A carboxymethylated-sulfated derivative of (1-->3)-beta-d-glucan (PCS3-II) extracted from Poria cocos was synthesized and coded as CS-PCS3-II. Results of infrared (IR) and Carbon-13 nuclear magnetic resonance spectroscopy ((13)C NMR) indicated that CS-PCS3-II contained carboxymethyl and sulfate groups with a degree of substitution (DS) of 1.05 and 0.36 respectively. By using size exclusion chromatography (SEC) combined with laser light scatting (LLS), the dependence of radius of gyration ((z)(1/2)) on the molecular weight (M(w)) for CS-PCS3-II was established as (z)(1/2) = 6.92 x 10(-2)M(w)(0.59) in 0.15M NaCl solution at 25 degrees C, suggesting that CS-PCS3-II existed as an extended flexible chain. CS-PCS3-II exhibited significantly higher inhibition ratio to Sarcoma 180 tumor in BALB/c mice than PCS3-II. Histological examination of tumor cells treated with CS-PCS3-II had signs of necrosis and apoptosis. It is postulated that introduction of the carboxymethyl and sulfate groups to PCS3-II increased its possible contact with the receptors of immune cells through hydrogen binding and electrostatic attraction, leading to a stronger immunological responses that resulted in inhibition of tumor cell proliferation. Moreover, there were significant increases in phagocyte and thymus indexes, spleen index, hemolytic activity as well as spleen antibody production and delayed type hypersensitivity (DTH), suggesting that CS-PCS3-II could significantly enhance immunpotentiation in mice.
