Oligonucleotides for the inhibition of human eg5 expression

• US 6,472,521 B1
• Filed: 07/27/2000
• Issued: 10/29/2002
• Est. Priority Date: 07/28/1999
• Status: Expired due to Fees

• Current Assignee: Sanofi-Aventis Deutschland GmbH (Sanofi )
• Original Assignee: Aventis Pharma Deutschland GmbH (Sanofi )
• Inventors: Greiner, Beate, Unger, Eberhard, Uhlmann, Eugen, Gothe, Gislinde, Schwerdel, Marc
• Primary Examiner(s): McGarry, Sean
• Application Number:
  US09/627,122
• Time in Patent Office: 824 Days
• Field of Search: 436/6, 436/375, 436/377, 436/455, 536/23.1, 536/24.1, 536/24.5, 536/25.3, 514/44
• US Class Current: 536/24.5
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 35/02 specific for leukemia
• CPC Class Codes: A61P 35/04 specific for metastasis
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: C07B 2200/11 Compounds covalently bound to a solid support
• CPC Class Codes: C07H 21/00 Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids
• CPC Class Codes: C12N 15/113 Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing when used in plants C12N15/8218
• CPC Class Codes: C12N 2310/315 Phosphorothioates
• CPC Class Codes: C12N 2310/317 with an inverted bond, e.g. a cap structure
• CPC Class Codes: C12N 2310/318 where the PO2 is completely replaced, e.g. MMI or formacetal
• CPC Class Codes: C12N 2310/32 of the sugar
• CPC Class Codes: C12N 2310/33 of the base
• CPC Class Codes: C12N 2310/346 having a combination of backbone and sugar modifications
• CPC Class Codes: Y02A 50/30 Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Abstract

The present invention relates to an oligonucleotide or a derivative thereof which has a sequence that corresponds to a particular fragment of a nucleic acid sequence which encodes human eg5 or a mutant form thereof; the invention further relates to a method of making the oligonucleotide and the use thereof.

Claims

1. An oligonucleotide or a derivative thereof, comprising up to 100 nucleotides, wherein the sequence of the oligonucleotide or derivative thereof comprises at least one of 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.

2. The oligonucleotide or derivative thereof as claimed in claim 1, wherein the oligonucleotide sequence is SEQ ID NO:

1, SEQ ID NO;

2, SEQ ID NO;

3, SEQ ID NO;

4, SEQ ID NO;

5, SEO ID NO;

6, SEQ ID NO;

7. SEQ ID NO;

8, or SEQ ID NO;

9.

3. The oligonucleotide or derivative thereof as claimed in claim 1, wherein the oligonucleotide or derivative thereof has one or more modifications.

4. The oligonucleotide or derivative thereof as claimed in claim 3, wherein said one or more modifications are located at one or more phosphodiester internucleoside bridges, and/or at one or more β

D-2′

-deoxyribose units, and/or at one or more nucleoside bases.

5. The oligonucleotide or derivative thereof as claimed in claim 3, wherein from 1 to 5 terminal nucleotides at the 5′

-end and/or at the 3′

-end of the oligonucleotide or derivative thereof have modified internucleoside bridges located at the 5′

-end and/or the 3′

-end of the nucleotide.

6. The oligonucleotide or derivative thereof as claimed in claim 3, wherein at least one internal pyrimidine nucleoside and/or an internucleoside bridge located at the 5′

-end and/or the 3′

-end of said at least one internal pyrimidine nucleoside is modified.

7. The oligonucleotide or derivative thereof as claimed in claim 3, wherein each modification is independently selected from:

(a) replacement of a phosphodiester bridge at a 3′

- and/or a 5′

-end of a nucleoside by a modified internucleoside bridge;

(b) replacement of a phosphodiester bridge at a 3′

- and/or a 5′

-end of a nucleoside by a dephospho bridge;

(c) replacement of a sugar phosphate residue from a sugar phosphate backbone by another residue;

(d) replacement of a β

D-2′

-deoxyribose unit by a modified sugar unit;

(e) replacement of a natural nucleoside base by a modified nucleoside base;

(f) conjugation to a molecule which modifies one or more properties of the oligonucleotide or derivative thereof selected from ability to penetrate a cell membrane, ability to enter a cell, stability toward nucleases, affinity for an eg5 encoding target sequence, pharmakokinetics, ability to cleave the eg5 encoding target sequence, and ability to crosslink;

(g) conjugation to a 2′

-5′

-linked oligoadenylate molecule or a derivative thereof, optionally via an appropriate linker molecule; and

(h) introduction of a 3′

-3′ and

/or a 5′

-5′

inversion at a 3′

- and/or a 5′

-end of the oligonucleotide or derivative thereof.

8. A method of making the oligonucteotide or derivative thereof as claimed in claim 1, comprising the step of condensing suitably protected monomers on a solid phase.

9. A method of inhibiting eg5 gene expression, comprising the step of contacting the oligonucleotide or derivative thereof as claimed in claim 1 with a nucleic acid sequence encoding an eg5 protein, wherein said oligonucleoude or derivative thereof binds with said nucleic acid sequence.

10. A pharmaceutical composition comprising at least one oligonucleotide or derivative thereof as claimed in claims 1 or 3.

11. A method of making a pharmaceutical composition comprising mixing one or more oligonucleotides or derivatives thereof as claimed in claim 1 with a physiologically acceptable excipient.

12. The method of making a pharmaceutical composition as claimed in claim 11, wherein the composition further comprises an auxiliary substance or additive.