Baloxavir (Xofluza; BXA; S033447)

Alias: Trade name Xofluza; Baloxavir acid; BXA; Baloxavir marboxil; S-033447; S 033447; S033447; Baloxavir;
Cat No.:V0094 Purity: ≥98%
Baloxavir(trade name Xofluza;also known asBaloxaviracid, BXA,or S-033447),derived from the prodrug baloxavir marboxil (BXM), is an orally available small molecule inhibitor of the cap-dependent endonuclease.
Baloxavir (Xofluza; BXA; S033447) Chemical Structure CAS No.: 1985605-59-1
Product category: Influenza Virus
This product is for research use only, not for human use. We do not sell to patients.
Size Price Stock Qty
500μg
1mg
5mg
10mg
25mg
50mg
100mg
Other Sizes

Other Forms of Baloxavir (Xofluza; BXA; S033447):

  • Baloxavir marboxil (BXM, S033188)
  • Baloxavir-d4 (Baloxavir acid-d4; S-033447-d4)
Official Supplier of:
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Purity & Quality Control Documentation

Purity: ≥98%

Product Description

Baloxavir (trade name Xofluza; also known as Baloxavir acid, BXA,or S-033447), derived from the prodrug baloxavir marboxil (BXM), is an orally available small molecule inhibitor of the cap-dependent endonuclease. It is an antiviral drug developed by Roche and Shionogi as an anti-influenza agent for treatment of influenza A and influenza B flu. As of 2018, it was approved for clinical use in Japan and in the United States. Baloxavir was discovered by rational molecular design based on the two-metal pharmacophore concept for dolutegravir (DTG), a strand transfer inhibitor of human immunodeficiency virus (HIV) integrase. Baloxavir potently and selectively inhibits the cap-dependent endonuclease within the polymerase PA subunit of influenza A and B viruses. In February 2018, baloxavir received its first global approval in Japan for the treatment of influenza A or B virus infections. Phase III development is underway in the USA, EU and other countries for this indication.The drug blocks influenza virus proliferation by inhibiting the initiation of mRNA synthesis. In clinical trials, single doses of Baloxavir profoundly decrease viral titers as well as alleviating influenza symptoms.

Biological Activity I Assay Protocols (From Reference)
Targets
Influenza virus[1] Cap-dependent endonuclease (CEN)[1][2]
ln Vitro
For A/H1N1pdm, A/H3N2, and type B virus, the median EC50 values at baseline for baloxavir (BXA) are 17.96 nM, 4.48 nM, and 18.67 nM, respectively[1].
In enzymatic assays, baloxavir (BXA) selectively inhibits cap-dependent endonuclease (CEN) activity to prevent viral RNA transcription. In cytopathic effect assays, BXA prevents viral replication in infected cells without causing cytotoxicity. When it comes to different subtypes of influenza A viruses, baloxavir exhibits broad potency (H1N2, H5N1, H5N2, H5N6, H7N9, and H9N2). Additionally, isolates PA/I38T variants with lower susceptibility to BXA are obtained through serial passages of the viruses in the presence of baloxavir[2].
Baloxavir (BXA) inhibits cap-dependent endonuclease (CEN) and CEN/RdRp activities with IC50 values of 2.5 nM and 1.6 nM, respectively, while low potency (IC50 >40 nM) is observed against RdRp activity[2].
Baloxavir (BXA) has a high inhibitory potency against CEN activity of the tested viral ribonucleoprotein complexes (vRNPs) from influenza A and B viruses with mean IC50 values of 1.4-3.1 nM and 4.5-8.9 nM, respectively, indicating that Baloxavir has broad spectrum activities. Baloxavir shows high potency against influenza A and B viruses with mean EC90 of 0.46 - 0.98 nM and 2.2-3.4 nM, respectively[2].
ln Vivo
In clinical trials, single doses of Baloxavir profoundly decrease viral titers as well as alleviating influenza symptoms.
Enzyme Assay
Oseltamivir acid was serially diluted in MES assay buffer [32.5 mmol/L MES and 4 mmol/L CaCl2 in DW (pH 6.5 adjusted with 4 N NaOH)]. To prepare NA enzyme solution, virus stocks were inactivated by 0.1% NP-40, and diluted with MES assay buffer. Ten μL of the oseltamivir acid solution and 10 μL of the NA enzyme solution were mixed and incubated at 37 °C for 30 minutes, followed by addition of 30 μL of 100 μmol/L 2′-(4-Methylumbelliferyl)-α-D-N-acetylneuraminic acid sodium salt hydrate (MUNANA; Sigma-Aldrich Co., Ltd.). The reaction mixtures were incubated at 37 °C for 60 minutes, and the reaction was stopped by addition of 150 μL of stop solution [0.1 mol/L glycine and 25% ethanol (pH 10.7 adjusted with 4 N NaOH)]. The fluorescence intensity was measured with a microplate reader EnVision 2103 (PerkinElmer Inc.) at excitation wavelength of 355 nm and an emission wavelength of 460 nm, followed by calculation of IC50 values with XLfit software. FC was calculated by dividing IC50 of each tested virus to IC50 of the cognate wild-type virus.
Cell Assay
Canine kidney MDCK cells were obtained from European Collection of Cell Cultures. Human quasi-diploid tumor RPMI2650 and human embryonic kidney 293 T cells were provided by American Type Culture Collection. MDCK and RPMI2650 cells were maintained in minimal essential medium (MEM) supplemented with 10% fetal bovine serum (FBS) and 100 µg/mL kanamycin (Thermo Fisher Scientific, Inc.). 293 T cells were cultured in Dulbecco’s modified Eagle’s medium with 10% FBS and 100 µg/mL kanamycin. Eight plasmids-based reverse genetics technique was employed to generate recombinant viruses as described. The plasmid set of rgA/WSN/33 (H1N1) and empty vector pHW2000 were provided by Dr. Robert Webster at St. Jude Children’s Research Hospital. The plasmids for the generation of rgA/Victoria/3/75 and rgB/Maryland viruses were constructed with the pHW2000 by standard molecular biology techniques. The primer sequences used are available upon request. Co-culture of MDCK and 293 T cells were transfected with the eight plasmids and incubated 48 to 72 hours, followed by propagation of the viruses in MDCK cells. The PA sequences of the recombinant viruses were verified by Sanger sequencing. Viral titers were determined by standard tissue culture infectious dose (TCID)50 assay or plaque-forming unit (PFU) assay in MDCK cells.
Animal Protocol
N/A
N/A
References

[1]. Sci Rep. 2018 Jun 25;8(1):9633.

[2]. Antiviral Res. 2018 Dec;160:109-117.

[3].Sci Rep.2018 Jun 25;8(1):9633

[4].Drugs.2018 Apr;78(6):693-697

[5].WO 2017104691 A1.

These protocols are for reference only. InvivoChem does not independently validate these methods.
Physicochemical Properties
Molecular Formula
C₂₄H₁₉F₂N₃O₄S
Molecular Weight
483.49
Exact Mass
483.11
Elemental Analysis
C, 59.62; H, 3.96; F, 7.86; N, 8.69; O, 13.24; S, 6.63
CAS #
1985605-59-1
Appearance
Solid powder
SMILES
O=C1N2[C@](COCC2)([H])N([C@@H]3C4=CC=CC=C4SCC5=C(F)C(F)=CC=C35)N6C1=C(O)C(C=C6)=O
InChi Key
FIDLLEYNNRGVFR-CTNGQTDRSA-N
InChi Code
InChI=1S/C24H19F2N3O4S/c25-16-6-5-13-15(20(16)26)12-34-18-4-2-1-3-14(18)21(13)29-19-11-33-10-9-27(19)24(32)22-23(31)17(30)7-8-28(22)29/h1-8,19,21,31H,9-12H2/t19-,21+/m1/s1
Chemical Name
({(12aR)-12-[(11S)-7,8-difluoro-6,11-dihydrodibenzo[b,e]thiepin-11-yl]-6,8-dioxo-3,4,6,8,12,12ahexahydro-1H-[1,4]oxazino[3,4-c]pyrido[2,1-f][1,2,4]triazin-7-yl}oxy)
Synonyms
Trade name Xofluza; Baloxavir acid; BXA; Baloxavir marboxil; S-033447; S 033447; S033447; Baloxavir;
HS Tariff Code
2934.99.9001
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)
Solubility Data
Solubility (In Vitro)
DMSO : 25~41.67 mg/mL ( 51.7~86.19 mM )
Solubility (In Vivo)
10% DMSO+40% PEG300+5% Tween-80+45% Saline : ≥ 2.08 mg/mL (4.30 mM) (Please use freshly prepared in vivo formulations for optimal results.)
Preparing Stock Solutions 1 mg 5 mg 10 mg
1 mM 2.0683 mL 10.3415 mL 20.6830 mL
5 mM 0.4137 mL 2.0683 mL 4.1366 mL
10 mM 0.2068 mL 1.0341 mL 2.0683 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.

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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.
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Biological Data
  • Baloxavir


    In vitroendonuclease activity and inhibition of PA variants and thermal stabilization induced by the binding of BXA.2018 Jun 25;8(1):9633.

  • Baloxavir


    BXA binding to influenza A/H1N1 PA endonuclease. BXA interacts with (A) PA-A WT and (B) PA-A I38T by chelating the two manganese ions in the active site.2018 Jun 25;8(1):9633.

  • Baloxavir


    Comparison of PA endonuclease from Flu A and Flu B bound to BXA in either WT or I38T form. Superposition of PA-BXA complexes: (A) PA-A WT and PA-A I38T, (B) PA-B WT and PA-B I38T, (C) PA-A WT and PA-B WT, (D) PA-A I38T and PA-B I38T.2018 Jun 25;8(1):9633.

  • Baloxavir


    BXA binding to influenza B/Memphis PA endonuclease As Fig.4, but for (A) PA-B WT and (B) PA-B I38T.2018 Jun 25;8(1):9633.

  • Baloxavir


    Local interactions of residue 38 in apo- and BXA-bound FluB PA (A) Superposition of ligand-free PA-B WT (PDB:5FML, in hotpink) and bound to BXA (green sticks for BXA, teal sticks/cartoon for PA). (B) Superposition of ligand-free (forest green) and BXA-bound PA-B I38T (light magenta sticks for BXA, orange sticks/cartoon for PA).2018 Jun 25;8(1):9633.

  • Baloxavir


    Replicative capacity of variant viruses with indicated AA substitutions in PA protein. Canine MDCK cells (A–C) or human RPMI2650 cells (D,E) were infected with WT or I38x viruses based on rgA/WSN/33 (H1N1) (A,D), rgA/Victoria/3/75 (H3N2) (B,E), or B/Maryland/1/5

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