| Size | Price | Stock | Qty |
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| 1mg |
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| Other Sizes |
| Targets |
Cycloguanil acts as a potent inhibitor of dihydrofolate reductase (DHFR), an enzyme critical for folate metabolism and DNA synthesis. It binds to the DHFR active site with high affinity: Ki values are 1.5 nM for Plasmodium falciparum DHFR and 0.79 nM for Plasmodium berghei DHFR. This inhibition depletes tetrahydrofolate, blocking thymidylate and DNA synthesis. In human cancer cells, it inhibits human DHFR with an IC50 of approximately 10.8 microM and blocks STAT3 transcriptional activity. The d6-labeled analog retains equivalent binding properties but is used quantitatively.
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| ln Vitro |
The unlabeled parent compound cycloguanil shows potent in vitro antimalarial activity against Plasmodium falciparum strains, with IC50 values in the low nanomolar range (e.g., 1-20 nM). It also exhibits antiproliferative activity against various human cancer cell lines (e.g., leukemia, colon, breast) via DHFR inhibition. The compound acts synergistically with sulfonamides or sulfones, which inhibit the subsequent dihydropteroate synthase, in the classical antifolate combination therapy. However, cycloguanil-d6 is not used for functional assays.
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| ln Vivo |
In vivo, the unlabeled parent cycloguanil demonstrates efficacy in murine malaria models (e.g., P. berghei-infected mice) when administered via oral or intraperitoneal routes. The ED50 values are typically in the range of 1-5 mg/kg. The drug is also evaluated in tumor-bearing mouse models (cancer research) due to its DHFR inhibitory activity. However, the major clinical use of the parent proguanil is in combination with atovaquone for malaria prophylaxis and treatment. Cycloguanil-d6 is strictly an analytical standard and not administered to animals for efficacy assessment.
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| Enzyme Assay |
Cycloguanil-d6 is not used in standard receptor binding assays as it is an internal standard. Its identity as a labeled compound is confirmed by NMR and mass spectrometry. For LC-MS/MS method validation, a stock solution is prepared at a concentration of 1 mg/mL in DMSO or methanol. The exact level of deuterium enrichment (typically >98%) is determined by mass spectrometry. The compound is characterized by the molecular formula C11H₈D₆ClN₅ (if the free base) or as the hydrochloride salt.
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| Cell Assay |
For analytical quantitation of cycloguanil, a standard operating procedure (SOP) using Cycloguanil-d6 as internal standard: 1) To 100 microL of plasma, add 10 microL of a 100 ng/mL solution of Cycloguanil-d6 internal standard. 2) Perform protein precipitation by adding 300 microL of ice-cold acetonitrile, vortex for 1 minute, and centrifuge at 14,000 rpm for 10 minutes. 3) Transfer the supernatant to a clean vial and evaporate to dryness under nitrogen. 4) Reconstitute in 100 microL of mobile phase (80:20, acetonitrile:0.1% formic acid). 5) Inject 10 microL onto an LC-MS/MS system (C18 column, 2.6 microm, 2.1×50 mm) with a total run time of 3-5 minutes. Monitor MRM transitions: parent drug and d6-labeled compound.
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| Animal Protocol |
Cycloguanil-d6 is not applied to cell culture because it serves as an analytical reference standard. For the evaluation of antimalarial activity of unlabeled cycloguanil in vitro, a standard protocol is: P. falciparum parasites (3D7 strain, chloroquine-sensitive) are cultured in human O+ erythrocytes (hematocrit 4%) in RPMI-1640 medium with 25 mM HEPES and 10% human serum. Parasites are synchronized with 5% sorbitol. Ring-stage parasites (0.5% parasitemia, 1% hematocrit) are incubated with serial dilutions of cycloguanil (0.1-1000 nM) in 96-well plates for 48 hours at 37degC under 5% CO2, 5% O2, 90% N2. [3H]hypoxanthine (0.5 microCi/well) is added for the final 24 hours. Plates are harvested onto glass fiber filters, and radioactivity is counted. IC50s are calculated by non-linear regression.
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| ADME/Pharmacokinetics |
The unlabeled cycloguanil is used in animal malaria models. For the P. berghei ANKA strain murine malaria model, female BALB/c or C57BL/6 mice (6-8 weeks, n=5 per group) are infected intraperitoneally with 1 × 10⁶ parasitized erythrocytes. Treatment with cycloguanil (administered orally or i.p. at 1-25 mg/kg) begins 2 hours post-infection and continues once daily for 4 days. Parasitemia is assessed microscopically on Giemsa-stained thin tail blood smears on days 4, 7, and 14. The primary endpoint is suppression of parasitemia on day 4 relative to vehicle controls. ED50 and ED90 values are calculated. Survival is monitored up to day 30. Cycloguanil-d6 is used to analyze drug levels in blood and organs in separate pharmacokinetic cohorts.
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| Toxicity/Toxicokinetics |
The unlabeled cycloguanil is rapidly absorbed after oral administration of proguanil (which is metabolized to cycloguanil by CYP2C19 and other enzymes). In humans, peak plasma concentrations of cycloguanil are achieved 2-5 hours after proguanil administration. The elimination half-life is approximately 12-20 hours. Cycloguanil binds moderately to plasma proteins (approx. 60-75%) and is primarily excreted renally (as unchanged drug and metabolites). Deuterium labeling (d6) slows in vivo metabolism to a minor extent in some contexts, enhancing the stability of the internal standard during bioanalysis.
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| Additional Infomation |
Cycloguanil-d6 is a stable isotope-labeled reagent used at trace concentrations (ng/mL); it does not present acute toxicity hazards under normal laboratory handling. The unlabeled parent drug cycloguanil, as an antimalarial dihydrofolate reductase inhibitor, has a well-characterized safety profile. Common adverse effects at therapeutic doses include mild gastrointestinal disturbances (nausea, abdominal pain, diarrhea) and mouth ulcers. At high doses, cycloguanil can cause reversible bone marrow suppression (leukopenia, thrombocytopenia) due to folate antagonism, as well as hepatocellular injury. In animal models, no significant carcinogenicity or genotoxicity has been reported for the parent drug, but teratogenicity is a potential risk (folate antagonist). Cycloguanil-d6 is for research use only.
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| Molecular Formula |
C11H8D6CLN5
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| Molecular Weight |
257.75
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| Related CAS # |
Cycloguanil;516-21-2;Cycloguanil-d6 nitrate
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| Appearance |
White to off-white solid powder
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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 |
| 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) |
May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples
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| Solubility (In Vivo) |
Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.
Injection Formulations
Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)(e.g. IP/IV/IM/SC) *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution. Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil) Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals). View More
Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)] Oral Formulations
Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium) Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals). View More
Oral Formulation 3: Dissolved in PEG400 (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 3.8797 mL | 19.3986 mL | 38.7973 mL | |
| 5 mM | 0.7759 mL | 3.8797 mL | 7.7595 mL | |
| 10 mM | 0.3880 mL | 1.9399 mL | 3.8797 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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