High-performance　near-infrared　(NIR)　probes　can　achieve　rapid　and　effective　imaging　and　tracking　of　tumor　biomarkers,　which　have　attracted　extensive　attention　in　tumor　visualization　and　early　diagnosis.　Fast　response　speed　and　high　contrast　fluorogenic　enhancement　are　important　criteria　for　high-performance　imaging　probes.　Here,　by　introducing　two　hydrophilic　sulfonates　into　the　probe　structure,　we　designed　and　optimized　a　new　NAD　(P)H:quinone　oxidoreductase-1　(NQO1)　activated　NIR　fluorogenic　probe　(S-QCy7-NQO1)　for　sensitive　detection　and　evaluation　of　NQO1　activity　in　cell　and　tumor　xenografts　mice.　The　catalytic　reduction　activity　of　NQO1　quickly　eliminated　the　highly　selective　quinone　trigger,　and　then　the　probe　S-QCy7-NQO1　was　converted　into　SQCy7　with　an　extended　conjugated　system,　resulting　in　a　245-fold　increase　in　NIR　fluorescence,　4.9　times　higher　than　that　of　QCy7-NQO1　without　sulfonates.　The　introduction　of　two　hydrophilic　sulfonates　also　endows　S-QCy7NQO1　a　fast　response　to　NQO1,　which　is　2　times　faster　than　QCy7-NQO1.　The　probe　S-QCy7-NQO1　showed　remarkable　responsiveness,　sensitivity　and　optical　performance　to　NQO1　under　physiological　conditions,　allowing　the　detection　of　endogenous　NQO1　activity　in　living　cells　with　an　ultra-high　positive-to-negative　ratio　(108-fold).　Notably,　with　the　improved　responsiveness　and　optical　properties,　it　was　further　applied　to　high　contrast　imaging　of　tumor-bearing　mice,　achieving　the　maximum　fluorescence　response　within　10　min.　On　this　basis,　S-QCy7-NQO1　can　be　a　potentially　practical　tool　for　early　cancer　diagnosis　characterized　by　elevated　NOQ1　levels.