The effect of the antitumorally active hexadecylphosphocholine (He-PC) on the colony-stimulating factor (CSF)-dependent growth of human hemopoietic progenitor cells was studied. At low concentrations He-PC stimulated the CSF-dependent progenitor cell colony growth of three patients suffering from chronic myeloid leukemia (CML) and of three of six patients without hematological disorders. The stimulating effect was up to eight times that of the control using granulocyte colony-stimulating factor (G-CSF) and twofold in the case of granulocyte-macrophage colony-stimulating factor (GM-CSF), whereas only slight effects were noted when interleukin 3 (IL-3) or the combination of the CSFs was used as an additive. The stimulatory effects observed are far below the He-PC concentrations that are usually required for the in vitro growth arrest of cancer cells. At higher concentrations He-PC displayed suppressive effects, most pronounced in the case of G-CSF-dependent colony growth. At the concentrations investigated, He-PC failed to show any changes in the composition and distribution of specific colonies. He-PC by itself had no mitogenic activity. This indicates that He-PC acts as a co-stimulator. In the cases of myeloproliferative diseases and in the case of a patient without known hematological disorder, removal of accessory cells did not abrogate the He-PC-enhanced colony growth by CSFs. Thus, the stimulatory effect of low-dose He-PC seems not to be mediated by accessory cells.

Aged↗

PubMed1991

Active-site-directed specific competitive inhibitors of phospholipase A2: novel transition-state analogues.

More than 100 amphiphilic phosphoesters, possible tetrahedral transition-state analogues capable of coordinating to the calcium ion at the active site of phospholipase A2, were designed, synthesized, and tested as inhibitors for the hydrolysis of 1,2-dimyristoyl-sn-glycero-3-phosphomethanol vesicles in the scooting mode. This assay system permits the study of structurally diverse inhibitors with phospholipase A2S from different sources, and it is not perturbed by factors that change the quality of the interface. As a prototype, 1-hexadecyl-3-trifluoroethylglycero-2-phosphomethanol (MJ33) was investigated in detail. Only the (S)-(+) analogue of MJ33 is inhibitory, and it is as effective as the sn-2 phosphonate or the sn-2 amide analogues of sn-3 phospholipids. The inhibitory potencies of the various phosphoesters depended strongly on the stereochemical and structural features, and the mole fractions of inhibitors required for 50% inhibition, X1(50), ranged from more than 1 to less than 0.001 mole fraction. The affinity of certain inhibitors for enzymes from different sources differed by more than 200-fold. The inhibitors protected the catalytic site residue His-48 from alkylation in the presence of calcium but not barium as expected if the formation of the EI complex is supported only by calcium. The equilibrium dissociation constant for the inhibitor bound to the enzyme at the interface was correlated with the XI(50) values, which were different if the inhibition was monitored in the pseudo-zero-order or the first-order region of the progress curve. These results show that the inhibitors described here interfered only with the catalytic turnover by phospholipase A2's bound to the interface, their binding to the enzyme occurred through calcium, and the inhibitors did not have any effect on the dissociation of the enzyme bound to the interface.

Animals↗

PubMed1991

Structural requirements of lysophospholipid-regulated mitochondrial Ca2+ transport.

Analogues of lysophosphatidylcholine, including PAF (platelet-activating-factor) and HePC (an experimental anticancer drug), were studied for their influence on mitochondrial Ca2+ transport and membrane potential. Lysophospholipids released Ca2+ from mitochondria and reduced the maximal Ca2+ uptake. The structure-activity relations indicate that deprotonated head groups like phosphocholines yield active compounds while partially protonated head groups like phosphoethanolamines are essentially inactive. Structural requirements for the apolar part of the molecules were acyl or alkyl chain lengths of less than 18 carbon atoms at the C1-position of the glycerol backbone and residues of small size and/or low polarity at the C2-position. Choline lysophospholipids, but not ethanolamine lysophospholipids, may therefore induce mitochondrial Ca2+ efflux and become mediators of ischaemic tissue damage where dysregulated phospholipase A2 activity and an impairment of mitochondrial function are supposed to play a crucial role.

Animals↗

PubMed1991

Hexadecylphosphocholine-mediated enhancement of T-cell responses to interleukin 2.

The effect of low-dose hexadecylphosphocholine (He-PC) on normal peripheral mononuclear cells (PMNC) was studied. Interferon-gamma (IFN-g) production, interleukin 2 (IL-2) receptor, and HLA-DR antigen expression were investigated, representing typical T-cell activation parameters. In PMNC cultures, He-PC dose-dependently enhanced the production of IFN-g, provided IL-2 had been added exogenously. Without IL-2 He-PC was ineffective. In some cultures, at a concentration of 8 micrograms/ml He-PC stimulated the secretion of IFN-g more than 20-fold compared to untreated controls. Although He-PC by itself lacked mitogenic activity, this compound also stimulated IFN-g production in the presence of suboptimal doses of phytohemagglutinin (PHA). Immunofluorescence studies demonstrated that He-PC also increased IL-2 receptor and HLA-DR antigen expression under these experimental conditions. Taken together, these results indicate that He-PC may possess immunomodulatory activity also in vivo, acting as a costimulator for the IL-2-mediated T-cell activation process.

Drug Synergism↗

PubMed1991

Hexadecylphosphocholine: preclinical and the first clinical results of a new antitumor drug.

Dose-response studies on cytotoxic alkyl lysophospholipids with various chemical structures revealed that a long alkyl chain and a polar group are essential for antitumor activity. The combination of both the long alkyl chain and a phosphocholine group thus results in alkyl phosphocholines. Preclinical studies with hexadecylphosphocholine (He-PC) as a representative compound indicate distinct antineoplastic activity on leukemia cells of human origin. He-PC is highly effective in inhibiting the growth of chemically induced rat mammary carcinomas. Even more striking is the fact that a high percentage of the tumors regressed completely. In a clinical phase I trial on breast cancer patients with local recurrences, topically applied He-PC resulted in regression of skin metastases. A phase II trial for topical treatment and a phase I trial for orally applied He-PC have been initiated to further evaluate the antitumoral activity of this new compound.

Adult↗

PubMed1991

Comparison of selective cytotoxicity of alkyl lysophospholipids.

Alkyl lysophospholipids have been shown to be cytooxic to a number of neoplastic tissues. One, ET-18-OCH3, has been used to selectively purge leukemic cells from mixtures with normal marrow progenitor cells, in vitro and in vivo. We have measured the 50% inhibitory (IC50) effect of a series of ether lipids (EL) on leukemic cells (HL60, K562, Daudi, KG-1, KG-1a) and normal marrow progenitor cells. Cells were incubated with varying concentrations of EL for 4 hr and assayed for viability, [3H]thymidine incorporation and clonogenicity in semi-solid media. The effect on protein kinase C (PKC) activity was assayed for each compound. Compounds tested included three glycerophosphocholine analogs--ET-18-OCH3, ET-16-NHCOCH3, and BM 41.440. In addition, a lipoidal amine, CP 46665, an ethyleneglycolphospholipid, AEPL, and four single chain alkylphosphocholine analogs, HePC2, HePC3, HePC4 and HePC6 were also tested. During the period of incubation, the cells remained viable (greater than 70%) as judged by trypan blue dye exclusion. The glycerophosphocholines were the most active and showed the highest therapeutic index. The lipoidal amine was active, but toxic to normal marrow progenitor cells. The ethyleneglycolphospholipid was active against HL60, but not against the other cell lines. The single chain alkylphosphocholine analogs were less active. All of the compounds inhibited PKC activity; however, the glycerophosphocholines were the most inhibitory.

Antineoplastic Agents↗

PubMed1991

Effects of hexadecylphosphocholine on protein kinase C and TPA-induced differentiation of HL60 cells.

Several structural analogs of alkylphosphocholine (APC) were studied for their effects on protein kinase C (PKC) and 12-O-tetradecanoylphorbol-13-acetate (TPA) elicited biochemical and cellular events in HL60 cells. Hexadecylphocholine (He-PC2), the APC prototype, inhibited PKC competitively with respect to phosphatidylserine an noncompetitively with respect to CaCl2, both with an apparent Ki of about 15 microM. Inhibition of PKC by He-PC2 was selective, since cyclic AMP dependent protein kinase and Ca2+/calmodulin dependent protein kinase II were relatively unaffected. He-PC2 inhibited TPA-induced depletion of PKC and TPA-stimulated phosphorylation of cellular proteins in HL60 cells. TPA-induced differentiation of HL60 cells was also inhibited by He-PC2, and this inhibition was synergistic or additive to the effects of 1-(5-isoquinolinesulfonyl)-2-methylpiperazine (H-7), a PKC inhibitor. The present findings are consistent with the hypothesis that inhibition of PKC might be related, in part, to the antineoplastic effect of He-PC2 and ether lipid analogs such as ET-18-OCH3 (1-octadecyl-2-methyl-glycero-3-phosphocholine).

Cell Differentiation↗

PubMed1990

Synthesis and use of a lysolecithin analog for the purification of UDP-glucuronosyltransferase.

Because of their high cost, lysolecithins are generally not considered useful detergents for the purification of membrane-bound enzymes. Therefore, we have synthesized a structural analog of lysolecithin with similar physical properties for which synthesis is straightforward. This analog is 1-palmitoylpropanediol-3-phosphocholine. To compare the efficacy of the two detergents for the purification of a membrane-bound enzyme, we have purified UDP-glucuronosyltransferase from pig liver microsomes using lysophosphatidylcholine or the synthetic analog. The catalytic properties of UDP-glucuronosyltransferase purified with 1-palmitoylpropanediol-3-phosphocholine or lysolecithin were identical. Sodium dodecyl sulfate-gel electrophoresis indicated that the purity of the UDP-glucuronosyltransferase preparation was the same whether lysophosphatidylcholine or its synthetic analog was used. The advantage of using 1-palmitoylpropanediol-3-phosphocholine in preference to lysophosphatidylcholine is that the former can be synthesized for about 1% the cost of the latter. In addition, the method for synthesis of 1-palmitoylpropanediol-3-phosphocholine is general in that the structural features of the polymethylene chain can be varied, allowing for the inexpensive synthesis of a series of detergents.

Electrophoresis, Polyacrylamide Gel↗

PubMed1990

Inhibition of protein kinase C, (sodium plus potassium)-activated adenosine triphosphatase, and sodium pump by synthetic phospholipid analogues.

The effects and modes of action of certain antineoplastic phospholipid analogues (racemic 1-O-octadecyl-2-O-methyl glycero-3-phosphocholine, BM 41.440, JH-1, CV-3988, and HePC) on (sodium plus potassium)-activated adenosine triphosphatase (Na,K-ATPase) and sodium pump activities were investigated. Inhibition of Na,K-ATPase in purified rat brain synaptosomal membranes by these lipids, in contrast to ouabain, was subject to membrane surface dilution and unaffected by whether the reaction was started with KCl, NaCl, or ATP. Kinetic analysis indicated that the analogues, again dissimilar to ouabain, were likely to interact directly or indirectly with sodium-binding sites of Na,K-ATPase located at the intracellular surface of the plasma membrane, a conclusion also supported by studies using the inside-out vesicles of human erythrocyte membranes. The studies also showed that ouabain (but not the lipids) increased the affinity constant of Na,K-ATPase for K+, whereas the lipids (but not ouabain) increased that for Na+. The lipids also inhibited 86Rb uptake by intact human leukemia HL60 cells at potencies quite comparable to those seen for inhibition of purified protein kinase C or Na,K-ATPase. It is suggested that Na,K-ATPase (sodium pump) might represent a hitherto unrecognized site of action for the lipid analogues, and that the antineoplastic effects of the agents might be due to, in part, inhibition of both protein kinase C and Na,K-ATPase and perhaps other membrane-associated enzymes.

Biological Transport, Active↗

PubMed1990

Effects of antineoplastic phospholipids on parameters of cell differentiation in U937 cells.

The proliferation of the human promonocytic leukemia cell line U937 is inhibited by several ether lipids, ether lipid analogues and by phorbol esters. An early effect of this retardation of cell growth is the induction of a basic chromosomal protein, histone H1(0). Northern blot analysis of H1(0) mRNA levels reveals an increase of the mRNA concentration within a few hours after addition of hexadecylphosphocholine and 1-O-octadecyl-2-O-methyl-rac-glycero-3-phosphocholine. This early effect on the synthesis of a subtype of H1 proteins precedes the expression of several parameters of the monocytic differentiation of U937 cells.

Antigens, Surface↗

PubMed1990

New cytostatics--more activity and less toxicity.

Explore the source record for details and available documents.

Adenocarcinoma↗

PubMed1990

Hexadecylphosphocholine: a new and selective antitumor drug.

Interdisciplinary cooperation between basic and clinical research has resulted in the discovery and development of alkylphosphocholines, a new class of substances for the treatment of breast cancer. In contrast to most antitumor substances, the alkylphosphocholines do not attack the cell nucleus, but the cell membrane. This report presents a systematic study which, for the first time, provides a correlation between their chemical structure, antitumor efficacy and selectivity. Through an understanding of the metabolism of tumor growth inhibiting (ether)-lysolecithins, the minimal structural requirements for the antineoplastic efficacy of these substances have been obtained. This knowledge was used to identify molecular structures which are more effective and less toxic for the organism. The active principle derived from a study of (ether)-lysolecithins active as antitumor agents represents a new class of compounds: the alkylphosphocholines. As reported here, hexadecylphosphocholine is the most promising candidate of this group of compounds. It has an extremely selective action against chemically induced, autochthonous rat mammary carcinomas. No loss of activity was observed when comparing oral and intravenous administration. Particularly striking (and favorable for long-term therapy) is the fact that immunosuppression and hematotoxicity were not found at drug concentrations which lead to complete tumor remissions. Results obtained from animal experiments have been confirmed by preliminary clinical investigations.

Animals↗

PubMed1990

Hexadecylphosphocholine in the topical treatment of skin metastases in breast cancer patients.

Widespread local recurrence of breast cancer, untreatable by surgery or radiation therapy, can present a serious therapeutic problem predominantly in patients refractory to systemic therapy. In a phase I trial hexadecylphosphocholine, a new agent with high membrane affinity and antineoplastic activity was applied topically to affected skin areas of breast cancer patients. The results provide evidence that hexadecylphosphocholine may be an active agent in the topical treatment of skin metastases.

Administration, Topical↗