Niosomes are self-organizing non-ionic surfactant vesicles, which encapsulate aqueous volume of drug(s) with or without the addition of cholesterol and other lipid contents. Niosomes have the capability to encapsulate both lipophilic and hydrophilic drugs. They are alternative to liposomes, and their main benefits as compared to liposomes are their lower price, higher stability and better biodegradability. By making niosomes, the side effects of drugs have been reduced and the therapeutic efficacy has been increased. The first part of the study was to develop an optimized niosome formulation for the encapsulation of a poorly water-soluble drug by the ecological probe sonication method. Pluronic L121 and Span 60 were used as surface active agents and the optimization of the composition was made with the aid of Design of Experiment (DoE) concept. Rifampicin was used as a model drug. Concentration levels of charge inducing agent, dicetylphosphate (DCP), and Pluronic L121 were studied as variables. Prepared niosomes with varying concentrations of DCP and Pluronic L121 resulted in small sized niosomes with sizes ranging from 190 nm to 893 nm. During the four weeks stability testing, the particle sizes were reduced slightly. The formulation containing 2 mg of DCP resulted in most stable niosomes with 75.37% entrapment efficiency. All the niosomal formulations showed higher In vitro drug release rates as compared to bulk drug formulation. The rifampicin loaded niosomes prepared with Pluronic L121 and Span 60 resulted in stable, small sized niosomes with improved drug release profile. The second part of the study was carried out to produce niosome formulations for the encapsulation of a hydrophilic and poorly water-soluble drugs by the ecological probe sonication method. Pluronic L121 and Span 60 were used as surface active agents and the optimization of the composition was made with the aid of Design of Experiment (DoE) concept. Ceftriaxone sodium and Rifampicin were used as model drugs (hydrophilic and hydrophobic respectively). Concentration levels of charge inducing agent, dicetylphosphate (DCP), and Pluronic L121 were studied as variables. Prepared niosomes with varying concentrations of DCP and Pluronic L121 resulted in small sized niosomes with sizes ranging from 164 nm to 893 nm. During the four weeks stability testing, the particle sizes were reduced slightly. The formulations CR1 and CR2 resulted in most stable niosomes with (98.71% of rifampicin and 95.73% ceftriaxone) and (98.86% rifampicin and 95.88% ceftriaxone) entrapment efficiency of respective formulations. All the niosomal formulations showed higher In vitro drug release rates as compared to bulk drug formulations. The ceftriaxone and rifampicin loaded niosomes prepared with Pluronic L121 and Span 60 resulted in stable, small sized niosomes with improved drug release profile. In the third part of study, the niosome formulations were prepared for the encapsulation of anticancer drugs by the ecological probe sonication method. Pluronic L121 and Span 60 were used as surface active agents in niosomes and doxorubicin HCl and paclitaxel were used as anticancer drugs. Thorough physicochemical studies were performed for the niosomes and their cytotoxicity and activity were evaluated on MCF-7 and PC3- MM2 cancerous cell lines. Prepared niosomes were small with sizes ranging from 137 nm to 893 nm and entrapment efficiencies were high, ranging from 91.24% to 99.99%. During the four weeks stability testing, the particle sizes remained stable. The niosomal formulations showed In vitro sustained drug release profiles for doxorubicin HCL and increased clearly dissolution rate of poorly water-soluble paclitaxel. The incorporation of both the drugs into niosomes, improved cell penetration and antiproliferative activity of the drugs towards PC3 as compared to MCF-7 cell lines. As a conclusion, doxorubicin HCl and paclitaxel loaded niosome formulations resulted in relatively stable, small sized niosomes with improved drug release profiles, better cell penetration and antiproliferative activity. The niosomes showed more antiproliferative effect due to the presence of both drugs, which can overcome multidrug resistance. The present study suggested that the codelivery of anticancer drugs can be delivered by encapsulating in niosomes prepared from Pluronic L121 and Span 60. Through which improved in-vitro sustained release of both anticancer drugs, better cell penetration and antiproliferative activity. The further in-vivo evaluation can lead to treat different types of cancers in a better way without toxic effects with reduction in doses.