Abstract: Most of human tumors over-express CXCR4. AMD3100, a nonpeptide antagonist for CXCR4 receptor, can be used for therapy of those tumors. It was found that metal ion complex, such as Cu²⁺, with AMD3100 enhanced its binding affinity to the receptor 10-fold higher as compared to AMD3100 alone. AMD3100 was synthesis from 3-aminopropyl ethylene diamine. ⁹⁹Tc^m-AMD3100 was labeled directly. Biodistribution studies were carried out in NH mice. SPECT imaging was performed in Hep-G2 tumor bearing mouse. The synthetic yield was 5.8% from 3-aminopropyl ethylene diamine to AMD3100. The labeling yield of ⁹⁹Tc^m-AMD3100 was over 98%. Biodistribution studies showed high accumulation of radiotracer in liver which had high-expression of CXCR4. SPECT imaging results showed that uptake in Hep-G2 tumor was high. The results showed that ⁹⁹Tc^m-AMD3100 was an attractive candidate for further development of SPECT radiotracer potentially suitable for CXCR4.