EP1204742A2

EP1204742A2 - Oligonucleotides for inhibiting the expression of human eg5

Info

Publication number: EP1204742A2
Application number: EP00953119A
Authority: EP
Inventors: Eugen Uhlmann; Beate Greiner; Eberhard Unger; Gislinde Gothe; Marc Schwerdel
Original assignee: Aventis Pharma Deutschland GmbH
Current assignee: Sanofi Aventis Deutschland GmbH
Priority date: 1999-07-28
Filing date: 2000-07-21
Publication date: 2002-05-15
Also published as: KR20020033744A; SK1162002A3; ZA200200655B; US6472521B1; MXPA02000817A; YU91901A; BR0013180A; CN1367828A; NO20020365D0; CZ2002324A3; AR024953A1; WO2001007602A3; WO2001007602A2; HRP20020075A2; HUP0202794A3; AU6568200A; HUP0202794A2; HK1048337A1; NZ516839A; TR200200201T2

Abstract

The invention relates to an oligonucleotide or one of its derivatives that is characterized by a sequence that corresponds to a nucleotide sequence that encodes a defined part of human eg5 or a mutated form thereof. The invention further relates to a method of producing said oligonucleotide and to its use.

Description

description

Oligonucleotides to inhibit the expression of human eg5

The present invention relates to an oligonucleotide or one of its derivatives having a sequence which corresponds to a specific part of a nucleic acid sequence which encodes human eg5 or a mutated form thereof, and the invention further relates to a method for producing the oligonucleotide and its use.

During mitosis, a spindle device based on microtubules helps to evenly distribute the duplicated chromosomes among the daughter cells. Motor proteins related to kinesin make up part of the forces required to assemble the spindle and split the chromosomes. The formation of a bipolar mitotic spindle requires the activity of many different motor proteins. One of the human motor proteins related to kinesin is human eg5, which interacts with the mitotic centrosomes and has been shown to be essential for the formation of the bipolar spindle (Blangy et al., Celi (1995) 83, 1159). Microinjection of specific anti-human eg5 antibodies blocks centrosome migration and causes cells to stop mitosis.

Another way to block the formation of a bipolar spindle would be to inhibit eg5 expression. One way to specifically inhibit eg5 expression is to use antisense oligonucleotides, which may have been modified to improve their properties (E. Uhimann and A. Peyman, Chemical Reviews 90, 543 (1990); S. Agrawal, TIBTECH 1996, 376). Antisense oligonucleotides are believed to bind to specific sequences of the mRNA, which leads to degradation of the mRNA and / or inhibition of protein synthesis.

The present invention relates to an oligonucleotide or one of its derivatives which corresponds to part of the eg5 coding sequence - preferably human eg5 or the eg5 of a pathogen, for example Plasmodium falciparum (malaria). The oligonucleotide preferably corresponds to 8 to 100 nucleotides, particularly preferably 8 to 20 nucleotides of the eg5 sequence. The oligonucleotide or the derivative thereof binds to said sequence and inhibits the formation of the eg5 protein. The human eg5 sequence has been published (Blangy et al., Cell (1995) 83, 1159). SEQ ID NO .: 20 shows an example of a sequence encoding human eg5. SEQ ID NO .: 21 shows an example of a Plasmodium falciparum eg 5 sequence.

The oligonucleotide preferably has a sequence that is part of a

Nucleic acid encoding human eg5 or Plasmodium falciparum eg5. The term "corresponds" means that the base sequence of the oligonucleotide is complementary to a part of a nucleic acid sequence which encodes eg5 (eg gene, cDNA, mRNA), which allows the oligonucleotide with the "sense part" of the nucleic acid encoding the egδ to hybridize (bind to it). For this

Reason it is called "antisense oligonucleotide". In a preferred embodiment of the invention, the oligonucleotide is therefore an antisense oligonucleotide. In a further preferred embodiment of the invention, the oligonucleotide is a ribozyme. A ribozyme is a catalytic nucleic acid that cleaves mRNA. The ribozyme is preferably selected from the group of the hammerhead ribozymes (Vaish et al., Nucleic Acids Res. (1998) 26, 5237).

An oligonucleotide according to the invention binds to a part of the eg5 mRNA suitable for hybridization and inhibits the formation of the eg5 protein. Suitable for binding to eg5 mRNA and for inhibiting expression

Oligonucleotides are e.g. targeting the translation starter region of eg5.

The portion of the eg5 coding nucleic acid sequence corresponding to the oligonucleotide has a length of 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 or more nucleotides, and the oligonucleotide preferably corresponds to a length of 12 nucleotides or 19 nucleotides of an eg5 coding sequence. An oligonucleotide according to the invention therefore has a length of 10 (10mer), 11 (11mer), 12 (12mer), 13 (13mer), 14 (14mer), 15 (15mer), 16 (16mer), 17 (17mer), 18 (18mer) or 19 (19mer) nucleotides.

In a preferred embodiment of the invention, the oligonucleotide has a length of 12 or 19 nucleotides; such oligonucleotides can, for example, one of the sequences SEQ ID NO. 1, SEQ ID NO. 2, SEQ ID NO. 3, SEQ ID NO. 4, SEQ ID NO. 5, SEQ ID NO. 6, SEQ ID NO. 7, SEQ ID NO. 8, SEQ ID NO. 9 or a part thereof, wherein SEQ ID NO. 1: 3-'CTTAAGGCAGTACCGCAGC-5 '; 5'CGACGCCATGACGGAATTC-3 'SEQ ID NO. 2: 3'-ACCACTCTACGTCTGGTAA-5 '; 5'-AATGGTCTGCATCTCACCA-3 'SEQ ID NO. 3: 3'-GGCAGTACCGCAGCGTCGG-5 '; 5'-GGCTGCGACGCCATGACGG-3 'SEQ ID NO. 4: 3'-CTTAAGGCAGTA-5 '; 5'-ATGACGGAATTC-3 'SEQ ID NO. 5: 3'-TAAGGCAGTACC-5 '; 5'-CCATGACGGAAT-3 'SEQ ID NO. 6: 3'-GGCAGTACCGCA-5 '; 5'-ACGCCATGACGG-3 'SEQ ID NO. 7: 3'-AGTACCGCAGCG-5 '; S'-GCGACGCCATGA-S "SEQ ID NO.8: 3'-CCGCAGCGTCGG-5 '; 5'-GGCTGCGACGCC-3' SEQ ID NO.9: 3'-GCAGCGTCGGTT-5 '; 5'-TTGGCTGCGACG-3' ,

It is very particularly preferred that the oligonucleotide is modified to improve its properties; e.g. to increase its resistance to nucleases or to make it resistant to nucleases, to improve its binding affinity for a complementary eg5-encoding nucleic acid, e.g. mRNA, or to increase its uptake into the cell.

The present invention therefore preferably relates to an oligonucleotide which has a specific sequence as given above and which, in addition, compared to a "natural" DNA which is composed of the "natural" nucleosides deoxyadenosine (adenine + β-D-2 ^' deoxyribose) , Deoxyguanosine (guanine + ß-D-2 ^' -deoxyribose), deoxycytidine (cytosine + ß-D-2 ^' -deoxyribose) and thymidine (thymine + ß-D-2 ^' -deoxyribose), which are connected by phosphodiester bridges between the nucleosides are, has one or more chemical modifications. The oligonucleotides can contain one or more modifications of the same type and / or modifications of a different type; each type of modification can be made independently of the others be known for the modification of oligonucleotides known modification types.

The invention also relates to derivatives of oligonucleotides, for example their salts, in particular their physiologically tolerable salts. Salts and physiologically acceptable salts are e.g. described in Remington's Pharmaceuticals Science (1985) Mack Publishing Company, Easton, PA (page 1418). Derivatives also refer to modified oligonucleotides that contain one or more modifications (e.g. at certain nucleotide positions and / or at certain internucleoside bridges), oligonucleotide analogs (e.g. polyamide nucleic acids

(PNAs), phosphomonoester nucleic acids (PHONAs = PMENAs), oligonucleotide chimers (e.g. consisting of a DNA and a PNA part or consisting of a DNA and a PHONA part)). Derivatives also refer to oligonucleotides corresponding to alleles and / or mutated forms (mutants) of normal or natural eg5, e.g. Alleles and / or mutants of human eg5 (e.g. related to SEQ ID NO. 20) and alleles and / or mutants of Plasmodium falciparum eg5 (e.g. related to SEQ ID NO. 21).

Examples of chemical modifications are known to the person skilled in the art and are described, for example, in E. Uhlmann and A. Peyman, Chemical Reviews 90 (1990) 543 and "Protocols for Oligonucleotides and Analogs" Synthesis and Properties & Synthesis and Analytical Techniques, S. Agrawal, ed., Humana Press, Totowa, USA 1993 and ST Crooke, F. Bennet, Ann. Rev. Pharmacol. Toxicol. 36 (1996) 107-129; J. Hunziker and C. Leuman (1995) Mod. Synt. Methods, 7, 331-417.

In comparison to natural DNA, for example, a phosphodiester bridge between nucleosides, a β-D-2 ^' deoxyribose unit and / or a natural nucleoside base (adenine, guanine, cytosine, thymine) can be modified or exchanged. An oligonucleotide according to the invention can have one or more of these modifications, each modification compared to an oligonucleotide of the same sequence consisting of natural DNA on a specific phosphodiester internucleoside bridge and / or on a specific β- D-2 ^' deoxyribose unit and / or at a specific natural nucleus base position.

For example, the invention relates to an oligonucleotide which contains one or more modifications, each modification being selected independently from the following list: a) the exchange of a phosphodiester bridge between nucleosides which are located at the 3 ^' and / or at the 5 ^' end of one Nucleoside is through a modified internucleoside bridge, b) the exchange of a phosphodiester bridge located at the 3 ^' and / or the 5 ^' end of a nucleoside with a dephospho bridge, c) the exchange of a sugar phosphate unit from the sugar phosphate skeleton with another unit, d ) the exchange of a β-D-2 ^' -deoxyribose unit by a modified sugar unit, e) the exchange of a natural nucleoside base by a modified nucleoside base, f) the connection to a molecule which influences the properties of the oligonucleotide, g) the connection to a 2'5'-linked oligoadenylate or a derivative thereof, optionally via a suitable link er, and h) the introduction of a 3 ^' -3 ^' and / or a 5 ^' -5 ^' inversion at the 3 ^' and / or at the 5 ^' end of the oligonucleotide.

More precise examples of the chemical modifications of an oligonucleotide are a) the exchange of a phosphodiester bridge between nucleosides, which is located at the 3 ^' and / or at the 5 ^' end of a nucleoside, with a modified internucleoside bridge, the modified internucleoside bridge being made, for example, from phosphorothioate , Phosphorodithioat-, NR ¹ R ^r -phosphoramidate-, boranophosphate-, phosphate- (Cι-C ₂ ι) -0-alkyl ester-, phosphate - [(C ₆ -Cι ₂ ) aryl - ((C ₁ -C ₂ ι) -0-alkyl] esters, (CrC ₈ ) alkylphosphonate and / or (C ₆ -C ₂ ) arylphosphonate bridges and (C ₇ -C ι ₂ ) -α- hydroxymethylaryl (known for example from WO 95/01363 ) is selected, with (C ₆ -Cι ₂ ) - Aryl, (C ₆ -C ₂ o) aryl and (C ₆ -Cι ₄ ) aryl optionally by halogen, alkyl, alkoxy,

Nitro, cyano are substituted, and wherein R ¹ and R ¹ independently of one another are hydrogen, (C ₈ -C) alkyl, (C6-C ₂ o) aryl,

(C ₆ -C ₄ ) aryl- (-C ₈ ) alkyl, preferably hydrogen, (CC ₈ ) alkyl, preferably (C 1 -C ₄ ) alkyl and / or methoxyethyl, or

R ¹ and R ¹ together with the nitrogen atom carrying them form a 5- to 6-glιedrιgen heterocyclic ring which additionally contains a further heteroatom from the

Can contain groups O, S and N,

b) the replacement of a phosphodiester bridge located at the 3 ^' and / or the 5 ^' end of a nucleoside by a dephospho bridge (dephospho bridges are described, for example, in Uhlmann, E and Peyman, A in "Methods in Molecular Biology", volume 20, "Protocols for Oligonucleotides and Analogs", S Agrawal, ed., Humana Press, Totowa 1993, chapter 16, 355ff.), A dephosphobrucke for example formacetal, 3'-thioformacetal, methylhydroxylamine, oxime, methylenedimethylhydrazo, dimethylene sulfone and / or is a silyl group,

c) the exchange of a sugar phosphate unit (ß-D-2'-deoxyπbose and the phosphodiester bridge between the nucleosides together form a sugar phosphate unit) from the sugar phosphate framework (the sugar phosphate framework consists of sugar phosphate units) by another unit, the other unit being used, for example, for assembly - a "Morpholιnodeπvatιv" oligomer is suitable (as for example in EP

Stirchak et al., Nucleic Aαds Res 17 (1989) 6129), that is, for example, the exchange by a morpho-derivative unit, a polyamide nucleic acid ("PNA") (as described, for example, in PE Nielsen et al. Bioconj Chem 5 (1994) 3 and described in EP 0672677 A2) is suitable, that is, for example, the exchange by a PNA framework, for example by

2-Amιnoethylglycιn, a phosphonic acid monoester nucleic acid ("PHONA") (such as in Peyman et al, Angew Chem Int Ed Eπgl 35 (1996) 2632-2638 and in EP 0739898 A2) is suitable; this means, for example, the exchange by a PHONA scaffolding unit; d) the replacement of a β-D-2 ^' deoxyribose unit by a modified sugar unit, the modified sugar unit being composed, for example, of β-dribose, α-D-2'-deoxyribose, L-2'-deoxyribose, 2'-F- 2'-deoxyribose, 2'-0- (Cι-C ₆ ) alkyl ribose, the preferred 2'-0- (Cι-C ₆ ) alkyl ribose being 2'-0-methylribose, 2'-0- (C ₂ -C ₆ ) alkenyl ribose, 2 '- [0- (C ₁ -C ₆ ) alkyl-0- (C ₁ -C ₆ ) alkyl] ribose, 2'-NH ₂ -2'-deoxyribose, ß -D-xylo-furanose, α-arabinofuranose, 2,4-dideoxy-β-D-erythro-hexo-pyranose and carbocyclic (for example described in Froehler, J. Am. Chem. Soc. 114 (1992) 8320) and / or open-chain sugar analogues (for example in

Vandendriessche et al., Tetrahedron 49 (1993) 7223) and / or bicydosugar analogs (e.g. described in M. Tarkov et al., Helv. Chim. Acta 76 (1993) 481);

e) the replacement of a natural nucleoside base by a modified nucleoside base, the modified nucleoside base consisting, for example, of uracil, hypoxanthine, 5- (hydroxymethyl) uracil, N ² -dimethylguanosine, pseudouracil, 5- (hydroxymethyl) uracil, 5-aminouracil, dihydrouracil, 5-fluorouracil, 5-fluorocytosine, 5-chlorouracil, 5-chlorocytosine, 5-bromouracil, 5-bromocytosine, 2,4-diaminopurine, 8-azapurine, a substituted 7-deazapurine, preferably 7-deaza-7-substituted and / or 7-deaza-8-substituted purine or other modifications of natural nucleoside bases is selected (modified nucleoside bases are described, for example, in EP 0 710 667 A2 and EP 0 680 969 A2);

f) the connection to a molecule which influences the properties of the oligonucleotide, the connection of the oligonucleotide to one or more molecules which influence the properties of the oligonucleotide (favorably) (for example the ability of the oligonucleotide to penetrate the cell membrane or in to penetrate a cell, the stability towards nucleases, the affinity for an eg5 coding target sequence, the pharmacokinetics of the oligonucleotide, the ability of an antisense oligonucleotide / ribozyme or a molecule conjugated with the oligonucleotide, to attack the respective eg5 coding target sequence, eg the ability to attack Binding and / or cross-linking when the oligonucleotide hybridizes with the eg5-encoding target sequence), polyiysin, intercalating agents such as pyrene, acridine, phenazine or phenanthridine, fluorescent agents such as fluorescein, crosslinkers such as psoralen or azidoproflavin, lipophilic molecules such as (Cι.) as examples of molecules which can be bound to an oligonucleotide ₂ -C ₂ o) alkyl, lipids such as 1, 2-dihexadecyl-rac-glycerol, steroids such as cholesterol or testosterone, vitamins such as vitamin E, poly- or oligoethylene glycol, preferably via a phosphate group (eg triethylene glycol phosphate, hexaethylene glycol phosphate) to the oligonucleotide bound, (-C ₂ -C ₁₈ ) alkyl phosphate diester and / or 0-CH ₂ -CH (OH) -0- (C ₁₂ -C ₈ ) - alkyl come into question, these molecules can at the 5 'end and / or be inserted at the 3 'end and / or within the sequence, for example on a nucleoside base to form an oligonucleotide conjugate; Methods for producing an oligonucleotide conjugate are known to the person skilled in the art and are described, for example, in Uhlmann, E. & Peyman, A., Chem. Rev. 90 (1990) 543, M. Manoharan in "Antisense Research and Applications", Crooke and Lebleu, ed. , CRC Press, Boca Raton, 1993, Chapter 17, pp. 303ff. and EP-A 0 552 766;

g) the connection to a 2'5'-linked oligoadenylate, preferably via a suitable linker molecule, the 2'-5'-linked oligoadenylate consisting, for example, of 2'5'-linked triadenylate, 2'5'-linked tetraadenylate .

2'5'-linked pentaadenylate, 2'5'-linked hexaadenyltate or 2'5'-linked heptaadenylate molecules and their derivatives is selected, a 2 ^' 5 ^' -linked oligoadenylate derivative being, for example, cordycepin (2'5 ' -bound 3'-deoxyadenylate) and where, for example, triethylene glycol is a suitable linker and where the 5 'end of the 2'5'-linked oligoadenylate must carry a phosphate, diphosphate or triphosphate radical in which one or more oxygen atoms are used Example can be replaced by sulfur atoms, the substitution by a phosphate or thiophosphate residue is preferred; and

h) the introduction of a 3 ^' -3 ^' and / or a 5 ^' -5 ^' inversion at the 3 'and / or at the 5' end of the oligonucleotide, this type of chemical modification corresponding to the Known to those skilled in the art and described for example in M. Koga et al., J. Org. Chem. 56 (1991) 3757, EP 0 464 638 and EP 0 593 901.

The replacement of a sugar phosphate unit from the sugar phosphate framework by another unit, e.g. a PNA scaffolding unit or a PHONA

The scaffold unit can preferably be the replacement of a nucleotide by, for example, a PNA unit or a PHONA unit which already contains natural nucleoside bases and / or modified nucleoside bases, for example one of the modified nucleoside bases from the group uracil, hypoxanthine, 5- (hydroxymethyl) uracil, N ² - dimethylguanosine, pseudouracil, 5- (hydroxymethyl) uracii, 5-aminouracil, pseudouracil, dihydrouracil, 5-fluorouracil, 5-fluorocytosine, 5-chlorouracil, 5-chlorocytosine, 5-bromouracil, 5-bromocytosine, 2, 4-diaminopurine, 8-azapurine, a substituted 7-deazapurine, preferably 7-deaza-7-substituted and / or 7-deaza-8-substituted purine or other modifications of a natural nucleoside base (modified nucleotide bases are described, for example, in EP 0 710 667 A2 and EP 0 680 969 A2).

The oligonucleotide modifications described in EP 0 710 667 A2, EP 0 680 969 A2, EP 0 464 638, EP 0 593 901, WO 95/01363, EP 0 672 677 A2, EP 0 739 898 A2 and EP 0 552 766 explicit reference is hereby made.

In a particular embodiment of the invention, one or more phosphodiester bridges between the nucleosides within the oligonucleotide sequence are modified; are preferably one or more phosphodiester bridges between the nucleosides by phosphorothioate bridges between the nucleosides and / or (C ₆ -Ci ₂ ) arylphosphonate bridges between the nucleosides, preferably by α-hydroxybenzylphosphonate bridges in which the benzyl group is preferably substituted, for example by nitro, methyl, Halogen, replaced. In a phosphorothioate oligonucleotide only, all phosphodiester bridges between the nucleosides are modified by phosphorothioate. The invention preferably relates to an oligonucleotide in which not all phosphodiester bridges between the nucleosides are modified uniformly with phosphorothioate (phosphorothioate bridges between the nucleosides). Preferably at least one internucleoside bridge has a different type of modification or it is not modified. The invention relates in particular to an oligonucleotide which additionally contains at least one other type of modification. In a further particular embodiment of the invention, one or more nucleosides (β-D-2 ^' deoxyribose and / or nucleoside base) are modified within the oligonucleotide sequence; the β-D-2 ^' deoxyribose is preferably replaced by 2'-0- (CrC ₆ ) -alkyl ribose, preferably by 2 ^' -0-methyl ribose, and / or the nucleoside base is 8-azapurine, 7-deaza-7 -substituted purine and / or 7-deaza-8-substituted purine (purine: adenine, guanine). The invention preferably relates to an oligonucleotide in which not all nucleosides are uniformly modified. The invention preferably relates to an oligonucleotide which additionally contains at least one other type of modification.

In a further particular embodiment of the invention, one or more sugar phosphate units are removed from the sugar-phosphate framework by PNA-

Framework units, preferably replaced by 2-aminoethylglycine units. The exchanged sugar phosphate units are preferably connected to one another at least to a certain degree. The invention preferably relates to an oligonucleotide in which not all sugar phosphate units are exchanged uniformly. The invention particularly relates to chimeric oligonucleotides, e.g. those composed of one or more PNA parts and one or more DNA parts. Possible examples of such chimeric oligonucleotides are the following modification patterns, which do not serve to restrict the invention: DNA-PNA, PNA-DNA, DNA-PNA-DNA, PNA-DNA-PNA, DNA-PNA-DNA-PNA, PNA- DNA-PNA-DNA. Similar patterns would be out

DNA parts and PHONA parts composed of chimeric molecules possible, for example DNA-PHONA, PHONA -DNA, DNA-PHONA -DNA, PHONA -DNA- PHONA, DNA- PHONA -DNA- PHONA, PHONA -DNA- PHONA -DNA. In addition, chimeric molecules from three different parts such as DNA part (s), PHONA part (s) and PNA part (s) are of course also possible. The invention preferably relates to an oligonucleotide which additionally contains at least one other type of modification. In a further particular embodiment of the invention, the 3 ^' end and / or the 5 ^' end of the oligonucleotide is connected to a (C ₂ -C ₁₈ ) allyl residue, preferably a C ₆ alkyl residue, a triethylene glycol residue or a hexaethylene glycol residue - These residues are preferably bound to the oligonucleotide via a phosphate group. The invention preferably relates to an oligonucleotide in which both ends (the 3 ^' and the 5 ^' end) are not modified (equally). The invention preferably relates to an oligonucleotide which additionally contains at least one other type of modification.

In a preferred embodiment of the invention, only certain positions within an oligonucleotide sequence are modified (e.g. partially modified oligonucleotide). In some publications, partially modified oligonucleotides are also referred to as minimally modified oligonucleotides. A modification can be located in certain positions within the sequence (with certain nucleotides, with certain nucleosides, with certain nucleoside bases, with certain internucleoside bridges).

In a particular embodiment of the invention, a partially modified oligonucleotide is made by only some of the phosphodiester bridges through modified internucleoside bridges, e.g. Phosphorothioate bridges and / or α-hydroxybenzylphosphonate bridges can be replaced. The invention particularly includes those oligonucleotides that are only modified to a certain degree.

In particular, the invention relates to an oligonucleotide in which 1 to 5 terminal nucleotide units at the δ 'end and / or at the 3' end by modifying internucleoside bridges which are located at the 5 'and / or at the 3' end of the corresponding nucleoside are protected, preferably by exchanging the phosphodiester bridges between the nucleosides by phosphorothioate bridges and / or α-hydroxybenzylphosphonate bridges. The 1 to 5 terminal nucleotide units at the 3 'end of the oligonucleotide are very particularly preferably protected by modified internucleoside bridges which are located at the 5' and / or at the 3 'end of the corresponding nucleosides. If necessary, the 1 to 5 Terminal nucleotide units at the 5 'end of the oligonucleotide are additionally protected by modified internucleoside bridges which are located at the 5' and / or at the 3 'end of the corresponding nucleoside. The oligonucleotide can optionally contain additional modifications at other positions. The invention further relates to an oligonucleotide in which at least one internal pyrimidine nucleoside and / or an internucleoside bridge located at the 5 'end and / or at the 3' end of this pyrimidine nucleoside (a nucleoside with a pyrimidine base such as cytosine, uracil, thymine) is modified is, preferably by exchanging the phosphodiester bridge (s) between the nucleosides by one / more phosphorothioate bridge (s) and / or one / more α-hydroxybenzylphosphonate bridge (s).

In a preferred embodiment of the invention, 1 to 5 terminal nucleotide units at the 5 'end and / or at the 3' end of the oligonucleotide are by modifying internucleoside bridges which are located at the 5 'and / or at the 3' end of the corresponding nucleoside, protected, in addition at least one internal pyrimidine nucleoside and / or an internucleoside bridge located at the 5 'end of this pyrimidine nucleoside and / or at the 3' end of this pyrimidine nucleoside is modified.

The principle of partially modified oligonucleotides is described, for example, in A. Peyman, E. Uhlmann, Biol. Chem. Hoppe-Seyler, 377 (1996) 67-70 and in EP 0 653 439. Reference is hereby expressly made to these documents. In this case 1-5 terminal nucleotide units are protected at the 5 'end and / or at the 3' end, for example the phosphodiester bridges between the nucleosides which are at the 3 'and / or at the 5' end of the corresponding nucleosides , for example, exchanged for phosphorothioate bridges between nucleosides. In addition, at least one internal pyrimidine nucleoside position (or nucleotide position) is preferably modified; the 3 'and / or the 5' internucleoside bridge (s) of a pyrimidine nucleoside is / are preferably modified / exchanged, for example by one / more phosphorothioate bridge (s) between the nucleosides. Partially modified oligonucleotides have particularly advantageous properties; for example, they are particularly stable to nucleases, and only minimally modified. They also have a considerably reduced tendency to non-antisense effects, which are often associated with the use of phosphorothioate-only oligonucleotides (Stein and Krieg (1994) Antisense Res. Dev. 4, 67). Partially modified oligonucleotides also show a higher binding affinity than phosphorothioates only.

The invention particularly relates to partially / minimally modified oligonucleotides.

SEQ ID NO. 10: 3-'C ^* T ^* T ^* AAGGC ^* AGT ^* AC ^* CG ^* CAG ^* C ~ 5 \ (K3) 5'-CGAC ^* G * C ^* C ^* A ^* TGA * CGGAA ^* T * T * C -3 ';

SEQ ID NO. 11: 3'-A ^* C * C * AC ^* TC ^* TAC ^* GT * C ^* TGG ^* TA ^* A-5 ', (K4)

5'-A ^* AT ^* GGT ^* C ^* TG ^* CAT ^* CT ^* CA * C ^* C ^* A-3 '; SEQ ID NO. 12: 3'-G ^* G ^* C ^* AG ^* TAC ^* CGC ^* AG ^* CGT ^* CG ^* G-5 ', (K6) 5'-G * GC ^* TGC ^* GA ^* CGC * CAT ^* GA * C ^* G ^* G-3 "; SEQ ID NO. 13: 3 ^* -C ^* T ^* T ^* AAGG ^* CAG ^* T ^* A-5 ',

5'-A * T ^* GAC ^* GGAA ^* T ^* T ^* C-3 '; SEQ ID NO. 14: 3'-T ^* A ^* AGGC ^* AG ^* TA ^* C ^* C-5 ', 5'-C ^* C ^* AT ^* GA ^* CGGA ^* A * T-3'; SEQ ID NO. 15: 3'-G ^* G ^* CAG ^* TAC ^* C ^* GC ^* A-5 ', 5'-A ^* CG ^* C ^* CAT ^* GAC ^* G * G-3';

SEQ ID NO. 16: 3'-A ^* G ^* TAC ^* CG ^* CAG ^* C ^* G-5 ^* , 5'-G * C * GAC ^* GC ^* CAT ^* G ^* A-3 '; SEQ ID NO. 17: 3'-C * C ^* G ^* CAG ^* CGT ^* CG ^* G-5 ', 5'-G * GC ^* TGC ^* GAC ^* G ^* C ^* C-3 "; SEQ ID NO. 18 3'- G * C ^* AGC ^* GT ^* CGG ^* T ^* T-5 ',

5'-T * T * GGC ^* TGC ^* GA ^* C * G-3 \

where "*" denotes the site of an internucleoside bridge modification; "*" is preferably a phosphorothioate internucleoside bridge.

Another example of a particular embodiment of the invention relates to a partially modified oligonucleotide in which a nucleoside is modified, for example a modification of a nucleoside base and / or a modification of a β-D-2 ^' deoxyribose unit. A ß-D-2 ^' -deoxyribose is preferably exchanged for 2'-O- (-C-C ₆ ) alkyliribose, the exchange for is very particularly preferred 2'-0-methylribose (exchange of a β-D-2 ^' deoxyribonucleoside for a 2 ^' -0-methylribonucleoside).

In addition to one type of modification, the oligonucleotide can also have other types of modification according to the invention.

In a further embodiment of the invention, the oligonucleotide therefore contains modified internucleoside bridges at certain positions and additionally modifications of a nucleoside at certain positions, preferably exchange of β-D-2 ^' deoxyribose. In a preferred embodiment of the invention, the internucleoside modification is the exchange of a phosphodiester bridge for a phosphorothioate bridge, and the modification of the β-D-2 ^" deoxyribose is the exchange for 2 ^' -0-methylribose; in this case the oligonucleotide is a chimeric oligonucleotide, which is composed of modified and unmodified DNA and RNA parts - which are the

Contain 2'-0-methyl ribonucleosides and ß-D-2 ^' deoxyribonucleosides and phosphorus diester and phosphorothioate bridges between nucleosides.

A further preferred embodiment of the invention relates to an oligonucleotide which has one or more (C ₂ -C ₈ ) alkyl residues, preferably one

C ₁₆ alkyl radical at its 3 'and / or its 5' end. A (-C ₂ -C ₁₈ ) -alkyl radical can be bound, for example, as a phosphodiester, as described in EP 0 552 766 A2 (EP 0 552 766 A2 is hereby expressly incorporated by reference), or as a 3'-phosphodiester of 0-CH ₂ -CH (OH) -0- (-C ₂ -C ₁₈ ) alkyl. An oligonucleotide in which a C ₆ -alkyl radical is bonded to the 3 'and / or 5' end is preferred.

The invention also relates to an oligonucleotide in which the 3 'and / or the 5' end is connected to an oligoethylene glycol residue, preferably a triethylene glycol or a hexaethylene glycol, very particularly preferably via a phosphodiester (tri- or hexaethylene glycol phosphate ester). Such an oligonucleotide can of course also contain further modifications. In a further particular embodiment of the invention, the oligonucleotide is linked via a linker to a 2'5'-linked oligoadenylate 5 '- (thio) phosphate. The linker can be, for example, an oligoethylene glycol phosphate, preferably a triethylene glycol phosphate, tetraethylene glycol phosphate or hexaethylene glycol phosphate residue. The 2'5'-bound oligoadenylate is preferably bound via its 2'-end as a tetra- or as a pentaadenylate, the 5'-hydroxy function of which is substituted by a phosphate or thiophosphate radical. It is known that 2'5'-oligoadenylate induces RNase L to cleave the target mRNA (Torrence et al., Proc. Natl. Acad. Sei. USA (1993) 90, 1300). The 2'5'-oligoadenylate serves to activate ribonuclease L (RNase L), which then degrades the eg5 mRNA. Instead of a 2'5'-linked adenylate, it is also possible to introduce, for example, a 2'5'-linked 3'-deoxyadenylate derived from the nucleoside analogue cordycepin. In this case, the part of the oligonucleotide which is complementary to the target nucleic acid is preferably modified at certain positions by 2'-0- (C ₁ -C ₆ ) -alkyl ribonucleoside (preferably 2'-0-methylribonucleoside) or by PNA.

Another preferred embodiment of the invention relates to the replacement of one or more natural nucleoside bases by unnatural or modified nucleoside bases, preferably by 8-azapurines and / or 7-deaza-7-substituted purines and / or 7-deaza-8-substituted ones Purines, such as in EP 0 171 066 and EP 0 680 969.

In a further preferred embodiment of the invention, the nucleoside can have 3'3 'and / or 5'5' inversions at the 3 'and / or 5' end, as for example in EP 0 464 638 and EP 0 593 901 described.

A further preferred embodiment of the invention relates to the exchange of one or more phosphodiester bridges for α-hydroxybenzylphosphonate bridges, as described in WO 95/01363. In a further preferred embodiment of the invention, the oligonucleotide contains a modification of the sugar phosphate structure, preferably by PNA units.

Other modification patterns are also possible, e.g. DNA-PNA-DNA, PNA-DNA. Comparable modification patterns are also possible for PHONA / DNA chimers. These modification patterns can be combined with any other type of modification, and of course similar modification patterns are also possible for other oligonucleotides according to the invention.

The above-mentioned specific oligonucleotides - specific sequence, specific modification type / specific modification types at specific positions (specific "modification pattern") are only examples of different embodiments of the invention. The invention is not restricted by these specific oligonucleotides. Other combinations of sequence and modification pattern are also possible.

An oligonucleotide according to the invention specifically inhibits the expression of the target protein (i.e. eg5) or the target sequence (a nucleic acid which codes for eg5, preferably eg5 mRNA). An oligonucleotide according to the invention preferably specifically inhibits the expression of eg5. This results in a lowering of the eg5 protein concentration compared to an untreated expression. The specificity can be demonstrated, for example, by determining the effect of an oligonucleotide according to the invention on eg5 expression in comparison to the effect of the same oligonucleotide on beta actin expression on the mRNA and / or the protein level. After treatment with an oligonucleotide according to the invention, only the eg5 mRNA and / or the eg5 protein concentration was reduced, while e.g. beta-actin (a household protein) mRNA and / or beta-actin protein concentration remained unchanged.

An oligonucleotide according to the invention is preferably capable of and / or has the ability to effectively inhibit the expression of eg5 in human cells Ability to inhibit the growth of tumors in vertebrates. An oligonucleotide according to the invention preferably reduces the eg5 mRNA and / or protein concentration in tumors of treated individuals compared to untreated individuals. An oligonucleotide according to the invention preferably reduces the size of a tumor in a vertebrate, for example in mice, in comparison to untreated mice or in comparison to the size of the tumor determined in the same animal before treatment.

The invention also relates to a method for producing a nucleotide according to the invention. One manufacturing process involves chemical synthesis of the oligonucleotide. Chemical synthesis is preferably carried out using a standard method known for use in the synthesis of oligonucleotides, for example the phosphoramidite method according to Caruthers (1983) Tetrahedron Letters 24, 245, the H-phosphonate method (Todd et al. (1957) J. Chem. Soc . 3291) or the phosphotriester method (Sonveaux (1986) Bioorg. Chem. 14, 274; Gait, MJ Oligonucleotide Synthesis, A practical Approach ", IRL Press, Oxford, 1984) or improved or modified methods which are derived from these standard methods. For example, an oligonucleotide according to the invention can be prepared as described in Example 1. An oligonucleotide according to the invention is preferably synthesized on a solid phase by suitably condensing protected monomers (eg nucleosides) to form internucleoside bridges between these monomers for the preparation of an oligonucleotide or a derivative thereof, w or a nucleotide unit with a 3 ^' or a 2 ^' terminal phosphorus (V) group and a free 5 ^' hydroxyl or mercapto grouping with a further nucleotide unit with a phosphorus (III) or a phosphorus (V) grouping in the 3 'position, or their activated derivatives, and optionally using protective groups which can be temporarily introduced into the oligonucleotide to protect other functions and which are removed after synthesis and the one which is split off from the solid phase Oligonucleotide can optionally be converted into a physiologically acceptable salt. To synthesize a modified oligonucleotide, the standard methods are varied to some extent. These are variations known to the person skilled in the art, and they are described, for example, in Agrawal S. "Protocols for oligonucleotides and analogs" (1993, Human Press Inc., Totowa, New Jersey). The preparation of modified oligonucleotides is also described in EP 0 710 667, EP 0 680 969, EP 0464 638, EP 0 593 901, WO 95/01363, EP 0 672 677, EP 0 739 898 and EP 0 552 766. Reference is hereby expressly made to the methods for producing modified oligonucleotides described in the abovementioned documents.

The invention further relates to a method for inhibiting the expression of eg5 and / or for modulating the expression of a nucleic acid encoding eg5, an oligonucleotide according to the invention being brought into contact with a nucleic acid encoding eg5 (eg mRNA, cDNA) and the oligonucleotide encoding this eg5 Nucleic acid is hybridized.

The invention therefore also relates to a method in which the oligonucleotide is brought into contact with a nucleic acid encoding eg5 (eg mRNA; cDNA), for example by inserting the oligonucleotide into a cell using known methods, for example by incubating cells with the above-mentioned oligonucleotide or a formulation thereof - such a formulation may contain uptake improvers such as lipofectin, lipofectamine, cellfectin or polycations (eg polylysine).

For example, an oligonucleotide previously incubated with Cellfectin, for example for 30 minutes at room temperature, is then incubated with a cell for about 5 hours or less to introduce the oligonucleotide into the cell. The invention further relates to the use of the oligonucleotide, preferably as an antisense oligonucleotide (binding of the oligonucleotide to an mRNA coding for eg5) or as a ribozyme (binding to a mRNA coding for eg5 and cleavage of this mRNA). In a further particular embodiment of the invention, the oligonucleotide can be used to induce the cleavage of the mRNA encoding eg5 by RNase H, which leads to a reduced eg5 expression. The invention relates to the use of an oligonucleotide for inhibiting the formation of a bipolar mitotic spindle and thus for inhibiting cell proliferation, in particular tumor growth.

The invention further relates to the use of the oligonucleotide as a medicament and the use of the oligonucleotide for the production of a pharmaceutical preparation. In particular, the oligonucleotide can be used in a pharmaceutical preparation which is used for the prevention and / or for the treatment of diseases which are associated with the expression of eg5 or which can be cured by the inhibition of eg5 expression.

The invention further relates to a pharmaceutical preparation which contains an oligonucleotide and / or its physiologically acceptable salts together with pharmaceutically acceptable carriers or auxiliaries.

The invention relates to a pharmaceutical preparation which contains at least one nucleotide according to the invention which can be used for the treatment of diseases which can be cured by the inhibition of eg5 expression, such as restenosis and cancer.

The invention further relates to a method for producing a pharmaceutical preparation, wherein one or more oligonucleotides according to the invention with physiologically acceptable carriers and optionally additional substances, e.g. optionally mixed with suitable additives and / or auxiliaries.

The invention relates in particular to the use of an oligonucleotide or a pharmaceutical preparation prepared therefrom for the treatment of cancer, for example for the inhibition of tumor growth and tumor metastasis. The oligonucleotide or a pharmaceutical preparation produced therefrom can be used, for example, for the treatment of solid tumors, such as breast cancer, lung cancer, head and neck cancer, brain cancer, stomach cancer, colon cancer, colorectal cancer, esophageal cancer, gastrointestinal cancer, gliatumor, liver cancer, tongue cancer, Neuroblastoma, osteosarcoma, ovarian cancer, pancreatic cancer, prostate cancer, retinoblastoma, Wilms tumor, multiple myeloma can be used, and for the treatment of skin cancer such as melanoma, for the treatment of lymph node tumors and blood cancer. The invention further relates to the use of an oligonucleotide according to the invention or a pharmaceutical preparation prepared therefrom for inhibiting eg5 expression and / or for inhibiting the accumulation of ascitic fluid and pleural effusions in various types of cancer, for example breast cancer, lung cancer, head cancer, neck cancer, brain cancer, abdominal cancer, and colon cancer , colorectal cancer, esophageal cancer, gastrointestinal cancer, gliatumor, liver cancer, tongue cancer, neuroblastoma, osteosarcoma, ovarian cancer, pancreatic cancer, prostate cancer, retinoblastoma, Wilms tumor, multiple myeloma, skin cancer, melanoma and blood cancer. Due to the inhibitory effect on eg5 expression, an oligonucleotide according to the invention or a pharmaceutical composition produced therefrom can improve the quality of life.

The invention further relates to the use of an oligonucleotide or a pharmaceutical preparation thereof, e.g. for the treatment of cancer or for the prevention of tumor metastasis, in combination with other drugs and / or other therapeutic measures, e.g. with known drugs and / or known therapeutic measures, such as those currently used for the treatment of cancer and / or for the prevention of tumor metastasis. A combination with radiation therapy and chemotherapeutic agents such as cisplatin, cyclophosphamide, 5-fluorouracil, adriamycin, daunorubicin or tamoxifen is preferred.

The oligonucleotide and / or its physiologically acceptable salt can be administered to an animal, preferably a mammal and in particular a human being alone, in a mixture with another oligonucleotide (or its physiologically acceptable salt) or in the form of a pharmaceutical preparation which the topical, percutaneous , parenteral or enteral use and as an active ingredient an effective dose of at least one oligonucleotide, in addition to usual pharmaceutically acceptable Carriers and auxiliary substances. Such a pharmaceutical preparation normally contains about 0.1 to 90% by weight of the therapeutically active oligonucleotide / oligonucleotides. The dose can be varied within a wide range and must be adapted to the individual circumstances. Topical use is preferred for treating psoriasis. In the case of cancer, infusions, oral and rectal administration, or nasal administration in an aerosol, preferably in the case of lung cancer, are preferred, while in the case of diabetic retinopathy, topical, intravitreal and oral administration is preferred.

A pharmaceutical preparation can be prepared in a manner known per se (e.g. Remingtons Pharmaceutical Sciences, Mack Publ. Co., Easton, PA (1985)) using pharmaceutically inert inorganic and / or organic carriers. Lactose, corn starch and / or its derivatives, talc, stearic acid and / or its salts, etc., can be used, for example, for the production of pills, tablets, film-coated tablets and hard gelatin capsules. Examples of carriers for soft gelatin capsules and / or suppositories are fats, waxes, semi-solid and liquid polyols, natural and / or hardened oils, etc. Examples of suitable carriers for the preparation of solutions and / or syrups are water, sucrose, invert sugar, glucose, polyols , etc. Suitable carriers for the preparation of injection solutions are water, alcohols, glycerol, polyols, vegetable oils, etc. Suitable carriers for microcapsules, implants and / or rods are mixed polymers of glycolic acid and lactic acid. In addition there are liposome formulations which e.g. in N. Weiner (Drug Develop Ind Pharm 15 (1989) 1523), "Liposome Dermatics" (Springer Verlag 1992) and Hayashi (Gene Therapy 3 (1996) 878). The pharmaceutical composition may also comprise a formulation which increases the oral availability of the oligonucleotide, such as substances for improving intestinal uptake, e.g. Mannitol, urea, bile acid salts such as CDCA (Chenodeoxycholate) (2%).

Dermal application is also possible, for example using ionophoretic methods and / or using electroporation. In addition, lipofectins and other carrier systems, for example those in gene therapy used, used. Systems which allow the introduction of oligonucleotides into eukaryotic cells or into the nucleus of eukaryotic cells in a highly efficient manner are particularly suitable. A pharmaceutical preparation can also consist of two or more different oligonucleotides and / or their physiologically acceptable salts and, in addition to at least one oligonucleotide, of one or more different therapeutic active ingredients. In addition to the active substances and carriers, a pharmaceutical preparation can also contain additives such as fillers, extenders, disintegrants, binders, lubricants, wetting agents, stabilizers, emulsifiers, preservatives, sweeteners,

Contain colors, flavors or flavors, thickeners, diluents or buffer substances, and in addition solvents and / or solubilizers and / or agents for achieving a retarding effect, and also salts for changing the osmotic pressure, coating agents and / or antioxidants.

Examples:

Example 1: Oligonucleotide Synthesis

Oligonucleotides (ON s) were synthesized using an Applied Biosystems 394 DNA synthesizer (Perkin Elmer Applied Biosystems, Inc., Foster City, USA) and standard phosphoramidite chemistry. After coupling, phosphorothioate bonds were introduced by sulfurization using the Beaucage reagent followed by capping with acetic anhydride and N-methylimidazole. After cleavage from the solid phase and final deprotection by treatment with concentrated ammonia, the ONs were purified by polyacrylamide gel electrophoresis. The 2'-O-Mef 7y / -modified ON s were prepared by exchanging the standard phosphoramidites in the corresponding cycle for 2'-0-methylribonucieoside phosphoramidites. All ON s were analyzed by electrospray mass spectroscopy with negative ions (Fisons Bio-Q), which confirmed the calculated mass in all cases. The C16 modified oligonucleotides were synthesized using hexadecyioxy (cyanoethoxy) -N, N-diisopropyl-aminophosphane instead of a standard amidite as phosphitylation reagent in the last step of the oligonucleotide synthesis or starting from a correspondingly derivatized solid phase. The triethylene glycol linker is commercially available from Glen Research Corporation. The 2'-phosphoramidites of adenosine and cordycepin were purchased from Chem. Genes Corporation and Chemogen Corporation, respectively. The introduction of 5'-phosphates or thiophosphate residues was carried out as previously described (Uhlmann and Engels (1986) Tetrahedron Lett. 27, 1023). The PNA-DNA chimers were produced as described in EP 0 672 677.

The oligonucleotides were analyzed by a) analytical gel electrophoresis in 20% acrylamide, 8M urea, 45μM trisborate buffer, pH 7.0 and / or b) HPLC analysis: Waters GenPak FAX column, gradient CH3CN (400ml), H2O (1, 61 ), NaH ₂ PO (3.1 g), NaCl (11.7 g), pH 6.8 (0.1 M NaCl) according to CH3CN

(400ml), H ₂ 0 (1, 61), NaH ₂ P0 ₄ (3.1 g), NaCl (17.53 g), pH 6.8 (1, 5 M an NaCl) and / or c) capillary electrophoresis using a Beckmann eCApT U100P gel capillary column, 65 cm long, 100 mm inner diameter, with the window 15 cm from one end, buffer 140 μM Tris, 360mM

Borate, 7M urea and / or d) electrospray mass spectrometry with negative ions, which confirmed the expected mass values in all cases.

The methods for analyzing oligonucleotides according to a), b), c) and d) are known to the person skilled in the art. These methods are described, for example, in Schweitzer and Engels "Analysis of oligonucleotides" (in "Antisense - from technology to therapy", a laboratory manual and textbook, Schlingensiepen et al. Ed., Biol. Science Vol. 6 (1997) p. 78-103).

The following oligonucleotides were made (see description) and tested: ON1: 3-'C ^* T ^* T ^* AAGGC ^* AGT ^* AC ^* CG ^* CAG ^* C (K3) SEQ ID NO.10

ON2: 3'-A * C ^* C ^* AC ^* TC ^* TAC ^* GT ^* C ^* TGG ^* TA ^* A (K4) SEQ ID NO.11

ON3: 3'-A * A ^* G ^* AG ^* TC ^* AC ^* TC ^* TC ^* C ^* TAGG ^* C (K5) SEQ. ID NO.19

ON4: 3-'C ^* T ^* T ^* AAGGC ^* AGT ^* AC ^* CG ^* CAG ^* C-FITC-5 '(K6) SEQ ID NO.10

ON5: 3'-G * G ^* CAG ^* TAC ^* CG ^* CAGC ^* G SEQ ID NO.22

ON6: 3'-C * T * T ^* AAGG ^* CAG ^* T ^* A SEQ ID NO.13

ON7: 3'-T * A * AGGC ^* AG ^* TA ^* C ^* C SEQIDNO. 14

ON8: 3'-G * G ^* CAG ^* TAC ^* C ^* GC ^* A SEQIDNO. 15

ON9: 3'-C * A * G ^* TAC ^* CG ^* CAG ^* C SEQ ID NO.23

ON10: 3'-A ^* G ^* TAC ^* CG ^* CAG ^* C ^* G SEQIDNO.16

ON11: 3'-C ^* C ^* G ^* CAG ^* CGT ^* CG ^* G SEQIDNO.17

ON12: 3'-G ^* C * AGC ^* GT ^* CGG ^* T ^* T SEQIDNO.18

ON 13: 3'-A ^* A * G ^* AG ^* TC ^* AC ^* TC ^* TC ^* C * TAGG ^* C-Flu-5 '(comparison 1)

SEQIDNO. \ 9 ON14: 3 "-G ^* G ^* C ^* AG ^* TAC ^* CGC ^* AG ^* CGT ^* CG * G SEQ ID NO.12

ON15: 3'-C * TTAAG G ^* CAG ^* T ^* A-FITC SEQ ID NO.13

in which

" ^* " represents a phosphorothioate internucleoside bridge, and FITC represents a fluorescent marker.

ON1 to ON 12 were tested in a cell-based assay for their effectiveness in inhibiting the proliferation of REH leukemia cells. ON1, ON2, ON64-ON71 are antisense oligonucleotides that target the translational start region of eg5 mRNA are targeted. ON4 is the 5'-fluorescein labeled analogue of ON1. ON3 is a reference oligonucleotide.

The results of the proliferation inhibition experiment are shown in Figure 1.

Example 2: Determination of the antiproliferative activity of the eg5 antisense oligonucleotides

The REH cells (human pre-B leukemia cells, DSM ACC 22) and the A549 tumor cells were in OptiMEM (Gibco BRL) with 10% fetal calf serum (FCS, GIBCO-BRL) at 37 ° C under 5% CO ₂ cultured. The cell density for the assay was approximately 1 x 106 / ml. The oligonucleotides (0.17mM) were mixed with Cellfectin (0.83 mg / ml; Gibco-BRL) for complex formation in order to improve the uptake by the cells. The oligonucleotide / cellfectin complex was incubated with the cells in the absence of serum in 24-well plates for 4 hours. The oligonucleotide / cellfectin complex was then removed and serum was added so that the final concentration was 10%. After incubation for 96 hours at 37 ° C. under 5% CO ₂ , the cell density was measured with Casy 1 (from Sch Sharp). For this purpose, the cells in each well were mixed well and immediately diluted 1: 100 with Casyton. The mean values of the cell density were determined from 3 individual wells with the same oligonucleotide concentration. The results of the antiproliferative activity are shown in Figure 1.

Table 1: Nucleotide sequence of human eg5 (SEQ ID NO. 20)

1 GAATTCCGTC ATGGCGTCGC AGCCAAATTC GTCTGCGAAG AAGAAAGAGG

51 AGAAGGGGAA GAACATCCAG GTGGTGGTGA GATGCAGACC ATTTAATTTG 301 GCAGAGCGGA AAGCTAGCGC CCATTCAATA GTAGAATGTG ATCCTGTACG

151 AAAAGAAGTT AGTGTACGAA CTGGAGGATT GGCTGACAAG AGCTCAAGGA

201 AAACATACAC TTTTGATATG GTGTTTGGAG CATCTACTAA ACAGATTGAT

251 GTTTACCGAA GTGTTGTTTG TCCAATTCTG GATGAAGTTA TTATGGGCTA

301 TAATTGCACT ATCTTTGCGT ATGGCCAAAC TGGCACTGGA AAAACTTTTA 351 CAATGGAAGG TGAAAGGTCA CCTAATGAAG AGTATACCTG GGAAGAGGAT 401 CCCTTGGCTG GTATAATTCC ACGTACCCTT CATCAAATTT TTGAGAAACT

451 TACTGATAAT GGTACTGAAT TTTCAGTCAA AGTGTCTCTG TTGGAGATCT 501 ATAATGAAGA GCTTTTTGAT CTTCTTAATC CATCATCTGA TGTTTCTGAG

551 AGACTACAGA TGTTTGATGA TCCCCGTAAC AAGAGAGGAG TGATAATTAA

601 AGGTTTAGAA GAAATTACAG TACACAACAA GGATGAAGTC TATCAAATTT 651 TAGAAAAGGG GGCAGCAAAA AGGACAACTG CAGCTACTCT GATGAATGCA

701 TACTCTAGTC GTTCCCACTC AGTTTTCTCT GTTACAATAC ATATGAAAGA

751 AACTACGATT GATGGAGAAG AGCTTGTTAA AATCGGAAAG TTGAACTTGG

801 TTGATCTTGC AGGAAGTGAA AACATTGGCC GTTCTGGAGC TGTTGATAAG

851 AGAGCTCGGG AAGCTGGAAA TATAAATCAA TCCCTGTTGA CTTTGGGAAG 901 GGTCATTACT GCCCTTGTAG AAAGAACACC TCATGTTCCT TATCGAGAAT

951 CTAAACTAAC TAGAATCCTC CAGGATTCTC TTGGAGGGCG TACAAGAACA

1001 TCTATAATTG CAACAATTTC TCCTGCATCT CTCAATCTTG AGGAAACTCT

1051 GAGTACATTG GAATATGCTC ATAGAGCAAA GAACATATTG AATAAGCCTG

1101 AAGTGAATCA GAAACTCACC AAAAAAGCTC TTATTAAGGA GTATΛCGGAG 1151 GAGATAGAAC GTTTAAAACG AGATCTTGCT GCAGCCCGTG AGAAAAATGG

1201 AGTGTATATT TCTGAAGAAA ATTTTAGAGT CATGAGTGGA AAATTAACTG 1251 TTCAAGAAGA GCAGATTGTA GAATTGATTG AAAAAATTGG TGCTGTTGAG

1301 GAGGAGCTGA ATAGGGTTAC AGAGTTGTTT ATGGATAATA AAAATGAACT

1351 TGACCAGTGT AAATCTGACC TGCAAAATAA AACACAAGAA CTTGAAACCA 1401 CTCAAAAACA TTTGCAAGAA ACTAAATTAC AACTTGTTAA AGAAGAATAT

1451 ATCACATCAG CTTTGGAAAG TACTGAGGAG AAACTTCATG ATGCTGCCAG

1501 CAAGCTGCTT AACACAGTTG AAGAAACTAC AAAAGATGTA TCTGGTCTCC

1551 ATTCCAAACT GGATCGTAAG AAGGCAGTTG ACCAACACAA TGCAGAAGCT

1601 CAGGATATTT TTGGCAAAAA CCTGAATAGT CTGTTTAATA ATATGGAAGA 1651 ATTAATTAAG GATGGCAGCT CAAAGCAAAA GGCCATGCTA GAAGTACATA

1701 AGACCTTATT TGGTAATCTG CTGTCTTCCA GTGTCTCTGC ATTAGATACC

1751 ATTACTACAG TAGCACTTGG ATCTCTCACA TCTATTCCAG AAAATGTGTC

1801 TACTCATGTT TCTCAGATTT TTAATATGAT ACTAAAAGAA CAATCATTAG 1851 CAGCAGAAAG TAAAACTGTA CTACAGGAAT TGATTAATGT ACTCAAGACT 1901 GATCTTCTAA GTTCACTGGA AATGATTTTA TCCCCAACTG TGGTGTCTAT 1951 ACTGAAAATC AATAGTCAAC TAAAGCATAT TTTCAAGACT TCATTGACAG

2001 TGGCCGATAA GATAGAAGAT CAAAAAAAAA GGAACTCAGA TGGCTTTCTC 2051 AGTATACTGT GTAACAATCT ACATGAACTA CAAGAAAATA CCATTTGTTC

2101 CTTGGTTGAG TCACAAAAGC AATGTGGAAA CCTAACTGAA GACCTGAAGA

2151 CAATAAAGCA GACCCATTCC CAGGAACTTT GCAAGTTAAT GAATCTTTGG 2201 ACAGAGAGAT TCTGTGCTTT GGAGGAAAAG TGTGAAAATA TACAGAAACC

2251 ACTTAGTAGT GTCCAGGAAA ATATACAGCA GAAATCTAAG GATATAGTCA

2301 ACAAAATGAC TTTTCACAGT CAAAAATTTT GTGCTGATTC TGATGGCTTC

2351 TCACAGGAAC TCAGAAATTT TAACCAAGAA GGTACAAAAT TGGTTGAAGA

2401 ATCTGTGAAA CACTCTGATA AACTCAATGG CAACCTGGAA AAAATATCTC 2451 AAGAGACTGA ACAGAGATGT GAATCTCTGA ACACAAGAAC AGTTTATTTT

2501 TCTGAACAGT GGGTATCTTC CTTAAATGAA AGGGAACAGG AACTTCACAA

2551 CTTATTGGAG GTTGTAAGCC AATGTTGTGA GGCTTCAAGT TCAGACATCA

2601 CTGAGAAATC AGATGGACGT AAGGCAGCTC ATGAGAAACA GCATAACATT

2651 TTTCTTGATC AGATGACTAT TGATGAAGAT AAATTGATAG CACAAAATCT 2701 AGAACTTAAT GAAACCATAA AAATTGGTTT GACTAAGCTT AATTGCTTTC

2751 TGGAACAGGA TCTGAAACTG GATATCCCAA CAGGTACGAC ACCACAGAGG

2801 AAAAGTTATT TATACCCATC AACACTGGTA AGAACTGAAC CACGTGAACA

2851 TCTCCTTGAT CAGCTGAAAA GGAAACAGCC TGAGCTGTTA ATGATGCTAA

2901 ACTGTTCAGA AAACAACAAA GAAGAGACAA TTCCGGATGT GGATGTAGAA 2951 GAGGCAGTTC TGGGGCAGTA TACTGAAGAA CCTCTAAGTC AAGAGCCATC

3001 TGTAGATGCT GGTGTGGATT GTTCATCAAT TGGCGGGGTT CCATTTTTCC 3051 AGCATAAAAA ATCACATGGA AAAGACAAAG AAAACAGAGG CATTAACACA

3101 CTGGAGAGGT CTAAAGTGGA AGAAACTACA GAGCACTTGG TTACAAAGAG

3151 CAGATTACCT CTGCGAGCCC AGATCAACCT TTAATTCACT TGGGGGTTGG 3201 CAATTTTATT TTTAAAGAAA AACTTAAAAA TAAAACCTGA AACCCCAGAA

3251 CTTGAGCCTT GTGTATAGAT TTTAAAAGAA TATATATATC AGCCGGGCGC

3301 GTGGCTCTAG CTGTAATCCC AGCTAACTTT GGAGGCTGAG GCGGGTGGAT

3351 TGCTTGAGCC CAGGAGTTTG AGACCAGCCT GGCCAACGTG CGCTAAAACC

3401 TTCGTCTCTG TTAAAAATTA GCCGGGCGTG GTGGGCACAC TCCTGTAATC 3451 CCAGCTACTG GGGAGGCTGA GGCACGAGAA TCACTTGAAC CCAGAAGCGG 3501 GGTTGCAGTG AGCCAAAGGT ACACCACTAC ACTCCAGCCT GGGCAACAGA

3551 GCAAGACTCG GTCTCAAAAA TAAAATTTAA AAAAGATATA AGGCAGTACT

3601 GTAAATTCAG TTGAATTTTG ATATCTACCC ATTTTTCTGT CATCCCTATA

3651 GTTCACTTTG TATTAAATTG GGTTTCATTT GGGATTTGCA ATGTAAATAC

3701 GTATTTCTAG TTTTCATATA AAGTAGTTCT TTTAGGAATT C

Table 2: SEQ ID NO. 21: P. falciparum sequence (partial sequence; Genbank, ID Z98551).

TTTTTTTTTTTTTATTCCTTGGATGTTCTTGGTAGTTTAAATTTTTTATTTTTGTAGTTTTC TTCTTTTATACGTTTTAAAGCAGGGGATGCCTTTTTAGGAAATGCCCTATTTTCAATAGCTT TAATTTTTGTAGATTGAAATTTATTATTATTATTATTATTATTGTTGTTGTTGTTGTTGTTG TTGTTGTTGTTATTATTTGAATAATTATTTGTTATATGAACATTTTGAACATTTATATTTCT CTTTCTTTCATATTCTTTTAAACTTGTTACACTCATATTTTCTGTATTTACATCAAATCTTT TATTATGTTGATTGTTATTTAAATAATTTAATTCTTGATATGTTTCATCTATTGGTTGTATA GGATTATCCGTTGTATTCTTATTATATAGCATATATTCATTTAAGGGTAGATTATTGTGATT AGTTTTTACATTTAATTTATTTTTATCACCTTTATTATTTATATTATGAGGTATACTACTAT TCGTTGTATGATCATTTAAACTATTGTAACGAGAGTAATTATTTTCATGCGCTACAATTTTA TCATCTTGAATAAGAAATTGGAAGTTTTCATCGATTTGTTCAAATACTTTACTTAAATCTAT ATCATGTGTTGTTGTAATTTGTTCTATCTCTTTCATCAAGGTATTTTTAACTTCCAAGTATA AATTTTGTCTTATGATATCATCATTATAAAGATAATAATTATGATGATCACCTTGATCTATT TTATTATCATCATTATAAAGATAATAATTATGATGATCACCTTGATCCATTTTATTATCATC ATTATAAAGATAATTATTATGATCATGACCTTGATCCATTTTATTATCATCATTATAAAGAT AATTATTATGATCATGACCTTGATCCATTTTATTATCATCATTATAAAGATAATTATTATGA TCATGACCTTGATCCATTTTATTATCATCATAATTATTATTGTCACCATTTTTAT TATTGTC ATGATCATTTTTATTATTGTCACCATTTTTATTATTATCATGATTATTTTTATTATTATCAT GATTATTTTTATTATTATCATGATTATTTTTATTATTATCATCATTTTTATTATTATCATAA TTCGTGTCGTAAGTCGAATCCCTATTTAGTGATGTGATTTTCATCGGAGTAAACATATCTAT GACATTCACAAACGTTTCCCTTATCCTTTGTACATCATCCTTTATATTTAGATAAAATTCAT CATCCATATTTTCCATGAGATCATAACTTGATGTACTTGGAATGTCTTGTAAGTAATCTTTT TTTTTTAATATATCTATTAATTCTGCTATATACATATTACATTTGTTTAAATTTTGTTCAAA TATATTATTAAAAAGTTTTATATTTTCATTAGACTTTAACATATGTATACGACGTCCCCCTT TTTGTTCTTGTGATTCTTTATTTTTATTTTGTAAAATCTTTTCAGATATAACGTTATATAAC TTTCGTTTCTCTATTTTGTTTATATTAGTTTGACTTGTAAAGTTATTTATGATTTTATCAAT ATTTAGATTATTTGTATATAATAAATTATTATAAATATTTAAAGTATCATTTAAACATTTGC TGTGTTCCTTTTCTTCAATATAACTTTTTCTTTTTAAATAAGATAATATGTTATATAAAACA GTATGATAATTTGTTATCTTCCTTTTAATATCATTATTAATATTATTATATTCCTTTTCATC ATTAATATTGCATTCAGAAAAATGTTGTATAGTATCATCTATCTTTTTTACAGAATTCATAA AAACAGATTTATAATTTTTTTTTGACTTATCATATAATTCTTTATTTAATAAATCGAACTTG TTATTCATTTTTTCATAAATATCTTCCACATTTTTATTTAT _A AGTAATTCAATATCTTTCAA AATATTTTCTTTAAATTCTTGTATATCTTCATTTATCATTTTTTT _A TAATTATTAATTATAA TATTATCCTCCTTTTCAAAAACATCATATTTTTTATAA _A TAT _A TT _CA ATGTTGTCATTCATA ATCTTCTTGTCCTTATCCCAATGTATATATTTTTCATGACATTTTTTTTCTAGTAACATAAA TGATTCGTTTAAAAAAATATGAAATATATTACATATACTTTTAATATATTGTATTAATGATT TTGCATTATATAACTTTTTTTCAGATTCGTGATTATCTAAA TTTGTATAATATCTTCACCT TGTCTATTTAATAATAAATCTTTTATAAATTCTTTATTCTCAGGATAATTAAATGATTCCTC TATATGGTCAAATGGCATCTCATTATTTTCTTCTTTACCATATTGTTTTTGACATGTTTTTC CTTCACCATTTTGTTTTTCACATATTATTCCTTCACCGTTTTGTTTTTCACATATTATCTCG TCACCGTTTTGTTTTTCACATATTATCTTT CACCACTTTGTTTTTCACATATTATCTCTTC ACCGTTTTGTTTTTCACATATTATCTTTTCACCACTTTGTTTTTCACATATTATCTTTTCAC CACTTTGTTTTTCACATATTATCTTTTCACCACTTTGTTTTTCTTTTTTTAATCCGTTTGTA TTATATACACCAATAATTGCTGGCATTTTCTGCTTGGCTTCATCACTTATATGTGGTATGTT TATTTTACATTGTGATATTTCCTTTTTAATATTTTCGGAGAGAGAAAAGTAATCATGATCAT ATTTTTGTAAAATATCCATATGGTCCAGTATAAAATTCAGAGTATCATTATATTTAAAATTA ATGTTACTATTGAGTTCTTCAAAATGGTTAATATAATCATTGTATGATTTTTTATTTTGTAC TAGATAATTTTTGGTATCATCTAAAATGAATAAGATGGTTTTACATATATCGTTTAAAAGAT G _A TTTTCTTGATGAATATTTTTTTTTATATTTAATAGATTATCATGCATTATATTAGACATA TTTGTTTTAATTTGTTGAAAAGATTTTTTTTGATTTATAAAATTTTCTTCTAAAGAATGATA TTTATTTAATAAGAATTGTGTAATATATTTTTCTTCTATTATTTTTTTAATTA ATATTTGAT GAAATGCTTGTATATTTTTATATTTTTGAATAGTATCTTTTAAAAAGAAAAATATTTTATTT TGTAGATCATCTGTATTATCCATTTTATTTAATAAATTTTTTATTTTTTTACTTTTTTCAAA TAAAATTTTTTCTTTTTCTAATAATATTTCTTTATTTTTCTTTAGACTATTTTGTATATTAT TATATTCTTCTGTATCAAGATAAACACCTCTCTTTTCTCTGCTTAAATTCAGTGCATTTCTT AACTTTTCGATTTCATTATTTAAATCCTTTATTTTTAATTGTTTCGTTGTTTTTATATTTAT CTCGGGTCTATTCTTAATATTCTTAGCTCGAAAGACATAATCTAAAGTGCTTAAAGTCTCAT CAATACATAAAGAGGAGGGTGATATAGTGGCGACAATAAAAGTCTTCGTTTTCCCACCTAAC GAATCTTGTAATAATCTGGTTAATTTAGAATCTCTGTAAGGAATATAAGATGAATTCTCAAT CAACGAATTAATAACTCTACCTAAGGTTAATAAAGATTGATTTATATTACAACTTTCTTGTT GTCTAATTTTTAAAGAACCATAAGAGCTTTTCAAAGCATTTTCACTACCCGCTAAATCAACT AAATTTAATTTTCCTATTTTTGTTATACTTTCTCCTACATTATTTATATCTTTTATAATTAA TGTTATAGTAAAAATCGAATGACTTCTACTCGATTTTTTATTATAAGCCGTTTCAGCTGTCC TTCTTTTTTTAATAGCTGAACATATAATATAATATATTTCTTCAAAAGAATTAATACTTTTT TCTTCTAACTTATCAACATTTAATCCTTTACTTTTATTATTACTATCTTCATATATTCGAAG TTTCATATTTTCATTTGTTGAACTTAATAAATCACATAATTCTTCATTATATATTTCTAGAT AGCTAATTTTTATATTAAAATCGTACATATTCTTATCATCAAATG TTTGATACATATCATTA TTCCTATTTTTATCTACACTACATTTTTGTACAACATCACAAGTAATATCTCTACTCTTTTC GTTA _A CTAACAAATTGTTAGGTTCTTTATTAATTTTTAAATTATT _A TAAATATCATTTTTAT CA _A T _A TTTATTTTGTTACATAATAAATTATTATAATTATTATTAAT _A ATATTATTATTGTTA CC _A TT _A GTTTCCTTATTTATTACATTTATATGTTCGTTATCCTTTTCATCAAATATATTCTT TTTTCCTTTAAAATGTCGAATCTTTTCTTCTTTCCTTTTATTT _AA TATATCGAATATTCTTT TCGTAACTCGAAATATAAGTCCAGTATCCTCATTCTCACAAAGTTCATAGCAATAGCTGATG TCGCTATTAATACTTTCATTCAAATCCACCTTTTTATTATTATCATATTGTTTCAGGTGTTC

TAGTATTTTCCCTTCCATAGTATAGGTCTTACCCGTCCCGGTCTGTCCATAGCAGAACAGCG

TACAATTGAATCCTTGCAAAACCTGAAGCGGCGAACAAAAAAAAAAAAAAAAAAAAATATAT

ATATATATGTACATGTATATTTATATGTATATGTATATATATATGTATAGTTATATGTATTT

TTATTTTTATTTTTATTTTTATATTTATTTTTATTTTTATATTTATTTTTATTTTTATATTT _{ATTTTTATTTTTATATTTATATTTATATA}

CCATATATTTTTTTTTTTTAATCTATTTAATAAAACATTATTATGATATACGCAGAGGTGAT

ATATACATGGTATTTATTTATTTTTTTTTATATATTTTTCAT

TTGTTTCGTAGGAATATTCTTTTTTTTTCTGCACATATATTTCACTATCCATATAATATCAT

AATACATCATGGAATAATTTATATATATATATATATATATGTATATTTTATTTTTACCTCAT

CTACTATTTGGTAAATATAATTATTGAACAAAGTTTTCTGATCCACATCTTTATCACATGCA

TAATCAAAACTATATTTTTTTTCGTATATTTCATTGTTTCTATTAATTGTTAATATAACCTC

ATTATTATTAATTCGAACTACCTCTTCATTATTTATATCGTTTTTTTCTTTTTCATTTAATG

GTCTACACCTTACGATAACTTTTATATTTACGCAACTTGATTTATCATTATTATAAGAATTT

CTGAGCATTTTACTTTTATTCAAATAAT

Table 3:

Sequence homology: Comparison of the human eg5 sequence with the Plasmodium falciparum eg5 sequence

1 60 human. SEQ GAATTCCGTCAT GGCGTC .... GCAGCC .AAATTC ... GTCTGCGAAGAAG P ASMO. SEQ TTTTTTTTTTTTTATTCCTTGGATGTTCTTGGTAGTTTAAATTTTTTATTTTTGTAGTTT

61 120 human. SEQ AAAGA .... GGAGAAGGGGAAGAACATCCAGGTGGTGGTGAGATGCAGACC PLASMO. SEQ TCTTCTTTTATACGTTTTAAAGCAGGGGATGCCTTTTTAGGAAATGCCCTATTTTCAATA

121 180 human. SEQ A. TTTAATTTGGCAGAGCGGAAAGCTAGCGCCCAT. TCAATAGTAGAATGTGATCCTGTA

PLASMO. SEQ GCTTTAATTTTTGTAGATTGAAATTTATTATTATTATTATTATTATTGT. TGTTGTTGTT

181 240 human .SEQ CGAAAAGAAGTTAGTGT .ACGAACTGGAGGATTGGCTG .. ACAAGAGCTCAAGGAAAACA PLASMO. SEQ GTTGTTGTTGTTGTTGTTATTATTTGAATAATTATTTGTTATATGAACATTTTGAACATT

241 300 human. SEQ TACACTTTTGAT ATGGTGTTTGGAGC ATCTACTAAAC .. AG

PLASMO. SEQ TATATTTCTCTTTCTTTCATATTCTTTTAAACTTGTTACACTCATATTTTCTGTATTTAC

301 360 human. SEQ ATTGA .. TGTTTACCG .... AAGTGTTGTTTG TCCAATTCTGGATGAAGT. AT.

PLASMO. SEQ ATCAAATCTTTTATTATGTTGATTGTTATTTAAATAATTTAATTCTTGATATGTTTCATC 361 420 human. SEQ TATGGGCTATA .... ATTGCAC .... TATCTTTGC. GTATGGC. CAAACT GG

PLASMO. SEQ TATTGGTTGTATAGGATTATCCGTTGTATTCTTATTATATAGCATATATTCATTTAAGGG

421 480 human. SEQ APPROX. CTG. GAAAAACTTTTACAATGGA ... AGGTGAAAGGTC ACCTA ....

PLASMO. SEQ TAGATTATTGTGATTAGTTTTTACATTTAATTTATTTTTATCACCTTTATTATTTATATT

Illustration 1 :

This figure summarizes the results of Examples 1 + 2:

The effect of oligonucleotides ON1 to ON12 (eg5 antisense) is shown

Inhibition of REH cell proliferation (in percent).

■ 1st experiment ♦ 2nd experiment CF: cellfectin comparison

Claims

Expectations:

1. Antisense oligonucleotide or one of its derivatives, characterized in that it corresponds to part of an eg5 coding sequence.

2. antisense oligonucleotide or one of its derivatives according to claim 1, characterized in that it corresponds to 8-100 nucleotides of an eg5 coding sequence.

3. antisense oligonucleotide or one of its derivatives according to one of claims 1 or 2, characterized in that it has a length of 8 to 20 nucleotides.

4. Antisense oligonucleotide or one of its derivatives according to one or more of claims 1 to 3, characterized in that it corresponds to a part of a human coding sequence eg5 and / or a Plasmodium falciparum eg5.

5. antisense oligonucleotide or one of its derivatives according to one or more of claims 1 to 4, characterized in that it is one of the sequences SEQ ID NO. 1, SEQ ID NO. 2, SEQ ID NO. 3, SEQ ID NO. 4, SEQ ID NO. 5, SEQ ID NO. 6, SEQ ID NO. 7, SEQ ID NO. 8 or SEQ ID NO. 9 has

SEQ ID NO. 1 ^' 5'-CGACGCCATGACGGAATTC-3'

SEQ ID NO. 2 5'-AATGGTCTGCATCTCACCA-3 '

SEQ ID NO. 3 5'-GGCTGCGACGCCATGACGG-3 '

SEQ ID NO. 4 5'-ATGACGGAATTC-3 'SEQ ID NO. 5 5'-CCATGACGGAAT-3 '

SEQ ID NO. 6 5'-ACGCCATGACGG-3 '

SEQ ID NO. 7 5'-GCGACGCCATGA-3 '

SEQ ID NO. 8 5'-GGCTGCGACGCC-3 'and SEQ ID NO. 9 5'-TTGGCTGCGACG-3 '.

6. antisense oligonucleotide or one of its derivatives according to one or more of claims 1 to 5, characterized in that the oligonucleotide has one or more modifications, each being

Modification, compared to an oligonucleotide made of natural DNA with the same sequence, is located on a specific phosphodiester internucleoside bridge and / or on a specific β-D-2 ^' deoxyribose unit and / or on a specific natural nucleoside base position.

7. antisense oligonucleotide or one of its derivatives according to one or more of claims 1 to 6, characterized in that 1 to 5 terminal nucleotide units at the 5 'end and / or at the 3' end of the oligonucleotide by modified internucleoside bridges which are at 5 ' - and / or are located at the 3 'end of the corresponding nucleoside or is protected.

8. antisense oligonucleotide or one of its derivatives according to one or more of claims 1 to 7, characterized in that at least one internal pyrimidine nucleoside and / or an internucleoside bridge located at the 5 'end and / or at the 3' end of this pyrimidine nucleoside is modified ,

9. antisense oligonucleotide or one of its derivatives according to one or more of claims 1 to 8, characterized in that each modification is independently selected from a) the exchange of the phosphodiester bridge at the 3 ^' and / or 5 ^' end of a nucleoside a modified internucleoside bridge, b) the replacement of the phosphodiester bridge at the 3 ^' and / or 5 ^' end of a nucleoside by a dephospho bridge, c) the exchange of a sugar phosphate residue from the sugar phosphate skeleton by another residue, d) the replacement of a β-D-2 ^' deoxyribose unit by a modified sugar unit, e) the replacement of a natural nucleoside base by a modified nucleoside base, f) the connection to a molecule which influences the properties of the oligonucleotide, g) the connection to a 2'5'-linked oligoadenylate molecule or a derivative thereof, optionally via a suitable linker molecule, and h) the introduction of a 3 ^' -3 ^' and / or a 5 ^' -5 ^' inversion on 3 ^' and / or on 5 ^' end of the oligonucleotide.

10. A method for producing an antisense oligonucleotide or one of its derivatives according to one or more of claims 1 to 9, wherein protected monomers are condensed in a suitable manner on a solid phase.

11. Use of an antisense oligonucleotide or one of its derivatives according to one or more of claims 1 to 9 for inhibiting the expression of eg5.

12. A method for inhibiting the expression of eg5, characterized in that an antisense oligonucleotide or one of its derivatives according to one or more of claims 1 to 9 is brought into contact with and binds to an eg5-coding nucleic acid.

13. A process for the preparation of a pharmaceutical preparation, characterized in that one or more antisense oligonucleotides or one of its derivatives according to one or more of claims 1 to 9 are mixed with a physiologically acceptable carrier and optionally additional substances.

4. Pharmaceutical preparation, characterized in that it contains at least one antisense oligonucleotide or one of its derivatives according to one or more of claims 1 to 9.