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WO1999006040A9 - Procede pour traiter les maladies a l'aide d'un tocotrienol, et d'un acide alpha-lipoique ou des derives ou un ester de ces derniers - Google Patents

Procede pour traiter les maladies a l'aide d'un tocotrienol, et d'un acide alpha-lipoique ou des derives ou un ester de ces derniers

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Publication number
WO1999006040A9
WO1999006040A9 PCT/US1998/016207 US9816207W WO9906040A9 WO 1999006040 A9 WO1999006040 A9 WO 1999006040A9 US 9816207 W US9816207 W US 9816207W WO 9906040 A9 WO9906040 A9 WO 9906040A9
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WO
WIPO (PCT)
Prior art keywords
composition
tocotrienol
disease
lipoate
alpha
Prior art date
Application number
PCT/US1998/016207
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English (en)
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WO1999006040A1 (fr
Inventor
Christopher J Berry
John L Foley
Lester Packer
Original Assignee
Christopher J Berry
John L Foley
Lester Packer
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Christopher J Berry, John L Foley, Lester Packer filed Critical Christopher J Berry
Priority to AU87680/98A priority Critical patent/AU8768098A/en
Publication of WO1999006040A1 publication Critical patent/WO1999006040A1/fr
Publication of WO1999006040A9 publication Critical patent/WO1999006040A9/fr

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Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/38—Heterocyclic compounds having sulfur as a ring hetero atom
    • A61K31/385—Heterocyclic compounds having sulfur as a ring hetero atom having two or more sulfur atoms in the same ring

Definitions

  • the present invention relates to a method of treating diseases, more specifically using a combination of a tocotrienol and alpha-lipoic acid or using a tocotrienyl lipoate or using derivatives thereof.
  • the present invention further relates, in particular to a method of reducing the occurrence of the excess incidence of cardiovascular disease and specific types of cancer in tobacco smokers.
  • 5,204,373 discloses the use of carba-tocotrienols as having superior metabolic stability over tocotrienols; United States Patent No. 4,603,142 discloses the use of a specific species of tocotrienol to lower cholesterol; United States Patent No. 5,334,61 2 discloses the treatment of retroviral diseases, such as HIV, using alpha-lipoic acid; United States Patent No. 5,084,481 discloses the treatment of pain and inflammatory disorders, including diabetic neuropathy and arteriosclerosis, using dihydrolipoic acid.
  • NF-kappa B and AP-1 nuclear transcription factors
  • SUBS 1 1 1 U 1 h SHEET (RULE 26)
  • AP-1 regulation in turn is modulated by the free radical status of a cell which can activate translocation of NF-kappa B from the cell cytosol to the cell nucleus where it can bind to DNA and facilitate transcription of gene products involved in disease states.
  • Antioxidants by affecting the free radical status of the cell, can regulate the activation of NF-kappa B and AP-1 .
  • a progressive deterioration in the antioxidant network characterizes the aging process. As a result of this deterioration, intracellular free radical levels rise. This in turn changes the regulation of the transcription of NF-kappa B and AP-1 related genes. More specifically, there is an age-associated alteration in nuclear factor kappa-B and AP-1 signal transduction pathways. See, e.g., Helenius et al., Biochem.
  • the age-associated decline is caused by altered regulation of the inhibitor l-kappa B, the redox-mediated activation step.
  • Trebilcock et al. Gerontology 42(3) : 1 37-146 ( 1996).
  • a functional antioxidant network assessed in terms of glutathione levels is required for NF-kappa B activation.
  • Ginn-Pease et al. Biochem. Biophys. Res. Commun. 226(3):695-702 (Sept. 24, 1 996).
  • Glutathione levels decline with age and disease.
  • TNF-alpha The transcription of gamma-glutamylcysteine synthetase is up-regulated by TNF-alpha. This up-regulation is attenuated by preincubation with high physiological levels of glucose. Thus, high glucose levels impair the regulation of glutathione. See, e.g., Morales et al., J. Biol. Chem. 272(48):30371 -9 (Nov. 28, 1997). Glutathione peroxidase overexpression in turn down-regulates thresholds of NF-kappa B activation by altering the isoform composition of l-kappa B. See, e.g., Suyang et al., Mol. Cell.
  • SCC Squamous carcinoma
  • PAHs polynuclear aromatic hydrocarbons
  • nitrates cause the cigarette to burn hotter, decreasing tar yields, but at the expense of increased generation of nitrogenous compounds, which are the gaseous carcinogens implicated in the rise of adenocarcinomas.
  • cigarette companies have reduced the tar yields which they are obliged to disclose but introduced a new hidden killer which apparently is responsible for a twenty-fold increase in the rate of adenocarcinomas.
  • PAHs the main carcinogen in tobacco tar
  • cytochrome P450-related compounds break down the PAHs into a form which can be solubilized by the enzyme glutathione S-transferase. Glutathione
  • Glutathione is the major antioxidant system in the body, and is synthesized in cells by enzymes from three dietary amino acids: glutamate, cysteine and gl ⁇ cine. Glutathione-related enzymes are encoded by a superfamily of genes. See, e.g., Lafuente et al., Cancer Lett. 68 (1 ):49-54 (Jan. 1 5, 1 993). These genes include GST M 1 and GST T1 . Almost half of the population inherits less than optimal glutathione-related genes. The GST M1 gene is deleted in 50%of the population and the GST T1 gene in 16%. Hayes et al., Crit. Rev. Biochem. Mol. Biol. 20 ⁇ 6):445-600 (1995). Thus, as aging occurs, or as the functionality of the antioxidant network is impaired by infection, carcinogens or metabolic stresses, or inherited suboptimal genetic polymorphisms, the function of the transcription factors NF-kappa B and
  • AP-1 is attenuated. See, e.g., Pinkus et al., J. Biol. Chem. 271 (23):13422-9 (June 7, 1 996). It has been suggested that these genetic abnormalities are major determinants of susceptibility to degenerative diseases such as cancers, for example tobacco-related lung cancers, and many studies have found elevated risks. Further, as the normal production of cytokines is disturbed, there is disregulation of the hypothalamic/pituitary/adrenal (HPA) axis, resulting in hypertension, diabetes, arterial sclerosis and heart failure. Inherited variations in the genes which regulate antioxidant enzyme transcription exacerbate these conditions. Subjects inheriting polymorphisms of the cytochrome P450 and glutathione transferase gene families are at elevated risk of degenerative disease.
  • HPA hypothalamic/pituitary/adrenal
  • Anttila et al. Cancer Res. 55(1 5):3305-9 (Aug. 1 , 1995); Nakajima et al., Carcinogenesis 1 6(4) :707-1 1 (Apr. 1 995); Kihara et al., Carcinogenesis 1 5(2):415-8 (Feb. 1 994); Kawajiri et al.. Princess Takamatsu Symp. 21 ( ): 55-61 (1990).
  • Cigarette smokers have elevated cortisol levels (see, e.g., Pomerleau et al., Pharmacol. Biochem. Behav. 36(1 ):21 1 -3 (May 1990)), and whole blood from smokers has been found to have a 38% higher TNF production after lipopolysaccharide stimulation. Tappia et al., Clin. Sci. (Colch.) 88(4):485-9 (April 1995).
  • SUBSTITUTE SHEET (RULE 25) the hormones which regulate glucose metabolism (insulin), and those that counter-regulate glucose metabolism (glucocorticoids). Epinephrine and norepinephrine mediate short-term adjustments, whereas cortisol influences longer term cyclic responses. The apparent insulin resistance and hypertension which characterize cigarette smokers. Syndrome X, adult onset diabetes and Cushing's Syndrome, share a common antecedent in disturbances to the HPA axis. Insulin resistance can be considered as disregulation of and/or an increased tissue sensitivity to glucocorticoids. Thioredoxin, a redox-regulating cellular cofactor that modulates cellular glucocorticoid responsiveness, influences tissue sensitivity to glucocorticoids.
  • the HPA axis is a complex open system, modulated by counter-regulatory hormones, by inhibitory feedback loops and by cytokines. Cytokines modulate glucocorticoid hormones and are modulated by them. Since cellular production of cytokines is controlled by transcription factors, principally NF-kappa B, and NF-kappa
  • B activation is controlled by cellular redox status, antioxidants modulate the HPA axis.
  • glucocorticoids The interaction between glucocorticoids and insulin is mediated through thresholds of NF-kappa B expression. NF-kappa B and the glucocorticoid receptor physically interact, resulting in the repression of NF-kappa B transactivation. Glucocorticoids inhibit cytokine gene transcription through receptor cross-coupling.
  • the activated glucocorticoid receptor can bind to and inactivate AP-1 and NF-kappa B.
  • Glucocorticoids strongly downregulate TNF-alpha. Pretreatment of cells with glucocorticoids modulates NF-kappa B activation, TNF-alpha and serum glucose. Administration of glucocorticoids alters hormonal and cytokine response to lipopolysaccharide in humans. Elevated TNF and cortisol levels have been found to exaggerate S-TNF-R responses for up to 144 hours after infusion, and in some tissue glucocorticoids upregulate the transcription of l-kappa B isoforms which bind to NF-kappa B in the cytosol and modulate NF-kappa B mediated gene transcription. Glucocorticoids thus are the link between insulin resistance and hypertension.
  • the HPA system as a whole, is in dynamic semi-stable equilibrium and pivots around set points analogous to thermostats. At birth, these set points are predetermined by the environment of the womb, when the fetus is exposed to the maternal regulatory system. Consequently, it has been demonstrated that low birth weight children have higher cortisol levels as adults, much higher incidence of adult- onset diabetes, and hypertension, stroke and heart disease as adults. Subsequently, contemporary stresses such as nocturnal hypoglycemia, disrupted sleep patterns, psychological stress, cigarette smoke, environmental toxins or aging, can disrupt the setpoints and influence HPA function by affecting NF-kappa B activation thresholds.
  • Excess cortisol has profound effects on the metabolism of glucose and protein. It increases gluconeogenesis by increasing glycogenolysis in muscle to provide lactate as a gluconeogenic precursor. It increases lipoiysis in the periphery to provide glycerol as a gluconeogenic precursor. Since cortisol has anti-insulin effects, it decreases the uptake of glucose into muscle, adipose, lymphoid and fibroblast tissues. Excess cortisol also breaks down muscle proteins to provide gluconeogenic precursors to the liver. Disregulation of these effects lead to hyperglycemia and a characteristic pattern of central obesity.
  • HPA axis The programming of the HPA axis has been shown to influence body weight, blood pressure body fat disposition patterns, insulin resistance and depression.
  • Diabetics have significantly greater 24-hour urinary-free cortisol output which increases with duration of diabetes and degree of diabetic complications.
  • Diabetic neuropathy is associated with specific and persistent inreases in HPA axis activity. Cortisol levels are elevated in symptomatic poiyneuorpathy.
  • SUBSTITUTE SHEET (RULE 25) Cytokines whose expression is controlled by NF-kappa B, particularly TNF- alpha, the cytokine implicated in insulin resistance, modulate the HPA axis and certain of the metabolic consequences of cortisol elevation are modulated by redox sensitive steps.
  • Glucocorticoids also have mineralocorticoid activities. Excess glucocorticoids may spill over onto aldosterone receptors to increase renal Na + absorption and increase renal K + and H + excretion, retaining water in the ECF, and elevating blood pressure. Also, glucocorticoids synergize catecholamines, increasing stroke volume and cardiac output which can also cause hypertension.
  • Cortisol levels rise in response to nicotine in smokers and are permanently elevated by cortisol-secreting tumors in Cushing's Syndrome, while in adult onset diabetes the diurnal cycle of cortisol secretion is disturbed and cortisol responses to TNF are exaggerated. Inappropriately elevated cortisol levels affect renin/angiotensin receptors inducing sodium retention and hypertension, counter-regulate insulin and attenuate insulin-mediated uptake of glucose by GLUT-4 receptors in muscle tissue.
  • Cigarette smoke condensate activates NF-kappa B, directly compounding the disregulation. See Shen et al., Am.J.Physiol. 270(5 Pt.2):H1624-33 (May 1996).
  • cortisol which normally responds to low blood glucose levels in order to protect the brain from hypoglycemia, is instead elevated by a false nicotine-mediated signal and insulin down-regulated when blood glucose levels are still high, resulting in the area under the curve (AUC) of blood glucose rising and futher imparing the transcription of protective antioxidant enzymes.
  • SUBST ⁇ SHEET (RULE 25 interactions activate NF-kappa B, as does glycated LDL, and activated NF-kappa B is found in arte ⁇ osclerotic lesions.
  • AGE-apolipoprotein B and serum AGE levels in cigarette smokers have been found to be significantly higher than those in non-smokers. See Cerami et al., "Tobacco Smoke is a Source of Toxic Reactive Glycation Products," The Picower
  • Cigarette smoke is a source of reactive AGE compounds. Id. It is therefore not supp ⁇ sing that diabetics who smoke have 2-4 times the incidence of cardiovascular disease than non-smoking diabetics.
  • the HPA is an open, adaptive system whose components interact in complex, self-regulating redundant patterns. The effects of isolated components can be paradoxical and shifted in time. For example, severely elevated cortisol levels can produce depression or euphoria, and normal responses to stimuli can be inverted or attenuated in diabetics and hypertensives or after prolonged oxidative stress.
  • the cellular processes which control the gene transcription of cytokines depend on the integrity of the antioxidant signaling network In many degenerative conditions, including the tobacco-related diseases, it is the attenuation of the antioxidant signaling network by increasing reactive oxygen species' stress loads that trigger disease onset. Redox sensitive pathways modulate the HPA axis directly, by controlling the transcription of cytokine messengers, and indirectly by ameliorating the consequences of HPA dysfunction by reducing glycation damage. Thus, strengthening and reinforcing this network can delay the onset of degenerative diseases.
  • SUBSTITUTE SHEET (RULE 25) which function particularly well in regulating activation of NF-kappa B and which can be non-toxically administered to a human or an animal.
  • a method of reducing the occurrence of cancers in human patients who are especially susceptible to such occurrence is also a need for a method of reducing the occurrence of lung cancer in human patients who smoke tobacco products and who are predisposed to lung cancer by inherited polymorphisms in cytochrome P450 1 A1 and glutathione S-transferase M, such as those described, e.g., in Lafuente et al., Cancer Lett. 68(1 ):49-54 (Jan. 15, 1993);
  • a disease selected from the group consisting of cancer, arterial sclerosis, abnormal platelet aggregation, hypertension, congestive lung disease, diabetes, viral infection, septic shock, graft versus host disease, cystic fibrosis, autoimmune disease, neurodegenerative disease, asthma, gout, kidney failure, Parkinson's disease, multiple sclerosis, and Alzheimer's disease.
  • the methods comprise the step of administering to a human or an animal, preferably a mammal, in need of such treatment a composition including an ingredient selected from the group consisting of a tocotrienyl lipoate, a combination of a tocotrienol and alpha- lipoic acid, derivatives thereof, and combinations thereof in an amount effective to reduce or prevent a symptom of such disease.
  • a method of regulating cellular apoptosis comprises the step of administering to a human or an animal a composition including an ingredient selected from the group consisting of a tocotrienyl lipoate, a combination of a tocotrienol and
  • SUBSTITUTE SHEET (RULE 25) alpha-lipoic acid, derivatives thereof, and combinations thereof in an amount effective to regulate cellular apoptosis.
  • the method comprise the step of administering to a human or an animal, preferably a mammal, in need of such treatment a composition including an ingredient selected from the group consisting of a tocotrienyl lipoate, a combination of a tocotrienol and alpha-lipoic acid, derivatives thereof, and combinations thereof, thereby regulating NF-kappa B.
  • a composition including an ingredient selected from the group consisting of a tocotrienyl lipoate, a combination of a tocotrienol and alpha-lipoic acid, derivatives thereof, and combinations thereof, thereby regulating NF-kappa B.
  • the composition is administered in an amount effective to reduce transcription of a gene product controlled by NF-kappa B by between about 5% and about 99%.
  • the gene product is selected from the group consisting of a cytokine, a cytokine receptor, a cell adhesion molecule, a viral protein, a growth factor, a growth factor receptor, an immunoreceptor, a transcription factor, an oncogene and nitric acid synthase.
  • a method of regulating cellular NO- metabolism comprising the step of administering to a human or an animal a composition including an ingredient selected from the group consisting of a gamma-tocotrienol, a gamma-tocotrienyl lipoate, a combination of a gamma-tocotrienol with alpha-lipoic acid, derivatives thereof, such as other isomers of vitamin E that are "free" at the ortho position on the phenolic moiety, and combinations thereof.
  • the method of regulating cellular NO x metabolism comprises regulating nitric oxide synthase activity.
  • the method of regulating cellular NO x metabolism comprises decreasing the formation of peroxynitrites.
  • compositions including an ingredient selected from the group consisting of a tocotrienyl lipoate, a combination of a tocotrienol and alpha-lipoic acid, derivatives thereof, and combinations thereof, and a pharmaceutically acceptable carrier.
  • a pharmaceutically acceptable carrier Preferably the composition is formulated in an amount from greater than about 1000 mg to about 2500 mg.
  • the foregoing derivatives are selected from the group consisting of linaloyl lipoate, farnesyl lipoate, geranyl lipoate, nerol lipoate, gamma-hydroxybutyrate ester of alpha lipoic acid, gamma-hydroxybutyrate ether of alpha lipoic acid, cysteine ester of tocopherol, cysteine ester of tocotrienol, succinate ester of tocotrienol, and succinate ether of tocotrienol.
  • the derivatives are preferably administered in an amount from about 25 mg to about 1000 mg.
  • a method of reducing the occurrence of a disease selected from the group consisting of adenocarcinoma and squamous cell carcinoma in a human who smokes tobacco products and is predisposed to lung cancer by at least one inherited polymorphism selected from the group consisting of a cytochrome P450 1 A1 polymorphism and a glutathione S-transferase M 1 polymorphism which comprises the step of administering to the human a composition including an effective detoxifying amount of an ingredient selected from the group consisting of a tocotrienyl lipoate, a tocotrienol in combination with alpha-lipoic acid, derivatives thereof, and combinations thereof.
  • SUBS I ITU Ik SHEET (RULE 25) step of administering to the human a composition including an effective amount of a component that promotes the intracellular conversion of cystine to cysteine.
  • the composition further includes an effective amount of a components that reduces the intracellular formation of at least one nitrosamine.
  • tocotrienyl lipoate or a combination of a tocotrienol and alpha-lipoic acid act synergistically as compared to a tocotrienol alone or alpha- lipoic acid alone in reducing or blocking inflammatory responses and in regulating the activity of transcription factors sensitive to a cell redox state, such as the DNA transcription factor NF-kappa B.
  • NF-kappa B is part of a family of Rel transcription factors that share common characteristics. They consist of hetero- or homodimeric proteins in association with an inhibitory protein family, l-kappa B. It is believed that one mechanism by which NF-kappa B is activated is by phosphorylation and dissociation of l-kappa B from Rel protein dimers, followed by l-kappa B's proteolytic degradation, which allows the dimeric DNA-binding protein NF-kappa B to enter the nucleus and regulate gene transcription.
  • NF-kappa B A host of genes have been shown to be modulated by NF-kappa B, including genes for cytokines and growth factors, immunoreceptors, adhesion molecules, acute-phase proteins, transcription factors and regulators, NO-synthase, and viral genes.
  • genes for cytokines and growth factors, immunoreceptors, adhesion molecules, acute-phase proteins, transcription factors and regulators, NO-synthase, and viral genes A general description of the NF-kappa B transcriptional regulation system can be found in Bauerele and Baltimore, Cell, Volume 87, 13-20 (1996), incorporated herein by reference.
  • ROS reactive oxygen species
  • RNS reactive nitrogen species
  • acquired immunodeficiency syndrome results from infection with a human immunodeficiency virus (HIV-1 or HIV-2), which eventually destroys a specific subset (CD4 + ) of helper T-lymphocytes, so that the patient ultimately yields to opportunistic infection and certain neoplasms.
  • HIV-1 proviral DNA contains two binding sites for NF-kappa B, which activate transcription by binding to the sequence ⁇ '-GGGACTTTCC-S' in the kappa enhancer where it interacts with the transcription apparatus.
  • the virus usurps normal cellular machinery in order to transcribe its own DNA.
  • LDL Low density lipoprotein
  • Oxidants appear to be involved in two steps of the atherosclerotic process.
  • Low density lipoprotein (“LDL”) particles enter the artery wall and some remain.
  • LDL becomes mildly oxidized, possibly by ROS or RNS released by vascular cells.
  • the mildly oxidized LDL contains a component or components, believed to be oxidized phospholipid, which causes the endothelium to secrete molecules that result in the recruitment of monocytes and their differentiation to macrophages. Further oxidation of the LDL results in its uptake by macrophages, their conversion to foam cells, and deposition in the vascular wall as part of fatty-
  • NF-kappa B appears to be a central transcription factor in the expression of a number of the genes induced by the early oxidation step in atherosclerosis, including serum amyloid A and macrophage colony-stimulating factor.
  • C57BL/6J mice and C3H/HeJ mice which are susceptible and not susceptible to formation of aortic fatty streaks, respectively, are fed an atherogenic diet, only the C57BL/6J mice show activation of NF-kappa B.
  • VCAM-1 induction of expression of VCAM-1 in response to a variety of signals is mediated by an NF-kappa B-like DNA-binding protein and is blocked by a variety of antioxidants, including pyrrolidine dithiocarbamate and NAC.
  • Accelerated arterial sclerosis in diabetes and cigarette smokers may be due to the presence of larger amounts of AGE, a greater degree of interaction of these with the receptor for AGE (AGE-RAGE interaction), activation of NF-kappa B, expression of VCAM-1 , and priming of the diabetic vascuiature for enhanced interaction with circulating monocytes.
  • these diseases By interfering with the activation of NF-kappa B, these diseases, and other diseases which are believed to be caused at least in part by the effect of reactive oxygen species on NF-kappa B (for example by NF-kappa B activation of gene transcription) can be treated.
  • the following diseases in addition to the above-listed are believed to be so caused: cancer, cancer metastasis, abnormal platelet
  • NF-kappa B regulation of NF-kappa B with the compositions of the present invention can cause or inhibit apoptosis, depending upon the particular cell type and the redox state of the cell, thereby regulating cellular apoptosis.
  • an animal is susceptible to disease states caused by NF-kappa B activation, such animal may be similarly treated.
  • a disease may be considered treated by the inventive methods if a symptom of the disease is improved to the extent that an improvement in the symptom is measurable by a standard test or perceptible to a human with the disease.
  • a standard test or perceptible to a human with the disease For example, an increase in CD4 + T-cell levels as measured by standard assay, in a patient with AIDS. See, A Manual of Laboratory and Diagnostic Tests, Fifth Ed. Francis Fishcbach (1 996), incorporated herein by reference.
  • a disease may be considered prevented by the inventive methods if a disregulation or imbalance leading to the condition is corrected.
  • tocotrienols and alpha-lipoic acid are the synergistic effect of tocotrienols and alpha-lipoic acid which provides enhanced lipophilic mobility and intermembrane transferability to the tocotrienols.
  • tocotrienols are distributed evenly throughout the biomembrane whereas alpha tocopherol is clumped in relatively immobile clusters. Accordingly, tocotrienols are more readily available to more cellular compartments to interact with free radicals in those compartments.
  • compositions of the present invention can be preferably orally administered or administered by another route to a human or an animal in an amount to provide a concentration in vivo sufficient to modulate NF-kappa B activity.
  • the regulation of cellular nitrogen monoxide ("NO") and nitrogen dioxide (“N0 2 ”) metabolism (“NO.") is important with respect to carcinogenesis, particularly carcinogenesis caused by exposure to environmental air pollutants.
  • N0 2 can form peroxynit ⁇ tes which are potent free radicals and can damage cells by inducing lipid peroxidation and protein oxidation. Peroxynitntes can also nitrosate amines; nitrosated amines can cause DNA mutations, which can lead to carcinogenesis. N0 restricted radicals can also activate NF-kappa B which in turn can activate transcription of inducible nitrogen monoxide synthase.
  • Inducible nitrogen monoxide synthase is long lasting, Ca 2 + independent and glucocorticoid sensitive Nitrogen monoxide synthase generates NO leading to a positive feedback cycle of inducible nitrogen monoxide synthase production.
  • constitutive nitrogen monoxide synthase activity is short lasting, having one sixth to one tenth the activity of inducible nitrogen monoxide synthase, is Ca 2+ dependent and is hormone insensitive.
  • gamma-tocot ⁇ enols due to the absence of a methyl group at the C-5 position can form a C-nitroso aromatic compound which thereby decreases the production of detrimental peroxynitntes, thereby regulating "NO/ metabolism
  • This effect can be augmented by administering or estenfying alpha-lipoic acid with the gamma-tocotrienol, due to the ability of the alpha-lipoic to regenerate oxidized tocotrienol radicals to tocotrienols.
  • Gamma-tocot ⁇ enols are believed to preferentially operate by the same mechanisms as set forth above Therefore administration of gamma-tocot ⁇ enols can reduce free radical induced DNA mutage ⁇ esi ⁇ and resulting carcinogenesis.
  • alpha lipoic acid and tocotrienols in a compound or mixture according to the present invention is effective in treating cancers, in particular lung cancer, both therapeutically and prophylactically That is, it has been discovered that such administration is effective in protecting against both the AC and SCC forms of lung cancer associated with cigarette smoking.
  • Such compositions, in particular gamma-tocotrienol lipoate are especially effective in maintaining glutathione-related detoxification mechanisms in cigarette smokers who have inherited sub-optimal combinations of the cytochrome P450 and glutathione gene superfamily. Further, the gamma-isomer of tocotrienol appears to offer additional protection against nitrosamine-induced adenocarcinomas.
  • S-transferase to join glutathione to the toxicant, boosting or restoring intracellular glutathione levels indirectly through alpha-lipoic administration, may help protect those smokers with an inherited attenuation of the glutathione system which detoxifies PAHs, thereby reducing their vulnerability to squamous carcinomas. Since the epidemiological pattern indicates that squamous carcinomas occur after the detoxification system has been exhausted by long-term exposure to cigarette smoke, it follows that administration of alpha-lipoic may have a very significant role in boosting glutathione-related detoxification mechanisms thereby postponing or preventing the incidence of squamous carcinomas.
  • Tocotrienols have been reported to have various anti-cancer effects including the modulation of glutathione S-transferase levels in animal cancer models.
  • gamma-tocotrienol is 40 to 60 times more readily recycled in membranes than alpha-tocopherol, and the gamma-isomers of tocopherols and tocotrienols appear to function to detoxify nitrogen oxides present in cigarette smoke by protecting against the formation of carcinogenic nitrosamines.
  • alpha-lipoic acid alpha-lipoic acid
  • SUBSTI TUTE SHEET (RULE 25) appears to recycle tocotrienols. The effects of tocotrienols and alpha-lipoic have been shown to be synergized when co-administered.
  • each of these compounds synergizes the other within the domain of its respective protective action.
  • the addition of tocotrienols to alpha-lipoic acid potentiates the modulation by alpha-lipoic acid of the glutathione-related pathways and reinforces the effectiveness of cytochrome P450, while the addition of alpha-lipoic acid to tocotrienols facilitates the systemic recycling of tocotrienols, thereby facilitating detoxification of nitrogen metabolites.
  • the compensatory modulation of gene expression achieved by this pairing is unique and previously unrecognized.
  • an effective amount of the inventive composition for use in preventing the occurrence of lung cancers as discussed herein is about 5 mg to about
  • the molar ratio of alpha-lipoic acid to tocotrienol is about 1 : 100 to about 2:1 .
  • the R-form of alpha-lipoic acid is particularly potent.
  • inventive compositions described herein are administered in divided doses 30-45 minutes prior to meals.
  • other modes of administration of the inventive compositions will also be readily apparent to those skilled in the art.
  • inventive compositions can be incorporated into a functional food for consumption by a patient.
  • inventive compositions can also include additional antioxidants and/or nutriceuticals, such as vitamin C, vitamin B 6 and biotin, which can potentiate the therapeutic efficacy of the compositions.
  • additional antioxidants and/or nutriceuticals such as vitamin C, vitamin B 6 and biotin, which can potentiate the therapeutic efficacy of the compositions.
  • Useful dosages typically will be in the range from about 1 to 10 RDA's (recommended daily allowance) per day.
  • Anti-inflammatory activity of tocotrienyl lipoates or tocotrienols and alpha-lipoic acid materials was conducted using the skin cell line 2 ® ZK1301 .
  • the cytotoxicity of the test materials is estimated by the MTT50 of each material ⁇ i.e., the concentration required for 50% reduction in the cell viability of treated tissues relative to the untreated controls.
  • Tissue Model skin 2 ® Model ZK 1 301 Test Material: 1 ) T3 as 5.2% rice tocols in 84% PEG 600, 1 1 % non- tocol rice lipids.
  • Rice tocols have a distribution of 47% gamma- tocotrienol, 23% aplha-tocopherol, 22% gamma tocopherol, 6% delta-tocopherol, 2% alpha-tocotrienol and 1 % delta-tocotrienol, other tocotrienol-like compounds may be present.
  • T3 and alpha-lipoic acid as 4.9% rice tocols and 4.7% alpha lipoic acid in 80%PEG 600, 1 1 % non-tocol rice lipids.
  • PMA phorbol myristal acetate
  • the calibration and operation of all equipment is checked and documented prior to beginning the study.
  • the assay procedures for 1 -[4,5 Dimethylthiazol-2-yl]-3,5- diphenylformazan (MTT) and PGE 2 are as follows. The tissues are removed from the agarose shipping tray and placed in MILLICELL ® plates containing serum-free assay media with 10 ng/ml PMA under each MILLICELL ® . An applicator pad dosed with
  • 25 ⁇ l of PMA is then placed on the epidermal side of the tissues.
  • the tissues are then incubated overnight in a 37°C, 5% C0 2 , -- 90% humidity incubator.
  • the media containing the PMA is aspirated after 24 hours and replaced with fresh serum free assay media without PMA.
  • the tocotrienol, or tocotrienol and alpha lipoic acid are dispensed directly on the epidermal side of the tissues.
  • Three tissues are dosed for each test material and control. Once all tissues have been dosed, the 0 time control media samples are collected and frozen at -20°C. Fresh serum-free assay media without PMA are added and the plates are placed in a 37°C, 5% C0 2 , ⁇ 90% humidity incubator for the exposure time indicated above. The media are collected at each time point and frozen at -20°C for later analysis for PGE 2 . After the last time point,
  • the percent untreated control value for all of the dilutions of each test material is calculated as follows:
  • the MTT optical density values for the untreated control tissues are assumed to represent 100% viability.
  • PGE 2 release is measured with a commercially available immunoassay. Concentrations of PGE 2 in the test samples are determined from a standard curve generated with each run. Data is expressed as total pg released per tissue.
  • Prostaglandin E 2 (PGE 2 ) release is an indicator of a response to PMA challenge involving membrane perturbation, events that activate phospholipase A 2 , protein kinase C (“PKC”) activation, and the expression of gene products regulated by NF- Kappa B activation.
  • PGE 2 assay set forth below (Table 1 ) show a synergistic effect of tocotrienols and alpha-lipoic acid in suppressing synthesis of
  • MTT CELL VIABILITY AFTER 24 HRS INCUBATION mean MTT o.d. 540 nm.
  • MTT is a substrate for mitochondrial succinate dehydrogenase and is converted to an insoluble formazan by the activity of this enzyme. The amount of formazan produced is proportional to the number of viable cells in the tissue.
  • Tocotrienyl lipoates may be synthesized using dicyclohexylcarbodiimide (DCC) and 4-dimethylaminopyridine (DMAP) to facilitate the esterification of alpha-lipoic acid to the tocotrienol or tocopherol in a manner similar to that used by V.
  • DCC dicyclohexylcarbodiimide
  • DMAP 4-dimethylaminopyridine
  • Azais-Braesco for the synthesis of Vitamin A esters of fatty acids in JAOCS 69:1 272-1273 (1992), incorporated herein by reference.
  • ALA Alpha-lipoic Acid
  • DCC Dicyclohexylcarbodiimide
  • Solution A was swirled and then sonicated in a bath type sonicator to dissolve all compounds fully.
  • Solution A was then continuously purged with N 2 until solution B was added. Due to this purging the temperature of solution A was about 10-15 degrees C.
  • Solution B was prepared with 0.9-1 .1 gm of DCC (approximately 4.8 mmol) and
  • reaction solution was filtered through a fast sped paper filter, then a medium speed paper filter, and the filter was washed with 5-10 ml of MeCI 2 to recover product.
  • the filtrate was washed sequentially with water, then acid, 0.25
  • Solution A was swirled then sonicated in a bath type sonicator to dissolve all compounds fully. Solution A was then continuously purged with N 2 until solution B
  • Solution B was prepared with 2.06 gm of DCC in 20 ml of MeCI 2 .
  • Solution B was swirled to dissolve DCC fully, then 10.5 ml of B (1 .03 gm, approximately 4.8 mmol) was added to Solution A while swirling.
  • 10.5 ml of solution B had been added, the flask was again purged with N 2 , capped with an air tight stopper and placed in the dark overnight. This flask was swirled several more times at irregular time intervals to ensure good mixing. Shortly after all of solution B was added, the reaction solution became cloudy with a precipitate that was believed to be urea.
  • the reaction solution was filtered through a fast speed filter, then a medium speed paper filter, and the filter was washed with 5-10 ml of MeCI 2 to recover product.
  • the filtrate was washed sequentially with water, then acid, 0.25 N HCI, then a basic solution of approximately 0.9N Na 2 C0 3 , and then water in a beaker using a spin bar to provide agitation to remove unwanted compounds.
  • the washed MeCI 2 was dried over Na 2 SOores to remove water, the solvent was removed (MeCI 2 evaporated) on a hot water bath with a stream of N 2 . Prior to being fully dry, a precipitate was observed. This was removed by filtration in the same manner as above and solvent evaporation continued.
  • the yellow colored oil product was then analyzed by HPLC.
  • Solution A was prepared with:
  • Solution A was swirled then sonicated in a bath type sonicator to dissolve all compounds fully. Solution A was then continuously purged with N 2 until solution B was added. Due to this purging, the temperature of solution A was about 10 - 1 5 degrees C.
  • Solution B was prepared with 2.06 gm of DCC in 20 ml of MeCI 2 .
  • Solution B was swirled to dissolve DCC fully, then 9.0 ml of solution B (0.881 gm, 3.8 mmol) was added to solution A while swirling.
  • 9 ml of solution B had been added, the flask was again purged with N 2 , capped with an air tight stopper and placed in the dark overnight. This flask was swirled several more times at irregular time intervals to ensure good mixing. Shortly after all of solution B was added, the reaction solution became cloudy with a precipitate that was believed to be urea.
  • reaction solution was filtered through a fast speed paper filter, then a medium speed paper filter, and the filter was washed with 5-10 ml of MeCI 2 to recover product.
  • the filtrate was washed sequentially with water, then acid, 0.25
  • HEWLETT PACKARD 1046A FLUORESCENCE DETECTOR Set at 295/340 nm
  • Vydac C18 Column: 218 TP54 250X 4.6 mm, 5 um particle size, Seperations Group MOBILE PHASE: Acetronitrile/Methanol/Ethyl Acetate & Acetic Acid 1 : 1 95:4.5:0.25/0.25 @ 1 .6 ml/min.
  • Standards ALPHA T-1 STD Sigma
  • ALPHA T-1 ACETATE STD Sigma
  • Synthesis 1 and 2 used rice tocols (RT3). It was found that the reaction products as set forth above contained tocotrienyl lipoates and tocopheryi lipoates. Confirmation that the desired product had been produced can be obtained from the data collected from the chromatograms and spectra of the parent tocotrienols and tocopherols and the reaction products. The change in retention times, and shift in spectral character indicate that a compound other than tocotrienols and tocopherols were present.
  • the spectral similarity of the tocotrienyl and tocopheryi lipoates to their corresponding acetates (and succinates) in the 260-290 nm range is indicative of ester formation at the phenolic OH group of tocotrienols and tocopherols.
  • the lipoates have characteristic absorbance in the 300-400 nm range which are similar to alpha-lipoic acid. This absorbance is only slightly effected by esterification. There was virtually no trace of the original tocotrienols and tocopherols in the reaction products indicating near quantitative synthesis of the desired product. Confirmation
  • SUBST ⁇ UTE SHEET (RULE 26) of the ester nature of the product was obtained by the regeneration of the parent tocotrienols and tocopherols after saponification, with the concurrent disappearance of the presumed tocotrienyl lipoates and tocopheryi lipoates.
  • the lipoates have characteristics in the 300-400 nm range, similar to alpha-lipoic acid. There was virtually no trace of the original tocopherols in the reaction products indicating near quantitative synthesis of the desired product.
  • Individual isomers e.g., alpha, beta, gamma, delta, epsiion, of tocotrienols and tocpherols or combinations of isomers of tocotrienols and tocpherols may be used in combination with alpha-lipoic acid or esterified with alpha-lipoic acid.
  • Gamma- tocotrienol may preferably be used.
  • the R or S form of alpha-lipoic acid may be used. The R form is preferred.
  • isoprenoids having a phenolic or alcohol group can be synthesized by the method set forth above.
  • the following compounds which it is believed can perform similar functions to tocotrienyl lipoates or tocotrienol and alpha-lipoic acid, can also be used and can be synthesized by the method set forth above.
  • These include terpene alcohol esters or ethers of alpha lipoic acid.
  • these esters are: tocotrienyl succinate, linaloyl lipoate made from linanol, Molecule No. 5520 in the 12th Ed. of the Merck Index; farnesyl lipoate, made from farnesol, Molecule No. 3978 in the 1 2th Ed.
  • geranyl lipoate made from geranol, Molecule No. 441 1 in the 1 2th Ed. of the Merck Index
  • nerol lipoate made from nerol, Molecule No. 6560 in the 1 2th Ed. of the Merck Index
  • suitable compounds include gamma-hydroxybutyrate ester or ether of alpha lipoic acid, cysteine ester of tocopherol and tocotrienol and the succinate ester or ether of tocotrienol.
  • compositions of the present invention can be administered orally, topically, parenterally, by suppository and by other standard routes of administration.
  • Tocotrienyl lipoate is preferentially absorbed topically as compared to tocotrienol.
  • Combinations of administration routes which yield the greatest bioavailability of a compound or combination of compound can be used. For example, administration of compound 1 by an oral route and administration of compound 2 by a suppository is preferred.
  • compositions of the present invention can be administered as an ingredient of along with a pharmaceutically acceptable carrier.
  • Pharmaceutically acceptable carriers as are known in the art can be used with the present invention.
  • compositions for oral administration include capsules, tablets, dispersible powders, granules, syrups, elixirs and suspensions. These compositions can contain one or more conventional adjuvants, such as sweetening agents, flavoring agents, coloring agents and preserving agents.
  • Tablets can contain the active ingredients in a mixture with conventional pharmaceutically acceptable excipients. These include inert carriers, such as calcium carbonate, sodium carbonate, lactose, and talc; granulating and disintegrating agents, such as starch and alginic acid; binding agents such as starch, gelatin acacia; and lubricating agents, such as magnesium stearate, stearic acid and talc. Tablets may be uncoated or coated by known ' techniques to delay disintegration and absorption in the gastrointestinal tract thereby providing a sustained action over a longer period of time.
  • inert carriers such as calcium carbonate, sodium carbonate, lactose, and talc
  • granulating and disintegrating agents such as starch and alginic acid
  • binding agents such as starch, gelatin acacia
  • lubricating agents such as magnesium stearate, stearic acid and talc. Tablets may be uncoated or coated by known ' techniques to delay disintegration and
  • Capsules may contain the active ingredients alone or an admixture with an inert solid carrier, such as calcium carbonate, calcium phosphate or kaolin.
  • an inert solid carrier such as calcium carbonate, calcium phosphate or kaolin.
  • SUBSTITUTE SHEET (RULE 25) suspensions, syrups and elixirs may contain the active ingredients in mixture with any of the conventional excipients utilized in the preparation of such compositions.
  • compositions of the present invention are administered in at least such amount that a symptom of a disease, as would be defined by standard medical practice, is improved to the extent that an improvement in the symptom is measurable by a standard test or perceptible to a human with the disease and not in an amount to cause harm to a human or animal.
  • compositions of the present invention can be administered in at least such amount as to regulate NF-kappa B activity by reducing transcription of a gene product controlled by NF-kappa B by between about 5% and about 99%, preferably between about 25% and about 99%, more preferably between about 50% and about 99%.
  • Reduction of gene transcription can be determined as set forth in Arenzana-Seisdedos, F. et al. Mol. Cell Biol. 1 5:2689-2696 (1 995) and Suzuki et al. Biochem. Biophys. Research Comm. 189: 1 709-1 5 (1992), each of which is incorporated herein by reference.
  • an effective dosage is from about 25 mg to about 2500 mg per day. Dosages may vary from 125 mg to 1250 mg or from 250 mg to about 500 mg per day.
  • compositions for use in treating smokers are provided.
  • compositions described above can be used for the treatment or prevention of lung cancers in tobacco smokers.
  • compositions also include the following additives in the amounts stated: Vitamin C 5-10 RD As

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Abstract

L'invention concerne un anti-oxydant ou une combinaison d'anti-oxydants qui permettent de moduler l'action de radicaux libres, et en particulier, qui permettent de réguler l'activation d'un facteur de transcription, tel que le NF-Kappa B et qui ne sont pas toxiques et peuvent être administrés à l'homme ou à l'animal. L'invention concerne plus particulièrement un procédé pour traiter des maladies, y compris pour réduire l'incidence des cancers du poumon chez les fumeurs. Ce procédé utilise un tocotriénol et un acide alpha-lipoïque ou un tocotriényl lipoate ou des dérivés de ces derniers.
PCT/US1998/016207 1997-08-04 1998-08-04 Procede pour traiter les maladies a l'aide d'un tocotrienol, et d'un acide alpha-lipoique ou des derives ou un ester de ces derniers WO1999006040A1 (fr)

Priority Applications (1)

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AU87680/98A AU8768098A (en) 1997-08-04 1998-08-04 Method of treating disease using a tocotrienol and alpha-lipoic acid r derivatives or an ester thereof

Applications Claiming Priority (2)

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US5543397P 1997-08-04 1997-08-04
US60/055,433 1997-08-04

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FR2786104B1 (fr) 1998-11-25 2002-12-27 Centre Nat Rech Scient Inhibiteurs de l'activation de nf-kb, et leurs utilisations pharmaceutiques
US6284786B1 (en) * 1999-02-16 2001-09-04 The Center For The Improvement Of Human Functioning, Int'l., Inc. Treatment of cancer using lipoic acid in combination with ascorbic acid
FR2791571B1 (fr) * 1999-04-02 2002-10-04 Sod Conseils Rech Applic Association inhibiteur(s) de no synthase et antioxydant(s) metabolique(s)
IT1312060B1 (it) * 1999-04-09 2002-04-04 Antibioticos Spa Uso dell'acido alfa lipoico nel trattamento antimetastatico.
WO2000078296A2 (fr) * 1999-06-17 2000-12-28 Basf Aktiengesellschaft Tococotrienol et/ou derives de tocotrienol pour le traitement therapeutique ou prophylactique de troubles provoques par le glutamate et/ou le calcium
DE19941217A1 (de) * 1999-08-30 2001-03-15 Asta Medica Ag Behandlung der Migräne durch Verabreichung von alpha-Liponsäure oder Derivaten derselben
JP2001342133A (ja) * 2000-03-31 2001-12-11 Eisai Co Ltd γ―トコトリエノール含有利尿剤
WO2001074354A1 (fr) * 2000-03-31 2001-10-11 Eisai Co., Ltd. DIURETIQUES CONTENANT DU η-TOCOTRIENOL
DE10027968A1 (de) * 2000-06-08 2001-12-13 Asta Medica Ag Mittel zur Therapie von Demenzen
DE20203244U1 (de) 2002-03-01 2002-05-23 Meins, Wolfgang, Prof. Dr., 22391 Hamburg Pharmazeutische Zusammensetzung zur Prävention der Alzheimerdemenz
US20040034093A1 (en) * 2002-05-30 2004-02-19 Hensley Kenneth L. Methods for enhancing motor performance and/or endurance
RU2325908C1 (ru) * 2006-09-20 2008-06-10 Государственное учреждение научно-исследовательский институт онкологии Томского Научного центра Сибирского отделения Российской академии медицинских наук (ГУ НИИ онкологии ТНЦ СО РАМН) Способ профилактики осложнений при комбинированном лечении больных раком легкого и желудка
DE102007038849A1 (de) 2007-08-16 2009-02-19 Adscil Gmbh Verwendung von R(+)-alpha-Liponsäure zur Behandlung der kryptogenen Neuropathie

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US5217992A (en) * 1989-10-04 1993-06-08 Bristol-Myers Squibb Company Tocotrienols in the treatment of hypercholesterolemia, hyperlipidemia and thromboembolic disorders
WO1993009777A1 (fr) * 1991-11-22 1993-05-27 Lipogenics, Inc. Composes de tocotrienols et analogues aux tocotrienols, et leurs procedes d'utilisation

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