There are quantitative and/or qualitative mechanisms allowing
androgen receptor (AR) growth signaling in
androgen ablation refractory
prostate cancer cells. Regardless of the mechanism, agents that deplete AR
protein expression prevent such AR growth signaling.
Thapsigargin (TG) is a highly cell-penetrant sequiterpene-
lactone that once inside cells inhibits (IC(50), ∼ 10 nmol/L) critically important housekeeping SERCA 2b
calcium pumps in the endoplasmic reticulum. Using a series of five genetically diverse
androgen ablation refractory human
prostate cancer lines (LNCaP, LAPC-4, VCaP, MDA-PCa-2b, and CWR22Rv1), TG inhibition of SERCA pumps consistently results in depletion of the endoplasmic reticulum Ca(+2) coupled with μmol/L elevation in the intracellular free Ca(+2) initiating a molecular cascade that: (a) inhibits Cap-dependent AR
protein synthesis resulting in 90% depletion of AR
protein by 24 hours of TG exposure, (b) arrests the cells in G(0), and (c) induces their apoptotic death. Unfortunately, due to its highly lipophilic nature, TG is not deliverable as a systemic agent without host toxicity. Therefore, TG analogues containing
amino acids were developed, which retain ability to deplete AR
protein and induce cell death and which can be covalently linked to
peptide carriers producing water soluble
prodrugs for systemic delivery. Specific amino acid sequences are used to restrict the liberation of cytotoxic
amino acid containing TG analogues from the
peptide prodrug by prostate-specific
proteases, such as
prostate-specific antigen and prostate-specific membrane
antigen, or
cancer-specific
proteases, such as fibroblast activation
protein, so that toxicity of these
prodrugs is selectively targeted to metastatic sites of
prostate cancer. Based on these results, these
prodrugs are undergoing clinical development.