| Size | Price | Stock | Qty |
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| 1mg |
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| 5mg |
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| 10mg |
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| Other Sizes |
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| Targets |
Antiviral; Arenavirus envelope glycoprotein
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| ln Vitro |
LHF-535 is a small-molecule viral entry inhibitor that targets the arenavirus envelope glycoprotein (GP).Strong antiviral activity is shown by LHF-535 against a variety of hemorrhagic fever arenaviruses. With an IC50 of 0.1-0.3 nM, LHF-535 inhibits Lassa GP-pseudotyped lentivirus[2].
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| ln Vivo |
LHF-535 (3, 10 or 30 mg/kg; orally; daily; 14 days) significantly lowers viral titers in plasma, spleen, and liver while shielding mice against a deadly Tacaribe virus challenge.Delaying the first dose of LHF-535 (10 mg/kg) by 1, 2, or 3 days after infection also results in an increase in survival, indicating the effectiveness of LHF-535 as a post-exposure therapeutic in mice[2].
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| Enzyme Assay |
Antiviral assays[2]]
Junín virus yield-reduction assay[2] Virus yield reduction (VYR) experiments were conducted to determine sensitivity to LHF-535 in Junín Romero wild-type and vaccine strains. Varying concentrations of LHF-535 were added to test wells containing 70–80% confluent Vero cells just prior to infection at a multiplicity of infection (MOI) of approximately 0.002. Plates were incubated for 3 days, at which time virus-infected plates were frozen and thawed, and culture supernatants were collected for endpoint titration of infectious virus. The samples were plated on Vero cells and visual cytopathic effect was measured on day 10 post-infection. LHF-535 was tested in triplicate against both Candid#1 and the Romero strain. Work with the pathogenic Romero strain of Junín virus was conducted in a BSL-3+ laboratory by vaccinated personnel. |
| Cell Assay |
Vero and 293T cells were obtained from the American Type Culture Collection (ATCC; Manassas, VA). Vero cells were maintained in minimal essential medium (MEM) supplemented with 10% fetal bovine serum (HyClone Thermo Scientific, Logan, UT). 293T cells were maintained in Dulbecco’s modified Eagle medium (DMEM) supplemented with 10% fetal bovine serum, 2 mM L-glutamine, penicillin (100 U/ml), and streptomycin (100 μg/ml). Tacaribe virus strain TRVL 11573 was obtained from ATCC. The Candid#1 vaccine strain of Junín virus was provided by Robert Tesh (World Reference Center for Emerging Viruses and Arboviruses, The University of Texas Medical Branch, Galveston, TX). The Candid#1 virus stock (~108 PFU/ml) was generated from a clarified lysate following one passage in African green monkey kidney cells (BS-C-1 from ATCC) and two passages in Vero cells. The molecular clone of the Romero strain of Junín virus was provided by Slobodan Paessler (University of Texas Medical Branch, Galveston, TX). The virus was rescued in baby hamster kidney fibroblasts (BHK-21 obtained from ATCC) and the stock (~108 PFU/ml) was prepared from a single passage in Vero cells. Work with the pathogenic Romero strain of Junín virus was conducted in a BSL-3+ laboratory by vaccinated personnel.[2]
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| Animal Protocol |
Tacaribe virus in vivo model[2]
AG129 mice are IFN-α/β and–γ receptor-deficient mice. They were a kind gift from Michael Diamond (Washington University in St. Louis). For Tacaribe virus studies, we used mice that were sex- and age-matched (6–8 weeks old). All animal procedures were approved by the Institutional Animal Care and Use Committee (IACUC) and were conducted at Kineta in a BSL-2 facility. Experimental groups were sized (as specified in the figure legends) to allow for statistical analysis, and all animals were included in the analysis. All animal experiments were conducted in a non-blinded fashion. For the LHF-535 dose titration study, mice were sorted into survival and titer arms and challenged by intraperitoneal (i.p.) injection with 200 PFU of Tacaribe virus. In the survival arm, mice were dosed orally with LHF-535 at 3, 10, or 30 mg/kg/day or with vehicle alone for 14 days with the first dose 30 min prior to infection. Micronized LHF-535 was suspended in 0.5% Methocel E15 and 1% Tween 80. The mice were observed for signs of morbidity and mortality. For the titer arm, mice were sacrificed at 7 days post-challenge; plasma, liver, and spleen samples were collected for assaying virus titers. For the delayed treatment studies, AG129 mice were split into five groups with each receiving LHF-535 30 min prior, and 24, 48, and 72 h post infection along with a vehicle control group. All mice were challenged by i.p. injection with 200 PFU of Tacaribe virus and dosing ceased 14 days post-challenge. The mice were observed for signs of morbidity and mortality and were humanely removed from study if there were clinical observations of inactivity, labored breathing, or excessive weight loss (≥20%). For the pathogenesis studies (LD50 determination), AG129 mice were infected with wild-type or mutant Tacaribe virus via i.p. injection using 10-fold serial dilutions of virus. The Reed-Muench method was used for LD50 calculations |
| References |
[1]. Dongcheng Dai, et al. Antiviral drugs for treatment of arenavirus infection. WO2013123215A2.
[2]. A potent Lassa virus antiviral targets an arenavirus virulence determinant. PLoS Pathog. 2018 Dec 21;14(12):e1007439. |
| Molecular Formula |
C27H28N2O2
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|---|---|
| Molecular Weight |
412.52
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| Exact Mass |
412.22
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| Elemental Analysis |
C, 78.61; H, 6.84; N, 6.79; O, 7.76
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| Related CAS # |
LHF-535;1450929-77-7
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| Appearance |
Typically exists as light yellow to yellow solids at room temperature
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| LogP |
5.8
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| tPSA |
47.3Ų
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| InChi Key |
DBNZTRPIBJSUIX-WAYWQWQTSA-N
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| InChi Code |
InChI=1S/C27H28N2O2/c1-19(2)31-24-14-12-23(13-15-24)29-18-28-25-17-21(9-16-26(25)29)6-5-20-7-10-22(11-8-20)27(3,4)30/h5-19,30H,1-4H3/b6-5-
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| Chemical Name |
(E)-2-(4-(2-(1-(4-Isopropoxyphenyl)-1H-benzo[d]imidazol-5-yl)vinyl)phenyl)propan-2-ol
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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 :~60 mg/mL (~145.45 mM)
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (6.06 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 25.0 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 | 2.4241 mL | 12.1206 mL | 24.2412 mL | |
| 5 mM | 0.4848 mL | 2.4241 mL | 4.8482 mL | |
| 10 mM | 0.2424 mL | 1.2121 mL | 2.4241 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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