Sirna for inhibiting angptl7, modified form thereof, and use thereof

Abstract

Disclosed are an siRNA for inhibiting ANGPTL7, a modified form thereof, and use thereof. The siRNAs all exhibit high inhibitory activity against ANGPTL7. Disclosed is a modified siRNA for inhibiting ANGPTL7, and the sequence of the siRNA exhibits good inhibitory efficiency.

Description

siRNAs and their modifications for inhibiting ANGPTL7 and their applications

[0001] Cross-reference to related applications

[0002] This application claims priority to Chinese Patent Application No. CN2025104620938, filed on April 14, 2025, entitled “siRNA for inhibiting ANGPTL7 and its modifications and applications”, the entire contents of which are incorporated herein by reference. Technical Field

[0003] This application relates to siRNA for inhibiting ANGPTL7 and its modifications and applications, belonging to the field of biotechnology. Background Technology

[0004] Glaucoma is a leading cause of vision loss. It is caused by damage to the optic nerve and loss of nerve fibers, and is usually associated with elevated intraocular pressure. Lowering intraocular pressure can reduce the development and progression of glaucoma and associated vision loss.

[0005] Human angiopoietin-like protein 7 (ANGPTL7), encoding angiopoietin-like protein 7 (ANGPTL6), is located on chromosome 1 at chromosomal region 1p36.22 and consists of six exons. ANGPTL7 belongs to the ANGPTL protein family and is expressed in sites such as the corneal stroma. Elevated levels of ANGPTL7 protein in the aqueous humor of glaucoma patients have been reported. Human genome analysis has shown that missense and nonsense variants of ANGPTL7 are associated with lower intraocular pressure and a lower risk of glaucoma. These findings suggest that interfering with ANGPTL7 may be a therapeutic strategy for glaucoma.

[0006] Small interfering RNA (siRNA) is an emerging and highly promising candidate for research. Based on RNA interference mechanisms, it can inhibit or block the expression of target genes of interest in a sequence-specific manner, thereby achieving the goal of treating diseases. Specifically, siRNA is loaded into an RNA-induced silencing complex.

[0007] RNA-induced silencing complex (RISC) involves a guide strand, also known as an antisense strand, that pairs complementaryly with the target nucleic acid in the mRNA of the target gene, causing the mRNA to degrade and thus inhibiting or blocking the expression of the target gene. Because siRNA recognizes the target RNA specifically through base pairing, thereby cleaving the target RNA and downregulating its level to inhibit the expression of the target gene, siRNA has a very broad range of applications. Summary of the Invention

[0008] To address the aforementioned problems, according to one aspect of this application, this application provides an siRNA for inhibiting ANGPTL7, said siRNA containing a sense strand and an antisense strand, said sense strand and antisense strand being at least partially anti-complementary to form a double-stranded region; said sense strand of said siRNA comprises a nucleic acid molecule with a nucleotide sequence as shown in any one of SEQ ID NO. 1 to 255; said antisense strand of said siRNA comprises a nucleic acid molecule with a nucleotide sequence as shown in any one of SEQ ID NO. 256 to 510.

[0009] According to another aspect of this application, this application provides an siRNA for inhibiting ANGPTL7, the siRNA containing a sense strand and an antisense strand, the sense strand and the antisense strand being at least partially anticomplementary to form a double-stranded region; the sense strand of the siRNA comprises at least 15 consecutive nucleotides with nucleotide sequences as shown in any one of the nucleic acid molecules or nucleic acid sequences shown in any one of SEQ ID NO. 1 to 255, differing by no more than 3 nucleotides; the antisense strand of the siRNA comprises at least 15 consecutive nucleotides with nucleotide sequences as shown in any one of the nucleic acid molecules or nucleic acid sequences shown in any one of SEQ ID NO. 256 to 510, differing by no more than 3 nucleotides.

[0010] Furthermore, at least one nucleotide in the sense or antisense strand of the siRNA is a modified nucleotide.

[0011] Furthermore, the modified nucleotide is a compound formed by modifying the ribose of the nucleotide, or a compound formed by modifying the backbone between nucleotides, or a compound formed by modifying the bases on the nucleotide, or a compound formed by modifying the ends of the siRNA.

[0012] Further, the modified nucleotide is selected from at least one of the following: 2'-methoxy modified nucleotides, 2'-fluoro modified nucleotides, phosphate thioester modified nucleotides, deoxy-nucleotides, 2'-deoxy-nucleotides, 3'-terminal deoxy-thymidine (dT) nucleotides, locked nucleotides, non-locked nucleotides, configuration-restricted nucleotides, restricted ethyl nucleotides, base-free nucleotides, reverse nucleotides, reverse base-free nucleotides, 2'-amino-modified nucleotides, 2'-O-allyl-modified nucleotides, 2'-C-alkyl-modified nucleotides, 2'-hydroxy Nucleotides modified with α-, 2'-methoxyethyl, 2'-O-alkyl, morpholino, aminophosphate, nucleotides containing non-natural bases, tetrahydropyran, 1,5-dehydrohexyl, cyclohexenyl, glycol-nucleotides (GNA), uncyclic nucleotides (UNA), nucleotides containing methylphosphonate, nucleotides containing 5'-phosphate, nucleotides containing 5'-phosphate mimics, or terminal nucleotides linked to cholesterol derivatives or dodecanoic acid bisdecamide groups.

[0013] Alternatively, the modified nucleotides include methoxylated nucleotides, fluorinated nucleotides, phosphate-thioester linked nucleotides, deoxyribonucleotide-substituted nucleotides, 2'-deoxy-nucleotides, 3'-terminal deoxy-thymidine nucleotides, 3'-deoxy-nucleotides, inosine nucleotides, locked nucleotides, non-locked nucleotides, configuration-restricted nucleotides, restricted ethyl nucleotides, baseless nucleotides, reverse nucleotides, reverse baseless nucleotides, vinyl phosphate-modified nucleotides, 2'-amino-modified nucleotides, 2'-O-allyl-modified nucleotides, 2'-C-alkyl-modified nucleotides, 2'- The nucleotides are: hydroxyl-modified nucleotides, 2'-methoxyethyl-modified nucleotides, 2'-O-alkyl-modified nucleotides, morpholinonucleotide-modified nucleotides, aminophosphate-modified nucleotides, nucleotides containing non-natural bases, tetrahydropyran-modified nucleotides, 1,5-dehydrohexylol-modified nucleotides, cyclohexenyl-modified nucleotides, ethylene glycol nucleic acid-modified nucleotides, open-ring nucleotide-modified nucleotides, nucleotides containing methylphosphate groups, nucleotides containing 5'-phosphate groups, nucleotides containing 5'-phosphate analogs, terminal nucleotides linked to cholesterol derivatives, and terminal nucleotides linked to dodecanoic acid bisdecamide groups.

[0014] Furthermore, the fluorinated nucleotides are located in both the antisense and sense strands of the nucleotide sequence, and, following the direction from the 5' end to the 3' end, the nucleotides at positions 7, 8, and 9, or positions 5, 7, 8, and 9, or positions 9, 10, and 11, or positions 7, 9, 10, and 11 of the sense strand are fluorinated nucleotides, while the nucleotide groups at the remaining positions in the sense strand are each non-fluorinated nucleotide groups; the nucleotides at positions 2, 6, 12, 14, and 16, or positions 2, 8, 12, 14, and 16, or positions 2, 9, 12, 14, and 16, or positions 2, 10, and 12 of the antisense strand are fluorinated nucleotides. The nucleotides at positions 14 and 16, or positions 2, 6, 10, 12, and 14, or positions 2, 8, 10, 14, and 16, or positions 2, 6, 10, 14, and 16, or positions 2, 4, 12, 14, and 16, or positions 2, 6, 14, and 16, or positions 2, 10, 12, 14, 16, 18, and 20, or positions 2, 6, 8, 14, and 16, or positions 14, or positions 10, 12, 14, 16, 18, and 20, or positions 2, 14, and 16 are fluorinated nucleotides, and the nucleotide groups at the remaining positions in the antisense strand are each non-fluorinated nucleotide groups;

[0015] Each of the non-fluorinated modified nucleotide groups is independently selected from one of the nucleotide groups or nucleotide analog groups formed by replacing the hydroxyl group at the 2' position of the ribosyl group with a non-fluorinated group.

[0016] Alternatively, in the direction from the 5' end to the 3' end, at least one nucleotide in positions 2 to 18 of the antisense strand is a thermally unstable modified nucleotide.

[0017] Alternatively, the fluorinated nucleotides are located in both the antisense and sense strands of the nucleotide sequence, and, in the direction from the 5' end to the 3' end, the nucleotides at positions 7, 8, and 9, or positions 5, 7, 8, and 9, or positions 9, 10, and 11, or positions 7, 9, 10, and 11, of the sense strand are fluorinated nucleotides, while the nucleotide groups at the remaining positions in the sense strand are each non-fluorinated nucleotide groups; the nucleotides at positions 2, 6, 12, 14, and 16, or positions 2, 8, 12, 14, and 16, or positions 2, 9, 12, 14, and 16, of the antisense strand are fluorinated nucleotides. 4. The nucleotides at positions 16, 2, 6, 10, 12, 14, 2, 8, 10, 14, 16, 2, 6, 10, 14, 16, 2, 4, 12, 14, 16, 2, 6, 14, 16, 2, 10, 12, 14, 16, 18, 20, 2, 6, 8, 14, 16, 14, 10, 12, 14, 16, 18, 20, 2, 14, 16 are fluorinated nucleotides, and the nucleotide groups at the remaining positions in the antisense strand are each non-fluorinated nucleotide groups.

[0018] Each of the non-fluorinated modified nucleotide groups is independently selected from one of the nucleotide groups or nucleotide analog groups formed by replacing the 2' hydroxyl group of the ribosyl group with a non-fluorinated group, wherein the non-fluorinated modified nucleotide is a methoxy-modified nucleotide and / or a 2′-deoxynucleotide and / or an inosine nucleotide and / or a ethylene glycol-modified nucleotide.

[0019] Furthermore, in the direction from the 5' end to the 3' end, at least one nucleotide in the 5th, 7th, 10th, 11th, 12th, 13th, and 16th positions of the antisense strand is a thermally unstable modified nucleotide.

[0020] Alternatively, in the direction from the 5' end to the 3' end, at least one nucleotide in positions 2 to 18 of the antisense strand is a heat-labile modified nucleotide; optionally, in the direction from the 5' end to the 3' end, at least one nucleotide in positions 5, 7, 10, 11, 12, 13, and 16 of the antisense strand is a heat-labile modified nucleotide.

[0021] Furthermore, in the direction from the 5' end to the 3' end, at least two nucleotides in the antisense strand 2 to 8 are thermally unstable modified nucleotides.

[0022] Furthermore, the thermally unstable modified nucleotide is ethylene glycol nucleic acid nucleotide (GNA) and / or 2′-deoxynucleotide.

[0023] Furthermore, the non-fluorinated modified nucleotide group is a methoxylated modified nucleotide.

[0024] Furthermore, the nucleotides linked by the thiophosphate group are located in the antisense and sense strands of the nucleotide sequence, and, in the direction from the 5' end to the 3' end, at least the nucleotides at positions 1 and 2, 2 and 3, 19 and 20, and 20 and 21 of the antisense strand are linked by the thiophosphate group.

[0025] Alternatively, in the direction from the 5' end to the 3' end, at least the first and second nucleotides of the positive strand are linked by a phosphate thioester group;

[0026] Alternatively, in the direction from the 5' end to the 3' end, at least the nucleotides at positions 1 and 2, and positions 2 and 3 of the sense strand are linked by thiophosphate groups, and at least the nucleotides at positions 1 and 2, positions 2 and 3, positions 19 and 20, and positions 20 and 21 of the antisense strand are linked by thiophosphate groups.

[0027] Alternatively, in the direction from the 5' end to the 3' end, at least the first and second positions of the sense strand are linked by a thiophosphate group, and at least the first and second, second and third, 19th and 20th, and 20th and 21st nucleotides of the antisense strand are linked by a thiophosphate group.

[0028] Alternatively, in the direction from the 5' end to the 3' end, at least the first and second positions, and the 20th and 21st positions of the sense strand are linked by thiophosphate groups, and at least the first and second positions, the second and third positions, the 19th and 20th positions, and the 20th and 21st positions of the antisense strand are linked by thiophosphate groups.

[0029] Alternatively, in the direction from the 5' end to the 3' end, at least the nucleotides at positions 1 and 2, 2 and 3, 19 and 20, and 20 and 21 of the sense strand are linked by thiophosphate groups, and at least the nucleotides at positions 1 and 2, 2 and 3, 19 and 20, and 20 and 21 of the antisense strand are linked by thiophosphate groups.

[0030] Alternatively, in the direction from the 5' end to the 3' end, the nucleotides linked by the thiophosphate group are located in the antisense and sense strands of the nucleotide sequence, and in the direction from the 5' end to the 3' end, at least the nucleotides at positions 1 and 2, 19 and 20, and 20 and 21 of the sense strand are linked by thiophosphate groups, and at least the nucleotides at positions 1 and 2, 2 and 3, 19 and 20, and 20 and 21 of the antisense strand are linked by thiophosphate groups.

[0031] Alternatively, the nucleotides linked by the thiophosphate group are located in the antisense and sense strands of the nucleotide sequence, and, in the direction from the 5' end to the 3' end, at least the nucleotides at the 1st and 2nd, 2nd and 3rd, penultimate and penultimate, and penultimate and penultimate positions of the antisense strand are linked by the thiophosphate group.

[0032] Alternatively, in the direction from the 5' end to the 3' end, at least the first and second nucleotides of the positive strand are linked by a phosphate thioester group;

[0033] Alternatively, in the direction from the 5' end to the 3' end, at least the nucleotides at positions 1 and 2, and positions 2 and 3 of the sense strand are linked by thiophosphate groups, and at least the nucleotides at positions 1 and 2, positions 2 and 3, positions 1 to 1 and 2 to 1, and positions 2 to 1 to 3 to 1 of the antisense strand are linked by thiophosphate groups.

[0034] Alternatively, in the direction from the 5' end to the 3' end, at least the first and second positions of the sense strand are linked by a thiophosphate group, and at least the first and second, second and third, penultimate and penultimate, and penultimate and penultimate nucleotides of the antisense strand are linked by a thiophosphate group.

[0035] Alternatively, in the direction from the 5' end to the 3' end, at least the first and second positions, and the 20th and 21st positions of the sense strand are linked by thiophosphate groups, and at least the first and second positions, the second and third positions, the penultimate and penultimate positions, and the penultimate and penultimate positions of the antisense strand are linked by thiophosphate groups.

[0036] Alternatively, in the direction from the 5' end to the 3' end, at least the nucleotides at positions 1 and 2, 2 and 3, penultimate and penultimate, and penultimate and penultimate of the sense strand are linked by thiophosphate groups, and at least the nucleotides at positions 1 and 2, 2 and 3, penultimate and penultimate, and penultimate and penultimate of the antisense strand are linked by thiophosphate groups.

[0037] Alternatively, in the direction from the 5' end to the 3' end, the nucleotides linked by the thiophosphate group are located in the antisense and sense strands of the nucleotide sequence, and in the direction from the 5' end to the 3' end, at least the nucleotides at positions 1 and 2, 19 and 20, and 20 and 21 of the sense strand are linked by thiophosphate groups, and at least the nucleotides at positions 1 and 2, 2 and 3, penultimate and penultimate, and penultimate and penultimate of the antisense strand are linked by thiophosphate groups.

[0038] Furthermore, the 5' and / or 3' ends of the siRNA's sense and / or antisense strands contain at least one reverse-abase-free nucleotide.

[0039] Furthermore, the first nucleotide at the 5' and / or 3' end of the siRNA's sense and / or antisense strands is a vinyl phosphate-modified nucleotide.

[0040] Furthermore, the siRNA is coupled to or conjugated with one or more nucleotides on at least one strand via a linker or vector, or with one or more lipophilic moieties.

[0041] Furthermore, the siRNA has a ligand coupled to the ends of its sense and / or antisense strands, wherein the ligand is GalNAc.

[0042] This application also provides a product for inhibiting ANGPTL7, characterized in that the product comprises an active ingredient and a pharmaceutically acceptable carrier, wherein the active ingredient is siRNA as described in any of the above claims.

[0043] This application also provides the use of siRNA or the above-described product according to any one of the foregoing in the preparation of products for the prevention, diagnosis and / or treatment of pathological conditions or diseases caused by ANGPTL7.

[0044] In one embodiment of this application, taking the positive and antisense strands shown in SEQ ID NO.1 and SEQ ID NO.256 as examples, the positive strand comprises: a nucleotide sequence as shown in SEQ ID NO.1 or a nucleotide sequence having at least 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% identity with the nucleotide sequence shown in SEQ ID NO.1 and retaining the biological function of its derived sequence; the antisense strand comprises: a nucleotide sequence as shown in SEQ ID NO.256 or a nucleotide sequence having at least 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% identity with the nucleotide sequence shown in SEQ ID NO.256 and retaining the biological function of its derived sequence. The remaining positive and antisense strands each have at least 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% identity and retain the biological function of their derived sequences.

[0045] In one embodiment of this application, the 5' end of the siRNA sense strand may also be linked to 1-3 nucleotides complementary to the 3' end of the antisense strand;

[0046] Alternatively, the 3' end of the siRNA sense strand may also be linked to 1-3 nucleotides complementary to the 5' end of the antisense strand;

[0047] Alternatively, the 5' end of the siRNA antisense strand may also be linked to 1-3 nucleotides complementary to the 3' end of the sense strand;

[0048] Alternatively, the 3' end of the siRNA antisense strand may be linked to 1-3 nucleotides complementary to the 5' end of the sense strand.

[0049] In one embodiment of this application, the 5' end of the siRNA sense strand may also be linked to two nucleotides complementary to the 3' end of the antisense strand;

[0050] Alternatively, the 3' end of the siRNA sense strand may also be linked to two nucleotides complementary to the 5' end of the antisense strand;

[0051] Alternatively, the 5' end of the siRNA antisense strand may also be linked to two nucleotides complementary to the 3' end of the sense strand;

[0052] Alternatively, the 3' end of the siRNA antisense strand may also be linked to two nucleotides complementary to the 5' end of the sense strand.

[0053] In one embodiment of this application, the sense strand and antisense strand of the double-stranded RNA molecule are complementary to form the double-stranded region of the siRNA, the 3' end of the sense strand forms a blunt end, and the 3' end of the antisense strand has 1-3 protruding nucleotides extending out of the double-stranded region;

[0054] Alternatively, the sense and antisense strands of the double-stranded RNA molecule are complementary to form the double-stranded region of the siRNA, the 5' end of the sense strand forms a blunt end, and the 5' end of the antisense strand has 1-3 protruding nucleotides extending out of the double-stranded region;

[0055] Alternatively, the sense and antisense strands of the double-stranded RNA molecule are complementary to form the double-stranded region of the siRNA, with the 3' end of the sense strand forming a blunt end and the 3' end of the antisense strand forming a blunt end;

[0056] Alternatively, the sense and antisense strands of the double-stranded RNA molecule are complementary to form the double-stranded region of the siRNA, and the 3' ends of the sense and antisense strands have 1-3 protruding nucleotides extending out of the double-stranded region, or the 5' ends of the sense and antisense strands have 1-3 protruding nucleotides extending out of the double-stranded region.

[0057] In one embodiment of this application, the siRNA is prepared by solid-phase synthesis or liquid-phase synthesis.

[0058] In one embodiment of this application, the nucleotides in the siRNA are each independently modified or unmodified nucleotides.

[0059] In one embodiment of this application, each nucleotide in the siRNA is an unmodified nucleotide.

[0060] In one embodiment of this application, some or all of the nucleotides in the siRNA are modified nucleotides, and these modifications on the nucleotide groups do not cause the siRNA of this disclosure to significantly weaken or lose its function of inhibiting ANGPTL7 gene expression.

[0061] In one embodiment of this application, at least one nucleotide in the sense or antisense strand of the siRNA is a modified nucleotide.

[0062] In one embodiment of this application, all nucleotides in the sense and / or antisense strands of the siRNA are modified nucleotides, and these modifications on the nucleotide groups do not cause a significant weakening or loss of the function of the siRNA disclosed herein in inhibiting ANGPTL7 gene expression.

[0063] In one embodiment of this application, each nucleotide in the sense and antisense strands of the siRNA is independently a fluorinated nucleotide or a non-fluorinated nucleotide.

[0064] In one embodiment of this application, the fluorinated nucleotide is located in both the antisense and sense strands of the nucleotide sequence, and the fluorinated nucleotide is present at least at one of the following positions:

[0065] Following the direction from the 5' end to the 3' end, at least the 7th, 8th, and 9th nucleotides of the positive strand are fluorinated nucleotides; at least the 5th, 7th, 8th, and 9th nucleotides of the positive strand are fluorinated nucleotides; at least the 9th, 10th, and 11th nucleotides of the positive strand are fluorinated nucleotides; at least the 7th, 9th, 10th, and 11th nucleotides of the positive strand are fluorinated nucleotides.

[0066] Following the direction from the 5' end to the 3' end, at least the 2nd, 6th, 12th, 14th, and 16th nucleotides of the antisense strand are fluorinated nucleotides; at least the 2nd, 8th, 12th, 14th, and 16th nucleotides of the antisense strand are fluorinated nucleotides; at least the 2nd, 9th, 12th, 14th, and 16th nucleotides of the antisense strand are fluorinated nucleotides; at least the 2nd, 10th, 12th, 14th, and 16th nucleotides of the antisense strand are fluorinated nucleotides; at least the 2nd, 6th, 10th, 12th, and 14th nucleotides of the antisense strand are fluorinated nucleotides; at least the 2nd, 8th, 10th, 14th, and 16th nucleotides of the antisense strand are fluorinated nucleotides; at least the 2nd, 6th, 10th, 14th, and 16th nucleotides of the antisense strand are fluorinated nucleotides; at least the 2nd, 6th, 10th, 14th, and 16th nucleotides of the antisense strand are nucleotides. The acid is a fluorinated nucleotide; at least the 2nd, 4th, 12th, 14th, and 16th nucleotides of the antisense strand are fluorinated nucleotides; at least the 2nd, 6th, 14th, and 16th nucleotides of the antisense strand are fluorinated nucleotides; at least the 2nd, 10th, 12th, 14th, 16th, 18th, and 20th nucleotides of the antisense strand are fluorinated nucleotides; at least the 2nd, 6th, 8th, 14th, and 16th nucleotides of the antisense strand are fluorinated nucleotides; at least the 14th nucleotide of the antisense strand is a fluorinated nucleotide; at least the 10th, 12th, 14th, 16th, 18th, and 20th nucleotides of the antisense strand are fluorinated nucleotides; at least the 2nd, 14th, and 16th nucleotides of the antisense strand are fluorinated nucleotides.

[0067] Alternatively, any combination thereof, meaning that the fluorinated nucleotides of the sense and antisense strands are selected from any combination thereof. The remaining nucleotide groups in the sense / antisense strands are each non-fluorinated nucleotide groups.

[0068] In one embodiment of this application, each of the non-fluorinated modified nucleotide groups is independently selected from one of the nucleotide groups or nucleotide analog groups formed by replacing the hydroxyl group at the 2′ position of the ribosome of the nucleotide group with a non-fluorinated group.

[0069] In one embodiment of this application, in the direction from the 5' end to the 3' end, at least one nucleotide of the antisense strand is a thermally unstable modified nucleotide, and each of the remaining non-fluorinated modified nucleotide groups is independently selected from a nucleotide group or a nucleotide analog group formed by replacing the hydroxyl group at the 2' position of the ribosyl group with a non-fluorinated group.

[0070] In one embodiment of this application, at least one nucleotide from 2 to 8 of the antisense strand is a thermally unstable modified nucleotide, following the direction from the 5' end to the 3' end.

[0071] In one embodiment of this application, at least two nucleotides from 2 to 8 of the antisense strand are thermally unstable modified nucleotides, arranged in the direction from the 5' end to the 3' end.

[0072] In one embodiment of this application, the nucleotides at least the 5th and 7th positions of the antisense strand are thermally unstable modified nucleotides, following the direction from the 5' end to the 3' end.

[0073] In one embodiment of this application, "fluorinated nucleotide" refers to a nucleotide formed by replacing the hydroxyl group at the 2' position of the ribosyl group with fluorine, having the structure shown in formula (1). The non-fluorinated nucleotide is independently selected from nucleotides or nucleotide analogs formed by replacing the hydroxyl group at the 2' position of the ribosyl group with a non-fluorinated group.

[0074] In one embodiment of this application, the nucleotide formed by replacing the hydroxyl group at the 2' position of the ribosome with a non-fluorinated group is well known to those skilled in the art. These nucleotides may be selected from one of the following: 2'-alkoxy modified nucleotides, 2′-substituted alkoxy modified nucleotides, 2′-alkyl modified nucleotides, 2′-substituted alkyl modified nucleotides, 2′-amino modified nucleotides, 2′-substituted amino modified nucleotides, and 2′-deoxynucleotides.

[0075] In one embodiment of this application, the 2′-alkoxy modified nucleotide is a 2′-methoxy (2′-OMe) modified nucleotide, as shown in formula (2), i.e., methoxy modified; the 2′-substituted alkoxy modified nucleotide may be, for example, a 2′-O-methoxyethyl (2′-MOE) modified nucleotide, as shown in formula (3), a 2′-amino (2′-NH2) modified nucleotide as shown in formula (4), and a 2′-deoxynucleotide (DNA) as shown in formula (5), wherein Base refers to the modified or unmodified nucleotide base A, U, G, C, T or other nucleotide bases:

[0076] In one embodiment of this application, the nucleotide analogue refers to a group that can replace a nucleotide in nucleic acid, but whose structure is different from that of adenine ribonucleotide, guanine ribonucleotide, cytosine ribonucleotide, uracil ribonucleotide, or thymine deoxyribonucleotide.

[0077] In one embodiment of this application, the nucleotide analog may be an isonucleotide, a bridged nucleotide, or an acyclic nucleotide.

[0078] In one embodiment of this application, the bridged nucleic acid (BNA) refers to a restricted or inaccessible nucleotide. The BNA may contain a bridging structure with a "fixed" C3'-endoglucan condensation, such as a five-membered ring, a six-membered ring, or a seven-membered ring. Typically, the bridge is incorporated into the 2'-, 4'-position of the ribose to provide a 2',4'-BNA nucleotide.

[0079] In one embodiment of this application, the BNA may be LNA, ENA, cET BNA, etc., wherein LNA is as shown in formula (6), ENA is as shown in formula (7), and cET BNA is as shown in formula (8), wherein Base refers to the modified or unmodified nucleotide bases A, U, G, C, T or other nucleotide bases:

[0080] In one embodiment of this application, the thermally unstable modification may include, but is not limited to, debasement modification; mismatch with relative nucleotides in the opposite chain; and sugar modification, such as 2'-deoxy modification or acyclic nucleotides, for example, unlocked nucleic acid (UNA) or glycol nucleic acid (GNA).

[0081] Examples of debasing modifications include, but are not limited to, the following, with structural formulas as shown in formulas (9) to (15):

[0082] Where R = H, Me, Et or OMe; R' = H, Me, Et or OMe; R” = H, Me, Et or OMe.

[0083] In one embodiment of this application, sugar modification includes, but is not limited to, the following: 2'-deoxynucleotides, unlocked nucleic acids, and glycol-based nucleic acids.

[0084] In one embodiment of this application, an acyclic nucleotide refers to any nucleotide having a noncyclic ribose, for example, wherein any bond between ribose carbons in the nucleotide (e.g., C1'-C2', C2'-C3', C3'-C4', C4'-O4', or C1'-O4') is absent and / or at least one of the ribose carbons or oxygen (e.g., C1', C2', C3', C4', or O4') is absent independently or in combination.

[0085] In one embodiment of this application, UNA represents an unlocked acyclic nucleic acid (or open-ring nucleotide) in which any bond of the sugar is removed, forming an unlocked "sugar" residue. In one instance, UNA also encompasses monomers in which the bond between C1' and C4' has been removed (i.e., the covalent carbon-oxygen-carbon bond between C1' and C4' carbons). In another instance, the C2'-C3' bond of the sugar (i.e., the covalent carbon-carbon bond between C2' and C3' carbons) is removed (see Mikhailov et al., Tetrahedron Letters, 26(17):2059(1985); and Fluiter et al., Mol. Biosyst., 10:1039(2009), which are incorporated herein by reference in their entirety). Acyclic derivatives provide greater skeletal flexibility without affecting Watson-Crick pairing. Acyclic nucleotides can be linked via 2'-5' or 3'-5' bonds.

[0086] In one embodiment of this application, GNA represents diol nucleic acid, which is a polymer similar to DNA or RNA, but with a different "backbone" composed of repeating glycerol units linked by phosphodiester bonds. The structure of A (GNA) is shown in Formula (16), the structure of G (GNA) is shown in Formula (17), the structure of C (GNA) is shown in Formula (18), the structure of U (GNA) is shown in Formula (19), and the structure of T (GNA) is shown in Formula (20).

[0087] In one embodiment of this application, inosine nucleotides are also thermally unstable modifications. The inosine nucleotides are represented by the following structures: dI structure as shown in formula (21), rI structure as shown in formula (22), 2'-F rI structure as shown in formula (23), and 2'OMe rI structure as shown in formula (24).

[0088] Thermally unstable modifications to the duplex can be mismatches (i.e., non-complementary base pairs) between thermally unstable nucleotides and relative nucleotides in the opposing strands of the dsRNA duplex. Exemplary mismatched base pairs include G:G, G:A, G:U, G:T, A:A, A:C, C:C, C:U, C:T, U:U, T:T, U:T, or combinations thereof. Other mismatched base pairings known in the art are also applicable to this application. Mismatches can occur between nucleotides, which are naturally occurring nucleotides or modified nucleotides; that is, mismatched base pairs can occur between nucleobases from their respective nucleotides, regardless of modifications to the ribose of the nucleotides. In some embodiments, the dsRNA molecule contains at least one nucleobase in the mismatched pair that is a 2'-deoxynucleobase; for example, a 2'-deoxynucleobase in the sense strand.

[0089] Further examples of thermally unstable modified nucleotides are described in detail in WO2018098328A1 (the entire contents of which are incorporated herein by reference), and thermally unstable modifications in this application are equivalent to thermally destabilizing modifications in WO2018098328A1.

[0090] In one embodiment of this application, the nucleotides at least the 5th and 7th positions of the antisense strand are thermally unstable modified nucleotides, in the direction from the 5' end to the 3' end; the thermally unstable modified nucleotides are ethylene glycol nucleic acid nucleotides (GNA) and / or 2′-deoxynucleotides, wherein the 2′-deoxynucleotides are located in the antisense strand of the nucleotide sequence.

[0091] In one embodiment of this application, at least one phosphate group in the sense or antisense strand of the siRNA is a phosphate group with a modifying group.

[0092] In one embodiment of this application, at least a portion of the phosphate ester groups and / or ribosomes in the phosphate-sugar backbone of at least one single strand of the sense and antisense strands of the siRNA are phosphate ester groups and / or ribosomes with modifying groups.

[0093] In one embodiment of this application, the phosphate ester group with the modifying group is a thiophosphate ester group formed by replacing at least one oxygen atom in the phosphate diester bond of the phosphate ester group with a sulfur atom.

[0094] In one embodiment of this application, the phosphate group with the modifying group is a thiophosphate group having the structure shown in formula (25). In one embodiment of this application, the nucleotide linked to the thiophosphate group is shown in formula (26). In one embodiment of this application, the VP-modified nucleotide is a vinyl phosphate modified nucleotide. In one embodiment of this application, the nucleotide modified by VP and methoxy groups, i.e., the nucleotide modified by 5'-(E)-vinyl-2'-methoxy phosphonate group (5'-(E)-VP-2'-OMe), is shown in formula (27). In one embodiment of this application, the nucleotide modified by VP, methoxy groups, and thiophosphate groups, i.e., the nucleotide modified by 5'-PS (i.e., the nucleotide modified by 5'-(E)-vinyl-2'-methoxy thiophosphonate group), is shown in formula (28).

[0095] In one embodiment of this application, the thiophosphate group linkage is present at least at one of the following positions: between the first and second nucleotides at either end of the sense or antisense strand; between the second and third nucleotides at either end of the sense or antisense strand; or any combination thereof.

[0096] In one embodiment of this application, the thiophosphate group linkage is present at all of the above-mentioned positions except for the end of the positive chain 5'.

[0097] In one embodiment of this application, the thiophosphate group linkage is present at all of the above-mentioned positions except for the end of the positive chain 3'.

[0098] In one embodiment of this application, the thiophosphate group linkage is present at at least one of the following locations:

[0099] Between the first and second nucleotides at the 5' end of the positive strand;

[0100] Between the second and third nucleotides at the 5' end of the positive strand;

[0101] Between the first and second nucleotides at the 3' end of the positive strand;

[0102] Between the second and third nucleotides at the 3' end of the positive strand;

[0103] Between the first and second nucleotides at the 5' end of the antisense strand;

[0104] Between the second and third nucleotides at the 5' end of the antisense strand;

[0105] Between the first and second nucleotides at the 3' end of the antisense strand; and

[0106] Between the second and third nucleotides at the 3' end of the antisense strand.

[0107] In one embodiment of this application, the reverse nucleotide represents the following structure, as shown in formulas (29) to (32).

[0108] In one embodiment of this application, invAb represents a reverse abase-free nucleotide, and the structural formula of invAb is shown in formula (33).

[0109] In one embodiment of this application, the 5' end and / or 3' end of the siRNA sense strand or antisense strand contains at least one reverse abase-free nucleotide.

[0110] In one embodiment of this application, the 5' end and / or 3' end of the siRNA positive strand contains at least one reverse abase-free nucleotide.

[0111] In one embodiment of this application, the siRNA is coupled to a functional molecule, including N-acetylgalactosamine (GalNAc), lipophilic molecules, peptides, small molecule drugs, antibodies, etc.

[0112] In one embodiment of this application, siRNA may be coupled to one or more coupling groups containing functional molecules.

[0113] In one embodiment of this application, the pharmaceutically acceptable targeting group in the siRNA conjugate is galactose or N-acetylgalgactosamine (GalNAc). N-acetylgalgactosamine (GalNAc) is a ligand that binds to the asialoglycoprotein receptor (ASGPR) on the liver surface. The ASGPR is an endocytic receptor specifically expressed by hepatocytes. N-acetylgalgactosamine serves as a targeting molecule to deliver small RNA to the liver.

[0114] In one embodiment of this application, the galactose or N-acetylgalactosamine molecule can be monovalent, divalent, trivalent, or tetravalent; the monovalent, divalent, trivalent, and tetravalent respectively refer to the molar ratio of siRNA molecule to galactose or N-acetylgalactosamine molecule in the siRNA conjugate formed by the siRNA molecule and the coupling group containing the galactose or N-acetylgalactosamine molecule as the target group being 1:1, 1:2, 1:3, or 1:4.

[0115] In one embodiment of this application, when siRNA is coupled to a coupling group containing N-acetylgalactosamine, the N-acetylgalactosamine molecule is trivalent or tetravalent.

[0116] In one embodiment of this application, when siRNA is coupled to a coupling group containing N-acetylgalactosamine, the N-acetylgalactosamine molecule is trivalent.

[0117] In one embodiment of this application, siRNA may be coupled to one or more coupling groups containing N-acetylgalactosamine.

[0118] In one embodiment of this application, the targeting group can be linked to the siRNA molecule via a suitable adapter, and those skilled in the art can select a suitable adapter according to the specific type of the targeting group.

[0119] The types of adapters, targeting groups, and the methods of ligation with siRNA are described in detail in WO2015006740A2 (the entire contents of which are incorporated herein by reference).

[0120] In one embodiment of this application, the 5' and / or 3' ends of the sense or antisense strand of the siRNA are coupled with a ligand, which may be a lipophilic moiety or GalNAc.

[0121] In one embodiment of this application, the siRNA has a ligand coupled to the 3' end of the positive strand, the ligand being GalNAc.

[0122] In one embodiment of this application, the 3' end of the positive strand of the siRNA is connected to the ligand via a thiophosphate group.

[0123] In one embodiment of this application, the siRNA conjugate formed by GalNAc and siRNA molecules has the structure shown in formula (34) below:

[0124] The targeting group is a small molecule, peptide, aptamer, or antibody that targets integrin.

[0125] The targeting group is a small molecule, polypeptide, aptamer, or antibody that targets integrin avβ6.

[0126] The types of adapters, targeting groups, and the methods of linking to siRNA are described in detail in WO2022056286A1, WO2024129931A1, WO2025067433A1, and WO2025067423A1 (the entire contents of which are incorporated herein by reference).

[0127] In one embodiment of this application, the siRNA conjugate formed by the ligand targeting integrin avβ6 and the siRNA molecule has the structure shown in the following formula:

[0128] In one embodiment of this application, the functional molecule is a lipophilic molecule (or a lipophilic or lipophilic moiety). The siRNA comprises one or more lipophilic moieties coupled or conjugated to one or more nucleotides on at least one chain via a linker or vector. The lipophilic moieties include, but are not limited to, one or more of saturated alkanes, unsaturated alkanes, saturated fatty acids, unsaturated fatty acids, and cholesterol of varying chain lengths.

[0129] In one embodiment of this application, the lipophilic moiety is aliphatic, cyclic (e.g., alicyclic), or polycyclic, such as alicyclic compounds, such as steroids (e.g., sterols), or straight-chain or branched aliphatic hydrocarbons. Exemplary lipophilic moieties include lipids, cholesterol, retinoic acid, cholic acid, adamantaneacetic acid, 1-pyrenebutyric acid, dihydrotestosterone, 1,3-bis-O-(hexadecyl)glycerol, geraniol, hexadecylglycerol, borneol, menthol, 1,3-propanediol, heptadecanyl, palmitic acid, myristic acid, O3-(oleoyl)lithocholic acid, O3-(oleoyl)cholenic acid, ibuprofen, naproxen, dimethoxytriphenylmethyl, or phenoxazine.

[0130] In one embodiment of this application, the lipophilic portion also includes saturated or unsaturated C4-C. 30 Hydrocarbon chains (e.g., C4-C) 30 A lipophilic moiety comprising an alkyl or alkenyl group and optional functional groups selected from the group consisting of hydroxyl, amine, carboxylic acid, sulfonate, phosphate, thiol, azide, and alkyne. These functional groups can be used to attach the lipophilic moiety to an iRNA agent. In some embodiments, the lipophilic moiety contains saturated or unsaturated C6-C6 groups. 18 Hydrocarbon chains (e.g., straight-chain C6-C) 18 (alkyl or alkenyl). In one embodiment, the lipophilic moiety contains saturated or unsaturated C. 16 Hydrocarbon chains (e.g., straight-chain C) 16 (alkyl or alkenyl).

[0131] In one embodiment of this application, the lipophilic portion is C6-C. 30 Acids (e.g., hexanoic acid, heptanoic acid, octanoic acid, nonanoic acid, decanoic acid, undecanoic acid, dodecanoic acid, tridecanoic acid, tetradecanoic acid, pentadecanoic acid, hexadecanoic acid, heptadecanoic acid, octadecanoic acid, oleic acid, linoleic acid, arachidonic acid, cis-4,7,10,13,16,19-docosahexaenoic acid, etc.) or C6-C 30 Alcohols (e.g., hexanol, heptanol, octanol, nonanol, decanol, undecanol, dodecanol, tridecanol, tetradecanol, pentadecanol, hexadecanol, heptadecanol, octadecanol, oleyl alcohol, linolenic acid, arachidonic acid, cis-4,7,10,13,16,19-docosahexaenoic acid, retinol, vitamin E, cholesterol, etc.).

[0132] In one embodiment of this application, the lipophilic portion may conjugate to the iRNA agent via direct attachment to the ribose of the iRNA agent. Alternatively, the lipophilic portion may conjugate to the iRNA agent via a linker or a vector.

[0133] In one embodiment of this application, the lipophilic portion is conjugated to the iRNA agent via one or more linkers (ligands).

[0134] In one embodiment of this application, the lipophilic portion is conjugated to a double-stranded iRNA agent via a linker containing an ether, thioether, urea, carbonate, amine, amide, maleimide-thioether, disulfide, phosphate diester, sulfonamide bond, product of click reaction (e.g., triazole from azide-alkyne cycloaddition), or carbamate.

[0135] In one embodiment of this application, at least one connector is a redox-cleavable connector (such as a reductively cleavable connector, for example, a disulfide group), an acid-cleavable connector (e.g., an hydrazone group, an ester group, an acetal group, or a ketal group), an esterase-cleavable connector (e.g., an ester group), a phosphatase-cleavable connector (e.g., a phosphate ester), or a peptidase-cleavable connector (e.g., a peptide bond).

[0136] In one embodiment of this application, at least one adapter is a biolytic adapter selected from the group consisting of: DNA, RNA, disulfides, amides, functionalized monosaccharides or oligosaccharides of galactosamine, glucosamine, glucose, galactose, and mannose, or combinations thereof.

[0137] In one embodiment of this application, the lipophilic portion is conjugated to a double-stranded iRNA agent via a carrier that replaces one or more nucleotides. The carrier can be a cyclic group or an acyclic group. In one embodiment, the cyclic group is selected from the group consisting of: pyrrolidinyl, pyrazolinyl, pyrazolinyl, imidazolinyl, imidazolinyl, piperidinyl, piperazinyl, [1,3]dioxolane, oxazolinyl, isoxazolinyl, morpholinyl, thiazolinyl, isothiazolinyl, quinoxalinyl, pyridazinoneyl, tetrahydrofuranyl, and decahydronaphthalene. In one embodiment, the acyclic group is a portion based on a serine backbone or a diethanolamine backbone.

[0138] In one embodiment of this application, the lipophilic portion is C16, where C16 represents 2'-O-hexadecyl, a short lipid chain attached to siRNA that is lipophilic and can interact with cell membranes or membrane proteins. The structural formula of the 2'-O-hexadecyl modified nucleotide is shown in formula (36).

[0139] In one embodiment of this application, C16 may be attached to any nucleotide of the positive strand.

[0140] The lipophilic portion is described in more detail in WO2019217459A1 and WO2024216155A1 (the entire contents of which are incorporated herein by reference).

[0141] In one embodiment of this application, the siRNA introduces modified nucleotides by using nucleotide monomers with corresponding modifications.

[0142] This application also provides a product for inhibiting ANGPTL7, the product comprising an active ingredient and a pharmaceutically acceptable carrier, wherein the active ingredient is the above-mentioned siRNA or the above-mentioned modified siRNA.

[0143] In one embodiment of this application, the product is a pharmaceutical composition or a kit.

[0144] In one embodiment of this application, the product is a pharmaceutical composition, and the pharmaceutically acceptable carrier can be a carrier conventionally used in the field of siRNA delivery, such as, but not limited to, magnetic nanoparticles (e.g., Fe3O4 or Fe2O3-based nanoparticles), carbon nanotubes, mesoporous silicon, calcium phosphate nanoparticles, polyethylenimine (PEI), polyamidoamine (PAMAM) dendrimer, poly(L-lysine) (PLL), chitosan, 1,2-dioleoyl-3-trimethylammonium-propane (DOTAP), poly(D&L-lactic / glycolic acid) copolymer (PLGA), and poly(2-aminoethyl ethylene phosphate). One or more of the following: phosphate), PPEEA, and poly(2-dimethylaminoethylmethacrylate), PDMAEMA, and their derivatives.

[0145] In one embodiment of this application, there are no special requirements for the content of siRNA and pharmaceutically acceptable vector; the content of each component can be the conventional content.

[0146] In one embodiment of this application, the weight ratio of the active ingredient to the pharmaceutically acceptable carrier in the pharmaceutical composition is 1:(1-500).

[0147] In one embodiment of this application, the weight ratio of the active ingredient to the pharmaceutically acceptable carrier in the pharmaceutical composition is 1:(1-50).

[0148] In one embodiment of this application, the pharmaceutical composition may further contain other pharmaceutically acceptable excipients, which may be one or more of various formulations or compounds conventionally used in the art.

[0149] In one embodiment of this application, the other pharmaceutically acceptable excipients may include at least one of pH buffers, protectants, and osmotic pressure regulators.

[0150] In one embodiment of this application, the pH buffer may be a tris(hydroxymethyl)aminomethane hydrochloride buffer with a pH of 7.5 to 8.5 and / or a phosphate buffer with a pH of 5.5 to 8.5.

[0151] In one embodiment of this application, the protective agent may be at least one selected from inositol, sorbitol, sucrose, trehalose, mannose, maltose, lactose, and glucose.

[0152] In one embodiment of this application, the content of the protective agent may be 0.01 to 30% by weight, based on the total weight of the pharmaceutical composition.

[0153] In one embodiment of this application, the osmotic pressure regulator may be sodium chloride and / or potassium chloride.

[0154] In one embodiment of this application, the content of the osmotic pressure regulator is such that the osmotic pressure of the pharmaceutical composition is 200 to 700 mOSM / L. The content of the osmotic pressure regulator can be easily determined by those skilled in the art based on the required osmotic pressure.

[0155] In one embodiment of this application, the pharmaceutical composition may be administered by any method known in the art, including but not limited to oral, intraperitoneal, or parenteral routes, including intracranial (e.g., intraventricular, intraparenchymal, and intrasheathal), intravenous, intramuscular, intravitreal, subcutaneous, transdermal, airway (aerosol), nasal, rectal, and local (including oral and sublingual) administration.

[0156] In one embodiment of this application, the pharmaceutical composition may be a liquid formulation, such as an injection; or it may be a lyophilized powder for injection, which is mixed with liquid excipients to form a liquid formulation when administered.

[0157] In one embodiment of this application, the liquid formulation may be administered, but is not limited to, subcutaneous, intramuscular, intradermal, or intravenous injection; it may also be administered, but is not limited to, via spray to the lungs, or via spray to other organs or tissues (such as the liver); it may also be administered, but is not limited to, via intradermal injection, application, or spray to the skin; or it may be, but is not limited to, via intrathecal or intravitreal injection, optionally infused into the brain (e.g., the striatum), for example, via a continuous pump infusion.

[0158] In one embodiment of this application, the pharmaceutical composition may be in the form of a liposome formulation.

[0159] In one embodiment of this application, the pharmaceutically acceptable carrier used in the liposome formulation comprises an amine-containing transfection compound (hereinafter also referred to as an organic amine), an auxiliary lipid, and / or a polyethylene glycol-modified lipid.

[0160] In one embodiment of this application, the organic amine, auxiliary lipid, and polyethylene glycol-modified lipid may be selected from one or more of the amine-containing transfection compounds or their pharmaceutically acceptable salts or derivatives, auxiliary lipids, and polyethylene glycol-modified lipids described in CN108220295B (which is incorporated herein by reference in its entirety).

[0161] In one embodiment of this application, the kit further comprises a pharmaceutically acceptable carrier and / or excipients.

[0162] In one embodiment of this application, the siRNA, pharmaceutically acceptable carrier, and / or excipients in the kit may be present individually, in a mixture of two or more of them, or in the form of a final pharmaceutical composition.

[0163] In one embodiment of this application, the pharmaceutically acceptable carrier in the kit is an amine-containing compound, an auxiliary lipid, or a polyethylene glycol-modified lipid.

[0164] In one embodiment of this application, the pharmaceutically acceptable carrier in the kit is either a mixture or exists independently.

[0165] In one embodiment of this application, the siRNA, pharmaceutically acceptable carrier, and / or excipients in the kit are provided in liquid, dry, or lyophilized form.

[0166] In one embodiment of this application, the siRNA, pharmaceutically acceptable carriers, and / or excipients in the kit are substantially pure and / or sterile.

[0167] In one embodiment of this application, the kit includes a container for providing siRNA, one or more containers for providing amine-containing compounds, auxiliary lipids, and polyethylene glycol-modified lipids, and optionally, a container for providing excipients.

[0168] In one embodiment of this application, the kit further comprises one or more components necessary or beneficial for a particular application, the components being selected from:

[0169] One or more components for achieving the desired cell transfection;

[0170] One or more components used to diagnose, treat, or prevent a specific disease or physical disorder;

[0171] One or more buffers;

[0172] Positive or negative control samples;

[0173] Excipients, stabilizers, or preservatives.

[0174] In one embodiment of this application, the one or more components for diagnosing, treating or preventing a specific disease or physical disorder are one or more additional therapeutic compounds or compositions, or one or more diagnostic reagents.

[0175] In one embodiment of this application, the kit further comprises one or more of sterile water, physiological saline, and PBS.

[0176] This application also provides the use of the above-mentioned siRNA or the above-mentioned product in the preparation of products for the prevention, diagnosis and / or treatment of pathological conditions or diseases caused by ANGPTL7.

[0177] This application also provides the use of the above-mentioned siRNA or the above-mentioned product in the preparation of products for the prevention, diagnosis and / or treatment of pathological conditions or diseases caused by ANGPTL7.

[0178] In some embodiments of this application, the ANGPTL7-related pathological conditions or diseases involve ANGPTL7 overexpression. ANGPTL7 overexpression refers to ANGPTL7 levels (e.g., ANGPTL7 levels present in the plasma or tissues of a subject, and optionally in damaged tissues) that are higher than normal ANGPTL7 levels (e.g., the corresponding levels in healthy controls).

[0179] In some implementations, the ANGPTL7-related pathological conditions or diseases will benefit from reduced ANGPTL7 levels or suppressed expression.

[0180] In some embodiments, the condition associated with ANGPTL7 expression is glaucoma or a glaucoma-related condition. Non-limiting examples of glaucoma or glaucoma-related conditions that can be treated using the methods described herein include glaucoma, open-angle glaucoma, primary open-angle glaucoma, angle-closure glaucoma, ocular inflammation, systemic inflammation, anterior uveitis, acute retinal necrosis, Sturge-Weber syndrome, Axenfeld-Rieger syndrome, Marfan syndrome, homocystinuria, Weill-Marchesani syndrome, and autoimmune diseases such as juvenile rheumatoid arthritis and Marie Strumpell's ankylosing spondylitis.

[0181] In some embodiments of this application, the ANGPTL7-related pathological conditions or diseases involve ANGPTL7 overexpression. ANGPTL7 overexpression refers to ANGPTL7 levels (e.g., ANGPTL7 levels present in the plasma or tissues of a subject, and optionally in damaged tissues) that are higher than normal ANGPTL7 levels (e.g., the corresponding levels in healthy controls).

[0182] In some implementations, the ANGPTL7-related pathological conditions or diseases will benefit from reduced ANGPTL7 levels or suppressed expression.

[0183] In some embodiments, the condition associated with ANGPTL7 expression is glaucoma or a glaucoma-related condition. Non-limiting examples of glaucoma or glaucoma-related conditions that can be treated using the methods described herein include glaucoma, open-angle glaucoma, primary open-angle glaucoma, angle-closure glaucoma, ocular inflammation, systemic inflammation, anterior uveitis, acute retinal necrosis, Sturge-Weber syndrome, Axenfeld-Rieger syndrome, Marfan syndrome, homocystinuria, Weill-Marchesani syndrome, and autoimmune diseases such as juvenile rheumatoid arthritis and Marie Strumpell's ankylosing spondylitis.

[0184] The technical solution of this application has the following advantages:

[0185] This application provides a modified siRNA for inhibiting ANGPTL7, wherein the modifications include methoxy modification, fluorination modification, thiophosphate linkage, and thermal instability modification. Activity was detected in SK-MEL-2 cells, and the siRNA showed high inhibitory activity against ANGPTL7. Detailed Implementation

[0186] The following embodiments are provided to better understand this application and are not limited to the preferred embodiments described herein. They do not constitute a limitation on the content and scope of protection of this application. Any product that is the same as or similar to this application, derived by anyone under the guidance of this application or by combining features of this application with other prior art, falls within the scope of protection of this application.

[0187] In the following embodiments, ANGPTL7 mRNA refers to mRNA having the sequences shown in GenBank accessions NM_021146.4, NM_001039554.3, NM_001103825.4, NM_001420997.1, and XM_005544804.4. Further, unless otherwise specified, the term "target gene" as used in this disclosure refers to a gene capable of transcribing the aforementioned ANGPTL7 mRNA, and the term "target mRNA" refers to the aforementioned ANGPTL7 mRNA.

[0188] For any experimental steps or conditions not specified in the following examples, the procedures or conditions described in the literature in this field can be followed. Reagents or instruments whose manufacturers are not specified are all commercially available conventional reagent products.

[0189] In the following examples, uppercase letters C, G, U, and A represent ribonucleotides; lowercase letter m indicates that the nucleotide adjacent to the left of letter m is a methoxy-modified nucleotide; lowercase letter f indicates that the nucleotide adjacent to the left of letter f is a fluorinated nucleotide; and lowercase letter s indicates that the two nucleotides adjacent to the left and right of letter s are modified with a thiophosphate group.

[0190] In the following examples, "modified nucleotide" refers to a nucleotide or nucleotide analog formed by replacing the 2'-hydroxyl group of the ribosyl group with another group, or a nucleotide whose bases are modified bases. "Fluorinated nucleotide" refers to a nucleotide formed by replacing the 2'-hydroxyl group of the ribosyl group with fluorine, and "non-fluorinated nucleotide" refers to a nucleotide or nucleotide analog formed by replacing the 2'-hydroxyl group of the ribosyl group with a non-fluorinated group. "Nucleotide analog" refers to a group that can replace a nucleotide in nucleic acids but has a structure different from adenine ribonucleotide, guanine ribonucleotide, cytosine ribonucleotide, uracil ribonucleotide, or thymine deoxyribonucleotide. Examples include isonucleotides, bridged nucleic acids (BNA), or acyclic nucleotides. "Methoxylated nucleotide" refers to a nucleotide formed by replacing the 2'-hydroxyl group of the ribosyl group with a methoxy group.

[0191] In the following examples, the terms "complementary" or "reverse complementary" are used interchangeably and have the meaning known to those skilled in the art: in a double-stranded nucleic acid molecule, the bases of one strand are paired complementaryly with the bases of the other strand. In DNA, the purine base adenine (A) always pairs with the pyrimidine base thymine (T) (or uracil (U) in RNA); the purine base guanine (C) always pairs with the pyrimidine base cytosine (G). Each base pair consists of one purine and one pyrimidine. When adenine on one strand always pairs with thymine (or uracil) on the other strand, and guanine always pairs with cytosine, the two strands are considered complementary, and the sequence of the strand can be inferred from the sequence of its complementary strand.

[0192] In the following examples, particularly in describing methods for preparing siRNA, pharmaceutical compositions, or siRNA conjugates of this disclosure, unless otherwise specified, a nucleoside monomer refers to a modified or unmodified RNA phosphoramidites (sometimes also called nucleoside phosphoramidites) used in phosphoramidite solid-phase synthesis, depending on the type and sequence of nucleotides in the siRNA or siRNA conjugate to be prepared. Phosphoramidite solid-phase synthesis is a method known to those skilled in the art for RNA synthesis. All nucleoside monomers used in this disclosure are commercially available.

[0193] In the following embodiments, "coupling" refers to the covalent connection between two or more chemical parts, each with a specific function; correspondingly, "coupling" refers to a compound formed by the covalent connection between these chemical parts. Further, "siRNA conjugate" refers to a compound formed by the covalent attachment of one or more chemical parts with specific functions to siRNA. The term "siRNA conjugate" should be understood, depending on the context, as a collective term for multiple siRNA conjugates or a siRNA conjugate represented by a specific chemical formula. In the context of this disclosure, "coupling molecule" should be understood as a specific compound that can be reactively coupled to siRNA to ultimately form the siRNA conjugate of this disclosure.

[0194] In the following examples, "optional" or "optionally" means that the event or condition described thereafter may or may not occur, and the description includes both the possibility that the event or condition occurs and the possibility that it does not occur. For example, "optionally substituted" "alkyl" includes "alkyl" and "substituted alkyl" as defined below. Those skilled in the art will understand that for any group containing one or more substituents, these groups are not intended to introduce any substitution or substitution pattern that is spatially impractical, synthetically infeasible, and / or inherently unstable.

[0195] In the following embodiments, the terms “treatment,” “relief,” or “improvement” may be used interchangeably herein. These terms refer to methods of achieving beneficial or desired outcomes, including but not limited to therapeutic benefits. A “therapeutic benefit” means the eradication or improvement of the underlying disorder being treated. Furthermore, a therapeutic benefit is achieved by eradicating or improving one or more physical symptoms associated with the underlying disorder, thereby observing improvement in the subject, although the subject may still be suffering from the underlying disorder.

[0196] In the following examples, "prevention" and "protection" are used interchangeably. These terms refer to methods of obtaining a beneficial or desired outcome, including but not limited to preventive benefits. To obtain a "preventive benefit," the composition may be given to a subject at risk of developing a specific disease, or to a subject who reports one or more pathological symptoms of a disease, even if a diagnosis of the disease may not have been made.

[0197] Unless otherwise specified, the reagents and culture media used in the following examples are all commercially available products, and the nucleic acid electrophoresis, real-time PCR and other operations used are all performed in accordance with the methods described in Molecular Biology (4th Edition) (Alexander McLennan et al., 2019).

[0198] The siRNAs involved in the following examples are siRNA sequences synthesized via phosphoramide solid-phase synthesis.

[0199] In the following examples, when transfecting cells with siRNA, siRNA conjugates, or siRNA or siRNA conjugates targeting the ANGPTL7 gene, Lipo3000 or RNAiMAX (purchased from Invitrogen) were used as transfection reagents. Specific procedures were followed according to the manufacturer's instructions. For qPCR detection, HiScript III RT SuperMix for qPCR (purchased from Vazyme) was used as the reverse transcription reagent. Specific procedures were followed according to the manufacturer's instructions.

[0200] In the following examples, uppercase letters C, G, U, and A represent ribonucleotides; lowercase letter m indicates that the nucleotide adjacent to the left of letter m is a methoxy-modified nucleotide; lowercase letter f indicates that the nucleotide adjacent to the left of letter f is a fluorinated nucleotide; lowercase letter s indicates that the two nucleotides adjacent to the left and right of letter s are modified with a thiophosphate group; string (d) indicates that the nucleotide adjacent to the left of string (d) is a 2'-deoxynucleotide modified nucleotide. "2'-deoxynucleotide modified nucleotides" include five types: A(d), C(d), G(d), U(d), and T(d). Specifically, A(d) represents 2'-deoxyadenosine-3'-phosphate, C(d) represents 2'-deoxycytidine-3'-phosphate, U(d) represents 2'-deoxyuridine-3'-phosphate, G(d) represents 2'-deoxyguanosine-3'-phosphate, and T(d) represents 2'-deoxythymidine-3'-phosphate. (When the sequence contains nucleotides modified with 2'-deoxynucleotides, if the ribonucleotide is uracil ribonucleotide, then thymine deoxyribonucleotide is used instead of uracil ribonucleotide.) The string (GNA) indicates that the nucleotide adjacent to the left of the string (GNA) is a nucleotide modified with glycol nucleic acid nucleotide. A (GNA) represents adenosine-diol nucleic acid (GNA), C (GNA) represents cytidine-diol nucleic acid (GNA), G (GNA) represents guanosine-diol nucleic acid (GNA), T (GNA) represents thymidine-diol nucleic acid (GNA), and U (GNA) represents uridine-diol nucleic acid (GNA).

[0201] Unless otherwise specified, all reagent ratios provided below are calculated on a volume ratio (v / v).

[0202] Example 1: A siRNA for inhibiting ANGPTL7

[0203] This embodiment provides an siRNA for inhibiting ANGPTL7. The nucleotide sequence of the siRNA is designed based on the target mRNA, as shown in Table 1. The siRNA molecule with the following sequence was synthesized by Suzhou Genegene Co., Ltd.

[0204] Table 1. Nucleotide sequences of siRNAs that inhibit ANGPTL7

[0205] Example 2: A modified siRNA for inhibiting ANGPTL7

[0206] This embodiment provides a modified siRNA for inhibiting ANGPTL7. The siRNA is modified based on Example 1, and the modified sequence is shown in Table 2.

[0207] Taking hANGPTL7_1L23M2 as an example, it is a modification based on the sequence hANGPTL7_1. According to Table 2, following the direction from the 5' end to the 3' end, the nucleotides at positions 7, 9, 10, and 11 of the positive strand are fluorinated nucleotides, and the nucleotides at positions 1, 2, 3, 4, 5, 6, 8, 12, 13, 14, 15, 16, 17, 18, 19, 20, and 21 of the positive strand are methoxylated nucleotides. Positions 1 and 2, and 2 and 3 are linked by thiophosphate groups.

[0208] Following the direction from the 5' end to the 3' end, the nucleotides at positions 2, 6, 14, and 16 of the antisense strand are fluorinated nucleotides, and the nucleotides at positions 1, 3, 4, 5, 7, 8, 9, 10, 11, 12, 13, 15, 17, 18, 19, 20, 21, 22, and 23 of the antisense strand are methoxylated nucleotides; the nucleotides at positions 1 and 2, 2 and 3, the penultimate and penultimate positions, and the penultimate and penultimate positions are linked by thiophosphate groups.

[0209] Table 2. Nucleotide sequences of modified siRNAs that inhibit ANGPTL7

[0210] Experimental Example 1: Activity assay of modified siRNA for inhibiting ANGPTL7 in SK-MEL-2 cells

[0211] This experimental example provides the determination of the relative inhibitory level of the siRNA compound of this application on ANGPTL7 mRNA in SK-MEL-2 cells by quantitative real-time PCR (qPCR).

[0212] The experiment used to detect the activity of ANGPTL7-modified siRNA in SK-MEL-2 cells was conducted as follows:

[0213] 1. SK-MEL-2 cells (human skin melanoma cells, purchased from the Cell Bank of the Chinese Academy of Sciences) were seeded into DMEM medium (GIBCO catalog number: 10566016) containing 10% (v / v) fetal bovine serum (FBS, purchased from Hyclone) and cultured in a 5% (v / v) CO2 incubator at 37°C for 48 h. After culture, the SK-MEL-2 cells were digested with trypsin (purchased from GIBCO, catalog number 25200-072). After digestion, the cells were first rinsed with PBS buffer and then resuspended in DMEM medium to obtain a cell concentration of 3 × 10⁻⁶ cells / year. 5 Cell suspension of cells / mL;

[0214] 2. Dilute the different siRNAs listed in Table 3 using opti-MEM (Gibco, catalog number 31985-070) to obtain siRNA dilution solutions containing different siRNAs; mix 25 μL opti-MEM with 0.25 μL Lipofectamine RNAiMAX transfection reagent (Thermo Fisher Scientific, catalog number 11668-019) to obtain transfection reagent dilution solution; take 25 μL of siRNA dilution solution containing different siRNAs and mix it with the transfection reagent dilution solution, and let it stand at room temperature (25℃) for 15 min to obtain transfection solutions containing different siRNAs;

[0215] 3. After seeding the cell suspension into 96-well plates at a seeding rate of 50 μL / well, set up MOCK group (MOCK group is the transfection reagent control group), hANGPTL7_1L23M2 group to hANGPTL7_255L23M2 group in 96-well plates, with 3 replicates in each group.

[0216] 4. After setting up, add 50 μL of transfection buffer containing different siRNAs to the wells of each experimental group (the hhANGPTL7_1L23M2 experimental group was given transfection buffer containing hANGPTL7_1L23M2, the hANGPTL7_2L23M2 experimental group was given transfection buffer containing hANGPTL7_2L23M2, and so on, with the final concentration of siRNA in the wells being 10 nM). Incubate the cells in a 5% (v / v) CO2, 37℃ cell culture incubator for 24 h for transfection. After transfection, discard the liquid in the wells and collect the cells.

[0217] 5. Total RNA was extracted from cells in each well using a magnetic bead method total RNA extraction kit (purchased from Suzhou Genecast Co., Ltd., catalog number E31008) according to the method described in the kit instructions.

[0218] 6. RNA reverse transcription

[0219] The experimental procedure was performed using HiScript III RT SuperMix for qPCR (purchased from Novizan, catalog number R323-01) following the product instructions. A 20 μL reverse transcription reaction system was prepared according to the reverse transcription procedure in the kit instructions to reverse transcribe total RNA from cells. The reverse transcription conditions were as follows: the reverse transcription reaction system was incubated at 37°C for 15 min, then at 85°C for 5 s. 80 μL of DEPC water was added to each reverse transcription reaction system to obtain a solution containing cDNA.

[0220] 7. Preparation of qPCR reaction system

[0221] For each reverse transcription reaction system, take 4 μL of the above-mentioned cDNA-containing solution as a template. Using the reagents provided in the AceQ Universal SYBR qPCR Master Mix kit (purchased from Vazyme, catalog number Q511-02), prepare a 20 μL qPCR reaction system on an ice box according to Table 4. Primer 1 and Primer 2 are the PCR primer sequences for amplifying the target gene ANGPTL7 and the internal reference gene GAPDH, respectively (as shown in Table 5). Place each qPCR reaction system in an ABIStepOnePlus Real-Time... On a PCR instrument, a three-step amplification method was used. The amplification program was 95℃ pre-denaturation for 10 min, followed by 95℃ denaturation for 30 s, 60℃ annealing for 30 s, and 72℃ extension for 30 s. This denaturation, annealing, and extension process was repeated 40 times to obtain product W containing the amplified target gene ANGPTL7 and the internal reference gene GAPDH. Product W was then incubated sequentially at 95℃ for 15 s, 60℃ for 1 min, and 95℃ for 15 s. The melting curves of the target gene ANGPTL7 and the internal reference gene GAPDH in product W were collected by a real-time quantitative PCR instrument to obtain the Ct values ​​of the target gene ANGPTL7 and the internal reference gene GAPDH.

[0222] Table 4. DNA Amplification Reaction System

[0223] Table 5. Primer Information

[0224] The relative quantification of the target gene ANGPTL7 in each test group was performed using the Ct(ΔΔCt) method, as follows:

[0225] ΔCt(test group) = Ct(target gene in test group) – Ct(internal reference gene in test group)

[0226] ΔCt(control group) = Ct(target gene in control group) – Ct(internal reference gene in control group)

[0227] ΔCt(test group) = ΔCt(test group) - ΔCt(control group average)

[0228] ΔCt(control group) = ΔCt(control group) - ΔCt(control group average)

[0229] Wherein, ΔCt (control group mean) is the arithmetic mean of ΔCt (control group) for each sample in the control group; thus, each sample in the test group and the control group corresponds to a ΔCt value.

[0230] Using the control group as a baseline, the expression level of ANGPTL7 mRNA in the test group was normalized, and the expression level of ANGPTL7 mRNA in the control group was defined as 100%.

[0231] The relative expression level of ANGPTL7 mRNA in the test group = 2 - ΔΔCt(test group) × 100%

[0232] The inhibition rate of ANGPTL7 mRNA in the test group = 1 - the relative expression level of ANGPTL7 mRNA in the test group.

[0233] The ANGPTL7 mRNA level was compared with the internal reference gene GAPDH, and the value was normalized to the mean of the saline control group. The data were expressed as a percentage relative to the saline control group and presented as the mean plus the standard deviation.

[0234] The experimental results are shown in Table 6.

[0235] Table 6. Inhibition level of siRNA at 10 nM mRNA in SK-MEL-2 cells.

[0236] Obviously, the above embodiments are merely illustrative examples for clear explanation and are not intended to limit the implementation. Those skilled in the art will recognize that other variations or modifications can be made based on the above description. It is neither necessary nor possible to exhaustively list all possible implementations here. However, obvious variations or modifications derived therefrom are still within the scope of protection of this invention.

Claims

1. A siRNA for inhibiting ANGPTL7, characterized in that, The siRNA contains a sense strand and an antisense strand, which are at least partially anticomplementary to form a double-stranded region; the sense strand of the siRNA comprises at least 15 consecutive nucleotides with nucleotide sequences as shown in any one of the nucleic acid molecules or nucleic acid sequences shown in any one of SEQ ID NO. 1 to 255, differing by no more than 3 nucleotides; the antisense strand of the siRNA comprises at least 15 consecutive nucleotides with nucleotide sequences as shown in any one of the nucleic acid molecules or nucleic acid sequences shown in any one of SEQ ID NO. 256 to 510, differing by no more than 3 nucleotides.

2. The siRNA as described in claim 1, characterized in that, At least one nucleotide in the sense or antisense strand of the siRNA is a modified nucleotide.

3. The siRNA as described in claim 2, characterized in that, The modified nucleotide is a compound formed by modifying the ribose of a nucleotide, or a compound formed by modifying the backbone between nucleotides, or a compound formed by modifying the bases on a nucleotide, or a compound formed by modifying the ends of siRNA.

4. The siRNA as described in claim 3, characterized in that, The modified nucleotides include methoxylated nucleotides, fluorinated nucleotides, phosphate-thioester linked nucleotides, deoxyribonucleotide-substituted nucleotides, 2'-deoxy-nucleotides, 3'-terminal deoxy-thymidine nucleotides, 3'-deoxy-nucleotides, inosine nucleotides, locked nucleotides, non-locked nucleotides, configuration-restricted nucleotides, restricted ethyl nucleotides, baseless nucleotides, reverse nucleotides, reverse baseless nucleotides, vinyl phosphate-modified nucleotides, 2'-amino-modified nucleotides, 2'-O-allyl-modified nucleotides, 2'-C-alkyl-modified nucleotides, and 2'-hydroxy-modified nucleotides. - Modified nucleotides, 2'-methoxyethyl modified nucleotides, 2'-O-alkyl- modified nucleotides, morpholinonucleotide modified nucleotides, aminophosphate modified nucleotides, nucleotides containing non-natural bases, tetrahydropyran modified nucleotides, 1,5-dehydrohexyl alcohol modified nucleotides, cyclohexenyl modified nucleotides, ethylene glycol nucleic acid modified nucleotides, open-ring nucleotide modified nucleotides, nucleotides containing methyl phosphate groups, nucleotides containing 5'-phosphate groups, nucleotides containing 5'-phosphate mimics, terminal nucleotides linked to cholesterol derivatives, and terminal nucleotides linked to dodecanoic acid bisdecamide groups.

5. The siRNA according to any one of claims 2 to 4, characterized in that, Fluorinated nucleotides are located in both the antisense and sense strands of the nucleotide sequence. Specifically, in the sense strand, positions 7, 8, and 9, or positions 5, 7, 8, and 9, or positions 9, 10, and 11, or positions 7, 9, 10, and 11, are fluorinated nucleotides, while the remaining nucleotide groups in the sense strand are non-fluorinated nucleotides. In the antisense strand, positions 2, 6, 12, 14, and 16, or positions 2, 8, 12, 14, and 16, or positions 2, 9, 12, 14, and 16, or positions 2, 10, 12, and 14, are fluorinated nucleotides. The nucleotides at positions 16, 2, 6, 10, 12, 14, 2, 8, 10, 14, 16, 2, 6, 10, 14, 16, 2, 4, 12, 14, 16, 2, 6, 14, 16, 2, 10, 12, 14, 16, 18, 20, 2, 6, 8, 14, 16, 14, 10, 12, 14, 16, 18, 20, 2, 14, 16 are fluorinated nucleotides, and the nucleotide groups at the remaining positions in the antisense strand are each non-fluorinated nucleotide groups. Each of the non-fluorinated modified nucleotide groups is independently selected from one of the nucleotide groups or nucleotide analog groups formed by replacing the 2' hydroxyl group of the ribosyl group with a non-fluorinated group, wherein the non-fluorinated modified nucleotide is a methoxy-modified nucleotide and / or a 2′-deoxynucleotide and / or an inosine nucleotide and / or a ethylene glycol-modified nucleotide.

6. The siRNA according to any one of claims 2 to 5, characterized in that, In the direction from the 5' end to the 3' end, at least one nucleotide in the nucleotides at positions 2 to 18 of the antisense strand is a heat-labile modified nucleotide; optionally, in the direction from the 5' end to the 3' end, at least one nucleotide in the nucleotides at positions 5, 7, 10, 11, 12, 13, and 16 of the antisense strand is a heat-labile modified nucleotide.

7. The siRNA according to any one of claims 2 to 6, characterized in that, In the direction from the 5' end to the 3' end, at least two nucleotides in the antisense strand 2 to 8 are thermally unstable modified nucleotides.

8. The siRNA according to any one of claims 2 to 7, characterized in that, The heat-labile modified nucleotides are ethylene glycol-modified nucleotides (GNA) and / or 2′-deoxynucleotides.

9. The siRNA according to any one of claims 2 to 8, characterized in that, The non-fluorinated modified nucleotide group is a methoxylated modified nucleotide.

10. The siRNA according to any one of claims 2 to 9, characterized in that, The nucleotides linked by the thiophosphate group are located in the antisense and sense strands of the nucleotide sequence, and, in the direction from the 5' end to the 3' end, at least the nucleotides at the 1st and 2nd, 2nd and 3rd, penultimate and penultimate, and penultimate and penultimate positions of the antisense strand are linked by the thiophosphate group. Alternatively, in the direction from the 5' end to the 3' end, at least the first and second nucleotides of the positive strand are linked by a phosphate thioester group; Alternatively, in the direction from the 5' end to the 3' end, at least the nucleotides at positions 1 and 2, and positions 2 and 3 of the sense strand are linked by thiophosphate groups, and at least the nucleotides at positions 1 and 2, positions 2 and 3, positions 1 to 1 and 2 to 1, and positions 2 to 1 to 3 to 1 of the antisense strand are linked by thiophosphate groups. Alternatively, in the direction from the 5' end to the 3' end, at least the first and second positions of the sense strand are linked by a thiophosphate group, and at least the first and second, second and third, penultimate and penultimate, and penultimate and penultimate nucleotides of the antisense strand are linked by a thiophosphate group. Alternatively, in the direction from the 5' end to the 3' end, at least the first and second positions, and the 20th and 21st positions of the sense strand are linked by thiophosphate groups, and at least the first and second positions, the second and third positions, the penultimate and penultimate positions, and the penultimate and penultimate positions of the antisense strand are linked by thiophosphate groups. Alternatively, in the direction from the 5' end to the 3' end, at least the nucleotides at positions 1 and 2, 2 and 3, penultimate and penultimate, and penultimate and penultimate of the sense strand are linked by thiophosphate groups, and at least the nucleotides at positions 1 and 2, 2 and 3, penultimate and penultimate, and penultimate and penultimate of the antisense strand are linked by thiophosphate groups. Alternatively, in the direction from the 5' end to the 3' end, the nucleotides linked by the thiophosphate group are located in the antisense and sense strands of the nucleotide sequence, and in the direction from the 5' end to the 3' end, at least the nucleotides at positions 1 and 2, 19 and 20, and 20 and 21 of the sense strand are linked by thiophosphate groups, and at least the nucleotides at positions 1 and 2, 2 and 3, penultimate and penultimate, and penultimate and penultimate of the antisense strand are linked by thiophosphate groups.

11. The siRNA according to any one of claims 2 to 10, characterized in that, The 5' and / or 3' ends of the siRNA's sense and / or antisense strands contain at least one reverse-abase-free nucleotide.

12. The siRNA according to any one of claims 2 to 11, characterized in that, The first nucleotide at the 5' and / or 3' end of the siRNA's sense and / or antisense strands is a vinyl phosphate-modified nucleotide.

13. The siRNA according to any one of claims 2 to 12, characterized in that, The siRNA is coupled to or conjugated with one or more nucleotides on at least one strand via a linker or vector, or with one or more lipophilic moieties.

14. The siRNA according to any one of claims 2 to 12, characterized in that, The siRNA has a ligand coupled to the ends of its sense and / or antisense strands, and the ligand is GalNAc.

15. A product for inhibiting ANGPTL7, characterized in that, The product comprises an active ingredient and a pharmaceutically acceptable carrier, wherein the active ingredient is siRNA as described in any one of claims 1 to 14.

16. The use of the siRNA of any one of claims 1 to 14 or the product of claim 15 in the preparation of products for the prevention, diagnosis and / or treatment of pathological conditions or diseases caused by ANGPTL7.

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