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Publication number
WO1999030712A1
WO1999030712A1 PCT/SE1998/002340 SE9802340W WO9930712A1 WO 1999030712 A1 WO1999030712 A1 WO 1999030712A1 SE 9802340 W SE9802340 W SE 9802340W WO 9930712 A1 WO9930712 A1 WO 9930712A1
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WO
WIPO (PCT)
Prior art keywords
poly
drug
2poly
triplex
agent according
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Ceased
Application number
PCT/SE1998/002340
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English (en)
Inventor
Ulrica Sehlstedt
Palok Aich
Jan Bergman
Hans Vallberg
Bengt Nordén
Astrid Gräslund
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Lundblad Leif J I
Original Assignee
Lundblad Leif J I
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Lundblad Leif J I filed Critical Lundblad Leif J I
Priority to JP2000538695A priority Critical patent/JP2002508325A/ja
Priority to AU18990/99A priority patent/AU1899099A/en
Priority to APAP/P/2000/001835A priority patent/AP2000001835A0/en
Priority to IL13652598A priority patent/IL136525A0/xx
Priority to EP98963727A priority patent/EP1037638A1/fr
Publication of WO1999030712A1 publication Critical patent/WO1999030712A1/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

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Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A61K31/4985—Pyrazines or piperazines ortho- or peri-condensed with heterocyclic ring systems
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/12—Antivirals
    • A61P31/14—Antivirals for RNA viruses
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/12—Antivirals
    • A61P31/14—Antivirals for RNA viruses
    • A61P31/18—Antivirals for RNA viruses for HIV
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D487/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00
    • C07D487/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00 in which the condensed system contains two hetero rings
    • C07D487/04—Ortho-condensed systems

Definitions

  • the present invention relates to agents of the general formula I which interact with nucleic acid structures by binding to natural or backbone modified DNA and RNA duplexes and triplexes and hybrides thereof, whereby complexes are formed with said DNA and RNA structures by means of which said structures are stabilized.
  • oligonucleotides are targeted to the unique gene that specifies a disease-related protein and stall transcription by binding to the major groove of the doublestran- ded DNA target.
  • triplexes are, in general, thermodyna- mically less stable than the corresponding duplexes which thus prevents effective use of triplex formation under physiological condition.
  • One approach to circumvent this problem is to design compounds that bind specifically to triplexes and significantly increase the thermal stability of the triplex relative to its corresponding duplex and single strand components. These ligands should then be covalently conjugated to the third strand oligonucleotides to enhance their activity as antige- ne agents, cf. the above article by Thuong & Helene 1993 and the article by Silver et al in J. Amer. Chem. Soc. 1997, 119, pages 263-268.
  • the two most efficient DNA triplex stabilizers reported to date are the ben- zo[g]pyridoindole derivative BgPI (targeted to a mixed sequence oligonucleotide complex, see the article of Escude et al in J.Amer. Chem. Soc. 1995, 1 17, pages 10212- 10219) and 2-naphtyl quinoline derivative targeted to poly(dA).2poly(dT), cf the article of Wilson et al in Biochemistry 1993, 32, pages 10614- 10621.
  • Each of said derivatives provides increments of the triplex-to-duplex transitions by 31°C and 35,6°C respectively.
  • the compound 2,3-dimethyl-6-(dimethyl aminoethyl)-9- hydroxy-6H-indolo[2,3b]quinoxaline (9-OH B220) binds to poly(dA).2poly(dT) under near physiological conditions with an affinity of 1.10 5 M " 1 at 20°C.
  • R ⁇ and R2 which are identical or different, represent lower alkyl with the provisio that the total sum of carbon atoms in R j and R2 is maximum 8 and R j and R2 can also form part of a ring, e.g. piperidino, morpholino or pyrrolidino,
  • R3 and R4, which are identical or different can be hydrogen or lower alkyl with 1 to
  • R5 is hydrogen or methyl, x is 1 or 2.
  • An other object according to the invention is use of the agents as antigene /an tisense enhancing agents when combined with oligonucleotides in form of a covalent or non-covalent complex.
  • a further object of the invention is the use of the agents either alone together with a host ribonucleic acid or combined with an oligonucleotide in form of a covalent or non-covalent complex in preparing drugs for treating HIV and other retroviral infections.
  • the substituted indolo-[2,3b]-quinoxalines are excellent agents for stabilizing natural or backbone modified DNA and RNA triplexes and hybrids thereof by partly intercalative, partly non-intercalative binding.
  • This stabilizing effect can be used in gene expression modification and gene therapy such as use as antigene/ an tisense enhancing agents in combination with the oligonucleotides in form of complex and in gene expression modification as pesticides in plants for plant protection purposes.
  • the agents act by helping to block gene expression of specific genes.
  • RNA polymerase cannot transcribe this gene. Therefore the agents are potentially useful in all types of living cells where transcription of a particular gene should be blocked for a particular reason. A typical reason would be to kill the cell, but other reasons are certainly possible, e.g. to immortalize the cell.
  • the agents are useful e.g. in killing microorganisms while leaving the host less affected. This means that the agents according to the present invention are useful not only in medicine but also in veterinary medicine and pesticide development.
  • the first objective was to establish the conditions at which duplex and triplex forms of the nucleic acid polymers are present as given by the stoichiometry of the single stranded polymers in the sample, without undergoing disproportionation.
  • the drug-triplex interactions under solution environments resembling those of physiological media, i.e. in the presence of both mono- and divalent cations (here
  • Thermal denaturation of the duplexes and triplexes of poly(dA)/poly(dT) and poly(rA)/poly(rU) was studied at different ionic strengths in the absence and presence of drug molecules at or near saturating conditions, defined such that no change in the optical spectrum of the drug is observed upon addition of more nucleic acids.
  • the drug-free duplex structures melt via a single transition.
  • the melting profiles of the triplex structures are generally biphasic; the low temperature step corresponds to the triplex to duplex transition (3- 2), whereas the high temperature transition is assigned to the conversion of the duplex into its component single strand (2 ⁇ T).
  • Figure 1 some selected first derivatives of the obtained melting curves for the DNA and RNA host structures are depicted.
  • the thermal melting temperatures, for each optically detected transition, are summarized in Table 1.
  • poly (dA). poly ⁇ dT) duplex The results presented in Table 1 show, as expected, that the poly (dA). poly (dT) duplex structure is thermodynamically stabilized when the ionic strength of the solution medium increases. Further inspection of the data reveals that an MgCl2 concentration of 2 mM is sufficient to induce disproportionation of a stoichiometric poly(dA).poly(dT) duplex.
  • the triplex structure thus formed is cognate with that of poly(dA).2poly(dT) under comparable salt conditions, as judged from the similar values of the first transition temperature (characterized by poly(dA).2poly(dT) triplex.
  • NaCl results in a destabilization of the triplex to duplex equilibrium as well as the duplex to single strand equilibrium, although the latter to less extent. This is ratio- nalized in terms of a competition between Na and Mg ⁇ for available nucleic acid phosphate binding sites, causing the stabilizing capability of Mg 2+ to be inhibited.
  • poly (r A), poly (rU) duplex The data presented in Table 1 reveals that , as for the DNA hos duplex, the thermodynamic stability of the poly(rA).poly(rU) duplex is enhanced as the ionic strength of the solvent is increased. From the biphasic melting profile obtained at 280 nm it is evident that even in the absence of any added MgCL ⁇ , an
  • NaCl concentration of 100 mM is sufficient to promote heating induced transformation of the poly(rA).poly(rU) duplex into the poly(rA).2poly(rU) triplex and single stranded poly(rA) (spectra not shown).
  • the same behavior is observable in buffers containing 2 mM MgCl2 alone. Melting of poly (r A), poly (rU) at and above these ionic strengths is accompanied by an initial hypochromic absorbance change at 280 nm, corresponding to the so called 2 ⁇ 3 disproportionation transition followed by a hy- perchromic transition attributed to the melting of the poly (r A).2poly(rU). triplex to its single strand components (i.e. the 3->l transition).
  • the stability of the thermally induced poly(rA).2poly(rU) triplex is increased relative to that of the poly(rA).poly(rU) duplex. Therefore, in these cases, the first transition temperature characterizes the 2- 3 transition, while the second one represents the 3—> l transition.
  • the 2- 3 transition is not observable at 260 nm, which is the reason why detection of any disproportionation reaction has to be performed near 280 nm.
  • poly(rA).2poly(rU) triplex Triplex formation by the association of poly(rA) and poly(rU) in a 1:2 stoichiometry is readily accomplished in a buffer containing 50 mM NaCl, as indicated by two sequential steps in the melting profile at 260 nm (Table 1 , spectra not shown). However, the ionic strength has to be increased to at least 100 mM NaCl + 2 mM MgC ⁇ for the formation to be complete. Inspection of
  • Table 1 reveals that the influence of ionic strength on each transition is similar to that just described for the poly(dA).2poly(dT) triplexes, with the same type of competition between mono- and divalent cations for appropriate sites of interaction. Note that at 100 mM Nacl + 5 mM MgC ⁇ , poly(rA).2poly(rU) exhibits a broad single melting transition (depicted in Figure Id, solid curve), which is attributed to a simultaneous dissociation of all three strands involved in the triplex structure. This assignment was confirmed by monitoring the melting process also at 280 and 283.5 nm (data not shown).
  • the melting profile at 283.5 nm where a direct dissociation of poly(rA).2poly(rU) to the single stranded homopolymers cannot be detected, does not contain any transition at all, well in agreement with a disruption of the three stranded complex without any stable intermediate.
  • the melting process was monitored at 280 nm as well (not shown) At this wave- length, the denaturation profile is monophasic at all drug binding ratios with t ⁇ values coinciding with those obtained at 260 nm. In addition, the total hyperchro- micity of the transition remains constant over the whole titration range, which strongly suggests that 9-OH-B220 binds to the poly(rA).2poly(rU) triplex without displacing the major groove-bound third strand from the underlying duplex.
  • the upper and lower panels of Figure 2 show isotropic absorbance (A jso ) and reduced linear dichroism (LD r ) spectra, respectively, of the complexes formed between 9-OH-B220 and poly(dA).poly(dT) (a), poly (rA). poly (rU) (b), poly(dA).2poly(dT) (c) and poly(rA).2poly(rU) (d).
  • a jso isotropic absorbance
  • LD r reduced linear dichroism
  • the absorption spectrum of free 9-OH- B220 consists of three absorption bands in the near UV/vis regions: (i) the highest- energy band is strong with a maximum at 270 nm and has the tendency of a shoulder around 290 nm, (ii) the strong band in the visible region has a peak at 370 nm and a plateau at the high-energy side of the maximum, while (iii) the lowest- energy band, centered around 430 nm, is broad and very weak. In presence of the nucleic acid polymers, these bands are affected differently, depending on the host nucleic acid structure.
  • Spectral effects of this nature are typical for ligands interacting with nucleic acids.
  • Another very useful method for detection and characterization of ligand binding to nucleic acids is linear dichroism. If the absorbing ligands interact with the nucleic acid polymer, an LD signal appears in the drug absorption region when the host nucleic acid becomes oriented, as a result of indirect orientation of the ligand chromophores. If, on the contrary, no binding occurs, no LD from the ligand molecules can be observed.
  • the LD r spectrum of 9-OH-B220 with the poly(dA).poly(dT) duplex (bottom panel of Figure 2a) has a larger negative amplitude in the visible drug absorption bands than in the DNA band around 260 nm. This is indicative of the drug transitions around 380 nm and 430-470 nm being oriented more perpendicular to the helix axis than are on average the base pairs, a phenomenon that has been observed earlier for various intercalating ligands.
  • Table 2 lists the LD r parameters obtained.
  • 9-OH-B220 seems to adopt a nearly perpendicular binding geometry upon interacting with the poly(dA).2poly(dT) triplex, emphasized by the strong negative LD r amplitudes and the calculated effective drug transition moment angles ( Figure 2c, lower panel, and Table 2). Note, however, that binding of 9-OH-B220 to any of the DNA host structures causes a numerical decrease in the LD r amplitude at 260 nm, implying that the orientability of the nucleic acid polymers is reduced (S decreases, Eqn (2) in Experiments, Materials and Methods), which contrasts the usual behavior of classical intercalators. Nevertheless, similar observations have been made with poly(dA)/poly(dT) poljmiers complexed with several intercalating molecules.
  • Binding of 9-OH-B220 to poly(rA).2poly(rU) in the presence of 100 mM NaCl + 5 mM MgCl2 induces LD r spectra with amplitudes that are significantly smaller in the wavelength region above 320 nm than in the UV region, and the calculated average angles between the nucleic acid helix axis and the drug transitions responsible for absorption in the visible region indicate a binding geometry that is nonin- tercalative.
  • Such an interpretation is well in agreement with the observed rather small perturbations of the drug light absorption envelope upon binding to poly(rA).2poly(rU) at this ionic strength ( Figure 2d, top).
  • the LD r signal in the predominantly RNA absorption region around 260 nm decreases upon binding of 9- OH-B220 to any of the RNA host structures, indicating an impaired orientability of the RNA structures as a consequence of drug binding.
  • the purpose of the present experiments was to characterize the nature of interactions between the antiviral quinoxaline derivative 9-OH-B220 and triple helical nucleic acid polymer structures, and to compare with those of their precursor duplexes.
  • Emphasis was put on achieving solution compositions where each of the poly (dA). poly (dT), poly(rA) .poly(rU), poly(dA).2poly(dT) and poly(rA).2poly(rU) complexes were completely formed and remained purely in one state at room temperature.
  • hypochromicity can be utilized to estimate the extent of complex formation.
  • 9-OH-B220 displays a preferential stabilization of the Hoogsteen-paired third strand in poly(dA).2poly(dT) .
  • DNA-interacting agents e.g. the ben- zo[e]- and benzo[g]pyridoindole derivatives BePI (Pilch et al, 1993 J. Mol. Biol. 232, pp 926-946) and BgPI (Escude et al, 1995 loc.cit) all suggested to bind to DNA tri- plexes by intercalation. Groove binding compounds, on the other hand, normally destabilizes DNA triplex structures.
  • the two most efficient DNA triplex stabilizers reported to date are the ben- zo[g]pyridoindole derivative BgPI (targeted to a mixed sequence oligonucleotide. Escude et al, 1995 loc.cit) and a 2-naphtyl quinoline derivative (target to poly(dA).2poly(dT), Wilson et al, 1993b loc.cit), each providing melting temperature increments of the triplex-to-duplex transitions by 31°C and 35.6°C. Both these drugs were, however, investigated in buffer solutions containing significantly smaller amounts of salt than those used in the present work.
  • the ⁇ t ⁇ 3- * 2 being more than 50°C and almost 40°C at a drug to base triplet ratio of 0.50 in buffers containing 100 and 190 mM NaCl, respectively.
  • the drug displays a high discrimination between the triplex and duplex states of poly(dA).2poly(dT) at these ionic strengths, exhibiting fractional ⁇ t m 2 ⁇ 1 / ⁇ t m 3 ⁇ 2 increments of 0.18 and 0.16 at 100 and 190 mM NaCl, respectively.
  • the general conclusion to be drawn from this and other reports is that the size and shape of the ligand chromophore ring system, as well as the positioning of charged side chains, are crucial factors to be considered in developments of DNA triplex stabilizing agents.
  • the compound 9-OH-B220 studied here has a ring system, which shows strong stacking interactions with each of the bases in a triad, and a positively charged side chain that may form favorable interactions with proton acceptor groups in one of the grooves of the triple helical host structure.
  • 9-OH-B220 behaves like a typical intercalator in terms of triplex stabilizing capacity suggests that 9-OH-B220 binds to the poly(dA).2poly(dT) triplex by intercalation, as will be discussed in more detail from a spectroscopic point of view.
  • the above results are indicative of intercalation of the drug chromophore to each duplex structure, but they might also be consistent with a groove binding geometry.
  • important evidence in contradiction of such an interpretation comes from the performed LD measurements.
  • the LD r in the long wavelength region is strongly negative, showing that the orientations of the in-plane polarized transition moments, responsible for absorption in this wavelength range, of 9-OH-B220 when bound to either poly(dA).poly(dT) or poly (rA).
  • poly (rU) are essentially perpendicular to the helix axis of the respective nucleic acid duplex ( Figures 2a and b, lower pa- nels, and Table 2).
  • the LD data argues against a nonspecific electrostatic association of the drug molecules to the phosphate groups at the outside of the triple helical structure, since such surface binding would result in very weak LD signals due to poor average orientation of the interacting ligands, which is evidently not the case here.
  • the major groove of the underlying duplex is blocked due to the presence of the Hoogsteen-paired third strand. Yet, the minor groove is still accessible to interacting agents. Given that the minor groove is wide and shallow in this A-type helical structure, an arrangement of the drug molecules in, and possibly across, this groove constitute an attractive binding geometry model for 9-OH-B220 in complex with poly(rA).2poly(rU). Hence, all findings on the drug-RNA triplex complex point towards homogeneous binding of the 9-OH- B220 molecules in the minor groove of the poly(rA).2poly(rU) host polymer.
  • the deviating drug binding mode in the poly(rA).2poly(rU) triplex suggests that intercalation into the poly (r A), poly (rU) duplex occurs from the major groove side of the helix structure; when the major groove is blocked by a third strand, such as in the poly(rA).2poly(rU) triplex, the drug is hindered from binding intercalatively and is directed to the minor groove of this polymer structure.
  • DNA polymers serve as host nucleic acids, on the other hand, intercalation occurs from the minor groove.
  • a third strand in the major groove of a DNA duplex structure is not expected to severely affect the intercalation process of 9-OH-B220, in agreement with the observations made here.
  • the major groove is less hydrophobic and has a lower electrostatic potential than the minor groove.
  • the positively charged side chain of 9-OH-B220 is likely to preferentially interact with atoms in the major groove of the poly(rA).poly(rU duplex.
  • this groove selectivity of the side chain to be the most probable explanation to the different intercalation directionalities seen for the drug between the RNA- and DNA-polymers.
  • RNA polymers 9-OH-B220 when bound to the RNA polymers 9-OH-B220 exerts a thermally stabilizing effect, being bound intercalatively in the poly(rA).poly(rU) duplex but exhibiting properties characteristic of minor groove binding in the poly(rA).2poly(rU) triplex.
  • the antiviral, low-toxic and highly water soluble compound 9-OH-B220 recognizes and stabilizes both ribonucleotide and deoxyribonucleotide containing helices, which makes it suitable to be conjugated to oligonucleotides to enhance their activity as antigene/ an tisense agents. Furthermore, the present results on particularly the 9- OH-B220-RNA interactions in combination with the known effects of its parent compound B220 on HIV integrase, cf. Swedish patent SE 504,289 makes it a suitable compound for preparing an anti-HIV agent.
  • poly (dT) and poly(rA).poly(rU) were prepared by mixing each strand at a molar ratio of 1: 1. The triplexes were annealed by incubating 1:2 molar ratios of poly (dA)/ poly (dT) and poly(rA)/poly(rU), respectively, at 90°C for 30 min followed by slow cooling to 4°C.
  • LD at a given wavelength is defined as the differential absorption of light polarized with its electric field vector parallel and perpendicular to a macroscopic orientation direction, which equals the flow direction when the sample is oriented by shear gradients.
  • the strength of the LD signal is dependent on the degree of sample orientation, as well as the molar absorptivity and concentration of the absorbing species.
  • the LD is usually normalized by dividing with the absorption spectrum of the corresponding unori- ented sample, so , giving the dimensionless reduced linear dichroism, LD r :
  • DNA and RNA molecules can be assumed to possess an effective cylindrical symmetry about their respecitve helix axis, which makes it possible to express the reduced linear dichroism as a product between an orientation factor S and an optical factor O o r
  • the optical factor depends on the angle ⁇ between the absorbing transition moment of the chromophore and the polynucleotide helix axis; the angle brackets indicate an ensemble average over the angular distribution.
  • LD was measured on a Jasco J-500A spectropolarimeter adapted with an Oxley prism to convert the circularly polarized incident light to linearly polarized.
  • Orientation of the nucleic acid polymers was generated by shear flow in a Couette cell with an outer rotating cylinder. Prior to each measurement, the Couette cell was treated for 30 minutes with a solution of 0.1% DEPC (a strong inhibitor of RNase) in ethanol at 60°C. Thereafter, the Couette cell was rinsed with 25 volumes of ethanol at 75°C to remove traces of DEPC. All experiments were performed at 20°C with a shear gradient of 3100 s .
  • DEPC a strong inhibitor of RNase
  • nucleic acid concentrations were 80 ⁇ M in base pairs/triplets for duplex/triplex polymers, respectively.
  • FIG. 3 Thermal denaturation profiles of free ( ) poly(dA)-2poly(dT) and in the presence of 9-OH-B220 at a drug to base triplet ratio of 0.50 ( ).
  • the triplexes were formed from stoichiometric 1:2 ratios of the single strands in 10 mM sodium cacodylate buffer at pH 7.0 with 1 mM EDTA and (a) 100 mM NaCl (b) 190 mM NaCl.
  • the base triplet concentration was 80 ⁇ M.
  • Nucleic acid polymer [NaCl] (mM) [MgCl 2 ] (mM) m 3 ⁇ 2 ( o C) t m 2 ⁇ i (°C)
  • DNA poly(dA)-poly(dT) 12.5 0 — — 5 555..55 ((8822))
  • the nucleic acid concentrations were 80 ⁇ M in base pairs/triplets and the drug to base pair/triplet ratio was 0.50. All samples were in 10 mM sodium cacodylate (pH 7.0), with 1 mM EDTA, and the measurements were performed either at 260, 280 and/or 283.5 nm.
  • t m 3 ⁇ 2 is assumed to equal t,,, 2- ", and is listed in both columns. Due to heating induced disproportionation of the duplex structures, the low melting temperature characterizes the 2 ⁇ 3 transition while the high melting temperature represents the 3 ⁇ 1 transition (see text for further explanation). Table 2
  • DNA poly(dA)-poly(dT) 12.5 0 -0.054 (-0.115) -0.067 -0.055 0.037 (0.078) 90° 90° poly(dA)-2poly(dT) 100 5 -0.143 (-0.165) -0.119 -0.1 12 0.097 (0.1 12) 76° 74°
  • RNA poly(rA)-poly(rU) 12.5 0 -0.058 (-0.084) -0.066 -0.056 0.046 (0.066) 83° 75°

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Abstract

La présente invention concerne des agents permettant de stabiliser les triplex d'ADN et d'ARN naturels ou à squelette modifié et les hybrides de ceux-ci grâce à des liaisons en partie intercalantes en partie non intercalantes, lesquels agents sont constitués par un composé de la formule (I): dans laquelle n=2 ou 3 et x est égal à 1 ou 2. L'invention se rapporte également à l'utilisation des ligands comme agents activateurs dans les stratégies antigène/antisens et dans la préparation de médicaments destinés au traitement des infections rétrovirales.
PCT/SE1998/002340 1997-12-17 1998-12-16 Agents Ceased WO1999030712A1 (fr)

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Application Number Priority Date Filing Date Title
JP2000538695A JP2002508325A (ja) 1997-12-17 1998-12-16 薬 剤
AU18990/99A AU1899099A (en) 1997-12-17 1998-12-16 Agents
APAP/P/2000/001835A AP2000001835A0 (en) 1997-12-17 1998-12-16 Agents.
IL13652598A IL136525A0 (en) 1997-12-17 1998-12-16 Agents
EP98963727A EP1037638A1 (fr) 1997-12-17 1998-12-16 Agents

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SE9704723A SE9704723D0 (sv) 1997-12-17 1997-12-17 Ligands
SE9704723-7 1997-12-17

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2022002898A1 (fr) * 2020-06-29 2022-01-06 Vironova Medical Ab Dérivés de 6h-indolo(2,3-b)quinoxaline utiles en thérapie, en particulier dans une infection virale

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0238459A1 (fr) * 1986-01-21 1987-09-23 Lundblad, Leif Indoloquinoxalines substituées
WO1996000067A1 (fr) * 1994-06-23 1996-01-04 Leif J. I. Lundblad Inhibiteur
WO1996019996A1 (fr) * 1994-12-27 1996-07-04 Lundblad, Leif, J., I. NOUVELLES APPLICATIONS DES INDOLO-[2,3b]-QUINOXALINES

Patent Citations (3)

* Cited by examiner, † Cited by third party
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