USE OF THE GENES IN THE HOG, Ras AND cAMP PATHWAY FOR TREATMENT OF FUNGAL INFECTION

Abstract

Provided herein are uses of genes for HOG, Ras and cAMP signal transduction pathways to treat fungal infection. To regulate the HOG pathway of Cryptococcus neoformans , roles of SSK1, TCO2, SSK2, PBS2, HOG1, ENA1 and NHA1 genes were investigated to find that a biosynthesis level of ergosterol is increased when these genes are inhibited. When the genes are inhibited, a large amount of ergosterol is distributed on a fungal cell membrane. Accordingly, since there are many working points of an ergosterol-binding antifungal agent, an efficiency of the ergosterol-binding antifungal agent can be considerably improved. To regulate the Ras and cAMP pathways of Cryptococcus neoformans , roles of RAS1, RAS2, CDC24, GPA1, CAC1, ACA1, PKA1, HSP12 and HSP122 genes were investigated to find that a sensitivity to a polyene- or azole-based drug is increased when these genes are inhibited. Therefore, an antifungal pharmaceutical composition including an inhibitor against the gene or protein encoded by the same can be used as an excellent combined antifungal agent which can reduce a conventional amount of an antifungal agent used and increase an efficiency.

1 . A method of treating fungal infection, comprising:
 injecting an effective amount of an inhibitor against at least one protein selected from the group consisting of Ssk1, Tco2, Ssk2, Pbs2, Hog1, Ena1 and Nha1 of  Cryptococcus neoformans  into a subject.   
     
     
         2 . The method of treating fungal infection of  claim 1 , wherein the inhibitor against at least one protein selected from the group consisting of Ssk1, Ena1 and Nha1. 
     
     
         3 . The method of treating fungal infection of  claim 1 , wherein an ergosterol-binding antifungal agent or azole-based antifungal agent is sequentially or simultaneously injected with the inhibitor. 
     
     
         4 . The method of treating fungal infection of  claim 3 , wherein the ergosterol-binding antifungal agent is a polyene-based antifungal agent. 
     
     
         5 . The method of treating fungal infection of  claim 4 , wherein the polyene-based antifungal agent is at least one selected from the group consisting of amphotericin B, natamycin, rimocidin, filipin, nystatin and candicin. 
     
     
         6 . The method of treating fungal infection of  claim 5 , wherein the polyene-based antifungal agent is amphotericin B. 
     
     
         7 . The method of treating fungal infection of  claim 3 , wherein the azole-based antifungal agent is at least one selected from the group consisting of ketoconazole, fluconazole, itraconazole and voriconazole. 
     
     
         8 . A method of treating fungal infection comprising:
 Injecting an effective amount of an inhibitor against at least one gene selected from the group consisting of SSK1, TCO2, SSK2, PBS2, HOG1, ENA1 and NHA1 of  Cryptococcus neoformans  into a subject.   
     
     
         9 . The method of treating fungal infection of  claim 8 , wherein the inhibitor against at least one gene selected from the group consisting of SSK1, ENA1 and NHA1. 
     
     
         10 . The method of treating fungal infection of  claim 8 , wherein an ergosterol-binding antifungal agent or azole-based antifungal agent is sequentially or continuously injected with the inhibitor. 
     
     
         11 . The method of treating fungal infection of  claim 10 , wherein the ergosterol-binding antifungal agent is a polyene-based antifungal agent. 
     
     
         12 . The method of treating fungal infection of  claim 11 , wherein the polyene-based antifungal agent is at least one selected from the group consisting of amphotericin B, natamycin, rimocidin, filipin, nystatin, and candicin. 
     
     
         13 . The method of treating fungal infection of  claim 12 , wherein the polyene-based antifungal agent is amphotericin B. 
     
     
         14 . The method of treating fungal infection of  claim 10 , wherein the azole-based antifungal agent is at least one selected from the group consisting of ketoconazole, fluconazole, itraconazole and voriconazole. 
     
     
         15 . An antifungal combined formulation, comprising:
 an inhibitor against at least one protein selected from the group consisting of Ssk1, Tco2, Ssk2, Pbs2, Hog1, Ena1 and Nha1 of  Cryptococcus neoformans ; and   an ergosterol-binding antifungal agent or azole-based antifungal agent.   
     
     
         16 . A method of screening an antifungal agent comprising:
 contacting at least one protein selected from the group consisting of Ssk1, Tco2, Ssk2, Pbs2, Hog1, Ena1 and Nha1 of  Cryptococcus neoformans  with a candidate material;   and   determining whether the candidate material inhibits or stimulates an activity of the protein.   
     
     
         17 . The method of  claim 16 , wherein the antifungal agent is an inhibitor against an Ssk1, Ena1 or Nha1 protein. 
     
     
         18 . A method of screening an antifungal agent comprising:
 contacting at least one gene selected from the group consisting of SSK1, TCO2, SSK2, PBS2, HOG1, ENA1 and NHA1 of  Cryptococcus neoformans  with a candidate material;   and   determining whether the candidate material inhibits or stimulates an activity of the gene.   
     
     
         19 . The method of  claim 18 , wherein the antifungal agent is an inhibitor against an SSK1, ENA1 or NHA1 gene. 
     
     
         20 . A method of treating fungal infection, comprising:
 injecting an effective amount of an inhibitor against at least one protein selected from the group consisting of Ras1, Ras2, Cdc24, Gpa1, Cac1, Aca1, Pka1, Hsp12 and Hsp122 of  Cryptococcus neoformans  into a subject.   
     
     
         21 . The method of treating fungal infection of  claim 20 , wherein the inhibitor is an inhibitor against a Cac1 or Pka1 protein. 
     
     
         22 . The method of treating fungal infection of  claim 20 , wherein a polyene- or azole-based antifungal agent is sequentially or simultaneously injected with the inhibitor. 
     
     
         23 . The method of treating fungal infection of  claim 22 , wherein the polyene-based antifungal agent is at least one selected from the group consisting of amphotericin B, natamycin, rimocidin, filipin, nystatin, and candicin. 
     
     
         24 . The method of treating fungal infection of  claim 23 , wherein the polyene-based antifungal agent is amphotericin B. 
     
     
         25 . The method of treating fungal infection of  claim 22 , wherein the azole-based antifungal agent is at least one selected from the group consisting of ketoconazole, fluconazole, itraconazole and voriconazole. 
     
     
         26 . The method of treating fungal infection of  claim 20 , wherein an inhibitor against at least one protein selected from the group consisting of SSK1, TCO2, SSK2, PBS2, HOG1, ENA1 and NHA1 of  Cryptococcus neoformans  is sequentially or simultaneously injected with the inhibitor. 
     
     
         27 . A method of treating fungal infection, comprising:
 injecting an effective amount of an inhibitor against at least one gene selected from the group consisting of RAS1, RAS2, CDC24, GPA1, CAC1, ACA1, PKA1, HSP12 and HSP122 of  Cryptococcus neoformans  into a subject.   
     
     
         28 . The method of treating fungal infection of  claim 27 , wherein the inhibitor is an against CAC1 or PKA1 gene. 
     
     
         29 . The method of treating fungal infection of  claim 27 , wherein a polyene- or azole-based antifungal agent is sequentially or simultaneously injected with the inhibitor. 
     
     
         30 . The method of treating fungal infection of  claim 29 , wherein the polyene-based antifungal agent is at least one selected from the group consisting of amphotericin B, natamycin, rimocidin, filipin, nystatin, and candicin. 
     
     
         31 . The method of treating fungal infection of  claim 30 , wherein the polyene-based antifungal agent is amphotericin B. 
     
     
         32 . The method of treating fungal infection of  claim 29 , wherein the azole-based antifungal agent is at least one selected from the group consisting of ketoconazole, fluconazole, itraconazole and voriconazole. 
     
     
         33 . The method of treating fungal infection of  claim 27 , wherein an inhibitor against at least one gene selected from the group consisting of SSK1, TCO2, SSK2, PBS2, HOG1, ENA1 and NHA1 of  Cryptococcus neoformans  is sequentially or simultaneously injected with the inhibitor. 
     
     
         34 . An antifungal combined formulation, comprising:
 an inhibitor against at least one protein selected from the group consisting of Ras1, Ras2, Cdc24, Gpa1, Cac1, Aca1, Pka1, Hsp12 and Hsp122 of  Cryptococcus neoformans ; and   at least one antifungal agent selected from the group consisting of a polyene-based antifungal agent, an azole-based antifungal agent, and an inhibitor against at least one protein or gene coding for the same selected from the group consisting of SSK1, TCO2, SSK2, PBS2, HOG1, ENA1 and NHA1 of  Cryptococcus neoformans .   
     
     
         35 . A method of screening an antifungal agent comprising:
 contacting at least one protein selected from the group consisting of Ras1, Ras2, Cdc24, Gpa1, Cac1, Aca1, Pka1, Hsp12 and Hsp122 of  Cryptococcus neoformans  with a candidate material; and   determining whether the candidate material inhibits or stimulates an activity of the protein.   
     
     
         36 . The method of  claim 35 , wherein the antifungal agent is an inhibitor against a Cac1 or Pka1 protein. 
     
     
         37 . A method of screening an antifungal agent comprising:
 contacting at least one gene selected from the group consisting of RAS1, RAS2, CDC24, GPA1, CAC1, ACA1, PKA1, HSP12 and HSP122 of  Cryptococcus neoformans  with a candidate material; and   determining whether the candidate material inhibits or stimulates the activity of the gene.   
     
     
         38 . The composition method of  claim 37 , wherein the antifungal agent is an inhibitor against a CAC1 or PKA1 gene.