OBJECTIVE To explore whether the 12 Q-markers(6 anti-tumor and 6 cardiotoxic protective) selected by the research group from Aidi injection(AD) in the early stage have representative therapeutic effects on the two original formulas for AD.

METHODS Based on the preliminary work, the Q-markers with these two functions were formulated into different anti-tumor Q-marker compositions and various cardiotoxic protective Q-marker compositions, using the same concentration as the original AD formula. In comparison to the original AD formula, the anti-tumor effects of the compositions were evaluated on Huh7, HUVEC, and H9c2 cells using the conventional CCK8 method, as well as their cardiotoxic protective effects. Additionally, UHPLC-Q-Exactive Plus Orbitrap HRMS was employed for cellular metabolomics research to analyze the cellular metabolic profiles. The average distance value was calculated using the PLS-DA model to assess the similarity between the anti-tumor Q-marker compositions and the original AD formula, while the relative average distance value was calculated to evaluate the similarity of the cardiotoxic protective Q-marker compositions to the original AD formula.

RESULTS The results from the CCK8 assay and cellular metabolomics experiments were largely consistent, indicating that the combination of five anti-tumor Q-markers(cantharidin, ginsenoside Re, ginsenoside Rb1, astragaloside Ⅱ, and ginsenoside Rg2) outperformed the combination of six anti-tumor Q-markers and the original AD formula. Additionally, the five Q-markers with cardioprotective effects(eleutheroside E, syringin, ginsenoside Rd, isoflavone glycoside, and nonanedioic acid) also demonstrated superior efficacy compared to the six Q-markers combination and the original AD formula.

CONCLUSION The compounds cantharidin, ginsenoside Re, ginsenoside Rb1, astragaloside Ⅱ, and ginsenoside Rg2 may serve as potential Q-markers for the anti-tumor effects in AD. Additionally, eleutheroside E, syringin, ginsenoside Rd, isoflavone glucoside, and nonanedioic acid can act as potential Q-markers for the cardiotoxic protective effects against AD. These compounds exhibit comparable or superior anti-tumor and cardiotoxic protective efficacy compared to the original formulation.