Supporting Information
Bi-Functional Elastin-Like Polypeptide Nanoparticles Bind Rapamycin and Integrins, and Suppress Tumor Growth In Vivo. **Jugal P. Dhandhukia1, **Pu Shi1, Santosh Peddi1, Zhe Li1, Suhaas Aluri1, Yaping Ju1, Dab Brill1, Wan Wang1, Siti Janib1, Yi-An Lin2, Shuanglong Liu3, Honggang Cui2, J. Andrew MacKay1,4,5* 1Department
of Pharmacology and Pharmaceutical Sciences, University of Southern
California School of Pharmacy, Los Angeles, CA 90089, USA 2Department
of Chemical and Biomolecular Engineering, Johns Hopkins University,
Baltimore, MD 21218, USA 3Department
of Radiology, Molecular Imaging Center, University of Southern California
Keck School of Medicine, Los Angeles, CA 90033, USA 4Department
of Biomedical Engineering, University of Southern California Viterbi School of
Engineering, Los Angeles, CA 90089, USA 5Department
of Ophthalmology, University of Southern California Keck School of Medicine,
Los Angeles, CA 90033, USA **Author’s contributed equally towards work. *Corresponding author: J. Andrew MacKay (
[email protected]) Telephone: 1-323-442-4118 Address: 1985 Zonal Avenue, PSC 306A, School of Pharmacy, University of Southern California, Los Angeles, CA 90089, USA
Figure S1: Co-assembly of ISR and FSI diblock copolymers in different ratios does not affect the CMT or particle size. The optical density for mixtures of 10 µM ISR + 190 µM FSI, 20 µM ISR + 180 µM FSI, 50 µM ISR + 150 µM FSI, 100 µM ISR + 100 µM FSI and 200 µM FSI were measured using both UV-Vis spectrophotometry and Dynamic Light Scattering over a temperature gradient. Bifunctional nanoparticles mixed at varying ISR concentrations exhibit (a) similar CMT values and (b) similar hydrodynamic radii.
Figure S2: Body weight distribution in tumor regression studies after start of treatment. No change in body weight was observed during the entire course of treatment in (a) tumor regression study with free Rapa, SI-Rapa at 0.25 mg/kg dose, and dose escalation of FSI-Rapa from 0.0075-0.25 mg/kg and (b) comparative tumor regression study with free Rapa, FSI-Rapa and ISR/FSI-Rapa at 0.075 mg/kg dose; which indicates the treatments were well tolerated and no significant cytotoxicity was associated with any of the formulations.
Figure S3: microPET imaging demonstrates higher whole body distribution for ISR/FSI over FSI. 64 Cu-AmBaSar labeled proteins were administered intravenously to orthotopic MDA-MB-468 breast tumor implanted nude mice (n = 8-11). Serial imaging was performed and images for FSI and ISR/FSI are shown from 5 min to 8 h. A representative mouse is shown from each group. Heart and tumor accumulation is shown by yellow and white arrows respectively.
Figure S4: FSI protein with or without ISR gives a similar bio-distribution and pharmacokinetic halflife. 64Cu-AmBaSar labeled proteins were administered intravenously to orthotopic MDA-MB-468 breast tumor implanted nude mice (n = 8-11). Serial images (Figure S3) were quantified within (a) heart, (b) kidneys, (c) liver, (d) tumor, (e) lungs, (f) muscle and (g) spleen expressed as % ID/g calculated from ROI images analysis. Values indicate mean ± SD (n = 8-11 mice).
Figure S5: Direct covalent labeling with rhodamine destabilizes FSI nanoparticles. FSI was covalently labeled with rhodamine and stability was evaluated by measuring Rh at 37 °C for a period of 12 h. The Rh of FSI-Rho was found to increase by 4 h post labeling indicating instability. The backbone ELP, SI was labeled at its single amino terminus and co-assembled with FSI or FSI/ISR. The co-assembled nanoparticles were stable for a period of at least 12 h post labeling.
