Compositions which inhibit apoptosis, methods of making the comp

Drug – bio-affecting and body treating compositions – Preparations characterized by special physical form – Liposomes

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424401, 514 75, 514 78, 514844, 514863, 514880, 4352402, 4352403, A61K 9127

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060045795

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BRIEF SUMMARY
FIELD OF INVENTION

This invention relates to compositions of matter which are effective in inhibiting apoptotic cell death. More specifically, it relates to compositions of phospholipids (PA, PI, LPA, LPI and LPC) the mixture of which exhibits anti-apoptotic activity.


BACKGROUND OF THE INVENTION

Apoptosis is a normal physiologic process that leads to individual cell death. This process of programmed cell death is involved in a variety of normal and pathogenic biological events and can be induced by a number of unrelated stimuli. Changes in the biological regulation of apoptosis also occur during aging and are responsible for many of the conditions and diseases related to aging.
Recent studies of apoptosis have implied that a common metabolic pathway leading to apoptosis may be initiated by a wide variety of signals, including hormones, serum growth factor deprivation, chemotherapeutic agents, ionizing radiation, and infection by human imnmunodeficiency virus (HIV). Wyllie (1980) Nature 284:555-556; Kanter et al. (1984) Biochem. Biophys. Res. Commun. 118:392-399; Duke and Cohen (1986) Lymphokine Res. 5:289-299; Tomei et al. (1988) Biochem. Biophys. Res. Commun. 155:324-331; Kruman et al. (1991) J. Cell. Physiol. 148:267-273; Ameisen and Capron (1991) Immunol. Today 12:102-105; and Sheppard and Ascher (1992) J. AIDS 5:143-147. Agents that affect the biological control of apoptosis thus have therapeutic utility in numerous clinical indications.
Apoptotic cell death is characterized by cellular shrinkage. chromatin condensation, cytoplasmic blebbing, increased membrane permeability and internucleosomal DNA cleavage. Gerschenson et al. (1992) FASEB J. 6:2450-2455; and Cohen and Duke (1992) Ann. Rev. Immunol. 10:267-293.
A variety of food supplements containing, in part, partially processed plant extracts have been used to ameliorate the gastrointestinal disorders that often accompany chemotherapy, radiation and AIDS. The supplements generally contain carbohydrates, fat and plant protein hydrolysates. See, e.g., Tomei and Cope et al. in Apoptosis The Molecular Basis of Cell Death (1991) Cold Spring Harbor Laboratory Press.
Several proteinase inhibitors derived from plant extracts have anticarcinogenic activity. Troll et al. (1987) Adv. Cancer Res. 49:265-283. The Bowman-Birk inhibitors are the best described of these inhibitors. Birk (1985) Int. J. Pep. Pro. Res. 25:113-131. Bowman-Birk inhibitors are described as a family of disulfide bonded proteins with a molecular weight of about 8 kD which suppress cellular transformation. Chou et al. (1974) Proc. Natl. Acad. Sci. USA 71:1748-1752; Yavelow et al. (1985) Proc. Natl. Acad. Sci. USA 82:5395-5399; and Yavelow et al. (1983) Cancer Res. (Suppl.) 43:2454s-2459s. Crude soybean extracts containing Bowman-Birk inhibitors have been described. Kennedy et al. U.S. Pat. No. 4,793,996; PCT publication No. WO 94/20121; and Kennedy, A. R. (1994) Cancer Res. (Suppl) 54: 1999s-2005s. Bowman-Birk inhibitors have also been described immunologically. WO 90/03574; and U.S. Pat. Nos. 4,959,310; and 5,053,327. Bowman-Birk inhibitors have also been found to have activity in degranulation of macrophages. Japanese Patent No. 63051335.
Phospholipids are a class of amphipathic phosphorous-containing lipids which are essential constituents of biological membranes. Various phospholipid preparations have been used for cooking, drug delivery (liposomes), slow release delivery systems, carrier media for hydrophobic drugs, gene transfer and replacement therapy, sunscreens, emulsions, anti-foaming agents, replacement of damaged or absent pulmonary surfactants, detergents and membrane stabilization.
Phosphatidic acid (PA), phosphatidylinositol (PI), lysophosphatidic acid (LPA), lysophosphatidylinositol (LPI), and lysophosphatidylcholine (LPC) are found in a variety of plant and animal products. LPA is found in a variety of plant and animal products and has been reported to have a variety of physiological activities including mitogenesis, growth factor and to be an anmi-wrinkle agent. U.S. P

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