| Size | Price | Stock | Qty |
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| 50mg |
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| 100mg |
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| Other Sizes |
Purity: ≥98%
| Targets |
S. pneumoniae[1] IC50: 25.75 nM (Plasmodium falciparum)[3] ggTas2r1, ggTas2r2 and ggTas2r7[4]
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| ln Vitro |
Using fluctuation analysis, the mutation rate to ethylenedimine (optochin) resistance is calculated in three capsulated S. strains of pneumoniae (S. pneumoniae R6 ATCC BAA-255, S. pneumoniae D39 NCTC 7466, and S. pneumoniae ATCC 49619). For all three strains under investigation, exposure to subinhibitory amounts of penicillin elevated the mutation rate (measured as mutation per cell division) to ethylhydrocupreine (Optochin) resistance by 2.1–3.1 fold[2].
Ethylhydrocupreine hydrochloride shows bactericidal activity against Type I pneumococci in pleural pus: equal parts of pus and 1:1,000 solution at 37°C achieve complete sterilization within 30 min; final dilution as high as 1:4,000 is bactericidal. No appreciable effect on Staphylococcus aureus at 1:200 dilution after 2 h exposure. [1] Against Plasmodium falciparum chloroquine-susceptible clone 3D7, Ethylhydrocupreine exhibited an IC50 of 25.75 nM (0.02575 μM) in a [3H]hypoxanthine incorporation assay. [3] In HEK293T cells expressing chicken bitter taste receptors, ethylhydrocupreine activated ggTas2r1, ggTas2r2, and ggTas2r7 with threshold concentrations around 10 μM as measured by calcium mobilization. [4] Optochin at 6 mg/L is used in BHI agar plates to select for optochin-resistant mutants of Streptococcus pneumoniae in fluctuation analysis. [2] |
| ln Vivo |
Acute suppurative pleuritis on both sides is associated with suppurative pericarditis when 1 cc of a 24-hour dextrose blood broth culture of virulent Type I pneumococci is injected into the right pleural cavity of guinea pigs. One milliliter (1:500) of ethylhydrocupreine hydrochloride injections, given to each guinea pig's pleural cavity at different intervals for up to 24 hours following pleural infection, have typically demonstrated a significant curative effect. Dogs exhibit comparable outcomes [1].
In rabbit pneumococcus meningitis models, subthecal injection of 0.0005 g/kg ethylhydrocupreine hydrochloride within 4-6 h after infection had a beneficial effect. [1] In guinea pig pneumococcus pleuritis (induced by intrapleural injection of 1 cc Type I pneumococcus culture), intrapleural injection of 1 cc of 1:500 solution (≈2.5 cc/kg) into each pleural sac immediately or within 24 h after infection markedly reduced inflammation and prolonged survival; animals treated within 24 h often lived without signs of illness, while untreated controls died within 48 h with bilateral suppurative pleuritis and bacteremia. Treatment delayed to 48 h was ineffective. [1] In dogs (5 kg) with pneumococcus pleuritis, daily intrapleural injections of 15 cc of 1:500 solution (≈0.03 g/kg) starting 24 h after infection (6 doses) resulted in survival with sterile pleural sacs and normal lungs; untreated control died at 72 h. [1] |
| Cell Assay |
Bacterial fluctuation analysis (optochin resistance assay): Streptococcus pneumoniae strains were grown in brain heart infusion (BHI) broth with or without subinhibitory antibiotic concentrations. Cultures (0.4 mL) were spread onto BHI agar plates containing 6 mg/L optochin. Mutation rate (mutations per cell division) was estimated using fluctuation analysis with at least 20 replicas, and the FT program was used for statistical analysis. [2]
Calcium mobilization assay for chicken Tas2rs: HEK293T cells stably expressing chimeric G protein Gα16gust44 were transiently transfected with ggTas2r1, ggTas2r2, or ggTas2r7 constructs. About 24 h post-transfection, cells were loaded with the calcium-sensitive dye Fluo-4 AM in the presence of 2.5 mM probenecid, washed, and placed in a fluorometric imaging plate reader (FLIPR). Various concentrations of bitter compounds were automatically applied, and changes in fluorescence were monitored. A second application of 100 nM SST-14 (somatostatin) was included as a vitality control. [4] In vitro antimalarial assay: The chloroquine-susceptible P. falciparum clone 3D7 was cultured in a modified Trager-Jensen method. Drugs were preloaded in 96-well plates at nine concentrations (7.62×10⁻⁴ to 5 μM). Ring-stage parasitized erythrocytes (0.5% parasitaemia, 1.8% haematocrit) were added (200 μL/well). After 48 h, [³H]hypoxanthine was added and plates incubated for another 24 h. Parasites were harvested and radioactivity measured by liquid scintillation counting. IC50 was estimated by linear regression of log10 concentration versus inhibition percentage. [3] |
| Animal Protocol |
Pneumococcus pleuritis in guinea pigs: Guinea pigs weighing 350-500 g were infected by injecting 1 cc of a 24-hour dextrose broth culture of virulent Type I pneumococci into the right pleural sac using aseptic technique. Ethylhydrocupreine hydrochloride was prepared as a 1:500 dilution in sterile saline, warmed to 37°C, and slowly injected into the right pleural sac (single dose) or both pleural sacs (1 cc each) at intervals ranging from immediately to 48 h post-infection. Dose equivalent to ~2.5 cc per kg (0.005 g per kg) per sac. Multiple dose experiments: daily injections of 0.5 cc culture into right sac; treatment started 4 h after second culture injection, with 0.5 cc of 1:500 drug into each sac repeated daily 4 h after each culture injection. Controls received culture alone or drug alone. Necropsies were performed aseptically for films and cultures of heart blood and exudates; sections of pleura and lungs were prepared. [1]
Pneumococcus pleuritis in dogs: Dogs weighing ~5 kg were infected by injecting 15 cc of a 24-hour broth culture reinforced with cocci from ten blood agar slants into the right pleural cavity. Treatment began 24 h after infection with 15 cc of warm 1:500 ethylhydrocupreine hydrochloride injected into the right pleural cavity, repeated daily for 4-7 doses. [1] Toxicity study in guinea pigs: Intrapleural injection of 1:100 solution (0.5-1 cc per 100 g) of ethylhydrocupreine hydrochloride into guinea pigs weighing 350-400 g. Solutions were warmed and slowly injected. Highest tolerated dose was 0.2-0.3 cc of 1:100 solution per 100 g (equivalent to 2-3 cc or 0.02-0.03 g per kg). [1] Toxicity study in dogs: Slow injection of up to 15 cc per kg of warm 1:500 solution (0.030 g per kg) into the right pleural cavity of normal dogs caused no immediate symptoms except occasional short coughs; necropsy 24-72 h later showed small amount of fluid and slight hyperemic changes in lung. [1] |
| Toxicity/Toxicokinetics |
In guinea pigs (intrapleural route): Highest tolerated dose of ethylhydrocupreine hydrochloride is 0.2-0.3 cc of 1:100 solution per 100 g body weight (equivalent to 0.02-0.03 g per kg). Toxic doses caused convulsions, coughing, and death within minutes to hours. Intrapleural toxicity was slightly higher than intravenous. Doses 2.5-5 times lower than toxic (e.g., 1 cc of 1:500 per 350-500 g guinea pig) produced no gross or histological pleural irritation. [1]
In dogs (intrapleural): 15 cc per kg of 1:500 solution (0.030 g per kg) produced no discernible toxicity but caused slight hyperemia and leucocytic infiltration of pleura on histological examination. [1] Systemic toxicity in humans (oral/intramuscular): Transient symptoms include tinnitus, deafness, amblyopia, or retinitis; retinitis or amaurosis may result in permanent vision impairment. [1] |
| References |
[1]. J A Kolmer, et al. CHEMOTHERAPEUTIC STUDIES WITH ETHYLHYDROCUPREINE HYDROCHLORIDE IN EXPERIMENTAL PNEUMOCOCCUS PLEURITIS. J Exp Med. 1921 May 31;33(6):693-711.
[2]. Paulo R Cortes, et al. Subinhibitory Concentrations of Penicillin Increase the Mutation Rate to Optochin Resistance in Streptococcus Pneumoniae. J Antimicrob Chemother. 2008 Nov;62(5):973-7. [3]. Nassira Mahmoudi, et al. Identification of New Antimalarial Drugs by Linear Discriminant Analysis and Topological Virtual Screening. J Antimicrob Chemother. 2006 Mar;57(3):489-97. [4]. Antonella Di Pizio, et al. Molecular Features Underlying Selectivity in Chicken Bitter Taste Receptors. Front Mol Biosci. 2018 Jan 31;5:6. |
| Additional Infomation |
Ethylhydrocupreine hydrochloride is also known as Optochin. It exhibits high selective bactericidal activity for pneumococci in vitro, even in the presence of proteins. However, systemic administration (oral, intramuscular, intravenous) is too toxic to achieve therapeutic concentrations. Direct injection into serous cavities (subarachnoid, pleural) allows locally high drug concentrations with minimal systemic toxicity. The drug was used experimentally for pneumococcal meningitis and pleuritis. [1]
In Streptococcus pneumoniae, resistance to optochin is conferred by point mutations in the atpAC genes encoding subunits of F₀F₁-ATPase; optochin susceptibility testing (optochin disc) is used for clinical identification of pneumococci. [2] Ethylhydrocupreine was identified as a potential antimalarial drug through topological virtual screening and showed in vitro activity against P. falciparum. [3] In chickens, ethylhydrocupreine activates all three bitter taste receptors (ggTas2r1, ggTas2r2, ggTas2r7) with distinct selectivity profiles; it is more selective for ggTas2r1 than for ggTas2r2 and ggTas2r7. [4] |
| Molecular Formula |
C21H29CLN2O2
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|---|---|
| Molecular Weight |
376.92
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| Exact Mass |
376.192
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| CAS # |
3413-58-9
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| Related CAS # |
Ethylhydrocupreine;522-60-1
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| PubChem CID |
16219340
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| Appearance |
Typically exists as solid at room temperature
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| Boiling Point |
508.7ºC at 760 mmHg
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| Melting Point |
121-123ºC
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| Flash Point |
261.5ºC
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| LogP |
4.527
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| Hydrogen Bond Donor Count |
2
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| Hydrogen Bond Acceptor Count |
4
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| Rotatable Bond Count |
5
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| Heavy Atom Count |
26
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| Complexity |
446
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| Defined Atom Stereocenter Count |
4
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| SMILES |
CC[C@H]1CN2CC[C@H]1C[C@H]2[C@@H](C3=C4C=C(C=CC4=NC=C3)OCC)O.Cl
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| InChi Key |
QNRATNLHPGXHMA-XZHTYLCXSA-N
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| InChi Code |
InChI=1S/C21H28N2O2.ClH/c1-3-14-13-23-10-8-15(14)11-20(23)21(24)17-7-9-22-19-6-5-16(25-4-2)12-18(17)19;/h5-7,9,12,14-15,20-21,24H,3-4,8,10-11,13H2,1-2H3;1H/t14-,15-,20-,21+;/m0./s1
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| Chemical Name |
(R)-(6-ethoxyquinolin-4-yl)-[(2S,4S,5R)-5-ethyl-1-azabicyclo[2.2.2]octan-2-yl]methanol;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 (e.g. under nitrogen), avoid exposure to moisture and light. |
| 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 : 125 mg/mL (331.64 mM)
H2O : 50 mg/mL (132.65 mM) |
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.08 mg/mL (5.52 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 20.8 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: ≥ 2.08 mg/mL (5.52 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (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 20.8 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly. View More
Solubility in Formulation 3: 33.33 mg/mL (88.43 mM) in PBS (add these co-solvents sequentially from left to right, and one by one), clear solution; with ultrasonication. |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.6531 mL | 13.2654 mL | 26.5308 mL | |
| 5 mM | 0.5306 mL | 2.6531 mL | 5.3062 mL | |
| 10 mM | 0.2653 mL | 1.3265 mL | 2.6531 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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