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Exam le A phase 2 trial was conducted at San Francisco General Hospital. The study enrolled 18 patients in an open label pathogenesis study of WF-10. Patients received one hour infusions of WF-10 for one week, followed by two weeks of rest. On the third week, the patients again received one hour infusions of WF-10 daily for one week followed by two weeks of rest. Parameters studied included mesures of macrophage activation/function immunologic activation and HIV viral load. RBC hemolysis evaluation studies included 51 Cr-RBC survival studies compare with changes in hemoglobin, haptoglobin and reticulocyte values.
There were no side effects noted in any of the 18 patients. Data on eight of the patients were gathered and the results are tabulated below, and depicted in Figures 6-13. There appeared to be acute increases in the following parameters as measured by flow cytometry (FACSCAN as recommended by, for example, Becton-Dickinson) in relation to drug administration, changes that generally returned close to baseline within 2 weeks of drug administration : CD-4, CD-8, CD14+/CD69+, CD14+ side scatter, CD20/DR+ cells. Several values seemed to generally increase through the study, showing no clear downward trend by the end of the study and may represent long-term changes induced by WF-10. These include an increase in macrophage phagocytosis index and an increase in the CD3+/CD8+/CD28-subset of T-cells.
Potential downward trends were noted in the following categories : macrophage intracellular TNF-α secretion, and a decrease in the number of circulating CD14+/DR+ cells. It has been reported that immune paralysis results when the number of circulating CD14+/DR+ cells decreases to such an extent as to reach a threshold value. No obvious changes were noted in T-cell PHA activation values or HIV load as measured by the HIV bDNA assay (most of the patients had no detectable HIV thoughout the study). Results of the RBC survival studies showed no evidence for hemolysis in response to the treatment.
As shown in Figure 6, administration of WF10 results in an increase in CD14+/CD69+ cells, with dramatic increases immediately following infusion.
Figure 7 shows a decrease in CD14+/TNF secretion after administration of WF10, thereby indicating that a stabilized chlorite solution is effective in decreasing secretion of the tumor necrosis factor cytokine.
Figures 8 and 9 show that administration of WF10 to patients in vivo results in a steady increase in the number of CD3 +/CD8+, as well as a steady increase in the number of CD3+/CD8+/CD28-T cells. The in vitro data above show inhibition of antigen presentation using CD4+ T cells, and Figures 8 and 9 show an increase in the number of circulating CD28-T cells (CD3+ T cells). Figure 10 illustrates an increase in phagocytosis index upon administration of WF10. Figure 11 shows a decrease in immune function upon administration of WF10 by virtue of the decrease in CD14+/DR+ cells. The inventors therefore believe that the stabilized chlorite solution of the invention is capable of up-regulating phagocytosis, while at the same time, down-regulating or suppressing the cell-mediated and humoral immune response.
The results tabulated below summarize the data from 15 patients and show the changes in various measured parameters between the 8th day and the 47 day of treatment. The 8 day represents the first day of WF10 administration because the first 7 days of treatment are devoted to patient evaluation. Parameter Measured p-value* Direction CD3+, CD8+, CD28-0. 027 increase CD14+, TNF-0. 017 decrease CD14+, DR+ 0. 032 decrease CD3+, CD4+, CD38+ (MF CD38 Antigen) 0. 021 decrease CD3+, CD8+, CD28+ (MFCD28 Antigen) 0. 010 decrease Cl20, DR+ (MF DR Antigen) 0. 014 decrease All CD14+ 0. 037 decrease -One-tailed p-value. Sample size of 15 patients using Wilcoxon rank statistic.
These data show that administration of WF10 in vivo to humans shows an increase in the production of CD28-subset of CD8+ T-cells. The data also show an increase in macrophage activation leading to phagocytosis. The data further show no evidence of RBC hemolysis. When coupled with the in vitro studies showing the inhibition of antigen presentation for CD4+ cells, it is believed that administration of WF-10 in vivo will result in inhibition and/or prevention of antigen presentation in APC, as well as stimulate macrophage activation resulting in increased phagocytosis.
Examle 6 Based on the in vivo data above, administration of WF10 has shown a consistent down regulation of CD14+/DR+ cells achieving statistical significance.
In addition, WF10 administration in vivo has shown overall reduction of CD3+/CD8+/CD28+ cells, and significant increased levels of CD3+/CD8+/CD28- cells of long-ter duration. The in vitro data above also show that WF10 is effective in inhibiting and/or preventing antigen presentation. This reduced antigen presentation may be critical in inhibiting lymphoproliferative disease, and in particular in inhibiting B-cell lymphoma and thus, it is expected that WF10 therapy will be effective for treatment of lymphoma. In accordance with this expectation, in the case of a single patient suffering from B-cell lymphoma, the patient responded to WF10 therapy with a notable reduction of tumor size with no recurrence to date.
Adult patients having low grade follicular lymphoma are selected based on their lack of cnrollment in current therapy regimens. Fifteen patients having lymph nodes > 1 cm
in diameter at baseline confirme by CT scan will be enrolled in an open-label, single arm, single center study. Patients will receive periodic 0. 5 ml/kg infusions of WF10 from days 1-5 (week 1) and days 8-12 (week 2). After screening evaluations are completed (about 14 days), eligible patients will attend pre-study visit in week 0 to acquire the baseline data.
Screening criteria include the following : male or female patients greater than 18 years of age ; histologically confirme follicular lymphoma ; mesurable disease defined as having lymph nodes > 1 cm
in diameter as measured by CT ; adequate renal function documente by a serum creatinine < 2 times in institution's ULN ; adequate liver function documente by a serum billrubin less than or equal to 1. 5 mg/dl and SGOT (AST) or SGPT (ALT) < 5 times the institutional upper limit of normal ; written informed consent to participate in this study and a willingness to comply with all procedures and scheduled visits ; hemoglobin > 9. 0 g/dl for woman and > 10. 0 g/dl for men ; platelet count > 75, 000/mm2 ; and absolut neutrophil count > 750/mm2.
WF10 will be applied at a dose of 0. 5 ml per kg of body weight diluted into 250 to 500 ml normal saline administered by intravenous infusion of 1 hour duration. CT measurements will be taken to determine tumor size at week 0, on day 15, day 30 and day 45. Follow-up period will last for a duration of 3 months with final CT measurements on day 90.
CT measurements reveal that administration of WF10 results in a reduction of lymph node size. Patients also exhibit an increase in CD3 +/CD8+/CD28, an increase in CD14+/DR+ and an increase in CD40 T cell subsets.