The present invention discloses thymopentin modified by heterocyclic carboxylic acid represented by general formula I, wherein RCO is 1,2,3,4-tetrahydro-β-carboline-3-formyl, β-carboline ‑3‑formyl, 1,2,3,4‑tetrahydroisoquinoline‑3‑formyl, isoquinoline‑3‑formyl, 1‑methyl‑1,2,3,4‑tetrahydro‑ β-carboline-3-formyl, 1-methyl-β-carboline-3-formyl and 4,5,6,7-tetrahydro-3H-imidazo[4,5-C]pyridine-6- Formyl, disclosed their preparation method, disclosed their anti-tumor cell proliferation activity in vitro, further disclosed their activity of inhibiting tumor weight gain in S180 tumor mice, and clarified their application in the preparation of anti-tumor drugs. RCO‑Arg‑Lys‑Asp‑Val‑Tyr I.