The objective of this study was to determine the response rate in chemonaive patients with inoperable, locally advanced, recurrent, or metastatic cervical cancer treated with pemetrexed (Alimta®, LY231514), a multitargeted antifolate. The patients were treated with either 500 mg/m2 (11 patients) or 600 mg/m2 (24 patients) of pemetrexed, administered as a 10-min infusion on day 1 of a 21-day cycle. Patients receiving 500 mg/m2 dose also received 5 mg/day oral folic acid supplementation beginning 2 days prior and ending on day 3 of each cycle. Of the 34 patients evaluable for efficacy, six patients (18%) had partial response, with median response duration of 3.8 months (range, 3.3–6.6 months). Twenty-four patients (71%) had stable disease, one patient (3%) had progressive disease, and three patients could not be assessed. Median overall survival was 15.2 months (range, 2.9–35.3+ months). Grade 4 hematologic toxicities consisted of neutropenia (37%), leukopenia (9%), anemia (6%), and thrombocytopenia (3%). One patient died of hypotensive shock associated with frank rectal hemorrhage that was considered to be related to the study drug. We conclude that pemetrexed therapy showed moderate activity, similar to other active agents, in the treatment of locally advanced or metastatic cervical cancer.