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WO2025202340A2 - Nouveaux oligonucléotides - Google Patents

Nouveaux oligonucléotides

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Publication number
WO2025202340A2
WO2025202340A2 PCT/EP2025/058355 EP2025058355W WO2025202340A2 WO 2025202340 A2 WO2025202340 A2 WO 2025202340A2 EP 2025058355 W EP2025058355 W EP 2025058355W WO 2025202340 A2 WO2025202340 A2 WO 2025202340A2
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WIPO (PCT)
Prior art keywords
oligonucleotide
seq
nucleotides
cell
senescence
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English (en)
Inventor
Lorna Wendy HARRIES
Benjamin Pereira LEE
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Senisca Ltd
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Senisca Ltd
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Publication of WO2025202340A2 publication Critical patent/WO2025202340A2/fr
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
    • C12N15/113—Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2310/00—Structure or type of the nucleic acid
    • C12N2310/10—Type of nucleic acid
    • C12N2310/11—Antisense
    • C12N2310/113—Antisense targeting other non-coding nucleic acids, e.g. antagomirs
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2310/00—Structure or type of the nucleic acid
    • C12N2310/30—Chemical structure
    • C12N2310/31—Chemical structure of the backbone
    • C12N2310/315—Phosphorothioates
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2310/00—Structure or type of the nucleic acid
    • C12N2310/30—Chemical structure
    • C12N2310/32—Chemical structure of the sugar
    • C12N2310/323—Chemical structure of the sugar modified ring structure
    • C12N2310/3233—Morpholino-type ring

Definitions

  • the invention relates to novel oligonucleotides and their use in inhibiting or reducing senescence.
  • the oligonucleotides target specific sequences in the 3' untranslated region (UTR) of heterogeneous nuclear ribonucleoprotein D HNRNPD') mRNA and blocks one or more inhibitory microRNA (miRNA) binding sites. This promotes the physiologically regulated expression of HNRNPD and reduces senescence.
  • UTR 3' untranslated region
  • miRNA inhibitory microRNA
  • HNRNPD also known as AU-rich element RNA-binding protein 1, AUF1 encodes heterogeneous nuclear ribonucleoprotein particle D, and usually but not exclusively inhibits splice site usage. It also has roles in RNA export, degradation of target transcripts (including proinflammatory cytokines) via its binding to A rich elements of target genes, and telomere maintenance.
  • Vyavahare et al. have shown that miR-141-3p (a miRNA) promotes inflammation and senescence. Direct inhibition of miR-141-3p reduces premature senescence in C2C12 cells when measured using the senescence markers pl6, p21 and senescence-associated beta galactosidase (SA-p-Gal) staining.
  • HNRNPD HNRNPD
  • AUF1 HNRNPD
  • HNRNPD HNRNPD
  • miR-141-3p has its antisenescence effect directly via HNRNPD (AUF1) since microRNAs act by binding to short degenerate sequences in the 3' untranslated region of their target genes and as such have thousands of predicted targets.
  • HNRNPD HNRNPD
  • miR-141-3p has over 1000 such targets (https://mirdb.orq/) of which HNRNPD AUF1) is just one.
  • the inhibitor of miR-141-3p used in the study was therefore not specific to HNRNPD AU Fl in the sense it inactivates the activity of miR-141-3p in its entirety and thus regulates all of its target genes.
  • the oligonucleotides of the invention specifically target the interaction between HNRNPD and its upstream regulators miR-141-3p and miR-146b-5p and do not influence the effect of the inhibitory miRNA on other genes. This greatly avoids the risk of off-target side effects and potentially increases the safety of the oligonucleotides of the invention. This is also a key difference from the approach described by Vyavahare et al. ("Inhibiting MicroRNA-141-3p Improves Musculoskeletal Health in Aged Mice.” Aging and Disease, Vol. 14 ,6 2303-2316. 21 Apr. 2023, doi: 10.14336/AD.2023.0310-1), where the authors target miR-141-3p itself.
  • Another advantage of the specific targeting of the invention is that it is not necessary to introduce supraphysiological levels of the HNRNPD target because its expression remains governed by the usual regulatory controls of the cell. It is important to maintain the endogenous regulation of HNRNPD to avoid it becoming an oncogene.
  • the invention provides a method of reducing senescence in a cell, comprising contacting the cell with an oligonucleotide which specifically hybridises to a target portion of the 3' untranslated region (UTR) of heterogeneous nuclear riboprotein particle D HNRNPD) mRNA and blocks at least one inhibitory microRNA (miRNA) binding site and thereby reducing senescence.
  • an oligonucleotide which specifically hybridises to a target portion of the 3' untranslated region (UTR) of heterogeneous nuclear riboprotein particle D HNRNPD) mRNA and blocks at least one inhibitory microRNA (miRNA) binding site and thereby reducing senescence.
  • the invention also provides: an oligonucleotide which specifically hybridises to a portion of the 3' UTR of HNRNPD mRNA and blocks at least one inhibitory miRNA binding site for use in a method of reducing senescence in a cell in a subject; an oligonucleotide which specifically hybridises to a portion of the 3' UTR of HNRNPD mRNA and blocks at least one inhibitory miRNA binding site for use in a method of treating or preventing one or more signs of ageing in a subject; an oligonucleotide of about 50 or fewer nucleotides and comprising or consisting of any one of the sequences shown in SEQ ID NOs: 1-28, 33-50, and 55-74; and a pharmaceutical composition comprising an oligonucleotide of the invention and a pharmaceutically acceptable carrier and/or diluent.
  • FIG. 1 SOLOOl, SOL002 or SOL003 causes a reduction in CDKN2A (pl6) expression in senescent primary lung fibroblasts.
  • SOLOOl, SOL002 and SOL003 were initial designs to HNRNPD, designed to block an autoregulatory feedback loop that exists between HNRNPD and its upstream regulators miR-141-3p and /or miR-146b-5p. These were morpholino oligonucleotides (PMOs), introduced to senescent human primary lung fibroblast cells at different concentrations using lipofectamine. Both oligos produced a reduction in senescent cell load under these conditions at multiple concentrations as measured by pl6 expression.
  • PMOs morpholino oligonucleotides
  • SOL044, SOL045 or SOL046 caused a reduction in SA-p-Gal activity, a reduction in pl6 expression and a decrease in DNA damage load in senescent primary lung fibroblasts.
  • SOL044 SOL045 and SOL046 were 2'-O-(2-Methoxyethyl)-oligoribonucleotides (2'-MOE) equivalents of SOLOOl, SOL002 and SOL003, delivered to senescent lung fibroblasts by lipofectamine delivery. These had a different chemistry but brought about a marked reduction in senescence as assessed by SA-p-Gal staining, a reduction in DNA damage load and a small effect on proliferation for SOL046.
  • 2'-MOE 2-Methoxyethyl-oligoribonucleotides
  • FIG. 4 SOL044, SOL045 or SOL046 caused decreases in markers of fibrosis and increased expression of markers protective against fibrosis in senescent primary human lung cells.
  • the primary aim was to design and validate a portfolio of oligonucleotide drugs for idiopathic pulmonary fibrosis (IPF).
  • IPF idiopathic pulmonary fibrosis
  • the inventors sought to determine whether SOL044, SOL045 and SOL046 could impact on fibrotic phenotypes in senescent primary human lung fibroblasts assessed by SA-p-Gal staining.
  • Oligonucleotides were delivered to cells by lipofectamine. All three oligonucleotides were able to reduce markers of fibrosis and increase protective anti-fibrotic markers.
  • FIG. 5 SOL044, SOL045 or SOL046 cause a decrease in markers of senescence in human primary dermal fibroblasts when delivered via gymnosis.
  • Gymnosis is the delivery of naked oligonucleotides into cells without a lipid nanoparticle carrier. These data show it was possible to deliver the oligonucleotides to human cells in vitro in this fashion, and still observe efficacy.
  • Senescent dermal fibroblasts exposed to a 2'-MOE oligonucleotide tagged with a fluorochrome for 48 hours show dose-dependent uptake and retention of oligonucleotide.
  • B Untagged SOL044, SOL045 and SOL046 delivered to senescent primary human lung fibroblasts by gymnosis caused a reduction in senescence as by SA-p-Gal staining.
  • FIG. 6 SOL044, SOL045 or SOL046 delivered by gymnosis caused a decrease in markers of senescence in human primary lung fibroblasts. SOL044, SOL045 and SOL046 maintain their effect on SA-p-Gal activity in senescent primary human lung fibroblasts when delivered by gymnosis.
  • FIG. 7 SOL045 or SOL046 caused a decrease in markers of senescence and inflammation in human primary lung fibroblasts from patients with idiopathic pulmonary fibrosis.
  • SOL045 and SOL046 oligonucleotides delivered by lipofectamine reduced senescent cell load, caused cells to re-enter cell cycle and reduced DNA damage burden.
  • SOL045 also attenuated the expression of LIF and IL6 in human primary lung fibroblasts from a 55yr old male patient with IPF. Senescence was assessed by SA-p-Gal staining.
  • FIG. 8 SOL045 or SOL046 delivered by oligofectamine caused a decrease in markers of senescence that lasts at least 10 days in primary lung fibroblasts from patients with idiopathic pulmonary fibrosis. Fibroblasts from a 55-yr old male IPF patient maintained effects on senescence phenotypes at 5-days and 10-days post treatment, after treatment with SOL045 or SOL046 delivered by lipofectamine. Senescence was assessed by SA-p-Gal staining.
  • FIG. 9 SOL044, SOL045 or SOL046 delivered by gymnosis caused a decrease in fibrotic markers compared to standard IPF treatments in primary fibroblasts from IPF patient cells.
  • DMSO DMSO carrier.
  • Primary fibroblasts from a 55yr old male IPF patient were treated with the gold standard clinical IPF treatments, pirfenidone and nintedanib or with SOL044 - SOL046, delivered by gymnosis.
  • the stains shown are for protein expression of either (1) oSMA, a marker of fibroblast to myofibroblast transdifferentiation or (2) Collagen 1, an important marker of fibrosis.
  • the oligonucleotides had at least comparable effects on fibrotic markers as the state-of-the-art drugs in our hands.
  • FIG. 10 SOL045 delivered by gymnosis reduces the protein expression of MMP7 and GREMLIN1 as measured by ELISA.
  • MMP7 is an important predictive and prognostic marker of IPF
  • GREMLIN 1 is the initiating factor in the fibrotic cascade.
  • FIG 11 SOL045 caused a decrease in senescence, an increase in ACAN (a marker of cartilage health) and a decrease in MMP13, a marker of cartilage remodelling in senescent articular chondrocytes from a patient with osteoarthritis.
  • FIG. 12 SOL044, SOL045 or SOL046 caused a reduction of markers of senescence in senescent primary human retinal endothelial cells.
  • FIG. 13 SOL044, SOL045 or SOL046 caused a reduction of markers of senescence in senescent primary human pigmented retinal epithelial cells.
  • FIG. 14 SOL045, SOL068 and SOL002 (variants of the same oligonucleotide sequence) delivered intratracheally to C57BL6/J mice using a single dose of lOmg/kg at Day 0, showed chemistry-dependent pharmacokinetic profiles.
  • A Full MOE-PS oligonucleotide SOL045 showed the most favourable PK profile, with a characteristic slow accumulation in the lungs up to 48h, followed by a drop at 72h and a second slow accumulation to 168h.
  • B Alternating MOE/DNA-PS oligonucleotide SOL068 was rapidly released from lungs after 24h.
  • FIG. 15 SOL044-4, SOL045+5, SOL045+8 and scrambled control (SCR) oligonucleotides delivered intratracheally to C57BL6/J mice using a single dose of lOmg/kg at Day 0, showed favourable pharmacokinetic profiles.
  • SOL044-4 oligonucleotide was cleared from the lungs over the timecourse, but remained detectable at low levels 168h post-dose.
  • SOL045+5 oligonucleotide was cleared from the lungs over the timecourse, but remained detectable at reasonable levels 168h post-dose.
  • Figure 16 A subset of SOL044 oligonucleotide panel delivered gymnotically at a dose of 300nM using a low-throughput methodology confirmed that SOL044 series oligonucleotides have significant effect on markers of senescence in human primary lung fibroblasts from an IPF patient, showing a reduction in senescent cell load of up to ⁇ 44%. (Key - SCR: scrambled control oligonucleotide)
  • Figure 17 A subset of SOL045 oligonucleotide panel delivered gymnotically at a dose of 300nM using a low-throughput methodology confirmed that SOL045 series oligonucleotides have significant effect on markers of senescence in human primary lung fibroblasts from an IPF patient, showing reductions in senescent cell load of up to ⁇ 45%. (Key - SCR: scrambled control oligonucleotide)
  • FIG. 18 SOL045 oligonucleotide delivered gymnotically at doses of 200nM and 400nM has a significant effect of up to ⁇ 54% reduction in markers of senescence in human Precision- Cut Lung Slices (PCLS) from a patient with Chronic Obstructive Pulmonary Disease (COPD).
  • PCLS Precision- Cut Lung Slices
  • COPD Chronic Obstructive Pulmonary Disease
  • FIG 19 SOL045 oligonucleotide delivered gymnotically at a dose of 400nM has a significant effect on reduction of oSMA (a marker of fibroblast-to-myofibroblast transdifferentiation) in human Precision-Cut Lung Slices (PCLS) from a patient with Chronic Obstructive Pulmonary Disease (COPD).
  • PCLS Precision-Cut Lung Slices
  • COPD Chronic Obstructive Pulmonary Disease
  • FIG. 20 SOL045 oligonucleotide delivered gymnotically at a dose of 200nM has a significant effect of up to -44% reduction in MMP7 (a marker of pathological tissue remodelling) in human Precision-Cut Lung Slices (PCLS) from a patient with Chronic Obstructive Pulmonary Disease (COPD) at both Day 11 (A) and Day 14 (B) in culture.
  • MMP7 a marker of pathological tissue remodelling
  • PCLS Precision-Cut Lung Slices
  • COPD Chronic Obstructive Pulmonary Disease
  • FIG. 21 SOL045 oligonucleotide delivered gymnotically at a dose of 400nM has a significant effect of up to -80% reduction in markers of senescence in human Precision-Cut Lung Slices (PCLS) from a patient with non-IPF Interstitial Lung Fibrosis (ILF).
  • PCLS Precision-Cut Lung Slices
  • IVF Interstitial Lung Fibrosis
  • FIG 22 SOL045 oligonucleotide delivered gymnotically at a dose of 400nM has a significant effect of up to -74% reduction in markers of senescence in human Precision-Cut Lung Slices (PCLS) from a patient with Idiopathic Pulmonary Fibrosis (IPF).
  • PCLS Precision-Cut Lung Slices
  • IPF Idiopathic Pulmonary Fibrosis
  • Figure 23 Lead oligonucleotides SOL044-4, SOL045+5 and SOL045+8 delivered gymnotically at a dose of 400nM have significant effects of up to -55% reduction in markers of senescence in human Precision-Cut Lung Slices (PCLS) from a patient with Chronic Obstructive Pulmonary Disease (COPD).
  • PCLS Precision-Cut Lung Slices
  • COPD Chronic Obstructive Pulmonary Disease
  • FIG. 24 Lead oligonucleotides SOL044-4, SOL045+5 and SOL045+8 delivered gymnotically at a dose of 400nM have significant effects of up to -59% reduction in MMP7 (a marker of pathological tissue remodelling) in human Precision-Cut Lung Slices (PCLS) from a patient with Chronic Obstructive Pulmonary Disease (COPD) at both Day 11 (A) and Day 14 (B) in culture.
  • PCLS Precision-Cut Lung Slices
  • COPD Chronic Obstructive Pulmonary Disease
  • FIG. 25 Lead oligonucleotides SOL044-4, SOL045+5 and SOL045+8 delivered gymnotically at a dose of 400nM have significant effects of up to -60% reduction in MMP9 (a marker of pathological tissue remodelling) in human Precision-Cut Lung Slices (PCLS) from a patient with Chronic Obstructive Pulmonary Disease (COPD) at both Day 11 (A) and Day 14 (B) in culture.
  • PCLS Precision-Cut Lung Slices
  • COPD Chronic Obstructive Pulmonary Disease
  • Figure 26 Lead oligonucleotides SOL044-4, SOL045+5 and SOL045+8 delivered gymnotically at a dose of 400nM have significant effects of up to ⁇ 64% reduction in GM- CSF (a marker of fibrogenesis) in human Precision-Cut Lung Slices (PCLS) from a patient with Chronic Obstructive Pulmonary Disease (COPD) at both Day 11 (A) and Day 14 (B) in culture.
  • PCLS Precision-Cut Lung Slices
  • COPD Chronic Obstructive Pulmonary Disease
  • heterogeneous nuclear ribonucleoprotein particle D is interchangeable with “heterogeneous nuclear ribonucleoprotein D”.
  • a variant of SEQ ID NO: 1 having at least "about” 90% identity or homology to the sequence of SEQ ID NO: 1 over its entire length includes a variant of SEQ ID NO: 1 having at least 80% identity or homology to the sequence of SEQ ID NO: 1 over its entire length.
  • the invention provides a method of reducing senescence in a cell.
  • Senescence is associated with a variety of general traits, including cell cycle arrest and the senescence-associated secretory phenotype (SASP), an enlarged and flattened morphology, expanded lysosomal compartment and particular chromatin and epigenetic alterations.
  • SASP senescence-associated secretory phenotype
  • Senescence can be measured using routine methods in the art, including those described in Gonzalez-Gualda, Estela et al. "A guide to assessing cellular senescence in vitro and in vivo.”
  • a reduction in senescence in the cell can be measured in a variety of ways including, but not limited to, re-entry to the cell cycle, structural changes, a reduction in pro-survival pathways, a reduction in the SASP and plasma membrane protein expression.
  • a reduction in senescence in the cell can be measured in a variety of ways including, but not limited to, reentry to the cell cycle, structural changes, a reduction in pro-survival pathways, a reduction in the number of cells staining positive for the senescence-associated biochemical marker senescence-associated beta galactosidase (SA-p-Gal, or SAB), a reduction in the SASP and plasma membrane protein expression.
  • SA-p-Gal biochemical marker senescence-associated beta galactosidase
  • Re-entry to the cell cycle can be identified by measuring DNA synthesis, for instance by measuring increased BrdU and EdU, an increased proliferation, for instance by measuring increased Ki67, an inhibition of the pl6-pRB axis, for instance by measuring decreased pl6INK4a and pRB and increased phospho-pRb, an inhibition of the p53-p21 axis, for instance by measuring decreased p21, p53, phospho-p53, DECI (BHLHB2) and PPP1A.
  • DNA synthesis for instance by measuring increased BrdU and EdU
  • an increased proliferation for instance by measuring increased Ki67
  • an inhibition of the pl6-pRB axis for instance by measuring decreased pl6INK4a and pRB and increased phospho-pRb
  • an inhibition of the p53-p21 axis for instance by measuring decreased p21, p53, phospho-p53, DECI (BHLHB2) and PPP1A.
  • Structural changes associated with reduced senescence include, reversal of the enlarged, flattened shape, decreased lysosomal compartment and activity, for instance by measuring decreased SA-p-galactosidase, SA-o-Fucosidase and Lipofuscin, reversal of DNA damage, for instance by measuring decreased yH2AX, 53BPI, Rad 17, ATR, ATM, MDC1 and TIF, reduced reactive oxygen species (ROS), increased telomere length, a reduction in senescence-associated heterochromatic foci (SAHFs), for instance by measuring decreased DAPI/Hoechst 33342, HIRA, H3K9-methylation, PML bodies and HPl-gamma, and increased Lamin Bl in the nuclear membrane.
  • ROS reduced reactive oxygen species
  • SAHFs senescence-associated heterochromatic foci
  • a reduction in pro-survival pathways can be measured by the presence of Annexin V, cleaved poly (ADP-ribose) polymerase (PARP), cleaved caspase 2/3/9 and TUNEL (terminal deoxynucleotidyl transferase dUTP nick end labelling) staining.
  • Annexin V cleaved poly (ADP-ribose) polymerase (PARP)
  • PARP cleaved caspase 2/3/9
  • TUNEL terminal deoxynucleotidyl transferase dUTP nick end labelling
  • a reduction in the SASP can be measured by measuring a decrease in a number of cytokines, including interleukin-6 (IL-6), IL-8, CXC motif chemokine receptor 2 (CXCR2), insulin-like growth factor 2 (IGF2), insulin-like growth factor-binding protein 3 (IGFBP3), IGFBP5, IGFBP7, stanniocalcin-1 (STC1), growth differentiation factor (GDF15) and serine protease inhibitors (SERPINs).
  • IL-6 interleukin-6
  • CXCR2 CXC motif chemokine receptor 2
  • IGF2 insulin-like growth factor 2
  • IGFBP3 insulin-like growth factor-binding protein 3
  • IGFBP5 insulin-like growth factor-binding protein 3
  • STC1 growth differentiation factor
  • GDF15 growth differentiation factor
  • SERPINs serine protease inhibitors
  • a reduction in the SASP can be measured by measuring a decrease in a number of cytokines.
  • cytokines may, for example, include interleukin-6 (IL-6), interleukin-8 (IL- 8), interleukin-1 alpha (IL-lo), interleukin-1 beta (IL-1B or IL-ip), interleukin-2 (IL-2), CXC motif chemokine receptor 2 (CXCR2), CXC motif ligand 1 (CXCL1), insulin-like growth factor 2 (IGF2), insulin-like growth factor-binding protein 2 (IGFBP2), insulin-like growth factorbinding protein 3 (IGFBP3), insulin-like growth factor-binding protein 4 (IGFBP4), insulinlike growth factor-binding protein 5 (IGFBP5), insulin-like growth factor-binding protein 6 (IGFBP6), insulin-like growth factor-binding protein 7 (IGFBP7), stanniocalcin-1 (STC1), growth differentiation factor (GDF15), matrix metallo
  • the reduction in senescence is preferably identified as described in the Example.
  • the reduction in senescence is preferably identified by measuring a reduction in pl6 expression. This is shown in Figures 1 and 2.
  • the reduction in senescence is preferably identified by measuring a reduction in SA-p-Gal staining. This is shown in Figures 3-7.
  • the reduction in senescence is preferably identified by measuring a reduction in expression of one or more of pl6, p21 and p53, such as a reduction in expression of pl6, a reduction in expression of p21, a reduction in expression of p53, a reduction in expression of pl6 and p21, a reduction in expression of p21 and p53, a reduction in expression of pl6 and p53, or a reduction in expression of pl6, p21 and p53.
  • the reduction in senescence is preferably identified by measuring a reduction in SA-p-Gal (SAB) staining, alone or in combination with an assessment of morphology, for example cell size, nuclear size and/or granularity.
  • SAB SA-p-Gal
  • the senescence may be reduced by any amount.
  • the senescence is preferably reduced by at least about 2%, at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, or at least about 100%.
  • Senescence can be measured as discussed above.
  • the amount of senescence is typically reduced in the method of the invention compared with the amount of senescence in the cell before it is contacted with the oligonucleotide.
  • the cell used in the method of the invention preferably comprises increased senescence before it is contacted with the oligonucleotide.
  • the invention provides a method of reducing senescence in a cell which comprises increased senescence or an increased amount of senescence.
  • Senescence is typically increased compared with a normal or healthy cell of the same type. The association of an increases in senescence with diseases or conditions are discussed in more detail below.
  • HNRNPD physiologically regulated expression of HNRNPD.
  • the expression of HNRNPD may increase.
  • the expression of HNRNPD may decrease.
  • the expression of HNRNPD may remain about the same.
  • it is extremely difficult to demonstrate a directional effect on HNRNPD expression levels following treatment in vitro because natural regulated HNRNPD expression is labile and responsive to multiple stimuli.
  • Contacting the cell with the oligonucleotide in accordance with the invention preferably promotes the physiologically regulated expression of HNRNPD and thereby reduces senescence.
  • the cell may be any type of cell.
  • the cell is typically a eukaryotic cell.
  • the cell may be a protozoan, algal, fungal, plant or animal cell.
  • the cell is preferably mammalian.
  • the cell is preferably a human, dog, cat, primate, horse, murine, rat, rodent, bovine, murine, porcine, or ovine cell.
  • the cell is preferably a human cell.
  • the cells may be a plant cell, such as a cereal, legume, fruit, or vegetable cells. Examples include, but are not limited to, wheat, barley, oat, canola, maize, soya, rice, banana, apple, tomato, potato, grape, tobacco, bean, lentil, sugar cane, cocoa, cotton, tea, or coffee cell.
  • the animal or mammalian cell may be derived from the ectoderm, endoderm, or mesoderm.
  • the cell may be an endothelium cell.
  • the cell may be an epithelium cell.
  • the cell may be derived from immune system, heart, brain, vasculature, skin, intestine, lung, thyroid, reproductive organ, bladder, kidney, pancreas, oral mucosal, eye or liver.
  • the cell may be a stem cell, such as an embryonic stem cell, induced pluripotent stem cell or mesenchymal stem cell, bone cell, such as an osteoclast, osteoblast or osteocyte, tendon cell, such as a tenoblast or tenocyte, chondrocyte, synovial cell, vascular cell, connective tissue cell, such as a fibroblast, blood cell, such as a red blood cell, immune cell, platelet, neutrophil or basophil, muscle cells, such as a skeletal muscle cell, cardiac muscle cell or smooth muscle cell, reproductive cell, such as a sperm, oocyte, duct cell or epididymal cell, secretory cell, adipocyte, liver lipocyte, epithelial cell, odontoblast, cementoblast, hormone-secreting cell, barrier cell, exocrine secretory epithelial cell, nerve cell, astrocyte, oligodendrocyte, or neuron.
  • stem cell such as an embryonic stem cell, induced pluripotent
  • An oligonucleotide is a polymer comprising two or more nucleotides.
  • the nucleotides can be naturally occurring or artificial.
  • a nucleotide typically contains a nucleobase, a sugar and at least one linking group, such as a phosphodiester, phosphoramidate, phosphorodiamidate, methylphosphonate or phosphorothioate group.
  • the linking groups form internucleoside linkages.
  • Preferred nucleosides include, but are not limited to, adenosine, guanosine, 5-methyluridine, uridine, cytidine, 5-methyl cytidine, deoxyadenosine, deoxyguanosine, thymidine, deoxyuridine and deoxycytidine.
  • the nucleosides are preferably adenosine, guanosine, thymidine, and 5-methyl cytidine.
  • the nucleotides in the polynucleotide may be attached to each other in any manner.
  • the nucleotides may be linked by phosphodiester, phosphoramidate, phosphorodiamidate, methylphosphonate or phosphorothioate linkages.
  • the nucleotides are typically attached by their sugar and phosphate groups as in nucleic acids.
  • the nucleotides may be connected via their nucleobases as in pyrimidine dimers.
  • the oligonucleotide may be ribonucleic acid (RIMA) or deoxyribonucleic acid (DNA).
  • RIMA ribonucleic acid
  • DNA deoxyribonucleic acid
  • the oligonucleotide may be modified in any of the ways discussed below.
  • the oligonucleotide may be any synthetic nucleic acid known in the art, such as peptide nucleic acid (PNA), glycerol nucleic acid (GNA), threose nucleic acid (TNA), locked nucleic acid (LNA), morpholino nucleic acid or other synthetic polymers with nucleotide side chains.
  • the oligonucleotide is preferably modified to (a) increase its binding affinity for the 3' UTR, (b) increase its resistance to one or more nucleases, (c) alter its pharmacokinetics and/or pharmacodynamics, (d) alter its tissue distribution, (e) increase its stability, (f) improve its toxicity, (g) increase its uptake and/or retention by a cell, (h) reduce its immunogenicity, (i) reduce its off-target effects, (j) alter its charge and/or hydrophobicity, (k) alter its protein binding, or any combination thereof.
  • the oligonucleotide is typically modified to reduce or lessen its toxicity.
  • the one or more nucleases may be one or more RNAses or DNAses.
  • the increases in affinity and/or resistance may be measured using routine methods.
  • the altered pharmacokinetics and/or tissue distribution may be measured using routine methods. Affinity is also discussed below in the context of hybridisation. Suitable modifications are described in, for example, US 20210079391A1 and US8183363B2.
  • references to "modification” or “modified” or the like typically refer to modified versions of RNA nucleotides, RNA, DNA nucleotides or DNA.
  • the oligonucleotide may comprise any of the modifications discussed above or below.
  • the one or more modified sugars may be substituted sugars.
  • the one or more modified sugars may be bicyclic or tricyclic sugars.
  • the one or more modified sugars may be one or more sugar surrogates.
  • Such sugar surrogates may comprise one or more substitutions corresponding to those of substituted sugars.
  • Modified sugars are substituted sugars comprising one or more substituents including, but not limited to, substituents at the 2', 3' and 5' positions, such as the 2' position, the 3' position, the 5' position, the 2' and 3' positions, the 2' and 5' positions, the 3' and 5' positions and the 2', 3' and 5' positions.
  • substituents at the 2', 3' and 5' positions such as the 2' position, the 3' position, the 5' position, the 2' and 3' positions, the 2' and 5' positions, the 3' and 5' positions and the 2', 3' and 5' positions.
  • nucleosides having modified sugar moiety include without limitations nucleosides comprising 5-vinyl, 5-methyl, 4'-S, 2'-F, 2'-OCH 3 (OMe or O-methyl), and 2'- O(CH 2 )2OCH 3 (2'-MOE) substituent groups.
  • Other examples of sugar substituents can be found in US 20210079391A1 and WO 2008/101157.
  • Bicyclic nucleotides include, but are not limited to, a-L-Methyleneoxy (4'-CH 2 -O-2') BNA, [3- D-Methyleneoxy (4'-CH 2 -O-2') BNA (also referred to as locked nucleic acid or LNA), Ethyleneoxy (4'-(CH 2 ) 2 -O-2') BNA, Aminooxy (4'-CH 2 -O-N(R)-2') BNA, Oxyamino (4'-CH 2 - N(R)-O-2') BNA, Methyl(methyleneoxy) (4'-CH(CH 3 )-O-2') BNA (also referred to as constrained ethyl or cEt), methylene-thio (4'-CH 2 -S-2') BNA, methylene-amino (4'-CH 2 - N(R)
  • substituted sugars comprise one or more non-bridging sugar substituent and one or more bridging sugar substituent (e.g., 5'-substituted and 4'-2' bridged sugars) as disclosed in WO 2007/134181, wherein LNA is substituted with, for example, a 5'-methyl or a 5'-vinyl group).
  • bridging sugar substituent e.g., 5'-substituted and 4'-2' bridged sugars
  • carbocyclic bicyclic nucleotides having a 4'-2' bridge have been described (see, e.g., Freier et al., Nucleic Acids Research, 1997, 25(22), 4429-4443 and Albaek et al., J. Org. Chem., 2006, 71, 7731-7740).
  • Morpholinos may be modified, for example by adding or altering various substituent groups from the morpholino structure. Such sugar surrogates are referred to as "modified morpholinos.”
  • modified nucleobases or nucleobase mimetics can be incorporated into the oligonucleotide.
  • Modified nucleobases can include nucleobases with minimal modification from the parent nucleobase, such as 7-deaza purine, 5-methyl cytosine, or a G-clamp.
  • Modified nucleobases can also include nucleobase mimetics such as tricyclic phenoxazine nucleobase mimetics. Methods for preparation of the above noted modified nucleobases are well known to those skilled in the art.
  • the oligonucleotide preferably comprises one or more modified nucleobases.
  • the nucleotides in the oligonucleotide each comprise a nucleobase.
  • the one or more nucleobases are typically modified compared with the nucleobases in RIMA or DNA.
  • the inclusion of modified nucleobases may improve any of the biological properties discussed above as (a)-(k) and typically increases the binding affinity of the oligonucleotide for the 3' UTR.
  • the oligonucleotide may comprise one type of modified nucleobase.
  • the oligonucleotide may comprise two or more different types of modified nucleobases. Every nucleotide in the oligonucleotide may comprise a modified nucleobase.
  • the one or more nucleobase modifications are independent of the sequence of nucleobases.
  • Universal nucleotides are capable of base pairing with any nucleotide.
  • the universal nucleotide preferably comprises one of the following nucleobases: hypoxanthine, 4- nitroindole, 5-nitroindole, 6-nitroindole, formylindole, 3-nitropyrrole, nitroimidazole, 4- nitropyrazole, 4-nitrobenzimidazole, 5-nitroindazole, 4-aminobenzimidazole or phenyl (C6- aromatic ring).
  • the universal nucleotide more preferably comprises one of the following nucleosides: 2'-deoxyinosine, inosine, 7-deaza-2'-deoxyinosine, 7-deaza-inosine, 2-aza- deoxyinosine, 2-aza-inosine, 2-O'-methylinosine, 4-nitroindole 2’-deoxyribonucleoside, 4- nitroindole ribonucleoside, 5-nitroindole 2’ deoxyribonucleoside, 5-nitroindole ribonucleoside, 6-nitroindole 2’ deoxyribonucleoside, 6-nitroindole ribonucleoside, 3- nitropyrrole 2’ deoxyribonucleoside, 3-nitropyrrole ribonucleoside, an acyclic sugar analogue of hypoxanthine, nitroimidazole 2’ deoxyribonucleoside, nitroimidazole ribonucleoside,
  • the universal nucleotide more preferably comprises 2'- deoxyinosine.
  • the universal nucleotide is more preferably IMP or dIMP.
  • the universal nucleotide may be dPMP (2’-Deoxy-P-nucleoside monophosphate) or dKMP (N6-methoxy-2, 6-diaminopurine monophosphate).
  • nucleobases include, but are not limited to, tricyclic pyrimidines such as phenoxazine cytidine([5,4-b][l,4]benzoxazin-2(3H)-one), phenothiazine cytidine (lH-pyrimido[5,4-b][l,4]benzothiazin-2(3H)-one), G-clamps such as a substituted phenoxazine cytidine (e.g.
  • nucleobases may also include those in which the purine or pyrimidine base is replaced with other heterocycles, for example 7-deaza-adenine, 7-deazaguanosine, 2-aminopyridine and 2-pyridone.
  • nucleobases include those disclosed in U 3,687,808, The Concise Encyclopedia Of Polymer Science And Engineering, Kroschwitz, J. I., Ed., John Wiley & Sons, 1990, 858-859, Englisch et al., Angewandte Chemie , International Edition, 1991, 30, 613, Sanghvi, Y. S., Chapter 15, Antisense Research and Applications, Crooke, S. T. and Lebleu, B., Eds., CRC Press, 1993, 273-288.
  • cytosine containing nucleotides in an oligonucleotide are 5-methyl cytosine nucleotides.
  • the oligonucleotide preferably comprises one or more phosphorothioate (PS) internucleoside linkages and/or one or more phosphorodiamidate morpholino (PMO) internucleoside linkages.
  • the oligonucleotide preferably comprises only phosphorothioate (PS) internucleoside linkages or only phosphorodiamidate internucleoside linkages.
  • the oligonucleotide preferably comprises one of the following: 1. only phosphorodiamidate linked morpholino nucleotides;
  • the oligonucleotide preferably comprises alternating 2'-MOE nucleotides and DNA nucleotides with PS on the DNA nucleotides.
  • the oligonucleotide preferably comprises or consists of the sequence shown in any one of SEQ ID NOs: 1-28, 33-50, and 55-74 and having any of the modifications discussed above, especially those set out in #l-#4 in the previous paragraph.
  • the oligonucleotide preferably comprises or consists of the sequence shown in any one of SEQ ID NOs: 1-26, 33-50, and 55-74 and having any of the modifications discussed above, especially those set out in #1- #4 in the previous paragraph.
  • the oligonucleotide preferably comprises or consists of the sequence shown in any one of SEQ ID NOs: 1-2, 5-9. 12, 13, 15-18, 20-26, 33-36, 38-50 and 55-74 and having any of the modifications discussed above, especially those set out in #l-#4 in the previous paragraph.
  • the oligonucleotide preferably comprises or consists of the sequence shown in SEQ ID NO: 11, 34 or 71 and having any of the modifications discussed above, especially those set out in #l-#4 in the above paragraph.
  • the oligonucleotide preferably comprises or consists of the sequence shown in SEQ ID NO: 34 or 71 and having any of the modifications discussed above, especially those set out in #l-#4 in the above paragraph.
  • the oligonucleotide preferably comprises or consists of the sequence shown in any one of SEQ ID NOs: 29-32, 51-54, and 75-78.
  • the oligonucleotide preferably comprises or consists of the sequence shown in SEQ ID NO: 7, 8, 12, 18, 21, 39, 40, 41, 42 or 48 and having any of the modifications discussed above, especially those set out in #l-#4 in the above paragraph.
  • the oligonucleotide preferably comprises or consists of the sequence shown in SEQ ID NO: 7, 8, 12, 18, 21, 39, 40, 41, 42 or 48.
  • the oligonucleotide preferably comprises or consists of the sequence shown in SEQ ID NO: 7, 39 or 42 and having any of the modifications discussed above, especially those set out in #l-#4 in the above paragraph.
  • the oligonucleotide preferably comprises or consists of the sequence shown in SEQ ID NO: 7, 39 or 42.
  • One or more nucleotides in the oligonucleotide may be modified, for instance with a label or a tag.
  • the label may be any suitable label which allows the polynucleotide to be detected. Suitable labels include, but are not limited to, fluorescent molecules, radioisotopes, e.g., 125 1, 35 S, enzymes, antibodies, antigens, other polynucleotides, and ligands such as biotin.
  • the oligonucleotide may also be modified with a conjugate group.
  • Conjugate groups typically modify one or more properties of the oligonucleotide including, but not limited to, its pharmacodynamics, pharmacokinetics, stability, binding, absorption, cellular distribution, cellular uptake, charge, and clearance. Conjugate groups are routinely used in the art.
  • Conjugate groups include, but are not limited to, intercalators, reporter molecules, polyamines, polyamides, polyethylene glycols, thioethers, polyethers, cholesterols, thiocholesterols, cholic acid moieties, folate, lipids, phospholipids, biotin, phenazine, phenanthridine, anthraquinone, adamantane, acridine, fluoresceins, rhodamines, coumarins and dyes.
  • Certain conjugate groups have been described previously, including an N- Acetylgalactosamine (GalNAc) moiety and acholesterol moiety (Letsinger et al., Proc. Natl.
  • Acids Res., 1990, 18, 3777- 3783 a polyamine or a polyethylene glycol chain (Manoharan et al., Nucleosides & Nucleotides, 1995, 14, 969-973), or adamantane acetic acid (Manoharan et al., Tetrahedron Lett., 1995, 36, 3651-3654), a palmityl moiety (Mishra et al., Biochim. Biophys. Acta, 1995, 1264, 229-237), or an octadecylamine or hexylamino-carbonyl-oxycholesterol moiety (Crooke et al., J. Pharmacol. Exp. Then, 1996, 277, 923-937).
  • the conjugate group may comprise an active drug substance, such as aspirin, warfarin, phenylbutazone, ibuprofen, suprofen, fen-bufen, ketoprofen, (S)-(+)-pranoprofen, carprofen, dansylsarcosine, 2,3,5-triiodobenzoic acid, flufenamic acid, folinic acid, a benzothiadiazide, chlorothiazide, a diazepine, indo-methicin, a barbiturate, a cephalosporin, a sulfa drug, an antidiabetic, an antibacterial or an antibiotic.
  • an active drug substance such as aspirin, warfarin, phenylbutazone, ibuprofen, suprofen, fen-bufen, ketoprofen, (S)-(+)-pranoprofen, carprofen, dansylsarc
  • the label, tag or conjugate group may be directly attached to the oligonucleotide.
  • the label, tag or conjugate group may be attached to the oligonucleotide by a linking group.
  • Suitable linking groups include, but are not limited to, bifunctional linking moieties such as those known in the art.
  • conjugate linking moieties include pyrrolidine, 8-amino-3,6-dioxaoctanoic acid (ADO), succinimidyl 4-(N-maleimidomethyl) cyclohexa ne-1 -carboxylate (SMCC) and 6-aminohexanoic acid (AHEX or AHA).
  • linking groups include ethylene glycol-based linkers, for instance ethylene glycol based linkers with from about 2 to about 48 units and/or a molecular weight up to 5,000, and N- acetylgalactosamine (GalNAc) polymers.
  • ethylene glycol-based linkers for instance ethylene glycol based linkers with from about 2 to about 48 units and/or a molecular weight up to 5,000
  • GalNAc N- acetylgalactosamine
  • the label, tag or conjugate group may be attached to either or both ends of the oligonucleotide and/or at any internal position.
  • the oligonucleotide is preferably single stranded.
  • the oligonucleotide can be double stranded.
  • Oligonucleotides may be manufactured using standard techniques. Custom oligonucleotides having specific sequences and modifications are commercially available from various suppliers (such as IDT, SynOligo, RiboBio, Wuxi Apptec, and Gene Tools).
  • the oligonucleotide may be any length as long as it specifically hybridises to a target portion of the 3' UTR of HNRNPD mRNA and blocks at least one inhibitory miRNA binding site. As explained below, inhibitory miRNA binding sites are typically at least 5 nucleotides in length (see also SEQ ID NO: 79 below).
  • the oligonucleotide also needs to comprise sufficient nucleotides that allow it to specifically hybridise to the 3' UTR of HNRNPD mRNA.
  • the oligonucleotide is preferably from about 8 to about 50 nucleotide in length.
  • the oligonucleotide is preferably about 8, about 9, about 10, about 11, about 12, about 13, about 14, about 15, about 16, about 17, about 18, about 19, about 20, about 21, about 22, about 23, about 24, about 25, about 26, about 27, about 28, about 29, about 30, about 31, about 32, about 33, about 34, about 35, about 36, about 37, about 38, about 39, about 40, about 41, about 42, about 43, about 44, about 45, about 46, about 47, about 48, about 49, or about 50 nucleotides in length.
  • the oligonucleotide is preferably at least about 16 nucleotides in length.
  • the oligonucleotide is preferably at least about 16, about 17, about 18, about 19, about 20, about 21, about 22, about 23, about 24, about 25, about 26, about 27, about 28, about 29, about 30, about 31, about 32, about 33, about 34, about 35, about 36, about 37, about 38, about 39, about 40, about 41, about 42, about 43, about 44, about 45, about 46, about 47, about 48, about 49, or about 50 nucleotides in length.
  • the oligonucleotide is preferably from about 22 to about 27 nucleotides in length.
  • the oligonucleotide is preferably about 22, about 23, about 24, about 25, about 26, or about 27 nucleotides in length.
  • the oligonucleotide is most preferably about 25 nucleotides in length.
  • the oligonucleotide specifically hybridises to the target portion of the 3' UTR of HNRNPD mRNA.
  • the oligonucleotide "specifically hybridises" to the target portion when it hybridises with preferential or high affinity to the partner but does not substantially hybridise, does not hybridise, or hybridises with only low affinity to other polynucleotide sequences, especially other RNA sequences in the cell. This can be measured using techniques known in the art.
  • Hybridisation can be carried out under low stringency conditions, for example in the presence of a buffered solution of 30 to 35% formamide, 1 M NaCI and 1 % SDS (sodium dodecyl sulfate) at 37 °C followed by a 20-minute wash in from IX (0.1650 M Na+) to 2X (0.33 M Na+) SSC (standard sodium citrate) at 50 °C.
  • Hybridisation can be carried out under moderate stringency conditions, for example in the presence of a buffer solution of 40 to 45% formamide, 1 M NaCI, and 1 % SDS at 37 °C, followed by a wash in from 0.5X (0.0825 M Na+) to IX (0.1650 M Na+) SSC at 55 °C.
  • Hybridisation can be carried out under high stringency conditions, for example in the presence of a buffered solution of 50% formamide, 1 M NaCI, 1% SDS at 37 °C, followed by a wash in 0.1X (0.0165 M Na+) SSC at 60 °C.
  • the oligonucleotide "specifically hybridises” if it hybridises to the target portion with a melting temperature (Tm) that is at least 2 °C, such as at least 3 °C, at least 4 °C, at least 5 °C, at least 6 °C, at least 7 °C, at least 8 °C, at least 9 °C or at least 10 °C, greater than its Tm for other RNA sequences.
  • Tm melting temperature
  • the oligonucleotide hybridises to the target portion with a Tm that is at least 2 °C, such as at least 3 °C, at least 4 °C, at least 5 °C, at least 6 °C, at least 7 °C, at least 8 °C, at least 9 °C, at least 10 °C, at least 20 °C, at least 30 °C or at least 40 °C, greater than its Tm for other RNA sequences.
  • the oligonucleotide is preferably at least about 90% identical or homologous to the reverse complement of the target portion over its entire length.
  • the oligonucleotide is preferably at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical or homologous to the target portion over its entire length.
  • Homology or sequence identity is typically measured over the entire length of reference sequence (e.g., the target portion). This may also be referred to as global homology or global sequence identity.
  • the oligonucleotide may also have a lower sequence homology or identity to the reverse complement of the target portion because it comprises universal nucleotides which base pair with any nucleotide in the target portion.
  • a sequence having at least about 90% identity or homology to the reverse complement of the oligonucleotide is preferably found/present in the 3' UTR of HNRNPD mRNA but is not found in any other RNA molecule in the cell.
  • a sequence having at least about 90% identity or homology to the reverse complement of the oligonucleotide is preferably found/present in in the 3' UTR of HNRNPD mRNA but is not found in any other human RNA molecule.
  • the reverse complement of the oligonucleotide is preferably found/present in the 3' UTR of HNRNPD mRNA but is not found in any other RNA molecule in the cell.
  • the reverse complement of the oligonucleotide is preferably found/present in the 3' UTR of HNRNPD mRNA but is not found in any other human RNA molecule.
  • the UWGCG Package provides the BESTFIT program which can be used to calculate homology or identity (e.g., used on its default settings) (Devereux et al (1984) Nucleic Acids Research 12, p387-395).
  • the PILEUP and BLAST algorithms can also be used to calculate identity, homology, or line up sequences (typically on their default settings), for example as described in Altschul S.F. (1993) J Mol Evol 36:290-300; Altschul, S, F et al (1990) J Mol Biol 215:403-10.
  • HSPs high scoring sequence pair
  • Extensions for the word hits in each direction are halted when: the cumulative alignment score goes to zero or below, due to the accumulation of one or more negative-scoring residue alignments; or the end of either sequence is reached.
  • the BLAST algorithm parameters W, T and X determine the sensitivity and speed of the alignment.
  • the BLAST algorithm performs a statistical analysis of the similarity between two sequences; see e.g., Karlin and Altschul (1993) Proc. Natl. Acad. Sci. USA 90:5873-5787.
  • One measure of similarity provided by the BLAST algorithm is the smallest sum probability (P(N)), which provides an indication of the probability by which a match between two nucleotide or amino acid sequences would occur by chance.
  • P(N) the smallest sum probability
  • a sequence is considered similar to another sequence if the smallest sum probability in comparison of the first sequence to the second sequence is less than about 1, preferably less than about 0.1, more preferably less than about 0.01, and most preferably less than about 0.001.
  • the oligonucleotide typically comprises or consists of a sequence which is identical to the reverse complement of the target portion over at least about 8, at least about 9, at least about 10, at least about 11, at least about 12, at least about 13, at least about 14, at least about 15, at least about 16, at least about 17, at least about 18, at least about 19, at least about 20, at least about 21, at least about 22, at least about 23, at least about 24, at least about 25, at least about 26, at least about 27, at least about 28, at least about 29, at least about 30, at least about 31, at least about 32, at least about 33, at least about 34, at least about 35, at least about 36, at least about 37, at least about 38, at least about 39, at least about 40, at least about 41, at least about 42, at least about 43, at least about 44, at least about 45, at least about 46, at least about 47, at least about 48, or at least about 49 consecutive nucleotides.
  • the oligonucleotide typically comprises or consists of a sequence which is identical to the reverse complement of the target portion over at least about 12, at least about 13, at least about 14, at least about 15, at least about 16, at least about 17, at least about 18, at least about 19, at least about 20, at least about 21, at least about 22, at least about 23, at least about 24, at least about 25, at least about 26, or at least about 27 consecutive nucleotides.
  • the oligonucleotide most preferably comprises or consists of a sequence which is the reverse complement of the target portion. Complementarity based on Watson-Crick base pairing is well known in the art.
  • the target portion is typically the same length as the oligonucleotide.
  • the target portion is preferably from about 8 to about 50 nucleotide in length.
  • the target portion is preferably about 8, about 9, about 10, about 11, about 12, about 13, about 14, about 15, about 16, about 17, about 18, about 19, about 20, about 21, about 22, about 23, about 24, about 25, about 26, about 27, about 28, about 29, about 30, about 31, about 32, about 33, about 34, about 35, about 36, about 37, about 38, about 39, about 40, about 41, about 42, about 43, about 44, about 45, about 46, about 47, about 48, about 49, or about 50 nucleotides in length.
  • the target portion is preferably a portion of the sequence shown in SEQ ID NO: 79 as discussed below.
  • the target portion is preferably at least about 90%, such as at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99%, identical or homologous to the reverse complement of the sequence shown in any one of SEQ ID NOs: 1-28, 33-50, and 55-74.
  • the target portion is at least about 90%, such as at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99%, identical or homologous to the reverse complement of the sequence shown in SEQ ID NO: 11, 34 or 71.
  • the target portion is at least about 90%, such as at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99%, identical or homologous to the reverse complement of the sequence shown in SEQ ID NO: 34 or 71.
  • the oligonucleotide preferably blocks two or more inhibitory miRNA binding sites, such as 2, 3, 4 or more inhibitory miRNA binding sites.
  • the two or more inhibitory miRNA binding sites may be distinct. This means the two or more inhibitory miRNA binding sites are bound by different parts of the miRNA. An example of this are the two inhibitory binding sites shown in SEQ ID NOs: 81 and 82.
  • the two or more inhibitory miRNA binding sites may be overlapping. This means the two or more inhibitory miRNA binding sites are bound by overlapping parts of the miRNA. An example of this is shown in SEQ ID NOs: 84 and 85.
  • the two or more miRNA binding sites may have the same length.
  • the two or more inhibitory miRNA binding sites may have different lengths.
  • SOL044(0) blocks two inhibitory miRNA binding sites shown in SEQ ID NOs: 81 and 82, which have different lengths (5 nucleotides and 8 nucleotides respectively).
  • the oligonucleotide preferably blocks at least one miR-141-3p binding site.
  • the oligonucleotide preferably blocks two or more miR-141-3p binding sites.
  • the oligonucleotide preferably blocks at least one miR-146-5p binding site.
  • the oligonucleotide preferably blocks two or more miR-146-5p binding sites.
  • PFRED e.g., Sciabola S, Xi H, Cruz D, Cao Q, Lawrence C, Zhang T, Rotstein S, Hughes JD, Caffrey DR, Stanton RV.
  • PFRED A computational platform for siRNA and antisense oligonucleotides design.
  • PLoS One 2021 Jan 22;16(l):e0238753. doi: 10.1371/journal.pone.0238753.
  • PMID 33481821; PMCID: PMC7822268) and LNCASO (https://iomics.ugent.be/lncaso).
  • the at least one inhibitory miRNA binding site is typically at least 5 nucleotides in length.
  • the at least one inhibitory miRNA binding site is preferably from about 5 to about 10 nucleotides in length.
  • the at least one inhibitory miRNA binding site is preferably about 5, about 6, about 7 about 8, about 9, or about 10 nucleotides in length.
  • the at least one inhibitory miRNA binding site is preferably about 5, about 6 or about 8 nucleotides in length. These lengths equally apply to the two or more inhibitory miRNA binding sites.
  • the at least one inhibitory miRNA binding site typically comprises a stretch of consecutive nucleotides from the reverse complement of the inhibitory miRNA sequence.
  • the at least one inhibitory miRNA binding site typically comprises from about 5 to about 10 consecutive nucleotides, such as about 5, about 6, about 7, about 8, about 9 or about 10 consecutive nucleotides from the reverse complement of the inhibitory miRNA sequence.
  • the at least one inhibitory miRNA binding site typically comprises from about 5 to about 10 consecutive nucleotides, such as about 5, about 6, about 7, about 8, about 9 or about 10 consecutive nucleotides from the reverse complement of miR-141-3p or miR-146-5p.
  • SOL045(0) comprises the miR-146-5p binding site shown in SEQ ID NO: 84, which is 5 nucleotides in length.
  • SOL046(0) comprises the miR-146-5p binding site shown in SEQ ID NO: 85, which is 8 nucleotides in length.
  • the consecutive nucleotides are preferably from the about 10 nucleotides at the 3' end of the reverse complement of the inhibitory miRNA sequence, miR-141-3p or miR-146-5p.
  • the consecutive nucleotides are preferably from the 7 nucleotides at positions 2-8 from the 3' end of the reverse complement of the inhibitory miRNA sequence, miR-141-3p or miR-146- 5p. This is also known as the seed sequence or the seed region.
  • the oligonucleotide preferably comprises or consists of a sequence which is the reverse complement of the target portion and which comprises a sequence which is identical to at least about 5 consecutive nucleotides, such as at least about 5, about 6, about 7 about 8, about 9, or about 10 consecutive nucleotides, from the inhibitory miRNA sequence, miR- 141-3p or miR-146-5p.
  • the consecutive nucleotides are preferably from the about 10 nucleotides at the 5' end of the inhibitory miRNA sequence, miR-141-3p or miR-146-5p.
  • the consecutive nucleotides are preferably from the 7 nucleotides at positions 2-8 from the 5' end of inhibitory miRNA sequence, miR-141-3p or miR-146-5p. This is also known as the seed sequence or the seed region.
  • the oligonucleotide preferably comprises or consists of a sequence which is the reverse complement of the target portion and which comprises a sequence which is the reverse complement of the inhibitory miRNA binding site.
  • the target portion may have any of the lengths discussed above.
  • the miRNA binding site may be any of those discussed above.
  • the oligonucleotide preferably comprises or consists of a sequence which is the reverse complement of the target portion and which comprises a sequence which is identical to at least about 5 consecutive nucleotides, such as at least about 5, about 6, about 7 about 8, about 9, or about 10 consecutive nucleotides, from the sequence shown in SEQ ID NO: 80 or 83.
  • the target portion may have any of the lengths discussed above.
  • the consecutive nucleotides are preferably from the about 10 nucleotides at the 5' end of the sequence shown in SEQ ID NO: 80 or 83.
  • the consecutive nucleotides are preferably from the 7 nucleotides at positions 2-8 from the 5' end of the sequence shown in SEQ ID NO: 80 or 83. This is also known as the seed sequence or the seed region.
  • the oligonucleotide may comprise T in place of U in SEQ ID NO: 80 or 83.
  • the oligonucleotide preferably comprises or consists of a sequence which is the reverse complement of the target portion and which comprises the sequence(s) shown in SEQ ID NO: 81 and/or 82.
  • the target portion may have any of the lengths discussed above.
  • the oligonucleotide preferably comprises or consists of a sequence which is the reverse complement of the target portion and which comprises the sequence shown in SEQ ID NO: 84 or 85.
  • the target portion may have any of the lengths discussed above.
  • the sequences of the 3' UTRs of HNRNPD in various species are known in the art.
  • There are several isoforms of human HNRNPD including ENSG00000138668.20 and all of its associated isoforms, including ENST00000313899.12 (p45), ENST00000352301.8 (p42), ENST00000353341.8 (p40) and ENST00000503822.2 (p37)).
  • the invention may involve any of these isoforms.
  • the main HNRNPD isoforms are p45, p42, p40 and p37.
  • the sequence of the 3' UTR of human HNRNPD mRNA is the same in all four isoforms and is shown in SEQ ID NO: 79.
  • the 3' UTR preferably comprises or consists of the sequence shown in SEQ ID NO: 79.
  • the oligonucleotide preferably comprises or consists of a sequence which is the reverse complement of from about 8 to about 50 (or any of the lengths of the target portion set out above) nucleotides of the sequence shown in SEQ ID NO: 79 and which comprises a sequence which is identical to at least about 5 consecutive nucleotides, such as at least about 5, about 6, about 7 about 8, about 9, or about 10 consecutive nucleotides, from the inhibitory miRNA sequence, miR-141-3p or miR-146-5p.
  • the oligonucleotide preferably comprises or consists of a sequence which is the reverse complement of from about 8 to about 50 (or any of the lengths of the target portion set out above) nucleotides of the sequence shown in SEQ ID NO: 79 and which comprises a sequence which is the reverse complement to at least about 5 consecutive nucleotides, such as at least about 5, about 6, about 7 about 8, about 9, or about 10 consecutive nucleotides, from the inhibitory miRNA binding site.
  • the consecutive nucleotides are preferably from the about 10 nucleotides at the 5' end of the inhibitory miRNA sequence, miR-141-3p or miR-146-5p.
  • the consecutive nucleotides are preferably from the 7 nucleotides at positions 2-8 from the 5' end the inhibitory miRNA sequence, miR-141-3p or miR-146-5p. This is also known as the seed sequence or the seed region.
  • the nucleotides of the sequence shown in SEQ ID NO: 79 are preferably consecutive nucleotides from the sequence shown in SEQ ID NO: 79.
  • the oligonucleotide preferably comprises or consists of a sequence which is the reverse complement of from about 8 to about 50 nucleotides (or any of the lengths of the target portion set out above) of the sequence shown in SEQ ID NO: 79 and which comprises a sequence which is identical to at least about 5 consecutive nucleotides, such as at least about 5, about 6, about 7 about 8, about 9, or about 10 consecutive nucleotides, from the sequence shown in SEQ ID NO: 80 or 83.
  • the consecutive nucleotides are preferably from the about 10 nucleotides at the 5' end of the sequence shown in SEQ ID NO: 80 or 83.
  • the consecutive nucleotides are preferably from the 7 nucleotides at positions 2-8 from the 5' end of the sequence shown in SEQ ID NO: 80 or 83. This is also known as the seed sequence or the seed region.
  • the nucleotides of the sequence shown in SEQ ID NO: 79 are preferably consecutive nucleotides from the sequence shown in SEQ ID NO: 79.
  • the oligonucleotide may comprise T in place of U in SEQ ID NO: 80 or 83.
  • the oligonucleotide preferably comprises or consists of a sequence which is the reverse complement of from about 8 to about 50 (or any of the lengths of the target portion set out above) nucleotides of the sequence shown in SEQ ID NO: 79 and which comprises the sequence(s) shown in SEQ ID NO: 81 and/or 82.
  • the oligonucleotide preferably comprises or consists of a sequence which is the reverse complement of from about 8 to about 50 (or any of the lengths of the target portion set out above) nucleotides of the sequence shown in SEQ ID NO: 79 and which comprises the sequence shown in SEQ ID NO: 84 or 85.
  • the nucleotides of the sequence shown in SEQ ID NO: 79 are preferably consecutive nucleotides from the sequence shown in SEQ ID NO: 79.
  • the oligonucleotide preferably comprises or consists of the sequence shown in any one of SEQ ID NOs: 1-28, 33-50, and 55-74.
  • the oligonucleotide preferably comprises or consists of the sequence shown in any one of SEQ ID NOs: 1-26, 33-50, and 55-74.
  • the oligonucleotide preferably comprises or consists of the sequence shown in any one of SEQ ID NOs: 1-2, 5-9. 12, 13, 15-18, 20-26, 33-36, 38-50 and 55-74.
  • the oligonucleotide preferably comprises or consists of the sequence shown in SEQ ID NO: 11, 34 or 71.
  • the oligonucleotide preferably comprises or consists of the sequence shown in SEQ ID NO: 34 or 71. These oligonucleotides may comprise any of the modifications discussed above.
  • the oligonucleotide preferably comprises or consists of the sequence shown in any one of SEQ ID NOs: 29-32, 51-54, and 75-78.
  • the oligonucleotide preferably comprises or consists of the sequence shown in SEQ ID NO: 7, 8, 12, 18, 21, 39, 40, 41, 42 or 48.
  • the oligonucleotide preferably comprises or consists of the sequence shown in SEQ ID NO: 7, 39 or 42.
  • the cell or two or more cells is/are preferably in a subject.
  • the method preferably comprises administering the oligonucleotide or two or more oligonucleotides to the subject.
  • the invention therefore provides a method of reducing senescence in a cell or two or more cells in a subject, comprising administering to the subject an oligonucleotide or two or more oligonucleotides which specifically hybridise(s) to a target portion of the 3' untranslated region (UTR) of heterogeneous nuclear riboprotein particle D HNRNPD') mRNA and block(s) at least one inhibitory microRNA (miRNA) binding site and thereby reducing senescence in the cell or two or more cells.
  • UTR 3' untranslated region
  • D HNRNPD' heterogeneous nuclear riboprotein particle
  • miRNA inhibitory microRNA
  • the invention also provides an oligonucleotide or two or more oligonucleotides which specifically hybridise(s) to a target portion of the 3' UTR of HNRNPD mRNA and block(s) at least one inhibitory miRNA binding site for use in a method of reducing senescence in a cell or two or more cells in a subject.
  • the invention also provides use of an oligonucleotide or two or more oligonucleotides which specifically hybridise(s) to a target portion of the 3' UTR of HNRNPD mRNA and block(s) at least one inhibitory miRNA binding site in the manufacture of a medicament for reducing senescence in a cell or two or more cells in a subject.
  • an oligonucleotide comprising or consisting of the sequence shown in any one of SEQ ID NOs: 33-50 or in any of one of SEQ ID NOs: 33-36 and 38-50, or (c) an oligonucleotide comprising or consisting of the sequence shown in any one of SEQ ID NOs: 55-74, such as (a) and (b), (a) and (c), (b) and (c) and (a), (b) and (c).
  • the invention may comprise administering to the subject two or more of (a) an oligonucleotide comprising or consisting of the sequence shown in SEQ ID NO: 11, (b) an oligonucleotide comprising or consisting of the sequence shown in SEQ ID NO: 34, or (c) an oligonucleotide comprising or consisting of the sequence shown in SEQ ID NO: 72, such as (a) and (b), (a) and (c), (b) and (c) and (a), (b) and (c).
  • the invention may comprise administering to the subject an oligonucleotide comprising or consisting of the sequence shown in SEQ ID NO: 34 and/or an oligonucleotide comprising or consisting of the sequence shown in SEQ ID NO: 72.
  • the two or more oligonucleotides may be modified in any of the ways discussion above.
  • the invention may comprise administering to the subject two or more of (a) an oligonucleotide comprising or consisting of the sequence shown in SEQ ID NO: 7, (b) an oligonucleotide comprising or consisting of the sequence shown in SEQ ID NO: 39, or (c) an oligonucleotide comprising or consisting of the sequence shown in SEQ ID NO: 42, such as
  • oligonucleotides may be modified in any of the ways discussion above.
  • the subject is preferably mammalian.
  • the subject is preferably a human, dog, cat, primate, horse, murine, rat, rodent, bovine, murine, porcine, or ovine.
  • the subject is preferably a human.
  • the invention comprises administering to the subject an effective amount of the oligonucleotide or two or more oligonucleotides.
  • An effective amount is an amount which reduces senescence in the cell or two or more cells. Suitable amounts are discussed in more detail below.
  • the invention may be used in combination with other means of, and substances or compositions for, reducing senescence.
  • Such means, substances or compositions are known in the art.
  • the oligonucleotide(s) can be administered by one or more routes of administration using one or more of a variety of methods known in the art. As will be appreciated by the skilled person, the route and/or mode of administration will vary depending upon the desired results. Routes of administration include, but are not limited to, intravenous, intramuscular, intradermal, intraperitoneal, intrapleural, subcutaneous, spinal, or other parenteral routes of administration, for example by injection or infusion.
  • parenteral administration means modes of administration other than enteral and topical administration, usually by injection, and includes, without limitation, intravenous, intramuscular, intraarterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcuticular, intraarticular, subcapsular, subarachnoid, intraspinal, epidural, intrapleural and intra-sternal injection and infusion. Administration may be intrapleural or intraperitoneal.
  • the invention provides a method for treating or preventing (i) a lung disease or condition, (ii) an ILD associated with PPF or (ii) IPF in a subject, the method comprising administering to the subject by inhalation an oligonucleotide or two or more oligonucleotides which specifically hybridise(s) to a target portion of the 3' untranslated region (UTR) of heterogeneous nuclear riboprotein particle D HNRNPD) mRNA and block(s) at least one inhibitory microRNA (miRNA) binding site and thereby reducing senescence in one or more cells associated with the disease or condition, the ILD associated with PPF or the IPF.
  • UTR 3' untranslated region
  • HNRNPD heterogeneous nuclear riboprotein particle
  • miRNA inhibitory microRNA
  • the oligonucleotide(s) can be administered by a nonparenteral route, such as a topical, epidermal, or mucosal route of administration, for example, intranasally, orally, vaginally, rectally, sublingually, or topically.
  • a nonparenteral route such as a topical, epidermal, or mucosal route of administration, for example, intranasally, orally, vaginally, rectally, sublingually, or topically.
  • the oligonucleotide(s) may be for subcutaneous administration.
  • these carriers and/or diluents include aqueous or alcoholic/aqueous solutions, emulsions, or suspensions, including saline and/or buffered media.
  • Pharmaceutical compositions may be prepared together with a physiologically acceptable carrier or diluent.
  • the oligonucleotide(s) may be mixed with an excipient which is pharmaceutically acceptable and compatible with the active ingredient. Suitable excipients are, for example, water, saline, dextrose, glycerol, of the like and combinations thereof.
  • examples of pharmaceutically acceptable salts include, but are not limited to (a) salts formed with cations such as sodium, potassium, ammonium, magnesium, calcium, polyamines such as spermine and spermidine, etc.; (b) acid addition salts formed with inorganic acids, for example hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, nitric acid and the like; (c) salts formed with organic acids such as, for example, acetic acid, oxalic acid, tartaric acid, succinic acid, maleic acid, fumaric acid, gluconic acid, citric acid, malic acid, ascorbic acid, benzoic acid, tannic acid, palmitic acid, alginic acid, polyglutamic acid, naphthalenesulfonic acid, methanesulfonic acid, p- toluenesulfonic acid, naphthalenedisulfonic acid, polygalacturonic acid
  • Suspensions and emulsions may contain as carrier, for example a natural gum, agar, sodium alginate, pectin, methylcellulose, carboxymethylcellulose, or polyvinyl alcohol.
  • the suspensions or solutions for intramuscular injections may contain, together with the active substance, a pharmaceutically acceptable carrier, e.g., sterile water, olive oil, ethyl oleate, glycols, e.g., propylene glycol, and if desired, a suitable amount of lidocaine hydrochloride.
  • the pharmaceutical compositions are usually in the form of a sterile, pyrogen-free, parenterally acceptable composition.
  • a particularly suitable vehicle for parenteral injection is a sterile, isotonic solution, properly preserved.
  • the oligonucleotide(s) can be in the form of a lyophilizate, such as a lyophilized cake.
  • the pharmaceutical composition is preferably in a form suitable for inhalation, such as a solution, a suspension or a dry powder.
  • the pharmaceutical composition may comprise suitable carriers and/or excipients as discussed above.
  • the pharmaceutical composition may be in the form of lyophilised powder and may be administered as a dry powder.
  • compositions for topical administration may include transdermal patches, ointments, lotions, creams, gels, hydrogels, drops, suppositories, sprays, liquids, and powders.
  • Conventional pharmaceutical carriers, aqueous, powder or oily bases, thickeners and the like may be necessary or desirable.
  • compositions for subcutaneous administration contain suitable stabilizers (e.g., amino acids, such as methionine, and or saccharides such as sucrose), buffering agents and tonicifying agents.
  • suitable stabilizers e.g., amino acids, such as methionine, and or saccharides such as sucrose
  • binders and carriers may include, for example, polyalkylene glycols or triglycerides; such suppositories may be formed from mixtures containing the active ingredient in the range of 0.5% to 10%, preferably 1% to 2%.
  • compositions for oral administration may be syrups, emulsions, or suspensions.
  • the syrups may contain as carriers, for example, saccharose or saccharose with glycerine and/or mannitol and/or sorbitol.
  • Oral pharmaceutical compositions include such normally employed excipients as, for example, pharmaceutical grades of mannitol, lactose, starch, magnesium stearate, sodium saccharine, cellulose, magnesium carbonate, and the like. These pharmaceutical compositions typically take the form of solutions or suspensions and contain 10% to 95% of active ingredient, preferably 25% to 70%.
  • the lyophilised material may be reconstituted prior to administration, e.g., a suspension. Reconstitution is preferably effected in buffer.
  • the pharmaceutical composition administered may contain minor amounts of auxiliary substances such as wetting or emulsifying agents, pH buffering agents, and/or adjuvants which enhance effectiveness.
  • the pharmaceutical composition preferably comprises human serum albumin.
  • Plasma-Lyte A® is a sterile, nonpyrogenic isotonic solution for intravenous administration.
  • Each 100 mL contains 526 mg of Sodium Chloride, USP (NaCI); 502 mg of Sodium Gluconate (C6HllNaO7); 368 mg of Sodium Acetate Trihydrate, USP (C2H3NaO2*3H2O); 37 mg of Potassium Chloride, USP (KCI); and 30 mg of Magnesium Chloride, USP (MgCI2*6H2O). It contains no antimicrobial agents.
  • the pH is adjusted with sodium hydroxide. The pH is 7.4 (6.5 to 8.0).
  • Suitable further agents for inclusion in the pharmaceutical composition include, but are not limited to, amino acids (such as glycine, glutamine, asparagine, arginine, or lysine), antimicrobials, antioxidants (such as ascorbic acid, sodium sulphite, or sodium hydrogensulphite), buffers (such as borate, bicarbonate, Tris-HCI, citrates, phosphates, or other organic acids), bulking agents (such as mannitol or glycine), chelating agents (such as ethylenediamine tetraacetic acid (EDTA)), complexing agents (such as caffeine, polyvinylpyrrolidone, beta-cyclodextrin, or hydroxypropyl-beta-cyclodextrin), fillers, monosaccharides, disaccharides, and other carbohydrates (such as glucose, mannose, or dextrins), proteins (such as free serum albumin, gelatin, or immunoglobulins), colouring, flavouring and diluti
  • the oligonucleotide(s) may be formulated as a dermatological composition.
  • the pharmaceutical composition of the invention may be a dermatological composition. Such compositions may be used to treat or prevent one or more signs of ageing in a subject.
  • the dermatological composition is preferably administered by gymnosis.
  • the dermatological composition may comprise any of the pharmaceutical carriers and/or diluents discussed above.
  • the dermatological composition may comprise any of the pharmaceutical carriers and/or diluents discussed above with reference to topical compositions.
  • the dermatological composition may have the appearance of a white or coloured cream, an ointment, a milk, a lotion, a serum, a paste or of a foam.
  • the dermatological composition can be an aerosol.
  • the dermatological composition may also be in pulverulent solid form, for example in stick form.
  • the dermatological composition may be in the form of a patch, a pencil, a brush, or an applicator. These allow localized application to areas of the skin, hair, or nails.
  • the dermatological composition can also be formulated as a care product and/or as a makeup product.
  • the dermatological composition may be an emulsion.
  • the proportion of the fatty phase may range from about 5% to about 80% by weight, and preferably from about 5% to 50% by weight relative to the total weight of the composition.
  • the oils, emulsifiers and coemulsifiers used in the emulsion form are chosen from those conventionally used in the field.
  • the emulsifier and the coemulsifier are present in the composition in a proportion ranging from about 0.3% to about 30% by weight, and preferably from about 0.5% to about 20% by weight relative to the total weight of the composition.
  • Suitable oils for use in the dermatological composition include, but are not limited to, mineral oils, such as liquid petrolatum, vegetable oils, such as avocado oil or soybean oil, oils of animal origin, such as lanolin, synthetic oils, such as perhydrosqualene, silicone oils, such as cyclomethicone, and fluorinated oil,s such as perfluoropolyethers. It is also possible to use fatty alcohols, such as cetyl alcohol, fatty acids or waxes, such as carnauba wax, or ozokerite.
  • emulsifiers and coemulsifiers examples include, for example, fatty acid and polyethylene glycol esters, such as PEG-20 stearate, and fatty acid and glycerol esters, such as glyceryl stearate.
  • the dermatological composition may comprise a hydrophilic gelling agent, such as carboxyvinyl polymers (carbomer), acrylic copolymers such as copolymers of acrylates/alkyl acrylates, polyacrylamides, polysaccharides, gums natural and clays.
  • a hydrophilic gelling agent such as carboxyvinyl polymers (carbomer), acrylic copolymers such as copolymers of acrylates/alkyl acrylates, polyacrylamides, polysaccharides, gums natural and clays.
  • the dermatological composition may comprise a lipophilic gelling agent, such as a modified clay, such as a bentone, a metal salt of fatty acids, a hydrophobic silica or a polyethylene.
  • the oligonucleotide(s) may be administered by gymnosis. In other words, the oligonucleotide(s) may be administered without any transfection reagent.
  • the oligonucleotide(s) may be administered with a transfection reagent.
  • the pharmaceutical composition or dermatological composition may comprise a transfection reagent.
  • the transfection reagent may be liposomes, preferably cationic liposomes, exosomes, polymers, preferably cationic polymers, and dendrimers.
  • Other transfection reagents include, but are not limited to, macromolecule complexes, nanocapsules, microspheres, beads and lipid- based systems including oil-in-water emulsions, micelles, mixed micelles, and lipid :oligonucleotide complexes of uncharacterized structure.
  • Liposomes are preferred transfection reagents. Liposomes are typically microscopic spheres having an aqueous core surrounded by one or more outer layers made up of lipids arranged in a bilayer configuration (see, generally, Chonn et al., Current Op. Biotech. 1995, 6, 698- 708).
  • the liposomes that may be used in the invention include, but are not limited to, multilamellar liposomes or MLVs (MultiLamellar Vesicle), small unilamellar liposomes or SUVs (Small Unilamellar Vesicle), or large unilamellar liposomes or LUVs (Large Unilamellar Vesicle).
  • the liposomes may also be a nonionic liposomes whose wall is composed nonionic lipids.
  • the transfection agents may be lipfectamine® or oligofectamine®.
  • the transfection agent is preferably a pharmaceutically acceptable transfection agent. Such agents are suitable for administration to subjects and delivery of oligonucleotides.
  • the pharmaceutically acceptable transfection reagent is preferably a pharmaceutically acceptable PEI transfection reagent, such as a linear PEI transfection reagent.
  • the pharmaceutical acceptable transfection reagent may be GMP in vivo-jetPEI®.
  • fatty acids and their derivatives which act as penetration enhancers include, for example, oleic acid, lauric acid, capric acid, myristic acid, palmitic acid, stearic acid, linoleic acid, linolenic acid, dicaprate, tricaprate, recinleate, monoolein (a.k.a.
  • Chelating agents include, but are not limited to, disodium ethylenediaminetetraacetate (EDTA), citric acid, salicylates (e.g., sodium salicylate, 5-methoxysalicylate and homovanilate), N-acyl derivatives of collagen, laureth-9 and N-amino acyl derivatives of beta-diketones (enamines) [Lee et al., Critical Reviews in Therapeutic Drug Carrier Systems 1991, page 92; Muranishi, Critical Reviews in Therapeutic Drug Carrier
  • Chelating agents have the added advantage of also serving as DNase inhibitors.
  • Non-surfactants include, for example, unsaturated cyclic ureas, 1 -alkyl- and 1- alkenylazacyclo-alkanone derivatives (Lee et al., Critical Reviews in Therapeutic Drug Carrier Systems 1991, page 92); and non-steroidal anti-inflammatory agents such as diclofenac sodium, indomethacin and phenylbutazone (Yamashita et al., J. Pharm. Pharmacol. 1987, 39, 621-626).
  • oligonucleotide(s) may be administered in accordance with any of the strategies described in Roberts, T.C., Langer, R. & Wood, MJ. A. Advances in oligonucleotide drug delivery. Nat Rev Drug Discov 19, 673-694 (2020).
  • any suitable dose of the oligonucleotide(s) may be administered to a subject.
  • the dose may be determined according to various parameters, especially according to the oligonucleotide(s) used, the age, weight, and condition of the subject to be treated, the route of administration, and the required regimen. Again, a physician will be able to determine the required route of administration and dosage for any particular subject.
  • a typical daily dose is from about 0.01 to 50 mg per kg of body weight, according to the activity of the specific substance, the age, weight, and conditions of the subject to be treated and the frequency and route of administration.
  • daily dosage levels are from 5 mg to 2 g.
  • from about 0.01 to about 50mg per kg of subject of sRNA may administered, such as from about 0.05 to about 40, from about 0.1 to about 30, from about 0.5 to about 20, from about 1 to about 10 or from about 2 to about 5 mg per kg.
  • At least about 0.01 mg per kg of subject may administered, such as at least about 0.05, at least about 0.1, at least about 0.5, at least about 1, at least about 2, at least about 5, at least about 10, at least about 20, at least at least about 30 or at least about 40 mg per kg.
  • doses may be provided as a single dose or may be provided as multiple doses, for example taken at regular intervals, for example 2, 3 or 4 doses administered daily.
  • regular intervals include, but are not limited to, every day, every week, every fortnight, or every month.
  • the method of the invention is preferably for treating or preventing a disease or condition in the subject.
  • the invention therefore provides a method of treating or preventing a disease or condition in a subject, comprising administering to the subject an oligonucleotide or two or more oligonucleotides which specifically hybridise(s) to a target portion of the 3' untranslated region (UTR) of heterogeneous nuclear riboprotein particle D HNRNPD) mRNA and block(s) at least one inhibitory microRNA (miRNA) binding site and thereby reducing senescence in one or more cells associated with the disease or condition.
  • This method may involve any of the embodiments discussed above.
  • the invention also provides an oligonucleotide or two or more oligonucleotides which specifically hybridise(s) to a target portion of the 3' UTR of HNRNPD mRNA and block(s) at least one inhibitory miRNA binding site for use in a method of treating or preventing a disease or condition in a subject.
  • the invention also provides use of an oligonucleotide or two or more oligonucleotides which specifically hybridise(s) to a target portion of the 3' UTR of HNRNPD mRNA and block(s) at least one inhibitory miRNA binding site in the manufacture of a medicament for treating or preventing a disease or condition in a subject.
  • the oligonucleotide(s) may be any of those discussed above, including the modified oligonucleotides.
  • the oligonucleotide(s) may be administered in the form of a pharmaceutical composition as described above or a pharmaceutical composition of the invention.
  • oligonucleotides such as 3, 4, 5 or 6 oligonucleotides, as defined above or of the invention may be administered to the subject or may be used. Any of the embodiments discussed above are equally applicable to the therapeutic embodiments of the invention.
  • the subject is preferably mammalian.
  • the subject is preferably a human, dog, cat, primate, horse, murine, rat, rodent, bovine, murine, porcine, or ovine.
  • the subject is preferably a human.
  • the disease is preferably associated with senescence.
  • the disease or condition is preferably associated with increased senescence in one or more cells. This can be measured as described above.
  • One or more cells associated with the disease or condition in the subject preferably demonstrate increased senescence.
  • the skilled person is capable of identifying the one or more cells. For instance, the skilled person is capable of identifying the one or more lung fibroblasts associated with idiopathic pulmonary fibrosis (IPF).
  • IPF idiopathic pulmonary fibrosis
  • the oligonucleotide reduces senescence in the one or more cells associated with the disease or condition.
  • the disease or condition is preferably an interstitial lung disease (ILD).
  • the ILD may be Acute Chest Syndrome, Acute Respiratory Distress Syndrome (ARDS), Alpha-1 Antitrypsin Deficiency, Asbestosis, Aspergillosis, Asthma, Atelectasis, Bronchiectasis, Bronchiolitis, Bronchiolitis Obliterans (Popcorn Lung), Bronchitis (Acute), Bronchopulmonary Dysplasia, Chronic Bronchitis, Chronic Cough, Chronic Lung Disease, Chronic Thromboembolic Pulmonary Hypertension (CTEPH), Coal Worker's Pneumoconiosis (Black Lung Disease), COPD, Coccidioidomycosis (Valley Fever), Coronavirus, COVID-19, Common Cold, Cryptogenic Organizing Pneumonia (COP), Cystic Fibrosis (CF), E-cigarette or Vaping Use- Associated Lung Injury (EVALI), Emphysema
  • the disease or condition may be an interstitial lung disease (ILD) associated with progressive pulmonary fibrosis (PPF).
  • the ILD associated with PPF may be connective tissue disease (CTD), genetic and/or familial pulmonary fibrosis (G/F PF), hypersensitivity pneumonitis (HP), idiopathic non-specific interstitial pneumonia (iNSIP), interstitial pneumonia (IPAF) preferably with autoimmune features, idiopathic pulmonary fibrosis (IPF) or an unclassifiable ILD (uILD).
  • the disease or condition is preferably idiopathic pulmonary fibrosis (IPF).
  • the disease or condition is preferably selected from idiopathic pulmonary fibrosis (IPF), chronic obstructive pulmonary disease (COPD), renal fibrosis, chronic kidney disease, age related macular degeneration (AMD), osteoarthritis, osteoporosis, rheumatoid arthritis, nonalcoholic fatty liver disease, non-alcoholic steatohepatitis, chronic wounds, radiation wounds, blistering diseases, actinic keratosis, systemic sclerosis, non-segmental vitiligo, papular pruritic eruption, alopecia, senile pruritis, Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis (ALS), frontotemporal dementia, cardiovascular disease, hypertension, type 2 diabetes, cancer, cognitive dysfunction, frailty, progeroid syndromes, and any disease in which senescence is implicated.
  • IPF idiopathic pulmonary fibrosis
  • COPD chronic obstructive pulmonary
  • the disease or condition is preferably selected from idiopathic pulmonary fibrosis (IPF), radiation-induced dermatitis (RD), chronic obstructive pulmonary disease (COPD), age related macular degeneration (AMD), osteoarthritis, and osteoporosis.
  • IPF idiopathic pulmonary fibrosis
  • RD radiation-induced dermatitis
  • COPD chronic obstructive pulmonary disease
  • AMD age related macular degeneration
  • osteoarthritis osteoporosis
  • the method typically comprises administering a therapeutically effective amount or a prophylactically effective amount of the oligonucleotide(s).
  • a therapeutically effective amount is an amount which ameliorates one or more symptoms, such as all the symptoms, of the disease or condition and/or abolishes one or more symptoms, such as all the symptoms, of the disease or condition.
  • the therapeutically effective amount preferably cures the disease.
  • a prophylactically effective amount is an amount which prevents the onset of the disease or condition and/or prevents the onset of one or more symptoms, such as all the symptoms, of the disease or condition.
  • the prophylactically effective amount preferably prevents the subject from developing the disease. Suitable amounts are discussed in more detail above.
  • the oligonucleotide(s) or the pharmaceutical composition may be administered to a subject that displays symptoms of the disease or condition.
  • One or more cells associated with the disease or condition typically demonstrate increased senescence.
  • the oligonucleotide(s) or the pharmaceutical composition may be administered to a subject that is asymptomatic, i.e. does not display symptoms of disease. Despite the lack of symptoms, one or more cells associated with the disease or condition may demonstrate increased senescence.
  • the oligonucleotide(s) or the pharmaceutical composition may be administered when the subject's disease status is unknown or the patient is expected not to have a disease or condition.
  • the oligonucleotide(s) or the pharmaceutical composition may be administered to a subject that is predisposed, such as genetically predisposed, to developing the disease or condition.
  • the oligonucleotide(s) or the pharmaceutical composition may be administered to a refractory subject or a subject otherwise contraindicated to first line treatment.
  • the invention may be used in combination with other means of, and substances or compositions for, treating or preventing the disease or condition or providing pain relief.
  • the oligonucleotide(s) may be used in combination with existing treatments for the disease or condition including intensive care treatment. These include one or more of standard of care treatments, anti-inflammatories, anti-fibrotics and vasodilators.
  • intensive care treatment include one or more of standard of care treatments, anti-inflammatories, anti-fibrotics and vasodilators.
  • ILDs associated with PPF and IPF the invention may be combined with pirfenidone, nintedanib, or any of the drugs discussed in Bonella F, Spagnolo P, Ryerson C. Current and Future Treatment Landscape for Idiopathic Pulmonary Fibrosis. Drugs. 2023 Nov;83(17): 1581-1593. doi: 10.1007/s40265-023-01950-0.
  • the method of the invention is preferably for the treatment or prevention of one or more signs of ageing in the subject.
  • the invention therefore provides a method of treating or preventing one or more signs of ageing in a subject, comprising administering to the subject an oligonucleotide or two or more oligonucleotides which specifically hybridise(s) to a target portion of the 3' untranslated region (UTR) of heterogeneous nuclear riboprotein particle D HNRNPD) mRNA and block(s) at least one inhibitory microRNA (miRNA) binding site and thereby reducing senescence in one or more cells associated with the one or more signs of ageing.
  • the method is a non-therapeutic method. The method may reverse, delay or slow one or more signs of ageing in the subject.
  • the oligonucleotide(s) may be any of those discussed above, including the modified oligonucleotides.
  • the oligonucleotide(s) may be administered in the form of a dermatological composition as described above or a dermatological composition of the invention.
  • the one or more cells may be any of those discussed above.
  • the one or more cells are preferably one or more skin cells.
  • the one or more cells are preferably one or more skin fibroblasts.
  • the one or more cells are preferably one or more skin cells.
  • the one or more cells are preferably one or more hair follicle cells.
  • the one or more cells are preferably one or more hair follicle stem cells.
  • the one or more cells are preferably one or more nail cells.
  • the one or more cells are preferably one or more nail matrix cells.
  • the invention also provides an oligonucleotide or two or more oligonucleotide(s) which specifically hybridise(s) to a target portion of the 3' UTR of HNRNPD mRNA and block(s) at least one inhibitory miRNA binding site for use in a method of treating or preventing one or more signs of ageing in a subject.
  • the invention also provides use of an oligonucleotide or two or more oligonucleotide(s) which specifically hybridise(s) to a target portion of the 3' UTR of HNRNPD mRNA and block(s) at least one inhibitory miRNA binding site in the manufacture of a medicament for treating or preventing one or more signs of ageing in a subject.
  • the oligonucleotide(s) may be any of those discussed above, including the modified oligonucleotides.
  • the oligonucleotide(s) may be administered in the form of a dermatological composition as described above or a cosmetic or dermatological composition of the invention.
  • oligonucleotides such as 3, 4, 5 or 6 oligonucleotides, as defined above or of the invention may be administered to the subject or may be used. Any of the embodiments discussed above are equally applicable to the ageing embodiments of the invention.
  • the subject is preferably mammalian.
  • the subject is preferably a human, dog, cat, primate, horse, murine, rat, rodent, bovine, murine, porcine, or ovine.
  • the subject is preferably a human.
  • compositions and dosages are defined above. Any of these may be used in the ageing embodiments of the invention.
  • any number of signs of ageing may be treated or prevented in accordance with the invention, such as about 2, about 3, about 4, about 5, about 6, about 7, about 8, about 9, or about 10 signs of ageing.
  • the one or more signs of ageing in the hair may be selected from hair loss, hair thinning, hair becoming brittle, a reduced rate and/or length of hair growth, and a loss of hair pigmentation.
  • the oligonucleotide(s) or the dermatological composition may be administered to a subject that displays one or more signs of ageing.
  • One or more cells associated with the one or more signs of ageing typically demonstrate increased senescence.
  • the oligonucleotide(s) or the dermatological composition may be administered to a subject that does not display the one or more signs of ageing. Despite the lack of signs, one or more cells associated with the one or more signs of ageing may demonstrate increased senescence.
  • the oligonucleotide(s) or the dermatological composition may be administered when the subject's status is unknown or the patient is expected not to have the one or more signs of ageing.
  • the oligonucleotide(s) or the dermatological composition may be administered to a subject that is predisposed, such as genetically predisposed, to developing the one or more signs of ageing.
  • the invention may be used in combination with other means of, and substances or compositions for, treating or preventing the one or more signs of ageing.
  • the oligonucleotide(s) may be used in combination with existing anti-ageing treatments.
  • the invention also provides an oligonucleotide of about 50 or fewer nucleotides and comprising or consisting of any one of the sequences shown in SEQ ID NOs: 1-28, 33-50, and 55-74.
  • the oligonucleotide may have any length of about 50 or fewer nucleotides, including about 49 or fewer, about 48 or fewer, about 47 or fewer, about 46 or fewer, about 45 or fewer, about 44 or fewer, about 43 or fewer, about 42 or fewer, about 41 or fewer, about 40 or fewer, about 39 or fewer, about 38 or fewer, about 37 or fewer, about 36 or fewer, about 35 or fewer, about 34 or fewer, about 33 or fewer, about 32 or fewer, about 31 or fewer, about 30 or fewer, about 29 or fewer, about 28 or fewer, about 27 or fewer, or about 26 or fewer nucleotides.
  • the nucleotides may be any of those discussed above with reference to the methods
  • the oligonucleotide preferably comprises or consists of any one of the sequences shown in SEQ ID NOs: 1-26, 33-50, and 55-74.
  • the oligonucleotide preferably comprises or consists of any one of the sequences shown in SEQ ID NOs: 1-2, 5-9. 12, 13, 15-18, 20-26, 33-36, 38-50 and 55-74.
  • the oligonucleotide preferably comprises or consists of the sequence shown in SEQ ID NO: 11, 34 or 71.
  • the oligonucleotide preferably comprises or consists of the sequence shown in SEQ ID NO: 34 or 71.
  • the oligonucleotide preferably comprises or consists of the sequence shown in SEQ ID NO: 7, 8, 12, 18, 21, 39, 40, 41, 42 or 48.
  • the oligonucleotide preferably comprises or consists of the sequence shown in SEQ ID NO: 7, 39 or 42.
  • the oligonucleotide is preferably from about 22 to about 27 nucleotides in length, such as about 22, about 23, about 24, about 25, about 26 or about 27 nucleotides in length.
  • the oligonucleotide is preferably about 25 nucleotides in length.
  • the oligonucleotide preferably comprises one or more 2’-O-methoxyethyl (2' MOE) nucleotides and/or one or more 2'-O-methyl (OMe) nucleotides.
  • the oligonucleotide preferably comprises one or more one or more phosphorothioate (PS) internucleoside linkages and/or one or more phosphorodiamidate morpholino (PMO) internucleoside linkages.
  • PS phosphorothioate
  • PMO phosphorodiamidate morpholino
  • the oligonucleotide preferably comprises any of the preferred modifications discussed above with reference to the method of the invention, including those numbered #l-#4.
  • the oligonucleotide preferably comprises one of the following:
  • the oligonucleotide preferably comprises or consists of the sequence shown in SEQ ID NOs: 29-32, 51-54, and 75-78.
  • oligonucleotide of the invention may also be modified with a tag, label or conjugate group.
  • tag, label or conjugate group Such groups are described above with reference to the method of the invention and any of those embodiments equally apply to the oligonucleotides of the invention.
  • the oligonucleotide of the invention may be used in any of the methods described above.
  • the oligonucleotide of the invention may be used in therapy.
  • the oligonucleotide of the invention may be used as a prophylactic.
  • the oligonucleotide may be isolated, substantially isolated, purified or substantially purified.
  • the oligonucleotide is isolated or purified if it is completely free of any other components, such as buffer, other oligonucleotides, production material or cells.
  • the oligonucleotide is substantially isolated or substantially purified if it is only mixed with carriers or diluents, such as buffers or excipients, which will not interfere with its intended use. The oligonucleotide is not naturally occurring.
  • compositions of the invention are provided.
  • the invention also provides compositions for use in the invention.
  • the invention provides a pharmaceutical composition comprising an oligonucleotide of the invention and a pharmaceutically acceptable carrier and/or diluent. Suitable carriers and diluents are discussed above with reference to the method of the invention.
  • the pharmaceutical composition preferably comprises a pharmaceutically acceptable transfection agent. Any of the transfection agents discussed above may be used.
  • the transfection agent preferably comprises liposomes.
  • the pharmaceutical composition preferably comprises one or more penetration enhancers. Such enhancers are discussed above.
  • the pharmaceutical composition is preferably a dermatological composition.
  • the dermatological composition may comprise any of the dermatological acceptable carriers and/or diluents described above.
  • the dermatological composition may comprise an emulsion.
  • the pharmaceutical or dermatological composition may comprise two or more oligonucleotides of the invention, such as 3, 4, 5 or 6 oligonucleotides of the invention.
  • the pharmaceutical or dermatological composition may comprise two or more of (a) an oligonucleotide of the invention comprising or consisting of the sequence shown in any one of SEQ ID NOs: 1-28, in any one of SEQ ID NOs: 1-26, in any one of SEQ ID NOs: 1-2, 5-9.
  • an oligonucleotide of the invention comprising or consisting of the sequence shown in any one of SEQ ID NOs: 33-50 or in any of one of SEQ ID NOs: 33-36 and 38-50, or (c) an oligonucleotide of the invention comprising or consisting of the sequence shown in any one of SEQ ID NOs: 55-74, such as (a) and (b), (a) and (c), (b) and (c) and (a), (b) and (c).
  • the two or more oligonucleotides of the invention may be modified in any of the ways discussed above.
  • the pharmaceutical or dermatological composition may comprise two or more of (a) an oligonucleotide of the invention comprising or consisting of the sequence shown in SEQ ID NO: 11, (b) an oligonucleotide of the invention comprising or consisting of the sequence shown in SEQ ID NO: 34, or (c) an oligonucleotide of the invention comprising or consisting of the sequence shown in SEQ ID NO: 72, such as (a) and (b), (a) and (c), (b) and (c) and (a), (b) and (c).
  • the pharmaceutical or dermatological composition may comprise an oligonucleotide of the invention comprising or consisting of the sequence shown in SEQ ID NO: 34 and/or an oligonucleotide of the invention comprising or consisting of the sequence shown in SEQ ID NO: 72.
  • the pharmaceutical or dermatological composition may comprise two or more of (a) an oligonucleotide of the invention comprising or consisting of the sequence shown in SEQ ID NO: 7, (b) an oligonucleotide of the invention comprising or consisting of the sequence shown in SEQ ID NO: 39, or (c) an oligonucleotide of the invention comprising or consisting of the sequence shown in SEQ ID NO: 42, such as (a) and (b), (a) and (c), (b) and (c) and (a), (b) and (c).
  • the invention also provides an inhalation device, such as a metered dose inhaler (MDI), comprising a therapeutically effective amount or a prophylactically effective amount of an oligonucleotide of the invention or two or more oligonucleotides of the invention.
  • MDI metered dose inhaler
  • the invention also provides a cartridge adapted for use with an inhalation device or a MDI, comprising a therapeutically effective amount or a prophylactically effective amount of an oligonucleotide of the invention or two or more oligonucleotides of the invention.
  • the inhalation device, MDI or cartridge of the invention may be for treating any of the diseases or conditions discussed above, including a lung disease or condition, an ILD, an ILD associated with PPF or IPF.
  • mice were intratracheally administered with the therapeutic oligonucleotide once at Day 0, at a total volume of ⁇ 50 pl, corresponding to a final dose of lOmg/kg, under recoverable anaesthesia (isoflurane/oxygen mix).
  • sample lysate was then centrifuged at 18000 x g, 10 min, 4°C to pellet and remove debris.
  • MOE/DNA-PS oligonucleotide While alternating MOE/DNA-PS oligonucleotide was rapidly released from lungs after 24h, the morpholino modified oligonucleotide was still detectable after 168h post administration.
  • the MOE-PS oligonucleotide showed the most favourable PK profile, with a characteristic slow accumulation in the lungs up to 48h, followed by a drop at 72h and a second slow accumulation to 168h.

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Abstract

L'invention concerne de nouveaux oligonucléotides et leur utilisation pour inhiber ou réduire la sénescence. Les oligonucléotides ciblent des séquences spécifiques dans la région non traduite 3' (UTR) d'ARNm de la ribonucléoprotéine nucléaire hétérogène D (HNRNPD) et bloque un ou plusieurs sites de liaison de microARN inhibiteur (miARN). Ceci favorise l'expression physiologiquement régulée de HNRNPD et réduit la sénescence.
PCT/EP2025/058355 2024-03-26 2025-03-26 Nouveaux oligonucléotides Pending WO2025202340A2 (fr)

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GB2404290.5 2024-03-26
GBGB2404290.5A GB202404290D0 (en) 2024-03-26 2024-03-26 Novel oligoncleotides

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