"FORMULATED TEST MATERIAL STABILITY: From a single preparation, samples for stability analysis were collected immediately after the first preparation and 7, 14, 21, 28, and 32 days from the date of the first preparation and frozen at -20°C. Data for the stability of the formulated test material is included in Appendix 9. With the exception of Study Day 1, stability data indicate the actual versus the theoretical concentrations for 4, 12, 40 mg/ml were within 10%. On Study Day 1, the 4 and 12 mg/ml actual values were 14% and 32% of their theoretical concentrations, respectively. It is unclear why these values are outside of 10% theoretical concentrations but these data support that at 540 mg/ml T-7485 does not decline but is stable for 32 days after the initial formulation.

MORTALITY: The summary of mortality is in Table 1. The individual observations are in Appendix 3. Observations for mortality and moribundity were recorded twice daily (a.m. and p.m.). During the study, there were no mortality recorded at any level of treatment for either male or female rats.

CLINICAL OBSERVATIONS: The summary of clinical observations is in Table 1. The individual observations are in Appendix 3. Clinical observations were recorded once daily, and no adverse clinical observations were recorded at any level of treatment for either male or female rats.

BODY WEIGHTS AND BODY WEIGHT CHANGES: The summaries of body weights and body weight changes are in Tables 2 and 3, respectively.· The individual body weights are in Appendix 4. Body weights were recorded on Study Days 1, 7, 10, and 11 . There were no significant effects of T-7485 treatment on male or female body weights. On Study Day 10, mean male body weights ranged between 275.8 and 286.2 grams, while female rats ranged between 193.2 and 206.2 grams. There were no significant effects of T-7485 treatment on mate or female rat body weight changes. Between Study Days 1 and 10, the range of mean male body weight change was 62.4 to 75.0 grams, while in females the range was 27.8 to 32.4 grams.

FEED CONSUMPTION: The summary of absolute feed consumption is in Table 4. The individual data are in Appendix 5. Feed consumption was measured on Study Days 1, 7, and 10. Relative to body weights and body weight changes, there was no adverse effect of T-7 485 on absolute feed consumption for either male or female rats. On Study Days 1 through 10, the mean feed consumption ranged in males from 28.0 to 29.5 grams/day, while in female rats the mean feed consumption ranged from 20.8 to 12.9 grams/day.

HEMATOLOGY: The summary of hematology data is in Table 5. The individual hematology data are in Appendix 6. Values in parenthesis indicate mean ± one standard deviation. Blood samples for hematological analysis were obtained via cardiac puncture prior to necropsy. The animals were anesthetized with carbon dioxide for the collection. Feed was withheld overnight prior to collection. All hematological measurements in test material-treated groups were comparable to the vehicle control with the exception of elevated segmented neutrophils (1.2 ± 0.34103/mm3 vs. 0.5 ± 0.25 103/mm3 vehicle control) in females that received 1000 mg/kg/day T-7485. This elevation in segmented neutrophils appears to be an isolated change that occurred only in one gender and had no clear association with other reported findings (including histopathology). It is therefore, unclear if this marginal change is related to the test material.

CLINICAL CHEMISTRY: The summary of clinical chemistry data is in Table 6. The individual clinical chemistry data are in Appendix 7. Values in parenthesis indicate mean± one standard deviation.

• At 100 mg/kg/day/day of T-7485, male mean albumin levels were significantly decreased when compared to the vehicle control (3.9 ± 0.39 vs. 4.4 ± 0.10 g/dl, respectively).

• At 300 mg/kg/day of T-7485, male mean gamma-glutamyltransferase (GGT) was significantly elevated when compared to the vehicle control {4 ± 2.4 vs. 0 ± 0 U/L, respectively).

• At 100 and 300 mg/kg/day T-7485, male mean chloride levels were significantly elevated when compared to the vehicle control (89 ± 2.2, 90 ± 1.5, and 87 ± 1.1; respectively).

• At 300 mg/kg/day T-7485, female mean alkaline phosphatase levels were significantly decreased when compared to the vehicle control (140 ± 41.2 vs. 221 ± 26.3, respectively).

• At 1000 mg/kg/day T-7485, female mean glucose levels were significantly increased when compared to the vehicle control (163 ± 23.5 vs. 107 ± 17.1 mg/dl, respectively). In summary, with the exception of the elevated glucose level in females that received 1000 mg/kg/day, T-7485 had no dose-dependent effect on clinical chemistry parameters. The elevated glucose level was an isolated, gender specific change and it is unclear if this is related to the test material.

NECROPSY: The individual necropsy observations are included in the pathology report in Appendix 2. Necropsy observations included: diverticulum of the small intestine, liver foci, small seminal vesicles, enlarged mandibular lymph nodes, lung foci, and kidney dilatation. Many of the gross findings correlated with microscopic findings. Three singular incidences of enlarged mandibular lymph nodes were noted in males that received vehicle control and females that received 300 or 1000 mg/kg/day T-7485. Enlarged mandibular lymph nodes correlated with either mild lymphoid hyperplasia (vehicle control male and high dose female, respectively) or mild plasmacytosis (females that received 300 mg/kg/day T-7485). In males that received 100 mg/kg/day T-7485, kidney dilatation and diverticulum of the small intestine were correlated microscopically with unilateral moderate hydronephrosis and diverticulum, respectively. In males that received 1000 mg/kg/day T-7485, lung foci was correlated microscopically with moderate focal hemorrhage. Finally, in females that received 1000 mg/kg/day T-7485, lung foci were correlated microscopically with mild chronic focal inflammation accompanied by mild focal crystalline material.

ORGAN WEIGHTS: The summary of organ weights is in Table 7. The individual organ weights are in Appendix 8. Organ weights were recorded for the following organs prior to fixation: adrenal glands, brain,

heart, kidneys, liver, ovaries, spleen, testes, and thymus. Paired organs were weighed together. Absolute and relative organ weights were similar to the corresponding vehicle control groups for both male and female rats with the exception of absolute and relative liver weights. Mean percent increases in absolute, % body weight, and % brain weight liver weights in males (female percentages are in parenthesis) that received 1000 mg/kg/day/day were 36% (22%), 34% {18%), and 40% {22%), respectively.

HISTOPATHOLOGY: The pathology report is in Appendix 2. Histopathology was performed on all tissues from the vehicle control and high-dose group animals and gross lesions from all animals. Non-neoplastic microscopic lesions included the following: necrosis of the heart; inflammation and hemorrhage of the esophagus; inflammation and edema of the stomach; ultimobranchial cyst of the thyroid; lymphoid hyperplasia and plasmacytosis of the mandibular lymph nodes; hemorrhage of the thymus; inflammation, hemorrhage, and crystalline material of the lung; and tubular degeneration, mineralization, and hydronephrosis of the kidneys. None of the microscopic lesions appear to be related to administration of the test material. The most common lesion was focal/multifocal necrosis of the cardiac muscle of the heart. This necrosis occurred in both the vehicle control rats and rats that received 1000 mg/kg/day T-7 485. This lesion probably represents early cardiomyopathy, which is a common spontaneous lesion in Sprague-Dawley rats."

Mononuclear Cell Infiltrate

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