Fig 1.
Molecular structures of MCL, DMAMCL, and DMAMCL salts.
Fig 2.
Biodistribution of DMAMCL administered orally.
DMAMCL concentrations over time in rats after a single oral dose (100 mg/kg) (n = 6) in plasma, brain, spleen, lung, kidney, heart and liver were detected by UPLC-MS/MS. Statistical significance between plasma and brain is indicated by asterisks (*p < 0.01 compared to vehicle; **p < 0.01 compared to vehicle).
Table 1.
Mean concentration of DMAMCL in plasma and brain at different time after administration.
Fig 3.
Inhibition of glioma cell growth by DMAMCL.
C6 and U-87MG cells were treated with different concentrations of DMAMCL for 72 h. Cell viability was measured by MTT assays.
Fig 4.
C6 and U-87MG cells were treated with DMAMCL. After 24 h, cells were collected for Annexin V and PI detection or total protein extraction. (A) Representative FACS results. (B) and (C) The percentage of cell apoptosis (the total of early and late apoptosis) for C6 and U-87MG cells, respectively. (D) and (E) Western blotting detection for Bax, Bcl-2 and β-actin protein expression for C6 and U-87MG cells, respectively. One-way ANOVAs followed by Tukey’s post-hoc tests were used and statistical significance is indicated by asterisks (*p < 0.01 compared to vehicle; **p < 0.01 compared to vehicle; ***p < 0.001 compared to vehicle).
Fig 5.
Treatment of intracerebral C6 glioma with DMAMCL.
After euthanasia, brains and tumors were photographed, and tumors were weighed. Photographs of (A) whole brains and (B) tumors are shown. (D) Changes in rat body weight over time and (C) distributions of tumor weight for each group at the end of the experiment. Average tumor weight for each group is indicated by red lines. Statistical significance is indicated by asterisks (**p < 0.01 compared to vehicle; ***p < 0.001 compared to vehicle). E) Kaplan-Meier survival curves over 63 days. Rats were treated daily for 21 days with vehicle, VCR, or DMAMCL at 25, 50, or 100 mg/kg. (n = 10 rats/group). All groups were compared with the vehicle group. Significance is indicated by asterisks (**p < 0.01, ***p < 0.001).
Table 2.
DMAMCL inhibition of intracranial C6 tumor growth.
Table 3.
Effects of DMAMCL on C6 tumor-bearing rat survival.
Fig 6.
HE staining of brains from tumor-bearing rats after treatment for 21 days.
All images are representative images, and scale bars indicate 100 μm.
Fig 7.
Toxicology evaluation of DMAMCL.
A) and B) Effect of DMAMCL on body weight and food intake. DMAMCL (200 or 300 mg/kg) was orally administered once a day for 21 days and body weights were measured on days 0, 3, 7, 11, 14, 17, and 21 (n = 6 rats per group). Total food intake was measured as the weight of food consumed during a 24-h period on days 3, 8, 15, and 22. C) Locomotor activities of rats Rats were orally administered with or without DMAMCL (either 100 or 300 mg/kg) once a day for 21 days. The individual total distance of rats was monitored during a test period of 60 min on days 1, 10, 21 and 28. Statistical significance is indicated by asterisks (*p < 0.05 compared to vehicle). D) HE staining of rat tissues after daily treatment with vehicle or DMAMCL at 300 mg/kg for 21 days. Representative HE staining results are shown. Scale bars indicate 100μm.
Table 4.
Effects of DMAMCL on hematological parameters.
Table 5.
Effects of DMAMCL on the serum biochemistry of rats.