In this paper, we introduce and investigate a new subclass of bi-prestarlike functions defined in the open unit disk, associated with Chebyshev Polynomials. Furthermore, we find estimates of first two coefficients of functions in these classes, making use of the Chebyshev polynomials. Also, we obtain the Fekete-Szegö inequalities for function in these classes. Several consequences of the results are also pointed out as corollaries.