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Patent Searching and Data


Title:
DRUG TARGETS IN $i(CANDIDA ALBICANS)
Document Type and Number:
WIPO Patent Application WO/2000/034481
Kind Code:
A2
Abstract:
The present invention is concerned with the identification of genes or functional fragments thereof from $i(Candida albicans) which are critical for growth and cell division and which genes may be used as selective drug targets to treat $i(Candida albicans) associated infections. Novel nucleic acid sequences from $i(Candida albicans) are also provided and which encode the polypeptides which are critical for growth of $i(Candida albicans). Methods for the identification of anti-fungal compounds which inhibit fungal or yeast growth are also contemplated.

Inventors:
CONTRERAS ROLAND HENRI (BE)
NELISSEN BART (BE)
DE BACKER MARIANNE DENISE (BE)
LUYTEN WALTER HERMAN MARIA LOU (BE)
VIAENE JASMINE ELZA (BE)
LOGGHE MARC GEORGE (BE)
VIALARD JORGE EDUARDO (BE)
Application Number:
PCT/EP1999/009833
Publication Date:
June 15, 2000
Filing Date:
December 06, 1999
Export Citation:
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Assignee:
JANSSEN PHARMACEUTICA NV (BE)
CONTRERAS ROLAND HENRI (BE)
NELISSEN BART (BE)
BACKER MARIANNE DENISE DE (BE)
LUYTEN WALTER HERMAN MARIA LOU (BE)
VIAENE JASMINE ELZA (BE)
LOGGHE MARC GEORGE (BE)
VIALARD JORGE EDUARDO (BE)
International Classes:
G01N33/50; A61K31/7088; A61K38/00; A61K38/16; A61K45/00; A61P15/02; A61P31/10; C07K14/40; C07K16/14; C12N1/15; C12N1/19; C12N1/21; C12N5/10; C12N15/09; C12N15/31; C12Q1/02; C12Q1/68; G01N33/15; C12R1/725; (IPC1-7): C12N15/31; C07K14/40; G01N33/50; C12Q1/68; A61K31/7088; A61K38/16; C07K16/14
Domestic Patent References:
WO1997036925A11997-10-09
WO1997037230A11997-10-09
WO1996036707A11996-11-21
Other References:
DATABASE EMBL - EMGSS12 [Online] Entry/Acc.no. B89355, 19 March 1998 (1998-03-19) ADAMS, M. D. ET AL.: "RPCI11-27L12.TPB RPCI-11 Homo sapiens genomic clone RPCI-11-27L12, genomic survey sequence." XP002139078
DALY S ET AL: "Isolation and characterization of a gene encoding alpha-tubulin from Candida albicans" GENE: AN INTERNATIONAL JOURNAL ON GENES AND GENOMES,GB,ELSEVIER SCIENCE PUBLISHERS, BARKING, vol. 187, no. 2, 18 March 1997 (1997-03-18), pages 151-158, XP004093273 ISSN: 0378-1119
Attorney, Agent or Firm:
Baldock, Sharon Claire (Verulam Gardens 70 Gray's Inn Road, London WC1X 8BT, GB)
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Claims:
Claims
1. A nucleic acid molecule encoding a polypeptide which is critical for survival and growth of the yeast Candida albicans and which nucleic acid molecule comprises any of the sequences of nucleotides illustrated in Seq ID Nos 1 to 9.
2. A nucleic acid molecule encoding a polypeptide which is critical for survival and growth of the yeast Candida albicans and which nucleic acid molecule comprises any of the sequences of nucleotides illustrated in Seq ID Nos 1 to 3.
3. A nucleic acid molecule encoding a polypeptide which is critical for survival and growth of the yeast Candida albicans and which nucleic acid molecule comprises any of the sequences of nucleotides illustrated in Seq ID Nos 1 or 2 and fragments or derivatives of said nucleic acid molecules.
4. A nucleic acid molecule according to any of claims 1 to 3 which is mRNA.
5. A nucleic acid molecule according to any of claims 1 to 3 which is DNA.
6. A nucleic acid molecule according to claim 5 which is cDNA.
7. A nucleic acid molecule capable of hybridising to the molecules according to any of claims 1 to 6 or the sequences illustrated in any of Seq ID Nos 1 to 9 under high stringency conditions.
8. An antisense molecule comprising a nucleic acid molecule capable of hybridising to the molecules according to any of claims 1 to 6 or the sequences illustrated in any of Seq ID Nos 1 to 9.
9. Cells containing a nucleic acid molecule according to any of claims 1 to 8, wherein said cells are bacterial or eukaryotic.
10. A polypeptide encoded by the nucleic acid molecule according to any of claims 1 to 7 or the sequences illustrated in any of Seq ID Nos 1 to 9.
11. A polypetide having any of amino acid sequences illustrated in any of Seq ID Nos 14 to 20.
12. R recombinant DNA construct comprising a nucleic acid molecule according to claim 5 or 6.
13. A recombinant DNA construct comprising a nucleic acid molecule according to claim 5 or 6 wherein said nucleic acid molecule is inserted in the antisenseorientation.
14. P recombinant DNA construct according to claim 12 or 13 wherein said recombinant DNA construct is an expression vector.
15. A construct according to claim 14 which comprises an inducible promoter.
16. A construct according to claim 14 or 15 which comprises a sequence encoding a reporter molecule.
17. Cells containing a recombinant DNA construct according to any of claims 12 to 16, wherein said cells are bacterial or eukaryotic.
18. A nucleic acid molecule according to any of claims 1 to 8 or the nucleotide sequences illustrated in Seq ID Nos 1 to 9 for use as a medicament.
19. Use of a nucleic acid molecule according to any of claims 1 to 8 or the sequences illustrated in Seq ID Nos 1 to 9 in the preparation of a medicament for treating Candida albicans associated diseases.
20. A polypeptide according to claim 10 or 11 for use as a medicament.
21. Use of a polypeptide according to claim 10 or 11 in the preparation of a medicament for treating Candida albicans associated infections.
22. A pharmaceutical composition comprising a nucleic acid molecule according to any of claims 1 to 8 or a polypeptide according to claim 10 or 11 together with a pharmaceutically acceptable carrier diluent or excipient therefor.
23. A Candida albicans cell comprising an induced mutation in the DNA sequence encoding the polypeptide according to claim 10.
24. A method of identifying compounds which selectively modulate expression or functionality of polypeptides or metabolic pathways in which these polypeptides are involved and which are crucial for growth and survival of Candida albicans, which method comprises: (a) contacting a compound to be tested with one or more Candida albicans cells having a mutation in a nucleic acid molecule according to any of claims 1 to 8 which mutation results in overexpression or underexpression of said polypeptides in addition to contacting one or more wild type Candida albicans cells with said compound, (b) monitoring the growth and/or activity of said mutated cell compared to said wild type; wherein differential growth or activity of said one or more mutated Candida cells is indicative of selective action of said compound on a polypeptide or another polypeptide in the same or a parallel pathway.
25. A compound identifiable according to the method of claim 24.
26. A compound according to claim 25 for use as a medicament.
27. Use of a compound according to claim 25 in the preparation of a medicament for treating Candida albicans associated diseases.
28. A pharmaceutical composition comprising a compound according to claim 25 together with a pharmaceutically acceptable carrier, diluent or excipient therefor.
29. A method of identifying DNA sequences from a cell or organism which DNA encodes polypeptides which are critical for growth or survival of said cell or organism, which method comprises: (a) preparing a cDNA or genomic library from said cell or organism in a suitable expression vector which vector is such that it can either integrate into the genome in said cell or that it permits transcription of antisense RNA from the nucleotide sequences in said cDNA or genomic library. (b) selecting transformants exhibiting impaired growth and determining the nucleotide sequence of the cDNA or genomic sequence from the library included in the vector from said transformant.
30. A method according to claim 29 wherein said cell or organism is a yeast or filamentous fungus.
31. A method according to claim 29 or 30 wherein said cell or organism is any of Saccharomyces cerevisiae, Schizosaccharomyces pombe or Candida albicans.
32. An antibody capable of binding to a polypeptide according to claim 10 or 11.
33. An oligonucleotide comprising a fragment of from 10 to 120 contiguous nucleotides of a nucleic acid molecule according to any of claims 1 to 8.
34. An oligonucleotide according to claim 33 comprising a fragment of from 10 to 50 contiguous nucleotides.
Description:
INTERNATIONAL SEARCH REPORT interrnal Application No PCT/EP 99/09833 C. (Continuation) DOCUMENTS CONSIDERED TO BE RELEVANT Category Citation of document, with indication, where appropriate, of the relevant passages Relevant to claim No. A WO 96 36707 A (UNIV ROMA ; IST SUPERIORE SANITA (IT) ; CASSONE ANTONIO (IT) ; VALLE) 21 November 1996 (1996-11-21) the whole document A DALY S ET AL :"Isolation and characterization of a gene encoding alpha-tubulin from Candida albicans" GENE : AN INTERNATIONAL JOURNAL ON GENES AND GNOMES, GB, ELSEVIER SCIENCE PUBLISHERS, BARKING, vol. 187, no. 2, 18 March 1997 (1997-03-18), pages 151-158, XP004093273 ISSN : 0378-1119 the whole document 1 Ir lational applioation No. INTERNATIONAL SEARCH REPORT PCT/EP 99/09833 Box) Observations where certain claims were found unsearchable (Continuation of item 1 of first sheet) This International Search Report has not been established in respect of certain claims under Article 17 (2) (a) for the following reasons : 1. Claims Nos. : because they relate to subject matter not required to be searched by this Authority, namely : 2. Claims Nos. : 25-28 because they relate to parts of the international Application that do not comply with the prescribed requirements to such an extent that no meaningful international Search can be carried out, specifically : see FURTHER INFORMATION sheet PCT/ISA/210 3. Claims Nos. : because they are dependent claims and are not drafted in accordance with the second and third sentences of Rule 6. 4 (a). Box 11 Observations where unity of invention is lacking (Continuation of item 2 of first sheet) This International Searching Authority found multiple inventions in this international application, as follows : 1. ! ! As ait required additional search fees were timely paid by the applicant, this international Search Report covers all searchable claims. 2. As all searchable claims could be searched without effort justifying an additional fee, this Authority did not invite payment of any additional fee. 3. As only some of the required additional search fees were timely paid by the applicant, this International Search Report covers only those claims for which fees were paid, specifically claims Nos. : 4. No required additional search fees were timely paid by the applicant. Consequently, this Intemational Search Report is restricted to the invention first mentioned in the claims ; it is covered by claims Nos. : 1-28 and 32-34, all partially Remark on Protest The additional search fees were accompanied by the applicant's protest. Cl No protest accompanied the payment of additional search fees. u FURTHER INFORMATION CONTINUED FROM PCT/ISA/210 Continuation of Box 1. 2 Claims Nos. : 25-28 Present claims 25-28 relate to a compound defined by reference to a desirable characteristic or property, namely its identifyability by the method of claim 24.

The application does not provide support within the meaning of Article 6 PCT and/or disclosure within the meaning of Article 5 PCT for any such compounds. In the present case, the claims so lack support, and the application so lacks disclosure, that a meaningful search for these claims is impossible.

The applicant's attention is drawn to the fact that claims, or parts of claims, relating to inventions in respect of which no international search report has been established need not be the subject of an international preliminary examination (Rule 66. 1 (e) PCT). The applicant is advised that the EPO policy when acting as an International Preliminary Examining Authority is normally not to carry out a preliminary examination on matter which has not been searched. This is the case irrespective of whether or not the claims are amended following receipt of the search report or during any Chapter II procedure.

FURTHER INFORMATION CONTINUED FROM PCT/ISA/210 1. Claims : Invention 1 : claims 1-28 and 32-34, all partially Nucleic acid molecule comprising seq. ID. 1 or capable of hybridizing thereto and antisense thereof, cell containing said nucleic acid, polypeptide of seq. ID. 10 encoded by said nucleic acid, expression vector comprising said nucleic acid, antibody against said peptide, use of said nucleic acid or protein for preparation of medicament or pharmaceutical composition, C. albicans cell comprising an induced mutation in said DNA sequence, oligonucleotides comprising 10-120 nt of said nucleic acid sequence, and method for identifying compounds which modulate expression of said nucleic acid.

2. Claims : Inventions 2-9 : claims 1-28 and 32-34, all partially and as applicable As invention 1, but limited to the respective nucleic acid sequences 2-9, and polypeptide sequences corresponding thereto in as far as they are provided (see table 1 of the description).

3. Claims : Invention 10 : claim 29-31 Method for identifying DNA sequences from a cell or organism, which encode polypeptides which are critical for growth and survival for said cell or organism, comprising screening a library of nucleic acids using a vector that either integrates into the genome of said cell or organism, or that permits expression of antisense RNA, and selecting growth- impaired cells or organisms. INTERNATIONAL SEARCH REPORT !----------------- ! Intema'nal Appllcation No li nation on patent tamily members PCT/EP 99/09833 Patent document Publication Patent family Publication cited in search report date member (s) date WO 9736925 A 09-10-1997 CA 2250129 A 09-10-1997 EP 0904289 A 31-03-1999 WO 9737230 A 09-10-1997 US 5863762 A 26-01-1999 CA 2250121 A 09-10-1997 EP 0894269 A 03-02-1999 WO 9636707 A 21-11-1996 IT RM950314 A 18-11-1996 AU 5777696 A 29-11-1996 EP 0826040 A 04-03-1998