Pharmaceutical angiostatic dipeptide compositions and methods of use thereof
a technology of angiostatic dipeptide and composition, which is applied in the direction of peptide/protein ingredients, drug compositions, angiogenin, etc., can solve problems such as the problematic clinical experience of cytokine therapy, and achieve the effects of stimulating the production of immune cells, normalizing numerical relationships, and low levels of l-glu-l-trp dipeptide preventing neovascularization
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example 1
Lack of Mutagenicity and Toxicity of L-Glu-L-Trp: Pharmacokinetics and Biodistribution
[0058] *Note that general materials and methods used in Examples, below, appear at the end of the Examples section and immediately before the citations.
Protocol A
Acute Toxicity Studies
[0059] Summary: L-Glu-L-Trp when injected im at dosages calculated to be about 10,000-times a therapeutic dosage were non toxic in mice, guinea pigs, chickens, and dogs as determined by monitoring general condition, behavior, movements, cardiac and respiratory physiology, and gross pathology.
Protocol B
Chronic Toxicity Studies
[0060] Summary: L-Glu-L-Trp when injected daily as a single im or iv for a period of 28 days was without adverse effects as determined by monitoring behavior, feeding, body weight, coat condition, mucous membranes, red and white cell blood counts, cardiac and respiratory physiology, liver and kidney function, and gross pathology. Kidney function was determined by evaluation of diuresis afte...
example 2
Inhibitory Effects of L-Glu-L-Trp on Angiogenesis in Chorioallantoic Membrane (CAM) Assays
[0062] Briefly, eight-day chicken embryos were removed from eggs and placed into sterile petri dishes. Individual filter paper disks were saturated with 7.5 μl of different stock solutions of L-Glu-L-Trp dissolved in sterile 0.14M NaCl to achieve final test concentrations of 0.001, 0.01, 0.1, 1.0, 10, 100, 500, and 1000 μg per disk. Disks were air dried and then inverted onto the surface of the respective embryos. Embryo vascularization was assessed after 48 hrs. of incubation using the grading scale summarized in TABLE 1, below.
TABLE 1Scoring of Chicken Embryo Vascularity: CAM AssayPercentageInhibition GradeDescriptionInhibition0Not visibly difference than negative0control1Slight inhibition of vessel formation252Moderate inhibition of vessel formation503Near-complete inhibition of vessel75formation4Complete inhibition of vessel formation100
[0063] In this experiment saline served as a negati...
example 3
Inhibition of Neovascularization of Lewis Carcinoma
[0065] Lewis lung carcinoma cells (5×107) when injected (0.1 ml) intradermally into both flanks of C57BL / 6 mice (day 0) produce a visible highly vascularized tumor nodules within 7 days. By excising the tumor the degree of tumor vascularity may be determined microscopically by counting the number of large vessels radiating from the tumor mass. An independent study was performed (as follows) at a GLP approved contract research organization.
[0066] Saline was used as a negative control and Cytoxan as a positive control. The positive control, Cytoxan (200 mg / kg), was administered only on day 2. Test treatments with L-Glu-L-Trp were administered im on a daily basis starting on day 1 after tumor injection and continuing for 5 days (i.e., through day 6). L-Glu-L-Trp was administered at doses of 125, 250, 500, 1000, and 2000 μg / kg / dose. The negative control, saline, was administered ip on the same daily 5 day schedule. Ten mice (20 tumors...
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