Identification of new drug candidates against Borrelia burgdorferi using high-throughput screening

Venkata Raveendra Pothineni, Dhananjay Wagh, Mustafeez Mujtaba Babar, Mohammed Inayathullah, David Solow-Cordero, Kwang-Min Kim, Aneesh V Samineni, Mansi B Parekh, Lobat Tayebi, Jayakumar Rajadas, Venkata Raveendra Pothineni, Dhananjay Wagh, Mustafeez Mujtaba Babar, Mohammed Inayathullah, David Solow-Cordero, Kwang-Min Kim, Aneesh V Samineni, Mansi B Parekh, Lobat Tayebi, Jayakumar Rajadas

Abstract

Lyme disease is the most common zoonotic bacterial disease in North America. It is estimated that >300,000 cases per annum are reported in USA alone. A total of 10%-20% of patients who have been treated with antibiotic therapy report the recrudescence of symptoms, such as muscle and joint pain, psychosocial and cognitive difficulties, and generalized fatigue. This condition is referred to as posttreatment Lyme disease syndrome. While there is no evidence for the presence of viable infectious organisms in individuals with posttreatment Lyme disease syndrome, some researchers found surviving Borrelia burgdorferi population in rodents and primates even after antibiotic treatment. Although such observations need more ratification, there is unmet need for developing the therapeutic agents that focus on removing the persisting bacterial form of B. burgdorferi in rodent and nonhuman primates. For this purpose, high-throughput screening was done using BacTiter-Glo assay for four compound libraries to identify candidates that stop the growth of B. burgdorferi in vitro. The four chemical libraries containing 4,366 compounds (80% Food and Drug Administration [FDA] approved) that were screened are Library of Pharmacologically Active Compounds (LOPAC1280), the National Institutes of Health Clinical Collection, the Microsource Spectrum, and the Biomol FDA. We subsequently identified 150 unique compounds, which inhibited >90% of B. burgdorferi growth at a concentration of <25 µM. These 150 unique compounds comprise many safe antibiotics, chemical compounds, and also small molecules from plant sources. Of the 150 unique compounds, 101 compounds are FDA approved. We selected the top 20 FDA-approved molecules based on safety and potency and studied their minimum inhibitory concentration and minimum bactericidal concentration. The promising safe FDA-approved candidates that show low minimum inhibitory concentration and minimum bactericidal concentration values can be chosen as lead molecules for further advanced studies.

Keywords: BacTiter-Glo assay; Borrelia burgdorferi; Lyme disease; high-throughput screening; persisters.

Figures

Figure 1
Figure 1
Inhibition assay of drugs on CA8 strain. Notes: Effect of drugs on Borrelia cell viability was studied with drugs: (A) tetraethylthiuram disulfide, doxorubicin hydrochloride, and epirubicin hydrochloride and (B) azlocillin sodium and cephalothin sodium. The control has no drugs. The results represent mean ± SD. Abbreviation: SD, standard deviation.
Figure 2
Figure 2
Time kill curves for B. burgdorferi s.s. isolates CA8 with (A) azlocillin sodium and (B) cefotaxime acid. Notes: The Borrelia was grown in the drug concentrations of 0.625 µM, 1.25 µM, 2.5 µM, and 5 µM. Experiment was performed with triplicates by the investigation of growth using conventional cell counts, and data were reported as the mean of triplicate. The control has no drugs. Abbreviation:B. burgdorferi, Borrelia burgdorferi.

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