The present invention relates to a thiophosphoryl amino acid ester compound. Said invention provides its structural formula, and its preparation method incldues the following steps: dissolving raw material (isopropoxy substituted) dichlorthion in tetrahydrofuran, adding amino acid methyl ester hydrochlorate to said solution, drop-adding acid-binding agent, and adding 2',3'-O-isopropylidene nucleoside solution in said solution, then more drop-adding acid-binding to make reaction, after reaction filtering, rotating distrillation, column chromatography to make separation, so that the invented compound can be obtained.

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