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Chemical Structure| 22662-39-1 Chemical Structure| 22662-39-1

Structure of Rafoxanide
CAS No.: 22662-39-1

Chemical Structure| 22662-39-1

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Rafoxanide inhibits SPAK and OSR1, is a salicylanilide used as an antiparasitic agent.

Synonyms: NSC 355278; Disalan; EINECS 245-148-9

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Product Citations

Product Citations

Reunanen, Saku ; Ghemtio, Leo ; Patel, Jayendrakumar Z ; Patel, Darshitkumar R ; Airavaara, Kerttu ; Yli-Kauhaluoma, Jari , et al.

Abstract: Parkinson's disease (PD) is a common neurodegenerative disease, typically treated with levodopa, which alleviates the motor symptoms of the disease. However, levodopa metabolism in peripheral tissues hampers its bioavailability and leads to undesired side-effects. Therefore, co-administration of amino acid decarboxylase (AADC) inhibitors is necessary, but still, up to 50% of levodopa may not reach the brain. Recent evidence suggests that gut microbes, especially Enterococcus faecalis, are also able to metabolize levodopa and affect the bioavailability by utilizing microbial tyrosine decarboxylase (TyrDC) enzyme. The main aim of this study was to develop inhibitors targeting gut microbial and host decarboxylation of levodopa. First, a virtual screen of a library of 158,000 compounds against E. faecalis TyrDC was conducted, combining three methods: molecular docking against the E. faecalis TyrDC homology model, structurebased pharmacophore model, and shape similarity searches based on levodopa, (AADC inhibitor) and (S)-α-fluoromethyltyrosine (TyrDC inhibitor). A total of 394 compounds were selected and tested in vitro by using a cell-based E. faecalis assay measuring inhibition of levodopa metabolism. Three most active compounds (49-92% inhibition at 100 µM) sharing a similar scaffold and a set of commercially available and in-house synthesized analogs were then assessed against purified TyrDC and AADC enzymes. The IC50 values for the most potent compounds for TyrDC and AADC inhibition were 23 µM / 144 µM (compound 1), 42 µM / 199 µM (compound 2) and 51 µM / 182 µM (compound 3), respectively. These compounds also displayed cytotoxic effects on HeLa cells and modest activity against E. faecalis at the same concentration range. The core structure of the compounds presented here can serve as a starting point for the development of a new inhibitor class against TyrDC and AADC, and offers a promising avenue for personalized PD treatment, particularly for patients with high levels of gut microbes expressing the levodopa metabolizing TyrDC enzyme.

Keywords: Levodopa ; Parkinson's disease ; Gut microbiome ; Tyrosine decarboxylase

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Product Details of Rafoxanide

CAS No. :22662-39-1
Formula : C19H11Cl2I2NO3
M.W : 626.01
SMILES Code : O=C(NC1=CC=C(OC2=CC=C(Cl)C=C2)C(Cl)=C1)C3=CC(I)=CC(I)=C3O
Synonyms :
NSC 355278; Disalan; EINECS 245-148-9
MDL No. :MFCD00682899
InChI Key :NEMNPWINWMHUMR-UHFFFAOYSA-N
Pubchem ID :31475

Safety of Rafoxanide

GHS Pictogram:
Signal Word:Warning
Hazard Statements:H302-H413
Precautionary Statements:P264-P270-P273-P301+P312-P330

Isoform Comparison

Biological Activity

In Vitro:

Cell Line
Concentration Treated Time Description References
A549 cells 25 µM 24 hours Assessed the impact of Rafoxanide on HAdV DNA replication, showing significant reduction in HAdV DNA replication. Sci Rep. 2019 Jan 9;9(1):17
Klebsiella pneumoniae KP-9 1 mg/L 24 hours Evaluate the synergistic effect of Rafoxanide with colistin, results showed that Rafoxanide significantly enhanced the bactericidal effect of colistin against KP-1 and KP-9. Cancers (Basel). 2020 May 21;12(5):1314
Klebsiella pneumoniae KP-1 1 mg/L 24 hours Evaluate the synergistic effect of Rafoxanide with colistin, results showed that Rafoxanide significantly enhanced the bactericidal effect of colistin against KP-1 and KP-9. Cancers (Basel). 2020 May 21;12(5):1314
Klebsiella pneumoniae KP-9 1 mg/L 24 hours Evaluate the synergistic effect of Rafoxanide with colistin, results showed that Rafoxanide significantly enhanced the bactericidal effect of colistin against KP-1 and KP-9. mSphere. 2023 Oct 24;8(5):e0023423
Klebsiella pneumoniae KP-1 1 mg/L 24 hours Evaluate the synergistic effect of Rafoxanide with colistin, results showed that Rafoxanide significantly enhanced the bactericidal effect of colistin against KP-1 and KP-9. mSphere. 2023 Oct 24;8(5):e0023423
BEAS-2B cells 7.5, 15, 30 µM 24, 36, 48 hours To evaluate the effect of Rafoxanide on normal lung cell viability, results showed that Rafoxanide had little cytotoxicity in BEAS-2B cells. Sci Rep. 2023 Jan 13;13(1):693
H1299 cells 7.5, 15, 30 µM 24, 36, 48 hours To evaluate the effect of Rafoxanide on H1299 cell viability, results showed that Rafoxanide significantly inhibited H1299 cell growth and migration. Sci Rep. 2023 Jan 13;13(1):693
A549 cells 7.5, 15, 30 µM 24, 36, 48 hours To evaluate the effect of Rafoxanide on A549 cell viability, results showed that Rafoxanide significantly inhibited A549 cell growth and induced apoptosis and cell cycle arrest. Sci Rep. 2023 Jan 13;13(1):693
293β5 cells 2.5 µM 48 hours Evaluated the anti-HAdV activity of Rafoxanide against HAdV5 and HAdV16, showing 100% inhibition of plaque formation at 2.5 μM. Sci Rep. 2019 Jan 9;9(1):17
MRC-5 cells 25 µM 72 hours Evaluated the inhibitory effect of Rafoxanide on HCMV DNA replication, showing significant reduction in HCMV DNA replication. Sci Rep. 2019 Jan 9;9(1):17

In Vivo:

Species
Animal Model
Administration Dosage Frequency Description References
BALB/c nude mice A549 xenograft model Intraperitoneal injection 15 mg/kg Once daily for 14 days To evaluate the anti-tumor effect of Rafoxanide in vivo, results showed that Rafoxanide significantly inhibited the growth of A549 xenograft tumors without severe side effects. Sci Rep. 2023 Jan 13;13(1):693
Galleria mellonella Galleria mellonella larvae Intrahemocoel injection and peroral administration 1600-2000 mg/kg Single dose, observed for 120 hours To evaluate the in vivo toxicity of BH77, the results showed an LD50 greater than 1600 mg/kg for intrahemocoel injection and near 1600-2000 mg/kg for peroral administration. Microbiol Spectr. 2023 Jun 15;11(3):e0306422
Zebrafish [HBx,src,p53−/−,RPIA] transgenic fish Oral gavage 7.5 μM Twice a week for one month To evaluate the efficacy of Rafoxanide combined with oligo-fucoidan in treating HCC, results showed the combination significantly reduced the expression of proliferation markers and histopathological features. Int J Mol Sci. 2022 Oct 11;23(20):12100

Protocol

Bio Calculators
Preparing Stock Solutions 1mg 5mg 10mg

1 mM

5 mM

10 mM

1.60mL

0.32mL

0.16mL

7.99mL

1.60mL

0.80mL

15.97mL

3.19mL

1.60mL

Dissolving Methods
Please choose the appropriate dissolution scheme according to your animal administration guide.For the following dissolution schemes, clear stock solution should be prepared according to in vitro experiments, and then cosolvent should be added in turn:

in order to ensure the reliability of the experimental results, the clarified stock solution can be properly preserved according to the storage conditions; The working fluid for in vivo experiment is recommended to be prepared now and used on the same day;

The percentage shown in front of the following solvent refers to the volume ratio of the solvent in the final solution; If precipitation or precipitation occurs in the preparation process, it can be assisted by heating and/or ultrasound.
Protocol 1
Protocol 2

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