As a potential cancer immunotherapeutic agent, CHA has entered phase II clinical trials in China as a lyophilized powder formulation for treating glioma. However, the in vivo instability of CHA necessitates daily intramuscular injections, resulting in patient noncompliance. In this study, CHA-phospholipid complex (PC)-containing PEGylated liposomes (CHA-PC PEG-Lipo, named as CPPL), with CHA-PC as the drug intermediate, were prepared to lower the administration frequency. CPPL demonstrated excellent physicochemical properties, enhanced tumor accumulation, and inhibited tumor growth even when the administration interval was prolonged to 4 days when compared to a CHA solution and CHA-PC loaded liposomes (CHA-PC Lipo, labeled as CPL), both of which only demonstrated antitumor efficacy with once-daily administration.

These findings indicate that CHA-encapsulated mannosylated liposomes have great potential to enhance the immunotherapeutic efficacy of CHA by inducing a shift from the M2 to the M1 phenotype.

This review summarizes the pharmacological effects as well as the underlying mechanisms of CHA, providing an important theoretical basis for further research on CHA drug targets and potential mechanisms.

In this study, we confirmed the inhibition of proliferation by CHA in ESCC cells, as well as the reduction of ESCC xenograft volume by CHA in vivo. In addition, CHA also suppressed both the migration and invasion of ESCC cells in vitro. In a carcinogen-induced murine model of ESCC, hyperplasia of the esophagus was slowed by CHA, while mice suffering from ESCC that were treated with CHA had longer survival times than mice in the control group. The measurement of pluripotency factors (BMI1, SOX2, OCT4 and Nanog) that are related to poor prognosis revealed reduced expression of both BMI1 and SOX2, but not of OCT4 or Nanog, in ESCC cells, in both a dose- and time-dependent manner.

CHA might be a safe and effective differentiation-inducer for cancer therapy. “Educating”cancer cells to differentiate, rather than killing them, could be a novel therapeutic strategy for cancer.

A simple and specific, rapid resolution liquid chromatography-tandem mass spectrometry (LC–MS/MS) method was developed and validated for determination of CHA in human plasma using neochlorogenic acid as the internal standard. Plasma samples were precipitated with methanol and separated on a Zorbax C18 column (50×2.1 mm, i.d. 1.8 m) at a flow rate of 0.4 mL/min using a gradient mobile phase of methanol-water containing 0.1% formic acid (v/v). The detection was performed on a triple quadrupole tandem mass spectrometer by multiple reaction monitoring in negative ESI mode. The method was fully validated over the concentration range of 10–2000 ng/mL. The indicators of interand intra-day precision (RSD%) were all within 10.7%, and the accuracy (RE%) was ranged from -3.0% to 10.6%. Moreover, we evaluated this bioanalytical method by re-analysis of incurred samples as an additional measure of assay reproducibility. This method was successfully applied to pharmacokinetic study of CHA in Chinese subjects with advanced solid tumor after intramuscular injection administration of CHA for injection (CAFI).