Compositions which inhibit apoptosis, methods of purifying...

Drug – bio-affecting and body treating compositions – Antigen – epitope – or other immunospecific immunoeffector – Conjugate or complex

Reexamination Certificate

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C435S325000, C435S360000, C435S410000

Reexamination Certificate

active

06306398

ABSTRACT:

FIELD OF THE INVENTION
This invention relates to compositions of matter which are effective in inhibiting apoptotic cell death.
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 cell death may be initiated by a wide variety of signals, including hormones, serum growth factor deprivation, chemotherapeutic agents, ionizing radiation, and infection by human immunodeficiency 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; and 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.
All references cited herein both supra and infra are hereby incorporated herein by reference.
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.
Lysophosphatidic acid (LPA) is found in a variety of plant products as are a variety of phospholipids. LPA as been found to have a variety of physiological activities including mitogenesis, growth factor and as an anti-wrinkle agent. U.S. Pat. Nos. 4,263,286; 4,746,652; 5,326,690; and 5,340,568. LPA is reviewed in detail by Moolenaar (1994)
TICB
4:213-219; and Eichholtz et al. (1990)
Biochem. J.
291:677-680.
SUMMARY OF THE INVENTION
The present invention encompasses methods of obtaining compositions that inhibit apoptosis and the compositions obtained thereby. The compositions are termed phytogenic apoptosis inhibitors (PAIs). The invention encompasses physiologically acceptable compositions suitable for administering the PAIs in an amount sufficient to modulate apoptosis. The invention further encompasses methods of use of the PAIs.


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Funk et al., “Effect of soy products on methotrexate toxicity in rats”J. Nutr.(1991)

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