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EP-4741500-A1 - POLYNUCLEOTIDE, PHARMACEUTICAL COMPOSITION, AND USE THEREOF

EP4741500A1EP 4741500 A1EP4741500 A1EP 4741500A1EP-4741500-A1

Abstract

The present application discloses a polynucleotide, a pharmaceutical composition and uses thereof. In the present application, various novel polynucleotide sequences targeting the 3' untranslated region of lamin A mutant mRNA are designed, which can reduce the expression level of lamin A mutants (e.g., progerin) at the Lamin A mRNA stage, while not affecting the normal transcription of Lamin C, thereby resulting in reduced toxicity. These sequences show broad prospects in the prevention or treatment of diseases associated with lamin A mutations, particularly progeria.

Inventors

  • SHEN, Ning
  • LIANG, Jinzhao
  • LV, Lin

Assignees

  • Liangzhu Laboratory

Dates

Publication Date
20260513
Application Date
20240607

Claims (20)

  1. A polynucleotide, characterized in that the length of said polynucleotide is not less than 10 bp, and said polynucleotide comprises a polynucleotide fragment complementary to the following sequence or a fragment thereof: (i) the sequence as shown in SEQ ID NO: 11; and/or (ii) a polynucleotide sequence formed by substitution, deletion, or addition of one or several nucleotides to the sequence shown in SEQ ID NO: 11.
  2. The polynucleotide according to claim 1, characterized in that said polynucleotide comprises a polynucleotide fragment complementary to the following sequence or a fragment thereof: (i) the sequence as shown in SEQ ID NO: 1; and/or (ii) a polynucleotide sequence formed by substitution, deletion, or addition of one or several nucleotides to the sequence shown in SEQ ID NO: 1.
  3. The polynucleotide according to claim 1, characterized in that said polynucleotide comprises a polynucleotide fragment complementary to the following sequence or a fragment thereof: (i) the sequence as shown in SEQ ID NO: 10; and/or (ii) a polynucleotide sequence formed by substitution, deletion, or addition of one or several nucleotides to the sequence shown in SEQ ID NO: 10.
  4. The polynucleotide according to claim 1, characterized in that said polynucleotide comprises at least one polynucleotide fragment selected from the group consisting of: (i) the polynucleotide fragments as shown in SEQ ID NOs: 2-9; and (ii) polynucleotide fragments having at least 80% sequence identity with SEQ ID NOs: 2-9.
  5. The polynucleotide according to claim 1, characterized in that said polynucleotide is at least one selected from the group consisting of: (i) the polynucleotides as shown in SEQ ID NOs: 2-9; and (ii) polynucleotides having at least 80% sequence identity with SEQ ID NOs: 2-9.
  6. The polynucleotide according to claim 5, characterized in that said polynucleotide is the polynucleotide as shown in SEQ ID NO: 5 or the polynucleotide as shown in SEQ ID NO: 7.
  7. The polynucleotide according to any one of claims 1-6, characterized in that said polynucleotide is obtained by isolation.
  8. A pharmaceutical composition, characterized in that said pharmaceutical composition comprises the polynucleotide according to any one of claims 1 to 6, and a pharmaceutically acceptable carrier.
  9. Use of polynucleotide according to any one of claims 1 to 6 or the pharmaceutical composition according to claim 8, characterized in that said use comprises at least one of the following: (i) preventing and/or treating a disease associated with a Lamin A mutation; (ii) preparing a medicament for preventing and/or treating a disease associated with a Lamin A mutation; (iii) reducing the expression level of a Lamin A mutant; (iv) reducing the number of senescence-positive cells; (v) reducing the number of cells with dysmorphic nuclei.
  10. The use according to claim 9, characterized in that said Lamin A mutation is a mutation in a region of the LMNA gene corresponding to exon 11 and/or exon 12.
  11. The use according to claim 9, characterized in that said disease associated with a Lamin A mutation is a disease caused by a mutation occurring in exon 11 and/or exon 12 of Lamin A mRNA.
  12. The use according to claim 11, characterized in that said disease associated with a lamin A mutation is a disease caused by a mutation in lamin A mRNA at one or more positions selected from: c.1711; c.1713; c.1714; c.1718; c.1733; c.1744; c.1745; c.1748; c.1751; c.1756; c.1762; c.1772; c.1774; c.1786; c.1804; c.1821; c.1822; c.1824; c.1851; c.1868; c.1871; c.1892; c.1904; c.1916; c.1928; c.1930; c.1931; c.1940; c.1960; c.1961; c.1968; c.1968+1; c.1968+2; c.1968+5; and c.1975.
  13. The use according to claim 12, characterized in that said disease associated with a lamin A mutation is a disease caused by one or more mutations in Lamin A mRNA selected from: c.1711A>T; c.1713C>A; c.1714A>T; c.1714insCTGC; c.1718C>T; c.1733A>T; c.1744C>T; c.1745G>A; c.1748C>T; c.1751G>A; c.1756G>A; c.1762T>C; c.1772G>T; c.1774G>A; c.1786G>A; c.1804G>A; c.1821G>A; c.1822G>A; c.1824C>T; c.1851C>T; c.1868C>G; c.1871G>A; c.1892G>A; c.1904G>A; c.1916A>G; c.1928C>A; c.1930C>T; c.1931G>A; c.1940T>G; c.1960C>T; c.1961dup; c.1968G>A; c.1968+1G>A; c.1968+2T>C; c.1968+5G>A; c.1968+5G>C; and c.1975dup.
  14. The use according to claim 9, characterized in that said disease associated with a lamin A mutation is at least one selected from the group consisting of: dilated cardiomyopathy and conduction disorders (DCM-CD), muscular dystrophy, Charcot-Marie-Tooth disease type 2 (CMT2), familial partial lipodystrophy (FPLD), striated muscle laminopathy, progeroid syndrome, Emery-Dreifuss muscular dystrophy (EDMD), insulin resistance syndrome (IRS), progeria, limb-girdle muscular dystrophy (LGMD), congenital muscular dystrophy (L-CMD) and Werner's syndrome.
  15. A small nucleic acid molecule, characterized in that said small nucleic acid molecule comprises the polynucleotide according to any one of claims 1-6 or claim 7.
  16. The small nucleic acid molecule according to claim 15, characterized in that said small nucleic acid molecule is at least one selected from siRNA, ASO, shRNA, and miRNA.
  17. The nucleic acid sequence of claim 14, characterized in that ,
  18. A method for preventing and/or treating a disease associated with a lamin A mutation, characterized in that the method comprises the step of: administering a therapeutically effective amount of the polynucleotide according to any one of claims 1 to 6, or the pharmaceutical composition according to claim 8, or the small nucleic acid molecule according to any one of claims 15-17 to a subject.
  19. A non-therapeutic in vitro method for reducing the expression level of a lamin A mutant, or reducing the number of senescence-positive cells, or reducing the number of cells with dysmorphic nuclei, characterized in that the method comprises the step of: contacting a biological tissue in a non-therapeutic manner in vitro with the polynucleotide according to any one of claims 1 to 6, or the pharmaceutical composition according to claim 8, or the small nucleic acid molecule according to any one of claims 15-17.
  20. The method according to claim 19, characterized in that said biological tissue is a population of cardiomyocytes derived from progeria patients.

Description

This application claims priority to Chinese Patent Application No. CN202310688123.8, filed on June 9, 2023, entitled "Polynucleotide, Pharmaceutical Composition and Uses Thereof". TECHNICAL FIELD The present application relates to the field of biomedicine, and specifically to a polynucleotide, a pharmaceutical composition and uses thereof having efficacy in the treatment of certain diseases. BACKGROUND Lamin A protein is a component of the nuclear lamina structure and plays an important role in nuclear structure and function. Mutations in the LMNA gene can disrupt peripheral chromatin with specific epigenetic and molecular characteristics, leading to the erroneous expression of genes normally expressed in other cell types, causing congenital diseases such as progeria, familial dilated cardiomyopathy, muscular dystrophy, and lipodystrophy syndrome. Hutchinson-Gilford Progeria Syndrome, also known as childhood progeria, is a rare and fatal genetic disease. The incidence is approximately one in every 4 to 8 million newborns worldwide. HGPS is characterized by an accelerated aging process that is 5 to 10 times faster than normal. Patients exhibit a premature aging appearance resembling elderly individuals, accompanied by rapid systemic organ deterioration and a decline in physiological functions. Clinical symptoms include short stature, alopecia (hair loss), and delayed tooth eruption. Most patients die between the ages of 7 and 20, primarily from age-related diseases, such as cardiovascular disease. Existing studies have documented the possible cause of progeria, which is caused by a mutation in the LMNA gene encoding lamin A that results in a 150-nucleotide deletion in exon 11 containing the cleavage site for the zinc metalloproteinase 24 (ZMPSTE24) enzyme, preventing the normal cleavage of the farnesylated C-terminus of Lamin A. This leads to the production of a permanently farnesylated, toxic protein called progerin, which is the cause of the gradual degeneration of cell structure and function in children with progeria. Mutations occurring in the ZMPSTE24 gene can also lead to the formation of permanently farnesylated Lamin A protein, causing similar aging symptoms. Currently, Lonafarnib (trade name: Zokinvy) is the only FDA-approved drug for progeria. Lonafarnib is an oral farnesyltransferase inhibitor involved in protein isoprenylation modification. By inhibiting the isoprenylation of progerin, Lonafarnib can reduce the accumulation of progerin in the nucleus. However, clinical studies indicate that Lonafarnib does not restore a normal lifespan and is associated with safety concerns. Therefore, there remains a need for safer and more effective drugs for treating progeria. SUMMARY OF THE INVENTION To address and/or overcome the technical problems existing in the background art, an object of the present application is to provide a polynucleotide. Another object of the present application is to provide a pharmaceutical composition. Another object of the present application is to provide uses of the aforementioned polynucleotide or pharmaceutical composition. Another object of the present application is to provide a method for preventing and/or treating diseases associated with lamin A mutations. To achieve the above technical objectives, a first aspect of the present application provides a polynucleotide, said polynucleotide comprising a polynucleotide fragment complementary to the following sequence or a fragment thereof: (i) the sequence shown in SEQ ID NO: 11; and/or(ii) a polynucleotide sequence formed by substitution, deletion, or addition of one or several nucleotides in the sequence shown in SEQ ID NO: 11. In some preferred embodiments, said polynucleotide comprises a polynucleotide fragment complementary to the following sequence or a fragment thereof: (i) the sequence shown in SEQ ID NO: 1; and/or(ii) a polynucleotide sequence formed by substitution, deletion, or addition of one or several nucleotides in the sequence shown in SEQ ID NO: 1. In some preferred embodiments, said polynucleotide comprises a polynucleotide fragment complementary to the following sequence or a fragment thereof: (i) the sequence shown in SEQ ID NO: 10; and/or(ii) a polynucleotide sequence formed by substitution, deletion, or addition of one or several nucleotides in the sequence shown in SEQ ID NO: 10. In some preferred embodiments, said polynucleotide comprises at least one polynucleotide fragment selected from the group consisting of: (i) the polynucleotide fragments shown in SEQ ID NOs: 2-9; and(ii) polynucleotide fragments having at least 80% sequence identity with SEQ ID NOs: 2-9. In some preferred embodiments, said polynucleotide is at least one selected from the group consisting of: (i) the polynucleotides shown in SEQ ID NOs: 2-9; and(ii) polynucleotides having at least 80% sequence identity with SEQ ID NOs: 2-9. In some preferred embodiments, said polynucleotide is the polynucleotide shown in SEQ ID NO: 5 or the pol