–F–Me–F–Me–F.

[0436] A nucleic acid wherein the modified nucleoside of the second strand has the following modification pattern (5'-3'): F(s)Me(s)F–Me–F–Me–F–Me–F–Me–F–Me–F–Me–F–Me–F–Me–F, or F-Me-F–Me–F–Me–F–Me–F–Me–F–Me–F–Me–F–Me–F(s)Me(s)F; Wherein, (s) is the internucleotide bond between thiophosphate esters.

[0437] A nucleic acid wherein the modified nucleoside of the second strand has the following modification pattern (5'-3'): ia–ia-F–Me–F–Me–F–Me–F–Me–F–Me–F–Me–F–Me–F–Me–F–Me–F, or F–Me–F–Me–F–Me–F–Me–F–Me–F–Me–F–Me–F–Me–F–Me–F–ia–ia; Wherein, ia represents a reverse abasic nucleoside. In some embodiments, the reverse abasic nucleoside represented by ia-ia is present at the overhang of the 2-nucleoside.

[0438] A nucleic acid wherein the modified nucleoside of the second strand has any of the following modification patterns (5'-3'): ia–ia–F(s)Me(s)F–Me–F–Me–F–Me–F–Me–F–Me–F–Me–F–Me–F–Me–F, or F–Me–F–Me–F–Me–F–Me–F–Me–F–Me–F–Me–F–Me–F–Me(s)F(s)ia–ia; Wherein, (s) represents a thiophosphate nucleoside internucleotide bond, and ia represents a reverse abasic nucleoside. In some embodiments, the reverse abasic nucleoside represented by ia-ia is located at the overhang of the 2-nucleotide pair.

[0439] structural nucleic acid ; Selected ceremony model below: One type of nucleic acid, among which the second type of modification nucleic acid listed below is included in the following one type of modification model (5'-3'): Me-Me-Me-Me-Me-Me-FFFFF-Me-Me-Me-Me-Me-Me-Me-F-Me–Me, or Me-Me-Me-Me-Me-FF-Me-FFFF-Me-Me-Me-Me-Me-Me-Me-Me–Me, or Me-Me-Me-Me-Me-Me-F-Me-FFFF-Me-Me-Me-Me-Me-Me-Me-Me–Me, or Me-Me-Me-Me-Me-Me-Me-Me-FFF-Me-Me-Me-Me-Me-Me-Me-Me-Me–Me, someone Me-Me-Me-Me-Me-Me-F-Me-FFF-Me-Me-Me-Me-Me-Me-Me-Me-Me–Me.

[0440] One type of nucleic acid, among which the second type of modification nucleic acid listed below is included in the following one type of modification model (5'-3'): Me(s)Me(s)Me-Me-Me-Me-FFFFF-Me-Me-Me-Me-Me-Me-Me-F-Me–Me, someone Me(s)Me(s)Me-Me-Me-FF-Me-FFFF-Me-Me-Me-Me-Me-Me-Me-Me–Me, someone Me(s)Me(s)Me-Me-Me-Me-F-Me-FFFF-Me-Me-Me-Me-Me-Me-Me-Me–Me, someone Me(s)Me(s)Me-Me-Me-Me-Me-Me-FFF-Me-Me-Me-Me-Me-Me-Me-Me-Me–Me, or Me(s)Me(s)Me - Me - Me - Me - F - Me - F - F - F - Me - Me - Me - Me- Me - Me - Me - Me - Me – Me, someone Me – Me - Me - Me - Me - Me - F - F - F - F - F - Me - Me - Me - Me -Me - Me - Me - F(s)Me(s)Me, or Me – Me - Me - Me - Me - F - F - Me - F - F - F - F - Me - Me - Me -Me - Me - Me - Me(s)Me(s)Me, or Me – Me - Me - Me - Me - Me -F- Me - F - F - F - F - Me - Me - Me -Me - Me - Me - Me(s)Me(s)Me, or Me – Me - Me - Me - Me - Me - Me - Me - F - F - F - Me - Me - Me - Me- Me - Me - Me - Me(s)Me(s)Me, or Me – Me - Me - Me - Me - Me - F - Me - F - F - F - Me - Me - Me - Me- Me - Me - Me - Me(s)Me(s)Me, where (s) is a phosphorothioate internucleoside bond.

[0441] A nucleic acid, wherein the modified nucleosides of the second strand include any of the following modification patterns (5’-3’): ia–ia-Me-Me-Me-Me-Me-Me-F-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-F-Me–Me, or ia – ia - Me - Me - Me - Me - Me - F - F - Me - F - F - F - F - Me -Me - Me - Me - Me - Me - Me - Me – Me, or ia – ia - Me - Me - Me - Me - Me - Me -F- Me - F - F - F - F - Me -Me - Me - Me - Me - Me - Me - Me – Me, or ia – ia - Me - Me - Me - Me - Me - Me - Me - Me - F - F - F - Me - Me- Me - Me - Me - Me - Me - Me - Me – Me, or ia – ia - Me - Me - Me - Me - Me - Me - F - Me - F - F - F - Me - Me- Me - Me - Me - Me - Me - Me - Me – Me, or Me - Me - Me - Me - Me - Me - F - F - F - F - F - Me - Me - Me - Me -Me - Me - Me - F - Me – Me - ia – ia, or Me - Me - Me - Me - Me - F - F - Me - F - F - F - F - Me - Me - Me -Me - Me - Me - Me - Me – Me - ia – ia, or Me - Me - Me - Me - Me - Me -F- Me - F - F - F - F - Me - Me - Me -Me - Me - Me - Me - Me – Me - ia – ia, or Me - Me - Me - Me - Me - Me - Me - Me - F - F - F - Me - Me - Me - Me- Me - Me - Me - Me - Me – Me - ia – ia, or Me - Me - Me - Me - Me - Me - F - Me - F - F - F - Me - Me - Me - Me- Me - Me - Me - Me - Me – Me- ia – ia, wherein, ia represents an inverted abasic nucleoside, and when the inverted abasic nucleoside represented by ia-ia is present at the 3'-end of the second strand, the inverted abasic nucleoside exists in the form of a 2-nucleotide overhang.

[0442] A nucleic acid, wherein the modified nucleosides of the second strand include any one of the following modification patterns (5'-3'): ia – ia - Me(s)Me(s)Me - Me - Me - Me - Me - Me - F - F - F - F - Me - Me - Me - Me - Me - Me - Me - F - Me - Me, or ia – ia - Me(s)Me(s)Me - Me - Me - F - F - Me - F - F - F - F - Me - Me - Me - Me - Me - Me - Me - Me - Me, or ia – ia - Me(s)Me(s)Me - Me - Me - Me - F - Me - F - F - F - F - Me - Me - Me - Me - Me - Me - Me - Me - Me, or ia – ia - Me(s)Me(s)Me - Me - Me - Me - Me - Me - F - F - F - Me - Me - Me - Me - Me - Me - Me - Me - Me – Me, or ia – ia - Me(s)Me(s)Me - Me - Me - Me - F - Me - F - F - F - Me - Me - Me - Me - Me - Me - Me - Me - Me, or Me – Me - Me - Me - Me - Me - F - F - F - F - F - Me - Me - Me - Me - Me - Me - Me - F(s)Me(s)Me - ia – ia, or Me – Me - Me - Me - Me - F - F - Me - F - F - F - F - Me - Me - Me - Me - Me - Me - Me - Me(s)Me(s)Me - ia – ia, or Me – Me - Me - Me - Me - Me - F - Me - F - F - F - F - Me - Me - Me - Me - Me - Me - Me - Me(s)Me(s)Me - ia – ia, or Me – Me - Me - Me - Me - Me - Me - Me - F - F - F - Me - Me - Me - Me- Me - Me - Me - Me(s)Me(s)Me - ia – ia, or Me - Me - Me - Me - Me - Me - F - Me - F - F - F - Me - Me - Me - Me- Me - Me - Me - Me(s)Me(s)Me - ia – ia, Wherein, (s) is a thiophosphate nucleoside bond, ia represents a reverse abasic nucleoside, and when the reverse abasic nucleoside represented by ia-ia is present at the 3' end of the second chain, the reverse abasic nucleoside exists as two nucleoside overhangs.

[0443] A nucleic acid wherein the modified nucleoside includes any of the following modification patterns: Modification pattern 1: Second strand (5'-3'): Me - Me - Me - Me - Me - Me - F - F - F - F- F - Me - Me - Me - Me - Me - Me - Me - F - Me - Me, First strand (5'-3'): Me - F -Me - F - Me - F - Me - F - F - Me - Me - Me - Me - F - Me - F - Me - Me - Me- Me - Me - Me - Me; Or modification mode 2: second strand (5'-3'): Me - Me - Me - Me - Me - F - F - Me - F- F - F - F - Me - Me - Me - Me - Me - Me - Me - Me - Me, first strand (5'-3'): Me -F - Me - F - Me - F - Me - F - F - Me - Me - Me - Me - F - Me - F - Me - Me -Me - Me - Me - Me - Me; Alternatively, modify mode 3: Second link (5'-3'): Me - Me - Me - Me - Me - Me - F- Me - F- F - F - F - Me - Me - Me - Me - Me - Me - Me - Me - Me – Me, First link (5'-3'): Me -F - Me - F - Me - F - Me - F - F - Me - F - Me - Me - F - Me - F - Me - Me - Me - Me - Me – Me; Alternatively, modify mode 4: Second link (5'-3'): Me - Me - Me - Me - Me - Me - F - Me -F - F - F - F - Me - Me - Me - Me - Me - Me - Me - Me - Me - Me - Me, First link (5'-3'): Me- F - Me - F - Me - F - Me - Me - Me - Me - Me - Me - Me - Me - Me - Me - Me - Me; Alternatively, modify mode 5: Second link (5'-3') Traditional Me - Me - Me - Me - Me - Me - Me - Me - F - F - F - Me - Me - Me - Me - Me - Me - Me - Me - Me - Me - Me - Me, First link (5'-3') Traditional me - F - Me - Me - Me - F - Me - F - Me - Me - F - Me - Me - F - Me - Me - Me - Me - Me - Me; Alternatively, modify mode 6: Second link (5'-3' Traditional Me - Me - Me - Me - Me - Me - F - Me -F - F - F - Me - Me - Me - Me - Me - Me - Me - Me - Me - Me - Me - Me, First link (5'-3' Traditional me - F - Me - Me - Me - F - Me - F - Me - Me - F - Me - Me - Me - F - Me - Me - Me - Me - Me - Me.

[0444] One kind of nucleic acid, among them, one kind of modification model including the following: Modification pattern 1: Second pin (5'-3'): Me(s)Me(s)Me - Me - Me - Me - F - F - F - F- F - Me - Me - Me - Me - Me - Me - Me - Me - F - Me – Me, First pin (5'-3'): Me(s)F(s)Me - F - Me - F - Me - F - F - Me - Me - Me - Me - F - Me - F - Me - Me -Me - Me - Me(s)Me(s)Me; Some modification model 2: Second pin (5'-3'): Me(s)Me(s)Me - Me - Me - F - F - Me - F- F - F - F - Me - Me - Me - Me - Me - Me - Me - Me – Me, First pin (5'-3'): Me(s)F(s)Me - F - Me - F - Me - F - F - Me - Me - Me - Me - F - Me - F - Me -Me - Me - Me - Me(s)Me(s)Me; Some modification model 3: Second pin (5'-3'): Me(s)Me(s)Me - Me - Me - Me -F- Me - F- F - F - F - Me - Me - Me - Me - Me - Me - Me - Me – Me, First pin (5'-3'): Me(s)F(s)Me - F - Me - F - Me - F - F - Me - Me - Me - Me - F - Me - F - Me -Me - Me - Me - Me(s)Me(s)Me; Or modification pattern 4: Second strand (5’-3’): Me(s)Me(s)Me - Me - Me - Me - Me - F - Me -F - F - F - F - Me - Me - Me - Me - Me - Me - Me - Me – Me, First strand (5’-3’): Me(s)F(s)Me - F - Me - F - Me - Me - Me - Me - Me - Me - Me - F - Me - F - Me -Me - Me - Me - Me(s)Me(s)Me; Or modification pattern 5: Second strand (5’-3’): Me(s)Me(s)Me - Me - Me - Me - Me - Me -F - F - F - Me - Me - Me - Me - Me - Me - Me - Me - Me – Me, First strand (5’-3’):Me(s)F(s)Me - Me - Me - F - Me - Me - F - Me - Me - Me - Me - F - Me - F - Me- Me - Me - Me - Me(s)Me(s)Me; Or modification pattern 6: Second strand (5’-3’): Me(s)Me(s)Me - Me - Me - Me - F - Me -F - F - F - Me - Me - Me - Me - Me - Me - Me - Me - Me – Me, First strand (5’-3’):Me(s)F(s)Me - Me - Me - F - Me - Me - F - Me - Me - Me - Me - F - Me - F - Me- Me - Me - Me - Me(s)Me(s)Me; Wherein, (s) is a phosphorothioate internucleoside bond.

[0445] A nucleic acid, wherein the modified nucleosides include any one of the following modification patterns: Modification pattern 1: Second strand (5’-3’): Me – Me - Me - Me - Me - Me - F - F - F - F- F - Me - Me - Me - Me - Me - Me - Me - F(s)Me(s)Me, First strand (5’-3’): Me(s)F(s)Me - F - Me - F - Me - F - F - Me - Me - Me - Me - F - Me - F - Me - Me -Me - Me - Me - Me(s)Me(s)Me; Or modification pattern 2: Second strand (5’-3’): Me – Me - Me - Me - Me - F - F - Me - F- F - F - F - Me - Me - Me - Me - Me - Me - Me(s)Me(s)Me, First strand (5’-3’): Me(s)F(s)Me - F - Me - F - Me - F - F - Me - Me - Me - Me - F - Me - F - Me - Me - Me - Me - Me(s)Me(s)Me; Or modification pattern 3: Second strand (5’-3’): Me – Me - Me - Me - Me - Me -F- Me - F- F - F - F - Me - Me - Me - Me - Me - Me - Me(s)Me(s)Me, First strand (5’-3’): Me(s)F(s)Me - F - Me - F - Me - F - F - Me - Me - Me - Me - F - Me - F - Me - Me - Me - Me - Me(s)Me(s)Me; Or modification pattern 4: Second strand (5’-3’): Me – Me - Me - Me - Me - Me - F - Me -F - F - F - F - Me - Me - Me - Me - Me - Me - Me(s)Me(s)Me, First strand (5’-3’): Me(s)F(s)Me - F - Me - F - Me - Me - Me - Me - Me - Me - Me - F - Me - F - Me - Me - Me - Me - Me(s)Me(s)Me; Or modification pattern 5: Second strand (5’-3’): Me – Me - Me - Me - Me - Me - Me - Me -F - F - F - Me - Me - Me - Me - Me - Me - Me - Me(s)Me(s)Me, First strand (5’-3’):Me(s)F(s)Me - Me - Me - F - Me - Me - F - Me - Me - Me - Me - F - Me - F - Me- Me - Me - Me - Me(s)Me(s)Me; Or modification pattern 6: Second strand (5’-3’): Me – Me - Me - Me - Me - Me - F - Me -F - F - F - Me - Me - Me - Me - Me - Me - Me - Me(s)Me(s)Me, First strand (5’-3’):Me(s)F(s)Me - Me - Me - F - Me - Me - F - Me - Me - Me - Me - F - Me - F - Me- Me - Me - Me - Me(s)Me(s)Me; where (s) is a phosphorothioate internucleoside bond.

[0446] A nucleic acid, wherein the modified nucleosides include any one of the following modification patterns: Modification pattern 1: Second strand (5’-3’): ia – ia - Me - Me - Me - Me - Me - Me - F -F - F - F - F - Me - Me - Me - Me - Me - Me - Me - F - Me – Me, First strand (5’-3’): Me - F - Me - F - Me - F - Me - F - F - Me - Me - Me - Me - F - Me - F -Me - Me - Me - Me - Me - Me – Me; Alternatively, modify mode 2: Second link (5'-3'): ia – ia - Me - Me - Me - Me - Me - F -F - Me - F - F - F - F - Me - Me - Me - Me - Me - Me - Me - Me - Me, First link (5'-3'): Me - F - Me - F - Me - F - Me - F - F - Me - F - Me - Me - F - Me - F - Me - Me - Me - Me - Me - Me - Me; Alternatively, modify mode 3: Second link (5'-3'): ia – ia - Me - Me - Me - Me - Me - Me - F- Me - F - F - F - F - Me - Me - Me - Me - Me - Me - Me - Me - Me, First link (5'-3'): Me - F - Me - F - Me - F - Me - F - F - Me - F - Me - Me - F - Me - F - Me - Me - Me - Me - Me - Me; Alternatively, modify mode 4: Second link (5'-3'): ia – ia - Me - Me - Me - Me - Me - Me - F - Me - F - F - F - F - Me - Me - Me - Me - Me - Me - Me - Me - Me, First link (5'-3'): Me - F - Me - F - Me - F - Me - Me - Me - Me - Me - Me - Me - Me - F - Me- F - Me - Me - Me - Me - Me - Me - Me; Alternatively, modify mode 5: Second link (5'-3') Traditional ia – ia - Me - Me - Me - Me - Me - Me - Me - Me - Me - F - F - F - Me - Me - Me - Me - Me - Me - Me - Me - Me - Me, First link (5'-3') Traditional Me - F - Me - Me - Me - F - Me - F - Me - Me - F - Me - Me - Me - Me - F - Me- F - Me - Me - Me - Me - Me - Me - Me; Alternatively, modify mode 6: Second link (5'-3'): ia – ia - Me - Me - Me - Me - Me - Me - F - Me - F - F - F - Me - Me - Me - Me - Me - Me - Me - Me - Me - Me, First link (5'-3'): Me - F - Me - Me - Me - F - Me - F - Me - Me - F - Me - Me - Me - Me - F - Me- F - Me - Me - Me - Me - Me - Me - Me,; Among them, the ia display has no opposite direction.

[0447] One kind of nucleic acid, among them, one kind of modification model including the following: Modification model 1: Second pin (5'-3'): Me - Me - Me - Me - Me - Me - F - F - F - F- F - Me - Me - Me - Me - Me - Me - Me - Me - F - Me – Me - ia – ia, First pin (5'-3'): Me - F - Me - F - Me - F - Me - F - F - Me - Me - Me - Me - F - Me - F -Me - Me - Me - Me - Me - Me – Me; Alternatively, modify mode 2: Second link (5'-3'): Me - Me - Me - Me - Me - F - F - Me - F- F - F - F - Me - Me - Me - Me - Me - Me - Me - Me - Me - Me - Me - Me - ia - ia, First link (5'-3'): Me - F - Me - F - Me - F - Me - F - F - Me - F - Me - Me - F - Me - Me - Me - Me - Me - Me - Me - Me - Me; Alternatively, modify mode 3: Second link (5'-3'): Me - Me - Me - Me - Me - Me - F- Me - F- F - F - F - Me - Me - Me - Me - Me - Me - Me - Me - Me - Me - Me - Me - Me - Me - Me - Me - Me - Me - Me - Me - Me - Me; Alternatively, modify mode 4: Second link (5'-3'): Me - Me - Me - Me - Me - Me - F - Me -F - F - F - F - Me - Me - Me - Me - Me - Me - Me - Me - Me - Me - Me - Me - Me - Me - Me - Me - Me - Me - Me - Me - Me; First link (5'-3'): Me - F - Me - F - Me - F - Me - F - Me - Me - Me - Me - Me - Me - Me - Me - Me - Me; Or modification mode 5: Second strand (5'-3'): Me - Me - Me - Me - Me - Me - Me - Me -F - F - F - Me - Me - Me - Me - Me - Me - Me - Me - Me - Me - ia - ia, First strand (5'-3'): Me - F - Me - Me - Me - F - Me - Me - F - Me - Me - Me - Me - F - Me- F - Me - Me - Me - Me - Me - Me - Me; Or modification mode 6: Second strand (5'-3'): Me - Me - Me - Me - Me - Me - F - Me -F - F - F - Me - Me - Me - Me - Me - Me - Me - Me - Me - Me - ia - ia, - First strand (5'-3'): Me - F - Me - Me - Me - F - Me - Me - F - Me - Me - Me - Me - F -Me - F - Me - Me - Me - Me - Me - Me –Me,; Wherein, ia represents a reverse abasic nucleoside, and when the reverse abasic nucleoside represented by ia-ia is present at the 3' end of the second chain, the reverse abasic nucleoside exists as two nucleosides with protruding ends.

[0448] A nucleic acid wherein the modified nucleoside includes any of the following modification patterns: Modification pattern 1: Second strand (5'-3'): ia – ia - Me(s)Me(s)Me - Me - Me - Me - F -F - F - F - F - Me - Me - Me - Me - Me - Me - F - Me - Me, First strand (5'-3'): Me(s)F(s)Me - F - Me - F - Me - F - F - Me - Me - Me - Me - F - Me - F -Me - Me - Me - Me - Me(s)Me(s)Me; Or modification pattern 2: Second strand (5’-3’): ia – ia - Me(s)Me(s)Me - Me - Me - Me - F - F - Me - F - F - F - F - Me - Me - Me - Me - Me - Me - Me - Me - Me, First strand (5’-3’): Me(s)F(s)Me - F - Me - F - Me - F - F - Me - Me - Me - Me - F - Me - F - Me - Me - Me - Me - Me(s)Me(s)Me; Or modification pattern 3: Second strand (5’-3’): ia – ia - Me(s)Me(s)Me - Me - Me - Me - F - Me - F - F - F - F - Me - Me - Me - Me - Me - Me - Me - Me - Me, First strand (5’-3’): Me(s)F(s)Me - F - Me - F - Me - F - F - Me - Me - Me - Me - F - Me - F - Me - Me - Me - Me - Me(s)Me(s)Me; Or modification pattern 4: Second strand (5’-3’): ia – ia - Me(s)Me(s)Me - Me - Me - Me - F - Me - F - F - F - F - Me - Me - Me - Me - Me - Me - Me - Me – Me, First strand (5’-3’): Me(s)F(s)Me - F - Me - F - Me - Me - Me - Me - Me - Me - Me - F - Me - F - Me - Me - Me - Me - Me(s)Me(s)Me; Or modification pattern 5: Second strand (5’-3’): ia – ia - Me(s)Me(s)Me - Me - Me - Me - Me - Me - Me - F - F - F - Me - Me - Me - Me - Me - Me - Me - Me - Me - Me, First strand (5’-3’): Me(s)F(s)Me - Me - Me - F - Me - Me - F - Me - Me - Me - Me - F - Me - F - Me - Me - Me - Me - Me(s)Me(s)Me; Or modification pattern 6: Second strand (5’-3’): ia – ia - Me(s)Me(s)Me - Me - Me - Me - F - Me - F - F - F - Me - Me - Me - Me - Me - Me - Me - Me - Me - Me, First strand (5’-3’): Me(s)F(s)Me - Me - Me - F - Me - Me - F - Me - Me - Me - Me - F - Me - F - Me - Me - Me - Me - Me(s)Me(s)Me; Where: (s) represents a phosphorothioate internucleoside bond, and ia represents an inverted abasic nucleoside.

[0449] A nucleic acid, wherein the modified nucleoside includes any one of the following modification patterns: Modification pattern 1: Second strand (5’-3’): Me – Me - Me - Me - Me - Me - F - F - F - F - F - Me - Me - Me - Me - Me - Me - Me - F(s)Me(s)Me - ia – ia, First strand (5’-3’): Me(s)F(s)Me - F - Me - F - Me - F - F - Me - Me - Me - Me - F - Me - F - Me - Me - Me - Me - Me(s)Me(s)Me; Or modification pattern 2: Second strand (5’-3’): Me – Me - Me - Me - Me - F - F - Me - F- F - F - F - Me - Me - Me - Me - Me - Me - Me(s)Me(s)Me - ia – ia, First strand (5’-3’): Me(s)F(s)Me - F - Me - F - Me - F - F - Me - Me - Me - Me - F - Me -F - Me - Me - Me - Me - Me(s)Me(s)Me; Or modification pattern 3: Second strand (5’-3’): Me – Me - Me - Me - Me - Me -F- Me - F- F - F - F - Me - Me - Me - Me - Me - Me - Me(s)Me(s)Me - ia – ia, First strand (5’-3’): Me(s)F(s)Me - F - Me - F - Me - F - F - Me - Me - Me - Me - F - Me -F - Me - Me - Me - Me - Me(s)Me(s)Me; Or modification pattern 4: Second strand (5’-3’): Me – Me - Me - Me - Me - Me - F - Me -F - F - F - F - Me - Me - Me - Me - Me - Me - Me(s)Me(s)Me - ia – ia, First strand (5’-3’): Me(s)F(s)Me - F - Me - F - Me - Me - Me - Me - Me - Me - Me - F - Me- F - Me - Me - Me - Me - Me(s)Me(s)Me; Or modification mode 5: Second strand (5'-3'): Me - Me - Me - Me - Me - Me - Me - Me -F - F - F - Me - Me - Me - Me - Me - Me - Me - Me(s)Me(s)Me - ia - ia, First strand (5'-3'): Me(s)F(s)Me - Me - Me - F - Me - Me - F - Me - Me - Me - Me - F - Me- F - Me - Me - Me - Me - Me(s)Me(s)Me; Or modification pattern 6: Second strand (5'-3'): Me - Me - Me - Me - Me - Me - F - Me -F - F - F - Me - Me - Me - Me - Me - Me - Me - Me(s)Me(s)Me - ia - ia, First strand (5'-3'): Me(s)F(s)Me - Me - Me - F - Me - Me - F - Me - Me - Me - Me - F - Me- F - Me - Me - Me - Me - Me(s)Me(s)Me; Wherein, (s) is a thiophosphate nucleoside bond, ia represents a reverse abasic nucleoside, and when the reverse abasic nucleoside represented by ia-ia is present at the 3' end of the second chain, the reverse abasic nucleoside exists as two nucleoside overhangs.

[0450] Nucleic acid, wherein the first strand contains a 2' sugar modification pattern, wherein the modification is selected from at least 2'Me sugar modification and 2'F sugar modification, provided that the total number of 2'F sugar modifications in the first strand is not four or six.

[0451] Nucleic acid, wherein the first strand contains a 2' sugar modification pattern, wherein the modification is selected from at least 2'Me sugar modification and 2'F sugar modification, wherein the total number of 2'F sugar modifications in the first strand is three, five or seven.

[0452] Nucleic acid, wherein the first strand contains a 2' sugar modification pattern, wherein the modification is selected from at least 2'Me sugar modification and 2'F sugar modification, wherein the total number of 2'F sugar modifications in the first strand is three.

[0453] Nucleic acid, wherein the first strand contains a 2' sugar modification pattern, wherein the modification is selected from at least 2'Me sugar modification and 2'F sugar modification, wherein the total number of 2'F sugar modifications in the first strand is five.

[0454] Nucleic acid, wherein the first strand includes the following 2' sugar modification pattern (5'-3'): Me – F – Me – X2– Me – F – (Me)7– (F – Me)2– X3– Me – X4– (Me)3; X2, X3, and X4 are selected from 2'Me and 2'F sugar modifications, but for X2, X3, and X4, at least one is a 2'F sugar modification and the other two are 2'Me sugar modifications.

[0455] Nucleic acid, wherein the first strand includes the following 2' sugar modification pattern (5'-3'): Me – F – Me – X2– Me – F – (Me)7– (F – Me)2– X3– Me – X4– (Me)3; X2 is a 2'F sugar modification, while X3 and X4 are 2'Me sugar modifications.

[0456] Nucleic acid, wherein the first strand includes the following 2' sugar modification pattern (5'-3'): Me – F – Me – X2– Me – F – (Me)7– (F – Me)2– X3– Me – X4– (Me)3; X3 is a 2'F sugar modification, while X2 and X4 are 2'Me sugar modifications.

[0457] Nucleic acid, wherein the first strand includes the following 2' sugar modification pattern (5'-3'): Me – F – Me – X2– Me – F – (Me)7– (F – Me)2– X3– Me – X4– (Me)3; X4 is a 2'F sugar modification, while X2 and X3 are 2'Me sugar modifications.

[0458] Nucleic acid, wherein the first strand includes a 2' sugar modification pattern, the modification being selected from at least 2'Me sugar modification and 2'F sugar modification, wherein the total number of 2'F sugar modifications in the first strand is seven 2'F modifications.

[0459] Nucleic acid, wherein the first strand includes the following 2' sugar modification pattern (5'-3'): Me–F–Me–X2–Me–F–Me–(F)2–(Me)4–(F–Me)2–X3–Me–X4–(Me)3; X2, X3, and X4 are selected from 2'Me and 2'F sugar modifications, provided that for X2, X3, and X4, at least one is a 2'F sugar modification and the other two are 2'Me sugar modifications.

[0460] Nucleic acid, wherein the first strand includes the following 2' sugar modification pattern (5'-3'): Me–F–Me–X2–Me–F–Me–(F)2–(Me)4–(F–Me)2–X3–Me–X4–(Me)3; X2 is a 2'F sugar modification, while X3 and X4 are 2'Me sugar modifications.

[0461] Nucleic acid, wherein the first strand includes the following 2' sugar modification pattern (5'-3'): Me–F–Me–X2–Me–F–Me–(F)2–(Me)4–(F–Me)2–X3–Me–X4–(Me)3; X3 is a 2'F sugar modification, while X2 and X4 are 2'Me sugar modifications.

[0462] Nucleic acid, wherein the first strand includes the following 2' sugar modification pattern (5'-3'): Me–F–Me–X2–Me–F–Me–(F)2–(Me)4–(F–Me)2–X3–Me–X4–(Me)3; X4 is a 2'F sugar modification, while X2 and X3 are 2'Me sugar modifications.

[0463] Nucleic acid, wherein the first strand includes the following 2' sugar modification pattern (5'-3'): Me – F – (Me)3– X1– (Me)7– F – Me – F – (Me)7; where X1 is a thermally unstable modification.

[0464] Nucleic acid, wherein the first strand includes the following 2' sugar modification pattern (5'-3'): Me – F – (Me)3– X1– Me – (F)2– (Me)4– F – Me – F – (Me)7; where X1 is a thermally unstable modification.

[0465] Nucleic acid, wherein the second strand includes the following 2' sugar modification pattern (5'-3'): (Me)8–(F)3–(Me) 10 .

[0466] Nucleic acid, wherein the second strand includes the following 2' sugar modification pattern (5'-3'): (Me)8– (F)3– (Me) 10 ,and The first chain includes a 2' sugar modification pattern, wherein the modification is selected from at least 2'Me sugar modification and 2'F sugar modification, provided that the total number of 2'F sugar modifications in the first chain is not four or six 2'F modifications.

[0467] Nucleic acid, wherein the second strand includes the following 2' sugar modification pattern (5'-3'): (Me)8– (F)3– (Me) 10 ,and The first chain includes a 2' sugar modification pattern, wherein the modification is selected from at least 2'Me and 2'F sugar modifications, wherein the total number of 2'F sugar modifications in the first chain is three, five, or seven.

[0468] Nucleic acid, comprising a first strand at least partially complementary to a portion of RNA transcribed from a target gene, and a second strand at least partially complementary to the first strand, wherein the first and second strands form a double-stranded region of at least 17 nucleotides in length, and wherein the nucleotides of the second strand comprise the following 2' sugar modification pattern (5'-3'): (Me)8– (F)3– (Me) 10 ,and The nucleoside of the first chain includes the following 2' sugar modification patterns (5'-3'): Me – F – (Me)3– X1– (Me)7– F – Me – F – (Me)7, where X1 is a thermally unstable modification.

[0469] Nucleic acid, comprising a first strand at least partially complementary to a portion of RNA transcribed from a target gene, and a second strand at least partially complementary to the first strand, wherein the first and second strands form a double-stranded region of at least 17 nucleotides in length, and wherein the nucleotides of the second strand comprise the following 2' sugar modification pattern (5'-3'): (Me)8–(F)3–(Me). 10 ,and The nucleosides of the first chain include the following 2' sugar modification patterns (5'-3'): Me – F – (Me)3 – X1 – (Me)7 – F – Me – F – (Me)7.

[0470] Nucleic acid, comprising a first strand at least partially complementary to a portion of RNA transcribed from a target gene, and a second strand at least partially complementary to the first strand, wherein the first and second strands form a double-stranded region of at least 17 nucleotides in length, and wherein the nucleotides of the second strand comprise the following 2' sugar modification pattern (5'-3'): (Me)8–(F)3–(Me). 10 ,and The nucleosides of the first chain include the following 2' sugar modification patterns (5'-3'): Me – F – (Me)3– F – (Me)7– (F – Me)2– F – (Me)5.

[0471] Nucleic acid, comprising a first strand at least partially complementary to a portion of RNA transcribed from a target gene, and a second strand at least partially complementary to the first strand, wherein the first and second strands form a double-stranded region of at least 17 nucleotides in length, and wherein the nucleotides of the second strand comprise the following 2' sugar modification pattern (5'-3'): (Me)8–(F)3–(Me). 10 ,and The nucleosides of the first chain include the following 2' sugar modification patterns (5'-3'): Me – F – (Me)3– F – (Me)7– F – Me – F – (Me)3– F – (Me)3.

[0472] Nucleic acid, comprising a first strand at least partially complementary to a portion of RNA transcribed from a target gene, and a second strand at least partially complementary to the first strand, wherein the first and second strands form a double-stranded region of at least 17 nucleotides in length, and wherein the nucleotides of the second strand comprise the following 2' sugar modification pattern (5'-3'): (Me)8–(F)3–(Me). 10 ,and The nucleosides of the first chain include the following 2' sugar modification patterns (5'-3'): Me – F – (Me)3– X1– Me – (F)2– (Me)4– F – Me – F – (Me)7, where X1 is a thermally unstable modification.

[0473] Nucleic acid, comprising a first strand at least partially complementary to a portion of RNA transcribed from a target gene, and a second strand at least partially complementary to the first strand, wherein the first and second strands form a double-stranded region of at least 17 nucleotides in length, and wherein the nucleotides of the second strand comprise the following 2' sugar modification pattern (5'-3'): (Me)8–(F)3–(Me). 10 ,and The nucleosides of the first chain include the following 2' sugar modification patterns (5'-3'): (Me – F)3– Me – (F)2– (Me)4– (F – Me)2– (Me)6.

[0474] Nucleic acid, comprising a first strand at least partially complementary to a portion of RNA transcribed from a target gene, and a second strand at least partially complementary to the first strand, wherein the first and second strands form a double-stranded region of at least 17 nucleotides in length, and wherein the nucleotides of the second strand comprise the following 2' sugar modification pattern (5'-3'): (Me)8–(F)3–(Me). 10,and The nucleosides of the first chain include the following 2' sugar modification patterns (5'-3'): Me – F – (Me)3– F – Me – (F)2– (Me)4– (F – Me)2– F – (Me)5.

[0475] Nucleic acid, comprising a first strand at least partially complementary to a portion of RNA transcribed from a target gene, and a second strand at least partially complementary to the first strand, wherein the first and second strands form a double-stranded region of at least 17 nucleotides in length, and wherein the nucleotides of the second strand comprise the following 2' sugar modification pattern (5'-3'): (Me)8–(F)3–(Me). 10 ,and The nucleosides of the first chain include the following 2' sugar modification patterns (5'-3'): Me – F – (Me)3– F – Me – (F)2– (Me)4– (F – Me)2– (Me)2– F – (Me)3.

[0476] Nucleic acid, wherein the second strand includes a 2' sugar and the following base-free modification pattern (5'-3'): ia-ia-(Me)8–(F)3–(Me) 10 ; where ia represents a reverse abase-free nucleoside.

[0477] Nucleic acid, wherein the second strand includes a 2' sugar and the following base-free modification pattern (5'-3'): ia-ia-(Me)8– (F)3– (Me) 10 Where ia represents a reverse anucleotide, and The first chain includes a 2' sugar modification pattern, wherein the modification is selected from at least 2'Me sugar modification and 2'F sugar modification, provided that the total number of 2'F sugar modifications in the first chain is not four or six 2'F modifications.

[0478] Nucleic acid, wherein the second strand includes a 2' sugar and the following base-free modification pattern (5'-3'): ia-ia-(Me)8 – (F)3 – (Me)10, where ia represents a reverse abase-free nucleoside, and The first chain includes a 2' sugar modification pattern, wherein the modification is selected from at least 2'Me sugar modification and 2'F sugar modification, and the total number of 2'F sugar modifications in the first chain is three, five or seven 2'F modifications.

[0479] Nucleic acid, comprising a first strand at least partially complementary to a portion of RNA transcribed from a target gene, and a second strand at least partially complementary to the first strand, wherein the first and second strands form a double-stranded region of at least 17 nucleotides in length, and wherein the nucleotides of the second strand comprise a 2' sugar, and the following base-free modification pattern (5'-3'): ia-ia-(Me)8– (F)3– (Me) 10 Where ia represents a reverse anucleotide, and The nucleosides of the first chain include the following 2' sugar modification patterns (5'-3'): Me – F – (Me)3– X1– (Me)7– F – Me – F – (Me)7, where X1 is a thermally unstable modification.

[0480] Nucleic acid, comprising a first strand at least partially complementary to a portion of RNA transcribed from a target gene, and a second strand at least partially complementary to the first strand, wherein the first and second strands form a double-stranded region of at least 17 nucleotides in length, and wherein the nucleotides of the second strand comprise a 2' sugar, and the following base-free modification pattern (5'-3'): ia-ia-(Me)8– (F)3– (Me) 10 Where ia represents a reverse anucleotide, and The nucleosides of the first chain include the following 2' sugar modification patterns (5'-3'): (Me – F)3– (Me)7– F – Me – F – (Me)7.

[0481] Nucleic acid, comprising a first strand at least partially complementary to a portion of RNA transcribed from a target gene, and a second strand at least partially complementary to the first strand, wherein the first and second strands form a double-stranded region of at least 17 nucleotides in length, and wherein the nucleotides of the second strand comprise a 2' sugar, and the following base-free modification pattern (5'-3'): ia-ia-(Me)8– (F)3– (Me) 10 Where ia represents a reverse anucleotide, and The nucleosides of the first chain include the following 2' sugar modification patterns (5'-3'): Me – F – (Me)3– F – (Me)7– (F – Me)2– F – (Me)5.

[0482] Nucleic acid, comprising a first strand at least partially complementary to a portion of RNA transcribed from a target gene, and a second strand at least partially complementary to the first strand, wherein the first and second strands form a double-stranded region of at least 17 nucleotides in length, and wherein the nucleotides of the second strand comprise a 2' sugar, and the following base-free modification pattern (5'-3'): ia-ia-(Me)8– (F)3– (Me) 10 Where ia represents a reverse anucleotide, and The nucleosides of the first chain include the following 2' sugar modification patterns (5'-3'): Me – F – (Me)3– F – (Me)7– F – Me – F – (Me)3– F – (Me)3.

[0483] Nucleic acid, comprising a first strand at least partially complementary to a portion of RNA transcribed from a target gene, and a second strand at least partially complementary to the first strand, wherein the first and second strands form a double-stranded region of at least 17 nucleotides in length, and wherein the nucleotides of the second strand comprise a 2' sugar, and the following base-free modification pattern (5'-3'): ia-ia-(Me)8– (F)3– (Me) 10 Where ia represents a reverse anucleotide, and The nucleosides of the first chain include the following 2' sugar modification patterns (5'-3'): Me – F – (Me)3– X1– Me – (F)2– (Me)4– F – Me – F – (Me)7, X1 is a thermally unstable modification.

[0484] Nucleic acid, comprising a first strand at least partially complementary to a portion of RNA transcribed from a target gene, and a second strand at least partially complementary to the first strand, wherein the first and second strands form a double-stranded region of at least 17 nucleotides in length, and wherein the nucleotides of the second strand comprise a 2' sugar, and the following base-free modification pattern (5'-3'): ia-ia-(Me)8– (F)3– (Me) 10 Where ia represents a reverse anucleotide, and The nucleosides of the first chain include the following 2' sugar modification patterns (5'-3'): (Me – F)3– Me – (F)2– (Me)4– (F – Me)2– (Me)6.

[0485] Nucleic acid, comprising a first strand at least partially complementary to a portion of RNA transcribed from a target gene, and a second strand at least partially complementary to the first strand, wherein the first and second strands form a double-stranded region of at least 17 nucleotides in length, and wherein the nucleotides of the second strand comprise a 2' sugar, and the following base-free modification pattern (5'-3'): ia-ia-(Me)8– (F)3– (Me) 10 Where ia represents a reverse anucleotide, and The nucleosides of the first chain include the following 2' sugar modification patterns (5'-3'): Me – F – (Me)3– F – Me – (F)2– (Me)4– (F – Me)2– F – (Me)5.

[0486] Nucleic acid, comprising a first strand at least partially complementary to a portion of RNA transcribed from a target gene, and a second strand at least partially complementary to the first strand, wherein the first and second strands form a double-stranded region of at least 17 nucleotides in length, and wherein the nucleotides of the second strand comprise a 2' sugar, and the following base-free modification pattern (5'-3'): ia-ia-(Me)8– (F)3– (Me) 10 Where ia represents a reverse anucleotide, and The nucleosides of the first chain include the following 2' sugar modification patterns (5'-3'): Me – F – (Me)3– F – Me – (F)2– (Me)4– (F – Me)2– (Me)2– F – (Me)3.

[0487] Nucleic acid, wherein the second strand includes the following 2' sugar modification pattern (5'-3'): ia-ia-Me(s)Me(s) (Me)6– (F)3– (Me) 10 ; Where ia represents a reverse abasic nucleoside, and (s) represents a thiophosphate nucleoside bond.

[0488] Nucleic acid, wherein the second strand includes the following 2' sugar modification pattern (5'-3'): ia-ia-Me(s)Me(s) (Me)6– (F)3– (Me) 10 Where ia represents a reverse abasic nucleoside, and (s) represents a thiophosphate nucleoside internucleotide bond; and The first chain includes a 2' sugar modification pattern, wherein the modification is selected from at least 2'Me sugar modification and 2'F sugar modification, provided that the total number of 2'F sugar modifications in the first chain is not four or six 2'F modifications.

[0489] Nucleic acid, wherein the second strand includes the following 2' sugar modification pattern (5'-3'): ia-ia-Me(s)Me(s) (Me)6– (F)3– (Me) 10 Where ia represents a reverse abasic nucleoside, and (s) represents a thiophosphate nucleoside internucleotide bond; and The first chain includes a 2' sugar modification pattern, wherein the modification is selected from at least 2'Me sugar modification and 2'F sugar modification, but the total number of 2'F sugar modifications in the first chain is three, five or seven 2'F modifications.

[0490] Nucleic acid, comprising a first strand at least partially complementary to a portion of RNA transcribed from a target gene, and a second strand at least partially complementary to the first strand, wherein the first and second strands form a double-stranded region of at least 17 nucleotides in length, and wherein the nucleotides of the second strand comprise a 2' sugar, and the following base-free modification pattern (5'-3'): ia-ia-Me(s)Me(s) (Me)6– (F)3– (Me) 10 Where ia represents a reverse abasic nucleoside, and (s) represents a thiophosphate nucleoside internucleotide bond; and The nucleosides of the first chain include the following 2' sugar modification patterns (5'-3'): Me(s)F(s)(Me)3– X1– (Me)7– F – Me – F – (Me)5(s)Me(s)Me, where X1 is a thermally unstable modification.

[0491] Nucleic acid, comprising a first strand at least partially complementary to a portion of RNA transcribed from a target gene, and a second strand at least partially complementary to the first strand, wherein the first and second strands form a double-stranded region of at least 17 nucleotides in length, and wherein the nucleotides of the second strand comprise a 2' sugar, and the following base-free modification pattern (5'-3'): ia-ia-Me(s)Me(s) (Me)6– (F)3– (Me) 10 Where ia represents a reverse abasic nucleoside, and (s) represents a thiophosphate nucleoside internucleotide bond; and The nucleosides of the first chain include the following 2' sugar modification patterns (5'-3'): Me(s)F(s)Me – F – Me – F – (Me)7– F – Me – F – (Me)5(s)Me(s)Me.

[0492] Nucleic acid, comprising a first strand at least partially complementary to a portion of RNA transcribed from a target gene, and a second strand at least partially complementary to the first strand, wherein the first and second strands form a double-stranded region of at least 17 nucleotides in length, and wherein the nucleotides of the second strand comprise a 2' sugar, and the following base-free modification pattern (5'-3'): ia-ia-Me(s)Me(s) (Me)6– (F)3– (Me) 10 Where ia represents a reverse abasic nucleoside, and (s) represents a thiophosphate nucleoside internucleotide bond; and The nucleosides of the first chain include the following 2' sugar modification patterns (5'-3'): Me(s)F(s)(Me)3– F – (Me)7– (F – Me)2– F – (Me)3(s)Me(s)Me.

[0493] Nucleic acid, comprising a first strand at least partially complementary to a portion of RNA transcribed from a target gene, and a second strand at least partially complementary to the first strand, wherein the first and second strands form a double-stranded region of at least 17 nucleotides in length, and wherein the nucleotides of the second strand comprise a 2' sugar, and the following base-free modification pattern (5'-3'): ia-ia-Me(s)Me(s) (Me)6– (F)3– (Me) 10 Where ia represents a reverse abasic nucleoside, and (s) represents a thiophosphate nucleoside internucleotide bond; and The nucleosides of the first chain include the following 2' sugar modification patterns (5'-3'): Me(s)F(s)(Me)3– F – (Me)7– F – Me – F – (Me)3– F – Me(s)Me(s)Me.

[0494] Nucleic acid, comprising a first strand at least partially complementary to a portion of RNA transcribed from a target gene, and a second strand at least partially complementary to the first strand, wherein the first and second strands form a double-stranded region of at least 17 nucleotides in length, and wherein the nucleotides of the second strand comprise a 2' sugar, and the following base-free modification pattern (5'-3'): ia-ia-Me(s)Me(s) (Me)6– (F)3– (Me) 10 Where ia represents a reverse abasic nucleoside, and (s) represents a thiophosphate nucleoside internucleotide bond; and The nucleosides of the first chain include the following 2' sugar modification patterns (5'-3'): Me(s)F(s)(Me)3– X1– Me – (F)2– (Me)4– F – Me – F – (Me)5(s)Me(s)Me, where X1 is a thermally unstable modification.

[0495] Nucleic acid, comprising a first strand at least partially complementary to a portion of RNA transcribed from a target gene, and a second strand at least partially complementary to the first strand, wherein the first and second strands form a double-stranded region of at least 17 nucleotides in length, and wherein the nucleotides of the second strand comprise a 2' sugar, and the following base-free modification pattern (5'-3'): ia-ia-Me(s)Me(s) (Me)6– (F)3– (Me) 10 Where ia represents a reverse abasic nucleoside, and (s) represents a thiophosphate nucleoside internucleotide bond; and The nucleosides of the first chain include the following 2' sugar modification patterns (5'-3'): Me(s)F(s)Me – F – Me – F – Me – (F)2– (Me)4– (F – Me)2– (Me)4(s)Me(s)Me.

[0496] Nucleic acid, comprising a first strand at least partially complementary to a portion of RNA transcribed from a target gene, and a second strand at least partially complementary to the first strand, wherein the first and second strands form a double-stranded region of at least 17 nucleotides in length, and wherein the nucleotides of the second strand comprise a 2' sugar, and the following base-free modification pattern (5'-3'): ia-ia-Me(s)Me(s) (Me)6– (F)3– (Me) 10 Where ia represents a reverse abasic nucleoside, and (s) represents a thiophosphate nucleoside internucleotide bond; and The nucleosides of the first chain include the following 2' sugar modification patterns (5'-3'): Me(s)F(s)(Me)3– F – Me – (F)2– (Me)4– (F – Me)2– F – (Me)3(s)Me(s)Me.

[0497] Nucleic acid, comprising a first strand at least partially complementary to a portion of RNA transcribed from a target gene, and a second strand at least partially complementary to the first strand, wherein the first and second strands form a double-stranded region of at least 17 nucleotides in length, and wherein the nucleotides of the second strand comprise a 2' sugar, and the following base-free modification pattern (5'-3'): ia-ia-Me(s)Me(s) (Me)6– (F)3– (Me) 10Where ia represents a reverse abasic nucleoside, and (s) represents a thiophosphate nucleoside internucleotide bond; and The nucleosides of the first chain include the following 2' sugar modification patterns (5'-3'): Me(s)F(s)(Me)3– F – Me – (F)2– (Me)4– (F – Me)2– (Me)2– F – Me(s)Me(s)Me.

[0498] The following modifications are preferred: Modification Mode 1: Second strand (5'-3'): ia – ia – Me – Me – Me – Me – Me – Me – Me – Me – F –F – F – Me – Me – Me – Me – Me – Me – Me – Me – Me – Me, First strand (5'-3'): Me – F – Me – Me – Me – X1– Me – Me – Me – Me – Me –Me – Me – F – Me – F – Me – Me – Me – Me – Me – Me – Me, where X1 is a thermally unstable modification; Or, modify mode 2: Second strand (5'-3'): ia – ia – Me – Me – Me – Me – Me – Me – Me – Me – F –F – F – Me – Me – Me – Me – Me – Me – Me – Me – Me – Me, First strand (5'-3'): Me – F – Me – F – Me – F – Me – Me – Me – Me – Me – Me – Me – F – Me – F – Me – Me – Me – Me – Me – Me – Me; Or modify mode 3: Second strand (5'-3'): ia – ia – Me – Me – Me – Me – Me – Me – Me – Me – F –F – F – Me – Me – Me – Me – Me – Me – Me – Me – Me – Me, First strand (5’-3’): Me – F – Me – Me – Me – F – Me – Me – Me – Me – Me – Me - Me – F – Me – F – Me – F – Me – Me – Me – Me - Me; Or modification pattern 4: Second strand (5’-3’): ia – ia – Me – Me – Me – Me – Me – Me – Me – Me – F – F – F – Me – Me – Me – Me – Me – Me – Me – Me – Me – Me, First strand (5’-3’): Me – F – Me – Me - Me – F – Me – Me – Me – Me – Me – Me - Me – F – Me – F – Me – Me – Me – F – Me – Me - Me; Or modification pattern 5: Second strand (5’-3’): ia – ia – Me – Me – Me – Me – Me – Me – Me – Me – F – F – F – Me – Me – Me – Me – Me – Me – Me – Me – Me – Me, First strand (5’-3’): Me – F – Me – Me - Me – X1– Me – F – F – Me – Me – Me – Me – F – Me – F – Me – Me – Me – Me – Me – Me – Me, where X1 is a thermally labile modification; Or modification pattern 6: Second strand (5’-3’): ia – ia – Me – Me – Me – Me – Me – Me – Me – Me – F – F – F – Me – Me – Me – Me – Me – Me – Me – Me – Me – Me, First strand (5’-3’): Me – F – Me – F – Me - F – Me – F – F – Me – Me – Me – Me – F – Me – F – Me – Me – Me – Me – Me – Me – Me; A certain model 7: Second pin (5'-3'): ia – ia – Me – Me – Me – Me – Me – Me – Me – Me – F –F – F – Me – Me – Me – Me – Me – Me – Me – Me – Me – Me, 1st pin (5'-3'): Me – F – Me – Me – Me – F – Me – F – F – Me– Me – Me –Me – F – Me – F – Me – F – Me – Me – Me – Me – Me; Model 8: Second pin (5'-3'): ia – ia – Me – Me – Me – Me – Me – Me – Me – Me – F –F – F – Me – Me – Me – Me – Me – Me – Me – Me – Me – Me, 1st pin (5'-3'): Me – F – Me – Me – Me – F – Me – F – F – Me – Me – Me – Me – F – Me – F – Me – Me – Me – F – Me – Me – Me. Specific selection below: Repair model 1: Second pin (5'-3'): ia – ia – Me(s)Me(s)Me – Me – Me – Me – Me – Me – F –F – F – Me – Me – Me – Me – Me – Me – Me – Me – Me – Me, First link (5'-3'): Me(s)F(s)Me – Me – Me – X1– Me – Me – Me – Me – Me – Me - Me – F – Me – F – Me – Me – Me – Me – Me(s)Me(s)Me, where X1 is the hot unsteady modification; A certain type of repair model 2: Second strand (5’-3’): ia – ia – Me(s)Me(s)Me – Me – Me – Me – Me – Me – F –F – F – Me – Me – Me – Me – Me – Me – Me – Me – Me – Me, First strand (5’-3’): Me(s)F(s)Me – F – Me – F – Me – Me – Me – Me – Me – Me– Me – F – Me – F – Me – Me – Me – Me – Me(s)Me(s)Me; Or modification pattern 3: Second strand (5’-3’): ia – ia – Me(s)Me(s)Me – Me – Me – Me – Me – Me – F –F – F – Me – Me – Me – Me – Me – Me – Me – Me – Me – Me, First strand (5’-3’): Me(s)F(s)Me – Me – Me – F – Me – Me – Me – Me – Me –Me - Me – F – Me – F – Me – F – Me – Me – Me(s)Me(s)Me; Or modification pattern 4: Second strand (5’-3’): ia – ia – Me(s)Me(s)Me – Me – Me – Me – Me – Me – F –F – F – Me – Me – Me – Me – Me – Me – Me – Me – Me – Me, First strand (5’-3’): Me(s)F(s)Me – Me - Me – F – Me – Me – Me – Me – Me –Me - Me – F – Me – F – Me – Me – Me – F – Me(s)Me(s)Me; Or modification pattern 5: Second strand (5’-3’): ia – ia – Me(s)Me(s)Me – Me – Me – Me – Me – Me – F –F – F – Me – Me – Me – Me – Me – Me – Me – Me – Me – Me, 1st pin (5'-3'): Me(s)F(s)Me – Me - Me – A certain modification model 6: Second pin (5'-3'): ia – ia – Me(s)Me(s)Me – Me – Me – Me – Me – Me – F –F – F – Me – Me – Me – Me – Me – Me – Me – Me – Me – Me, 1st pin (5'-3'): Me(s)F(s)Me – F – Me - F – Me – F – F – Me – Me – Me –Me – F – Me – F – Me – Me – Me – Me – Me(s)Me(s)Me; A certain model 7: Second pin (5'-3'): ia – ia – Me(s)Me(s)Me – Me – Me – Me – Me – Me – F –F – F – Me – Me – Me – Me – Me – Me – Me – Me – Me – Me, 1st pin (5'-3'): Me(s)F(s)Me – Me – Me – F – Me – F – F – Me– Me – Me –Me – F – Me – F – Me – F – Me – Me – Me(s)Me(s)Me; Model 8: Second pin (5'-3'): ia – ia – Me(s)Me(s)Me – Me – Me – Me – Me – Me – F –F – F – Me – Me – Me – Me – Me – Me – Me – Me – Me – Me, 1st pin (5'-3'): Me(s)F(s)Me – Me – Me – F – Me – F – F – Me – Me – Me– Me – F – Me – F – Me – Me – Me – F – Me(s)Me(s)Me; Wherein, (s) is the internucleotide bond between thiophosphate esters.

[0499] Nucleic acid conjugation with ligands Another modification of the nucleic acid (such as RNA, such as siRNA) in this application involves linking the nucleic acid (such as siRNA) to one or more ligand moieties, for example, to enhance the activity, cellular distribution, or cellular uptake of the nucleic acid (such as siRNA), such as entry into the cell.

[0500] In some embodiments, the ligand moiety can be linked to a nucleic acid, such as an siRNA oligonucleotide, via a cleavable or non-cleavable linker. The term "linker" or "connecting group" refers to an organic group that connects two parts of a compound, for example, covalently connecting the two parts of a compound.

[0501] The ligand can attach to the 3' or 5' end of the justice chain.

[0502] Preferably, the ligand is conjugated to the 3' end of the positive strand of a nucleic acid such as siRNA.

[0503] Therefore, this application further relates to conjugates for inhibiting the expression of targets (e.g., target genes) in cells, said conjugates comprising a nucleic acid moiety and one or more ligand moieties, said nucleic acid moiety comprising nucleic acids as disclosed herein.

[0504] On one hand, the second strand of the nucleic acid is directly or indirectly (e.g., through a linker) conjugated to one or more ligand moieties, wherein the ligand moieties are typically located in the terminal region of the second strand, preferably in its 3' terminal region.

[0505] In some embodiments, the ligand portion comprises GalNAc or a GalNAc derivative that is linked to a nucleic acid such as dsiRNA via a linker.

[0506] Therefore, this application relates to a conjugate wherein the ligand moiety comprises i) one or more GalNAc ligands; and / or ii) One or more GalNAc ligand derivatives; and / or iii) One or more GalNAc ligands conjugated to the nucleic acid via a linker.

[0507] The GalNAc ligand can be directly or indirectly conjugated to the 5' or 3' end region of the second strand of nucleic acid, preferably conjugated to its 3' end region.

[0508] GalNAc ligands are well known in the art, especially as described in EP3775207A1.

[0509] In some implementations, the ligand portion comprises one or more ligands.

[0510] In some implementations, the ligand portion comprises one or more carbohydrate ligands.

[0511] In some embodiments, the one or more carbohydrates may be monosaccharides, disaccharides, trisaccharides, tetrasaccharides, oligosaccharides, and / or polysaccharides.

[0512] In some embodiments, the one or more carbohydrates comprise one or more galactose moieties, one or more lactose moieties, one or more N-acetylgalactosamine moieties, and / or one or more mannose moieties.

[0513] In some embodiments, the one or more carbohydrates comprise one or more N-acetylgalactosamine moieties.

[0514] In some embodiments, the compounds described anywhere herein comprise two or three N-acetylgalactosamine moieties.

[0515] In some implementations, the one or more ligands are connected in a linear or branched configuration, for example, each configuration is connected to a branch point in the entire junction.

[0516] Exemplary linear and branching configurations are shown in Figures 1a and 1b: In Figure 1a (linear), (a) and / or (b) can typically represent linking bonds or linking groups, such as phosphate esters or thiophosphate ester groups.

[0517] In Figure 1b (branching), in some embodiments, the one or more ligands are connected in a biantennary or triantennary branching configuration. Typically, a triantennary branching configuration, such as the N-acetylgalactosamine triantennary branching configuration, is preferred.

[0518] connector The exemplary compounds of this application include a “connector portion”, as shown in Formula (I), which is part of the whole “connector”.

[0519] Formula (I) in: R1 is selected independently from hydrogen, methyl, and ethyl each time it appears; R2 is selected from the following group: hydrogen, hydroxyl group, -OC 1-3 Alkyl group, -C(=O)OC 1-3 Alkyl, halogen, and nitro groups; X1 and X2 are each selected independently from the following group: methylene, oxygen, and sulfur; m is an integer from 1 to 6; n is an integer from 1 to 10; q, r, s, t, v are independent integers from 0 to 4, provided that: (i) q and r cannot both be 0; and (ii) s, t and v cannot all be 0 at the same time; Z is the oligonucleotide moiety.

[0520] As further understood in the art, the exemplary compounds of this application comprise an entire connector located between the oligonucleotide moiety and the ligand moiety of these compounds. The entire connector “connects” the oligonucleotide moiety and the ligand moiety to each other.

[0521] The entire linker is generally assumed to contain one or more linker building blocks. For example, there exists a linker portion, described in Formula (I) as the “linker portion,” which is located adjacent to the ligand portion and typically connects the ligand portion directly or indirectly to the oligonucleotide portion via a branch point. The linker portion shown in Formula (I) is also often referred to as the “ligand arm” of the entire linker. Other linker portions may also exist between the oligonucleotide portion and the branch point (but not always), which are often referred to as the “tethering portion” of the entire linker, “tethering” the oligonucleotide portion to the remainder of the conjugated compound. Such 'ligand arms’ and / or 'linker portions’ and / or 'tethering portions’ can be understood with reference to the linear and / or branched configurations described above.

[0522] As can be seen from the claims and the remainder of the patent specification, the scope of this application covers both linear and branched configurations, and there is no limitation on the number of individual ligands that may be present. Furthermore, those skilled in the art, based on prior art and the expertise of oligonucleotide chemists, will recognize that many structures can be used as linker sites.

[0523] The remaining portion of the entire connector (excluding the connector portion) listed in the claims, as well as the remainder of the patent specification, has a chemical composition as shown in Formula (I), which the inventors consider to be particularly unique to this application. However, more generally, these chemical components can be described as the "tethered portion" as previously described, where the "tethered portion" is a part of the entire connector comprising the atomic group between Z (i.e., the oligonucleotide portion) and the connector portion, as shown in Formula (I).

[0524] The chain part of Formula I With respect to formula (I), the “chain portion” comprises the group of atoms between Z (i.e., the oligonucleotide portion) and the linker portion.

[0525] In some embodiments, R1 is always hydrogen. In some embodiments, R1 is methyl. In some embodiments, R1 is ethyl.

[0526] In some embodiments, R2 is a hydroxyl group. In some embodiments, R2 is a halogen. In some embodiments, R2 is fluorine. In some embodiments, R2 is chlorine. In some embodiments, R2 is bromine. In some embodiments, R2 is iodine. In some embodiments, R2 is a nitro group.

[0527] In some embodiments, X1 is methylene. In some embodiments, X1 is oxygen. In some embodiments, X1 is sulfur.

[0528] In some embodiments, X2 is methylene. In some embodiments, X2 is oxygen. In some embodiments, X2 is sulfur.

[0529] In some implementations, m=3.

[0530] In some implementations, n=6.

[0531] In some embodiments, X1 is oxygen and X2 is methylene. In some embodiments, both X1 and X2 are methylene.

[0532] In some implementations, q=1, r=2, s=1, t=1, v=1. In other implementations, q=1, r=3, s=1, t=1, v=1.

[0533] In some implementations, R1 is always hydrogen, n=6, m=3, R2 is fluorine, X2 is methylene, v=1, t=1, s=1, X1 is methylene, q=1 and r=2.

[0534] Therefore, in some embodiments, the exemplary compounds described in this application comprise the following structures: Formula (IV) In some implementations, R1 is hydrogen each time it appears, n=6, m=3, R2 is fluorine, X2 is methylene, v=1, t=1, s=1, X1 is oxygen, q=1 and r=2.

[0535] Therefore, in some embodiments, the exemplary compounds of this application comprise the following structures: Equation (II) Alternative chain part During the synthesis of the compounds in this application, alternative chain-linking structures may occur. In some embodiments, the alternative chain-linking portion has one or more atomic variations throughout the chain-linking portion of the connector compared to the chain-linking portion described anywhere herein.

[0536] In some embodiments, the alternative chain portion is a compound of formula (I) described anywhere herein, wherein R2 is a hydroxyl group.

[0537] In some implementations, R1 is hydrogen each time it appears, n=6, m=3, R2 is hydroxyl, X2 is methylene, v=1, t=1, s=1, X1 is methylene, q=1 and r=2.

[0538] Therefore, in some embodiments, the compound described in this application comprises the following structure: Formula (V) In some implementations, R1 is hydrogen each time it appears, n=6, m=3, R2 is hydroxyl, X2 is methylene, v=1, t=1, s=1, X1 is oxygen, q=1 and r=2.

[0539] Therefore, in some embodiments, the compound described in this application comprises the following structure: Equation (III) Connector section With regard to formula (I), the “connector portion” shown in formula (I) comprises the group of atoms located between the chain portion and the ligand portion described anywhere in this document.

[0540] In some implementation schemes: As shown in equation (I) anywhere in this document. It is any one of formula (VIa), (VIb) or (VIc), preferably formula (VIa): Formula (VIa) in: A I The protecting group is hydrogen or a suitable hydroxyl group; a is an integer, either 2 or 3; and b is an integer between 2 and 5; or Formula (VIb) in: A I The protecting group is hydrogen or a suitable hydroxyl group; a is an integer, either 2 or 3; c and d are independent integers from 1 to 6; or Formula (VIc) in: A I The protecting group is hydrogen or a suitable hydroxyl group; a is an integer, either 2 or 3; e is an integer between 2 and 10.

[0541] In some implementations, the portion shown in formula (I) For example (VIa): Formula (VIa) in: A I The protecting group is hydrogen or a suitable hydroxyl group; a is 3; and b is an integer 3.

[0542] In some implementations, as shown in formula (I) anywhere herein For equation (VII): Equation (VII) in: A I It is hydrogen; a is an integer, either 2 or 3, preferably 3.

[0543] Other exemplary compounds of this application include a “connector portion” as shown in formula (I*), which is part of the whole “connector”.

[0544] Formula I* in: r and s are independently selected from integers from 1 to 16; and Z is the oligonucleotide moiety.

[0545] As further understood in the art, exemplary compounds of this application comprise an entire connector located between the oligonucleotide moiety and the ligand moiety of these compounds. The entire connector “connects” the oligonucleotide moiety and the ligand moiety to each other.

[0546] The entire linker is generally assumed to contain one or more linker building blocks. For example, there exists a linker portion, described as a “linker portion” as represented by formula (I*), which is located adjacent to the ligand portion and typically connects the ligand portion directly or indirectly to the oligonucleotide portion via a branch point. The linker portion shown in formula (I*) is also often referred to as the “ligand arm” of the entire linker. Other linker portions may also exist between the oligonucleotide portion and the branch point (but not always), which are often referred to as the “tethering portion” of the entire linker, “tethering” the oligonucleotide portion to the remainder of the conjugated compound. Such 'ligand arms’ and / or 'linker portions’ and / or 'tethering portions’ can be understood with reference to the linear and / or branched configurations described above.

[0547] As can be seen from the claims and patent specification, the scope of this application covers linear or branched configurations, and there is no limitation on the number of individual ligands that may be present. Furthermore, those skilled in the art, based on prior art and the expertise of oligonucleotide chemists, will recognize that many structures can be used as linker sites.

[0548] The remaining portion of the entire connector (excluding the connector portion) listed in the claims, as well as the remainder of the patent specification, is represented by the chemical composition in formula (I), which the inventors consider to be particularly unique to this application. However, more generally, these chemical compositions can be described as the “tethered portion” as previously described, where the “tethered portion” is a part of the entire connector comprising the atomic group between Z (i.e., the oligonucleotide portion) and the connector portion, as shown in formula (I).

[0549] Chain section In the case of formula (I*), the “chain part” comprises the group of atoms between Z (i.e., the oligonucleotide part) and the linker part.

[0550] In some implementations, s is an integer selected from 4 to 12. In some implementations, s is 6.

[0551] In some implementations, r is an integer selected from 4 to 14. In some implementations, r is 6. In some implementations, r is 12.

[0552] In some implementations, r is 12 and s is 6.

[0553] Therefore, in some embodiments, the exemplary compounds of this application comprise the following structures: Equation (II*) In some implementations, r is 6 and s is 6.

[0554] Therefore, in some embodiments, the exemplary compounds of this application comprise the following structures: Equation (III*) Connector section With regard to formula (I*), the “connector portion” shown in formula (I) comprises the set of atoms located between the connector portion and the ligand portion described anywhere in this document.

[0555] In some implementations, such as the portion shown in formula (I*) described anywhere herein. It is any one of formula (IV*), (V*) or (VI*), preferably formula (IV*): Formula (IV*) in: A I The protecting group is hydrogen or a suitable hydroxyl group; a is an integer, either 2 or 3; and b is an integer between 2 and 5; or Formula (V*) in: A I The protecting group is hydrogen or a suitable hydroxyl group; a is an integer, either 2 or 3; and c and d are independent integers from 1 to 6; or Formula (VI*) Among them: A I It is a hydrogen or suitable hydroxyl protecting group; a is an integer of 2 or 3; and e is an integer from 1 to 10.

[0556] In some implementations, the portion shown in formula (I) For example (VIa*): Formula (VIa*) Among them: A I It is a hydrogen or a suitable hydroxyl protecting group; a is 3; and b is an integer 3.

[0557] In some implementations, the portion shown in formula (I) For equation (VII*): Equation (VII*) Among them: A I It represents hydrogen; a is an integer, 2 or 3.

[0558] In some implementations, a=2. In some implementations, a=3. In some implementations, b=3.

[0559] carriers and cells In one aspect, this application provides a cell containing nucleic acids (such as the repressive RNA [RNAi] described herein).

[0560] In one respect, this application provides a cell comprising the vector described herein.

[0561] In one aspect, this application provides a vector containing an oligonucleotide inhibitor such as iRNA (e.g., siRNA).

[0562] Pharmaceutically acceptable compositions In one aspect, this aspect provides pharmaceutical compositions for inhibiting the expression of target genes, said compositions comprising inhibitors such as oligomers, such as nucleic acids disclosed herein.

[0563] Pharmaceutically acceptable compositions may contain excipients and / or carriers.

[0564] Some examples of materials that can be used as pharmaceutically acceptable carriers include: (1) sugars, such as lactose, glucose and sucrose; (2) starches, such as corn starch and potato starch; (3) cellulose and its derivatives, such as sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate; (4) tragacanth gum powder; (5) malt; (6) gelatin; (7) lubricants, such as magnesium stearate, sodium lauryl sulfate and talc; (8) excipients, such as cocoa butter and suppository wax; (9) oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil; (10) glycols, such as propylene glycol; (11) (12) Polyols, such as glycerol, sorbitol, mannitol and polyethylene glycol; (13) Esters, such as ethyl oleate and ethyl laurate; (14) Agar; (15) Buffers, such as magnesium hydroxide and aluminum hydroxide; (16) Alginate; (17) Atherless water; (18) Isotonic saline; (19) Ringer's solution; (10) Ethanol; (21) pH buffer solution; (22) Polyesters, polycarbonates and / or polyanhydrides; (23) Compatibilizers, such as peptides and amino acids; (24) Serum components, such as serum albumin, HDL and LDL; and (25) Other non-toxic compatible substances used in pharmaceutical preparations.

[0565] Typical drug carriers include, but are not limited to, binders (e.g., pregelatinized corn starch, polyvinylpyrrolidone, or hydroxypropyl methylcellulose); fillers (e.g., lactose and other sugars, microcrystalline cellulose, pectin, gelatin, calcium sulfate, ethyl cellulose, polyacrylate, or dicalcium phosphate); lubricants (e.g., magnesium stearate, talc, silica, colloidal silica, stearic acid, metal stearate, hydrogenated vegetable oil, corn starch, polyethylene glycol, sodium benzoate, sodium acetate); disintegrants (e.g., starch, sodium carboxymethyl starch); and wetting agents (e.g., sodium dodecyl sulfate).

[0566] Pharmaceutically acceptable organic or inorganic excipients suitable for non-parenteral application and that do not react harmfully with nucleic acids may also be used to formulate the compositions of this application. Suitable pharmaceutically acceptable excipients include, but are not limited to, water, salt solutions, alcohols, polyethylene glycol, gelatin, lactose, amylose, magnesium stearate, talc, silicic acid, viscous paraffin, hydroxymethyl cellulose, polyvinylpyrrolidone, etc.

[0567] Formulations for topical application of nucleic acids may comprise sterile and non-sterile aqueous solutions, non-aqueous solutions in common solvents (e.g., alcohols), or nucleic acid solutions in liquid or solid oil bases. The solutions may also contain buffers, diluents, and other suitable additives. Pharmaceutically acceptable organic or inorganic excipients suitable for non-parenteral application and that do not cause adverse reactions with nucleic acids may also be used.

[0568] In one embodiment, the nucleic acid or composition is administered in a non-buffered solution. In some embodiments, the non-buffered solution is saline or water. In other embodiments, the nucleic acid (e.g., an RNAi reagent) is administered in a buffered solution. In such embodiments, the buffered solution may contain acetate, citrate, prolyl, carbonate, or phosphate, or any combination thereof. For example, the buffered solution may be a phosphate-buffered saline (PBS).

[0569] dose The pharmaceutical composition of this application can be administered at a dose sufficient to inhibit gene expression or alter the expression or function of a target (such as LNCRNA). Generally, when the composition contains nucleic acid, the appropriate nucleic acid (such as the siRNA of this application) dose ranges from about 0.001 to about 200.0 mg per kilogram of body weight per day for the subject, typically about 1-50 mg per kilogram of body weight per day. Typically, the appropriate nucleic acid (such as the siRNA of this application) dose ranges from about 0.1 mg / kg to about 5.0 mg / kg, for example, from about 0.3 mg / kg to about 3.0 mg / kg.

[0570] Repeated dosing regimens may involve periodic administration of therapeutic doses of nucleic acids, such as siRNA, for example, every other day or once a year. In some embodiments, nucleic acids, such as siRNA, are administered approximately once a month to approximately once a quarter (i.e., once every three months).

[0571] In various embodiments, the nucleic acid, such as siRNA, is administered at a dose of about 0.01 mg / kg to about 10 mg / kg or about 0.5 mg / kg to about 50 mg / kg. In some embodiments, the nucleic acid, such as siRNA, is administered at a dose of about 10 mg / kg to about 30 mg / kg. In some embodiments, the nucleic acid, such as siRNA, is administered at a dose selected from about 0.5 mg / kg, 1 mg / kg, 1.5 mg / kg, 3 mg / kg, 5 mg / kg, 10 mg / kg, and 30 mg / kg. In some embodiments, the nucleic acid, such as siRNA, is administered at a dose of about 0.1 mg / kg to about 5.0 mg / kg about once a week, once a month, once every two months, or once a quarter (i.e., once every three months). In some embodiments, the nucleic acid, such as siRNA, is administered to the subject once a week. In some embodiments, the nucleic acid, such as siRNA, is administered to the subject once a month. In some embodiments, the nucleic acid, such as siRNA, is administered once a quarter (i.e., once every three months).

[0572] After the initial treatment regimen, it can be administered at a less frequent frequency. For example, after three months of weekly or bi-weekly administration, it can be repeated monthly for six months or a year; or even longer.

[0573] The pharmaceutical composition can be administered once daily, or in two, three, or more fractions at appropriate intervals throughout the day, or even via continuous infusion or delivery by a controlled-release formulation. In this case, the amount of nucleic acid (e.g., siRNA) contained in each fraction needs to be reduced accordingly to achieve the total daily dose. Dosage units can also be formulated for continuous delivery over several days, for example using conventional sustained-release formulations to allow the sustained release of nucleic acids such as siRNA over several days. Sustained-release formulations are well known in the art and are particularly suitable for delivering pharmaceutical agents to a specific site, and can be used in the pharmaceutical agents described in this application. In this embodiment, the dosage unit comprises a corresponding multiple of the daily dose.

[0574] In other embodiments, a single dose of the pharmaceutical composition may have a long-lasting effect, such that subsequent doses are administered at intervals not exceeding 3, 4, or 5 days, or at intervals not exceeding 1, 2, 3, or 4 weeks. In some embodiments of this application, a single dose of the pharmaceutical composition is administered once weekly. In other embodiments of this application, a single dose of the pharmaceutical composition is administered once every two months. In some embodiments, the siRNA is administered approximately once monthly to once quarterly (i.e., approximately once every three months), or even once every six months or twelve months.

[0575] As is known in the art, the effective dose and in vivo half-life of the individual nucleic acids covered by this application, such as siRNA, can be estimated using conventional methods or based on in vivo experiments using suitable animal models.

[0576] The pharmaceutical composition of this application can be administered in a variety of ways, depending on whether local or systemic treatment is required, and depending on the area to be treated. It can be administered topically (e.g., via a transdermal patch); pulmonaryly, such as by inhalation or blowing in powder or aerosol, including via a nebulizer; intratracheal, intranasal, epidermal and transdermal, oral, or parenteral administration. Parenteral administration includes intravenous, intra-arterial, subcutaneous, intraperitoneal, or intramuscular injection or infusion; subcutaneous administration, such as via an implanted device; or intracranial administration, such as via intraparenchymal, intrathecal, or intraventricular administration. In some preferred embodiments, the composition is administered by intravenous infusion or injection. In some embodiments, the composition is administered by subcutaneous injection.

[0577] In one implementation, the nucleic acid, such as siRNA, is administered subcutaneously to the subject.

[0578] Inhibitors, such as nucleic acids like siRNA, can be delivered in a way that targets specific tissues (e.g., hepatocytes in particular).

[0579] Methods to inhibit gene expression or suppress target expression or function This application also provides methods for inhibiting gene expression in cells, as well as methods for inhibiting the expression and / or function of other target molecules such as LNCRNAs. The methods include contacting cells with an amount of the present application's nucleic acid, such as siRNA reagent (e.g., double-stranded siRNA), sufficient to inhibit gene expression in the cells, thereby inhibiting gene expression. In a preferred embodiment, the gene encodes an enzyme involved in post-translational glycosylation. In a more preferred embodiment, the gene is B4GALT1.

[0580] Contact between cells and inhibitors, such as nucleic acids (e.g., siRNA, like double-stranded siRNA reagents), can be performed in vitro or in vivo. In vivo contact of cells with inhibitory nucleic acids (e.g., siRNA) includes contacting cells or cell populations within a subject (e.g., a human subject) with the nucleic acid (e.g., siRNA). In vitro and in vivo cell contact methods can also be used in combination. As mentioned above, cell contact can be direct or indirect. Furthermore, cell contact can be achieved by targeting ligand portions, including any ligand portions described herein or known in the art. In a preferred embodiment, the targeting ligand portion is a carbohydrate portion (e.g., GalNAc3 ligand), or any other ligand portion capable of guiding the siRNA reagent to a target site.

[0581] The term “inhibition” as used in this article is used interchangeably with “reduction,” “silence,” “downregulation,” “curb,” and other similar terms, and includes any level of inhibition.

[0582] In some embodiments of the method described in this application, the expression or activity of a gene or repressive target (e.g., LNCRNA) is inhibited by at least 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 95%, or inhibited below the lower limit of the detection method, preferably measured by qPCR as described herein, and / or by transfecting siRNA into target cells. In some embodiments, the method includes clinically relevant inhibition of target gene expression, for example, as confirmed by observing clinically relevant results after treating a subject with an agent that reduces the expression of the gene and / or the activity of the target.

[0583] In some embodiments, when the nucleic acid of this application is transfected into cells, its IC50 value for inhibiting B4GALT1 gene expression is lower than 2500 pM, 2400 pM, 2300 pM, 2200 pM, 2100 pM, 2000 pM, 1900 pM, 1800 pM, 1700 pM, 1600 pM, 1500 pM, 1400 pM, 1300 pM, 1200 pM, 1100 pM, 1000 pM, 900 pM, 800 pM, 700 pM, 600 pM, 500 pM, 400 pM, 300 pM, 200 pM, or 100 pM, wherein the IC50 value is preferably determined by qPCR as described herein, and more preferably by reverse transcriptase (RT)-qPCR.

[0584] In a preferred embodiment, when the nucleic acid of this application is transfected into cells, its IC50 value for inhibiting B4GALT1 gene expression is less than 2500 pM. In a more preferred embodiment, when transfected into cells, the IC50 value of the nucleic acid of this application for inhibiting B4GALT1 gene expression is less than 1000 pM. In a more preferred embodiment, when transfected into cells, the IC50 value of the nucleic acid of this application for inhibiting B4GALT1 gene expression is less than 500 pM. In the most preferred embodiment, when transfected into cells, the IC50 value of the nucleic acid of this application for inhibiting B4GALT1 gene expression is less than 100 pM.

[0585] The inhibition of B4GALT1 gene expression can be quantitatively measured using the following methods: Huh7 cells (human hepatocyte-derived cell line, purchased from JCRB cell bank) were cultured in DMEM medium supplemented with 10% FBS at 37°C and 5% CO2 atmosphere. Cells were then transfected with a double-stranded siRNA targeting B4GALT1 mRNA or a negative control siRNA (siRNA-control; sense strand 5'-UUCUCCGAACGUGUCACGUTT-3' (SEQ ID NO:623), antisense strand 5'-ACGUGACACGUUCGGAGAATT-3' (SEQ ID NO:622)). Serial dilutions of 10×3 were used during transfection to achieve a final double-stranded concentration of 20 nM–1 pM. The transfection procedure involved adding 9.7 µL of Opti-MEM (ThermoFisher) and 0.3 µL of Lipofectamine RNAiMAX (ThermoFisher) to each 10 µL siRNA double-stranded sample. The mixture was incubated at room temperature for 15 minutes, then added to 100 µL of complete growth medium containing 20,000 Huh7 cells. Cells were incubated at 37°C / 5% CO2 for 24 hours, followed by purification of total RNA using the RNeasy 96 Kit (Qiagen). Each duplex was transfected in duplicate in a single experiment.

[0586] cDNA synthesis can be performed using the FastKing RT (containing gDNase) Kit (Tiangen). Real-time quantitative PCR (qPCR) can be performed on an ABI Prism 7900HT or ABI QuantStudio 7 using specific primers for human B4GALT1 (Hs00155245_m1) and human GAPDH (Hs02786624_g1) using the TaqMan Gene Expression Assay Kit (ThermoFisher Scientific).

[0587] qPCR can be repeated for cDNA from each well, and the average cycle threshold (Ct) can be calculated. The relative expression level of the B4GALT1 gene can be compared by Ct( The Ct method calculates the average Ct value, normalizes the expression using GAPDH as an internal reference gene, and uses untreated cells as a reference. The maximum inhibition rate and IC50 value of B4GALT1 expression can be calculated using a four-parameter (variable slope) model in GraphPad Prism 9 software.

[0588] As an alternative or supplementary method, the inhibitory potential of the nucleic acid in this application can be quantitatively determined without prior transfection of the nucleic acid into target cells.

[0589] Therefore, in some embodiments, when cells are co-incubated with the nucleic acid of this application, the EC50 value of the nucleic acid inhibiting B4GALT1 gene expression is lower than 1000 nM, 900 nM, 800 nM, 700 nM, 600 nM, 500 nM, 400 nM, 300 nM, 200 nM or 100 nM, wherein the EC50 value is preferably determined by qPCR as described herein, and more preferably by reverse transcriptase (RT)-qPCR as described herein.

[0590] In a preferred embodiment, when cells are co-incubated with the nucleic acid of this application, the EC50 value of the nucleic acid inhibiting B4GALT1 gene expression is less than 1000 nM. In a more preferred embodiment, when cells are co-incubated with the nucleic acid of this application, the EC50 value of the nucleic acid inhibiting B4GALT1 gene expression is less than 500 nM. In a more preferred embodiment, when cells are co-incubated with the nucleic acid of this application, the EC50 value of the nucleic acid inhibiting B4GALT1 gene expression is less than 200 nM. In the most preferred embodiment, when cells are co-incubated with the nucleic acid of this application, the EC50 value of the nucleic acid inhibiting B4GALT1 gene expression is less than 100 nM.

[0591] The inhibition of B4GALT1 gene expression in the presence of free nucleic acid can be quantitatively determined using the following methods: Primary C57BL / 6 mouse hepatocytes (PMHs) can be freshly isolated using a two-step collagenase liver perfusion method. Cells can be cultured in DMEM (Gibco-11995-092) medium supplemented with FBS, penicillin / streptomycin, HEPES, and L-glutamine. Cells should be cultured in a humidified incubator at 37°C and 5% CO2. Within 2 hours of isolation, PMH cells can be seeded into standard 96-well tissue culture plates at a density of 36,000 cells / well. Dose-response analysis of PMH can be performed by directly incubating cells with GalNAc-siRNA at final concentrations of 1000 nM, 500 nM, 250 nM, 125 nM, 62.5 nM, 31.3 nM, 15.6 nM, 7.8 nM, 3.9 nM, and 1.95 nM under physiologically free uptake conditions. In control wells, cells can be incubated without GalNAc-siRNA. After 48 hours of incubation, cells can be collected for RNA extraction. This can be used... RNeasy Total RNA was extracted using the kit according to the manufacturer's instructions (Qiagen, Shanghai, China). After reverse transcription, real-time quantitative PCR was performed using an ABI Prism 7900HT to detect RNA. B4GALT1 mRNA The relative abundance of the target gene was normalized using the housekeeping gene GAPDH. Relative quantification of the target gene expression level in each test sample can be performed by comparing Ct values ​​(ΔΔCt). This method measures the difference in Ct values ​​(ΔCt) between the target gene and the housekeeping gene. The formula is as follows: ΔCt = average Ct value of B4GALT1 – average Ct value of GAPDH, ΔΔCt = ΔCt (sample) – average ΔCt (untreated control group), relative expression level of target gene mRNA = 2. -ΔΔCt .

[0592] As an alternative or supplementary method, the inhibition of B4GALT1 gene expression can also be characterized by a decrease in the average relative expression level of the B4GALT1 gene.

[0593] In some embodiments, when cells are transfected with 0.1 nM of the nucleic acid of this application, the average relative expression level of B4GALT1 is less than 1, 0.9, 0.8, 0.7, 0.6, 0.5 or 0.4, said average relative expression level preferably determined by qPCR as described herein, more preferably by reverse transcriptase (RT)-qPCR as described herein.

[0594] In some embodiments, when cells are transfected with 5 nM of the nucleic acid of this application, the average relative expression level of B4GALT1 is less than 1, 0.9, 0.8, 0.7, 0.6, 0.5 or 0.4; as described herein, the average relative expression level is preferably determined by qPCR, more preferably by reverse transcriptase (RT)-qPCR.

[0595] The average relative expression level of the B4GALT1 gene can be quantitatively determined using the following methods: Huh7 cells (human hepatocyte-derived cell line, purchased from the JCRB cell bank) were cultured at 37°C and 5% CO2 in DMEM medium supplemented with 10% FBS. Cells were then transfected with a double-stranded siRNA targeting B4GALT1 mRNA or a negative control siRNA (siRNA-control; sense strand 5'-UUCUCCGAACGUGUCACGUTT-3' (SEQ ID NO:623), antisense strand 5'-ACGUGACACGUUCGGAGAATT-3' (SEQ ID NO:622)), with final double-stranded concentrations of 5 nM and 0.1 nM, respectively. Transfection was performed by adding 9.7 µL of Opti-MEM (ThermoFisher) and 0.3 µL of LipofectamineRNAiMAX (ThermoFisher) to each 10 µL siRNA double-stranded sample. The mixture was incubated at room temperature for 15 minutes, then added to 100 µL of complete growth medium containing 20,000 Huh7 cells. Cells were incubated at 37°C / 5% CO2 for 24 hours, and total RNA was purified using the RNeasy96 Kit (Qiagen). Each duplex was transfected twice independently, with two replicates per assay.

[0596] cDNA synthesis can be performed using the FastKing RT (containing gDNase) Kit (Tiangen). Real-time quantitative PCR (qPCR) can be performed on an ABI Prism 7900HT or ABI QuantStudio 7 using specific primers for human B4GALT1 (Hs00155245_m1) and human GAPDH (Hs02786624_g1) using the TaqMan Gene Expression Assay Kit (ThermoFisher Scientific).

[0597] qPCR can be repeated for cDNA from each well, and the average cycle threshold (Ct) can be calculated. The relative expression level of B4GALT1 can be compared by Ct ( The Ct method was used, with GAPDH as the internal reference gene and untreated cells as the control, and the Ct value was calculated based on the average Ct value.

[0598] The inhibition of gene expression can be confirmed by a reduction in the amount of mRNA in the target gene compared to a suitable control group. The inhibition of target function can be confirmed by a decrease in target activity compared to a suitable control group.

[0599] In other implementations, inhibition of gene or other target expression can be assessed by reducing parameters that are functionally related to gene expression, such as protein expression or signaling pathways.

[0600] Methods for treating or preventing diseases related to gene expression / target expression or function This application also provides a method for reducing or inhibiting gene expression in cells, or reducing the expression or function of a target gene, using nucleic acids (such as the siRNA of this application) or compositions containing nucleic acids (such as the siRNA of this application). The method includes contacting cells with nucleic acids (such as the dsiRNA of this application) and maintaining the cells for a sufficient time to allow for the degradation of the gene mRNA transcript, thereby inhibiting the expression of the gene in the cells. The reduction in target gene expression or function can be assessed by any method known in the art. In a preferred embodiment, the gene encodes an enzyme involved in post-translational glycosylation. In a more preferred embodiment, the gene is B4GALT1.

[0601] In the method of this application, cells can be contacted in vitro or in vivo, i.e., the cells can be in the body of the subject.

[0602] Cells suitable for treatment using the methods described in this application can be any cells expressing disease-related target genes or targets, such as those in vascular diseases like cardiovascular diseases.

[0603] The in vivo method of this application may include administering to a subject a composition comprising a nucleic acid (such as siRNA of this application), wherein the nucleic acid (such as siRNA) comprises a nucleoside sequence complementary to at least a portion of the RNA transcript of a gene of the mammal to be treated, or a sequence complementary to another nucleic acid whose expression and / or function is related to the disease.

[0604] This application also provides methods for treating subjects in need. The treatment methods described in this application include administering a therapeutically effective amount of the nucleic acid of this application, such as siRNA, to a subject; the subject being an individual capable of benefiting from reduced or inhibited expression of a specific gene, and / or reduced or inhibited expression and / or function of a target. The nucleic acid may be siRNA targeting a specific gene, or a pharmaceutical composition containing nucleic acid of the target gene. In one embodiment, the disease to be treated is a vascular disease, such as cardiovascular disease.

[0605] The nucleic acid (such as siRNA) of this application can be administered as "free" nucleic acid or "free siRNA," i.e., directly without the use of a pharmaceutical composition. The naked nucleic acid can be placed in a suitable buffer solution. The buffer solution may contain acetate, citrate, prolyl, carbonate, or phosphate, or any combination thereof. In one embodiment, the buffer solution is phosphate-buffered saline (PBS). The pH and osmotic pressure of the buffer solution can be adjusted to suit its administration to the subject.

[0606] As an alternative, the nucleic acid (e.g., siRNA) of this application can be administered as a pharmaceutical composition (e.g., dsiRNA liposome formulation).

[0607] In one embodiment, the method includes administering the composition described herein to reduce the expression of a target gene, for example, for a duration of about 1, 2, 3, 4, 5, 6, 7, 8, 12, 16, 18, 24, 28, 32, or about 36 hours. In one embodiment, the reduction in target gene expression lasts for a longer period, for example, at least about two, three, four days, or longer, for example, about one, two, three, or four weeks, or longer, for example, about one, two, or three months.

[0608] Therapeutic doses of nucleic acids, such as siRNA, can be administered to the subject, for example, from about 0.01 mg / kg to about 200 mg / kg.

[0609] Nucleic acids, such as siRNA, can be administered periodically via intravenous infusion over a period of time. In some embodiments, treatment can be administered at a lower frequency after an initial treatment regimen. Administration of siRNA can reduce, for example, the level of the gene product of a target gene in patient cells or tissues by at least about 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 95%, or below the detection limit of the detection method used. In some embodiments, the administration can lead to clinical stability of at least one sign or symptom of the gene-related disease, or preferably a clinically significant reduction.

[0610] As an alternative, nucleic acids (e.g., siRNA) can be administered subcutaneously, i.e., by subcutaneous injection. The required daily dose of nucleic acid, such as iRNA, can be delivered to the subject using one or more injections. Injections can be repeated over a period of time. Administration can be repeated periodically. In some embodiments, the frequency of treatment can be reduced after the initial treatment regimen. Repeated dosing regimens may include periodic administration of therapeutic doses of nucleic acid, such as every other day or once a year. In some embodiments, nucleic acid is administered approximately once a month to approximately once a quarter (i.e., approximately once every three months).

[0611] In one respect, this application is applicable to the compounds, methods, compositions, or uses described in statements 1-101 below (wherein, any formula mentioned in statements 1-101 refers only to the formulas defined in statements 1-101). These formulas are... Figure 6 (Recurrence in China).

[0612] 1. A compound having the following structure: Formula (I) in: R1 is selected independently from hydrogen, methyl, and ethyl each time it appears; R2 is selected from the following group: hydrogen, hydroxyl group, -OC 1-3 Alkyl group, -C(=O)OC 1-3 Alkyl, halogen, and nitro groups; X1 and X2 are each independently selected from methylene, oxygen, and sulfur when they appear; m is an integer from 1 to 6; n is an integer from 1 to 10; q, r, s, t, v are independent integers from 0 to 4, and satisfy the following condition: (i) q and r cannot both be 0; and (ii) s, t and v cannot all be 0 at the same time; Z is the oligonucleotide moiety.

[0613] 2. According to the compound described in statement 1, R1 is hydrogen each time it appears.

[0614] 3. The compound according to statement 1, wherein R1 is a methyl group.

[0615] 4. The compound according to statement 1, wherein R1 is an ethyl group.

[0616] 5. The compound according to any one of statements 1-4, wherein R2 is a hydroxyl group.

[0617] 6. The compound according to any one of statements 1-4, wherein R2 is a halogen.

[0618] 7. The compound according to statement 6, wherein R2 is fluorine.

[0619] 8. The compound according to statement 6, wherein R2 is chlorine.

[0620] 9. The compound according to statement 6, wherein R2 is bromine.

[0621] 10. The compound according to statement 6, wherein R2 is iodine.

[0622] 11. The compound according to statement 6, wherein R2 is a nitro group.

[0623] 12. The compound according to any one of statements 1-11, wherein X1 is a methylene group.

[0624] 13. The compound according to any one of statements 1-11, wherein X1 is oxygen.

[0625] 14. The compound according to any one of statements 1-11, wherein X1 is sulfur.

[0626] 15. The compound according to any one of statements 1-14, wherein X2 is a methylene group.

[0627] 16. The compound according to any one of statements 1-15, wherein X2 is oxygen.

[0628] 17. The compound according to any one of statements 1-16, wherein X2 is sulfur.

[0629] 18. The compound according to any one of clauses 1-17, wherein m=3.

[0630] 19. The compound according to any one of statements 1-18, wherein n=6.

[0631] 20. The compound according to statement 13 or 15, wherein X1 is oxygen and X2 is methylene, and preferably wherein: q=1, r=2, s=1, t=1, v=1.

[0632] 21. The compound according to statement 12 or 15, wherein X1 and X2 are both methylene groups, and preferably wherein: q=1, r=3, s=1, t=1, v=1.

[0633] 22. A compound according to any one of statements 1-21, wherein Z is: ; in: Z1, Z2, Z3, and Z4 each appear independently as either oxygen or sulfur; and The bonds between P and Z2, and between P and Z3, are a single bond and a double bond, respectively.

[0634] 23. The compound according to statement 22, wherein the oligonucleotide is an RNA compound capable of regulating (preferably inhibiting) the expression of a target gene.

[0635] 24. The compound according to statement 23, wherein the RNA compound comprises an RNA duplex containing a first strand and a second strand, wherein the first strand is at least partially complementary to the RNA sequence of the target gene, the second strand is at least partially complementary to the first strand, and wherein the first strand and the second strand each have a 5' end and a 3' end.

[0636] 25. The compound according to statement 24, wherein the RNA compound is attached to an adjacent phosphate group at the 5' end of its second strand.

[0637] 26. The compound according to statement 24, wherein the RNA compound is attached to an adjacent phosphate group at the 3' end of its second strand.

[0638] 27. Compound of formula (II): Equation (II).

[0639] 28. Compound of formula (III): Formula (III).

[0640] 29. The compound according to statement 27 or 28, wherein the oligonucleotide comprises an RNA duplex containing a first strand and a second strand, wherein the first strand is at least partially complementary to the RNA sequence of the target gene, the second strand is at least partially complementary to the first strand, and the first strand and the second strand each have a 5' end and a 3' end, the RNA duplex being linked to an adjacent phosphate group at the 5' end of its second strand.

[0641] 30. A composition comprising a compound of formula (II) as defined in statement 27 and a compound of formula (III) as defined in statement 28, optionally comprising a compound as described in statement 29.

[0642] 31. The composition according to statement 30, wherein the content of the compound of formula (III) as defined in statement 28 is 10%-15% of the weight of the composition.

[0643] 32. Compounds of formula (IV): Formula (IV).

[0644] Compound of formula (V): Equation (V).

[0645] 34. The compound according to statement 32 or 33, wherein the oligonucleotide comprises an RNA duplex containing a first strand and a second strand, wherein the first strand is at least partially complementary to the RNA sequence of the target gene, the second strand is at least partially complementary to the first strand, wherein the first strand and the second strand each have a 5' end and a 3' end, and the RNA duplex is linked to an adjacent phosphate group at the 3' end of its second strand.

[0646] 35. A composition comprising a compound of formula (IV) as defined in statement 32 and a compound of formula (V) as defined in statement 33; optionally, the composition comprises a compound as described in statement 34.

[0647] 36. The composition according to statement 35, wherein the content of the compound of formula (V) as defined in statement 33 is 10%-15% of the weight of the composition.

[0648] 37. A compound as defined in any one of statements 1-29 or 32-34, wherein the oligonucleotide comprises an RNA duplex, the RNA duplex further comprising one or more riboses modified at the 2' position, preferably comprising multiple riboses modified at the 2' position.

[0649] 38. The compound according to statement 37, wherein the modification is selected from 2'-O-methyl, 2'-deoxy-fluorine, and 2'-deoxy.

[0650] 39. The compound according to any one of statements 1-29, 32-34, or 37-38, wherein the oligonucleotide further comprises one or more degradation-protecting moieties at one or more ends.

[0651] 40. The compound according to statement 39, wherein the one or more degradation protecting moieties are not present at the end of the oligonucleotide chain carrying the ligand moieties, and / or wherein the one or more degradation protecting moieties are selected from thiophosphate nucleoside inter-linked bonds, dithiophosphate nucleoside inter-linked bonds, and reverse abasic nucleosides, wherein the reverse abasic nucleosides are located at the distal end of the chain carrying the ligand moieties.

[0652] 41. The compound according to any one of statements 1-29 or 32-34 or 37-40, wherein the ligand portion represented by formula (I) in statement 1 comprises one or more ligands.

[0653] 42. The compound according to statement 41, wherein the ligand portion represented by formula (I) in statement 1 comprises one or more carbohydrate ligands.

[0654] 43. The compound according to statement 42, wherein the one or more carbohydrates may be monosaccharides, disaccharides, trisaccharides, tetrasaccharides, oligosaccharides, or polysaccharides.

[0655] 44. The compound according to statement 43, wherein the one or more carbohydrates comprise one or more galactose moieties, one or more lactose moieties, one or more N-acetylgalactosamine moieties, and / or one or more mannose moieties.

[0656] 45. The compound according to statement 44, wherein the one or more carbohydrates comprise one or more N-acetylgalactosamine moieties.

[0657] 46. ​​The compound according to statement 45, comprising two or three N-acetylgalactosamine moieties.

[0658] 47. The compound according to any one of statements 41-46, wherein the one or more ligands are connected in a linear or branched configuration.

[0659] 48. The compound according to statement 47, wherein the one or more ligands are connected in a biantennary or triantennary branching configuration.

[0660] 49. The compound according to statements 46-48, wherein the portion as shown in formula (I) in statement 1: It can be any one of formula (VIa), formula (VIb) or formula (VIc), preferably formula (VIa): Formula (VIa); in: A I It is hydrogen, or a suitable hydroxyl protecting group; a is an integer, either 2 or 3; and b is an integer between 2 and 5; or Formula (VIb); in: A I It is hydrogen, or a suitable hydroxyl protecting group; a is an integer, either 2 or 3; and c and d are independent integers from 1 to 6; or Formula (VIc); Among them: A I The protecting group is hydrogen or a suitable hydroxyl group; a is an integer of 2 or 3; e is an integer from 2 to 10.

[0661] 50. The compound according to statements 46-48, wherein the portion as shown in formula (I) in statement 1: For equation (VII): Equation (VII); Among them: A I It is hydrogen; a is an integer, 2 or 3.

[0662] 51. The compound according to statement 49 or 50, wherein a = 2.

[0663] 52. The compound described in statement 49 or 50, wherein a = 3.

[0664] 53. According to the compound described in statement 49, where b=3.

[0665] 54. Compound of formula (VIII): Formula (VIII).

[0666] Compound of formula (IX): Formula (IX).

[0667] 56. The compound according to statement 54 or 55, wherein the oligonucleotide comprises an RNA duplex containing a first strand and a second strand, wherein the first strand is at least partially complementary to the RNA sequence of the target gene, the second strand is at least partially complementary to the first strand, wherein the first strand and the second strand each have a 5' end and a 3' end, and the RNA duplex is linked to an adjacent phosphate group at the 5' end of its second strand.

[0668] 57. A composition comprising a compound of formula (VIII) as defined in statement 54 and a compound of formula (IX) as defined in statement 55; optionally, the composition comprises the compound of statement 56.

[0669] 58. The composition according to statement 57, wherein the content of the compound of formula (IX) as defined in statement 55 is 10%-15% of the weight of the composition.

[0670] 59. Compound of formula (X): Formula (X).

[0671] Compound of formula (XI): Formula (XI).

[0672] 61. The compound according to statement 59 or 60, wherein the oligonucleotide comprises an RNA duplex containing a first strand and a second strand, wherein the first strand is at least partially complementary to the RNA sequence of the target gene, the second strand is at least partially complementary to the first strand, wherein the first strand and the second strand each have a 5' end and a 3' end, and the RNA duplex is linked to an adjacent phosphate group at the 3' end of its second strand.

[0673] 62. A composition comprising a compound of formula (X) as defined in statement 59 and a compound of formula (XI) as defined in statement 60; optionally, the composition comprises the compound of statement 61.

[0674] 63. The composition according to statement 62, wherein the content of the compound of formula (XI) as defined in statement 60 is 10%-15% of the weight of the composition.

[0675] 64. The compound as defined in any one of statements 54-63, wherein the oligonucleotide comprises an RNA duplex, the RNA duplex further comprising one or more riboses modified at the 2' position, preferably multiple riboses modified at the 2' position.

[0676] 65. The compound according to statement 64, wherein the modification is selected from 2'-O-methyl, 2'-deoxy-fluorine, and 2'-deoxy.

[0677] 66. The compound according to any one of statements 54-65, wherein the oligonucleotide further comprises one or more degradation-protecting moieties at one or more ends.

[0678] 67. The compound according to statement 66, wherein the one or more degradation protecting moieties are not present at the end of the oligonucleotide chain carrying the ligand moieties, and / or wherein the one or more degradation protecting moieties are selected from thiophosphate nucleoside inter-linked bonds, dithiophosphate nucleoside inter-linked bonds, and reverse abasic nucleosides, wherein the reverse abasic nucleosides are present at the distal end of the chain carrying the ligand moieties, as shown in any one of formula (VIII), (IX), (X), or (XI) of any one of statements 54, 55, 59, or 60.

[0679] 68. A method for preparing a compound according to any one of statements 1-29, 32-34, 37-56, 59-61 and 64-67, and / or a composition according to any one of statements 30, 31, 35, 36, 57, 58, 62, 63, said method comprising reacting a compound of formula (XII) and (XIII): Formula (XII) Formula (XIII); in: R1 is selected independently from hydrogen, methyl, and ethyl each time it appears; R2 is selected from the following group: hydrogen, hydroxyl group, -OC 1-3 Alkyl group, -C(=O)OC 1-3 Alkyl, halogen, and nitro groups; X1 and X2 are each independently selected from methylene, oxygen, and sulfur when they appear; m is an integer from 1 to 6; n is an integer from 1 to 10; q, r, s, t, v are independent integers from 0 to 4, provided that: (i) q and r cannot both be 0; and (ii) s, t and v cannot all be 0 at the same time; Z represents the oligonucleotide moiety; And, where appropriate, perform ligand deprotection and / or annealing of the second chain of the oligonucleotide moiety.

[0680] 69. The method according to statement 68, wherein compound (XII) is prepared by reacting compounds of formula (XIV) and formula (XV): Formula (XIV) Formula (XV); R1 is selected independently from hydrogen, methyl, and ethyl each time it appears; R2 is selected from the following group: hydrogen, hydroxyl group, -OC 1-3 Alkyl group, -C(=O)OC 1-3 Alkyl, halogen, and nitro groups; X1 and X2 are each independently selected from methylene, oxygen, and sulfur when they appear; q, r, s, t, v are independent integers from 0 to 4, provided that: (i) q and r cannot both be 0; and (ii) s, t and v cannot all be 0 at the same time; Z represents the oligonucleotide moiety.

[0681] 70. The method according to statement 68 is used to prepare a compound according to any one of statements 20, 25, 27, 29, 54, and 56, and / or a composition according to any one of statements 30, 31, 57, and 58, wherein: Compound of formula (XII) is of formula (XIIa): Formula (XIIa) Compound (XIII) is of formula (XIIIa): Formula (XIIIa); The oligonucleotide includes an RNA duplex containing a first strand and a second strand, wherein the first strand is at least partially complementary to the RNA sequence of the target gene, the second strand is at least partially complementary to the first strand, both the first strand and the second strand have a 5' end and a 3' end, and the RNA duplex is connected to an adjacent phosphate group at the 5' end of its second strand.

[0682] 71. The method according to statement 68 is used to prepare a compound according to any one of statements 20, 25, 28, 29, 55, and 56, and / or a composition according to any one of statements 30, 31, 57, and 58, wherein: Compound (XII) is of formula (XIIb): Formula (XIIb); Furthermore, the compound of formula (XIII) is of formula (XIIIa): (XIIIa); The oligonucleotide comprises an RNA duplex containing a first strand and a second strand, wherein the first strand is at least partially complementary to the RNA sequence of the target gene, the second strand is at least partially complementary to the first strand, wherein the first strand and the second strand each have a 5' end and a 3' end, and the RNA duplex is connected to an adjacent phosphate group at the 5' end of its second strand.

[0683] 72. The method according to statement 68 is used to prepare a compound according to any one of statements 21, 26, 32, 34, 59, and 61, and / or a composition according to any one of statements 35, 36, 62, and 63, wherein: Compound of formula (XII) is of formula (XIIc): Formula (XIIc); Furthermore, the compound of formula (XIII) is of formula (XIIIa): Formula (XIIIa); The oligonucleotide comprises an RNA duplex containing a first strand and a second strand, wherein the first strand is at least partially complementary to the RNA sequence of the target gene, the second strand is at least partially complementary to the first strand, wherein both the first and second strands have a 5' end and a 3' end, and wherein the RNA duplex is connected to an adjacent phosphate group at the 3' end of its second strand.

[0684] 73. The method according to statement 68 is used to prepare a compound according to any one of statements 21, 26, 33, 34, 60, and 61, and / or a composition according to any one of statements 35, 36, 62, and 63, wherein: Compound of formula (XII) is of formula (XIId): Formula (XIId) And the compound of formula (XIII) is of formula (XIIIa): Formula (XIIIa); The oligonucleotide comprises an RNA duplex containing a first strand and a second strand, wherein the first strand is at least partially complementary to the RNA sequence of the target gene, the second strand is at least partially complementary to the first strand, wherein both the first strand and the second strand have a 5' end and a 3' end, and wherein the RNA duplex is connected to an adjacent phosphate group at the 3' end of its second strand.

[0685] 74. The method according to any one of statements 70-73, wherein: Compound (XIIIa) is of formula (XIIIb): Equation (XIIIb).

[0686] 75. The method described in statement 69 is subordinate to statements 70-73, wherein: Compounds of formula (XIV) are of formula (XIVa) or formula (XIVb): Formula (XIVa) Formula (XIVb); Furthermore, compounds of formula (XV) are of formula (XVa) or formula (XIVb): Equation (XVa) Formula (XVb); The oligonucleotide comprises an RNA duplex containing a first strand and a second strand, wherein the first strand is at least partially complementary to the RNA sequence of the target gene, and the second strand is at least partially complementary to the first strand, wherein both the first strand and the second strand have a 5' end and a 3' end, and wherein (i) the RNA duplex is linked at the 5' end of its second strand to an adjacent phosphate group of formula (XVa), or (ii) the RNA duplex is linked at the 3' end of its second strand to an adjacent phosphate group of formula (XVb).

[0687] 76. Compound of formula (XII): Formula (XII); in: R1 is selected independently from hydrogen, methyl, and ethyl each time it appears; R2 is selected from the following group: hydrogen, hydroxyl group, -OC 1-3 Alkyl group, -C(=O)OC 1-3 Alkyl, halogen, and nitro groups; X1 and X2 are each independently selected from methylene, oxygen, and sulfur when they appear; q, r, s, t, v are independent integers from 0 to 4, provided that: (i) q and r cannot both be 0; and (ii) s, t and v cannot all be 0 at the same time; Z is the oligonucleotide moiety.

[0688] 77. Compound of formula (XIIa): Formula (XIIa).

[0689] 78. Compound of formula (XIIb): Formula (XIIb).

[0690] 79. Compound of formula (XIIc): Formula (XIIc).

[0691] 80. Compound of formula (XIId): Formula (XIId).

[0692] 81. Compound of formula (XIII): Formula (XIII); in: R1 is independently selected from hydrogen, methyl, and ethyl in each occurrence; m is an integer from 1 to 6; n is an integer from 1 to 10.

[0693] 82. Compound of formula (XIIIa): Equation (XIIIa).

[0694] 83. Compound of formula (XIIIb): Equation (XIIIb).

[0695] 84. Compound of formula (XIV): Formula (XIV); in: R1 is selected from hydrogen, methyl, and ethyl; R2 is selected from the following group: hydrogen, hydroxyl group, -OC 1-3 Alkyl group, -C(=O)OC 1-3 Alkyl, halogen, and nitro groups; X2 is selected from methylene, oxygen, and sulfur; s, t, and v are integers from 0 to 4, but s, t, and v cannot all be 0 at the same time.

[0696] Compound of formula (XIVa): Formula (XIVa).

[0697] 86. Compound of formula (XIVb): Formula (XIVb).

[0698] Compound of formula (XV): Formula (XV); in: R1 is selected independently from hydrogen, methyl, and ethyl each time it appears; X1 is selected from methylene, oxygen, and sulfur; q and r are independent integers from 0 to 4, provided that q and r cannot both be 0 at the same time; Z is the oligonucleotide moiety.

[0699] Compound of formula (XVa): Formula (XVa).

[0700] Compound of formula (XVb): Formula (XVb).

[0701] 90. Use of the compound according to any one of statements 76, 81-84, 87 in the preparation of the compound according to any one of statements 1-29, 32-34, 37-56, 59-61 and 64-67, and / or the composition according to any one of statements 30, 31, 35, 36, 57, 58, 62 and 63.

[0702] 91. Use of the compound according to statement 85 in the preparation of the compound of any one of statements 1-29, 32-34, 37-56, 59-61 and 64-67, and / or the composition of any one of statements 30, 31, 35, 36, 57, 58, 62 and 63, wherein R2=F.

[0703] 92. Use of the compound according to statement 86 in the preparation of the compound of any one of statements 1-29, 32-34, 37-56, 59-61 and 64-67, and / or the composition of any one of statements 30, 31, 35, 36, 57, 58, 62 and 63, wherein R2=OH.

[0704] 93. Use of the compound according to statement 77 in the preparation of the compound of any one of statements 20, 25, 27, 29, 54, 56, and / or the composition of any one of statements 30, 31, 57, 58.

[0705] 94. Use of the compound according to statement 78 in the preparation of the compound of any one of statements 20, 25, 28, 29, 55, 56, and / or the composition of any one of statements 30, 31, 57, 58.

[0706] 95. Use of the compound according to statement 79 in the preparation of the compound of any one of statements 21, 26, 32, 34, 59, 61, and / or the composition of any one of statements 35, 36, 62, 63.

[0707] 96. Use of the compound according to statement 80 in the preparation of the compound of any one of statements 21, 26, 33, 34, 60, 61, and / or any one of the compositions of statements 35, 36, 62, 63.

[0708] 97. Use of the compound according to statement 88 in the preparation of the compound of any one of statements 20, 25, 27-29, 54-56, and / or the composition of any one of statements 30, 31, 57, 58.

[0709] 98. Use of the compound according to statement 89 in the preparation of the compound of any one of statements 21, 26, 32-34, 59-61, and / or the composition of any one of statements 35, 36, 62, 63.

[0710] 99. A compound or composition obtained or available by any one of statements 68-75.

[0711] 100. A pharmaceutical...

Claims

1. Use of inhibitors of B4GALT1 expression and / or function in the prevention and / or treatment and / or control of insulin resistance, and / or elevated blood glucose levels, and / or elevated blood insulin levels, and / or elevated blood HbA1c levels, and / or elevated blood free fatty acid levels, and / or elevated blood fibrinogen levels, and / or elevated blood total cholesterol levels, and / or elevated blood LDL cholesterol levels, and / or elevated blood triglyceride levels.

2. Use of inhibitors of B4GALT1 expression and / or function in the prevention and / or treatment and / or control of insulin resistance, and / or elevated blood glucose levels, and / or elevated blood insulin levels, and / or elevated blood HbA1c levels, and / or elevated blood free fatty acid levels, and / or elevated blood fibrinogen levels, and / or elevated blood total cholesterol levels, and / or elevated blood LDL cholesterol levels, and / or elevated blood triglyceride levels, and / or diabetes in patients with or at risk of developing metabolic and / or vascular diseases.

3. The use of the inhibitor according to claim 1 or 2 in the prevention and / or treatment and / or control of insulin resistance; preferably, wherein the inhibitor is capable of improving insulin resistance.

4. Use of the inhibitor according to any one of claims 1-3 in the prevention and / or treatment and / or control of elevated blood glucose levels; preferably, the inhibitor is capable of reducing elevated blood glucose levels.

5. Use of the inhibitor according to any one of claims 1-4 in the prevention and / or treatment and / or control of elevated blood insulin levels; preferably, said inhibitor is capable of reducing elevated blood insulin levels.

6. Use of the inhibitor according to any one of claims 1-5 in the prevention and / or treatment and / or control of elevated serum HbA1c levels; preferably, said inhibitor is capable of reducing elevated serum HbA1c levels.

7. Use of the inhibitor according to any one of claims 1-6 in the prevention and / or treatment and / or control of elevated blood free fatty acid levels; preferably, said inhibitor is capable of reducing elevated blood free fatty acid levels.

8. Use of the inhibitor according to any one of claims 1-7 in the prevention and / or treatment and / or control of elevated fibrinogen levels; preferably, said inhibitor is capable of reducing elevated fibrinogen levels.

9. Use of the inhibitor according to any one of claims 1-8 in the prevention and / or treatment and / or control of elevated total cholesterol levels; preferably, wherein the inhibitor is capable of reducing elevated total cholesterol levels.

10. Use of the inhibitor according to any one of claims 1-9 in the prevention and / or treatment and / or control of elevated blood LDL cholesterol levels; preferably, said inhibitor is capable of reducing elevated blood LDL cholesterol levels.

11. Use of the inhibitor according to any one of claims 1-10 in the prevention and / or treatment and / or control of elevated blood triglyceride levels; preferably, said inhibitor is capable of reducing elevated blood triglyceride levels.

12. Use of inhibitors of B4GALT1 expression and / or function in the prevention and / or treatment and / or control of vascular disease in patients, wherein, The vascular disease is associated with insulin resistance, and / or elevated levels of free fatty acids, and / or elevated levels of fibrinogen, and / or elevated levels of total cholesterol, and / or elevated levels of LDL cholesterol, and / or elevated levels of triglycerides, and / or diabetes.

13. Use of the inhibitor according to claim 12 in the prevention and / or treatment and / or control of vascular diseases associated with insulin resistance; preferably, the inhibitor is capable of improving insulin resistance.

14. Use of the inhibitor according to claim 12 or 13 in the prevention and / or treatment and / or control of vascular diseases associated with elevated blood free fatty acid levels; preferably, wherein the inhibitor is capable of reducing elevated blood free fatty acid levels.

15. Use of the inhibitor according to any one of claims 12-14 in the prevention and / or treatment and / or control of vascular disease associated with elevated fibrinogen levels; preferably, wherein the inhibitor is capable of reducing elevated fibrinogen levels.

16. Use of the inhibitor according to any one of claims 12-15 in the prevention and / or treatment and / or control of vascular disease associated with elevated total cholesterol levels; preferably, wherein the inhibitor is capable of reducing elevated total cholesterol levels.

17. Use of the inhibitor according to any one of claims 12-16 in the prevention and / or treatment and / or control of vascular disease associated with elevated blood LDL cholesterol levels; preferably, wherein the inhibitor is capable of reducing elevated blood LDL cholesterol levels.

18. Use of the inhibitor according to any one of claims 12-17 in the prevention and / or treatment and / or control of vascular disease associated with elevated blood triglyceride levels; preferably, said inhibitor is capable of reducing elevated blood triglyceride levels.

19. Use of the inhibitor according to any one of claims 12-18 in the prevention and / or treatment and / or control of vascular disease associated with diabetes; preferably, wherein the inhibitor is capable of improving diabetes.

20. Use of inhibitors of B4GALT1 expression and / or function in the prevention and / or treatment and / or control of obesity, and / or weight gain, and / or metabolic syndrome in patients.

21. The inhibitor for the use according to claim 20, wherein, The obesity, and / or weight gain, and / or metabolic syndrome are associated with insulin resistance, and / or elevated blood free fatty acid levels; preferably, the inhibitor is capable of reducing elevated blood free fatty acid levels.

22. The inhibitor for the use according to claim 21, wherein, The obesity, and / or weight gain, and / or metabolic syndrome are associated with insulin resistance.

23. The inhibitor for the use according to claim 20 or 21, wherein, The obesity, and / or weight gain, and / or metabolic syndrome are associated with elevated blood free fatty acid levels; preferably, the inhibitor is capable of reducing elevated blood free fatty acid levels.

24. The inhibitor for the use according to any one of claims 20-23, wherein, The obesity, and / or weight gain, and / or metabolic syndrome are associated with elevated total cholesterol levels, and / or elevated LDL cholesterol levels, and / or elevated triglyceride levels; preferably, the inhibitor is capable of reducing elevated total cholesterol levels, and / or elevated LDL cholesterol levels, and / or elevated triglyceride levels.

25. The inhibitor for the use according to any one of claims 20-24, wherein, The metabolic syndrome is further or independently associated with elevated total cholesterol levels, and / or elevated LDL cholesterol levels, and / or elevated triglyceride levels; preferably, the inhibitor is capable of reducing elevated total cholesterol levels, and / or elevated LDL cholesterol levels, and / or elevated triglyceride levels.

26. The inhibitor for the use according to any one of claims 20-25, wherein, The metabolic syndrome is further or independently associated with elevated total cholesterol levels; preferably, the inhibitor is capable of reducing elevated total cholesterol levels.

27. The inhibitor for the use according to any one of claims 20-26, wherein, The metabolic syndrome is further or independently associated with elevated blood LDL cholesterol levels; preferably, the inhibitor is capable of reducing elevated blood LDL cholesterol levels.

28. The inhibitor for the use according to any one of claims 20-27, wherein, The metabolic syndrome is further or independently associated with elevated blood triglyceride levels; preferably, the inhibitor is capable of reducing elevated blood triglyceride levels.

29. An inhibitor for the use according to any one of the preceding claims, wherein, The patient is a mammalian patient, preferably a human patient.

30. An inhibitor for the use according to any one of the preceding claims, wherein, The inhibitor suppresses the expression of B4GALT1 in hepatocytes.

31. An inhibitor for the use according to any one of the preceding claims, wherein, The inhibitor of B4GALT1 expression and / or function is an siRNA oligomer.

32. The inhibitor for the use according to claim 30 or 31, wherein, The siRNA oligomer is conjugated to one or more ligands.

33. The inhibitor for the use according to claim 32, wherein, The one or more ligand portions comprise one or more GalNAc ligands and / or one or more GalNAc ligand derivatives.

34. An inhibitor for the use described in any one or more of the preceding claims, wherein it is an siRNA oligonucleotide having a first strand and a second strand, wherein: i) The first strand of the siRNA has a length of 15-30 nucleotides, preferably 19-25 nucleotides, more preferably 23 or 25 nucleotides; even more preferably 23 nucleotides; and / or ii) The second strand of siRNA has a length of 15-30 nucleotides, preferably 19-25 nucleotides, and more preferably 21 nucleotides.

35. The inhibitor for the use according to claim 34, wherein, The second positive chain further includes one or more abasic nucleosides in its terminal region, wherein the abasic nucleosides are connected to adjacent nucleosides via reverse internucleotide bonds.

36. The inhibitor for the use according to claim 35, wherein, The second chain includes: i. Two or more abase-free nucleotides located in the terminal region of the second chain; and / or ii. Two or more abase-free nucleosides located in the 5' or 3' end region of the second strand; and / or iii. Two or more abasic nucleosides located in the 5' or 3' end region of the second chain, wherein the abasic nucleosides are located at the overhangs described herein; and / or iv. Two or more consecutive abasic nucleosides are present in the terminal region of the second chain; preferably, one of the abasic nucleosides is a terminal nucleoside; and / or v. Two or more consecutive abasic nucleosides are present in the 5' or 3' end region of the second strand; preferably, one of the abasic nucleosides is a terminal nucleoside in the 5' or 3' end region of the second strand; and / or vi. In the terminal region of the second chain, at least one base-free nucleotide is linked to an adjacent base-containing nucleotide via a reverse internucleotide bond; and / or vii. In the 5' or 3' terminal region of the second strand, at least one base-free nucleotide is linked to an adjacent base-containing nucleotide via a reverse internucleotide bond; and / or viii. The penultimate nucleoside is a baseless nucleoside, which is linked to a non-terminal nucleoside (referred to as the penultimate nucleoside in this paper) via a reverse bond; and / or ix. When read along the chain towards the end, the two terminal nucleosides are baseless nucleosides and are linked by a 5'-3' bond; x. When read along the chain towards the end containing the terminal nucleosides, the two terminal nucleosides are baseless nucleosides and are linked by 3'-5' bonds; xi. The last two positions are nucleosides without bases, wherein the penultimate nucleoside and the penultimate nucleoside are connected by a reverse bond, and the reverse bond is a 5'-5' reverse bond or a 3'-3' reverse bond; xii. The last two positions are occupied by nucleosides without bases, wherein the penultimate nucleoside and the penultimate nucleoside are linked by an anti-bond, and wherein... (1) The reverse bond is a 5'-5' reverse bond, and when read along the direction including the end and the penultimate abasic nucleotide, the connection between the end and the penultimate abasic nucleotide is a 3'-5' bond; or (2) The reverse bond is a 3'-3' reverse bond, and when read along the end direction containing the end and the penultimate abase nucleoside, the connection between the end and the penultimate abase nucleoside is a 5'-3' bond.

37. The inhibitor for the use according to claim 35 or 36, wherein, The reverse nucleoside inter-bond is located in a terminal region far from the 5' end region of the second strand, or in a terminal region far from the 3' end region of the second strand.

38. The inhibitor according to any one of claims 35-37, wherein, The reverse nucleoside inter-bond is a 3'-3' reverse bond or a 5'-5' reverse bond.

39. The inhibitor according to any one of claims 35-38, wherein, One or more nucleosides on the first and / or second chains are modified to form modified nucleosides.

40. The inhibitor for the use according to claim 39, wherein, The modification is a modification of the 2'-OH group of the ribose, optionally selected from 2'-Me modification or 2'-F modification.

41. The inhibitor for the use according to claim 39 or 40, wherein, Starting from position 1 of the first chain, the first chain contains 2'-F modifications at any of the positions 14, 2, and 6, or any combination thereof.

42. The inhibitor according to any one of claims 39-41, wherein, Starting from position 1 of the second chain, the second chain includes 2'-F modifications at positions 7 and / or 9 and / or 11 and / or 13.

43. The inhibitor according to any one of claims 39-42, wherein, The first chain and the second chain contain 2'-Me and 2'-F modifications, respectively.

44. The inhibitor according to any one of claims 39-43, wherein the inhibitor is siRNA, The siRNA contains at least one thermally unstable modification, suitably located at one or more positions from position 1 to position 9 on the first strand, and / or at one or more positions on the second strand that match positions 1-9 on the first strand; wherein the unstable modification is selected from modified unlocking nucleic acids (UNA) and ethylene glycol nucleic acids (GNA), preferably ethylene glycol nucleic acids.

45. The inhibitor for the use according to claim 44, wherein, Starting from position 1 on the first strand, the siRNA contains at least one thermally unstable modification at position 7 on the first strand.

46. ​​The inhibitor according to any one of claims 39-45, wherein the inhibitor is siRNA, Starting from position 1 on the second strand, the siRNA contains three or more 2'-F modifications at positions 7 to 13 on the second strand, for example, four, five, six or seven 2'-F modifications at positions 7 to 13 on the second strand.

47. The inhibitor according to any one of claims 39-46, wherein the inhibitor is siRNA, Starting from position 1 of the second chain, the second chain contains at least three 2'-Me modifications, such as four, five, or six 2'-Me modifications, at positions 1 to 6 of the second chain.

48. The inhibitor according to any one of claims 39-47, wherein the inhibitor is siRNA, The first chain contains at least five consecutive 2'-Me modifications in the 3' end region; preferably, it includes the terminal nucleoside of the 3' end region, or at least a nucleoside within one or two nucleosides of the terminal nucleoside of the 3' end region.

49. The inhibitor according to any one of claims 39-48, wherein the inhibitor is siRNA, The first chain contains seven consecutive 2'-Me modifications in the 3' end region, preferably including terminal nucleosides in the 3' end region.

50. The inhibitor according to any one of claims 39-45, wherein the inhibitor is siRNA, The modified nucleoside of the second chain comprises any of the following modification patterns (5'-3'): (Me)8 – (F)3 – (Me) 10 。 51. The inhibitor according to any one of claims 39-45 or 50, wherein the inhibitor is siRNA, The first chain contains a 2' sugar modification pattern, wherein the modification is selected from at least 2'Me sugar modification and 2'F sugar modification, provided that the total number of 2'F sugar modifications in the first chain is not four or six.

52. The inhibitor according to any one of claims 39-45 or 50-51, wherein the inhibitor is siRNA, The modification is selected from at least 2'Me sugar modification and 2'F sugar modification, wherein the total number of 2'F sugar modifications in the first chain is three, five or seven.

53. The inhibitor for the use according to any one of claims 39-52, wherein, The siRNA oligomer also contains one or more thiophosphate nucleoside bonds.

54. The inhibitor for the use according to claim 53, wherein, The one or more thiophosphate nucleoside bonds are located between at least three consecutive positions in the 5' or 3' proximal region of the second chain, wherein the proximal region is preferably adjacent to the terminal region of the one or more of the base-free nucleosides of the second chain as defined in at least claim 35.

55. The inhibitor for the use according to claim 53 or 54, wherein, The one or more thiophosphate nucleoside bonds are located between at least three consecutive positions in the 5' end region and / or 3' end region of the first chain; wherein preferably, the end positions of the 5' end region and / or 3' end region of the first chain are connected to their adjacent positions by thiophosphate nucleoside bonds.

56. The inhibitor according to any one of claims 34-55, wherein, The oligomer is siRNA, and the second strand of the siRNA is directly or indirectly conjugated to one or more ligand moieties, wherein the ligand moieties are typically located in the terminal region of the second strand, preferably in its 3' terminal region.

57. The inhibitor for the use according to claim 56, wherein, The ligand portion comprises: i) one or more GalNAc ligands; and / or ii) One or more GalNAc ligand derivatives; and / or iii) One or more GalNAc ligands and / or GalNAc ligand derivatives conjugated to the siRNA via a linker.

58. The inhibitor for the use according to claim 57, wherein, The one or more GalNAc ligands and / or GalNAc ligand derivatives are conjugated directly or indirectly to the 5' or 3' end region of the second strand of the siRNA oligomer; preferably conjugated to its 3' end region.

59. The inhibitor for the use according to claim 57 or 58, wherein, The ligand portion includes 。 60. The inhibitor according to claim 57 or 58, having the following structure: ; in: R1 is selected independently from hydrogen, methyl, and ethyl each time it appears; R2 is selected from the following group: hydrogen, hydroxyl group, -OC 1-3 Alkyl group, -C(=O)OC 1-3 Alkyl, halogen, and nitro groups; X1 and X2 are each independently selected from methylene, oxygen, and sulfur when they appear; m is an integer from 1 to 6; n is an integer from 1 to 10; q, r, s, t, and v are each an independent integer between 0 and 4, provided that: (i) q and r cannot both be 0; and (ii) s, t and v cannot all be 0 at the same time; Z is an oligomer.

61. The inhibitor according to claim 57 or 58, having the following structure ; in: r and s are each independent integers from 1 to 16; and Z is an oligomer.

62. An inhibitor for use according to any one or more of the preceding claims, formulated as a pharmaceutical composition containing excipients and / or a carrier.

63. A method for preventing and / or treating and / or controlling insulin resistance, and / or elevated blood glucose levels, and / or elevated blood insulin levels, and / or elevated blood HbA1c levels, and / or elevated blood free fatty acid levels, and / or elevated blood fibrinogen levels, and / or elevated blood total cholesterol levels, and / or elevated blood LDL cholesterol levels, and / or elevated blood triglyceride levels, said method comprising administering an inhibitor of B4GALT1 expression and / or function to said patient.

64. A method for preventing and / or treating and / or controlling insulin resistance, and / or elevated blood glucose levels, and / or elevated blood insulin levels, and / or elevated blood HbA1c levels, and / or elevated blood free fatty acid levels, and / or elevated blood fibrinogen levels, and / or elevated blood total cholesterol levels, and / or elevated blood LDL cholesterol levels, and / or elevated blood triglyceride levels in patients who have or are at risk of developing metabolic and / or vascular diseases, said method comprising administering an inhibitor of B4GALT1 expression and / or function to said patient.

65. The method according to claim 63 or 64, wherein, The method can prevent and / or treat and / or control insulin resistance; preferably, the inhibitor can improve insulin resistance.

66. The method according to any one of claims 63-65, wherein, The method can prevent and / or treat and / or control elevated blood glucose levels; preferably, it is achieved by reducing elevated blood glucose levels.

67. The method according to any one of claims 63-66, wherein, The method can prevent and / or treat and / or control elevated blood insulin levels; preferably, it is achieved by reducing elevated blood insulin levels.

68. The method according to any one of claims 63-67, wherein, The method can prevent and / or treat and / or control elevated blood HbA1c levels; preferably, it is achieved by reducing elevated blood HbA1c levels.

69. The method according to any one of claims 63-68, wherein, The method can prevent and / or treat and / or control elevated blood free fatty acid levels; preferably, it is achieved by reducing elevated blood free fatty acid levels.

70. The method according to any one of claims 63-69, wherein, The method can prevent and / or treat and / or control elevated fibrinogen levels; preferably, it is achieved by reducing elevated fibrinogen levels.

71. The method according to any one of claims 63-70, wherein, The method can prevent and / or treat and / or control elevated total cholesterol levels; preferably, it is achieved by reducing elevated total cholesterol levels.

72. The method according to any one of claims 63-71, wherein, The method can prevent and / or treat and / or control elevated blood LDL cholesterol levels; preferably, it is achieved by reducing elevated blood LDL cholesterol levels.

73. The method according to any one of claims 63-72, wherein, The method can prevent and / or treat and / or control elevated blood triglyceride levels; preferably, it is achieved by reducing elevated blood triglyceride levels.

74. A method for preventing and / or treating and / or controlling vascular disease in a patient, said method comprising administering an inhibitor of B4GALT1 expression and / or function to said patient; wherein, The vascular disease is associated with insulin resistance, and / or elevated levels of free fatty acids, and / or elevated levels of fibrinogen, and / or elevated levels of total cholesterol, and / or elevated levels of LDL cholesterol, and / or elevated levels of triglycerides, and / or diabetes.

75. The method according to claim 74, wherein, The vascular disease is associated with insulin resistance; preferably, the inhibitor is able to improve insulin resistance.

76. The method according to claim 74 or 75, wherein, The vascular disease is associated with elevated levels of free fatty acids in the blood; preferably, the method is capable of reducing elevated levels of free fatty acids in the blood.

77. The method according to any one of claims 74-76, wherein, The vascular disease is associated with elevated fibrinogen levels; preferably, the method can reduce elevated fibrinogen levels.

78. The method according to any one of claims 74-77, wherein the vascular disease is associated with elevated total cholesterol levels; preferably, the method is capable of reducing elevated total cholesterol levels.

79. The method according to any one of claims 74-78, wherein, The vascular disease is associated with elevated blood LDL cholesterol levels; preferably, the method is capable of reducing elevated blood LDL cholesterol levels.

80. The method according to any one of claims 74-79, wherein the vascular disease is associated with elevated blood triglyceride levels; preferably, the method is capable of reducing elevated blood triglyceride levels.

81. The method according to any one of claims 74-80, wherein, The vascular disease is associated with diabetes; preferably, the inhibitor can improve diabetes.

82. A method for preventing and / or treating and / or controlling obesity, and / or weight gain, and / or metabolic syndrome in a patient, said method comprising administering an inhibitor of B4GALT1 expression and / or function to said patient.

83. The method according to claim 82, wherein, The obesity, and / or weight gain, and / or metabolic syndrome are associated with insulin resistance, and / or elevated blood free fatty acid levels; preferably, the method is capable of reducing elevated blood triglyceride levels.

84. The method of claim 83, wherein the obesity, and / or weight gain, and / or metabolic syndrome is associated with insulin resistance; preferably, the inhibitor is capable of improving insulin resistance.

85. The method according to claim 83 or 84, wherein, The obesity, and / or weight gain, and / or metabolic syndrome are associated with elevated blood free fatty acid levels; preferably, the method can reduce elevated blood free fatty acid levels.

86. The method according to any one of claims 83-85, wherein, The metabolic syndrome is further or independently associated with elevated total cholesterol levels, and / or elevated LDL cholesterol levels, and / or elevated triglyceride levels; preferably, the method is capable of reducing elevated total cholesterol levels, and / or elevated LDL cholesterol levels, and / or elevated triglyceride levels.

87. The method according to any one of claims 83-86, wherein, The metabolic syndrome is further or independently associated with elevated total blood cholesterol levels; preferably, the method is capable of reducing elevated total blood cholesterol levels.

88. The method according to any one of claims 83-87, wherein, The metabolic syndrome is further or independently associated with elevated blood LDL cholesterol levels; preferably, the method is capable of reducing elevated blood LDL cholesterol levels.

89. The method according to any one of claims 83-88, wherein, The metabolic syndrome is further or independently associated with elevated blood triglyceride levels; preferably, the method is capable of reducing elevated blood triglyceride levels.

90. The method according to any one of claims 63-89, wherein, The inhibitor suppresses the expression of B4GALT1 in hepatocytes.

91. The method according to any one of the preceding claims, wherein, The inhibitor of B4GALT1 expression and / or function is siRNA, particularly the siRNA defined in any one of claims 31-61.

92. The method according to any one of claims 63-91, wherein, The patient in question is a human patient.

93. Use of a nucleic acid or pharmaceutical composition in the prevention or treatment of vascular diseases, such as cardiovascular diseases.

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