Read carefully (i read synergy and look at those claims)
>EXAMPLE I-Treatment with an LFA-I antagonist and a TNF antagonist
DBA-IJ mice were immunized with 100 ug bovine collagen type 11 in 100 ul complete Freund's adjuvant (CFA) followed by a second injection of the same collagen in incomplete Freund's adjuvant (IFA) 21 days later. The collagen type 11 in CFA or IFA was injected intradermally at the base of the tail.
Animals were evaluated every other day. At the onset of arthritis in any of the animals, as noted by swelling of any of the paws. treatment was initiated in all animals placed randomly into the following treatment groups with 12 mice per group.
Group 1. Control: Treatment with saline. 100 ul, intraperitoneal route. every day for 14 days. then 3 times per week every other day (Monday. Wednesday and Friday).
Group 2. Anti-murine CD la monoclonal antibody M17. 150 ug (approx 8 mg/kg) via intraperitoneal route given at onset of disease followed by 3 times per week every other day (Monday. Wednesday and Friday) until the end of the study.
Group 3. ENBREL (human TNF-Fc. 50 ug) intraperitoneal route, given daily at onset of disease for 14 days.
Group 4. Combination of ENBREL and Anti-CD la : Anti-murine CD ! la mab M17, 150 ug (approximately 8 mg/kg) via intraperitoneal route given at onset of disease followed by 3 times per week every other day (Monday. Wednesday and Friday) until the end of the experiment. ENBREL, intraperitoneal route, given daily at onset of disease for 14 days.
After the onset of treatment, the mice were evaluated every other day 3 times a week and the severity of paw swelling was subjectively graded for each paw on a scale of 0-4 with 0 = normal, I = minimal, 2 = mild, 3 = moderate, and 4 = severe. A cumulative score was recorded for each animal (potential range 0-16). The animals were terminated 38 days after the initiation of treatment. Radiographs were take on of all four limbs to evaluate for joint lesions and the paws were collected for histopathology. The severity of disease as determined bs clinical score for each group (mean +/-standard deviation) was graphed and compared between groups. The clinical scores taken on the last day were corroborated by histological and radiologic analysis of all four paws done at the terminus of the study.
The use of either ENBREL or anti-CD11 a antibody reduced the clinical scores compared to the control group (p < 0.05) and the combination of anti-CDI la antibody and ENBREL improved the clinical scores compared to ENBREL alone (p < 0.05). See Figure I.
EXAMPLE 2-Treatment of arthritis with an LFA-I antagonist or a TNF antagonist
In this example. arthritis was induced in DBA-lLacJ mice (Figure 2) or DBA-IJ mice (Figure 3) which were then treated with anti-murine CDI la antibody (M17), ENBREL, or saline as a control. These experiments were performed as described in Example I and in the inset in Figure 2 and Figure 3. Treatment was initiated on day 48 or day 22 post immunization, the day of onset of arthritis in the experiments of Figure 2 and Figure 3. respectively.
The results presented in Figure 2 and Figure 3 show that anti-CD la antibody or ENBREL alone is effective in treating arthritis as evidenced by the reduction in clinical scores.
EXAMPLE 3-Treatment of arthritis with an LFA-1 antagonist and a TNF antagonist
In this example. arthritis was induced in DBA-ILacJ mice (Figure 4) or DBA-IJ mice (Figure 5) which were then treated with anti-murine CD I 1 a antibody (M 17) alone. ENBREL alone. saline as a control. or a combination of M 17 and ENBREL. These experiments were performed as described in Example I and in the inset in Figures 4 and 5. In Figure 4. treatment was initiated on dav 40 post immunization. M 17 was given at
160 ug. three times per week for the duration of the study. For the combination therapy. the mice received 50 us
Enbrel daily up to a total of 14 doses in one experiment, and for the duration of the study in another experiment.
In Figure 5. treatment was initiated on day 24 post immunization. the day of onset of arthritis. Enbrel was administered everyday (qd) for 14 days. then every other day (qod). (Monday, Wednesday, Friday) until the end of the experiment.
As is evident from the results shown in Figure 4 and Figure 5, combination therapy with an LFA-I antagonist and a TNF antagonist had a svnergistic effect over treatment with either antagonist alone. resulting in greater reduction in mean clinical scores to almost normal in this animal model.
In Examples 4-6 below. a test compound refers to an LFA-1 antagonist (e. g.. anti-CD la antibody) or a
TNF antagonist (e. g., ENBREL). The volumes, concentrations and time points are exemplary and can be varied as will be familiar to one of skill in the art. |
