Previous Article | Next Article 
Antimicrobial Agents and Chemotherapy, March 1999, p. 589-591, Vol. 43, No. 3
0066-4804/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
Treatment of Murine Fusariosis with SCH
56592
M.
Lozano-Chiu,1,*
S.
Arikan,1
V. L.
Paetznick,1
E. J.
Anaissie,2
D.
Loebenberg,3 and
J. H.
Rex1
Division of Infectious Diseases, Department
of Internal Medicine, Center for the Study of Emerging and
Reemerging Pathogens, University of Texas Medical School, Houston,
Texas 770301; The Myeloma and
Transplantation Research Center, University of Arkansas for Medical
Sciences, Little Rock, Arkansas 772052; and
Schering-Plough Research Institute, Kenilworth, New
Jersey3
Received 1 October 1998/Returned for modification 9 November
1998/Accepted 4 December 1998
 |
ABSTRACT |
Doses of 10 to 100 mg of the azole antifungal agent SCH 56592/kg of
body weight/day were studied in immunocompetent mice as therapy for systemic infection by Fusarium solani.
Treatment was begun 1 h after intravenous infection and
continued daily for 4 or 13 doses. Prolongation of survival and organ
clearance were dependent on both the dose and the duration of SCH 56592 therapy, with the best results seen at 50 and 100 mg/kg/day. The
results at the highest doses of SCH 56592 used (50 or 100 mg/kg/day)
were comparable to those obtained with amphotericin B at 1 mg/kg/day. SCH 56592 has potential for therapy of systemic infections caused by
F. solani.
| |
INTRODUCTION |
Fusarium species,
traditionally considered plant pathogens, are now known to produce
life-threatening infections in patients with compromised immune
responses (13, 24). Of the various species of
Fusarium, the major species isolated from humans has been
Fusarium solani, followed by F. oxysporum and
F. moniliforme (3-5, 8, 10, 14, 18, 20, 22). In
all, as many as 15 different Fusarium species have been
reported as causing infections in humans and animals (8).
Although the status of the host and the degree of tissue invasion
remain the most important factors in predicting the outcome of
Fusarium infections, the availability of effective
antifungal agents is desirable (1). Unfortunately, the in
vitro activity of the available agents is not very good. Polyenes such
as amphotericin B and natamycin have in vitro activity, but their use
is limited by toxicity (1, 17, 19, 21). Flucytosine and the
currently available azoles (fluconazole and itraconazole) have no
meaningful activity in vitro, and resistance is practically universal
(17, 19, 21), although the combination therapy of
flucytosine and ketoconazole has been reported to have activity against
extensive subcutaneous hyphomycosis caused by F. solani
(4). SCH 56592 is a novel triazole antifungal with in vitro
and in vivo activity against a variety of fungi, including
Candida spp., Cryptococcus neoformans,
Coccidioides immitis, Aspergillus fumigatus, and
many other opportunistic yeasts and molds (9, 11, 15, 16, 23). In this work we have studied the activity of SCH 56592 in
the treatment of disseminated fusarial infection in nonneutropenic mice.
(This work was presented in part at the 98th General Meeting of the
American Society for Microbiology, 1998 [10a].)
| |
MATERIALS AND METHODS |
Organism.
A single isolate of F. solani (FS 1184;
Fusarium Research Center Culture Collection, Pennsylvania
State University) was used for all experiments. This organism was
isolated from the blood of a cancer patient with invasive fusariosis.
Stock lyophilized vials from the fungal strain were obtained from the
Fusarium Research Center and kept at 4°C until use
(7). For both susceptibility testing and animal studies, FS
1184 was grown in Fusarium culture medium (1 liter of
5% glucose in water, 1 liter of sterile saline solution [0.85%
NaCl], 2 g of potassium chloride [KCl], and 2 ml of
Tween 80 [polyoxyethylene-20-sorbitan monooleate]) for 72 h. The
conidia were then harvested and prepared as described by Anaissie et
al. (2).
Susceptibility testing.
By using an adaptation of the
microdilution variant of the National Committee for Clinical Laboratory
Standards procedure M27-A (12), FS 1184 was tested two
different times against SCH 56592, fluconazole, itraconazole, and
amphotericin B. Fluconazole was dissolved in water, and itraconazole,
SCH 56592, and amphotericin B were dissolved in dimethyl sulfoxide,
according to the M27-A protocol. The conidial inoculum was adjusted to
80 to 82% of transmittance at 530 nm, by following the methodology of
Espinel-Ingroff et al. (6), to produce a stock inoculum of
0.5 × 106 to 3.0 × 106 CFU/ml. The
MIC for the azoles was defined as the concentration of the well which
visually showed growth that was less than 20% of growth in the control
well after 48 h of incubation. The MIC for amphotericin B was
defined as the lowest concentration at which no growth was visible.
Animal model.
Four- to six-week-old healthy male CF1 mice
with a mean weight of 24 g (Harlan Sprague Dawley, Inc.,
Indianapolis, Ind.) were used. No immunosuppressant agents were used.
The inoculum for infection was prepared by adjusting the organism to a
transmittance at 530 nm of 10%, which produced 1 × 107 to 3 × 107 CFU/ml, and was
quantitated by plating onto Sabouraud dextrose agar. Infection was
established by injecting 0.2 ml of the adjusted inoculum via the tail
vein. After infection, mice were observed twice daily, and animals
exhibiting profound inanition or an inability to reach food and water
were sacrificed. SCH 56592 was prepared for administration by heating
4 g of methylcellulose (Sigma, St. Louis, Mo.) at 80°C for 90 min in 1 liter of distilled water. After approximately 45 min, 5.6 ml
of Tween 80 (Fisher Scientific, Fair Lawn, N.J.) and 9 g of NaCl
(EM Industries, Gibbstown, N.J.) were added. This solution was
autoclaved. SCH 56592 was added after cooling, and the mixture was
stored at 4°C until use. For amphotericin B, a vial containing 50 mg
of amphotericin B USP (Pharma-Tek, Huntington, N.Y.) for injection was
reconstituted per the manufacturer's instructions. SCH 56592 was given
orally by gavage once daily in a final volume of 0.1 ml at daily doses of 10, 25, 50, or 100 mg/kg of body weight. Amphotericin B was administered intraperitoneally once daily at a final dose of 1 mg/kg in
a final volume of 0.1 ml. Treatment with both drugs was begun 1 h
after intravenous infection and continued daily for a total of 4 or 13 doses. Groups of 16 to 20 animals each were treated with either
amphotericin B, SCH 56592, or methylcellulose diluent or were left
untreated. On days 5 and 14 (24 h after the last dose of drug), five
mice from each group were sacrificed for determination of the numbers
of CFU of F. solani per gram in the liver and kidney. This
determination was made by removing and weighing the organs,
homogenizing the organs in 10 ml of sterile saline with a Stomacher 80 (A. J. Seward, UAC House, London, England), and plating
appropriate dilutions on Sabouraud dextrose agar. The plates were
incubated at 35°C for 48 h, and the number of colonies was then
counted. All surviving animals were sacrificed at 21 days after
infection. All animal care procedures were supervised and approved by
the University of Texas
Houston Animal Welfare Committee.
Statistical methods.
Survival times were estimated by the
Kaplan-Meier method and were compared by the log rank test. Organ
clearance results were subjected to analysis of variance. Calculations
were performed by using SPSS for Windows, version 7.5.1 (SPSS, Inc.,
Chicago, Ill.).
| |
RESULTS |
In vitro susceptibility testing results.
The MIC of
fluconazole and itraconazole against FS 1184 was >64 µg/ml, that of
amphotericin B was 1.0 µg/ml, and that of SCH 56592 was 4.0 µg/ml.
These results suggested that FS 1184 was resistant to fluconazole and
itraconazole but potentially susceptible to amphotericin B and SCH 56592.
In vivo study. (i) Survival and mortality.
The effects of
treatment on the survival of mice infected with FS 1184 are shown in
Table 1. In all, three different
experiments, using various combinations of doses and treatment
durations, were performed. Mice treated with the methylcellulose
diluent had the same duration of survival (6.2 to 8.4 days) as
untreated mice (5.9 to 9.5 days). A significant and dose-dependent
effect of therapy on prolongation of survival was apparent for SCH
56592. The most consistently increased survival results were obtained with SCH 56592 at 50 mg/kg/day, suggesting that the peak of the dose-response curve had been attained. The etiology of the slight decrease in survival noted when 4 daily doses of SCH 56592 were given
at 100 mg/kg/day is not clear, and this effect was not observed when 13 doses were given. The results of treatment with SCH 56592 at 25, 50, and 100 mg/kg/day were not statistically different from those produced
with amphotericin B at 1 mg/kg/day.
(ii) Response to therapy.
Data on the reduction in the numbers
of CFU per gram of kidney and liver are shown in Tables
2 and 3,
respectively. Reduction in the numbers of CFU per gram was a function
of the dose and duration of therapy in both the kidney and the liver,
with maximal reductions produced after 13 daily doses of therapy with
SCH 56592 at 50 to 100 mg/kg. In the liver in particular, this duration of therapy resulted in complete clearance of the infecting organism. The organ clearance results after four doses of therapy with SCH 56592 were similar to the clearance seen with four doses of amphotericin B.
| |
DISCUSSION |
SCH 56592 has previously been shown to have excellent activity
both in vitro (9, 16, 23) and in vivo (11, 15,
23) against a variety of fungi, including Candida
spp., Blatomyces dermatitidis, C. immitis, and
Aspergillus spp. In this study, we have shown that SCH 56592 is also active both in vitro and in vivo against F. solani
in an immunocompetent mouse model of systemic fusariosis.
The effect of SCH 56592 in vivo was dependent on both the dose and the
duration of therapy. Optimal effects were produced when 13 doses were
given at 50 to 100 mg/kg/day. This dose optimum is similar to, but
slightly higher than, the optimum dose in other studies with this
compound. Oakley et al. (15) demonstrated that SCH 56592 at
25 mg/kg/day was better than itraconazole or amphotericin B in a
neutropenic murine model of aspergillosis. Lutz et al. (11)
found that doses of 10 and 25 mg/kg/day cured mice infected with
C. immitis. Finally, Sugar and Liu (23) have shown that 25 mg/kg/day sterilized the lungs of mice infected with
B. dermatitidis. Although a slight diminution in survival was noted when four daily doses of SCH 56592 were given at 100 mg/kg/day, this effect was not observed when therapy was given for a
longer period. Thus, the higher doses of SCH 56592 required for
treatment of this difficult infection appeared well tolerated.
Of interest, SCH 56592 appeared to completely clear the infection in
the liver. The liver appears to be preferentially targeted by the
fungus in this model (the numbers of CFU per gram of tissue in the
livers of untreated animals are three- to fourfold higher than those in
the kidneys), thus making this effect even more apparent. The
relatively less effective clearance of the renal infection may be a
function of the pharmacokinetics of SCH 56592, as effective
concentrations of SCH 56592 in the liver are two- to fourfold higher
than those in the kidney (20a).
In conclusion, we found that SCH 56592 increased survival and reduced
organ burdens in immunocompetent mice infected with F. solani. Prolongation of survival and organ clearance were
dependent on both the dose and the duration of SCH 56592 therapy.
Similar results were obtained with amphotericin B. SCH 56592 has
potential for therapy of systemic infections in humans caused by
F. solani.
| |
ACKNOWLEDGMENT |
This work was supported in part by a grant to M. Lozano-Chiu from
the Dirección General de Investigación Cientifica y
Enseñanza Superior of Spain.
| |
FOOTNOTES |
*
Corresponding author. Mailing address: 6431 Fannin,
1728 JFB, Houston, TX 77030. Phone: (713) 500-6755. Fax: (713)
500-5495. E-mail: mchiu{at}heart.med.uth.tmc.edu.
| |
REFERENCES |
| 1.
|
Anaissie, E.,
H. Kantarjian,
J. Ro,
R. Hopfer,
K. Rolston,
V. Fainstein, and G. Bodey.
1988.
The emerging role of Fusarium infections in patients with cancer.
Medicine (Baltimore)
67:77-83[Medline].
|
| 2.
|
Anaissie, E. J.,
R. Hachem,
C. Legrand,
P. Legenne,
P. Nelson, and G. P. Bodey.
1992.
Lack of activity of amphotericin B in systemic murine fusarial infection.
J. Infect. Dis.
165:1155-1157[Medline].
|
| 3.
|
Anaissie, E. J., and M. G. Rinaldi.
1990.
Fusarium and the immunocompromised host: liaisons dangeureuses.
N. Y. State J. Med.
90:586-587[Medline]. (Comment.)
|
| 4.
|
Attapattu, M. C., and C. Anandakrishnan.
1986.
Extensive subcutaneous hyphomycosis caused by Fusarium oxysporum.
J. Med. Vet. Mycol.
24:105-111[Medline].
|
| 5.
|
Collins, M. S., and M. G. Rinaldi.
1977.
Cutaneous infection in man caused by Fusarium moniliforme.
Sabouraudia
12:151-160.
|
| 6.
|
Espinel-Ingroff, A.,
K. Dawson,
M. Pfaller,
E. Anaissie,
B. Breslin,
D. Dixon,
A. Fothergill,
V. Paetznick,
J. Peter,
M. Rinaldi, and T. Walsh.
1995.
Comparative and collaborative evaluation of standardization of antifungal susceptibility testing for filamentous fungi.
Antimicrob. Agents Chemother.
39:314-319[Abstract/Free Full Text].
|
| 7.
|
Fisher, N. L.,
L. W. Burgess,
T. A. Tourssoun, and P. E. Nelson.
1982.
Carnation leaves as substrate and for preserving cultures of Fusarium species.
Phytopathology
72:151-153.
|
| 8.
|
Guarro, J., and J. Gene.
1995.
Opportunistic fusarial infections in humans.
Eur. J. Clin. Microbiol. Infect. Dis.
14:741-754[Medline].
|
| 9.
|
Law, D.,
C. B. Moore, and D. W. Denning.
1997.
Activity of SCH 56592 compared with those of fluconazole and intraconazole against Candida spp.
Antimicrob. Agents Chemother.
41:2310-2311[Abstract].
|
| 10.
|
Lozano, M.,
J. M. Ribera,
J. Puig,
A. Rives,
J. Sierra,
R. Grañena, and C. Rozman.
1990.
Bronconeumonia por Fusarium solani en un paciente afectado de leucemia aguda mieloblastica.
Enferm. Infecc. Microbol. Clin.
8:78-79.
|
| 10a.
|
Lozano-Chiu, M.,
S. Arikan,
V. Paetznick,
E. J. Anaissle,
D. Loebenberg, and J. H. Rex.
1998.
Successful treatment of murine fusariosis with Schering (SCH) 56592, abstr. F-101, p. 270.
In
Abstracts of the 98th General Meeting of the American Society for Microbiology. American Society for Microbiology, Washington, D.C.
|
| 11.
|
Lutz, J. E.,
K. V. Clemons,
B. H. Aristizabal, and D. A. Stevens.
1997.
Activity of the triazole SCH 56592 against disseminated murine coccidioidomycosis.
Antimicrob. Agents Chemother.
41:1558-1561[Abstract].
|
| 12.
|
National Committee for Clinical Laboratory Standards.
1997.
Reference method for broth dilution antifungal susceptibility testing of yeasts. Approved standard M27-A.
National Committee for Clinical Laboratory Standards, Wayne, Pa.
|
| 13.
|
Nelson, P. E.,
M. C. Dignani, and E. J. Anaissie.
1994.
Taxonomy, biology, and clinical aspects of Fusarium species.
Clin. Microbiol. Rev.
7:479-504[Abstract/Free Full Text].
|
| 14.
|
Neumeister, B.,
P. Bartmann,
G. Gaedicke, and R. Marre.
1992.
A fatal infection due to Fusarium oxysporum in a child with Wilms's tumour. Case report and review of the literature.
Mycoses
35:115-119[Medline].
|
| 15.
|
Oakley, K. L.,
G. Morrisey, and D. W. Denning.
1997.
Efficacy of SCH 56592 in a temporarily neutropenic murine model of invasive aspergillosis with an itraconazole-susceptible and an itraconazole-resistant isolate of Aspergillus fumigatus.
Antimicrob. Agents Chemother.
41:1504-1507[Abstract].
|
| 16.
|
Pfaller, M. A.,
S. Messer, and R. N. Jones.
1997.
Activity of a new triazole, Sch 56592, compared with those of four antifungal agents tested against clinical isolates of Candida spp. and Saccharomyces cerevisiae.
Antimicrob. Agents Chemother.
41:233-235[Abstract].
|
| 17.
|
Pujol, I.,
J. Guarro,
J. Gene, and J. Sala.
1997.
In vitro antifungal susceptibility of clinical and environmental Fusarium spp. strains.
J. Antimicrob. Chemother.
39:163-167[Abstract/Free Full Text].
|
| 18.
|
Rabodonirina, M.,
M. A. Piens,
M. F. Monier,
E. Gueho,
D. Fiere, and M. Mojon.
1994.
Fusarium infections in immunocompromised patients: case reports and literature review.
Eur. J. Clin. Microbiol. Infect. Dis.
13:152-161[Medline].
|
| 19.
|
Reuben, A.,
E. Anaissie,
P. E. Nelson,
R. Hashem,
C. Legrand,
D. H. Ho, and G. P. Bodey.
1989.
Antifungal susceptibility of 44 clinical isolates of Fusarium species determined by using a broth microdilution method.
Antimicrob. Agents Chemother.
33:1647-1649[Abstract/Free Full Text].
|
| 20.
|
Sanche, S. E.,
D. A. Sutton,
K. Magnon,
P. Cox,
S. Revankar, and M. G. Rinaldi.
1998.
Psuedoepidemic of Fusarium oxysporum from bronchoscopy specimens, abstr. F-102, p. 270.
In
Abstracts of the 98th General Meeting of the American Society for Microbiology. American Society for Microbiology, Washington, D.C.
|
| 20a.
| Schering Plough Research Institute. Data on file.
|
| 21.
|
Sekhon, A. S.,
A. A. Padhye,
A. K. Garg,
H. Ahmad, and N. Moledina.
1994.
In vitro sensitivity of medically significant Fusarium species to various antimycotics.
Chemotherapy
40:239-244[Medline].
|
| 22.
|
Srdic, N.,
S. Radulovic,
Z. Nonkovic,
S. Velimirovic,
L. Cvetkovic, and I. Viro.
1993.
Two cases of exogenous endophthalmitis due to Fusarium moniliforme and Pseudomonas species as associated aetiological agents.
Mycoses
36:441-444[Medline].
|
| 23.
|
Sugar, A. M., and X.-P. Liu.
1996.
In vitro and in vivo activities of SCH 56592 against Blastomyces dermatitidis.
Antimicrob. Agents Chemother.
40:1314-1316[Abstract].
|
| 24.
|
Vartivarian, S. E.,
E. J. Anaissie, and G. P. Bodey.
1993.
Emerging fungal pathogens in immunocompromised patients: classification, diagnosis, and management.
Clin. Infect. Dis.
17:S487-S491.
|
Antimicrobial Agents and Chemotherapy, March 1999, p. 589-591, Vol. 43, No. 3
0066-4804/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
This article has been cited by other articles:
-
Marine, M., Pastor, F. J., Serena, C., Guarro, J.
(2009). Efficacy of Triazoles in a Murine Disseminated Infection by Candida krusei. Antimicrob. Agents Chemother.
53: 3585-3588
[Abstract]
[Full Text]
-
Rodriguez, M. M., Pastor, F. J., Serena, C., Guarro, J.
(2009). Posaconazole efficacy in a murine disseminated infection caused by Paecilomyces lilacinus. J Antimicrob Chemother
63: 361-364
[Abstract]
[Full Text]
-
Carrillo-Munoz, A.-J., Quindos, G., Ruesga, M., Alonso, R., del Valle, O., Hernandez-Molina, J. M., McNicholas, P., Loebenberg, D., Santos, P.
(2005). Antifungal activity of posaconazole compared with fluconazole and amphotericin B against yeasts from oropharyngeal candidiasis and other infections. J Antimicrob Chemother
55: 317-319
[Abstract]
[Full Text]
-
Simitsopoulou, M., Gil-Lamaignere, C., Avramidis, N., Maloukou, A., Lekkas, S., Havlova, E., Kourounaki, L., Loebenberg, D., Roilides, E.
(2004). Antifungal Activities of Posaconazole and Granulocyte-Macrophage Colony-Stimulating Factor Ex Vivo and in Mice with Disseminated Infection Due to Scedosporium prolificans. Antimicrob. Agents Chemother.
48: 3801-3805
[Abstract]
[Full Text]
-
Paphitou, N. I., Ostrosky-Zeichner, L., Paetznick, V. L., Rodriguez, J. R., Chen, E., Rex, J. H.
(2002). In Vitro Activities of Investigational Triazoles against Fusarium Species: Effects of Inoculum Size and Incubation Time on Broth Microdilution Susceptibility Test Results. Antimicrob. Agents Chemother.
46: 3298-3300
[Abstract]
[Full Text]
-
Barchiesi, F., Schimizzi, A. M., Najvar, L. K., Bocanegra, R., Caselli, F., Di Cesare, S., Giannini, D., Di Francesco, L. F., Giacometti, A., Carle, F., Scalise, G., Graybill, J. R.
(2001). Interactions of Posaconazole and Flucytosine against Cryptococcus neoformans. Antimicrob. Agents Chemother.
45: 1355-1359
[Abstract]
[Full Text]
-
Petraitiene, R., Petraitis, V., Groll, A. H., Sein, T., Piscitelli, S., Candelario, M., Field-Ridley, A., Avila, N., Bacher, J., Walsh, T. J.
(2001). Antifungal Activity and Pharmacokinetics of Posaconazole (SCH 56592) in Treatment and Prevention of Experimental Invasive Pulmonary Aspergillosis: Correlation with Galactomannan Antigenemia. Antimicrob. Agents Chemother.
45: 857-869
[Abstract]
[Full Text]
-
Cacciapuoti, A., Loebenberg, D., Corcoran, E., Menzel, F. Jr., Moss, E. L. Jr., Norris, C., Michalski, M., Raynor, K., Halpern, J., Mendrick, C., Arnold, B., Antonacci, B., Parmegiani, R., Yarosh-Tomaine, T., Miller, G. H., Hare, R. S.
(2000). In Vitro and In Vivo Activities of SCH 56592 (Posaconazole), a New Triazole Antifungal Agent, against Aspergillus and Candida. Antimicrob. Agents Chemother.
44: 2017-2022
[Abstract]
[Full Text]
-
Molina, J., Martins-Filho, O., Brener, Z., Romanha, A. J., Loebenberg, D., Urbina, J. A.
(2000). Activities of the Triazole Derivative SCH 56592 (Posaconazole) against Drug-Resistant Strains of the Protozoan Parasite Trypanosoma (Schizotrypanum) cruzi in Immunocompetent and Immunosuppressed Murine Hosts. Antimicrob. Agents Chemother.
44: 150-155
[Abstract]
[Full Text]
Top
Abstract
Introduction
