Adenosine triphosphate (ATP) deficiency is the cause of many autoimmune, muscle and bone wasting disorders in humans and animals. Ratite muscle and bone extracts provide a rich source of ATP and regulatory proteins which can be used in treating conditions associated with ATP deficiency.

 

U.S. Class:	424/548
Intern'l Class:	A61K 035/34
Field of Search:	424/548

1. Field of the Invention

The present invention relates to the recognition that adenosine triphosphate (ATP) deficiency is the cause of many autoimmune, muscle and bone wasting disorders. It also relates to the discovery of ratite extracts and the use of them as a source of adenosine triphosphate (ATP) and regulatory proteins in the treatment of conditions associated with ATP deficiency in humans and animals.

2. Brief Description of the Prior Art

In allopathic medicine, diseases are treated with drugs and therapies directed specifically to the symptoms. In Darwinian or evolutionary medicine, the symptoms are addressed in a holistic manner to pinpoint a root cause and, by reverse engineering, the disease is diagnosed and treated based on the body's symptom-response mechanisms. As this applies to ATP deficiencies, there is competition for ATP in a human's or animal's body and in some instances there is not enough. For example, ATP is used in chelating toxins, essential for detoxification, but when used for this purpose, has a limiting effect on the amount of proteoglycans and glycosaminoglycans that are formed. Previously, problems associated with the production of chondroitin sulfate, heparin, hyaluronic acid, keratin sulfate, etc. were treated by treating the symptoms. The present invention provides an opportunity to treat the cause of these and a number of other conditions with an extract rich in ATP and regulatory proteins.

BRIEF SUMMARY OF THE INVENTION

In view of the above, it is an object of the present invention to provide a therapeutic agent as a source of ATP and regulatory proteins for treating conditions in humans or animals associated with ATP deficiency. Other objects and features of the invention will be in part apparent and in part pointed out hereinafter.

The present invention consists of a therapeutic agent derived from ratite muscle and bone by extracting ATP and bioactive regulatory proteins from the muscle or bone without inactivating them and to derivatives thereof wherein the ATP and bioactive regulatory proteins are partially or completely isolated from the muscle or bone. In the best mode disclosed herein, the muscle or bone is subjected to hydrolysis and degradation, the bioactive ATP and regulatory proteins are released to an aqueous phase where their activity as a therapeutic agent is amplified over their availability in the undegraded muscle or bone. They hydrolysis may be accomplished enzymatically or by cooking.

In a second aspect, the present invention further consists of a method for the treatment of a disease condition associated with ATP deficiency in a human or animal comprising applying topically, systemically or orally a therapeutic agent derived from ratite muscle or bone.

In either aspect of the invention, a functionally equivalent, synthetic mimic may replace the naturally occurring bioactive regulatory protein derived from ratite muscle protein. Broadly stated, disease conditions treatable with the ATP and bioactive regulatory protein obtained from ratite muscle or bone include those where the normal functioning of an organism is disturbed by some agent, the cause of which may be an infection with a pathogen, a genetic disorder or the effect of a chemical compound. In the best mode disclosed, degradation is effected by boiling ratite muscle and bone in water for a time sufficient to partially hydrolyze and destruct the protein, after which lipids are removed from the aqueous phase. ATP and bioactive regulatory proteins in the muscle or bone are released to the aqueous phase and may be dried by convection heat, freeze drying, etc., which seems to amplify their activity.

The invention summarized above comprises the extracts and therapies hereinafter described, the scope of the invention being indicated by the subjoined claims.

BRIEF DESCRIPTION OF THE DRAWING

In the accompanying drawings,

FIG. 1 shows a number of competitive pathways for ATP in an animal or human body and

FIG. 2 shows the role of toxins in ATP depletion, oxidative stress, acidosis and glycolysis.

DETAILED DESCRIPTION OF THE INVENTION

ATP is a nucleotide consisting of adenine, D-ribose and three phosphate groups. Two of the phosphates are linked by pyrophosphate bonds, hydrolysis of which results in a large change of free energy. ATP is an important source of energy and is used in the synthesis of other molecules through linked reactions. ATP is produced during glycolysis as glucose is metabolized to lactic or pyruvic acid and during the Krebs cycle as acetyl-CoA derived from pyruvate (formed by glycolysis) is oxidized to carbon dioxide. In animals, glucose is normally derived from glycogen.

When more glucose is need than the body can supply to maintain an adequate level of ATP, glucose is made from non-carbohydrate metabolic intermediates, formed when the animal's muscle protein undergoes proteolysis, in a process called gluconeogenesis. As the muscle undergoes proteolysis, it is believed that regulatory proteins having therapeutic value are also released.

Ratites comprise a group of flightless birds including Rhea americana (South American ostrich or rhea), Struthio camelus (African ostrich or ostrich) and Dromaius novahollandiae (emu). They have developed a unique evolutionary strategy to release enormous amounts of ATP and regulatory proteins from their muscles in response to stress. Rheas, for example, are very skittish and are capable of sustained speeds up to 35 MPH with less than 2% fat available in their muscles. Rheas have the ability to produce enormous amounts of ATP by proteolysis and gluconeogenesis, as above mentioned, simultaneously releasing regulatory proteins. Both the ATP and regulatory proteins are stored in the rhea's muscles. If a ratite is put under continued stress, requiring it to make ATP at the expense of muscle for an extended period of time, cachexia will result.

Wasting Syndrome Manifestations in Infant Rheas and Ostriches

Rhea americana and Struthio camelus chicks are notoriously frustrating to raise, often succumbing to a wasting disease from one of three bewildering syndromes which develop between one week and two months of age. Birds that survive to three months of age generally mature. Morbidity and mortality can be 100% (1). Since no single pathogen can be found consistently, this lead the applicant to believe that the syndrome is of metabolic origin, and perhaps a successful adaptation in the wild that becomes a suicide mechanism in confinement (i.e., the production of ATP and regulatory proteins at the expense of muscle).

The three syndromes are as follows:

1. Rubber Rhea Syndrome, which is currently believed to be unique to the South American ostrich, is characterized by severe hypophosphatemia, mildly depressed blood glucose, low thyroxine levels, with stunting, pliable bills, softened bones, and many chicks exhibiting widening of the proximal tibiotarsus, poor feathering, depleted adipose, and ultimately, death. Bone histopathology, decreased bone ash, gross lesions, and decreased serum phosphorus characterize moderate to severe rickets (2-4).

2. Post-protozoan stunting syndrome, following successful recovery of intestinal protozoan overgrowth after a course of treatment with metronidazole, shows shortened bones and bill, widening of the tibiometatarsus, hyperkeratinosis, stunting, poor feathering, rounded heads, and ascites with varying incidence of alopecia, achromotrichia, encephalomyopathy, spontaneous fractures, aneurysms, and paresis. This syndrome bears a remarkable similarity to aflatoxin poisoning. Chronic inflammation of the small intestine is apparent at necropsy (unpublished). This chronic inflammation is not exclusive to this disorder, it is so common as to be misconstrued as normal (5).

3. Fading chick syndrome, recognized by veterinarians and producers alike as a common problem in the ostrich and rhea, is characterized by extreme weight loss and muscle degeneration, lethargy, hypothermia, decreased appetite, stunting, frequent intestinal inflammation, ascites, and death, usually within the first month with or without secondary infections (6).

Problems with Current Therapies and Theories

Inconsistent chick survivability under a wide range of management systems, climates, feeding programs, and genetics pose a challenge to the commercial development of rheas and ostriches as livestock animals. These problems foster the widely accepted, but unsubstantiated view, that ratite chicks are immunosuppressed and the health problems are therefore multi-factorial (7-9). Typical efforts to solve these problems through known therapies utilizing strict biosecurity management or pharmaceutical regimens have resulted in frustration and increased production costs, threatening the ability to raise these animals competitively as food animals. Although ostriches have been raised domestically in South Africa for over 100 years as the domesticated hybrid commonly referred to as the African Black ostrich, the rhea and other subspecies of ostriches raised in the Untied States are basically ancient wild animals with a long history of successful evolutionary strategy in the wild state.

Compounding the situation is widespread disagreement and speculation among veterinarians regarding various causative disease organisms, which upon closer examination of the symptoms, suggests these "diseases" are variations of "Fading Chick Syndrome" with an opportunistic bacterial invader of questionable pathogenicity, Megabacteriosis (10, 11), or viruses such as coronaviral enteritis (12), rotavirus, or adenovirus (13, 14). Recent results from the experimental trials at University of Georgia show that two specific adenovirus isolates are non-pathogenic to ostriches, establishing that the presence or detection of adenovirus does not implicate pathogenicity (15), refuting prior claims of adenovirus as the leading cause of "Fading Chick Syndrome" (16). It may prove to be a synergistic virus, playing a role in increased adiposity if chickens testing positive for adenovirus. This may be a beneficial synergistic reaction to the birds in a toxin situation, increasing available adipose sites for toxin deposition (13). Therefore, the cause of Fading Chick Syndrome and Rubber Rhea Syndrome is most likely a metabolic imbalance initiated by exposure to environmental toxins often compounded by stress. This is corroborated by reduced weight gain demonstrated by exposure to the feed mycotoxin vomitoxin (16). The nature of the imbalance and the attempt by the birds to restore homeostasis provides the basis for utilizing the muscle and bone extract of the ratites in humans and animals as a therapeutic agent. All affected birds share two common disease symptoms: extreme muscle degeneration and adipose depletion. Prior reports suggest the adipose has anecdotal topical anti-inflammatory activity in humans (17-22), and compromised rhea chicks were successfully treated with adipose replacement therapy by intraperitoneal injection (23).

Present Therapy

All of the symptoms in the chicks can be attributed to hypofunctional endoplasmic reticulum, ATP depletion, and electrolyte and fatty acid imbalance caused by toxins. Correspondingly, the positive results seen in humans may be due to increased ATP and chelation for further detoxification, and improved functioning of calcium signaling and the endoplasmic reticulum. The endoplasmic reticulum regulates and synthesizes many enzymes, hormones, and lipids, and may be particularly sensitive in the young ratite chick which rely on their fat and muscle to detoxify and restore homeostasis. Their toxin sensitivity is seen in skyrocketing liver enzymes in affected chicks indicative of major liver stress and damage. The odd mechanisms of the ostrich and rhea immune systems provide the basis for a powerful therapeutic dietary supplement from the dried muscle and bone extract appropriate for humans and other animals. As shown in FIG. 2, toxins play a pivotal role in the autoimmune diseases as well as immune function and the muscle and bone extract can be used to detoxify affected individuals and animals.

Applicant is the first to recognize that the disorders of these chicks can be directly traced back to their unique immune systems that rely not only on a bioactive, anti-inflammatory body fat, but muscle components critical to their immune system function:

Muscle proteins that are interchangeable with and can act as regulatory proteins

Enzymes

Energy molecules such as ATP and phosphocreatinine

Arachidonic acid and other components of cell phospholipids

Ability to chelate toxins by an unknown system

Coupled with the recognition of the root cause of the problem was the discovery that dried ratite muscle and bone extract can be used in the treatment of conditions associated with ATP deficiency in ratites and in humans and animals generally. The ATP and regulatory proteins are available to humans and animals ingesting the extract, which supplements are not available by eating greater quantities of ratite meat as food. Other routes of administration include intravenous, intramuscular, intradermal, subcutaneous, intraperitoneal or suppository.

In addition, ratite muscle contains comparatively high levels of arachidonic acid which is a precursor in the biosynthesis of prostaglandins, thromboxanes and leukotrienes. While this might seem to be a negative, the leukotrienes in the presence of ATP chelate toxins, detoxifying the human or animal ingesting the extract.

The Mechanism of Action

The form of the proteins in the ratite extract enhances the calcium signaling system, the high level of ATP in the muscle provides energy, and the high level of arachidonic acid acts as a detoxifying agent of ratites, humans, and other animals. By a yet unknown mechanism the extract has chelating properties, perhaps through the heavy metal affinity of calmodulin.

The process of extracting and concentrating the proteins, ATP, and arachidonic acid permits use of the product as an oral supplement, although other manners of administration such as intravenously, intraperitoneal, intramuscularly, or intradermally may be successful and should not be precluded.

Given the wide range of effects seen in a large variety of symptoms and syndromes, the therapeutic mechanism effects ubiquitous factors. These factors include muscle proteins that are interchangeable with regulatory proteins, energy molecules in the form of ATP and phophocreatinine, and arachidonic acid.

Adenosine triphosphate (ATP) and the other nucleotides are affected by:

Ultraviolet light

Alternating current and electromagnetic fields

Toxins compromising their production and utilization

Nutritional and lifestyle habits

Microbial load

Free radical load

Stress

Adenosine Triphosphate (ATP) and the other nucleotides affect:

Movement of muscles by providing the energy source

ATP activates muscle components

Intercellular movement by providing the energy source

Active transport

Hormones through the cAMP second messenger system

When adenylate cyclase is activated by free radicals, prostaglandins, neurotransmitters, or hormones, ATP is catalyzed to cyclic AMP (cyclic adenosine 3, 5 monophosphate) initiating a cascade of mechanisms.

Detoxification

Used significantly in sulfation of xenobiotics (24-33)

Sulfation of proteoglycans

Required for regeneration of cartilage, bone, skin, cornea, blood vessels, heart, basement membranes in kidneys, lungs, GI tract, and mast cells which release heparin and histamine

Neurotransmission

Growth, degeneration, and regeneration of cells and organelles

Electrolyte regulation

Enzyme activation

Major biochemical pathways

The Actions of the Regulatory Muscle Proteins Include:

Plasma membrane transport and regulation including neurotransmitters

Immune system including cytokines and T cells,

Endocrine system

Endoplasmic reticulum, primarily in hepatocytes, which control:

Lipid and steroid biosynthesis, metabolism, and free radical generation

Xenobiotic detoxification of environmental toxins, drugs, and alcohol

Glycolysis and substrate cycling via glucose-6-phosphatase and calcium/calmodulin

Gluconeogenesis

Ca2+ storage and release

Protein synthesis, folding, assembly, and storage

ATP: Adenosine Triphosphate

ATP and ATP Precursors Stored in the Muscle

Ratites have the ability to manifest high energy almost instantaneously, reaching and sustaining speeds of 35 MPH. With strength to match speed, restraining a 50 pound rhea can require four able bodied young men. This energy is not lipid derived, their muscle has less than 2% fat, so this is not a major contributor to their abundant energy. In tests measuring ATP levels in muscle ratite muscle was very high and had sustained ATP levels.

The test measured relative light units (RLU at 10.sup.-9) produced by the enzymatic interaction of the ATP available in the samples and the luciferase enzyme, or firefly enzyme. ATP energy is released as light energy in the presence of this enzyme. The amount of light produced is dependent on the amount of ATP available. The measurement is done using a luminometer. 0.2 gram samples of identically prepared meat extracts were mixed with 4 mls of sterile, distilled water. 50 microliters of luciferase enzyme was added and the cuvette was sealed in the luminometer.

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    ATP levels of various extracts
                    Initial ATP decline
    Extract         Level   after 30 seconds
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    Rhea            133,000 -25%
    Beef             57,000 -32%
    Pork             78,000 -50%
    Chicken         148,700 -59%
    Chicken Baby Food	0
                  
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