CN87106235A

CN87106235A - Methods and matrix compositions for treating atherosclerosis

Info

Publication number: CN87106235A
Application number: CN198787106235A
Authority: CN
Inventors: 斯坦利·J·杜德里克
Original assignee: Dudrick Medical Research Fund 1 Ltd (us) 3600 Allied Bank Plaza Houston Texa
Current assignee: Dudrick Medical Research Fund 1 Ltd (us) 3600 Allied Bank Plaza Houston Texa
Priority date: 1986-09-10
Filing date: 1987-09-10
Publication date: 1988-05-18
Also published as: AU8033487A; DK253688D0; WO1988001872A1; DK253688A; IL83822A0; CA1290692C; KR880701558A; ZA876782B

Abstract

The invention provides a kind of method for preparing base composition, compositions can reduce plasma cholesterol in fact in order to the treatment atherosclerosis, can reverse arterial plaque dirt deposition and the vascular degeneration relevant with atherosclerosis.The step of preparation method comprises the amino-acid mixed pH value that lumps together and regulate compositions within human blood pH allowed band of biological activity.Base composition also comprises biological activity Aminosteril KE solution as antiatherosclerotic.

Description

Methods and matrix compositions for treating atherosclerosis

The present invention relates to compositions and methods for treating atherosclerosis, and more particularly to methods and matrix compositions for treating atherosclerosis that address many of the various problems associated with disease, substantially alleviating symptoms and even curing the disease to some extent. The present invention uses bioactive L-amino acids (L-form) to achieve the purpose.

Cardiovascular disease and its associated diseases, disorders and complications are the leading causes of disability and death in the united states and western europe. One particular disease that is clearly pathophysiologically related is atherosclerosis, which is generally believed to be responsible for the increased mortality and expense of health. The number of people who die of coronary artery disease alone each year in the united states in the past five years has exceeded 550,000. In addition, the number of hospitalizations for myocardial infarction, a complication of atherosclerosis, is also over 680,000 annually, and is steadily increasing. In recent years, the number of deaths from atherosclerosis in the united states has exceeded the sum of deaths from cancer, trauma and infectious disease.

In 1983, the cost of direct health management for patients with coronary heart disease in the united states exceeded $ 80 million. The total cost of atherosclerosis alone in coronary heart disease in this country has been over $ 600 billion at the same time, and is projected to reach $ 960 billion in 1987. These costs have not even reflected the costs for dealing with many different non-cardiac conditions that are also caused by atherosclerosis.

The disease of atherosclerosis is characterized by the deposition and subsequent fibrosis of fatty substances (primarily cholesterol) in the arterial vascular lining, resulting in the deposition and degeneration of plaque on the surface of the arterial wall. The ubiquitous arterial fatty plaque, a thick fat, lipid sebaceous gland follicle composed of macrophages (white blood cells) and some smooth muscle fibers, is an early lesion of atherosclerosis (a condition often found even in children). Plaque fibrosis represents a late-stage progression change in atherosclerosis that allows the smooth muscle cells of the intima of blood vessels to become engulfed by the connective tissue matrix and contain variable amounts of intracellular and extracellular lipids. This dense fibrous cap, usually covered by smooth muscle or connective tissue, is located on the luminal surface of the artery. Beneath the fibrous cap, the lesion is a high cell body consisting of macrophages, other leukocytes and smooth muscle cells, and deep in the region of this massive cell accumulation is a region of cholesterol crystals, necrotic debris and calcified material.

If the disease continues to progress, it causes narrowing and obstruction of the arterial lumen, resulting in reduced or occluded blood flow, which in turn causes ischemia or infarction of major organs or anatomical sites, such as the brain, heart, intestine or limbs. The result is loss of function, loss of cellular material, use of emergency medicine and/or surgery, and can result in disability or even death. In addition, lipids (cholesterol), inflammatory leukocytes, connective tissue and calcifications infiltrate the muscle layer, severely weakening the arterial vessel wall, causing softening and/or embrittlement of certain parts of the vessel wall, forming partial dilatation (hemangioma) and rupture, resulting in organ, limb bleeding and even life-threatening.

The cause of atherosclerosis is not fully understood and it is theorized from the experience of clinical observations by many researchers and clinicians that this disorder is related to plasma cholesterol and lipid content. Generally preferred means of treating atherosclerosis include: medication, diet, change of mode of action and exercise to control and reduce plasma cholesterol levels.

Drugs commonly used to lower plasma cholesterol levels include clofibrate, gemfibrozil, sodium d-thyroxine, colestipol, probucol, niacin and cholestyramine. Generally, these drugs are only etiologically directed treatments and do not contribute to the outcome of atherosclerosis. And has no effect on reversing plaque deposits and reversing degeneration in arterial vessel walls. These agents also have a number of other disadvantages, such as: adverse side effects (hypertension, arrhythmia, gastrointestinal discomfort, headache, allergies, etc.), contraindications (heart disease, liver or kidney disease, pregnancy, etc.), the need to take drugs carefully for life, the patient consistently achieving this is difficult, the duration often changes, and the cost of the drugs is high.

Once the condition has progressed to a significant stage where symptoms persist and cardiac function is compromised, the next treatment is usually an arterial bypass graft to repair and/or replace the diseased artery. While coronary artery bypass surgery has become one of many common cardiovascular surgical procedures in the united states, it is clear that surgery does not address the pathological problem, as it does not serve as a stop or reverse in the progression of the disease, but rather only makes it possible to transiently overcome the most critically ill diseased artery by bypassing the artery when necessary. Further, surgery that patients have had marginal acceptance is associated with significant risk of morbidity and mortality. Indeed, the condition may continue to progress as the surgery progresses, and the veins or arteries themselves often cross the diseased artery after the surgery, which causes atherosclerosis at a faster rate than the original diseased artery. In addition, the coronary surgery study sponsored by the national institute for heart, lung and blood (NHLBI) concluded that there was no statistical significance in survival or rate of progression to myocardial infarction between patients treated surgically and with drugs.

Another approach to coronary bypass surgery is to treat the consequences of atherosclerosis, rather than the cause of the disease, with some drugs and methods. These include chelation therapy with ethylenediaminetetraacetic acid, EDTA, and coronary angioplasty with a percutaneous transluminal metastasis (PTCA). However, EDTA therapies are still in the trial and their benefits and adverse effects are not fully understood and yet to be further appreciated. PTCA therapy is limited in its use by its harmfulness, which sometimes results in fatal complications. The application of advanced experimental intra-arterial laser beam spot mist vaporizers is limited because it requires open surgery to access the diseased vessel.

It has now been unexpectedly found that parenteral administration of solutions containing certain mixtures of biologically active amino acids can significantly reduce plasma cholesterol levels (concentrations) and prevent and reverse arterial plaque deposition and the development of the vascular degeneration associated with atherosclerosis.

For a variety of medical reasons, some solutions of amino acids have been used in the parenteral veins for the past thirty to forty years.

For Total Parenteral Nutrition (TPN) or intravenous eutrophication, there are widely available techniques to address the nutritional and metabolic problems of patients who cannot or should not ingest and absorb sufficient optimal nutritional matrix through the digestive tract. Indeed, the technology created by the present inventors has been successfully used to support the normal development and growth of infants and children over the age of fifteen.

Currently, there are many standard TNP solutions commercially available in the United states, including the TPN solution TRAVOSOL (available from Travenol laboratories), EREAM INE (available from Mc Gaw laboratories), under the trade name TPN. AM INO SYN (manufactured by Abbott laboratories) and NO-VAM INE (Cutter Medical, Divisian of Miles laboratories). These standard intravenous nutritional solutions are nutritionally balanced and formulated to provide the usual patient needs of nutritional solutions containing calories, proteins (amino acids), vitamins, minerals, water and trace elements. The components of the solutions can be used in a certain proportion, and can be prepared into special solutions by doctors and pharmacists according to the special needs of patients in the course of treatment, in order to meet the special nutritional needs of the patients, the essential fatty acids and the increased calories can also be used for patients respectively by veins as the reinforced emulsion of soybean or sunflower seed oil in the form of lipid. For patients with renal disease, liver disease or severe symptoms, specialized intravenous formulations of amino acids are also commercially available which are treated by the addition of adjuvants. For example, U.S. Pat. No. N_o3,832,456 describes the use of amino acids as nutrients, U.S. patent N_o3, 950, 529 describes amino acids for use by patients with liver disease.

However, none have suggested that solutions of biologically active amino acids administered parenterally can be used to treat atherosclerosis, and in particular to reverse the harmful complications associated with the disease.

It is therefore an object of the present invention to provide a method for preparing an amino acid formulation for the treatment of atherosclerosis, which formulation not only lowers plasma cholesterol levels (concentration) (causing pathogenesis), but also inhibits, reverses and even cures to some extent plaque deposits and degenerative arteries (outcome) and secondary complications associated with the disease.

The present invention also provides a method for preparing an amino acid formulation for treating atherosclerosis which does not produce many of the side effects of conventional drug therapy and which is effective in reducing or even eliminating the surgical or other surgical treatment regimen used by most atherosclerotic patients.

The invention provides a formulation for the preparation of a medicament for the treatment of atherosclerosis, characterized in that it comprises: fatty substances are deposited in the inner layers of arteries and fibrosis leads to plaque deposits and degeneration of the arterial wall. The formulations of the present invention have the particular advantage that they can both significantly reduce plasma cholesterol levels and inhibit, reverse or even to some extent cure arterial plaque deposits and vascular wall degeneration associated with atherosclerosis.

In particular, the present invention provides a process for the preparation of an anti-atherosclerotic agent capable of lowering plasma cholesterol levels and reversing atherosclerosis-associated arterial plaque deposits and vascular degeneration, comprising a biologically active amino acid, which is prepared by the steps of:

(A) combining the following:

3% to 10% isoleucine

3% to 10% leucine

About 2.5% to about 7.5% lysine

About 2.5% to 7.5% methionine

About 3% to about 10% phenylalanine

About 1% to about 4% threonine

About 1% to about 3% tryptophan

About 3% to about 10% valine

About 2% to about 20% alanine

About 7% to about 20% arginine

About 1% to about 4% histidine

About 0% to about 4% proline

About 0% to about 8% serine

From about 0% to about 1% tyrosine

About 0% to about 10% glycine

About 0% to about 6% glutamic acid

About 0% to about 6% aspartic acid

About 0% to about 1% cysteine

The weight percentage of the bioactive amino acid is the weight percentage of the whole bioactive amino acid, and the amino acid is preferably selected from acetate, chloride and hydrochloride crystal;

(B) balancing the ions used in the salt.

(C) Non-toxic organic compounds are added to adjust the pH of the formulation to within the pH range of human blood.

The invention also provides a process for preparing an aqueous solution of an anti-atherosclerotic agent for parenteral administration.

In accordance with the general concept, the formulations of the present invention comprise, at least in part, an active anti-atherosclerotic agent that additionally comprises a combination of biologically active (L-form) amino acids. The amino acid formulation preferably contains at least arginine and branched chain amino acids, isoleucine, leucine and valine, wherein arginine is preferably present in the mixture in a maximum weight percentage of the total amino acid weight. Preferred formulations also include lysine, methionine, phenylalanine, threonine, tryptophan, and histidine. The most preferred formulations include alanine, proline, serine, tyrosine, glycine, glutamic acid, aspartic acid and cysteine.

The formulation of the method may also include other amino acids, other nutrients or additives, such as vitamins, minerals, electrolytes, carbohydrates, antibiotics, anticoagulants, etc., depending on the particular needs of the patient.

The compositions of the present method are preferably administered parenterally as an aqueous solution through a central venous catheter, since suitable aqueous solutions are hypertonic. Preferably, a metered amount of the aqueous solution is continuously and constantly administered by intravenous infusion from a vein via an intravenous infusion pump.

The main advantages of the product of the method for treating patients with atherosclerosis and containing other compositions include the following:

the plasma cholesterol levels can be reduced more rapidly and in a greater extent than achieved by any prior drug treatment, the extent and range of atherosclerotic plaque regression is greater than previously shown, the treatment time required to reverse atherosclerotic plaque is relatively short compared to plaque growth from baseline, all solution components are biologically active nutrients with negligible toxicity and adverse reactions when administered within the prescribed dosage range, the former is more advantageous in treating atherosclerosis than the latter in terms of cost-effectiveness-benefit ratio when compared to other drugs and surgery, the treatment can be initiated and completed to a portal patient, and the method treats and addresses the underlying pathophysiology of atherosclerosis by an initial and biochemical route for therapeutic purposes rather than by a pharmacological or surgical route involving only atherosclerotic complications.

These and other features and advantages of the present invention will be readily apparent to those skilled in the art from the following detailed description and the accompanying drawings.

Figure 1 shows the daily total plasma cholesterol concentrations in patients treated as described in example 2, showing the response obtained by varying the composition of the infusion.

In accordance with the present method, there is provided a substrate composition for the treatment of atherosclerosis, the aforementioned disorders characterized by fatty substances, initial cholesterol as plaques and subsequent fibrotic deposition on the inner layers of arteries. According to the description, atherosclerosis is believed to be at least partially related to plasma cholesterol and lipid levels.

The compositions and methods of the present invention are particularly effective in reducing plasma cholesterol levels (contributing factors) and in alleviating and to some extent curing the disease by reversing arterial plaque deposition and vascular wall degeneration (outcome), respectively, and are particularly effective in addressing many of the problems, conditions and complications associated with the disease.

In accordance with the general concept, the compositions of the present invention comprise a biological substrate that includes, at least in part, an anti-atherosclerotic agent. The anti-atherosclerotic agent used herein has effects of lowering plasma cholesterol level and inhibiting and reversing arterial plaque deposition and arterial wall degeneration. In particular, the active anti-atherosclerotic agent is of biologically active amino acids, especially of the L-form.

Biologically active amino acids are generally classified as essential and non-essential. Essential amino acids are those which cannot be synthesized in vivo and which must be taken up in sufficient quantities from food. Non-essential amino acids are those that can be biosynthesized in vivo using other substrates. Amino acids that are generally classified as essential are: isoleucine, leucine, and lysine, methionine, phenylalanine, threonine, tryptophan, and valine. Non-essential amino acids are: alanine, arginine, histidine, proline, serine, tyrosine, glycine, glutamic acid, aspartic acid, and cysteine. Arginine and histidine are considered to be semi-essential in that they are essential for infants and to some extent also for renal patients.

These biologically active amino acids are dextrorotatory D-form and levorotatory L-form. L-form is believed to be more biologically active than D-form, and therefore L-form is preferred for use in anti-atherosclerotic formulations.

The substrate compositions of the present invention are placed in an aqueous solution and then administered to a patient parenterally as a therapeutic agent. When this administration is undertaken, the compositions of the present invention do reduce plasma cholesterol levels and reverse arterial plaque deposition and vessel wall degeneration.

Preferably, the anti-atherosclerotic agent comprises a mixture of at least arginine, isoleucine, leucine and valine, wherein arginine is preferably the greatest weight percentage of the total bioactive amino acids in the mixture. These amino acids preferably comprise at least 25% by weight of the amino acids, but may be the only active ingredient when used in a short infusion cycle. The mixture also includes various other amino acids. Preferably all of the above mentioned amino acids are present in the mixture so that all amino acids are used together with the other ingredients in the nutritional balancing solution. All of these components are part of the TPN solution, but unexpectedly increasing the branched-chain amino acid and arginine concentrations, usually accompanied by a decrease in the amount of histidine in the amino acid mixture, results in a significant (typically 40% to 60%) decrease in plasma cholesterol levels in time (about one week). Another note is the reduction of simple amino acids in the TPN solution, especially alanine and glycine, especially glycine.

The amino acids contained in the preferred amino acid mixture and their approximate weight percent ranges based on the weight of the total amino acids in the mixture are set forth in Table I below.

TABLE I

Amino acid about weight%

Isoleucine 3-10 alanine 2-20

Leucine 3-10 histidine 1-4

Lysine 2.5-7.5 proline 0-10

Methionine 2.5-7.5 serine 0-8

Phenylalanine 3-10 tyrosine 0-1

Threonine 1-4 Glycine 0-10

Tryptophan 1-3 glutamic acid 0-6

Valine 3-10 aspartic acid 0-6

Arginine 7-20 cysteine 0-1

It should be noted that the nonessential amino acids listed above as optional components in the broad sense of the invention have less effect on the recovery from atherosclerosis than other classes of amino acids, as are alanine and amino acids without a branch. However, these amino acids are important from a nutritional point of view when the patient requires months of treatment. In light of the above, the addition of more or less of these amino acids than listed in the table above will not affect or alter the properties and advantages of the solution and, therefore, will not affect the practice of the present invention. Thus, the following amino acids contained in the preferred amino acid mixture and their approximate weight percent ranges based on the weight of the total amino acids in the mixture are set forth in Table II below.

TABLE II

Approximate weight percent amino acids

Isoleucine 4-10 alanine 7-16

Leucine 5-10 histidine 1.5-3.5

Lysine 4.5-6.5 proline 4.5-3.5

Methionine 3.5-5.5 serine 2.5-8.5

Phenylalanine 4.5-7.5 tyrosine 0-1

Threonine 2-3.5 Glycine 3-8

Tryptophan 1-2.5 glutamic acid 3.5-5.5

Valine 4-9 aspartic acid 2-4

Arginine 9-18 cysteine 0-1

The above amino acid mixture is preferably prepared by mixing together the amino acids in crystalline form, usually as acetate, chloride or hydrochloride. The salt is balanced to avoid disruption of the acid-base balance in the patient, and usually about 80% acetate, 20% chloride and hydrochloride are used. The pH of the solutions of the present invention may generally range from about 5.0 to about 7.8, but is preferably adjusted to the same pH as blood, i.e., about 7.39, by the addition of a suitable non-toxic organic compound. Crystalline forms of amino acids are chosen because of their high purity and because the corresponding amino acids can be accurately regenerated from the crystalline forms of amino acids. Furthermore, solutions of amino acids in crystalline form can be directly assimilated in vivo, and daily biological action is required in vivo. The amino acid mixture may also be formulated using commercially available parenteral nutrition (TPN) solutions to which the amino acid is added, preferably in crystalline form, to obtain the solutions of the invention. Useful TPN fluids include those available under the trade designation "Travasol".

In addition, the compositions of the present invention may contain other essential nutrients, such as vitamins, minerals, electrolytes, carbohydrates, antibiotics, anticoagulants, etc. that have a nutritional, antibacterial, anticoagulant effect, as well as other bio-supporting substances known to those skilled in the art. However, the optimum composition will depend on the particular patient, which is the same in almost all areas of biological use, i.e., the amino acid mixture actually used will be adjusted based on factors that generally determine the composition and dosage to be administered, the tolerance of the particular patient to the individual and combination of ingredients, the stage of disease progression and extent of disease, the major organ(s) or system(s) damaged by the disease and/or its complications, the major biological functions damaged by the disease and/or its complications, the patient's concomitant malnutrition, the presence, nature and intensity of clinical syndromes, the recovery stage and the stage following surgery or other procedures to reduce the signs and symptoms of the disease and/or its complications, the age of the patient, Gender and general condition, the development of other concomitant diseases (diabetes, familial hyperlipidemia, etc.), and other factors known to those skilled in the art.

These factors are related to the ability of the patient to use or tolerate the amino acid mixture and other ingredients of the composition of the invention. In other words, due to one or more of the above listed factors, some adjustment of specific components may be required for each patient at different stages of treatment. For example, a patient with diabetes may not tolerate as much of the thermogenic component in the glycoform as a patient without diabetes. As another example, it is known that the amino acid requirements of a person with renal or hepatic failure in a patient are different from those of a person without such symptoms. As another example, a person who is malnourished may require more and different amino acids and other nutrients than a person who is well-nourished. Those skilled in the art will recognize these particular needs and will be able to adjust the compositions of the present invention accordingly.

Other adjustments to the amino acid mixture may also be made based on the blood or urine amino acid profile. These profiles reflect the body's utilization of each specific amino acid, from which profile it can be determined which amino acid is lacking in the body, or alternatively. That amino acid is in excess and can therefore be compensated by adjusting the amino acid mixture described above. Such amino acid profiles can be adjusted as desired by those skilled in the art once they have been determined. The condition of the body of the patient using the amino acid is determined by blood and urine analysis, and the amino acid distribution is adjusted according to the analysis result. We have found that the anti-atherosclerotic effect is largely dependent on the amount of branched chain amino acids and arginine used in the treatment. To date, the particular significance of arginine as an important amino acid has been overlooked, and has been unexpectedly revealed in the practice of the present invention: arginine is an important contributing amino acid.

The compositions of the present invention are preferably administered parenterally in the form of an aqueous solution for intravenous injection. The aqueous solution thus contains the above-described composition as a solute added to the solution and an aqueous solvent base. The aqueous solvent base is preferably a sterile deionized water base to which several ingredients may be added depending on the composition of the amino acid mixture and the specific needs of the individual patient. For example, if the caloric value of the composition is insufficient and the patient needs to take more calories, the aqueous solvent base can be glucose-water, fructose-water, or invert sugar-water (containing equal amounts of glucose and fructose), containing varying concentrations of carbohydrate substrates selected to meet the patient's caloric requirements. The above solutions can be formulated using sterile, aseptic and pharmaceutical chemical methods well known to those skilled in the art that have been shaped to ensure complete dissolution of the ingredients, while maintaining stability, sterility, safety and to ensure a better formulation for the individual patient.

The solution preferably contains an effective amount of an anti-atherosclerotic agent, i.e. a mixture of amino acids, to produce the following effect in a relatively short period of time, typically two to six months, (during which time the patient is supplied with nutrients necessary for life support) following parenteral administration of the solution by a hypertrophic method well known to those skilled in the art: the plasma cholesterol is obviously reduced, and the process of the plaque deposition of the artery and the degeneration and change of the artery wall is obviously reversed. The concentration of the amino acid mixture in the aqueous solution may range from as low as only 4 grams of total weight of amino acids per hundred milliliters of solution (4% by weight of solution) up to the solubility limit of the particular amino acid mixture in the particular aqueous solvent (sometimes as high as 20% by weight of solution). The concentration of the mixture of amino acids in the solution is preferably in the range of about 7.5 to about 12 grams (7.5 to 12% by weight of the solution) of total weight of amino acids per hundred milliliters of the solution. A hypertonic solution (800-1100 milliosmoles per liter) can be prepared using the above concentrations, i.e., the solution has an osmotic pressure greater than that of blood (290-310 milliosmoles per liter), the significance of which is discussed below. For the specific effective compositions described above, the concentration of the amino acid mixture in the solution is preferably in the range of about 9 to about 11 grams, more preferably about 10 to about 11 grams, of amino acid per hundred milliliters of solution.

The aqueous solution of the present invention is administered parenterally rather than orally because oral administration has many disadvantages in that amino acids are subject to modified digestion by the stomach, intestines, pancreas and liver after oral administration, the endogenous loops create a buffering effect on the ingredients in the formulation, and it is difficult to model the solution for oral administration to achieve the same concentration of the anti-atherosclerotic active agent in the blood after oral administration as it would be if it were directly titrated intravenously, and it is difficult to establish and maintain a method for accurately controlling the amount of intake. By parenteral administration, the target tissue can be directly attacked and in some cases even isolated and perfused with an amino acid solution for a period of hours, days or weeks without drastically altering the patient's normal nutritional profile. Furthermore, the controlled constant administration can be achieved by parenteral administration, which ensures the duration of the therapeutic effect and thus avoids the great rise in the therapeutic effect, which is inevitable during absorption and assimilation after oral or enteral administration.

The aqueous solution selected may be hypertonic (about 800 to about 1100 milliosmol/liter) so that it is administered intravenously to a large diameter vessel with high blood flow to achieve substantially immediate dilution, preferably by intravenous administration through a central venous catheter which is inserted percutaneously into the superior vena cava by any suitable method known to those skilled in the art. A good catheterization method is described in an article by O' Donnell, Clague and Dudrick entitled "Percutaneous Insertion of a affected cathter with a long Subcutaneous catheterization Tunnel for Intra hyperthermia", published in southern journal of medicine 76, No. 11 (1983), p.1344-1348, the entire contents of which are hereby incorporated by reference.

After catheterization, uninterrupted instillation with an anti-atherosclerotic aqueous solution may begin. The total daily instillation of the solution is an amount sufficient to provide about 0.25 to 3.5 grams, and more preferably about 1 to 2 grams, of amino acid per kilogram of body weight per 24 hours for a single patient. It is also desirable that the daily amount of the above solution is continuously and constantly administered intravenously using a commercially available intravenous pump, so that the amino acid level in the body can be kept constant.

During the course of a continuous infusion therapy, typically about two months to about eight months, it is desirable not to allow the patient to ingest other nutritional agents that affect amino acid balance or metabolism in the body. The patient can drink liquid without nutrient components, and can also take some medicaments which may need to be used in the process of oral treatment. For some particular patients, instillation of the solution may be performed using a compact, lightweight pump and a flow-through device that supports the solution, drip tube and pump, as is done for ambulatory or outpatient patients. Even more particularly, for some patients, it may be preferable to titrate the daily 24-hour daily bolus over 12-16 hours, leaving the patient free of the intravenous pump and infusion set for 8-12 hours per day, allowing them greater freedom and mobility during non-infusion times. When the patient is removed from the infusion tube, in order to prevent the catheter from being blocked by blood clots, it is necessary to inject 1-2 ml of heparin solution containing about 100 units per ml into the central catheter tube, and then to tightly plug it with a sterile plug (e.g., Luer-lock).

The condition of the patient should be monitored biochemically, hematologically, and metabolically through a blood or urine sample, every other three days during the first weeks of infusion of the anti-atherosclerotic solution. Blood and/or urine amino acid profiles of the samples can be made as needed, and the amino acid mixture can then be adjusted according to this profile. If these amino acid indices are stable at the beginning of the infusion of the aqueous solution and any such adjustments to the initial composition of the solution, the monitoring interval may be extended to 1 to 2 weeks.

Other indicators to be monitored include: serum cholesterol, cholesterol ester, apolipoprotein A, apolipoprotein B, High Density Lipoprotein (HDL), cholesterol, triglyceride, sodium, potassium, chloride, bicarbonate, creatinine, calcium, inorganic phosphorus, glucose, urea nitrogen, uric acid, total protein, albumin, and bilirubin, alkaline phosphatase, Lactate Dehydrogenase (LDH), serum glutamic-oxaloacetic transaminase (SGOT), serum glutamic-pyruvic transaminase (SGPT), gamma-glutamyl transferase (GGT), Creatine Phosphokinase (CPK), magnesium, zinc, copper, blood prothrombin time (blood prothrombin time), partial tissue thromboplastin time (partial thromboplastin time), white blood cell count, red blood cell count, hemoglobin, hematocrit, mean cell volume, mean particulate hemoglobin (mean corpuscular hemoglobin), mean particulate hemoglobin concentration, white blood cell differential count, and platelet count. These standard tests can be carried out separately. Grouped or performed together to ensure safety and efficacy of treatment, maintain homeostasis, and further assist clinicians in solution adjustment for individual patients.

The invention has been discussed in detail above, and some specific examples are given below to further illustrate it. These examples are merely illustrative and not restrictive of the invention described above.

Example 1

Attempts to demonstrate that cholesterol is one of the major causes of atherosclerosis have been hampered by the inability to reduce and maintain reduced serum cholesterol levels by either dietary adjustments and/or chemotherapy alone. In a series of preliminary experiments, an animal model of atherosclerosis was developed over a period of several years by adjusting the diet of new zealand albino rabbits. In some studies, hundreds of male and female rabbits were used, and their diets were continuously and logically adjusted, resulting in the development of a clean and reliable set of animal models that could be used to induce atherosclerosis in these albino rabbits, where the induced atherosclerosis was similar in distribution, pathological stages and severity, gross morphological and histological features, basic pathophysiology, secondary complications and clinical manifestations to that observed and documented in human atherosclerosis.

As a herbivore, rabbits generally digest only a small amount of the cholesterol contained in the diet. Plasma cholesterol levels are rapidly increasing due to the inability of the rabbit liver to biochemically clear large amounts of dietary cholesterol. The increased plasma cholesterol concentration in turn leads to a reproducible, higher stage of atherosclerosis over time, the extent, appearance and extent of which is consistent with the increased plasma cholesterol concentration and the duration of hypercholesterolemia.

In the previous study, an atherogenic diet was formulated and used to induce atherosclerosis in hundreds of additional rabbits, which were used as subjects in future studies. In a typical study, 40 New Zealand albino male and female rabbits were fed a basic laboratory rabbit diet (Teklad T.D.82135) consisting of high fiber Purina Labchow 5326 fortified with 8% by weight peanut oil as a carrier for 2% by weight crystalline cholesterol. After the initial feeding of the experimental rabbits, the plasma cholesterol level of the rabbits rose above 1000 mg/dl within ten days to two weeks and was maintained between 1000 and 2000 mg/dl for the remainder of the atherosclerosis-inducing period of six weeks total.

At six weeks later, 10 rabbits were sacrificed and necropsy was performed to delineate and record the extent, stage and extent of progression of atherosclerosis both macroscopically and microscopically observed as a point of reference. The remaining 30 rabbits in the pilot group were randomly divided into three study groups. In the first group, 10 test animals were fed on the atherogenic diet described above for an additional six weeks. A second group of 10 test animals were fed standard test rabbits (Teklad t.d.82135) without peanut oil and crystalline cholesterol for six weeks. A continuous infusion of an anti-atherosclerotic solution was administered to a third group of 10 animals for six weeks via a central venous catheter inserted into the superior vena cava, the method of catheter insertion being known to those skilled in the art. The amino acid composition of the anti-atherosclerotic solution containing 4.25% by weight of amino acids and 25% by weight of glucose is shown in Table III below (column A). The solution is administered in an amount to provide 1.5 grams of amino acids and 35 calories per kilogram of body weight per day, and further comprises vitamins, minerals and trace elements to form a nutritionally balanced mixture.

TABLE III

Amino acid wt%

A B A B

Isoleucine 4.76.3 alanine 2015.8

Leucine 5.97.4 histidine 4.13.2

Lysine 5.96.3 proline 9.48.4

Methionine 5.95.3 serine 7.06.3

Phenylalanine 5.96.3 tyrosine 0.50.4

Threonine 3.53.2 Glycine 9.46.3

Tryptophan 2.42.1 glutamic acid-3.2

Valine 4.76.3 aspartic acid-2.1

Arginine 10.010.5 cysteine 0.70.6

At the end of the second six week study period, 30 rabbits were sacrificed in their entirety and autopsied to determine and trace the severity, stage and distribution of atherosclerosis. In the first group of rabbits, a structurally complex atherosclerotic lesion was induced, which was repeatedly and consistently implicated in 85% to 95% of the aorta leading from the heart to the common iliac branches, with the superficial plaque pattern and arterial wall distribution characteristic of atherosclerotic lesions observed in humans. There were indications that several rabbits in the first group had similar human myocardial infarction, cerebral thrombosis, intestinal ischemia and ileus, and hindlimb ischemia and vascular infarction.

The second group of rabbits fed the standard experimental rabbit diet for six weeks showed no regression of atherosclerotic plaque compared to 10 rabbits sacrificed six weeks after the initial group had been fed the atherogenic diet. In contrast, atherosclerosis in the second group of rabbits also progressed slightly during this second six weeks.

However, significant regression of atherosclerotic plaques occurred in the third group of rabbits that received the vein of the particular anti-atherosclerotic preparation described above. In fact, approximately 90% of the aorta were free of any signs of atherosclerosis, and the concentration of amino acids in plasma and urine was measured every other week in the third group of rabbits, with daily intravenous amino acid supply increasing from 1.5 g per kg body weight to 17 g per kg body weight and adding glutamic acid and aspartic acid to the formulation. The final amino acid composition of the formulation administered to the third group of rabbits is given in column B of table iii. Note that in this composition the branched chain amino acids increased most, while arginine increased slightly. These experiments were repeated until the results of all 200 rabbits in each of the three groups were consistent with the above results.

This experiment shows that the exceptional regression of atherosclerotic plaque is the result of intravenous amino acid treatment alone, which lays the foundation and avoids the loss for further experimental studies on patients.

The study was initially transferred to a trial of patients with the same formulation of anti-atherosclerotic solution injected into the last 40 rabbits in chronological order.

Example 2

A40 year old patient with severe systemic atherosclerosis secondary to familial type II hyperlipidemia, previously treated in a lipid research clinic with a low cholesterol/fat, low calorie (1400 calories) diet in combination with cholecysteamine, is now contemplated for treatment with the solution of the present invention. The plasma cholesterol level in the patient after even 5 months of treatment by the former method was only reduced from the baseline of 496 mg/dl to 409 mg/dl (normal plasma cholesterol level was up to 260) and the patient did not feel any reduction in the symptoms described below. Within two weeks after the patient voluntarily stopped the regimen, the plasma cholesterol level returned to the level of 536 mg/dl at 490-.

One year ago, patients had a recovery of myocardial infarction, but the patients still had heartache, suffered from severe leg cramps and leg lameness after walking only one block, and frequently suffered from stumbling and numbness of the limbs, which is associated with poor blood flow in their major arteries. Typically 4 of the 8 distal pulses palpable on the arms and legs were not palpable, with the remaining 4 pulses having a reduction of about 50%. Coronary angiography indicated complete occlusion of the right major coronary artery secondary to atherosclerosis with diffuse constriction of the left anterior descending and annular coronary arteries and reduced blood flow. There was no significant collateral circulation of the myocardium and he was unable to perform coronary bypass surgery due to the systemic extent of the disease.

His life expectancy was less than two years. The plasma cholesterol concentration was 540 mg/dl. He was only reluctant to walk, and the number and severity of the twitching episodes increased, so he had fallen and injured several times. Magnetic resonance images of the abdominal aorta, iliac artery and carotid artery further confirmed that 80% of the lumens of these vessels were blocked by atherosclerotic plaques. Doppler ultrasound studies of both lower extremities showed a significant reduction in bilateral arterial blood flow.

A percutaneous central venous catheter was inserted into the superior vena cava through the right subclavian vein and the patient was first continuously infused with an intravenous solution having the amino acid composition listed in table iiib above. Initially the patient receives 1.35 g of amino acid per kg of body weight or, that is, 95 g of amino acid per day in the above-mentioned formulation. The patient also received 35 calories of heat per kilogram of body weight in the form of glucose, along with their daily requirements for vitamins, minerals and trace elements, using standard intravenous supplements. Through six months of treatment and clinical studies, some basic metabolic data are gradually obtained. Patients were asked to not take any food other than water throughout the study.

The initial solution formulation was adjusted several times during the course of the anti-atherosclerotic infusion therapy and study in patients based on the plasma cholesterol levels shown in figure 1.

As can be seen in FIG. 1, the plasma cholesterol concentration dropped from the initial 508 mg/min to 450 mg/dl within two days. At point 1, the total daily calories in the mixture were reduced from 2500 calories (35 calories/kg) to 2100 calories (30 calories/kg, which is not expected initially because the patient's activity was less, plasma cholesterol levels further decreased to 370 mg/dl on day eight of treatment, but increased to 393 mg/dl on day nine, at point 2 the daily dosage of the branched chain amino acids lysine, isoleucine and valine was increased by 2 grams, at which point the following amino acids each make up the approximate weight percentages of the total weight of amino acids in the mixture of 7.9% isoleucine, 8.9% leucine, 5.9% lysine, 5.0% methionine, 5.9% phenylalanine, 3.0% threonine, 2.0%, valine, 7.9% alanine, 14.9% arginine, 9.9%, histidine, 3.0% proline, 7.9%, serine, 5.9%, tyrosine, 0.4%, glycine: 5.9%, glutamic acid: 3.0%, aspartic acid: 2.0%, cysteine: 0.6 percent. The daily amount of amino acids per kg body weight was increased from 1.35 g to 1.45, and the total amount of amino acids in the above preparation was 101.5 g per day.

Within 10 days after the start of treatment, plasma cholesterol levels were dramatically reduced to 220 mg/dl, a 57% reduction from the initial level of 508 mg/dl. The concentrations of the various amino acid components and the number of glucose calories were slightly changed at the 3 rd, 4 th and 5 th spots based on the results of biochemical measurements of serum and urine samples. More specifically, at point 3, 1 gram of phenylalanine was added and 1 gram of glycine was decreased in the total daily supply of amino acid salts. At point 4, daily glucose supply was increased to provide 35 calories/kg of calories as the patient's activity increased and weight loss began. At point 5, alanine was reduced by 3 grams, while glutamic acid and aspartic acid were increased by 1.5 grams each, of the total amount of amino acids supplied daily.

At point 6, 10% of the single 500 ml dose was administered by intravenous infusion. After the soybean oil emulsion (sold under the registered trademark Intralipid), the plasma cholesterol level rose dramatically from 260 mg/dl to 370 mg/dl. Points 7 and 8 represent the total heat in the form of glucose given, reduced to 34 cal/kg and 33 cal/kg, respectively. The plasma cholesterol was maintained at a level significantly higher than the minimum level of plasma cholesterol achieved prior to the fat infusion for at least four weeks following the fat emulsion infusion. After the maintenance period, plasma cholesterol again dropped back to 300 mg/dl. At the fourth point, serum plasma cholesterol levels rose to 345 mg/dl after intravenous administration of a single 500 ml dose of 20% soybean oil emulsion (Intralipid), further confirming the deleterious effects on plasma cholesterol even with the administration of essential fatty acids such as soybean oil emulsion. At point 10, daily glucose calories were reduced to 30 calories/kg, while daily doses of proline and serine were reduced by 2 grams each. Since the patient felt a decrease in energy and weight, the daily glucose supply was restored to 35 calories/kg at point 11. At point 12, the concentration of arginine in solution was nearly doubled while the amount of histidine was reduced. The composition of the amino acid mixture at this point, where plasma cholesterol dropped significantly from 306 mg/dl to 245 mg/dl, is shown in Table IV below. The amount of amino acids at this point increased from 1.45 grams per day to 1.5 grams per kilogram of body weight, with a total daily amino acid supply of 103.5 grams.

TABLE IV

Amino acid wt% mol%

Isoleucine 7.77.8 alanine 11.617.0

Leucine 8.78.6 histidine 2.41.5

Lysine 5.84.2 proline 5.86.6

Methionine 4.84.3 serine 3.94.8

Phenylalanine 6.85.3 tyrosine 0.40.3

Threonine 2.93.2 Glycine 4.88.4

Tryptophan 1.91.3 glutamic acid 4.43.9

Valine 7.78.6 aspartic acid 3.43.3

Arginine 16.410.2 cysteine 0.60.6

In the next week, coronary angiography, magnetic resonance images of the patient's aorta, carotid artery and iliac artery, and doppler ultrasound studies of both lower extremities were repeated. Treatment and study were then terminated and patients resumed eating a cholesterol containing diet with a daily cholesterol limit of 100 mg.

During the six month treatment period, patient heartburn or syncope did not occur, three of the four previously untouched terminal pulses were now available, the four previously untouched terminal pulse force increased by 50-100%, the stationary Doppler ultrasound study showed a steady increase in blood flow to the legs to 250-300%, and the patient could walk 3-4 miles per day at a rate of 4 miles per hour without leg cramps or leg lameness.

At the end of the six month treatment period, magnetic resonance images showed a 40-50% reduction in abdominal aorta, common iliac and carotid atherosclerotic plaque. Repeated coronary angiography revealed extensive unobstructed left anterior descending and circumambient coronary arteries, as well as extensive unobstructed multiple collateral arteries leading to the right coronary artery, except for the previous total occlusion at its point of origin.

In addition to the phenomenon that these surprising atherosclerotic plaques have regressed and increased blood flow through the previously damaged multiple arteries has been shown, there is also evidence that the patient's physical function has improved. The patient was able to participate in an 18 hole golf game and completed 18 holes with 70 clubs and was hired as a carpenter at a house building company. He has remained asymptomatic and active for more than 15 months since he started to receive treatment. Periodic observations and studies were continued to confirm the success of the regression of atherosclerosis and the improvement of arterial blood flow and body function (which regression and improvement are already evident in the patient, as described above), and the results showed that the regression and improvement was not previously achievable using any of the medication and dietary treatments.

The final solution developed during the six month treatment study on this patient was then infused into another 10 rabbits, which 10 rabbits were part of a supplemented group of 40 rabbits studied in the same manner as described above for all other things. The atherosclerotic degeneration demonstrated in the 10 rabbits of this supplemented group was most effective, consistent and complete in the atherosclerotic degeneration of the animals studied, and the disappearance rate of atherosclerotic disease was 98% -99%.

Example 3

Additional evidence of the effectiveness of the method was obtained in the context of 30 volunteers who had received a full parenteral nutritional supply for two months to eight years via a central venous feeding catheter and formulated into a composition for the treatment of atherosclerosis. Within one week after changing the aqueous infusion solution (aqueous solution infusion) to the amino acid composition set forth in Table III or Table IV, the plasma cholesterol levels of all subjects were reduced by 40% -60% based on their normal cholesterol levels, regardless of absolute value. These findings were of high significance, as the reduction in plasma cholesterol levels shown in these recipients was orders of magnitude greater, and the positive response rate was 100%. All these patients received prolonged intravenous infusion with amino acid mixtures containing conventional TPN substrates, but without any significant reduction in plasma cholesterol levels. While the infusion compositions of table iv resulted in a significant reduction in plasma cholesterol levels.

Table iv a prominent example of the effect of the solution described in table iv in lowering plasma cholesterol was to lower the patient's plasma cholesterol concentration from an otherwise low baseline concentration of 83 mg/dl to 50 mg/dl within a week, indicating a 40% reduction in plasma cholesterol using an aqueous solution having the amino acid composition listed in table iv.

These data suggest that the mechanism of action of the present invention may be an active biochemical process, rather than the result of strict restriction of dietary cholesterol, cholesterol precursors and other lipids. These data also demonstrate the effectiveness of the present invention. The compositions and methods of the present invention, as described above and in the examples, provide a unique, powerful and effective treatment for atherosclerosis. The recently completed lipid research consultation and primary prevention trial of coronary heart disease by the American society of health organization was the first study in humans to finally demonstrate that lowering blood cholesterol levels can reduce heart attacks and death from heart attacks. However, despite increased dietary control and administration of plasma cholesterol lowering agents, the mean plasma cholesterol level was only reduced by 8.6% during the 10 year study, and the reduction in plasma cholesterol in the best responding patients was not higher than 19%. However, the incidence of myocardial infarction in this study group was also reduced by 19% compared to the general population. The incidence of myocardial infarction can also be reduced more significantly if the plasma cholesterol level is reduced by 40-60%, and with the present invention, as has been previously demonstrated, such an intense reduction in plasma cholesterol level can be achieved.

It should be noted that many modifications and adaptations of the above-described compositions and methods may be made without departing from the spirit of the invention, beyond the embodiments specifically set forth herein. Accordingly, it is to be clearly understood that the present invention has been described in this specification by way of illustration only and is not to be taken as limiting the scope of the invention.

Claims

1. A method for preparing an anti-atherosclerotic agent for reducing plasma cholesterol levels and reversing plaque deposits and vascular wall degeneration associated with atherosclerosis, the agent comprising a biologically active amino acid, the method comprising the steps of:

(A) isoleucine at about 3% -10% and leucine at about 2.5% -7.5% and methionine at about 2.5% -7.5% and phenylalanine at about 3% -10% and threonine at about 1% -4% and tryptophan at about 1% -3% and valine at about 3% -10% and alanine at about 2% -20% and arginine at about 7% -20% and histidine at about 1% -4% and proline at about 0% -10% and serine at about 0% -8% and tyrosine at about 0% -1% and glycine at about 0% -10% and glutamic acid at about 0% -6% by weight based on the total weight of the bioactive amino acids Aspartic acid is combined with about 0% to about 1% by weight cysteine, and the amino acid is preferably in any crystalline form selected from the group consisting of acetate, chloride and hydrochloride salts.

(B) Balancing the ions used in said salt; and are

(C) Non-toxic organic compounds are added to the medicament to adjust the pH to within the human blood pH range.

2. The method of claim 1, wherein said amino acids are combined in the following amounts by weight:

about 4% -10% by weight isoleucine and about 5% -10% by weight leucine and about 4.5% -6.5% by weight lysine and about 3.5% -5.5% by weight methionine and about 4.5% -7.5% by weight phenylalanine and about 2% -3.5% by weight threonine and about 1% -2.5% by weight tryptophan and about 4% -9% by weight valine, and about 7% -16% by weight alanine and about 9% -18% by weight arginine and about 1.5% -3.5% by weight histidine and about 4.5% -8.5% by weight proline and about 2.5% -6.5% by weight serine and about 0% -1% by weight tyrosine and about 3% -8% by weight glycine and 3.5% -5.5% by weight glutamic acid and about 2% -4% by weight aspartic acid and aspartic acid About 0% to about 1% by weight cysteine.

3. The method of claim 1 or 2, wherein the amount of arginine added is the greatest weight percent of the total weight of said biologically active amino acids.

4. The method of claim 1, 2 or 3 wherein said biologically active amino acids comprise their left-rotated form.

5. A method of preparing a composition comprising an active anti-atherosclerotic agent and for treating atherosclerosis, comprising the steps of:

(A) at least the following amino acids isoleucine, leucine, valine and arginine in the biological active amino acids are combined in effective dose, and

(B) adding a non-toxic organic compound to adjust the pH of said composition within the pH range of human blood;

parenterally administering said composition in the form of an aqueous solution reduces plasma cholesterol levels, reverses plaque deposits and vessel wall degeneration associated with atherosclerosis during an effective period, and arginine is added to said composition in a maximum amount by weight based on the total weight of said bioactive amino acids.

6. The method of claim 5, wherein lysine, methionine, phenylalanine, threonine, tryptophan and histidine are further added to said composition.

7. The method of claim 5 or 6, wherein alanine, proline, serine, tyrosine, glycine, glutamic acid, aspartic acid and cysteine are further added to the composition.

8. The method of claim 5, 6 or 7 wherein said biologically active amino acids are selected from the group consisting of their levorotatory crystalline forms.

9. The method of claim 7, wherein said amino acid is added to said solution in the following amounts:

about 3.5% to about 10% by weight isoleucine, 3% to about 10% by weight leucine, 2.5% to about 7.5% by weight lysine, 2.5% to about 7.5% by weight methionine, 3% to about 10% by weight phenylalanine, 1% to about 4% by weight threonine, 1% to about 3% by weight tryptophan, 3% to about 10% by weight valine, 2% to about 20% by weight alanine, 7% to about 20% by weight arginine, 1% to about 4% by weight histidine, 0% to about 10% by weight proline, 0% to about 8% by weight serine, 0% to about 1% by weight tyrosine, 0% to about 10% by weight glycine, 0% to about 6% by weight glutamic acid, 0% to about 6% by weight aspartic acid, 0% to 1% by weight of cysteine.

10. The method of claim 9, wherein said amino acid is added to said solution in the following amounts by weight:

about 4% -10% by weight isoleucine and about 5% -10% by weight leucine and about 4.5% -6.5% by weight lysine and about 3.5% -5.5% by weight methionine and about 4.5% -7.5% by weight phenylalanine and about 2% -3.5% by weight threonine and about 1% -2.5% by weight tryptophan and about 4% -9% by weight valine and about 7% -16% by weight alanine and about 9% -18% by weight arginine and about 1.5% -3.5% by weight histidine and about 4.5% -8.5% by weight proline and about 2.5% -6.5% by weight serine and about 0% -1% by weight tyrosine and about 3% -8% by weight glycine and 3.5% -5.5% by weight glutamic acid and about 2% -4% by weight aspartic acid and large aspartic acid based on the total weight of said bioactive amino acids About 0% to about 1% by weight cysteine.

11. A method according to any one of claims 5 to 9, further comprising the addition of a nutritional supplement.

12. A method for preparing an aqueous solution for the treatment of atherosclerosis, characterized in that fatty substances are deposited on the inner layer of an arterial vessel and are fibrosed, resulting in plaque being deposited on the vessel wall and becoming denatured. The process comprises adding to the aqueous base an additive solution comprising an active amino acid selected from the group consisting of isoleucine, leucine, valine and arginine and mixtures thereof and adjusting the pH of the solution within the pH range of human blood, said additive solute being added to the solution in an amount effective to reduce plasma cholesterol levels, reverse plaque deposits and reverse the vascular degenerative conditions associated with atherosclerosis upon parenteral administration of said solution for a period of time effective.

13. The aqueous solution of claim 12 further comprising the step of adding lysine, methionine, phenylalanine, threonine, tryptophan and histidine to the aqueous solution.

14. The aqueous solution of claim 13, further comprising the step of adding alanine proline, serine, tyrosine, glycine, glutamic acid, aspartic acid and cysteine to the aqueous solution.

15. The aqueous solution of claim 12, 13 or 14 wherein the weight of said arginine added is the maximum weight percent of the total weight of the biologically active amino acids.

16. The method of any one of claims 12-15, wherein said biologically active amino acids comprise their levorotatory form.

17. The method of any one of claims 12-15, wherein said biologically active amino acids comprise a crystalline form thereof.

18. The method of any one of claims 12-17, wherein said solution comprises:

about 3% -10% by weight isoleucine and about 3% -10% by weight leucine and about 2.5% -7.5% by weight lysine and about 2.5% -7.5% by weight methionine and about 3% -10% by weight phenylalanine and about 1% -4% by weight threonine and about 1% -3% by weight tryptophan and about 3% -10% by weight valine and about 2% -20% by weight alanine and about 7% -20% by weight arginine and about 1% -4% by weight histidine and about 0% -10% by weight proline and about 0% -8% by weight serine and about 0% -1% by weight tyrosine and about 0% -10% by weight glycine and 0% -6% by weight glutamic acid and about 0% -6% by weight aspartic acid and large amino acids based on the total weight of said biologically active amino acids About 0% to about 1% by weight cysteine

19. The method of any one of claims 12-18, wherein said solution comprises:

about 4% -10% by weight isoleucine and about 5% -10% by weight leucine and about 4.5% -6.5% by weight lysine and about 3.5% -5.5% by weight methionine and about 4.5% -7.5% by weight phenylalanine and about 2% -3.5% by weight threonine and about 1% -2.5% by weight tryptophan and about 4% -9% by weight valine and about 7% -16% by weight alanine and about 9% -18% by weight arginine and about 1.5% -3.5% by weight histidine and about 4.5% -8.5% by weight proline and about 2.5% -6.5% by weight serine and about 0% -1% by weight tyrosine and about 3% -8% by weight glycine and about 3.5% -5.5% by weight glutamic acid and about 2% -4% by weight aspartic acid and about 4% by weight glutamic acid About 0% to about 1% by weight cysteine.

20. The method of any one of claims 12-19, further comprising the step of adding a nutritional supplement to the solutes.

21. The method of any of claims 12-20, wherein said solvent comprises a sterile deionized water base.

22. A method according to any one of claims 12-21, wherein said solute additive is present in said aqueous medium in a concentration of at least 4 grams, preferably 7.5-12 grams, and most preferably 9-12 grams of said biologically active amino acid per 100 milliliters of solution.