Abstract Background Hypoxia-ischemic encephalopathy (HIE) is one of the most important complications of the newborn period. It is the result of depriving nerve tissue of oxygen and glucose, which may be the result of either hypoxia or ischemia. Objective To assess effect of therapeutic hypothermia (TH) on EEG changes and Carboxy-terminal hydrolase L1level in neonatal HIE at Al-Zahraa university and El Haram hospitals. Patients and Methods This was a prospective comparative study that was conducted at the NICU unit Al ZahraaUniversity and El Haram hospitals for a period from (May 2020 till June 2022). This study included 50 full term newborns (37 to 40 weeks of gestational age) with clinical and laboratory evidence of HIE II–III, They were divided into two groups: Group 1: 25 HIE neonates who were treated with therapeutic hypothermia and group 2: 25 HIE neonates who weren’t received therapeutic hypothermia. Newborns with congenital anomalies, intracranial lesion, epileptic seizure, spinal cord injury or cardiac arrest were excluded from the study. We record EEG changes during 72 hrs by an electroencephalogram with a bioelectric amplifier (Deltamed, Bourgogne), (Philips) Screen. Blood samples were analyzed for UCH-L1 levels by (SEG945Mi). The neonates involved in cooling group were placed over a cooling blanket (Blanketrol III), for 72 hours. Results The results showed there was statistically significant relation between high UCH-L1 level and higher grades of the Sarnat score and primitive reflexes of both the cooling group and non cooling group. The results also showed that there were lower levels of UCH-L1 in cooling group compared to non cooling group of the studied neonates Also there was significant negative correlation found between UCH-L1 level and PH, and HCO3 values. This study showed that there was statistically significant relation found between UCH-L1 level and EEG recording on the 3rd day of both the studied groups. This study showed that (84%) of the non cooling group had EEG abnormalities compared to (68%) of the cooling group. Among both groups of the studied neonates, 76% had EEG abnormalities in form of (34%) had suppression EEG, (16%) were occipito-temporal, (20%) fronto-temporal and (6%) centro-parital. Regarding the site of EEG abnormality (18%) were right sided, (24%) were left sided. The study showed that 72% of cooling group were discharged compared to 48% of non cooling group. Conclusion From our study we concluded significant improvement in EEG records in ischemic neonates who were treated with therapeutic hypothermia. UCH- L1 is a good early neuronal injury biomarker for detecting severity of HIE P value < 0.001 and is a good predictor of hypoxia with specificity 92% and ppv 86.7% and NPV 65.7%. Therapeutic hypothermia has been noted to reduce severe disability among survivors.