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CN-122003502-A - Antisense oligonucleotides for treating kandelian disease

CN122003502ACN 122003502 ACN122003502 ACN 122003502ACN-122003502-A

Abstract

The present invention relates to antisense oligonucleotides (oligomers) complementary to NAT8L (N-acetyltransferase 8-like) pre-mRNA, which are capable of inhibiting NAT8L expression in cells expressing NAT 8L. Inhibition of NAT8L expression is beneficial in the treatment of kandelian disease (CANAVAN DISEASE).

Inventors

  • T. Freeland
  • M. Lebeck Johnson
  • S. Westergaard Rasmussen

Assignees

  • 康特拉医药股份有限公司

Dates

Publication Date
20260508
Application Date
20240927
Priority Date
20240328

Claims (8)

  1. 1. An antisense oligonucleotide selected from the following antisense oligonucleotides: a) AlxAlxCdxAlxEmxTdxTdxTdxAdxEdxGdxTdxCdxTmxGlxTdxEmxTl (SEQ ID NO: 3,ASO ID 66_707), b) TlxEmxTmxGdxTmxCdxTdxGdxTdxAdxTdxAdxTdxTdxEmoElxAl (SEQ ID NO: 1,ASO ID 66_130), c) TlxElxUoxGdxTmxCdxTdxGdxTdxAdxTdxAdxTdxTdxCdxEmxAl (SEQ ID NO: 2, ASO ID 66_124), and d) AmxAmxElxAmoEmxTdxTdxTdxAdxEdxGdxTdxCdxTmoGmxTlxEmxTm (SEQ ID NO: 4,ASO ID 66_709), Wherein Adx represents 2 '-deoxyadenosine-3' -phosphorothioate Aox represents 2 '-O-methyladenosine-3' -thiophosphate Amx represents 2 '-O-methoxyethyl adenosine-3' -thiophosphate Alx represents 2 '-O-beta-D-oxy LNA adenosine-3' -thiophosphate Cdx represents 2 '-deoxycytidine-3' -phosphorothioate Cox represents 2 '-O-methylcytidine-3' -phosphorothioate Cmx represents 2 '-O-methoxyethyl-5-methylcytidine-3' -phosphorothioate Clx represents 2 '-O-beta-D-oxoLNA-5-methylcytidine-3' -phosphorothioate Edx represents 2 '-deoxy-5-methylcytidine-3' -phosphorothioate Emx represents 2 '-O-methoxyethyl-5-methylcytidine-3' -phosphorothioate Elx represents 2 '-O-beta-D-oxoLNA-5-methylcytidine-3' -phosphorothioate Gdx represents 2 '-deoxyguanosine-3' -phosphorothioate Gox represents 2 '-O-methylguanosine-3' -phosphorothioate Gmx represents 2 '-O-methoxyethyl guanosine-3' -phosphorothioate Glx represents 2 '-O-beta-D-oxy LNA guanosine-3' -phosphorothioate Tdx represents 2 '-deoxythymidine-3' -phosphorothioate Tmx represents 2 '-O-methoxyethyl thymidine-3' -thiophosphate Tlx represents 2 '-O-beta-D-oxy LNA thymidine-3' -phosphorothioate Uox represents 2 '-O-methyluridine-3' -phosphorothioate Ado stands for 2 '-deoxyadenosine-3' -phosphate Aoo represents 2 '-O-methyladenosine-3' -phosphate Amo represents 2 '-O-methoxyethyl adenosine-3' -phosphate Alo represents 2 '-O-beta-D-oxy LNA adenosine-3' -phosphate Cdo represents 2 '-deoxycytidine-3' -phosphate Coo represents 2 '-O-methylcytidine-3' -phosphate Cmo represents 2 '-O-methoxyethyl-5-methylcytidine-3' -phosphate Edo represents 2 '-deoxy-5-methylcytidine-3' -phosphate Emo represents 2 '-O-methoxyethyl-5-methylcytidine-3' -phosphate Elo represents 2 '-O-beta-D-oxy LNA-5-methylcytidine-3' -phosphate Gdo represents 2 '-deoxyguanosine-3' -monophosphate Goo represents 2 '-O-methylguanosine-3' -monophosphate Gmo represents 2 '-O-methoxyethyl guanosine-3' -monophosphate Glo stands for 2 '-O-beta-D-oxy LNA guanosine-3' -phosphate Tdo represents 2 '-deoxythymidine-3' -phosphate Tmo represents 2 '-O-methoxyethyl thymidine-3' -phosphate Tlo represents 2 '-O-beta-D-oxy LNA thymidine-3' -phosphate Uoo represents 2 '-O-methyluridine-3' -monophosphate Ad stands for 2' deoxyadenosine Ao represents 2' -O-methyladenosine Am represents 2' -O-methoxyethyl adenosine Al represents 2' -O-beta-D-oxy LNA adenosine Cd represents 2' deoxycytidine Co represents 2' -O-methylcytidine Ed represents 2' -deoxy-5-methylcytidine Em represents 2' -O-methoxyethyl-5-methylcytidine El represents 2' -O-beta-D-oxoLNA-5-methylcytidine Gd represents 2' deoxyguanosine Go represents 2' -O-methylguanosine Gm represents 2' -O-methoxyethyl guanosine Gl stands for 2' -O-beta-D-oxy LNA guanosine Td stands for 2' deoxythymidine Tm represents 2' -O-methoxyethyl thymidine Tl represents 2' -O-beta-D-oxy LNA thymidine, and Uo represents 2' -O-methyluridine.
  2. 2. Antisense oligonucleotides as shown in fig. 1, fig. 2, fig. 3 or fig. 4.
  3. 3. A conjugate comprising the antisense oligonucleotide of claim 1 or 2, and at least one conjugate moiety covalently linked to the antisense oligonucleotide.
  4. 4. A pharmaceutical composition comprising the antisense oligonucleotide of claim 1 or 2, the conjugate of claim 3, and a pharmaceutically acceptable diluent, solvent, carrier, salt and/or adjuvant.
  5. 5. An in vitro method of reducing NAT8L expression in a host cell expressing NAT8L, the method comprising administering the antisense oligonucleotide of claim 1 or 2, the conjugate of claim 3, or the pharmaceutical composition of claim 4 to the cell in an effective amount.
  6. 6. A method of treating kandelian disease comprising administering a pharmaceutically effective amount of the antisense oligonucleotide of claim 1 or 2, the conjugate of claim 3, or the pharmaceutical composition of claim 4.
  7. 7. An antisense oligonucleotide according to claim 1 or 2, a conjugate according to claim 3 or a pharmaceutical composition according to claim 4 for use in the treatment of kandelian disease.
  8. 8. Use of an antisense oligonucleotide according to claim 1 or 2, a conjugate according to claim 3 or a pharmaceutical composition according to claim 4 in the manufacture of a medicament for the treatment of kandelian disease.

Description

Antisense oligonucleotides for treating kandelian disease Technical Field The present invention relates to antisense oligonucleotides (oligomers) complementary to NAT8L (N-acetyltransferase 8-like) pre-mRNA, which antisense oligonucleotides are capable of inhibiting NAT8L expression in cells expressing NAT 8L. Inhibition of NAT8L expression is beneficial in the treatment of kandelian disease. Background Kandelian disease (CANAVAN DISEASE) is a recessive genetic vacuolated leukodystrophy caused by ASPA mutations. ASPA encodes aspartate acylase (aspartoacylase), an oligodendrocyte enzyme that is necessary to cleave the abundant brain amino acid N-acetyl-L-aspartate (NAA) into acetate and L-aspartate. Kandelian disease is a rare and fatal autosomal recessive degenerative disorder that can lead to progressive damage to nerve cells and leukoencephalopathy. It is one of the most common degenerative brain diseases in infancy. Kandelian disease belongs to a group of genetic diseases called leukodystrophies (leukodystrophy), which refer to defects in white matter development and growth. Early symptoms of kandelian disease, such as hypotonia, megacephalum, poor head control, usually manifest at 3 to 6 months of age. Affected infants exhibit delayed developmental milestones (e.g., sitting, standing), and often never develop walking ability. The progression of the disease can lead to a loss of previously acquired ability and other symptoms such as seizures and mental retardation can also occur during infancy. The prognosis of kandelian disease is poor and in most cases death occurs before the age of one hundred years. Canavalia disease is caused by mutation of the ASPA gene, which encodes an aspartate-acyltransferase, which is required for the conversion of N-acetyl-L-aspartate (NAA) to acetate and L-aspartate. Currently, there are two main hypotheses about the pathological processes that result therefrom. The "oligodendrocyte starvation" hypothesis suggests that the lack of functional aspartate acylase results in a deficiency of NAA-derived acetate, which is required for biosynthesis of myelin (white matter) and thus leads to myelin/white matter degeneration in patients. In contrast, the "NAA toxicity" hypothesis suggests that excessive NAA concentration in the brain of patients lacking aspartate acylase leads to impaired osmotic homeostasis and thus triggers astrocyte and myelination, which is a hallmark of Canavalia disease (Pleasure et al , Neurochem Res. 2020 Mar;45(3):561-565. doi: 10.1007/s11064-018-2693-6. Epub 2018 Dec 8. Pathophysiology and Treatment of Canavan Disease.）. To support the "NAA toxicity" view, maier et al, 2015 reported that in Aspa deficient mice (Asap nur7/nur7, canavan disease model) mice lacking NAA synthase Nat8l (Aspa nur7/nur7/ Nat8l-/-) exhibited normal myelin content and no axonal degeneration, although NAA was not detected. However, survival was not improved (Maier et al , J Neurosci. 2015 Oct 28;35(43):14501-16. doi: 10.1523/JNEUROSCI.1056-15.2015. N-Acetylaspartate Synthase Deficiency Corrects the Myelin Phenotype in a Canavan Disease Mouse Model But Does Not Affect Survival Time）. furthermore, mice with only one complete Nat8l allele (Aspa nur7/nur7/ Nat8l-/+) accumulated less NAA, the disease pathology was also lighter, a phenotype improvement and normal longevity were shown Sohn et al （J Neurosci. 2017 Jan 11;37(2):413-421. doi: 10.1523/JNEUROSCI.2013-16.2016. Suppressing N-Acetyl-l-Aspartate Synthesis Prevents Loss of Neurons in a Murine Model of Canavan Leukodystrophy） also reported similar results. Subsequently Bannerman et al (2018) found that knocking down Nat8l expression in Aspa nur7/nur7 using an adeno-associated viral vector carrying a short hairpin Nat8l inhibitory RNA reduced NAA levels, inhibited the progression of disease pathology, and improved motor capacity (Bannerman et al , Brain Nat8l Knockdown Suppresses Spongiform Leukodystrophy in an Aspartoacylase-Deficient Canavan Disease Mouse Model Mol Ther. 2018 Mar 7;26(3):793-800. doi: 10.1016/j.ymthe.2018.01.002. Epub 2018 Jan 10）. Furthermore, hull et al (2020) showed that antisense oligonucleotides that reduced Nat8l expression in young adult Aspa nur7/nur7 mice also reduced NAA levels in Aspa nur7/nur7, reversed ataxia, and reduced cerebellar and thalamus vacuolation （Ann Neurol. 2020 Mar;87(3):480-485. doi: 10.1002/ana.25674. Epub 2020 Jan 22. Antisense Oligonucleotide Reverses Leukodystrophy in Canavan Disease Mice）. Taken together, these data suggest that inhibition of NAT8L and reduction of NAA levels may be a potentially effective therapeutic theory for the treatment of kandelian disease, a progressive and fatal neurodegenerative disease, for which no effective treatment is currently available. International patent application PCT/EP2023/077175, published as WO 2024/068997 A2, describes target regions for down-regulation of the NAT8L gene. Specifically, target regions capable of efficiently down-regulating gen