| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| 50mg |
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| 100mg |
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| 250mg | |||
| Other Sizes |
| Targets |
Host retrograde trafficking pathway (specific cellular target unknown). The compound inhibits infections by non-enveloped DNA viruses (e.g., JC polyomavirus (JCPyV), human papillomavirus 16 (HPV16)) and toxins (e.g., ricin, Shiga-like toxins) that exploit this pathway for cellular entry.【1】
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| ln Vitro |
Retro-2cycl exhibits antiviral activity against JCPyV and HPV16 in cell-based infection assays. The half-maximal inhibitory concentration (IC50) for inhibiting JCPyV infectivity in SVG-A cells is 54 µM, and for inhibiting HPV16 pseudovirus infectivity in HeLaM cells is 160 µM. Through structural optimization, an analog designated as DHQZ 36 showed significantly improved potency, with IC50 values of 8.1 µM against JCPyV and 24 µM against HPV16, representing a 6.7-fold improvement over the parent compound.【1】
A proximity ligation assay confirmed that Retro-2cycl and its optimized analog DHQZ 36 interfere with retrograde trafficking. Pretreatment of cells with these compounds reduced the co-localization signal between JCPyV capsid protein VP1 and the endoplasmic reticulum (ER) marker PDI, as well as between HPV16 capsid protein L1 and the Golgi marker TGN46, compared to DMSO-treated controls.【1】 |
| Cell Assay |
Inhibition of JCPyV Infection: SVG-A cells were seeded in 12-well plates. After overnight incubation, cells were pre-treated with 25 µM of Retro-2cycl, its analogs, or a DMSO vehicle control for 0.5 hours. JCPyV was then added at a multiplicity of infection (MOI) of 0.25 and infection proceeded for 72 hours in the continued presence of the compound. Cells were then processed for flow cytometry. Infected cells were detected by intracellular staining with an Alexa Fluor-labeled monoclonal antibody against the viral capsid protein VP1 (PAB 597-AF488). Infection rates were normalized to an untreated infected control (~20% infected cells set to 100%), and the reduction in infection in compound-treated samples was calculated relative to this control.【1】
IC50 Determination: Serial dilutions of Retro-2cycl or DHQZ 36 were prepared in DMSO and added to cell culture media. SVG-A cells (for JCPyV) or HeLaM cells (for HPV16 pseudovirus) were pre-treated with the compound dilutions and then challenged with the respective virus as described above. Infection rates were scored by flow cytometry (VP1 staining for JCPyV, HcRed reporter expression for HPV16) after 72 hours. Dose-response curves were generated from three independent experiments to calculate IC50 values.【1】 Proximity Ligation Assay (PLA): SVG-A cells (for JCPyV) or HeLaM cells (for HPV16) were pre-treated with Retro-2cycl, DHQZ 36, or a vehicle control for 0.5 hours. Cells were then inoculated with the respective virus at an MOI of 100 for 1 hour. Unbound virus was washed away, and fresh media containing the compound was added. Cells were incubated for an additional 8 hours (JCPyV) or 16 hours (HPV16), then fixed with paraformaldehyde and permeabilized. Cells were immunostained overnight (JCPyV: anti-VP1 and anti-PDI) or for 2 hours (HPV16: anti-L1 and anti-TGN46) at specified dilutions. The Duolink proximity ligation assay was performed according to the manufacturer's instructions to detect co-localization (within 40 nm) of the target protein pairs, visualized as fluorescent puncta. Cell nuclei were counterstained with DAPI, and images were acquired by confocal microscopy.【1】 |
| References | |
| Additional Infomation |
Retro-2 cycl is a dihydroquinazolinone (DHQZ) compound that has been shown to inhibit host retrotransport. It protects cells from infection by various human polyomaviruses (e.g., JCPyV, BKPyV) and papillomaviruses (e.g., HPV16), as well as protein toxins such as ricin and shiga toxin, all of which enter cells via retrotransport. [1] Its mechanism of action involves disrupting the intracellular transport of viral particles from the endosome through the Golgi apparatus to the endoplasmic reticulum (ER), thereby preventing the virus from reaching the nucleus to initiate replication. [1] In this study, we synthesized and evaluated several Retro-2 cycl analogs and performed structure-activity relationship (SAR) analysis. Key structural modifications that enhance antiviral activity include: replacing 5-methylthiophene with a 5-ethylthiophene heterocycle, replacing benzamide with 4-fluorobenzylamide, and introducing a fluorine atom at the C-6 position of the dihydroquinazolinone benzo[a] ring. [1]
The specific host cell protein target of the Retro-2 ring remains unclear. The structure-activity relationship data from this study are intended to guide the design of future affinity reagents to identify this target. [1] |
| Molecular Formula |
C19H16N2OS
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|---|---|
| Molecular Weight |
320.408143043518
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| Exact Mass |
320.098
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| CAS # |
1429192-00-6
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| PubChem CID |
71716811
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| Appearance |
White to off-white solid powder
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| LogP |
5.03
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| Hydrogen Bond Donor Count |
1
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| Hydrogen Bond Acceptor Count |
3
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| Rotatable Bond Count |
2
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| Heavy Atom Count |
23
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| Complexity |
437
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| Defined Atom Stereocenter Count |
0
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| InChi Key |
BOAPXDSRULBILC-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C19H16N2OS/c1-13-11-12-17(23-13)18-20-16-10-6-5-9-15(16)19(22)21(18)14-7-3-2-4-8-14/h2-12,18,20H,1H3
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| Chemical Name |
2-(5-methylthiophen-2-yl)-3-phenyl-1,2-dihydroquinazolin-4-one
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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) |
DMSO : ≥ 125 mg/mL (~390.13 mM)
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|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.08 mg/mL (6.49 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 (6.49 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. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 3.1210 mL | 15.6050 mL | 31.2100 mL | |
| 5 mM | 0.6242 mL | 3.1210 mL | 6.2420 mL | |
| 10 mM | 0.3121 mL | 1.5605 mL | 3.1210 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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