Due to the emerging problem of multidrug resistant (MDR) pathogens, there is a demand of novel biological active compounds to replace currently used antibiotics in order to evade resistance. Currently, the pathogenic organisms of concern are methicillin and vancomycin resistant strains of Staphylococcus aureus, Acinetobacter spp., extended spectrum β-lactamases (ESBLs) producing strains of Escherichia coli, Klebsiella spp., Pseudomonas spp. and Proteus spp. In this study, a collection of nineteen actinomycetes strains isolated from Cholistan Desert, Pakistan, were screened biologically and biochemically for their potential bioactive secondary metabolites against MDR bacterial pathogens. The identification of strains was done by morphological, physiological and biochemical characterization and by 16SrRNA gene sequencing. For biological screening different methods including cross streak, agar plug and well diffusion method were performed. Seven Streptomyces strains including NWMD-6, NWMD-7, NWMD-10, NWMD-11, NWMD-12, NWMD-15 and NWMD-16 were found to have significant antimicrobial activity against MDR bacterial pathogens. Secondary metabolite profiling of each methanolic extract by TLC and HPLC-UV signified the presence of unique and diverse metabolic compounds. The study predicts that the seactinomycetes strains are a potential resource to cope up with evolving multi drug resistant pathogens. Furthermore, these secondary metabolites can be identified by comparative studies and these actinomycetes can be grown on larger scale to obtain high yield of potentially useful secondary metabolites.