NO882017L - PROCEDURE AND SUBSTRATE MIXTURE FOR TREATMENT OF ATHEROSCLEROSIS. - Google Patents

Description

The present invention generally relates to compositions and methods for the treatment of atherosclerosis; more particularly, it relates to methods and substrate compositions for the treatment of atherosclerosis, whereby the many and varied problems associated with the disorder can be halted, greatly alleviated, or even partially cured. Even more particularly, the present invention utilizes mixtures of biologically active levorotatory (L-form) amino acids to achieve these results.

In the United States and Western Europe, heart/vascular disease and its associated diseases, dysfunctions, and complications are a significant cause of disability and the leading cause of death. One specific factor that significantly contributes to this pathophysiological process is atherosclerosis, which is generally recognized as the leading health care problem both in terms of mortality and health care costs. During each of the past five years, more than 550,000 deaths have occurred annually in the United States alone as a result of coronary artery disease. In addition, more than 680,000 hospitalizations occur annually for myocardial infarction, the biggest complication of atherosclerosis, and the number is constantly increasing. In recent years, more people in the United States have died as a result of atherosclerosis than from cancer, injuries and infectious diseases combined.

In 1983, direct health care costs for the care of patients with coronary heart disease in the United States exceeded $8 billion. During the same period, the total economic cost to this nation as a result of coronary heart disease as a manifestation of atherosclerosis alone was in excess of $60 billion and is estimated for 1987 to reach $96 billion. These costs do not even reflect the costs required for the care or treatment of the many and varied non-cardiac conditions caused by atherosclerosis.

Atherosclerosis is a disorder characterized by the deposition of fatty substances, mainly cholesterol, and subsequent fibrosis in the inner layer (intima) of an artery, resulting in plaque deposition on the surface of, and degenerative changes in, an arterial wall. The ubiquitous fatty plaque in the artery is the earliest lesion of atherosclerosis, often found even in children, and is a densely flattened, lipid-rich atheroma composed of both macrophages (white blood cells) and some smooth muscle fibers. The fibrous plaques representative of the different forms of advanced atherosclerosis have increased originally smooth muscle cells surrounded by a connective tissue matrix and contain variable amounts of intracellular and extracellular lipid. On the cavity surface of the artery, this plaque or lesion is usually covered by a dense fibrous capsule of smooth muscle or connective tissue. Under the fibrous capsule, the damage is largely to the cells consisting of macrophages, other leukocytes and smooth muscle cells. Deep in this cell-rich area, there may be areas of cholesterol crystals, necrotic debris and calcification.

If the disorder is allowed to progress, it can cause narrowing and occlusion of the cavity in the artery resulting in reduced or prevented blood flow and thus subsequent ischemia or infarction in the mainly affected organ or anatomical part such as the brain, heart, viscera or extremities. The result can be serious loss of function, loss of cell mass, medical and/or surgical emergency measures and significant disability or death. Alternatively, the cell wall can be severely weakened by infiltration into the muscle layer with the lipid (cholesterol), inflammatory white blood cells, connective tissue and calcium, resulting in soft and/or brittle areas that may in some areas be enlarged (aneurysm) and rupture or rupture , resulting in organ, limb or even life-threatening bleeding.

While the basic ultimate cause or causes of atherosclerosis are not fully known or understood, it has been hypothesized that this disorder is associated with plasma cholesterol and lipid levels based on empirical clinical observations by many researchers and medical practitioners. Therefore, one of the currently preferred regimens for the treatment of atherosclerosis consists of medication, diet, lifestyle modification, and exercise aimed at controlling and reducing plasma cholesterol levels.

Common medication used to lower plasma cholesterol levels include "Atromide-S" (clofibrate), "Choloxin" (dextrothyroxine sodium), "Colestid" (colestipol hydrochloride), "Lopid"

(gemfibrozil), "Lorelco" (probucol), "Nicolar" (niacin/nicotinic acid) and "Questran" (cholestyramine resin). However, these drugs and treatment in general target only the cause and not the result of atherosclerosis and have not been shown to be effective

when it comes to driving back the plaque deposit and degenerative changes in the arterial walls. The pharmacological agents also have many other faults, such as e.g. harmful side effects (hypertension, cardiac arrhythmias, stomach/intestinal disorders, headaches, hypersensitivity, etc.), contraindications (heart, liver or kidney disorders, pregnancy, etc.), requirements for lifelong careful administration, difficulties in maintaining patient cooperation, variable reliability and high price.

Once the disorder has progressed to the stage of severe persistent symptoms and compromised mobility, the next step of treatment has conventionally been "bypass" artery grafting to repair and/or replace the damaged artery. While coronary artery "bypass" has become one of the more common major cardiovascular surgeries in the United States, surgery is clearly not the solution to the pathological process as it has no arresting or reversing effect on the progression of the disorder and only temporarily overcomes the most critically attacked artery or arteries by bypassing it, if possible. Moreover, there is a significant risk of morbidity or mortality associated with surgery that many patients are unwilling to accept. Indeed, the disorder may continue to develop even while surgery is being performed, and the autogenous veins or arteries used to bypass the diseased arteries generally undergo atherosclerotic changes postoperatively at a faster rate than the originally affected arteries. Moreover, the conclusion reached by "the Coronary-ARTery Surgery Study (CASS)" supported by "the national Heart, Lung and Blood Institute (NHLBI)" is that there is no overall statistical improvement in the survival rate or the rate of development of myocardial infarction between the surgically and medically treated patient groups.

As an alternative to coronary bypass11 surgery, it has been suggested that certain medical treatments and procedures can be used to treat the results of atherosclerosis and not just the cause. These treatments include gelling ethylenediaminetetraacetic acid (EDTA) and percutaneous transluminal coronary angioplasty (PTCA ).EDTA treatments, however, are still experimental, unproven, and possibly as harmful as they are beneficial. PTCA treatments are invasive, of limited use and success, and occasionally result in fatal complications. Highly experimental intra -arterial laser beam plaque vaporization has limited application and requires an open operative approach to affected vessels.

It has now surprisingly been discovered that parenterally administered solutions comprising certain mixtures of biologically active amino acids are effective both in markedly reducing plasma cholesterol levels and in arresting and reversing arterial plaque deposition and vascular degenerative changes associated with atherosclerosis.

Certain amino acid solutions have been parenterally administered intravenously over the past three or four decades for a variety of medical reasons. Total parenteral nutrition (TPN), or intravenous hypernutrition, is now a widely accepted technique for the treatment of nutritional and metabolic problems in patients whose absorption and assimilation of sufficient optimal nutrient substrates cannot or should not take place via the alimentary canal. Indeed, the use of this technique, pioneered by the present inventor, has been successful in supporting normal growth and development in infants and children for more than fifteen years after birth.

Currently, several standard TPN solutions are commercially available in the United States, including those sold under the trade name "Travosol" (Travenol Laboratories), "Freamine"

(McGaw Laboratories), "Aminosyn" (Abbott Laboratories) and "Novamine" (Cutter Medical, division of Miles Laboratories). These standard intravenous nutrient solutions are designed to

to be nutritionally balanced, i.e. they are prepared to meet all the calorie, protein (amino acid), vitamin, mineral, water and trace element needs of the average patient. The components of these solutions are also available in modular form, enabling the physician and pharmacist to prepare special solutions tailored to the patients' specific needs during their treatment. Essential fatty acids and additional calories in the form of a lipid can also be administered separately intravenously as fortified soybean or safflower oil emulsions to meet specific nutritional needs.

For patients with kidney failure, liver failure or severe stress, special intravenous preparations of amino acids are also available for infusion as a supplement to the treatment of these conditions. E.g. US Patent No. 3,832,465 describes an amino acid solution for nutritional use, and US Patent No. 3,950,529 describes the administration of amino acids to patients who have liver disorders.

No suggestion has been made, however, that parenterally administered biologically active amino acid solutions could be used to treat atherosclerosis, particularly with a view to reversing the harmful complications associated with the disorder.

It is therefore an object of the present invention

to provide amino acid compositions and methods for treating atherosclerosis, whereby not only plasma cholesterol levels (contributing cause) are lowered, but also plaque deposition and degenerative arterial changes (results) and secondary complications associated with the disorder can be largely halted, reversed or even to a certain extent is cured.

It is also an object of the present invention

to provide amino acid mixtures and methods for the treatment of atherosclerosis which do not produce the many side effects of common drug treatments and can effectively reduce or even eliminate the need for major surgery or other invasive procedures in most atherosclerotic patients.

Summary of the invention

In accordance with the present invention, there are provided methods and substrate compositions for use in the treatment of atherosclerosis, a disorder characterized by the deposition of fatty substances in, and fibrosis of, the inner layers of an artery resulting in plaque deposition on, and degenerative changes in , an artery wall. The methods and compositions of the present invention are particularly advantageous in that they act both to significantly lower plasma cholesterol levels and to a large degree arrest, reverse and, to some extent, even cure arterial plaque deposition and degenerative vessel wall changes associated with atherosclerosis.

The mixtures according to the present invention comprise,

in its overall concept, at least in part an active antiatherosclerotic agent which in turn comprises a mixture of biological

active (L-form) amino acids. This mixture more preferably comprises at least arginine and the branched chain amino acids, isoleucine, leucine and valine, with arginine preferably present in the mixture in the largest percentage by weight based on the total weight of the amino acids in the mixture. Even more preferably, the mixture also comprises lysine, methionine, phenylalanine, threonine, tryptophan and histidine. Most preferably, the mixture also comprises alanine, proline, serine, tyrosine, glycine, glutamic acid, aspartic acid and cysteine.

The mixtures according to the present invention may also include, in addition to the amino acid mixture, other nutrients or additives such as vitamins, minerals, electrolytes, carbohydrates, antibiotics, anticoagulants and the like. as required by the specific needs of a patient.

The compositions of the present invention are preferably administered parenterally in the form of an aqueous solution through a central venous catheter due to the hypertonic nature of the preferred aqueous solutions. The daily ration of the aqueous solution is further administered preferably continuously and constantly intravenously with an intravenous infusion pump.

The main advantages of this invention over other methods of treating patients with atherosclerosis include the following: plasma cholesterol levels can be lowered more quickly and to a greater extent than previously achieved with any previous therapeutic agent; the degree and extent of atherosclerotic plaque clearance is greater than ever seen before; the duration of therapy required to drive back atherosclerosis plaque is relatively short compared to the duration of time over which the plaque originally developed; all dissolution ingredients are biologically active nutrient substrates with negligible toxicity or adverse side effects when administered within the suggested dosage range; the cost/effectiveness and cost/benefit ratios are favorable compared to pharmacological and surgical approaches to treating atherosclerosis; the therapy can be started and carried out exclusively on an outpatient basis; and the method addresses and attacks the basic pathophysiology of atherosclerosis primarily and biochemically rather than pharmacologically or by treating only the complications of atherosclerosis surgically.

These and other features and advantages of the present invention will be more easily understood by those skilled in the art by reading the following detailed description and examples with reference to the attached drawing.

Brief description of the drawing

Fig. 1 is a graphical representation of the daily total plasma cholesterol levels in a human patient, whose treatment is described in example 2, showing the response obtained by changes in the composition of the infusate.

Detailed description of the preferred design

The present invention provides methods and substrate mixtures for the treatment of atherosclerosis, which as previously mentioned is a disorder characterized by the deposition of fatty substances, mainly cholesterol, as plaque and accompanying fibrosis in the inner layer (intima) of an artery. As also previously mentioned, atherosclerosis is believed to be at least partially associated with plasma cholesterol and lipid levels. The compositions and methods of the present invention are particularly effective in both lowering plasma cholesterol levels (contributing cause) and also in reversing arterial plaque deposition and degenerative changes (result) to almost completely stop, alleviate and, to some extent, even cure the many and varied problems, conditions and secondary complications associated with the disorder.

The mixtures according to the present invention, in their overall design, consist of biological substrates that include,

at least in part, an active antiatherosclerotic agent. Antiatherosclerosis, used as here, refers to both

lowering plasma cholesterol levels and stopping and reversing arterial plaque deposition and degenerative changes in the artery wall. The active antiatherosclerotic agent, in the preferred embodiment, is a mixture of biologically active amino acids, preferably L-form amino acids.

Biologically active amino acids are generally classified as either essential or non-essential. Essential amino acids are those that cannot be synthesized in the body and must be consumed in sufficient quantities in the diet. Non-essential amino acids, on the other hand, are those that can be synthesized biochemically in the body from other substrate sources. The amino acids most commonly classified as essential include isoleucine, leucine, lysine, methionine, phenylalanine, threonine, tryptophan and valine. The amino acids most commonly classified as non-essential include alanine, arginine, histidine, proline, serine, tyrosine, glycine, glutamic acid, aspartic acid and cysteine. Arginine and histidine are classified as semi-essential in that they appear to be essential for young children and to a neon degree in the presence of renal failure.

These biologically active amino acids also exist in dextrorotatory or D-form and levorotatory or L-form. The L-form is considerably more biologically active than the D-form and is therefore preferred for use as an antiatherosclerotic agent.

The substrate mixtures according to the present invention are in an aqueous solution which in turn is administered parenterally to a patient as treatment. Thus administered, the compositions according to the present invention result in substantial lowering of plasma cholesterol levels and substantial repulsion of arterial plaque deposition on, and degenerative changes in, the arterial system.

The antiatherosclerotic agent is preferably a mixture of at least arginine, isoleucine, leucine and valine, preferably with arginine present in the largest percentage by weight based on the total weight of the biologically active amino acids in the mixture. These amino acids preferably make up at least 25 percent by weight of the amino acids present, but can be the only active ingredients for short infusion periods. The mixture may also include various of the other amino acids. Most preferably, all of the above listed amino acids are present in the composition to provide administration of all the amino acids together with other ingredients in a nutritionally balanced solution. All of these ingredients are part of TPN solutions, but surprisingly, increased levels of the branched chain amino acids and arginine, often associated with a reduction in the amount of histidine in the amino acid mixture, results in rapid (usually around 1 week) dramatic (usually 40-60 percent ) reduction in plasma cholesterol. Also noteworthy is the reduction of the simplest amino acids, especially alanine and glycine, especially glycine, from TPN solutions.

The amino acids are included in the preferred amino acid mixtures in the approximate weight percent ranges based on the total weight of the amino acids in the mixture, as shown in Table I below.

Note that the non-essential amino acids listed above which are possible ingredients in the broad sense of this invention may contribute less to atherosclerosis improvement than the other species. The same may be true for alanine and essential non-branched chain amino acids. However, these amino acids are important from a nutritional point of view when patient treatment extends over a period of months. As such, these amino acids can be added in larger or smaller amounts than stated above without significantly affecting or changing the characteristics and benefits of the solution and, therefore, without deviating from the application of the present invention. Thus, a more preferred amino acid mixture consisted of the foregoing amino acids present in the following approximate weight percentages based on the total weight of the amino acids in the mixture as set forth in Table II below.

The amino acid mixtures are preferably prepared by mixing the crystalline forms of the amino acids, usually the acetate, the chloride and the hydrochloride salt. A balancing of the ions used in the salts is necessary to avoid disturbing the acid/base balance in the patient's body; usually approx. 80 percent acetate and 20 percent chloride and hydrochloride. Usually, the pH in the solution according to the invention will vary from approx. 5.0 to approx. 7.8, but will preferably be adjusted to the blood's pH, approx. 7.39, by adding a suitable non-toxic organic compound. The crystalline form of amino acid is preferred because of its high purity and the precise, reproducible products that can be made therefrom. Furthermore, solutions made from the crystalline form can be directly assimilated into the body without prior biological impact in the body. The mixture of amino acids can also be prepared using commercially available parenteral nutrition (TPN) solutions and the addition of amino acids, preferably in crystalline form, to obtain solutions according to the invention. Preferred TPN solutions include those sold under the trademark "Travasol".

In addition, the mixtures according to the present invention can include other essential nutrients such as vitamins, minerals, electrolytes, carbohydrates, antibiotics, anticoagulants and the like as needed for nutritional, antibacterial and anti-caking purposes and other biological aids that will be known to someone who has professional knowledge. However, as with virtually all biological preparations, the most preferred mixtures will be patient-specific, that is, the relevant amino acid mixture can be varied based on a number of factors that normally determine drug mixtures and dosages such as, for example, the reaction of the particular patient to the mixture ; the specific patient's tolerance to the individual and combined components; the stage and degree of the disorder; the main organ(s) or system(s) adversely affected by the disorder; the primary biological functions adversely affected by the disorder and/or its complications; the occurrence, nature and degree of accompanying malnutrition, the patient's overall clinical symptom complex; the healing stage and from surgical and other methodological attempts to alleviate the signs and symptoms of the disorder and/or its complications; age, sex and general condition of the patient; presence of other accompanying disorders (diabetes mellitus, familial hyperlipidemia, etc.); and other factors that will be understood by those skilled in the art.

These factors are all related to a patient's ability to utilize or tolerate the amino acid composition and other components of the compositions of the present invention. In other words

each patient may require some adaptation of specific ingredients at different stages during treatment as a result of one or more of the above-mentioned factors. E.g. can one

diabetics may not be able to tolerate as high a calorie content in the form of sugar as a person without diabetes. As another example, it is known that the amino acid requirements of a person with kidney or liver failure are different from those of a person without these conditions. As yet another example, a person suffering from malnutrition may require greater amounts of, and different, amino acids and other nutrients than a well-nourished person. One skilled in the art will be able to recognize these particular needs and be able to adjust the compositions of the present invention accordingly.

Other adjustments to the amino acid mix can be determined from amino acid profiles in blood and urine. These profiles are signs of how the body utilizes a specific amino acid, and any deficiencies or excesses can be determined from such profiles and compensated for by changing the amino acid mixture. One skilled in the art will be able to make the desired adjustments accordingly once such an amino acid profile is determined. The amino acid profile is adjusted in response to blood and urine analysis for

to determine the utilization of amino acids by the patient's body. It has been found that the antiatherosclerotic effect is largely determined by the amount of branched chain amino acids and arginine in the treatment. The importance of arginine as an important amino acid has hitherto been overlooked, but, surprisingly, in the application of this invention emerged as an important contributor.

The mixtures according to the present invention are preferably administered parenterally in the form of an aqueous intravenous solution. The aqueous solution therefore comprises the mixture as a dissolved additive in solution with an aqueous solvent base. The aqueous solvent base preferably comprises a sterile, ion-free water base to which can be added a number of different components depending on the composition of the amino acid mixture and the particular needs of the individual patient. For example, if the mixture does not have sufficient caloric value and the patient needs additional caloric intake, the aqueous solvent base may be dextrose-water, fructose-water, or invert sugar-water solution (equal parts of dextrose and fructose) including variable concentrations of carbohydrate sub - the strategies that are chosen to fulfill the caloric needs of the patient. The solutions are mixed using established aseptic, antiseptic and pharmacochemical techniques well known to those with specialist knowledge to ensure the complete solubility of the ingredients while maintaining stability, sterility, safety and the preferred preparation for the individual patients.

It is preferred that the active antiatherosclerotic agent, i.e., the amino acid mixture, is present in solution in an effective amount which, when the solution is administered parenterally using hyperalimentation techniques well known to those skilled in the art, results in substantial lowering of plasma cholesterol and substantial repulsion of arterial plaque - deposition and degenerative artery wall changes after a relatively short administration period, usually within 2-6 months, while at the same time providing the patient's body with the nutrition necessary to sustain life. The amino acid mixture is present in the aqueous solution in concentrations varying from as little as approx. 4

g total weight of amino acids per 100 ml solution (a 4% solution by weight) up to the solubility limits of the specific amino acid mixture in the particular aqueous solvent (sometimes as high as a 20% solution by weight). Preferably, the amino acid mixture is present in solution in concentrations varying from about 7.5 to approx. 12 g total weight of amino acids per 100 ml of solution (a solution of 7.5 percent to 12 percent by weight). These concentrations will result in a hypertonic solution (800-1100 milliosmoles per liter), i.e. a solution that has a higher osmotic pressure than blood (290-310 milliosmoles per liter), the significance of which is discussed below. For the specific effective mixture described above, it is preferred that the amino acid mixture is present in solution at a concentration of from about 9 to approx. 11, more preferably from approx. 10 to approx. 11, g amino acids per 100 ml solution.

The aqueous solutions according to the present invention are administered parenterally, as the oral route has a number of disadvantages. Administered orally, the amino acids are exposed to the modifying digestive action of the stomach, intestinal tract, pancreas and liver; the endogenous cycle exerts a buffering effect on the components of the preparation; the pattern of solutions to be administered is difficult to determine so that the resulting blood levels of the active antiatherosclerotic agents are obtained identical to those effected by direct intravenous infusion; and procedures for controlling precise levels of intake are difficult to establish and maintain. With parenteral administration, a direct attack can be made against target tissue, and in some cases it is even possible to isolate a part of the body and perfuse it with amino acid solution for hours, days or weeks without drastically changing the patient's normal nutritional pattern. Furthermore, parenteral administration allows constant, controlled administration, thus ensuring a continuity of action without the troughs and peaks that inevitably occur after oral or enteral administration and accompanying absorption and assimilation.

Since the preferred aqueous solution is hypertonic (about 800 to about 1100 milliosmoles per liter), these solutions should be administered intravenously through a large-diameter blood vessel that has high blood flow for approximately instantaneous dilution, most preferably through a central venous catheters are inserted percutaneously into the superior vena cava by any of the various acceptable methods known to those skilled in the art. A particularly preferred method of catheter insertion is described in the article by O'Donnell, Calgue and Dudrick, entitled "Percutaneous Tunnel for Intravenous Hyperalimentation," published in the Southern Medical Journal, vol. 76, No. 11 (1983), pages 1344-48, which is hereby incorporated herein by reference for all purposes as if fully set forth.

After catheterization is achieved, continuous infusion of the aqueous antiatherosclerotic solution can begin. The total daily dissolution ration to the individual patient is preferably administered in an amount sufficient to provide the patient with between approx. 0.25 and approx. 3.5 g of amino acids, more preferably between approx. 1 and approx. 2 g amino acids, per kg body weight per 24 hour day. This solution ration is also preferably administered continuously and constantly intravenously by commercially available intravenous infusion pumps in order to maintain a homogeneous amino acid level in the body.

During the course of prolonged infusion therapy, generally from approx. 2 months to approx. 8 months, the patient preferably does not consume additional nutrient substrates that may affect the amino acid balance or metabolism in the body. The patient will be allowed to drink non-nutritive fluids and take any oral medication that may be necessary during the course of therapy. In selected patients, it will be possible to infuse the solution on an ambulatory or outpatient basis with the use of portable, lightweight pumps and a tailored carrier for the solution, infusion tube and pump. For even more selected patients, it may be preferable to infuse the total daily ration of solution over 12-16 hours of the 24-hour day, and allow patients to disconnect themselves from the pump and infusion equipment for 8-12 hours to increase their freedom and activities during the non-infusion periods. When the patient is disconnected from the infusion tubes, it will be necessary to drip in 1-2 ml of solution containing approx. 100 units of heparin per ml into the central catheter and close it with a sterile cap (e.g. "Luer-lock") to prevent clogging of the catheter with a blood clot.

During the first weeks of infusion of the antiatherosclerotic solution, the patient should be monitored biochemically, haematologically and metabolically every 1-3 days via blood and urine samples. Amino acid profiles from serum and/or urine can be taken from the samples as desired and the amino acid mixtures adjusted accordingly. Once initial stability of these signs has been achieved in response to the aqueous solution infusion and any indicated changes have been made to the initial composition of the solution, monitoring intervals may be extended to every 1-2 weeks.

Other signs that must be monitored include the following: serum cholesterol, cholesterol esters, apolipoprotein A^, apolipoprotein B100»high-density lipoprotein (HDL) cholesterol, triglycerides

there, sodium, potassium, chloride, bicarbonate, creatinine, calcium, inorganic phosphorus, glucose, urea nitrogen, uric acid, total protein, albumin, bilirubin, alkaline phosphatase, LDH, SGOT, SGPT,

GGT, CPK, magnesium, zinc copper, prothrombin time in blood, partial thromboplastin time, leukocyte count, erythrocyte count, hemoglobin,

hematocrit, mean blood cell volume, mean blood cell haemoglo-

bin, mean corpuscular hemoglobin concentration, differential leukocyte count and platelet count. These standard tests are used individually, in varying combinations and collectively to ensure the safety and effectiveness of the treatment, to maintain homeostasis and further assist the clinician in adjusting the solution specific to the individual patient.

The foregoing detailed discussion of this invention

will further be exemplified by the following specific examples

is given by way of illustration and not limitation of the invention described above.

Example 1

Attempts to implicate cholesterol as one of the major etiological agents in atherogenesis have been somewhat weakened by the inability to reduce and maintain lowered serum cholesterol levels by diet and/or chemotherapeutic agents alone.

In a series of preparatory studies, an animal model of atherosclerosis was developed over a period of several years by modifying the diet of New Zealand albino rabbits. Using several hundred male and female rabbits in various studies with successive and logical changes of the diets, a consistent and reliable animal model was developed for the development of atherogenesis in these rabbits that mimicked the distribution, different stages and severity, overall morphological and histological characteristics , primary pathophysiology, secondary complications and clinical manifestations of atherosclerosis corresponding to that observed and documented in humans.

Rabbits, as herbivores, usually consume only small amounts of dietary cholesterol. The inability of the rabbit livers of these rabbits to biochemically remove large amounts of dietary cholesterol resulted in rapid increases in plasma cholesterol concentration, which in turn resulted in a reproducible degree, pattern, and extent of atherosclerosis to advanced stages over a defined time period proportional to elevation of plasma cholesterol concentration and duration of hypercholesterolaemia.

As a result of the preliminary studies, an atherogenic diet was prepared and used to produce atherosclerosis in several hundred additional rabbits which were used as subjects in the following studies. In a typical study, 40 New Zealand albino adult male and female rabbits were all fed a basic laboratory rabbit diet ("Teklad" T.D. 82135) consisting of high fiber "Purina" Labchow 5326 supplemented with 8% by weight peanut oil as a carrier for 2% by weight crystalline cholesterol. Within 10 days to 2 weeks of starting this oral dietary regimen, total plasma cholesterol levels rose to approx. 1000 mg/dl and was held between 1000 and 2000 mg/dl for the remainder of the six-week atherogenic period.

After six weeks, ten rabbits were killed and autopsied to image and document the gross and microscopic severity, stage and extent of disease in these animals as a baseline. The remaining 30 rabbits in the original groups were randomly assigned to one of three study groups. Group I consisted of ten animals that continued the atherogenic diet regimen for an additional six weeks. Group II consisted of ten animals receiving normal laboratory rabbit diet ("Teklad" T.D. 82135) without peanut oil and crystalline cholesterol for six weeks. Group III consisted of ten animals that were continuously infused with an antiatherosclerotic solution for six weeks via a central venous catheter placed in the superior vena cava in a manner known to those skilled in the art. The amino acid composition of the antiatherosclerotic solution containing 4.25 percent (by weight) amino acids and 25 percent (by weight) dextrose is listed below in Table III (column A). The solution was administered to provide 1.5 g of amino acids and 35 calories per kg of body weight daily with vitamins, minerals and trace elements added to achieve a balanced nutritional mix.

At the end of the second six-week study period, all 30 rabbits were killed and necropsied to determine and image the severity, condition, and distribution of atherosclerosis. In group I rabbits, complicated atherosclerosis damage was inflicted as reproducible and uniform comprising approx. 85 percent to 95 percent of the aorta from its origin at the heart to the bifurcation of the common pelvic arteries, with a pattern of superficial plaque and arterial wall distribution characteristic of atherosclerotic lesions observed in humans. Several of the rabbits in group I showed signs of having undergone human-like myocardial infarction, cerebellar infarction, visceral ischemia and infarction, and hindlimb ischemia and infarction.

The rabbits in group II fed the standard laboratory rabbit diet for six weeks showed no regression of the atherosclerotic plaque compared to the first group of ten rabbits that were killed after six weeks on the atherogenic diet.

Instead, there was a slight development of atherosclerosis in group II

during this second six-week period.

However, in the group III rabbits that received the special atherosclerosis preparation intravenously, a dramatic decline in the atherosclerotic plaque occurred. In fact, the aorta was approx. 90 percent for some traces of atherosclerosis. In response to amino acid concentrations in plasma and urine measured at weekly intervals in the group III rabbits, the daily intravenous amino acid ration was increased from 1.5-1.7 g per kg body weight, and glutamic acid and aspartic acid were added to the preparation. The final amino acid composition for Group III rabbits is shown in column B, Table III. Note that the greatest increase occurred in the branched chain amino acids in the mixture, while arginine was increased only slightly. These experiments were repeated until a total of 200 rabbits had been examined in each of the three groups, with results consistent with those above.

This study shows consistent reduction of atherosclerotic plaque as a result of only intravenous amino acid treatment. These investigations form the basis and justification for the development of subsequent investigations in human patients.

Chronologically, the investigations at this time were started in human patients using the antiatherosclerotic solution of the same composition that was infused in the last group of forty rabbits.

Example 2

A 40 year old man with severe generalized atherosclerosis secondary to familial type II hyperlipidemia, previously treated at a lipid trial clinic with a combination of low cholesterol/fat, low calorie (1400 calorie) diet and cholestyramine was considered for treatment using solutions of this invention . Even after five months of this previous treatment, the patient's plasma cholesterol level could only be reduced from a baseline of 496 mg/dl to 409 mg/dl (upper normal plasma cholesterol level is 260 mg/dl), and the patient did not feel any improvement in his symptoms specified below. Within two weeks of voluntary discontinuation of this first regimen, plasma cholesterol returned to levels between 490 and 536 mg/dl.

The patient recovered from a myocardial infarction one year earlier, but continued to have angina pectoris, could only walk a quarter of an hour without severe leg cramps and claudication, and had frequent fainting spells and numbness in his extremities due to poor blood flow in the main arteries. Four of the eight distal extremity pulses that are usually felt in the arms and legs were absent, and the remaining ones were reduced in control by approx. 50 percent. Coronal angiography showed complete occlusion of his right main coronary artery with diffuse narrowing and poor blood flow in his left internal descending and surrounding coronary artery due to atherosclerosis. There is no evidence of significant parallel myocardial circulation, and because of the overall diffuse distribution of his disease, he was not a possible candidate for coronary artery "bypass" surgery.

His life prognosis was less than two years. His plasma cholesterol concentration was 540 mg/dl, he was barely mobile and fainting attacks had increased in frequency and strength, so that he had fallen and injured himself on several occasions. Magnetic resonance imaging of his abdominal aorta, pelvic arteries and carotid arteries confirmed up to 80 percent blockage of the cavities in these vessels by atherosclerosis plague. Doppler ultrasound examinations of both lower extremities showed markedly reduced arterial blood flow bilaterally.

A percutaneous central venous catheter was inserted into the superior vena cava via the right clavicle vein, and initially an intravenous solution of amino acid composition as shown in Column B, Table III above was continuously infused. The patient initially received 1.35 g of amino acids per kg body weight, or a ration of 95 g of amino acids such as this preparation. At the same time, he received 35 calories per kg body weight as dextrose together with their daily requirement of vitamins, minerals and trace elements using common intravenous additives. Baseline metabolic investigations were recorded continuously during the six-month treatment and clinical investigation period. The patient was asked not to consume food other than water during the entire examination.

During the course of antiatherosclerosis infusion therapy and examination of the patient, the original solution composition was modified on several occasions in response to plasma cholesterol levels as shown in fig. 1.

As can be seen in fig. 1, the plasma cholesterol concentration fell from the initial 508 mg/dl to 450 mg/dl within two days. At point 1, the total daily calories were reduced in the mixture from 2500 (35 cal/kg) to 2100 (30 cal/kg) because the relative inactivity of the patient did not justify the originally calculated caloric needs. By the eighth day of therapy, the plasma cholesterol level had fallen further to 370 mg/dl, but rose to 393 mg/dl on the ninth day. At point 2, the daily dose of branched-chain amino acids — leucine, isoleucine and valine — was increased by two grams. At this point, the following amino acids were present in the following approximate weight percentages based on the total weight of the amino acids in the mixtures: isoleucine, 7.9%; leucine, 8.9%; lysine, 5.9%; methionine, 5.0%; phenylalanine, 5.9%; threonine, 3.0%; tryptophan, 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%; and cysteine, 0.6%. The amino acid dose was increased from 1.35 to 1.45 g per kg body weight per day to a total daily ration of 101.5 g of amino acids as this preparation.

Within ten days of starting treatment, the plasma cholesterol level had dropped dramatically to 200 mg/dl, which represented a 57 percent reduction from the baseline level of 508 mg/dl. At points 3, 4 and 5, minor changes in the concentrations of various amino acid components and dextrose calories were made in response to biochemical determinations made in serum and urine samples. More specifically, at point 3, phenylalanine was increased by 1 g, and the glycine dose was decreased by 1 g in the daily amino acid ration. At point 4, the daily dextrose ratio was increased to give 35 cal/kg, because the patient had increased his activity and started to lose weight. At point 5, the alanine dose was reduced by 3 g, while the glutamic acid and aspartic acid doses were both increased by 1.5 g in the daily amino acid ration.

At point 6, a dramatic rise in plasma cholesterol levels from 260 to 370 mg/dl occurred after intravenous infusion of a single 500 mL dose of 10 percent soybean oil emulsion (sold under the trade name "Intralipid"). Points 7 and 8 represent reductions in total calories administered as dextrose to 34 cal/kg and 33 cal/kg, respectively, the plasma cholesterol level was significantly elevated above its trough level before fat infusion for at least four weeks after infusion of the fat emulsion. By that time it had fallen back to 300 mg/dl, and at point 9 a single 500 ml dose of 20 per cent soybean oil emulsion was given intravenously ("Intralipid") with an increase in the serum plasma cholesterol level to 345 mg/dl, confirming the deleterious effect on the plasma cholesterol level by infusion of even essential fatty acids such as soybean oil emulsions. At point 10, the daily dextrose calories were reduced to 30 cal/kg, and the daily doses of proline and serine were both reduced by 2 g. Because the patient noted a loss of energy and weight, glucose was restored to 35 cal/kg at point 11 At point 12, the arginine concentration almost doubled in the solution, while the histidine was reduced. A significant drop in plasma cholesterol occurred from 306 mg/dl to 245 mg/dl. The composition of the amino acid mixture at this point is set forth in Table IV below. The amino acid dose at this point was increased from 1.45 to 1.5 g per kg body weight per day to a daily total ration of 103.5 g amino acids.

Over the next week, coronary angiography, magnetic resonance imaging of his aorta, carotid and pelvic arteries, and Doppler ultrasound examinations of both lower extremities were repeated. The treatment/investigation period was then terminated and the patient resumed oral intake with a restriction in dietary cholesterol to 100 mg daily.

During the six months of treatment, the patient had no angina pectoris or fainting attacks; three of the previously four absent extremity pulses could now be felt; the four previously palpable extremity pulses increased in strength by 50-100 percent; the blood flow to his legs constantly increased up to 250-300 percent as shown by resting doppler ultrasound examinations and he was able to walk up to 4.8-6.4 km per hour. day at 6.4 km/h without cramp or claudication in his legs.

At the end of the six-month treatment period, magnetic resonance imaging showed up to a 40-50 percent reduction in atherosclerosis plaque in the abdominal aorta, common pelvic arteries and carotid arteries. Repeat coronary angiography showed widely patent left anterior descending immediate coronary arteries, with several subordinate arteries to the right coronary artery also widely patent, except at the point of previous complete occlusion at its origin.

In addition to these dramatic demonstrations of atherosclerotic plaque repulsion and increased arterial blood flow through a number of previously at-risk arteries, further evidence of improved bodily function was shown by the patient's ability to play 18 holes of golf with scoring in the 70s and gain employment as a carpenter at a house construction company. He has remained free of symptoms and is physically active for more than fifteen months after his treatment was started. Continued periodic observation and examination took place to document the successful achievement of repulsion of atherosclerosis and improvement of arterial blood flow and body functions, as clearly shown in this patient, hitherto unknown with any form of medical or dietary treatment.

The final composition of solution obtained during the six month study period with this patient was then infused into an additional ten rabbits as part of an additional group of forty rabbits examined in an otherwise identical manner as described above. Reversal of atherosclerosis shown in this additional group of ten rabbits was the most effective, uniform and complete of any of the animals examined and ranged from 98-99 percent disappearance of atherosclerotic disease.

Example 3

Further confirmatory evidence of the effectiveness of the method and composition in the treatment of atherosclerosis was obtained with 30 volunteer human subjects who had received total parenteral nutrition via central venous catheters for two months to eight years. Reduction of plasma cholesterol levels by 40-60 percent below their normal cholesterol levels regardless of the magnitude of the absolute values was achieved in all subjects within one week of changing the aqueous solution infusate to the amino acid mixture as shown in either Table III or Table IV. These findings are very significant not only because of the magnitude of the reductions in plasma cholesterol levels documented in these subjects, but also because the positive response value was 100 percent. All these patients had received long-term intravenous infusion of common TPN substrates containing mixtures of amino acids, without any significant reduction in plasma cholesterol levels. The infusate mixture from Table IV caused dramatic reduction.

A striking example of the effectiveness of the solution described in Table IV in reducing plasma cholesterol levels in one patient occurred with the reduction of an already low baseline plasma cholesterol concentration of 83 mg/dl to 50 mg/dl within one week, representing a 40 percent reduction in using an aqueous solution having an amino acid mixture as set forth in Table IV.

These data suggest that the mechanism of action of this invention is probably an active biochemical process, rather than the result of strict restriction of dietary cholesterol, cholesterol precursors and other lipids, and demonstrate the effectiveness of the present invention.

The mixtures and methods according to the present invention, as previously described and as illustrated in the examples, provide a unique, strong and effective aid for the treatment of atherosclerosis. The recently completed "National Institute of Health"-supported "Lipid Research Clinic-Coronary Primary Prevent Trial" was the first human study to definitively establish that lowering blood cholesterol levels reduces heart attack and heart attack death. However, despite intense dietary control and the administration of pharmacologic plasma cholesterol-lowering agents, the average plasma cholesterol level during the ten-year study period was reduced by only 8.6 percent with no more than a 19 percent reduction in the best responding patients. However, the incidence of myocardial infarction in this study group was reduced by up to 19 percent compared to the general population. The incidence of myocardial infarction may be even more substantially reduced in conjunction with the 40-60 percent reductions in plasma cholesterol levels demonstrated by use of this invention.

It should be noted that many modifications and variations besides the designs specifically mentioned can be made in the previously described mixtures and methods without deviating significantly from the meaning of the present invention. Therefore, it must be clearly understood that the form of the invention described and illustrated here is only by way of example, and is not intended as a limitation on its scope.

Claims (43)

1. Antiatherosclerotic agent for lowering plasma cholesterol levels and reversing arterial plaque deposition and degenerative changes associated with atherosclerosis, consisting mainly of a mixture of biologically active amino acids, said mixture comprising: from about 3% to about 10% isoleucine from about 3% to about 10% leucine from about 2.5% to about 7.5% lysine from about 2.5% to about 7.5% methionine from about 3% to about 10% phenylalanine from about 1% to about 4% threonine from about 1% to about 3% tryptophan from about 3% to about 10% valine from about 2% to about 20% alanine from about 7% to about 20% arginine from about 1% to about 4% histidine from about 0% to about 10% proline from about 0% to about 8% serine from about 0% to about 1% tyrosine from about 0% to about 10% glycine from about 0% to about 6% glutamic acid from about 0% to about 6% aspartic acid from about 0% to about 1% cysteine by weight based on the total weight of said biologically active amino acids in said mixture. 2. Antiatherosclerotic agent according to claim 1, characterized in that said mixture further comprises: from about 4% to about 10% isoleucine from about 5% to about 10% leucine from about 4.5% to about 6.5% lysine from about 3.5% to about 5.5% methionine from about 4.5% to about 7.5% phenylalanine from about 2% to about 3.5% threonine from about 1% to about 2.5% tryptophan from about 4% to about 9% valine from approximately 7% to approximately 16% alanine from about 9% to about 18% arginine from about 1.5% to about 3.5% histidine from about 4.5% to about 8.5% proline from about 2.5% to about 6.5% serine from about 0% to about 1% tyrosine from about 3% to about 8% glycine from about 3.5 to about 5.5% glutamic acid from about 2% to about 4% aspartic acid from about 0% to about 1% cysteine by weight based on the total weight of said biologically active amino acids in said mixture. 3. Antiatherosclerotic agent according to claim 1, characterized in that arginine is present in said mixture in the largest percentage by weight based on the total weight of said biologically active amino acids in said mixture. 4. Antiatherosclerotic agent according to claim 1, characterized in that said biologically active amino acids consist of their left-handed form. 5. Antiatherosclerotic agent according to claim 1, characterized in that said biologically active amino acids consist of their crystalline form. 6. Mixture useful in the treatment of atherosclerosis, characterized by an active antiatherosclerotic mixture consisting mainly of a mixture of biologically active amino acids comprising at least isoleucine, leucine, valine and arginine present in said mixture in an effective amount, whereby parenteral administration of said mixture in form of an aqueous solution for an effective period of time results in a substantial lowering of plasma cholesterol and the substantial repulsion of arterial plaque deposition and degenerative changes associated with atherosclerosis and whereby arginine is present in said mixture in the largest percentage by weight based on the total weight of said biologically active amino acids in said mixture. 7. Mixture according to claim 6, characterized in that said mixture further comprises lysine, methionine, phenylalanine, threonine, tryptophan and histidine. 8. Mixture according to claim 7, characterized in that said mixture further comprises alanine, proline, serine, tyrosine, glycine, glutamic acid, aspartic acid and cysteine. 9. Mixture according to claim 6, characterized in that said biologically active amino acids consist of their left-handed form. 10. Mixture according to claim 6, characterized in that said biologically active amino acids consist of their crystalline form. 11. Mixture according to claim 6, characterized in that said mixture further comprises the following mixture of amino acids: from about 3.5% to about 10% isoleucine from about 3% to about 10% leucine from about 2.5% to about 7.5% lysine from about 2.5% to about 7.5% methionine from about 3% to about 10% phenylalanine from about 1% to about 4% threonine from about 1% to about 10% tryptophan from about 3% to about 10% valine from about 2% to about 20% alanine from about 7% to about 20% arginine from about 1% to about 4% histidine from about 0% to about 10% proline from about 0% to about 8% serine from about 0% to about 1% tyrosine from about 0% to about 10% glycine from about 0% to about 6% glutamic acid from about 0% to about 6% aspartic acid from about 0% to about 1% cysteine by weight based on the total weight of said biologically active amino acids in said mixture. 12. Mixture according to claim 11, characterized in that said mixture further comprises: from about 4% to about 10% isoleucine from about 5% to about 10% leucine from about 4.5% to about 6.5% lysine from about 3.5% to about 5.5% methionine from about 4.5% to about 7.5% phenylalanine from about 2% to about 3.5% threonine from about 1% to about 2.5% tryptophan from about 4% to about 9% valine from approximately 7% to approximately 16% alanine from about 9% to about 18% arginine from about 1.5% to about 3.5% histidine from about 4.5% to about 8.5% proline from about 2.5% to about 6.5% serine from about 0% to about 1% tyrosine from about 3% to about 8% glycine from about 3.5% to about 5.5% glutamic acid from about 2% to about 4% aspartic acid from about 0% to about 1% cysteine by weight based on the total weight of said biologically active amino acids in said mixture. 13. Mixture according to claim 6, further characterized in that it comprises a nutritional supplement. 14. Aqueous solution for use in the treatment of atherosclerosis, a disorder characterized by the deposition of fatty substances in, and fibrosis of, the innermost layer of an artery resulting in plaque deposition on and degenerative changes in an arterial wall, characterized in that it consists of: an aqueous base; and an added solute comprising an active antiatherosclerotic agent consisting mainly of a mixture of biologically active amino acids comprising isoleucine, leucine, valine and arginine, said added solute being present in said solution in an effective amount, whereby parenteral administration of said solution in an effective period results in a substantial lowering of plasma cholesterol and the substantial repulsion of arterial plaque deposition and degenerative changes associated with atherosclerosis. 15. Aqueous solution according to claim 14, characterized in that said mixture further comprises lysine, methionine, phenylalanine, threonine, tryptophan and histidine. 16. Aqueous solution according to claim 15, characterized in that said mixture further comprises alanine, proline, serine, tyrosine, glycine, glutamic acid, aspartic acid and cysteine. 17. Aqueous solution according to claim 14, characterized in that arginine is present in said mixture in the largest percentage by weight based on the total weight of said biologically active amino acids in said mixture. 18. Aqueous solution according to claim 14, characterized in that said biologically active amino acids consist of their levorotatory form. 19. Aqueous solution according to claim 14, characterized in that said biologically active amino acids consist of their crystalline form. 20. Aqueous solution according to claim 14, characterized in that said mixture further comprises: from about 3% to about 10% isoleucine from about 3% to about 10% leucine from about 2.5% to about 7.5% lysine from about 2.5% to about 7.5% methionine from about 3% to about 10% phenylalanine from about 1% to about 4% threonine from about 1% to about 3% tryptophan from about 3% to about 10% valine from about 2% to about 20% alanine from about 7% to about 20% arginine from about 1% to about 4% histidine from about 0% to about 10% proline from about 0% to about 8% serine from about 0% to about 1% tyrosine from about 0% to about 10% glycine from about 0% to about 6% glutamic acid from about 0% to about 6% aspartic acid from about 0% to about 0% cysteine by weight based on the total weight of said biologically active amino acids in said mixture. 21. Aqueous solution according to claim 20, characterized in that said mixture further comprises: from about 4% to about 10% isoleucine from about 5% to about 10% leucine from about 4.5% to about 6.5% lysine from about 3.5% to about 5.5% methionine from about 4.5% to about 7.5% phenylalanine from about 2% to about 3.5% threonine from about 1% to about 2.5% tryptophan from about 4% to about 9% valine from about 7% to about 16% alanine from about 9 % to about 18% arginine from about 1.5% to about 3.5% histidine from about 4.5% to about 8.5% proline from about 2.5% to about 6.5% serine from about 0% to approximately 1% tyrosine from approximately 3% to approximately 8% glycine from approximately 3.5% to approximately 5.5% glutamic acid from approximately 2% to approximately 4% aspartic acid from about 0% to about 1% cysteine by weight based on the total weight of said biologically active amino acids in said mixture. 22. Aqueous solution according to claim 14, characterized in that said added dissolved substance further comprises a nutritional supplement. 23. Aqueous solution according to claim 14, characterized in that said solvent comprises a sterile ion-free water base. 24. Aqueous solution according to claim 14, characterized in that said added dissolved substance is added to said aqueous base in amounts which give a concentration of at least approx. 4 g of said biologically active amino acids per 100 ml solution. 25. Aqueous solution according to claim 24, characterized in that said added dissolved substance is added to said aqueous base in amounts that give a concentration from approx. 7.5 to approx. 12 g of said biologically active amino acids per 100 ml solution. 26. Aqueous solution according to claim 25, characterized in that said added dissolved substance is added to said aqueous base in amounts that give a concentration that has from approx. 9 to approx. 11 g of said biologically active amino acids per 100 ml solution. 27. Method for treating atherosclerosis, a disorder characterized by deposition of fatty substances in and fibrosis of the innermost layer of an artery resulting in plaque deposition on and degenerative changes in an arterial wall, comprising parenteral administration for an effective period of time of an effective dose of an aqueous solution comprising an antiatherosclerotic agent which substantially lowers plasma cholesterol levels and substantially reverses said plaque deposition and said degenerative changes, characterized in that said antiatherosclerotic agent comprises a mixture of biologically active amino acids. 28. Method according to claim 27, characterized in that said mixture comprises at least arginine, isoleucine, leucine and valine. 29. Method according to claim 28, characterized in that said mixture further comprises lysine, methionine, phenylalanine, threonine, tryptophan and histidine. 30. Method according to claim 29, characterized in that said mixture further comprises alanine, proline, serine, tyrosine, glycine, glutamic acid, aparagic acid and cysteine. 31. Method according to claim 27, characterized in that arginine is present in said mixture in the largest percentage by weight based on the total weight of said biologically active amino acids in said mixture. 32. Method according to claim 27, characterized in that said biologically active amino acids consist of their left-handed form. 33. Method according to claim 27, characterized in that said biologically active amino acids consist of their crystalline form. 34. Method according to claim 27, characterized in that said mixture further comprises: from about 3% to about 10% isoleucine from about 3% to about 10% leucine from about 2.5% to about 7.5% lysine from about 2.5% to about 7.5% methionine from about 3% to about 10% phenylalanine from about 1% to about 4% threonine from about 1% to about 3% tryptophan from about 3% to about 10% valine from about 2% to about 20% alanine from about 7% to about 20% arginine from about 1% to about 4% histidine from about 0% to about 10% proline from about 0% to about 8% serine from about 0% to about 1% tyrosine from about 0% to approximately 10% glycine from approximately 0% to approximately 6% glutamic acid from approximately 0% to approximately 6% aspartic acid from about 0% to about 1% cysteine by weight based on the total weight of said biologically active amino acids in said mixture. 35. Method according to claim 34, characterized in that said mixture further comprises: from about 4% to about 10% isoleucine from about 5% to about 10% leucine from about 4.5% to about 6.5% lysine from about 3.5% to about 5.5% methionine from about 4.5% to about 7.5% phenylalanine from about 2% to about 3.5% threonine from about 1% to about 2.5% tryptophan from about 4% to about 9% valine from about 7% to about 16% alanine from about 9 % to about 18% arginine from about 1.5% to about 3.5% histidine from about 4.5% to about 8.5% proline from about 2.5% to about 6.5% serine from about 0% to approximately 1% tyrosine from approximately 3% to approximately 8% glycine from approximately 3.5% to approximately 5.5% glutamic acid from approximately 2% to approximately 4% aspartic acid from about 0% to about 1% cysteine by weight based on the total weight of said biologically active amino acids in said mixture. 36. Method according to claim 27, characterized in that said aqueous solution further comprises an aqueous base. 37. Method according to claim 36, characterized in that said aqueous base comprises a sterile, ion-free water base. 38. Method according to claim 36, characterized in that said aqueous solution further comprises a calorie supplement. 39. Method according to claim 27, characterized in that said antiatherosclerotic agent is added to said aqueous base in amounts that give a concentration of at least approx. 4 g of said biologically active amino acids per 100 ml solution. 40. Method according to claim 39, characterized in that said antiatherosclerotic agent is added to said aqueous base in amounts that give a concentration from approx. 7.5 to approx. 12 g of said biologically active amino acids per 100 ml solution. 41. Method according to claim 40, characterized in that said antiatherosclerotic agent is added to said aqueous base in amounts to give a concentration from approx. 9 to approx. 11 g of said biologically active amino acids per 100 ml solution. 42. Method according to claim 27, characterized in that said aqueous solution is administered parenterally in an effective dose consisting of from approx. 0.25 to approx. 3 total g of biologically active amino acids per kg body weight per day. 43. Method according to claim 42, characterized in that said aqueous solution is administered parenterally in an effective dose comprising from approx. 1 to approx. 2 total g of biologically active amino acids per kg body weight per day. CHANGED REQUIREMENTS: 1. Antiatherosclerotic agent for lowering plasma cholesterol levels and reversing arterial plaque deposition and degenerative changes associated with atherosclerosis, consisting mainly of a mixture of biologically active amino acids, said mixture comprising: from about 3% to about 10% isoleucine from about 3% to about 10% leucine from about 2.5% to about 7.5% lysine from about 2.5% to about 7.5% methionine from about 3% to about 10% phenylalanine from about 1% to about 4% threonine from about 1% to about 3% tryptophan from about 3% to about 10% valine from about 2% to about 20% alanine from about 7% to about 20% arginine from about 1% to about 4% histidine from about 0% to about 10% proline from about 0% to about 8% serine from about 0% to about 1% tyrosine from about 0% to about 10% glycine from about 0% to about 6% glutamic acid from about 0% to about 6% aspartic acid from about 0% to about 1% cysteine by weight based on the total weight of said biologically active amino acids in said mixture. 2. Antiatherosclerotic agent according to claim 1, characterized in that said mixture further comprises: from about 4% to about 10% isoleucine from about 5% to about 10% leucine from about 4.5% to about 6.5% lysine from about 3.5% to about 5.5% methionine from about 4.5% to about 7.5% phenylalanine from about 2% to about 3.5% threonine from about 1% to about 2.5% tryptophan from about 4% to about 9% valine from approximately 7% to approximately 16% alanine from about 9% to about 18% arginine from about 1.5% to about 3.5% histidine from about 4.5% to about 8.5% proline from about 2.5% to about 6.5% serine from about 0% to about 1% tyrosine from about 3% to about 8% glycine from about 3.5% to about 5.5% glutamic acid from about 2% to about 1% aspartic acid from about 0% to about 1% cysteine by weight based on the total weight of said biologically active amino acids in said mixture. 3. Antiatherosclerotic agent according to claim 1, characterized in that arginine is present in said mixture in the largest percentage by weight based on the total weight of said biologically active amino acids in said mixture. 4. Antiatherosclerotic agent according to claim 1, characterized in that said biologically active amino acids consist of their left-handed form or their crystalline form. 5. mixture useful in the treatment of atherosclerosis, characterized by an active antiatherosclerotic mixture consisting mainly of a mixture of biologically active amino acids comprising at least isoleucine, leucine, valine and arginine present in said mixture in an effective amount, whereby parenteral administration of said mixture in form of an aqueous solution for an effective period of time results in a substantial lowering of plasma cholesterol and the substantial repulsion of arterial plaque deposition and degenerative changes associated with atherosclerosis. 6. Mixture according to claim 5, characterized in that arginine is present in said mixture in the largest percentage by weight based on the total weight of said biologically active amino acids in said mixture. 7. Mixture according to claim 6, characterized in that said mixture further comprises lysine, methionine, phenylalanine, threonine, tryptophan and histidine. 8. Mixture according to claim 7, characterized in that said mixture further comprises alanine, proline, serine, tyrosine, glycine, glutamic acid, aspartic acid and cysteine. 9. Mixture according to claim 5 or 6, characterized in that said biologically active amino acids comprise their levorotatory form or their crystalline form. 10. Mixture according to claim 5 or 6, characterized in that said mixture further comprises the following mixture of amino acids: from about 3% to about 10% isoleucine from about 3% to about 10% leucine from about 2.5% to about 7.5% lysine from about 2.5% to about 7.5% methionine from about 3% to about 10% phenylalanine from about 1% to about 4% threonine from about 1% to about 3% tryptophan from about 3% to about 10% valine from about 2% to about 20% alanine from about 7% to about 20% arginine from about 1% to about 4% histidine from about 0% to about 10% proline from about 0% to about 8% serine from about 0% to about 1% tyrosine from about 0% to about 10% glycine from about 0% to about 6% glutamic acid from about 0% to about 6% aspartic acid from about 0% to about 1% cysteine by weight based on the total weight of said biologically active amino acids in said mixture. 11. Mixture according to claim 10, characterized in that said mixture further comprises: from about 4% to about 10% isoleucine from about 5% to about 10% leucine from about 4.5% to about 6.5% lysine from about 3.5% to about 5.5% methionine from about 4.5% to about 7.5% phenylalanine from about 2% to about 3.5% threonine from about 1% to about 2.5% tryptophan from about 4% to about 9% valine from approximately 7% to approximately 16% alanine from about 9% to about 18% arginine from about 15% to about 3.5% histidine from about 4.5% to about 8.5% proline from about 2.5% to about 6.5% serine from about 0% to about 1% tyrosine from about 3% to about 8% glycine from about 3.5% to about 5.5% glutamic acid from about 2% to about 4% aspartic acid from about 0% to about 1% cysteine by weight based on the total weight of said biologically active amino acids in said mixture. 12. Mixture according to any of claims 5-11, characterized in that said mixture is contained in an added dissolved substance which combines with an aqueous base and gives said aqueous solution. 13. Mixture according to claim 5, 6 or 12, which further comprises a nutritional supplement. 14. Mixture according to claim 12, characterized in that said solvent comprises a sterile, ion-free water base. 15. Mixture according to claim 12, characterized in that said added substance for dissolution is added to said aqueous base in amounts that give a concentration of at least approx. 4 g, preferably from approx. 7.5 to approx. 12 g, and most preferably from approx. 9 to approx. 11 g, of said biologically active amino acids per 100 ml solution.