The proposes of this research are to improve the delivery efficiency of AuNPs. One of the problems with drug loading and drug degradation, the drug was loaded onto the surface of the gold particles, adheres to the outer of the surface, which was easy to decompose in the body. In this work, we have developed the coated AuNPs by modified chitosan (Bi-QCS), which the Biotin-Quat188-AuNPs@collagen (Bi-QCS-AuNPS@collagen) was designed as follow: (i) collagen was used as a reducing agent and a stabilizing agent in the AuNPs@collagen, while (ii) modified chitosan (positive charge polymer) was coated on the outer layer, which modified chitosan (Bi-QCS) was consists of a positive charge (Quat188) and a targeting molecule (biotin) to increase cellular uptake and control drug release. After that, 5-Fluorouracil (5-FU) as an anticancer agent was loaded into BiQCS-AuNPs@collagen. The Bi-QCS-AuNPs@collagen was characterized by many techniques. The particle size of AuNPs@collagen increased after coating with Bi-QCS from 30 nm to 50 nm, while the zeta potential change from a negative value (uncoated) to a positive value (coated with Bi-QCS). The coating of Bi-QCS on the surface of AuNPs@collagen can increase the loading efficiency was 87.46 %, while the AuNP@collagen shows a loading efficiency of only 64.67 %. In addition, Bi-QCS-AuNPs@collagen can control the drug release rate, which at pH 5.4 showed the drug release rate higher than pH 7.4 after 48 hours. The evaluation of the biological activity of Bi-QCS-AuNPs@collagen presented significantly higher anti-inflammatory activity and anticancer activity of 5-FU.