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KR-20260062944-A - RNAi agents targeting APP and their medicinal uses

KR20260062944AKR 20260062944 AKR20260062944 AKR 20260062944AKR-20260062944-A

Abstract

The invention relates to RNAi agents targeting APP and their medicinal uses, specifically, to RNAi agents targeting APP, pharmaceutical compositions, and their medicinal uses. Additionally, the invention relates to pharmaceutical compositions, cells, or kits comprising said RNAi agents, and methods of using said RNAi agents to treat and/or prevent a subject suffering from a related disorder.

Inventors

  • 리우 민
  • 천 디
  • 린 샤오얀
  • 리 윈페이
  • 왕 얀후이

Assignees

  • 투오지에 바이오텍 (상하이) 컴퍼니 리미티드

Dates

Publication Date
20260507
Application Date
20240830
Priority Date
20230831

Claims (17)

  1. As an RNAi preparation, it comprises a sense strand and an antisense strand forming a double-stranded region; The sense strand comprises at least 15 consecutive nucleotide sequences that differ from any nucleotide sequence among SEQ ID NO: 1 to SEQ ID NO: 11 by three or fewer nucleotides; and An RNAi preparation comprising an antisense strand comprising at least 15 consecutive nucleotide sequences that differ from any nucleotide sequence among SEQ ID NO: 12 to SEQ ID NO: 22 by 3 or fewer nucleotides.
  2. In claim 1, the sense strand comprises at least 17 consecutive nucleotide sequences that differ by three or fewer nucleotides from any nucleotide sequence among SEQ ID NO: 1 to SEQ ID NO: 11; and the antisense strand comprises at least 17 consecutive nucleotide sequences that differ by three or fewer nucleotides from any nucleotide sequence among SEQ ID NO: 12 to SEQ ID NO: 22; Preferably, the sense strand comprises at least 19 consecutive nucleotide sequences that differ from any nucleotide sequence among SEQ ID NO: 1 to SEQ ID NO: 11 by three or fewer nucleotides; preferably, the nucleotide sequences differ by one or fewer nucleotides, and/or, The above antisense strand comprises at least 21 consecutive nucleotide sequences that differ from any nucleotide sequence among SEQ ID NO: 12 to SEQ ID NO: 22 by three or fewer nucleotides; preferably, the nucleotide sequences differ by one or fewer nucleotides, an RNAi preparation.
  3. In claim 1 or 2, the RNAi preparation comprises: hereinafter: Group 1), a sense strand as presented in SEQ ID NO: 1 and an antisense strand as presented in SEQ ID NO: 12; or a sense strand as presented in SEQ ID NO: 2 and an antisense strand as presented in SEQ ID NO: 13; Group 2), sense strand as presented in SEQ ID NO: 3 and antisense strand as presented in SEQ ID NO: 14; or a sense strand as presented in SEQ ID NO: 4 and an antisense strand as presented in SEQ ID NO: 15; Group 3), sense strand as presented in SEQ ID NO: 5 and antisense strand as presented in SEQ ID NO: 16; Group 4), sense strand as presented in SEQ ID NO: 6 and antisense strand as presented in SEQ ID NO: 17; or a sense strand as presented in SEQ ID NO: 7 and an antisense strand as presented in SEQ ID NO: 18; Group 6), sense strand as presented in SEQ ID NO: 8 and antisense strand as presented in SEQ ID NO: 19; or a sense strand as presented in SEQ ID NO: 9 and an antisense strand as presented in SEQ ID NO: 20; and Group 7), sense strand as presented in SEQ ID NO: 10 and antisense strand as presented in SEQ ID NO: 21; or a RNAi preparation comprising or composed of a combination of any of the sense strand and the antisense strand as presented in SEQ ID NO: 11 and the antisense strand as presented in SEQ ID NO: 22.
  4. In any one of claims 1 to 3, at least one nucleotide among the sense strand and/or antisense strand is a modified nucleotide; Preferably, an RNAi preparation in which all nucleotides in the sense strand and/or antisense strand are all modified nucleotides.
  5. In paragraph 4, three consecutive nucleotides in the sense strand are 2'-fluoromodified nucleotides; and/or Along the direction from the 5' end to the 3' end, the nucleotides at the 2nd, 6th, 12th, 14th, and 16th positions of the antisense strand are each independently 2'-fluoromodified nucleotides; Preferably, Along the direction from the 5' end to the 3' end, the nucleotides at the 7th, 8th, and 9th positions of the sense strand are 2'-fluoromodified nucleotides, and the nucleotides at the 2nd, 6th, 12th, 14th, and 16th positions of the antisense strand are each independently 2'-fluoromodified nucleotides; or Along the direction from the 5' end to the 3' end, the nucleotides at the 7th, 8th, and 9th positions of the sense strand are 2'-fluoromodified nucleotides, and the nucleotides at the 2nd, 4th, 6th, 10th, 12th, 14th, 16th, or 18th positions of the antisense strand are each independently 2'-fluoromodified nucleotides; and RNAi preparations in which the nucleotides at the remaining positions of the sense strand and the antisense strand are 2-methoxy modified nucleotides.
  6. In any one of claims 1 to 5, at least one of the sense strand and/or antisense strand is a phosphodiester having a modification group; Preferably, the phosphodiester having the above modification group is a phosphorothioate diester, an RNAi preparation.
  7. In paragraph 6, the phosphodiester having the above-mentioned modification group is as follows: Between the 1st nucleotide and the 2nd nucleotide at the 5' end of the sense strand; Between the 2nd and 3rd nucleotides of the 5' end of the sense strand; Between the 1st nucleotide and the 2nd nucleotide at the 3' end of the sense strand; Between the 2nd and 3rd nucleotides of the 3' end of the sense strand; Between the 1st nucleotide and the 2nd nucleotide at the 5' end of the antisense strand; Between the 2nd and 3rd nucleotides of the 5' end of the antisense strand; Between the 1st nucleotide and the 2nd nucleotide of the 3' end of the antisense strand; and RNAi preparations present at least one position selected from between the second nucleotide and the third nucleotide of the 3' end of the antisense strand.
  8. In any one of claims 1 to 7, the 5' end of the antisense strand comprises a phosphorus-containing group, and the phosphorus-containing group is a 5'-vinylphosphodiester, RNAi preparation.
  9. In any one of claims 1 to 8, the RNAi preparation is connected to one or more lipophilic groups; Preferably, the lipophilic group is connected to the 1st or 6th nucleotide of the 5' end of the sense strand, an RNAi preparation.
  10. In any one of claims 1 to 9, the sense strand comprises a nucleotide sequence presented in any one of SEQ ID NO: 109 to SEQ ID NO: 125; and/or, the antisense strand comprises a nucleotide sequence presented in any one of SEQ ID NO: 126 to SEQ ID NO: 142; Preferably, the RNAi preparation is as follows: A sense strand as presented in SEQ ID NO: 109 and an antisense strand as presented in SEQ ID NO: 126; A sense strand as presented in SEQ ID NO: 110 and an antisense strand as presented in SEQ ID NO: 127; A sense strand as presented in SEQ ID NO: 111 and an antisense strand as presented in SEQ ID NO: 128; A sense strand as presented in SEQ ID NO: 112 and an antisense strand as presented in SEQ ID NO: 129; A sense strand as presented in SEQ ID NO: 113 and an antisense strand as presented in SEQ ID NO: 130; A sense strand as presented in SEQ ID NO: 114 and an antisense strand as presented in SEQ ID NO: 131; A sense strand as presented in SEQ ID NO: 115 and an antisense strand as presented in SEQ ID NO: 132; A sense strand as presented in SEQ ID NO: 116 and an antisense strand as presented in SEQ ID NO: 133; A sense strand as presented in SEQ ID NO: 117 and an antisense strand as presented in SEQ ID NO: 134; A sense strand as presented in SEQ ID NO: 118 and an antisense strand as presented in SEQ ID NO: 135; A sense strand as presented in SEQ ID NO: 119 and an antisense strand as presented in SEQ ID NO: 136; A sense strand as presented in SEQ ID NO: 120 and an antisense strand as presented in SEQ ID NO: 137; A sense strand as presented in SEQ ID NO: 121 and an antisense strand as presented in SEQ ID NO: 138; A sense strand as presented in SEQ ID NO: 122 and an antisense strand as presented in SEQ ID NO: 139; A sense strand as presented in SEQ ID NO: 123 and an antisense strand as presented in SEQ ID NO: 140; A sense strand as presented in SEQ ID NO: 124 and an antisense strand as presented in SEQ ID NO: 141; and RNAi preparations comprising or composed of any combination of a sense strand and an antisense strand, among the sense strand as presented in SEQ ID NO: 125 and the antisense strand as presented in SEQ ID NO: 142.
  11. A pharmaceutical composition comprising an RNAi preparation according to any one of claims 1 to 10, and a pharmaceutically acceptable carrier.
  12. A cell comprising an RNAi preparation according to any one of claims 1 to 10.
  13. A kit comprising an RNAi preparation according to any one of claims 1 to 10 or a pharmaceutical composition according to claim 11.
  14. A method for reducing the expression of amyloid precursor protein (APP), comprising the step of administering to a subject an RNAi preparation according to any one of claims 1 to 10 or a pharmaceutical composition according to claim 11.
  15. A method for treating and/or preventing a disease, comprising the step of administering an RNAi preparation according to any one of claims 1 to 10 or a pharmaceutical composition according to claim 11 to a subject; preferably, said disease is a neurodegenerative disease associated with the APP gene; more preferably, said disease is selected from Alzheimer's disease, frontotemporal dementia, and cerebral amyloid angiopathy.
  16. A method for delivering an RNAi agent that inhibits APP expression and/or replication within the body, comprising the step of administering to a subject an RNAi agent according to any one of claims 1 to 10 or a pharmaceutical composition according to claim 11.
  17. A method for preparing an RNAi preparation or a pharmaceutical composition, comprising the step of synthesizing an RNAi preparation according to any one of claims 1 to 10 or a pharmaceutical composition according to claim 11.

Description

RNAi agents targeting APP and their medicinal uses The present disclosure belongs to the field of biomedical science and, specifically, relates to RNAi preparations of amyloid precursor proteins (APP), pharmaceutical compositions containing the same, and medicinal uses thereof. The amyloid precursor protein (APP) gene encodes a membrane protein expressed in neurons and glial cells. APP is cleaved by α-secreting enzymes to produce soluble and non-neurotoxic sAPPα; APP is cleaved by β- and γ-secreting enzymes to produce hydrophobic and easily aggregated Aβ polypeptides, which are secreted extracellularly. Aβ40 and Aβ42 are two common subtypes and are considered to be associated with the development and progression of Alzheimer's disease (AD) as major components of amyloid plaques. Furthermore, both Alzheimer's disease and cerebral amyloid angiopathy in patients with Down syndrome are characterized by the abnormal accumulation of amyloid plaques. By using drugs capable of selectively and effectively inhibiting APP to suppress the expression and/or activity of APP, the production and/or levels of Aβ are blocked or inhibited, which helps prevent or treat various APP-related diseases. These drugs, methods, and pharmaceutical compositions may be used to improve at least one symptom or sign of a neurodegenerative disease, including cognitive impairments such as memory and language decline; behavioral and psychological symptoms such as apathy and lack of motivation, gait disorders, and epileptic seizures; and progressive dementia and abnormal amyloid deposition. Neurodegenerative diseases are selected from sporadic Alzheimer's disease, hereditary/familial Alzheimer's disease, Down syndrome, frontotemporal dementia, and cerebral amyloid angiopathy. Currently, there is a lack of acceptable treatments for this type of neurodegenerative disease. Therefore, to effectively treat subjects suffering from APP-related diseases, drugs that selectively and effectively suppress or regulate APP gene expression are required. The purpose of this invention is to provide a method for treating this type of disease. The present disclosure provides an RNAi formulation targeting APP. In some embodiments, the present disclosure provides an RNAi formulation comprising a sense strand and an antisense strand forming a double-stranded region; The sense strand comprises a sequence that differs from any nucleotide sequence among SEQ ID NO: 1 to SEQ ID NO: 11, SEQ ID NO: 23 to SEQ ID NO: 65 by three or fewer nucleotides (e.g., 0, 1, 2, 3), and comprises at least 15 consecutive nucleotides (e.g., 16, 17, 18, 19, 20, 21); The above antisense strand comprises a sequence that differs from any nucleotide sequence among SEQ ID NO: 11 or SEQ ID NO: 22, SEQ ID NO: 66 to SEQ ID NO: 108 by three or fewer nucleotides (e.g., 0, 1, 2, 3) and comprises at least 15 consecutive nucleotides (e.g., 16, 17, 18, 19, 20, 21). In some embodiments, the antisense strand is at least partially inversely complementary to the target sequence to mediate RNA interference. In some embodiments, there are five or fewer, four or fewer, three or fewer, two or fewer, or one or fewer mismatches between the antisense strand and the target sequence. In some embodiments, the antisense strand is completely inversely complementary to the target sequence. In some embodiments, the sense strand is at least partially inversely complementary to the antisense strand to form a double-stranded region. In some embodiments, there are five or fewer, four or fewer, three or fewer, two or fewer, and one or fewer mismatches between the sense strand and the antisense strand. In some embodiments, the sense strand is completely inversely complementary to the antisense strand. In some embodiments, the RNAi formulation of the present disclosure comprises one or two smoothing units. In some specific embodiments, each strand of the RNAi preparation comprises an overhanging unit having one or two unpaired nucleotides independently. In some embodiments, the RNAi formulation of the present disclosure comprises an overhanging end located at the 3' end of the antisense strand of the RNAi formulation. In some embodiments, the sense strand and the antisense strand each independently have 16 to 35, 16 to 34, 17 to 34, 17 to 33, 18 to 33, 18 to 32, 18 to 31, 18 to 30, 18 to 29, 18 to 28, 18 to 27, 18 to 26, 18 to 25, 18 to 24, 18 to 23, 19 to 25, 19 to 24, or 19 to 23 nucleotides (e.g., 19, 20, 21, 22, 23 nucleotides). In some embodiments, the sense strand and the antisense strand have the same or different lengths, the length of the sense strand is 19 to 23 nucleotides, and the length of the antisense strand is 19 to 26 nucleotides. The length ratio of the sense strand to the antisense strand of the RNAi preparation provided in the present disclosure is 19/19, 19/20, 19/21, 19/22, 19/23, 19/24, 19/25, 19/26, 20/19, 20/20, 20/21, 20/22, 20/23, 20/24, 20/25, 20/26, 21/20, 21/21, 21/22, 21/23, 21/24, 21/25, 21/26, 22/