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WO2018197926A1 - Procédés de création de nouveaux agents antibactériens à l'aide d'oligonucléotides antisens chimériques - Google Patents

Procédés de création de nouveaux agents antibactériens à l'aide d'oligonucléotides antisens chimériques Download PDF

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WO2018197926A1
WO2018197926A1 PCT/IB2017/052402 IB2017052402W WO2018197926A1 WO 2018197926 A1 WO2018197926 A1 WO 2018197926A1 IB 2017052402 W IB2017052402 W IB 2017052402W WO 2018197926 A1 WO2018197926 A1 WO 2018197926A1
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seq
bacterial growth
aso
inhibition
staphylococcus
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Robert Penchovsky
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    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/04—Antibacterial agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/62—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being a protein, peptide or polyamino acid
    • A61K47/64—Drug-peptide, drug-protein or drug-polyamino acid conjugates, i.e. the modifying agent being a peptide, protein or polyamino acid which is covalently bonded or complexed to a therapeutically active agent
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/62—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being a protein, peptide or polyamino acid
    • A61K47/64—Drug-peptide, drug-protein or drug-polyamino acid conjugates, i.e. the modifying agent being a peptide, protein or polyamino acid which is covalently bonded or complexed to a therapeutically active agent
    • A61K47/645—Polycationic or polyanionic oligopeptides, polypeptides or polyamino acids, e.g. polylysine, polyarginine, polyglutamic acid or peptide TAT
    • A61K47/6455—Polycationic oligopeptides, polypeptides or polyamino acids, e.g. for complexing nucleic acids
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    • 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
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    • C12N2310/00—Structure or type of the nucleic acid
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    • C12N2310/00—Structure or type of the nucleic acid
    • C12N2310/30—Chemical structure
    • C12N2310/31—Chemical structure of the backbone
    • C12N2310/318—Chemical structure of the backbone where the PO2 is completely replaced, e.g. MMI or formacetal
    • C12N2310/3181—Peptide nucleic acid, PNA
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    • C12N2310/30—Chemical structure
    • C12N2310/32—Chemical structure of the sugar
    • C12N2310/323—Chemical structure of the sugar modified ring structure
    • C12N2310/3231—Chemical structure of the sugar modified ring structure having an additional ring, e.g. LNA, ENA
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    • C12N2310/00—Structure or type of the nucleic acid
    • C12N2310/30—Chemical structure
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    • C12N2310/341—Gapmers, i.e. of the type ===---===
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    • C12N2310/30—Chemical structure
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    • C12N2310/00—Structure or type of the nucleic acid
    • C12N2310/30—Chemical structure
    • C12N2310/35—Nature of the modification
    • C12N2310/351—Conjugate
    • C12N2310/3513—Protein; Peptide

Definitions

  • FIELD OF THE INVENTION Development of novel antibacterial agents by the use of chimeric antisense oligonucleotides coupled with cell penetrating peptides that bind specific bacterial RNAs and inhibit their functions.
  • Antisense chimeric oligonucleotides-oligopeptide molecules The peptides start with their N-ends.
  • PNA stands for Peptide nucleic acid
  • PS-DNA stands for Phosphorothioate 225 DNA
  • LNA stands for Locked Nucleic Acid. All nucleic acid oligonucleotides start with their 5'-ends.
  • down case "i” stands for "2'-0-CH3" of the DNA oligomers while the down case “2” stands PS -modification of DNA the DNA oligomers.
  • the "aar” stands for amino acid residues while the "nt” stands for nucleotides.
  • mRNA PS-DNA G l A l T l T2T2T2G2C2T2T2C2T2T2T2A2 A2C2C2T2A2 A iT 1 T 1
  • CAACTTCTTTTCTTGCTTCATT LNA 22nt Table 2: List of cell penetrating peptides (CPPs) as carriers of ASOs. The peptides start with their N-ends.
  • a spectrophotometer measures the density of the bacterial culture (OD 600nm 0.5 - 0.9). If the 240 cell density is greater than 0.9, a dilution (1: 1000) is made to reach the required density of the bacterial cells. After dilution, pre-sterilized cuvettes are used to measure the cell density. The cuvettes contain diluted bacterial cells, MgCl 2 , and sterile water, which are used as a control, while other samples contain and ASO. The optical density is measured on every half hour for eight hours. The ASO is determined as a function of time and optical density of the bacterial 245 cells. The bacteria cells were inoculated in LB medium and incubated at optimal for them
  • Staphylococcus aureus S. aureus
  • Escherichia coli E. coli
  • Bacillus subtilis B. subtilis
  • SEQ ID No: 1 (ASO-1, Figure 2) designed to bind to the complementary sequence of the aptamer domain of the FMN riboswitch is demonstrated in cells of human conditional pathogenic bacteria S. aureus and E. coli and in non-pathogenic to human bacteria B. subtilis.
  • concentration (2000 nM) of SEQ ID No: 1 (ASO-1) (ASO-1)
  • ASO-1 Figure 2 inhibits mostly the bacterial growth of S. aureus as shown by the line with the triangles in Figure 301.
  • the bacterial growth reaches a maximum of around 0.2 optical units after an incubation time of three and a half hours.
  • An ASO-2 is also used, but it does not bind specifically to RNAs in B. subtilis (the line with the rhombs, Figure 301) and E. coli (the line with the upside down triangles, Figure 301) and, therefore should not inhibit the growth of these bacteria. They reach a maximum of 1.3 optical units after an incubation time of four hours.
  • ASO-2 ( Figure 2) serves as a negative control to demonstrate the specific action of SEQ ID No: 1 (ASO-1, Figure 2).
  • the lack of inhibition of bacterial growth of B. subtilis and E. coli in the presence of ASO-2 ( Figure 2) is an indicator that the DNA part from the chimeric oligomer does not hybridize nonspecifically to RNAs expressed in these bacteria.
  • the bacterial growth of the three bacteria, S. aureus (the line with the triangles, Fig 301), E. coli (the line with the circles, Figure 301) and B. subtilis (the line with the rectangles, Figure 301) in the presence of SEQ ID No: 1 (ASO-1, Figure 2) reaches minimum of less than 0.1 optical units after incubation time of seven hours.
  • SEQ ID No: 1 ASO-1, Figure 2
  • This concentration of SEQ ID No: 1 (ASO-1, Figure 2) inhibits the bacterial growth of B. subtilis, E. coli and S. aureus (Figure 302) at the same level as the higher concentration of SEQ ID No: 1 (ASO-1, Figure 2) shown in Figure 302.
  • SEQ ID No: 1 ASO-1, Figure 2
  • the bacterial growth of B. subtilis the line with the rectangles, Figure 302
  • E. coli the line with the circles, Figure 302
  • aureus reaches a maximum of less than 0.3 optical units after an incubation time of three and a half hours (the line with the triangles, Figure 302).
  • the ASO-2 ( Figure 2) does not inhibit the bacterial growth of E. coli (the line with the upside down triangles, Figure 302) and B. subtilis (the line with the rhombs, Figure 302) as they reach a maximum of 1.3 optical units after an incubation time of four hours.
  • SEQ ID No: 1 ASO-1, Figure 2
  • S. aureus the line with the triangles, Figure 302
  • E. coli the line with the circles, Figure 302
  • B. subtilis the line with the rhombs, Figure 302
  • subtilis (the line with the rectangles, Figure 302), reaches minimum of 0.1 optical units after incubation time of seven hours. While in the presence of ASO-2 ( Figure 2) the bacterial growth of E. coli (the line with the upside down triangles, Figure 302) and B. subtilis (the line with the rhombs, Figure 302) reaches minimum of around 0.3 optical units after incubation time of seven hours.
  • subtilis (the line with the rectangles, Figure 303), reaches minimum of 0.1 optical units after incubation time of seven hours. While in the presence of ASO-2 ( Figure 2), the bacterial growth of E. coli (the line with 310 the upside down triangles, Figure 303) and B. subtilis (the line with the rhombs, Figure 303) reaches minimum of around 0.3 optical units after incubation time of seven hours.
  • a minimum inhibitory concentration is the lowest concentration in which an inhibition of the bacterial growth is observed.
  • MIC80 is a standard used to calculate the minimum concentration of the
  • MIC80 is 700 nM, 4.5 ⁇ / ⁇ 1.
  • This concentration includes ASO and pVEC, which is a cell penetrating peptide (CPP). In this concentration it is observed a maximum inhibition of the bacterial growth and a maximum survival of the human
  • the FMN riboswitch is one of the most common riboswitches in bacteria. In Rfam database, the FMN riboswitch was detected in 3098 species of bacteria. Of them, 30 are pathogenic to human bacteria: Bacillus anthracis, Clostridium perfringens, Listeria monocytogenes, Staphylococcus aureus, Clostridium botulinum, Clostridium tetani, Clostridium difficile, Staphylococcus epidermidis, Staphylococcus
  • SEQ ID No: 2 (ASO-1, Figure 5) designed to bind to the complementary sequence of the aptamer domain of the glmS riboswitch is demonstrated in cells of human conditional pathogenic bacteria S. aureus and E. coli and in non-pathogenic to human bacteria B. subtilis.
  • the sequence of SEQ ID No: 2 (ASO-1, Figure 5) is designed to be fully
  • ASO-2 ( Figure 5) is used as a negative control and does not have any influence on the growth of the bacteria.
  • the bacterial growth of S. aureus (the line with the triangles, Figure 602) reaches a maximum of 0.3 optical units after an incubation time of three and a half hours. While the bacterial growth of E. coli (the line with the circles, Figure 602) and B. subtilis (the line with the rectangles, Figure 602) reaches a maximum of 1.3 optical units after an incubation time of five hours. The bacterial growth of the two bacteria, E. coli (the line with the circles,
  • the glmS riboswitch is found in 800 bacterial species, of which 11 are pathogenic to human: Bacillus anthracis, Clostridium perfringens, Listeria monocytogenes, Staphylococcus aureus, Clostridium botulinum, Clostridium tetani, Clostridium difficile, Staphylococcus epidermidis, Staphylococcus
  • Example 3 Inhibitory action of antisense oligonucleotide for SAM-I on the bacterial growth
  • Figure 901 The line with the triangles in Figure 901 shows the bacterial growth of L. monocytogenes in the absence of SEQ ID No: 3 (ASO-1, Figure 8), which reaches a maximum of about 1.3 optical units after an incubation time of three and a half hours.
  • ASO-1 SEQ ID No: 3
  • SEQ ID No: 3 (ASO-1, Figure 8), is observed a maximum of 0.3 optical units after an incubation time of four hours (the line with the circles, Figure 901).
  • S. aureus and L. monocytogenes in the absence of SEQ ID No: 3 (ASO-1, Figure 8), is observed a minimum of bacterial growth of 0.6 optical units after an incubation time of five and a half hours (the lines with the rectangles and with the triangles Figure 901). While, in the presence of SEQ ID No: 3
  • the following samples contain SEQ ID No: 3 (ASO-1, Figure 8) in the concentration of 1000 nM. This concentration of SEQ ID No: 3 (ASO-1, Figure 8) inhibits the bacterial growth ofL. monocytogenes as well as in the higher concentration of SEQ ID No: 3 (ASO-1, Figure 8). While
  • FIG. 902 The line with the upside down triangles shows the bacterial growth of L. monocytogenes in the presence of SEQ ID No: 3 (ASO-1, Figure 8) which reaches a maximum of less than 0.4 optical units after incubation of four hours ( Figure 902).
  • the line with the circles shows the bacterial growth of S. aureus in the presence of SEQ ID No: 3 (ASO-1, Figure 8), which reaches a maximum of less than 0.3 optical units after
  • SEQ ID No: 3 (ASO-1, Figure 8) is 700 nM. At this concentration, an inhibition of the bacterial growth of L. monocytogenes and S. aureus is also observed as in the previous two concentrations of SEQ ID No: 3 (ASO-1) ( Figure 903). In the presence of SEQ ID 445 No: 3 (ASO-1, Figure 8), L. monocytogenes reaches a maximum of bacterial growth of around 0.5 optical units after an incubation time of four hours (the line with the upside down triangles, Figure 903). While S.
  • aureus in the presence of SEQ ID No: 3 (ASO-1, Figure 8) reaches a maximum of bacterial growth by less than 0,4 optical units after an incubation time of three and a half hours (the line with the circles, Figure 903).
  • the lines with the triangles and with the 450 rectangles show the maximum of the bacterial growth of L. monocytogenes as well as of S.
  • MIC 80% is 4.5 ⁇ / ⁇ 1. This concentration includes ASO and pVEC CPP.
  • Riboswitch for SAM-I is found only in Gram-positive pathogenic bacteria. This riboswitch is present in the following 9 pathogenic bacteria: Bacillus anthracis, Clostridium
  • Clostridium tetani Clostridium difficile, Staphylococcus saprophyticus and Streptococcus epidermidis.
  • Example 4 Inhibitory action of antisense oligonucleotide for TPP riboswitch
  • subtilis is in the presence of SEQ ID No: 4 (ASO-1, Figure 11) it reaches a maximum of around 0.4 optical units after an incubation time of three and a half hours (the line with the upside down triangles, Figure 1202). In the absence of SEQ ID No: 4 (ASO-1, Figure 11), the bacterial growth of L. monocytogenes reaches maximum of 1.3 optical units after incubation time of five
  • SEQ ID No: 4 (ASO-1, Figure 11) cannot inhibit the bacterial growth of E. coli because the TPP riboswitch is not present in E. coli and SEQ ID No: 4 (ASO-1, Figure 11) cannot bind specifically and inhibit the bacterial growth.
  • ASO that hybridizes with the
  • MIC80 is 750 nM, 5.2 ⁇ / ⁇ 1. This concentration includes
  • TPP riboswitch is widely spread in human bacterial pathogens. It is
  • Francisella tularensis Helicobacter pylori, Klebsiella pneumoniae, Leptospira interrogans, Legionella pneumophila, Neisseria gonorrheae, Neisseria meningitides, Pseudiminas aeruginosa, Salmonella enterica, Enterobacter sp., Corynebacterium diphtheriae, Enterococcus
  • Figure 1 A general scheme of antisense oligonucleotide based inhibition in bacteria.
  • 590 presents the inhibition of specific mRNAs targeted by three different types and chimeric
  • antisense oligonucleotides All antisense oligonucleotides are coupled with cell penetrating peptides, which penetrate bacterial cells. After mRNA transcription (101), the inhibition of translation of specific protein expression can be achieved by mRNA decay via RNase H (102) or by prevention of mRNA translation (103). These approaches lead to growth inhibition of certain
  • FIG. 1 Specific targeting of FMN aptamer domain in the ribD polycistronic mRNA by a chimeric antisense oligonucleotide (SEQ ID No: 1 (ASO-1)). (201) The chimeric antisense oligonucleotide binds to the complementary sequence of the FMN aptamer domain. (202) After binding of the chimeric antisense oligonucleotide with the FMN aptamer sequence, a double
  • the double- stranded molecule is recognized by the RNase H, which binds it and triggers the enzymatic hydrolysis of mRNA. (204) The enzymatic hydrolysis of mRNA leads to no gene expression of the ribD operon.
  • Figure 3 Inhibition of bacterial growth by antisense oligonucleotide that targets FMN riboswitch mRNA. (301) SEQ ID No: 1 (ASO-1) in the concentration of 2000 nM, binds with
  • ASO-2 (Fig.2) does not inhibit the bacterial growth of B. subtilis (the rhombs) and E. coli (the upside down triangles).
  • SEQ ID No: 1 (ASO-1) in the concentration of 700 nM binds with the mRNA for the FMN riboswitch and inhibits the bacterial growth of B. subtilis (the rectangles), E. coli (the circles) and S. aureus (the triangles).
  • ASO-2 (Fig.2) does not inhibit the bacterial growth of B. subtilis (the rhombs) and E. coli (the upside down triangles).
  • SEQ ID No: 1 (ASO-1) in the concentration of 350 nM has no effect on inhibiting the bacterial growth in B. subtilis (the the rectangles), E. coli (the line with the circles) and S. aureus (the line with the triangles).
  • SEQ ID No: 1 (ASO-1) in the concentration of 150 nM has no effect on inhibiting the bacterial
  • Figure 4 Alignment of the sequences in pathogenic bacteria containing the FMN riboswitch sequence.
  • the pathogenic bacteria S. aureus, S. sapropyticus, S. epidermidis, L. monocytogenes, 625 B. anthracis, S. agalactiae, S. pneumonae, E. coli and the non-pathogenic bacteria B. subtilis contain the FMN riboswitch sequence.
  • SEQ ID No: 2 (ASO-1) in the concentration of 2000 nM inhibits the bacterial growth of S. aureus (the line with the triangles), but it does not inhibit the bacterial growth of B. subtilis (the rectangles) and E. coli (the circles).
  • SEQ ID No: 2 (ASO-1) in the concentration of 1000 nM inhibits the bacterial growth of S. aureus (the triangles), but it does not inhibit the bacterial growth of B. subtilis (the rectangles) and E. coli (the circles).
  • SEQ 640 ID No: 2 (ASO-1) in the concentration of 700 nM inhibits the bacterial growth of S.
  • SEQ ID No: 2 (ASO-1) in the concentration of 350 nM does not inhibit the bacterial growth of S. aureus (the triangles) as same as for the bacterial growth of B. subtilis (the rectangles) and E. coli (the circles).
  • SEQ ID No: 2 (ASO-1) in the concentration of 150 645 nM has no effect on inhibiting the bacterial growth of S. aureus (the triangles) as same as for the bacterial growth of B. subtilis (the rectangles) and E. coli (the circles).
  • Figure 7 Alignment of the sequences in pathogenic bacteria containing the glmS riboswitch sequence. Only S. aureus has a full match with the glmS riboswitch sequence, while the
  • FIG. 8 Specific targeting of SAM-I aptamer domain in the S-box polycistronic mRNA by chimeric antisense oligonucleotide (SEQ ID No: 3 (ASO-1)).
  • SEQ ID No: 3 ASO-1
  • the chimeric antisense oligonucleotide binds to the complementary sequence of the SAM-I aptamer domain.
  • 802 After binding of the chimeric antisense oligonucleotide with the SAM-I aptamer sequence, a double
  • RNA 655 stranded molecule is formed.
  • the double- stranded molecule is recognized by the RNase H which binds it and triggers the enzymatic hydrolysis of RNA.
  • the enzymatic hydrolysis of RNA leads to no gene expression of the S-box operon.
  • Figure 9 Inhibition of bacterial growth by antisense oligonucleotide that targets SAM-I riboswitch mRNA. (901) SEQ ID No: 3 (ASO-1) in concentration of 2000 nM, binds with the
  • SEQ ID No: 3 ASO-1 (the rectangles for S. aureus and the triangles for L. monocytogenes).
  • SEQ ID No: 3 (ASO-1) in concentration of 700 nM binds with the mRNA for the SAM-I riboswitch and inhibits the bacterial growth S. aureus (the circles) and L. monocytogenes (the upside down triangles) in comparison with the bacterial growth without SEQ ID No: 3 (ASO-l)( the
  • SEQ IDONo: 3 (ASO-1) in the concentration of 350 nM has no effect in inhibiting the bacterial growth of S. aureus and L. monocytogenes.
  • SEQ ID No: 3 (ASO-1) in the concentration of 150 nM has no effect in inhibiting the bacterial growth of S. aureus and L. monocytogenes.
  • SEQ ID No: 4 (ASO-1) in concentration of 1000 nM, binds with the mRNA for the TPP riboswitch and inhibits the bacterial growth of L. monocytogenes (the circles) and B. subtilis (the upside down triangles), while the bacterial growth of L.
  • ASO-1 is not inhibited.
  • SEQ ID No: 4 (ASO-1) in the concentration of 350 nM has no effect in inhibiting the bacterial growth of L. monocytogenes (the circles) and B. subtilis (the upside down triangles).
  • SEQ ID No: 4 (ASO-1) in the concentration of 150 nM has no effect in inhibiting the bacterial growth of L. monocytogenes (the circles) and B. subtilis (the upside down triangles).
  • FIG. 13 Alignment of the sequences of pathogenic bacteria containing the TPP riboswitch sequence.
  • the pathogenic bacteria Listeria monocytogenes, Streptococcus pyogenes,
  • Streptococcus pneumonae Streptococcus agalactiae, Mycobacterium tuberculosis,
  • Salmonella enterica contain the TPP riboswitch sequence.
  • Figure 14 Immobilization of thiol-modified ASO to CPP by using of a heterobifunctional cross-linking reagent called 3-(2-pyridyldithio)propionyl hydrazide (PDPH) and 1-Ethyl- 3- (3- dimethylaminopropyl) carbodiimide (EDC). Firstly, the hydrazide group of PDHP reacts with a heterobifunctional cross-linking reagent called 3-(2-pyridyldithio)propionyl hydrazide (PDPH) and 1-Ethyl- 3- (3- dimethylaminopropyl) carbodiimide (EDC). Firstly, the hydrazide group of PDHP reacts with 3-(2-pyridyldithio)propionyl hydrazide (PDPH) and 1-Ethyl- 3- (3- dimethylaminopropyl) carbodiimide (EDC). Firstly, the hydrazide
  • the C-terminus of CPP is activated by 1-Ethyl- 3- (3-dimethylaminopropyl) carbodiimide (EDC)
  • EDC 1-Ethyl- 3- (3-dimethylaminopropyl) carbodiimide
  • a peptide-acyllsourea reactive ester is formed(1502).
  • ASO- hydrazide the peptide-acyllsourea reactive ester forms a peptide bond between the CPP and the ASO-hydrazide is formed (1503).

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Abstract

L'invention concerne la conception, l'ingénierie et les applications de divers oligonucléotides antisens attachés à des peptides de pénétration cellulaire en tant qu'agents antibactériens qui ciblent des ARNm bactériens spécifiques et inhibent la croissance bactérienne (figure 1). La figure 1 présente l'inhibition d'ARNm spécifiques, ciblés par des oligonucléotides antisens chimériques et de trois types différents. Tous les oligonucléotides antisens sont couplés à des peptides de pénétration cellulaire, qui pénètrent dans des cellules bactériennes. Après transcription de l'ARNm (figure 101), l'inhibition de la traduction de l'expression d'une protéine spécifique peut être obtenue par dégradation de l'ARNm par l'intermédiaire de la RNase H (figure 102) ou par empêchement de la traduction de l'ARNm (figure 103). Ces approches conduisent à l'inhibition de la croissance de certaines bactéries pathogènes tel que décrit dans cette demande de brevet (figures 102 et 103).
PCT/IB2017/052402 2017-04-26 2017-04-26 Procédés de création de nouveaux agents antibactériens à l'aide d'oligonucléotides antisens chimériques Ceased WO2018197926A1 (fr)

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BG112506A BG67019B1 (bg) 2017-04-26 2017-05-17 Използване на антисенс олигонуклеотиди с антибактериално действие

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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN109679879A (zh) * 2019-01-25 2019-04-26 石河子大学 一种菌株、菌剂及应用
WO2025006923A3 (fr) * 2023-06-30 2025-05-01 President And Fellows Of Harvard College Analogues d'acide nucléique simple brin destinés à être utilisés en tant que traitement antibactérien résistant aux mutations

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US20130281658A1 (en) * 2010-12-17 2013-10-24 Arrowhead Madison Inc. Peptide-Based In Vivo siRNA Delivery System
US20160083706A1 (en) * 2012-01-18 2016-03-24 Wisconsin Alumni Research Foundation Boronate-mediated delivery of molecules into cells
WO2016146143A1 (fr) * 2015-03-16 2016-09-22 Amal Therapeutics Sa Peptides pénétrant dans les cellules et complexes comprenant ceux-ci
WO2016177900A1 (fr) * 2015-05-06 2016-11-10 Norwegian University Of Science And Technology (Ntnu) Agents antibactériens et utilisation thérapeutique de ceux-ci

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US20130281658A1 (en) * 2010-12-17 2013-10-24 Arrowhead Madison Inc. Peptide-Based In Vivo siRNA Delivery System
US20160083706A1 (en) * 2012-01-18 2016-03-24 Wisconsin Alumni Research Foundation Boronate-mediated delivery of molecules into cells
WO2016146143A1 (fr) * 2015-03-16 2016-09-22 Amal Therapeutics Sa Peptides pénétrant dans les cellules et complexes comprenant ceux-ci
WO2016177900A1 (fr) * 2015-05-06 2016-11-10 Norwegian University Of Science And Technology (Ntnu) Agents antibactériens et utilisation thérapeutique de ceux-ci

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PENCHOVSKY R. ET AL.: "Riboswitch-based antibacterial drug discovery using high- throughput screening methods", EXPERT OPINION ON DRUG DISCOVERY, vol. 8, no. 1, 2013, pages 65 - 82, XP055528929 *
STEIN C. A.: "How to design an antisense oligodeoxynucleotide experiment: a consensus approach.", ANTISENSE AND NUCLEIC ACID DRUG DEVELOPMENT, vol. 8, no. 2, 1998, pages 129 - 132, XP002210583 *

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN109679879A (zh) * 2019-01-25 2019-04-26 石河子大学 一种菌株、菌剂及应用
CN109679879B (zh) * 2019-01-25 2021-07-30 石河子大学 一种菌株、菌剂及应用
WO2025006923A3 (fr) * 2023-06-30 2025-05-01 President And Fellows Of Harvard College Analogues d'acide nucléique simple brin destinés à être utilisés en tant que traitement antibactérien résistant aux mutations

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