scholarly journals APX001 Is Effective in the Treatment of Murine Invasive Pulmonary Aspergillosis

2018 ◽  
Vol 63 (2) ◽  
pp. e01713-18 ◽  
Author(s):  
Teclegiorgis Gebremariam ◽  
Sondus Alkhazraji ◽  
Abdullah Alqarihi ◽  
Heewon H. Jeon ◽  
Yiyou Gu ◽  
...  
Keyword(s):  
Fungal Infections ◽  
Histopathological Examination ◽  
Invasive Pulmonary Aspergillosis ◽  
Human Pharmacokinetics ◽  
Immunosuppressed Patient ◽  
Cytochrome P450 Enzymes ◽  
Pulmonary Aspergillosis ◽  
Minimum Effective Concentration ◽  
Content Type

ABSTRACTInvasive pulmonary aspergillosis (IPA) due toAspergillus fumigatusis a serious fungal infection in the immunosuppressed patient population. Despite the introduction of new antifungal agents, mortality rates remain high, and new treatments are needed. The novel antifungal APX001A targets the conserved Gwt1 enzyme required for the localization of glycosylphosphatidylinositol-anchored mannoproteins in fungi. We evaluated thein vitroactivity of APX001A againstA. fumigatusand thein vivoactivity of its prodrug APX001 in an immunosuppressed mouse model of IPA. APX001A inhibited the growth ofA. fumigatuswith a minimum effective concentration of 0.03 μg/ml. The use of 50 mg/kg 1-aminobenzotriazole (ABT), a suicide inhibitor of cytochrome P450 enzymes, enhanced APX001A exposures (area under the time-concentration curve [AUC]) 16- to 18-fold and enhanced serum half-life from ∼1 to 9 h, more closely mimicking human pharmacokinetics. We evaluated the efficacy of APX001 (with ABT) in treating murine IPA compared to posaconazole treatment. Treatment of mice with 78 mg/kg once daily (QD), 78 mg/kg twice daily, or 104 mg/kg QD APX001 significantly enhanced the median survival time and prolonged day 21 postinfection overall survival compared to the placebo. Furthermore, administration of APX001 resulted in a significant reduction in lung fungal burden (4.2 to 7.6 log10conidial equivalents/g of tissue) versus the untreated control and resolved the infection, as judged by histopathological examination. The observed survival and tissue clearance were comparable to a clinically relevant posaconazole dose. These results warrant the continued development of APX001 as a broad-spectrum, first-in-class treatment of invasive fungal infections.

scholarly journals APX001 Pharmacokinetic/Pharmacodynamic Target Determination againstAspergillus fumigatusin anIn VivoModel of Invasive Pulmonary Aspergillosis

2019 ◽  
Vol 63 (4) ◽  
Author(s):  
Miao Zhao ◽  
Alexander J. Lepak ◽  
Karen Marchillo ◽  
Jamie Vanhecker ◽  
Hiram Sanchez ◽  
...  
Keyword(s):  
Invasive Pulmonary Aspergillosis ◽  
Dose Fractionation ◽  
Coefficient Of Determination ◽  
Dose Selection ◽  
Pulmonary Aspergillosis ◽  
Time Curve ◽  
Minimum Effective Concentration ◽  
Content Type

ABSTRACTAPX001, the prodrug of APX001A, is a first-in-class antifungal agent that has a potent activity againstAspergillus fumigatus. The goal of current study was to determine the pharmacodynamic (PD) index and target of APX001 in an immunocompromised murine model of invasive pulmonary aspergillosis against 6 A. fumigatusisolates. Minimum effective concentration (MEC) values ranged from 0.03 to 0.06 mg/liter. Dose fractionation was performed against isolate AF293 using total doses of APX001 ranging from 81 to 768 mg/kg of body weight/day fractionated into every 3-, 6-, and 8-h regimens over a 96-h treatment duration. Efficacy was assessed byA. fumigatusquantitative PCR (qPCR) of conidial equivalents from lung homogenates. Nonlinear regression analysis using the Hill equation demonstrated that the 24-h area under the concentration-time curve (AUC)/MEC ratio was the pharmacokinetic (PK)/PD index that best correlated with efficacy (coefficient of determination [R2] = 0.79). Treatment studies with the remaining strains utilized regimens of 40 to 1,536 mg/kg of APX001 administered every 3 h for a 96-h duration. Exposure-response relationships for all strains were similar, and the median free drug AUC/MEC PK/PD targets for stasis and 1-log-kill endpoints were 47.6 and 89.4, respectively. The present studies demonstratedin vitroandin vivoAPX001A/APX001 potency againstA. fumigatus. These results have potential relevance for clinical dose selection and evaluation of susceptibility breakpoints.

scholarly journals Antifungal effect of all-trans retinoic acid against Aspergillus fumigatus in vitro and in a pulmonary aspergillosis in vivo model

2020 ◽  
Author(s):  
Elena Campione ◽  
Roberta Gaziano ◽  
Elena Doldo ◽  
Daniele Marino ◽  
Mattia Falconi ◽  
...  
Keyword(s):  
Retinoic Acid ◽  
Aspergillus Fumigatus ◽  
Rat Model ◽  
Fungal Infections ◽  
Invasive Pulmonary Aspergillosis ◽  
Antifungal Drugs ◽  
Pulmonary Aspergillosis ◽  
Fungistatic Activity

AIM: Aspergillus fumigatus is the most common opportunistic fungal pathogen and causes invasive pulmonary aspergillosis (IPA), with high mortality among immunosuppressed patients. Fungistatic activity of all-trans retinoic acid (ATRA) has been recently described in vitro. We evaluated the efficacy of ATRA in vivo and its potential synergistic interaction with other antifungal drugs. MATERIALS AND METHODS: A rat model of IPA and in vitro experiments were performed to assess the efficacy of ATRA against Aspergillus in association with classical antifungal drugs and in silico studies used to clarify its mechanism of action. RESULTS: ATRA (0.5 and 1 mM) displayed a strong fungistatic activity in Aspergillus cultures, while at lower concentrations, synergistically potentiated fungistatic efficacy of sub-inhibitory concentration of Amphotericin B (AmB) and Posaconazole (POS). ATRA also enhanced macrophagic phagocytosis of conidia. In a rat model of IPA, ATRA reduced mortality similarly to Posaconazole. CONCLUSION: Fungistatic efficacy of ATRA alone and synergistically with other antifungal drugs was documented in vitro, likely by inhibiting fungal Hsp90 expression and Hsp90-related genes. ATRA reduced mortality in a model of IPA in vivo. Those findings suggest ATRA as suitable fungistatic agent, also to reduce dosage and adverse reaction of classical antifungal drugs, and new therapeutic strategies against IPA and systemic fungal infections.

scholarly journals Chitinase Induction Prior to Caspofungin Treatment of Experimental Invasive Aspergillosis in Neutropenic Rats Does Not Enhance Survival

2017 ◽  
Vol 62 (1) ◽  
Author(s):  
Jeannine M. Refos ◽  
Alieke G. Vonk ◽  
Marian T. ten Kate ◽  
Henri A. Verbrugh ◽  
Irma A. J. M. Bakker-Woudenberg ◽  
...  
Keyword(s):  
Antifungal Activity ◽  
Invasive Aspergillosis ◽  
Invasive Pulmonary Aspergillosis ◽  
Left Lung ◽  
Chitinase Activity ◽  
Pulmonary Aspergillosis ◽  
Content Type ◽  
Acidic Mammalian Chitinase ◽  
Phosphate Buffered Saline

ABSTRACT Host chitinases, chitotriosidase and acidic mammalian chitinase (AMCase), improved the antifungal activity of caspofungin (CAS) against Aspergillus fumigatus in vitro. These chitinases are not constitutively expressed in the lung. Here, we investigated whether chitosan derivatives were able to induce chitinase activity in the lungs of neutropenic rats and, if so, whether these chitinases were able to prolong survival of rats with invasive pulmonary aspergillosis (IPA) or of rats with IPA and treated with CAS. An oligosaccharide-lactate chitosan (OLC) derivative was instilled in the left lung of neutropenic rats to induce chitotriosidase and AMCase activities. Rats instilled with OLC or with phosphate-buffered saline (PBS) were subsequently infected with A. fumigatus and then treated with suboptimal doses of CAS. Survival, histopathology, and galactomannan indexes were determined. Instillation of OLC resulted in chitotriosidase and AMCase activities. However, instillation of OLC did not prolong rat survival when rats were subsequently challenged with A. fumigatus. In 5 of 7 rats instilled with OLC, the fungal foci in the lungs were smaller than those in rats instilled with PBS. Instillation of OLC did not significantly enhance the survival of neutropenic rats challenged with A. fumigatus and treated with a suboptimal dosage of CAS. Chitotriosidase and AMCase activities can be induced with OLC, but the presence of active chitinases in the lung did not prevent the development of IPA or significantly enhance the therapeutic outcome of CAS treatment.

ADAMTS13 Regulates Neutrophil Recruitment in a Mouse Model of Invasive Pulmonary Aspergillosis

Blood ◽  
2014 ◽  
Vol 124 (21) ◽  
pp. 4109-4109
Author(s):  
Markus Radsak ◽  
Steve Prüfer ◽  
Katharina Ebner ◽  
Michael Weber ◽  
Sebastian Reuter ◽  
...  
Keyword(s):  
Inflammatory Response ◽  
Mouse Model ◽  
Aspergillus Fumigatus ◽  
Fungal Infections ◽  
Invasive Pulmonary Aspergillosis ◽  
Polymorphonuclear Neutrophils ◽  
Formyl Peptide Receptor ◽  
Wild Type ◽  
Pulmonary Aspergillosis

Abstract Von Willebrand factor (VWF) is secreted as an acute phase protein during inflammation. The main mechanism regulating the size and prothrombotic activity of VWF is the specific proteolytic activity of ADAMTS-13. To determine the relevance of this regulatory pathway for the innate inflammatory response by polymorphonuclear neutrophils (PMN), we employed a mouse model of invasive pulmonary aspergillosis (IPA) where PMN functionality is crucial for fungal clearance and survival. IPA was induced by intratracheal application of Aspergillus fumigatus conidia in wild-type (129/Sv/Pas) or Adamts13 deficient (Adamts13-/-) mice. After PMN depletion using a anti-Gr-1 specific antibody, all mice infected with Aspergillus fumigatus conidia developed neutropenia and succumbed due to lethal IPA. In contrast, all undepleted wild-type mice survived the infection. Interestingly, Aspergillus fumigatus infection in Adamts13-/- mice was lethal in 20% of the animals displaying a more severe course of IPA, as indicated by an increased fungal burden in lung homogenates along with increased levels of albumin and the inflammatory mediators IL-1β, IL-6, TNF-α, KC and MCP-1 in the bronchio-alveolar lavage fluid (BALF) compared to wild-type controls. Beyond this, we observed a decreased number of PMN in BALF of infected Adamts13-/- mice compared to wild-type mice. Lung histology sections demonstrated a more pronounced perivascular leukocyte infiltration in further support of a dysregulated inflammatory response in Adamts13-/- mice. Importantly, we observed no general defect in the activation of neutrophil effector functions as demonstrated by the normal induction of the oxidative burst, phagocytosis, degranulation, L-selectin shedding and apoptosis in response to formyl-peptide receptor agonists or exposure to Aspergillus fumigatus conidia or hyphae in vitro. Therefore, we conclude that the proteolytic regulation of VWF by ADAMTS-13 in an important mechanism to control PMN recruitment in the regulation of the innate inflammatory response in invasive fungal infections. Disclosures Radsak: Celgene: Research Funding.

scholarly journals Host Biomarkers of Invasive Pulmonary Aspergillosis To Monitor Therapeutic Response

2014 ◽  
Vol 58 (6) ◽  
pp. 3373-3378 ◽  
Author(s):  
Mila Krel ◽  
Vidmantas Petraitis ◽  
Ruta Petraitiene ◽  
Mohit Raja Jain ◽  
Yanan Zhao ◽  
...  
Keyword(s):  
Therapeutic Response ◽  
Invasive Pulmonary Aspergillosis ◽  
Rabbit Model ◽  
Time Of Flight ◽  
Lavage Fluid ◽  
Host Protein ◽  
Enzyme Linked Immunosorbent Assay ◽  
Pulmonary Aspergillosis ◽  
Content Type

ABSTRACTInvasive pulmonary aspergillosis (IPA) is a life-threatening disease of immunocompromised patients that requires aggressive therapy. Detection of the disease and monitoring of the therapeutic response during IPA are complex, and current molecular diagnostics are not suitably robust. Here, we explored proteomic profiles of bronchoalveolar lavage fluid (BALF) specimens from a persistently neutropenic rabbit model of IPA. Three experimental arms, uninfected control animals, infected untreated animals, and animals infected and treated with ravuconazole/amphotericin B, were studied. Total proteins were evaluated by two-dimensional (2D) gel electrophoresis, followed by matrix-assisted laser desorption ionization–time of flight/time of flight (MALDI-TOF/TOF) mass spectrometry (MS) and quantified by enzyme-linked immunosorbent assay (ELISA). Host-derived proteins haptoglobin (Hp), C-reactive protein (CRP), and annexin A1 (Anx A1) were prominently found in BALF during the IPA infection and showed significant changes in response to antifungal therapy (P< 0.0001). In serum, differences in Hp (P= 0.0001) between infected and treated rabbits were observed. Preliminaryin vitrostudies revealed thatAspergillus fumigatus-secreted proteases may contribute to the cleavage of Anx A1 during IPA. In summary, host protein biomarkers Hp, CRP, and Anx A1 may have value in monitoring therapeutic response to antifungal agents in IPA patients with confirmed disease.

scholarly journals A Novel Polyaminocarboxylate Compound To Treat Murine Pulmonary Aspergillosis by Interfering with Zinc Metabolism

2018 ◽  
Vol 62 (6) ◽  
Author(s):  
Paris Laskaris ◽  
Rocío Vicentefranqueira ◽  
Olivier Helynck ◽  
Grégory Jouvion ◽  
José Antonio Calera ◽  
...  
Keyword(s):  
Fungal Infections ◽  
Treatment Options ◽  
Fungal Growth ◽  
Opportunistic Pathogen ◽  
Zinc Metabolism ◽  
Pulmonary Aspergillosis ◽  
Hyphal Length ◽  
Content Type

ABSTRACT Aspergillus fumigatus can cause pulmonary aspergillosis in immunocompromised patients and is associated with a high mortality rate due to a lack of reliable treatment options. This opportunistic pathogen requires zinc in order to grow and cause disease. Novel compounds that interfere with fungal zinc metabolism may therefore be of therapeutic interest. We screened chemical libraries containing 59,223 small molecules using a resazurin assay that compared their effects on an A. fumigatus wild-type strain grown under zinc-limiting conditions and on a zinc transporter knockout strain grown under zinc-replete conditions to identify compounds affecting zinc metabolism. After a first screen, 116 molecules were selected whose inhibitory effects on fungal growth were further tested by using luminescence assays and hyphal length measurements to confirm their activity, as well as by toxicity assays on HeLa cells and mice. Six compounds were selected following a rescreening, of which two were pyrazolones, two were porphyrins, and two were polyaminocarboxylates. All three groups showed good in vitro activity, but only one of the polyaminocarboxylates was able to significantly improve the survival of immunosuppressed mice suffering from pulmonary aspergillosis. This two-tier screening approach led us to the identification of a novel small molecule with in vivo fungicidal effects and low murine toxicity that may lead to the development of new treatment options for fungal infections by administration of this compound either as a monotherapy or as part of a combination therapy.

scholarly journals Fosmanogepix (APX001) Is Effective in the Treatment of Pulmonary Murine Mucormycosis Due to Rhizopus arrhizus

2020 ◽  
Vol 64 (6) ◽  
Author(s):  
Teclegiorgis Gebremariam ◽  
Sondus Alkhazraji ◽  
Abdullah Alqarihi ◽  
Nathan P. Wiederhold ◽  
Karen Joy Shaw ◽  
...  
Keyword(s):  
Fungal Infections ◽  
Surface Proteins ◽  
Rhizopus Arrhizus ◽  
Minimum Effective Concentration ◽  
Content Type ◽  
Log Reduction ◽  
Infection Models ◽  
Life Threatening

ABSTRACT Mucormycosis is a life-threatening infection with high mortality that occurs predominantly in immunocompromised patients. Manogepix (MGX) is a novel antifungal that targets Gwt1, a protein involved in an early step in the conserved glycosylphosphotidyl inositol (GPI) posttranslational modification pathway of surface proteins in eukaryotic cells. Inhibition of fungal inositol acylation by MGX results in pleiotropic effects, including inhibition of maturation of GPI-anchored proteins necessary for growth and virulence. MGX has been previously shown to have in vitro activity against some strains of Mucorales. Here, we assessed the in vivo activity of the prodrug fosmanogepix, currently in clinical development for the treatment of invasive fungal infections, against two Rhizopus arrhizus strains with high (4.0 μg/ml) and low (0.25 μg/ml) minimum effective concentration (MEC) values. In both invasive pulmonary infection models, treatment of mice with 78 mg/kg or 104 mg/kg fosmanogepix, along with 1-aminobenzotriazole to enhance the serum half-life of MGX in mice, significantly increased median survival time and prolonged overall survival by day 21 postinfection compared to placebo. In addition, administration of fosmanogepix resulted in a 1 to 2 log reduction in both lung and brain fungal burden. For the 104 mg/kg fosmanogepix dose, tissue clearance and survival were comparable to clinically relevant doses of isavuconazole (ISA), which is FDA approved for the treatment of mucormycosis. These results support continued development of fosmanogepix as a first-in-class treatment for invasive mucormycosis.

scholarly journals Hyperbaric Oxygen ReducesAspergillus fumigatusProliferationIn Vitroand InfluencesIn VivoDisease Outcomes

2017 ◽  
Vol 62 (3) ◽  
Author(s):  
Sourabh Dhingra ◽  
Jay C. Buckey ◽  
Robert A. Cramer
Keyword(s):  
Hyperbaric Oxygen ◽  
Fungal Infections ◽  
Invasive Pulmonary Aspergillosis ◽  
Modest Improvement ◽  
Content Type ◽  
Disease Outcomes ◽  
Treatment Conditions ◽  
Dose Dependent

ABSTRACTRecent estimates suggest that more than 3 million people have chronic or invasive fungal infections, causing more than 600,000 deaths every year.Aspergillus fumigatuscauses invasive pulmonary aspergillosis (IPA) in patients with compromised immune systems and is a primary contributor to increases in human fungal infections. Thus, the development of new clinical modalities as stand-alone or adjunctive therapy for improving IPA patient outcomes is critically needed. Here we tested thein vitroandin vivoimpacts of hyperbaric oxygen (HBO) (100% oxygen, >1 atmosphere absolute [ATA]) onA. fumigatusproliferation and murine IPA outcomes. Our findings indicate that HBO reduces established fungal biofilm proliferationin vitroby over 50%. The effect of HBO under the treatment conditions was transient and fungistatic, withA. fumigatusmetabolic activity rebounding within 6 h of HBO treatment being removed.In vivo, daily HBO provides a dose-dependent but modest improvement in murine IPA disease outcomes as measured by survival analysis. Intriguingly, no synergy was observed between subtherapeutic voriconazole or amphotericin B and HBOin vitroorin vivowith daily HBO dosing, though the loss of fungal superoxide dismutase genes enhanced HBO antifungal activity. Further studies are needed to optimize the HBO treatment regimen and better understand the effects of HBO on both the host and the pathogen during a pulmonary invasive fungal infection.

scholarly journals Exploration of Sulfur Assimilation of Aspergillus fumigatus Reveals Biosynthesis of Sulfur-Containing Amino Acids as a Virulence Determinant

2016 ◽  
Vol 84 (4) ◽  
pp. 917-929 ◽  
Author(s):  
Jorge Amich ◽  
Michaela Dümig ◽  
Gráinne O'Keeffe ◽  
Jasmin Binder ◽  
Sean Doyle ◽  
...  
Keyword(s):  
Amino Acid ◽  
Aspergillus Fumigatus ◽  
Mutant Strain ◽  
Fungal Infections ◽  
Invasive Pulmonary Aspergillosis ◽  
Delayed Diagnosis ◽  
Phenotypic Characterization ◽  
Nutritional Requirements ◽  
Pulmonary Aspergillosis ◽  
Content Type

Fungal infections are of major relevance due to the increased numbers of immunocompromised patients, frequently delayed diagnosis, and limited therapeutics. To date, the growth and nutritional requirements of fungi during infection, which are relevant for invasion of the host, are poorly understood. This is particularly true for invasive pulmonary aspergillosis, as so far, sources of (macro)elements that are exploited during infection have been identified to only a limited extent. Here, we have investigated sulfur (S) utilization by the human-pathogenic moldAspergillus fumigatusduring invasive growth. Our data reveal that inorganic S compounds or taurine is unlikely to serve as an S source during invasive pulmonary aspergillosis since a sulfate transporter mutant strain and a sulfite reductase mutant strain are fully virulent. In contrast, the S-containing amino acid cysteine is limiting for fungal growth, as proven by the reduced virulence of a cysteine auxotroph. Moreover, phenotypic characterization of this strain further revealed the robustness of the subordinate glutathione redox system. Interestingly, we demonstrate that methionine synthase is essential forA. fumigatusvirulence, defining the biosynthetic route of this proteinogenic amino acid as a potential antifungal target. In conclusion, we provide novel insights into the nutritional requirements ofA. fumigatusduring pathogenesis, a prerequisite to understanding and fighting infection.

scholarly journals Combination of Voriconazole and Anidulafungin for Treatment of Triazole-Resistant Aspergillus fumigatus in anIn VitroModel of Invasive Pulmonary Aspergillosis

2012 ◽  
Vol 56 (10) ◽  
pp. 5180-5185 ◽  
Author(s):  
Adam R. Jeans ◽  
Susan J. Howard ◽  
Zaid Al-Nakeeb ◽  
Joanne Goodwin ◽  
Lea Gregson ◽  
...  
Keyword(s):  
High Performance ◽  
Molecular Mechanisms ◽  
Invasive Pulmonary Aspergillosis ◽  
Fungal Growth ◽  
Pulmonary Aspergillosis ◽  
Antifungal Effect ◽  
Content Type ◽  
Exposure Response ◽  
Relationship Of

ABSTRACTVoriconazole is a first-line agent for the treatment of invasive pulmonary aspergillosis. Isolates with elevated voriconazole MICs are increasingly being seen, and the optimal treatment regimen is not defined. We investigated whether the combination of voriconazole with anidulafungin may be beneficial for the treatment ofA. fumigatusstrains with elevated voriconazole MICs. We used anin vitromodel of the human alveolus to define the exposure-response relationships for a wild-type strain (voriconazole MIC, 0.5 mg/liter) and strains with defined molecular mechanisms of triazole resistance (MICs, 4 to 16 mg/liter). All strains had anidulafungin minimum effective concentrations (MECs) of 0.0078 mg/liter. Exposure-response relationships were estimated using galactomannan as a biomarker. Concentrations of voriconazole and anidulafungin were measured using high-performance liquid chromatography (HPLC). The interaction of voriconazole and anidulafungin was described using the Greco model. Fungal growth was progressively inhibited with higher drug exposures of voriconazole. Strains with elevated voriconazole MICs required proportionally greater voriconazole exposures to achieve a comparable antifungal effect. Galactomannan concentrations were only marginally reduced by anidulafungin monotherapy. An additive effect between voriconazole and anidulafungin was apparent. In conclusion, the addition of anidulafungin does not markedly alter the exposure-response relationship of voriconazole. A rise in serum galactomannan during combination therapy with voriconazole and anidulafungin should be interpreted as treatment failure and not attributed to a paradoxical reaction related to echinocandin treatment.

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