JPH06501844A - candida control - Google Patents

Abstract

(57) [Summary] This bulletin contains application data before electronic filing, so abstract data is not recorded.

Description

[Detailed description of the invention] candida control field of invention The present invention relates to diagnostic, research reagents and treatments for Candida infections. More details in the book The invention relates to antisense oligonucleotides that interact with specific Candida senger ribonucleic acids. Concerning gonucleotides. In particular, antisense oligonucleotides Bulin, actin, chitin synthase and aspartic protease proteins is designed to hybridize with Candida mRNA encoding. others The antisense oligonucleotide of liposomal L25 protein, translation elongation Factors 1 and 2 (TEFl and TEF2), b subunit of ATPage and cytochrome P450 lanosterol 14ff-demethylase (LLAI ) designed to specifically hybridize to Candida lllRNA encoding be done. These oligonucleotides alter the activity of Candida RN^ or DNA. was found to lead to the modulation of Candida infection. relaxation and produce a therapeutic effect.

Background of the invention Opportunistic infections in immunocompromised hosts are a significant source of increased mortality and morbidity A case of Candida species are the most common species, Candida albicans species ((an dida albicans), but is the most common fungal pathogen in infected Among the individuals, Candida tropicalis, Candida krusei (Krusei), Candida glabrata (Krusei), Candida glabrata Plata: Torulopsis glabrata) and Candida parabusi parapsilosis was also found. Candida is found in various It is a cause of internal infection. A general review of the types and severity of Candida infections Meunier (Meunier, F6.) Eur! , Cl1n, Micr obiol, Infect, Dis, 8: 438-447 (AW9) Or Raden (R) adentz, W,, )! , A11. Acad, Derm, 20 :989-1003. Cancer patients, especially leukemia patients, are at risk for Candida infections. At high risk. Candida infestations are detected at autopsy in up to 30% of all leukemia patients. Evidence of entry will be shown. Cancer patients with various solid tumors are also at risk for opportunistic infections. Ru. Improvements in cancer treatment with higher doses of surgery and use of novel immunotherapies The result is an increase in the number of non-terminally ill people who are infected with Candida and require treatment. occurred.

Another group at risk for candida and opportunistic infections is the AIDS population. Ru.

Candida is a problem in oropharyngeal infections in people with AIDS. burn victim , I, V drug users, those with catheters, and premature neonates are all included in the category. There is a suspicion that the infection was caused by the virus.

Cancita is also a problem in non-immunocompromised hosts. In normal adult women, Canninda is the cause of vulvovaginitis. The overwhelming majority of yeast infections in the vulva are mosquitoes. It is an isolate of Candida albicans.

This problem is exacerbated by pregnancy and the use of contraceptives or in disease states. The use of biological agents is necessary, all of which increase the likelihood of Candida infection. Ru.

Several drugs are currently in use to treat Candida infections. Amphotericin B is generally considered to be the standard treatment for systemic infections of Cancita.

However, antisense B has many severe side effects, some of which are causes definitive damage to the patient's liver and kidneys. Additionally, amphotericin B has limited efficacy and treatment does not always eliminate the infection. Hence , is effective in suppressing Candida infections but does not cause toxic side effects in the host. There is a great demand for drugs. Antisense oligonucleotides can cause candida It has great promise as a therapeutic agent for cancer. Previously with antisense oligonucleotides There have been attempts to suppress candida. Therefore, oligonucleotides effective for therapeutic use There continues to be a long-standing need for the design of leotide analogs.

Purpose of invention The purpose of the present invention is to inhibit the function of Candida menssencher RNA. oligonucleotides and oligonucleotides that can hybridize with An object of the present invention is to provide oligonucleotide analogs.

In addition, Candida albicans can be detected through antisense interaction with fungal Metsenger RNA. It is further desirable to provide oligonucleotides and analogs that can modulate the expression of It is a purpose.

A further object of the present invention is to provide diagnostic and therapeutic methods for Candida in animals. That is. Detects the presence or absence of Candida in a sample suspected of containing Candida It is a further object of the present invention to provide methods, materials and kits for the methods.

Novel oligonucleotides and oligonucleotide mimics may be used for other purposes of the invention. be.

These and other objects will be apparent to those skilled in the art from this specification and the appended claims. Let it be obvious.

Summary of the invention According to the present invention, it is possible to specifically hybridize with at least a portion of RNA derived from Candida. Oligonucleotides and oligonucleotide analogs are provided.

Oligonucleotides and oligonucleotide analogs are preferably Candida RN Designed to combine directly with A.

This relationship is commonly referred to as "antisense." Oligonucleotides and oligonucleotides Ligonucleotide analogs can inhibit the function of RNA - it affects its proteins any necessary for its translation, or its translocation into the cytoplasm, or its entire biological function. Any other activity. RNA can perform all or part of its functions Absence causes defects in parts of the genome that control the normal life cycle of the fungus. Rub.

Regarding antisense oligonucleotide attack, specific parts of Candida RN^ It has been found preferable to target β-tubulin, aspartic acid Genes encoding protease, actin and chitin synthase were used in this study. It has been found to be particularly useful. Ribosomal L25 protein, translation elongation factor 1 and 2 (TEFI and TEF2), the b subunit of ATPage and Cytochrome? 45G lanosterol 14a-demethylase (LIAI) Also particularly useful are genes encoding. mRNA associated with these proteins It is expected that inhibiting the translation of Candida will be useful in the treatment of Candida infections.

Oligonucleotides and oligonucleotides that allow animals to hybridize with fungal nucleic acids Provided is a method of modulating Candida infection comprising contacting with a leotide analog be done. Oligonucleotides or analogs β-tubulin, aspartic acid mRNA encoding protease, actin and chitin synthase proteins It is preferable that it can be hybridized with. Oligonucleotide or analogue beam Bosomal L25 protein, TEFI and TEF2, the b subunit of ATPase It is also possible to hybridize with Knit and cytochrome P450 LIAI. It is preferable.

IIFI of drawings! explanation Figure 1 shows the β-tube of Candida albicans (Ia albicans) This is the sequence of the phosphorus gene.

Figures 2A and B show oligonucleotide injections of 05 (^) and 1.0 μM (B). The effects of antisense oligonucleotides on Candida germ tube formation in This is a graph.

Detailed description of the invention Antisense oligonucleotides have become important therapeutic agents for many human diseases. have good prospects. Oligonucleotides are defined by Watton-Crick base pairs specifically for the complementary sequence of either the pre-mRNA or the mature mRNA^ It binds and blocks the flow of genetic information from DNA to proteins. Many (recent In research, antisensor as a biochemical tool to study target proteins. The utility of oligonucleotides has been demonstrated. Rothenbarb et al. enberg et al.) Synthesis and enhancement of enzyme-resistant oligonucleotides The potential of oligonucleotide analogues for cellular uptake now makes therapeutic use possible. It is possible to consider the use of antisense oligonucleotides as a new form of be.

Regarding treatment, animals suspected of having a Candida infection should be treated with the oligonucleotides of the present invention. or oligonucleotide analogues. skilled in the art Those skilled in the art can readily determine optimal dosages, dosing methods and repetition rates. such treatment is generally continued until a curative effect or until a reduction in disease status is achieved. It will be done.

Differences in Candida DNA from different species as well as from different types of the same species is expected to exist. For example, the area of various Candida species is serve qualitatively the same functions, as well as interference with the expression of genetic information in different species. It is thought that similar results will be produced. There are undoubtedly differences in nucleotide sequences between species. This is true even if there exists.

Accordingly, the nucleotide sequences described herein are not depicted with respect to the specific species described. It will be understood that Homologues for different species of Candida or analogous arrangements are specifically intended to be included within aspects of the present invention. .

The present invention provides oligonucleotides used for antisense inhibition of the function of Candida RN^. Using cleotide and oligonucleotide analogs. In the context of the present invention , "oligonucleotide" is a polynucleotide formed from naturally occurring bases. It is a pentofuranosyl group linked by a natural phosphodiester bond. . The term refers to naturally occurring species or naturally occurring subunits or effectively refers to a synthetic species formed from close homologs of.

The term "oligonucleotide analog" used in the context of the present invention is , a part that functions similarly to an oligonucleotide but does not exist in nature say. For example, oligonucleotide analogs may have modified sugar moieties or internal-sugar linkages. It may have. Examples of these include phosphorothioates and other sulfur-containing Examples include those known in the art. According to some preferred aspects If at least some phosphodiester bonds in the oligonucleotide are active modulates the ability of the composition to penetrate into areas of the cell where the RNA or DNA is present. Replaced with a structure that functions to enhance. Such substitutions are phosphodiester methylphosphonate linkage or short chain alkyl, or cycloalkyl structure. Preferably, it consists of a structure. According to another preferred embodiment, the phosphodiester linkage At the same time, structures that are substantially non-ionic and non-chiral, or chiral and is substituted with other structures that are enantiomerically specific. Those skilled in the art will be able to carry out the invention. You would be able to select that pond combination to use.

Oligonucleotide analogs include species that contain fewer (and some modified base forms) Good too. For example, such use of purines and pyrimidines other than naturally occurring Can be used. Similarly, the modification on the pentofuranosyl moiety of the nucleotide subunit is This may occur as long as it adheres to the essential teachings of the invention.

Such analogs can be synthesized from natural oligonucleotides (or synthesized along natural systems). is best described as being functionally internally interchangeable with However, it has one or more differences from the natural structure. all such Analogs can be used according to the invention as long as they can effectively hybridize with Candida RN^. Included. The oligonucleotides and oligonucleotide analogs of the invention are It consists of about 3 to about 50 nucleobase units. Such oligonucleotides and similar Preferably, the analog has about 8 to about 25 nucleobase units, and even more preferably about 12 to about 25 units. Layer preferred. It is understood that a subunit or nucleobase unit is , with adjacent subunits via phosphonoester bonds or other bonds, as appropriate. It is a combination of bound base-sugar.

Oligonucleotides and analogues used in the present invention are conveniently and routinely synthesized into solid phase. It is created through the technology of creation. Equipment for such synthesis is available from Abride Bio. It is sold by several vendors including Systems. For such a synthesis Other means can also be used. However, the actual synthesis of oligonucleotides is It is within the skill of the user. Like phosphorothioates and alkylated derivatives The use of similar techniques to prepare oligonucleotide analogues is also well known. It is.

According to the present invention, those skilled in the art will understand that Metsenger RNA uses a three-letter genetic code. In addition to the information that encodes the protein that such a person knows, Untranslated region, 3-untranslated region, 5'cap and intron/exon joining region can be combined to form a region such as bonucleotides and also contain bonucleotides. You will understand that. That is, oligonucleotides prepared according to the present invention and and oligonucleotide analogues as well as informational oligonucleotides. Bind or target all or part of the nucleotide. According to a preferred embodiment , oligonucleotides or analogs at transcription start sites, translation start sites, 3'-untranslated specifically hybridizes with indolone/exon junctions or sequences in the translation region. Wear.

According to the invention, the oligonucleotide is specific to at least a nucleic acid portion of Candida. can be hybridized to. According to a preferred embodiment, the nucleic acid portion contains β-tubulin. , actin, chitin synthase and aspartic protease proteins. Contains the part of the mRNA that encodes. According to another preferred embodiment, the nucleic acid portion includes a bridge. 125 proteins, TEFl and TEF2, b subunit of ATPase Contains the part of the engineer A that coats the cut and Nodochrome P450 LIAI. . Oligonucleotides or analogues consisting of the corresponding sequences, or parts thereof, are Useful in inventions. That is, the oligonucleotides and oligonucleotides of the present invention Creotide analogs can hybridize with Candida metsenger RNA. It is designed to be Such hybridization, when achieved, interferes with the normal function of the trigger RNA and causes a loss of its usefulness to the fungus. Rub. Functions of the mensenger RN^ that are interfered with include, for example, protein translation. transfer of RNA to the place where it is to be processed, actual translation of protein from RNA, Isong and other prosenongs, and possibly by RNA. Even in the case of distinct catalytic activity obtained, all survival functions are mentioned. RN8 The overall effect of such interference with Candida's ability is to eliminate the benefits of RNA for Candida. resulting in the experience of interfering with the expression of the genome as a whole. the Such interference is generally fatal to fungi.

1ffll is β of Candida albicans - Tubulin gene sequence. β-tubulin gene of Candida albicans The gene sequence is known. Smith et al. Gene, 63:53-63 (1988). The gene sequences of these Candida albicans Are known. Au-Young et al., Molecula r　Microbiol.

Gong, 4:197-207 (1990). Actin genetics in Candida albicans The child is well known. Losbergeret al. ) Nucl, Ac1d, Res, 11:9488 (1989). Kan W da The asparatyl brotinase gene of C. albicans was developed by Lott et al. al, , ) by Nucl, Ac1d Res, , 17:1779 (1989). Candida albicans cytochrome P4 50LIA1 sequence was prepared by Lai et al., Nucl, Ac1 d, Res, , 17:804 (P 989). Candida albicans elongation factors TEFI and T EF2 is from Sundstram et al., J. Bacteriol, 172:2036 (1990) ■ Ru. The sequence of the ribosomal L25 gene is Torulopsis glabrata) and Candida eutelis (C andida utilis). long e t al, , ) Nucl, Ac1d Res, , 18: 1888 ( 1990); Wow Woodt et al., Curr, Genet, 12:193 (1 987). Candida tropicalis sky The gene sequence for ATPase subunit b was published by Gu et al. ,.

) Nucl, Ac1dRes, 18 Near 446 (1990). ing.

Oligonucleotides or analogs useful in the invention may have one of these sequences or consists of those parts. For example, any of these oligonucleotides described above (or analogues thereof), or as those skilled in the art would prefer for fungal infection modulation. Use any similar nucleotide that can be prepared from knowledge of the new antisense target. It is preferable that

The oligonucleotides and oligonucleotide analogs of the invention can be used for diagnostic, therapeutic and and research reagents and kits. Regarding therapeutic use, oligonucleotides Candida and oligonucleotide analogs can be administered to animals suffering from Candida infections. Wear. It is generally preferred that therapeutic agents of this sequence be administered locally or intra-injury. Yes. Other forms of administration may also be effective, such as intradermal or intramuscular. Inside the suppository It is currently believed that its inclusion would be very useful. Oligonucleus of the present invention Reotide and oligonucleotide analogues can also be used effectively as prophylactic agents It seems that it will be. They are used as coatings on things like condoms, for example. This can be achieved by administering medication. In some embodiments, a pharmaceutically acceptable carrier may also be used. It is preferable if

The invention is also useful in diagnosis and research. Oligonucleotides of this sequence and oligonucleotide analogs hybridize with Candida-derived nucleic acids. , sandwich and other assays can be constructed to take advantage of this fact. . When oligonucleotides or oligonucleotide analogs and Candida are present The provision of means for detecting hybridization of suspected samples is routinely It can be achieved. Such supplies may include enzyme-linked, radiolabeled or other suitable detection methods. There is a system. Quints that detect the presence or absence of Candida can also be prepared.

Some preferred embodiments of the invention are illustrated by the following examples. for modulation The target mRNA^ is Candida β-tubulin, actin, chitin synthase and and aspartic protease proteins. Other suitable mRNA Targets aliposome L25 protein, translation elongation factors 1 and 2 (TEFI and TEF2), ATPase b subunit and chromochrome P450 lanos terol 14a-demethylase (LIAI). Those skilled in the art will appreciate that the present invention It will be understood that it has general, but not limited, application. These candida Suppression of RNA5 is useful in disease treatment! expected to have significant therapeutic benefits. An assay or assay system is needed to evaluate the effectiveness of the composition.

The following description includes some non-limiting examples of the invention! Figure.

Example Example 1 β-tubulin, actin, chitin synthase and aspartic acid protea Fern β-tubulin, aspartic protease, actin and chitin synthesis A series of antisense oligonucleotide sequences complementary to the mRNA of the enzyme chosen. These are shown in Table 1.

Antisense first nogonucleotide compound targeting Candida albicans Sequence (5°-3°) Target RNA1275 CAA Trr CTCTCA Ding^CTrCTA Tube 1 Non Translation Start Section 1276 CCG AACAT A CAA m CTCTC tube 1 non 5° splice junction intron 1 1277 CAA AACCACTTrA (T person TAT Tr tube 1J) Splice branch, intron 1 1278 ^AAMτT correction^G TAA AAT CA Tubulin sp Rice branch 7-point intron 2 1279 CTA AAA AAA AGCGCA AM GC Chi, j IJ 3° splice junction intron 1 1280 TrCCCA AAA (1: CAn; CACCCT tube 1 no. 3゛Splice junction intron 2 12111 ATG ATA ACT ccATCA TGT TC7 Sparagi Translation initiation protease 1212 cc^^cc ATr CCCGTG TGCGC7 Subaragic acid 585th protease 12113 AACjLAT ACCTAA ACCTTCCk 7 Suharagi Transcription termination protease 1284ACCACCGTCCAT TrT GAA TC actin transcriptional activation First 1285, TrA AAA CAT ACA CCcTCCA 7cutin 5 ゛Splice site 12! +7 TTGTC[; Chin synthase Start of translation 1288 cTcT8τG TCATG TTC person Wakitin synthase second frame ■et1289 TrTAGC-CT AACA TG ACCAC chitin synthase Termination of translation Cancita albicans is a sabro Yeast nitrogen base with dextrose broth (Difco) or glucose Grow in standard broth like medium. Kanshita is 50s! ! antisense o Grow in 1 ml solution of oligonucleotide and its 1/2 log dilution. do. There will be triplicate examiners for each dose. Cancita growth inhibition is r, c5 is expected to occur with 1-10μ oligonucleotide compounds.

Example 2 Synthesis and Characterization of Oligonucleotides and Analogs Unmodified DNA Oligonucleotides Reotide was used as standard using an automatic DNA synthesizer (Abride Biosystems 380B). It was synthesized by oxidation with iodine using phosphoramidite chemistry.

β-cyanoethyl diisopropyl phosphoramidite abride biosystem (Hoster City, California). Phosphorothioate oligonucleotide For cleotides, a stepwise change in phosphite bonds (thiati 3H-1,2- in acetonitrile for standard oxidation (on) It was replaced with a solution of benzodithiol-3-one 1°1-diotinto. Chejo The waiting cycle time was increased to 68 seconds, and then the capping step was performed.

Cleavage and deblotting from controlled pore glass columns (Applied Biosystems) After packing in concentrated ammonium hydroxide at 55°C for 18 hours, the oligonuclear Reotide was purified by precipitation twice with 0.5 M NaCl in 2.5 volumes of ethanol. did. Analytical gel electrophoresis was performed using 20% acrylamide, 8M urea, 45mM Tris- The test was carried out using boric acid buffer, pa 7.0. Oligonucleotides and their phosphorothio More than 80% of the ate analog was a full-length substance as judged by electrophoresis.

Example 3 Germ tube assay for antisense oligonucleotide inhibition of Candida germ tube The development of hyphae is an early stage of hyphal formation, but due to the influence of macrophages considered important for escape. Drugs that inhibit intracellular germ tube formation can effectively promote host defense against Candida infection. Buanwout et al. (V an’t 1out et al, , J, Antimicrob , Chemotherapy, 25:803 (19X0).

Cantuta albicans contains 0.15% asparagine and 2% dextrose. Supplemented yeast nitrogen source basal medium components (Defco Laboratories, Detro, Missouri) Grow the seeds in a container for a day and a night. Cells were pelleted and washed twice with IXPBS. Ru. Add 2% glucose in DMEM to the final pellet to analyze acute effects. Add and resuspend to 5x105 cells/ml. This suspension of 200 μm Add the solution to the wells in a 96-well microtiter plate and add the oligonucleotide Add Tido at desired concentration. Incubate the plate for 1 hour at 37°C under 5% CO□. cuvate. At the end of the incubation, remove 0 glutaraldehyde. ．． Add to 5% and cool the plate to 4°C.

After examining the cells under a microscope and counting the three fractions, the percentage of total cells with germ tubes formed was determined. Measure the rate.

To analyze effects on long-term exposure, cells were exposed to 0.15% asparagine. and suspended in YNB containing 2% dextrose to which oligonucleotides were added. I can do it. Incubate the plate for 4 hours at room temperature, then pellet the cells. Wash with none PBS. The final pellet contained 2% glucose and the new oligo Resuspend in DMEM supplemented with nucleotides. Cells were then placed under 5% CO□. Incubation at 37°C and development as assayed for short-term effects as described above. Perform the bud tube assay.

Oligonucleotides tested in germ tube assays for inhibition of C. albicans The cleotide analogues are shown in Table 2. Table 2 [Place number: 5---------Ichidai-----3" Emperor n Xi 1 22 14 TCT TGT CGA TAA TAT TACCA Chitin synthase ＾UG　P-0#2216　P-3 22215CAA TTT CTCTCA TAG TrCTA J-tube Lin AUG P=OI2217 P=S 3 2754 TC"A C'rG to AT C; GA GCCATr TrC Ribosome L25^UG P=042839CACTCC; ATG CACC CA TTT T to T Ribosome L25^UG P=0#2845 P=S 5 2938 TGT TGT GCA TAA TAT TACCA Chitin Synthetic enzyme A [IG P=S73156Trr ACCCAT GAT TGA TTA TAT TEFI Oyohi TEF2 AUG p,, □J3122 P =S 8 3121 TCA CTC: GAT G to A GCCATT’! Til : Ribosome L25 AUG P=Og 3152 TGA CAT CAT CAAπG ATG 8C to ATPase sub-L knee/) b AUG P =0#3125 P=S 10 3150 cTG cλT AAT ATTATCATCAjlα Kichi Synthase AUG P=S113151AGCCAT ATT GA (: TT Enter τGA TcT cytochrome P 450 LIAIP Tomi S 12 1049 GCCGAG GTCCAT GTCcrx ccc control- ■SV UL13 P=0' 1082 P=S Figures 2A and B are specifically linked to the chitin synthase gene of Cancita albicans. hybridizing antisense oligonucleotides (phosphonic esters and phosphonates) Results of a time-course analysis comparing the effects of Holothioate analogs) with several controls. represents the fruit. l5IS 2214 (SEo 10 NO: 1) is a chitin synthase Revised SJS, a phosphodiester oligonucleotide targeting the AUG region 2216 (SEQ IDN0:1) Hal5IS 2214 nophospho o thio x- SIS 1049 and 15IS 1082, which are analogues of (SEQ ID NO: 12) Translation of the mRNA product of the herpes simplex virus UL13 gene phosphodiester and control sequences that hybridize to the initiation codon, respectively. It is a phosphorothioate analog. "Control" refers to untreated cells. 05 and 1. Germ tube assay with two oligonucleotide doses of H.ii is shown in Figure 2 (A ) and FIG. 2(B), respectively. In both administrations, l5IS 221 6 (phosphorus that hybridizes with mRNA encoding kanshita chitin synthase) (rothioate oligonucleotide analogues) inhibit germ tube formation in Cancita as compared to other compounds. It was shown that it was greatly inhibited compared to the physical object.

Sequence list (1) General information: (i) Applicant Hoke Glenn C. Eckerday Bisoto J, (Ecker, David J,) (Hill) Name of invention Kanshita suppression of (i) Number of arrays: 12 (i+) Address: (A) Address: Woodcock, Onon Yuban, Carla McKeevinotsu, AND1 Norris (Woodcock Washburn Kurtz Mack ievicz & Norris) (B) One Liberty Brace-4 6th floor (One Liberty Place) (C) City name: Philadelphia (D) State name: Pennluvenia (E) Country name: United States (F) Postal code 19103 (V) Computer-readable format.

(A) Medium/small disk, 35 inches, storage device 1.44Mb (B) Computer IBM PS/2 (C) operating system: PC-DO3 (D) Software, WORDPERFECT 5.0 ．． 0) (vi) Current application materials (A) Application number n/a (B) Filing date 9 Together with this specification (C) Classification.

(vi) Patent attorney/agent information: (A) Name, Licata, Jane M. (B) ) Registration number: 32.257 (C) Inquiry/case reference number: l5IS-0432 (ix) Communication information.

(A) Telephone: (215) 568-3100 (B) Telefax: (215) ) 56g-3439 (2) Information on SEQ ID NO: 1・ (i) Features of array (^) Length 20 base pairs (B) Seed: Nucleic acid (ii) Molecule type: other nucleic acids (i) No hypothesis m) anti-sense positive (c) Sequence description Sequence number I T G T T G T CG A T, A A T, A T T A C CA20(2) Information on sequence number 2 (i) Features of array (A) Length 20 base pairs (B) Type, nucleic acid (C) Chain = 1 chain (D) Topology/straight line (i) Molecule type: other nucleic acids (i) Hypothesis, none i) anti-sense positive (m) Sequence description: Sequence number 2 CAATTTCTCT CATAGTTCTA 20 (2) Information on sequence number 3: (i) Features of array (A) Length 21 base pairs (B) Type: Nucleic acid (C) Chain, 1 (D) Topology two straight lines (fi) Molecule types/other nucleic acids (i) Hypothesis: None Xl) Sequence description: Sequence number 3 TCACTGGATG GAGCCATTT C21 (2) Information on SEQ ID NO: 4 : (1) Characteristics of array (A) Length = 21 base pairs (B) Type: Nucleic acid (C) One chain (D) Topo day no. straight line (Ii) Nucleic acid with molecular type ability (i) No hypothesis ff) anti-sense positive (Xl) Description of sequence゛Sequence number 4 CACTGGATGC, CCCATTTTG T21 (2) Information of SEQ ID NO: 5 Information (1) Characteristics of array (In) Length 21 base pairs (B) Type, nucleic acid (C) Chain: 1 (D) Topolon-1II line (li) Molecule type, Ike's nucleic acid (i) Hypothesis/None ff) Anti-sense: Correct (Sword) Sequence description: Sequence number 5 CTCATAGTTCTATAATGTTG A 21 (2) Information on SEQ ID NO: 6 : (1) Characteristics of array (A) Length = 20 base pairs (B) Type: Nucleic acid (C) Chain = 1 chain (D) Topology two straight lines (i) Molecule type: other nucleic acids (i) Hypothesis/None quit) anti-sense: positive (=) Sequence description: Sequence number 6 TGTTGTGCAT AATATTACCA 20 (2) Information on SEQ ID NO: 7.

(i) Features of array (A) Length: 21 base pairs (B) Type of nucleic acid (C) 1 chain (D) Topology straight line (ii) Nucleic acids with molecular type capabilities (i) No hypothesis W) anti-sense, correct (J) Sequence description Sequence number 7 TTTACCCATG ATTGATTAT, 8 T 21 (2) SEQ ID NO: 8 information (i) Features of array (A) Length 21 base pairs (B) Type/nucleic acid (C) Chain, 1 (D) Topology straight line (fi) Nucleic acid with molecular type ability (i) No hypothesis y) Anti-sense positive (n) Sequence description, SEQ ID NO. 8 TCACTGGATG GAGCCATTT G 21 (2) Information of SEQ ID NO: 9 News.

(i) Features of array (A) Length 21 base pairs (B) Type of nucleic acid (C) 1 chain (D) Topology tl! 1! (j) Nucleic acid with molecular type ability (i) Hypothesis/None ff) anti-sense positive (J) Sequence description, SEQ ID NO. 9 TGACATGATG A, ATGGATGAC, 821(2) SEQ ID NO: 10 information.

(1) Characteristics of array (A) Length/21 base pairs (B) Type: Nucleic acid (C) One chain (D) Topology straight line (6) Molecule types, other nucleic acids (i) Hypothesis/None ff) anti-sense, positive (r) Sequence description Sequence number 10 GTGCATAAT, A TTACCATCAA T 21 (2) SEQ ID NO: 11 information (1) Characteristics of array (A) Length 21 base pairs (B) Type of nucleic acid (C) Chain. 1 piece (D) Topology/straight line (D number of molecule types of nucleic acids (i) No hypothesis (quit) anti-sense, positive (J) Sequence description: Sequence number 11 AGCCATATTG AGTTATGATCT 21 (2) Information of SEQ ID NO: 12 Information: (1) Characteristics of array (A) Length, 21 base pairs (B) Type, nucleic acid (C) Chain: 1 (D) Topology: straight line (Yu) Molecule type: other nucleic acids (i) Hypothesis: Coral W) Anti-sense: Correct (f+) Sequence description, SEQ ID NO: 12 GCCGAGGTCCATGTCGTACG C21no, 1 1201　TTrAAGAAAA　TTGGCAGTCA　ATrTGGTrC (: ATTCCCAAGA TrACATTTCT1251 TTATC;C :TT to G TTATG (TCCA rrcAcrrcrATGGGTrCTA A　Arcrrrrcac＾1851　G’f’TCACTTCT　AA’KAT AATrA TGGmmT TGGmAG There is no midsummer in the content written by Bee) FIG, 2A Procedural amendment

Claims (31)

[Claims] 1. Candida β-tubulin, actin, and chitin synthase , aspartic protease, translation elongation factor 1, translation elongation factor 2, ribosome L25 protein, ATPase b subunit or cytochrome P450 A small amount of mRNA encoding the lanosterol 14α demethylase protein. An oligonucleotide or oligonucleotide that can specifically hybridize with a moiety Creotide analog. 2. mRNA transcription start site, translation start site, intron/exon junction, or or a claim capable of specifically hybridizing with at least a portion of the 5' cap region. Item 1. Oligonucleotide or oligonucleotide analog according to item 1. 3. An oligonucleotide according to claim 1 in a pharmaceutically acceptable carrier. or oligonucleotide analogs. 4. The oligonucleosome of claim 1 having from 4.5 to about 50 nucleobase units. tide or oligonucleotide analogs. The oligonucleosome of claim 1 having from 5.8 to about 25 nucleobase units. tide or oligonucleotide analogs. 6. The oligonucleotide of claim 1 having from 12 to about 25 nucleobase units. Otide or oligonucleotide analogs. 7. At least some of the linking groups between the nucleotide units of the oligonucleotide are The oligonucleotide or oligonucleotide of claim 1 comprising a sulfur-containing species. de analogue. 8. At least some linking groups between the nucleotide units of the oligonucleotide The oligonucleotide or oligonucleotide of claim 1 comprising a phosphorothioate moiety. Gonucleotide analogs. 9. The ointment according to claim 1, wherein the mRNA encodes β-tubulin protein. Ligonucleotides or oligonucleotide analogs. 10. Candida RNA and sequence; [sequence available], [There is an array], [There is an array], [There is an array], [There is an array], [There is an array], [There is an array], [There is an array], [There is an array], [There is an array], [There is an array], [There is an array], [There is an array], [There is an array], or [There is an array]. an oligonucleotide or oligonucleotide capable of specifically hybridizing with at least a portion of or oligonucleotide analogs. 11. The oligonucleotide of claim 10 in a pharmaceutically acceptable carrier. oligonucleotide analogs. 12. at least some linking groups between the nucleotide units of the oligonucleotide 11. The oligonucleotide or oligonucleotide according to claim 10, wherein comprises a sulfur-containing species. Otide analogue. 13. at least some linking groups between the nucleotide units of the oligonucleotide The oligonucleotide according to claim 10, wherein comprises a phosphorothioate moiety or Oligonucleotide analogs. 14. A method for treating Candida infection, the method comprising: Candida β-tubulin, actin, and chitin synthesis enzyme, aspartic protease, translation elongation factor 1, translation elongation factor 2, ribosomal ATPase b subunit or cytochrome P4 A small amount of mRNA encoding the 50 lanosterol 14α demethylase protein. an oligonucleotide or oligonucleotide capable of specifically hybridizing with at least a portion of the A method comprising contacting with a gonucleotide analogue. 15. Oligonucleotides or oligonucleotide analogs can be used to initiate transcription of mRNA. start site, translation start site, intron/exon junction, or 5' cap region 15. The method of claim 14, wherein the method is capable of specifically hybridizing with at least a portion of. 16. The oligonucleotide or oligonucleotide analog is pharmaceutically acceptable. 15. The method of claim 14 in a carrier comprising: 17. 5 to about 50 oligonucleotides or oligonucleotide analogs The method according to claim 14, having a nucleobase unit of. 18. 8 to about 25 oligonucleotides or oligonucleotide analogs The method according to claim 14, having a nucleobase unit of. 19. The oligonucleotide or oligonucleotide analog has 12 to about 25 15. The method according to claim 14, wherein the method has nucleobase units. 20. Nucleotide units of oligonucleotides or oligonucleotide analogs 15. The method of claim 14, wherein at least some of the linking groups between include sulfur-containing species. 21. Nucleotide units of oligonucleotides or oligonucleotide analogs Claim 1, wherein at least some of the linking groups between the How to put it on. 22. 2. The mRNA according to claim 1, wherein the mRNA encodes β-tubulin protein. Method. 23. The infection is Candida albicans, Candida tropi calis. Candida Krusei. Torulopsis grab A claim that is an infection of C. rata or Candida parapsilosis. 14. The method described in 14. 24. A method for modulating the activity of Candida, which can be used to treat patients with Candida infection. Then arrange the suspected animals; [There is an array], [There is an array], [There is an array], [There is an array], [There is an array], [There is an array], [There is an array], [There is an array], [There is an array], [There is an array], [There is an array], [There is an array], [There is an array], [There is an array], or [There is an array]. an oligonucleotide or oligonucleotide comprising at least a portion of one of A method comprising contacting with an otide analogue. 25. specifically hybridizes with RMA of Candida, and An oligonucleotide or oligonucleotide comprising at least a portion of one of the sequences defined in Table 2. or oligonucleotide analogs. 26. An oligonucleotide according to claim 25 in a pharmaceutically acceptable carrier. oligonucleotide analogs. 27. at least some linking groups between the nucleotide units of the oligonucleotide 26. The oligonucleotide or oligonucleotide of claim 25, wherein the oligonucleotide comprises a sulfur-containing species. Otide analogue. 28. at least some linking groups between the nucleotide units of the oligonucleotide 26. The oligonucleotide of claim 25, wherein comprises a phosphorothioate moiety or Oligonucleotide analogs. 29. A method for modulating the activity of Candida, the method comprising: Animals suspected of having an infection are treated with at least one of the sequences defined in Table 2. therapeutically useful oligonucleotides or oligonucleotide analogs containing a portion of A method consisting of contacting with an effective amount. 30. Nucleotide units of oligonucleotides or oligonucleotide analogs 30. The method of claim 29, wherein at least some of the linking groups therebetween include a sulfur-containing species. 31. Nucleotide units of oligonucleotides or oligonucleotide analogs Claim 29, wherein at least some of the linking groups between Method described.