Initiating the system using 4 (Figure ), we found that during the time-course of the reaction cascade by 1 H NMR (Supplementary Figure ), compound 4 exhibits a sigmoidal amplification profile, which is a typical attribute for auto-amplification systems. In particular, the use of 0.67 mM SH-signal (~5%) to initiate the reaction resulted in thiol concentration increase to approximately 14.3 mM (~105%) after 45 hours, accounting for full conversion of 1 (Figure &). Matching this process, disulfide concentration quantitatively decreases ~13.5 mM from 20.3 to 7.4 mM. Simulations using the kinetic model for 3 (R 2 = 0.93) and 4 were consistent with the experimental data. Importantly, the model was able to describe the sigmoidal amplification profile of 4 in detail (Figure insert) and with high accuracy (R 2 = 0.96). Furthermore, we found that the production of 2 corresponds to the conversion of 1 (Figure -middle). Here, our simulations substantiate the exponential decrease of 1 and increase of 2, with R 2 -values of 0.98 (1) and 0.96 (2). Similarly, OTPPTS concentration is in excellent agreement with simulation results (R 2 = 0.98) (Figure -bottom). Using 1 H NMR spectroscopy to track all species involved in the two-component reaction cycle and describing those with kinetic simulations, we were able to confirm successful amplification of thiol 4 though coupled nucleophilic substitution and disulfide reduction reactions.