WO1999034675A1

WO1999034675A1 - Promoting mobilization and catabolism of lipids

Info

Publication number: WO1999034675A1
Application number: PCT/US1999/000423
Authority: WO
Inventors: William E. Shell; Mark E. Jarmel
Original assignee: Nutracorp Scientific, Inc.
Priority date: 1998-01-07
Filing date: 1999-01-07
Publication date: 1999-07-15
Also published as: AU2217399A

Abstract

Methods and compositions for mobilization and catabolism of lipids in a subject. A lipolytic agent is administered to mobilize lipids together with a substrate for an energy-consuming chemical-reaction in the body that activates the energy-consuming reaction to thereby promote the catabolysis of the mobilized lipids. Lipolytic agents include xanthines, indirect sympathomimetic amines and precursors for norepinephrine and for histamine. The substrates include amino acids that are metabolized to form ammonia and amino acids involved in either or both the glutamine-glutamate and urea reaction cycles, such as glutamine, glutamic acid, aspartic acid, glycine, histidine, arginine, citrulline, ornithine. Glucagon secretion is additionally promoted, to further effect lipolysis, through the administration of ornithine and arginine, which also function as substrates. Glucagon secretion is also promoted by administering limited quantities of mono- and di-saccharides.

Description

PROMOTING MOBILIZATION AND CATABOLISM OF LIPIDS

Technical Field

This invention relates generally to agents for releasing and burning fats (lipids) stored in the body in order to reduce percentage of body fat and decrease weight. There has been increasing attention to weight control since obesity is associated with an increased mortality rate, diabetes mellitus, hypertension, heart disease and stroke. Despite the desirability of reducing weight and the proliferation of products to aid in weight reduction, the occurrence of obesity continues to rise. Pharmaceutical agents used for weight reduction have known undesirable side effects. Additionally, appetite suppressing agents have had limited success in weight management since overweight individuals tend to eat when they are not hungry. Accordingly, there is a need for an agent that promotes weight loss by a means other than suppressing feelings of hunger. There is a need for an effective means of releasing fat from storage and stimulating burning of the released fat.

Background Art

Lipolysis is the release of fat from fatty tissue (adipose tissue) into the blood stream in the form of fatty acids. It is known that lipolysis can be induced in humans by injection of norepinephrine (Millet L, et al. Catecholamine effects on lipolysis and blood flow in human abdominal and femoral adipose tissue. J.Appl.Physiol. 1998;85:181-188). However, Applicants are not aware of any prior art suggesting that lipolysis can be induced in humans through oral administration of a norepinephrine precursor. Xanthines have been characterized as having the ability to stimulate lipolysis and to augment the lipolytic effect of norepinephrine. Histamine has also been identified in the literature as having a lipolytic effect when injected in test subjects. Amphetamines and related appetite-suppressing agents such as phentermine are known to produce weight loss in humans (Valle-Jones JC, et al. A comparative study of phentermine and diethylpropion in the treatment of obese patients in general practice. Pharmatherapeutica. 1983;3:300-304 and Wellman PJ. Overview of adrenergic anorectic agents. Am.J.Clin.Nutr. 1992;55:193S-198S). There is no disclosure in these references that these agents cause weight loss by promoting lipolysis, at least in the absence of a concomitant exercise regimen.

U.S. Patent No 5,229,390 to Moriyama et al. relates to a beverage or food product containing additives for mobilizing body fats so that the fats may be utilized more efficiently, particularly for consumption during exercise. In fact, simply stimulating the release of fats from adipose tissue, at absent such exercise, is unlikely to result in a reduction in body fat. Mobilized fats would circulate for a time in the bloodstream and would then be returned to storage as body fat.

U.S. Patent No 5,229,390 relates to a beverage or food product containing an additive for mobilizing body fats so that the fats may be utilized more efficiently, particularly for consumption during an exercise program. The additive includes certain amino acids, and particularly arginine, alanine, an leucine, that accelerate the release of glucagon and xanthines. Both glucagon and xanthines are lipolytic agents and are used in these products for mobilizing fats. These products are reported to increase metabolism of fatty acids when taken during an exercise program. However, it is not apparent that the additive would appreciably reduce body fat, absent the exercise program. In fact, simply stimulating the release of fats from adipose tissue is unlikely to result in a reduction in body fat. Mobilized fats would circulate for a time in the bloodstream and then be returned to storage as body fat.

It is known that substrate cycling reactions, also known as futile shuttles, exist in the body and cause a net consumption of energy (Sibson NR, et al. In vivo 13C NMR measurements of cerebral glutamine synthesis as evidence for glutamate-glutamine cycling. Proc.Natl.Acad.Sci.U.S.A. 1997;94:2699-2704). Applicants are not aware of any prior art suggesting how these substrate cycling reactions might be clinically activated or, if activated, how they can result in a reduction of percentage body fat and/ or weight loss. Simply activating energy consuming reactions would not be expected to reduce body fat, since circulating blood sugar would tend to be used for energy production rather than circulating fat.

In a mice study it was found that glutamine supplementation of a high fat diet reduced body weight and attenuated hyperglycemia. (Opara, EC et al, L-Glutamine supplementation of a high fat diet reduces body weight and attenuates hyperglycemia in C57BL/6J Mice J. Nurtr. 126:273- 279, 1996). The mechanism for these results was reported to be unclear since glutamine is an inhibitor of fatty acid oxidation. These observations would suggest that any weight loss caused by glutamine was due to mechanisms other than fatty acid oxidation.

Disclosure of the Invention

It has been discovered that stimulating release of fats from storage and simultaneously stimulating energy- consuming reactions, may significantly reduce weight and percentage of body fat. Concomitantly administering a lipolytic agent, an agent that releases fatty acids from adipose tissue, together with a substrate used by an energy-consuming reaction, surprisingly reduces percentage of body fat and decreases scale weight. By concomitant administration is meant administration of two or more components either simultaneously, the most desirable procedure, or within a reasonably short time of the other, desirably within 15 minutes or a shorter period.

Using this invention one may release fat from storage and stimulate fat burning. Thus, using this invention one may achieve a reduction in percentage of body fat and a decrease in body weight. This invention provides methods and compositions for releasing fat from storage and promoting fat burning for energy production. With this invention, one may achieve these effects using agents that are not considered either prescription or non-prescription drugs. Using this invention one may release fat from storage and stimulate fat burning using readily available, low cost, safe agents found in the ordinary diet. Further, with this invention one may achieve these effects at reduced dosage of the agents, thereby minimizing the possibility of side effects.

By activating mechanisms that consume energy in the form of ATP, a relative energy deficit is created. This results in a need for energy production. Fatty acids that have been released by the lipolytic agent are available in the bloodstream for energy production. Thus, by stimulating energy-consuming reactions simultaneously with stimulating release of fat from storage, a "siphon-like" effect occurs. This siphon-like effect further promotes release of fats from storage. Therefore, by stimulating release of fat from storage and simultaneously stimulating energy-consuming reactions, one may reduce percentage of body fat and decrease weight.

Energy-consuming reactions particularly suitable for this invention are the glutamine-glutamate cycle and /or the urea cycle. The glutamine-glutamate cycle and the urea cycle comprise a class of substrate cycling chemical reactions endogenous to the body that consume energy. The substrates that may be employed in this invention include any of the constituents or intermediates of the metabolic pathways of these cycles. Substrates used by other energy- consuming cycles that are endogenous to the body, may also be used as a substitute for, or together with, substrates for the glutamine-glutamate cycle and/ or the urea cycle.

Norepinephrine promotes release of fats stored in adipose tissue. In a feature of this invention, methods and compositions are provided for promoting release of stored fat by administering a norepinephrine precursor. Tyrosine and phenylalanine each are precursors for both norepinephrine and dopamine.

In another feature of the invention, histidine (the precursor for histamine), may be orally administered with a norepinephrine precursor. Administering histidine with a norepinephrine precursor preferentially promotes conversion of the precursor to norepinephrine instead of dopamine. Histamine promotes norepinephrine release and inhibits dopamine release. Additionally, histamine may augment release of fat from adipose tissues.

In a related feature, the precursors for norepinephrine may be orally administered simultaneously with one or more xanthines, and particularly caffeine, theobromine, and/ or theophylline, in order to increase activity of norepinephrine. Additionally the xanthine may augment mobilization of fat from adipose tissue.

In yet another feature, histidine and xanthines may be employed together with the norepinephrine precursors for a further enhanced effect. Histidine and xanthines operate through different mechanisms in enhancing norepinephrine release and are thus complementary.

In a further feature of this invention, a norepinephrine agonist or indirect sympathomimetic agent, particularly epinephrine, ephedrine, pseudoephedrine, isoproterenol HC1, or metaproterenol sulfate, may be administered, as the lipolytic agent. Such adrenergic agents may be administered simultaneously with a substrate used by the glutamine-glutamate cycle or urea cycle in order to increase fat burning.

In a further feature of this invention, a xanthine may be used as the fat-releasing agent. One may simultaneously administer a xanthine with a substrate used by an energy-consuming reaction in order to stimulate release and burning of stored fat. Theobromine has been found to be surprisingly useful. Theobromine may promote release of fat from storage without central nervous system stimulation, thereby avoiding undesired side effects.

Glutamine and glutamic acid comprise a class of substrates involved in the energy-consuming glutamine-glutamate cycle. In a feature of this invention, glutamine or glutamic acid may be administered as the substrate for an energy-consuming reaction. One may simultaneouly administer glutamine or glutamic acid along with a fat-releasing agent to stimulate release and burning of stored fat.

Glutamine, glycine, histidine and glutamic acid are metabolized to form ammonia. In another feature of this invention, glutamine, glycine, histidine or glutamic acid, may be administered to promote formation of ammonia within the body. The conversion of ammonia to urea consumes ATP, which creates a relative energy deficit. The relative energy deficit caused by processing ammonia promotes burning of fats that have been released from storage by the lipolytic agent.

Further, glutamine is converted into glutamate in the cytoplasm and glutamate is transported into the mitochondria by the malate- aspartate shuttle. Aspartate is transported out of the mitochondria into the cytoplasm in exchange for glutamate. Using this invention one may create an excess of glutamine in the cytoplasm that is available to interact with aspartate to form asparagine. Conversion of aspartate to asparagine consumes ATP, thereby further promoting creation of a relative energy deficit. In yet another feature, arginine may be simultaneously administered with glutamine and/ or glutamic acid. Arginine promotes transport of glutamine from the bloodstream into metabolically active cells of the various body systems. Thus, arginine potentiates the effects of glutamine used by this invention. Arginine additionally acts as a substrate for the urea cycle, which consumes ATP. Processing arginine through the urea cycle thus consumes energy, which stimulates burning of fatty acids released by this invention for energy production.

Amino acids such as arginine, citrulline, ornithine, aspartic acid, glutamine, and glutamic acid comprise a class of substrates for the urea cycle and may be administered simultaneously with a lipolytic agent in accordance with this invention. These substrates for the urea cycle may be administered simultaneously with a lipolytic agent to promote catabolism of mobilized lipids. Ornithine, arginine and glutamine are preferred due to their synergistic action in promoting the use of mobilized fats for fuel.

It is another feature of this invention to stimulate secretion of glucagon as glucagon will further promote the mobilization of fat. Ornithine, is known to stimulate secretion of glucagon and may be utilized in this invention for this purpose. Thus, it may perform the dual role of being a substrate for the urea cycle and for additionally promoting secretion of glucagon. Ornithine stimulates secretion of glucagon without appreciably promoting secretion of insulin. Ornithine may thus be utilized in a dual role, without dosage restriction, both as a urea cycle substrate in this invention and as promoter of lipolysis. For this invention, excessive release of insulin is undesirable since insulin is anti-lipolytic and inhibits glucagon secretion. For this reason, the amount of carbohydrate and /or insulin secretogogues (substances that promote insulin secretion) is desirably limited in some embodiments of this invention, as will be discussed.

Arginine also stimulates glucagon. Thus, arginine can also be used advantageously in the dual role of an energy cycle substrate and additionally as a promoter of glucagon secretion for lipolysis. However, arginine is an insulin. Thus, higher doses of arginine are appropriately limited to avoid excessive release of insulin that would inhibit glucagon secretion. Desirably, for this purpose arginine is administered in this invention in a limited dosage, desirably at least 25 mg., but less than about 1 gram and preferably less than about 600 mg.

In a related feature of this invention a carbohydrate, and particularly a mono- and /or disaccharide, is concomitantly administered to the subject. The secretion of glucagon can be stimulated by a relative drop in blood sugar. It has been found that a relatively minor release of insulin will stimulate a blood sugar decrease. Thus, in this feature of the invention a limited amount of carbohydrate is concomitantly administered to cause release of a relative small amount of insulin. This, in turn, causes drop in blood sugar that will stimulate glucagon secretion to act as a lipolytic agent. But the carbohydrate is administered in an amount too low to cause the excessive insulin release that would result in an antiUpolytic effect and suppression of glucagon release. Desirably, the dosage amount of such carbohydrates is at least about 250 mg. but desirably less than about 6 grams and preferably less than about 3 grams, as dextrose equivalent.

Formulations of this invention using the foregoing ingredients for promoting glucagon may produce an additional benefit by affecting eating-related satiation. Surprisingly and unexpectedly subjects ingesting such formulations reported experiencing early onset of satiation during a meal and /or prolonged satiety after completion of a meal. Additionally, favorable effect on appetite may result from the use of the pharmaceutical adrenergic lipolytic agents used in this invention. These can be significant additional benefits in the practice of this invention.

Excessive release of insulin is thus avoided in this invention by restricting concomitant intake by the subject of carbohydrates and insulin secretagogue amino acids that would otherwise result in an excessive release of insulin. Accordingly, the compositions of this invention are desirably administered to the subject on an empty stomach and concomitant intake by the subject of carbohydrates and insulin secretagogue amino acids is restricted, desirably to a combined dosage amount of less than about 6 grams.

To insure effectiveness of the compositions of this invention, the concomitant intake by he subject of foodstuffs is desirably restricted. Such intake may adversely affect absorption by the subject of the active agent. In particular, to avoid competitive absorption, the concomitant intake by the subject of amino acids, other than the energy cycle substrates and lipolytic agents administered, is desirably restricted to an amount less than about 50% by weight of the substrate amino acids administered. Preferably, concomitant intake of such other amino acids is completely excluded. Similarly, to prevent interference with absorption of the lipolytic agents and substrates, concomitant intake of fiber is desirably limited to about 10% by weight of the substrate amino acids administered and is preferably excluded completely.

Brief Description of the Drawing

FIGURE. 1 is a box diagram illustrating the flow of energy- consuming reactions believed to take place in one embodiment of this invention, leading to the consumption of mobilized fat.

Best Mode of Carrying Out The Invention

A lipolytic agent, and particularly a precursor and /or agonist for norepinephrine, may be administered concomitantly with a substrate of an energy-consuming reaction in the body and particularly a substrate used by the glutamine-glutamate cycle and /or the urea cycle, to enhance release and burning of stored body fat. It has been discovered that such co-administration promotes a reduction of percentage of body fat and a decrease in scale weight.

While Applicants do not wish to be bound by any particular theory, FIGURE 1 illustrates their current theoretical understanding of how the present invention may operate in one preferred embodiment. This embodiment utilizes two energy-consuming cycles. In the glutamine-glutamate cycle glutamine is converted to glutamate, releasing ammonia, and then glutamate is converted back to glutamine (box 1). In the urea cycle ammonia is converted to urea (box 6). In this embodiment glutamine, the substrate for the glutamine-glutamate cycle, is ingested (box 5) causing production of glutamate and ammonia (NH₄ ⁺) and then conversion of the glutamate back to glutamine (box 1). Both of these reactions consume energy in the form of ATP. The ammonia is then converted into urea in the urea cycle (box 6) consuming an even larger amount of energy, in the form of ATP. Each turning of the urea cycle consumes four molecules of ATP. Concomitantly, a norepinephine precursor is ingested that promotes production of norepinephrine (box 2), which releases fat from storage (box 3). The released fat is burned to produce ATP (box 4) which compensates for the deficit of ATP caused by activation of the energy consuming cycles. The ATP deficit thus caused by these cycles is believed to promote the fat burning to produce ATP. Thus, released fat is used to fuel energy-consuming reactions without the production of external work.

Pharmaceutical agents produce their effects by either activating or inhibiting mechanisms that are already present in the body. They therefore, attempt to emulate actions normally produced by the body's intrinsic homeostatic mechanisms. All pharmaceuticals that are foreign to the body have associated undesirable side effects since synthetic drugs imperfectly interact with the body's regulatory mechanisms. Side effects are further promoted by the nonselective distribution of drugs throughout the bloodstream. These pharmaceuticals thus may affect each organ and body system in unknown and potentially undesirable ways. This invention has the advantage of permitting the administration of naturally occurring agents, namely neurotransmitter precursors and substrates that are normally used by the body's intrinsic homeostatic mechanisms. Administering precursors normally used by the body to synthesize neurotransmitters along with substrates that are endogenous to the body desirably decreases the occurrence of side effects. Thus, this invention advantageously emulates effects produced by synthetic pharmaceutical agents on various mechanisms regulated by neurotransmitters, with reduced risk of side effects.

In the preferred modality of this invention for promoting lipolysis, the lipolytic properties of norepinephrine are employed. Thus, norepinephrine precursors and agonists and utilized in this invention and they function by promoting the activity of norepinephrine.

The neurotransmitter norepinephrine is active in the central nervous system and peripherally. In this invention precursors for norepinephrine, and particularly tyrosine and phenylalanine, are administered for the purpose of promoting the production and release of norepinephrine to optimize lipolysis. Advantageously, for this purpose the norepinephrine precursors may be administered concomitantly with one or more xanthines. Xanthines are not only lipolytic and are useful as an lipolytic agent in this invention, but they have also been found by Applicants to enhance the production and release of norepinephrine, thus synergistically enhancing its effect.

The norepinephrine agonists employed in this invention for lipolysis include the indirect-acting sympathomimetic amines which are known to function as appetite suppressants. They include drugs with both indirect-acting and direct-acting norepinephrine-releasing components. This class of agents includes epinephrine, ephedrine, pseudoephedrine, isoproterenol, metaproterenol, phentermine, phenylpropanola ine, amphetamine, pseudephedrine, norpseudoephedrine, diethylpropion, benzphetamine, phendimetrazine, phenmetrazine, chlorphentermine, and aminorex and their physiologically acceptable salts, hydrates, acid adducts, mineral chelates and their other active derivitives.

Alternatively in this invention, histidine the precursor for the neurotransmitter histamine may be administered as a lipolytic agent in this invention. As in the case of norepinephrine, administration of the precursor enhances the production of histamine, which is also lipolytic. Desirably histidine may be administered together with one or more of the other precursors above, in place of or together with a xanthine. It has been discovered that histidine is also capable of effectively increasing synthesis and release of norepinephrine, mediated through the increased production of histamine. Histidine and xanthines are desirably used together to increase synthesis and release of norepinephrine to achieve yet further enhanced effect.

Glutamine and glutamic acid are substrates used by the glutamine-glutamate cycle and form ammonia, which is processed by the urea cycle. Additionally, glycine and histidine are metabolized in the body to form ammonia. It has been found that these ammonia- generating substrates will enhance catabolism of lipids released from adipose tissue when administered together with a lipolytic agent and preferably a precursor and/ or agonist for norepinephrine.

The amino acids arginine, citrulline, ornithine, aspartic acid, glutamine and glutamic acid comprise a class of substrates for the urea cycle. One or more of these amino acids may be administered concomitantly with a lipolytic agent in accordance with this invention to enhance catabolism of lipids mobilized by the lipolytic agent.

Glutamine is converted into glutamate in the cytoplasm and glutamate is transported into the mitochondria by the malate-aspartate shuttle. Aspartate is transported out of the mitochondria into the cytoplasm in exchange for glutamate. This invention creates an excess of glutamine in the cytoplasm that is available to interact with aspartate to form asparagine, which further consumes ATP.

Ornithine and arginine are also preferred due to their synergistic action in promoting the catabolism of lipids. When concomitantly administered with glutamine and /or glutamic acid, arginine promotes transport of glutamine from the bloodstream into metabolically active cells of the various body systems, thus potentiating the effects of glutamine. Ornithine additionally promotes the release of glucagon which promotes lipolysis, as will be discussed.

The neurotransmitter precursors and the substrates of this invention may be employed in this invention in pure form, e.g. exogenous material synthesized or derived from animal or vegetable protein, particularly purified extracts isolated from the amino acid residues in enzyme hydrolyzed proteins. It is to be understood that these agents may also be in the form of their physiologically acceptable derivitives, including their salts, hydrates, acid adducts (e.g. hydrochloric acid) and mineral chelates (e.g. salts bound to the precursor by chelation bonding).

Xanthines constitute a class of non-selective adenosine antagonists and they include theobromine, caffeine, and theophylline. They are capable of promoting release of the neurotransmitters norepinephrine and histamine. Xanthines administered in accordance with this invention potentiate neurotransmitter synthesis and release for each of norepinephrine and histamine. Combining one or more xanthines, with one or more neurotransmitter precursors allows the desired effects to be achieved with reduced, safe, doses of the individual agents.

The xanthines may be used in the form of their free compounds or as their salts, adducts or other derivatives, for example citrated caffeine, theophylline ethylenediamine, theophylline sodium acetate, sodium glycinate, the choline salt, the theophylline derivatives theophylline megumine and dyphylline, theobromine calcium salicylate, sodium acetate or sodium salicylate.

A particularly suitable form of xanthines for use in this invention are those that are derived from natural sources. Cocoa provides a unique combination of the xanthines theobromine and caffeine in a form that is normally easily ingested and tolerated by the subject. Cocoa powder was originally included in preliminary formulations with neurotransmitter precursors to improve flavor and because its mood enhancing effects have appealed to people for centuries. An unexpected result was that the cocoa powder significantly potentiated the effects of the neurotransmitter precursors. This potentiating effect was determined by us to be produced by the naturally occurring xanthines present in cocoa powder.

Infusions of caffeine from coffee beans and of caffeine and theophylline from tea leaves may be employed as a natural source of these xanthines, either in liquid form as coffee and tea, or in dried extract form, alone or, more conveniently, in composition with the neurotransmitter precursor. Caffeinated soft drinks, chocolate, guarana, ephedra, mate' and other food or herb sources may be employed.

Xanthines may be employed in this invention in dosage ranges appropriate to promote release of neurotransmitters and to avoid undesired side effects. Theobromine may be administered in a dosage of preferably from about 1 mg. to about 2 grams or higher. Caffeine may be administered preferably in a dose of from about 1 mg. to about 200 mg. or higher if tolerated by the subject. Theophylline may be administered in a dose preferably of from about 1 to about 200 mg or higher if tolerated by the subject. Cocoa may be administered in a dose preferably of about 1 mg. to about 10 grams or higher for an appropriate dose of xanthines, with a preferred dose being about 500 to about 800 milligrams. Somewhat higher doses of these xanthines may be employed with some subjects without undue discomfort.

In general for the lipolytic agents of this invention, the desired dosage is an amount which will increase the mobilization of lipids in the subject. The appropriate dosage for lipolytic effect will depend on the particular class of agent. The desired dosage range for norepinephrine precursors, and particularly tyrosine or phenylalanine, is from about 50 to about 2,000 milligrams, with a typical dose of about 500 to 1,000 milligrams. However, doses up to 2,000 milligrams and even higher, e.g. up to 3 grams may be administered without undue risk of side effects.

The desired single dose range for a sympathomimetic agent utilized as the lipolytic agent in this invention desirably is typical for that agent where it is used as an appetite suppressant in the treatment of obesity. Appropriate dosage ranges for these agents are found in the literature, including in U.S. Patent 5,019,594 to Wurtman et al. For example, an appropriate single dose for phenylpropanolamine is from 5 to 25 mg., for amphetamine from 1.25 to 10 mg. and ephedrine 5 to 50 mg. More generally, as a class, these agents are administered in dosages as low as 3 mg, and as high as 50 mg. Where a sympathomimetic agent is utilized in this invention along with another lipolytic agent, such as a norepinephrine precursor and/ or a xanthine, the dosage amount of sympathomimetic agent may be reduced, proportionally.

The dosage of histidine, either for use with norepinephrine precursors and/ or with sympathomimetic agents in order to additionally enhance norepinephrine activity, is an amount sufficient to enhance synthesis and release of histamine in the body. Histidine may be administered in accordance with this invention to enhance thereby lipolysis, for which norepinephrine precursors, and /or sympathomimetic agents are simultaneously administered. The desired single dose range for this purpose for histidine is typically from about 1 to about 500 milligrams and may be up to about 1,000 milligrams, with a typical dose preferably being about 30 to about 200 milligrams. The desired single dose range for glycine administered in accordance with this invention is preferably from about 1 to about 500 milligrams and may be up to about 1,000 milligrams, with a typical dose preferably being about 30 to about 200 milligrams.

In general for the energy cycle substrates of this invention, the desired dosage for the purposes of this invention is an amount which will enhance the catabolization of lipids in the subject. The desired single dose range for each of the energy cycle substrates, other than arginine, i.e. citrulline, ornithine, glycine, aspartic acid glutamic acid and glutamine is from about 100 to 2,000 milligrams, with a typical dose of between 500 and 1,000 milligrams. The desired single dose range for arginine administered as a urea cycle substrate in accordance with this invention is from about 25 to 1,000 milligrams, with a typical dose of 25 to 600 milligrams. As arginine is a strong insulin secretagogue, higher doses may lead to excessive insulin secretion that suppression lipolysis, as will be discussed.

It is further desirable in this invention to stimulate secretion of glucagon as glucagon will further promote lipolysis. This is accomplished by two modalities. The first is employment as a energy- consuming cycle substrate of an amino acid that additionally promotes secretion of glucagon. Ornithine additionally promotes secretion of glucagon. Thus ornithine may be utilized in a dual role both as a urea cycle substrate in this invention and as promoter of lipolysis. As ornithine does not appreciably promote secretion of insulin, it may be employed without restriction as dosage. Thus, the same dosage range indicated above for use of ornithine as a urea cycle substrate in this invention may apply for this dual use.

Similarly, arginine, another substrate of the urea cycle, may be used in the dual role, as an energy cycle substrate and additionally as a promoter of glucagon for lipolysis. Arginine does promote insulin secretion and at limited dosage will release a relatively small amount of insulin that will be sufficient to cause a relative blood sugar drop. The blood sugar drop will then stimulate secretion of glucagon. For this purpose a dose of from about 25 mg. up to about 1 gram and preferably less than about 600 mg. may be desirable. At this level of administration, a relatively small amount of insulin is released by arginine causing a relative blood sugar drop that will stimulate glucagon secretion to promote lipolysis. By thusly limiting the arginine dosage, excessive insulin release is avoided, which would, instead, inhibit glucagon secretion.

In the second modality for promoting glucagon, a carbohydrate, and particularly a mono- and /or disaccharide, is concomitantly administered to the subject in an appropriate limited dose. This dose will cause a relative blood sugar drop. The blood sugar drop will, in turn, cause release of a relatively small amount of insulin that will stimulate glucagon secretion. The appropriate carbohydrate dose is insufficient to cause insulin release in an amount that would result in an antiUpolytic effect and suppression of glucagon release. Dextrose is the preferred carbohydrate, because it is a relatively strong insulin secretagogue and for volume considerations, a lesser quantity may be required. However, other mono- or di-saccharides may be employed in amounts that provide a corresponding insulin secretory effect. Desirably, the dosage amount of such carbohydrates is at least about 250 mg. but desirably less than about 6 grams and preferably less than about 3 grams, as dextrose equivalent. By dextrose equivalent is meant an amount of any particular mono- or di-saccharide to be employed that will produce an equivalent secretory effect as would the amount of dextrose specified.

The neurotransmitter precursors and other active agents of this invention may be administered orally separately, or for assurance of appropriate proportions and dosages as well as for convenience, they may be administered together in the same composition. The dosage forms for administration separately or in the same composition may be any of the conventional forms, including carbonated beverages, capsules, caplets, chewable wafers, tablets, liquid suspensions, powders and the Uke. Xanthine dosages may take the form of chocolate preparations, cocoa drinks, and cola drinks containing caffeine, either separate or incorporating the precursors and other desired ingredients.

The compositions in the form of powders or Uquids may be packaged in multiple dosage quantities with instructions to the user to extract therefrom for ingestion appropriate individual dosage amounts, e.g. a teaspoonful. However, the compositions are desirably prepared in discreet units, e.g. prepackaged beverages, capsules, wafers, etc., which each contain the appropriate dosage amounts of neurotransmitter precursors with xanthines and/ or histidine, for a single dose as discussed above. The compositions may include the usual carriers, in fiUers, excipients, flavorings and adjuvants in addition to neurotransmitter precursors, and other active agents.

It is preferable in carrying out this invention to administer the dosages when the subject has an empty stomach, typically at least an hour after the subject has eaten. Administering the compositions on an empty stomach is preferred in order to avoid undesirably slow uptake of precursors into the bloodstream. Uptake of administered neurotransmitter precursors would be inhibited by competition for absorption by other amino acids from the ingested food. It is a particular advantage of this invention that exercise is not a requirement for its effectiveness. However, further benefit may be seen by having the subject engage in exercise concomitantly with adminstration of the compositions of this invention.

The effects of the formulations of this invention normally should be sufficiently potent that their effects can be evaluated after two or three weeks. Objective measures of body fat may advantageously be used to assess the effectiveness of this invention. Examples of such objective measures are bioelectrical impedance, dual energy X-ray absorptiometry, underwater weighing, abdominal girth measurement, deuterium dilution in body fluids and determination of total body potassium.

The following examples iUustrate various embodiments of the invention.

Example 1

A formulation was prepared in the proportions of about 80 parts of cocoa with about 50 parts glutamine, about 50 parts tyrosine, about 10 parts glycine, about 5 parts histidine, about 40 parts arginine, and about 300 parts dextrose. Gelatin capsules are filled with the powder blend so that 6 gelatin capsules contain cocoa about 800 mg, glutamine about 500 mg, tyrosine about 500 mg, glycine about 100 mg, arginine about 400 mg, histidine about 50 mg, and sucrose about 3,000 mg. This formulation is preferably administered_when the subject has an empty stomach, preferably about 30-60 minutes after eating. Nine adult subjects ingested this formulation once a day for seven and a half weeks. At the end of that time, average loss of scale weight was 13 (+/- 6) pounds and average loss of body fat was 12 (+/- 5) pounds. The average reduction in percent body fat was 4 (+/- 2) percent.

Norepinephrine is known to decrease heart rate. Change in heart rate was used to assess the formulation's abiUty to increase norepinephrine levels sufficiently to produce significant physiologic alterations. Heart rate was measured using an ECG event recorder (King of Hearts) and a Pace Art receiving center. Following ingestion of the formulation, heart rate decreased by 16.7% at 30 minutes as compared to baseline, where placebo did not significantly change heart rate (Table 1).

Table 1

Thus, the formulation in Example 1 was able to reduce heart rate consistent with increased norepinephrine production.

Example 2

A formulation was prepared in the proportions of about 100 parts of cocoa with about 50 parts glutamine, about 50 parts tyrosine, about 10 parts glycine, about 5 parts histidine, about 5 parts arginine, and about 400 parts sucrose. Gelatin capsules are filled with the powder blend so that 6 gelatin capsules contain cocoa about 1,000 mg, glutamine about 500 mg, tyrosine about 500 mg, glycine about 100 mg, arginine about 50 mg, histidine about 50 mg, and sucrose about 4,000 mg. This formulation is preferably administered as in Example 1. The formulation was administered to an adult male who experienced early onset of satiation and greatly reduced meal size.

Example 3

A formulation was prepared in the proportions of about 80 parts of cocoa with about 50 parts glutamine, about 50 parts tyrosine, about 10 parts glycine, about 2.55 parts histidine, about 40 parts arginine, and about 400 parts sucrose. Gelatin capsules are filled with the powder blend so that 6 gelatin capsules contain cocoa about 800 mg, glutamine about 500 mg, tyrosine about 500 mg, glycine about 100 mg, arginine about 400 mg, histidine about 25 mg, and sucrose about 2,500 mg. This formulation is preferably administered as in Example 1. The formulation was administered to eight adults who each experienced early onset of satiation, prolonged post-meal satiety, and greatly reduced meal size (Table 2).

* p<0.05. The data were obtained by a 5 point questionnaire completed foUowing ingestion of a dose of Formula-3. Comparison of Formula-3 to placebo was performed by unpaired t-test.

Example 4

A formulation was prepared using aspartic acid and ornithine as substrates for the energy-consuming urea cycle. The formulation was prepared in the proportions of about 70 parts of cocoa with about 50 parts ornithine, about 50 parts tyrosine, about 50 parts histidine, about 25 parts aspartic acid, and about 25 parts dexcrose. Gelatin capsules are filled with the powder blend so that 4 gelatin capsules contain cocoa about 700 mg, ornithine about 500 mg, tyrosine about 500 mg, aspartic acid about 250 mg, histidine about 500 mg, and dextrose about 250 mg. This formulation is preferably administered as in Example 1. The formulation was administered to an adult male and an adult female who both experienced increased duration of post-meal satiation. Example 5

This example illustrates the use of formulations of this invention such as those described in Examples 1 through 4 in combination with a sympathomimetic agent as an additional lipolytic agent along with a norepinephrine precursor. Such combined administration allows reduced doses of the sympathomimetic agent to effectively promote loss of body fat with reduced risk of side effects. Phentermine about 7.5 mg may be advantageously administered with each of the formulation of Examples 1 through 4 to effectively promote appetite suppression and augment catabolism of lipids stored in adipose tissue. This dosage of phentermine is reduced by about 50% or more from the amount typically used in the treatment of obesity. Alternatively, other sympathomimetic agents such as ephedrine may be substituted for phentermine in formulations such as in Examples 1 through 4 and administered in amounts similarly reduced by about 50% or more compared to the doses of these agents typically used in the treatment of obesity. Thus, the potential risk of side effects associated with sympathomimetic pharmaceuticals is greatly reduced by this invention. An adult male with a stable baseline weight experienced a 4 pound weight loss in 10 days of ingesting the formulation in Example 1 with the addition of about 7.5 mg of phentermine.

Example 6

This example illustrates the use of a sympathomimetic agent as the primary lipolytic agent in formulations of this invention. A powder blend is prepared in the proportions of about 0.75 parts phentermine, about 60 parts of cocoa with about 50 parts glutamine, about 10 parts ornithine, about 5 parts histidine, about 5 parts arginine, and about 80 parts dextrose. Gelatin capsules are filled with the powder blend so that 4 gelatin capsules contain phentermine about 7.5 mg, cocoa about 600 mg, glutamine about 500 mg, ornithine about 100 mg, arginine about 50 mg, histidine about 50 mg, and dextrose about 800 mg. This formulation is preferably administered as in Example 1. This dosage of phentermine is reduced by about 50% or more from the amount typically used in the treatment of obesity. Alternatively, other sympathomimetic agents such as ephedrine may be substituted for phentermine and administered in amounts similarly reduced by about 50% or more compared to the doses of these agents typically used in the treatment of obesity. In addition to the enhanced catabolism of lipids stored in adipose tissue of such formulations, the sympathomimetic agent promotes appetite suppression.

Example 7

A formulation may be prepared that is designed to promote secretion of glucagon. A powder blend is prepared in the proportions of about 60 parts of cocoa with about 50 parts glutamine, about 40 parts tyrosine, about 10 parts ornithine, about 5 parts histidine, about 5 parts arginine, and about 80 parts dextrose. Gelatin capsules are filled with the powder blend so that 4 gelatin capsules contain cocoa about 600 mg, glutamine about 500 mg, tyrosine about 400 mg, ornithine about 100 mg, arginine about 50 mg, histidine about 50 mg, and dextrose about 800 mg. This formulation is preferably administered as in Example 1.

The formulation was given to an adult male who experienced an increase in fat mobilization as measured by triglyceride levels in plasma (Table 3). The subject also experienced an associated increase in oxygen consumption, reflected in an increase in nitric oxide production. As fatty acids are metabolized, oxygen consumption and the production of resulting oxygen radicals increases. Nitric oxide has anti-oxidant effects and is released in order to scavenge oxygen radicals produced by fatty acid oxidation. Thus, ingestion of the formulation induced increased oxygen consumption as reflected by an increase in nitric oxide production.

Table 3

Baseline 30min 60min

Formula 7 02 30.2 34.7 35.8 consumptio n

Triglyceride 480 499 530

Placebo 02 31.1 31.2 31.1 consumptio n

Triglyceride 420 418 419

02 consumption as reflected by Nitric Oxide production (nanomoles/ml) and triglyceride concentration (mg%)

Thus, Example 7 increased fat liberation by 17% as measured by changes in plasma triglycerides and increased fat catabolism by 18.5% as measured by increased oxygen consumption.

Example 8

A formulation may be prepared comprising tyrosine about 1,000 mg, glutamine about 1,000 mg, arginine about 500 mg , histidine about 100 mg, cocoa about 800 mg, and sugar about 1,600 mg. These ingredients may be administered as a single dose in the form of a chocolate candy by combining them with melted semi-sweet chocolate with sufficient heavy cream and additional sugar to form an acceptable texture and taste of the resulting chocolate candy. This formulation is preferably administered as in Example 1.

Example 9 A formulation may be prepared comprising tyrosine about 1,000 mg, glutamine about 500 mg, arginine about 500 mg, histidine about 100 mg, cocoa about 800 mg, and dextrose about 1,600 mg. These ingredients may be administered as a single dose in the form of a chocolate candy by combining them with melted semi-sweet chocolate with sufficient heavy cream and additional sugar to form an acceptable texture and taste of the resulting chocolate candy. This formulation is preferably administered as in Example 1.

As can be seen from the foregoing, the synergistic combinations of the invention allow reduced doses of the individual components, particularly of the neurotransmitter precursors and sympathomimetic agents, to be used to achieve the desired effects. The reduced doses decrease side effects caused by the large doses heretofore necessary to achieve the desired effects., Our invention allows mobilization and catabolism of stored fats to be achieved without requiring the use of pharmaceuticals. Our invention allows these desired effects to be achieved at dosage levels of neurotransmitter precursors that are considered safe by regulatory authorities. Previous attempts to use certain of the components in isolation were either ineffective or required dosages which caused side effects.

Although the description above contains many specificities, these should not be construed as Umiting the scope of the invention but as merely providing illustrations of some of the presently preferred embodiments of this invention. Various other embodiments and ramifications are possible within it's scope.

Claims

1. A composition promoting the mobilization and catabolism of lipids in a subject which comprises at least one lipolytic agent selected from xanthines, indirect sympathomimetic amines and precursors for norepinephrine and histamine, in an amount sufficient to increase mobilization of lipids is adipose tissue, and a thermogenic agent comprising a substrate for an endogenous energy-consuming cycle selected from the amino acids glutamine, glutamic acid, aspartic acid, glycine, citrulline, ornithine and their physiologically acceptable salts, hydrates, acid adducts and mineral chelates, in an amount effective to enhance catabolysis of lipids.

2. A composition as in claim 1 and wherein the lipolytic agent comprises a norepinephrine precursor selected from tyrosine, phenylalanine and their physiologically acceptable salts, hydrates, add adducts and mineral chelates, in an amount to mobilize adipose tissue lipids in the subject.

3. A composition as in either of claims 1 and 2 and wherein the lipolytic agent further comprises a histamine precursor selected from histidine and its physiologically acceptable salts, hydrates, add adducts and mineral chelates, in an amount to enhance mobilize adipose tissue lipids in the subject.

4. A composition as in a either claims 2 and 3 and wherein the lipolytic agent comprises a xanthine in an amount sufficient to enhance mobilization of adipose tissue lipids in the subject.

5. A composition as in claim 4 and wherein the xanthine comprises theobromine.

6. A composition as in claim 5 in unit dosage form and wherein the theobromine comprises from about 1 mg, to 2 grams per dose.

7. A composition as in claim 4 and wherein the xanthine is in the form of cocoa.

8. A composition as in claim 4 and wherein the xanthine comprises caffeine.

9. A composition as in any of claims 2 through 8 and wherein the lipolytic agent further comprises an indirect sympathomimetic amine in an amount sufficient to further enhance mobilization of adipose tissue lipids in the subject.

10. A composition as in either claim 1 or 8 and wherein the indirect sympathomimetic amine is selected from epinephrine, ephedrine, pseudoephedrine, isoproterenol, metaproterenol, phentermine and their physiologically acceptable salts, hydrates, add adducts and mineral chelates.

11. composition as in any of claims 2 through 10 and wherein the substrate comprises an amino add selected from glutamine, glutamic add, aspartic add, and their physiologically acceptable salts, hydrates, add adducts and mineral chelates, in an amount effective to enhance catabolysis of lipids.

12. composition as in any of claims 2 through 11 and wherein the substrate comprises an amino add selected from ornithine and its physiologically acceptable salts, hydrates, add adducts and mineral chelates, in an amount effective to enhance catabolysis of lipids.

13. A composition as in any of claim 1 through 12 and wherein the composition further comprises an amino add selected from arginine and its physiologically acceptable salts, hydrates, add adducts and mineral chelates in an amount up to about 1 gram effective to enhance catabolysis of hpids and to promote the release of glucagon and thereby promote mobilization of adipose tissue lipids.

14. A composition as in any of claims 1 through 13 in unit dosage form and wherein the composition further comprises an amount of up to about 3 grams of dextrose equivalents of mono- and/ or di-saccharides sufficient to promote the release of insulin and whereby to effect a drop in blood sugar to promote the release of glucagon and thereby the mobilization of adipose tissue lipids.

15. The composition as in any of claims 1 through 14 in unit dosage form which comprises less than 6 grams of carbohydrates per dose and an amount of amino adds other than said substrate amino adds equal to less than about 50% by weight of the substrate amino adds in the composition.

16. A composition as in any of claims 1 through 15 in unit dosage form and wherein the composition further comprises an amount of up to about 3 grams of dextrose equivalent of mono- and/ or di-saccharides sufficient to promote the release of insulin and thereby to effect a drop in blood sugar to promote the release of glucagon and thereby the mobilization of adipose tissue lipids.

1. A composition promoting the mobilization and catabolism of lipids in a subject which comprises at least one lipolytic agent selected from xanthines, indirect sympathomimetic amines and precursors for norepinephrine and histamine, in an amount sufficient to increase mobilization of lipids in adipose tissue, and a thermogenic agent comprising a substrate for an endogenous energy-consuming reaction selected from the amino acids glutamine, glutamic acid, aspartic acid, glycine, citrulline, ornithine and their physiologically acceptable salts, hydrates, acid adducts and mineral chelates, in an amount effective to enhance catabolysis of lipids.

2. A composition as in claim 1 and wherein the lipolytic agent is selected from indirect sympathomimetic amines and precursors for norepinephrine.

3. A composition as in claim 2 and wherein the lipolytic agent comprises a norepinephrine precursor selected from tyrosine, phenylalanine and their physiologically acceptable salts, hydrates, add adducts and mineral chelates, in an amount to mobilize adipose tissue lipids in the subject.

4. A composition as in claim 3 and wherein the lipolytic agent further comprises an indirect sympathomimetic amine in an amount sufficient to further enhance mobilization of adipose tissue lipids in the subject.

5. A composition as in claim 4 and wherein the indirect sympathomimetic amine is selected from epinephrine, ephedrine, pseudoephedrine, isoproterenol, metaproterenol, phentermine and their physiologically acceptable salts, hydrates, add adducts and mineral chelates.

6. A composition as in claim 2 and wherein the lipolytic agent further comprises a histamine precursor selected from histidine and its physiologically acceptable salts, hydrates, add adducts and mineral chelates, in an amount to enhance mobilize adipose tissue lipids in the subject.

7. A composition as in claim 6 and wherein the Upolytic agent further comprises a xanthine in an amount sufficient to enhance mobilization of adipose tissue lipids in the subject.

8. A composition as in claim 7 and wherein the xanthine comprises theobromine.

9. A composition as in claim 2 and wherein the lipolytic agent further comprises a xanthine in an amount sufficient to enhance mobilization of adipose tissue lipids in the subject.

10. A composition as in claim 9 and wherein the xanthine comprises theobromine.

11. A composition as in claim 10 in unit dosage form and wherein the theobromine comprises from about 1 mg. to 2 grams per dose.

12. A composition as in claim 9 and wherein the xanthine is in the form of cocoa.

13. A composition as in claim 9 and wherein the xanthine comprises caffeine.

14. A composition as in claim 2 and wherein the substrate comprises an amino add selected from glutamine, glutamic add, aspartic add, and their physiologically acceptable salts, hydrates, add adducts and mineral chelates, in an amount effective to enhance catabolysis of lipids.

15. A composition as in claim 2 and wherein the substrate comprises an amino add selected from ornithine and its physiologically acceptable salts, hydrates, add adducts and mineral chelates, in an amount effective to enhance catabolysis of lipids and to promote the release of glucagon and thereby promote mobilization of adipose tissue lipids.

16. A composition as in claim 1 and wherein the composition further comprises an amino add selected from arginine and its physiologically acceptable salts, hydrates, add adducts and mineral chelates in an amount up to about 1 gram effective to enhance catabolysis of lipids and to promote the release of glucagon and thereby promote mobilization of adipose tissue lipids.

17. A composition as in any of claims 1 in unit dosage form and wherein the composition further comprises an amount of up to about 3 grams of dextrose equivalents of mono- and /or di-saccharides sufficient to promote the release of insulin and whereby to effect a drop in blood sugar to promote the release of glucagon and thereby the mobilization of adipose tissue lipids.

18. The composition as claim 1 in unit dosage form which comprises less than 6 grams of carbohydrates per dose and an amount of amino acids other than said substrate amino acids equal to less than about 50% by weight of the substrate amino acids in the composition.

19. A composition as in claim 1 in unit dosage form and wherein the composition further comprises an amount of up to about 3 grams of dextrose equivalent of mono- and/ or di-saccharides sufficient to promote the release of insulin and thereby to effect a drop in blood sugar to promote the release of glucagon and thereby the mobilization of adipose tissue lipids.

20. A method of promoting the mobilization and catabolism of lipids in a subject which comprises concomitantly administering to the subject, at least one lipolytic agent selected from xanthines, indirect sympathomimetic amines and precursors for norepinephrine and histamine, in an amount sufficient to increase mobilization of lipids is adipose tissue, and a thermogenic agent comprising a substrate for an endogenous energy- consuming cycle selected from the amino adds glutamine, glutamic add, aspartic add, glycine, citrulline, ornithine, and their physiologically acceptable salts, hydrates, add adducts and mineral chelates, in an amount effective to enhance catabolysis of lipids.

21. A method as in claim 20 and wherein the lipolytic agent is selected from indirect sympathomimetic amines and precursors for norepinephrine.

22. A method as in either claim 20 or 21 and wherein the lipolytic agent comprises a norepinephrine precursor selected from tyrosine, phenylalanine and their physiologically acceptable salts, hydrates, add adducts and mineral chelates, in an amount to mobilize adipose tissue lipids in the subject.

23. A method as in any of claims 20, 21 and 22 and wherein the lipolytic agent further comprises a xanthine in an amount to enhance mobUization of adipose tissue lipids in the subject.

24. A method as in claim 23 and wherein the xanthine comprises theobromine.

25. A method as in any of claim 20 through 24 and wherein the lipolytic agent further comprises a histamine precursor selected from histidine and its physiologically acceptable salts, hydrates, add adducts and mineral chelates, in an amount to enhance mobilization of adipose tissue lipids in the subject.

26. A method as in any of claim 21 through 25 and wherein the substrate comprises an amino add selected from glutamine, glutamic add, aspartic add, and their physiologically acceptable salts, hydrates, add adducts and mineral chelates, in an amount effective to enhance catabolysis of lipids.

27. A method as in any of claims 21 through 26 and wherein the substrate comprises an amino add selected from ornithine and its physiologically acceptable salts, hydrates, add adducts and mineral chelates, in an amount effective to enhance catabolysis of lipids and to promote the release of glucagon and thereby promote mobilization of adipose tissue lipids.

28. A method as in any of claims 20 through 27 and wherein the composition further comprises an amino add selected from arginine and its physiologically acceptable salts, hydrates, add adducts and mineral chelates in an amount up to about 1 gram effective to enhance catabolysis of Upids and to promote the release of glucagon and thereby promote mobilization of adipose tissue lipids.

29. A method as in any of claims 20 through 28 and including the concomitant administration to the subject of an amount of up to 3 grams of dextrose equivalent of mono- and/ or di-saccharides sufficient to promote the release of insulin and thereby to effect a drop in blood sugar to promote the release of glucagon and thereby the mobilization of adipose tissue lipids.

30. A method as in any of claims 20 through 29 wherein the lipolytic agent and the substrate are administered to the subject on an empty stomach and including the further step of restricting the concomitant intake by the subject of amino adds, other than said substrate amino acids, to an amount less than 50% by weight of the substrate amino acids administered and of carbohydrates, to an amount of less than 6 grams.