| Size | Price | Stock | Qty |
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| 5mg |
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| 25mg |
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| 50mg |
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| Targets |
GSK369796 Dihydrochloride targets ferriprotoporphyrin IX (FP, heme) in the malaria parasite, inhibiting hemozoin crystallization. Equilibrium binding constants (log K1 and log K2) for heme binding in 40% DMSO: log K1 = 5.23 (±0.1), log K2 = 5.36 (±0.1), best-fit model 2 (stepwise addition of two heme molecules to one drug) [1].
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| ln Vitro |
GSK369796 DiHClide has an IC50 of 11.2±2.2, 12.6±5.3, and 13.2±3.2 nM, respectively, which inhibits the development of Plasmodium falciparum strains 3D7c, HB3c, and K1d in vitro. In mice, GSK369796 dihydrochloride (chemical 4) exhibited stronger protein binding (93% vs.74%) than desethylamodiaquine, although human protein binding (88% vs.86%) was comparable. GSK369796 DiHClide has an IC50 of 7.5±0.8 μM and can also prevent hERG potassium channel repolarization[1].
GSK369796 Dihydrochloride showed potent in vitro antimalarial activity against Plasmodium falciparum strains: IC50 (nM) vs 3D7 (chloroquine-sensitive) = 11.2 ± 2.2; vs HB3 (chloroquine-sensitive) = 12.6 ± 5.3; vs K1 (chloroquine-resistant) = 13.2 ± 3.2. In comparison, chloroquine IC50 vs K1 = 183.2 ± 11.1 nM, amodiaquine = 15.5 ± 9.4 nM [1]. Against field isolates from patients failing amodiaquine treatment (Thailand, Kenya, Rwanda), GSK369796 Dihydrochloride was superior to chloroquine and desethyl amodiaquine in vitro [1]. Mechanism of action: UV-visible spectroscopic binding studies with heme in 40% DMSO revealed that the binding stoichiometry involves addition of two heme molecules per drug, similar to chloroquine and amodiaquine. Molecular modeling showed the quinoline ring parallel to the heme edge (π-π stacking) and hydrogen bonding between heme carboxylates and the Mannich side chain alcohol/protonated amine [1]. |
| ln Vivo |
Plasmodium berghei ANKA is inhibited in vivo by GSK369796 DiHClide, with ED50 and ED90 values of 2.8 and 4.7 mg/kg, respectively [1].
GSK369796 Dihydrochloride displayed oral efficacy against Plasmodium berghei ANKA in mice (standard 4-day test): ED50 = 2.8 mg/kg (95% CI 2.3–3.3), ED90 = 4.7 mg/kg (95% CI 4.1–5.5). Against Plasmodium yoelii 17X: ED50 = 3.8 mg/kg (95% CI 3.3–4.3), ED90 = 5.4 mg/kg (95% CI 4.8–6.1), non-recrudescence level (NRL) = 20 mg/kg. Comparable activity to amodiaquine but with better antimalarial exposure profile [1]. In infected mice (10% P. yoelii parasitemia), after single 10 mg/kg oral administration, blood exposure profiles were substantially higher than in non-infected animals (see Figure 5) [1]. |
| Enzyme Assay |
Heme binding assay: UV-visible spectroscopic method in mixed aqueous/organic solvent (buffered 40% DMSO) to provide strictly monomeric heme species. Titrations were carried out, and absorbance spectra were recorded after each addition. Two binding models were mathematically fitted (stepwise binding of two drug equivalents to one heme, or stepwise addition of two hemes to one drug). Data were analyzed using Pro-Fit nonlinear curve fitting to obtain K1 and K2 values at χ² minimum, initially at one wavelength then simultaneously at 10–15 wavelengths [1].
Cytochrome P450 inhibition assay: Potential for inhibition of major human CYP isozymes (1A2, 2C8, 2C9, 2C19, 2D6, 3A4) was determined using recombinant isozymes and fluorescent probe substrates. IC50 values (mean ± SD, n=3) were calculated. For GSK369796 Dihydrochloride: CYP1A2 IC50 = 29 ± 6 μM; CYP2D6 = 3.0 ± 0.5 μM; CYP2C8 = 23 ± 2 μM; CYP2C9 ≥90 μM; CYP2C19 = 24 ± 10 μM; CYP3A4 (DEF) >100 μM; CYP3A4 (PPR) activation observed (3-fold stimulation) [1]. |
| Cell Assay |
GSK369796 Dihydrochloride was tested against P. falciparum cultures in a 48 h [3H]-hypoxanthine incorporation assay. Parasite cultures (2% parasitemia, 0.5% hematocrit) in RPMI 1640 with 25 mM HEPES, 32 mM NaHCO3, and 10% human serum were incubated with drug dilutions for 24 h, then 0.5 μCi hypoxanthine added for another 24 h. Harvested onto filter mats, dried, and counted by liquid scintillation. IC50 calculated by probit transformation of log dose-response curve [1].
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| Animal Protocol |
Mouse efficacy model (P. berghei ANKA or P. yoelii 17X): Female CD1 mice (n=5/group) were infected intravenously with 6.4×10⁶ or 10.0×10⁶ parasites on day 0. Treatments (in sterile saline) were administered orally once daily from day 0 (1 h post-infection) to day 3. On day 4, parasitemia measured by flow cytometry using YOYO-1 staining. Recrudescence monitored up to day 22–25 if parasitemia <0.01% at day 4 [1].
PK studies: Single intravenous and oral administration to mouse, rat, dog, and/or monkey. Blood clearance (CL), oral bioavailability (F), volume of distribution, and half-life determined. Protein binding and blood cell association measured in plasma or whole blood from preclinical species and humans in vitro using equilibrium dialysis or rapid equilibrium dialysis (RED). Metabolite profiling in liver microsomes and hepatocytes from mouse, rat, dog, and human. Urine collected from mice after oral administration of 4 or 5 (10 mg/kg) for metabolite identification by LC-MS/MS [1]. |
| ADME/Pharmacokinetics |
Blood clearance (CL) of GSK369796 Dihydrochloride in animals (mL/min/kg): mouse = 17, rat = 26 ± 5, dog = 6.3 ± 1.6, monkey = 14.5 ± 0.7. Oral bioavailability (F%): mouse ~100, rat 89 ± 12, dog 68 ± 18, monkey ~100. Steady-state distribution volume high, exceeding total body water, indicating extensive tissue penetration. Elimination half-life: ~3 h in mouse, ~11 h in monkey. Blood-to-plasma ratio: human ~1.2, dog ~15. Protein binding: mouse 93%, human 88%. In infected mice, blood exposure higher than in non-infected [1].
Metabolism: GSK369796 Dihydrochloride is metabolized via mono-oxygenation (M2, M3, M4), aldehyde (M5), carboxylic acid (M6), glucuronides of parent/M5/M6, and a ketone metabolite (M7). N-dealkylation (M1) was not detected in vitro but was detected in mouse urine (very low levels compared to desethyl isoquine). No glutathione conjugation or reactive quinoneimine metabolites observed [1]. |
| Toxicity/Toxicokinetics |
hERG inhibition: GSK369796 Dihydrochloride inhibited cloned hERG potassium channels with IC50 = 7.5 ± 0.8 μM (chloroquine IC50 = 2.5 μM, amodiaquine IC50 = 2.4 ± 0.3 μM). In monkeys, mild transient increases in QTc intervals and some ECG rhythm abnormalities were observed [1].
Genetic toxicology: GSK369796 Dihydrochloride did not cause gene mutations or chromosomal damage in bacterial mutation assay (Ames), mouse lymphoma assay, or in vivo mouse micronucleus assay [1]. CNS effects: Adverse dose-related CNS effects (respiratory depression, tremors, convulsions) observed in preclinical safety pharmacology and toxicity studies, similar to chloroquine. Off-target receptor profiling (CEREP screen) showed antagonistic activity at muscarinic and 5HT receptors [1]. Cardiovascular toxicity: In rat 4–14 day studies, high doses of GSK369796 Dihydrochloride, chloroquine, and amodiaquine caused myocardial necrosis, skeletal muscle myopathy, hepatic effects (hypertrophy, transaminase release, apoptosis). GSK369796 Dihydrochloride appeared safer than chloroquine and no worse than amodiaquine. Short-term (4-day) dosing at tolerated doses did not show cardiac effects with 4, whereas chloroquine did [1]. Phospholipidosis: Macroscopic and microscopic evidence of phospholipidosis in multiple tissues (reversible, not affecting organ function) at exposures comparable to predicted human efficacy [1]. |
| References | |
| Additional Infomation |
GSK369796 Dihydrochloride (N-tert-butyl isoquine) was selected based on superior activity against chloroquine-resistant parasites, improved metabolic stability (reduced N-dealkylation compared to isoquine), and elimination of the para-aminophenol metabolic alert present in amodiaquine. The optimized two-step synthesis yields >98% purity with cost of goods equivalent or less than chloroquine. The molecule meets the target product profile for an oral antimalarial set by MMV and WHO. Preclinical toxicology revealed liabilities that did not translate into human experience, highlighting limitations of current preclinical testing for this class. Predicted human plasma Cmax = 0.1–0.3 μg/mL, AUC = 16 μg·h/mL. The drug is intended for short-term (3-day) dosing [1].
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| Molecular Formula |
C20H24CL3N3O
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|---|---|
| Molecular Weight |
428.7831
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| Exact Mass |
427.098
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| CAS # |
1010411-21-8
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| Related CAS # |
459133-38-1;1010411-21-8 (HCl);
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| PubChem CID |
51071988
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| Appearance |
Light yellow to yellow solid powder
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| Hydrogen Bond Donor Count |
5
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| Hydrogen Bond Acceptor Count |
4
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| Rotatable Bond Count |
5
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| Heavy Atom Count |
27
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| Complexity |
428
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| Defined Atom Stereocenter Count |
0
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| InChi Key |
UDVALKJFXQVZSI-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C20H22ClN3O.2ClH/c1-20(2,3)23-12-13-4-6-15(11-19(13)25)24-17-8-9-22-18-10-14(21)5-7-16(17)18;;/h4-11,23,25H,12H2,1-3H3,(H,22,24);2*1H
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| Chemical Name |
2-[(tert-Butylamino)methyl]-5-[(7-chloroquinolin-4-yl)amino]phenol Dihydrochloride
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| Synonyms |
GSK369796 dihydrochloride; GSK 369796; GSK-369796; GSK369796; N-tert-Butyl isoquine.
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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 : ~100 mg/mL (~233.22 mM)
H2O : ~50 mg/mL (~116.61 mM) |
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (5.83 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 (5.83 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 | 2.3322 mL | 11.6610 mL | 23.3220 mL | |
| 5 mM | 0.4664 mL | 2.3322 mL | 4.6644 mL | |
| 10 mM | 0.2332 mL | 1.1661 mL | 2.3322 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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