Peter P. McCann

Matrix metalloproteinase inhibition as a novel anticancer strategy: a review with special focus on batimastat and marimastat.

Matrix metalloproteinases (MMPs) are a homologous family of enzymes that are involved in tissue remodeling and morphogenesis. Collectively, these enzymes are capable of degrading all components of the extracellular matrix, and they play an important role in normal physiologic conditions, such as wound healing and other processes involving tissue remodeling. However, increased activity of these enzymes now has been observed in a number of different pathological conditions, and it has been hypothesized that such increased activity of MMPs might play a role in the pathogenesis of these conditions. Cancer is one such condition; extracellular matrices constitute the principal barrier to tumor growth and spread, and there is growing experimental evidence that malignant tumors utilize MMPs to overcome these barriers. Consequently, inhibitors of MMPs represent an attractive target for a new class of anticancer agents. Marimastat and batimastat are potent broad-spectrum inhibitors of all major MMPs and have been shown to prevent or reduce spread and growth of a number of different malignant tumors in numerous animal models. Both agents are now in advanced clinical testing in a number of different solid tumors in North America and Europe. The purpose of this paper is to review available preclinical and emerging clinical data, using batimastat and marimastat as prototype MMP inhibitors in the cancer area.

Ornithine decarboxylase as an enzyme target for therapy

Interest in ornithine decarboxylase (ODC) and the therapeutic effects of its inhibition with the consequent depletion of polyamine biosynthesis has been widespread since the late 1970s and 1980s. This review covers new information about the properties of ODC, recent findings with ODC inhibitors and a discussion of the mechanism of inactivation of ODC by eflornithine. Recent in vivo therapeutic approaches of ODC inhibition are also discussed including: cancer and cancer chemoprevention; autoimmune diseases; polyamines and the blood-brain barrier, ischemia and hyperplasia; the NMDA receptor and modulation by polyamines; hearing loss; African trypanosomiasis; Pneumocystis carinii pneumonia and Cryptosporidium in AIDS; and other infectious diseases/organisms.

Biphasic induction of ornithine decarboxylase and putrescine levels in growing HTC cells.

Abstract A biphasic induction of ornithine decarboxylase with concomitant increase of intracellular putrescine was seen in growing rat hepatoma cells during each generation period. In non-growing HTC cells no coordinate accumulation of intracellular putrescine followed the unique induction of DDC by dilution into fresh serum-deprived medium. The data together suggest that the biphasic increases of ODC activity occur just before and after DNA synthesis and that a growing HTC cell has a finely regulated cycle of ODC activity. Finally, ODC activity may not always correlate with the intracellular putrescine levels.

Regulation of ornithine decarboxylase by ODC-antizyme in HTC cells.

Abstract Low concentrations of putrescine (10 −5 M) blocked ornithine decarboxylase (ODC) in rat hepatoma (HTC) cells in culture, but the lower homologue of putrescine, 1, 3 diaminopropane, had no effect on ornithine decarboxylase at 10 −5 M. Higher concentrations of both putrescine and 1, 3 diaminopropane induced approximately the same amount of soluble ODC antizyme type inhibitor. When concentrated dialyzed supernatants of cells grown in 10 −5 M putrescine were treated with 250 mM NaCl and chromatographed on a superfine Sephadex G-75 column, both ODC and inhibitor were recovered. Spermidine, spermine and cadaverine also induced the inhibitor suggesting a low specificity of induction by amines.

Effect of α-methyl ornithine on ornithine decarboxylase activity of rat hepatoma cells in culture

Abstract DL-α-methyl ornithine (α-MeOrn), a potent competitive inhibitor of ornithine decarboxylase (L-ornithine carboxy-lyase EC 4.1.1.7) (ODC), caused an increase of ODC activity of rat hepatoma tissue culture cells cultured in the presence of the compound. The effect was seen only when there was newly synthesized ODC present. The α-MeOrn mediated increase of ODC activity was independent of RNA synthesis and was expressed at a post-transcriptional level. ODC half-life, measured after cycloheximide treatment, was increased by a factor of three in the presence of α-MeOrn, suggesting the compound slowed enzyme degradation perhaps by its ability to enter the active site.

Inhibition of bacterial aminopropyltransferases by S-adenosyl-1,8-diamino-3-thiooctane and by dicyclohexylamine

Bacterial aminopropyltransferases from Escherichia coli, Serratia marcescens and Pseudomonas aeruginosa were strongly inhibited by S‐adenosyl‐1,8‐diamino‐3‐thiooctane (AdoDATO) and by dicyclohexylamine. The sensitivity to these drugs in vitro was comparable to that of mammalian spermidine synthase, but AdoDATO was much less potent in reducing spermidine content in the bacteria than in mammalian cells. Although AdoDATO was a stronger inhibitor than dicyclohexylamine in vitro, dicyclohexylamine was more active in reducing bacterial spermidine levels in vivo, suggesting that it is take up better or is more stable in the cell and is the preferable compound for in vivo studies in microorganisms. The strong inhibition of spermidine synthases by AdoDATO which is a transition state analog supports the concept that these enzymes proceed by a single displacement reaction, rather than by a ping‐pong mechanism.

Regulatory interrelations between GABA and polyamines. II. Effect of GABA on ornithine decarboxylase and putrescine levels in cell culture

GABA added to rat hepatoma (HTC) cells in spinner culture at the time of induction of cell proliferation increased levels of ornithine decarboxylase (ODC) up to two- to threefold above that of control cells. The increases in ODC were also reflected by concomitant increases of intracellular putrescine levels, while spermidine and spermine were unchanged. GABA seems to have a direct stabilizing effect on ODC, since the turnover of the enzyme was slowed almost twofold when measured in cells treated with 10−2 M GABA. The stabilizing effect is most pronounced for GABA, although some amino acids such as asparagine, glutamine, and lysine as well as some GABA analogues and homologues also tend to increase ODC but to a significantly lesser extent than GABA itself. GABA metabolites had no effect on ODC.S-Adenosylmethionine decarboxylase and tyrosine aminotransferase were not affected by the presence of GABA. The GABA effect on ODC may be important in certain types of cells for the regulation of polyamine biosynthesis.

The dual action of the non-physiological diamines 1,3 diaminopropane and cadaverine on ornithine decarboxylase of HTC cells

Abstract In rat hepatoma tumor (HTC) cells 1,3 diaminopropane and cadaverine induced the ornithine decarboxylase antizyme as well as the end product of the ornithine decarboxylase reaction putrescine. Although at equal exogenous concentrations (10−3M) the two non-physiological diamines penetrated the cells as effectively as putrescine; they decreased cellular ornithine decarboxylase considerably less rapidly than the naturally present diamine. Cell extracts treated with high concentrations of 1,3 diaminopropane and putrescine, and which as a result had a high specific activity of ornithine decarboxylase antizyme, were chromatographed on a superfine Sephadex G-75 column in the presence of 250 mM NaCl. No ornithine decarboxylase-antizyme complex could be detected indicating the original decrease of ornithine decarboxylase in the cells was likely due to some mechanism other than antizyme. These results indicate that 1,3 diaminopropane and cadaverine probably can act on ornithine decarboxylase, like putrescine, by two distinct regulatory mechanisms.