Abstract:
The invention provides methods and compositions that include a nutraceutical supplement, antibiotic, and metal chelating agent that is administered to a patient to treat or prevent pathological calcification and or plaque formation as associated with Nanobacteria Calcifying Nano-Particles and/or diseases caused there-from, The method includes the administration of a therapeutically effective nutraceutical supplement, tetracycline HCL, and ethylenediaminetetraacetic acid calcium di-sodium salt to a patient in order to prevent and treat calcific disease.

Description:
CROSS REFERENCE TO RELATED APPLICATIONS  
       [0001]     This application claims priority from U.S. Provisional Patent Application Ser. No. 60/587,871, filed Jul. 15, 2004, the disclosure of which is hereby incorporated by reference in its entirety. 
     
    
     FIELD OF INVENTION  
       [0002]     This invention relates, generally, to therapeutic methods and compositions for the treatment of calcification and/or plaque-based conditions and/or diseases associated with the presence of extraneous agents in human and animal blood, sera, or other fluids comprised of self-replicating calcium phosphate macromolecular complexes termed Nanobacteria or Calcifying Nano-Particles. The methods of treatment and compositions include the combination of nutritional supplements, vitamins, herbal supplements, antibiotics, and metal chelators used separately or, more preferably, in concert.  
       BACKGROUND OF THE INVENTION  
       [0003]     Biomineralization refers, generally to the formation of discrete and organized inorganic crystalline structures within macromolecular extra cellular matrices, including, for example, the formation of calcium phosphate or crystalline hydroxy apatite. Calcification is a biomineralization process in which calcium phosphate is deposited in tissue.  
         [0004]     Examples of normal, healthy calcification include the formation of mammalian bone and dental enamel. Pathological calcification, however, has been observed to characterize a number of diseases, including but not limited to, for example, heart or circulatory diseases such as Arteriosclerosis, Atherosclerosis, Coronary Heart Disease, Chronic Heart Failure, Valve Calcifications, Arterial Aneurysms, Calcific Aortic Stenosis, Transient Cerebral Ischemia, Stroke, Peripheral Vascular Disease, Monckeberg&#39;s Disease, Vascular Thrombosis; Dental Diseases such as Dental Plaque, Gum Disease (dental pulp stones), calcification of the dentinal papilla, and Salivary Gland Stones; Chronic Infection Syndromes such as Chronic Fatigue Syndrome; Kidney and Bladder Stones, Gall Stones, Pancreas and Bowel Diseases such as Pancreatic Duct Stones, Crohn&#39;s Disease, Colitis Ulcerosa; Blood disorders; Adrenal Calcification; Liver Diseases such as Liver Cirrhosis and Liver Cysts; Testicular Microliths, Chronic Calculous Prostatitis, Prostate Calcification, Calcification in Hemodialysis Patients, Malacoplakia; Autoimmune Diseases such as Lupus Erythematosous, Schleroderma, Dermatomyositis, Cutaneous polyarteritis, Panniculitis (Septal and Lobular), Antiphospholipid Syndrome, Arteritis Nodosa, Thrombocytopenia, Hemolytic Anemia, Myelitis, Livedo Reticularis, Chorea, Migraine, Junvenile Dermatomyositis, Graves Disease, Chronic Thyroiditis, Hypothyreoidism, Type 1 Diabetes Mellitis, Addison&#39;s Disease, and Hypopituitarism; Placental and Fetal Disorders, Polycystic Kidney Disease, Glomerulopathies; Eye Diseases such as Corneal Calcifications, Cataracts, Macular Degeneration and Retinal Vasculature-derived Processes and other Retinal Degenerations; Retinal Nerve Degeneration, Retinitis, and Iritis; Ear Diseases such as Otosclerosis, Degeneration of Otoliths and Symptoms from the Vestibular Organ and Inner Ear (Vertigo and Tinnitus); Thyroglossal cysts, Thyroid Cysts, Ovarian Cysts; Cancer such as Meningiomas, Breast Cancer, Prostate Cancer, Thyroid Cancer, Serous Ovarian Adenocarcinoma; Skin diseases such as Calcinosis Cutis, Skin Stones, Calciphylaxis, Psoriasis, Eczema, Lichen Ruber Planus or Lichen Simple Cysts; Choroid Plexus Calcification, Neuronal Calcification, Calcification of the Falx Cerebri, Calcification of the Intervertebral Cartilage or Disc, Intercranial or Cerebral Calcification, Rheumatoid Arthritis, Calcific Tenditis, Oseoarthritis, Fibromyalgia, Bone Spurs, Diffuse Interstitial Skeletal Hyperostosis, Intracranial Calcifications such as Degenerative Disease Processes and Dementia; Erythrocyte-Related Diseases involving Anemia, Intraerythrocytic Nanobacterial Infection and Splenci Calcifications; Chronic Obstructive Pulmonary Disease, Broncholiths, Bronchial Stones, Neuropathy, Calcifications and Encrustations of Implants, Mixed Calcified Biofilms, and Myelodegenerative Disorders such as Multiple Sclerosis, Lou Gehrig&#39;s, and Alzheimer&#39;s Disease. Although the cause of pathological calcification remains unknown, it has been observed that each of the foregoing conditions is often associated with the presence of a very small, mineral-associated bacteria-like self-replicating calcium-phosphate macromolecular complexes forms termed Nanobacteria (Nanobacterium sanguineum) or Calcifying Nano-Particles which are known for their ability to create calcium phosphate coated vesicles or nanoparticles that multiply in blood and in cell culture medium like living cells. Nanobacteria (“NB”) or Calcifying Nano-Particles (“CNP”) are approximately 20-200 nanometers in size and are currently the smallest known self-replicating particles or bacteria.  
         [0005]     NB/CNP-induced calcification results from the formation of calcium-phosphate mineral deposits around each calcifying nano-particle. NB/CNPs secrete a protective calcific biofilm (i.e., a lipopolysaccharide (LPS) endotoxin biofilm) that also allows multiple NB/CNP to connect, collaborate and apparently form “colonies.” This calcific biofilm also allows the NB/CNP to expand, contract and move. The biofilm appears to be generated as part of a stress response mechanism; it is primarily observed, for example, when NB/CNP are chemically, physiologically or environmentally attacked, when they are working together and/or during NB/CNP reproduction. The biofilm that is secreted by the NB/CNP is a potent endotoxin and activates a thrombic cascade causing inflammation, swelling and the release of cytokines, interleukins, leukocytes, mast cells, collagenase, matrix metalloproteinases and other immune-response events in surrounding cells.  
         [0006]     NB/CNP are “extremeophiles” (i.e., highly tolerant to heat, freezing, dehydration and Gamma Irradiation) and are apparently more resistant than most bacteria to destruction. Thus, NB/CNPs have been found to be residual contaminants on otherwise sterilized medical products such as tissue, blood and bovine serum. Similarly, NB/CNP cannot be killed using most antibiotics, including, for example, Penicillin, Cephalosporins, or Macrolides. It has been observed, however, that NB/CNP are sensitive to in vitro treatment with certain tetracycline&#39;s and that EDTA can assist in dissolving the protective biofilm secreted by NB/CNP.  
         [0007]     Bench and clinical research have established that atherosclerosis is an inflammatory disease characterized by injury or infection of the vascular endothelium resulting in the formation of atheromas and pathological calcification. Inflammatory cascade responses within individual atheromas (as the immune system attempts to “wall off” or isolate an area of injury) result in the synthesis of a fibro-lipid matrix synthesis and the degradation/absorption of soft plaques. The rate of plaque synthesis-resorption is dependent upon the degree and/or stage of inflammatory activity within atheroma. Mature atheromas, for example, contain pathological calcification deposits that have been observed to increase at an annual rate of 24-82%.  
         [0008]     Although pathological calcification deposits are a hallmark of atherosclerosis, the precise mechanism of such calcium precipitation has remained elusive. It has been widely speculated, however, that NB/CNP play a critical role in the pathological calcification processes associated with atherosclerosis. In particular, NB/CNP have been detected in atherosclerotic plaques, calcified carotid arteries, aortic aneurysms and cardiac valves. Furthermore, NB/CNP particles morphologically and functionally resemble the calcifiable vesicles, are capable of active calcium phosphate precipitation under suitable nutrient conditions and have previously been isolated from atherosclerotic aorta  
         [0009]     Accordingly, there is a need for a specialized treatment comprising appropriate combinations of compositions of at least one of tetracycline, EDTA, and other materials for the treatment of NB/CNP-based atherosclerotic disease. A further objective of the invention is to identify a treatment protocol that is effective for treatment of atherosclerotic disease and which includes in vitro treatment with tetracycline, EDTA and other materials.  
       SUMMARY OF THE INVENTION  
       [0010]     The invention provides therapeutic methods and compositions for the treatment of calcification and/or plaque-based conditions associated with nanobacteria/calcifying nano-particle infection and in particular, atherosclerotic disease. Such therapeutic methods and compositions include administering a combination of a nutraceutical powder, tetracycline HCl and ethylenediaminetetraacetic acid disodium salt (EDTA-sequestrant).  
         [0011]     Thus, one aspect of the invention is to provide compositions for the treatment of a disease characterized by calcification and/or plaque formation, or for the treatment of pathological calcification caused by Nanobacteria Calcifying Nano-Particles comprising at least one of a nutraceutical supplement, an antibiotic, and a metal chelator.  
         [0012]     In accordance with the invention, the nutraceutical supplement may be comprised of a mixture of one ore more of Niacin, Vitamin B6, Folate, Vitamin C, Selenium, L-Arginine, L-Ornithine, L-Lysine, Bromelain, Trypsin, Papain, Co-Q10, Grapeseed Extract, Hawthorne Berry, Vitamin A, Vitamine E, Vitamin B1, Vitamin B2, Vitamin B12, Magnesium Citrate, Methyl Sulfonyl Methane,  Curcuma Longa , Quercitin, Pycnogenol, Gugulipid.  
         [0013]     In accordance with the invention, the antibiotic may be comprised of at least one of tetracycline, tetracycline HCL, Chlortetracycline, Democlocycline, Doxycycline, Methacycline, Oxytetracycline, Rolitetracycline, Minocycline, Sancycline, or salts thereof.  
         [0014]     In accordance with the invention, the metal chelator may be comprised of at least one or more of Ethylenediaminetetraacetic acid (EDTA), Ethyleneglycoltetraacetic acid (EGTA), Diethylenetriaminepentaacetate (DTPA), Hydroxyethylethylenediaminetriacetic acid (HEEDTA), Diaminocyclohexanetetraacetic acid (CDTA), 1,2-Bis(2-aminophenoxy)ethane-N,N,N′,N′-tetraacetic acid (BAPTA), and pharmaceutically acceptable salts thereof.  
         [0015]     In yet another aspect, the present invention relates to a method of using a composition comprising calcium chelators, bisphosphonates and/or citrate compounds which comprises administering said composition to reduce and/or prevent calcification related diseases, such as heart or circulatory diseases such as Arteriosclerosis, Atherosclerosis, Coronary Heart Disease, Chronic Heart Failure, Valve Calcifications, Arterial Aneurysms, Calcific Aortic Stenosis, Transient Cerebral Ischemia, Stroke, Peripheral Vascular Disease, Monckeberg&#39;s Disease, Vascular Thrombosis; Dental Diseases such as Dental Plaque, Gum Disease (dental pulp stones), calcification of the dentinal papilla, and Salivary Gland Stones; Chronic Infection Syndromes such as Chronic Fatigue Syndrome; Kidney and Bladder Stones, Gall Stones, Pancreas and Bowel Diseases such as Pancreatic Duct Stones, Crohn&#39;s Disease, Colitis Ulcerosa; Blood disorders; Adrenal Calcification; Liver Diseases such as Liver Cirrhosis and Liver Cysts; Testicular Microliths, Chronic Calculous Prostatitis, Prostate Calcification, Calcification in Hemodialysis Patients, Malacoplakia; Autoimmune Diseases such as Lupus Erythematosous, Schleroderma, Dermatomyositis, Cutaneous polyarteritis, Panniculitis (Septal and Lobular), Antiphospholipid Syndrome, Arteritis Nodosa, Thrombocytopenia, Hemolytic Anemia, Myelitis, Livedo Reticularis, Chorea, Migraine, Junvenile Dermatomyositis, Graves Disease, Chronic Thyroiditis, Hypothyreoidism, Type 1 Diabetes Mellitis, Addison&#39;s Disease, and Hypopituitarism; Placental and Fetal Disorders, Polycystic Kidney Disease, Glomerulopathies; Eye Diseases such as Corneal Calcifications, Cataracts, Keratopathy, Macular Degeneration and Retinal Vasculature-derived Processes and other Retinal Degenerations; Retinal Nerve Degeneration, Retinitis, and Iritis; Ear Diseases such as Otosclerosis, Degeneration of Otoliths and Symptoms from the Vestibular Organ and Inner Ear (Vertigo and Tinnitus); Thyroglossal cysts, Thyroid Cysts, Ovarian Cysts; Cancer such as Meningiomas, Breast Cancer, Prostate Cancer, Thyroid Cancer, Serous Ovarian Adenocarcinoma; Skin diseases such as Pyoderma gangrenosum, Dermatomyositis, eccrine sweat duct calcification, trichoepithelioma, pilomatrixoma, necrobiosis lipoidica, Calcinosis Cutis, Skin Stones, Calciphylaxis, Psoriasis, Eczema, Lichen Ruber Planus or Lichen Simple Cysts; Choroid Plexus Calcification, Neuronal Calcification, Calcification of the Falx Cerebri, Calcification of the Intervertebral Cartilage or Disc, Intercranial or Cerebral Calcification, Rheumatoid Arthritis, Calcific Tenditis, Oseoarthritis, Fibromyalgia, Bone Spurs, Diffuse Interstitial Skeletal Hyperostosis, Intracranial Calcifications such as Degenerative Disease Processes and Dementia; Erythrocyte-Related Diseases involving Anemia, Intraerythrocytic Nanobacterial Infection and Splenci Calcifications; Chronic Obstructive Pulmonary Disease, Broncholiths, Bronchial Stones, Neuropathy, Calcifications and Encrustations of Implants, Mixed Calcified Biofilms, and Myelodegenerative Disorders such as Multiple Sclerosis, Lou Gehrig&#39;s, and Alzheimer&#39;s Disease in an individual in need thereof.  
         [0016]     Yet another aspect of this invention is to provide methods for administering a pharmaceutically or therapeutically effective amount of a composition of the invention to a human or mammal. 
     
    
     DETAILED DESCRIPTION OF THE INVENTION  
       [0017]     This application incorporates herein by reference in its entirety the copending and commonly assigned U.S. Non-provisional patent application Ser. No. 10/891,483 entitled “Methods and Compositions for the treatment of Diseases Characterized by Pathological Calcification” (Attorney Docket No. 19772-0004) which was filed with the United States Patent and Trademark Office on Jul. 15, 2004.  
         [0018]     As discussed above, nanobacteria/calcifying nano-particle (“NB/CNP”) cause pathological calcification associated with a number of conditions, including atherosclerotic disease. Thus, an objective of the invention is to provide compositions useful in countering such NB/CNP-associated pathological calcification. Similarly, another objective of the invention is to provide a protocol for administering such compositions for the treatment of atherosclerotic diseases.  
         [0019]     The invention provides therapeutic methods and compositions for the treatment of calcification and/or plaque-based conditions associated with NB/CNP infection and atherosclerotic disease. In particular, the invention includes compositions and therapeutic protocols for administering such compositions that include a nutraceutical powder, certain tetracyclines and ethylenediaminetetraacetic acid calcium disodium salt (EDTA-sequestrant). The combination of these ingredients also offers novel compositions that may be useful in the treatment of other NB/CNP related/pathological calcification conditions, including but not limited to, for example, heart or circulatory diseases such as Arteriosclerosis, Atherosclerosis, Coronary Heart Disease, Chronic Heart Failure, Valve Calcifications, Arterial Aneurysms, Calcific Aortic Stenosis, Transient Cerebral Ischemia, Stroke, Peripheral Vascular Disease, Monckeberg&#39;s Disease, Vascular Thrombosis; Dental Diseases such as Dental Plaque, Gum Disease (dental pulp stones), calcification of the dentinal papilla, and Salivary Gland Stones; Chronic Infection Syndromes such as Chronic Fatigue Syndrome; Kidney and Bladder Stones, Gall Stones, Pancreas and Bowel Diseases such as Pancreatic Duct Stones, Crohn&#39;s Disease, Colitis Ulcerosa; Blood disorders; Adrenal Calcification; Liver Diseases such as Liver Cirrhosis and Liver Cysts; Testicular Microliths, Chronic Calculous Prostatitis, Prostate Calcification, Calcification in Hemodialysis Patients, Malacoplakia; Autoimmune Diseases such as Lupus Erythematosous, Schleroderma, Dermatomyositis, Cutaneous polyarteritis, Panniculitis (Septal and Lobular), Antiphospholipid Syndrome, Arteritis Nodosa, Thrombocytopenia, Hemolytic Anemia, Myelitis, Livedo Reticularis, Chorea, Migraine, Junvenile Dermatomyositis, Graves Disease, Chronic Thyroiditis, Hypothyreoidism, Type 1 Diabetes Mellitis, Addison&#39;s Disease, and Hypopituitarism; Placental and Fetal Disorders, Polycystic Kidney Disease, Glomerulopathies; Eye Diseases such as Corneal Calcifications, Cataracts, Macular Degeneration and Retinal Vasculature-derived Processes and other Retinal Degenerations; Retinal Nerve Degeneration, Retinitis, and Iritis; Ear Diseases such as Otosclerosis, Degeneration of Otoliths and Symptoms from the Vestibular Organ and Inner Ear (Vertigo and Tinnitus); Thyroglossal cysts, Thyroid Cysts, Ovarian Cysts; Cancer such as Meningiomas, Breast Cancer, Prostate Cancer, Thyroid Cancer, Serous Ovarian Adenocarcinoma; Skin diseases such as Calcinosis Cutis, Skin Stones, Calciphylaxis, Psoriasis, Eczema, Lichen Ruber Planus or Lichen Simple Cysts; Choroid Plexus Calcification, Neuronal Calcification, Calcification of the Falx Cerebri, Calcification of the Intervertebral Cartilage or Disc, Intercranial or Cerebral Calcification, Rheumatoid Arthritis, Calcific Tenditis, Oseoarthritis, Fibromyalgia, Bone Spurs, Diffuse Interstitial Skeletal Hyperostosis, Intracranial Calcifications such as Degenerative Disease Processes and Dementia; Erythrocyte-Related Diseases involving Anemia, Intraerythrocytic Nanobacterial Infection and Splenci Calcifications; Chronic Obstructive Pulmonary Disease, Broncholiths, Bronchial Stones, Neuropathy, Calcifications and Encrustations of Implants, Mixed Calcified Biofilms, and Myelodegenerative Disorders such as Multiple Sclerosis, Lou Gehrig&#39;s, and Alzheimer&#39;s Disease.  
         [0020]     The nutraceutical powder includes Vitamin C, Vitamin B6, Niacin, Folic Acid, Selenium, EDTA, L-Arginine, L-Lysine, L-Ornithine, Bromelain, Trypsin, Niacin, CoQ10, Grapeseed Extract, Hawthorn Berry and Papain. The nutraceutical powder can also include other ingredients and materials as described below. The quantity of each component of the nutraceutical powder as well as the quantity of nutraceutical powder used in the invention may be varied for different patients and/or treatment conditions. For instance, the addition of other vitamins such as, but not limited to, Vitamin A as β Carotene, Vitamin E as d-alpha Tocopherol Succinate, Vitamin B 1 as thiamine mononitrate, Vitamin B2 as riboflavin, and Vitamin B 12 as Cyanocobalamin. Other ingredients such as Methyl Sulfonyl Methane, Magnesium Citrate, Zinc Citrate, and herbal extracts such as  Mahonia aquifolium, Curcuma Longa , Quercetin, picnogenol, gugulipid, Schizandra chinensis, Licorice root, Alfalfa seed, wheatgrass, green barley grass,  Chorella algae, Spirulina , Flaxseed, milk thistle, and/or Aslanguanda may be added, Other enzymes and or amino acids may also be added to the formulation such as, but not limited to, Lipase, Protease, Peptase, Serrapeptase, Cellulase, L-Glutathione.  
         [0021]     Suitable tetracycline&#39;s include, but are not limited to, tetracycline, tetracycline HCl, chlortetracycline, demeclocycline, doxycycline, methacycline, oxytetracycline, rolitetracycline, minocycline, sancycline and pharmaceutically acceptable salts thereof. A preferred tetracycline is tetracycline HCl. The dose of these medicines may be varied for different patients and/or treatment conditions.  
         [0022]     Suitable chelating agents include, but are not limited to one or more of Ethylenediaminetetraacetic acid (EDTA), Ethyleneglycoltetraacetic acid (EGTA), Diethylenetriaminepentaacetate (DTPA), Hydroxyethylethylenediaminetriacetic acid (HEEDTA), Diaminocyclohexanetetraacetic acid (CDTA), 1,2-Bis(2-aminophenoxy)ethane-N,N,N′,N′-tetraacetic acid (BAPTA), and pharmaceutically acceptable salts thereof.  
         [0023]     One hundred patients with stable coronary artery disease (“CAD”) and positive coronary artery calcium (“CAC”) scores were initially enrolled in a four month treatment regimen that included daily administration of a three-component composition composed of the nutraceutical powder (discussed above), tetracycline HCl and ethylenediaminetetraacetic acid calcium disodium salt (EDTA-sequestrant). Exclusion criteria included: (1) known tetracycline allergy, (2) zero CAC score, (3) recent (&lt;30 days) major adverse cardiac event, (4) women of childbearing age, (5) recent diagnosis of thyroid or parathyroid disease, (6) clinically significant renal insufficiency or liver function abnormalities and (7) recent (&lt;30 days) acute congestive heart failure. CAC scoring was repeated at four months and serum samples were analyzed for NB/CNP antigen and baseline serology at zero, two and four months. Complete blood count, metabolic panel, liver function, C-reactive protein (hs-CRP) and lipids were analyzed at zero and four months. Other than discontinuing any herbal or vitamin preparation, patients maintained their normal medical regime during the study. Baseline History and Physical examination were performed. The same CAC scoring machine was used for each individual patient to assess initial and final CAC scores. CAC scoring radiologists were experienced in CAC scoring and were blinded to patient identity. CAC scoring was repeated after four months of treatments. Before completion of the study, one patient withdrew secondary to a presumed sensitivity to tetracycline HCL and twenty-two patients were withdrawn due to noncompliance.  
         [0024]     As discussed in more detail below in conjunction with the accompanying Tables, 100% of the seventy-seven patients completing the study were positive for NB/CNP serology, antigen or both. Responders (n=44; 57%) had significant decreases in total CAC scores (p=0.001); the average decrease being 14%. Non-responders (n=33; 44%) had no change or had increases in CAC scores. No adverse physiologic effects were seen in the renal, hepatic, or hematopoetic systems of the treated patients. Angina was decreased or ablated in 16 of 19 patients (84%). Lipid profiles significantly improved in the non-atherogenic direction (p=0.001). Such a change in the lipid profiles is significant given that 86% of the patient group were on continuous statin medication prior to treatment.  
         [0025]     In the accompanying Tables, data is presented as frequency and percentage distributions. Values for continuous variables are expressed as mean plus or minus a (“±”) standard deviation. Within group comparisons of initial and ending CAC scores (mean) and laboratory values were conducted using a paired t-test. Between group comparisons of continuous variables were conducted with the Student&#39;s t-test. Univariate analysis of selected discrete variables was accomplished by X 2 , the continuity X 2  analysis or a two-tailed Fischer Exact test with the appropriate degrees of freedom. Statistical procedures were performed using the Number Cruncher Statistical Systems® (NCSS, Kaysville, Utah). Ap-value of less than or equal to 0.05 was designated as statistically significant in the treatment study.  
         [0026]     Tables 1 and 2 provide statistical data and physical characteristics of participants in a study evaluating the clinical effects of the invention. In Table 1, the initial physical and clinical characteristics of the final study participants (n=77) are described. In Table 2, the seventy-seven participants are subdivided into “responder” and “nonresponder” groups (as defined below) based on their response to treatment with the invention. Table 2 further illustrates the pretreatment physical characteristics for both the “responder” and “nonresponder” groups. The data in Table 2 indicates that both groups had comparable pre-treatment clinical variables and risk factors.  
         [0027]     Table 3 demonstrates that 44 (57%) of the seventy-seven patients responded to treatment with the invention as evidenced by a decrease in total CAC score. The remaining 33 patients (43%) were considered “nonresponders” based solely on their CAC score. As can be seen in Table 3, total CAC scores decreased significantly (p=0.001) from the beginning to the end of the study for the responder group. Significant reduction in both the left anterior descending coronary artery and the right coronary artery CAC scores were also documented p=0.002). There was no significant difference found in the left main coronary artery (p=0.972) or circumflex coronary artery CAC scores (p=0.106).  
         [0028]     Table 4 illustrates that all responder group patients tested positive for the presence of anti-NB/CNP IgG antibodies prior to the commencement of therapy. During treatment with the invention, NB/CNP antigen and serology titers tended to fluctuate (although the fluctuations were not statistically significant) in all patients independent of changes in CAC scores or stage of therapy.  
         [0029]     Table 5 demonstrates the beneficial changes in the lipid profiles for responder group patients following treatment with the invention. Notably, it was observed that responder group patients experienced reduced total cholesterol levels (p=0.001), reduced triglycerides p=0.006), decreased LDL (p=0.001) and increased HDL (p=0.001) following treatment with the invention.  
         [0030]     In addition to the favorable results illustrated in Tables 1-5, other data supports the efficacy of the invention. For example, prior to treatment, 19 patients (25%) had stable angina pectoris. Following four months of treatment, the angina symptoms had been either eliminated or substantially ameliorated in 16 of the 19 (84%) patients (p=0.013). Similarly, two patients (3%) with severe claudication and faint pedal pulses reported a diminution of claudication symptoms and the return of their peripheral pulses to normal values following treatment.  
         [0031]     The foregoing data demonstrates that administration of a combination of a nutraceutical powder, certain antibiotics and EDTA for sustained periods is effective for treating CAD patients. Specifically, every second CAD patient treated as described herein demonstrated an objective improvement in their cardiac vasculature performance and had appreciably decreased CAC scores (avg. ˜14% decrease). These results are particularly encouraging considering that CAC scores are known to increase by more than 20% annually. These results highlight the significance of the invention given that there have been no previous reports showing a significant decrease in CAC scores pursuant to any known means of intervention.  
         [0032]     Furthermore, based on the foregoing data (for both the responder and nonresponder patient groups), it is possible to infer that other variables, including, for example, treatment time, plaque density/volume, tissue penetration and blood supply may be critical factors that influence overall outcomes related to treatment efficacy. Based on these findings, it appears that CAC scores would continue to decrease in conjunction over longer periods of therapy (i.e., plaque regression over time). Additionally, the treatment regimen did not produce any apparent or significant adverse physiological effects on the study participants.  
       EXAMPLES  
       [0033]     The invention is further illustrated by the following examples. All scientific and technical terms have the meanings as understood by one with ordinary skill in the art. The specific examples that follow illustrate the methods in which the compositions of the present invention may be prepared and/or protocols for the administration of such compositions to a patient in need thereof. Such examples, however, are merely illustrative and are not intended nor should be construed as limiting the invention in sphere or scope. The methods may be adapted and/or varied in order to produce compositions embraced by this invention but not specifically disclosed. Further, variations of the methods to produce the same compositions in somewhat different fashion will be evident to one skilled in the art.  
         [0000]     1. Formulations  
         [0034]     In one embodiment of the invention, a composition for treatment of atherosclerotic diseases associated with NB/CNP infection that includes at least three components is disclosed. These components include a quantity of a nutraceutical powder (that includes Vitamin C, Vitamin B6, Niacin, Folic Acid, Selenium, EDTA, L-Arginine, L-Lysine, L-Ornithine, Bromelain, Trypsin, Niacin, CoQ10, Grapeseed Extract, Hawthorn Berry and Papain), a quantity of a tetracycline compound and a quantity of ethylenediaminetetraacetic acid disodium salt or calcium di-sodium salt (EDTA-sequestrant).  
         [0035]     In another embodiement, the nutraceutical powder may also include other Vitamins such as, but not limited to, Vitamin A, Vitamin E, Vitamin B1, B2, and B12. Materials such as methyl sulfonyl methane (MSM), Citrates such as Magnesium Citrate or Zinc Citrate and herbal extracts such as  Mahonia aquifolium, Curcuma longa  (turmeric), Lipase, Protease, Peptase, Serrapeptase, Cellulase, L-Glutathione,  Schizandra Chinensis , Licorice Root, Quercetin, Alfalfa Seed, Wheatgrass, Green Barley Grass,  Chlorella  Algae,  Spirulina , Flaxseed, Milk Thistle, picnogenol, Gugulipid, Aslaguanda may also be added to the formulae as predicated by specific patient requirements.  
         [0036]     In another embodiment, the quantity of the nutraceutical powder component is mixed with water, juice (e.g., apple or orange juice) or other suitable liquid prior to being administered.  
         [0037]     In another embodiment, the quantity of the nutraceutical powder component is 5 cm 3  and is mixed with water, juice (e.g., apple or orange juice) or other suitable liquid prior to being administered.  
         [0038]     In other embodiments, the quantity of nutraceutical powder is formulated as either a pill or capsule.  
         [0039]     In another embodiment the tetracycline compound is tetracycline HCl.  
         [0040]     In another embodiment, 500 mg of the tetracycline HCl component is formulated as a capsule before being administered.  
         [0041]     In another embodiment, 500 mg of the tetracycline HCl component is formulated as a pill before being administered.  
         [0042]     In another embodiment, 1500 mg of the ethylenediaminetetraacetic acid calcium disodium salt (EDTA-sequestrant) component is formulated as a suppository before being administered.  
         [0043]     As will be appreciated by those knowledgeable in the art, the therapeutic components of the invention may be individually or collectively formulated in different manners, quantities and/or combinations and may otherwise be used in combination with other treatments. Furthermore, the therapeutic composition of the present invention may be packaged in any convenient, appropriate packaging.  
         [0044]     In addition to the specific component formulations recited in the above examples, each component of the invention may be in various other forms suitable for oral use (for example as tablets, lozenges, hard or soft capsules, aqueous or oily suspensions, emulsions, dispersible powders or granules, syrups or elixirs), for topical use (for example as creams, ointments, gels, or aqueous or oily solutions or suspensions) for administration by inhalation (for example as a finely divided powder or a liquid aerosol), for administration by insufflation (for example as a finely divided powder) or for parenteral administration (for example as a sterile aqueous or oily solution for intravenous, subcutaneous, or intramuscular dosing), or as a suppository for rectal dosing.  
         [0045]     Suitable pharmaceutically-acceptable excipients for a tablet formulation include, for example, inert diluents such as lactose, sodium carbonate, granulating and disintegrating agents such as corn starch or alginic acid; binding agents such as starch; lubricating agents such as stearate, stearic acid, fumed silica or talc; preservative agents such as ethyl or propyl p-hydroxybenzoate, and anti-oxidants, such as ascorbic acid or tocopherol acetate. Tablet formulations may be uncoated or coated either to modify their disintegration and the subsequent absorption of the active ingredient within the gastrointestinal tract, or to improve their stability and/or appearance, in either case, using conventional coating agents or fillers such as hydroxy propyl methyl cellulose (Methocel) or other cellulose and procedures well known in the art  
         [0046]     Compositions for oral use of one or more of the components may be in the form of hard gelatin capsules in which the active ingredient is mixed with an inert solid diluent, for example, calcium carbonate, calcium sulfate dihydrate, calcium phosphate or kaolin, or as soft gelatin capsules in which the active ingredient is mixed with water or an oil such as peanut oil, liquid paraffin, or olive oil.  
         [0047]     Aqueous suspensions of one or more of the components generally contain the active ingredient in finely powdered form together with one or more suspending agents, such as sodium carboxymethylcellulose, methylcellulose, hydroxyethyl starch, starch acetate, hydroxypropylmethylcellulose, sodium alginate, polyvinyl-pyrrolidone, gum tragacanth and gum acacia; dispersing or wetting agents such as lecithin or condensation products of an alkylene oxide with fatty acids (for example polyoxethylene stearate), or condensation products of ethylene oxide with long chain aliphatic alcohols, for example heptadecaethyleneoxycetanol, or condensation products of ethylene oxide with partial esters derived from fatty acids and a hexitol such as polyoxyethylene sorbitol monooleate, or condensation products of ethylene oxide with partial esters derived from fatty acids and hexitol anhydrides, for example polyethylene sorbitan monooleate. The aqueous suspensions may also contain one or more preservatives (such as ethyl or propyl p-hydroxybenzoate, anti-oxidants (such as ascorbic acid or tocopherol acetate), coloring agents, flavoring agents, and/or sweetening agents (such as sucrose, stevia, sucralose, xylitol, saccharine or aspartame).  
         [0048]     Oily suspensions of one or more of the components may be formulated by suspending the active ingredient in a vegetable oil (such as arachis oil, olive oil, sesame oil or coconut oil) or in a mineral oil (such as liquid paraffin). The oily suspensions may also contain a thickening agent such as beeswax, hard paraffin or cetyl alcohol. Sweetening agents such as those set out above, and flavoring agents may be added to provide a palatable oral preparation. These compositions may be preserved by the addition of an anti-oxidant such as ascorbic acid or tocopherol acetate.  
         [0049]     Dispersible powders and granules suitable for preparation of an aqueous suspension of one or more of the components by the addition of water (or other suitable liquid such as juice) generally contain the recited ingredient(s) together with a dispersing or wetting agent, suspending agent and one or more preservatives. Suitable dispersing or wetting agents and suspending agents are exemplified by those already mentioned above. Additional excipients such as sweetening, flavoring and coloring agents may also be present.  
         [0050]     One or more of the components of the invention may also be in the form of oil-in-water emulsions. The oily phase may be a vegetable oil, such as olive oil or arachis oil, or a mineral oil, such as for example liquid paraffin or a mixture of any of these. Suitable emulsifying agents may be, for example, naturally-occurring gums such as gum acacia or gum tragacanth, naturally-occurring phosphatides such as soya bean, lecithin, an esters or partial esters derived from fatty acids and hexitol anhydrides (for example sorbitan monooleate) and condensation products of the said partial esters with ethylene oxide such as polyoxyethylene sorbitan monooleate. The emulsions may also contain sweetening, flavoring and preservative agents.  
         [0051]     Syrups and elixirs of one or more of the components may be formulated with sweetening agents such as glycerol, propylene glycol, sorbitol, aspartame or sucrose, and may also contain a demulcent, preservative, flavoring and/or coloring agent.  
         [0052]     One or more of the components may also be in the form of a sterile injectable aqueous or oily suspension, which may be formulated according to known procedures using one or more of the appropriate dispersing or wetting agents and suspending agents, which have been mentioned above. A sterile injectable preparation may also be a sterile injectable solution or suspension in a non-toxic parenterally-acceptable diluent or solvent, for example a solution in 1,3-butanediol.  
         [0053]     Suppository formulations of one or more of the components may be prepared by mixing the active ingredient with a suitable non-irritating excipient which is solid at ordinary temperatures but liquid at the rectal temperature and will therefore melt in the rectum to release the drug. Suitable excipients include, for example, cocoa butter, polyethylene glycols and stearates.  
         [0054]     Compositions of one or more of the components for administration by insufflation may be in the form of a finely divided powder containing particles of average diameter of, for example, 30 μm or much less, the powder itself comprising either active ingredient alone or diluted with one or more physiologically acceptable carriers such as lactose. The powder for insufflation is then conveniently retained in a capsule containing, for example, 1 to 50 mg of active ingredient for use with a turbo-inhaler device, such as is used for insufflation of the known agent sodium cromoglycate.  
         [0055]     Compositions of one or more of the components for administration by inhalation may be in the form of a conventional pressurized aerosol arranged to dispense the active ingredient either as an aerosol containing finely divided solid or liquid droplets. Conventional aerosol propellants such as volatile fluorinated hydrocarbons or hydrocarbons may be used and the aerosol device is conveniently arranged to dispense a metered quantity of active ingredient.  
         [0056]     The amount of one or more of the ingredients comprising each component of the invention can be altered or combined with one or more excipients to produce a single dosage form and each such combination may vary depending upon the host treated and the particular route of administration. For example, a formulation intended for oral administration to humans may contain a component compounded with an appropriate and convenient amount of excipients that may vary from about 0.1 to about 99% by weight of the total composition. Such dosages may be obtained by mixing each component of the invention with different excipients (as recited above) such as agglutinants, disintegrators, lubricants, sliders or fillers. Other excipients include lactose, corn starch, saccharose, stearate, microcrystalline cellulose, sodium croscarmellose gelatin, cellulose acetophtalate, titanium dioxide, fumed and precipitated silicates, special talc for tablets and polyethylene glycol.  
         [0000]     2. Treatment Protocols  
         [0057]     The invention further contemplates a protocol for administering the three-components of the invention for treatment of atherosclerotic diseases associated with NB/CNP infection. According to this aspect of the invention, there is provided a protocol for the separate and sequential administration of the nutraccutical powder (discussed above), tetracycline HCl and ethylenediaminetetraacetic acid disodium salt (EDTA-sequestrant) in sufficient quantity to an individual in need thereof. The protocol of the present invention can be administered to a patient by any available and effective delivery system including, but not limited to, oral, parenteral, transdermal, intranasal, sublingual, transmucosal, intra-arterial, or intradermal modes of administration in dosage unit formulations containing conventional nontoxic pharmaceutically acceptable carriers, adjuvants, and vehicles as desired, such as a depot or a controlled release formulation.  
         [0058]     In one embodiment of the treatment protocol, a patient is instructed, prior to going to bed, to mix approximately 5 cm 3  of the nutraceutical powder in water, juice (e.g., apple or orange juice) or other suitable liquid prior to being administered. Thereafter, the patient is instructed to orally consume the nutraceutical powder solution. In this embodiment, the patient is also instructed to orally consume approximately 500 mg of tetracycline HCl that had been formulated as a capsule before administration. Next, the patient is instructed to rectally insert approximately 1500 mg of ethylenediaminetetraacetic acid disodium salt (EDTA-sequestrant) that had been formulated as a suppository before administration. Once the three components of the composition were administered, the patient was instructed to lie down flat and fall asleep.  
         [0059]     Variations in the above treatment protocol can readily be made. In other embodiments, for example, the order in which the components are administered can be altered. Similarly, in differing embodiments, different quantities of each component may be employed and/or the components may individually or collectively formulated in different manners as warranted by prevailing conditions or patient needs.  
                                               TABLE 1                           Study Variables            Variables   Number   Percentage                    Total Study       77   100       Number       Gender   Male   62   80.5           Female   15   19.5       Age Groups   Under 50   4   5.2       (years)   50-59   25   32.5           60-69   25   32.5           70-79   20   26           80 and Over   3   3.9           Mean   63.2 ± 8.9           Range   42-81       Coronary Risk   Hypertension   52   67.5       Factors   Hyperlipidemia   68   88.3           Diabetes Mellitus   20   26.0           Peripheral Vascular Disease   10   13.0           History of Congestive Heart   5   6.5           Renal Insufficiency   2   2.6           (Creatine &gt; 2.0 mg/dL)           Previous Myocardial Infarction   14   18.1           Stable Angina   19   24.7       Previous   Prior Coronary Artery Bypass   39   50.7       Cardiovascular   Grafting           Prior Percutaneous Coronary   17   22.1           Intervention       Current   Statins   66   85.7       Medications   Nitrates   23   29.9           Anticagulants   3   3.9           Beta Blockers   42   54.6           ACE Inhibitors   28   36.4           Diuretics   20   26.0           Antiplatelets   55   71.4           Calcium Blockers   12   15.6           ARB   18   23.4                  
 
         [0060]    
       
         
               
             
               
               
               
               
             
               
               
               
               
               
               
               
             
               
               
               
               
               
             
               
               
               
               
               
               
               
             
           
               
                 TABLE 2 
               
             
             
               
                   
               
               
                   
               
               
                 Comparison of Pretreatment Clinical Variables and 
               
               
                 Risk Factors of Study Population by Patient Group 
               
             
          
           
               
                   
                 Responders 
                 NonResponders 
                   
               
               
                 Variables 
                 Number/(Percentage) 
                 Number/(Percentage) 
                 p Value 
               
               
                   
               
             
          
           
               
                 Total Study 
                   
                 44 
                 (100) 
                 33 
                 (100) 
                   
               
               
                 Number 
               
               
                 Gender 
                 Male 
                 37 
                 (84.1) 
                 25 
                 (75.8) 
                 0.361 
               
               
                   
                 Female 
                 7 
                 (15.9) 
                 8 
                 (24.2) 
               
             
          
           
               
                 Age/Groups 
                 Mean 
                 64.9 ± 8.9 
                 61.0 ± 8.4 
                 0.058 
               
               
                   
                 Range 
                 48-81 
                 42-80 
               
             
          
           
               
                   
                 Under 50 
                 2 
                 (4.5) 
                 2 
                 (6.1) 
                   
               
               
                   
                 50-59 
                 12 
                 (27.3) 
                 13 
                 (39.4) 
               
               
                   
                 60-69 
                 14 
                 (31.8) 
                 11 
                 (33.3) 
               
               
                   
                 70-79 
                 14 
                 (31.8) 
                 6 
                 (18.2) 
               
               
                   
                 80 and Over 
                 2 
                 (4.5) 
                 1 
                 (3.0) 
               
               
                 Coronary Risk 
                 Hypertension 
                 31 
                 (70.5) 
                 21 
                 (63.6) 
                 0.527 
               
               
                 Factors 
                 Hyperlipidemia 
                 41 
                 (93.2) 
                 27 
                 (81.8) 
                 0.160 
               
               
                   
                 Diabetes Mellitus 
                 11 
                 (25.0) 
                 9 
                 (27.3) 
                 0.822 
               
               
                   
                 Peripheral Vascular 
                 4 
                 (9.1) 
                 6 
                 (18.2) 
                 0.311 
               
               
                   
                 Disease 
               
               
                   
                 History of Congestive 
                 3 
                 (6.8) 
                 2 
                 (6.1) 
                 0.999 
               
               
                   
                 Heart Failure 
               
               
                   
                 Renal Insufficiency 
                 1 
                 (2.3) 
                 1 
                 (3.0) 
                 0.999 
               
               
                   
                 (Creatine &gt; 2.0 
               
               
                   
                 Previous Myocardial 
                 10 
                 (22.8) 
                 4 
                 (12.1) 
                 0.370 
               
               
                   
                 Stable Angina 
                 16 
                 (36.4) 
                 3 
                 (9.1) 
                 0.013 
               
               
                 Previous 
                 Prior Coronary Artery 
                 24 
                 (54.5) 
                 15 
                 (45.5) 
                 0.430 
               
               
                 Cardiovascular 
                 Bypass Grafting 
               
               
                 Intervention 
                 Prior Percutaneous 
                 10 
                 (22.7) 
                 7 
                 (9.1) 
                 0.999 
               
               
                   
                 Coronary Intervention 
               
               
                 Current 
                 Statins 
                 39 
                 (88.6) 
                 27 
                 (81.8) 
                 0.515 
               
               
                 Medications 
                 Nitrates 
                 16 
                 (36.4) 
                 7 
                 (21.2) 
                 0.151 
               
               
                   
                 Anticagulants 
                 2 
                 (4.5) 
                 1 
                 (3.0) 
                 0.999 
               
               
                   
                 Beta Blockers 
                 27 
                 (61.4) 
                 15 
                 (45.5) 
                 0.165 
               
               
                   
                 ACE Inhibitors 
                 15 
                 (34.1) 
                 13 
                 (39.4) 
                 0.632 
               
               
                   
                 Diuretics 
                 12 
                 (27.3) 
                 8 
                 (24.2) 
                 0.764 
               
               
                   
                 Antiplatelets 
                 32 
                 (72.7) 
                 23 
                 (69.7) 
                 0.771 
               
               
                   
                 Calcium Blockers 
                 5 
                 (11.4) 
                 7 
                 (21.2) 
                 0.238 
               
               
                   
                 ARB 
                 8 
                 (18.2) 
                 10 
                 (30.3) 
                 0.214 
               
               
                   
               
             
          
         
       
     
         [0061]    
       
         
               
             
               
               
               
               
             
           
               
                 TABLE 3 
               
             
             
               
                   
               
               
                   
               
               
                 Comparison of Initial and Ending 
               
               
                 CAC Scan Scores for Responders 
               
             
          
           
               
                 Variables 
                 Initial Score 
                 Ending Score 
                 p Value 
               
               
                   
               
               
                 Total Responder Number 
                 44 (100) 
                 44 (100) 
                   
               
               
                 Total Score 
                 2033.0 
                 1753.8 
                 0.001 
               
               
                 Left Coronary Artery 
                 89.5 
                 89.8 
                 0.972 
               
               
                 Left Anterior 
                 927.7 
                 788.4 
                 0.002 
               
               
                 Descending Coronary 
               
               
                 Artery 
               
               
                 Circumflex Coronary 
                 244.2 
                 211.1 
                 0.106 
               
               
                 Artery 
               
               
                 Right Coronary Artery 
                 771.0 
                 664.5 
                 0.002 
               
               
                   
               
             
          
         
       
     
         [0062]    
       
         
               
             
               
               
               
               
               
             
               
               
               
               
               
               
             
           
               
                 TABLE 4 
               
             
             
               
                   
               
               
                   
               
               
                 Comparison of Responder NB/CNP Antibody Levels (Units) and Antigen Levels (Units) 
               
             
          
           
               
                   
                 Beginning 
                 Two-Month 
                   
                   
               
               
                   
                 Value 
                 Value 
                 EndingValue 
               
               
                 Variables 
                 (Mean ± S.D.) 
                 (Mean ± S.D.) 
                 (Mean ± S.D.) 
                 p Value 
               
               
                   
               
             
          
           
               
                 NB/CNP 
                   
                 0.864 ± 0.35 
                 0.954 ± 0.485 
                 0.981 ± 0.50 
                   
               
               
                 Antibody 
               
               
                   
                 Beginning Value v. 
                   
                   
                   
                 0.269 
               
               
                   
                 Two Month Value 
               
               
                   
                 Beginning Value v. 
                   
                   
                   
                 0.300 
               
               
                   
                 Ending Value 
               
               
                   
                 Two-Month Value v. 
                   
                   
                   
                 0.799 
               
               
                   
                 Ending Value 
               
               
                 NB/CNP 
                   
                 2.045 ± 4.64 
                 2.067 ± 4.30  
                 4.012 ± 8.39 
               
               
                 Antigen 
               
               
                   
                 Beginning Value v. 
                   
                   
                   
                 0.982 
               
               
                   
                 Two Month Value 
               
               
                   
                 Beginning Value v. 
                   
                   
                   
                 0.206 
               
               
                   
                 Ending Value 
               
               
                   
                 Two-Month Value v. 
                   
                   
                   
                 0.219 
               
               
                   
                 EndingValue 
               
               
                   
               
             
          
         
       
     
         [0063]    
       
         
               
             
               
               
               
               
             
               
               
               
               
             
           
               
                 TABLE 5 
               
               
                   
               
               
                   
               
               
                 Comparison of Responders Beginning 
               
               
                 and Ending Cholesterol Levels 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                   
                 Beginning Value 
                 EndingValue 
                   
               
               
                   
                 (Mean ± S.D.) 
                 (Mean ± S.D.) 
               
               
                 Lipid Panel 
                 (mmol/L) 
                 (mmol/L) 
                 p Value 
               
               
                   
               
               
                 Total Cholesterol 
                 4.9 ± 1.2 
                 4.2 ± 0.88 
                 0.001 
               
               
                 Triglycerides 
                 2.5 ± 3.2 
                 1.9 ± 2.3 
                 0.006 
               
               
                 HDL Cholesterol 
                 1.2 ± 0.319 
                 1.4 ± 0.34 
                 0.001 
               
               
                 LDL Cholesterol 
                 2.6 ± 0.9 
                 2.1 ± 0.8 
                 0.001 
               
               
                   
               
             
          
           
               
                   
                 Beginning Value 
                 EndingValue 
                   
               
               
                   
                 (Mean ± S.D.) 
                 (Mean ± S.D.) 
               
               
                 Lipid Panel 
                 (mg/dL) 
                 (mg/dL) 
                 p Value 
               
               
                   
               
               
                 Total Cholesterol 
                 188.6 ± 47.4 
                 164.5 ± 33.8 
                 0.001 
               
               
                 Triglycerides 
                  232.0 ± 302.5 
                  179.4 ± 221.1 
                 0.006 
               
               
                 HDL Cholesterol 
                  47.4 ± 12.1 
                 52.25 ± 13.0 
                 0.001 
               
               
                 LDL Cholesterol 
                 101.8 ± 35.6 
                  81.3 ± 29.5 
                 0.001