Heparanase specific molecular probes and their use in research and medical applications

• US 20040229834A1
• Filed: 05/24/2004
• Published: 11/18/2004
• Est. Priority Date: 09/02/1997
• Status: Active Grant

• Current Assignee: Iris Pecker, Israel Vlodavsky, Tuvia Peretz, Yael Friedmann
• Original Assignee: Iris Pecker, Israel Vlodavsky, Tuvia Peretz, Yael Friedmann
• Inventors: Pecker, Iris, Friedmann, Yael, Peretz, Tuvia, Vlodavsky, Israel
• Granted Patent: US 7,790,376 B2
• Time in Patent Office: Days
• US Class Current: 514/44
• 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: A61M 15/009 using medicine packages with incorporated spraying means, e.g. aerosol cans pocket atomiser of the injector type A61M11/08
• CPC Class Codes: A61P 13/12 of the kidneys
• CPC Class Codes: A61P 29/00 Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
• CPC Class Codes: A61P 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 37/00 Drugs for immunological or allergic disorders
• CPC Class Codes: A61P 37/02 Immunomodulators
• CPC Class Codes: C07K 16/40 against enzymes
• CPC Class Codes: C12N 9/2402 hydrolysing O- and S- glycosyl compounds (3.2.1)
• CPC Class Codes: C12Q 1/6886 for cancer immunoassay for cancer G01N33/574
• CPC Class Codes: C12Q 2600/112 Disease subtyping, staging or classification
• CPC Class Codes: C12Q 2600/136 Screening for pharmacological compounds
• CPC Class Codes: C12Y 302/01166 Heparanase (3.2.1.166)
• CPC Class Codes: G01N 2333/988 Lyases (4.), e.g. aldolases, heparinase, enolases, fumarase
• CPC Class Codes: G01N 2400/40 Glycosaminoglycans, i.e. GAG or mucopolysaccharides, e.g. chondroitin sulfate, dermatan sulfate, hyaluronic acid, heparin, heparan sulfate, and related sulfated polysaccharides
• CPC Class Codes: G01N 33/573 for enzymes or isoenzymes
• CPC Class Codes: Y10S 977/906 Drug delivery

Abstract

Heparanase specific molecular probes which can be used for detection of the presence, absence or level of heparanase expression and methods of utilizing same are provided.

Claims

1. An oligonucleotide comprising a nucleic acid sequence specifically hybridizable with a polynucleotide sequence encoding a heparanase.

2. The oligonucleotide of claim 1, wherein said oligonucleotide sequence is at least 10 and no more than 650 nucleotides in length.

3. The oligonucleotide of claim 1, wherein said polynucleotide sequence encoding heparanase is as set forth by SEQ ID NO:

1.

4. The oligonucleotide of claim 1, wherein said polynucleotide sequence encoding heparanase is encompassed by nucleotides 728-1351 of SEQ ID NO:

1.

5. The oligonucleotide of claim 1, wherein said polynucleotide sequence is a messenger RNA of said heparanase.

6. The oligonucleotide of claim 1, wherein said oligonucleotide is labeled with a detectable moiety.

7. The oligonucleotide of claim 6, wherein said detectable moiety is selected from the group consisting of a chromogenic moiety, a fluorogenic moiety, a light-emitting moiety and a radioactive moiety.

8. The oligonucleotide of claim 1, wherein the oligonucleotide is selected from the group consisting of a synthetic oligonucleotide, an antisense heparanase RNA and heparanase DNA.

9. The oligonucleotide of claim 1, wherein said nucleic acid sequence includes natural nucleotides and/or nucleotide analogs.

10. A kit for detecting a polynucleotide sequence encoding heparanase, the kit comprising the oligonucleotide of claim 1 and reagents for detecting a hybridization between said oligonucleotide and the polynucleotide sequence encoding heparanase.

11. The kit of claim 10, wherein said oligonucleotide is labeled with a detectable moiety.

12. The kit of claim 10, wherein said detectable moiety is selected from the group consisting of a chromogenic moiety, a fluorogenic moiety, a light-emitting moiety and a radioactive moiety.

13. A method of detecting presence or absence or level of a polynucleotide sequence encoding heparanase in a biological sample, the method comprising:

(a) exposing the biological sample to an oligonucleotide, having a nucleic acid sequence specifically hybridizable with the polynucleotide sequence encoding the heparanase; and

(b) detecting specific hybridization, thereby detecting presence, absence or level of the polynucleotide sequence encoding heparanase in the biological sample.

14. The method of claim 13, wherein said biological sample is a cell, a tissue sample or body fluid.

15. The method of claim 13, wherein said biological sample includes malignant cells.

16. The method of claim 13, wherein said biological sample is derived from solid tumor or a hematopoietic tumor.

17. The method of claim 16, wherein said solid tumor is selected from the group consisting of carcinoma, adenocarcinoma, squameous cell carcinoma, teratocarcinoma, mesothelioma, melanoma, renal cell carcinoma, myeloma, and rabdomyosarcoma and further wherein said hematopoietic tumor is selected from the group consisting of lymphoma and leukemia.

18. The method of claim 16, wherein said solid tumor is a primary tumor, or a metastasis thereof, and is originated from an organ selected from the group consisting of liver, prostate, bladder, breast, ovary, cervix, colon, skin, intestine, stomach, uterus, pancreas.

19. The method of claim 13, wherein said oligonucleotide is selected from the group consisting of a synthetic oligonucleotide, an antisense to heparanase RNA and heparanase DNA.

20. The method of claim 13, wherein said oligonucleotide is labeled with a detectable moiety.

21. The method of claim 20, wherein said detectable moiety is selected from the group consisting of a chromogenic moiety, a fluorogenic moiety, a light-emitting moiety and a radioactive moiety 22. The method of claim 13, wherein the polynucleotide sequence encoding the heparanase is mRNA and said oligonucleotide is designed for directing reverse transcription of said mRNA into cDNA and whereas step (b) is effected by polymerase chain reaction amplification of said cDNA.