PMC

HPLC chromatogram of ¹²⁵I-ANG1005 in perfusion fluid (A) and brain (B) after perfusion uptake for 120 s. Intact ¹²⁵I-ANG1005 eluted at 13–15 min. Tracer integrity in brain of tracer was >95%.

Effect of capillary depletion (A), post perfusion wash for 1 min with neutral saline (pH 7.4), cold acidic saline (pH 3), or 2.7% human serum albumin fluid (B), or neutral saline wash for 1–15 min (C) on brain ¹²⁵I-angiopep-2 (A) or ¹²⁵I-ANG1005 (A,B, and C) after 120 s of uptake perfusion. Data represent mean ± SEM (n= 3–5 per group). * P < 0.05

Time course (A) of brain uptake of ¹²⁵I-ANG1005, ¹²⁵I-angiopep-2, and ³H-paclitaxel during in situ brain perfusion. Data represent mean ± SD (n = 3–6 per group). Lines represent best fits to the data by least squares regression. Regional brain uptake (B) of ¹²⁵I-ANG1005 during in situ brain perfusion with tracer saline. Data represent mean ± SEM (n=5). Statistical significance was determined using ANOVA (P>0.05) for all seven well perfused regions.

Distribution of green fluorescent protein transfected 231-BR metastases of breast cancer (Green: A and C), Texas Red 3kD dextran (Red: A and C), ¹²⁵I-ANG1005 (Quantitative autoradiograph, B), and ¹⁴C-paclitaxel (Quantitative auroradiograph, D) in 20 μm coronal representative sections of brain. ¹²⁵I-ANG1005 and ¹⁴C-paclitaxel were allowed to circulate in vivo after i.v. injection for 30 min. Texas Red dextran circulated for 10 min prior to death. Animals received equivalent drug doses; radioactivity was normalized to 7.5 μCi/mouse.

Brain and brain metastasis molar (nmol/kg) concentrations of paclitaxel and ANG1005 at 30 min after injection in vivo in mice. Results were normalized to an equivalent drug dose of 14 μmole/kg mouse (i.e., the dose for paclitaxel) and are expressed in nmole/kg wet tissue weight. A – Represents total tissue concentrations of paclitaxel or ANG1005; B – represents vascularly corrected tissue concentrations of paclitaxel or ANG1005; and C – represents vascularly corrected tissue concentrations expressed in terms of paclitaxel content (note - each mole of ANG1005 carries 3 moles of paclitaxel). Results are expressed as mean ± SEM (n= 11–25). * P < 0.05 brain vs tumor for ANG1005. *** P < 0.05 paclitaxel vs ANG1005 within the same tissue.

A - Effect of co-perfusion with unlabeled angiopep-2, LRP ligands – aprotinin (10 μM) or RAP (200 nM), or cold temperature (4 °C) on the brain uptake of ¹²⁵I-ANG1005. B - Effect of plasma protein (2.7% albumin) on brain uptake of ¹²⁵I-ANG1005. C – Effect of transport inhibitors or competitors, including L-phenylalanine (large neutral amino acid transport), poly-L-lysine (cationic absorption), and chlorpromazine and indomethacin (caveolae- and clathrin-mediated endocytosis), on BBB transport of ¹²⁵I-ANG1005. Data represent mean ± SEM (n= 3–5 per group). Statistical significance was determined using ANOVA with Dunnett’s post hoc test in A and Student’s t-test in B. * P < 0.05. ** P < 0.01.