Multi-drug-resistant (MDR) strains of biofilm-producing Escherichia coli are being reported worldwide and are threatening the health of human beings. These species are seen as the highest priority for the development of new phage biocontrol agents. Therefore, we isolated and characterized new and effective lytic phages as biocontrol agents against MDR E. coli. Bacteriophages (phages) were isolated from different wastewater samples, whilst E. coli strain was isolated from Sausage with accession number MN493640; it was resistant to 66.6% of tested common antibiotics with strong biofilm production. Phages designated ΦEcp1, ΦEcp2, ΦEcp3, ΦEcp4 and they were classified as being in the order Caudovirales, ΦEcp1 and ΦEcp2 belong to families Siphoviridae and Myoviridae respectively while ΦEcp3, ΦEcp4 belonging to family Podoviridae. The four phages and its cocktail showed polyvalent infectivity. All phages are stable at pH values range from 7–9; exhibited remarkable temperature stability (55-65ºC). They were stable in different concentrations of sodium chloride and did not lose their infectivity after exposure to Ultraviolet (UV) radiation for 30 min. One-step growth curve analysis showed short latent period (10 min) and large burst sizes (295, 117, 102 .and 127 PFU/mL) for ΦEcp1, ΦEcp2, ΦEcp3, ΦEcp4 respectively. Phages cocktail completely inhibited the growth of MDR E. coli in 8 h in vitro and also significantly (p < 0.05) eradicated the development of 24 h old biofilms. This study suggests that these four new phages can be characterized even farther as antimicrobial agents against biofilm-producing MDR E. coli.