Biodistribution, pharmacokinetics and toxicity of a Vasconcellea cundinamarcensis proteinase fraction with pharmacological activity

P1G10, the Proteolytic Fraction from Vasconcellea cundinamarcensis, Stimulates Tissue Repair after Acute Exposure to Ultraviolet B Radiation

“…The first protein fraction (P1G10) containing the bulk proteolytic activity was pooled and concentrated by ultrafiltration (10,000 Da pore size) and stored at −20 °C until used. The uniform composition of P1G10 has been demonstrated earlier by HPLC and SDS-PAGE electrophoresis [46] and the peptide isoforms comprising P1G10 were described [47]. The standard protein concentration and amidase activity of this fraction were 8.39 ± 0.39 mg/mL and 13.5 ± 0.5 nM/μg·×·min, respectively.…”

“…The protein was extracted in 500 µL ice-cold lysis buffer (20 mM Tris–HC1, pH 7.5, 150 mM NaCl, 1 mM EDTA, 1 mM EGTA, 1% Triton X-100, 2.5 mM sodium pyrophosphate, 1 mM b-glycerophosphate, 1 mM Na 3 VO 4 , 1 mg/L leupeptin, 1 mM PMSF—Sigma-Aldrich). After centrifugation at 10,000× g at 4 °C for 30 min, the protein concentration was assessed by Bradford assay [46,51]. The proteins (30 µg) were resolved on a 10% SDS–PAGE and electroblotted onto PVDF membranes.…”

Cysteine Proteases from V. cundinamarcensis (C. candamarcensis) Inhibit Melanoma Metastasis and Modulate Expression of Proteins Related to Proliferation, Migration and Differentiation

“…We showed that a cysteine proteinase enriched fraction from V. cundinamarcensis , named as P1G10, displays a variety of pharmacological activities, such as fibrinogenolytic, fibrinolytic and antithrombotic activities [ 2 ], and tissue repairer effect in cutaneous [ 3 , 4 , 5 ], and gastric lesions [ 6 , 7 ]. Preclinical studies show that P1G10 is non-toxic when used topically [ 4 ] or systemically, with no evidence of mutagenic or genotoxic effects [ 8 ]. We recently reported that P1G10 displays a remarkable antitumor activity in a murine melanoma model.…”

Antiangiogenesis, Loss of Cell Adhesion and Apoptosis Are Involved in the Antitumoral Activity of Proteases from V. cundinamarcensis (C. candamarcensis) in Murine Melanoma B16F1

“…In most situations the biological activity of plant cysteine proteinases is explained by the hydrolytic activity, immunomodulation, and/or binding to circulating anti-proteases, upsetting the homeostasis of endogenous circulating proteinases [ 10 ]. Furthermore, the proteolytic mixture lacks significant dermal and systemic toxicity [ 6 ] and no mutagenic or genotoxic effects are evident [ 11 ]. In support of the enhanced wound healing activity, Gomes and coworkers [ 12 , 13 ] demonstrated that two of the proteinases in P1G10, (CMS2MS2 and CMS2MS3), display mitogenic activity on fibroblasts and other cell lineages, an effect mediated by the activation of MAP kinase pathway and which is independent of the proteolytic activity.…”