| Size | Price | Stock | Qty |
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| 100mg |
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| 250mg |
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| Targets |
Trypanothione reductase (TR) is the primary molecular target of Antitrypanosomal agent 1. TR is a flavoprotein oxidoreductase that is unique to trypanosomatid parasites and is essential for maintaining intracellular redox balance by reducing trypanothione disulfide (T[S]₂) to trypanothione (T[SH]₂) using NADPH as a cofactor. This enzyme plays a central role in the parasite's defense against oxidative stress, which is critical for survival within the host. The compound acts as an antagonist, binding to the enzyme and inhibiting its catalytic activity. The inhibition is competitive with respect to the substrate trypanothione. Antitrypanosomal agent 1 also inhibits glutathione reductase (GR), a mammalian homolog, but with an IC₅₀ of 64.8 μM, which is approximately 20-fold higher than its IC₅₀ for TR, demonstrating significant selectivity for the parasitic enzyme. This selectivity is a key feature of the compound, as it suggests that the compound can effectively target the parasite's redox machinery while sparing the host's equivalent system. The inhibition of TR leads to a disruption of the parasite's thiol redox balance, accumulation of trypanothione disulfide, and increased susceptibility to oxidative damage, ultimately resulting in parasite death.
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| ln Vitro |
Antitrypanosoma agent 1 (compound 8a) has a selectivity of 20 for TR (GR IC50/TR IC50) [1].
In cell-free enzymatic assays, Antitrypanosomal agent 1 demonstrates potent and specific inhibition of trypanothione reductase (TR) with an IC₅₀ of 3.3 μM. This activity was determined by measuring the decrease in absorbance at 340 nm, which corresponds to the oxidation of NADPH, in a reaction mixture containing recombinant TR, its substrate trypanothione disulfide, and varying concentrations of the compound. The compound's inhibitory effect on the mammalian homolog glutathione reductase (GR) was significantly weaker, with an IC₅₀ of 64.8 μM, yielding a selectivity index of approximately 20 for TR over GR. This selectivity profile indicates that the compound preferentially targets the parasitic enzyme, potentially reducing the risk of host toxicity. The compound's in vitro activity against the parasite itself was confirmed in cellular assays using Trypanosoma brucei, where it exhibited an EC₅₀ of 1 μM. The correlation between enzyme inhibition and antiparasitic activity suggests that the primary mechanism of action is indeed the inhibition of TR. The compound's activity is dose-dependent, and its efficacy is likely influenced by factors such as cellular uptake and intracellular accumulation. |
| ln Vivo |
In vivo activity data for Antitrypanosomal agent 1 is limited in the available literature. However, based on its potent in vitro activity against Trypanosoma brucei (EC₅₀ = 1 μM) and its favorable physicochemical properties, including a molecular weight of 282.59 and a logP of 3.929, it is expected to exhibit some degree of in vivo efficacy in animal models of African trypanosomiasis. Typically, such studies involve a murine model where mice are infected with Trypanosoma brucei and the compound is administered via oral gavage or intraperitoneal injection at various doses. Efficacy is assessed by monitoring parasitemia through microscopic examination of blood smears and recording survival time. The compound's moderate lipophilicity suggests it may have reasonable oral bioavailability and tissue distribution, which are important for reaching parasites in the bloodstream and other tissues. However, detailed in vivo pharmacokinetic and pharmacodynamic studies, including dose-response relationships and efficacy against different stages of the disease, are necessary to fully evaluate its therapeutic potential. The compound's selectivity for TR over GR suggests a favorable safety profile, but this needs to be confirmed in vivo.
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| Enzyme Assay |
The standard protocol for evaluating the inhibition of trypanothione reductase (TR) in a cell-free system involves a spectrophotometric assay. Recombinant TR is expressed and purified from a suitable expression system. The assay mixture typically contains 100 mM potassium phosphate buffer (pH 7.5), 0.2 mM NADPH, 0.2 mM trypanothione disulfide, and the test compound at varying concentrations. The reaction is initiated by the addition of the enzyme, and the decrease in absorbance at 340 nm, which is proportional to the oxidation of NADPH, is monitored over time using a microplate reader. The initial velocity of the reaction is calculated from the linear portion of the absorbance curve. The percentage of inhibition at each compound concentration is calculated relative to a control without inhibitor. The IC₅₀ value, representing the concentration of compound required to inhibit 50% of the enzyme activity, is determined by fitting the data to a four-parameter logistic model. To assess selectivity, the same protocol is applied using glutathione reductase (GR) from a mammalian source, with glutathione disulfide as the substrate. The compound's inhibition of GR is compared to its inhibition of TR to calculate the selectivity index.
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| Cell Assay |
The in vitro cellular activity of Antitrypanosomal agent 1 is typically assessed using a resazurin-based fluorescence assay. Trypanosoma brucei parasites are cultured in appropriate medium, such as HMI-9 supplemented with fetal bovine serum, at 37°C in a humidified atmosphere containing 5% CO₂. The parasites are seeded in 96-well plates at a density of approximately 2 × 10⁴ cells per well. Serial dilutions of the test compound are added to the wells, and the plates are incubated for 72 hours. After incubation, resazurin (a non-fluorescent blue dye) is added to each well. Viable cells with active metabolism reduce resazurin to resorufin, a highly fluorescent pink compound. The fluorescence intensity is measured using a microplate reader with excitation at 560 nm and emission at 590 nm. The fluorescence signal is directly proportional to the number of viable cells. The percentage of inhibition at each compound concentration is calculated relative to untreated control wells. The EC₅₀ value, representing the concentration that reduces parasite viability by 50%, is determined from the dose-response curve. Cytotoxicity against mammalian cells, such as human HeLa or HEK293 cells, is often assessed in parallel using the same assay to determine the selectivity index.
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| Animal Protocol |
For in vivo efficacy studies, a murine model of African trypanosomiasis is commonly employed. Female BALB/c or CD-1 mice, typically 6-8 weeks old, are infected intraperitoneally with approximately 1 × 10⁴ Trypanosoma brucei parasites. The infection is allowed to establish for 1-3 days. Antitrypanosomal agent 1 is then administered to the mice at various doses, typically via oral gavage or intraperitoneal injection, once or twice daily for a period of 4-7 days. A control group receives the vehicle alone. Parasitemia is monitored daily by microscopic examination of tail-vein blood smears. The number of parasites per high-power field is counted, and the level of infection is graded. Body weight and survival are recorded throughout the study. At the end of the treatment period, the efficacy of the compound is evaluated by comparing parasitemia levels and survival rates between treated and control groups. A compound is considered effective if it significantly reduces parasitemia and prolongs survival compared to the vehicle control. For a more rigorous assessment, a "cure" model may be used, where mice are monitored for an extended period after treatment to determine if the infection has been completely cleared.
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| ADME/Pharmacokinetics |
Detailed pharmacokinetic properties for Antitrypanosomal agent 1 are not extensively reported in the available literature. However, based on its physicochemical characteristics, some predictions can be made. The compound has a molecular weight of 282.59, a logP of 3.929, and a moderate number of hydrogen bond donors (1) and acceptors (2). These properties suggest that the compound is likely to have moderate oral bioavailability, as it falls within the Lipinski's Rule of Five parameters for drug-likeness. The compound is soluble in DMSO (~25 mg/mL) and water (~10 mg/mL), indicating reasonable aqueous solubility for formulation. In vivo, the compound is expected to be distributed throughout the body, including the bloodstream and tissues, which is important for treating systemic infections like African trypanosomiasis. Metabolism is likely to occur via hepatic cytochrome P450 enzymes, with elimination occurring through renal or biliary routes. However, specific parameters such as half-life (t₁/₂), volume of distribution (Vd), clearance (CL), and oral bioavailability (%F) have not been determined experimentally. Further studies, including plasma protein binding and metabolic stability assays, are needed to fully characterize the pharmacokinetic profile of this compound.
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| Toxicity/Toxicokinetics |
Toxicological data for Antitrypanosomal agent 1 is limited, as it is a research chemical not intended for human use. Standard safety precautions should be observed when handling this compound. No specific toxicity studies, such as acute or chronic toxicity in animal models, genotoxicity, or cardiotoxicity (e.g., hERG channel inhibition), have been detailed in the public domain. However, the compound's selectivity for trypanothione reductase over glutathione reductase (IC₅₀ of 3.3 μM vs. 64.8 μM) suggests that it may have a favorable safety profile, as it is less likely to inhibit the critical host enzyme. The compound's moderate lipophilicity (logP 3.929) may be associated with some risk of off-target effects, but this has not been assessed. In the absence of formal toxicology studies, researchers should handle the compound with care, using appropriate personal protective equipment and following institutional safety guidelines. The compound is not approved for clinical use and should only be used in preclinical research settings.
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| References |
[1]. Martyn DC, et al. High-throughput screening affords novel and selective trypanothione reductase inhibitors with anti-trypanosomal activity. Bioorg Med Chem Lett. 2007 Mar 1;17(5):1280-3.
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| Additional Infomation |
Antitrypanosomal agent 1 is a research tool for studying trypanothione metabolism and developing new treatments for African trypanosomiasis. Its mechanism of action involves the selective inhibition of trypanothione reductase, a key enzyme in the parasite's antioxidant defense. The compound was identified through high-throughput screening and is cited in medicinal chemistry literature. It is not approved for clinical use.
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| Molecular Formula |
C11H14CL3NO
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| Molecular Weight |
282.5940
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| Exact Mass |
281.014
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| CAS # |
75144-12-6
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| PubChem CID |
2876896
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| Appearance |
White to off-white solid powder
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| Density |
1.22g/cm3
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| Boiling Point |
341.9ºC at 760 mmHg
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| Flash Point |
160.6ºC
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| Index of Refraction |
1.544
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| LogP |
3.929
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| Hydrogen Bond Donor Count |
1
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| Hydrogen Bond Acceptor Count |
2
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| Rotatable Bond Count |
4
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| Heavy Atom Count |
16
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| Complexity |
221
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| Defined Atom Stereocenter Count |
0
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| SMILES |
ClC1=C(C([H])=C([H])C(=C1[H])C(C([H])([H])C([H])([H])N(C([H])([H])[H])C([H])([H])[H])=O)Cl.Cl[H]
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| InChi Key |
PRRKQCKNKRZOPV-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C11H13Cl2NO.ClH/c1-14(2)6-5-11(15)8-3-4-9(12)10(13)7-8;/h3-4,7H,5-6H2,1-2H3;1H
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| Chemical Name |
1-(3,4-dichlorophenyl)-3-(dimethylamino)propan-1-one;hydrochloride
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| HS Tariff Code |
2934.99.9001
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| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month Note: Please store this product in a sealed and protected environment, avoid exposure to moisture. |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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| Solubility (In Vitro) |
DMSO : ~25 mg/mL (~88.47 mM)
H2O : ~10 mg/mL (~35.39 mM) |
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (8.85 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 2: ≥ 2.5 mg/mL (8.85 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 3.5387 mL | 17.6935 mL | 35.3870 mL | |
| 5 mM | 0.7077 mL | 3.5387 mL | 7.0774 mL | |
| 10 mM | 0.3539 mL | 1.7693 mL | 3.5387 mL |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
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