Background: Patients with mCRC are typically offered chemotherapy and might also receive target therapy-drugs targeting VEGF or EGFR. In phase 3 FRESCO trial, fruquintinib demonstrated a statistically significant and clinically meaningful overall survival benefit in third-line mCRC patients in China. To explore possible effects of prior target therapy on fruquintinib, we conducted subgroup analysis of patients with prior target therapy (PTT) and those without (non-PTT). Methods: Overall survival (OS) and progression-free survival (PFS) were evaluated by Kaplan-Meier method. Hazard ratio (HR) was estimated through Cox proportional hazards model. P-value was generated from log rank test. Results: Among a total of 278 fruquintinib-treated patients, 111 received prior target therapy (84 with anti-VEGF;40 with anti-EGFR and 13 with both). In PTT subgroup, fruquintinib significantly prolonged OS (Median OS: 7.69 months (m) vs 5.98 m;HR = 0.63;p = 0.023) and PFS (Median PFS:3.65m vs 1.84m;HR = 0.24;p < 0.001) compared to placebo. Patients who received prior anti-VEGF treatment (N = 84) also benefited from fruquintinib in OS (HR = 0.68,95%CI: 0.45-1.03) and PFS (HR = 0.24,95%CI: 0.15-0.38). In non-PTT subgroup, the median OS was 10.35 m for fruquintinib vs 6.93 m for placebo (HR = 0.63,p = 0.01) and the median PFS for fruquintinib was 3.81 m vs 1.84 m for placebo (HR = 0.28,p < 0.001). There were no observed accumulative Grade≥3 treatment-emergent adverse events (TEAEs) in PTT subgroup. Grade≥3 TEAEs rates of fruquintinib were similar in PTT and non-PTT subgroup (61.3% and 61.1%). The most common drug-related Grade ≥3 TEAEs of fruquintinib in PTT and non-PTT subgroup were hypertension (20.7% and 21.6%), hand-foot-skin reaction (7.2% and 13.2%) and proteinuria (5.4% and 1.8%). Conclusions: This subgroup analysis result is consistent with previously reported FRESCO intent-to-treatment population result. Fruquintinib showed clinically meaningful benefits in third-line mCRC patients regardless of prior target therapy without observed accumulative toxicity. Clinical trial information: NCT02314819