DE19543865A1 - New phosphono:mono:ester oligo:nucleotide analogues - Google Patents

Abstract

Oligonucleotide analogues and related cpds. of formula (I) are new, in which n = 0-100; B = H or OH; 1-20C alkyl, 1-20C alkoxy or 1-20C alkylthio opt. substd. by 6-20C aryl; or aryl or heterocyclyl; where alkyl, aryl and heterocyclyl are opt. substd.; or B = a natural or unnatural nucleobase or a label; A = a single bond, CH2, (CR<2>R<3>)pM(CR<2>R<3>)q or (CR<2>R<3>)rM(CR<2>R<3>)sCY'; or A-B = 1-5 D- or L-amino acids condensed on via the carboxyl group; L = N or R<1>N<+>; R<1> = H or 1-6C alkyl opt. substd. by OH, 1-6C alkoxy, 1-6C alkylthio or NH2; Y' = O, S, CH2, CMe2 or NR<1>; M = a single bond, O, S or NR<1>; R<2> and R<3> = H, OH, 1-6C alkoxy, 1-6C alkylthio, NH2, halogen, or 1-6C alkyl opt. substd. by OH, 1-6C alkoxy or 1-6C alkylthio; p, q, r and s = 0-5; D and G = CR<5>R<6>; R<5> and R<6> = H, 1-6C alkyl, 6-20C aryl, (6-20C)aryl(1-6C)alkyl, OH, 1-6C alkoxy or 1-6C alkylthio, where alkyl and aryl are opt. substd. by SR<1> or NR<1>R<1>; X = O, S or NR<1>; Y = O or S; Z = OR<8>, NR<9>R<1 >0 or XQ; R<8> = H, alkyl, alkenyl, alkynyl, aryl or arylalkyl, where alkyl and aryl are opt. substd.; R<9> and R<10> = H, alkyl, alkenyl, alkynyl, aryl or arylalkyl, where alkyl is opt. substd.; or NR<9>R<10> is a 4- to 7-membered ring; Q and Q' = H, R<8>, opt. modified oligonucleotides or conjugates which (a) favourably affect the properties of antisense oligonucleotides, (b) affect the properties of triplex-forming oligonucleotides, (c) serve as DNA probe labels or (d) bind to or crosslink a target nucleic acid during hybridisation; or Q and Q' alone or together are a single bond in a cyclic molecule; or Q and Q', when neither is hydrogen, can be linked together to form a cyclic molecule.

Description

The present invention relates to novel oligonucleotide analogs with valuable physical, biological and pharmacological properties as well as a Process for their production. Your application relates to the Use as inhibitors of gene expression (antisense oligonucleotides, Ribozymes, Sense Oligonucleotides and Triplex Forming Oligonucleotides), as Probes for the detection of nucleic acids and as tools in the Molecular biology.

Oligonucleotides are increasingly used as inhibitors of Gene expression (J.F. Milligan, M.D. Matteucci and J.C. Martin, J. Med. Chem. 36 (1993) 1923; E. Uhlmann and A. Peyman, Chemical Reviews 90 (1990) 543; S. T. Crooke, Annu. Rev. Pharmacol. Toxicol. 32 (1992) 329).

Antisense oligonucleotides are nucleic acid fragments whose base sequence is complementary to an mRNA to be inhibited. This target mRNA can cellular, viral or other pathogenic origin. As cellular Target sequences come, for example, from receptors, enzymes, Growth factors, immunomodulators, ion channels or oncogenes in Question. Inhibition of Virus Propagation Using Antisense Oligonucleotides have been used, for example, for RSV (Rous Sarcoma Virus), HSV-1 and -2 (Herpes Simplex Virus Type I and II), HIV (Human Immunodeficiency Virus) and influenza viruses. Here one uses oligonucleotides which are used for viral nucleic acid are complementary.

Sense oligonucleotides, however, are designed in their sequence so that they for example nucleic acid binding proteins or nucleic acid processing proteins Bind ("capture") enzymes and thus inhibit their biological activity (C.  H´lène and J. J. Toulm´, Biochim. Biophys. Acta 1049 (1990) 99). As a viral The targets here are, for example, reverse transcriptase, DNA polymerase and To name transactivator proteins. Triplex forming oligonucleotides have generally the DNA as a target and form a after binding to it triple helical structure.

While with the help of the antisense oligonucleotides in general the Processing (splicing etc.) of the mRNA or its translation into the protein are inhibited, triplex forming oligonucleotides inhibit transcription or replication of the DNA (N. T. Thuong, and C. H´lène, Angew. Chem. 105 (1993) 697; Uhlmann and Peyman, Chemical Reviews 90 (1990) 543). It is but also possible single-stranded nucleic acids in a first hybridization with an antisense oligonucleotide to form a double strand bind, which is then in a second hybridization with a triplex forming Oligonucleotide forms a triplex structure. The antisense and triplex Binding regions can either be in two separate oligonucleotides or be housed in an oligonucleotide.

Another application of synthetic oligonucleotides are the so-called Ribozymes that destroy the target RNA due to their ribonuclease activity (D. Castanotto, J.J. Rossi, J.O. Deshler, Critical Rev. Eukar. Gene Expr. 2 (1992) 331).

In DNA diagnostics, nucleic acid fragments are more suitable Labeling as so-called DNA probes or DNA probes for the specific Hybridization to a nucleic acid to be detected. The specific Training of the new double strand is done with the help of the marking, the is preferably not radioactive. This way genetic, malignant, viral or other pathogen diseases to prove.  

For most of the applications mentioned, oligonucleotides are natural in their own right occurring form little or completely unsuitable. You have to chemically do that be modified so that they meet the special requirements. In order to Oligonucleotides in biological systems, for example for the inhibition of Virus multiplication can be used, they must follow Meet prerequisites:

• 1. They must be under in vivo conditions, both in serum and intracellular, have a sufficiently high stability.
• 2. They must be designed so that they cover the cell and nucleus membrane can happen.
• 3. They must be base-specific under physiological conditions Bind to their target nucleic acid to increase the inhibitory effect unfold.

These requirements are not essential for DNA probes; however these oligonucleotides must be derivatized so that detection, for example using fluorescence, chemiluminescence, colorimetry or specific staining, is possible (Beck and Köster, Anal. Chem. 62 (1990) 2258).

A variety of chemical variations of oligonucleotides are known which were synthesized with the aim of better meeting the above requirements fulfill as the unmodified oligonucleotides. The chemical change the oligonucleotides are usually made in such a way that phosphate backbone, Ribose unit or the nucleobases are changed accordingly (Uhlmann and Peyman, Chemical Review 90 (1990) 543). See the modifications there are also those in which both the phosphate bridge and the Sugar unit have been replaced by other groups, for example by "Morpholinonucleoside" oligomers (E.P. Stirchak et al., Nucleic Acids Res. 17 (1989) 6129) or "PNAs" (P.E. Nielsen et al, Bioconj. Chem. 5 (1994) 3). PNA's in particular are characterized by unusually high affinities  Target RNA, but suffer from other unfavorable properties such as lack of solubility or lack of cell penetration (W. Wang et al., Tetrahedron Letters 36 (1995) 1181; M. Egholm et al., In "Innovation and Perspectives in Solid Phase Synthesis, Peptides, Proteins, Nucleic Acids ", Roger Epton, Ed. Mayflower Worldwide Limited, Birminghan, 1994, 145-148).

The task is therefore to create new oligonucleotide analogs with favorable properties to find.

The invention therefore relates to compounds of the formula I.

wherein n is a number from zero to 100;
B independently of one another hydrogen, hydroxy, (C₁-C₂₀) alkyl, (C₁-C₂₀) alkoxy, (C₁-C₂₀) alkylthio, (C₆-C₂₀) aryl, (C₆-C₂₀) aryl- (C₁- C₆) alkyl, (C₆- C₂₀) aryl- (C₁-C₆) alkoxy, (C₆-C₂₀) aryl- (C₁-C₆) alkylthio, means an aromatic group or a heterocyclic group, where alkyl, aryl and / or the aromatic or heterocyclic group optionally one or more times by hydroxy, (C₁-C₄) alkoxy, -NR⁹R¹⁰, -C (O) OH, oxo, -C (O) OR⁸, -C (O) NR⁹R¹⁰, - CN, -F, -Cl, -Br, -NO₂, (C₂-C₆) alkoxyalkyl, -S (O) _m R⁸, - (C₁-C₆) alkyl-S (O) _m R⁸, -NHC (= NH) NHR⁸, -C (= NH) NHR⁸, -NR⁹C (= O) R⁸, = NOR⁸, NR⁹C (= O) OR¹⁰, -OC (= O) NR⁹R¹⁰ and -NR⁹C (= O) NR⁹R¹⁰ may be substituted, or
B represents a natural nucleobase, an unnatural nucleobase or a reporter ligand;
AB can also stand for a D- or L-amino acid condensed on the carboxyl group or for peptides consisting of these amino acids with a length of up to 5 amino acid residues,
L independently of one another N or R¹N⁺, and
R¹ is hydrogen or (C₁-C₆) alkyl, which may be substituted by hydroxy, (C₁-C₆) alkoxy, (C₁-C₆) alkylthio or amino, preferably hydrogen or methyl;
A independently of one another denotes a single bond, a methylene group or a group of the formula IIa or IIb;

Y ′ represents = O, = S, = CH₂, = C (CH₃) ₂ or = N⁺R¹, where R¹ is as defined above;
M represents a single bond, -P-, -S- or -NR¹-, where R¹ is as defined above;
R² and R³ independently of one another are hydrogen, hydroxy, (C₁-C₆) alkoxy, (C₁-C₆) alkylthio, amino, halogen, such as F, Cl, Br or (C₁-C₆) alkyl, which optionally with hydroxy , (C₁-C₆) alkoxy or (C₁-C₆) alkylthio may be substituted, but is preferably hydrogen;
p and q are independently zero to 5;
r and s are independently zero to 5;
D and G represent CR⁵R⁶;
R⁵ and R⁶ independently of one another hydrogen, (C₁-C₆) alkyl, (C₆-C₂₀) aryl, (C₆- C₂₀) aryl- (C₁-C₆) alkyl, hydroxy, (C₁-C₆) alkoxy, ( C₁-C₆) alkylthio, and alkyl and aryl may optionally be substituted with SR¹ or NR¹R¹ ', where R¹ is as defined above and R¹' independently of R¹ has the same meaning as R¹, but R⁵ and R⁶ is preferably hydrogen;
X represents -O-, -S- or -NR¹-, wherein R¹ is as defined above;
Y represents = O or = S;
Z represents -OR⁸, -NR⁹R¹⁰ or X'Q '', where X 'are defined as X and Q''asQ;
R⁸ is hydrogen, (C₁-C₁₈) alkyl, (C₂-C₁₈) alkenyl, (C₃-C₁₈) alkynyl, (C₆-C₁₂) aryl, (C₆-C₁₂) aryl- (C₁-C₆) alkyl means, where alkyl can be substituted one or more times with hydroxy, (C₁-C₄) alkoxy, F, Cl, Br and aryl 1-3 times with hydroxy, (C₁-C₄) alkoxy, (C₁-C₄) alkyl, F, Cl, Br, NO₂, -NR⁹R¹⁰, -C (O) OH, -C (O) O- (C₁-C₆) alkyl, -C (O) NR⁹R¹⁰, may be substituted, but preferably for hydrogen, ( C₁-C₆) alkyl, (C₆-C₁₂) aryl or (C₆- C₁₂) aryl- (C₁-C₆) alkyl, where aryl is simply with (C₁-C₄) alkoxy, (C₁- C₄) - Alkyl, F, Cl, Br, NO₂, may be substituted, particularly preferably hydrogen, (C₁-C₆) alkyl, phenyl or 2- (4-nitrophenyl) ethyl;
R⁹ and R¹⁰ are independently hydrogen, (C₁-C₁₈) alkyl, (C₁-C₁₈) alkenyl, (C₁-C₁₈) alkynyl, (C₆-C₁₂) aryl, (C₆-C₁₂) aryl- (C₁ -C₆) alkyl, where alkyl can be substituted one or more times with hydroxy, (C₁-C₄) alkoxy, F, Cl, Br, or R⁹ and R¹⁰ together with the N atom carrying them can be a 4-7-membered ring form;
Q and Q 'independently of one another are hydrogen or R⁸, stand for conjugates which have a favorable influence on the properties of antisense oligonucleotides or of triple helix-forming oligonucleotides or serve as a label for a DNA probe or in the hybridization of the oligonucleotide analog to the target nucleic acid with binding or attacks crosslinking, or means oligonucleotides which may be unmodified or modified, the following variants being intended to be examples of some modifications (e.g. described in E. Uhlmann and A. Peyman, Chemical Reviews 90 (1990) 543; "Protocols for Oligonucleotides and Analogs ", Synthesis and Proper ties & Synthesis and Analytical Techniques, S. Agrawal, Ed, Humana Press, Totowa, USA 1993):

• a) Complete or partial replacement of the 3'- and / or the 5'-phosphoric diester bridges, for example by phosphorothioate, phosphorodithioate, NR⁴R⁴′-phosphoramidate, boranophosphate, phosphate (C₁-C₂₁) - O-alkyl esters, phosphate [(C₆-C₁₂) aryl (C₁-C₂₁) -O-alkyl] esters, 2,2,2-trichlorodimethylethylphosphonate, (C₁-C₈) alkylphosphonate or (C₆-C₁₂) aryl phosphonate bridges, wherein
  R⁴ and R⁴, independently of one another, are hydrogen, (C₁-C₁₈) alkyl, (C₆-C₂₀) aryl, (C₆-C₁₄) aryl- (C₁-C₈) alkyl or - (CH₂) _c - [NH ( CH₂) _c ] _d -NR⁷R⁷, where c is an integer from 2 to 6 and d is an integer from 0 to 6, and R⁷ is independently hydrogen, (C₁-C₆) alkyl or (C₁-C₄) alkoxy - (C₁-C₆) alkyl, preferably R⁴ and R⁴ 'is hydrogen, (C₁-C₈) alkyl or methoxyethyl, particularly preferably hydrogen, (C₁-C₄) alkyl or methoxyethyl or R⁴ and R⁴' can also together with the nitrogen atom carrying them form a 5-6-membered heterocyclic ring which can additionally contain a further heteroatom from the series O, S, N;
• b) complete or partial replacement of the 3'- or 5′-phosphoric acid diester bridges through "dephospho" bridges (see e.g. Uhlmann and Peyman in "Methods in Molecular Biology", Vol. 20, "Protocols for Oligonucleotides and Analogs ", S. Agrawal, Ed., Humana Press, Totowa 1993, Chapter 16, 355ff), for example by formacetal, 3′-thioformacetal, Methyl hydroxylamine, oxime, methylene dimethyl hydrazo, dimethylene sulfone or Silyl groups;
• c) complete or partial replacement of the sugar phosphate backbone, for example by "morpholinonucleoside" oligomers (E.P. Stirchak et al., Nucleic Acids Res. 17 (1989) 6129) or "PNAs" (P.E. Nielsen et al, Bioconj. Chem. 5 (1994) 3), or PNA-DNA hybrids such as in DE-P 44 08 528.1 (HOE 94 / F 057);
• d) complete or partial replacement of the β-D-2′-deoxyribose units, for example by α-D-2'-deoxyribose, L-2'-deoxyribose, 2′-F-2′-deoxyribose, 2′-O- (C₁-C₆) alkyl-ribose, 2′-O- (C₂-C₆) alkenyl-ribose, 2′-NH₂-2′-deoxyribose, β-D-xylofuranose, α-arabinofuranose, 2,4-dideoxy-β-D-erythro hexopyranose, and carbocyclic (e.g. Froehler, J. Am. Chem. Soc. 114 (1992) 8320) and open-chain sugar analogs (e.g. Vandendriessche et al., Tetrahedron 49 (1993) 7223) or bicyclo sugar analogs (e.g. M. Tarkov et al., Helv. Chim. Acta 76 (1993) 481);
• e) complete or partial replacement of the natural nucleoside bases, for example by 5- (hydroxymethyl) uracil, 5-aminouracil, pseudouracil, Dihydrouracil, 5- (C₁-C₆) alkyl uracil, 5- (C₂-C₆) alkenyl uracil, 5- (C₂-C₆) alkynyl  uracil (described, for example, in Gutierrez et al., J. Am. Chem. Soc. 11 6 (1994) 540 or Sagi et al., Tetrahedron Lett. 34 (1993) 2191), 5- (C₁-C₆) alkyl cytosine, 5- (C₂-C₆) alkenyl cytosine, 5- (C₂-C₆) alkynyl cytosine (Gutierrez et al., J. At the. Chem. Soc. 116 (1994) 540 or Sagi et al., Tetrahedron Lett. 34 (1993) 2191), 5-fluorouracil, 5-fluorocytosine, 5-chlorouracil, 5-chlorocytosine, 5-bromouracil, 5-bromocytosine or 7-deaza-7 substituted purines (described for example in Seela, Nucl. Acids Res. 20 (1992) 2297; Heterocycles 34 (1992) 229).

Q and Q ′ can also stand for conjugates which have the properties of Antisense oligonucleotides or triple helix-forming oligonucleotides (such as for example cell penetration, nuclease degradation, affinity for target RNA / DNA, Pharmacokinetics) or as a label for a DNA probe serve or in the hybridization of the oligonucleotide analog to the target Nucleic acid attacks them with binding or cross-linking. Examples of this are conjugates with poly-lysine, with intercalators such as pyrene, acridine, phenazine, Phenanthridine, with fluorescent compounds such as fluorescein, with cross Linkers such as psoralen, azidoproflavin, with lipophilic molecules such as (C₁₂-C₂₀) alkyl, with lipids like 1,2-di-hexadecyl-rac-glycerin, with steroids like Cholesterol or testosterone, with vitamins like vitamin E, with poly or oligo ethylene glycol, with (C₁₂-C₁₈) alkyl phosphate diesters, with -O-CH₂-CH (OH) -O- (C₁₂-C₁₈) alkyl. Preferred are conjugates with lipophilic molecules such as (C₁₂-C₂₀) alkyl, with steroids like cholesterol or testosterone, with poly or Oligoethylene glycol, with vitamin E, with intercalators such as pyrene, with (C₁₄-C₁₈) alkyl- Phosphate diesters, with -O-CH₂-CH (OH) -O- (C₁₂-C₁₆) alkyl.

The preparation of such oligonucleotide conjugates is known to the person skilled in the art (see e.g. B. Uhlmann & Peyman, Chem. Rev. 90 (1990) 543; M. Manoharan in "Antisense Research and Applications," Crooke and Lebleu, Eds., CRC Press, Boca Raton, 1993, Chapter 17, pp. 303ff. and EP-A 0 552 766). Farther can the oligonucleotides at 3 'or at the 5'-end 3'-3'- and 5'-5'-inversions (described, for example, in M. Koga et al., J. Org. Chem. 56 (1991) 3757) carry.  

Aromatic groups are, for example, phenyl, naphthyl, pyrenyl, Anthracenyl, phenanthryl, biphenyl, binaphtyl, tetracenyl, pentacenyl, Hexacenyl, triphenylenyl, chrysenyl or benzopyrenyl.

Examples of heterocyclic groups include chromanyl, chromenylium-1-yl, Furanyl, isochromanyl, isochromenyl, isoquinolyl, piperazinyl, quinolinyl, Pyridinyl, pyrrolidinyl, imidazyl, tetrahydrofuranyl, aziranyl, oxiranyl, Thiophenyl, pyrimidinyl, thielanyl, thiazolyl, azepinyl, pyrrolyl, Tetrahydropyrrolyl, benzofuranyl, indolyl, isoindolyl, isatinyl, dioxindolyl, Indoxylyl, coumarinyl, coumaronyl, carbazolyl, pyrazolyl, pyrrolyl, indazolyl, Oxazolyl, isoxazolyl, thiazolyl, 1,2,4-triazolyl, 1,2,3-triazolyl, tetrazolyl, Pentazolyl, piperidinyl, pyradizinyl, phenazinyl, phenoxazinyl, phenothiazinyl, Morpholinyl, thiazinyl, benzodiazepinyl, purinyl, xanthinyl, hypoxanthine, Theophyllinyl, Theobrominyl, Caffeinyl, Pteridinyl, Pterinyl, Pteridinyl, Alloxazinyl and understand Nortropinyl.

Under natural nucleobases z. B. uracil, cytosine, 5-methyluracil, Adenine and guanine and under unnatural nucleobases are e.g. B. 5-nitroindole, 5- (hydroxymethyl) uracil, 5-aminouracil, pseudouracil, dihydrouracil, 5- (C₁-C₆) alkyl uracil, 5- (C₂-C₆) alkenyl uracil, 5- (C₃-C₆) alkynyl uracil, 5- (C₁-C₆) alkyl cytosine, 5- (C₂-C₆) alkenyl cytosine, 5- (C₃-C₆) alkynyl cytosine, 5-fluorouracil, 5-fluorocytosine, 5-chlorouracil, 5-chlorocytosine, 5-bromouracil, 5-bromocytosine and 7-deaza-7-substituted purines, such as 7-deaza-7- (C₃-C₇) -alkynylguanine, 7-deaza- 7- (C₃-C₇) alkynyladenine, 7-deaza-7- (C₂-C₇) alkenylguanine, 7-deaza-7- (C₂-C₇) -al kenyladenine, 7-deaza-7- (C₁-C₇) alkyl guanine, 7-deaza-7- (C₁-C₇) alkyl adenine, 7-Deaza-7-bromoguanine and 7-Deaza-7-bromadenin understood.

Preferably, unnatural nucleobases are 5- (C₁-C₆) alkyl uracil, 5- (C₂-C₆) alkenyl uracil, 5- (C₃-C₆) alkynyl uracil, 5- (C₁-C₆) alkyl cytosine, 5- (C₂-C₆) alkenyl cytosine, 5- (C₃-C₆) alkynyl cytosine, 5-fluorouracil, 5-fluorocytosine, 5-chlorouracil, 5-chlorocytosine, 5-bromouracil, 5-bromocytosine, or 7-deaza-7 substituted purines  such as 7-deaza-7- (C₃-C₇) alkynylguanine, 7-deaza-7- (C₃-C₇) alkynyladenine, 7-deaza- 7- (C₂-C₇) alkenylguanine, 7-deaza-7- (C₂-C₇) alkenyladenine, 7-deaza-7- (C₁-C₇) -al kylguanine, 7-deaza-7- (C₁-C₇) alkyladenine, 7-deaza-7-bromoguanine and 7-deaza- 7-bromadenine, particularly preferably for 5- (C₃-C₆) alkyl uracil, 5- (C₂-C₆) alkenyl uracil, 5- (C₃-C₆) alkynyl uracil, 5- (C₁-C₆) alkyl cytosine, 5- (C₂-C₆) alkenyl cytosine, 5- (C₃-C₆) alkynyl cytosine or 7-deaza-7-substituted purines, and whole particularly preferred for 5-pentynylcytosine, 5-hexynyluracil, 5-hexynylcytosine, 7-deaza-7-propynylguanine, 7-deaza-7-propynyladenine, 7-deaza-7-methylguanine, 7-deaza-7-methyladenine, 7-deaza-7-propynyladenine, 7-deaza-7-bromoguanine, 7-deaza-7-bromadenine.

Reporter ligands are, for example, fluorescein, biotin, acridine, phenanthroline, Phenanthridine and eosin.

Unless otherwise stated, the following are particularly mentioned among D- or L-amino acids (cf. Schröder, Lübke, Peptides, Volume 1, New York 1965, Pages XXII-XXIII; Houben-Weyl, Methods of Organic Chemistry, Volume XV / 1 and 2, Stuttgart 1974):
Aad, Abu, γAbu, ABz, 2ABz, εAca, Ach, Acp, Adpd, Ahb, Aib, βAib, Ala, βAla, ΔAla, Alg, All, Ama, Amt, Ape, Apm, Apr, Arg, Asn, Asp, Asu, Aze, Azi, Bai, Bph, Can, Cit, Cys, Cyta, Daad, Dab, Dadd, Dap, Dapm, Dasu, Djen, Dpa, Dtc, Fel, Gln, Glu, Gly, Guv, hAla, hArg, hCys, hGln, hGlu, His, hlle, hLeu, hLys, hMet, hPhe, hPro, hSer, hThr, hTrp, hTyr, Hyl, Hyp, 3Hyp, Ile, Ise, Iva, Kyn, Lant, Lcn, Leu, Lsg, Lys, βLys, ΔLys, Met, Mim, Min, hArg, Nle, Nva, Oly, Orn, Pan, Pec, Pen, Phe, Phg, Pic, Pro, ΔPrn, Pse, Pya, Pyr, Pza, Qin, Ros, Sar, Sec, Sem, Ser, Thi, βThi, Thr, Thy, Thx, Tia, Tle, Tly, Trp, Trta, Tyr, Val etc., whose abbreviation stands for the rest in the L-form without a stereo descriptor,
or also cyclic amino acids, such as. B. pyrrolidine-2-carboxylic acid; Piperidine-2-carboxylic acid; 1,2,3,4-tetra hydroisoquinoline-3-carboxylic acid; Decahydroisoquinoline-3-carboxylic acid;
Octahydroindole-2-carboxylic acid; Decahydroquinoline-2-carboxylic acid;
Octahydrocyclopenta [b] pyrrole-2-carboxylic acid;
2-azabicyclo [2.2.2] octane-3-carboxylic acid;
2-azabicyclo [2.2.1] heptane-3-carboxylic acid;
2-azabicyclo [3.1.0] hexane-3-carboxylic acid;
2-azaspiro [4.4] nonane-3-carboxylic acid;
2-Az aspiro [4.5] decane-3-carboxylic acid;
Spiro [(bicyclo [2.2.1] heptane) -2,3-pyrrolidine-5-carboxylic acid];
Spiro [(bicyclo [2.2.2] octane) -2,3-pyrrolidine-5-carboxylic acid];
2-azatricyclo [4.3.0.1 ^6,9 ] decane-3-carboxylic acid;
Decahydrocyclohepta [b] pyrrole-2-carboxylic acid;
Decahydrocycloocta [b] pyrrole-2-carboxylic acid;
Octahydrocyclopenta [c] pyrrole-2-carboxylic acid;
Octahydroisoindole-1-carboxylic acid;
2,3,3a, 4,6a-hexahydrocyclopenta [b] pyrrole-2-carboxylic acid;
2,3,3a, 4,5,7a-hexahydroindole-2-carboxylic acid;
Tetrahydrothi azole-4-carboxylic acid;
Isoxazolidine-3-carboxylic acid; Pyrazolidine-3-carboxylic acid;
Hydroxypyrrolidine-2-carboxylic acid; which can all be optionally substituted:

US-A 4,344,949, US-A 4,374,847, US-A 4,350,704, EP-A 29 488, EP-A 31 741, EP-A 46 953, EP-A 49 605, EP-A 49 658, EP-A 50 800, EP-A 51 020, EP-A 52 870, EP-A 79 022, EP-A 84 164, EP-A 89 637, EP-A 90 341, EP-A 90 362, EP-A 105 102, EP-A 109 020, EP-A 111 873, EP-A 271 865 and EP-A 344 682.

Alkyl and radicals derived therefrom, such as, for example, alkoxy and alkylthio can be branched, unbranched or cyclic, saturated or one or more be unsaturated.

Preferred compounds of the formula I are those in which
n represents a number from zero to 50;
B independently represents a natural nucleobase or an unnatural nucleobase;
LN means;
A represents a group of formula IIb, wherein
r = 1 and s zero, and R², R³ = H and Y ′ = O and M represent a single bond;
D and G mean CHR⁵;
R⁵ represents hydrogen;
X represents -O-;
Y = O;
Z represents hydroxy, methoxy, ethoxy, (4-nitrophenol) ethoxy, propoxy, isoPropoxy, butoxy, pentoxy, phenoxy or allyloxy;
Q and Q 'independently of one another represent hydrogen, Rigon or oligonucleotides, which can be unmodified and modified, where

• a) The 3'- and / or 5'-phosphoric acid diester bridges completely or partially by phosphorothioate, phosphorus dithioate, NR⁴R ^{4 '} phosphoramidate, N3' → P5'-phosphorus amidate (for example described in Gryaznov et al., J. Am Chem. Soc. 116 (1994) 3143), phosphate-O-methyl ester, phosphate-O-ethyl ester, phosphate-O-isopropyl ester, methylphosphonate or phenylphosphonate bridges are replaced;
• b) one, two or three 3'- or 5-phosphoric diester bridges to the Pyrimidine positions and at the 5'-end and / or at the 3'-end by formacetals and / or 3'-thioformacetals are replaced;
• c) the sugar phosphate backbone completely or partially by "PNAs" or PNA-DNA hybrid is replaced;
• d) the β-D-2'-deoxyribose units completely or partially by 2'-F-2'-deoxyribose, 2'-O- (C₁-C₆) alkyl ribose, 2'-O- (C₂-C₆) alkenyl ribose, 2'-NH₂-2'-deoxy ribose are replaced;
• e) the natural nucleoside bases completely or partially by 5- (C₁-C₆) alkyl uracil, 5- (C₂-C₆) alkenyl uracil, 5- (C₂-C₆) alkynyl uracil, 5- (C₁-C₆) alkyl cytosine, 5- (C₂-C₆) alkenyl cytosine, 5- (C₂-C₆) alkynyl cytosine, 5-fluorouracil, 5-fluorocytosine, 5-chlorouracil, 5-chlorocytosine, 5-bromouracil, 5-bromocytosine, 7-Deaza-7- (C₂-C₇) alkynyl guanine, 7-Deaza-7- (C₂-C₇) alkynyladenine, 7-Deaza-7- (C₂-C₇) -al kenylguanine, 7-deaza-7- (C₂-C₇) alkenyladenine, 7-deaza-7- (C₁-C₇) -al kylguanine, 7-deaza-7- (C₁-C₇) alkyladenine, 7-deaza-7-bromoguanine, 7-deaza-  7-bromadenin are replaced.

Compounds of the formula I in which
n represents a number from 0 to 30;
Q and Q ′ independently of one another are hydrogen, R⁸, where R⁸ is H, (C₁-C₆) alkyl, phenyl or 2- (4-nitrophenylethyl), or are oligonucleotides which can be unmodified and modified, where

• a) The 3'- and / or 5'-phosphoric diester bridges completely or partially by phosphorothioate, phosphorodithioate or methylphosphonate bridges are replaced;
• b) one, two or three 3'- or 5-phosphoric diester bridges on 5'- and on 3'-end are replaced;
• c) the sugar phosphate backbone completely or partially by "PNAs" or PNA-DNA hybrid is replaced;
• d) the β-D-2'-deoxyribose units completely or partially by 2'-F-2'-deoxy ribose, 2'-O- (C₁-C₄) alkyl-ribose, 2'-O- (C₂-C₄) alkenyl-ribose, 2'-NH₂-2'-deoxy ribose are replaced;
• e) the natural nucleoside bases completely or partially by 5- (C₃-C₆) alkyl uracil, 5- (C₂-C₆) alkenyl uracil, 5- (C₂-C₆) alkynyl uracil, 5- (C₁-C₆) alkyl cytosine, 5- (C₂-C₆) alkenyl cytosine, 5- (C₂-C₆) alkynyl cytosine, 7-deaza-7- (C₂-C₇) -al kinylguanine, 7-deaza-7- (C₂-C₇) alkynyladenine, 7-deaza-7- (C₂-C₇) alkenylguanine, 7-deaza-7- (C₂-C₇) alkenyladenine, 7-deaza-7- (C₁-C₇) alkylguanine, 7-deaza- 7- (C₁-C₇) alkyladenine, 7-deaza-7-bromoguanine, 7-deaza-7-bromadenine are replaced.

Compounds of the formula I in which
n represents a number from 0 to 25;
B independently represents a natural nucleobase;
Z represents hydroxy, ethoxy, (4-nitrophenol) ethoxy or phenoxy;
Q and Q ′ independently of one another are hydrogen, R⁸, where R⁸ is H, (C₁-C₆) alkyl, phenyl or 2- (4-nitrophenylethyl), or are oligonucleotides which can be unmodified and modified, where

• a) The 3'- and / or 5'-phosphoric diester bridges completely or partially are replaced by phosphorothioate bridges;
• c) the sugar phosphate backbone completely or partially by "PNAs" or PNA-DNA hybrid is replaced;
• d) the β-D-2'-deoxyribose units completely or partially by 2'-O-methyl, 2'-O-allyl, 2'-O-butylribose are replaced;
• e) completely or partially through the natural nucleoside bases 5-hexynylcytosine, 5-hexynyluracil, 5-hexynylcytosine, -deaza-7-propynylguanine, 7-deaza-7-propynyladenine, 7-deaza-7-methylguanine, 7-deaza-7-methyladenine, 7-Deaza-7-propinyladenine, 7-Deaza-7-bromoguanine, 7-Deaza-7-bromadenine replaced are.

The invention further relates to compounds of the formula I in which Q and Q 'are linked, d. H. form a cyclic molecule, the Possibility should apply that Q and Q 'together result in a single bond.  

The synthesis of such compounds can be carried out analogously to the methods described such as Gao et al., Nucl. Acids Res. 23 (1995) 2025 or Wang and Kool, Nucl. Acids Res. 22 (1994) 2326.

Another object of the invention are oligonucleotides or modified Oligonucleotides, for example PNA's, in which compounds of the formula I am 3'-end or at the 5'-end or at the 5'- and at the 3'-end are installed.

The oligonucleotides are linked to the compounds of the formula I. preferably via the 5'- or 3'-hydroxy group of the nucleotide building blocks, likewise via a phosphonic acid monoester bond. In formulas XVIII and XIX the linkage with oligonucleotides is exemplified.

R¹⁷ here stands for H, OH, F, 2'-O- (C₁-C₆) alkyl, 2'-O- (C₂-C₆) alkenyl, preferably for H or methoxy or O-allyl, particularly preferably H. All other Variables are explained above.

Formula XX and Formula XXI, in which the variables have the above meaning, exemplify the link with PNA's.

The combinations of the compounds according to the invention (abbreviated PMENA) with oligonucleotides or modified oligonucleotides, such as PNA's or other modifications as described above, are to be clarified again schematically (OLIGO stands for unmodified or modified oligonucleotides):
Examples of such combinations are:
5′-OLIGO - PMENA
5′-PMENA - OLIGO
5′-OLIGO - PMENA - OLIGO
5′-OLIGO - (PMENA - OLIGO) _a (a = 1-20)
5′-PMENA - OLIGO - PMENA
5′-PMENA - (OLIGO - PMENA) _a (a = 1-20).

The synthesis of these combined compounds takes place in such a way that, depending on the molecule first with the synthesis of the PMENA building blocks, which are described below is started, which is then coupled to the oligonucleotide building blocks  will. The oligonucleotides are known to those skilled in the art Methods (Sonveaux, Bioorganic Chemistry 14 (1986) 274ff) Solid phase synthesis or by solution synthesis as monomer units or coupled by block condensation. The condensations are optional according to the amidite method, the H-phosphonate method or the Phosphortriester method (Sonveaux, Bioorganic Chemistry 14 (1986) 274ff). Conversely, if PMENA modules are linked to OLIGO modules, this is done this is preferred according to the method described in f₁). The conjugation with PNA-Bau stones are done in the same way or if (monomeric or oligomeric) PNA modules are coupled to PMENA modules under the Methods known to those skilled in the art of peptide synthesis or ester synthesis.

The invention further relates to a process for the preparation of compounds of formula I, which is characterized in that
a₁) compounds of formula III

wherein
D, G, L and X have the meanings given above and
S¹ stands for a suitable protective group, such as, for example, dimethoxytrityl, monomethoxytrityl, trityl, pixyl, tert-butoxycarbonyl or fluorenylmethyloxycarbonyl, preferably monomethoxytrityl or tert-butoxycarbonyl,
with compounds of formula IV

wherein
R⁵ and R⁶ have the meanings given above,
in a suitable organic solvent, for example in methanol, ethanol, isopropanol, butanol, acetonitrile, dichloromethane (DCM), chloroform, benzene, dimethylformamide (DMF), dimethyl sulfoxide (DMSO), diethyl ether, ethyl acetate (EE), tetrahydrofuran (THF) , N-methylpyrrolidone, petroleum ether, xylene or toluene or mixtures of suitable solvents, preferably in methanol or ethanol, at temperatures from 0 ° C. to 100 ° C., preferably at 10 to 50 ° C., to give compounds of the formula Va or Vb

the choice of reaction conditions known to those skilled in the art (e.g. in SR Sandler, W. Karo "Organic Functional Group Preparations", Vol. II, Second Edition, Academic Press, London, 1986, Chapter 12 ("Imines ")) care must be taken to ensure that they are compatible with the protective group S1, ie if, for example, an acid-labile protective group such as the monomethoxytrityl protective group is selected, acid addition should be avoided in the reaction,
b₁) Compounds of the formula Va or Vb with compounds of the formula VIa or VIb, preferably with compounds of the formula VIa

wherein
Y is as defined above,
X ′ and X ′ ′ are independently defined as X,
S² and S³ independently of one another represent protective groups, such as, for example, methyl, ethyl, phenyl, 2-chlorophenyl, 2,5-dichlorophenyl, 2,4-dichlorophenyl, 4-nitrophenyl, 4-methoxyphenyl, 2-cyanoethyl, 2- (4- Nitro phenyl) ethyl, allyl, benzyl, 2,2,2-trichloro-1,1-dimethylethyl, 4-methoxybenzyl, 2,2,2-trichloroethyl, 8-hydroxyquinoline or other phosphate protecting groups as are known to the person skilled in the art (Sonveaux , Bioorganic Chemistry 14 (1986) 274ff), but preferably methyl, ethyl, phenyl, 2- (4-nitrophenyl) ethyl, allyl, 2,2,2-trichloroethyl, and
L¹ represents a leaving group, preferably (C₁-C₄) alkyl,
in a suitable organic solvent, for example in methanol, ethanol, isopropanol, butanol, acetonitrile, benzene, DMF, DMSO, DCM, EE, chloroform, diethyl ether, THF, N-methylpyrrolidone, petroleum ether, xylene or toluene or mixtures of suitable solvents, preferably in THF, at temperatures from 0 ° C to 100 ° C, preferably at 50 to 80 ° C, optionally with the addition of bases, such as tri- (C₁-C₆) alkylamine, N-alkylmorpholine, pyridine, N, N -Dimethylaminopyridine, butyllithium, lithium diisopropylamide (LDA), sodium hydride, sodium amide, potassium carbonate, cesium carbonate, potassium tert-butoxide or complex bases such as sodium amide- R¹¹ONa, where R¹¹ is (C₂-C₆) -alkyl or CH₃CH₂-O-CH₂CH₃, or uncharged, peralkylated polyamino-phosphazene bases (Schwesinger, Nachr. Chem. Techn. Lab. 38 (1990) 1214; Angew. Chem. 99 (1987) 1212), but preferably without base addition,
converts to compounds of formula VII

wherein
D, G, L, R⁵, R⁶, S¹, S², S³, X, X ′, X ′ ′ and Y are as defined above;
c₁) compounds of the formula VII with compounds of the formula VIII,

their synthesis z. B. Dueholm et al., J. Org. Chem. 59 (1994) 5767 and
wherein
A has the meaning given above,
B ^{PR has} the same meaning as B, but may be in a protected form, ie if B stands for a natural or unnatural nucleobase, B ^PR stands for the nucleobases whose amino or hydroxyl groups are protected by suitable known protective groups, such as, for example the para-nitrophenylethyl group, the benzoyl group, the allyl group and the para- (t-butyl) benzoyl group for the hydroxyl group and the acetyl, benzoyl, para- (t-butyl) benzoyl, para- ( Methoxy) benzoyl group, para-nitrophenylethyloxycarbonyl group, isobutyryl group, para- (t-butyl) phenylacetyl group, N, N-dimethylformamidino, fluorenylmethyloxycarbonyl group, benzyloxycarbonyl group, phenoxyacetyl group for the amino group or others protective groups customary for nucleobases in oligonucleotide chemistry (Sonveaux, Bioorganic Chemistry 14 (1986) 274ff; Beaucage, Tetrahedron 49 (1993) 2223ff), preferably for B ^PR :

wherein
R¹² is hydrogen, 1-propynyl, 1-butynyl, 1-pentynyl or 1-hexynyl, especially hydrogen, 1-propynyl or 1-hexynyl; and
R¹³ is hydrogen, diphenylcarbamoyl or 2- (4-nitrophenyl) ethyl and
R¹⁴ means acetyl, benzoyl, para- (t-butyl) benzoyl, para- (methoxy) benzoyl, para-nitrophenylethyloxycarbonyl, isobutyryl, para- (t-butyl) phenylacetyl, benzyloxycarbonyl or phenoxyacetyl, and
L² represents a leaving group known to the person skilled in the art, such as Cl, Br, O-SO₂methyl, O-SO₂ trifluoromethyl, OTosylate, O-CatF₆ or, if A has the meaning of formula IIb, can also represent OH;
in a suitable organic solvent, for example in acetonitrile, benzene, DMF, DMSO, DCM, EE, chloroform, diethyl ether, tetramethyl urea, THF, N-methylpyrrolidone, petroleum ether, xylene, or toluene or mixtures of suitable solvents, preferably in DMF, at temperatures of -20 ° C to 100 ° C, preferably at 0 to 50 ° C, optionally with the addition of bases such as tri- (C₁-C₆) alkylamine, N-alkylmorpholine, pyridine, N, N-dimethylaminopyridine, butyllithium, lithium diisopropylamide (LDA), sodium hydride, sodium amide, potassium carbonate, cesium carbonate, potassium tert-butoxide or complex bases such as sodium amide R¹¹ONa, where R¹¹ is (C₂-C₆) alkyl or CH₃CH₂-O-CH₂CH₃ or uncharged, peralkylated polyamino-phosphazene -Basen (Schwesinger, Nachr. Chem. Techn. Lab. 38 (1990) 1214; Angew. Chem. 99 (1987) 1212), if A is formula IIb and L² is OH, preferably with the addition of triethylamine, diisopropylethylamine or N -Ethylmorpholine or he without base addition and with the addition of a coupling reagent customary for the formation of peptide bonds,
converts to compounds of formula IX

wherein
A, B ^PR , D, G, L, R⁵, R⁶, S¹, S², S³, X, X ′, X ′ ′ and Y are as defined above;
d₁) from compounds of formula IX the protective group S³ by known methods (z. B. Greene, Wuts, Protective Groups in Organic Synthesis, J. Wiley & Sons, New York 1991), so z. B. for compounds of formula IX, in which S² and S³ are 2- (4-nitrophenyl) ethyl by treatment with 0.1M 1,8-diazabicyclo [5,4,0] undec-7-ene (DBU) in pyridine or acetonitrile at room temperature or for compounds of the formula IX in which S² and S³ are phenyl or ethyl by treatment with aqueous ammonia or for compounds of the formula IX in which S² is 2- (4-nitrophenyl) ethyl and S³ is allyl by treatment with Pd [P (C₆H₅) ₃] ₄ and triphenylphosphine in DCM (Hayakawa et al., J. Org. Chem. 58 (1993) 5551), or for compounds of the formula IX in which S² is 2- (4th -Nitrophenyl) ethyl and S³ stands for allyl by treatment with 0.5M DBU in pyridine or acetonitrile or for compounds of the formula IX in which S² stands for 2-cyanoethyl and S³ for allyl stands for treatment with triethylamine in pyridine, or for compounds of the formula IX in which S² is 2- (4-nitrophenyl) ethyl and S³ is 2,2,2-trichloro-1,1-dimethylethyl by Beh treatment with tributylphosphine,
whereby compounds of formula X are obtained

wherein
A, B ^PR , D, G, L, R⁵, R⁶, S¹, S², X ′, X ′ ′ and Y are as defined above;
e 1) splitting off the protective group S 1 from compounds of the formula IX by known processes (for example Greene, Wuts, Protective Groups in Organic Synthesis, J. Wiley & Sons, New York 1991, Sonveaux, Bioorganic Chemistry 14 (1986) 274ff), for example, the monomethoxytrityl protective group is split off by treatment with acid, for example by treatment with 80% acetic acid, with 1-4% dichloroacetic acid in methylene chloride or chloroform, with 2% p-toluenesulfonic acid in DCM / methanol or by treatment with 1% trifluoroacetic acid in chloroform,
whereby compounds of formula XI are obtained

wherein
A, B ^PR , D, G, L, R⁵, R⁶, S², S³, X, X ′, X ′ ′ and Y are as defined above;
f₁) Compounds of the formula XI with compounds of the formula X according to the "phosphotriester method" known from oligonucleotide chemistry (Sonveaux, Bioorganic Chemistry 14 (1986) 274ff, Reese, J. Chem. Soc. Perkin Trans. 1993, 2291ff) in a suitable organic solvent, such as acetonitrile, benzene, DMF, DMSO, DCM, EE, chloroform, diethyl ether, tetramethyl urea, THF, N-methylpyrrolidone, petroleum ether, xylene or toluene or mixtures of suitable solvents, preferably in pyridine, at temperatures of -20 ° C to 100 ° C, preferably at 0 to 50 ° C, with the addition of a coupling reagent, such as 6-nitrobenzotriazol-1-yloxytris (dimethylamino) phosphonium hexafluorophosphate (NBOP, Hashmi, Nucleosides & Nucleotides 13 (1994) 1059), Benzotriazol-1-yloxy-tris (dimethylamino) phosphonium hexafluorophosphate (BOP, B. Castro, JR Dormoy, G. Evin and C. Selve, Tetrahedron Lett. 1975, 1219-1222), Benzotriazol-1-yloxy-tripyrrolidino-phosphonium hexafluorophosphate ( PyBOP, J. Coste, D. Le-Nguyen and B. Castro, Tetrahedron Lett. 1990, 205-208), O- (7-aza) benzotriazol-1-yltetramethyluronium hexafluorophosphate (HATU, L. Carpino, J. Am. Chem. Soc. 1993, 115, 4397), N, N-Bis [2-oxo -3-oxazolidinyl] phosphorodiamide chloride (Katti, Tetrahedron Lett. 26 (1985) 2547), 2-chloro-5,5-dimethyl-2-oxo-1,3,2-dioxaphosphorinane (Stawinski, Nucl. Acids Res., Symp 24, 1991, 229) or a compound of the formula XII

wherein
R¹⁵ for (C₆-C₁₂) aryl, optionally monosubstituted to tetrasubstituted by (C₁-C₆) alkyl, (C₁-C₆) alkoxy, nitro, chlorine, bromine and optionally one to 3 C atoms by heteroatoms, are preferred Nitrogen are substituted, ie for example for phenyl, tolyl, 2,4,6-trimethylphenyl, 2,4,6-tri isopropylphenyl, 2,3,5,6-tetramethylbenzene (Losse, Liebigs Ann. Chem. 1989, 19ff), 4-bromobenzene, 2-nitrobenzene, 4-nitrobenzene, 8-quinolyl, preferably 2,4,6-trimethylphenyl or 2,4,6-triisopropylphenyl, and
R¹⁶ represents a leaving group such as chlorine, bromine, imidazole, triazole, 4-nitroimidazole, 1,2,3,4-tetrazole, 3-nitro-1,2,4-triazole,
preferably with the addition of a coupling reagent of the compound of the formula XII or BOP, PyBOP or HATU,
optionally with the addition of a catalyst (Reese, J. Chem. Soc. Perkin Trans. 1993, 2291ff), such as, for example, N-methylimidazole, pyridine-N-oxides, such as 4-methoxy-pyridine-N-oxide or 4-ethoxy-pyridine -N-oxide, 4,6-dinitro-1-hydroxybenzotriazole, 1-hydroxy-5-phenyltetrazole, 1-hydroxy- (5- (4-nitrophenyl) tetrazole, 3-nitro-1H-1,2,4- triazol, 5- (3-nitrophenyl) -1H-tetrazole, 5- (3,5-dinitrophenyl) -1H-tetrazole, 5 - (- 1-methylimidazol-2-yl) -1H-tetrazole, 5 - [(1 -methylimidazol-2-yl) methyl] -1H-tetrazole or 1-hydroxy-4-nitro- 6- (trifluoromethyl) benzotriazole, preferably with 4-ethoxypyridine-N-oxide or 4-methoxy-pyridine-N-oxide as catalyst ,
the coupling reagents can be prepared in situ, or else separately and the solution of the activated species can be added in a suitable solvent,
to compounds of formula XIII

wherein
A, B ^PR , D, G, L, R⁵, R⁶, S¹, S², S³, X, X ′, X ′ ′ and Y are as defined above;
g₁) starting from compounds of the formula XIII, the steps e₁) and f₁) are repeated to the desired chain length, resulting in compounds of the formula XIV,

wherein
A, B ^PR , D, G, L, R⁵, R⁶, S¹, S², S³, X, X ′, X ′ ′, Y and n are as defined above;
h₁) the protective groups S¹, S², and S³ and the protective groups at B ^{PR are} split off according to known processes (e.g. Greene, Wuts, Protective Groups in Organic Synthesis, J. Wiley & Sons, New York 1991), ie for example the protective group S¹ as described in step e₁), the protecting groups S² or S³, if they stand for 2- (4-nitrophenyl) ethyl, by treatment with 0.5M 1,8-diazabicyclo [5,4,0] undec-7-en ( DBU) in pyridine or acetonitrile at room temperature, if S² or S³ are phenyl, by treatment with aqueous ammonia, if S² or S³ are allyl, by treatment with Pd [P (C₆H₅) ₃] ₄ and triphenylphosphine in DCM (Hayakawa et al., J. Org. Chem. 58 (1993) 5551) if S² or S³ is 2-cyanoethyl, by treatment with triethylamine in pyridine, or if S² or S³ is 2,2,2-trichloro-1,1 -dimethylethyl, by treatment with tributylphosphine, and the protective groups on B ^PR , for example if R¹⁴ is para-nitrophenylethyloxycar bonyl, with 0.5M DBU in pyridine, if R¹⁴ is isobutyryl or benzoyl or para- (methoxy) benzoyl, with conc. NH₄OH at 20 to 60 ° C or, if R¹³ is 2- (4-nitrophenyl) ethyl, by treatment with 0.5M DBU in pyridine or acetonitrile, is preferred if S¹ is monomethoxytrityl and S² is 2- (para-nitrophenyl) ethyl is preferably first cleaved off the monomethoxytrityl group as described in e₁), then cleaving S² as described, then the remaining protective groups, for example on the nucleobases, removed;
and optionally introduces groups Q and Q 'according to methods known to those skilled in the art (see, for example, Uhlmann & Peyman, Chem. Rev. 90 (1990) 543; M. Manoharan in "Antisense Research and Applications", Crooke and Lebleu, Eds., CRC Press, Boca Raton, 1993, Chapter 17, pp. 303ff; EP-A 0 552 766; S. Agrawal in Methods in Molecular Biology, Vol. 26, pp. 93ff, Humana Press, Totowa 1994), and optionally those obtained Connections according to Wang, Nucl. Acids Res. 22 (1994) 2326 cyclized, resulting in compounds of formula I.

Alternatively, conjugates Q 'can also be known to those skilled in the art Methods (J. March, "Advanced Organic Chemistry", Fourth Ed., J. Wiley & Sons, 1992) in the monomer blocks of formula XXII, which then according to the processes mentioned in the compounds of formula I. to be built in.

Compounds of formula XXII can be used, for example, for Q ′ = alkyl produce by reacting compounds of formula XXIII with Compounds of the formulas VIa or VIb and other reactions analogous to it was described for the formulas Va and Vb.

Compounds of formula XXII can also be prepared from compounds of Formula IX by splitting off the protective group S¹ and introducing the group Q ′ by known methods (J. March, "Advanced Organic Chemistry", Fourth Ed., J. Wiley & Sons, 1992).

Alternatively, conjugates Q and Q '' can also be known to those skilled in the art Methods (J. March, "Advanced Organic Chemistry", Fourth Ed., J. Wiley & Sons, 1992) in the monomer blocks of Formula XXIV, which then according to the processes mentioned in the compounds of formula I. to be built in.

Coupling reagents for the formation of peptide bonds (see c₁)) described for example in Houben-Weyl, Methods of Organic Chemistry, Volume 15/2, Georg Thieme Verlag Stuttgart 1974 and other reagents such as e.g. B. BOP (B. Castro, J.R. Dormoy, G. Evin and C. Selve, Tetrahedron Lett. 1975, 1219-1222), PyBOP (J. Coste, D. Le-Nguyen and B. Castro, Tetrahedron Lett. 1990, 205-208), BroP (J. Coste, M.-N. Dufour, A. Pantaloni and B. Castro, Tetrahedron Lett. 1990, 669-672), PyBroP (J. Coste, E. Frerot, P. Jouin and B. Castro, Tetrahedron Lett. 1991, 1967-1970) and uronium reagents such as e.g. B. HBTU (V. Dourtoglou, B. Gross, V. Lambropoulou, C. Zioudrou, Synthesis 1984, 572-574), TBTU, TPTU, TSTU, TNTU (R. Knorr, A. Trzeciak, W. Bannwarth and D. Gillessen, Tetrahedron Letters 1989, 1927-1930), TOTU (EP-A-0 460 446), HATU (L.A. Carpino, J. Am. Chem. Soc. 1993, 115, 4397-4398), HAPyU,  TAPipU (A. Ehrlich, S. Rothemund, M. Brudel, M. Beyermann, L.A. Carpino and M. Bienert, Tetrahedron Lett. 1993, 4781-4784), BOI (K. Akaji, N. Kuriyama, T. Kimura, Y. Fujiwara and Y. Kiso, Tetrahedron Lett. 1992, 3177-3180) or Acid chlorides or acid fluorides (L.A. Carpino, H.G. Chao, M. Beyermann and M. Bienert, J. Org. Chem., 56 (1991), 2635; J.-N. Bertho, A. Loffet, C. Pinel, F. Reuther and G. Sennyey in E. Giralt and D. Andreu (Eds.) Peptides 1990, Escom Science Publishers B. V.1991, pp. 53-54; J. Green and K. Bradley, Tetrahedron 1993, 4141-4146), 2,4,6-mesitylenesulfonyl-3-nitro-1,2,4-triazolide (MSNT) (B. Blankemeyer-crowd, M. Nimitz and R. Frank, Tetrahedron Lett. 1990, 1701-1704), 2,5-Diphenyl-2,3-dihydro-3-oxo-4-hydroxythiophene dioxide (TDO) (R. Kirstgen, R.C. Sheppard, W. Steglich, J. Chem. Soc. Chem. Commun. 1987, 1870-1871) or activated esters (D. Hudson, Peptide Res. 1990, 51-55) in the respective references.

Also preferred is the use of carbodiimides, e.g. B. Dicyclohexylcarbodiimide or diisopropylcarbodiimide. Preferably used are also phosphonium reagents, such as. B. PyBOP or PyBroP, Uronium reagents such as B. HBTU, TBTU, TPTU, TSTU, TNTU, TOTU or HATU, BOI.

The coupling can be carried out directly by adding compounds of the formula VIII with the activation reagent and optionally with the addition of additives such as e.g. B. 1-Hydroxybenzotriazole (HOBt) (W. Koenig, R. Geiger, Chem. Ber. 103, 788 (1970)) or 3-hydroxy-4-oxo-3,4-dihydrobenzotriazine (HOObt) (W. König, R. Geiger, Chem. Ber. 103, 2034 (1970)) or the Pre-activation of the building block as an activated ester can be done separately and the Solution of the activated species in a suitable solvent is added will.

The invention further relates to a method for producing the Compounds of formula I, wherein n is 1 to 100, thereby is characterized in that in the compounds of the formulas XV and XVI,

wherein
A, B ^PR , D, G, L, R⁵, R⁶, S¹, S², S³, X, X ′, X ′ ′ and Y are as defined above, o and p independently of one another zero to 50, preferably zero to 20 and o + p + 1 = n mean;
a₂) splits off the protective group S¹ in the compounds of the formula XV as described under e₁),
b₂) in the compounds of formula XVI splits off the protective group S³ as described under d₁) and
c₂) couples the resulting compounds to one another as described under f₁), resulting in compounds of the formula XIV,

wherein
A, B ^PR , D, G, L, R⁵, R⁶, S¹, S², S³, X, X ′, X ′ ′, Y and n are as defined above,
d₂) and this as described under h₁) to compounds of formula I.

The invention further relates to a process for the preparation of the compounds of the formula I, which is characterized in that
a₃) compounds of the formula X,

wherein
A, B ^PR , D, G, L, R⁵, R⁶, S¹, S², X, X ′, X ′ ′ and Y are as defined above,
coupled to a solid support via a SPACER using known processes, to give compounds of the formula XVII,

wherein
A, B ^PR , D, G, L, R⁵, R⁶, S¹, S², X, X ′, X ′ ′ and Y are as defined above,
SS stands for a solid support suitable for solid phase synthesis, such as, for example, aminopropyl-CPG (CPG = ®Controlled Pore Glass) or ®Tentagel, and
SPACER stands for a group which can be split off from the support after synthesis, as is known to the person skilled in the art (Sonveaux, Bioorganic Chemistry 14 (1986) 274ff), for example the bis (hydroxyethyl) sulfonyl group as described in EP-A 0 552 766 (HOE 92 / F012), or SPACER for bisfunctional conjugate molecules Q which are linked to the solid support via known cleavable groups, for example for nucleotides or oligonucleotides which are bound to the solid support via a succinic acid residue (Sonveaux, Bioorganic Chemistry 14 (1986) 274ff) or poly- or oligoethylene glycols, which are bound to the solid support via a succinic acid residue (Jäschke, Tetrahedron Lett. 34 (1993) 301) or, for example, cholesterol derivatives, which are bound to the solid support via a succinic acid residue (MacKellar, Nucl. Acids Res. 20 (1992) 3411);
b₃) from compounds of the formula XVII,

wherein
A, B ^PR , D, G, L, R⁵, R⁶, S¹, S², SS, SPACER, X, X ′, X ′ ′ and Y are as defined above,
splits off the protective group S¹ as described under e₁);
c₃) the resulting compound with compounds of the formula X,

wherein
A, B ^PR , D, G, L, R⁵, R⁶, S¹, S², X ′, X ′ ′ and Y are as defined above, as described under f₁);
d₃) steps b₃) and c₃) are repeated until the desired chain length;
e₃) optionally conjugates Q ′ by known methods (see, for example, Uhlmann & Peyman, Chem. Rev. 90 (1990) 543; M. Manoharar in "Antisense Research and Applications", Crooke and Lebleu, Eds., CRC Press, Boca Raton , 1993, Chapter 17, pp. 303ff; EP-A 0 552 766; S. Agrawal in "Methods in Molecular Biology", Vol. 26, pp. 93ff, Humana Press, Totowa 1994);
f₃) the compounds produced in this way are split off from the solid support by known processes, for example the bis (hydroxyethyl) sulfonyl linker, as described in EP-A 0 552 766 (HOE92 / F012), by treatment with DBU, the succinic acid linker by treatment with aqueous ammonia and the protecting groups as described in step h 1), it being possible for the protecting groups to be cleaved off from the support before the cleavage.

The compounds of formula I find inhibitors of gene expression Use. The invention therefore relates to the use of therapeutically active compounds according to the invention for the preparation of a Medicament and a method for producing a medicament, the is characterized in that the compounds of the invention with a physiologically acceptable carrier and, if appropriate, suitable ones Additives and / or auxiliaries mixed.

Therapeutically active compounds are generally understood to be those those due to the sequence of building blocks B that correspond to the nucleobases Act as analogues of antisense oligonucleotides, triple helix forming oligonucleotides, aptamers (RNA or DNA molecules that specific target molecules, e.g. B. proteins or receptors, can bind (z. B. L.C. Bock et al., Nature 1992, 355, 564) or ribozymes (catalytic RNA, see. e.g. B. Castanetto et al., Critical Rev. Eukar. Gene Expr. 1992, 2, 331), especially as analogs of antisense oligonucleotides and triple helix forming oligonucleotides.

In addition, another object of the present invention is the Use of the compounds according to the invention as a diagnostic agent, for example to detect the presence or absence or the amount of one specific double-stranded or single-stranded nucleic acid molecule in a biological sample.  

The compounds according to the invention have for the invention Use a length (n-1) of approximately 6-100, preferably approximately 10-40, especially from about 12-31 nucleotides. Otherwise, the above also apply here Preferred areas, modifications or conjugations described.

For example, the pharmaceutical compositions of the present invention can be used for Treatment of diseases caused by viruses, for example through HIV, HSV-1, HSV-2, influenza, VSV, hepatitis B or Papilloma viruses.

Sequences according to the invention (base sequences) which act against such targets are effective, for example:

• a) against HIV, e.g. B.
• b) against HSV-1, e.g. B.

For example, the pharmaceutical compositions of the present invention are also suitable for Treatment of cancer or restenosis. For example, Sequences (base sequences) are used that are directed against targets are responsible for the development or growth of cancer. Such Examples of targets are:

• 1) Nuclear oncoproteins such as c-myc, N-myc, c-myb, c-fos, c-fos / jun, PCNA, p120
• 2) cytoplasmic / membrane-associated oncoproteins such as EJ-ras, c-Ha-ras, N-ras, rrg, bcl-2, cdc-2, c-raf-1, c-mos, c-src, c-abl
• 3) Cellular receptors such as EG F receptor, c-erbA, retinoid Receptors, protein kinase regulatory subunit, c-fms
• 4) cytokines, growth factors, extracellular matrix such as CSF-1, IL-6, IL-1a, IL-Ib, IL-2, IL-4, bFGF, myeloblastin, fibronectin.

Sequences according to the invention (base sequences) which act against such targets are effective, for example

• a) against c-Ha-ras, e.g. B.
• c) c-myc, e.g. B.
• d) c-myb, e.g. B.
• e) c-fos, e.g. B.
• f) p120, e.g. B.
• g) EGF receptor, e.g. B.
• h) p53 tumor suppressor, e.g. B.
• i) bFGF, e.g. B.

The drugs of the present invention are also useful, for example Treatment of diseases caused by integrins or cell-cell Adhesion receptors are influenced, for example by VLA-4, VLA-2, ICAM or ELAM.

Sequences according to the invention (base sequences) which act against such targets are effective, for example

• a) VLA-4, e.g. B.
• b) ICAM, e.g. B.
• c) ELAM-1, e.g. B.

The drugs of the present invention are also useful, for example Treatment of diseases triggered by factors such as TNF alpha will.

Sequences according to the invention (base sequences) which act against such targets are effective, for example

• a) TNF-alpha, e.g. B.

The drugs can e.g. B. in the form of pharmaceutical preparations that one for example topically or orally, e.g. B. in the form of tablets, coated tablets, hard or Administer soft gelatin capsules, solutions, emulsions or suspensions can be used. You can also rectally z. B. in the form of suppositories or parenterally z. B. in the form of solutions for injection. For the Production of pharmaceutical preparations can include these compounds in processed therapeutically inert organic and inorganic carriers will. Examples of such carriers for tablets, coated tablets and Hard gelatin capsules are lactose, corn starch or derivatives thereof, talc and Stearic acid or salts thereof. Suitable carriers for the production of Solutions are water, polyols, sucrose, invert sugar and glucose. Suitable carriers for injection solutions are water, alcohols, polyols, glycerol and vegetable oils. Suitable carriers for suppositories are vegetable and hardened oils, waxes, fats and semi-liquid polyols. The pharmaceutical Preparations can also contain preservatives, solvents, stabilizers, Wetting agents, emulsifiers, sweeteners, colorants, flavoring agents, salts for Change in osmotic pressure, buffers, coating agents, antioxidants, as well as possibly contain other therapeutic agents.

A preferred application is oral application. Another preferred The form of administration is injection. For this, the antisense Oligonucleotides in a liquid solution, preferably in a physiological one acceptable buffers, e.g. B. Hank's solution or Ringer's solution. The therapeutically active compounds according to the invention can, however can also be formulated in a solid form and solved before use or be suspended. The preferred for systematic administration Dosages are approximately 0.01 mg / kg to approximately 50 mg / kg body weight and day.  

Sequence list:

ACACCCAATTCTGAAAATGG (I)
AGGTCCCTGTTCGGGCGCCA (II)
GTCGACACCCAATTCTGAAAATGGATAA (III)
GCTATGTCGACACCCAATTCTGAAA (IV)
GTCGCTGTCTCCGCTTCTTCTTCCTG (V)
GTCTCCGCTTCTTCTTCCTGCCATAGG (VI)
GCGGGGCTCCATGGGGGTCG (VII)
CAGCT GCAACCCAGC (VIII)
GGCTGCTGGAGCGGGGCACAC (IX)
AACGTTGAGGGGCAT (X)
GTGCCGGGGTCTTCGGGC (XI)
GGAGAACATCATGGTCGAAAG (XII)
CCCGAGAACATCATGGTCGAAG (XIII)
GGGGAAAGCCCGGCAAGGGG (XIV)
CACCCGCCTTGGCCTCCCAC (XV)
GGGACTCCGGCGCAGCGC (XVI)
G GCAAACTTTCTTTTCCTCC (XVII)
GGGAAGGAGGAGGATGAGG (XVIII)
GGCAGTCATCCAGCTTCGGAG (XIX)
GCAGTAAGCATCCATATC (XX)
CCCCCACCACTTCCCCTCTC (XXI)
CTCCCCCACCACTTCCCCTC (XXII)
GCTGGGAGCCATAGCGAGG (XXIII)
ACTGCTGCCTCTTGTCTCAGG (XXIV)
CAATCAATG ACTTCAAGAGTTC (XXV)
GGTCCCTGTTCGGGCGCCA (XXVI)
GTGCCGGGGTCTTCGGG (XXVII)
GGAGGATGCTGAGGAGG (XXVIII)
GGAGGATGCTGAGG (XXIX)
CAGGAGGATGCTGAGGAGG (XXX)
GGCTGCCATGGTCCC (XXXI)
TCATGGTGTCCTTTGCAGCC (XXXII)
TCATGGTGTCCTTTGCAG (XXXIII).

Examples 1) N- (4-methoxytriphenylmethoxy) ethylaminomethanephosphonic acid di (2- (p-ni- trophenyl) ethyl) ester 1a) N-fluorenylmethyloxycarbonyl-2-aminoethanol

8.61 g (0.141 mol) of 2-aminoethanol were in 250 ml of dioxane and 150 ml of H₂O solved. At 15-20 ° C were first 17.79 g (0.212 mol) NaHCO₃, then in portions 50 g (0.148 mol) fluorenylmethyloxycarbonyl-N-succinimide admitted. The mixture was stirred at room temperature for 1 h, then became dry evaporated. The residue was between dichloromethane (DCM) and H₂O distributed, the organic phase dried over Na₂SO₄ and the solvent in Vacuum evaporated. The residue was stirred with 100 ml of ether Aspirated product and washed well with ether. The yield was 38.77 g (97%).

MS (ES +) 284.2 (M + H) ⁺; 1 H-NMR (200 MHz, DMSO, TMS): δ = 3.05 (dd, 2H, CH₂OH); 3.39 (dd, 2H, N-CH₂); 4.25 (m, 3H, Ar-CH-CH₂); 4.61 (t, 1H, OH); 7.14-7.98 (m, 15H, Ar-H, NH).

1b) N-Fluorenylmethyloxycarbonyl-2-amino-1- (4-methoxytriphenylmethoxy) et-han

10 g (35.3 mmol) of N-fluorenylmethyloxycarbonyl-2-aminoethanol (from example  1a), dissolved in 100 ml absolute. N, N-Dimethylformamide (DMF) were at 0 ° C with 5.93 g (45.93 mmol) diisopropylethylamine (DIPEA) and 10.91 g (35.3 mmol) 4-methoxytriphenylmethyl chloride was added and first at 0 ° C. for 1 h, then at 1 h Room temperature stirred. The reaction mixture was evaporated and distributed between DCM and a saturated aqueous NaHCO₃ solution. The organic phase was washed with H₂O, dried over Na₂SO₄ and that Evaporated solvent in vacuo. For cleaning was over silica gel chromatographed (first n-heptane / ethyl acetate (EE) / triethylamine (TEA) 70/29/1; then EE / TEA 99/1). The yield was 14.4 g (73%).

MS (FAB): 562.3 (M + Li) ⁺; 1 H-NMR (200 MHz, DMSO, TMS): δ = 2.95 (t, 2H, CH₂O-MMTr); 3.21 (dd, 2H, N-CH₂); 3.75 (s, 3H, OCH3); 4.25 (m, 3H, Ar-CH- CH₂); 4.61 (t, 1H, OH); 6.80-7.96 (m, 23H, Ar-H, NH).

1c) 2-Amino-1- (4-methoxytriphenylmethoxy) ethane

5.0 g (9 mmol) N-fluorenylmethyloxycarbonyl-2-amino-1- (4-methoxy triphenylmethoxy) ethane (from Example 1b), dissolved in 50 ml of absolute. DMF were added at room temperature with 6.55 g (90 mmol) of diethylamine and 2 h touched. For purification, it was chromatographed on silica gel (first n-heptane / EE / TEA 50/49/1; then EE / methanol / TEA 79/20/1). The yield was 2.96 g (98.7%).

MS (ES +): 340.3 (M + Li) ⁺; 1 H-NMR (200 MHz, DMSO, TMS): δ = 2.75 (t, 2H, CH₂O-MMTr); 2.93 (dd, 2H, N-CH₂); 3.75 (s, 3H, OCH3); 6.83-7.47 (m, 14H, Ar-H).

1d) 2-methylimino-1- (4-methoxytriphenylmethoxy) ethane (trimer)

2.96 g (8.9 mmol) of 2-amino-1- (4-methoxytriphenylmethoxy) ethane (from example 1c), dissolved in 10 ml of methanol, were cooled with ice with 1.08 g (13.22 mmol)  37% formaldehyde were added and the mixture was stirred at room temperature for 4 h, during which a viscous precipitation formed. The reaction mixture was evaporated and the Purification chromatographed on silica gel (n-heptane / EE / TEA 50/49/1). The Yield was 1.7 g (55%).

MS (FAB) 1042.8 (M + Li) ⁺; 1034.8 (M-H) ⁺. 1 H-NMR (200MHz, DMSO, TMS): δ = 2.60 (t, 6H, O-CH₂); 2.99 (t, 6H, N-CH₂); 3.69 (s, 9H, OCH₃); 6.78-7.42 (m, 42H, Ar-H).

1e) Di (2- (4-nitrophenyl) ethyl) phosphite

23.42 g (0.1 mol) of diphenyl phosphite together with 33.43 g (0.2 mol) p-Nitrophenylethanol heated under argon at 100 ° C for 14 h, for cleaning Chromatographed on silica gel (n-heptane / EE 50/50; then EE / methanol 80/20). Yield: 55%.

MS (FAB) 403.1 (M + Na) ⁺; 381.1 (M + H) ⁺. 1 H-NMR (200MHz, DMSO, TMS): δ = 3.03 (t, 4H, Ar-CH₂); 4.20 (4H, dt, O-CH₂); 6.71 (d, J = 140Hz; 1H, PH); 7.52 (d, 4H, Ar-H); 8.17 (d, 4H, Ar-H).

1f) N- (4-methoxytriphenylmethoxy) ethylaminomethanephosphonic acid di (2- (p-nitro phenyl) ethyl) ester

500 mg (1.32 mmol) of di (2- (4-nitrophenyl) ethyl) phosphite (from Example 1e), dissolved in 2 ml absolute. Tetrahydrofuran (THF) was 341 mg (0.329 mmol) 2-methylimino-1- (4-methoxytriphenylmethoxy) ethane (trimer) (from Example 1d) added and the mixture was stirred at 80 ° C for 3 h. The solvent was evaporated and the mixture was stirred at 100 ° C. for a further 30 min. Was for cleaning Chromatographed on silica gel (first EE / TEA 99/1; then EE / methanol / TEA 90/9/1). Yield: 83%.  

MS (FAB) 732.3 (M + Li) ⁺. 1 H-NMR (200MHz, DMSO, TMS): δ = 2.64-3.06 (m, 10H, Ar-CH₂ + P-CH₂ + CH₂-OMMTr + N-CH₂); 3.73 (s, 3H, OCH₃); 4.16 (German, 4H, PO-CH₂); 6.78-8.08 (m, 22H, Ar-H).

2) N- (N⁶-anisoyl) cytosin-1-yl-acetyl-N- (4-methoxytriphenylmethoxy) ethyl-amino methanephosphonic acid di (2- (p-nitrophenyl) ethyl) ester

2a) To 2.00 g (2.76 mmol) of N- (4-methoxytriphenylmethoxy) ethylamino methanephosphonic acid di (2- (p-nitrophenyl) ethyl) ester (from Example 1f), dissolved in 60 ml absolute. DMF, 0.952 g (8.27 mmol) of N-ethylmorpholine (NEM), 0.834 g (2.76 mmol) (N⁶-anisoyl) cytosin-1-yl-acetic acid and 1.153 g (3.03 mmol) O- (7-aza) benzotriazol-1-yltetramethyluronium hexafluorophosphate (HATU, L. Carpino, J. Am. Chem. Soc. 1993, 115, 4397) added and 12 h at Room temperature stirred. After that, the same amount of HATU again added and a further 3 h at room temp. touched. For cleaning was on Chromatographed silica gel (DCM / methanol / TEA 95/4/1). The yield was 2.7 g (97%).

MS (ES +): 1012.0 (M + H) ⁺. 1 H-NMR (200MHz, DMSO, TMS): Δ = 2.94 (t, 4H, P-O-CH₂-CH₂-Ar); 3.06 (t, 2H, MMTr-O-CH₂); 3.23-3.63 (m, 4H, P-CH₂ + N- CH₂); 3.75 (s, 3H, OCH₃); 3.83 (s, 3H, OCH₃); 4.10 (dt, 4H, P-O-CH₂); 4.79 (s, broad, 2H, CO-CH₂); 6.80-8.18 (m, 28H, Ar-H, cytosinyl-H); 11.03 (broad s, 1H, NH).

2b) Approach as in Example 2a, but using O- (cyan (ethoxycarbonyl) methylenamino) -1,1,3,3-tetramethyluronium tetrafluoroborate (TOTU, EP 0460446) instead of HATU. The yield was 57%. Spectroscopic data: s. Example 2a.  

3) N- (N⁶-anisoyl) cytosin-1-yl-acetyl-N- (4-methoxytriphenylmethoxy) ethyl-amino methanephosphonic acid (2- (p-nitrophenyl) ethyl) monoester (triethylammonium salt)

1 g (0.99 mmol) of N- (N⁶-anisoyl) cytosin-1-yl-acetyl-N- (4-methoxytriphenyl methoxy) ethylaminomethanephosphonic acid di (2- (p-nitrophenyl) ethyl) ester- (from Example 2) were in 20 ml of a 0.1M solution of 1,8-diazabicyclo [5.4.0] undec- 7s (DBU) in absolute. Acetonitrile dissolved and 4 h at room temperature. touched. The Reaction mixture was between DCM and an aqueous KH₂PO₄ solution (pH 7), the organic phase was dried over Na₂SO₄ and that Evaporated solvent in vacuo. For cleaning was over silica gel chromatographed (EA / methanol / TEA 70/29/1). The yield was 540 mg (57%).

MS (FAB): 906.5 (M-H + 2Na) ⁺; 884.6 (M + Na) ⁺; 862.5 (M + H) ⁺. 1 H NMR (200MHz, DMSO, TMS): δ = 3.00 (m, 4H, P-O-CH₂-CH₂-Ar + MMTr-O-CH₂); 3.38-3.60 (m, 4H, P-CH₂ + N-CH₂); 3.73 (s, 3H, OCH₃); 3.82 (s, 3H, OCH₃); 4.01 (dt, 2H, P-O-CH₂); 4.79 & 5.03 (each s, broad, 2H, CO-CH₂); 6.78-8.20 (m, 24H, Ar-H, cytosinyl-H); 11.00 (broad s, 1H, NH).

4) N- (N⁶-anisoyl) cytosin-1-yl-acetyl-N- (2-hydroxy) ethylaminomethanephosph-hon acid di (2- (p-nitrophenyl) ethyl) ester

1.00 g (0.99 mmol) N- (N⁶-anisoyl) cytosin-1-yl-acetyl- N- (4-methoxytriphenylmethoxy) ethylaminomethanephosphonic acid di (2- (p-nitro phenyl) ethyl) esters (from Example 2) were 80% more aqueous in 80 ml Acetic acid dissolved and stirred for 4 h at room temperature. The solvent was evaporated and coevaporated twice with toluene. Was for cleaning Chromatographed on silica gel (EA / methanol / TEA 85/14/1). The yield was 522 mg (71%).  

MS (FAB): 761.2 (M + Na) ⁺; 739.3 (M + H) ⁺. 1 H-NMR (200MHz, DMSO, TMS): δ = 2.98 (t, 4H, P-O-CH₂-CH₂-Ar); 3.38-3.67 (m, 4H, N-CH₂CH₂-OH); 3.80-3.89 (m, 2H, P-CH₂); 3.91 (s, 3H, OCH₃); 4.12 (dt, 4H, P-O-CH₂); 4.78 & 4.87 (jew. s, broad, 2H, CO-CH₂); 6.98-8.19 (m, 14H, Ar-H, cytosinyl-H); 11.02 (see broad, 1H, NH).

5) 5′-MMTr-CA ^to -P (ONPE) -CA ^to -P (ONPE) ₂

The synthesis was carried out analogously to Example 17 from N- (N⁶-anisoyl) cytosin-1-yl-acetyl N- (4-methoxytriphenylmethoxy) ethylaminomethanephosphonic acid (2- (p-nitr-o phenyl) ethyl) monoester (triethylammonium salt) (Example 3) and N- (N⁶- Anisoyl) cytosin-1-yl-acetyl-N- (2-hydroxy) ethylaminomethanephosphonic acid redi (2- (p-nitro phenyl) ethyl) ester (Example 4). For cleaning was over silica gel chromatographed (EA / methanol / TEA 85/14/1). Yield: 73%.

MS (FAB) 1605 (M + Na) ⁺; 1583 (M + H) ⁺; 1 H-NMR (200MHz, DMSO, TMS): δ = 2.94-3.18 (m, 6H, P-O-CH₂-CH₂-Ar); 3.26-3.95 (m, 10H); 3.75 (s, 3H, OCH₃); 3.85 (s, 6H, OCH₃); 3.99-4.36 (m, 8H, P-O-CH₂); 4.75-4.92 (m, broad, 4H, CO- CH₂); 6.83-8.18 (m, 38H, Ar-H, cytosinyl-H); 10.98 & 11.03 (each broad, 2H, NH).

6) 5'-HO-C ^to -P (ONPE) -C ^to -P (ONPE) ₂

The synthesis was carried out analogously to Example 4 from "5'-MMTr-C ^An -P (ONPE) -C ^An - P (ONPE) ₂" (Example 5). For purification, it was chromatographed on silica gel (EA / methanol / TEA 85/14/1). The yield was 74%.

MS (FAB) 1332.4 (M + Na) ⁺; 1310.3 (M + H) ⁺;  

7) N- (4-methoxytriphenylmethoxy) ethylaminomethanephosphonic acid diethyl ester

The synthesis was carried out analogously to Example 1f, but with diethyl phosphite. Yield: 87.5%.

MS (FAB) 490.2 (M + Li) ⁺. 1 H-NMR (200 MHz, DMSO, TMS): δ = 1.22 (t, 6H, CH₂-CH₃); 2.80 (t, 2H, N-CH₂); 2.91 (d, J = 12.5Hz, 2H, P-CH₂); 3.02 (t, 2H, CH₂-OMMTr); 3.75 (s, 3H, OCH₃); 4.01 (dq, 4H, PO-CH₂); 6.84-7.45 (m, 14H, Ar-H).

8) N-Thymin-1-yl-acetyl-N- (4-methoxytriphenylmethoxy) ethylamino diethyl methanephosphonate

To 2.04 g (4.22 mmol) of N- (4-methoxytriphenylmethoxy) ethylaminomethane diethyl phosphonate (Example 7), dissolved in 50 ml of absolute. Were DMF added 570.3 mg (4.22 mmol) of hydroxybenzotriazole (HOBT), 972.1 mg (8.44 mmol) NEM, 777 mg (4.22 mmol) tymidin-1-yl-acetic acid and 639 mg (5.06 mmol) diisopropyl carbodiimide. The mixture was stirred at room temperature for 16 h Evaporated solvent, the residue was dissolved in DCM and with saturated aqueous NaHCO₃ solution, then with saturated aqueous NaCl Solution extracted. It was dried over sodium sulfate, the solvent evaporated. For purification, it was chromatographed on silica gel (EE / methanol / TEA 98/2/1). The yield was 2.47 g (90%).

MS (FAB): 662.3 (M + Na) ⁺; 656.3 (M + Li) ⁺. 1 H-NMR (200MHz, DMSO, TMS): δ = 1.12-1.32 (m, 6H, CH₂-CH₃); 1.68 & 1.75 (each s, 3H, T-CH₃); 3.10-3.40 (m, 2H, CH₂-OMMTr); 3.53-3.70 (m, 4H, P-CH₂ + N-CH₂); 3.75 (s, 3H, OCH₃); 3.83-4.16 (m, 4H, PO-CH₂); 4.62 & 4.72 (each s, 2H, CO-CH₂); 6.83-7.42 (m, 15H, Ar-H, T-H); 11.28 (s, 1H, NH).  

9) N-Thymin-1-yl-acetyl-N- (4-methoxytriphenylmethoxy) ethylamino monoethyl methanephosphonate (triethylammonium salt)

811 mg (1.25 mmol) of N-thymin-1-yl-acetyl-N- (4-methoxytriphenylmethoxy) ethyl diethyl aminomethanephosphonate (Example 8) were dissolved in 3.75 ml of 1N NaOH suspended. The mixture was stirred at room temperature for 3 h and then at 50 ° C. for 6 h. The reaction mixture was concentrated in vacuo, the residue over Chromatographed silica gel (EA / methanol / TEA 100/10/10, then 100/40/10). The yield was 897 mg (99.5%).

MS (ES-): 620.4 (M-H) ⁻. 1 H-NMR (200 MHz, DMSO, TMS): δ = 1.18 (t, 9H, N- CH₂-CH₃); 1.68 & 1.74 (each s, 3H, T-CH₃); 2.96-3.08 (q, 6H, N-CH₂-CH₃); 3.35 (m, 2H, N-CH₂); 3.43-3.70 (d, J = 11Hz, 2H, P-CH₂); 3.63 (t, 2H, CH₂- OMMTr); 3.75 (s, 3H, OCH₃); 3.78 (dq, 2H, PO-CH₂); 4.60 & 4.86 (each s, 2H, CO-CH₂); 6.82-7.41 (m, 15H, Ar-H, T-H); 11.24 (s, 1H, NH).

10) N-Thymin-1-yl-acetyl-N- (2-hydroxy) ethylaminomethanephosphonic acid diethyl ester

The synthesis was carried out analogously to Example 4 from N-thymin-1-yl-acetyl-N- (4-methoxy triphenylmethoxy) ethylaminomethanephosphonic acid diethyl ester (example 8th). For purification, it was chromatographed on silica gel (EA / methanol 90/10). Yield: 80%.

MS (FAB): 400.1 (M + Na) ⁺; 378.1 (M + H) ⁺. 1 H-NMR (200MHz, DMSO, TMS): δ = 1.17-1.32 (m, 6H, CH₂-CH₂); 1.78 (s, 3H, T-CH₃); 3.40-3.69 (m, 4H, CH₂-OH + N-CH₂); 3.89 (d, J = 11Hz, 2H, P-CH₂); 3.92-4.19 (m, 4H, PO-CH₂); 4.70 (s, 2H, CO-CH₂); 4.98 (t, 1H, OH); 7.22 & 7.30 (each s., 1H, T-H); 11.25 (s, 1H, NH).  

11) N- (4-methoxytriphenylmethoxy) ethylaminomethanephosphonic acid diphenyl ester

The synthesis was carried out analogously to Example 1f, but with diethyl phosphite. Yield: 100%.

MS (FAB) 58.2 (M + Li) ⁺.

12) N-Thymin-1-yl-acetyl-N- (4-methoxytriphenylmethoxy) ethylamino monophenyl methanephosphonate (triethylammonium salt)

The synthesis was carried out analogously to Example 8l from N- (4-methoxytriphenylmethoxy) diphenyl ethylaminomethanephosphonate (Example 11) and tymidin-1-yl acetic acid. For purification, it was chromatographed on silica gel (EE / methanol / TEA / H₂O 90/10/5 / 0.5). Yield: 47%.

MS (FAB): 682.3 (M + 2Li-H) ⁺. 1 H-NMR (200MHz, DMSO, TMS): δ = 1.16 (t, 9H, N-CH₂-CH₃); 1.67 & 1.72 (each s, 3H, T-CH₃); 2.96-3.70 (m, 12H, N-CH₂- CH₃ + N-CH₂ + P-CH₂ + CH₂-OMMTr); 3.75 (s, 3H, OCH₃); 4.58 & 4.88 (each s, 2H, CO-CH₂); 6.74-7.46 (m, 20H, Ar-H, T-H); 11.23 (s, 1H, NH).

13) N-Thymin-1-yl-acetyl-N- (4-methoxytriphenylmethoxy) ethylamino phenyl- (4-nitrophenylethyl) diester of methanephosphonate

385.4 mg (0.5 mmol) of N-thymin-1-yl-acetyl-N- (4-methoxytriphenylmethoxy) ethylaminomethanephosphonic acid monophenyl ester (triethylammonium salt) (Example 12) and 92 mg (0.55 mmol) (4-nitrophenylethanol) were added three times absolutely Pyridine coevaporated, then in 15 ml absolute. Pyridine dissolved. At 0 ° C 403.4 mg (0.15 mmol) of 3-nitro-1- (p-toluenesulfonyl) -1H-1,2,4-triazole  (TSNT) added, then stirred for 16 h at 0-5 ° C, the pyridine in vacuo distilled off, the residue taken up in EA and successively with saturated aqueous NaHCO₃ solution, then washed with NaCl solution. For cleaning was chromatographed on silica gel (EE / TEA 100/2). The yield was 162 mg.

MS (FAB): 831.3 (M + 2Li-H) ⁺; (M + Li) ⁺.

14) N-Thymin-1-yl-acetyl-N- (4-methoxytriphenylmethoxy) ethylamino methanephosphonic acid di (2- (p-nitrophenyl) ethyl) ester

The synthesis was carried out analogously to Example 8 from N- (4-methoxytriphenylmethoxy) ethylaminometha 44552 00070 552 001000280000000200012000285914444100040 0002019543865 00004 44433nphosphonsäuredi (2- (p-nitrophenyl) ethyl) ester (Example 1f) and Tymidin-1-yl acetic acid. Yield: 63%

MS (ES +): 898.4 (M + Li) ⁺. 1 H-NMR (200MHz, DMSO, TMS): δ = 1.65 & 1.72 (jew. s, 3H, T-CH₃); 2.96 (t, 4H, P-O-CH₂-CH₂-Ar); 3.06 (t, 2H, N-CH₂); 3.67 (d, J = 11Hz, 2H, P-CH₂); 3.70 (m, 2H, MMTr-O-CH₂); 3.75 (s, 3H, OCH₃); 3.83 (s, 3H, OCH₃); 4.10 (dt, 4H, P-O-CH₂); 4.59 & 4.62 (each, broad, 2H, CO-CH₂); 6.83-8.18 (m, 23H, Ar-H, T-H); 11.30 (broad s, 1H, NH).

15) N-Thymin-1-yl-acetyl-N- (4-methoxytriphenylmethoxy) ethylamino methanephosphonic acid (4-nitrophenylethyl) monoester (triethylammonium salt) 15a) From 30 mg of N-thymin-1-yl-acetyl-N- (4-methoxytriphenylmethoxy) ethylaminomethanephosphonic acid phenyl- (4-nitrophenylethyl) diester (Example 13)

30 mg of N-thymin-1-yl-acetyl-N- (4-methoxytriphenylmethoxy) ethylamino  phenyl- (4-nitrophenylethyl) diesters of methanephosphonate (Example 13) were described in a mixture of 1 ml TEA, 1 ml dioxane and 80 mg p-nitrobenzaldoxime and stirred for 3 h at room temperature. The solvent was removed in vacuo evaporated, the residue three times with pyridine and twice with toluene coevaporated. The residue was chromatographed on silica gel (EA / TEA 100/2, then EE / methanol / TEA 60/40/2). The yield was 23 mg.

MS (FAB): 755.3 (M + 2Li-H) ⁺. 1 H-NMR (250 MHz, DMSO, TMS): δ = 1.15 (t, 9H, N-CH₂-CH₃); 1.60 & 1.79 (m, 3H, T-CH₃); 2.80-3.3.60 (m, 14H, N-CH₂- CH₃ + N-CH₂ + P-CH₂ + CH₂-OMMTr + Ar-CH₂); 3.73 (s, 3H, OCH₃); 4.01 (German, 2H, P-O-CH₂); 4.58-4.92 (m, 2H, CO-CH₂); 6.82-8.18 (m, 19H, Ar-H, T-H); 11.30 (s, 1H, NH).

15b) From N-thymin-1-yl-acetyl-N- (4-methoxytriphenylmethoxy) ethylamino methanephosphonic acid di (2- (p-nitrophenyl) ethyl) ester (Example 14)

The synthesis was carried out analogously to Example 3 from N-thymin-1-yl-acetyl N- (4-methoxytriphenylmethoxy) ethylaminomethanephosphonic acid di (2- (p-nitro phenyl) ethyl) ester (Example 14), but in pyridine as solvent. Yield: 82%. Spectoscopic data see Example 15a.

16) N-Thymin-1-yl-acetyl-N- (4-methoxytriphenylmethoxy) ethylaminomethane phosphonic acid

The synthesis was carried out analogously to Example 15b. As a by-product in 18% Yield N-thymin-1-yl-acetyl-N- (4-methoxytriphenylmethoxy) ethylamino Get methanephosphonic acid.

MS (ES-): 592.2 (M-H) ⁻.  

17) 5'-MMTr-T-P (OEthyl) -T-P (OEthyl) ₂

361 mg (0.5 mmol) of N-thymin-1-yl-acetyl-N- (4-methoxytriphenylmethoxy) ethylaminomethanephosphonic acid monoethyl ester (triethylammonium salt) (IX) and 188.7 mg (0.5 mmol) of N-thymin-1-yl-acetyl-N- (2-hydroxy) ethylamino diethyl methanephosphonate (X) were together twice with absolute. Pyridine coevaporated, then in 10 ml of absolute. Pyridine dissolved. At 5-10 ° C 1.5 mmol TSNT were added and it was 16 h at room temp. touched. The Pyridine was evaporated in vacuo, the residue was dissolved in EA and successively with saturated aqueous NaHCO₃ solution, then with NaCl solution washed. It was dried over Na₂SO₄, concentrated and for cleaning was chromatographed on silica gel (EA / methanol / TEA 92/8/2). The Yield was 223 mg (46%).

MS (FAB): 987.5 (M + Li) ⁺. 1 H-NMR (200MHz, DMSO, TMS) characteristic Signals are: Ar-H & Thymine-H: 6.82-7.43 (m, 16H); CO-CH₂: 4.59-4.78 (m, 4H); Thymine CH₃: 1.63-1.80 (m, 6H).

18) 5'-HO-T-P (OEthyl) -T-P (OEthyl) ₂

The synthesis was carried out analogously to Example 4 from 5'-MMTr-T-P (OEthyl) -T-P (OEthyl) ₂ (Example 17). For purification, it was chromatographed on silica gel (EE / methanol / TEA 85/15/2, then 100/50 / 1.5). The yield was 95%.

MS (FAB): 731.2 (M + Na) ⁺; 709.1 (M + H) ⁺. 1 H-NMR (200MHz, DMSO, TMS) Characteristic signals are: Thymine-H: 7.21-7.36 (m, 2H); CO-CH₂: 4.60- 4.76 (m, 4H); Thymine CH₃: 1.63-1.79 (m, 6H).  

19) 5'-MMTr-T-P (OPhenyl) -T-P (OEthyl) ₂

The synthesis was carried out analogously to Example 17 from N-thymin-1-yl-acetyl-N- (2-hy droxy) ethylaminomethanphosphonsäurediethylester (Example 10) and N-thymin-1-yl-acetyl-N- (4-methoxytriphenylmethoxy) ethylaminomethaneph-osphon acid monophenyl ester (triethylammonium salt) (Example 12). For cleaning was chromatographed on silica gel (EA / methanol / TEA 93/7/2). Yield: 58%.

MS (FAB): 1051.4 (M + Na) ⁺; 1029.5 (M + H) ⁺. 1 H-NMR (200MHz, DMSO, TMS) Characteristic signals are: Ar-H & Thymin-H: 6.82-7.53 (m, 21H); CO-CH₂: 4.52-4.82 (m, 4H); Thymine-CH₃: 1.62-1.80 (m, 6H).

20) N-Thymin-1-yl-acetyl-N- (2-hydroxyethyl) aminomethanephosphonic acid di (4-nitro phenylethyl) ester

The synthesis was carried out analogously to Example 4 from N-thymin-1-yl-acetyl-N- (4-methoxy triphenylmethoxy) ethylaminomethanephosphonic acid di (2- (p-nitro phenyl) ethyl) ester (Example 14). For cleaning was over silica gel chromatographed (EA / methanol 90/10). Yield: 85%.

MS (ES +): 620.3 (M + H) ⁺. 1 H-NMR (500 MHz, DMSO, TMS): δ = 1.73 (s, 3H, T-CH₃); 2.97 (t, 4H, P-O-CH₂-CH₂-Ar); 3.41 (m, 2H, N-CH₂); 3.59 (m, 2H, CH₂- OH); 3.83 (d, 2H, J = 11Hz; P-CH₂); 4.08-4.30 (m, 4H, P-O-CH₂); 4.54 & 4.78 (jew. s, broad, 2H, CO-CH₂); 4.99 (t, 1H, OH); 7.14-8.19 (m, 9H, Ar-H, Thymidinyl-H); 11.30 (broad s, 1H, NH).

21) 5'-MMTr-T-P (ONPE) -T-P (OEthyl) ₂

The synthesis was carried out analogously to Example 17 from N-thymin-1-yl-acetyl-N- (2-hy  droxy) ethylaminomethanphosphonsäurediethylester (Example 10) and N-thymin-1-yl-acetyl-N- (4-methoxytriphenylmethoxy) ethylaminomethaneph-osphon acid (4-nitrophenylethyl) monoester (triethylammonium salt) (Example 15). Instead of TSNT, 3-nitro-1- (2,4,6 triisopropylphenylsulfonyl) -1H-1,2,4-triazole (TIPSNT) used for the coupling. For cleaning was over silica gel chromatographed (EA / methanol / TEA 95/5/2, then 90/10/2). Yield:) 90%.

MS (ES +): 1109.0 (M + Li) ⁺. 1 H-NMR (200MHz, DMSO, TMS) characteristic Signals are: Ar-H & Thymine-H: 6.82-8.18 (m, 20H); CO-CH₂: 4.51-4.76 (m, 4H); Thymine CH₃: 1.61-1.78 (m, 6H).

22) 5'-MMTr-T-P (ONPE) -T-P (OEt) ₂

The synthesis was carried out analogously to Example 21 from N-thymin-1-yl-acetyl-N- (2-hy droxy) ethylaminomethanphosphonsäurediethylester (Example 10) and N-thymin-1-yl-acetyl-N- (4-methoxytriphenylmethoxy) ethylaminomethaneph-osphon acid (4-nitrophenylethyl) monoester (triethylammonium salt) (Example 15). Instead of TSNT, 3-nitro-1- (2,4,6-triisopropylphenylsulfonyl) -1H-1,2,4-triazole (TIPSNT) used for the coupling. Yield: <90%.

Spectroscopic data see example 21.

23) 5'-HO-T-P (ONPE) -T-P (OEt) ₂

The synthesis was carried out analogously to Example 4 from "5′-MMTr-T-P (ONPE) -T-P (OEt) ₂" (Example 22). For purification, it was chromatographed on silica gel (EE / methanol / TEA 90/10/2, then 80/20/2). The yield was 75%.

MS (ES +): 836.3 (M + Li) ⁺. 1 H-NMR (200MHz, DMSO, TMS) characteristic  Signals are: Ar-H & Thymine-H: 7.11-8.22 (m, 6H); CO-CH₂: 4.55-4.77 (m, 4H); Thymine-CH₃: 1.71 (s, broad, 6H).

24) 5'-HO-T-P (OH) -T-P (OEthyl) ₂

10 mg (0.012 mmol) "5'-HO-T-P (ONPE) -T-P (OEt) ₂" (Example 23) were in 1 ml a 0.5M solution DBU dissolved in pyridine and first 24 h at 4 ° C, then 24 h stirred at room temperature. The solvent was evaporated in vacuo, the residue digested twice with pentane, then twice with ether, then was chromatographed on silica gel for purification (EA / methanol / TEA 9/1 / 0.2, then 70/30/2, then 60/40/2). Yield: 10.2 mg.

MS (FAB): 725.3 (M + 2Na-H) ⁺; 703.3 (M + Na) ⁺. 1 H-NMR (200MHz, DMSO, TMS) Characteristic signals are: Thymine-H: 7.15-7.70 (m, 2H); CO-CH₂: 4.67-4.92 (m, 4H); Thymine CH₃: 1.67-1.81 (m, 6H).

25) 5'-MMTr-T-P (ONPE) -T-P (ONPE) -T-P (OEt) ₂

The synthesis was carried out analogously to Example 17 from "5'-HO-T-P (ONPE) -T-P (OEt) ₂" (Example 22) and N-thymin-1-yl-acetyl-N- (4-methoxytriphenylmethoxy) ethylaminomethanephosphonic acid- (4-nitrophenylethyl) monoester (Triethylammonium salt) (Example 15) with the addition of 1.5 eq (based on Example 23) 4-methoxypyridine N-oxide. For cleaning was over silica gel chromatographed (EA / methanol / TEA 90/10/2, then 85/15/2). Yield: 61%.

MS (ES +): 1555.8 (M + H) ⁺. 1 H-NMR (200MHz, DMSO, TMS) characteristic Signals are: Ar-H & Thymin-H: 6.83-8.20 (m, 25H); CO-CH₂: 4.52-4.75 (m, 6H); Thymine CH₃: 1.61-1.78 (m, 9H).  

26) 5'-HO-T-P (ONPE) -T-P (ONPE) -T-P (OEt) ₂

The synthesis was carried out analogously to Example 4 from "5′-MMTr-T-P (ONPE) -T-P (ONPE) -T- P (OEt) ₂ "(Example 25). For purification, it was chromatographed on silica gel (EE / methanol / TEA 70/30/2). The yield was 89%.

MS (ES +): 1283.1 (M + H) ⁺; 1305.0 (M + Na⁺).

27) 5'-MMTr-T-P (ONPE) -T-P (ONPE) -T-P (ONPE) -T-P (OEt) ₂

The synthesis was carried out analogously to Example 17 from "5'-HO-T-P (ONPE) -T-P (ONPE) -T- P (OEt) ₂ "(Example 26) and N-thymin-1-yl-acetyl-N- (4-methoxytriphenylmethoxy) ethylaminomethanephosphonic acid- (4-nitrophenylethyl) monoester (Triethylammonium salt) (Example 15) with the addition of 1.5 eq (based on Example 23) 4-methoxypyridine N-oxide. For cleaning was over silica gel chromatographed (EA / methanol / TEA 90/10/2, then 80/20/2). Yield: 15%.

MS (ES +): 2007 (M + H) ⁺; 2029 (M + Na) ⁺. 1 H-NMR (200MHz, DMSO, TMS) Characteristic signals are: Ar-H & Thymin-H: 6.79-8.21 (m, 30H); CO-CH₂: 4.53-4.87 (m, 8H); Thymine CH₃: 1.58-1.89 (m, 12H).

28) 5'-HO-T-P (ONPE) -T-P (ONPE) -T-P (ONPE) -T-P (OEt) ₂

The synthesis was carried out analogously to Example 4 from "5′-MMTr-T-P (ONPE) -T-P (ONPE) -T- P (ONPE) -T-P (OEt) ₂ "(Example 27). For cleaning, silica gel chromatographed (EE / methanol / TEA 70/30/2). The yield was 55%.

MS (FAB): 1735 (M + H) ⁺; 1757 (M + Na) ⁺.  

29) 5'-MMTr-T-P (ONPE) -T-P (ONPE) ₂

The synthesis was carried out analogously to Example 17 from N-thymin-1-yl-acetyl-N- (2-hy droxyethyl) aminomethanephosphonic acid di (4-nitrophenylethyl) ester (example 20) and N-thymin-1-yl-acetyl-N- (4-methoxytriphenylmethoxy) ethylamino methanephosphonic acid (4-nitrophenylethyl) monoester (triethylammonium salt) (Example 15). For purification, it was chromatographed on silica gel (EE / methanol / TEA 100/0/1, then 90/10/1). Yield: 87%.

MS (FAB): 1356.2 (M + 2Li-H) ⁺. 1 H-NMR (200MHz, DMSO, TMS) Characteristic signals are: Ar-H & Thymin-H: 6.82-8.18 (m, 28H); CO-CH₂: 4.50-4.71 (m, 4H); Thymine CH₃: 1.59-1.78 (m, 6H).

30) 5'-HO-T-P (ONPE) -T-P (ONPE) ₂

The synthesis was carried out analogously to Example 4 from "5′-MMTr-T-P (ONPE) -T-P (ONPE) ₂" (Example 29). For purification, it was chromatographed on silica gel (EE / methanol / TEA 85/15/1, then 80/20/1). The yield was 78%.

MS (ES +): 1072.7 (M + H) ⁺. 1 H-NMR (200MHz, DMSO, TMS) characteristic Signals are: Ar-H & Thymine-H: 7.08-8.20 (m, 14H); CO-CH₂: 4.52-4.80 (m, 4H); Thymine-CH₃: 1.70 (s, broad, 6H).

31) 5′-MMTr-C ^to -P (ONPE) -C ^to -P (ONPE) -C ^to -P (ONPE) ₂

The synthesis was carried out analogously to Example 17 from N- (N⁶-anisoyl) cytosin-1-yl-acetyl-N- (4-methoxytriphenylmethoxy) ethylaminomethanephosphonic acid (2- (p-nitrophenyl) ethyl) monoester (triethylammonium salt) (Example 3) and "5'-HO-C ^An - P (ONPE) -C ^An -P (ONPE) ₂" (Example 6). For purification, it was chromatographed on silica gel (EA / methanol / TEA 80/19/1). Yield: 66%.

MS (FAB): 2155 (M + H) ⁺; 2161 (M + Li) ⁺; 2177 (M + Na) ⁺.

32) 5'-HO-C ^to -P (ONPE) -C ^to -P (ONPE) -C ^to -P (ONPE) ₂

The synthesis was carried out analogously to Example IV from "5'-MMTr-C ^An -P (ONPE) -C ^An - P (ONPE) -C ^An -P (ONPE) ₂" (Example 31). For purification, it was chromatographed on silica gel (EA / methanol / TEA 85/15/1, then 80/20/1). The yield was 70%.

MS (FAB): 1882 (M + H) ⁺; 1904 (M + Na) ⁺.

33) N- (N⁶-anisoyl) cytosin-1-yl-acetyl-N- (4-methoxytriphenylmethoxy) ethyl aminomethanephosphonic acid allyl- (2- (p-nitrophenyl) ethyl) diester

The synthesis was carried out analogously to Example 17 from N- (N⁶-anisoyl) cytosin-1-yl-acetyl N- (4-methoxytriphenylmethoxy) ethylaminomethanephosphonic acid (2- (p-nitr-o phenyl) ethyl) monoester (triethylammonium salt) (Example 3) and allyl alcohol. For purification, it was chromatographed on silica gel (EA / methanol / TEA 95/5/1).

MS (ES +): 902.1 (M + H) ⁺; 924.1 (M + Na) ⁺. 1 H-NMR (200MHz, DMSO, TMS): δ = 2.94-3.70 (m, 8H, P-O-CH₂-CH₂-Ar + MMTr-O-CH₂ + N-CH₂ + P-CH₂); 3.75 (s, 3H, OCH₃); 3.86 (s, 3H, OCH₃); 4.10-4.60 (m, 4H, P-O-CH₂); 4.79 & 4.84 (each s, broad, 2H, CO-CH₂); 5.09-5.39 (m, 2H, H₂C = CH-); 5.71-6.00 (m, 1H, H₂C = CH-); 6.83-8.19 (m, 24H, Ar-H, cytosinyl-H); 11.03 (broad s, 1H, NH).  

34) N- (N⁶-anisoyl) cytosin-1-yl-acetyl-N- (2-hydroxy) ethylaminomethanephosph-hon acid allyl (2- (p-nitrophenyl) ethyl) diester

The synthesis was carried out analogously to Example 4 from N- (N⁶-anisoyl) cytosin-1-yl-acetyl N- (4-methoxytriphenylmethoxy) ethylaminomethanephosphonic acid allyl- (2 - (p-nitro phenyl) ethyl) diester (Example 33). For cleaning was over silica gel chromatographed (EA / methanol / TEA 94/5/1). The yield was 83%.

MS (ES +): 630.2 (M + H) ⁺. 1 H-NMR (200MHz, DMSO, TMS): δ = 3.02 (t, 2H, P-O-CH₂-CH₂-Ar); 3.37-3.72 (m, 4H, HO-CH₂-CH₂); 3.86 (s, 3H, OCH₃); 3.91 (d, J = 11Hz, 2H, P-CH₂); 4.22 (dt, 2H, P-O-CH₂-CH₂-Ar); 4.40 (dd, 2H, O-CH₂- CH = CH₂); 4.78 & 5.01 (m, 2H, CO-CH₂); 5.11-5.33 (m, 2H, H₂C = CH-); 5.71- 6.00 (m, 1H, H₂C = CH-); 6.99-8.21 (m, 14H, Ar-H, cytosinyl-H); 11.03 (broad s, 1H, NH).

35) N-Thymin-1-yl-acetyl-N- (4-methoxytriphenylmethoxy) ethylamino allyl methanephosphonic acid (2- (p-nitrophenyl) ethyl) diester

Synthesis analogous to Example 17 from N-thymin-1-yl-acetyl-N- (4-methoxytriphenyl methoxy) ethylaminomethanephosphonic acid- (4-nitrophenylethyl) monoester (Triethylammonium salt, Example 15) and allyl alcohol. For cleaning was over Chromatographed silica gel (EA / methanol / TEA 97/3/2). The yield was 100%.

MS (FAB): 805.3 (M + Na) ⁺.  

36) N-thymin-1-yl-acetyl-N- (2-hydroxy) ethylaminomethanephosphonic acid allyl- (2- (p-nitrophenyl) ethyl) diester

Synthesis analogous to Example 4 from N-thymin-1-yl-acetyl-N- (4-methoxy triphenylmethoxy) ethylaminomethanephosphonic acid allyl- (2- (p-nitro phenyl) ethyl) diester (Example 35). For cleaning was over silica gel chromatographed (EE / methanol / TEA 90/10/2). The yield was 86%.

MS (ES +): 511.1 (M + H) ⁺.

37) 5′-MMTr-T-P (ONPE) -T-P (ONPE) (OAllyl)

Synthesis analogous to Example 17 from N-thymin-1-yl-acetyl-N- (2-hydroxy) ethylamino methanephosphonic acid allyl- (2- (p-nitrophenyl) ethyl) diester (Example 36) and N-thymine 1-yl-acetyl-N- (4-methoxytriphenylmethoxy) ethylaminomethanephosphon acid (4-nitrophenylethyl) monoester (triethylammonium salt) (Example 15). For Purification was chromatographed on silica gel (EA / methanol / TEA 90/10/2). Yield: 90%.

MS (FAB): 1257.3 (M + Na) ⁺.

38) 5'-MMTr-T-P (ONPE) -T-P (ONPE) -T-P (ONPE) -T-P (ONPE) -T-P (OEt) ₂

Synthesis analogous to Example 17 from "5'-HO-T-P (ONPE) -T-P (ONPE) -T-P (ONPE) -T- P (OEt) ₂ "(Example 28) and N-thymin-1-yl-acetyl-N- (4-methoxy triphenylmethoxy) ethylaminomethanephosphonic acid- (4-nitrophenyl ethyl) monoester (triethylammonium salt) (Example 15). For cleaning was over Chromatographed silica gel (EA / methanol / TEA 80/20/2). Yield: 57%.

MS (FAB): 2460 (M + H) ⁺; 2482 (M + Na) ⁺.  

39) 5'-HO-T-P (ONPE) -T-P (ONPE) -T-P (ONPE) -T-P (ONPE) -T-P (OEt) ₂

Synthesis analogous to Example 4 from "5′-MMTr-T-P (ONPE) -T-P (ONPE) -T-P (ONPE) -T- P (ONPE) -T-P (OEt) ₂ "(Example 38). For cleaning, silica gel chromatographed (EE / methanol / TEA 70/30/2). The yield was 55%.

MS (FAB): 2209 (M + Na) ⁺.

40) 5'-HO-T-P (OH) -T-P (OH) -T-P (OH) -T-P (OH) -T-P (OEt) ₂

4.0 mg (0.001 83 mmol) "5′-HO-T-P (ONPE) -T-P (ONPE) -T-P (ONPE) -T-P (ONPE) -T- P (OEt) ₂ "(Example 39) were in 1.1 ml of a 0.5 molar solution of DBU in Dissolved pyridine and stirred for 24 h at room temperature. The reaction mixture was evaporated in vacuo, the residue was stirred several times with toluene. The solvent was removed with a syringe, the residue again with Pentane stirred, the solvent removed again with a syringe. The Product was dried in vacuo. The yield was 4 mg one strong hygroscopic powder.

MS (ES-): 1589.7 (MH ^- ) -; 1611.8 (M + Na-2H) ⁻.

41) 5'-MMTr-T-P (ONPE) -T-P (ONPE) -T-P (ONPE) -T-P (ONPE) -T-P (ONPE) -T-P (OEt) ₂

a) Synthesis analogous to Example 17 from "5'-HO-T-P (ONPE) -T-P (ONPE) -T-P (ONPE) -T- P (ONPE) -T-P (OEt) ₂ "(Example 39) and N-thymin-1-yl-acetyl-N- (4-methoxy triphenylmethoxy) ethylaminomethanephosphonic acid- (4-nitrophenyl ethyl) monoester (triethylammonium salt) (Example 15). For cleaning was over Chromatographed silica gel (EA / methanol / TEA 80/20/2, then 70/30/2).  

MS (FAB): 2934 (M + Na) ⁺, 2957 (M + 2Na-H) ⁺, 2978 (M + 3Na-2H) ⁺.

b) Synthesis analogous to Example 17 from "5'-HO-T-P (ONPE) -T-P (ONPE) -T-P (ONPE) -T- P (OEt) ₂ "(Example 28) and" 5'-MMTr-T-P (ONPE) -T-P (ONPE) (OH) "(Example 42). For purification, it was chromatographed on silica gel (EA / methanol / TEA 80/20/2, then 70/30/2). The target fraction was evaporated in vacuo, then triturated with pentane and ether. MS as above.

42) 5′-MMTr-T-P (ONPE) -T-P (ONPE) (OH)

24.7 mg (0.02 mmol) "5′-MMTr-T-P (ONPE) -T-P (ONPE) (OAllyl)" (Example 37) together with 16.2 mg (0.12 mmol) Diethylammonium hydrogen carbonate in 2 ml absolute. DCM solved. At 15-20 ° C a solution of 13.9 mg (0.012 mmol) of tetrakis (triphenylphosphine) palladium (0) and 2.1 mg (0.008 mmol) triphenylphosphine in 2 ml absolute. DCM added dropwise during 2 min. It was 30 min at room temp. touched. For Purification, the reaction mixture was chromatographed on silica gel (EE / methanol / TEA 80/20/1, then 60/40/1). The product fraction was in Evaporated in vacuo, the residue with pentane, then with EA / ether, then triturated again with pentane and dried in vacuo. Yield: 57%.

MS (ES-): 1193.6 (M-H) ⁻.

1 H-NMR (200MHz, DMSO, TMS): Characteristic signals: δ =; 1.67 & 1.72 (each. s, 3H, T-CH₃); 4.60 & 4.82 (each s, 2H, CO-CH₂); 6.83-8.19 (m, 24H, Ar- H, T-H).  

43) 5'-HO-T-P (ONPE) -T-P (ONPE) -T-P (ONPE) -T-P (ONPE) -T-P (ONPE) -T-P (OEt) ₂

Synthesis analogous to Example 4 from "5′-MMTr-T-P (ONPE) -T-P (ONPE) -T-P (ONPE) -T- P (ONPE) -T-P (ONPE) -T-P (OEt) ₂ "(Example 41). After the reaction, the The reaction mixture was concentrated, the residue was coevaporated three times with toluene, then first mixed with EA / ether, then with pentane. The backlog was in Vacuum dried.

MS (FAB): 2662 (M + Na) ⁺.

44) 5′-HO-T-P (ONPE) -T-P (ONPE) (OAllyl)

Synthesis analogous to example 4 from 5′-MMTr-T-P (ONPE) -T-P (ONPE) (OAllyl) (example 37). For purification, it was chromatographed on silica gel (EA / methanol / TEA 90/10/1, then 80/20/1). The yield was 87%.

MS (FAB): 963.0 (M + H) ⁺; 985.1 (M + Na) ⁺.

45) 5′-MMTr-T-P (ONPE) -T-P (ONPE) -T-P (ONPE) (OH) a) 5′-MMTr-T-P (ONPE) -T-P (ONPE) -T-P (ONPE) (OAllyl)

Synthesis analogous to Example 17 from "5'-HO-T-P (ONPE) -T-P (ONPE) (OAllyl)" (Example 44) and N-thymin-1-yl-acetyl-N- (4-methoxytriphenylmethoxy) ethylaminomethane phosphonic acid (4-nitrophenylethyl) monoester (triethylammonium salt) (example 15). For purification, it was chromatographed on silica gel (EA / methanol / TEA 90/10/1, then 85/15/1). Yield: 55%  

b) 5′-MMTr-T-P (ONPE) -T-P (ONPE) -T-P (ONPE) (OH)

"5′-MMTr-T-P (ONPE) -T-P (ONPE) -T-P (ONPE) (OAllyl)" (Example 45a) was as in Example 42 described with tetrakis (triphenylphosphine) palladium (O) reacted. For purification, it was chromatographed on silica gel (EA / methanol / TEA 80/20/2, then 70/30/2). The product fraction was evaporated in vacuo, the residue was triturated with pentane and ether. Yield: 98%.

MS (ES +; LiCl): 1654.1 (M + Li⁺).

46) 5'-MMTr-T-P (ONPE) -T-P (ONPE) -T-P (ONPE) -T-P (OEt) ₂

Synthesis analogous to Example 17 from N-thymin-1-yl-acetyl-N- (2-hydroxy) ethylamino diethyl methanephosphonate (Example 10) and "5′-MMTr-T-P (ONPE) -T- P (ONPE) -T-P (ONPE) (OH) "(Example 45b). Purification was carried out over silica gel chromatographed (EA / methanol / TEA 90/10/2, then 80/20/2). Refurbishment, Purification and characterization as in example 27.

47) 5'-MMTr-T-P (ONPE) -T-P (ONPE) -T-P (ONPE) -T-P (ONPE) -T-P (ONPE) -T-P (OEt) ₂

Synthesis analogous to Example 17 from "5′-HO-TP (ONPE) -TP (ONPE) -TP (OEt) ₂" (Example 26) and "5′-MMTr-TP (ONPE) -TP (ONPE) -TP ( ONPE) (OH) "(Example 45b). For purification, it was chromatographed on silica gel (EA / methanol / TEA 80/20/2). The product fraction was evaporated in vacuo, coevaporated with toluene and purified by preparative HPLC (High Pressure Liquid Chromatography): RP8 LiChrospher60, water / acetonitrile /: 1/1; 0.1% ammonium acetate; 1 ml / min) R _f = 12.97 min.

48) 5'-HO-T-P (OH) -T-P (OH) -T-P (OH) -T-P (OH) -T-P (OH) -T-P (OEt) ₂ a) 5'-HO-T-P (ONPE) -T-P (ONPE) -T-P (ONPE) -T-P (ONPE) -T-P (ONPE) -T-P (OEt) ₂

Synthesis analogous to Example 4 from "5′-MMTr-T-P (ONPE) -T-P (ONPE) -T-P (ONPE) -T- P (ONPE) -T-P (ONPE) -T-P (OEt) ₂ "(Example 47) Chromatographed silica gel (EA / methanol / TEA 70/30/2, then 60/40/2). The The product fraction was evaporated in vacuo, the residue first with Pentane, then stirred with ether and dried in vacuo. Yield: 100%.

MS (FAB): 2662 (M + Na) ⁺, 2684 (M + 2Na-H) ⁺, 2706 (M + 3Na-2H) ⁺.

b) 5'-HO-T-P (OH) -T-P (OH) -T-P (OH) -T-P (OH) -T-P (OH) -T-P (OEt) ₂

"5′-HO-T-P (ONPE) -T-P (ONPE) -T-P (ONPE) -T-P (ONPE) -T-P (ONPE) -T-P (OEt) ₂" (Example 48a) was implemented and worked up analogously to Example 40 with DBU.

MS (ES-): 1892 (MH ^- ) -; 1915 (M + Na-2H) ⁻.

49) 5′-MMTr-T-P (ONPE) -T-P (ONPE) -T-P (ONPE) -T-P (ONPE) -T-P (ONPE) -T- P (ONPE) -T-P (OEt) ₂

Synthesis analogous to Example 17 from "5'-HO-TP (ONPE) -TP (ONPE) -TP (ONPE) -T- P (OEt) ₂" (Example 28) and "5'-MMTr-TP (ONPE) - TP (ONPE) -TP (ONPE) (OH) "(Example 45b). For purification, it was chromatographed on silica gel (EA / methanol / TEA 80/20/2, then 70/30/2). The product fraction was evaporated in vacuo, coevaporated with toluene and purified by preparative HPLC: RP8 LiChrospher60, water / acetonitrile /: 1/1; 0.1% ammonium acetate; 1 ml / min) R _f = 15.24 min.

MS (FAB): 3386 (M + Na) ⁺, 3409 (M + 2Na-H) ⁺.

50) 5′-HO-T-P (ONPE) -T-P (ONPE) -T-P (ONPE) -T-P (ONPE) -T-P (ONPE) -T-P (ONPE) - T-P (OEt) ₂

Synthesis analogous to Example 4 from "5′-MMTr-T-P (ONPE) -T-P (ONPE) -T-P (ONPE) -T- P (ONPE) -T-P (ONPE) -T-P (ONPE) -T-P (OEt) ₂ "(Example 49). For cleaning evaporated in vacuo, coevaporated three times with toluene, the residue first with pentane, then with ether and dried in vacuo. Yield: <90%.

MS (FAB): 3114 (M + Na) ⁺.

51) 5'-HO-T-P (OH) -T-P (OH) -T-P (OH) -T-P (OH) -T-P (OH) -T-P (OH) -T-P (OEt) ₂

"5′-HO-T-P (ONPE) -T-P (ONPE) -T-P (ONPE) -T-P (ONPE) -T-P (ONPE) -T-P (ONPE) -T- P (OEt) ₂ "(Example 50) was implemented analogously to Example 40 with DBU and worked up.

MS (ES-): 2196 (M-H⁻) ⁻; 2218 (M + Na-2H) ⁻.

52) 5′-MMTr-T-P (ONPE) -T-P (ONPE) -T-P (ONPE) -T-P (ONPE) -T-P (ONPE) -T- P (ONPE) -T-P (ONPE) -T-P (ONPE) -T-P (OEt) ₂

Synthesis analogous to Example 17 from "5′-HO-TP (ONPE) -TP (ONPE) -TP (ONPE) -T- P (ONPE) -TP (ONPE) -TP (OEt) ₂" (Example 48a) and "5'-MMTr-TP (ONPE) -T- P (ONPE) -TP (ONPE) (OH) "(Example 45b). For purification, it was chromatographed on silica gel (EA / methanol / TEA 70/30/2, then 60/40/2). The product fraction was evaporated in vacuo, coevaporated with toluene and purified by preparative HPLC: RP8 LiChrospher60, water / acetonitrile /: 1/1; 0.1% ammonium acetate; 1 ml / min) R _f = 23.95 min.

53) 5′-HO-T-P (OH) -T-P (OH) -T-P (OH) -T-P (OH) -T-P (OH) -T-P (OH) -T-P (OH) -T- P (OH) -T-P (OEt) ₂ a) 5′-HO-T-P (ONPE) -T-P (ONPE) -T-P (ONPE) -T-P (ONPE) -T-P (ONPE) -T-P (ONPE) -T-- P (ONPE) -T-P (ONPE) -T-P (OEt) ₂

Synthesis analogous to Example 4 from "5′-MMTr-T-P (ONPE) -T-P (ONPE) -T-P (ONPE) -T- P (ONPE) -T-P (ONPE) -T-P (ONPE) -T-P (ONPE) -T-P (ONPE) -T-P (OEt) ₂ "(Example 52). For cleaning, it was evaporated in vacuo, coevaporated three times with toluene, the residue was stirred first with pentane, then with ether and in vacuo dried. Yield: <90%.

b) 5′-HO-T-P (OH) -T-P (OH) -T-P (OH) -T-P (OH) -T-P (OH) -T-P (OH) -T-P (OH) -T-P (OH) - T-P (OEt) ₂

"5′-HO-T-P (ONPE) -T-P (ONPE) -T-P (ONPE) -T-P (ONPE) -T-P (ONPE) -T-P (ONPE) -T- P (ONPE) -T-P (ONPE) -T-P (OEt) ₂ "(Example 53a) was analogous to Example 40 with DBU implemented and revised.

MS (ES-): 2802 (M-H⁻) ⁻; 2825 (M + Na-2H) ⁻.  

54) 2- (N'-tert-butyloxycarbonylamino) ethylaminomethanephosphonic acid di (2 - (p-nitro phenyl) ethyl) ester a) 1-methylimino-2- (N'-tert-butyloxycarbonylamino) ethane (trimer)

2.0 g (12.5 mmol) 2-amino-1- (N'-tert-butyloxycarbonylamino) ethane, dissolved in 8 ml of methanol were cooled with ice with 1.52 ml (18.72 mmol) 37% Formaldehyde added and 1 h at room temp. touched. The backlog was in EE added, twice with saturated NaHCO3 solution, then with Washed NaCl solution, dried, filtered and evaporated in vacuo. For Purification chromatographed on silica gel (EE / TEA 100 / 0.2, then EE / Methano / TEA 90/10 / 0.2). The yield was 0.8 g.

MS (FAB / LiCl) 523.4 (M + 2Li-H) ⁺. 1 H-NMR (200MHz, DMSO, TMS): δ = 1.38 (s, 27H, tBu-H); 2.40 (t, 6H, N-CH₂); 2.99 (t, 6H, N-CH₂); 3.25 (t, 6H, N-CH₂); 6.81 (t, broad, 3H, NH).

b) 2- (N'-tert-butyloxycarbonylamino) ethylaminomethanephosphonic acid di (2 - (p-nitro phenyl) ethyl) ester

1-methylimino-2- (N'-tert-butyloxycarbonylamino) ethane (trimer) (Example 54a) was analogous to Example 1f with di (2- (4-nitrophenyl) ethyl) phosphite (Example 1e) implemented. For purification, it was chromatographed on silica gel (first Toluene / EE / TEA 20/80 / 0.2; then EE / TEA 100 / 0.2, then EE / methanol / TEA 0.5 / 5 / 0.2). Yield: 25%.

MS (ES + / LiCl) 553.3 (M + H) ⁺, 559.3 (M + Li) ⁺. 1 H-NMR (200MHz, DMSO, TMS): δ = 1.37 (s, 9H, tBu-H); 2.83 (d, J = 12Hz, 2H, P-CH₂); 2.55 (t, 4H, Ar- CH₂); 2.90-3.09 (m, 4H, N-CH₂-CH₂-N); 4.16 (dt, 4H, PO-CH₂); 7.52 & 8.15 (each. d, 8H, Ar-H).  

55) N-Thymin-1-yl-acetyl-N- (2-N'-tert-butyloxycarbonylamino) ethylamino methanephosphonic acid di (2- (p-nitrophenyl) ethyl) ester

Synthesis analogous to Example 8 from 2- (N'-tert-butyloxycarbonylamino) ethylamino methanephosphonic acid di (2- (p-nitrophenyl) ethyl) ester (Example 54b) and tymidine 1-yl acetic acid. Yield: 86%

MS (FAB / LiCl): 725.3 (M + Li) ⁺. 1 H-NMR (200MHz, DMSO, TMS): δ = 1.42 (s, 9H, tBu-H); 1.91 (s, 3H, T-CH₃); 2.99-3.58 (m, 8H, P-O-CH₂-CH₂-Ar & N-CH₂- CH₂-N); 3.75 (d, J = 12Hz, 2H, P-CH₂); 4.06-4.38 (m, 4H, PO-CH₂); 7.37 & 8.15 (each d, 8H, Ar-H).

56) Interaction with DNA: UV melting curve

The interaction of the compounds according to the invention with complementary nucleic acids was exemplified by recording the UV melting curve of "5'-HO-TP (OH) -TP (OH) -TP (OH) -TP (OH) -TP (OH) -TP (OH) -TP (OH) -T- P (OH) -TP (OEt) ₂ "(Example 53b) with (dA) ₉ demonstrated. A 0.3 OD of "5'-HO-TP (OH) -TP (OH) -TP (OH) -TP (OH) -TP (OH) -TP (OH) -T- P (OH) - TP (OH) -TP (OEt) ₂ "and (dA) ₉ in 1 ml of a buffer (1 M NaCl, 20 mM MgCl₂, 10 mM HEPES, pH 7.5) and the change in absorbance at 260 nm depending on the Temperature (0 to recorded. The result can be seen in Fig. 1. A T _m value of approximately 23 ° C. was determined from the melting curve obtained.

57) Interaction with DNA: gel shift experiment

The interaction of the compounds of the invention with complementary  Nucleic acids were exemplified by the hybridization of "5'-HO-T-P (OH) - T-P (OH) -T-P (OH) -T-P (OH) -T-P (QH) -T-P (OH) -T-P (OH) -T-P (OH) -T-P (OEt) ₂ " (Example 53b) demonstrated with (dA) demonstr in a gel shift experiment. To do this "5′-HO- T-P (OH) -T-P (OH) -T-P (OH) -T-P (OH) -T-P (OH) -T-P (OH) -T-P (OH) -T-P (OH) -T- P (OEt) ₂ "(Example 53b) and (dA) ₉ each alone and mixed in a ratio of 1: 1, 1: 2, 1: 5 and 1:10 on a non-denaturing polyacrylamide gel (20%, Running buffer 1 × TBE, 10 mM MgCl₂) applied and the running behavior determined. The result can be seen in Fig. 2: (dA) ₉ runs faster than "5'-HO-T-P (OH) -T- P (OH) -T-P (OH) -T-P (OH) -T-P (OH) -T-P (OH) -T-P (OH) -T-P (OH) -T-P (OEt) ₂ ", in which 1: 1 mix are both only weakly visible, but this is a slower one Gang emerged that was a complex between the two components corresponds. In the 2: 1 mixture of (dA) ₉ nothing is to be seen anymore, that's why new bonds all the more clearly. The same applies to the 5: 1 or 10: 1 mixture, in which the significant excess of "5′-HO-T-P (OH) -T-P (OH) -T- P (OH) -T-P (OH) -T-P (OH) -T-P (OH) -T-P (OH) -T-P (OH) -T-P (OEt) ₂ "can be seen.

58) 5'-MMTr-T-P (ONPE) -T-P (OEthyl) ₂ [s. also example 21, alternative Syntheses]

a) 8.44 mg (10 µmol) N-thymin-1-yl-acetyl-N- (4-methoxy triphenylmethoxy) ethylaminomethanephosphonic acid- (4-nitrophenylethyl) monoester (triethylammonium salt) (Example 15), 3.77 mg (10 µmol) N -Thymin-1-yl-acetyl-N- (2-hydroxy) ethylaminomethanephosphonic acid diethyl ester (Example 10) and 64.6 mg (500 µmol) N-ethyldiisopropylamine (DIPEA) were in 0.3 ml absolute. DMF solved. 44.2 mg (100 μmol) of benzotriazol-yloxy) tris (dimethylamino) phosphonium hexafluorophosphate (BOP) were added. It was 24 h at room temp. touched. After TLC (EA / methanol / TEA 100/20/2; R _f = 0.5) approx. 70% yield.

b) Analogous to example 58a, but with 30 μmol HATU (O- (7-azabenzotriazol-1yl) - N, N, N ′, N′-tertamethyluronium hexafluorophosphate) instead of 100 µmol BOP. After TLC approx. 65% yield.

59) N-Thymin-1-yl-acetyl-N- (4-methoxytriphenylmethoxy) ethylamino methanephosphonic acid [2- (p-nitrophenyl) ethyl] - [5 ′ - (3′-levuloyl-thymid-in] diester

Synthesis analogous to Example 17 from N-thymin-1-yl-acetyl-N- (4-methoxytriphenyl methoxy) ethylaminomethanephosphonic acid- (4-nitrophenylethyl) monoester (Triethylammonium salt, Example 15) and 3'-levuloyl-thymidine. For cleaning was chromatographed on silica gel (DCM / methanol / TEA 98/2 / 0.25). The Yield was 46%.

MS (FAB / LiCl): 1071.4 (M + Li) ⁺.

60) N-Thymin-1-yl-acetyl-N- (4-methoxytriphenylmethoxy) ethylamino methanephosphonic acid [2- (p-nitrophenyl) ethyl] - [5'-thymidine] diester

64 mg (0.06 mmol) of N-thymin-1-yl-acetyl-N- (4-methoxy triphenylmethoxy) ethylaminomethanephosphonic acid- [2- (p-nitro phenyl) ethyl] - [5 ′ - (3′-levuloyl-thymidine] diesters (Example 59) were added in 0.5 ml Dioxane dissolved. 9 mg (0.23 mmol) NaBH₄ in 0.12 ml water were added and stirring was carried out at room temperature for 20 minutes. The solvent was in Evaporated in vacuo, the residue taken up in DCM, with water extracted and dried. For purification, it was chromatographed on silica gel (DCM / methanol / TEA 92/8 / 0.5). The yield was 72%.

MS (FAB / LiCl): 973.4 (M + Li) ⁺, 979.4 (M + 2Li-H) ⁺, 985.4 (M + 3Li-2H) ⁺.  

61) N-Thymin-1-yl-acetyl-N- (4-methoxytriphenylmethoxy) ethylamino methanephosphonic acid- [2- (p-nitrophenyl) ethyl] - [5′-thymidine- (3 ′ - (cyano-ethyl-N, N-di isopropyl phosphoramidite)] diester

31 mg (0.032 mmol) of N-thymin-1-yl-acetyl-N- (4-methoxytriphenylmethoxy) ethylaminomethanephosphonic acid- [2- (p-nitrophenyl) ethyl] - [5'-thymidine] diesters (Example 60) were twice with absolute. CH₃CN coevaporated, in 0.4 ml absolute. THF solved. 12.4 mg (0.096 mmol) diisopropylethylamine, then 9.8 mg (0.045 mmol) of cyanoethyl chloro-diisopropyl phosphoramidite admitted. The mixture was stirred for 3 h, filtered off and evaporated in vacuo. The Yield was 63%.

MS (FAB / LiCl): 1173.3 (M + Li) ⁺, 1180.4 (M + 2Li-H) ⁺, 1186.4 (M + 3Li-2H) ⁺.

62) N- [N9- (O6-diphenylcarbamoyl-N2-acetylguanine] acetyl-N- (4-methoxy triphenylmethoxy) ethylaminomethanephosphonic acid di (2- (p-nitrophenyl) et-hyl) ester

Synthesis analogous to Example 2 from N- (4-methoxytriphenylmethoxy) ethylamino methanephosphonic acid di (2- (p-nitrophenyl) ethyl) ester (Example 1f) and O6-diphenylcarbamoyl-N2-acetylguanine acetic acid. For cleaning was over Chromatographed silica gel (EE / TEA 98/2). Yield: 87%

MS (FAB / LiCl): 1154.8 (M + H) ⁺, 1160.7 (M + Li) ⁺.

63) N- [N9- (N4-Anisoyladenin] acetyl-N- (4-methoxytriphenylmethoxy) ethylamino methanephosphonic acid di (2- (p-nitrophenyl) ethyl) ester

Synthesis analogous to Example 2 from N- (4-methoxytriphenylmethoxy) ethylamino  methanephosphonic acid di (2- (p-nitrophenyl) ethyl) ester (Example 1f) and N4-Anisoyladeninetic acid. For purification, it was chromatographed on silica gel (DCM / methanol / TEA 95/4/1). Yield: 82%.

MS (ES +): 1035.7 (M + H) ⁺.

1 H-NMR (200 MHz, DMSO, TMS): δ = 2.93 (t, 4H, P-O-CH₂-CH₂-Ar); 3.09 (t, 2H, MMTr-O-CH₂); 3.23-3.75 (m, 4H, P-CH₂ + N-CH₂); 3.75 (s, 3H, OCH₃); 3.87 (s, 3H, OCH₃); 4.08 (dt, 4H, P-O-CH₂); 5.28-5.42 (m, 2H, CO-CH₂); 6.81-8.20 (m, 28H, Ar-H, A-H); 11.00 (broad s, 1H, NH).

64) N- [N9- (N4-anisoyladenin] acetyl-N- (4-methoxytriphenylmethoxy) ethylamino-no methanephosphonic acid mono (2- (p-nitrophenyl) ethyl) ester

Synthesis analogous to Example 3 from N- [N9- (N4-anisoyladenin] acetyl-N- (4-methoxy triphenylmethoxy) ethylaminomethanephosphonic acid di (2- (p-nitrophenyl) et-hyl) ester (Example 63). For purification, it was chromatographed on silica gel (EE / methanol / TEA 65/35/2). The yield was 52%.

MS (FAB / LiCl): 874.3 (M + 2Li-H) ⁺.

65) N-Thymin-1-yl-acetyl-N- (2-methoxy) ethylaminomethanephosphonic acid di (2 - (p-nitro phenyl) ethyl) ester

Synthesis analogous to Example 2 from N- (2-methoxy) ethylaminomethanephosphon acid di (2- (p-nitrophenyl) ethyl) ester (prepared analogously to Example 1 starting from 2-methoxyethylamine by reaction with formaldehyde and di (2- (4-nitro phenyl) ethyl) phosphite) and tymidin-1-yl acetic acid. Was for cleaning Chromatographed on silica gel (EA / methanol 90/10). Yield: 64%

MS (FAB / LiCl): 640.3 (M + Li) ⁺.  

66) 5′-MMTr-C ^An -P (ONPE) -C ^A nP (ONPE) (OAllyl)

Synthesis analogous to Example 17 from N- (N⁶-anisoyl) cytosin-1-yl-acetyl-N- (4-methoxy triphenylmethoxy) ethylaminomethanephosphonic acid (2- (p-nitro phenyl) ethyl) monoester (triethylammonium salt) (Example 3) and N- (N⁶-anisoyl) cytosin-1-yl-acetyl-N- (2-hydroxy) ethylaminomethanephosphonic acid allyl- (2- (p-nitrophenyl) ethyl) diester (Example 34). For cleaning was over silica gel chromatographed (EA / methanol / TEA 85/14/1). Yield: 36%.

MS (FAB): 1474 (M + H) ⁺; 1496 (M + Na) ⁺.

67) 5′-HO-C ^An -P (ONP _{E) -C} ^An -P (ONPE) _(OAllyl)

Synthesis analogous to Example 4 from "5'-MMTr-C ^An -P (ONPE) -C ^A nP (ONPE) (OAllyl)" (Example 66). For purification, it was chromatographed on silica gel (EA / methanol / TEA 80/19/1). The yield was 55%.

MS (FAB): 1201.3 (M + H) ⁺, 1223.3 (M + Na) ⁺.

68) 5′-MMTr-C ^An -P (ONPE) -C ^A nP (ONPE) -C ^A nP (ONPE) (OAllyl)

Synthesis analogous to Example 17 from N- (N⁶-anisoyl) cytosin-1-yl-acetyl-N- (4-methoxy triphenylmethoxy) ethylaminomethanephosphonic acid (2- (p-nitrophenyl) ethyl) monoester (triethylammonium salt) (Example 3) and 5 '-HO-C ^An - P (ONPE) -C ^An -P (ONPE) (OAllyl) (Example 67). For purification, it was chromatographed on silica gel (EA / methanol / TEA 80/20/1). Yield: 58%.

MS (FAB): 2046 (M + H) ⁺; 2068 (M + Na) ⁺.  

69) 5′-MMTr-C ^An -P (ONPE) -C ^A nP (ONPE) -C ^A nP (ONPE) (OH)

Synthesis analogous to Example 42 from 5'-MMTr-C ^An -P (ONPE) -C ^A nP (ONPE) -C ^A n- P (ONPE) (OAllyl) (Example 68). For purification, it was chromatographed on silica gel (EA / methanol / TEA 60/38/2). Yield: 66%.

MS (FAB): 2027 (M + Na) ⁺; 2049 (M + 2Na-H) ⁺.

70) 5′-MMTr-C ^An -P (ONPE) -C ^A nP (ONPE) -C ^A nP (ONPE) -C ^A nP (ONPE) -C ^A n- P (ONPE) -C ^An -P ( ONPE) ₂

Synthesis analogous to Example 17 from "5'-MMTr-C ^An -P (ONPE) -C ^A nP (ONPE) -C ^A n- P (ONPE) (OH)" (Example 69) and "5'-HO-C ^An -P (ONPE) -C ^A nP (ONPE) -C ^A n- P (ONPE) ₂ "(Example 32). For purification, it was chromatographed on silica gel (EA / methanol / TEA 70/30/2). Yield: 58%.

MS (FAB): 3892 (M + Na) ⁺; 3914 (M + 2Na-H) ⁺.

71) 5′-MMTr-TP (ONPE) -TP (ONPE) -TP (ONPE) -C ^An -P (ONPE) -C ^A nP (ONPE) -C ^A n- P (ONPE) ₂

Synthesis analogous to Example 17 from "5'-MMTr-TP (ONPE) -TP (ONPE) -T- P (ONPE) (OH)" (Example 45) and "5'-HO-C ^A nP (ONPE) -C ^A nP (ONPE) -C ^A n- P (ONPE) ₂ "(Example 32). For purification, it was chromatographed on silica gel (EA / methanol / TEA 60/40/2). The product fraction was evaporated in vacuo and purified by preparative HPLC (High Pressure Liquid Chromatography): RP8 LiChrospher60, water / acetonitrile /: 1/1; 0.1% ammonium acetate; 1 ml / min) R _f = 16.6 min.

MS (FAB): 3534 (M + Na) ⁺; 3556 (M + 2Na-H) ⁺.

72) 5'-MMTr-TP (OH) -TP (OH) -TP (OH) -C ^An -P (OH) -C ^A nP (OH) -C ^A nP (OH) ₂

Synthesis analogous to example 40 from "5′-MMTr-T-P (ONPE) -T-P (ONPE) -T-P (ONPE) - CAN-P (ONPE) -CAn-P (ONPE) -CAn-P (ONPE) ₂ "(2 mg) (Example 71).

MS (ES-): 2466.4 (M-H).

73) 5'-MMTr-T-P (OH) -T-P (OH) -T-P (OH) -C-P (OH) -C-P (OH) -C-P (OH) ₂

Approx. 1 mg "5'-MMTr-T-P (OH) -T-P (OH) -T-P (OH) -CAn-P (OH) -CAn-P (OH) -CAn- P (OH) ₂ "(Example 72) were mixed with 3 ml of 33% aqueous NH₄OH and 24 h stirred at room temperature. The reaction mixture was in vacuo evaporated. Yield: approx. 0.6 mg (19 OD).

MS (ES-): 2064.5 (M-H) ⁻.

74) 5'-HO-T-P (OH) -T-P (OH) -T-P (OH) -C-P (OH) -C-P (OH) -C-P (OH) ₂

8 OD of "5'-MMTr-TP (OH) -TP (OH) -TP (OH) -CP (OH) -CP (OH) -CP (OH) ₂" (Example 73) were dissolved in 0.5 ml of water and applied to a PolyPak ^™ (Glen Research, # 60-1100-10). The MMTr group was spun off based on the information provided by the manufacturer (Glen Research User Guide). Yield: approx. 0.35 mg (11 OD).

MS (ES-): 1792.6 ((M-H) ⁻.  

75) 5′-MMTr-C ^An -P (ONPE) -C ^A nP (ONPE) -C ^A nP (ONPE) -TP (ONPE) -TP (ONPE) - TP (ONPE) ₂

Synthesis analogous to Example 17 from "5'-MMTr-C ^An -P (ONP _{E) -C} ^An -P (ONP _{E) -C} ^An - P (ONPE) (OH)" (Example 69) and "5'-HO -TP (ONPE) -TP (ONPE) -TP (OEt) ₂ "(Example 26). For purification, it was chromatographed on silica gel (EA / methanol / TEA 80/20/2, then 70/30/2). The product fraction was coevaporated with toluene, evaporated in vacuo and triturated with pentane. Yield: 48%.

MS (FAB): 3744 (M + Na) ⁺; 3766 (M + 2Na-H) ⁺.

Claims (9)

1. Compounds of formula I.
wherein n represents a number from zero to 100;
B independently of one another hydrogen, hydroxy, (C₁-C₂₀) alkyl, (C₁-C₂₀) alkoxy, (C₁-C₂₀) alkylthio, (C₆-C₂₀) aryl, (C₆-C₂₀) aryl- (C₁- C₆) alkyl, (C₆- C₂₀) aryl- (C₁-C₆) alkoxy, (C₆-C₂₀) aryl- (C₁-C₆) alkylthio, means an aromatic group or a heterocyclic group, where alkyl, aryl and / or the aromatic or heterocyclic group optionally one or more times by hydroxy, (C₁-C₄) alkoxy, -NR⁹R¹⁰, -C (O) OH, oxo, -C (O) OR⁸, -C (O) NR⁹R¹⁰, - CN, -F, -Cl, -Br, -NO₂, (C₂-C₆) alkoxyalkyl, -S (O) _m R⁸, - (C₁-C₆) alkyl-S (O) _m R⁸, -NHC (= NH) NHR⁸, -C (= NH) NHR⁸, -NR⁹C (= O) R⁸, = NOR⁸, NR⁹C (= O) OR¹⁰, -OC (= O) NR⁹R¹⁰ and
NR⁹C (= O) NR⁹R¹⁰ may be substituted, or
B represents a natural nucleobase, an unnatural nucleobase or a reporter ligand;
AB can also stand for a D- or L-amino acid condensed on the carboxyl group or for peptides consisting of these amino acids with a length of up to 5 amino acid residues,
L independently of one another N or R¹N⁺, and
R¹ is hydrogen or (C₁-C₆) alkyl, which may be substituted by hydroxy, (C₁-C₆) alkoxy, (C₁-C₆) alkylthio or amino, preferably hydrogen or methyl;
A independently of one another denotes a single bond, a methylene group or a group of the formula IIa or IIb; Y ′ represents = O, = S, = CH₂, = C (CH₃) ₂ or = N⁺R¹, where R¹ is as defined above;
M represents a single bond, -O-, -S- or -N⁺R¹-, where R¹ is as defined above;
R² and R³ independently of one another are hydrogen, hydroxy, (C₁-C₈) alkoxy, (C₁-C₆) alkylthio, amino, halogen, such as F, Cl, Br or (C₁-C₆) alkyl, which optionally with hydroxy , (C₁-C₆) alkoxy or (C₁-C₆) alkylthio may be substituted;
p and q are independently zero to 5;
r and s are independently zero to 5;
D and G represent CR⁵R⁶;
R⁵ and R⁶ independently of one another hydrogen, (C₁-C₆) alkyl, (C₆-C₂₀) aryl, (C₆- C₂₀) aryl- (C₁-C₆) alkyl, hydroxy, (C₁-C₆) alkoxy, ( C₁-C₆) alkylthio, and alkyl and aryl may optionally be substituted with SR¹ or NR¹R¹, where R¹ is as defined above and R¹, independently of R¹, has the same meaning as R¹;
X represents -O-, -S- or -NR¹-, wherein R¹ is as defined above;
Y represents = O or = S;
Z represents -OR⁸, -NR⁹R¹⁰ or X'Q '', where X 'are defined as X and Q''asQ;
R⁸ is hydrogen, (C₁-C₁₈) alkyl, (C₂-C₁₈) alkenyl, (C₃-C₁₈) alkynyl, (C₆-C₁₂) aryl, (C₆-C₁₂) aryl- (C₁-C₆) alkyl means, where alkyl can be substituted one or more times with hydroxy, (C₁-C₄) alkoxy, F, Cl, Br and aryl 1-3 times with hydroxy, (C₁-C₄) alkoxy, (C₁-C₄) alkyl, F, Cl, Br, NO₂, -NR⁹R¹⁰, -C (O) OH, -C (O) O- (C₁-C₆) alkyl, -C (O) NR⁹R¹⁰, may be substituted, but preferably for hydrogen, ( C₁-C₆) alkyl, (C₆-C₁₂) aryl or (C₆- C₁₂) aryl- (C₁-C₆) alkyl, where aryl is simply with (C₁-C₄) alkoxy, (C₁- C₄) - Alkyl, F, Cl, Br, NO₂, may be substituted, particularly preferably hydrogen, (C₁-C₆) alkyl, phenyl or 2- (4-nitrophenyl) ethyl;
R⁹ and R¹⁰ are independently hydrogen, (C₁-C₁₈) alkyl, (C₁-C₁₈) alkenyl, (C₁-C₁₈) alkynyl, (C₆-C₁₂) aryl, (C₆-C₁₂) aryl- (C₁ -C₆) alkyl, where alkyl can be substituted one or more times with hydroxy, (C₁-C₄) alkoxy, F, Cl, Br, or R⁹ and R¹⁰ together with the N atom carrying them can be a 4-7-membered ring form;
Q and Q 'independently of one another are hydrogen or R⁸, stand for conjugates which have a favorable influence on the properties of antisense oligonucleotides or of triple helix-forming oligonucleotides or serve as a label for a DNA probe or in the hybridization of the oligonucleotide analog to the target nucleic acid with binding or cross-linking, or oligonucleotides that may be unmodified or modified. 2. Compounds of formula I according to claim 1, wherein
n represents a number from zero to 50;
B independently represents a natural nucleobase or an unnatural nucleobase;
LN means;
A represents a group of formula IIb, wherein
r = 1 and s zero, and R², R³ = H and Y ′ = O and M represent a single bond;
D and G mean CHR⁵;
R⁵ represents hydrogen;
X represents -O-;
Y = O;
Z represents hydroxy, methoxy, ethoxy, (4-nitrophenol) ethoxy, propoxy, isoPropoxy, butoxy, pentoxy, phenoxy or allyloxy;
Q and Q 'independently of one another represent hydrogen, R⁸ or for oligonucleotides, which can be unmodified and modified, where

• a) the 3'- and / or 5'-phosphoric acid diester bridges completely or partially by phosphorothioate, phosphorus dithioate, NR⁴R⁴′-phosphoramidate, phosphate-O-methyl ester, phosphate-O-ethyl ester, phosphate-O-isopropyl ester, Methylphosphonate or phenylphosphonate bridges are replaced;
• b) one, two or three 3'- or 5-phosphoric diester bridges at the pyrimidine positions and at the 5'-end and / or at the 3'-end are replaced by formacetals and / or 3'-thioformacetals;
• c) the sugar phosphate backbone is completely or partially replaced by "PNAs" or PNA-DNA hybrids;
• d) the β-D-2'-deoxyribose units completely or partially by 2'-F-2'-deoxyribose, 2'-O- (C₁-C₆) alkyl ribose, 2'-O- (C₂-C₆) alkenyl -Ribose, 2'-NH₂-2'-deoxy ribose are replaced;
• e) the natural nucleoside bases completely or partially by 5- (C₁-C₆) alkyl uracil, 5- (C₂-C₆) alkenyl uracil, 5- (C₂-C₆) alkynyl uracil, 5- ( C₁-C₆) alkyl cytosine, 5- (C₂-C₆) alkenyl cytosine, 5- (C₂-C₆) alkynyl cytosine, 5-fluorouracil, 5-fluorocytosine, 5-chlorouracil, 5-chlorocytosine, 5 Bromuracil, 5-bromocytosine, 7-deaza-7- (C₂-C₇) alkynyl guanine, 7-deaza-7- (C₂-C₇) alkynyladenine, 7-deaza-7- (C₂-C₇) alkenyl guanine, 7-Deaza-7- (C₂-C₇) alkenyladenine, 7-Deaza-7- (C₁-C₇) alkyl guanine, 7-Deaza-7- (C₁-C₇) alkyladenine, 7-Deaza-7-bromoguanine , 7-Deaza-7-bromadenin are replaced.

3. Compounds of formula I according to claims 1 or 2, characterized in that
n represents a number from 0 to 30;
Q and Q ′ independently of one another are hydrogen, R⁸, where R⁸ is H, (C₁-C₆) alkyl, phenyl or 2- (4-nitrophenylethyl), or are oligonucleotides which can be unmodified and modified, where

• a) the 3'- and / or 5'-phosphoric diester bridges are completely or partially replaced by phosphorothioate, phosphorodithioate or methylphosphonate bridges;
• b) one, two or three 3'- or 5-phosphoric diester bridges at the 5'- and at the 3'-end are replaced;
• c) the sugar phosphate backbone is completely or partially replaced by "PNAs" or PNA-DNA hybrids;
• d) the β-D-2'-deoxyribose units completely or partially by 2'-F-2'-deoxyribose, 2'-O- (C₁-C₄) alkyl ribose, 2'-O- (C₂-C₄) alkenyl -Ribose, 2'-NH₂-2'-deoxy ribose are replaced;
• e) the natural nucleoside bases completely or partially by 5- (C₃-C₆) alkyl uracil, 5- (C₂-C₆) alkenyl uracil, 5- (C₂-C₆) alkynyl uracil, 5- ( C₁-C₆) alkyl cytosine, 5- (C₂-C₆) alkenyl cytosine, 5- (C₂-C₆) alkynyl cytosine, 7-deaza-7- (C₂-C₇) alkynyl guanine, 7- Deaza-7- (C₂-C₇) alkynyladenine, 7-Deaza-7- (C₂-C₇) alkenylguanine, 7-Deaza-7- (C₂-C₇) alkenyladenine, 7-Deaza-7- (C₁-C₁ ) -alkylguanine, 7-deaza-7- (C₁-C₇) -alkyl adenine, 7-deaza-7-bromoguanine, 7-deaza-7-bromadenine are replaced.

4. Compounds of formula I according to claims 1 to 3, characterized in that
n represents a number from 0 to 25;
B independently represents a natural nucleobase;
Z represents hydroxy, ethoxy, (4-nitrophenol) ethoxy or phenoxy;
Q and Q ′ independently of one another are hydrogen, R⁸, where R⁸ is H, (C₁-C₆) alkyl, phenyl or 2- (4-nitrophenylethyl), or are oligonucleotides which can be unmodified and modified, where

• a) the 3'- and / or 5'-phosphoric diester bridges are completely or partially replaced by phosphorothioate bridges;
• c) the sugar phosphate backbone is completely or partially replaced by "PNAs" or PNA-DNA hybrids;
• d) the β-D-2'-deoxyribose units are completely or partially replaced by 2'-O-methyl-, 2'-O-allyl-, 2'-O-butylribose;
• e) the natural nucleoside bases completely or partially by 5-hexynylcytosine, 5-hexynyluracil, 5-hexynylcytosine, -deaza-7-propynylguanine, 7-deaza-7-propynyladenine, 7-deaza-7-methylguanine, 7-deaza- 7-methyladenine, 7-deaza-7-propynyladenine, 7-deaza-7-bromoguanine, 7-deaza-7-bromadenine are replaced.

5. A process for the preparation of compounds of formula I, characterized in that
a₁) compounds of formula III wherein
D, G, L and X have the meanings given above and
S 1 represents a suitable protective group, such as, for example, dimethoxytrityl, monomethoxytrityl, trityl, pixyl, tert-butyloxycarbonyl or fluorenymethyloxycarbonyl,
with compounds of formula IV wherein
R⁵ and R⁶ have the meanings given above,
in a suitable organic solvent, at temperatures from 0 ° C. to 100 ° C., converts to compounds of the formula Va or Vb b₁) Compounds of the formula Va or Vb with compounds of the formula VIa or VIb wherein
Y is as defined above,
X ′ and X ′ ′ are independently defined as X,
S² and S³ independently of one another represent protective groups, and
L¹ represents a leaving group, preferably (C₁-C₄) alkyl,
in a suitable organic solvent, at temperatures from 0 ° C. to 100 ° C., optionally with the addition of bases, complex bases or uncharged, peralkylated polyamino-phosphazene bases, to give compounds of the formula VII wherein
D, G, L, R⁵, R⁶, S¹, S², S³, X, X ′, X ′ ′ and Y are as defined above;
c₁) compounds of the formula VII with compounds of the formula VIII, wherein
A has the meaning given above,
B ^{PR has} the same meaning as B, but may be in a protected form, and
L² stands for a leaving group known to the person skilled in the art or, if A has the meaning of formula IIb, can also stand for OH;
in a suitable organic solvent, at temperatures from -20 ° C to 100 ° C, optionally with the addition of bases, complex bases or uncharged, peralkylated polyamino-phosphazene bases or without base addition and with the addition of a coupling reagent customary for the formation of peptide bonds , converts to compounds of the formula IX wherein
A, B ^PR , D, G, L, R⁵, R⁶, S¹, S², S³, X, X ′, X ′ ′ and Y are as defined above;
d₁) splitting off the protective group S³ from compounds of the formula IX by known processes, giving compounds of the formula X. wherein
A, B ^PR , D, G, L, R⁵, R⁶, S¹, S², X ′, X ′ ′ and Y are as defined above;
e₁) from compounds of formula IX, the protecting group S¹ according to known methods, whereby compounds of formula XI are obtained wherein
A, B ^PR , D, G, L, R⁵, R⁶, S², S³, X, X ′, X ′ ′ and Y are as defined above;
f₁) Compounds of formula XI with compounds of formula X according to the "phosphotriester method" known from oligonucleotide chemistry in a suitable organic solvent, at temperatures from -20 ° C to 100 ° C, with the addition of a coupling reagent or a compound of Formula XII wherein
R¹⁵ for (C₆-C₁₂) aryl, optionally substituted one to four times by (C₁-C₆) alkyl, (C₁-C₆) alkoxy, nitro, chlorine, bromine and where one to 3 C atoms are optionally substituted by heteroatoms , and
R¹⁶ represents a leaving group,
optionally with the addition of a catalyst, it being possible for the coupling reagents to be prepared in situ, or else separately, and for the solution of the activated species to be added in a suitable solvent,
converts to compounds of formula XIII, wherein
A, B ^PR , D, G, L, R⁵, R⁶, S¹, S², S³, X, X ′, X ′ ′ and Y are as defined above;
g₁) starting from compounds of the formula XIII, the steps e₁) and f₁) are repeated to the desired chain length, resulting in compounds of the formula XIV, wherein
A, B ^PR , D, G, L, R⁵, R⁶, S¹, S², S³, X, X ′, X ′ ′, Y and n are as defined above;
h₁) the protective groups S¹, S², and S³ and the protective groups at B ^{PR are} split off by known processes;
and optionally introducing the groups Q and Q 'by methods known to those skilled in the art, and optionally cyclizing the compounds obtained, resulting in compounds of the formula I. 6. A process for the preparation of the compounds of the formula I according to claims 1 to 4, in which n is 1 to 100, characterized in that in the compounds of the formulas XV and XVI, wherein
A, B ^PR , D, G, L, R⁵, R⁶, S¹, S², S³, X, X ′, X ′ ′ and Y are as defined above,
o and p are independently zero to 50 and o + p + 1 = n;
a₂) n splits off the protective group S¹ from the compounds of the formula XV as described under e₁),
b₂) in the compounds of formula XVI splits off the protective group S³ as described under d₁) and
c₂) couples the resulting compounds to one another as described under f₁), resulting in compounds of the formula XIV, wherein
A, B ^PR , D, G, L, R⁵, R⁶, S¹, S², S³, X, X ′, X ′ ′, Y and n are as defined above,
d₂) and this as described under h₁) to compounds of formula I. 7. A process for the preparation of the compounds of formula I according to claims 1 to 4, characterized in that
a₃) compounds of the formula X, wherein
A, B ^PR , D, G, L, R⁵, R⁶, S¹, S², X, X ′, X ′ ′ and Y are as defined above,
coupled to a solid support via a SPACER using known processes, to give compounds of the formula XVII, wherein
A, B ^PR , D, G, L, R⁵, R⁶, S¹, S², X, X ′, X ′ ′ and Y are as defined above,
SS stands for a solid support suitable for solid phase synthesis, and
SPACER stands for a group which can be split off from the support after synthesis has taken place, as are known to the person skilled in the art, or SPACER stands for bisfunctional conjugate molecules Q which are linked to the solid support via known splitable groups;
b₃) from compounds of the formula XVII, wherein
A, B ^PR , D, G, L, R⁵, R⁶, S¹, S², SS, SPACER, X, X ′, X ′ ′ and Y are as defined above,
splits off the protective group S¹ as described under e₁);
c₃) the resulting compound with compounds of the formula X, wherein
A, B ^PR , D, G, L, R⁵, R⁶, S¹, S², X ′, X ′ ′ and Y are as defined above, as described under f₁);
d₃) steps b₃) and c₃) are repeated until the desired chain length;
e₃) optionally conjugates Q 'coupled by known methods;
f₃) the compounds produced in this way are split off from the solid support by known processes, and the protective groups are described as in step h₁), it being possible for the protective groups to be split off from the support before the cleavage. 8. Use of the compounds of formula I according to claims 1 to 4 as inhibitors of gene expression. 9. Use of the compounds of formula I according to claims 1 to 4 as a diagnostic, for the treatment of diseases caused by viruses caused by integrins or cell-cell adhesion receptors or caused by factors such as TNF alpha, used to treat cancer or the restenosis.