Small molecule protein assembly modulators with Pan-Cancer therapeutic efficacy

Two structurally-unrelated small molecule chemotypes, represented by compounds PAV-617 and PAV-951 with antiviral activity in cell culture against Mpox virus (Formerly known as monkeypox virus) and human immunodeficiency virus (HIV) respectively, were studied for anti-cancer efficacy. Each exhibited apparent pan-cancer cytotoxicity, reasonable pharmacokinetics, and non-toxicity in mice at active concentrations. Anti-tumor properties of both chemotypes were validated in mouse xenografts against A549 human lung cancer and, for one of the chemotypes against HT-29 colorectal cancer. The targets of these compounds are unconventional: each binds to a different transient, energy-dependent multi-protein complex. Treatment with these compounds alters the target multi-protein complexes in a manner that appears to remove a block, crucial for cancer survival and progression, on a homeostatic linkage between uncontrolled proliferation and apoptosis. These compounds provide starting points for develo..., Chemical Synthesis
All compounds synthesized were confirmed by LCMS with purity typically > 98%
Synthesis of PAV-617 (see Supplemental Figure 1A)
Phenothiazin-5-ium tetraiodide hydrate
Phenothiazin-5-ium tetraiodide hydrate:  a solution of phenothiazine (4.98 g, 25 mmol) in anhydrous chloroform (50 mL) was stirred at 50C and the solution of iodine (12.7 g, 50 mmol) in CHCl3 (250 mL) was added dropwise over 4h. The resulting dark solution was stirred for an additional 3h at 50C, monitored by TLC. After the disappearance of the starting material, the resulting precipitate was filtered, washed with a copious amount of chloroform, dried overnight in vacuo to afford a dark solid (13.9 g, 74%).
3,7-Di(pyrrolidin-1-yl) phenothiazinium iodide
A solution of phenothiazin-5-ium tetraiodide hydrate (2.8 g, 3.6 mmol) in mixture acetonitrile/methanol (50 mL) and pyrrolidine (710 mg, 10 mmol) was stirred for    4 h at room temperature. The resulting mixture was concentrated to dryness and purified ..., , # Small Molecule Protein Assembly Modulators with Pan-Cancer Therapeutic Efficacy

[https://doi.org/10.5061/dryad.k3j9kd5hf](https://doi.org/10.5061/dryad.k3j9kd5hf)

**File description:**

Each tab in the Excel file contains the raw data corresponding to a specific figure. Tabs are labeled according to the figure number, with supplemental figures indicated by an 's' before the number.

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**Tab descriptions:**

**Figure 1**

1A- shows assessment of the endogenous apoptosis response in multiple cell lines. Plates were seeded with LNCaP C-33, LNCaP C-81, CHO K-1, and Hennes 20 cells. After three days of growth Apo-ONE reagent was added and caspase-3/7 activity was determined by fluorescent readout. Excel document contains F521 absorbance (RFU) values for each cell line.

1D and 1E- show activity of PAV-617 and PAV-951 in the Hennes 20 “arrest of proliferation” assay where parallel p...