Fig 1.
Kaplan-Meier Curves Demonstrate the Ability of OXA Derivatives to Kill Adult Schistosomes In Vitro.
A. CIDD-0150610, B. CIDD-0150303. Both compounds kill 100% of S. mansoni, S. haematobium and S. japonicum compared to OXA that kills 90% of S. mansoni in vitro. OXA derivatives were tested against 10 adult male worms per well. All derivatives were solubilized in 100% DMSO, administered at a final concentration of 143 μM per well for 45 minutes, washed 3 timess with media. 45-minute exposure mimics the exposure time in a human (D. Cioli, pers commun.). All screens were performed in experimental and biological triplicate. Survival was plotted as a percentage over time using Prism/Curve Comparison/ Long-rank (Mantel-cox) test. The p-value threshold for each derivative compared to DMSO was <0.001.
Fig 2.
Kaplan-Meier Curves Demonstrate the Effect of Final Concentration Of 71.5 μm Per Well.
A. OXA, B. CIDD-0149830, C. CIDD-0150610, and D. CIDD-0150303. 610 and 303 will kill 100% of S. mansoni, S. haematobium and S. japonicum. 830 will kill 100% of S. mansoni, 60% of S. haematobium and 56.7% of S. japonicum. OXA will kill 40% of S. mansoni OXA and OXA derivatives were tested against 10 adult male worms per well. All derivatives were solubilized in 100% DMSO. The worms were treated for 45 minutes, washed 3 times with media. 45-minute exposure mimics the exposure time in a human All screens were performed in experimental and biological triplicate. Survival was plotted as a percentage over time using Prism/Curve Comparison/ Long-rank (Mantel-cox) test. The p-value threshold for each derivative compared to DMSO was <0.001.
Fig 3.
Kaplan-Meier Curves Demonstrate the Ability of OXA And OXA Derivatives to Kill Both Genders.
A. OXA to kill both genders of S. mansoni B. CIDD-0149830 against both genders of B.1. S. mansoni, B.2. S. haematobium, and B.3. S. japonicum. C. CIDD-0150610 against both genders of C.1. S. mansoni, C.2. S. haematobium, and C.3. S. japonicum. D. CIDD-0150303 against both genders of D.1. S. mansoni, D.2. S. haematobium, and D.3. S. japonicum. E. Percentages of worm killing. OXA and OXA derivatives were tested against 10 adults of single sex- female and male worms and female and male worms in worm pairs per well. 610 and 303 kill 100% of both gender from S. mansoni, S. haematobium and S. japonicum. 830 kills 100% of paired males, and single-sex females and 96.7% of single-sex females. Paired females were less susceptible to 830. OXA demonstrates the expected level of killing against single-sex males and paired females from S. mansoni, the drug is effective 100% against single-sex females and less effective 60% against paired males. All derivatives were solubilized in 100% DMSO and administered at a final concentration of 143 μM per well for 45 minutes, washed 3 times with media. 45-minute exposure mimics the exposure time in a human. All screens were performed in experimental and biological triplicate. Survival was plotted as a percentage over time using Prism/Curve Comparison/ Long-rank (Mantel-cox) test. The p-value threshold for each derivative compared to DMSO was <0.00.
Fig 4.
Effect of OXA Derivatives on A. S. mansoni B. S haematobium C. S. japonicum Infected Animals.
Five mice per group were infected with S. mansoni, 5 hamsters per group were infected with S. haematobium, and 5 hamsters per group were infected with S. japonicum Worms were collected 10 days after treatment with a single dose of 100 mg/kg by oral gavage compared to the untreated control group. Prism/unpaired t test (P<0.05).
Fig 5.
Kaplan-Meier Curves Demonstrate the Ability of OXA Derivatives to Kill All Life Stages of S. mansoni.
A. CIDD-0149830, B. CIDD-0150610, and C. CIDD-0150303 OXA derivatives were tested against cercaria, 3-hour schistosomula, and male worms for 20 dpi, 25 dpi, 28 dpi, 32 dpi and 45 dpi. OXA derivatives demonstrate 100% of killing for all life stages within 2–6 days. All derivatives were solubilized in 100% DMSO and administered at a final concentration of 143 μM per well for 45 minutes, washed 3 times with media. 45-minute exposure mimics the exposure time in a human. Each well contained 10 male adult worms. All screens were performed in experimental and biological triplicate. Survival was plotted as a percentage over time using Prism/Curve Comparison/ Long-rank (Mantel-cox) test. The p-value threshold for each derivative compared to DMSO was <0.001.
Fig 6.
OXA Derivatives Significantly Reduced the Number of Collected S. mansoni Juvenile Worms from Infected Mice.
Five mice per group were infected with 100 cercaria of S. mansoni. Mice were treated with a single dose of 100 mg/kg by oral gavage of CIDD-0150303 at the day specified on the X-axis and worm burden determined on day 45 pi. Prism/unpaired t test (P<0.05).
Fig 7.
Crystal Structures of CIDD-0150303 (Left Panel) And CIDD-0150610 (Right Panel) Shown in The Active Site of SmSULT.
The active site inner surface cavity is depicted in light green. Secondary structure elements in front of the compounds were omitted for clarity. The branched indole and (trifluoromethyl) phenyl groups at top are observed in swapped positions between the two compounds.
Fig 8.
Kaplan-Meier Curves Demonstrate the Ability of CIDD-0149830, CIDD-0150610, and CIDD-0150303 to Kill S. mansoni PZQ-R.
All derivatives were solubilized in 100% DMSO and administered at a final concentration of 143 μM per well for 45 minutes, washed 3 times with media. 45-minute exposure mimics the exposure time in a human Each well contained 10 male adult worms. All screens were performed in experimental and biological triplicate. Survival was plotted as a percentage over time using Prism/Curve Comparison/ Long-rank (Mantel-cox) test. The p-value threshold for each derivative compared to DMSO was <0.001.
Fig 9.
Combination Therapy of PZQ-R Infected Mice.
Five mice per group were infected with S. mansoni PZQ-R. Worms were collected 10 after days of treatment with a single dose of 100 μg/g by oral gavage and compared to the control group. Prism/unpaired t test (P<0.05).