Saussurea costus (S. costus) is an Indian medicine utilized to treat several diseases. It has a variety of pharmacological properties, including antioxidant activity, and contains bioactive compounds. The goal of this study is to explore the antioxidant activity, antimicrobial, cytotoxic, and anti-Alzheimer's activity of S. costus. According to the results, S. costus root powder recorded the following contents: moisture, ash, protein, fat, and available carbohydrates, which were 3.90, 4.38, 6.60, 5.98, and 84.04 %, respectively. Total flavonoids were detected in greater quantities than total phenol (161.20 mg QC /100g and 254.86 mg GAE/100g, respectively), and the DPPH scavenging radical value was 88.84%. HPLC is used to fractionate the diversity and concentra-tion of the polyphenolic compounds. The main flavonoid components were naringin (14.58 mg/100g), while ellagic acid was the primary phenolic acid (14.83 mg/100g). S. costus was highly effective as an antimicrobial due to the presence of these components. In addition, the root powder exhibited minimal toxicity against the Vero cell, with an IC50 of 70.50±3.17 μg/ml value. The results demonstrated that the inhibition percentage of the acetylcholinesterase (AChE) enzyme associated with Alzheimer's disease increased with higher concentrations of S. costus root extract, which exhibited IC50 of 155.3±10.28 μg/ml compared to the control (Donepezil standard). Partial substitution of wheat flour with 1 to 4% S. costus powder enhanced the antioxidant activity of crackers by increasing polyphenolic compound content and nutritional value while maintaining sensory quality and con-sumer acceptability. Furthermore, the low total microbiological count observed in the cracker formulations dur-ing the three-month storage period highlights the natural antioxidant and antimicrobial efficacy of S. costus powder. Given the complex, multifactorial etiology of Alzheimer's disease, this root powder represents a safe and health-promoting candidate for the future increase of disease-modifying therapies targeting Alzheimer's.