Antitumor effects of hydrogen peroxide in vivo

Abstract

Glucose oxidase, covalently coupled to polystyrene microspheres (GOL), produced H(2)0(2) at an average rate of 3.6 nmol/min per 10(9) beads under standard assay conditions. Injection of 1.3 × 10(10) to 1.1 × 10(11) GOL i.p. prolonged the survival of mice by 27 percent after injection of 10(6) P388 lymphoma cells in the same site, consistent with destruction of 97.6 percent of the tumor cells. Placing mice for several hours in 100 percent O(2), the probable rate-limiting substrate for GOL, afforded a 42 percent prolongation of survival from P388 lymphoma, consistent with destruction of 99.6 percent of the tumor cells. When the P388 inoculum was 10(5), 10(4), or 10(3) cells, GOL led to long-term survival (presumed cure) of 23 percent, 77 percent, and 92 percent of the mice, respectively, consistent with reduction of the injected tumor dose to less than 10 cells. Subcutaneous growth of 10(5) P388 cells (approximately 300 lethal dose to 50 percent of mice) was suppressed in 83 percent of mice by admixture of GOL with the tumor cell inoculum. GOL alone had no effect against a more peroxide-resistant tumor, P815 mastocytoma. However, P815 cell glutathione reductase could be inhibited in vivo by well-tolerated doses of the antitumor agent, 1,3-bis(2-chloroethyl)- 1-nitrosourea (BCNU). BCNU alone cured few mice with P815. Together, BCNU and GOL apparently cured 86 percent of mice injected with 10(6) P815 cells i.p. The protective effect of GOL was abolished by boiling it to inactivate the enzyme, by co-injection of catalase coupled to latex beads, or by delaying the injection of tumor cells for 3 h, by which time the beads had formed aggregates. Soluble glucose oxidase, in doses threefold higher than that bound to GOL, had no detectable antitumor effect. A single injection of preformed H(2)0(2) readily killed P388 cells in the peritoneal cavity, but only at doses nearly lethal to the mice. In contrast, GOL had very little toxicity, as judged by the normal appearance of the mice for over 400 d, gross and microscopic findings at autopsy, and various blood tests. GOL injected i.p. remained in the peritoneal cavity, where it was gradually organized into granulomata by macrophages, without generalized inflammation. Thus, an H(2)0(2)-generating system confined to the tumor bed exerted clear- cut antitumor effects with little toxicity to the host.

Triggering of respiratory burst by phagocytosis in monocytes of patients with systemic lupus erythematosus

“…UK), i.e. 0 2 fim mean diameter polystyrene latex, was coupled with 320 fig rabbit IgG (Human Institute for SerobactcHological Production and Research, Budapest, Hungary) using 1-25 ml ofl'Vn l-(3-diethyIaminopropyl)-3-ethyI earbodimide hydrochloride {Sigma, Pootc, UK) as coupling agent [9]. The IgG-latex was resuspended in 0-02 M sodium phosphate buffer (pH 7-4) which eontains: 8 mg NaCl, 10 mg/ml BSA (Sigma).…”

Oxidation therapy: the use of a reactive oxygen species-generating enzyme system for tumour treatment

“…Previous studies using direct injections of hydrogen peroxidase into solid tumours were unsuccessful (Green & Westrop, 1958). A study using glucose oxidase conjugated to latex microspheres did not report attempts at treating solid tumours but rather tumour cells in the peritoneal cavity and co-injection of tumour cells with the micropheres at the time of s.c. implantation (Nathan & Cohn, 1981). However, modification of GO and PEG resulted in only a 40% reduction in activity as compared with a 90% loss of enzyme activity when conjugated to latex microspheres (Nathan & Cohn, 1981 Fischer 344 rat on day 0.…”

“…A study using glucose oxidase conjugated to latex microspheres did not report attempts at treating solid tumours but rather tumour cells in the peritoneal cavity and co-injection of tumour cells with the micropheres at the time of s.c. implantation (Nathan & Cohn, 1981). However, modification of GO and PEG resulted in only a 40% reduction in activity as compared with a 90% loss of enzyme activity when conjugated to latex microspheres (Nathan & Cohn, 1981 Fischer 344 rat on day 0. PEG-glucose oxidase and heatdenatured PEG-glucose oxidase was administered intratumorally on day 12 (noted by upward pointing arrow) at two separate doses of 200 U 3 h apart.…”

“…It was not until the early 1980s that investigators reported that hydrogen peroxide can exert a direct anti-tumour effect in vivo and thus prolong host survival (Nathan & Cohn, 1981). In that previous study, glucose oxidase (EC 1.1.3.4) was conjugated to carboxylated latex micropheres and was found to have an anti-tumour effect both in the peritoneal cavity and when injected into the tumour bed of subcutaneously implanted P388 cells at the time of cell implantation (Nathan & Cohn, 1981). The microspheres were at the tumour site and hydrogen peroxide generation was achieved through the following reaction: GO P-D-Glucose + 02------------> D-glucono-1,5-lactone + H202…”

Zanvil Alexander Cohn 1926-1993.

“…Tumor cell killing was manifest only in appropriately activated macrophages, especially those obtained from BCG-and C. parvum-primed mice. Tumor cell antiox- idant defenses could be compromised in vitro and in vivo (164,188,199,220,252), and this led to enhanced killing. In other interesting experiments, the group induced killing of targets, by applying peroxidase and a halide to cells (81), and sensitized organisms with peroxidase to small amounts of H202 (206).…”