Objective: Paclitaxel is one of the most effective anticancer agents. It is used as a chemotherapy agent for a spectrum of cancer types. However, paclitaxel resistance is one of the foremost problems for chemotherapy. Most importantly, an emergence of paclitaxel resistance due to mutation (F270V) in β-tubulin has been extremely deliberated in recent years. With the rise of paclitaxel-resistant mutation in β-tubulin, there is a need to add a novel inhibitor from natural source, as they have less chance of getting resistance additionally less side effects. Keeping this in mind, we have utilized experimental and in silico approaches to isolate the potent inhibitor for β-tubulin target protein. Methods: We have extracted phytocompounds from Cassia fistula plant, and the structures were recognized with the help of gas chromatographymass spectrometry technique. Subsequently, oral bioavailability and toxicity analysis were executed for the extracted compounds by employing MOLINSPIRATION and OSIRIS program, respectively. Furthermore, docking analysis was performed using YASARA algorithm. In addition, bioactivity analysis for the screened compounds was performed using prediction of activity spectra for substances program. Results: The results from our analysis clearly depict that HOP-22(29)-EN-3.BETA.-OL could be a promising inhibitor for the treatment of cancer and provide direction for future research. Further in vitro and in vivo exploration is also required to identify whether HOP-22(29)-EN-3.BETA.-OL have anticancer effect or not. Conclusion: The combination of computational approach and experimental analysis provides an easy approach to identify novel candidate for the target protein β-tubulin. © 2016, Innovare Academics Sciences Pvt. Ltd. All rights reserved.