Extended Data Fig. 7: In vitro characterization of SHA-E2 and in vivo release of ganfeborole.
a) Chemical structure of SHA-E2 with the mass spectrum and UPLC chromatogram of the material confirming the identity and purity of the peptide. b) Mass spectrum and UPLC chromatogram of the unmodified E2 peptide. c) Frequency sweep collected by oscillatory rheology shows that SHA-E2 hydrogels are more frequency dependent than unmodified E2 and form slightly weaker gels, as indicated by a reduction in the difference between the storage modulus (G′), indicated by filled circles and solid lines, and the loss modulus (G″), indicated by open circles and dotted lines. d) Representative histological sections of SHA-K2 and SHA-E2 gels excised three days after subcutaneous injection. Sections were stained with hematoxylin & eosin (left two images) or Masson’s trichrome (right four images) stains. Black 750 μm scale bar applies to the left four full sized images. Histological analysis shows that SHA-K2 gels swell significantly and are heavily infiltrated by cells, consistent with an inflammatory response. SHA-E2 remains minimally infiltrated by cells, has less collagen deposition on the border of the gel, and does not swell in size. e) Cumulative release of ganfeborole after 24 h from SHA-K2 and SHA-E2 are statistically similar, suggesting that changing the peptide used in SABER hydrogels does not compromise its ability to control the release of BA-containing small molecules. Data is presented as the mean of three replicates ± SD. f) Pharmacokinetic parameters extracted by performing a non-compartmental analysis on the in vivo release of ganfeborole from SHA-E2 show that using the SABER hydrogel improves drug exposure (AUC), half-life (t1/2) and reduces the maximum circulating concentration (Cmax) of the compound. Pharmacokinetic parameters are presented as the mean of n=4-5 replicates ± SD.
