Peptide Information
General Information of This Peptide
| Peptide ID |
PEP01069
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| Peptide Name |
HLopt2-NH<sub>2</sub>
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| Sequence |
CFQWKRAMRKVR
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| Peptide Type |
Linear
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| PDC Transmembrane Types | Cell targeting peptides (CTPs) | |||||
Each Peptide-drug Conjugate Related to This Peptide
Full Information of The Activity Data of The PDC(s) Related to This Peptide
FLCpOH-HLopt2-NH2 [Investigative]
Revealed Based on the Cell Line Data
| Experiment 1 Reporting the Activity Data of This PDC | [1] | ||||
| Indication | <i>Candida spp</i> infection | ||||
| Efficacy Data | Minimum fungicidal concentration |
62 μM
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| Administration Time | 24 h | ||||
| Evaluation Method | Antifungal activity of the tested compounds against yeast cells was determined in 96-well plates, with minor modifications (CLSI, 2008). Suspensions of the microorganisms were prepared by taking one loop of pure culture into sterile water and adjusting optical density to 0.1 at 660 nm wavelength and further 50-fold dilution in RPMI 1640 medium, resulting in cell concentration of approximately 2 104 CFU/m. Then, 100 μL of cells were added to the wells of a 96-well microtiter plate that contained 16-500 μM tested compounds and 16-500 μM of FLC or 0.2-62 μM AmB. The plates were incubated for 24 h at 37 °C. Each test was performed in triplicate. The final concentration of DMSO was ensured to be around 1% in all experiments. The MIC90 was defined as the lowest concentration of drug showing no visible growth; MIC50 was defined as the concentration that yielded at least 50% growth inhibition when compared with the growth control well. For determination of minimum fungicidal concentrations (MFC), small aliquots of suspensions (around 10 μL) from each well were transferred using the pipette to YPD agar plates without inhibitors and incubated for 24 h at 37 °C. The MFC was defined as the lowest concentration of drug at which no growth of the colonies was observed. | ||||
| MOA of PDC |
Herein we report the chemical synthesis and biological activity of conjugates of fluconazole with (i) cell-penetrating peptides (CPP), namely TP10-NH2 and TP10-7-NH2, or (ii) antimicrobial peptides (AMP), such as LFcinB(2-11)-NH2, LFcinB[Nle1,11]-NH2 and HLopt2-NH2. Both constituents of produced conjugates (FLC and peptides) display different modes of antifungal activity and affect different molecular targets within fungal cells. CPPs serving as carrier peptides and are considered the fundamental part of an extensively developed concept of a drug delivery system; TP-10 is one of the most promising peptides in this family. These peptides effectively penetrate cell membrane. In our recent paper. we showed that TP10-NH2 can be successfully applied to design conjugates with antimicrobial activity. Modified fragments of bovine lactoferrin (LFcin) and human lactoferrin (HLopt2) are members of the AMP family, peptides that constitute an important segment of the bodys natural immunity against microorganisms.
Click to Show/Hide
|
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| Description |
The inactivity of FLCpOH confirmed that the drug derivative, used during the synthesis of our target compounds, did not exert any antifungal activity. All compounds displayed poor activity against the C. glabrata strain tested. Another identified advantage of FLCpOH-TP10-NH2 and FLCpOH-TP10-7-NH2 is their higher fungicidal potential compared to fluconazole. The MFC values against C. albicans reference strains were the same or only twice higher compared to MIC90. The high fungicidal activity of both conjugates was also confirmed with the kill-time assay. However, at least a 4 x MFC concentration was necessary to obtain an evident killing effect after 24 h of treatment.
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| In Vitro Model | Staphylococcus aureus infection | Staphylococcus aureus | 1280 | ||
| Experiment 2 Reporting the Activity Data of This PDC | [1] | ||||
| Indication | <i>Candida spp</i> infection | ||||
| Efficacy Data | Minimum fungicidal concentration |
125 μM
|
|||
| Administration Time | 24 h | ||||
| Evaluation Method | Antifungal activity of the tested compounds against yeast cells was determined in 96-well plates, with minor modifications (CLSI, 2008). Suspensions of the microorganisms were prepared by taking one loop of pure culture into sterile water and adjusting optical density to 0.1 at 660 nm wavelength and further 50-fold dilution in RPMI 1640 medium, resulting in cell concentration of approximately 2 104 CFU/m. Then, 100 μL of cells were added to the wells of a 96-well microtiter plate that contained 16-500 μM tested compounds and 16-500 μM of FLC or 0.2-62 μM AmB. The plates were incubated for 24 h at 37 °C. Each test was performed in triplicate. The final concentration of DMSO was ensured to be around 1% in all experiments. The MIC90 was defined as the lowest concentration of drug showing no visible growth; MIC50 was defined as the concentration that yielded at least 50% growth inhibition when compared with the growth control well. For determination of minimum fungicidal concentrations (MFC), small aliquots of suspensions (around 10 μL) from each well were transferred using the pipette to YPD agar plates without inhibitors and incubated for 24 h at 37 °C. The MFC was defined as the lowest concentration of drug at which no growth of the colonies was observed. | ||||
| MOA of PDC |
Herein we report the chemical synthesis and biological activity of conjugates of fluconazole with (i) cell-penetrating peptides (CPP), namely TP10-NH2 and TP10-7-NH2, or (ii) antimicrobial peptides (AMP), such as LFcinB(2-11)-NH2, LFcinB[Nle1,11]-NH2 and HLopt2-NH2. Both constituents of produced conjugates (FLC and peptides) display different modes of antifungal activity and affect different molecular targets within fungal cells. CPPs serving as carrier peptides and are considered the fundamental part of an extensively developed concept of a drug delivery system; TP-10 is one of the most promising peptides in this family. These peptides effectively penetrate cell membrane. In our recent paper. we showed that TP10-NH2 can be successfully applied to design conjugates with antimicrobial activity. Modified fragments of bovine lactoferrin (LFcin) and human lactoferrin (HLopt2) are members of the AMP family, peptides that constitute an important segment of the bodys natural immunity against microorganisms.
Click to Show/Hide
|
||||
| Description |
The inactivity of FLCpOH confirmed that the drug derivative, used during the synthesis of our target compounds, did not exert any antifungal activity. All compounds displayed poor activity against the C. glabrata strain tested. Another identified advantage of FLCpOH-TP10-NH2 and FLCpOH-TP10-7-NH2 is their higher fungicidal potential compared to fluconazole. The MFC values against C. albicans reference strains were the same or only twice higher compared to MIC90. The high fungicidal activity of both conjugates was also confirmed with the kill-time assay. However, at least a 4 x MFC concentration was necessary to obtain an evident killing effect after 24 h of treatment.
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| In Vitro Model | Hepatocellular carcinoma | MHCC97H cell | CVCL_4972 | ||
| Experiment 3 Reporting the Activity Data of This PDC | [1] | ||||
| Indication | <i>Candida spp</i> infection | ||||
| Efficacy Data | Minimum fungicidal concentration | > 250 μM | |||
| Administration Time | 24 h | ||||
| Evaluation Method | Antifungal activity of the tested compounds against yeast cells was determined in 96-well plates, with minor modifications (CLSI, 2008). Suspensions of the microorganisms were prepared by taking one loop of pure culture into sterile water and adjusting optical density to 0.1 at 660 nm wavelength and further 50-fold dilution in RPMI 1640 medium, resulting in cell concentration of approximately 2 104 CFU/m. Then, 100 μL of cells were added to the wells of a 96-well microtiter plate that contained 16-500 μM tested compounds and 16-500 μM of FLC or 0.2-62 μM AmB. The plates were incubated for 24 h at 37 °C. Each test was performed in triplicate. The final concentration of DMSO was ensured to be around 1% in all experiments. The MIC90 was defined as the lowest concentration of drug showing no visible growth; MIC50 was defined as the concentration that yielded at least 50% growth inhibition when compared with the growth control well. For determination of minimum fungicidal concentrations (MFC), small aliquots of suspensions (around 10 μL) from each well were transferred using the pipette to YPD agar plates without inhibitors and incubated for 24 h at 37 °C. The MFC was defined as the lowest concentration of drug at which no growth of the colonies was observed. | ||||
| MOA of PDC |
Herein we report the chemical synthesis and biological activity of conjugates of fluconazole with (i) cell-penetrating peptides (CPP), namely TP10-NH2 and TP10-7-NH2, or (ii) antimicrobial peptides (AMP), such as LFcinB(2-11)-NH2, LFcinB[Nle1,11]-NH2 and HLopt2-NH2. Both constituents of produced conjugates (FLC and peptides) display different modes of antifungal activity and affect different molecular targets within fungal cells. CPPs serving as carrier peptides and are considered the fundamental part of an extensively developed concept of a drug delivery system; TP-10 is one of the most promising peptides in this family. These peptides effectively penetrate cell membrane. In our recent paper. we showed that TP10-NH2 can be successfully applied to design conjugates with antimicrobial activity. Modified fragments of bovine lactoferrin (LFcin) and human lactoferrin (HLopt2) are members of the AMP family, peptides that constitute an important segment of the bodys natural immunity against microorganisms.
Click to Show/Hide
|
||||
| Description |
The inactivity of FLCpOH confirmed that the drug derivative, used during the synthesis of our target compounds, did not exert any antifungal activity. All compounds displayed poor activity against the C. glabrata strain tested. Another identified advantage of FLCpOH-TP10-NH2 and FLCpOH-TP10-7-NH2 is their higher fungicidal potential compared to fluconazole. The MFC values against C. albicans reference strains were the same or only twice higher compared to MIC90. The high fungicidal activity of both conjugates was also confirmed with the kill-time assay. However, at least a 4 x MFC concentration was necessary to obtain an evident killing effect after 24 h of treatment.
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|
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| In Vitro Model | Mouse colon adenocarcinoma | MC-38 cell | CVCL_B288 | ||
| Experiment 4 Reporting the Activity Data of This PDC | [1] | ||||
| Indication | <i>Candida spp</i> infection | ||||
| Efficacy Data | Minimum fungicidal concentration | > 250 μM | |||
| Administration Time | 24 h | ||||
| Evaluation Method | Antifungal activity of the tested compounds against yeast cells was determined in 96-well plates, with minor modifications (CLSI, 2008). Suspensions of the microorganisms were prepared by taking one loop of pure culture into sterile water and adjusting optical density to 0.1 at 660 nm wavelength and further 50-fold dilution in RPMI 1640 medium, resulting in cell concentration of approximately 2 104 CFU/m. Then, 100 μL of cells were added to the wells of a 96-well microtiter plate that contained 16-500 μM tested compounds and 16-500 μM of FLC or 0.2-62 μM AmB. The plates were incubated for 24 h at 37 °C. Each test was performed in triplicate. The final concentration of DMSO was ensured to be around 1% in all experiments. The MIC90 was defined as the lowest concentration of drug showing no visible growth; MIC50 was defined as the concentration that yielded at least 50% growth inhibition when compared with the growth control well. For determination of minimum fungicidal concentrations (MFC), small aliquots of suspensions (around 10 μL) from each well were transferred using the pipette to YPD agar plates without inhibitors and incubated for 24 h at 37 °C. The MFC was defined as the lowest concentration of drug at which no growth of the colonies was observed. | ||||
| MOA of PDC |
Herein we report the chemical synthesis and biological activity of conjugates of fluconazole with (i) cell-penetrating peptides (CPP), namely TP10-NH2 and TP10-7-NH2, or (ii) antimicrobial peptides (AMP), such as LFcinB(2-11)-NH2, LFcinB[Nle1,11]-NH2 and HLopt2-NH2. Both constituents of produced conjugates (FLC and peptides) display different modes of antifungal activity and affect different molecular targets within fungal cells. CPPs serving as carrier peptides and are considered the fundamental part of an extensively developed concept of a drug delivery system; TP-10 is one of the most promising peptides in this family. These peptides effectively penetrate cell membrane. In our recent paper. we showed that TP10-NH2 can be successfully applied to design conjugates with antimicrobial activity. Modified fragments of bovine lactoferrin (LFcin) and human lactoferrin (HLopt2) are members of the AMP family, peptides that constitute an important segment of the bodys natural immunity against microorganisms.
Click to Show/Hide
|
||||
| Description |
The inactivity of FLCpOH confirmed that the drug derivative, used during the synthesis of our target compounds, did not exert any antifungal activity. All compounds displayed poor activity against the C. glabrata strain tested. Another identified advantage of FLCpOH-TP10-NH2 and FLCpOH-TP10-7-NH2 is their higher fungicidal potential compared to fluconazole. The MFC values against C. albicans reference strains were the same or only twice higher compared to MIC90. The high fungicidal activity of both conjugates was also confirmed with the kill-time assay. However, at least a 4 x MFC concentration was necessary to obtain an evident killing effect after 24 h of treatment.
Click to Show/Hide
|
||||
| In Vitro Model | Klebsiella pneumoniae infection | Klebsiella pneumoniae | 573 | ||
| Experiment 5 Reporting the Activity Data of This PDC | [1] | ||||
| Indication | <i>Candida spp</i> infection | ||||
| Efficacy Data | Minimum fungicidal concentration | > 250 μM | |||
| Administration Time | 24 h | ||||
| Evaluation Method | Antifungal activity of the tested compounds against yeast cells was determined in 96-well plates, with minor modifications (CLSI, 2008). Suspensions of the microorganisms were prepared by taking one loop of pure culture into sterile water and adjusting optical density to 0.1 at 660 nm wavelength and further 50-fold dilution in RPMI 1640 medium, resulting in cell concentration of approximately 2 104 CFU/m. Then, 100 μL of cells were added to the wells of a 96-well microtiter plate that contained 16-500 μM tested compounds and 16-500 μM of FLC or 0.2-62 μM AmB. The plates were incubated for 24 h at 37 °C. Each test was performed in triplicate. The final concentration of DMSO was ensured to be around 1% in all experiments. The MIC90 was defined as the lowest concentration of drug showing no visible growth; MIC50 was defined as the concentration that yielded at least 50% growth inhibition when compared with the growth control well. For determination of minimum fungicidal concentrations (MFC), small aliquots of suspensions (around 10 μL) from each well were transferred using the pipette to YPD agar plates without inhibitors and incubated for 24 h at 37 °C. The MFC was defined as the lowest concentration of drug at which no growth of the colonies was observed. | ||||
| MOA of PDC |
Herein we report the chemical synthesis and biological activity of conjugates of fluconazole with (i) cell-penetrating peptides (CPP), namely TP10-NH2 and TP10-7-NH2, or (ii) antimicrobial peptides (AMP), such as LFcinB(2-11)-NH2, LFcinB[Nle1,11]-NH2 and HLopt2-NH2. Both constituents of produced conjugates (FLC and peptides) display different modes of antifungal activity and affect different molecular targets within fungal cells. CPPs serving as carrier peptides and are considered the fundamental part of an extensively developed concept of a drug delivery system; TP-10 is one of the most promising peptides in this family. These peptides effectively penetrate cell membrane. In our recent paper. we showed that TP10-NH2 can be successfully applied to design conjugates with antimicrobial activity. Modified fragments of bovine lactoferrin (LFcin) and human lactoferrin (HLopt2) are members of the AMP family, peptides that constitute an important segment of the bodys natural immunity against microorganisms.
Click to Show/Hide
|
||||
| Description |
The inactivity of FLCpOH confirmed that the drug derivative, used during the synthesis of our target compounds, did not exert any antifungal activity. All compounds displayed poor activity against the C. glabrata strain tested. Another identified advantage of FLCpOH-TP10-NH2 and FLCpOH-TP10-7-NH2 is their higher fungicidal potential compared to fluconazole. The MFC values against C. albicans reference strains were the same or only twice higher compared to MIC90. The high fungicidal activity of both conjugates was also confirmed with the kill-time assay. However, at least a 4 x MFC concentration was necessary to obtain an evident killing effect after 24 h of treatment.
Click to Show/Hide
|
||||
| In Vitro Model | Pancreatic ductal adenocarcinoma | MIA PaCa-2 (KRAS G12C) cell | CVCL_0428 | ||
| Experiment 6 Reporting the Activity Data of This PDC | [1] | ||||
| Indication | <i>Candida spp</i> infection | ||||
| Efficacy Data | Minimum fungicidal concentration | > 250 μM | |||
| Administration Time | 24 h | ||||
| Evaluation Method | Antifungal activity of the tested compounds against yeast cells was determined in 96-well plates, with minor modifications (CLSI, 2008). Suspensions of the microorganisms were prepared by taking one loop of pure culture into sterile water and adjusting optical density to 0.1 at 660 nm wavelength and further 50-fold dilution in RPMI 1640 medium, resulting in cell concentration of approximately 2 104 CFU/m. Then, 100 μL of cells were added to the wells of a 96-well microtiter plate that contained 16-500 μM tested compounds and 16-500 μM of FLC or 0.2-62 μM AmB. The plates were incubated for 24 h at 37 °C. Each test was performed in triplicate. The final concentration of DMSO was ensured to be around 1% in all experiments. The MIC90 was defined as the lowest concentration of drug showing no visible growth; MIC50 was defined as the concentration that yielded at least 50% growth inhibition when compared with the growth control well. For determination of minimum fungicidal concentrations (MFC), small aliquots of suspensions (around 10 μL) from each well were transferred using the pipette to YPD agar plates without inhibitors and incubated for 24 h at 37 °C. The MFC was defined as the lowest concentration of drug at which no growth of the colonies was observed. | ||||
| MOA of PDC |
Herein we report the chemical synthesis and biological activity of conjugates of fluconazole with (i) cell-penetrating peptides (CPP), namely TP10-NH2 and TP10-7-NH2, or (ii) antimicrobial peptides (AMP), such as LFcinB(2-11)-NH2, LFcinB[Nle1,11]-NH2 and HLopt2-NH2. Both constituents of produced conjugates (FLC and peptides) display different modes of antifungal activity and affect different molecular targets within fungal cells. CPPs serving as carrier peptides and are considered the fundamental part of an extensively developed concept of a drug delivery system; TP-10 is one of the most promising peptides in this family. These peptides effectively penetrate cell membrane. In our recent paper. we showed that TP10-NH2 can be successfully applied to design conjugates with antimicrobial activity. Modified fragments of bovine lactoferrin (LFcin) and human lactoferrin (HLopt2) are members of the AMP family, peptides that constitute an important segment of the bodys natural immunity against microorganisms.
Click to Show/Hide
|
||||
| Description |
The inactivity of FLCpOH confirmed that the drug derivative, used during the synthesis of our target compounds, did not exert any antifungal activity. All compounds displayed poor activity against the C. glabrata strain tested. Another identified advantage of FLCpOH-TP10-NH2 and FLCpOH-TP10-7-NH2 is their higher fungicidal potential compared to fluconazole. The MFC values against C. albicans reference strains were the same or only twice higher compared to MIC90. The high fungicidal activity of both conjugates was also confirmed with the kill-time assay. However, at least a 4 x MFC concentration was necessary to obtain an evident killing effect after 24 h of treatment.
Click to Show/Hide
|
||||
| In Vitro Model | Acute monocytic leukemia | MONO-MAC-6 cell | CVCL_1426 | ||
| Experiment 7 Reporting the Activity Data of This PDC | [1] | ||||
| Indication | <i>Candida spp</i> infection | ||||
| Efficacy Data | Minimum fungicidal concentration | > 250 μM | |||
| Administration Time | 24 h | ||||
| Evaluation Method | Antifungal activity of the tested compounds against yeast cells was determined in 96-well plates, with minor modifications (CLSI, 2008). Suspensions of the microorganisms were prepared by taking one loop of pure culture into sterile water and adjusting optical density to 0.1 at 660 nm wavelength and further 50-fold dilution in RPMI 1640 medium, resulting in cell concentration of approximately 2 104 CFU/m. Then, 100 μL of cells were added to the wells of a 96-well microtiter plate that contained 16-500 μM tested compounds and 16-500 μM of FLC or 0.2-62 μM AmB. The plates were incubated for 24 h at 37 °C. Each test was performed in triplicate. The final concentration of DMSO was ensured to be around 1% in all experiments. The MIC90 was defined as the lowest concentration of drug showing no visible growth; MIC50 was defined as the concentration that yielded at least 50% growth inhibition when compared with the growth control well. For determination of minimum fungicidal concentrations (MFC), small aliquots of suspensions (around 10 μL) from each well were transferred using the pipette to YPD agar plates without inhibitors and incubated for 24 h at 37 °C. The MFC was defined as the lowest concentration of drug at which no growth of the colonies was observed. | ||||
| MOA of PDC |
Herein we report the chemical synthesis and biological activity of conjugates of fluconazole with (i) cell-penetrating peptides (CPP), namely TP10-NH2 and TP10-7-NH2, or (ii) antimicrobial peptides (AMP), such as LFcinB(2-11)-NH2, LFcinB[Nle1,11]-NH2 and HLopt2-NH2. Both constituents of produced conjugates (FLC and peptides) display different modes of antifungal activity and affect different molecular targets within fungal cells. CPPs serving as carrier peptides and are considered the fundamental part of an extensively developed concept of a drug delivery system; TP-10 is one of the most promising peptides in this family. These peptides effectively penetrate cell membrane. In our recent paper. we showed that TP10-NH2 can be successfully applied to design conjugates with antimicrobial activity. Modified fragments of bovine lactoferrin (LFcin) and human lactoferrin (HLopt2) are members of the AMP family, peptides that constitute an important segment of the bodys natural immunity against microorganisms.
Click to Show/Hide
|
||||
| Description |
The inactivity of FLCpOH confirmed that the drug derivative, used during the synthesis of our target compounds, did not exert any antifungal activity. All compounds displayed poor activity against the C. glabrata strain tested. Another identified advantage of FLCpOH-TP10-NH2 and FLCpOH-TP10-7-NH2 is their higher fungicidal potential compared to fluconazole. The MFC values against C. albicans reference strains were the same or only twice higher compared to MIC90. The high fungicidal activity of both conjugates was also confirmed with the kill-time assay. However, at least a 4 x MFC concentration was necessary to obtain an evident killing effect after 24 h of treatment.
Click to Show/Hide
|
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| In Vitro Model | Normal | MT4/HIV-1 cell | CVCL_RW54 | ||
| Experiment 8 Reporting the Activity Data of This PDC | [1] | ||||
| Indication | <i>Candida spp</i> infection | ||||
| Efficacy Data | Minimum fungicidal concentration | > 250 μM | |||
| Administration Time | 24 h | ||||
| Evaluation Method | Antifungal activity of the tested compounds against yeast cells was determined in 96-well plates, with minor modifications (CLSI, 2008). Suspensions of the microorganisms were prepared by taking one loop of pure culture into sterile water and adjusting optical density to 0.1 at 660 nm wavelength and further 50-fold dilution in RPMI 1640 medium, resulting in cell concentration of approximately 2 104 CFU/m. Then, 100 μL of cells were added to the wells of a 96-well microtiter plate that contained 16-500 μM tested compounds and 16-500 μM of FLC or 0.2-62 μM AmB. The plates were incubated for 24 h at 37 °C. Each test was performed in triplicate. The final concentration of DMSO was ensured to be around 1% in all experiments. The MIC90 was defined as the lowest concentration of drug showing no visible growth; MIC50 was defined as the concentration that yielded at least 50% growth inhibition when compared with the growth control well. For determination of minimum fungicidal concentrations (MFC), small aliquots of suspensions (around 10 μL) from each well were transferred using the pipette to YPD agar plates without inhibitors and incubated for 24 h at 37 °C. The MFC was defined as the lowest concentration of drug at which no growth of the colonies was observed. | ||||
| MOA of PDC |
Herein we report the chemical synthesis and biological activity of conjugates of fluconazole with (i) cell-penetrating peptides (CPP), namely TP10-NH2 and TP10-7-NH2, or (ii) antimicrobial peptides (AMP), such as LFcinB(2-11)-NH2, LFcinB[Nle1,11]-NH2 and HLopt2-NH2. Both constituents of produced conjugates (FLC and peptides) display different modes of antifungal activity and affect different molecular targets within fungal cells. CPPs serving as carrier peptides and are considered the fundamental part of an extensively developed concept of a drug delivery system; TP-10 is one of the most promising peptides in this family. These peptides effectively penetrate cell membrane. In our recent paper. we showed that TP10-NH2 can be successfully applied to design conjugates with antimicrobial activity. Modified fragments of bovine lactoferrin (LFcin) and human lactoferrin (HLopt2) are members of the AMP family, peptides that constitute an important segment of the bodys natural immunity against microorganisms.
Click to Show/Hide
|
||||
| Description |
The inactivity of FLCpOH confirmed that the drug derivative, used during the synthesis of our target compounds, did not exert any antifungal activity. All compounds displayed poor activity against the C. glabrata strain tested. Another identified advantage of FLCpOH-TP10-NH2 and FLCpOH-TP10-7-NH2 is their higher fungicidal potential compared to fluconazole. The MFC values against C. albicans reference strains were the same or only twice higher compared to MIC90. The high fungicidal activity of both conjugates was also confirmed with the kill-time assay. However, at least a 4 x MFC concentration was necessary to obtain an evident killing effect after 24 h of treatment.
Click to Show/Hide
|
||||
| In Vitro Model | Staphylococcus aureus infection | Staphylococcus aureus | 1280 | ||
| Experiment 9 Reporting the Activity Data of This PDC | [1] | ||||
| Indication | <i>Candida spp</i> infection | ||||
| Efficacy Data | Minimum fungicidal concentration |
250 μM
|
|||
| Administration Time | 24 h | ||||
| Evaluation Method | Antifungal activity of the tested compounds against yeast cells was determined in 96-well plates, with minor modifications (CLSI, 2008). Suspensions of the microorganisms were prepared by taking one loop of pure culture into sterile water and adjusting optical density to 0.1 at 660 nm wavelength and further 50-fold dilution in RPMI 1640 medium, resulting in cell concentration of approximately 2 104 CFU/m. Then, 100 μL of cells were added to the wells of a 96-well microtiter plate that contained 16-500 μM tested compounds and 16-500 μM of FLC or 0.2-62 μM AmB. The plates were incubated for 24 h at 37 °C. Each test was performed in triplicate. The final concentration of DMSO was ensured to be around 1% in all experiments. The MIC90 was defined as the lowest concentration of drug showing no visible growth; MIC50 was defined as the concentration that yielded at least 50% growth inhibition when compared with the growth control well. For determination of minimum fungicidal concentrations (MFC), small aliquots of suspensions (around 10 μL) from each well were transferred using the pipette to YPD agar plates without inhibitors and incubated for 24 h at 37 °C. The MFC was defined as the lowest concentration of drug at which no growth of the colonies was observed. | ||||
| MOA of PDC |
Herein we report the chemical synthesis and biological activity of conjugates of fluconazole with (i) cell-penetrating peptides (CPP), namely TP10-NH2 and TP10-7-NH2, or (ii) antimicrobial peptides (AMP), such as LFcinB(2-11)-NH2, LFcinB[Nle1,11]-NH2 and HLopt2-NH2. Both constituents of produced conjugates (FLC and peptides) display different modes of antifungal activity and affect different molecular targets within fungal cells. CPPs serving as carrier peptides and are considered the fundamental part of an extensively developed concept of a drug delivery system; TP-10 is one of the most promising peptides in this family. These peptides effectively penetrate cell membrane. In our recent paper. we showed that TP10-NH2 can be successfully applied to design conjugates with antimicrobial activity. Modified fragments of bovine lactoferrin (LFcin) and human lactoferrin (HLopt2) are members of the AMP family, peptides that constitute an important segment of the bodys natural immunity against microorganisms.
Click to Show/Hide
|
||||
| Description |
The inactivity of FLCpOH confirmed that the drug derivative, used during the synthesis of our target compounds, did not exert any antifungal activity. All compounds displayed poor activity against the C. glabrata strain tested. Another identified advantage of FLCpOH-TP10-NH2 and FLCpOH-TP10-7-NH2 is their higher fungicidal potential compared to fluconazole. The MFC values against C. albicans reference strains were the same or only twice higher compared to MIC90. The high fungicidal activity of both conjugates was also confirmed with the kill-time assay. However, at least a 4 x MFC concentration was necessary to obtain an evident killing effect after 24 h of treatment.
Click to Show/Hide
|
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| In Vitro Model | Breast adenocarcinoma | MDA-MB-231 (ACE+) cell | CVCL_0062 | ||
| Experiment 10 Reporting the Activity Data of This PDC | [1] | ||||
| Indication | <i>Candida spp</i> infection | ||||
| Efficacy Data | Minimum fungicidal concentration | > 250 μM | |||
| Administration Time | 24 h | ||||
| Evaluation Method | Antifungal activity of the tested compounds against yeast cells was determined in 96-well plates, with minor modifications (CLSI, 2008). Suspensions of the microorganisms were prepared by taking one loop of pure culture into sterile water and adjusting optical density to 0.1 at 660 nm wavelength and further 50-fold dilution in RPMI 1640 medium, resulting in cell concentration of approximately 2 104 CFU/m. Then, 100 μL of cells were added to the wells of a 96-well microtiter plate that contained 16-500 μM tested compounds and 16-500 μM of FLC or 0.2-62 μM AmB. The plates were incubated for 24 h at 37 °C. Each test was performed in triplicate. The final concentration of DMSO was ensured to be around 1% in all experiments. The MIC90 was defined as the lowest concentration of drug showing no visible growth; MIC50 was defined as the concentration that yielded at least 50% growth inhibition when compared with the growth control well. For determination of minimum fungicidal concentrations (MFC), small aliquots of suspensions (around 10 μL) from each well were transferred using the pipette to YPD agar plates without inhibitors and incubated for 24 h at 37 °C. The MFC was defined as the lowest concentration of drug at which no growth of the colonies was observed. | ||||
| MOA of PDC |
Herein we report the chemical synthesis and biological activity of conjugates of fluconazole with (i) cell-penetrating peptides (CPP), namely TP10-NH2 and TP10-7-NH2, or (ii) antimicrobial peptides (AMP), such as LFcinB(2-11)-NH2, LFcinB[Nle1,11]-NH2 and HLopt2-NH2. Both constituents of produced conjugates (FLC and peptides) display different modes of antifungal activity and affect different molecular targets within fungal cells. CPPs serving as carrier peptides and are considered the fundamental part of an extensively developed concept of a drug delivery system; TP-10 is one of the most promising peptides in this family. These peptides effectively penetrate cell membrane. In our recent paper. we showed that TP10-NH2 can be successfully applied to design conjugates with antimicrobial activity. Modified fragments of bovine lactoferrin (LFcin) and human lactoferrin (HLopt2) are members of the AMP family, peptides that constitute an important segment of the bodys natural immunity against microorganisms.
Click to Show/Hide
|
||||
| Description |
The inactivity of FLCpOH confirmed that the drug derivative, used during the synthesis of our target compounds, did not exert any antifungal activity. All compounds displayed poor activity against the C. glabrata strain tested. Another identified advantage of FLCpOH-TP10-NH2 and FLCpOH-TP10-7-NH2 is their higher fungicidal potential compared to fluconazole. The MFC values against C. albicans reference strains were the same or only twice higher compared to MIC90. The high fungicidal activity of both conjugates was also confirmed with the kill-time assay. However, at least a 4 x MFC concentration was necessary to obtain an evident killing effect after 24 h of treatment.
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| In Vitro Model | Breast cancer | MCF-7/6 cell | CVCL_W972 | ||
| Experiment 11 Reporting the Activity Data of This PDC | [1] | ||||
| Indication | <i>Candida spp</i> infection | ||||
| Efficacy Data | Minimum fungicidal concentration |
250 μM
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| Administration Time | 24 h | ||||
| Evaluation Method | Antifungal activity of the tested compounds against yeast cells was determined in 96-well plates, with minor modifications (CLSI, 2008). Suspensions of the microorganisms were prepared by taking one loop of pure culture into sterile water and adjusting optical density to 0.1 at 660 nm wavelength and further 50-fold dilution in RPMI 1640 medium, resulting in cell concentration of approximately 2 104 CFU/m. Then, 100 μL of cells were added to the wells of a 96-well microtiter plate that contained 16-500 μM tested compounds and 16-500 μM of FLC or 0.2-62 μM AmB. The plates were incubated for 24 h at 37 °C. Each test was performed in triplicate. The final concentration of DMSO was ensured to be around 1% in all experiments. The MIC90 was defined as the lowest concentration of drug showing no visible growth; MIC50 was defined as the concentration that yielded at least 50% growth inhibition when compared with the growth control well. For determination of minimum fungicidal concentrations (MFC), small aliquots of suspensions (around 10 μL) from each well were transferred using the pipette to YPD agar plates without inhibitors and incubated for 24 h at 37 °C. The MFC was defined as the lowest concentration of drug at which no growth of the colonies was observed. | ||||
| MOA of PDC |
Herein we report the chemical synthesis and biological activity of conjugates of fluconazole with (i) cell-penetrating peptides (CPP), namely TP10-NH2 and TP10-7-NH2, or (ii) antimicrobial peptides (AMP), such as LFcinB(2-11)-NH2, LFcinB[Nle1,11]-NH2 and HLopt2-NH2. Both constituents of produced conjugates (FLC and peptides) display different modes of antifungal activity and affect different molecular targets within fungal cells. CPPs serving as carrier peptides and are considered the fundamental part of an extensively developed concept of a drug delivery system; TP-10 is one of the most promising peptides in this family. These peptides effectively penetrate cell membrane. In our recent paper. we showed that TP10-NH2 can be successfully applied to design conjugates with antimicrobial activity. Modified fragments of bovine lactoferrin (LFcin) and human lactoferrin (HLopt2) are members of the AMP family, peptides that constitute an important segment of the bodys natural immunity against microorganisms.
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| Description |
The inactivity of FLCpOH confirmed that the drug derivative, used during the synthesis of our target compounds, did not exert any antifungal activity. All compounds displayed poor activity against the C. glabrata strain tested. Another identified advantage of FLCpOH-TP10-NH2 and FLCpOH-TP10-7-NH2 is their higher fungicidal potential compared to fluconazole. The MFC values against C. albicans reference strains were the same or only twice higher compared to MIC90. The high fungicidal activity of both conjugates was also confirmed with the kill-time assay. However, at least a 4 x MFC concentration was necessary to obtain an evident killing effect after 24 h of treatment.
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| In Vitro Model | Invasive breast carcinoma | MCF-7/ADR cell | CVCL_0031 | ||
| Experiment 12 Reporting the Activity Data of This PDC | [1] | ||||
| Indication | <i>Candida spp</i> infection | ||||
| Efficacy Data | Minimum fungicidal concentration | > 250 μM | |||
| Administration Time | 24 h | ||||
| Evaluation Method | Antifungal activity of the tested compounds against yeast cells was determined in 96-well plates, with minor modifications (CLSI, 2008). Suspensions of the microorganisms were prepared by taking one loop of pure culture into sterile water and adjusting optical density to 0.1 at 660 nm wavelength and further 50-fold dilution in RPMI 1640 medium, resulting in cell concentration of approximately 2 104 CFU/m. Then, 100 μL of cells were added to the wells of a 96-well microtiter plate that contained 16-500 μM tested compounds and 16-500 μM of FLC or 0.2-62 μM AmB. The plates were incubated for 24 h at 37 °C. Each test was performed in triplicate. The final concentration of DMSO was ensured to be around 1% in all experiments. The MIC90 was defined as the lowest concentration of drug showing no visible growth; MIC50 was defined as the concentration that yielded at least 50% growth inhibition when compared with the growth control well. For determination of minimum fungicidal concentrations (MFC), small aliquots of suspensions (around 10 μL) from each well were transferred using the pipette to YPD agar plates without inhibitors and incubated for 24 h at 37 °C. The MFC was defined as the lowest concentration of drug at which no growth of the colonies was observed. | ||||
| MOA of PDC |
Herein we report the chemical synthesis and biological activity of conjugates of fluconazole with (i) cell-penetrating peptides (CPP), namely TP10-NH2 and TP10-7-NH2, or (ii) antimicrobial peptides (AMP), such as LFcinB(2-11)-NH2, LFcinB[Nle1,11]-NH2 and HLopt2-NH2. Both constituents of produced conjugates (FLC and peptides) display different modes of antifungal activity and affect different molecular targets within fungal cells. CPPs serving as carrier peptides and are considered the fundamental part of an extensively developed concept of a drug delivery system; TP-10 is one of the most promising peptides in this family. These peptides effectively penetrate cell membrane. In our recent paper. we showed that TP10-NH2 can be successfully applied to design conjugates with antimicrobial activity. Modified fragments of bovine lactoferrin (LFcin) and human lactoferrin (HLopt2) are members of the AMP family, peptides that constitute an important segment of the bodys natural immunity against microorganisms.
Click to Show/Hide
|
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| Description |
The inactivity of FLCpOH confirmed that the drug derivative, used during the synthesis of our target compounds, did not exert any antifungal activity. All compounds displayed poor activity against the C. glabrata strain tested. Another identified advantage of FLCpOH-TP10-NH2 and FLCpOH-TP10-7-NH2 is their higher fungicidal potential compared to fluconazole. The MFC values against C. albicans reference strains were the same or only twice higher compared to MIC90. The high fungicidal activity of both conjugates was also confirmed with the kill-time assay. However, at least a 4 x MFC concentration was necessary to obtain an evident killing effect after 24 h of treatment.
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| In Vitro Model | Invasive ductal carcinoma | MCF7/PTX cell | CVCL_C5RS | ||
| Experiment 13 Reporting the Activity Data of This PDC | [1] | ||||
| Indication | <i>Candida spp</i> infection | ||||
| Efficacy Data | Minimum fungicidal concentration |
250 μM
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| Administration Time | 24 h | ||||
| Evaluation Method | Antifungal activity of the tested compounds against yeast cells was determined in 96-well plates, with minor modifications (CLSI, 2008). Suspensions of the microorganisms were prepared by taking one loop of pure culture into sterile water and adjusting optical density to 0.1 at 660 nm wavelength and further 50-fold dilution in RPMI 1640 medium, resulting in cell concentration of approximately 2 104 CFU/m. Then, 100 μL of cells were added to the wells of a 96-well microtiter plate that contained 16-500 μM tested compounds and 16-500 μM of FLC or 0.2-62 μM AmB. The plates were incubated for 24 h at 37 °C. Each test was performed in triplicate. The final concentration of DMSO was ensured to be around 1% in all experiments. The MIC90 was defined as the lowest concentration of drug showing no visible growth; MIC50 was defined as the concentration that yielded at least 50% growth inhibition when compared with the growth control well. For determination of minimum fungicidal concentrations (MFC), small aliquots of suspensions (around 10 μL) from each well were transferred using the pipette to YPD agar plates without inhibitors and incubated for 24 h at 37 °C. The MFC was defined as the lowest concentration of drug at which no growth of the colonies was observed. | ||||
| MOA of PDC |
Herein we report the chemical synthesis and biological activity of conjugates of fluconazole with (i) cell-penetrating peptides (CPP), namely TP10-NH2 and TP10-7-NH2, or (ii) antimicrobial peptides (AMP), such as LFcinB(2-11)-NH2, LFcinB[Nle1,11]-NH2 and HLopt2-NH2. Both constituents of produced conjugates (FLC and peptides) display different modes of antifungal activity and affect different molecular targets within fungal cells. CPPs serving as carrier peptides and are considered the fundamental part of an extensively developed concept of a drug delivery system; TP-10 is one of the most promising peptides in this family. These peptides effectively penetrate cell membrane. In our recent paper. we showed that TP10-NH2 can be successfully applied to design conjugates with antimicrobial activity. Modified fragments of bovine lactoferrin (LFcin) and human lactoferrin (HLopt2) are members of the AMP family, peptides that constitute an important segment of the bodys natural immunity against microorganisms.
Click to Show/Hide
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| Description |
The inactivity of FLCpOH confirmed that the drug derivative, used during the synthesis of our target compounds, did not exert any antifungal activity. All compounds displayed poor activity against the C. glabrata strain tested. Another identified advantage of FLCpOH-TP10-NH2 and FLCpOH-TP10-7-NH2 is their higher fungicidal potential compared to fluconazole. The MFC values against C. albicans reference strains were the same or only twice higher compared to MIC90. The high fungicidal activity of both conjugates was also confirmed with the kill-time assay. However, at least a 4 x MFC concentration was necessary to obtain an evident killing effect after 24 h of treatment.
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| In Vitro Model | Breast adenocarcinoma | MDA-MB-231 (KRAS G13D) cell | CVCL_0062 | ||
| Experiment 14 Reporting the Activity Data of This PDC | [1] | ||||
| Indication | <i>Candida spp</i> infection | ||||
| Efficacy Data | Minimum fungicidal concentration | > 250 μM | |||
| Administration Time | 24 h | ||||
| Evaluation Method | Antifungal activity of the tested compounds against yeast cells was determined in 96-well plates, with minor modifications (CLSI, 2008). Suspensions of the microorganisms were prepared by taking one loop of pure culture into sterile water and adjusting optical density to 0.1 at 660 nm wavelength and further 50-fold dilution in RPMI 1640 medium, resulting in cell concentration of approximately 2 104 CFU/m. Then, 100 μL of cells were added to the wells of a 96-well microtiter plate that contained 16-500 μM tested compounds and 16-500 μM of FLC or 0.2-62 μM AmB. The plates were incubated for 24 h at 37 °C. Each test was performed in triplicate. The final concentration of DMSO was ensured to be around 1% in all experiments. The MIC90 was defined as the lowest concentration of drug showing no visible growth; MIC50 was defined as the concentration that yielded at least 50% growth inhibition when compared with the growth control well. For determination of minimum fungicidal concentrations (MFC), small aliquots of suspensions (around 10 μL) from each well were transferred using the pipette to YPD agar plates without inhibitors and incubated for 24 h at 37 °C. The MFC was defined as the lowest concentration of drug at which no growth of the colonies was observed. | ||||
| MOA of PDC |
Herein we report the chemical synthesis and biological activity of conjugates of fluconazole with (i) cell-penetrating peptides (CPP), namely TP10-NH2 and TP10-7-NH2, or (ii) antimicrobial peptides (AMP), such as LFcinB(2-11)-NH2, LFcinB[Nle1,11]-NH2 and HLopt2-NH2. Both constituents of produced conjugates (FLC and peptides) display different modes of antifungal activity and affect different molecular targets within fungal cells. CPPs serving as carrier peptides and are considered the fundamental part of an extensively developed concept of a drug delivery system; TP-10 is one of the most promising peptides in this family. These peptides effectively penetrate cell membrane. In our recent paper. we showed that TP10-NH2 can be successfully applied to design conjugates with antimicrobial activity. Modified fragments of bovine lactoferrin (LFcin) and human lactoferrin (HLopt2) are members of the AMP family, peptides that constitute an important segment of the bodys natural immunity against microorganisms.
Click to Show/Hide
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| Description |
The inactivity of FLCpOH confirmed that the drug derivative, used during the synthesis of our target compounds, did not exert any antifungal activity. All compounds displayed poor activity against the C. glabrata strain tested. Another identified advantage of FLCpOH-TP10-NH2 and FLCpOH-TP10-7-NH2 is their higher fungicidal potential compared to fluconazole. The MFC values against C. albicans reference strains were the same or only twice higher compared to MIC90. The high fungicidal activity of both conjugates was also confirmed with the kill-time assay. However, at least a 4 x MFC concentration was necessary to obtain an evident killing effect after 24 h of treatment.
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| In Vitro Model | Uterine sarcoma | MES-SA/Dx5 cell | CVCL_2598 | ||
| Experiment 15 Reporting the Activity Data of This PDC | [1] | ||||
| Indication | <i>Candida spp</i> infection | ||||
| Efficacy Data | Half minimum inhibitory concentration (MIC90) |
125 μM
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| Administration Time | 24 h | ||||
| Evaluation Method | Antifungal activity of the tested compounds against yeast cells was determined in 96-well plates, with minor modifications (CLSI, 2008). Suspensions of the microorganisms were prepared by taking one loop of pure culture into sterile water and adjusting optical density to 0.1 at 660 nm wavelength and further 50-fold dilution in RPMI 1640 medium, resulting in cell concentration of approximately 2 104 CFU/m. Then, 100 μL of cells were added to the wells of a 96-well microtiter plate that contained 16-500 μM tested compounds and 16-500 μM of FLC or 0.2-62 μM AmB. The plates were incubated for 24 h at 37 °C. Each test was performed in triplicate. The final concentration of DMSO was ensured to be around 1% in all experiments. The MIC90 was defined as the lowest concentration of drug showing no visible growth; MIC50 was defined as the concentration that yielded at least 50% growth inhibition when compared with the growth control well. For determination of minimum fungicidal concentrations (MFC), small aliquots of suspensions (around 10 μL) from each well were transferred using the pipette to YPD agar plates without inhibitors and incubated for 24 h at 37 °C. The MFC was defined as the lowest concentration of drug at which no growth of the colonies was observed. | ||||
| MOA of PDC |
Herein we report the chemical synthesis and biological activity of conjugates of fluconazole with (i) cell-penetrating peptides (CPP), namely TP10-NH2 and TP10-7-NH2, or (ii) antimicrobial peptides (AMP), such as LFcinB(2-11)-NH2, LFcinB[Nle1,11]-NH2 and HLopt2-NH2. Both constituents of produced conjugates (FLC and peptides) display different modes of antifungal activity and affect different molecular targets within fungal cells. CPPs serving as carrier peptides and are considered the fundamental part of an extensively developed concept of a drug delivery system; TP-10 is one of the most promising peptides in this family. These peptides effectively penetrate cell membrane. In our recent paper. we showed that TP10-NH2 can be successfully applied to design conjugates with antimicrobial activity. Modified fragments of bovine lactoferrin (LFcin) and human lactoferrin (HLopt2) are members of the AMP family, peptides that constitute an important segment of the bodys natural immunity against microorganisms.
Click to Show/Hide
|
||||
| Description |
The inactivity of FLCpOH confirmed that the drug derivative, used during the synthesis of our target compounds, did not exert any antifungal activity. All compounds displayed poor activity against the C. glabrata strain tested. Another identified advantage of FLCpOH-TP10-NH2 and FLCpOH-TP10-7-NH2 is their higher fungicidal potential compared to fluconazole. The MFC values against C. albicans reference strains were the same or only twice higher compared to MIC90. The high fungicidal activity of both conjugates was also confirmed with the kill-time assay. However, at least a 4 x MFC concentration was necessary to obtain an evident killing effect after 24 h of treatment.
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| In Vitro Model | Staphylococcus aureus infection | Staphylococcus aureus | 1280 | ||
| Experiment 16 Reporting the Activity Data of This PDC | [1] | ||||
| Indication | <i>Candida spp</i> infection | ||||
| Efficacy Data | Half minimum inhibitory concentration (MIC90) | > 250 μM | |||
| Administration Time | 24 h | ||||
| Evaluation Method | Antifungal activity of the tested compounds against yeast cells was determined in 96-well plates, with minor modifications (CLSI, 2008). Suspensions of the microorganisms were prepared by taking one loop of pure culture into sterile water and adjusting optical density to 0.1 at 660 nm wavelength and further 50-fold dilution in RPMI 1640 medium, resulting in cell concentration of approximately 2 104 CFU/m. Then, 100 μL of cells were added to the wells of a 96-well microtiter plate that contained 16-500 μM tested compounds and 16-500 μM of FLC or 0.2-62 μM AmB. The plates were incubated for 24 h at 37 °C. Each test was performed in triplicate. The final concentration of DMSO was ensured to be around 1% in all experiments. The MIC90 was defined as the lowest concentration of drug showing no visible growth; MIC50 was defined as the concentration that yielded at least 50% growth inhibition when compared with the growth control well. For determination of minimum fungicidal concentrations (MFC), small aliquots of suspensions (around 10 μL) from each well were transferred using the pipette to YPD agar plates without inhibitors and incubated for 24 h at 37 °C. The MFC was defined as the lowest concentration of drug at which no growth of the colonies was observed. | ||||
| MOA of PDC |
Herein we report the chemical synthesis and biological activity of conjugates of fluconazole with (i) cell-penetrating peptides (CPP), namely TP10-NH2 and TP10-7-NH2, or (ii) antimicrobial peptides (AMP), such as LFcinB(2-11)-NH2, LFcinB[Nle1,11]-NH2 and HLopt2-NH2. Both constituents of produced conjugates (FLC and peptides) display different modes of antifungal activity and affect different molecular targets within fungal cells. CPPs serving as carrier peptides and are considered the fundamental part of an extensively developed concept of a drug delivery system; TP-10 is one of the most promising peptides in this family. These peptides effectively penetrate cell membrane. In our recent paper. we showed that TP10-NH2 can be successfully applied to design conjugates with antimicrobial activity. Modified fragments of bovine lactoferrin (LFcin) and human lactoferrin (HLopt2) are members of the AMP family, peptides that constitute an important segment of the bodys natural immunity against microorganisms.
Click to Show/Hide
|
||||
| Description |
The inactivity of FLCpOH confirmed that the drug derivative, used during the synthesis of our target compounds, did not exert any antifungal activity. All compounds displayed poor activity against the C. glabrata strain tested. Another identified advantage of FLCpOH-TP10-NH2 and FLCpOH-TP10-7-NH2 is their higher fungicidal potential compared to fluconazole. The MFC values against C. albicans reference strains were the same or only twice higher compared to MIC90. The high fungicidal activity of both conjugates was also confirmed with the kill-time assay. However, at least a 4 x MFC concentration was necessary to obtain an evident killing effect after 24 h of treatment.
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| In Vitro Model | Mouse colon adenocarcinoma | MC-38 cell | CVCL_B288 | ||
| Experiment 17 Reporting the Activity Data of This PDC | [1] | ||||
| Indication | <i>Candida spp</i> infection | ||||
| Efficacy Data | Half minimum inhibitory concentration (MIC90) | > 250 μM | |||
| Administration Time | 24 h | ||||
| Evaluation Method | Antifungal activity of the tested compounds against yeast cells was determined in 96-well plates, with minor modifications (CLSI, 2008). Suspensions of the microorganisms were prepared by taking one loop of pure culture into sterile water and adjusting optical density to 0.1 at 660 nm wavelength and further 50-fold dilution in RPMI 1640 medium, resulting in cell concentration of approximately 2 104 CFU/m. Then, 100 μL of cells were added to the wells of a 96-well microtiter plate that contained 16-500 μM tested compounds and 16-500 μM of FLC or 0.2-62 μM AmB. The plates were incubated for 24 h at 37 °C. Each test was performed in triplicate. The final concentration of DMSO was ensured to be around 1% in all experiments. The MIC90 was defined as the lowest concentration of drug showing no visible growth; MIC50 was defined as the concentration that yielded at least 50% growth inhibition when compared with the growth control well. For determination of minimum fungicidal concentrations (MFC), small aliquots of suspensions (around 10 μL) from each well were transferred using the pipette to YPD agar plates without inhibitors and incubated for 24 h at 37 °C. The MFC was defined as the lowest concentration of drug at which no growth of the colonies was observed. | ||||
| MOA of PDC |
Herein we report the chemical synthesis and biological activity of conjugates of fluconazole with (i) cell-penetrating peptides (CPP), namely TP10-NH2 and TP10-7-NH2, or (ii) antimicrobial peptides (AMP), such as LFcinB(2-11)-NH2, LFcinB[Nle1,11]-NH2 and HLopt2-NH2. Both constituents of produced conjugates (FLC and peptides) display different modes of antifungal activity and affect different molecular targets within fungal cells. CPPs serving as carrier peptides and are considered the fundamental part of an extensively developed concept of a drug delivery system; TP-10 is one of the most promising peptides in this family. These peptides effectively penetrate cell membrane. In our recent paper. we showed that TP10-NH2 can be successfully applied to design conjugates with antimicrobial activity. Modified fragments of bovine lactoferrin (LFcin) and human lactoferrin (HLopt2) are members of the AMP family, peptides that constitute an important segment of the bodys natural immunity against microorganisms.
Click to Show/Hide
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| Description |
The inactivity of FLCpOH confirmed that the drug derivative, used during the synthesis of our target compounds, did not exert any antifungal activity. All compounds displayed poor activity against the C. glabrata strain tested. Another identified advantage of FLCpOH-TP10-NH2 and FLCpOH-TP10-7-NH2 is their higher fungicidal potential compared to fluconazole. The MFC values against C. albicans reference strains were the same or only twice higher compared to MIC90. The high fungicidal activity of both conjugates was also confirmed with the kill-time assay. However, at least a 4 x MFC concentration was necessary to obtain an evident killing effect after 24 h of treatment.
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| In Vitro Model | Klebsiella pneumoniae infection | Klebsiella pneumoniae | 573 | ||
| Experiment 18 Reporting the Activity Data of This PDC | [1] | ||||
| Indication | <i>Candida spp</i> infection | ||||
| Efficacy Data | Half minimum inhibitory concentration (MIC90) |
250 μM
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| Administration Time | 24 h | ||||
| Evaluation Method | Antifungal activity of the tested compounds against yeast cells was determined in 96-well plates, with minor modifications (CLSI, 2008). Suspensions of the microorganisms were prepared by taking one loop of pure culture into sterile water and adjusting optical density to 0.1 at 660 nm wavelength and further 50-fold dilution in RPMI 1640 medium, resulting in cell concentration of approximately 2 104 CFU/m. Then, 100 μL of cells were added to the wells of a 96-well microtiter plate that contained 16-500 μM tested compounds and 16-500 μM of FLC or 0.2-62 μM AmB. The plates were incubated for 24 h at 37 °C. Each test was performed in triplicate. The final concentration of DMSO was ensured to be around 1% in all experiments. The MIC90 was defined as the lowest concentration of drug showing no visible growth; MIC50 was defined as the concentration that yielded at least 50% growth inhibition when compared with the growth control well. For determination of minimum fungicidal concentrations (MFC), small aliquots of suspensions (around 10 μL) from each well were transferred using the pipette to YPD agar plates without inhibitors and incubated for 24 h at 37 °C. The MFC was defined as the lowest concentration of drug at which no growth of the colonies was observed. | ||||
| MOA of PDC |
Herein we report the chemical synthesis and biological activity of conjugates of fluconazole with (i) cell-penetrating peptides (CPP), namely TP10-NH2 and TP10-7-NH2, or (ii) antimicrobial peptides (AMP), such as LFcinB(2-11)-NH2, LFcinB[Nle1,11]-NH2 and HLopt2-NH2. Both constituents of produced conjugates (FLC and peptides) display different modes of antifungal activity and affect different molecular targets within fungal cells. CPPs serving as carrier peptides and are considered the fundamental part of an extensively developed concept of a drug delivery system; TP-10 is one of the most promising peptides in this family. These peptides effectively penetrate cell membrane. In our recent paper. we showed that TP10-NH2 can be successfully applied to design conjugates with antimicrobial activity. Modified fragments of bovine lactoferrin (LFcin) and human lactoferrin (HLopt2) are members of the AMP family, peptides that constitute an important segment of the bodys natural immunity against microorganisms.
Click to Show/Hide
|
||||
| Description |
The inactivity of FLCpOH confirmed that the drug derivative, used during the synthesis of our target compounds, did not exert any antifungal activity. All compounds displayed poor activity against the C. glabrata strain tested. Another identified advantage of FLCpOH-TP10-NH2 and FLCpOH-TP10-7-NH2 is their higher fungicidal potential compared to fluconazole. The MFC values against C. albicans reference strains were the same or only twice higher compared to MIC90. The high fungicidal activity of both conjugates was also confirmed with the kill-time assay. However, at least a 4 x MFC concentration was necessary to obtain an evident killing effect after 24 h of treatment.
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| In Vitro Model | Hepatocellular carcinoma | MHCC97H cell | CVCL_4972 | ||
| Experiment 19 Reporting the Activity Data of This PDC | [1] | ||||
| Indication | <i>Candida spp</i> infection | ||||
| Efficacy Data | Half minimum inhibitory concentration (MIC90) | > 250 μM | |||
| Administration Time | 24 h | ||||
| Evaluation Method | Antifungal activity of the tested compounds against yeast cells was determined in 96-well plates, with minor modifications (CLSI, 2008). Suspensions of the microorganisms were prepared by taking one loop of pure culture into sterile water and adjusting optical density to 0.1 at 660 nm wavelength and further 50-fold dilution in RPMI 1640 medium, resulting in cell concentration of approximately 2 104 CFU/m. Then, 100 μL of cells were added to the wells of a 96-well microtiter plate that contained 16-500 μM tested compounds and 16-500 μM of FLC or 0.2-62 μM AmB. The plates were incubated for 24 h at 37 °C. Each test was performed in triplicate. The final concentration of DMSO was ensured to be around 1% in all experiments. The MIC90 was defined as the lowest concentration of drug showing no visible growth; MIC50 was defined as the concentration that yielded at least 50% growth inhibition when compared with the growth control well. For determination of minimum fungicidal concentrations (MFC), small aliquots of suspensions (around 10 μL) from each well were transferred using the pipette to YPD agar plates without inhibitors and incubated for 24 h at 37 °C. The MFC was defined as the lowest concentration of drug at which no growth of the colonies was observed. | ||||
| MOA of PDC |
Herein we report the chemical synthesis and biological activity of conjugates of fluconazole with (i) cell-penetrating peptides (CPP), namely TP10-NH2 and TP10-7-NH2, or (ii) antimicrobial peptides (AMP), such as LFcinB(2-11)-NH2, LFcinB[Nle1,11]-NH2 and HLopt2-NH2. Both constituents of produced conjugates (FLC and peptides) display different modes of antifungal activity and affect different molecular targets within fungal cells. CPPs serving as carrier peptides and are considered the fundamental part of an extensively developed concept of a drug delivery system; TP-10 is one of the most promising peptides in this family. These peptides effectively penetrate cell membrane. In our recent paper. we showed that TP10-NH2 can be successfully applied to design conjugates with antimicrobial activity. Modified fragments of bovine lactoferrin (LFcin) and human lactoferrin (HLopt2) are members of the AMP family, peptides that constitute an important segment of the bodys natural immunity against microorganisms.
Click to Show/Hide
|
||||
| Description |
The inactivity of FLCpOH confirmed that the drug derivative, used during the synthesis of our target compounds, did not exert any antifungal activity. All compounds displayed poor activity against the C. glabrata strain tested. Another identified advantage of FLCpOH-TP10-NH2 and FLCpOH-TP10-7-NH2 is their higher fungicidal potential compared to fluconazole. The MFC values against C. albicans reference strains were the same or only twice higher compared to MIC90. The high fungicidal activity of both conjugates was also confirmed with the kill-time assay. However, at least a 4 x MFC concentration was necessary to obtain an evident killing effect after 24 h of treatment.
Click to Show/Hide
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| In Vitro Model | Pancreatic ductal adenocarcinoma | MIA PaCa-2 (KRAS G12C) cell | CVCL_0428 | ||
| Experiment 20 Reporting the Activity Data of This PDC | [1] | ||||
| Indication | <i>Candida spp</i> infection | ||||
| Efficacy Data | Half minimum inhibitory concentration (MIC90) | > 250 μM | |||
| Administration Time | 24 h | ||||
| Evaluation Method | Antifungal activity of the tested compounds against yeast cells was determined in 96-well plates, with minor modifications (CLSI, 2008). Suspensions of the microorganisms were prepared by taking one loop of pure culture into sterile water and adjusting optical density to 0.1 at 660 nm wavelength and further 50-fold dilution in RPMI 1640 medium, resulting in cell concentration of approximately 2 104 CFU/m. Then, 100 μL of cells were added to the wells of a 96-well microtiter plate that contained 16-500 μM tested compounds and 16-500 μM of FLC or 0.2-62 μM AmB. The plates were incubated for 24 h at 37 °C. Each test was performed in triplicate. The final concentration of DMSO was ensured to be around 1% in all experiments. The MIC90 was defined as the lowest concentration of drug showing no visible growth; MIC50 was defined as the concentration that yielded at least 50% growth inhibition when compared with the growth control well. For determination of minimum fungicidal concentrations (MFC), small aliquots of suspensions (around 10 μL) from each well were transferred using the pipette to YPD agar plates without inhibitors and incubated for 24 h at 37 °C. The MFC was defined as the lowest concentration of drug at which no growth of the colonies was observed. | ||||
| MOA of PDC |
Herein we report the chemical synthesis and biological activity of conjugates of fluconazole with (i) cell-penetrating peptides (CPP), namely TP10-NH2 and TP10-7-NH2, or (ii) antimicrobial peptides (AMP), such as LFcinB(2-11)-NH2, LFcinB[Nle1,11]-NH2 and HLopt2-NH2. Both constituents of produced conjugates (FLC and peptides) display different modes of antifungal activity and affect different molecular targets within fungal cells. CPPs serving as carrier peptides and are considered the fundamental part of an extensively developed concept of a drug delivery system; TP-10 is one of the most promising peptides in this family. These peptides effectively penetrate cell membrane. In our recent paper. we showed that TP10-NH2 can be successfully applied to design conjugates with antimicrobial activity. Modified fragments of bovine lactoferrin (LFcin) and human lactoferrin (HLopt2) are members of the AMP family, peptides that constitute an important segment of the bodys natural immunity against microorganisms.
Click to Show/Hide
|
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| Description |
The inactivity of FLCpOH confirmed that the drug derivative, used during the synthesis of our target compounds, did not exert any antifungal activity. All compounds displayed poor activity against the C. glabrata strain tested. Another identified advantage of FLCpOH-TP10-NH2 and FLCpOH-TP10-7-NH2 is their higher fungicidal potential compared to fluconazole. The MFC values against C. albicans reference strains were the same or only twice higher compared to MIC90. The high fungicidal activity of both conjugates was also confirmed with the kill-time assay. However, at least a 4 x MFC concentration was necessary to obtain an evident killing effect after 24 h of treatment.
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| In Vitro Model | Acute monocytic leukemia | MONO-MAC-6 cell | CVCL_1426 | ||
| Experiment 21 Reporting the Activity Data of This PDC | [1] | ||||
| Indication | <i>Candida spp</i> infection | ||||
| Efficacy Data | Half minimum inhibitory concentration (MIC90) | > 250 μM | |||
| Administration Time | 24 h | ||||
| Evaluation Method | Antifungal activity of the tested compounds against yeast cells was determined in 96-well plates, with minor modifications (CLSI, 2008). Suspensions of the microorganisms were prepared by taking one loop of pure culture into sterile water and adjusting optical density to 0.1 at 660 nm wavelength and further 50-fold dilution in RPMI 1640 medium, resulting in cell concentration of approximately 2 104 CFU/m. Then, 100 μL of cells were added to the wells of a 96-well microtiter plate that contained 16-500 μM tested compounds and 16-500 μM of FLC or 0.2-62 μM AmB. The plates were incubated for 24 h at 37 °C. Each test was performed in triplicate. The final concentration of DMSO was ensured to be around 1% in all experiments. The MIC90 was defined as the lowest concentration of drug showing no visible growth; MIC50 was defined as the concentration that yielded at least 50% growth inhibition when compared with the growth control well. For determination of minimum fungicidal concentrations (MFC), small aliquots of suspensions (around 10 μL) from each well were transferred using the pipette to YPD agar plates without inhibitors and incubated for 24 h at 37 °C. The MFC was defined as the lowest concentration of drug at which no growth of the colonies was observed. | ||||
| MOA of PDC |
Herein we report the chemical synthesis and biological activity of conjugates of fluconazole with (i) cell-penetrating peptides (CPP), namely TP10-NH2 and TP10-7-NH2, or (ii) antimicrobial peptides (AMP), such as LFcinB(2-11)-NH2, LFcinB[Nle1,11]-NH2 and HLopt2-NH2. Both constituents of produced conjugates (FLC and peptides) display different modes of antifungal activity and affect different molecular targets within fungal cells. CPPs serving as carrier peptides and are considered the fundamental part of an extensively developed concept of a drug delivery system; TP-10 is one of the most promising peptides in this family. These peptides effectively penetrate cell membrane. In our recent paper. we showed that TP10-NH2 can be successfully applied to design conjugates with antimicrobial activity. Modified fragments of bovine lactoferrin (LFcin) and human lactoferrin (HLopt2) are members of the AMP family, peptides that constitute an important segment of the bodys natural immunity against microorganisms.
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| Description |
The inactivity of FLCpOH confirmed that the drug derivative, used during the synthesis of our target compounds, did not exert any antifungal activity. All compounds displayed poor activity against the C. glabrata strain tested. Another identified advantage of FLCpOH-TP10-NH2 and FLCpOH-TP10-7-NH2 is their higher fungicidal potential compared to fluconazole. The MFC values against C. albicans reference strains were the same or only twice higher compared to MIC90. The high fungicidal activity of both conjugates was also confirmed with the kill-time assay. However, at least a 4 x MFC concentration was necessary to obtain an evident killing effect after 24 h of treatment.
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| In Vitro Model | Normal | MT4/HIV-1 cell | CVCL_RW54 | ||
| Experiment 22 Reporting the Activity Data of This PDC | [1] | ||||
| Indication | <i>Candida spp</i> infection | ||||
| Efficacy Data | Half minimum inhibitory concentration (MIC90) | > 250 μM | |||
| Administration Time | 24 h | ||||
| Evaluation Method | Antifungal activity of the tested compounds against yeast cells was determined in 96-well plates, with minor modifications (CLSI, 2008). Suspensions of the microorganisms were prepared by taking one loop of pure culture into sterile water and adjusting optical density to 0.1 at 660 nm wavelength and further 50-fold dilution in RPMI 1640 medium, resulting in cell concentration of approximately 2 104 CFU/m. Then, 100 μL of cells were added to the wells of a 96-well microtiter plate that contained 16-500 μM tested compounds and 16-500 μM of FLC or 0.2-62 μM AmB. The plates were incubated for 24 h at 37 °C. Each test was performed in triplicate. The final concentration of DMSO was ensured to be around 1% in all experiments. The MIC90 was defined as the lowest concentration of drug showing no visible growth; MIC50 was defined as the concentration that yielded at least 50% growth inhibition when compared with the growth control well. For determination of minimum fungicidal concentrations (MFC), small aliquots of suspensions (around 10 μL) from each well were transferred using the pipette to YPD agar plates without inhibitors and incubated for 24 h at 37 °C. The MFC was defined as the lowest concentration of drug at which no growth of the colonies was observed. | ||||
| MOA of PDC |
Herein we report the chemical synthesis and biological activity of conjugates of fluconazole with (i) cell-penetrating peptides (CPP), namely TP10-NH2 and TP10-7-NH2, or (ii) antimicrobial peptides (AMP), such as LFcinB(2-11)-NH2, LFcinB[Nle1,11]-NH2 and HLopt2-NH2. Both constituents of produced conjugates (FLC and peptides) display different modes of antifungal activity and affect different molecular targets within fungal cells. CPPs serving as carrier peptides and are considered the fundamental part of an extensively developed concept of a drug delivery system; TP-10 is one of the most promising peptides in this family. These peptides effectively penetrate cell membrane. In our recent paper. we showed that TP10-NH2 can be successfully applied to design conjugates with antimicrobial activity. Modified fragments of bovine lactoferrin (LFcin) and human lactoferrin (HLopt2) are members of the AMP family, peptides that constitute an important segment of the bodys natural immunity against microorganisms.
Click to Show/Hide
|
||||
| Description |
The inactivity of FLCpOH confirmed that the drug derivative, used during the synthesis of our target compounds, did not exert any antifungal activity. All compounds displayed poor activity against the C. glabrata strain tested. Another identified advantage of FLCpOH-TP10-NH2 and FLCpOH-TP10-7-NH2 is their higher fungicidal potential compared to fluconazole. The MFC values against C. albicans reference strains were the same or only twice higher compared to MIC90. The high fungicidal activity of both conjugates was also confirmed with the kill-time assay. However, at least a 4 x MFC concentration was necessary to obtain an evident killing effect after 24 h of treatment.
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| In Vitro Model | Staphylococcus aureus infection | Staphylococcus aureus | 1280 | ||
| Experiment 23 Reporting the Activity Data of This PDC | [1] | ||||
| Indication | <i>Candida spp</i> infection | ||||
| Efficacy Data | Half minimum inhibitory concentration (MIC90) | > 250 μM | |||
| Administration Time | 24 h | ||||
| Evaluation Method | Antifungal activity of the tested compounds against yeast cells was determined in 96-well plates, with minor modifications (CLSI, 2008). Suspensions of the microorganisms were prepared by taking one loop of pure culture into sterile water and adjusting optical density to 0.1 at 660 nm wavelength and further 50-fold dilution in RPMI 1640 medium, resulting in cell concentration of approximately 2 104 CFU/m. Then, 100 μL of cells were added to the wells of a 96-well microtiter plate that contained 16-500 μM tested compounds and 16-500 μM of FLC or 0.2-62 μM AmB. The plates were incubated for 24 h at 37 °C. Each test was performed in triplicate. The final concentration of DMSO was ensured to be around 1% in all experiments. The MIC90 was defined as the lowest concentration of drug showing no visible growth; MIC50 was defined as the concentration that yielded at least 50% growth inhibition when compared with the growth control well. For determination of minimum fungicidal concentrations (MFC), small aliquots of suspensions (around 10 μL) from each well were transferred using the pipette to YPD agar plates without inhibitors and incubated for 24 h at 37 °C. The MFC was defined as the lowest concentration of drug at which no growth of the colonies was observed. | ||||
| MOA of PDC |
Herein we report the chemical synthesis and biological activity of conjugates of fluconazole with (i) cell-penetrating peptides (CPP), namely TP10-NH2 and TP10-7-NH2, or (ii) antimicrobial peptides (AMP), such as LFcinB(2-11)-NH2, LFcinB[Nle1,11]-NH2 and HLopt2-NH2. Both constituents of produced conjugates (FLC and peptides) display different modes of antifungal activity and affect different molecular targets within fungal cells. CPPs serving as carrier peptides and are considered the fundamental part of an extensively developed concept of a drug delivery system; TP-10 is one of the most promising peptides in this family. These peptides effectively penetrate cell membrane. In our recent paper. we showed that TP10-NH2 can be successfully applied to design conjugates with antimicrobial activity. Modified fragments of bovine lactoferrin (LFcin) and human lactoferrin (HLopt2) are members of the AMP family, peptides that constitute an important segment of the bodys natural immunity against microorganisms.
Click to Show/Hide
|
||||
| Description |
The inactivity of FLCpOH confirmed that the drug derivative, used during the synthesis of our target compounds, did not exert any antifungal activity. All compounds displayed poor activity against the C. glabrata strain tested. Another identified advantage of FLCpOH-TP10-NH2 and FLCpOH-TP10-7-NH2 is their higher fungicidal potential compared to fluconazole. The MFC values against C. albicans reference strains were the same or only twice higher compared to MIC90. The high fungicidal activity of both conjugates was also confirmed with the kill-time assay. However, at least a 4 x MFC concentration was necessary to obtain an evident killing effect after 24 h of treatment.
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| In Vitro Model | Breast adenocarcinoma | MDA-MB-231 (ACE+) cell | CVCL_0062 | ||
| Experiment 24 Reporting the Activity Data of This PDC | [1] | ||||
| Indication | <i>Candida spp</i> infection | ||||
| Efficacy Data | Half minimum inhibitory concentration (MIC90) | > 250 μM | |||
| Administration Time | 24 h | ||||
| Evaluation Method | Antifungal activity of the tested compounds against yeast cells was determined in 96-well plates, with minor modifications (CLSI, 2008). Suspensions of the microorganisms were prepared by taking one loop of pure culture into sterile water and adjusting optical density to 0.1 at 660 nm wavelength and further 50-fold dilution in RPMI 1640 medium, resulting in cell concentration of approximately 2 104 CFU/m. Then, 100 μL of cells were added to the wells of a 96-well microtiter plate that contained 16-500 μM tested compounds and 16-500 μM of FLC or 0.2-62 μM AmB. The plates were incubated for 24 h at 37 °C. Each test was performed in triplicate. The final concentration of DMSO was ensured to be around 1% in all experiments. The MIC90 was defined as the lowest concentration of drug showing no visible growth; MIC50 was defined as the concentration that yielded at least 50% growth inhibition when compared with the growth control well. For determination of minimum fungicidal concentrations (MFC), small aliquots of suspensions (around 10 μL) from each well were transferred using the pipette to YPD agar plates without inhibitors and incubated for 24 h at 37 °C. The MFC was defined as the lowest concentration of drug at which no growth of the colonies was observed. | ||||
| MOA of PDC |
Herein we report the chemical synthesis and biological activity of conjugates of fluconazole with (i) cell-penetrating peptides (CPP), namely TP10-NH2 and TP10-7-NH2, or (ii) antimicrobial peptides (AMP), such as LFcinB(2-11)-NH2, LFcinB[Nle1,11]-NH2 and HLopt2-NH2. Both constituents of produced conjugates (FLC and peptides) display different modes of antifungal activity and affect different molecular targets within fungal cells. CPPs serving as carrier peptides and are considered the fundamental part of an extensively developed concept of a drug delivery system; TP-10 is one of the most promising peptides in this family. These peptides effectively penetrate cell membrane. In our recent paper. we showed that TP10-NH2 can be successfully applied to design conjugates with antimicrobial activity. Modified fragments of bovine lactoferrin (LFcin) and human lactoferrin (HLopt2) are members of the AMP family, peptides that constitute an important segment of the bodys natural immunity against microorganisms.
Click to Show/Hide
|
||||
| Description |
The inactivity of FLCpOH confirmed that the drug derivative, used during the synthesis of our target compounds, did not exert any antifungal activity. All compounds displayed poor activity against the C. glabrata strain tested. Another identified advantage of FLCpOH-TP10-NH2 and FLCpOH-TP10-7-NH2 is their higher fungicidal potential compared to fluconazole. The MFC values against C. albicans reference strains were the same or only twice higher compared to MIC90. The high fungicidal activity of both conjugates was also confirmed with the kill-time assay. However, at least a 4 x MFC concentration was necessary to obtain an evident killing effect after 24 h of treatment.
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| In Vitro Model | Breast cancer | MCF-7/6 cell | CVCL_W972 | ||
| Experiment 25 Reporting the Activity Data of This PDC | [1] | ||||
| Indication | <i>Candida spp</i> infection | ||||
| Efficacy Data | Half minimum inhibitory concentration (MIC90) | > 250 μM | |||
| Administration Time | 24 h | ||||
| Evaluation Method | Antifungal activity of the tested compounds against yeast cells was determined in 96-well plates, with minor modifications (CLSI, 2008). Suspensions of the microorganisms were prepared by taking one loop of pure culture into sterile water and adjusting optical density to 0.1 at 660 nm wavelength and further 50-fold dilution in RPMI 1640 medium, resulting in cell concentration of approximately 2 104 CFU/m. Then, 100 μL of cells were added to the wells of a 96-well microtiter plate that contained 16-500 μM tested compounds and 16-500 μM of FLC or 0.2-62 μM AmB. The plates were incubated for 24 h at 37 °C. Each test was performed in triplicate. The final concentration of DMSO was ensured to be around 1% in all experiments. The MIC90 was defined as the lowest concentration of drug showing no visible growth; MIC50 was defined as the concentration that yielded at least 50% growth inhibition when compared with the growth control well. For determination of minimum fungicidal concentrations (MFC), small aliquots of suspensions (around 10 μL) from each well were transferred using the pipette to YPD agar plates without inhibitors and incubated for 24 h at 37 °C. The MFC was defined as the lowest concentration of drug at which no growth of the colonies was observed. | ||||
| MOA of PDC |
Herein we report the chemical synthesis and biological activity of conjugates of fluconazole with (i) cell-penetrating peptides (CPP), namely TP10-NH2 and TP10-7-NH2, or (ii) antimicrobial peptides (AMP), such as LFcinB(2-11)-NH2, LFcinB[Nle1,11]-NH2 and HLopt2-NH2. Both constituents of produced conjugates (FLC and peptides) display different modes of antifungal activity and affect different molecular targets within fungal cells. CPPs serving as carrier peptides and are considered the fundamental part of an extensively developed concept of a drug delivery system; TP-10 is one of the most promising peptides in this family. These peptides effectively penetrate cell membrane. In our recent paper. we showed that TP10-NH2 can be successfully applied to design conjugates with antimicrobial activity. Modified fragments of bovine lactoferrin (LFcin) and human lactoferrin (HLopt2) are members of the AMP family, peptides that constitute an important segment of the bodys natural immunity against microorganisms.
Click to Show/Hide
|
||||
| Description |
The inactivity of FLCpOH confirmed that the drug derivative, used during the synthesis of our target compounds, did not exert any antifungal activity. All compounds displayed poor activity against the C. glabrata strain tested. Another identified advantage of FLCpOH-TP10-NH2 and FLCpOH-TP10-7-NH2 is their higher fungicidal potential compared to fluconazole. The MFC values against C. albicans reference strains were the same or only twice higher compared to MIC90. The high fungicidal activity of both conjugates was also confirmed with the kill-time assay. However, at least a 4 x MFC concentration was necessary to obtain an evident killing effect after 24 h of treatment.
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| In Vitro Model | Invasive breast carcinoma | MCF-7/ADR cell | CVCL_0031 | ||
| Experiment 26 Reporting the Activity Data of This PDC | [1] | ||||
| Indication | <i>Candida spp</i> infection | ||||
| Efficacy Data | Half minimum inhibitory concentration (MIC90) | > 250 μM | |||
| Administration Time | 24 h | ||||
| Evaluation Method | Antifungal activity of the tested compounds against yeast cells was determined in 96-well plates, with minor modifications (CLSI, 2008). Suspensions of the microorganisms were prepared by taking one loop of pure culture into sterile water and adjusting optical density to 0.1 at 660 nm wavelength and further 50-fold dilution in RPMI 1640 medium, resulting in cell concentration of approximately 2 104 CFU/m. Then, 100 μL of cells were added to the wells of a 96-well microtiter plate that contained 16-500 μM tested compounds and 16-500 μM of FLC or 0.2-62 μM AmB. The plates were incubated for 24 h at 37 °C. Each test was performed in triplicate. The final concentration of DMSO was ensured to be around 1% in all experiments. The MIC90 was defined as the lowest concentration of drug showing no visible growth; MIC50 was defined as the concentration that yielded at least 50% growth inhibition when compared with the growth control well. For determination of minimum fungicidal concentrations (MFC), small aliquots of suspensions (around 10 μL) from each well were transferred using the pipette to YPD agar plates without inhibitors and incubated for 24 h at 37 °C. The MFC was defined as the lowest concentration of drug at which no growth of the colonies was observed. | ||||
| MOA of PDC |
Herein we report the chemical synthesis and biological activity of conjugates of fluconazole with (i) cell-penetrating peptides (CPP), namely TP10-NH2 and TP10-7-NH2, or (ii) antimicrobial peptides (AMP), such as LFcinB(2-11)-NH2, LFcinB[Nle1,11]-NH2 and HLopt2-NH2. Both constituents of produced conjugates (FLC and peptides) display different modes of antifungal activity and affect different molecular targets within fungal cells. CPPs serving as carrier peptides and are considered the fundamental part of an extensively developed concept of a drug delivery system; TP-10 is one of the most promising peptides in this family. These peptides effectively penetrate cell membrane. In our recent paper. we showed that TP10-NH2 can be successfully applied to design conjugates with antimicrobial activity. Modified fragments of bovine lactoferrin (LFcin) and human lactoferrin (HLopt2) are members of the AMP family, peptides that constitute an important segment of the bodys natural immunity against microorganisms.
Click to Show/Hide
|
||||
| Description |
The inactivity of FLCpOH confirmed that the drug derivative, used during the synthesis of our target compounds, did not exert any antifungal activity. All compounds displayed poor activity against the C. glabrata strain tested. Another identified advantage of FLCpOH-TP10-NH2 and FLCpOH-TP10-7-NH2 is their higher fungicidal potential compared to fluconazole. The MFC values against C. albicans reference strains were the same or only twice higher compared to MIC90. The high fungicidal activity of both conjugates was also confirmed with the kill-time assay. However, at least a 4 x MFC concentration was necessary to obtain an evident killing effect after 24 h of treatment.
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| In Vitro Model | Invasive ductal carcinoma | MCF7/PTX cell | CVCL_C5RS | ||
| Experiment 27 Reporting the Activity Data of This PDC | [1] | ||||
| Indication | <i>Candida spp</i> infection | ||||
| Efficacy Data | Half minimum inhibitory concentration (MIC90) | > 250 μM | |||
| Administration Time | 24 h | ||||
| Evaluation Method | Antifungal activity of the tested compounds against yeast cells was determined in 96-well plates, with minor modifications (CLSI, 2008). Suspensions of the microorganisms were prepared by taking one loop of pure culture into sterile water and adjusting optical density to 0.1 at 660 nm wavelength and further 50-fold dilution in RPMI 1640 medium, resulting in cell concentration of approximately 2 104 CFU/m. Then, 100 μL of cells were added to the wells of a 96-well microtiter plate that contained 16-500 μM tested compounds and 16-500 μM of FLC or 0.2-62 μM AmB. The plates were incubated for 24 h at 37 °C. Each test was performed in triplicate. The final concentration of DMSO was ensured to be around 1% in all experiments. The MIC90 was defined as the lowest concentration of drug showing no visible growth; MIC50 was defined as the concentration that yielded at least 50% growth inhibition when compared with the growth control well. For determination of minimum fungicidal concentrations (MFC), small aliquots of suspensions (around 10 μL) from each well were transferred using the pipette to YPD agar plates without inhibitors and incubated for 24 h at 37 °C. The MFC was defined as the lowest concentration of drug at which no growth of the colonies was observed. | ||||
| MOA of PDC |
Herein we report the chemical synthesis and biological activity of conjugates of fluconazole with (i) cell-penetrating peptides (CPP), namely TP10-NH2 and TP10-7-NH2, or (ii) antimicrobial peptides (AMP), such as LFcinB(2-11)-NH2, LFcinB[Nle1,11]-NH2 and HLopt2-NH2. Both constituents of produced conjugates (FLC and peptides) display different modes of antifungal activity and affect different molecular targets within fungal cells. CPPs serving as carrier peptides and are considered the fundamental part of an extensively developed concept of a drug delivery system; TP-10 is one of the most promising peptides in this family. These peptides effectively penetrate cell membrane. In our recent paper. we showed that TP10-NH2 can be successfully applied to design conjugates with antimicrobial activity. Modified fragments of bovine lactoferrin (LFcin) and human lactoferrin (HLopt2) are members of the AMP family, peptides that constitute an important segment of the bodys natural immunity against microorganisms.
Click to Show/Hide
|
||||
| Description |
The inactivity of FLCpOH confirmed that the drug derivative, used during the synthesis of our target compounds, did not exert any antifungal activity. All compounds displayed poor activity against the C. glabrata strain tested. Another identified advantage of FLCpOH-TP10-NH2 and FLCpOH-TP10-7-NH2 is their higher fungicidal potential compared to fluconazole. The MFC values against C. albicans reference strains were the same or only twice higher compared to MIC90. The high fungicidal activity of both conjugates was also confirmed with the kill-time assay. However, at least a 4 x MFC concentration was necessary to obtain an evident killing effect after 24 h of treatment.
Click to Show/Hide
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| In Vitro Model | Breast adenocarcinoma | MDA-MB-231 (KRAS G13D) cell | CVCL_0062 | ||
| Experiment 28 Reporting the Activity Data of This PDC | [1] | ||||
| Indication | <i>Candida spp</i> infection | ||||
| Efficacy Data | Half minimum inhibitory concentration (MIC90) | > 250 μM | |||
| Administration Time | 24 h | ||||
| Evaluation Method | Antifungal activity of the tested compounds against yeast cells was determined in 96-well plates, with minor modifications (CLSI, 2008). Suspensions of the microorganisms were prepared by taking one loop of pure culture into sterile water and adjusting optical density to 0.1 at 660 nm wavelength and further 50-fold dilution in RPMI 1640 medium, resulting in cell concentration of approximately 2 104 CFU/m. Then, 100 μL of cells were added to the wells of a 96-well microtiter plate that contained 16-500 μM tested compounds and 16-500 μM of FLC or 0.2-62 μM AmB. The plates were incubated for 24 h at 37 °C. Each test was performed in triplicate. The final concentration of DMSO was ensured to be around 1% in all experiments. The MIC90 was defined as the lowest concentration of drug showing no visible growth; MIC50 was defined as the concentration that yielded at least 50% growth inhibition when compared with the growth control well. For determination of minimum fungicidal concentrations (MFC), small aliquots of suspensions (around 10 μL) from each well were transferred using the pipette to YPD agar plates without inhibitors and incubated for 24 h at 37 °C. The MFC was defined as the lowest concentration of drug at which no growth of the colonies was observed. | ||||
| MOA of PDC |
Herein we report the chemical synthesis and biological activity of conjugates of fluconazole with (i) cell-penetrating peptides (CPP), namely TP10-NH2 and TP10-7-NH2, or (ii) antimicrobial peptides (AMP), such as LFcinB(2-11)-NH2, LFcinB[Nle1,11]-NH2 and HLopt2-NH2. Both constituents of produced conjugates (FLC and peptides) display different modes of antifungal activity and affect different molecular targets within fungal cells. CPPs serving as carrier peptides and are considered the fundamental part of an extensively developed concept of a drug delivery system; TP-10 is one of the most promising peptides in this family. These peptides effectively penetrate cell membrane. In our recent paper. we showed that TP10-NH2 can be successfully applied to design conjugates with antimicrobial activity. Modified fragments of bovine lactoferrin (LFcin) and human lactoferrin (HLopt2) are members of the AMP family, peptides that constitute an important segment of the bodys natural immunity against microorganisms.
Click to Show/Hide
|
||||
| Description |
The inactivity of FLCpOH confirmed that the drug derivative, used during the synthesis of our target compounds, did not exert any antifungal activity. All compounds displayed poor activity against the C. glabrata strain tested. Another identified advantage of FLCpOH-TP10-NH2 and FLCpOH-TP10-7-NH2 is their higher fungicidal potential compared to fluconazole. The MFC values against C. albicans reference strains were the same or only twice higher compared to MIC90. The high fungicidal activity of both conjugates was also confirmed with the kill-time assay. However, at least a 4 x MFC concentration was necessary to obtain an evident killing effect after 24 h of treatment.
Click to Show/Hide
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| In Vitro Model | Uterine sarcoma | MES-SA/Dx5 cell | CVCL_2598 | ||
| Experiment 29 Reporting the Activity Data of This PDC | [1] | ||||
| Indication | <i>Candida spp</i> infection | ||||
| Efficacy Data | Half maximal inhibitory concentration (IC50) | > 100 μM | |||
| Administration Time | 24 h | ||||
| Evaluation Method | MTT assay | ||||
| MOA of PDC |
Herein we report the chemical synthesis and biological activity of conjugates of fluconazole with (i) cell-penetrating peptides (CPP), namely TP10-NH2 and TP10-7-NH2, or (ii) antimicrobial peptides (AMP), such as LFcinB(2-11)-NH2, LFcinB[Nle1,11]-NH2 and HLopt2-NH2. Both constituents of produced conjugates (FLC and peptides) display different modes of antifungal activity and affect different molecular targets within fungal cells. CPPs serving as carrier peptides and are considered the fundamental part of an extensively developed concept of a drug delivery system; TP-10 is one of the most promising peptides in this family. These peptides effectively penetrate cell membrane. In our recent paper. we showed that TP10-NH2 can be successfully applied to design conjugates with antimicrobial activity. Modified fragments of bovine lactoferrin (LFcin) and human lactoferrin (HLopt2) are members of the AMP family, peptides that constitute an important segment of the bodys natural immunity against microorganisms.
Click to Show/Hide
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| Description |
The adverse drug effects associated with the use of antimicrobials can be a major concern especially with antifungal agents due to the eukaryotic nature of both the organism being targeted and the host. Therefore, it was important to test the toxicity of novel fluconazole-based conjugates. To examine and compare the cytotoxicity of FLC and its derivatives, the cytotoxicities of the compounds were assessed by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay in two mammalian cell lines: human foreskin fibroblast cell line (Hs27) and human umbilical venous endothelial primary cells (HUVEC). The results showed that the FLC appeared to be less cytotoxic than its conjugates (Table 3). Among conjugates, FLCpOH-TP10-NH2 and FLCpOH-TP10-7-NH2 appeared to be more cytotoxic. However, the IC90 value for the human cells after 72 h treatment was comparable to the MIC50 value after 24 h treatment for most strains of C. albicans (Table S2).
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| In Vitro Model | Mycobacterium abscessus infection | Mycobacterium abscessus | 36809 | ||
| Experiment 30 Reporting the Activity Data of This PDC | [1] | ||||
| Indication | <i>Candida spp</i> infection | ||||
| Efficacy Data | Half maximal inhibitory concentration (IC50) | > 100 μM | |||
| Administration Time | 24 h | ||||
| Evaluation Method | MTT assay | ||||
| MOA of PDC |
Herein we report the chemical synthesis and biological activity of conjugates of fluconazole with (i) cell-penetrating peptides (CPP), namely TP10-NH2 and TP10-7-NH2, or (ii) antimicrobial peptides (AMP), such as LFcinB(2-11)-NH2, LFcinB[Nle1,11]-NH2 and HLopt2-NH2. Both constituents of produced conjugates (FLC and peptides) display different modes of antifungal activity and affect different molecular targets within fungal cells. CPPs serving as carrier peptides and are considered the fundamental part of an extensively developed concept of a drug delivery system; TP-10 is one of the most promising peptides in this family. These peptides effectively penetrate cell membrane. In our recent paper. we showed that TP10-NH2 can be successfully applied to design conjugates with antimicrobial activity. Modified fragments of bovine lactoferrin (LFcin) and human lactoferrin (HLopt2) are members of the AMP family, peptides that constitute an important segment of the bodys natural immunity against microorganisms.
Click to Show/Hide
|
||||
| Description |
The adverse drug effects associated with the use of antimicrobials can be a major concern especially with antifungal agents due to the eukaryotic nature of both the organism being targeted and the host. Therefore, it was important to test the toxicity of novel fluconazole-based conjugates. To examine and compare the cytotoxicity of FLC and its derivatives, the cytotoxicities of the compounds were assessed by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay in two mammalian cell lines: human foreskin fibroblast cell line (Hs27) and human umbilical venous endothelial primary cells (HUVEC). The results showed that the FLC appeared to be less cytotoxic than its conjugates (Table 3). Among conjugates, FLCpOH-TP10-NH2 and FLCpOH-TP10-7-NH2 appeared to be more cytotoxic. However, the IC90 value for the human cells after 72 h treatment was comparable to the MIC50 value after 24 h treatment for most strains of C. albicans (Table S2).
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| In Vitro Model | Normal | Human umbilical vein endothelial cells | Homo sapiens | ||
| Experiment 31 Reporting the Activity Data of This PDC | [1] | ||||
| Indication | <i>Candida spp</i> infection | ||||
| Efficacy Data | 90% maximal inhibitory concentration (IC50) | > 100 μM | |||
| Administration Time | 24 h | ||||
| Evaluation Method | MTT assay | ||||
| MOA of PDC |
Herein we report the chemical synthesis and biological activity of conjugates of fluconazole with (i) cell-penetrating peptides (CPP), namely TP10-NH2 and TP10-7-NH2, or (ii) antimicrobial peptides (AMP), such as LFcinB(2-11)-NH2, LFcinB[Nle1,11]-NH2 and HLopt2-NH2. Both constituents of produced conjugates (FLC and peptides) display different modes of antifungal activity and affect different molecular targets within fungal cells. CPPs serving as carrier peptides and are considered the fundamental part of an extensively developed concept of a drug delivery system; TP-10 is one of the most promising peptides in this family. These peptides effectively penetrate cell membrane. In our recent paper. we showed that TP10-NH2 can be successfully applied to design conjugates with antimicrobial activity. Modified fragments of bovine lactoferrin (LFcin) and human lactoferrin (HLopt2) are members of the AMP family, peptides that constitute an important segment of the bodys natural immunity against microorganisms.
Click to Show/Hide
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| Description |
The adverse drug effects associated with the use of antimicrobials can be a major concern especially with antifungal agents due to the eukaryotic nature of both the organism being targeted and the host. Therefore, it was important to test the toxicity of novel fluconazole-based conjugates. To examine and compare the cytotoxicity of FLC and its derivatives, the cytotoxicities of the compounds were assessed by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay in two mammalian cell lines: human foreskin fibroblast cell line (Hs27) and human umbilical venous endothelial primary cells (HUVEC). The results showed that the FLC appeared to be less cytotoxic than its conjugates (Table 3). Among conjugates, FLCpOH-TP10-NH2 and FLCpOH-TP10-7-NH2 appeared to be more cytotoxic. However, the IC90 value for the human cells after 72 h treatment was comparable to the MIC50 value after 24 h treatment for most strains of C. albicans (Table S2).
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| In Vitro Model | Mycobacterium abscessus infection | Mycobacterium abscessus | 36809 | ||
| Experiment 32 Reporting the Activity Data of This PDC | [1] | ||||
| Indication | <i>Candida spp</i> infection | ||||
| Efficacy Data | 90% maximal inhibitory concentration (IC50) | > 100 μM | |||
| Administration Time | 24 h | ||||
| Evaluation Method | MTT assay | ||||
| MOA of PDC |
Herein we report the chemical synthesis and biological activity of conjugates of fluconazole with (i) cell-penetrating peptides (CPP), namely TP10-NH2 and TP10-7-NH2, or (ii) antimicrobial peptides (AMP), such as LFcinB(2-11)-NH2, LFcinB[Nle1,11]-NH2 and HLopt2-NH2. Both constituents of produced conjugates (FLC and peptides) display different modes of antifungal activity and affect different molecular targets within fungal cells. CPPs serving as carrier peptides and are considered the fundamental part of an extensively developed concept of a drug delivery system; TP-10 is one of the most promising peptides in this family. These peptides effectively penetrate cell membrane. In our recent paper. we showed that TP10-NH2 can be successfully applied to design conjugates with antimicrobial activity. Modified fragments of bovine lactoferrin (LFcin) and human lactoferrin (HLopt2) are members of the AMP family, peptides that constitute an important segment of the bodys natural immunity against microorganisms.
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| Description |
The adverse drug effects associated with the use of antimicrobials can be a major concern especially with antifungal agents due to the eukaryotic nature of both the organism being targeted and the host. Therefore, it was important to test the toxicity of novel fluconazole-based conjugates. To examine and compare the cytotoxicity of FLC and its derivatives, the cytotoxicities of the compounds were assessed by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay in two mammalian cell lines: human foreskin fibroblast cell line (Hs27) and human umbilical venous endothelial primary cells (HUVEC). The results showed that the FLC appeared to be less cytotoxic than its conjugates (Table 3). Among conjugates, FLCpOH-TP10-NH2 and FLCpOH-TP10-7-NH2 appeared to be more cytotoxic. However, the IC90 value for the human cells after 72 h treatment was comparable to the MIC50 value after 24 h treatment for most strains of C. albicans (Table S2).
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| In Vitro Model | Normal | Human umbilical vein endothelial cells | Homo sapiens | ||
References