Traditional pneumatic tyres are prone to puncture or blowout and other safety hazards. Non-pneumatic tyres use a high-strength, high-toughness support structure to replace the “airbag body” structure of pneumatic tyres, which is made of fibre skeleton materials and rubber laminated layers, thus effectively avoiding the problems of blowout and air leakage. However, discontinuous spokes undergo repeated bending deformation when carrying loads, which leads to energy loss, of which the rolling resistance of non-pneumatic tyres is one of the main sources of energy loss. This paper focuses on the study of gradient honeycomb non-pneumatic tyres. Firstly, a finite element model was established, and the accuracy of the model was verified by numerical simulation and stiffness tests. Secondly, the order of the effect of different spoke thicknesses on rolling resistance was obtained through orthogonal test analysis of four-layer honeycomb spoke thicknesses. Then, four optimized design variables were selected in combination with the spoke angles, and the effects of the design variables on rolling resistance were analyzed in detail by means of the Latin hypercube experimental design. Finally, the response surface model was established, and the non-linear optimization model was solved by the EVOL optimization algorithm considering the tyre stiffness limitations so that the rolling resistance was minimized. The results of the study laid down theoretical and methodological guidance for the design concept and technological innovation of low rolling resistance comfort non-pneumatic tyres.