CN1688301A - Dosage forms and related therapies - Google Patents

Abstract

This invention relates to novel doses, dosage formulations, and routes of administration of such doses and dose formulations, said dose and dose formulations containing one or more copper chelators, for example, one or more trientine active agents, including trientine analogues, trientine salts, trientine prodrugs, and trientine derivatives, useful in the treatment of diseases, disorders and conditions, including in indications where copper may play a role.

Description

Pharmaceutical dosage forms and related therapeutic uses

technical field

The present invention relates to dosages and formulations of therapeutic agents and their use in methods of treating, reversing or ameliorating diseases and/or conditions in mammals (referred to herein as "treatment"). Mammals treated with the dosages and formulations described herein include humans suffering or at risk of suffering microvascular and/or macrovascular damage, such as damage to cardiovascular tissue, in particular those suffering or at risk of having undesirable The development of copper levels includes humans, mammals, including the development of copper levels that cause or result in tissue damage including, but not limited to, vascular damage. Said treatment includes, but is not limited to, treatment to ameliorate and/or reverse, in whole or in part, damage caused by a disease, disorder, or condition characterized by damage to tissue and/or vasculature caused or mediated by copper Injury, and/or stem cell responses of normal tissues induced or mediated by copper. The present invention is by administering an active copper chelating substance such as one or more triethylenetetramine (trientine), a salt of triethylenetetramine, a prodrug of triethylenetetramine and the prodrug of said prodrug salts, analogs of triethylenetetramine and salts and prodrugs of said analogs, and/or active metabolites of triethylenetetramine and salts and prodrugs of such metabolites, including but not limited to N-acetyltriethyltetramine and its salts and prodrugs are especially useful in heart failure, macrovascular disease or injury, microvascular disease or injury, and/or intoxication (e.g., high blood pressure) diseases or injuries of tissues and/or organs (including diseases characterized by heart failure, cardiomyopathy, myocardial infarction and diseases of associated arteries and organs).

Background technique

The following description contains various information useful for understanding the present invention. It is not considered that any information described here belongs to the prior art or related content of the specification and claims of the present invention, nor is it considered that any publication or document specifically cited as a reference can be used as a patent for evaluating the specification and claims of the present invention. sex literature.

Diabetes is a chronic condition characterized by fasting high glucose and can develop into widespread premature atherosclerosis. Both morbidity and mortality in diabetic patients are increased due to the occurrence of cardiovascular disease, especially coronary artery disease. Vascular complications in diabetic patients can be divided into microvascular disease, which affects the retina, kidneys, and nerves, and macrovascular disease, which mainly affects coronary vessels, cerebral vessels, and peripheral arterial circulation.

Chronic hyperglycemia in diabetes is associated with long-term damage, dysfunction, and failure of various organs, especially the eyes, kidneys, nerves, heart, and blood vessels. Long-term complications of diabetes include retinopathy with a risk of blindness; can lead to renal renal disease; peripheral neuropathy, which can cause symptoms of foot ulcers, amputations, and charred joints; and autonomic neuropathy and sexual dysfunction, which can cause symptoms of the gastrointestinal, urinary, and cardiovascular systems.

Mechanisms thought to result in tissue damage due to chronic hyperglycemia include glycosylation of histones and other macromolecules and overproduction of polyol species derived from glucose. Diabetic patients have an increased incidence of atherosclerotic cardiovascular, peripheral vascular, and cerebrovascular disease. Hypertension, abnormal lipoprotein metabolism, and periodontal disease are also found in diabetic populations.

Hyperglycemia induces many changes in vascular tissue, likely contributing to the acceleration of atherosclerosis. Currently, in addition to nonenzymatic glycosylation of proteins and lipids, there are two other major mechanisms that comprise most of the pathological changes observed in diabetic animals and humans, namely oxidative stress and protein kinase C (PKC) activation. These mechanisms are not independent, for example, hyperglycemia-induced oxidative stress promotes the formation of AGEs and PKC activation, and both type 1 and type 2 diabetes independently induce the risk of coronary heart disease (CAD), stroke, and peripheral arterial disease factor. See Schwartz C.J., et al., "Pathogenesis of theatherosclerotic lesion. Implications for diabetes mellitus," Diabetes Care 15:1156-1167 (1992); Stamler J., et al., "Diabetes, other risk factors, and 12-yr cardiovascular mortality for men screened in the Multiple RiskFactor Intervention Trial." Diabetes Care 16: 434-444 (1993). Atherosclerosis accounts for virtually 80% of deaths among North American diabetic patients, equivalent to one-third of all deaths in the North American general population, and more than 75% of hospitalizations for diabetic complications are due to cardiovascular disease. See American Diabetes Association, "Consensus statement: role of cardiovascular risk factors in prevention and treatment of macrovascular disease in diabetes," Diabetes Care 16:72-78 (1993).

Declines in cardiac mortality in the general U.S. population have been attributed to reductions in cardiovascular risk factors and improvements in cardiac care. However, the age-dependent decrease in heart disease mortality observed in the non-diabetic population was not seen in patients with diabetes, and there was an increase in mortality among women with diabetes. See Gu K, et al., "Diabetes and decline in heart disease mortality in U.S. adults," JAMA 281: 1291-1297 (1999). It has also been reported that diabetic patients have more extensive coronary and cerebrovascular atherosclerosis than age- and sex-matched non-diabetic controls. See Robertson W.B., & Strong J.P., "Atherosclerosis in persons with hypertension and diabetes mellitus," Lab Invest 18:538-551 (1968). In addition, it has been reported that diabetic patients have more involved coronary vascular disease and more diffuse atherosclerotic lesions. See Waller, B.F., et al., "Status of the coronary arteries at necropsy in diabetes mellitus with onset after age 30 years. Analysis of 229 diabetic patients with and without clinical evidence of sub-coronary heart disease and comparisons to" J. l ed con, 183 : 498-506 (1980).

Diabetic patients with CAD who underwent cardiac catheterization, angioplasty, or coronary artery bypass surgery for acute myocardial infarction were also reported to have significantly more For severe proximal and distal CAD. See Granger C.B., et al., "Outcome of patients with diabetes mellitus and acute myocardial injury treated with thrombolytic agents. The Thrombolysis and Angioplasty in Myocardial Infarction (TAMI) Study Group," JAm Coll Cardiol 21:919-935; ., "Influence of diabetes mellitus on early and late outcome after percutaneous transluminal coronary angioplasty," Circulation 91: 979-989 (1995); Barzilay J.I., "Coronary artery disease and coronary disease by passent diabet >or=65 years [from the Coronary Artery Surgery Study (CASS) Registry]," Am J Cardiol 74:334-339 (1994)). Evidence from autopsy and vascular fluoroscopy also showed a marked increase in ulcerated plaques and thrombus formation in diabetic patients. See Davies M.J., et al., "Factors influencing the presence or absence of acute coronary artery thrombosis inudden ischemic death," Eur Heart J 10; 203-208 (1989); Silva J.A., et al., "Unstable angina. A comparison of angioscopic Findings between diabetic and nondiabetic patients," Circulation 92: 1731-1736 (1995).

CAD causes death in patients with type 2 diabetes regardless of the duration of diabetes. See Stamler I., et al., "Diabetes, other risk factors, and 12-yrcardiovascular mortality for men screened in the Multiple Risk Factor Intervention Trial," Diabetes Care 16:434-444 (1993); Donahue R.P., & Orchard T.J., in "Diabetes mellitus and macrovascular complications. Anepidemiological perspective," Diabetes Care 15:1141-1155 (1992). The increased risk of cardiovascular disease is particularly evident in the female population. See Barrett Connor E.L., et al., "Why is diabetes mellitus a stronger risk factor for fatal ischemic heart disease in women than in men? The Rancho Bemardo Study," JAMA 265:627-631 (1991). The occurrence of CAD does not limit the specific type of diabetes, which is common in both type 1 and type 2 diabetes, and the mortality rate of cardiovascular disease increases significantly after the age of 30. See Krolewski A.S., et al., "Magnitude and determinants of coronary artery disease in juvenile-onset, insulin-dependent diabetes mellitus," Am J Cardiol 59:750-75 5 (1987). The study reported that the incidence of CAD increased rapidly after the age of 40. At the age of 55, 35% of male and female patients with type 1 diabetes died of CAD, and the mortality rate of CAD was much higher than that of non-diabetic people of the same age.

Diabetic nephropathy in patients with type 1 diabetes also increases the incidence of CAD complications. Renal disease leads to exacerbated accumulation of AGEs in the systemic circulation and tissues, and similarly severe renal impairment. See Makita Z., et al., "Advanced glycosylation end products inpatients with diabetic nephropathy," N Engl J Med 325:836-842 (1991). Among diabetic patients who reached advanced nephropathy, overall mortality was also higher than in the non-diabetic population. Among all patients with diabetes, the relative age-specific death rate from myocardial infarction was 89 times that of the general population in the first year after diagnosis. See Geerlings W., et al., "Combined report on regular dialysis and transplantation in Europe, XXI," Nephrol Dial Transplant 64]: 5-29 (1991). It has also been reported that the most common cause of death in kidney transplanted diabetic patients is CAD, accounting for 40% of such patients' deaths. See Lemmers M.J., & Barry J.M., "Major role for arterial disease injury and mortality after kidney transplantation in diabetic recipients," Diabetes Care 14:295-301 (1991).

The degree and duration of hyperglycemia have been shown to be major factors in the pathogenesis of microvascular complications in type 2 diabetes. See The Diabetes Control and Complications Trial Research Group, "The effect of intensive treatment of diabetes on the development and progression of long-term complications ininsulin-dependent diabetes mellitus," N EngJ Med 329:977-986 (1993). However, the relationship between the severity of macrovascular complications and the degree or duration of diabetes is not well understood, and it has been reported that a general increase in the incidence of CAD in patients with newly diagnosed type 2 diabetes is evident. See Uusitupa M., et al., "Prevalence of coronary heart disease, left ventricular failure and hypertension in middle-aged, newly diagnosed type 2 (non-insulin dependent) diabetic subjects," Diabetologia 28: 22-27 (1985). It has also been reported that impaired glucose tolerance also contributes to an increased incidence of cardiovascular disease despite its reduction in hyperglycemia. See Fuller J.H., et al., "Coronary-heart-disease risk and impaired glucose tolerance. The Whitehall study," Lancet 1:1373-1376 (1980).

The rising incidence of diabetes is also a worldwide trend. The number of cases of type 2 diabetes reached 135,000,000 in 2000 and will exceed 300,000,000 in 2025. These increases are associated with increased life expectancy, increases in obesity, and poor socioeconomic status. See WHO's 1997 World Health Report. As a result, mortality from diabetes has increased in the last decade, while mortality from cardiovascular disease, stroke, and malignancy has remained constant or decreased. See US Centers for Health Research. Causes of death from type 2 diabetes included cardiovascular disease, 58%; cerebrovascular disease, 12%; renal disease, 3%; diabetic coma, 1%; and malignancy, 11%.

Diabetic heart disease is further characterized by more severe CAD in younger patients, a 4-fold increased incidence of heart failure, post-acute myocardial infarction and increased left ventricular hypertrophy and disproportion. See Struthers A.D., & Morris A.D., Lancet 359:1430-2 (2002). Patients with type 2 diabetes also showed a disproportionate increase in mortality in the first 24 hours after acute myocardial infarction. Emergency intervention can improve this risk. See Malmberg K., BrMed J 314:1512-5 (1997).

PCT Application No. PCT/NZ99/00161 (published as WO00/18392, 2000.04.06) relates to a method of treating mammals susceptible to induction and/or suffering from diabetes to reduce macrovascular and microvascular damage, comprising, in addition to controlling blood sugar In addition to the level of treatment, there is also the control of copper, for example, at least for a certain period of time. Its assay analysis method is described in PCT Application No. PCT/NZ99/00160 (published as WO00/18891, 2000.04.06). Various therapeutic agents are described in PCT/NZ99/00161. These include copper chelating agents.

Metals occur naturally in the body and many are essential for cells (eg Cu, Fe, Mn, Ni, Zn). However, all metals are toxic in high concentrations. One reason for metal toxicity is related to the ability of metals to cause oxidative stress, especially redox-active transition metals that accept or donate electrons (e.g., Fe2+/3+, Cu+/2±), causing free radical Increase causes damage (see "Evidence for the generation of hydroxyl radicals from a chromium (V) intermediate isolated from thereaction of chromate with glutathione," by Jones et al., Biochim. Biophys. Acta 286: 652-655 (1991); Li, Y . & Trush, M.A., "DNA damage resulting from the oxidation of hydroquinone by copper: role for a Cu(II)/Cu(I) redox cycle and reactive oxygen generation," Carcinogenes 7:1303-1311(1993)). Metals can displace other essential metals or enzymes, disrupting the function of these molecules and thus becoming toxic. Some metal ions (for example, Hg+ and Cu+) react very easily with sulfhydryl groups, affecting the structure and function of proteins.

Here, patients with type 2 diabetes or abnormal glucose metabolism are particularly prone to heart failure and other diseases of the arterial tree. It has been reported that more than 50% of patients with type 2 diabetes in Western countries die from the effects of cardiovascular disease. See Stamler, et al. Diabetes Care 16:434-44 (1993). It has also been reported that a low degree of glucose intolerance as determined by a glucose tolerance test (impaired glucose tolerance, or "IGT") is associated with an increased risk of sudden death. See Balkau, et al. Lancet 354:1968-9 (1999). Over longer periods of time, it is assumed to reflect an increased incidence of coronary atherosclerosis and myocardial infarction in diabetic patients. However, evidence suggests that diabetes can cause specific heart failure or cardiomyopathy in the absence of atherosclerotic coronary artery disease.

Cardiac function can usually be assessed by measuring ejection fraction. A normal left ventricle ejects at least 50% of its end-diastolic volume of blood with each pump. Patients with systolic heart failure typically eject less than 30% of the left ventricle with a compensatory increase in end-diastolic volume. Hemodynamic studies in diabetic patients without overt congestive heart failure have observed normal left ventricular systolic function (LV ejection fraction) but abnormal diastolic function, suggesting impaired left ventricular diastole or perfusion. See Regan, et al. J Gun. Invest. 60:885-99 (1977). In a recent study, 60% of patients with type 2 diabetes without clinically significant evidence of heart failure had diastolic filling abnormalities, as assessed by echocardiography. See Poirier, et al. Diabetes Care 24:5-10 (2001). Diagnosis is accomplished, for example, by non-invasive assay methods. In patients without mitral stenosis, mitral diastolic flow measured by Doppler echocardiography can be used directly to measure left ventricular perfusion. The most common assay is the AlE ratio. Normal early diastolic filling is rapid and is characterized by E-wave velocities of approximately 1 m/s. Late diastolic filling by atrial contraction is only a minor component, with A-wave velocities of approximately 0.5 m/s. This gives a normal AIE ratio of about 0.5. For diastolic dysfunction, early diastolic filling is impaired, atrial contraction provides a compensatory increase, and the AlE ratio increases to more than 2.0.

Treatment of diabetic cardiomyopathy, whether reversing or improving, is difficult and options are limited. Tight control of blood sugar levels can prevent or reverse heart failure, but only in the early stages of ventricular failure. Angiotensin-converting enzyme inhibitors such as captopril can improve survival in heart failure, especially in patients with severe systolic heart failure and the lowest ejection index. However, there are many treatments that are not recommended for diabetic cardiomyopathy. For example, inotropic drugs can improve the contraction of a failing heart. However, hearts with isolated diastolic dysfunction already contract normally, and inotropic drugs are believed to increase the likelihood of arrhythmia. In addition, vasodilators that reduce afterload and improve ventricular emptying may also be used, since ejection index and end-diastolic volume are already normalized. Decreased afterload may worsen cardiac function by creating outflow tract obstruction.

Diuretics are the main drugs in the treatment of heart failure, which achieve their effect by controlling the retention of salt and water and reducing filling pressure. However, the use of diuretics is contraindicated in the treatment of diastolic dysfunction in which compromised cardiac pump function relies on high filling pressures to maintain cardiac output. Venodilators, such as nitrates, are very effective in the treatment of systolic heart failure by reducing preload and filling pressures, but are understood to be less well tolerated in patients with diastolic heart failure. Ejection index and end-systolic volume are usually normal, and any decrease in preload results in a marked decrease in cardiac output. Finally, the use of beta-blockers in heart failure is also relevant because they may worsen pump function. It also relates to the administration of beta-blockers to diabetic patients, who are treated with sulfonylurea drugs and insulin, which may cause severe hypoglycemic conditions.

Therefore, it can be understood that the mechanisms according to the different diseases of the heart, macrovascular system, microvascular system and long-term complications of diabetes, including those related to heart diseases and long-term complications, are very complex and have been studied for a long time and are not clearly disclosed, The safety and effectiveness of its treatment is also unknown. There is therefore a need for such treatment, as described in the specification and claims of the present invention.

It is also understood that there is a need for a pharmaceutical composition capable of addressing damage caused by a disease, disorder or condition of the cardiovascular tree (including the heart) and corresponding organs (e.g., retina, kidney, nerves, etc.) With respect to, for example, elevated or unwanted levels of copper, such as elevated levels of non-intracellular free copper. The methods of treatment described herein also provide low dose controlled release and/or low dose extended release compositions for reversing and/or ameliorating structural damage in a subject with or without diabetes, which Having copper levels that can be lowered to treat heart, macrovasculature, microvasculature disease and/or long-term diabetic complications, and including damage to cardiac structures. Cardiac structural damage including, but not limited to, atrophy, loss of cardiomyocytes, expansion of the extracellular space and increased deposition of extracellular matrix (and consequential consequences) and/or damage to at least the medial layer (muscle layer) and the intimal layer Structural damage to coronary arteries (and consequences), systolic function, diastolic function, contractility, recoil characteristics, and ejection index of each layer (endothelial layer).

Diseases, disorders and conditions associated with the cardiovascular tree and/or dependent organs can be treated with the methods and compositions described herein, including, for example, the treatment of any one or more of the following diseases: (1) Cardiomyopathy (cardiomyopathy or myocarditis) such as idiopathic cardiomyopathy, metabolic cardiomyopathy including diabetic cardiomyopathy, alcoholic cardiomyopathy, drug-induced cardiomyopathy, ischemic cardiomyopathy, and hypertensive cardiomyopathy; (2 ) atherosclerotic disease of major vessels (macrovascular system disease) such as the aorta, coronary, carotid, cerebral, renal, iliac, femoral, and popliteal arteries; (3) poisoning, drug-induced, and Metabolic (including hypertension and/or diabetes mellitus of small vessels (microvasculature) such as retinal arterioles, glomerulus arterioles, neurovascular arterioles, cardiac arterioles, and those associated with the eyes, kidneys, heart, and central and peripheral nerves vessels associated with systemic capillary beds; and (4) plaque rupture in areas of atherosclerosis in major vessels such as the aorta, coronary, carotid, cerebral, renal, iliac, femoral, and popliteal arteries.

Summary of the invention

The present invention is based in part on novel dosages and formulations for use as treatments for effective free copper reduction, eg, treatment or prevention of macrovascular, microvascular, and/or toxic/metabolic diseases, and tissue repair processes as described herein. This was independent of the subjects' glucose metabolism, and whether fructosamine oxidase was involved in these disorders. The invention also relates to dosage forms for the treatment of cardiovascular diseases associated with the accumulation of redox-active transition metal ions in diabetic patients.

Under physiological conditions, the damage to the target organ is sensed by distant stem cells, which migrate to the damaged site and help repair the structure and function through stem cell differentiation. Therapeutic doses and formulations described here also slow the accumulation of redox transition metals, particularly copper, in the myocardium or vascular tissue of diabetic patients, and without wishing to be bound by theory, it is believed that this accumulation is concomitantly influenced by stem cell migration inhibit the regeneration of normal tissues. The suppression of normal biological behavior of undifferentiated cells by elevated copper levels in tissues is independent of the diabetic condition, although this is common in diabetic mammals, including humans.

Impaired glucose tolerance, the symptoms and/or suppressed normal stem cell responses seen in diabetes mellitus, can cause impairment of normal tissue responses, and this can be ameliorated by lowering copper levels using the therapeutic doses and formulations described herein, including the following content:

1. Heart failure. Significant regeneration of cardiac tissue occurs in the days following heart transplant surgery. The likely mechanism is the migration of stem cells outside the heart, and their subsequent differentiation to form various cardiac cells, including cardiomyocytes, endothelial cells, and coronary vascular cells. We have shown that accumulation of copper in cardiac tissue may severely impair these regenerative responses and may be used, for example, in acute intravenous therapy with copper chelators for the treatment of heart failure including, but not limited to, diabetic heart failure.

2. Acute myocardial infarction (AMI). AMI is often accompanied by hyperplasia of cells in the ventricular myocardium, such as occurs in diabetes mellitus as described. Elevated levels of redox transition metals suppress normal stem cell responses, leading to structural impairment and functional repair of damaged tissues. The mechanism of impaired cardiac function in, for example, diabetic patients is believed to be the toxic effect of transition metal accumulation on tissue dynamics, the impairment of tissue regeneration resulting from the normal stem cell response that regulates physiological tissue regeneration by inhibiting migration from the outside to the damaged tissue. Treatment of AMI, for example in diabetic patients as described herein, is improved by acute (parenteral administration if necessary) or chronic administration of the copper chelators.

3. Wound healing and ulcers. The repair process of normal tissue requires the intervention of active stem cells, which can affect the repair of various layers of blood vessels. Accumulation of transition metals (especially copper) in vascular tissue causes tissue damage characteristic of diabetes, including wound repair from surgery or trauma, and ulcer expansion and poor prognosis. Treatment of diabetes with copper chelators prior to surgery or during tissue injury as described herein may also advantageously employ the dosages and formulations described herein. Surgery in diabetic patients has better outcomes if excess transition metals are removed from blood vessels prior to surgery. This can be accomplished in an acute (parenterally administered therapy) and/or chronic manner (oral administered therapy) prior to actual surgery.

4. Tissue damage caused by infection. The normal tissue repair process after infection requires the intervention of active stem cells that can migrate to damaged tissue to regenerate and repair it, for example, the regeneration and repair of the various layers of blood vessels. Repair of these tissue lesions can be impaired by suppressed stem cell responses, as caused by the accumulation of redox transition metals (particularly copper) in tissues, for example, in blood vessel walls. In diabetic patients, repair of tissue damage, including, for example, repair after infection can be improved by using the therapeutic dosages and formulations described herein.

5. Diabetic kidney injury. The treatment of diabetes and other conditions with renal failure by giving the copper chelating agent in the dose and dosage form described in the present invention will improve the regeneration of the organ through the recovery of normal tissue healing, which is achieved by making stem cells normal Migration and differentiation are achieved.

However, even in non-diabetic mammals and mammals without abnormal glucose mechanisms, reduced extracellular copper levels are beneficial, and lower levels of copper lead to attenuation of copper-mediated tissue damage, and/or through Improves tissue repair by restoring normal tissue stem cell responses.

In the studies described here using the rat model of streptozotocin-induced diabetes (STZ), a high incidence of tissue damage in both cardiac and coronary tissue was found in severely diabetic animals and extrapolated to humans. Condition. In one aspect, the invention describes a method of reducing free copper levels in a subject with or without diabetes, particularly a patient without Wilson's disease, who has copper levels capable of being reduced by administering an effective amount of obtained from agents capable of reducing copper levels in patients.

Preferred copper chelators are triethylenetetramine, including triethylenetetramine acid addition salts, and active metabolites, including, for example, N-acetyltriethylenetetramine and analogs, derivatives and prodrugs thereof. Alternative names for triethylenetetramine include N,N'-bis(2-aminoethyl)-1,2-ethanedi-amine; triethylenetetramine ("TETA"); Amino-3,6-diazaoctane; 3,6-diazaoctane-1,8-diamine; 1,4,7,10-tetraazadecane; trientine; TECZA; and triene. In a specific embodiment, triethylenetetramine is non-basic (eg, as an acid addition salt).

In another embodiment, triethylenetetramine may be modified, i.e., may be an analog or derivative of triethylenetetramine (or an analog of a copper-chelated triethylenetetramine metabolite or derivatives, such as N-acetyltriethyltetramine). Derivatives of triethylenetetramine or triethylenetetramine salts or analogs thereof include those modified with polyethylene glycol (PEG). The structure of PEG is HO-(- _CH2 - _CH2 -O-) _n -H. It can be a linear or branched neutral polyether and can have different molecular weights. Analogs of triethylenetetramine include compounds in which one or more NH groups in triethylenetetramine are replaced by sulfur atoms. Other analogs include compounds in which triethylenetetramine has been modified to append one or more additional _-CH2 groups. The chemical formula of triethylenetetramine is _NH2 - _CH2 - _CH2 -NH- _CH2 - _CH2 -NH- _CH2 - _CH2 - _NH2 . Its empirical formula is C ₆ N ₄ H ₁₈ . Examples of triethylenetetramine analogs include:

1. SH-CH ₂ -CH ₂ -NH-CH ₂ -CH ₂ -NH-CH ₂ -CH ₂ -NH ₂ ,

2. SH-CH ₂ -CH ₂ -S-CH ₂ -CH ₂ -NH-CH ₂ -CH ₂ -NH ₂ ,

3. NH ₂ -CH ₂ -CH ₂ -NH-CH ₂ -CH ₂ -S-CH ₂ -CH ₂ -SH,

4. NH ₂ -CH ₂ -CH ₂ -S-CH ₂ -CH ₂ -S-CH ₂ -CH ₂ -SH,

5. SH- _CH2 - _CH2 -S- _CH2 - _CH2 -S- _CH2 - _CH2 -SH,

6. _{NH 2} -CH ₂ -CH ₂ -NH-CH ₂ -CH ₂ -CH ₂ -NH-CH ₂ -CH ₂ -NH ₂ ,

7. SH-CH ₂ -CH ₂ -NH-CH ₂ -CH ₂ -CH ₂ -NH-CH ₂ -CH ₂ -NH ₂ ,

8. SH-CH ₂ -CH ₂ -S-CH ₂ -CH ₂ -CH ₂ -NH-CH ₂ -CH ₂ -NH ₂ ,

9. NH ₂ -CH ₂ -CH ₂ -NH-CH ₂ -CH ₂ -CH ₂ -S-CH ₂ -CH ₂ -SH,

10. NH ₂ -CH ₂ -CH ₂ -S-CH ₂ -CH ₂ -CH ₂ -S-CH ₂ -CH ₂ -SH,

11. SH- _CH2 - _CH2 -S- _CH2 - _CH2 - _CH2 -S- _CH2 - _CH2 -SH,

12. Wait.

One or more hydroxyl groups may also be substituted for one or more amine groups, resulting in triethylenetetramine analogs (one or more nitrogen atoms may or may not be replaced by one or more sulfur atoms). Other analogs include acyclic and cyclic analogs represented by the following general formulas I and II.

In another embodiment, triethylenetetramine is delivered as a prodrug of triethylenetetramine or a copper chelated metabolite of triethylenetetramine.

Salts of triethylenetetramine (which may optionally be prodrugs of triethylenetetramine or salts of copper chelated metabolites of triethylenetetramine) include in one embodiment acid addition salts, as appropriate Salts of inorganic or organic acids. Salts of triethylenetetramine (acid addition salts, such as triethylenetetramine dihydrochloride) act as copper chelating agents, capable of eliminating copper from the body by forming stable soluble complexes with copper that are readily excreted by the kidneys .

Another aspect of the present invention is to provide a method of (1) ameliorating or reversing, in whole or in part, at least one or more structural impairments (e.g., atrophy, muscle cell loss, extracellular space expansion, and/or Increased deposition of extracellular matrix (and its consequences), and/or (2) total or partial improvement in one or more of systolic function, diastolic function, contractility, recoil characteristics, and ejection fraction (e.g., by ultrasound, MRI or other imaging techniques), and/or (3) fully or partially ameliorating or reversing diseases (and their consequences) of myocardial, macrovascular disease, microvascular disease, and/or atherosclerotic plaque rupture injury of major vessels, etc. The damage of the disease, and/or (4) fully or partially improve or reverse the damage caused by diabetic nephropathy, diabetic nephropathy, copper accumulation in the kidney and/or renal artery damage. The method may include:

(i) diagnosing that the mammal may suffer from an impairment that can be at least partially ameliorated and/or reversed, and

(ii) administering to a mammal a composition of triethylenetetramine active agents described herein.

In a specific embodiment, the composition is in a dosage form capable of providing a subject with a lower effective dose and less pulsile exposure than the "QID" of the Wilson's disease treatment regimen.

Another aspect of the invention includes methods of ameliorating or reversing, in whole or in part, in (I) a diabetic human or other diabetic animal, or (II) having the ability to reduce ("subject") one or Various atrophy, loss of myocytes, expansion of the extracellular space, and/or increased deposition of extracellular matrix (and its consequences) and/or copper levels of coronary artery structural damage, including medial lesions, in humans or other animals (muscular layer) and intimal layer (endothelial layer) of coronary artery structural damage (and the consequences). The method comprises or comprises administering to the subject or self-administering slow or sustained release of a dosage form providing sufficient copper chelation, the dosage form comprising a triethylenetetramine active agent, at least one triethylenetetramine Salts, prodrugs or salts of prodrugs, analogs or prodrugs or salts of analogs and/or metabolites or salts or prodrugs of triethylenetetramine, including but not limited to N-acetyltriethylenetetramine and its salts and prodrugs ("triethylenetetramine active agents").

In a specific embodiment said subject is treated as a susceptible population prior to treatment.

Another aspect of the present invention is a method comprising full or partial amelioration or reversal of one or more of systolic dysfunction, diastolic dysfunction, contractility, lack of desired recoil characteristics and/or or required as a function of ejection index (which can be determined by ultrasonography, MRI, or other imaging techniques), myocardial disease, macrovascular disease, microvascular disease, and microvascular disease and atherosclerotic plaque rupture of major vessels (and resulting consequences), the subject may be (I) a diabetic subject, or (II) a subject capable of reducing copper levels, the method comprising providing the subject with effective therapeutically effective, e.g., sufficient chelation A low dose, slow and/or controlled release dosage form comprising one or more copper chelating agents, such as one or more triethylenetetramine active agents, in an amount of copper chelating.

Diseases, disorders and conditions commonly referred to as being treated with the tissues and procedures of the present invention include, but are not limited to, one or more of the following: diabetic cardiomyopathy, diabetic acute coronary syndrome (such as myocardial infarction) MI), diabetic hypertensive cardiomyopathy, acute coronary syndrome associated with impaired glucose tolerance (IGT), acute coronary syndrome associated with impaired fasting glucose (IFG), hypertensive cardiomyopathy associated with IGT , hypertensive cardiomyopathy associated with IFG, ischemic cardiomyopathy associated with IGT, ischemic cardiomyopathy associated with IFG, ischemic cardiomyopathy associated with coronary artery disease (CHD), and glucose Acute coronary syndrome unrelated to metabolism, hypertensive cardiomyopathy unrelated to glucose metabolism, ischemic cardiomyopathy unrelated to glucose metabolism (regardless of ischemic cardiomyopathy related to coronary artery sclerosis), and one or more diseases of the treeing vessels, including aorta, carotid, cerebrovascular, coronary, renal, retinal, vasa vasa, iliac vessels, femoral vessels, popliteal vessels, arteriolar tree, and capillaries bed disease.

A further aspect of the invention comprises the use of at least one triethylenetetramine active agent in dosage form with other suitable materials for the manufacture of a drug for the complete or partial amelioration or reversal of (I) suffering from diabetes or (II) capable of lowering copper levels Subjects with one or more of systolic function, diastolic function, contractility, recoil properties, and ejection index (which can be determined by ultrasound, MRI, or other imaging techniques), and/or one or more of at least Due in part to damage caused by diabetic nephropathy, diabetic nephropathy and/or accumulation of copper in the kidney, and/or damage to the renal arteries, and/or selected from one or more of atrophy, loss of cardiomyocytes, expansion of the extracellular space Damage to cardiac structures with increased deposition of extracellular matrix, and/or damage to selected coronary arteries that is associated or unrelated to damage to at least the middle layer (muscle layer) and the intimal layer (endothelial layer) Slow-release therapy for damage dosage form.

Another aspect of the invention provides a method of treating a subject having one or more of the symptoms described herein, the method comprising parenterally administering a therapeutically effective amount of a copper chelated Compositions of mixtures, wherein the therapeutically effective dose refers to about 5 mg-1100 mg per dose and/or per day.

In one embodiment the copper chelator is a triethylenetetramine active agent. Triethylenetetramine active agents include salts of triethylenetetramine, triethylenetetramine prodrugs or salts of such prodrugs, triethylenetetramine analogs or salts or prodrugs of such analogs, and/ Or at least one metabolite of triethylenetetramine or a salt or prodrug of the metabolite, including but not limited to N-acetyltriethylenetetramine and salts and prodrugs of N-acetyltriethylenetetramine. Triethylenetetramine active agents also include analogs of formulas I and II.

In a specific embodiment, other therapeutically effective amounts of triethylenetetramine active agents include, but are not limited to, triethylenetetramine, triethylenetetramine salts, triethylenetetramine analogs of formulas I and II, Etc., its effective amount includes 10mg-1100mg, 10mg-1000mg, 10mg-900mg, 20mg-800mg, 30mg-700mg, 40mg-600mg, 50mg-500mg, 50mg-450mg, from 50-100mg to about 400mg, from 50- 100mg to about 300mg, 110-290mg, 120-280mg, 130-270mg, 140-260mg, 150-250mg, 160-240mg, 170-230mg, 180-220mg, 190-210mg, and/or any other amount within the range.

The composition may include, for example, solutions, suspensions, and emulsions according to the efficiency requirements of parenteral administration, and may be administered by subcutaneous, intravenous, intramuscular, intradermal, intrasternal injection or infusion techniques.

The formulation may further comprise any one or more of the following: Buffering agents such as acetate, phosphate, citrate or glutamate buffers to obtain a final pH of the formulation in the range of approximately 5.0-9.5 , a carbohydrate or polyol regulator, which may be selected from antimicrobial preservatives such as m-cresol, benzyl alcohol, methyl, ethyl, propyl and butyl parabens (parabens) and phenol, and stabilizer.

Use enough water for injection to obtain the solution of the desired concentration. Sodium chloride and other excipients can also be used if desired. But these excipients must be able to maintain the overall stability of the triethylenetetramine active agent.

Formulations of the invention should be substantially isotonic. Isotonic solutions can be defined as containing certain concentrations of electrolytes, non-electrolytes, or combinations thereof such that introduction into mammalian tissues creates equal osmotic pressure. "Essentially isotonic" means isotonic within ±20%, preferably within ±10%. The formulated product may be contained in a container, such as typically a vial, cartridge, prefilled syringe or disposable pen.

Another aspect of the present invention is to provide a parenteral composition comprising a therapeutically effective amount of a copper chelator, which can be administered to a subject having one or more of the symptoms described below.

These conditions include diabetic cardiomyopathy, diabetic acute coronary syndrome (eg, myocardial infarction-MI), diabetic hypertensive cardiomyopathy, acute coronary syndrome associated with impaired glucose tolerance (IGT), and fasting glucose-impaired (IFG) associated acute coronary syndrome, hypertensive cardiomyopathy associated with impaired glucose tolerance (IGT), hypertensive cardiomyopathy associated with fasting glucose impairment (IFG), associated with impaired glucose tolerance (IGT) ischemic cardiomyopathy associated with fasting glucose impairment (IFG), ischemic cardiomyopathy associated with coronary heart disease (CHD), myocardial disorders (cardiomyopathy or myocarditis ) includes idiopathic cardiomyopathy, metabolic cardiomyopathy (including diabetic cardiomyopathy, alcoholic cardiomyopathy, drug-induced cardiomyopathy, ischemic cardiomyopathy, and hypertensive cardiomyopathy), and Acute coronary syndrome, hypertensive cardiomyopathy unrelated to abnormal glucose metabolism, ischemic cardiomyopathy unrelated to abnormal glucose metabolism (regardless of whether ischemic cardiomyopathy is related to coronary heart disease or coronary atherosclerotic heart disease) and Disease of one or more of the vascular trees, including the aorta, carotid and cerebral arteries, coronary arteries, renal arteries, retinal arteries, iliac arteries, femoral arteries, popliteal arteries, vasa vasa, arteriolar trees, and capillaries Atherosclerotic disease of major blood vessels (macrovascular disease) such as aortic, coronary, carotid, cerebral, renal, iliac, femoral and popliteal arteries, including but Structural damage to the heart not limited to atrophy, loss of myocytes, expansion of the extracellular space and increased deposition of extracellular matrix (and its consequences) and/or selected from at least the middle layer (muscle layer) and/or the intimal layer ( endothelial layer) damage (and consequences) coronary artery structural damage, atherosclerotic plaque rupture of major vessels such as aorta, coronary artery, carotid artery, cerebral artery, renal artery, iliac artery, femoral artery and popliteal artery disorders, systolic dysfunction, diastolic dysfunction, systolic disturbances, recoil characteristics and ejection index, toxicity, drug-induced and metabolic abnormalities including hypertension and/or diabetic disorders of small vessels (microvascular disease ) such as retinal arterioles, glomerulus arterioles, vasa vasa, cardiac arterioles and associated capillary beds of the eye, kidney, heart and central and peripheral nervous systems.

In one embodiment the copper chelator is a triethylenetetramine active agent. Triethylenetetramine active agents include salts of triethylenetetramine, triethylenetetramine prodrugs or salts of such prodrugs, triethylenetetramine analogs or salts or prodrugs of such analogs and and/or at least one active metabolite of triethylenetetramine or a salt or prodrug of such metabolite, including but not limited to N-acetyltriethylenetetramine and salts and prodrugs of N-acetyltriethylenetetramine body drugs.

A therapeutically effective amount of a copper chelator, e.g., one or more triethylenetetramine active agents, including but not limited to triethylenetetramine, triethylenetetramine salts, triethylenetetramines of formulas I and II Amine analogs and the like, the effective amount is about 5mg-1200mg per day. Other therapeutically effective dosage ranges include 10 mg-1100 mg, 10 mg-1000 mg, 10 mg-900 mg, 20 mg-800 mg, 30 mg-700 mg, 40 mg-600 mg, 50 mg-500 mg, 50 mg-450 mg, from 50-100 mg to about 400 mg, from 50- 100mg to about 300mg, 110-290mg, 120-280mg, 130-270mg, 140-260mg, 150-250mg, 160-240mg, 170-230mg, 180-220mg, 190-210mg, and/or any other amount within the range.

According to the efficiency requirements of parenteral administration, the composition may include, for example, solutions, suspensions, emulsions, and may be administered by subcutaneous, intravenous, intramuscular, intradermal, intrasternal injection or infusion techniques.

The formulation may further comprise any one or more of the following: Buffering agents such as acetate, phosphate, citrate or glutamate buffers to obtain a final pH of the formulation in the range of approximately 5.0-9.5 , a carbohydrate or polyol regulator, an antimicrobial preservative selected from m-cresol, benzyl alcohol, methyl, ethyl, propyl and butyl parabens and phenol, and a stabilizer.

Use enough water for injection to obtain the solution of the desired concentration. Sodium chloride and other excipients can also be used if desired. But these excipients must be able to maintain the overall stability of the triethylenetetramine active agent.

Formulations of the invention should be substantially isotonic. Isotonic solutions can be defined as containing certain concentrations of electrolytes, non-electrolytes, or combinations thereof such that introduction into mammalian tissues creates equal osmotic pressure. "Essentially isotonic" means isotonic within ±20%, preferably within ±10%. The formulated product may be contained in a container, such as typically a vial, cartridge, prefilled syringe or disposable pen.

Another aspect of the present invention is to provide a therapeutically effective amount of copper chelating agent in the preparation of the treatment of a subject with one or more of the following symptoms of the medicament: diabetic cardiomyopathy, diabetic acute coronary syndrome ( such as myocardial infarction-MI), diabetic hypertensive cardiomyopathy, acute coronary syndrome associated with impaired glucose tolerance (IGT), acute coronary syndrome associated with impaired glucose tolerance (IFG), associated with impaired glucose tolerance ( IGT-related hypertensive cardiomyopathy, fasting glucose-impaired (IFG)-related hypertensive cardiomyopathy, impaired glucose tolerance (IGT)-related ischemic cardiomyopathy, fasting-glucose-impaired (IFG)-related ischemic cardiomyopathy associated with coronary artery disease coronary heart disease (CHD), myocardial disorders (cardiomyopathy or myocarditis) including idiopathic cardiomyopathy, metabolic cardiomyopathy (including diabetic cardiomyopathy, alcoholic cardiomyopathy, drug-induced cardiomyopathy, ischemic cardiomyopathy, and hypertensive cardiomyopathy), acute coronary syndrome unrelated to abnormal glucose metabolism, hypertensive cardiomyopathy unrelated to abnormal glucose metabolism Ischemic cardiomyopathy unrelated to abnormal glucose metabolism (regardless of whether ischemic cardiomyopathy is related to coronary artery disease or coronary heart disease) and disease of one or more vascular trees, including aorta, carotid artery Arteries of major vessels (macrovascular disease) and disorders including arterial cerebral arteries, coronary arteries, renal arteries, retinal arteries, iliac arteries, femoral arteries, popliteal arteries, vasa vasa, arteriolar trees, and capillary beds Atherosclerotic disease such as disorders of the aorta, coronary, carotid, cerebral, renal, iliac, femoral, and popliteal arteries, including but not limited to atrophy, loss of myocytes, expansion of the extracellular space, and extracellular matrix Structural damage to the heart and/or structural damage to coronary arteries selected from damage (and consequences) to at least the middle layer (muscular layer) and/or the intimal layer (endothelial layer) of increased deposition of , plaque rupture of atherosclerotic lesions of major vessels such as aortic, coronary, carotid, cerebral, renal, iliac, femoral and popliteal arteries, systolic dysfunction, diastolic dysfunction, Systolic disturbances, recoil characteristics and ejection index, toxicity, drug-induced and metabolic abnormalities including hypertension and/or diabetic disorders of small vessels (microvascular disease) such as retinal arterioles, glomerular arterioles, vasa vasa, Cardiac arterioles and associated capillary beds of the eye, kidneys, heart, and central and peripheral nervous systems.

In one embodiment the copper chelator is a triethylenetetramine active agent. Triethylenetetramine active agents include salts of triethylenetetramine, triethylenetetramine prodrugs or salts of such prodrugs, triethylenetetramine analogs or salts or prodrugs of such analogs and and/or at least one active metabolite of triethylenetetramine or a salt or prodrug of such metabolite, including but not limited to N-acetyltriethylenetetramine and salts and prodrugs of N-acetyltriethylenetetramine body drugs.

A therapeutically effective amount of a copper chelator, e.g., a triethylenetetramine active agent, including but not limited to triethylenetetramine, triethylenetetramine salts, triethylenetetramine analogs of formulas I and II, and the like , an effective amount of which is about 5 mg-1200 mg per day. Other therapeutically effective dosage ranges include 10 mg-1100 mg, 10 mg-1000 mg, 10 mg-900 mg, 20 mg-800 mg, 30 mg-700 mg, 40 mg-600 mg, 50 mg-500 mg, 50 mg-450 mg, from 50-100 mg to about 400 mg, from 50- 100mg to about 300mg, 110-290mg, 120-280mg, 130-270mg, 140-260mg, 150-250mg, 160-240mg, 170-230mg, 180-220mg, 190-210mg, and/or any other amount within the range.

The composition may include, for example, solutions, suspensions, and emulsions according to the efficiency requirements of parenteral administration, and may be administered by subcutaneous, intravenous, intramuscular, intradermal, intrasternal injection or infusion techniques.

The formulation may further comprise any one or more of the following: Buffering agents such as acetate, phosphate, citrate or glutamate buffers to obtain a final pH of the formulation in the range of approximately 5.0-9.5 , a carbohydrate or polyol regulator, an antimicrobial preservative selected from m-cresol, benzyl alcohol, methyl, ethyl, propyl and butyl parabens and phenol, and a stabilizer.

Use enough water for injection to obtain the solution of the desired concentration. Sodium chloride and other excipients can also be used if desired. But these excipients must be able to maintain the overall stability of the triethylenetetramine active agent.

The formulations of the invention should be isotonic. An isotonic solution can be defined as containing a concentration of electrolytes, non-electrolytes, or a combination thereof such that introduction into mammalian tissues creates equal osmotic pressure. "Essentially isotonic" means isotonic within ±20%, preferably within ±10%. The formulated product may be contained in a container, such as typically a vial, cartridge, prefilled syringe or disposable pen.

Here, parenteral administration includes, but is not limited to, one or more of the following routes of administration: subcutaneous administration, intravenous administration, intramuscular administration, intraperitoneal administration, intrasternal administration, intraarticular administration medicament or intrasternal injection or infusion techniques (e.g., as sterile injectable aqueous or non-aqueous solutions or suspensions); nasal administration, such as inhalation spray; topical administration, such as in the form of a cream or ointment; or vaginal administration .

Treatment can be monitored by regular 24-hour urine copper analysis. Urine must be collected in copper-free glassware. A patient is considered to be in an ideal negative copper balance if the amount of copper in urine collected over a 24-hour period is between 0.5-1.0 mg.

One aspect of the invention is to provide a method of treating a subject having, for example, one or more of the symptoms described herein, comprising parenterally administering a composition comprising a therapeutically effective amount of copper For the chelating agent, the therapeutically effective amount for parenteral administration is in the range of about 0.1 mg/kg-40 mg/kg per dose, based on the weight of the subject.

In another specific embodiment, a therapeutically effective amount of a copper chelator, e.g., a triethylenetetramine active agent, including but not limited to triethylenetetramine, triethylenetetramine salts, triethylenetetramines of general formulas I and II Ethylenetetramine analogues and the like, the therapeutically effective amount is about 5 mg-1200 mg per day. Other therapeutically effective dosage ranges include 10 mg-1100 mg, 10 mg-1000 mg, 10 mg-900 mg, 20 mg-800 mg, 30 mg-700 mg, 40 mg-600 mg, 50 mg-500 mg, 50 mg-450 mg, from 50-100 mg to about 400 mg, from 50- 100 mg to about 300 mg, 110-290 mg, 120-280 mg, 130-270 mg, 140-260 mg, 150-250 mg, 160-240 mg, 170-230 mg, 180-220 mg, 190-210 mg, and/or any other stated range .

The composition may include, for example, solutions, suspensions, and emulsions according to the efficiency requirements of parenteral administration, and may be administered by subcutaneous, intravenous, intramuscular, intradermal, intrasternal injection or infusion techniques.

The formulation may further comprise any one or more of the following: Buffering agents such as acetate, phosphate, citrate or glutamate buffers to obtain a final pH of the formulation in the range of approximately 5.0-9.5 , a carbohydrate or polyol regulator, an antimicrobial preservative selected from m-cresol, benzyl alcohol, methyl, ethyl, propyl and butyl parabens and phenol, and a stabilizer.

Use enough water for injection to obtain the solution of the desired concentration. Sodium chloride and other excipients can also be used if desired. But these excipients must be able to maintain the overall stability of the triethylenetetramine active agent.

The formulations of the invention should be isotonic. An isotonic solution can be defined as containing a concentration of electrolytes, non-electrolytes, or a combination thereof such that introduction into mammalian tissues creates equal osmotic pressure. "Essentially isotonic" means isotonic within ±20%, preferably within ±10%. The formulated product may be contained in a container, such as typically a vial, cartridge, prefilled syringe or disposable pen.

Further aspects of the invention include transdermal patches, pads, packs or bandages ("adhesives") capable of being adhesively or otherwise bonded to the skin of a subject, said patch being administered to a subject Capable of releasing an effective amount of one or more triethylenetetramine active agents, the subject may be (1) diabetic or (II) copper levels capable of being reduced to fully or partially ameliorate or reverse one or more Systolic dysfunction, diastolic dysfunction, systolic abnormalities, recoil dysfunction, and ejection index disorder (which can be determined by ultrasonography, MRI, or other imaging techniques) and/or one or more of at least one of the causes of diabetic nephropathy, Diabetic nephropathy and/or damage caused by renal copper accumulation, and/or damage to at least the renal arteries, and/or selected from one or more of atrophy, loss of myocytes, expansion of the extracellular space, and extracellular matrix Structural damage to the heart with increased deposition of (and consequential) and/or structural damage to coronary arteries selected from damage (and consequences) to at least the medial (muscular) and intima (endothelial) layers of patients.

Another aspect of the invention includes an article of manufacture comprising a container containing a dosage form comprising one or more active agents as CR, SR and/or ER, or containing a pharmaceutically acceptable copper chelate Dosage forms of CR, SR and/or ER of the mixture, the copper chelating agent includes but not limited to one or more acceptable triethylenetetramine active agents; and for complete or partial improvement and/or reversal of test A description of a condition in a patient, wherein the subject is (I) a diabetic, or (II) a subject whose copper levels alleviate one or more of the above conditions.

Another aspect of the invention is an article of manufacture comprising a packaging material; a CR, SR and/or ER dosage form comprising one or more pharmaceutically acceptable triethylenetetramine active agents in the packaging material, wherein the packaging material Affixed with a label stating that the dosage form is useful for improving, reversing and/or restoring the health of a subject who is (I) diabetic, or (II) copper levels slow down any one or more of Subjects with the conditions described above.

In one embodiment of the dosage form, the effective amount and/or dosing regimen relates to an effective daily dose of the triethylenetetramine active agent (e.g. as triethylenetetramine dihydrochloride) capable of providing to the subject Salt calculation, regardless of whether the salt is contained in the dosage form unit) is 4g or less per day, even if it is taken orally, the dosage is 1mg-4g per day.

In another embodiment, the oral release dose (cumulative or otherwise) ranges from 200 mg to 4 g per day. In a further embodiment, the daily dose is 1.2 g or less.

In another aspect, the subject is provided with a dose (regardless of the amount in the dosage unit) of 1 mg per day, based on the salt of triethylenetetramine dihydrochloride or other substances described herein. -1.2g. If the oral administration dose is 200mg-1.2g per day.

In a further embodiment, the triethylenetetramine active agent is released in the dosage form unit at a pH of 7.2-7.6 (preferably a pH of 7.4±0.1).

In another embodiment of a dosage form such as triethylenetetramine active agent, such as triethylenetetramine dihydrochloride in a sustained release dosage form, the release of the active agent in the subject is always less than that used for Results of a 250 mg oral dosage form in Wilson's disease.

In another embodiment of a sustained release dosage form such as triethylenetetramine active agent, triethylenetetramine active agent such as triethylenetetramine dihydrochloride is suitable for once-daily administration and can be administered in vivo Slow or controlled and sustained release. Triethylenetetramine which releases no more than 10% of its dihydrochloride triethylenetetramine at an acidic pH of about < 4.5 within 5 hours and releases triethylenetetramine in a controlled manner on dissolution in vivo or in vitro The dihydrochloride exceeds 50% within 12 hours at a pH of approximately <6.5.

In another aspect of the present invention there is provided administration of effective Quantities of, for example, one or more triethylenetetramine active agents. In a specific embodiment, the DR, SR, ER, RA or CR preparation is suitable for the treatment of any of the aforementioned conditions, including but not limited to, heart failure, diabetic heart disease, acute coronary syndrome, hypertensive heart disease, Ischemic heart disease, coronary artery disease, peripheral artery disease, Wilson's disease, or any form of cancer. The DR, SR, ER, RA or CR formulation may comprise an effective dosage unit administered to a subject at a dose of about 1 mg to 600 mg per dose of at least one triethylenetetramine active agent, and in a further embodiment, The total daily dosage ranges from 5 g to 1 mg to maintain the required blood level of the triethylenetetramine active agent for a certain duration, preferably at least about 18-24 hours.

Another aspect of the present invention is, for example, a formulation of at least one triethylenetetramine active agent that maintains a constant plasma concentration of the triethylenetetramine active agent over a longer period of time and is effective from Copper removal in subjects with one or more of the conditions described herein, including, but not limited to, heart failure, diabetic heart disease, acute coronary syndrome, hypertensive heart disease, ischemia Heart disease, coronary artery disease, peripheral artery disease, Wilson's disease, or any form of cancer.

Another aspect of the invention includes a device comprising one or more triethylenetetramine active agents in an integral matrix device and useful for treating one or more of the symptoms described herein, including but not limited to, Heart failure, diabetic heart disease, acute coronary syndrome, hypertensive heart disease, ischemic heart disease, coronary artery disease, peripheral artery disease, Wilson's disease, or any form of cancer.

In a specific embodiment of the monolithic matrix device, the one or more triethylenetetramine active agents are contained in the dispersed soluble matrix, and the one or more triethylenetetramine active agents are The effect can be increased by dissolving or swelling in the matrix. Monolithic matrix devices may include, but are not limited to, one or more of the following excipients: hydroxypropyl cellulose (BP) or cellulose hydroxypropyl ether (USP); hydroxypropyl methylcellulose (BP, USP); cellulose (BP, USP); calcium carboxymethylcellulose (BP, USP); acrylic acid polymer or carboxypolyethylene (Carbopol) or carbomer (BP, USP); or linear glucuronate polymer Substances such as alginic acid (BP, USP), such as those formulated from the agglomeration system of alginic acid (and its salts)-gelatin colloids, or those lipids encapsulated by alginic acid and poly-L-lysine films body. In addition, the monolithic matrix includes one or more triethylenetetramine active agents dissolved in a soluble matrix, and the one or more triethylenetetramine active agents enter the matrix through microscopic channels in the aqueous solvent and Works after dissolving triethylenetetramine particles.

In a further specific embodiment, the monolithic matrix comprises, for example, one or more triethylenetetramine active agent particles as described, in a lipid matrix or an insoluble polymer matrix, including but not limited to carnauba wax (BP, USP); medium-chain triglycerides such as fractionated coconut oil (BP) or saturated triglyceride medium (PhEur); or preparations of cellulose ethyl ether or ethyl cellulose (BP, USP). The amount of lipid in said monolithic matrix may be 20-40% w/w of the hydrophobic solid matter. Lipids can remain intact during release.

In another specific embodiment, the device further comprises one or more of the following substances in addition to the one or more triethylenetetramine active agents such as described: a channeling agent, Such as sodium chloride or one or more sugars, which can leach out of the formulation, forming aqueous microporous channels (capillaries) through which solvents can enter and allow the drug to be released.

Alternatively, the device is any hydrophilic polymer matrix wherein said one or more active agents such as triethylenetetramine are concentrated in admixture with any water-swellable hydrophilic polymer.

For example the triethylenetetramine active agent is contained in the hydrophilic polymer matrix at a level of 20-80% (w/w).

In a specific embodiment, the hydrophilic polymer matrix comprises, in addition to one or more triethylenetetramine active agents, any one or more of the following: gel modifiers such as one or more Sugars, counterions, pH buffering agents, surfactants, lubricants such as magnesium stearate and/or glidants such as colloidal silicon dioxide.

Another aspect of the invention includes devices comprising an effective amount of one or more triethylenetetramine active agents such as described, comprising or comprising a rate-controlling membrane surrounding a drug depot, and comprising galactose and microcrystalline cellulose. The ratio of galactose to microcrystalline cellulose may be, for example, about 60:40.

The clinical trials described below demonstrate that a daily divided dose of 1.2 g of triethylenetetramine can be effectively maintained (involving transient drug levels in vivo) long-term for the purpose of ameliorating and/or reversing cardiac structural damage and/or coronary artery damage required high dose levels. Such a daily dose of 1.2 g can be provided by the application of 300 mg capsules of triethylenetetramine hydrochloride taken half an hour before meals, two capsules in the morning and two capsules in the evening.

Determination of free copper [equals total plasma copper minus ceruloplasmin-bound copper] is available on the Merck & Co Inc SYPRINERO (salt of triethylenetetramine dihydrochloride) capsule data sheet (www.Merck.com) to complete the procedure disclosed: which states that "The most reliable indicator for monitoring therapy is to determine free copper in serum at a value equal to the determined amount of total copper plus ceruloplasmin-bound copper." Poor. Adequately treated subjects typically have serum free copper less than 10mcg/dL. Treatment can be monitored by 24-hour urine analysis for copper. Urine must be collected in copper-free glassware. Due to low copper Diet can keep copper absorption below 1 mg per day, and a negative copper balance is ideal if the subject has 0.5-1.0 mg of copper in their urine collected over a 24-hour period."

Description of drawings

We rely on the study of triethylenetetramine dihydrochloride salts in the STZ rat model and humans, and hope to further describe the present invention by the following drawings:

Figure 1 shows the urinary excretion of diabetic and non-diabetic animals in response to increasing doses of TET or equal volumes of saline, wherein the urinary excretion of diabetic and non-diabetic animals responded to increasing doses of TET (bottom value; 0.1, 1.0, 10, 100 ^mg.kg in 75 μl of saline followed by 125 μl of saline flush, time-injected in the direction of the arrow) or an equal volume of saline (top value), and each point represents a 15-minute urine collection Period (see Example 2 for detailed methods); error indicates whether the SEM and P-value descriptions are significant (P<0.05).

Figure 2 shows the urinary excretion of non-diabetic and diabetic animals receiving increasing doses of TET or equal volumes of saline, where diabetic (top values) and non-diabetic rats (bottom values) received increasing doses of TET Urinary excretion of amines (0.1, 1.0, 10, 100 ^mg.kg in 75 μl saline followed by flushing with 125 μl saline, time-injected in the direction of the arrow) or an equal volume of saline, and each point represents a 15-minute Urine collection period (see Example 2 for detailed method); error indicated by SEM and P-value description is significant (P<0.05).

Figure 3 shows copper excretion in urine of diabetic and non-diabetic animals receiving increasing doses of triethylenetetramine or an equal volume of saline, wherein diabetic (top values) and non-diabetic rats (bottom values) received increasing doses of triethylenetetramine Copper excretion in urine with ethylenetetramine (0.1, 1.0, ¹⁰ , 100 mg.kg in 75 μl saline followed by flushing with 125 μl saline, timed injection in the direction of the arrow) or an equal volume of saline, and every Each point represents a 15-minute urine collection period (see Example 2 for detailed methods); error indicates whether the SEM and P-value describe significance (P<0.05).

Figure 4 shows the same information as in Figure 18 and illustrates the urinary copper excretion per gram of animal body weight in diabetic and non-diabetic animals in response to increasing doses of triethylenetetramine (bottom value : 0.1, 1.0, 10, 100 mg.kg ^-1 in 75 μl saline followed by 125 μl saline flush, timed injection in the direction of the arrow) or an equal volume of saline (top value), and each point represents 15 minutes of urine Collection period (see Example 2 for detailed methods); error indicates whether the SEM and P-value descriptions are significant (P<0.05).

Figure 5 shows the total copper excretion in non-diabetic and diabetic animals given saline or drugs, where the total urinary copper excretion (μmol) was given after saline (black bars, n=7) or triethylenetetramine ( Shaded bars, n=7) in non-diabetic animals, or in diabetic animals given saline (gray bars, n=7) or triethylenetetramine (white bars, n=7); error shown SEM and P-value description Whether it is significant (P<0.05).

Figure 6 shows total copper excretion per gram of body weight in animals receiving TET or saline (non-diabetic: shaded bars, n=7; diabetic: white bars, n=7) Total urinary copper excretion per gram of body weight (μmol.gBW ⁻¹ ) in animals with saline or saline (non-diabetic: black bars, n=7; diabetic: gray bars, n=7); errors are shown in SEM and P-values describe whether Significant (P<0.05).

Figure 7 shows urinary iron excretion in diabetic and non-diabetic animals receiving increasing doses of TET (0.1, 1.0 , 10, 100mg.kg ^-1 and then washed with 125μl of saline, injected according to the time in the direction of the arrow) or equal volume of saline diabetic rats (top value) and non-diabetic rats (bottom value) urinary iron excretion, and Each point represents a 15-minute urine collection period (see Example 2 for detailed methods); errors are indicated by SEM and P-values describe whether they are significant (P<0.05).

Figure 8 shows the urinary iron excretion per gram body weight of diabetic and non-diabetic animals receiving TET or saline, where the urinary copper excretion per gram body weight of diabetic and non-diabetic animals responded to increasing doses of TET (Bottom values: 0.1, 1.0, 10, 100 ^mg.kg in 75 μl saline followed by flushing with 125 μl of saline, time-injected in the direction of the arrow) or an equal volume of saline (top value), and each point represents 15 Minute urine collection period (see Example 2 for detailed method); error indicates whether the SEM and P value description is significant (P<0.05).

Figure 9 shows the total urinary iron excretion in non-diabetic and diabetic animals given saline or drug, where the total urinary iron excretion (μmol) was treated with saline (black bars, n=7) or triethylenetetramine ( Shaded bars, n=7) in non-diabetic animals, and in diabetic animals given saline (gray bars, n=7) or triethylenetetramine (white bars, n=7); error shown SEM and P-value description Whether it is significant (P<0.05).

Figure 10 shows total urinary ferric excretion per gram of body weight in animals receiving TET or saline (non-diabetic: shaded bars, n=7; diabetic: white bars, n=7 ) or saline (non-diabetic: black stripes, n=7; diabetic: gray stripes, n=7) total urinary iron excretion per gram of body weight (μmol.gBW ⁻¹ ); errors are shown in SEM and P-values are depicted Whether it is significant (P≤0.05).

Figure 11 shows urine [Cu] measured by AAS (△) and EPR (▲) after continuous administration of 10 mg.kg ^-1 (A) and 100 (B) triethylenetetramine pellets, as shown in Figure 19 ; (inset) Urine background corrected EPR signal from 75 min indicating the presence of Cu ^II -triethylenetetramine; ^*, P<0.05, ^** , P<0.01 vs. control.

Figure 12 is a table comparing copper and iron excretions of animals receiving triethylenetetramine or saline, and statistical analysis was performed using a mixed linear model.

Figure 13 shows the body weight of the animals in the experiment of Example 5 over time.

Figure 14 shows the glucose levels of the animals in the experiments of Example 5 over time.

Figure 15 is a graph showing animal cardiac output measured in Example 5.

Figure 16 is a graph showing the coronary blood flow of animals measured in Example 5.

Figure 17 is a graph showing coronary blood flow normalized to final heart weight as determined in Example 5.

Figure 18 is a graph showing animal aortic blood flow measured in Example 5.

Figure 19 is a graph showing the maximum positive change rate of the pressure of the ventricles of the animals for each cardiac cycle (systole) determined in Example 5.

Figure 20 is a graph showing the maximum rate of decrease in the pressure of the ventricles of the animals for each cardiac cycle (diastole) measured in Example 5.

Figure 21 shows the percentage of cardiac function retained after each load in animals determined in Example 5.

Figure 22 shows LV-myocardial structure of STZ-diabetic rats and matched non-diabetic control rats after 7 weeks of oral triethylenetetramine treatment, where heart sections were taken after functional studies. Each image represents 5 separate slices per heart, for a total of three hearts per treatment. ad is, 120-μM LV sections co-stained for actin (submuscarin-488, orange) and immunostained for β ₁ -integrin (CY5-paired secondary antibody, magenta) ( Scale stripes=33μm) laser focus image, a. untreated control group; b. untreated diabetic group; c. triethylenetetramine-treated diabetic group; d. triethylenetetramine-treated non-diabetic control group. eh. TEM images corresponding to 70-nM sections stained with uranyl acetate/lead citrate (scale bars = 158nm); e. untreated control group; f. untreated diabetic group; g. three B The diabetic group treated with ethylenetetramine; h. The non-diabetic control group treated with triethylenetetramine.

Figure 23 shows the effect of six-month oral administration of triethylenetetramine on LV mass in humans with T2DM, in which triethylenetetramine (600 mg twice daily) or corresponding placebo was given to diabetic patients (n=15) or corresponding Controls (n=15) were compared in a double-blind trial in which differences in LV mass (g; mean and 95% confidence interval) were determined using MRI of labeled hearts.

Figure 24 shows the effect of a randomized, double-blind, controlled trial comparing oral triethylenetetramine and placebo on urinary Cu excretion in men with uncomplicated T2DM and the corresponding non-diabetic control group, in which urinary Cu excretion (μmol.2h ^-1 Subjects with a single 2.4-g oral dose of triethylenetetramine and corresponding placebo on day 1 (as baseline) and day 7 are shown in Table 9, placebo-treated T2DM, ○, placebo-treated Control group, ●, T2DM treated with triethylenetetramine, □, control group treated with triethylenetetramine, ■. Cu excretion of T2DM treated with triethylenetetramine was significantly greater than that of T2DM treated with triethylenetetramine Non-diabetic control group (P<0.05).

Figure 25 shows the response of mean arterial pressure (MAP) in diabetic and non-diabetic animals flushed with 10 mg.kg ^- 1 triethylenetetramine in 75 μl + 125 μl saline (or an equivalent volume of saline). Each point represents the average value of the points collected every 2 seconds over a period of 1 minute. The timing of drug administration (or saline) is indicated in the direction of the arrow. error display SEM, and

Figure 26 shows the UV-Vis spectral scan of the TET formulation stored for 15 days with addition of copper to form the TET-copper complex. Scans were performed on day 0 (control formulation) and day 15. There were three formulations containing triethylenetetramine, one stored at 4°C in the dark, another at room temperature (21°C) in the dark, and a third at room temperature in daylight. Copper is added after the spectral scan is complete.

Detailed description of the invention

We describe the STZ rat model used to study human diabetic and non-diabetic subjects, and reduction of free copper can fully or partially ameliorate or reverse cardiac structural damage. This includes damage due to atrophy, loss of cardiomyocytes, expansion of the extracellular space and increased deposition of extracellular matrix (and consequences), and structural damage to the coronary arteries (and consequences). To demonstrate reversal of injury in STZ rats, as further described, relevant doses for humans were found involving urinary copper clearance. In addition, damage to cardiac structures under physiological conditions will sensitize distant stem cells, which can migrate to the injury site and undergo stem cell differentiation, ie, can promote structural and functional repair. However, accumulation of redox-active transition metals, especially copper, in cardiac tissue and coronary arteries of diabetic subjects has been shown to inhibit normal tissue regeneration through stem cell migration. In other words, elevated copper levels in the tissue suppressed the normal biological behavior of these undifferentiated cells. Even in non-diabetic mammals (e.g., without type 2 diabetes) and in mammals without abnormalities in glucose metabolism (e.g., without IGT or IFG), a reduction in extracellular copper favorably reduces and/or reverses copper-related damage. For example, overall or partial improvement of tissue repair by regeneration of normal tissue stem cell responses.

Evidence of principle in a 2-phase study showed positive results. However, when compared with its pharmacokinetic profile and recently discovered site-of-action profile, the dosing regimen is suboptimal. The bioavailability of active ingredients such as triethylenetetramine dihydrochloride following oral administration is low (<10%) due to poor absorption and first-pass metabolism. Both the dihydrochloride of triethylenetetramine and its metabolite, N-acetyl-triethyltetramine hydrochloride, are capable of binding copper, but the chelating activity of N-acetyl-triethyltetramine hydrochloride has been shown to be Reported to be significantly lower than triethylenetetramine dihydrochloride. See Kodama H., et al. Life Sciences 61:899-907 (1997). In addition, foods, mineral supplements, and other medications can adversely affect the absorption of triethylenetetramine dihydrochloride. The half-life of various copper chelating agents, such as triethylenetetramine, which can treat or reverse heart failure and coronary heart disease, is only a relatively short time of action of about 2 hours. Ideally, triethylenetetramine should be used at its maximum tolerated dose in the current treatment, and the dosage regimen should be suitable for its pharmacokinetics and the characteristics of its site of action. For plasma concentrations of triethylenetetramine administered orally, see Miyazaki, K., et al., "Determination of trientinein plasma of subjects with high-performance liquid chromatography," ChenzPharm Bull 38: 1035-38 (1998). Multiple dosing regimens are often used in subjects with heart failure and/or coronary artery disease. For the above-mentioned reasons, the dosage, dosage formulation and/or administration route of said dosage and preparation of copper chelating agents need to be improved.

The present invention relates to and provides novel dosages and formulations of copper chelating agents, such as triethylenetetramine active agents, and routes of administration of the various dosages and dosage forms. Triethylenetetramine active agents include triethylenetetramine, salts of triethylenetetramine, prodrugs of triethylenetetramine and salts of said prodrugs, triethylenetetramine analogs and said Salts and prodrugs of analogues, and/or metabolites of triethylenetetramine and salts and prodrugs of said metabolites, including but not limited to N-acetyl-triethylenetetramine, and N-acetyl Salts and prodrugs of triethylenetetramine. Not believed to be bound by theory, the dosage and formulation and route of administration provide unexpected beneficial effects in the overall or partial amelioration and reversal of the diseases, disorders and symptoms described herein, and copper is believed to play an important role therein.

Wilson's disease is an inherited defect in hepatobiliary copper excretion. Copper accumulation and copper toxicity lead to liver disease and, in some patients, brain damage. Most of the patients are between the ages of 10 and 40, presenting with liver disease, neurological disease of the ataxia type, or behavioral abnormalities, often concurrently. Wilson's disease can be effectively treated with oral copper chelators. Copper chelators have been shown to be excreted primarily in feces in Wilson's disease, with effective chelation occurring in the intestine (or inhibition of absorption), or by partial recovery mechanisms that allow excess copper to be excreted via urine or bile sweat, or both combination. See Siegemund R, et al., "Mode of action of triethylenetetramine dihydrochloride on copper metabolism in Wilson's disease," Acta Neurol Scand. 83(6):364-6 (June 1991).

In contrast, the experiments described here surprisingly showed that administration of the copper chelator triethylenetetramine dihydrochloride, eg, to non-Wilson's disease patients, did not result in increased excretion of copper in feces. See Example 9 and Table 11. Excess copper excretion in non-Wilson's disease subjects treated with copper chelators was primarily through urine rather than feces. See embodiment 8 and accompanying drawing 12. These data support the notion that systemically administered (parenteral) copper chelators are lower doses than oral doses, or that controlled release copper chelators are lower doses than oral doses, or that oral doses are more effective. The low dose avoids the undesired first-pass effect, allowing more of the active ingredient to act parenterally, with significant beneficial effects as described here. This includes administration of such doses and dosage forms that provide measurable release into the circulatory system (including intramuscular, intraperitoneal, subcutaneous and intravenous administration) compared to direct alimentary canal administration. ). Thus, the compounds may also be formulated for parenteral administration by injection (including, bolus injection or continuous infusion) and may be presented in preservative-added ampoules, prefilled syringes, small bolus containers or units in multidose containers. preparation.

According to the present invention, the dosage and dosage form of copper chelating agents such as triethylenetetramine, in order to maintain the desired blood concentration and tissue level, can be made into a preparation that efficiently removes copper from the body through urine, and a lower dosage than oral administration , which can more effectively treat the onset and progression of various pathologically increased or undesired copper-induced diseases. Such diseases include, but are not limited to, the following: heart failure, diabetic heart disease, acute coronary syndrome, hypertensive heart disease, ischemic heart disease, coronary heart disease, peripheral arterial disease, and can be treated with copper chelation agents of cancer.

Triethylenetetramine has a strongly basic part, containing multiple nitrogens, which can be converted into many appropriate acid addition salts with acid, for example, the calculated equivalent of triethylenetetramine and acid are reacted in an inert solvent , as in ethanol or water, and if the dosage form requires dry salts, distillation is performed. Usable acids are those which give physiologically acceptable salts. Nitrogen-containing copper chelating agents such as triethylenetetramine Active agents such as triethylenetetramine, may be released in the form of salts (such as acid addition salts, triethylenetetramine dihydrochloride) and act as copper chelating agents , can help the body to easily remove copper from the kidneys by forming stable soluble complexes. Inorganic acids such as sulfuric acid, nitric acid, hydrohalic acids such as hydrochloric acid or hydrobromic acid, phosphoric acids such as orthophosphoric acid, sulfamic acid can be used. The list is not exhaustive. Other organic acids can also be used, especially aliphatic, cycloaliphatic, araliphatic, aromatic or heterocyclic, mono- or polycarboxylic, sulfonic or sulfuric acids (e.g. formic acid, acetic acid, propane Acid, pivalic acid, diethylacetic acid, malonic acid, succinic acid, pimelic acid, fumaric acid, maleic acid, lactic acid, tartaric acid, malic acid, citric acid, gluconic acid, ascorbic acid, niacin, isonicotinic acid acid, methanesulfonic acid, ethanedisulfonic acid, 2-hydroxymethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, naphthalene mono- and disulphonic acid and lauryl sulfate). Suitable salt forms can be prepared by those skilled in the art. Nitrogen-containing copper chelating agents such as triethylenetetramine activators, such as triethylenetetramine, can also form quaternary ammonium salts, wherein the nitrogen atom is attached to a suitable organic group, such as alkyl, alkenyl, alkynyl or aralkyl. In a specific embodiment, the nitrogen-containing copper chelating agent is in the form of a solution and/or a suspension, which may contain a buffering agent whose pH value is close to neutral, which is much lower than the pH value of 14 of the triethylenetetramine solution.

Other triethylenetetramine active agents include derivatives of triethylenetetramine active agents, eg, triethylenetetramine in combination with ortho-picolinic acid (2-nicotinic acid). These derivatives include triethylpicoline and salts of triethylpicoline, such as triethylpicoline hydrochloride. Also included are triethyltetramine bis-picoline and salts of triethyltetramine bis-picoline, for example, triethyltetramine bis-picoline hydrochloride. The picolinic acid group can be linked to triethylenetetramine using conventional chemical techniques, such as one or more _CH2 group moieties. Those skilled in the art can prepare other suitable derivatives, for example, triethylenetetramine-PEG derivatives, which can be used in specific dosage forms for oral administration with enhanced bioavailability.

Other triethylenetetramine active agents include triethylenetetramine analog active agents. Such analogs include cyclic and acyclic analogs represented by the general formula:

式I

Formula I

The acyclic analogue of triethylenetetramine can be the following acyclic analogue based on the four-heteroatom described in the above general formula I, wherein X1, X2, X3 and X4 are independently selected from N, S or O atoms such that

(a) For the series of four nitrogens, that is, when X1, X2, X3 and X4 are N, then: R1, R2, R3, R4, R5 and R6 are independently selected from H, _CH3 , C2-C10 straight Chain or branched alkyl, C3-C10 cycloalkyl, C1-C6 alkylC3-C10 cycloalkyl, aryl, mono-, di-, tri-, tetra- and penta-substituted aryl, heteroaryl, fused Aryl, C1-C6 alkyl aryl, C1-C6 alkyl mono-, di-, tri-, tetra-, and penta-substituted aryl, C1-C5 alkyl heteroaryl, C1-C6 alkyl fused aryl, CH ₂ COOH, CH ₂ SO ₃ H, CH ₂ PO(OH) ₂ , CH ₂ P(CH ₃ )O(OH); n1, n2 and n3 are independently selected from 2 or 3; and, R7, R8, R9, R10, R11 and R12 are independently selected from H, CH ₃ , C2-C10 straight or branched chain alkyl, C3-C10 cycloalkyl, C1-C6 alkyl, C3-C10 cycloalkyl, aryl, mono, di , Tri-, tetra- and penta-substituted aryl, heteroaryl, fused aryl, C1-C6 alkyl aryl, C1-C6 alkyl mono-, di-, tri-, tetra- and penta-substituted aryl, C1-C5 Alkyl heteroaryl, C1-C6 alkyl fused aryl. Additionally, one or several of R1, R2, R3, R4, R5 or R6 may be functionalized to enable attachment to peptides, proteins, polyethylene glycol and other compounds capable of modifying the overall pharmacokinetics, drug release profile and/or half-life. chemical group. Examples of these functional groups include, but are not limited to, C1-C10 alkyl-CO-peptide, C1-C10 alkyl-CO-protein, C1-C10 alkyl-CO-PEG, C1-C10 alkyl-NH-peptide , C1-C10 Alkyl-NH-Protein, C1-C10 Alkyl-NH-CO-PEG, C1-C10 Alkyl-S-Peptide, C1-C10 Alkyl-S-Protein. Furthermore, one or several of R7, R8, R9, R10, R11 or R12 can be functionalized to allow attachment of peptides, proteins, polyethylene glycol and other chemical groups capable of modifying pharmacokinetics, drug release properties and/or half-life group. Examples of these functional groups include, but are not limited to, C1-C10 alkyl-CO-peptide, C1-C10 alkyl-CO-protein, C1-C10 alkyl-CO-PEG, C1-C10 alkyl-NH-peptide , C1-C10 alkyl-NH-protein, C1-C10 alkyl-NH-CO-PEG, C1-C10 alkyl-S-peptide, and C1-C10 alkyl-S-protein.

(b) For the first trinitrogen series, that is, when X1, X2, and X3 are N, and X4 is S or O, then: R6 does not exist; R1, R2, R3, R4, R5, and R6 are independent H, CH ₃ , C2-C10 linear or branched alkyl, C3-C10 cycloalkyl, C1-C6 alkyl, C3-C10 cycloalkyl, aryl, mono, di, tri, tetra and Penta-substituted aryl, heteroaryl, fused aryl, C1-C6 alkylaryl, C1-C6 alkyl mono-, di-, tri-, tetra-, and penta-substituted aryl, C1-C5 alkylheteroaryl , C1-C6 alkyl fused aryl, CH ₂ COOH, CH ₂ SO ₃ H, CH ₂ PO(OH) ₂ , CH ₂ P(CH ₃ )O(OH); n1, n2 and n3 are independently selected from 2 or 3; and, R7, R8, R9, R10, R11 and R12 are independently selected from H, CH ₃ , C2-C10 straight chain or branched chain alkyl, C3-C10 cycloalkyl, C1-C6 alkyl C3 -C10 cycloalkyl, aryl, mono-, di-, tri-, tetra-, and penta-substituted aryl, heteroaryl, fused aryl, C1-C6 alkylaryl, C1-C6 alkyl mono-, di-, tri- , Four and five substituted aryl, C1-C5 alkyl heteroaryl, C1-C6 alkyl fused aryl. Additionally, one or several of R1, R2, R3, R4, R5 can be functionalized to enable attachment to peptides, proteins, polyethylene glycol and other chemical groups capable of modifying pharmacokinetics, drug release properties and/or half-life. Examples of these functional groups include, but are not limited to, C1-C10 alkyl-CO-peptide, C1-C10 alkyl-CO-protein, C1-C10 alkyl-CO-PEG, C1-C10 alkyl-NH-peptide , C1-C10 Alkyl-NH-Protein, C1-C10 Alkyl-NH-CO-PEG, C1-C10 Alkyl-S-Peptide, C1-C10 Alkyl-S-Protein. Furthermore, one or several of R7, R8, R9, R10, R11 or R12 can be functionalized to allow attachment of peptides, protein PEGs and other chemical groups capable of modifying pharmacokinetics, drug release properties and/or half-life . Examples of these functional groups include, but are not limited to, C1-C10 alkyl-CO-peptide, C1-C10 alkyl-CO-protein, C1-C10 alkyl-CO-PEG, C1-C10 alkyl-NH-peptide , C1-C10 alkyl-NH-protein, C1-C10 alkyl-NH-CO-PEG, C1-C10 alkyl-S-peptide, and C1-C10 alkyl-S-protein.

(c) For the second trinitrogen series, that is, when X1, X2, and X4 are N, and X3 is O or S, then: R4 does not exist; R1, R2, R3, R5, and R6 are independently selected from H, CH ₃ , C2-C10 straight or branched chain alkyl, C3-C10 cycloalkyl, C1-C6 alkyl, C3-C10 cycloalkyl, aryl, mono-, di-, tri-, tetra- and penta-substituted Aryl, heteroaryl, fused aryl, C1-C6 alkylaryl, C1-C6 alkyl mono-, di-, tri-, tetra-, and penta-substituted aryl, C1-C5 alkylheteroaryl, C1 -C6 alkyl fused aryl, CH ₂ COOH, CH ₂ SO ₃ H, CH ₂ PO(OH) ₂ , CH ₂ P(CH ₃ )O(OH); n1, n2 and n3 are independently selected from 2 or 3; and, R7, R8, R9, R10, R11 and R12 are independently selected from H, CH ₃ , C2-C10 straight chain or branched chain alkyl, C3-C10 cycloalkyl, C1-C6 alkyl C3-C10 Cycloalkyl, aryl, mono-, di-, tri-, tetra-, and penta-substituted aryl, heteroaryl, fused aryl, C1-C6 alkylaryl, C1-C6 alkyl mono-, di-, tri-, tetra And five-substituted aryl, C1-C5 alkyl heteroaryl, C1-C6 alkyl fused aryl. Additionally, one or several of R1, R2, R3, R5 or R6 may be functionalized to enable attachment to peptides, proteins, polyethylene glycol and other chemical groups capable of modifying pharmacokinetics, drug release properties and/or half-life. Examples of these functional groups include, but are not limited to, C1-C10 alkyl-CO-peptide, C1-C10 alkyl-CO-protein, C1-C10 alkyl-CO-PEG, C1-C10 alkyl-NH-peptide , C1-C10 Alkyl-NH-Protein, C1-C10 Alkyl-NH-CO-PEG, C1-C10 Alkyl-S-Peptide, C1-C10 Alkyl-S-Protein. Furthermore, one or several of R7, R8, R9, R10, R11 or R12 can be functionalized to allow attachment of peptides, protein PEGs and other chemical groups capable of modifying pharmacokinetics, drug release properties and/or half-life . Examples of these functional groups include, but are not limited to, C1-C10 alkyl-CO-peptide, C1-C10 alkyl-CO-protein, C1-C10 alkyl-CO-PEG, C1-C10 alkyl-NH-peptide , C1-C10 alkyl-NH-protein, C1-C10 alkyl-NH-CO-PEG, C1-C10 alkyl-S-peptide, and C1-C10 alkyl-S-protein.

(d) For the series of the first dinitrogen, that is, when X2, and X3 are N, and X1 and X4 are O or S, then: R1, R6 do not exist; R2, R3, R4, and R5 are independently selected From H, CH ₃ , C2-C10 straight chain or branched alkyl, C3-C10 cycloalkyl, C1-C6 alkyl C3-C10 cycloalkyl, aryl, mono, di, tri, tetra and penta Substituted aryl, heteroaryl, fused aryl, C1-C6 alkylaryl, C1-C6 alkyl mono-, di-, tri-, tetra- and penta-substituted aryl, C1-C5 alkylheteroaryl, C1-C6 alkyl fused aryl, CH ₂ COOH, CH ₂ SO ₃ H, CH ₂ PO(OH) ₂ , CH ₂ P(CH ₃ )O(OH); n1, n2 and n3 are independently selected from 2 Or 3; and, R7, R8, R9, R10, R11 and R12 are independently selected from H, CH ₃ , C2-C10 straight chain or branched chain alkyl, C3-C10 cycloalkyl, C1-C6 alkyl C3- C10 cycloalkyl, aryl, mono-, di-, tri-, tetra- and penta-substituted aryl, heteroaryl, fused aryl, C1-C6 alkylaryl, C1-C6 alkyl mono-, di-, tri-, Four and five substituted aryl, C1-C5 alkyl heteroaryl, C1-C6 alkyl fused aryl. Additionally, one or several of R2, R3, R4 or R5 may be functionalized to enable attachment to peptides, proteins, polyethylene glycol and other chemical groups capable of modifying pharmacokinetics, drug release properties and/or half-life. Examples of these functional groups include, but are not limited to, C1-C10 alkyl-CO-peptide, C1-C10 alkyl-CO-protein, C1-C10 alkyl-CO-PEG, C1-C10 alkyl-NH-peptide , C1-C10 Alkyl-NH-Protein, C1-C10 Alkyl-NH-CO-PEG, C1-C10 Alkyl-S-Peptide, C1-C10 Alkyl-S-Protein. Furthermore, one or several of R7, R8, R9, R10, R11 or R12 can be functionalized to allow attachment of peptides, protein PEGs and other chemical groups capable of modifying pharmacokinetics, drug release properties and/or half-life . Examples of these functional groups include, but are not limited to, C1-C10 alkyl-CO-peptide, C1-C10 alkyl-CO-protein, C1-C10 alkyl-CO-PEG, C1-C10 alkyl-NH-peptide , C1-C10 alkyl-NH-protein, C1-C10 alkyl-NH-CO-PEG, C1-C10 alkyl-S-peptide, and C1-C10 alkyl-S-protein.

(e) For the second series of dinitrogen, that is, when X1, and X3 are N, and X2 and X4 are O or S, then: R3, R6 do not exist; R1, R2, R4, and R5 are independently selected From H, CH ₃ , C2-C10 straight chain or branched alkyl, C3-C10 cycloalkyl, C1-C6 alkyl C3-C10 cycloalkyl, aryl, mono, di, tri, tetra and penta Substituted aryl, heteroaryl, fused aryl, C1-C6 alkylaryl, C1-C6 alkyl mono-, di-, tri-, tetra- and penta-substituted aryl, C1-C5 alkylheteroaryl, C1-C6 alkyl fused aryl, CH ₂ COOH, CH ₂ SO ₃ H, CH ₂ PO(OH) ₂ , CH ₂ P(CH ₃ )O(OH); n1, n2 and n3 are independently selected from 2 Or 3; and, R7, R8, R9, R10, R11 and R12 are independently selected from H, CH ₃ , C2-C10 straight chain or branched chain alkyl, C3-C10 cycloalkyl, C1-C6 alkyl C3- C10 cycloalkyl, aryl, mono-, di-, tri-, tetra- and penta-substituted aryl, heteroaryl, fused aryl, C1-C6 alkylaryl, C1-C6 alkyl mono-, di-, tri-, Four and five substituted aryl, C1-C5 alkyl heteroaryl, C1-C6 alkyl fused aryl. Additionally, one or several of R1, R2, R4 or R5 may be functionalized to enable attachment to peptides, proteins, polyethylene glycol and other chemical groups capable of modifying pharmacokinetics, drug release properties and/or half-life. Examples of these functional groups include, but are not limited to, C1-C10 alkyl-CO-peptide, C1-C10 alkyl-CO-protein, C1-C10 alkyl-CO-PEG, C1-C10 alkyl-NH-peptide , C1-C10 alkyl-NH-protein, C1-C10 alkyl-NH-CO-PEG, C1-C10 alkyl-S-peptide, and C1-C10 alkyl-S-protein. Furthermore, one or several of R7, R8, R9, R10, R11 or R12 can be functionalized to allow attachment of peptides, protein PEGs and other chemical groups capable of modifying pharmacokinetics, drug release properties and/or half-life . Examples of these functional groups include, but are not limited to, C1-C10 alkyl-CO-peptide, C1-C10 alkyl-CO-protein, C1-C10 alkyl-CO-PEG, C1-C10 alkyl-NH-peptide , C1-C10 alkyl-NH-protein, C1-C10 alkyl-NH-CO-PEG, C1-C10 alkyl-S-peptide, and C1-C10 alkyl-S-protein.

(f) For the series of the third dinitrogen, that is, when X1 and X2 are N, and X3 and X4 are O or S, then: R4 and R6 do not exist; R1, R2, R3, and R5 are independently selected From H, CH ₃ , C2-C10 straight chain or branched alkyl, C3-C10 cycloalkyl, C1-C6 alkyl C3-C10 cycloalkyl, aryl, mono, di, tri, tetra and penta Substituted aryl, heteroaryl, fused aryl, C1-C6 alkylaryl, C1-C6 alkyl mono-, di-, tri-, tetra- and penta-substituted aryl, C1-C5 alkylheteroaryl, C1-C6 alkyl fused aryl, CH ₂ COOH, CH ₂ SO ₃ H, CH ₂ PO(OH) ₂ , CH ₂ P(CH ₃ )O(OH); n1, n2 and n3 are independently selected from 2 Or 3; and, R7, R8, R9, R10, R11 and R12 are independently selected from H, CH ₃ , C2-C10 straight chain or branched chain alkyl, C3-C10 cycloalkyl, C1-C6 alkyl C3- C10 cycloalkyl, aryl, mono-, di-, tri-, tetra- and penta-substituted aryl, heteroaryl, fused aryl, C1-C6 alkylaryl, C1-C6 alkyl mono-, di-, tri-, Four and five substituted aryl, C1-C5 alkyl heteroaryl, C1-C6 alkyl fused aryl. Additionally, one or several of R1, R2, R3 or R5 may be functionalized to enable attachment to peptides, proteins, polyethylene glycol and other chemical groups capable of modifying pharmacokinetics, drug release properties and/or half-life. Examples of these functional groups include, but are not limited to, C1-C10 alkyl-CO-peptide, C1-C10 alkyl-CO-protein, C1-C10 alkyl-CO-PEG, C1-C10 alkyl-NH-peptide , C1-C10 alkyl-NH-protein, C1-C10 alkyl-NH-CO-PEG, C1-C10 alkyl-S-peptide, and C1-C10 alkyl-S-protein. Furthermore, one or several of R7, R8, R9, R10, R11 or R12 can be functionalized to allow attachment of peptides, protein PEGs and other chemical groups capable of modifying pharmacokinetics, drug release properties and/or half-life . Examples of these functional groups include, but are not limited to, C1-C10 alkyl-CO-peptide, C1-C10 alkyl-CO-protein, C1-C10 alkyl-CO-PEG, C1-C10 alkyl-NH-peptide , C1-C10 alkyl-NH-protein, C1-C10 alkyl-NH-CO-PEG, C1-C10 alkyl-S-peptide, and C1-C10 alkyl-S-protein.

(g) For the fourth dinitrogen series, that is, when X1 and X4 are N, and X2 and X3 are O or S, then: R3 and R4 do not exist; R1, R2, R5 and R6 are independently selected from H, CH ₃ , C2-C10 straight or branched chain alkyl, C3-C10 cycloalkyl, C1-C6 alkyl, C3-C10 cycloalkyl, aryl, mono-, di-, tri-, tetra- and penta-substituted Aryl, heteroaryl, fused aryl, C1-C6 alkylaryl, C1-C6 alkyl mono-, di-, tri-, tetra-, and penta-substituted aryl, C1-C5 alkylheteroaryl, C1 -C6 alkyl fused aryl, CH ₂ COOH, CH ₂ SO ₃ H, CH ₂ PO(OH) ₂ , CH ₂ P(CH ₃ )O(OH); n1, n2 and n3 are independently selected from 2 or 3; and, R7, R8, R9, R10, R11 and R12 are independently selected from H, CH ₃ , C2-C10 straight chain or branched chain alkyl, C3-C10 cycloalkyl, C1-C6 alkyl C3-C10 Cycloalkyl, aryl, mono-, di-, tri-, tetra-, and penta-substituted aryl, heteroaryl, fused aryl, C1-C6 alkylaryl, C1-C6 alkyl mono-, di-, tri-, tetra And five-substituted aryl, C1-C5 alkyl heteroaryl, C1-C6 alkyl fused aryl. Additionally, one or several of R1, R2, R5, or R6 can be functionalized to enable attachment to peptides, proteins, polyethylene glycol, and other chemical groups capable of modifying pharmacokinetics, drug release properties, and/or half-life. Examples of such functional groups include, but are not limited to, C1-C10 alkyl-CO-peptide, C1-C10 alkyl-CO-protein, C1-C10 alkyl-CO-PEG, C1-C10 alkyl-NH-- Peptide, C1-C10 Alkyl-NH-Protein, C1-C10 Alkyl-NH-CO-PEG, C1-C10 Alkyl-S-Peptide, and C1-C10 Alkyl-S-Protein. Furthermore, one or several of R7, R8, R9, R10, R11 or R12 can be functionalized to allow attachment of peptides, protein PEGs and other chemical groups capable of modifying pharmacokinetics, drug release properties and/or half-life . Examples of these functional groups include, but are not limited to, C1-C10 alkyl-CO-peptide, C1-C10 alkyl-CO-protein, C1-C10 alkyl-CO-PEG, C1-C10 alkyl-NH-peptide , C1-C10 alkyl-NH-protein, C1-C10 alkyl-NH-CO-PEG, C1-C10 alkyl-S-peptide, and C1-C10 alkyl-S-protein.

Second, for the analogs of the four-heteroatom ring series, R1 and R6 are linked together by a bridging group to form (CR13R14)n4, and X1, X2, X3 and X4 are independently selected from N, S or, O makes

(a) For the series of four nitrogens, that is, when X1, X2, X3 and X4 are N, then: R2, R3, R4, and R5 are independently selected from H, CH ₃ , straight or branched chains of C2-C10 chain alkyl, C3-C10 cycloalkyl, C1-C6 alkyl C3-C10 cycloalkyl, aryl, mono-, di-, tri-, tetra- and penta-substituted aryl, heteroaryl, fused aryl, C1-C6 alkyl aryl, C1-C6 alkyl mono-, di-, tri-, tetra-, and penta-substituted aryl, C1-C5 alkyl heteroaryl, C1-C6 alkyl fused aryl, CH ₂ COOH, CH ₂ SO ₃ H, CH ₂ PO(OH) ₂ , CH ₂ P(CH ₃ )O(OH); n1, n2, n3 and n4 are independently selected from 2 or 3; and, R7, R8, R9, R10 , R11, R12, R13, and R14 are independently selected from H, CH ₃ , C2-C10 straight or branched chain alkyl, C3-C10 cycloalkyl, C1-C6 alkyl, C3-C10 cycloalkyl, aryl , mono-, di-, tri-, tetra-, and penta-substituted aryl, heteroaryl, fused aryl, C1-C6 alkyl aryl, C1-C6 alkyl mono-, di-, tri-, tetra-, and penta-substituted aryl , C1-C5 alkyl heteroaryl, C1-C6 alkyl fused aryl. Additionally, one or several of R2, R3, R4, or R5 can be functionalized to enable attachment to peptides, proteins, polyethylene glycol, and other chemical groups capable of modifying pharmacokinetics, drug release properties, and/or half-life. Examples of these functional groups include, but are not limited to, C1-C10 alkyl-CO-peptide, C1-C10 alkyl-CO-protein, C1-C10 alkyl-CO-PEG, C1-C10 alkyl-NH-peptide , C1-C10 Alkyl-NH-Protein, C1-C10 Alkyl-NH-CO-PEG, C1-C10 Alkyl-S-Peptide, C1-C10 Alkyl-S-Protein. Further, one or more of R7, R8, R9, R10, R11, R12, R13, or R14 can be functionalized to allow attachment of peptides, proteins, polyethylene glycol, and other compounds capable of modifying pharmacokinetics, drug release properties, and/or The half-life of a chemical group. Examples of these functional groups include, but are not limited to, C1-C10 alkyl-CO-peptide, C1-C10 alkyl-CO-protein, C1-C10 alkyl-CO-PEG, C1-C10 alkyl-NH-peptide , C1-C10 alkyl-NH-protein, C1-C10 alkyl-NH-CO-PEG, C1-C10 alkyl-S-peptide, and C1-C10 alkyl-S-protein.

(b) For the series of three nitrogens, that is, when X1, X2, X3 are N, and X4 is S or O, then: R5 does not exist; R2, R3, and R4 are independently selected from H, CH ₃ , C2- C10 straight or branched chain alkyl, C3-C10 cycloalkyl, C1-C6 alkyl, C3-C10 cycloalkyl, aryl, mono-, di-, tri-, tetra- and penta-substituted aryl, heteroaryl , Fused aryl, C1-C6 alkyl aryl, C1-C6 alkyl mono-, di-, tri-, tetra-, and penta-substituted aryl, C1-C5 alkyl heteroaryl, C1-C6 alkyl fused aryl group, CH ₂ COOH, CH ₂ SO ₃ H, CH ₂ PO(OH) ₂ , CH ₂ P(CH ₃ )O(OH); n1, n2, n3 and n4 are independently selected from 2 or 3; and, R7 , R8, R9, R10, R11, R12, R13 and R14 are independently selected from H, CH ₃ , C2-C10 straight or branched chain alkyl, C3-C10 cycloalkyl, C1-C6 alkyl C3-C10 ring Alkyl, aryl, mono-, di-, tri-, tetra-, and penta-substituted aryl, heteroaryl, fused aryl, C1-C6 alkylaryl, C1-C6 alkyl mono-, di-, tri-, tetra-, and Penta-substituted aryl, C1-C5 alkyl heteroaryl, C1-C6 alkyl fused aryl. Additionally, one or several of R2, R3, or R4 can be functionalized to enable attachment to peptides, proteins, polyethylene glycol, and other chemical groups capable of modifying pharmacokinetics, drug release properties, and/or half-life. Examples of these functional groups include, but are not limited to, C1-C10 alkyl-CO-peptide, C1-C10 alkyl-CO-protein, C1-C10 alkyl-CO-PEG, C1-C10 alkyl-NH-peptide , C1-C10 alkyl-NH-protein, C1-C10 alkyl-NH-CO-PEG, C1-C10 alkyl-S-peptide, and C1-C10 alkyl-S-protein. Further, one or more of R7, R8, R9, R10, R11, R12, R13, or R14 can be functionalized to allow attachment of peptides, proteins, polyethylene glycol, and other compounds capable of modifying pharmacokinetics, drug release properties, and/or The half-life of a chemical group. Examples of these functional groups include, but are not limited to, C1-C10 alkyl-CO-peptide, C1-C10 alkyl-CO-protein, C1-C10 alkyl-CO-PEG, C1-C10 alkyl-NH-peptide , C1-C10 alkyl-NH-protein, C1-C10 alkyl-NH-CO-PEG, C1-C10 alkyl-S-peptide, and C1-C10 alkyl-S-protein.

(c) For the first dinitrogen series, that is, when X2, and X3 are N, and X1 and X4 are O or S, then: R2 and R5 do not exist; R3 and R4 are independently selected from H, CH ₃ , C2-C10 straight chain or branched chain alkyl, C3-C10 cycloalkyl, C1-C6 alkyl C3-C10 cycloalkyl, aryl, mono-, di-, tri-, tetra- and penta-substituted aryl groups, Heteroaryl, fused aryl, C1-C6 alkyl aryl, C1-C6 alkyl mono-, di-, tri-, tetra-, and penta-substituted aryl, C1-C5 alkyl heteroaryl, C1-C6 alkyl Fused aryl, CH ₂ COOH, CH ₂ SO ₃ H, CH ₂ PO(OH) ₂ , CH ₂ P(CH ₃ )O(OH); n1, n2, n3 and n4 are independently selected from 2 or 3; And, R7, R8, R9, R10, R11, R12, R13 and R14 are independently selected from H, CH ₃ , C2-C10 straight chain or branched chain alkyl, C3-C10 cycloalkyl, C1-C6 alkyl C3 -C10 cycloalkyl, aryl, mono-, di-, tri-, tetra-, and penta-substituted aryl, heteroaryl, fused aryl, C1-C6 alkylaryl, C1-C6 alkyl mono-, di-, tri- , Four and five substituted aryl, C1-C5 alkyl heteroaryl, C1-C6 alkyl fused aryl. Additionally, one or both of R3, and R4 can be functionalized to enable attachment to peptides, proteins, polyethylene glycol, and other chemical groups capable of modifying pharmacokinetics, drug release properties, and/or half-life. Examples of these functional groups include, but are not limited to, C1-C10 alkyl-CO-peptide, C1-C10 alkyl-CO-protein, C1-C10 alkyl-CO-PEG, C1-C10 alkyl-NH-peptide , C1-C10 alkyl-NH-protein, C1-C10 alkyl-NH-CO-PEG, C1-C10 alkyl-S-peptide, and C1-C10 alkyl-S-protein. Further, one or several of R7, R8, R9, R10, R11, R12, R13 or R14 can be functionalized to allow attachment of peptides, proteins PEG and other compounds capable of modifying pharmacokinetics, drug release properties and/or half-life chemical groups. Examples of these functional groups include, but are not limited to, C1-C10 alkyl-CO-peptide, C1-C10 alkyl-CO-protein, C1-C10 alkyl-CO-PEG, C1-C10 alkyl-NH-peptide , C1-C10 alkyl-NH-protein, C1-C10 alkyl-NH-CO-PEG, C1-C10 alkyl-S-peptide, and C1-C10 alkyl-S-protein.

(d) For the second dinitrogen series, that is, when X1, and X3 are N, and X2 and X4 are O or S, then: R3 and R5 do not exist; and R2 and R4 are independently selected from H, CH ₃ , C2-C10 straight chain or branched chain alkyl, C3-C10 cycloalkyl, C1-C6 alkyl C3-C10 cycloalkyl, aryl, mono-, di-, tri-, tetra- and penta-substituted aryl groups , heteroaryl, fused aryl, C1-C6 alkyl aryl, C1-C6 alkyl mono-, di-, tri-, tetra-, and penta-substituted aryl, C1-C5 alkyl heteroaryl, C1-C6 alkane fused aryl group, CH ₂ COOH, CH ₂ SO ₃ H, CH ₂ PO(OH) ₂ , CH ₂ P(CH ₃ )O(OH); n1, n2, n3 and n4 are independently selected from 2 or 3 and, R7, R8, R9, R10, R11, R12, R13 and R14 are independently selected from H, CH ₃ , C2-C10 straight or branched chain alkyl, C3-C10 cycloalkyl, C1-C6 alkyl C3-C10 cycloalkyl, aryl, mono-, di-, tri-, tetra- and penta-substituted aryl, heteroaryl, fused aryl, C1-C6 alkylaryl, C1-C6 alkyl mono-, di-, Three, four and five substituted aryl, C1-C5 alkyl heteroaryl, C1-C6 alkyl fused aryl. Additionally, one or both R2, or R4 can be functionalized to enable attachment to peptides, proteins, polyethylene glycol, and other chemical groups capable of modifying pharmacokinetics, drug release properties, and/or half-life. Examples of these functional groups include, but are not limited to, C1-C10 alkyl-CO-peptide, C1-C10 alkyl-CO-protein, C1-C10 alkyl-CO-PEG, C1-C10 alkyl-NH-peptide , C1-C10 alkyl-NH-protein, C1-C10 alkyl-NH-CO-PEG, C1-C10 alkyl-S-peptide, and C1-C10 alkyl-S-protein. Further, one or several of R7, R8, R9, R10, R11, R12, R13 or R14 can be functionalized to allow attachment of peptides, proteins PEG and other compounds capable of modifying pharmacokinetics, drug release properties and/or half-life chemical groups. Examples of these functional groups include, but are not limited to, C1-C10 alkyl-CO-peptide, C1-C10 alkyl-CO-protein, C1-C10 alkyl-CO-PEG, C1-C10 alkyl-NH-peptide , C1-C10 alkyl-NH-protein, C1-C10 alkyl-NH-CO-PEG, C1-C10 alkyl-S-peptide, and C1-C10 alkyl-S-protein.

(e) For a nitrogen series, that is, when X1 is N, and X2, X3 and X4 are O or S, then: R3, R4 and R5 do not exist; R2 is independently selected from H, CH ₃ , C2-C10 straight-chain or branched-chain alkyl, C3-C10 cycloalkyl, C1-C6 alkyl, C3-C10 cycloalkyl, aryl, mono-, di-, tri-, tetra-, and penta-substituted aryl, heteroaryl, Fused aryl, C1-C6 alkyl aryl, C1-C6 alkyl mono-, di-, tri-, tetra-, and penta-substituted aryl, C1-C5 alkyl heteroaryl, C1-C6 alkyl fused aryl , CH ₂ COOH, CH ₂ SO ₃ H, CH ₂ PO(OH) ₂ , CH ₂ P(CH ₃ )O(OH); n1, n2, n3 and n4 are independently selected from 2 or 3; and R7, R8 , R9, R10, R11, R12, R13 and R14 are independently selected from H, CH ₃ , C2-C10 straight chain or branched chain alkyl, C3-C10 cycloalkyl, C1-C6 alkyl C3-C10 cycloalkyl , aryl, mono-, di-, tri-, tetra-, and penta-substituted aryl, heteroaryl, fused aryl, C1-C6 alkylaryl, C1-C6 alkyl mono-, di-, tri-, tetra-, and penta-substituted Aryl, C1-C5 alkyl heteroaryl, C1-C6 alkyl fused aryl. Additionally, R2 can be functionalized to enable attachment to peptides, proteins, polyethylene glycol, and other chemical groups capable of modifying pharmacokinetics, drug release properties, and/or half-life. Examples of these functional groups include, but are not limited to, C1-C10 alkyl-CO-peptide, C1-C10 alkyl-CO-protein, C1-C10 alkyl-CO-PEG, C1-C10 alkyl-NH-peptide , C1-C10 alkyl-NH-protein, C1-C10 alkyl-NH-CO-PEG, C1-C10 alkyl-S-peptide, and C1-C10 alkyl-S-protein. Further, one or several of R7, R8, R9, R10, R11, R12, R13 or R14 can be functionalized to allow attachment of peptides, proteins PEG and other compounds capable of modifying pharmacokinetics, drug release properties and/or half-life chemical groups. Examples of these functional groups include, but are not limited to, C1-C10 alkyl-CO-peptide, C1-C10 alkyl-CO-protein, C1-C10 alkyl-CO-PEG, C1-C10 alkyl-NH-peptide , C1-C10 alkyl-NH-protein, C1-C10 alkyl-NH-CO-PEG, C1-C10 alkyl-S-peptide, and C1-C10 alkyl-S-protein.

Formula II

Acyclic analogues of three-heteroatoms such as general formula II, wherein X1, X2, and X3 are independently selected from N, S or O atoms, such that

(a) For the series of three nitrogens, that is, when X1, X2, and X3 are N, then: R1, R2, R3, R5 and R6 are independently selected from H, CH ₃ , C2-C10 straight chain or branched chain alkyl, C3-C10 cycloalkyl, C1-C6 alkyl C3-C10 cycloalkyl, aryl, mono-, di-, tri-, tetra- and penta-substituted aryl, heteroaryl, fused aryl, C1-C6 alkyl aryl, C1-C6 alkyl mono-, di-, tri-, tetra-, and penta-substituted aryl, C1-C5 alkyl heteroaryl, C1-C6 alkyl fused aryl, CH ₂ COOH, CH ₂ SO ₃ H, CH ₂ PO(OH) ₂ , CH ₂ P(CH ₃ )O(OH); n1 and n2 are independently selected from 2 or 3; and, R7, R8, R9, and R10 are independently selected from From H, CH ₃ , C2-C10 straight chain or branched chain alkyl, C3-C10 cycloalkyl, C1-C6 alkyl C3-C10 cycloalkyl, aryl, mono-, di-, tri-, tetra- and penta-substituted Aryl, heteroaryl, fused aryl, C1-C6 alkyl aryl, C1-C6 alkyl mono-, di-, tri-, tetra-, and penta-substituted aryl, C1-C5 alkyl heteroaryl, C1- C6 alkyl fused aryl. Additionally, one or several of R1, R2, R3, R5 or R6 may be functionalized to enable attachment to peptides, proteins, polyethylene glycol and other chemical groups capable of modifying pharmacokinetics, drug release properties and/or half-life. Examples of these functional groups include, but are not limited to, C1-C10 alkyl-CO-peptide, C1-C10 alkyl-CO-protein, C1-C10 alkyl-CO-PEG, C1-C10 alkyl-NH-peptide , C1-C10 alkyl-NH-protein, C1-C10 alkyl-NH-CO-PEG, C1-C10 alkyl-S-peptide, and C1-C10 alkyl-S-protein. Furthermore, one or more of R7, R8, R9, or R10 can be functionalized to allow attachment of peptides, protein polyethylene glycols, and other chemical groups capable of modifying pharmacokinetics, drug release properties, and/or half-life. Examples of these functional groups include, but are not limited to, C1-C10 alkyl-CO-peptide, C1-C10 alkyl-CO-protein, C1-C10 alkyl-CO-PEG, C1-C10 alkyl-NH-peptide , C1-C10 alkyl-NH-protein, C1-C10 alkyl-NH-CO-PEG, C1-C10 alkyl-S-peptide, and C1-C10 alkyl-S-protein.

(b) For the first dinitrogen series, that is, when X1 and X3 are N, and X2 is S or O, then: R3 does not exist; R1, R2, R3, R5, and R6 are independently selected from H , CH ₃ , C2-C10 straight chain or branched chain alkyl, C3-C10 cycloalkyl, C1-C6 alkyl C3-C10 cycloalkyl, aryl, mono-, di-, tri-, tetra- and penta-substituted Aryl, heteroaryl, fused aryl, C1-C6 alkylaryl, C1-C6 alkyl mono-, di-, tri-, tetra-, and penta-substituted aryl, C1-C5 alkylheteroaryl, C1- C6 alkyl fused aryl, CH ₂ COOH, CH ₂ SO ₃ H, CH ₂ PO(OH) ₂ , CH ₂ P(CH ₃ )O(OH); n1 and n2 are independently selected from 2 or 3; and , R7, R8, R9, and R10 are independently selected from H, CH ₃ , C2-C10 straight or branched chain alkyl, C3-C10 cycloalkyl, C1-C6 alkyl, C3-C10 cycloalkyl, aryl , mono-, di-, tri-, tetra-, and penta-substituted aryl, heteroaryl, fused aryl, C1-C6 alkyl aryl, C1-C6 alkyl mono-, di-, tri-, tetra-, and penta-substituted aryl , C1-C5 alkyl heteroaryl, C1-C6 alkyl fused aryl. Additionally, one or several of R1, R2, R5, R6 can be functionalized to enable attachment to peptides, proteins, polyethylene glycol and other chemical groups capable of modifying pharmacokinetics, drug release properties and/or half-life. Examples of these functional groups include, but are not limited to, C1-C10 alkyl-CO-peptide, C1-C10 alkyl-CO-protein, C1-C10 alkyl-CO-PEG, C1-C10 alkyl-NH-peptide , C1-C10 alkyl-NH-protein, C1-C10 alkyl-NH-CO-PEG, C1-C10 alkyl-S-peptide, and C1-C10 alkyl-S-protein. Furthermore, one or several of R7, R8, R9 or R10 can be functionalized to allow attachment of peptides, protein polyethylene glycols and other chemical groups capable of modifying pharmacokinetics, drug release properties and/or half-life. Examples of these functional groups include, but are not limited to, C1-C10 alkyl-CO-peptide, C1-C10 alkyl-CO-protein, C1-C10 alkyl-CO-PEG, C1-C10 alkyl-NH-peptide , C1-C10 alkyl-NH-protein, C1-C10 alkyl-NH-CO-PEG, C1-C10 alkyl-S-peptide, and C1-C10 alkyl-S-protein.

(c) For the second dinitrogen series, that is, when X1 and X2 are N, and X3 is O or S, then: R3 does not exist; R1, R2, R5, and R6 are independently selected from H, CH ₃ , C2-C10 straight chain or branched chain alkyl, C3-C10 cycloalkyl, C1-C6 alkyl C3-C10 cycloalkyl, aryl, mono-, di-, tri-, tetra- and penta-substituted aryl groups , heteroaryl, fused aryl, C1-C6 alkyl aryl, C1-C6 alkyl mono-, di-, tri-, tetra-, and penta-substituted aryl, C1-C5 alkyl heteroaryl, C1-C6 alkane fused aryl group, CH ₂ COOH, CH ₂ SO ₃ H, CH ₂ PO(OH) ₂ , CH ₂ P(CH ₃ )O(OH); n1 and n2 are independently selected from 2 or 3; and, R7 , R8, R9 and R10 are independently selected from H, CH ₃ , C2-C10 straight or branched chain alkyl, C3-C10 cycloalkyl, C1-C6 alkyl, C3-C10 cycloalkyl, aryl, single, Di-, tri-, tetra- and penta-substituted aryl, heteroaryl, fused aryl, C1-C6 alkyl aryl, C1-C6 alkyl mono-, di-, tri-, tetra- and penta-substituted aryl, C1- C5 alkyl heteroaryl, C1-C6 alkyl fused aryl. Additionally, one or several of R1, R2, R5, or R6 can be functionalized to enable attachment to peptides, proteins, polyethylene glycol, and other chemical groups capable of modifying pharmacokinetics, drug release properties, and/or half-life. Examples of these functional groups include, but are not limited to, C1-C10 alkyl-CO-peptide, C1-C10 alkyl-CO-protein, C1-C10 alkyl-CO-PEG, C1-C10 alkyl-NH-peptide , C1-C10 alkyl-NH-protein, C1-C10 alkyl-NH-CO-PEG, C1-C10 alkyl-S-peptide, and C1-C10 alkyl-S-protein. Furthermore, one or more of R7, R8, R9, or R10 can be functionalized to allow attachment of peptides, protein polyethylene glycols, and other chemical groups capable of modifying pharmacokinetics, drug release properties, and/or half-life. Examples of these functional groups include, but are not limited to, C1-C10 alkyl-CO-peptide, C1-C10 alkyl-CO-protein, C1-C10 alkyl-CO-PEG, C1-C10 alkyl-NH-peptide , C1-C10 alkyl-NH-protein, C1-C10 alkyl-NH-CO-PEG, C1-C10 alkyl-S-peptide, and C1-C10 alkyl-S-protein.

The second is a three-heteroatom acyclic analogue of general formula II, wherein R1 and R6 are connected together by a bridging group to form (CR11R12)n3, and X1, X2, and X3 are independently selected from N , S or O atoms, such that

(a) For the series of three nitrogens, that is, when X1, X2, and X3 are N, then: R2, R3, and R5 are independently selected from H, CH ₃ , C2-C10 linear or branched chain alkane radical, C3-C10 cycloalkyl, C1-C6 alkyl C3-C10 cycloalkyl, aryl, mono-, di-, tri-, tetra-, and penta-substituted aryl, heteroaryl, fused aryl, C1-C6 Alkylaryl, C1-C6 alkyl mono-, di-, tri-, tetra-, and penta-substituted aryl, C1-C5 alkyl heteroaryl, C1-C6 alkyl fused aryl, CH ₂ COOH, CH ₂ SO ₃ H, CH ₂ PO(OH) ₂ , CH ₂ P(CH ₃ )O(OH); n1, n2 and n3 are independently selected from 2 or 3; and, R7, R8, R9, R10, R11, and R12 independently selected from H, CH ₃ , C2-C10 straight or branched chain alkyl, C3-C10 cycloalkyl, C1-C6 alkyl, C3-C10 cycloalkyl, aryl, mono, di, tri, tetra and Penta-substituted aryl, heteroaryl, fused aryl, C1-C6 alkylaryl, C1-C6 alkyl mono-, di-, tri-, tetra-, and penta-substituted aryl, C1-C5 alkylheteroaryl , C1-C6 alkyl fused aryl. Additionally, one or several of R2, R3, or R5 can be functionalized to enable attachment to peptides, proteins, polyethylene glycol, and other chemical groups capable of modifying pharmacokinetics, drug release properties, and/or half-life. Examples of these functional groups include, but are not limited to, C1-C10 alkyl-CO-peptide, C1-C10 alkyl-CO-protein, C1-C10 alkyl-CO-PEG, C1-C10 alkyl-NH-peptide , C1-C10 alkyl-NH-protein, C1-C10 alkyl-NH-CO-PEG, C1-C10 alkyl-S-peptide, and C1-C10 alkyl-S-protein. Furthermore, one or more of R7, R8, R9, R10, R11, or R12 can be functionalized to allow attachment of peptides, protein polyethylene glycols, and other chemical groups capable of modifying pharmacokinetics, drug release properties, and/or half-life group. Examples of these functional groups include, but are not limited to, C1-C10 alkyl-CO-peptide, C1-C10 alkyl-CO-protein, C1-C10 alkyl-CO-PEG, C1-C10 alkyl-NH-peptide , C1-C10 alkyl-NH-protein, C1-C10 alkyl-NH-CO-PEG, C1-C10 alkyl-S-peptide, and C1-C10 alkyl-S-protein.

(b) For the series of dinitrogen, that is, when X1, X2 is N, and X3 is S or O, then: R5 does not exist; R2 and R3 are independently selected from H, CH ₃ , straight chains of C2-C10 or Branched Alkyl, C3-C10 Cycloalkyl, C1-C6 Alkyl, C3-C10 Cycloalkyl, Aryl, Mono-, Di-, Tri-, Tetra-, and Penta-substituted Aryl, Heteroaryl, Fused Aryl , C1-C6 alkyl aryl, C1-C6 alkyl mono-, di-, tri-, tetra-, and penta-substituted aryl, C1-C5 alkyl heteroaryl, C1-C6 alkyl fused aryl, CH ₂ COOH , CH ₂ SO ₃ H, CH ₂ PO(OH) ₂ , CH ₂ P(CH ₃ )O(OH); n1, n2 and n3 are independently selected from 2 or 3; and, R7, R8, R9, R10, R11 and R12 are independently selected from H, CH ₃ , C2-C10 straight or branched chain alkyl, C3-C10 cycloalkyl, C1-C6 alkyl, C3-C10 cycloalkyl, aryl, mono, di, tri , four and five substituted aryl, heteroaryl, fused aryl, C1-C6 alkylaryl, C1-C6 alkyl mono-, di-, tri-, tetra- and penta-substituted aryl, C1-C5 alkyl Heteroaryl, C1-C6 alkyl fused aryl. Additionally, one or both R2 or R3 can be functionalized to enable attachment to peptides, proteins, polyethylene glycol and other chemical groups capable of modifying pharmacokinetics, drug release properties and/or half-life. Examples of these functional groups include, but are not limited to, C1-C10 alkyl-CO-peptide, C1-C10 alkyl-CO-protein, C1-C10 alkyl-CO-PEG, C1-C10 alkyl-NH-peptide , C1-C10 alkyl-NH-protein, C1-C10 alkyl-NH-CO-PEG, C1-C10 alkyl-S-peptide, and C1-C10 alkyl-S-protein. Furthermore, one or more of R7, R8, R9, R10, R11, or R12 can be functionalized to allow attachment of peptides, protein polyethylene glycols, and other chemical groups capable of modifying pharmacokinetics, drug release properties, and/or half-life group. Examples of these functional groups include, but are not limited to, C1-C10 alkyl-CO-peptide, C1-C10 alkyl-CO-protein, C1-C10 alkyl-CO-PEG, C1-C10 alkyl-NH-peptide , C1-C10 alkyl-NH-protein, C1-C10 alkyl-NH-CO-PEG, C1-C10 alkyl-S-peptide, and C1-C10 alkyl-S-protein.

(c) For a nitrogen series, that is, when X1 is N, and X2 and X3 are O or S, then: R3 and R5 do not exist; R2 is selected from H, CH ₃ , straight or branched chains of C2-C10 respectively chain alkyl, C3-C10 cycloalkyl, C1-C6 alkyl C3-C10 cycloalkyl, aryl, mono-, di-, tri-, tetra- and penta-substituted aryl, heteroaryl, fused aryl, C1-C6 alkyl aryl, C1-C6 alkyl mono-, di-, tri-, tetra-, and penta-substituted aryl, C1-C5 alkyl heteroaryl, C1-C6 alkyl fused aryl, CH ₂ COOH, CH ₂ SO ₃ H, CH ₂ PO(OH) ₂ , CH ₂ P(CH ₃ )O(OH); n1, n2 and n3 are independently selected from 2 or 3; and, R7, R8, R9, R10, R11 , and R12 are independently selected from H, CH ₃ , C2-C10 straight chain or branched chain alkyl, C3-C10 cycloalkyl, C1-C6 alkyl C3-C10 cycloalkyl, aryl, mono, di, tri , four and five substituted aryl, heteroaryl, fused aryl, C1-C6 alkylaryl, C1-C6 alkyl mono-, di-, tri-, tetra- and penta-substituted aryl, C1-C5 alkyl Heteroaryl, C1-C6 alkyl fused aryl. Additionally, R2 can be functionalized to enable attachment to peptides, proteins, polyethylene glycol, and other chemical groups capable of modifying pharmacokinetics, drug release properties, and/or half-life. Examples of these functional groups include, but are not limited to, C1-C10 alkyl-CO-peptide, C1-C10 alkyl-CO-protein, C1-C10 alkyl-CO-PEG, C1-C10 alkyl-NH-peptide , C1-C10 alkyl-NH-protein, C1-C10 alkyl-NH-CO-PEG, C1-C10 alkyl-S-peptide, and C1-C10 alkyl-S-protein. Furthermore, one or more of R7, R8, R9, R10, R11, or R12 can be functionalized to allow attachment of peptides, protein polyethylene glycols, and other chemical groups capable of modifying pharmacokinetics, drug release properties, and/or half-life group. Examples of these functional groups include, but are not limited to, C1-C10 alkyl-CO-peptide, C1-C10 alkyl-CO-protein, C1-C10 alkyl-CO-PEG, C1-C10 alkyl-NH-peptide , C1-C10 alkyl-NH-protein, C1-C10 alkyl-NH-CO-PEG, C1-C10 alkyl-S-peptide, and C1-C10 alkyl-S-protein.

The analogs of the present invention can be synthesized, isolated and purified by any method known in the art.

The present invention includes controlled release or other drug dosage and dosage form delivery formulations and devices comprising one or more copper chelating agents such as triethylenetetramine or salts thereof. The present invention includes, for example, doses and dosage forms, at least oral administration, transdermal administration, topical application, suppository release, mucosal release, injection (including subcutaneous administration, intramuscular administration, implant administration and intravenous administration ( bolus injection, slow intravenous injection and intravenous drip)), infusion devices (including implantable infusion devices, both active and passive), inhalation or insufflation administration, buccal administration, sublingual administration and Administration through the eye.

The diseases that described dose, dosage form and route of administration are applied include diabetic cardiomyopathy, diabetic acute coronary syndrome (such as myocardial infarction MI), diabetic hypertensive cardiomyopathy, acute Coronary artery syndrome, acute coronary syndrome associated with fasting glucose impairment (IFG), hypertensive cardiomyopathy associated with IGT, hypertensive cardiomyopathy associated with IFG, ischemic cardiomyopathy associated with IGT , ischemic cardiomyopathy associated with IFG, ischemic cardiomyopathy associated with coronary heart disease coronary atherosclerosis (CHD), myocardial disorders (cardiomyopathy or myocarditis) including idiopathic cardiomyopathy, metabolic cardiomyopathy (including diabetes cardiomyopathy, alcoholic cardiomyopathy, drug-induced cardiomyopathy, ischemic cardiomyopathy, and hypertensive cardiomyopathy), acute coronary syndrome unrelated to abnormal glucose metabolism, hypertensive cardiomyopathy unrelated to abnormal glucose metabolism Cardiomyopathy, ischemic cardiomyopathy unrelated to abnormal glucose metabolism (regardless of whether ischemic cardiomyopathy is related to coronary artery disease or coronary heart disease) and disease of one or more vascular trees, including aorta, carotid Arteries and disorders including arterial cerebral arteries, coronary arteries, renal arteries, retinal arteries, iliac arteries, femoral arteries, popliteal arteries, vasa vasa, arteriolar trees, and capillary beds, of major vessels (macrovascular disease) Atherosclerotic disease such as disorders of the aorta, coronary, carotid, cerebral, renal, iliac, femoral, and popliteal arteries, including but not limited to atrophy, loss of myocytes, extracellular space expansion, and extracellular Damage to (and consequences of) cardiac structures with increased deposition of matrix and/or coronary artery structures selected from damage (and consequences) of at least the middle layer (muscular layer) and/or the intimal layer (endothelial layer) Injuries, plaque rupture of atherosclerotic lesions of major vessels such as aortic, coronary, carotid, cerebral, renal, iliac, femoral, and popliteal arteries, systolic dysfunction, diastolic dysfunction , systolic disturbances, recoil characteristics and ejection index, toxicity, drug-induced and metabolic abnormalities including hypertension and/or diabetic disorders of small vessels (microvascular disease) such as retinal arterioles, glomerular arterioles, vasa vasa , cardiac arterioles and associated diseases of the eye, kidneys, heart and capillary beds of the central and peripheral nervous systems. Accordingly, the present invention also provides dosages and formulations of one or more novel copper chelating agents such as triethylenetetramine or salts thereof for use in the treatment of the diseases described herein in humans or other mammals. The application of these doses, dosage forms and devices can effectively treat the above diseases, and can be used for administration to humans and other mammals.

The present invention provides drug release dosages comprising one or more copper chelating agents such as triethylenetetramine or salts thereof. Accordingly, the present invention is directed, in part, to novel delivery doses containing one or more copper chelators such as triethylenetetramine for optimal bioavailability and maintenance of plasma levels in the therapeutic range, including prolonged duration of action. , and lead to an increase in the maintenance time of the blood drug concentration of one or more copper chelating agents such as triethylenetetramine or its salts, ensuring an ideal therapeutic dose range at the site of action. Controlled-release formulations also provide optimal drug concentration at the site of action and reduce treatment cycles.

The invention also provides drug delivery formulations and devices comprising one or more copper chelating agents such as one or more triethylenetetramine active agents, including but not limited to triethylenetetramine, triethylenetetramine Dihydrochloride or other pharmaceutically acceptable salts, the formulation is suitable for periodic administration, including once-a-day administration, providing a controlled and/or low-dose long-acting copper chelate release in vivo. complexing agent, and the excretion of copper chelating agent is excreted through urine.

The present invention also provides drug release formulations and devices comprising one or more copper chelating agents such as one or more triethylenetetramine active agents, including but not limited to triethylenetetramine, triethylenetetramine The dihydrochloride or other pharmaceutically acceptable salts, suitable for periodic dosing, including once-a-day administration, provide increased bioavailability of the copper chelating agent, and excretion of the copper chelating agent is urinary excretion.

Examples of controlled release dosages for releasing the drugs of the present invention can be found in Sweetman, S.C. (Ed.). Martindale. The Complete Drug Reference, 33rd Edition, Pharmaceutical Press, Chicago, 2002, 2483 pp.; Aulton, M.E. (Ed. .) Pharmaceutics. The Science of Dosage Form Design. Churchill Livingstone, Edinburgh, 2000, 734 pp.; and, Ansel, H.C., Allen, L.V. and Popovich, N.G. Pharmaceutical Dosage Forms and Drug Delivery Systems, 7th Ed., Lippincott 1999, 676 pp. Excipients for the preparation of drug delivery systems are described in numerous publications and are routine techniques for those skilled in the art, see Kibbe, E.H. Handbook of Pharmaceutical Excipients, 3rd Ed., American Pharmaceutical Association, Washington, 2000, 665 pp .. The USP also provides examples of oral sustained-release dosage forms, including those excipients that make tablets and capsules. See, for example, USP 23/National Drug Collection 18, United States Pharmacopeia Commission, Inc., Rockville MD, 1995 (referred to herein as USP), also describing a specific test for determining the amount released from extended-release and delayed-release tablets and capsules test. The USP test for determining extended release and delayed release is based on the dissolution of drug from the dosage unit over the test time. Various testing instruments and operating procedures can be found in USP. Certain monographs include compliance with specific criteria for the assays, devices, and procedures used. The examples give, for example, the release of aspirin in aspirin extended release tablets (see e.g. Ansel, H.C., Allen, L.V. and Popovich, N.G. Pharmaceutical Dosage Forms and Drug Delivery Systems, 7th Ed., Lippincott 1999, p.237). Sustained-release tablets and capsules shall be in accordance with the standards of uniformity described in USP customary dosage units. The uniformity of dosage units can be determined by two methods, the weight variance method or the fill volume variance method, as described in the USP. Further analyzes involving extended release dosage forms are provided by the F.D.A (see Guidance for Industry. Extended release oral dosage forms: development, evaluation, and application of in vitro/in vivo correlations. Rockville, MD: Center for Drug Evaluation and Research, US Food and Drug Administration Drug Administration, 1997). Adherence to the dosing regimen is always necessary in order to be able to arrive at an optimal treatment regimen. When compared with BID (twice a day), TID (three times a day), QID (four times a day), etc., the multi-dose oral administration regimen of triethylenetetramine doses used so far for the treatment of Wilson's disease, the present invention recognizes Another advantage of a dosage unit that provides the above release levels is the lower dosage of triethylenetetramine.

The present invention also includes various drug delivery systems for delivering one or more copper chelating agents such as triethylenetetramine or salts thereof. Accordingly, the present invention describes a novel drug delivery system. These include, Modified Release dosage forms (MR) of the present invention, including Delayed Release dosage forms (DR); Sustained Release dosage forms (PA); Controlled Release dosage forms (CR); Extended Release dosage forms (ER); Timed Release dosage forms (TR); And long-acting release dosage form (LA). Mostly, these terms are used to describe oral dosage forms, but the term rate-controlled delivery can be applied to certain types of drug delivery systems where delivery is controlled by characteristics of the device rather than by physiological or environmental conditions, such as gastrointestinal tract pH , or the time it takes the drug to move through the gastrointestinal tract. These formulations affect the following aspects: (1) delayed total drug release for a certain period of time after administration, (2) drug release at short intervals after administration, (3) drug release in a controlled manner under the dosing of the delivery system. Slow rate release, (4) drug release at a constant rate, and/or (5) significantly longer drug release time than conventional formulations. Where "moderate", "delayed", "slow", "extended", "timed", "long-acting", "controlled" and/or "extended" release dosage forms herein may be any suitable release dosage form.

Advantages of dosing formulations of one or more copper chelators such as triethylenetetramine or salts thereof include convenience to the subject; increased compliance; and stability achieved with up to twice daily dosing. constant drug levels over prolonged periods of time; prophylaxis of drug toxicity; and clearance of treatment failure, especially at night. Sustained release dosage forms of the present invention include dosage forms in which drug release characteristics are based on time, course and/or site of administration, which cannot be achieved by conventional or rapid release dosage forms. See, e.g., Bogner, R.H. Bioavailability and bioequivalence of extended-release oral dosage forms. U.S Pharmacist 22 (Suppl.): 3-12 (1997); Scale-up of oral extended-release drug delivery systems: part 1, overview. Pharmaceutical Manufacturing 2: 23-27 (1985). Extended-release dosage forms of the present invention include, for example, those described in the US Food and Drug Administration (F.D.A.), which allow for less frequent dosing than conventional dosage forms, such as solutions, or immediate-release dosage forms. See Bogner, R.H. Bioavailability and bioequivalence of extended-release oral dosage forms. US Pharmacist 22 (Suppl.): 3-12 (1997); Guidance for industry. Oral dosage forms: development, evaluation, and application of the invitro/in vivo correlations. Rockville, MD: Center for Drug Evaluation and Research, US Food and Drug Administration (1997). Repeated active dosage forms of the invention include, for example, dosage forms containing two single doses of drug, one for immediate release and the other for sustained release. In extended release dosage forms, bilayer tablets may be prepared, with one layer for immediate release and the other for sustained release. Targeted release dosage forms of the present invention include, for example, those that facilitate drug release and targeted concentration of drug activity in an area, tissue or location of the body for specific absorption or ease of use.

One example is oral delivery dosage forms, i.e. tablets, capsules, lozenges, etc., or liquid dosage forms such as syrups, aqueous solutions, emulsions, etc., which provide sustained release of active ingredients such as one or more of the present Invented compounds and formulations.

Examples of transdermal dosage units for the delivery of compounds and formulations of the invention include transdermal patches, skin bandages, and the like.

Examples of topical dosage units for the delivery of compounds and formulations of the invention include lotions, sticks, sprays, ointments, pastes, creams, gels and the like, administered directly through the skin or through a vehicle, They provide a slow release of the active ingredient during delivery to the subject through vehicles including pads, patches, and the like.

Examples of suppository dosage units for the delivery of compounds and formulations of the invention include any solid dosage form capable of being inserted into a body orifice, particularly rectal, vaginal and urethral dosage forms.

Examples of transmucosal dosage units for delivering the compounds and formulations of the invention include depositing solutions for enemas, pessaries, tampons, creams, gels, pastes, foams, spray solutions, powders, etc., containing Suitable carriers for the active substance are known to those skilled in the art.

Examples of injectable dosage units for delivery of the compounds and formulations of the invention include single or multiple bolus injections, which may be administered intravenously, subcutaneously, and intramuscularly, or may be administered orally.

Examples of depot dosage units for delivery of compounds and formulations of the invention include active ingredient-containing pellets or cartridges or solid dosage forms wherein the active ingredient is disposed in a biodegradable polymer matrix, microemulsion matrix, lipid in the body matrix, or encapsulated in the form of microcapsules.

Examples of infusion devices for delivering the compounds and formulations of the invention include infusion pumps containing one or more copper chelating agents such as triethylenetetramine or salts thereof, which can be administered in a desired dose or steady state, Also included are implantable drug pumps. Implantable infusion devices of the present invention include any solid form in which the active substance is encapsulated or dispersed in a biodegradable polymer, or a synthetic polymer, silicone, silicone rubber, silicone rubber or the like polymer.

Examples of dosage units for inhalation or insufflation to deliver the compounds and formulations of the invention include compositions comprising solutions and/or suspensions and/or powders in pharmaceutically acceptable aqueous or organic solvents or mixtures thereof.

Examples of buccal administration dosage units for delivering the compounds of the present invention and formulations include lozenges, tablets, etc., solutions and/or suspensions and/or powders in pharmaceutically acceptable aqueous or organic solvents or mixtures thereof combination.

Examples of sublingual dosage units for delivery of compounds and formulations of the present invention include lozenges, tablets, etc., solutions and/or suspensions and/or powders in pharmaceutically acceptable aqueous or organic solvents or mixtures thereof. combination.

Examples of ophthalmic dosage units for delivery of the compounds and formulations of the present invention include solutions and/or suspensions in pharmaceutically acceptable aqueous or organic solvents or mixtures thereof, or compositions of implants.

The present invention provides dosage delivery devices, and formulations comprising one or more copper chelating agents such as triethylenetetramine or salts thereof, with one or more suitable anions to form of complexes. An example of a sustained release dosage form comprising one or more copper chelating agents such as triethylenetetramine or its salts is by incorporating the active ingredients in certain complexes such as those with the anions of various forms of tannic acid Formed complexes (see, for example, Merck Index 12th Ed., 9221). The dissolution of these complexes is dependent on the pH of the environment. The slow dissolution rate provides extended release of the drug. For example, the tannate salt of triethylenetetramine provides this property and can be used to treat disease conditions associated with increased copper. Examples of equivalent products are referred to those trade names compound phenylephrine dose Rynatan (Wallace: see Madan, P.L., "Sustained release dosage forms," U.S. Pharmacist 15:39-50 (1990); Ryna-12 S, which contains a mixture of mepyramine tannate with phenylephrine tannate, Martindale 33rd Ed., 2080.4).

The present invention also includes coated beads, granules or microspheres comprising one or more copper chelating agents such as triethylenetetramine or salts thereof. Accordingly, the present invention also provides a method of obtaining sustained release properties of one or more copper chelating agents such as triethylenetetramine or salts thereof by placing the drug in coated beads, granules or microspheres . Such formulations of one or more copper chelating agents such as triethylenetetramine or salts thereof may be used in the treatment of corresponding diseases in humans or other mammals. In such systems, the drug is distributed on beads, pellets, granules or other specialized systems. Using conventional pan coating or air suspension coating techniques, the drug substrate solution is placed on top of inert seeds or beads formed from sugar and starch or microcrystalline cellulose pellets. Seed crystals typically range in diameter from 425-850 microns, while microcrystalline fiber pellets range from 170-600 microns (see Ansel, H.C., Allen, L.V. and Popovich, N.G. Pharmaceutical Dosage Forms and Drug Delivery Systems, 7th Ed., Lippincott 1999 , p.232). Microcrystalline cellulose spheres are considered to be more durable than sugar cores (see Celphere microcrystalline cellulosespheres. Philadelphia: FMC Corporation, 1996). Methods for the preparation of suitable drug-releasing microspheres have been published (see Arshady, R. Microspheres and microcapsules: a survey of manufacturing techniques. 1: suspension and cross-linking. Polymer Eng Sci 30: 1746-1758 (1989); see also Arshady , R. Microspheres and microcapsules: a survey of manufacturing techniques. 2: coacervation. Polymer Eng Sci 30: 905-914 (1990); see also Arshady R. Microspheres and micro-capsules: a survey of manufacturing techniques. 3 : solvent evaporation. Polymer Eng Sci 30: 915-924 (1990)). In the case of bolus drugs, the material of the starting particles may be formed by the drug itself. A portion of these granules can remain uncoated to provide rapid release of the drug. Other granules (approximately two-thirds to three-quarters) are coated with a liquid material such as beeswax, carnauba wax, glyceryl monostearate, cetyl alcohol, or a cellulosic material such as ethylcellulose (see below ). Next, the coated granules of different thicknesses are mixed to obtain a mixture with the desired drug release characteristics. The coating material may be dyed with one or more dyes to distinguish particles or beads of different thickness (by shade of color) and to provide product differentiation. After suitable mixing, the granules can be placed in capsules or compressed into tablets. Various coating systems are commercially available, including those based on water and those using ethylcellulose and plasticizers as coating materials (e.g., AquacoatTM [FMC Corporation, Philadelphia] and SurereleaseTM [Colorcon]; AquacoatTM [Colorcon]; aqueous polymeric dispersion. Philadelphia: FMC Corporation, 1991; Surerelease aqueous controlled release coating system. West Point, PA: Colorcon, 1990; Butler, J., Cumming, I, Brown, J. et al. A novel multiunit controlled-release system. Pharm Tech 22: 122-138 (1998); Yazici, E., Oner, L., Kas, H.S. & Hincal, A.A. Phenytoin sodium microspheres: bench scale formulation, process characterization and release kinetics. Pharmaceut Dev Technol 1: 175-183 (1996) ). Aqueous-based coating systems eliminate the hazards and environments associated with organic solvent-based systems. Comparable water and organic solvent coating methods (see Hogan, J.E. Aqueous versus organic solvent coating. Int J Pharm Tech Prod Manufacture 3: 17-20 (1982)). The thickness of the coating and the type of coating material can affect the rate at which the drug dissolves and penetrates the coating in body fluids. In general, the thicker the coating, the greater the resistance to penetration and the more delayed drug release and dissolution. Typically 1 mm diameter coated beads. Typically there are three or four release groups in combination, wherein more than 100 beads are loaded into the dosage unit (see Madan, P.L. Sustained release dosage forms. U.S. Pharmacist 15:39-50 (1990)). This provides coated beads with varying sustained or extended release rates and desired segmental orientation in the gastrointestinal tract. An example of such a dosage form is Spansule™ (SmithKline Beecham Corporation, U.K.). Examples of film-forming polymers that can be used in the water-insoluble slow release intermediate layer (for pellets, pellets or tablet cores) include ethyl cellulose, polyvinyl acetate, Eudragit® RS, Eudragit® RL, etc. (per Eudragit® RS and Eudragit(R) RL are ammonium methacrylate copolymers). The release rate can be controlled by combining with suitable water-soluble pore-forming substances such as lactose, mannitol, sorbitol, etc., or by the thickness of the coating layer. Multiplex tablets include pellets compressed in the form of pellets, 3-4 mm in diameter, and may be placed in gelatin capsules to give a suitable release profile. Each capsule may contain 8-10 minitablets, some uncoated for immediate release and others coated for extended drug release.

The following method can be used to produce a delivery system comprising one or more sustained release dosage forms of a copper chelator such as triethylenetetramine or a salt thereof or other triethylenetetramine active agent, suitable for use in humans and other Oral administration in mammals. Two basic mechanisms are required here to achieve sustained release of drug delivery. These alter the solubility and dispersibility of drugs and excipients. Herein, for example, four procedures may be applied, either simultaneously or sequentially. As follows: (i) hydration of the device (e.g., swelling of the matrix); (ii) diffusion of water into the device; (iii) controlled or delayed dissolution of the drug; and (iv) controlled or delayed diffusion of the dissolved drug out. Continuous release is zero-order kinetics and results from a constant rate of diffusion or osmosis. Sustained-release dosage forms are generally suitable for one of three types of systems: monolithic or single matrix; depot or membrane-controlled; or osmotic pump systems. Each comprises the following ingredients: active drug; release controlling agent; matrix modifier; drug modifier; supplemental coating agent; ed.) Handbook of Pharmaceutical Excipients, 3rd Edition, American Pharmaceutical Association, 2000, 665 pp.).

For oral dosage forms of the compounds and formulations of the invention, prolonged drug action can be achieved by affecting the rate at which the drug is released from the dosage form and/or by slowing the time it takes for the drug to pass through the gastrointestinal tract (see Bogner, R.H. Bioavailability and bioequivalence of extended-release oral dosage forms. US Pharmacist 22 (Suppl.): 3-12 (1997)). The rate of drug release from a solid dosage form can be modulated by processes such as: 1) regulating the solubility of the drug by controlling the action of biological fluids on the drug by applying a coating barrier; 2) controlling the diffusion of the drug from the dosage form; and 3) The chemical reaction or influence between a drug or its pharmaceutical barrier and a site-specific biological fluid. Such a system is also described here. In one aspect, the coating or encapsulation of the active ingredient can slow down digestion or dispersion in the intestinal tract with digestion as the release mechanism. The rate of availability of the active agent is a function of the rate of digestion of the dispersible material. Thus, the rate of release, and thus the effect of the agent, will vary depending on the subject's ability to digest the substance. In another aspect, as disclosed in U.S. Patent No. 3,247,066, the active agent is dispersed in a water-soluble colloid and then coated with a breakable plastic, non-digestible material that is permeable by diffusion of water. After ingestion and food passage through the gastrointestinal tract, water from body fluids disperses through the coating and enters causing swelling of the colloid. The coating ruptures due to swelling of the colloid and the entire content of the active agent is released. Although the rate of release varied little between subjects, initial plasma concentrations of total active agent were high and declined rapidly over time.

U.S. Patent No. 3,115,441 discloses another method of encapsulation for the delivery of the compounds and formulations of the present invention, wherein the particles of the active agent are initially thinly coated with a film-forming material and a non-toxic hydrophobic material, Coating is then carried out continuously with a material resistant to organic solvents. The coated granules are admixed with the uncoated active agent and the admixture is then formed into tablets by placing the coated tablet in a matrix of the uncoated active agent. Tablets made in this way have the advantage of rapid release of the compounds and formulations of the invention due to the rapid dissolution of the matrix material (comprising the initial dose) after ingestion.

Another aspect, as in U.S. Patent No. 4,025,613, is to provide an improved blood level of the compounds and formulations of the present invention due to the simple application of a thin film of a non-aqueous solution of cellulose acetate to the active agent. Granules (before compression) and untreated active agent granules form tablets that are dried to form a cellulose acetate coating. According to the film coating, those skilled in the art can select the appropriate film-forming agent: selected derivatives of cellulose such as hydroxypropyl methylcellulose (HPMC), ethylcellulose, acetylcellulose phthalate, levulinic acid Polymers and copolymers of cellulose, trimelliate, methacrylic acid and their derivatives. The film-forming agent can be added together with the following components: plasticizers (such as high molecular weight polyethylene glycol, polybasic acid esters such as citric acid or phthalic acid), fillers (such as talcum powder, metal oxides such as titanium dioxide ), selected from those pigments that can be used in the pharmaceutical and food industries.

Another slow release dosage form of the compounds and formulations of the invention is any suitable osmotic system having a semipermeable membrane of cellulose acetate, cellulose acetate butyrate, cellulose acetate propionate to control the release of the active agent. These can be dispersed coated with an aqueous enteric film without altering the release rate. An example of such an osmotic system is an osmotic pump device, which may be the Oros ^™ device from Alza Corporation (USA). The system consists of a die surrounded by a semipermeable membrane with 0.4mm diameter holes formed by a laser beam. The tablet core has two layers, one containing the drug (the active layer) and the other containing the polymeric osmotic agent (the push layer). The tablet core consists of active drug, fillers, viscosity modifiers and solubilizers. The system operates on the principle of osmotic pressure. The system is suitable for the release of a wide range of drugs, including triethylenetetramine or its salts. The coating process is simple and the release is zero order kinetics. When a tablet is ingested, the semipermeable membrane allows aqueous fluid from the stomach to enter the core, dissolving or suspending the drug. As pressure builds up to the osmotic layer, it presses or pumps the drug solution through the orifice to the edge of the tablet. Only a solution of the drug (not insoluble drugs) is able to pass through the orifice of the tablet. The system only requires a few drops of water per hour into the tablet. The rate of aqueous fluid influx and tablet functionality is dependent on the existence of an osmotic gradient between the contents of the bilayer tablet and the fluid in the gastrointestinal tract. Drug release is essentially continuous under a constant osmotic gradient. The rate of drug release can be varied by varying the surface area, thickness of the membrane of the composition and/or diameter of the orifice. The rate of drug release was not affected by gastrointestinal acidity, alkalinity, feeding conditions, or gastrointestinal motility. The biologically inactive components of the tablet remain intact during intestinal transit and are excreted in the feces as insoluble waste. Other examples of applications of this technology can be found in Glucotrol XL Extended Release Tablets (Pfizer Inc.) and Procardia XL Extended Release Tablets (Pfizer Inc.; see, Martindale 33rd Ed., p. 2051.3).

The present invention also provides release devices for the release of the compounds and formulations of the present invention employing an integral matrix comprising, for example, a slowly erodible or hydrophilic polymer matrix incorporating one or more compounds such as triethylenetetramine or a salt thereof. The copper chelating agent is pressed or embedded in the matrix.

Integral matrix devices for the release of the compounds and formulations of the invention include those formed using systems such as: (I) drug particles dispersed in a soluble matrix where the action is enhanced upon dissolution or swelling of the matrix; examples include hydrocolloids Bases such as hydroxypropylcellulose (BP) or hydroxypropylcellulose (USP); hydroxypropylmethylcellulose (HPMC; BP, USP); methylcellulose (MC; BP, USP); carboxymethylcellulose Calcium cellulose (calcium CMC; BP, USP); acrylic acid polymers or carboxypolymethylene (Carbopol) or carbomer (BP, USP); or linear glucuronic acid polymers such as seaweed Acids (BP, USP), such as those formed from microparticles of the alginic acid (alginate)-gelatin hydrocolloid agglomeration system, or those liposomes coated with a film of alginic acid and poly-L-lysine. The drug is released upon swelling of the polymer, which forms a matrix layer that can control the diffusion of the aqueous liquid to the core and thus the rate of diffusion of the drug in the system. In such systems, the rate of drug release depends on the natural tortuosity of the channels in the gel, as well as the viscosity of the liquid, and different release kinetics can be obtained, such as zero-order kinetics, or first-order kinetics combined with pulsed release . If the gel is not cross-linked, there is a weaker, non-fixed link between the polymer chains, which is dependent on secondary bonding. With such devices, high loadings of active drug can be achieved, as well as frequent efficient mixing. The device contains 20-80% drug (w/w), as well as gel modifiers that enhance drug diffusion; examples of such modifiers include sugars that increase the rate of hydration, ions that affect cross-linking , and pH buffers that can affect the level of polymer ionization. Hydrophilic matrix devices typically contain pH buffers, surfactants, counter ions, lubricants such as magnesium stearate (BP, USP) and glidants such as colloidal bismuth in addition to the drug and the hydrophilic matrix. Silica (USP; Anhydrous colloidal silica, BP); (II), drug particles dissolved in an insoluble matrix, where the drug enters the matrix through the solvent, usually through pores, and is obtained by dissolving the drug particles. Examples include systems formed from lipid matrices or insoluble polymer matrices, including formulations formed from carnauba wax (BP; USP); medium-chain triglycerides such as fractionated coconut oil (BP) or triglyceride-saturated media (PhEur) ; or cellulose ether or ethyl cellulose (BP, USP). Lipid matrices are easy and simple to prepare and can be mixed with the following powdered components: lipids (20-40% hydrophobic solids w/w) to remain intact during release; drugs; channeling agents such as chlorinated Sodium or sugar, can leach from the formulation, forming aqueous microporous channels (capillaries) through which the drug is released. In another system, using an insoluble polymer matrix, the drug is embedded in an inert insoluble polymer and released by leaching with an aqueous liquid that diffuses through the channels into the particle core so that the drug can be released from the device. The release rate is controlled by the degree of compression, particle size, natural properties and corresponding excipients (w/w). An example of such a device is Ferrous Gradumet (Martindale 33rd Ed., 1360.3). Further suitable examples of insoluble matrices are inert plastic matrices. By this method, the triethylenetetramine active agent is granulated with a plastic material, such as polyethylene, polyvinyl acetate, or polymethacrylate, and mixtures thereof, and then compressed into tablets. Once ingested, the drug is slowly released from the inert plastic matrix by diffusion (see Bodmeier, R. & Paeratakul, O., "Drug release from laminated polymeric films prepared from aqueous latexes," J PharmSci 79:32-26 (1990); Laghoueg, N., et al. "Oral polymer-drug devices with a core and an erodable shell for constant drug delivery," Int J Pharm50: 133-139 (1989); Buckton, G., et al. release from acrylic matrices." Int J Pharm 74:153-158 (1991)). Compression of tablets creates a matrix or plastic form that can retain its shape as the drug leaches and passes through the gastrointestinal tract. The immediate release portion of the drug can be compressed into the tablet surface. The inert tablet matrix is the waste product of the drug, which is excreted in feces. An example of a dosage form of this type is Gradumet (Abbott; see Ferro-Gradumet, Martindale 33rd Ed., p. 1860.4).

A further application is to combine the compounds and formulations of the present invention with polymer matrices in a pendent appendix (see Scholsky, K.M. & Fitch, R.M. Controlled release of pendant bioactive materials from acrylic polymercolloids. J Controlled Release 3: 87-108 (1986)). In these devices, the drug is incorporated via esters attached to polyacrylate latex particles, prepared by aqueous emulsion polymerization.

Further embodiments incorporate dosage forms of the compounds and formulations of the present invention wherein the drug is bonded to a biocompatible polymer via a labile chemical bond such as from a substituted anhydride (which itself is via an acid chloride to the drug: methacryloyl chloride and methoxy The polyanhydride bond prepared by the reaction of sodium benzoate) was used to form the matrix of the secondary polymer (Eudragit RL), which was released by hydrolysis in intestinal fluid (see Chaffi, N., Monthheard, J.P. & Vergnaud, J.M. Release of 2-aminothiazole from polymeric carriers. Int J Pharm 67: 265-274 (1992)).

In forming a suitable hydrophilic matrix for use in the compounds and formulations of the invention, the selected polymer must be capable of forming a gelatinous layer rapidly enough to protect the tablet core from too rapid breakdown by digestion. While increasing the proportion of polymer in the formulation, the increase of gel viscosity will lead to a decrease in the rate of drug diffusion and release (see Formulating for controlled release with Methocel Premium cellulose ethers. Midland, MI: Dow Chemical Company, 1995). Typically, 20% (w/w) of HPMC results in a satisfactory release rate of the drug from an extended release tablet formulation. However, as with all formulations, the possible effects of other formulation ingredients, such as fillers, tablet binders and disintegrants, must be taken into account. An example of an extended-release proprietary product made using a HPMC hydrophilic matrix is the extended-release morphine sulfate tablet (roxatidine; see Martindale 33rd Ed., p.2014.4).

Bilayer tablets can be prepared containing one or more compounds of the invention and formulations, with one layer containing unconjugated drug as an immediate release layer and the other layer containing drug embedded in a hydrophilic matrix as an extended release layer . A three-layer tablet can also be prepared in a similar manner, with the two outer layers containing the drug as the immediate release layer. Some commercially available tablets contain an extended-release drug in an inner core and an encapsulating shell that contains a rapid-release drug.

The invention also provides complexes of active ingredients, such as one or more compounds and formulations of the invention, with ion exchange resins, which complexes can be tabletted, encapsulated or suspended in an aqueous carrier. The release of the active agent is dependent on the local pH and electrolyte concentration, and the choice of such an ion exchange resin is preferred over a resin that releases the active agent at the corresponding location in the alimentary canal. The invention also provides delivery devices incorporating the complex. For example, a sustained-release dosage form of triethylenetetramine can be obtained by combining triethylenetetramine with an anion exchange resin. The triethylenetetramine solution can be passed through the ion exchange resin column to form complexes by replacing H ₃ O ⁺ ions. The resin-triethylenetetramine complex is then washed and can be tabletted, encapsulated or suspended in an aqueous carrier. The release of triethylenetetramine depends on the pH and electrolyte concentration of the gastrointestinal fluid. The release is greater in the acidic gastric juice than in the small intestinal fluid in a less acidic environment. Another example of an extended release formulation of this type is provided by hydrocodone sulfonated divinylbenzene-vinylbenzene copolymer and chorpheniramine polistirex suspension (Medeva; Tussionex Pennkinetic Extended Release Suspension, see Martindale 33rd Ed., p.2145.2), and provided by phentermine resin capsules (Pharmanex; Ionamin Capsules see Martindale 33rd Ed., p. 1916.1). Such a resin-triethylenetetramine active agent system may additionally incorporate polymeric barrier coating and beading techniques in addition to ion exchange techniques. The initial dose is from the uncoated portion, the remainder from the coated beads. The coating does not dissolve and drug release can be extended over 12 hours due to ion exchange. Drugs comprising particles are small and can be suspended to form liquid dosage forms as well as solid dosage forms which can have extended release properties. Such formulations may also be suitable for administration in depot dosage forms such as intramuscular injection.

The present invention also provides a process for the preparation of a sustained release formulation of one or more copper chelating agents, such as triethylenetetramine or a salt thereof, by microencapsulation. The microencapsulated formulations are useful in the treatment of humans and other mammals in need of copper chelation therapy. Microencapsulation is the process by which solids, liquids and even gases can be encapsulated in tiny particles by forming a film coating on the surface of the substance. The microencapsulation method can be found in U.S. Patent Nos. 3,488,418; 3,391,416 and 3,155,590. Gelatin (BP, USP) can generally be used as a coating wall forming material for microencapsulated formulations, while synthetic polymers such as polyvinyl alcohol (USP), ethylcellulose (BP, USP), polyvinyl chloride, and other materials can also be applied (see Zentner, G.M., Rork, G.S. & Himmelstein, K.J. Osmotic flow through controlled porosity films: an approach to delivery of water soluble compounds. J Controlled Release 2: 217-229 (1985); Fites, A.L., Banker, G.S. & , V.F.Controlled drug release through polymeric films.J PharmSci59: 610-613(1970); Samuelov, Y., Donbrow, M. & Friedman, M.Sustained release of drugs from ethylcellulose-polyethylene glycol Pharm leof kinetics J drugase. Sci 68:325-329 (1979)).

Encapsulation begins with the dissolution of the coating wall material, such as gelatin, in water. One or more copper chelating agents such as triethylenetetramine or its salts are then added and the two-phase mixture is stirred thoroughly. When the encapsulated material has reached the desired particle size, a solution of another material, which may be gum arabic (BP, USP), is added. Additional materials are selected for their ability to condense the gelatin (polymer) into tiny droplets. These droplets (agglomerates) then form a thin film or coating on the solid triethylenetetramine particles, resulting in minimal interfacial tension of residual water or solvent in the coating wall material, allowing the formation of a solid continuous (See Ansel, H.C., Allen, L.V. and Popovich, N.G. Pharmaceutical Dosage Forms and Drug Delivery Systems, 7th Ed., Lippincott 1999, p.233). The final dry microcapsules are free-flowing, discrete coated particles. The coating wall material is typically 2-20% (w/w) relative to the total granule weight. The coated granules are then blended with tablet excipients to form tablets of the appropriate dosage. The release rate of different triethylenetetramine can be obtained by changing the ratio of core-coating wall, the polymer used for coating, or the method of microencapsulation (for example, referring to Yazici, E., Oner, L. , Kas, H.S. & Hincal, A.A. Phenytoin sodium microspheres: bench scaleformulation, process characterization and release kinetics. Pharmaceut DevTechnol 1996; 1: 175-183).

One advantage of microencapsulation is that the administered dose of one or more copper chelating agents such as triethylenetetramine or its salts is divided into multiple small dosage units, which can be widely distributed in the gastrointestinal tract, by reducing the local drug concentration to increase drug absorption (see Yazici et al., supra). An example of a commercially available microencapsulated extended release dosage form is potassium chloride (Micro-KExten-caps, Wyeth-Ayerst, Martindale 33rd Ed., p1968.1). Others include those in which the drug is incorporated into polymeric colloidal particles or microcapsules (microparticles, microspheres, or nanoparticles) to form depot and matrix devices (see Douglas, S.J., et al., "Nanoparticles in drug delivery," C.R.C. Crit Rev Therapy Drug Carrier Syst 3: 233-261 (1987); Oppenheim, R.C., "Solid colloidal drug delivery systems: nanoparticles." Int J Pharm 8: 217-234 (1981); Higuchi, T. "Mechanism of sustained action medication: theoretical analysis of rate of release of soliddrugs dispersed in solidmatrices." J Pharm Sci 52:1145-1149(1963)).

The invention also includes re-action tablets containing one or more copper chelating agents such as triethylenetetramine or salts thereof. Further included are methods of preparing sustained release dosage forms of one or more copper chelating agents, such as triethylenetetramine or salts thereof, suitable for use in the treatment of humans or other mammals by combining triethylenetetramine with reproducible action available in tablets. Tablets are prepared so that an initial dose of drug is released rapidly followed by a second dose. The tablet has an immediate release dosage layer as an outer shell or coating, a second dosage layer is in the inner core of the tablet separated by a coating barrier with a permeation retarding effect. Typically, the drug in the inner core is exposed to body fluids and released 4-6 hours after administration. Examples of products of this type are confirmed to exist by Repetabs (Schering Inc.). Repeat-action dosage forms are suitable for the administration of one or more copper chelating agents such as triethylenetetramine or its salts, for conditions including but not limited to chronic diseases such as heart failure, diabetic heart disease, acute coronary syndrome , hypertensive heart disease, ischemic heart disease, coronary heart disease, peripheral arterial disease, or any form of cancer. This form of drug release is particularly suitable for the release of triethylenetetramine due to its rapid absorption and excretion.

The present invention also includes delayed release oral dosage forms comprising one or more copper chelating agents such as triethylenetetramine or salts thereof. The release of an oral dosage form of one or more copper chelating agents such as triethylenetetramine or its salts may be intentionally delayed until the drug reaches the intestine by, for example, an enteric coating. Enteric coatings by themselves are not an effective means of releasing copper chelators such as triethylenetetramine or its salts, including triethylenetetramine dihydrochloride, as such coating systems do not provide long-lasting therapy after drug release Effect. Enteric coatings need to dissolve or disintegrate in an alkaline environment. The presence of food increases the pH in the stomach. Therefore, ingestion of enteric-coated triethylenetetramine dihydrochloride with food or in the presence of food in the stomach can lead to loss of drug dose and occurrence of undesired side effects. Moreover, it has been shown that TETH can cause gastrointestinal side effects, so there is a need for a drug delivery system that can deliver controlled doses of TET or other pharmaceuticals. Acceptable release of the salt of triethylenetetramine over an extended period of time in a predetermined manner.

Enteric coatings may also be used in the present invention when the formulation or device is delivered in combination with one or more of the other formulations described herein. The advantage of this release form is that it reduces the gastrointestinal irritation that triethylenetetramine may cause in some subjects. Enteric coatings can be time-dependent, pH-dependent, where they disintegrate in the less acidic intestine and are eroded by moisture over time during intestinal transit, or enzyme-dependent, where Under the action of Dao enzyme, it catalyzes hydrolysis and deteriorates (see "Modifying the release properties of Eudragit L30D," by Muhammad, N.A., et al., Drug Dev Ind Pharm. 17: 2497-2509 (1991)). A variety of agents for enteric coating of tablets and capsules are well known to those skilled in the art, including fats including triglycerides, fatty acids, waxes, shellac and cellulose acetate phthalate, more enteric Dissolving formulations can be found in USP.

The present invention also includes drug delivery devices for membrane control systems comprising one or more copper chelating agents such as triethylenetetramine or salts thereof. Such devices include a rate controlling membrane surrounding a drug depot. After oral administration, the film becomes gradually liquid permeable but does not erode or swell. The drug depot can be made as a traditional tablet, or as a microparticle pellet comprising multiple units, which does not swell upon contact with liquid. The core can be dissolved without changing the internal osmotic pressure, thus avoiding damage to the membrane, and typically contains a 60:40 mixture of lactulose:microcrystalline cellulose (w/w). Drug release occurs through a two-phase process involving diffusion of aqueous fluid into the matrix followed by drug diffusion out of the matrix. Multi-unit thin film control systems typically contain multiple separate units. May contain discrete spherical beads coated with a rate controlling film and may be encapsulated in a hard gelatin shell (examples of such formulations include Contac 400; Martindale 33rd Ed., 1790.1 and Ferrous fumarate formulation Feospan; Martindale 33rd Ed., p.1859.4). Alternatively, multi-unit film control systems can be compressed into tablets (eg, racemic isoproterenol Suscard; Martindale 33rd Ed., p. 2115.1). Another way of using this technology involves a device in which the drug is coated on inert sugar spheres, prepared by extrusion spheronization using conventional matrix systems. Advantages of this system include increased gastrointestinal transit rates without loss of drug dose. The device is also ideally capable of delivering more than one drug at a time.

Preferred oral delivery refers to the sustained release form of one or more compounds and formulations of the invention as a matrix structure in the form of a film-coated spheroid containing as active ingredient one or more Various copper chelating agents such as triethylenetetramine or its salts such as triethylenetetramine dihydrochloride, and water-insoluble spheroidizing agents. The term "spheroid" refers to pharmaceutically spherical particles, usually 0.01 mm to 4 mm in diameter. The spheroidizing agent can be any pharmaceutically acceptable material, which can be formed into a spherical shape when mixed with the active ingredient. Microcrystalline cellulose is preferred. Suitable microcrystalline celluloses include, for example, Avicel PH 101 (trade mark, FMC Corporation). According to a preferred aspect of the invention, the film-coated pellets comprise 70% to 99% by weight, especially 80% to 95% by weight, of a spheroidizing agent, especially microcrystalline cellulose. In addition to the active agent and spheronizing agent, the pellets may also contain a binder. Suitable binders, such as low viscosity water soluble polymers, are well known to those skilled in the art. A suitable binder is in particular polyvinylpyrrolidone of various degrees of polymerization. However, low alkyl celluloses with water soluble hydroxyl groups, such as hydroxypropyl cellulose, are preferred. Additionally (or alternatively), the pellets may comprise water insoluble polymers, especially acrylate polymers, acrylate copolymers, such as methacrylate-ethyl acrylate copolymer or ethyl cellulose. Other thickeners or binders include: Fats, including vegetable oils (cottonseed, sesame, and peanut oils) and their derivatives (hydrogenated oils such as hydrogenated castor oil, glycerin, waxy substances such as natural carnauba wax or natural beeswax , synthetic waxes such as cetyl ester wax, amphoteric substances such as ethylene oxide polymers (high molecular weight polyoxyethylene glycol, molecular weight 4000-100000) or copolymers of ethylene oxide and propylene oxide (poloxamer ), cellulosic substances (cellulose, hydroxypropylmethylcellulose, hydroxypropylcellulose, semi-synthetic derivatives of hydroxymethylcellulose high-molecular-weight high-viscosity resins) or any other polysaccharide such as alginic acid, Polymer substances such as acrylic polymers (such as carbomer), and inorganic substances such as colloidal silica and bentonite.

Suitable diluents for the active ingredient in pellets, pellets or cores are, for example, microcrystalline cellulose, lactose, dicalcium phosphate, calcium carbonate, calcium sulfate, sucrose, dextrose, dextrin, dextrose, dicalcium phosphate Hydrate, kaolin, magnesium carbonate, magnesium oxide, maltodextrin, cellulose, microcrystalline cellulose, sorbitol, starch, pregelatinized starch, talc, tricalcium phosphate and lactose. Suitable lubricants are, for example, magnesium stearate and sodium stearyl fumarate. Suitable binders are eg hydroxypropylmethylcellulose, polyvidone and methylcellulose.

Suitable binders may include: Gum Arabic, Gum Tragacanth, Guar Gum, Alginic Acid, Sodium Alginate, Sodium Carboxymethylcellulose, Dextrin, Gelatin, Hydroxyethylcellulose, Hydroxypropylcellulose , Liquid Glucose, Magnesium and Aluminum. Suitable disintegrants are starch, sodium starch glycolate, crospovidone and croscarmellose sodium. Suitable surfactants are Poloxamer 188(R), Tween 80 and sodium lauryl sulfate. A suitable glidant is talc colloidal anhydrous silicon dioxide. Suitable lubricants that may be used may be slip agents such as anhydrous silicate, magnesium trisilicate, magnesium silicate, cellulose, starch, talc or tricalcium phosphate, or lubricants such as stearic acid calcium, hydrogenated vegetable oil, paraffin, magnesium stearate, macrogol, sodium benzoate, sodium lauryl sulfate, fumaric acid, stearic acid or zinc stearate and talc). A suitable water soluble polymer is PEG with a molecular weight in the range 1000-6000.

Tablets, pellets, pellets or cores themselves for delayed release contain, in addition to fillers and binders, other adjuvants, in particular lubricants and antiadherents, and disintegrants. Examples of lubricants and anti-adherents are higher fatty acids and their alkali metal and alkaline earth metal salts, such as calcium stearate. Suitable disintegrants are especially those which are chemically inert. The disintegrants are preferably cross-linked polyvinylpyrrolidone, cross-linked sodium carboxymethylcellulose and sodium starch glycolate.

The oral dosage unit preferably delivers more than 50% of the copper chelator such as triethylenetetramine dihydrochloride within 12 hours at a pH < 6.5 in a controlled manner for in vivo and in vitro dissolution. Other formulations and dosage forms are described below.

Further embodiments of the invention include dosage forms in which one or more copper chelating agents such as triethylenetetramine or salts thereof are combined with a transdermal delivery system, such as those described in Transdermal Drug Delivery Systems, Chapter 10. In: Ansel, H.C., Allen, L.V. and Popovich, N.G. Pharmaceutical Dosage Forms and Drug Delivery Systems, 7th Ed., Lippincott 1999, pp.263-278. The delivery system for transdermal drug delivery is conducive to the therapeutic dose of the drug entering the systemic circulation through the skin to exert its effect, which can be referred to (Stoughton, R.D. Percutaneous absorption. Toxicol Appl Pharmacol 7: 1-8 (1965)). Evidence of transdermal drug absorption can be determined by measuring blood drug levels, measuring drug excretion and/or metabolites in urine following administration of the subject's clinical response. For transdermal drug delivery, it is considered ideal if the drug penetrates the skin into the blood without accumulating in the skin layer (Black, C.D., "Transdermal drug delivery systems," U.S. Pharm 1:49 (1982)). Pharmaceutical formulations suitable for transdermal delivery are well known to those skilled in the art and are described in several references, eg Ansel et al. (supra). Known methods of enhancing drug delivery via the transdermal route include chemical transdermal enhancers, which increase the permeability of the skin by reversibly damaging or otherwise altering the physicochemical properties of the stratum corneum to reduce its resistance to drug diffusion (See Shah, V.P., Peck, C.C. & Williams, R.L. Skin penetration enhancement: clinical pharmacological and regulatory considerations. In: Walters, K.A. & Hadgraft, J. (Eds.) Pharmaceutical skin penetration enhancement. New York: Dekker, 1993 ). Other alterations that are effective are increased hydration of the liver cuticle by solvent activity or denaturation, and/or changes in lipid and lipoprotein structure in intercellular channels (see Walters K.A., "Percutaneous absorption and transdermal therapy, "Pharm Tech 10:30-42 (1986)). The skin penetration enhancer of triethylenetetramine formulations suitable for transdermal drug delivery systems can be selected from the following listed materials: acetone, laurocapram (Laurocapram, 1-n-dodecyl nitrogen heterocycle heptan-2-one), dimethylacetamide, dimethylformamide, dimethylsulfoxide, ethanol, oleic acid, polyethylene glycol, propylene glycol, and sodium lauryl sulfate. Further given skin penetration enhancers can be found in conventional publications by those skilled in the art (see Osborne, D.W., & Henke, J.J., "Skin penetration enhancers cited in the technical literature," Pharm Tech 21:50-66( 1997); Rolf, D., "Chemical and physical methods of enhancing transdermal drug delivery," Pharm Tech 12:130-139 (1988)).

In addition to chemical means, physical means can also be used to enhance the transdermal release and penetration of the compounds and formulations of the invention. These include iontophoresis and sonophoresis. Iontophoresis involves the application of an electric field to cause drug release of chemicals across the skin membrane. The method is applicable to the release of many drugs. Accordingly, another embodiment of the present invention comprises one or more copper chelating agents such as triethylenetetramine or its salts into suitable formulations using iontophoresis or sonophoresis. One or more copper chelating agents such as triethylenetetramine or its salts are prepared using iontophoresis or sonophoresis. Suitable formulations may be in the form of gels, creams or lotions. Transdermal drug delivery can be applied to other systems such as monolithic drug delivery systems, drug osmoadhesive delivery systems (such as the Latitude ^™ drug attachment system from 3M), active drug release devices, and membrane control systems. The monolithic system incorporates an active agent matrix comprising a polymeric material into which the active agent can be dispersed between its front and back layers. The osmotic drug delivery system comprises an adhesive polymer in which one or more compounds of the present invention and formulations and excipients are incorporated. Active transport devices are combined with active agent reservoirs, usually in liquids or gels, and membranes can be used for rate control as the motive force for propelling the active agent across the membrane. Membrane-controlled transdermal delivery systems typically include a reservoir of active agent in liquid or gel form, a membrane as a rate controlling, support, attachment and/or protective layer. Transdermal delivery dosage forms include those active ingredients substituted with triethylenetetramine, preferably triethylenetetramine dihydrochloride or other pharmaceutically acceptable salts, see US Patent No. 6,193,996 for examples in transdermal delivery systems and 6,262,121.

Topical administration of the compounds and formulations of this invention may be prepared as admixtures or other pharmaceutical formulations for application in a variety of ways including, but not limited to, lotions, creams, gels, sticks, sprays, ointments, and pastes. agent. These product types can include a variety of formulation types including, but not limited to, solutions, emulsions, gels, solid doses, and liposomes. If the topical composition is formulated as an aerosol and sprayed onto the skin, a propellant may be added to the solution composition. Conventional suitable propellants can be used. See U.S. Patent Nos. 5,602,125, 6,426,362 and 6,420,411 for examples of topical compositions.

The invention also includes sustained release dosage forms, which may be various oral dosage form variants adapted for suppository or other parenteral use. When administered rectally in the form of suppositories, these compositions may be prepared by admixing one or more of the compounds and formulations of this invention with suitable non-irritating excipients, such as cocoa butter, which is solid at ordinary temperatures, Synthetic glycerides or polyethylene glycols, which liquefy and/or dissolve in the rectum. Suppositories are usually solid dosage forms for insertion into cavities in the body, including the rectum, vagina, and occasionally the urethra, and may be long-acting or slow-release. Suppositories include a base including, but not limited to, materials such as alginic acid, which prolong the release of the pharmaceutically active ingredient for several hours (5-7 hours). These matrices can be classified by nature into two broad categories and a third mixed group: 1) fatty or oily matrices, 2) water-soluble or water-miscible matrices, and 3) mixed-type matrices, usually lipophilic and hydrophilic Mixture of substances. Fatty or oily bases including hydrogenated fatty acids of vegetable oils such as palm kernel oil and cottonseed oil, fatty substances containing glycerol and high molecular weight fatty acids such as palmitic and stearic acids, and cocoa butter can also be used, in which phenol and chloral hydrate are reduced Depending on the melting point of cocoa butter, solidifying agents such as cetyl esters wax (about 20%) or beeswax (about 4%) can be added to maintain the solid form of the suppository. Other bases include commercially available products such as lipid antiacid suppository bases (triglycerides and self-emulsifying glyceryl monostearate and poloxyl stearate from palm, palm kernel, and coconut oils). , Wecobee and Witepsol substrates. The water soluble base is usually glycerinated gelatin and the water-miscible base is usually polyethylene glycol. Hybrid bases include oleaginous and water soluble or water miscible materials. An example of such a base is polyoxyethylene 40 stearate and polyoxyethylene glycol and free ethylene glycol.

Transmucosal delivery of the compounds and formulations of the invention may employ any mucosal membrane commonly used for administration to intranasal, buccal, vaginal and rectal tissues.

Dosages of the invention suitable for nasal administration may be administered in liquid form, eg nasal spray, nasal drops, or in aerosol form via a nebuliser, including aqueous or oily solutions of the active ingredient. Formulations for nasal administration, wherein the carrier is a solid, comprise a coarse powder having a particle size, for example less than 100 microns in diameter, preferably less than 50 microns, which is administered as a nasal inhalation medicament, i.e. a container containing the powder Administer by inhaling quickly close to the nasal cavity. Solution compositions can be administered using a built-in gas spray, and the nebulized solution can be inhaled directly from the nebulizer device, which can be attached to a face mask, tampon or intermittent pressurized respirator. Solution, suspension or powder compositions can be administered orally or nasally, by suitable means for drug delivery. Formulations may be prepared as aqueous solutions such as saline solutions, solutions using benzyl alcohol or other suitable preservatives, absorption enhancers to increase bioavailability, fluorocarbons and/or other known soluble or dispersible substances.

The present invention also provides extended release formulations suitable for parenteral administration comprising one or more copper chelating agents such as triethylenetetramine or salts thereof. Prolonged drug activity for parenteral administration can be achieved in a variety of ways, including: crystalline or amorphous drug forms with prolonged dissolution properties; slow dissolution of chemical complexes of the drug; in slowly absorbed carriers or vehicles ( solution or suspension of drug as oleaginous); increased particle size of drug in suspension; or slow erosion of injected drug microspheres (see for example Friess, W., Lee, G. and Groves, M. J. Insoluble collagen matrices for prolonged delivery of proteins. Pharmaceut Dev Technol 1: 185-193 (1996)). The duration of various forms of eg insulin is based in part on its physical form (amorphous or crystalline), the formation of complexes with additional agents, and its dosage form (solution or suspension).

Copper chelators need to be formulated as stable, safe pharmaceutical compositions for administration to patients. The copper chelator is a triethylenetetramine active agent. The composition can be prepared according to conventional methods by dissolving or suspending an amount of triethylenetetramine active agent in a diluent. The amount is 0.1-1000 mg of triethylenetetramine active agent per ml of diluent. Acetate, phosphate, citrate or glutamate buffers may be added to bring the pH of the final composition between 5.0-9.5; optionally carbohydrate or polyol regulators, and preservatives selected from m-cresol, benzyl alcohol, methyl, ethyl, propyl and butyl substituted parabens and phenol. Sufficient water for injection is used to obtain the required solution concentration. Additional regulators such as sodium chloride and other auxiliaries may also be present, if desired. The adjuvant must be able to maintain all the properties of the triethylenetetramine active agent.

The terms buffering agent and buffer solution refer to the hydrogen ion concentration or pH value, and refer to the ability of a system, especially an aqueous solution, to resist changes in pH value after adding an acid or base or diluting with a solvent. The characteristic of a buffer solution is that it can withstand small changes in pH value after adding an acid or base, and it often exists in the form of a weak acid and its salt, or a weak base and its salt. An example of this system is acetic acid and sodium acetate. The change in pH value is very small, as long as the addition of hydrogen ions is not too much, the buffer system can offset it.

The stability of the parenteral formulations of the present invention can be enhanced by maintaining the pH in the range of about 5.0-9.5. Other pH ranges include, for example, 5.5-9.0, or 6.0-8.5, or 6.5-8.0, or 7.0 to 7.5.

The buffer used in the present invention is selected from any one of the following, such as acetate buffer, phosphate buffer or glutamic acid buffer, most preferably phosphate buffer.

Carriers or excipients can be used to facilitate administration. Examples of carriers or excipients include calcium carbonate, calcium phosphate, various sugars such as lactose, glucose or sucrose, or various types of starch, cellulose derivatives, gelatin, polyethylene glycol and physiologically compatible solvents.

The formulations according to the invention may include stabilizers, but it is emphasized that they are not required. However, if included, stabilizers useful in the present invention are carbohydrates or polyols. Polyols include sorbitol, mannitol, glycerol and polyethylene glycols (PEGs). Carbohydrates are mannose, ribose, trehalose, maltose, fiber sugar, lactose, galactose, arabinose or lactose.

Suitable stabilizers include, for example, polyalcohols such as sorbitol, mannitol, inositol, glycerol, xylitol and polypropylene/ethylene glycol copolymers and various molecular weights of 200, 400, 1450, 3350, 4000, 6000 and 8000 polyethylene glycol (PEG).

The United States Pharmacopeia (USP) specifies that bacteriostatic or fungistatic concentrations of antimicrobial agents must be added to containers of multi-dose formulations. It must be used in a sufficient concentration to prevent the retraction of part of the contents when using a hypodermic needle, or the proliferation of microorganisms brought about by other methods of administration such as pen injection. Antimicrobial agents are evaluated to ensure compatibility with all other ingredients, and their activity is evaluated to identify specific agents that are effective in one formulation but not in another. It is easy to determine that a particular agent is effective in one formulation but not in another.

In common pharmaceutical practice, preservatives prevent or inhibit the growth of microorganisms and can be added to pharmaceutical formulations where this is required, in order to avoid deterioration of the formulation by the influence of microorganisms. But although the consumption of preservative is not very much, still can influence the total stability of triethylenetetramine active agent. Therefore, the choice is more difficult.

Among the present invention, the consumption scope of preservative is at 0.005-1.0% (w/v), and preferably for each kind of preservative, the consumption when alone or combined application is: benzyl alcohol (0.1-1.0%), m-cresol (0.1- 0.6%), or phenol (0.1-0.8%), or a combination of methyl (0.05-0.25%) and ethyl or propyl or butyl (0.005%-0.03%) parabens. Parabens are lower alkyl esters of p-hydroxybenzoic acid.

A detailed description of each preservative can be found in "Remington's Pharmaceutical Sciences" and "Pharmaceutical Dosage Forms: Parenteral Medications, Vol. 1, 1992, Avis et al.". For these purposes, the salts of triethylenetetramine dihydrochloride in the crystalline form can be used for parenteral administration (including subcutaneous injection, intravenous injection, intramuscular injection, intradermal injection or infusion techniques) or by inhalation Nebulization administration administers formulations in dosage units comprising conventional non-toxic pharmaceutically acceptable carriers, adjuvants and vehicles.

There may also be a need to add sodium chloride or other salts to adjust the tonicity of the pharmaceutical formulation, depending on the tonicity modifier chosen. However, this is an optional component and depends on the particular formulation chosen. Formulations for parenteral administration must be isotonic or approximately isotonic or they can cause significant irritation and pain in the area of administration.

The desired isotonicity can be achieved by adding sodium chloride or other pharmaceutically acceptable agents, such as dextrose, boric acid, sodium tartrate, propylene glycol, polyols (eg, mannitol and sorbitol), or other inorganic or organic solutes. Typically, compositions are isotonic with the blood of the subject.

Formulations for parenteral administration can, if desired, be thickened with a thickening agent such as methylcellulose. Formulations can be prepared as oil-in-water or water-in-oil emulsions. Any pharmaceutically acceptable emulsifying agent can be used, such as gum arabic powder, nonionic or ionic surfactants.

Suitable dispersing or suspending agents may also be added, including aqueous suspending agents such as synthetic and natural gums, i.e. tragacanth, acacia, alginates, dextran, sodium carboxymethylcellulose, methylcellulose, Polyvinylpyrrolidone or gelatin.

The most important carrier for products for parenteral administration is water. Appropriate amounts of water must be distilled or reverse osmosis for parenteral administration. Only in this way can various solid, liquid and gaseous pollutants be fully separated from the water. Water for injection is a preferred aqueous carrier that can be used in the pharmaceutical formulations of the present invention. Water can be purged with nitrogen to remove oxygen or oxygen free radicals from the water.

The formulations for parenteral administration according to the invention may also contain other ingredients. These additional ingredients may include wetting agents, oils (such as vegetable oils, such as sesame oil, peanut oil, or olive oil), analgesics, emulsifiers, antioxidants, bulking agents, tonicity regulators, metal ions, oily carriers, proteins (such as human serum albumin, gelatin or protein) and zwitterions (eg amino acids like betaine, taurine, arginine, glycine, lysine and histidine). Such additional constituents must of course not adversely affect the overall stability of the pharmaceutical formulation according to the invention.

The container is also an integral part of the injection, and can be considered a component, since no container is completely insoluble or does not have any effect on the liquid in it, especially when the liquid is aqueous. Therefore, especially the choice of the container for injection must be based on the composition, solution and treatment to be carried out.

To allow the hypodermic needle to enter the multi-dose vial and maintain the seal when the needle is withdrawn, each vial is sealed with an aluminum ring with a rubber stopper.

Glass vial stoppers such as West 4416/50, 4416/50 (Teflon faced) and 4406/40, Abbott 5139 or any other equivalent stoppers can be used for sealing the dosage vials of the present invention. These stoppers are subjected to complete cork testing, i.e. the stoppers are able to withstand at least 100 injections.

Each ingredient of the pharmaceutical formulations described above is well known to those skilled in the art and can be found in "Pharmaceutical Dosage Forms": Parenteral Medications, Vol. 1, 2nd ed., Avis et al. Ed., Mercel Dekker, New York, N.Y. 1992, which is incorporated herein by reference in its entirety.

The preparation process of the above formulation includes the steps of compounding, sterile filtration and filling. The compounding process may, for example, involve dissolving in a specific order, such as preservative first, followed by stabilizer/tonicity modifier, buffer, and then TET active agent, or dissolving all components simultaneously to form a parenteral formulations for oral administration. An example of one method of preparing a formulation for parenteral administration is to dissolve the active form of triethylenetetramine, such as triethylenetetramine dihydrochloride, in water and dilute the resulting mixture into phosphate-buffered saline 154mM.

Alternatively, parenteral formulations of the present invention can be prepared by admixing the ingredients after the conventional procedure. For example, selected components can be mixed in a mixer or other standard apparatus to obtain a concentrated mixture, and water, thickeners, buffers, 5% human serum albumin, or additional tonicity-adjusting solutes can be added to obtain the final mixture. concentration and viscosity.

Alternatively, the triethylenetetramine active agent can be packaged in the form of dry solid and/or powder, and reconstituted by adding a solvent during use to obtain the reconstituted formulation for parenteral administration of the present invention.

The preparation process also includes suitable sterilization processes. Typical sterilization processes include filtration, steam treatment (moist heat), dry heat treatment, gas treatment (e.g. ethylene oxide, formaldehyde, chlorine dioxide, propylene oxide, β-propiolactone, ozone, nitrochloroform, peroxy Acetate methyl bromide, etc.), radiation treatment and aseptic treatment.

Suitable routes of parenteral administration include intramuscular injection, intravenous injection, subcutaneous injection, intradermal injection, intraarticular injection, intrathecal injection and the like. The preferred route of administration is subcutaneous injection. Mucosal release administration is also possible. The dosage and dosage regimen will depend on the weight and health of the subject.

Parenteral routes of administration include intravenous, intramuscular, intraperitoneal, transdermal, and subcutaneous.

In addition to the above-mentioned ways of prolonging the drug effect, it can also be achieved by controlling the release rate and duration of the drug, for example, by using a mechanically controlled drug infusion pump.

A pharmaceutically acceptable active agent, such as one or more copper chelating agents such as triethylenetetramine or a salt thereof such as triethylenetetramine dihydrochloride, may be administered as a depot injection to obtain Sustained release of active agent. The active ingredient can be compressed into pellets or cylinders for subcutaneous or intramuscular implantation. The pellets or cylinders can be coated with suitable biodegradable polymeric materials to obtain the desired release profile. The active ingredient can also be pelletized. Pellets of the active agent using biodegradable polymers can be designed at the desired release rate to obtain the desired release profile. Alternatively, injectable depot forms can be made by forming microencapsule matrices of the major substance in biodegradable polymers such as polylactide-polyglycolide. Depending upon the ratio of drug to polymer, and the nature of the particular polymer employed, the rate of drug release is controlled. Examples of other biodegradable polymers include polyorthoesters and polyanhydrides. Injectable depot formulations can also be prepared by entrapping the drug in liposomes, examples of which include unilamellar vesicles, unilamellar bullous liposomes and multilamellar liposomes. Liposomes can be formed from a variety of phospholipids, such as cholesterol, stearylamide or phosphatidylcholines. Depot injectable drug formulations can also be formed by entrapping the drug in microemulsions which are compatible with body tissues. Examples thereof can be found in U.S. Patent Application Nos. 6,410,041 and 6,362,190.

The present invention also provides infusion drug delivery formulations and devices, including but not limited to implantable infusion devices. Implantable infusion devices may be obtained using inert materials such as the biodegradable polymers mentioned above or synthetic silicones such as silicone rubber or other Dow-Corning polymers. Polymers can be loaded with active agents and excipients. Implantable infusion devices may also be coated or partially coated, wherein the coating comprises polymers loaded with active agent and excipients. Such an implantable infusion device can also be prepared according to the method of U.S. Patent No. 6,309,380 by encapsulating the device with a liquid or gel solution containing an in vivo compatible biodegradable or bioabsorbable or bioerodible A coating wherein the solution contains a polymer comprising the desired dose of active ingredient and excipients. The solution adheres to form a film on the device, forming an implantable drug-releasing medical device.

Implantable infusion devices can also be prepared in situ containing the active agent in a solid matrix, see U.S. Patent No. 6,120,789, which is incorporated herein by reference in its entirety. Implantable infusion devices can be passive transport or active transport. An active implantable infusion device may comprise a depot of active agent, means for releasing the active agent from the depot, such as a permeable membrane, and drive means for urging the active agent out of the depot. Such an active implantable infusion device can be activated by an external signal, as described in WO 02/45779, wherein the implantable infusion device contains a system for releasing the active agent, including an external active unit, which can be activated by A user action required to activate the implantable infusion device includes a controller for inhibiting the need until the blockade interval expires. Examples of active implantable infusion devices include implantable drug pumps. Implantable drug pumps include, for example, miniature, computerized, programmable, refillable drug delivery systems with attached catheters that can be inserted into target organs, usually the spinal cord or blood vessels.参见 Medtronic Inc.Publications：UC9603124 ENNP-2687，1997；UC199503941b EN NP-2347182577-101，2000；UC199801017a EN NP3273a182600-101，2000；UC200002512 EN NP4050，2000；UC199900546b EN NP-3678 EN，2000.Minneapolis，Minn： Medtronic Inc; 1997-2000. Many pumps have two parts: a part that injects the drug and a part that connects directly to the catheter for bolus injection or analysis of the fluid in the catheter. Implantable drug infusion pumps (SynchroMed EL and Synchromed Programmable Pumps; Medtronic) are available for long-term intrathecal morphine sulfate infusion for chronic refractory pain; intravascular infusion of fluorouracil-deoxynucleoside for in situ cancer or metastases; intrathecal injections (baclofen injections) for severe spasticity; long-term epidural infusions of morphine sulfate for chronic refractory pain; long-term intravascular infusions of doxorubicin, cisplatin, or methotrexate Treatment of metastatic cancer; and long-term intravenous infusion of clindamycin for osteomyelitis. Such pumps may also be used for chronic infusion of one or more copper chelating agents such as triethylenetetramine or salts thereof at desired doses or steady state administration. A typical form of implantable drug infusion pump (Synchromed EL programmable pump; Medtronic) is titanium-coated and roughly disc-shaped, 85.2 mm in diameter, 22.86 mm in thickness, weighs 185 g, and contains 10 ml of drug Storage, depending on usage, lithium thionyl chloride battery life is 6-7 years. A downloadable memory store includes programmed drug release parameters and calculated drug remaining which can be compared to actual drug remaining to approximate pump function accuracy, but actual pump function over time is not recorded. The pump is usually implanted in the left or right abdominal wall. Other pumps that can be used in the present invention include portable disposable infusion pumps (PDIPs). In addition, implantable infusion devices can employ liposome delivery systems, such as small unilamellar vesicular liposomes, unilamellar bullous liposomes, and multilamellar liposomes, which can be formed by various phospholipid substances, such as Cholesterol, stearylamide or phosphatidylcholine.

The present invention also includes delayed release ophthalmic pharmaceutical formulations which include one or more copper chelating agents such as triethylenetetramine or salts thereof. Retinal artery disease leading to plasma leakage and eventually diabetic retinopathy is the leading cause of visual impairment and blindness in diabetic patients. Triethylenetetramine is effective in the treatment of diabetic arterial disease. The invention provides an ophthalmic preparation of triethylenetetramine, which is suitable for treating retinal artery disease caused by diabetes. The administration forms a local high concentration of the drug, which is suitable for the treatment of diabetic retinal artery disease and diabetic retinopathy.

One problem with the use of ophthalmic solutions is the rapid loss of the drug by blinking and tears. Up to 80% of the drug dose is lost within 5 minutes through the tears and nasolacrimal duct. Prolonged treatment can be accomplished by increasing the contact time of the drug with the corneal surface. This may require increasing the viscosity of the solution; applying a slowly dissolving ophthalmic suspension of drug particles; applying a slowly dispersing ophthalmic ointment; or applying an ophthalmic implant. The formulation of one or more copper chelating agents such as triethylenetetramine or its salts for human ophthalmology may use synthetic high molecular weight cross-linked polymers such as those of acrylic polymers (e.g. carbopol 940) or glue Gelrite (Ref. Merck Index 12th Ed., 4389), a substance that forms a gel in contact with the precorneal tear film (eg as used in Timoptic-XE by Merck, Inc.).

Further embodiments of the invention include delayed release ophthalmic formulations comprising triethylenetetramine implants, such as the OCUSERT system (Alza Inc.). Typically, such implants are oval in shape and measure approximately 13.4mm x 5.4mm x 0.3mm (thickness). The implant is flexible, has a drug-containing core, and is coated with a hydrophobic vinyl acetate copolymer membrane that allows the drug to diffuse at a constant rate. The white edge of the device contains white titanium dioxide, which is a distinguishable inert substance. Drug diffusion rate can be controlled by polymer composition, film thickness, and drug solubility. In the first few hours after implantation, the drug release is faster than after that to obtain the starting drug concentration level. Drug-containing implants can be placed in the conjunctival sac and typically have a drug release period of 7 days in the treatment of diabetic retinal disease. Another type of ophthalmic implant is a rod-shaped, water-soluble structure consisting of hydroxypropylcellulose with triethylenetetramine in it. The implant is placed in the conjunctival fornix of the eye once or twice daily in the treatment of diabetic retinopathy. The implant softens and dissolves slowly, releasing the drug and absorbing it with the tear. Another example of such a device is supplied by the hydroxypropyl cellulose insert Lacrisert (Merck Inc.).

With targeted delivery systems, the active agent is isolated or concentrated in a localized area, tissue or site in the body for absorption or action.

The present invention also provides pharmaceutical dose delivery formulations and devices to increase the bioavailability of triethylenetetramine active agents. This may be accomplished in addition to or in combination with the dose releasing doses or devices described above.

Despite its good water solubility, triethylenetetramine is poorly absorbed from the digestive tract, and its bioavailability is incomplete and erratic, with great inter-individual variability. A therapeutically effective amount of the TET active agent is one capable of providing adequate TET levels in the blood. By increasing the bioavailability of the TET active, it is possible to reduce the dosage used to achieve therapeutic levels of the TET active.

Increased bioavailability of triethylenetetramine may be achieved by complexing or forming a bioavailability or absorption enhanced formulation of the triethylenetetramine active agent with one or more bioavailability or absorption enhancers .

The present invention provides additional agents for enhancing bioavailability or absorption for formulations of triethylenetetramine active agents. Such bioavailability or absorption enhancers include, but are not limited to, various surfactants such as various triglycerides such as cream, monoglycerides such as stearic acid and vegetable oils and esters, fatty acid esters, propylene glycol esters, polysorbate Alcohol Esters, Sodium Lauryl Sulfate, Sorbitan Esters, Dioctyl Sodium Sulfonate, and Other Substances. By modifying the surfactant properties of the delivery vehicle, the active agent can be exposed to the gastrointestinal tract for a longer period of time to increase absorption and reduce side effects. Further examples of such agents include carrier molecules such as cyclodextrins and their derivatives, whose well-known property is to act as complexing agents to alter the physicochemical properties of drug molecules. For example, cyclodextrins can stabilize (thermodynamically and oxidatively) or reduce the volatilization of the active agent, altering the solubility of the active agent after complexation. Cyclodextrin is a cyclic molecule comprising glucopyranose ring units and forming a curved torus structure. The cyclodextrin molecule is hydrophobic on the inside and hydrophilic on the outside, which makes the cyclodextrin molecule water-soluble. Solubility changes with substitution of hydroxyl groups on the exterior of the cyclodextrin molecule. Similarly, internal hydrophobicity can also change with substitution, although internal hydrophobicity is tuned within the cavity of the molecule. This adjustment between molecules is called complexation and produces inclusion complexes. Examples of cyclodextrin derivatives include thiobutyl cyclodextrin, maltocyclodextrin, hydroxypropyl cyclodextrin and salts thereof. The inclusion complexes of triethylenetetramine and carrier molecules such as cyclodextrin can reduce the dosage of triethylenetetramine to achieve therapeutic effect because of its ability to increase bioavailability.

The present invention provides microemulsions of triethylenetetramine active agent formulations with enhanced bioavailability. A microemulsion is a flowable, stable and homogeneous solution consisting of four major components, namely a hydrophilic phase, a lipophilic phase, at least one surfactant (SA) and at least one co-surfactant (CoSA). . A surfactant is a chemical substance that has two groups, one polar or ionic with a strong affinity for water, and the other with long or short aliphatic chains that are hydrophobic. These hydrophilic chemicals are capable of forming particles in aqueous or oily solutions. Suitable surfactants include mono-, di- and tri-glycerides, and polyethylene glycol (PEG) mono- and diesters. A co-surfactant, sometimes called a "co-surfactant," is a chemical substance with hydrophobic properties that facilitates the mutual dissolution of the water and oil phases of a microemulsion. Examples of suitable co-surfactants include ethyl diethylene glycol, propylene glycol lauryl, polyglyceryl oleate and related materials.

The present invention provides various polymers for triethylenetetramine active agent formulations to increase bioavailability by increasing the adhesion to mucosal surfaces, reducing the hydrolysis or enzymatic rate of the active agent, and increasing the surface area of the active agent particles. Suitable polymers may be natural or synthetic, and may be biodegradable or non-biodegradable. Delivery of low molecular weight active agents such as triethylenetetramine active agents can be accomplished by diffusion or degradation of the polymer system. Representative natural polymers include proteins such as zein, modified zein, casein, gelatin, gluten, serum albumin and collagen, polysaccharides such as cellulose, dextran and polyhyaluronic acid. Synthetic polymers are generally preferred for better degradation and release efficiency. Representative synthetic polymers include polyphosphazenes, polyvinyl alcohol, polyamides, polycarbonates, polyacrylates, polyolefins, polyacrylamides, polyenols, polyoxyalkylenes, polyalkylene p-phenylene Diacid salts, polyvinyl ethers, polyvinyl esters, polyvinyl halides, polyvinylpyrrolidone, polyglycolide, polysiloxane, polyurethane and their copolymers. Suitable polyacrylates include poly(methyl methacrylate), poly(ethyl methacrylate), poly(butyl methacrylate), poly(isobutyl methacrylate), poly(hexyl methacrylate), poly (isodecyl methacrylate), poly(lauryl methacrylate), poly(phenyl methacrylate), poly(methyl acrylate), poly(methyl isopropyl acrylate, and poly(octadecyl acrylates). Artificially modified natural polymers include cellulose derivatives such as alkyl cellulose, hydroxyalkyl cellulose, cellulose ethers, cellulose esters and nitrocellulose. Examples of suitable cellulose derivatives include formazan cellulose, ethyl cellulose, hydroxypropyl cellulose, hydroxypropyl methyl cellulose, hydroxybutyl methyl cellulose, cellulose acetate, cellulose propionate, cellulose acetate butyrate, cellulose acetate phthalate, Carboxymethylcellulose, cellulose triacetate, and sodium cellulose sulfate. Each of the polymers described above are commercially available from Sigma Chemical Co., St.Louis, Mo., Polysciences, Warrenton, Pa. , Aldrich Chemical Co., Milwaukee, Wis., Fluka, Ronkonkoma, N.Y., and BioRad, Richmond, Calif., or can be prepared synthetically using standard techniques from monomers obtained from the above suppliers. The polymers described above can be Divided into biodegradable, non-biodegradable and bioadhesive polymers, which are described in more detail below. Representative synthetic degradable polymers include polyhydroxy acids such as polylactic acid compounds, polyglycolic acid cross Esters and their copolymers, poly(ethylene terephthalate), poly(butic acid), poly(valeric acid), poly(lactide-cocaprolactone), polyanhydrides, polyorthoesters Substances and their mixtures and copolymers. Representative natural biodegradable polymers include polysaccharides such as alginate, dextran, cellulose, collagen and their chemical derivatives (replacement of chemical groups, addition of synthesis, such as alkylation, alkenylation, hydroxylation, oxidation and other modifications well known to those skilled in the art), proteins such as albumin, zein and their copolymers and mixtures, which can be used alone or with synthetic Polymer binding applications. Typically, these substances can be enzymatically hydrolyzed in vivo or exposed to water, causing their surface or backbone to erode. Examples of non-biodegradable polymers include vinyl acetate, poly(meth)acrylic acid, polyamide , polyethylene, polypropylene, polystyrene, polyvinyl chloride, polyvinyl phenol and their copolymers and blends. Hydrophilic polymers and hydrogels have bioadhesive properties. Containing carboxyl groups (such as poly [acrylic acid]) have the best bioadhesive properties. When soft tissue bioadhesion is required, polymers containing the highest concentration of carboxyl groups are preferred. Various cellulose derivatives such as alginic acid Sodium, carboxymethylcellulose, hydroxymethylcellulose, and methylcellulose also have good bioadhesive properties. Some of these bioadhesive substances are water-soluble and others are hydrogels. Polymers such as hydroxypropylmethylcellulose acetate succinate (HPMCAS), cellulose acetate-1,2,4-trimesate (CAT), cellulose acetate phthalate (CAP), hydroxypropylcellulose Acetate phthalate (HPCAP), hydroxypropylmethylcellulose acetate phthalate (HPMCAP) and methylcellulose acetate phthalate (MCAP) can be used in complexing Enhance the bioavailability of drugs. Rapidly bioerodible polymers such as poly(lactide-co-glycolide), polyanhydrides, and polyorthoesters, whose carboxyl groups are exposed on the outer surface, can also be used in bioadhesive drug delivery systems. In addition, polymers containing labile linkages, such as polyanhydrides and polyesters, are hydrolytically active, and their rate of hydrolytic degradation often varies with simple changes in the polymer chain. Upon degradation, these substances expose carboxyl groups on their outer surface and thus can be used in bioadhesive drug delivery systems.

Other agents that increase bioavailability or absorption can be used to facilitate or inhibit drug transport across the gastrointestinal mucosa. For example, blood flow in the gastrointestinal tract is a factor that determines drug absorption and bioavailability in the gastrointestinal tract, so agents that increase blood flow, such as vasodilators, can increase the absorption of orally administered drugs. Vasodilators can be combined with other drugs. For example, in EPO publication 106335 it is described that the coronary vasodilator diltiazem increases the bioavailability of drugs whose oral bioavailability does not exceed 20%, such as adrenergic beta-blockers (e.g. propranolol), catecholamines Substances (such as dopamine), benzodiazepine derivatives (such as diazepam), vasodilators (such as isosorbide dinitrate, nitroglycerin, or amylnitrosate), cardiotonic or antidiabetic agents, bronchodilators (such as Tetrahydroisoquinoline), hemostatics (such as chromium carbazole sulfonate), antispasmodics (such as timepidium halides) and antitussives (such as diphiperidine). Vasodilators thus constitute another class of agents that can enhance the bioavailability of triethylenetetramine.

Other mechanisms to increase the bioavailability of the compounds and formulations of the invention include reversed active transport inhibition mechanisms. For example, the active transport mechanism in intestinal epithelial cells is now thought to be the p-glycoprotein transport mechanism, which facilitates the reversal of material transport, allowing substances diffused or transported into the epithelial cells to return to the intestinal lumen. It can be speculated that p-glycoprotein on intestinal epithelial cells acts as a protective inversion pump, preventing absorption of toxic substances diffused or transported to the epithelial cells into the systemic circulation. A disadvantageous aspect of p-glycoprotein is that it can also prevent the utilization of useful substances, such as certain drugs, for which the p-glycoprotein antiporter system happens to work. Inhibition of p-glycoprotein causes less drug to return to the intestinal lumen and thus increases the amount of drug transported across the intestinal epithelium and can increase final plasma drug concentrations. Various p-glycoprotein inhibitors are well known in the art. These include water-soluble vitamin E; polyethylene glycols; poloxamers including Pluronic F-68; polyethylene oxides; polyoxyethylene castor oil derivatives including Cremophor EL and Cremophor RH 40; +)-Dihydroquercetin; naringenin; diosmin; quercetin, etc.

Inhibition of the reversal of the active transport system of active agents such as triethylenetetramine may increase the bioavailability of said active agents.

Surprisingly, as shown in Example 2 and Figures 3 and 4, triethylenetetramine dihydrochloride was able to effectively scavenge Cu in diabetic rats at a much lower dose than the prior art considered effective amount. Cu excretion normalized by body weight as can be seen in accompanying drawing 3, especially accompanying drawing 4, in the urine of diabetic rats after administration of triethylenetetramine 0.1 mg.kg ^-1 (in this study The Cu excretion of the lowest dose) was significantly more than that of diabetic rats administered with saline.

These data show that triethylenetetramine active agents including but not limited to triethylenetetramine, triethylenetetramine salts, triethylenetetramine analogs represented by ^general formulas I and II, etc. A dose of ¹ is effective in treating heart disease in humans. Effective doses can be applied in the order of 1/10, 1/100, 1/1000 (eg 120 mg.d ^-1 , 12 mg.d ^-1 , or 1.2 mg.d ^-1 ).

The present invention provides low dose delivery formulations and devices comprising one or more triethylenetetramine active agents, including but not limited to triethylenetetramine, triethylenetetramine salts, triethylenetetramines of general formulas I and II Ethylenetetramine analogs, etc., administered at an effective dose, for example, the dose range is 0.01mg.kg ^-1 -5mg.kg ^-1 , 0.01mg.kg ^-1 -4.5mg.kg ^-1 , 0.02mg.kg ^{- 1} -4mg.kg ^-1 , 0.02-3.5mg.kg ^-1 , 0.02-3mg.kg ^-1 , 0.05-2.5mg.kg ^-1 , 0.05-2mg.kg ^-1 , 0.05-0.1mg.kg ^-1 to 5mg.kg ^-1 , 0.05-0.1mg.kg ^-1 to 4mg.kg ^-1 , 0.05-0.1mg.kg ^-1 to 3mg.kg ^-1 , 0.05-0.1mg.kg ^-1 to 2mg.kg ^{- 1} , 0.05-0.1 mg.kg ⁻¹ to 1 mg.kg ⁻¹ , and/or any other value within this range.

Any such dosages for any route of administration or form of administration are described herein. It will be appreciated that any dosage delivery formulation or device described herein, particularly for oral administration, may be used in a delivery formulation or device for administration by any route, where available or desired. For example, the dosage forms of the present invention may be administered parenterally, or in specific oral dosage forms, such as sustained release dosage forms, extended release dosage forms, delayed release dosage forms, slow release dosage forms or repeated action oral dosage forms.

Another aspect of the invention is based on the results of studies on STZ rats excreting copper levels equivalent to humans, said dosage forms containing less than 250 mg of triethylenetetramine dihydrochloride (or triethylenetetramine dihydrochloride in the form of triethylenetetramine dihydrochloride The triethylenetetramine activator represented by compound). Capsules or tablets or any suitable dosage form of less than 250 mg of triethylenetetramine dihydrochloride or equivalent triethylenetetramine active agent may be used.

"Risk" as used herein refers to the assessment of the likelihood of disease in mammals, as described in JAMA, May 16, 2001, Volume 285 No. 19, 2486-2497, where the Framingham risk score Taking into account age, total cholesterol, HDL cholesterol, systolic blood pressure, treatment of hypertension, and whether or not smoking was also involved, plus any condition of glucose abnormality mentioned here.

As used herein, "elevation" refers to elevations in copper levels in mammals, such as humans, including undesired copper levels, removal of copper for therapeutic purposes, and/or at least about 10 mcg free copper/dL serum, applying Assay method of Merck & Co Inc.

Histological evidence of protection of the Wistar rat heart from diabetic injury (cardiomyopathy) after six months of treatment with TET, as judged by histology. The dose of triethylenetetramine is required to study urinary excretion of copper and iron, for example to study differences in excretion in diabetic and non-diabetic animals. For example, copper and iron excretion patterns were compared in normal and diabetic rats following acute intravenous administration of triethylenetetramine. In addition, the cardiovascular side effects of acute intravenous administration of triethylenetetramine can be determined.

For a better understanding of the present invention, the following experimental part will be introduced. The following experiments are only used to illustrate the present invention, and do not limit the present invention in any way.

Example 1

This example was used to determine baseline physiological data for streptozotocin (STZ) treated rats compared to baseline physiological data for diabetic and non-diabetic rats.

All methods used in this study were approved by the University of Auckland Animal Ethics Committee and complied with The Animals Protection Act and Regulations of New Zealand.

To induce diabetes, male Wistar rats (n=28, 303±2.9 g) were randomly divided into diabetic and non-diabetic groups. After anesthesia (5% trifluorobromochloroethane and 2l.min ^-1 of O ₂ ), animals in the diabetic group received a single dose of streptozotocin (STZ, 55 mg.kg ^-1 (body weight), via the tail vein. Sigma; St. Louis, MO) in 0.5 ml saline solution. Non-diabetic animals received an equal volume of saline. After injection, both groups of rats were housed under similar conditions and provided with normal rat chow (Chow 86 pellets; New Zealand dry food, Auckland, NZ) and deionized water. Blood glucose and body weight were measured three days after STZ/saline injection in the study and then weekly thereafter. Diabetic rats showed polydipsia, polyuria and hyperglycemia (>11 mmol.l ^-1 , Advantage II, Roche Diagnostics, NZ Ltd).

The result is shown below. Considering the effect of STZ on blood glucose and body weight, blood glucose increased to 25±2mmol.l ^- 1 three days after STZ injection (Table 1). Diabetic animals lost more weight than the non-diabetic group over the 44 consecutive days following STZ/saline injection, even though they ate more food per day. On the day of the experiment, the blood glucose levels of diabetic and non-diabetic animals were 24±1 and 5±0mmol.l ^-1 and the body weights were 264±7g and 434±9g, respectively.

Table 1. Blood Glucose, Body Weight and Food Consumption in Diabetic vs Non-diabetic Animals diabetes group non-diabetic group Body weight before STZ/saline injection 303±3g 303±3g Blood glucose three days after STZ/saline injection ^* 25±2mmol.l ^-1 5±0.2mmol.l ^-1 daily food consumption ^* 58±1g 28±1g Blood glucose on the experiment day ^* 24±1mmol.l ^-1 5±0.2mmol.l ^-1 Body weight on the experiment day ^* 2647g 434±9g

Diabetic animals n=14, non-diabetic animals n=14. Listed values are ± SEM mean. ^* indicates significant difference (P<0.05).

Thus, the results showed that STZ treatment caused an increase in blood glucose, an increase in food intake, and a decrease in body weight in the diabetic group.

Example 2

This example evaluates urinary excretion of copper and iron in diabetic and non-diabetic groups of rats acutely administered increasing doses of triethylenetetramine intravenously.

Six to seven weeks (average 44±1 days) after STZ administration, animals were recorded for control or drug experiments. All animals were fasted overnight before surgery, but were fed deionized water ad libitum. Anesthesia was administered and maintained with 3-5% trifluorobromochloroethane and 2 l.min ^-1 of O ₂ . The femoral artery and vein were cannulated with a solid blood pressure transducer (MikrotipTM 1.4F, Millar Instruments, Texas, USA) and a saline-filled PE 50 catheter. The ureter was exposed by midline laparotomy, cannulated with a polyethylene catheter (0.9 mm external diameter, 0.5 mm internal diameter), and the wound was closed. Catheterized animals were ventilated with 70-80 breaths per minute and supplemented with _O2 (pressure-controlled ventilator, Kent Scientific, Connecticut, USA). Respiration rate and intratidal pressure (10-15 _cmH2O ) were adjusted to maintain intratidal _CO2 pressure at 35-40 mmHg (SC-300 _CO2 monitor, Pryon Corporation, Wisconsin, USA). During the operation and experiment, the body temperature was maintained at 37°C with a heating plate. Estimated fluid losses were supplemented intravenously with 154mmol.l ^-1 NaCl solution at a rate of 5ml.kg ^-1 .h ^-1 .

Stabilize for 20 min after surgery before starting the experiment. Triethylenetetramine was administered intravenously over 60 seconds at increasing concentrations (0.1, 1.0, 10 and 100 mg.kg ^-1 in 75 μl of saline with 125 μl of saline flush). The control group received an equal volume of saline. Urine was collected during the experiment at 15 min intervals in pre-weighed polyethylene epindorf tubes. Terminal blood samples were collected by cardiac puncture at the end of the experiment, and serum was separated and stored at -80°C until analysis. Isolate the heart with a rapid midline sternotomy and proceed with the following procedure.

Mean arterial pressure (MAP), heart rate (HR, available from the MAP waveform), oxygen saturation (Nonin 8600V pulse oximeter, Nonin Medical Inc., Minnesota, USA) and core body temperature were monitored continuously throughout the experiment. Monitoring was performed using a PowerLab/16s data acquisition module (AD Instruments, Australia). The scaled signal is displayed on the screen and saved as a 2s average for each variable.

Urinalysis and tissue analysis were performed as follows. Instrument: Perkin Elmer (PE) Model 3100 atomic absorption spectrophotometer equipped with PE HGA-600 graphite furnace and PEAS-60 automatic sampler for the determination of Cu and Fe in urine. Apply deuterium background correction. Cu or Fe hollow cathode lamps (Perkin Elmer Corporation) were used and operated at 10W (Cu) or 15W (Fe). The atomic line at 324.8 nm is for Cu and the atomic line at 248.3 nm is for Fe. The slit widths of both Cu and Fe are 0.7 nm. Pyrolytically coated graphite tubes were used for all analyses. The injection volume was 20 μl. Typical graphite furnace temperature conditions are as follows:

GF-AAS temperature program process temperature °C RamP/s Hold/s Internal flow rate/ml.min ^-1 Drying Pretreatment Atomization - Cu/Fe Posttreatment 901201250 ^* 202300/25002600 16020111 55101085 3003003003000300

^* The pretreatment temperature is 1050°C for the analysis of tissue digestion (see Example 3)

Reagents : All reagents were of the highest purity, at least analytical grade. The standard working solution of Cu and Fe of GF-AAS was diluted stepwise to obtain 1000 mg.l ^-1 (Spectrosol standard solution; BDH). Water purification was accomplished with a Millipore Milli-Q ultra-purity water system with a resistivity value of 18 MΩ measured.

Urine sample pretreatment : Urine was collected in pre-weighed 1.5 ml microtubes (eppendorf). After reweighing, the urine samples were centrifuged and the supernatant was diluted 25:1 with 0.02M 69% Aristar grade _HNO3 . Samples were stored at 4°C prior to GF-AAS analysis. If samples need to be stored for longer than two weeks, lyophilize and keep at -20°C. Serum : Terminal blood samples were centrifuged and serum was stored as urine until analysis. Serum trace metals in blood samples were collected at the end of the experiment, and renal clearance was calculated using the following equation:

Do the following statistical analysis. All values are expressed as ±SEM mean and P-value <0.05 indicates statistical significance. Unpaired t-tests were initially used to detect differences in weight and glucose between diabetic and control groups. To compare drug responses, statistical analysis was performed using analysis of variance (statistical values, see Windows v.6.1, SAS Institute Inc., Calfornia, USA). Subsequent statistical analysis was performed using a mixed model with repeated detection analysis of variance ANOVA (see Example 4).

The result is as follows. With regard to cardiovascular changes during infusion, baseline levels of MAP in pre-infusion controls were not significantly different between diabetic and non-diabetic animals (99 ± 4 mmHg). HR in the diabetic group was significantly lower than that in the non-diabetic group (287±11 and 364±9 bpm, respectively, P<0.001). Infusion of triethylenetetramine or saline had no effect on these variables, except at the maximum dose, where MAP decreased to 19 ± 4 mmHg within 2 minutes post-dose and returned to pre-dose levels within 10 minutes. Body temperature and oxygen saturation were kept stable during the experiment.

Regarding urinary excretion, except at the maximum drug dose (100 mg.kg ^-1 ) or an equal volume of saline (Fig. 16), diabetic animals excreted significantly more urine than non-diabetic groups. Compared with the non-diabetic group given the same volume of saline, the administration of 100 mg.kg ^-1 triethylenetetramine can also increase the urine excretion of animals in the non-diabetic group (Fig. 17). This effect was not found in the diabetic animal group.

Analysis of the urinary excretion response drug curves for Cu and Fe showed that at all doses, diabetic and non-diabetic animals receiving drug excreted more Cu than the group receiving an equal volume of saline alone (Figure 18). To correct for the weight effect of diabetic animals, and thus allow more comparisons between diabetic and non-diabetic animals, excretion rates of trace elements were also calculated per gram of body weight. Figure 19 shows that the copper excretion of diabetic animals was significantly higher than that of non-diabetic animals after receiving the drug. The same thing happened in the comparison with the group that received saline, but the effect was not very significant.

Excretion of total copper was significantly increased in the diabetic and non-diabetic groups of animals receiving triethylenetetramine compared to the control group receiving saline over the entire period of the experiment (Fig. 20). Diabetic animals that received the drug also excreted more copper per gram of body weight than the non-diabetic group. The same significant effect was also observed in the group of animals given saline (Fig. 21).

In comparison, iron excretion was not increased in diabetic and non-diabetic animals receiving triethylenetetramine compared to animals receiving an equal volume of saline (Fig. 22). Analysis per gram of body weight showed that diabetic animals receiving saline excreted significantly more iron than non-diabetic animals, but this was not evident in both diabetic and non-diabetic groups receiving triethylenetetramine (Fig. 23). Excretion of total iron in diabetic and non-diabetic animals receiving drug did not differ from the group receiving saline (Fig. 24). Consistent with the analysis of dose-response curves, iron excretion per gram of body weight was significantly higher in diabetic animals receiving saline than in non-diabetic animals, but this was not observed in the triethylenetetramine group (Fig. 25).

Electron paramagnetic resonance spectroscopy showed that urinary Cu of drug-treated animals mainly formed triethylenetetramine-Cu ^II (FIG. 28), indicating that the increased Cu in tissues of diabetic rats was mainly divalent. These data indicate that Cu ^II is increased in severely hyperglycemic rats and can be excreted via selective chelation.

Regarding serum content and renal clearance of Cu and Fe, where there was no significant difference in serum copper content, there was a significant increase in renal clearance in diabetic animals receiving drug compared to diabetic animals receiving saline (Table 2) . The same was true for non-diabetic animals, but the trend was not statistically significant (P=0.056). There was no effect on iron serum content or renal clearance.

Table 2. In diabetic and non-diabetic animals receiving drug or saline

Serum content and renal clearance of Cu and Fe 1.1.aa1 Diabetes 1.1.aa1 Diabetes 1.1.aa2 Non-diabetic 1.1.aa2 Non-diabetic Triethylenetetramine n=6 brine n=7 Triethylenetetramine n=4 brine n=7 Serum Cu (μg.μl ^-1 ×10 ^-4 ) 7.56±0.06 9.07±1.74 7.11±0.41 7.56±0.62 Serum Fe (μg.μl ^-1 ×10 ^-4 ) 35.7±7.98 63.2±16.4 33.6±1.62 31.4±8.17 Cu renal clearance (μl.min ^-1 ) ^* 28.5±4.8 1.66±0.82 19.9±6.4 0.58±0.28 Fe renal clearance rate (μl.min ^-1 ) 0.25±0.07 0.38±0.15 0.46±0.22 0.11±0.03

Data are mean ± SEM. ^* Asterisks indicate significant differences (P<0.05) between diabetic animals receiving triethylenetetramine and diabetic animals receiving an equal volume of saline.

Briefly, acute intravenous administration of triethylenetetramine significantly increased total copper excretion in both diabetic and nondiabetic groups compared with saline controls. Furthermore, following acute intravenous administration of increasing doses of triethylenetetramine, copper excretion per gram of body weight was significantly increased in diabetic animals compared with non-diabetic animals. In contrast, total iron excretion was not significantly different between diabetic animals that received the drug and non-diabetic animals that received saline.

Example 3

This example was used to determine the effect of acute intravenous administration of increasing doses of triethylenetetramine on the copper and iron content of normal and diabetic cardiac tissue.

Methods as below. Spectrophotometric analysis was as described in Example 2. Cu, Fe and Zn in digested tissues were determined at Hill Laboratories (Hamilton, New Zealand) using PE Sciex Elan-6000 and PE Sciex Elan-6100 DRC ICP-MS. The operating parameters are listed in the table below.

Instrument Operating Parameters for ICP-MS parameter value Inductively Coupled Plasma RF Energy Argon Plasma Gas Flow Rate Argon Auxiliary Gas Flow Rate Argon Spray Gas Flow Rate Interface Sample Cone and Orifice Diameter Separator Cone and Orifice Diameter Data Acquisition Parameters Scan Mode Sampling Time Scan/Repeat Sample Absorption Rate 1500W15l.min ^-1 1.2l.min ^-1 0.89l.min ^-1 Ni/1.1mmNi/0.9mm peak jump 30ms(Cu, Zn)/100ms(Fe)2031ml.min ^-1

Reagents are as follows. Standard Reference Material 1577b Bovine liver was obtained from the National Institute of Standards and Technology and used to evaluate the effect of tissue digestion. The results obtained are as follows:

GF-AAS and ICP-MS results of NIST SRM 1577b bovine liver* element determined value GF-AAS ICP-MS CuFeZn 160±8184±15127±16 142±12182±21- 164±12166±14155±42

^* Indicates that the unit of measurement for dry matter is μg.g ^-1 .

Sample preprocessing is as follows. Heart : After isolation, wash to remove excess tissue, remove excess blood with buffer solution, blot dry, and record the wet weight of the ventricle. The left ventricular muscle was dissected using a titanium tool and placed in a pre-weighed 5.0 ml polystyrene tube. The sample was freeze-dried overnight to constant weight, and 0.45 ml of analytical grade 69% HNO ₃ was added. The sample tubes were heated in a water bath at 65°C for 60 minutes. Take 4.5ml of sample and add milli-Q H ₂ O. The resulting solution was diluted 2:1 to reduce the concentration of _HNO3 below the maximum allowed by ICP-MS analysis.

The result is as follows. With regard to the metal content of cardiac tissue, the wet weight of the heart was significantly smaller in diabetic animals than in non-diabetic animals, while the heart/body weight ratio was increased (see Table 3). The Cu and Fe contents of heart tissue from some animals were also analyzed. There were no significant differences in copper content between the diabetic and non-diabetic groups that received saline or triethylenetetramine. Iron content was significantly higher in the non-diabetic group receiving saline than in the diabetic group receiving saline (see Table 3).

Table 3: Heart weight, heart/body weight ratio and

Comparison of Trace Metal Contents in Heart Tissue diabetes group non-diabetic group heart wet weight ^* 0.78±0.02g 1.00±0.02g heart/body weight ratio ^* 2.93±0.05mg.g ^-1 2.30±0.03mg.g ^-1 Cu content in dried tissues μg.g ^-1 Triethylenetetramine treatment Saline treatment 24.7±1.521.3±0.9 27.1±1.027.2±0.7 Fe content in dried tissues μg.g ^-1 Triethylenetetramine treatment Saline treatment 186±46180±35 235±39274±30

Diabetic animals: n=14; non-diabetic animals: n=14. Values are ± SEM mean. ^* Asterisks indicate significant differences (P<0.05) between diabetic and non-diabetic animals.  indicates a significant difference (P<0.05) between diabetic and non-diabetic animals receiving saline.

Briefly, it was demonstrated that acute intravenous administration of increasing amounts of triethylenetetramine had no significant effect on the copper content of diabetic and normal cardiac tissue.

Example 4

In this example, a mixed linear model was performed on the data in Examples 1-3 above.

Methods as below. Regarding statistical analysis using mixed linear models, the data for each dose level were analyzed using mixed linear models (PROC MIXED; SAS, Version 8). The model included diabetes, drug, and their interaction as fixed effects, time as repeated measures, and rats as subjects of the dataset. Objects are assumed to be completely self-contained. Complete models were fitted to each dataset applying maximum likelihood estimation (REML) to mixed linear models (i.e. fixed and random effects modulo). Mixed models are a general generalization of standard linear models and can be used to analyze data on a wide variety of variables. The significance level in the whole test is 0.05. The result is as follows.

For copper, diabetic rats excreted significantly higher levels of copper at all dose levels (see Figure 27). Baseline copper excretion was also significantly higher in diabetic rats than in non-diabetic rats. There were no differences between the drug and saline groups at baseline. Significant interactions in the model were at dose levels 1.0 mg.kg ^-1 and higher. The presence of significance indicates that changes in one effect are influenced by the other. Thus, a significant interaction between diabetes and drug factors resulted in increased copper excretion over the predicted effects of both factors.

For iron, diabetic rats receiving saline excreted significantly higher iron than the other dose levels. This resulted in significant increases in all factors in the model at all dose levels.

Briefly, the acute effects of intravenous triethylenetetramine on the cardiovascular system and the urinary excretion of copper and iron were investigated in anesthetized diabetic (6-week streptozotocin-induced diabetic) and non-diabetic rats. studied. Animals were divided into four groups: diabetic + TET, diabetic + saline, non-diabetic + TET, non-diabetic + saline. Drugs or an equal volume of saline were administered hourly in doses of increasing strength (0.1, 1.0, 10, 100 mg.kg ^-1 ), and urine was collected every 15 minutes during the experiment. Finally blood samples were taken and heart tissue was obtained. Urine analysis showed that: (1) at all doses, diabetic and non-diabetic animals receiving drug excreted more Cu (μmol) than animals receiving an equal volume of saline; (2) for analysis per gram of body weight, each three The excretion of copper (μmol.gBW ^-1 ) in diabetic animals was significantly higher than that in non-diabetic animals. The same situation was also observed in the saline group, but it was not significant at every dose; (3) For most doses, iron excretion (μmol) was more in the diabetic animal group receiving saline than in the drug group. In the excretion of iron in the non-diabetic group, there was no difference between the saline group or the triethylenetetramine group; (4) analysis per gram of body weight showed no difference in iron excretion between diabetic and non-diabetic animals receiving triethylenetetramine . Diabetic animals that received saline excreted more iron per gram of body weight than non-diabetics; (5) cardiac tissue analysis showed no significant difference in total copper content between diabetic and non-diabetic animals, and drug effects on cardiac iron and copper content There was also no effect; and (6) Calculations of renal clearance showed significantly higher copper clearance in diabetic animals receiving triethylenetetramine than in diabetic animals receiving saline. The same situation was also present in non-diabetic animals, but to a lesser extent. Triethylenetetramine had no effect on iron scavenging.

No adverse cardiovascular effects were observed after acute administration of triethylenetetramine. Triethylenetetramine treatment effectively increased copper excretion in both diabetic and non-diabetic animals. Urinary copper excretion per gram of body weight after administration of triethylenetetramine was greater in diabetic animals than in non-diabetic animals. Triethylenetetramine does not increase iron excretion in diabetic or nondiabetic animals.

Example 5

An experiment was performed on the effect of triethylenetetramine on restoring cardiac function in STZ diabetic rats. As mentioned above, there is histological evidence that treatment with triethylenetetramine can protect the hearts of diabetic Wistar rats with cardiac injury (diabetic cardiomyopathy). However, it is unclear whether improvements in histology lead to improvements in cardiac function.

This experiment is to compare the cardiac function of STZ diabetic rats treated or untreated with triethylenetetramine and normal rats, using an isolated heart model.

Methods as below. Experimental animals were cared for according to the "Code of Care of Laboratory Animals" (National Association for Medical Research) and the University of Auckland Animal Ethics Committee approved the study.

Male albino Wistar rats weighing 330-430 g were divided into four groups, as shown in Table 4.

Table 4. Experimental grouping Group code name N treat Group A STZ 8 13 weeks of diabetes Group B STZ/D7 8 13 weeks of diabetes (medication for 7-13 weeks) Group C Sham 9 non-diabetic control Group D Sham/D7 11 Non-diabetic control (drug treatment for 7-13 weeks)

STZ = streptozotocin; D7 = 7 consecutive weeks of triethylenetetramine treatment, starting 6 weeks after the start of the experiment.

Diabetes was induced by intravenous streptozotocin (STZ; Sigma; St. Louis, MO). All rats were subjected to brief inhalational anesthesia (inhalation: 5% bromotrifluoroethane and 2 L/min oxygen, and maintenance 2% trifluorobromochloroethane and 2 L/min oxygen). Two diabetic groups received a single intravenous bolus injection of STZ (57 mg/kg body weight) in 0.5 ml of 0.9% saline solution via the tail vein. Non-diabetic placebo-treated animals received an equal volume of 0.9% saline solution. Diabetic and non-diabetic rats were housed in pairs and had ad libitum access to a normal diet (chow 86 pellets; New Zealand dry food, Auckland, NZ) and deionized water ad libitum. There are two water bottles per cage, giving rats an equal opportunity to drink water or drugs. The ambient temperature of the animals is 21°C, the humidity is 60%, and the floor of the standard mouse cage is covered with sawdust, which is replaced daily.

Blood samples were drawn from tail tip capillaries for determination of blood glucose (Advantage II, Roche Diagnostics, NZ Ltd). All groups were sampled simultaneously. Blood glucose and body weight were measured on day 3 after STZ/saline injection and then weekly throughout the study. Diabetes was determined by the presence of polydipsia, polyuria and hyperglycemia (>11 mmol.L ^-1 ).

In the drug-treated diabetes group, triethylenetetramine was given to each cage at a concentration of 50 mg/L in drinking water. Drinking water containing triethylenetetramine was given continuously from the seventh week until the animals were sacrificed at the end of the thirteenth week. For the Sham/D7 non-diabetic group who drank less water than the diabetic group, the drug concentration in their drinking water was adjusted accordingly, so that they consumed approximately the same dose of drug as the STZ/D7 group. The average daily intake of TET-treated animals was 8-11 mg.

When drug was initiated in the diabetic group, the diabetic animals were expected to have cardiomyopathy, as predicted in the original study (data not shown) and confirmed in the known literature. See Rodrigues B, et al., Diabetes 37(10):1358-64 (1988).

On the last day of the experiment, animals were anesthetized (5% bromochlorotrifluoroethane and 2 L/min of oxygen) and heparin (500 IU.kg ^-1 ) (Weddel Pharmaceutical Ltd., London) was administered via tail vein. A 2 ml blood sample was collected from the inferior vena cava, and the heart was rapidly isolated and immersed in Krebs-Henseleit bicarbonate buffer on ice to preserve its contractile activity. The heart is then placed in an isolated cardiac perfusion device.

The aortic root of the heart is quickly ligated to the aortic cannula of the perfusion set. Retrograde (Langendorff) perfusion was performed at a static pressure of 100 cm H ₂ O and a temperature of 37° C. for 5 minutes, and cannulation of the left atrium via the pulmonary vein was completed. The non-working (Langendorff) preparation was then converted to a working heart model by flowing perfusion buffer from the aorta to the left atrium at a filling pressure of ₁₀ cmH2O. The left ventricle ejects automatically towards the aortic cannula at a static pressure (afterload) of 76 cm _H2O (55.9 mmHg). The perfusate was K-Hahn's bicarbonate buffer (mM: KCl 4.7, CaCl ₂ 2.3, KH ₂ PO ₄ 1.2, MgSO ₄ 1.2, NaCl 118, and NaHCO ₃ 25), pH 7.4, containing 11 mM Glucose, and continuously filled with 95% O ₂ : 5% CO ₂ . Buffers can also be filtered in a streamline (8 [mu]m at first followed by 0.4 [mu]m cellulose acetate filtration; Sartorius, Germany). The temperature of the perfusion apparatus was maintained with a water jacket, and the temperature of the buffer was continuously monitored and maintained at 37 °C during the perfusion process.

A modified 24 g plastic venous cannula (Becton Dickson, Utah, USA) was inserted through the apex of the heart into the left ventricle with a conventional insertion needle. The cannula was then attached to a SP844 piezo pressure transducer (AD device) for continuous monitoring of left ventricular pressure. Aortic pressure was continuously monitored by pressure transducers on the side branches of the cannulated aorta (Statham Model P23XL, Gould Inc., CA, USA). The heart beat (Digitimer Ltd, Heredfordshire, England) at a frequency of 300 bpm was formed using suprathreshold voltage with the help of electrodes attached to the aorta and pulmonary vein cannula, and the pulse was 5-ms generated by a square wave.

Aortic flow was recorded by a linear flowmeter (Transonic T206, Ithaca, NY, USA) and coronary flow was measured by time-point outflow recorded by coronary vein at 30 s collection.

The various cardiac working devices used have been described in many ways, eg, Neely, JR, et al., Am J Physiol 212:804-14 (1967). The improved device can measure cardiac function under different preload pressures (Fig. 14 and Fig. 15). The construction of the corresponding device is completed, and the height of the buffer flow to the heart can be changed by a series of scales in a reconfigurable manner. For preload, the efferent canal of the aorta can also be increased in height to provide various defined afterload pressures . Afterload heights are defined in mm Hg, consistent with publications.

All data from pressure sensors and flow probes were collected (Powerlab16s data acquisition machine; AD Instruments, Australia). The data processing function of the device is used to calculate the first derivatives of the two pressure waves (ventricular and aortic). Final cardiac function data included:

Cardiac output ^* ; aortic blood flow; coronary blood flow; left ventricular apex/aortic pressure; maximal ratio of ventricular pressure (+dP/dt) ^** ; maximal ratio of ventricular diastolic pressure (-dP/dt) ^** ; maximum ratio of aortic pressure (aorta + dP/dt); maximum ratio of aortic diastolic pressure (aorta - dP/dt). [ ^* Cardiac output (CO) is the amount of buffer pumped by the heart per unit time, including the amount pumped by the aorta and the coronary vessels. This is a sign of full heart function. ^** +dP/dt is the ventricular rate of change (or aortic pressure) and is related to ventricular contractility (contractility). It can be used to compare the contractility of different hearts under the same preload (Textbook of Medical Physiology, Ed. A. Guyton. Saunders company 1986). -dP/dt is a recognized measure of ventricular diastolic rate].

The experiment is divided into two parts, the first part is to fix the afterload and change the preload, and the second part is to quickly follow the first part, and fix the preload and change the afterload.

Fixed afterload and variable preload : After completion of cannulation, the heart was equilibrated for 6 minutes under an atrial filling pressure of 10 cm H ₂ O and an afterload of 76 cm H ₂ O. During this period, a left ventricular pressure sensor cannula was inserted and the pulse started. Once the heart is stabilized, the atrial filling pressure is reduced to 5 cmH ₂ O and then gradually increased in 2.5 cm H ₂ O steps, reaching a maximum of 20 cm H ₂ O after 7 steps. Preload was maintained for 2 minutes at each filling pressure, during which time the pressure was kept stable and coronary blood flow was measured. After the preload change test is completed, the afterload change test part should be started quickly.

Fixed preload and variable afterload : In this part of the experiment, the filling pressure (preload) was set at 10 cm H ₂ O, and the afterload was increased from 76 cm H ₂ O (55.9 mm Hg), and each increase lasted 2 minutes , a total of 9 times. The maximum value (afterload) that each individual heart can bear depends on the maximum afterload value of 145cm H ₂ O (106.66mm Hg), or the measured value when the aortic blood flow becomes 0ml/min. In the latter case, the heart is considered "failed". To determine that the failure was indeed functional and not due to other causes (such as chronic ischemia or valve damage), all hearts were returned to initial perfusion conditions (preload 10 cm H ₂ O; afterload 75 cm H ₂ O) for 4 minutes to be sure of the restoration of functionality. At the end of the procedure, the heart was arrested by backperfusion with 4 ml of cold KCl (24 mM). The atrial and vascular remnants were removed, and the hearts were blotted dry and weighed. The ventricle is cut along the midline of the apex of the heart and the atrioventricular groove. Ventricular wall thickness was measured using a micro caliper (Absolute Digimatic, Mitutoyo Corp, Japan).

Data from the Powerlab were extracted by averaging 1 min intervals from the electron scan stabilization section produced at each step in the recording. Results from each group were combined and analyzed for differences between cardiac function parameters (aortic blood flow, cardiac blood flow, MLVDP, LV or aortic +/- dP/dt). Repeated observations for differences in preload conditions were explored using approximate repeated measures mixed models and compared across study groups (SAS v8.1, SAS Institute Inc, Cary NC). Applying maximum likelihood results in loss of randomness in the data. Significant mean and interaction effects were further validated using Tukey's method and maintaining a 5% error rate for pairs. All assays are two-tailed. Residue analysis was done using Proc Liftest (SAS V8.2). One-way ANOVA was used to detect differences in weight change parameters between groups. Tukey's test was used to compare the differences among the groups. Unless otherwise stated, ^* in each graph indicates p<0.05=STZ v STZ/D7, #.p<0.05=STZ/D7 v Sham/D7.

The results showing the weight of the animals at the end of the experiment are shown in Table 5. Diabetic animals were about 50% smaller than corresponding normal animals. The percentage change graph of the weight of each experimental group is shown in Figure 5, where the arrow indicates the beginning of the treatment with triethylenetetramine.

Table 5. Initial and final animal body weights (mean ± SD)

^* P＜0.05

The blood glucose values of the three groups of rats are shown in Figure 6. Generally, diabetes is established and confirmed 3-5 days after STZ injection. The placebo Sham and Sham/D7 control groups maintained normoglycemia during the experiment. There was no difference in blood glucose profile after treatment with the drug (p=ns) comparing the treated group with the non-treated control group.

The final heart weight and ventricular wall thickness measurements are shown in Table 6. Animals in the diabetes treated group had a small but significant improvement in the "heart: body weight" ratio. The "ventricular wall thickness: body weight" ratio tended to improve in the diabetic treatment group compared to the non-diabetic treatment group, but the effect did not reach significance.

Fix afterload and vary frontload

Figures 7-12 are graphs showing cardiac performance parameters (STZ Diabetes; STZ Diabetes+Drug; and placebo-treated controls) at increased atrial filling pressures (5-20 cmH ₂ O, preload) and constant afterload. In the case of a load of 75cm H ₂ O. All results are means ± SEM. Unless otherwise stated, only other groups with significant differences related to STZ/D7 are marked in each graph; ^* indicates p<0.05 for STZ v STZ/D7; #p<0.05 for STZ/D7 v Sham/D7. Unless otherwise stated, STZ/D7 v Sham or Sham/D7 were not significant.

Cardiac output (Fig. 7) is the sum of aortic blood flow (Fig. 10) and coronary blood flow (Fig. 8). Since the hearts of the control group and the experimental group have significant differences in final weight, the coronary blood flow is also the blood flow normalized to the weight of the heart (Fig. , and relative to the heart weight ratio).

Table 6. Final heart weight (g) and animal body weight (BW) per gram (mean ±)

^* P＜0.05

§＝Significantly different from STZ STZ/D7 group p<0.05

The first derivative of the pressure curve gives the rate of change of ventricular pressure during each cardiac cycle, and the maximum positive rate of change value (+dP/dt) is plotted, see Figure 11. The corresponding maximum relaxation rate (-dP/dt) is shown in Figure 12. Similar results showing improved cardiac function were obtained from data derived from aortic pressure catheterization (results not shown).

Fix front load and vary load

Under conditions of increasing afterload with constant preload, the ability of the heart to cope with increased afterload can be assessed. The curve of functional residual, that is, the amount remaining in the heart with aortic blood flow after each afterload, is shown in Fig. 13 .

Administration of triethylenetetramine improved cardiac function in STZ diabetic rats compared with untreated controls. For example, cardiac output, ventricular systole and diastole, and coronary blood flow were all improved in TET-treated diabetic rats compared to treated controls.

Example 6

This example was used to further evaluate the effect of acute administration of triethylenetetramine on cardiac tissue by assessing left ventricular (LV) histology.

Methods as below. Following functional analysis, histological aspects of LVs were investigated by laser focusing (LCM; Figures 29a-d) and transmission electron microscopy (TEM; Figures 29e-h). For LCM, LV sections were co-stained with muscarinin to visualize actin, and β-integrin, as a marker of the extracellular space. See Ding B, et al., "Left ventricular hypertrophyin ascending aortic stenosis in mice: anoikis and the progression to early failure," Circulation 101:2854-2862 (2000).

For each treatment, 5 sections from each of 3 hearts were examined with LCM and TEM. For LCM, LV sections were fixed (4% paraformaldehyde, 24 hours); implanted (6% agar); vibrating sectioned (120pm, Campden); filamentous actin stained (Phalloidin-488, Molecular Probes) and _β1 - Staining with integrin antibody and goat anti-rabbit secondary antibody conjugated to CY% (1:200; Ding B, et al. "Left ventricular hypertrophy inascending aortic stenosis in mice: anoikis and the progression to early failure," Circulation 101:2854-2862 (2000)); and Visualization (TCS-SP2, Leica). For TEM, samples were post-fixed (1:1 v/v 1% w/v 0s0 M 0s0 M PBS); stained (aqueous uranyl acetate (2% w/v, 20mm) followed by citrate (3mm) ); sectioning (70 nm); and imaging (CM-12, Phillips).

The result is as follows. Copper chelator normalizes LV in diabetic rats. Compared with controls (Fig. 29a), diabetes caused marked myocardial structural changes, marked by loss of cardiomyocytes; thinning and destruction of remaining myofibrils; decrease in actin filament density; and enlargement of interstitial spaces (attached Figure 29b). These findings have been reported. See Jackson CV, et al., "Afunctional and ultrastructural analysis of experimental diabetic ratmyocardium: manifestation of cardiomyopathy," Diabetes 34:876-883 (1985). Compared with the markers, the myocardial tissue after the treatment with triethylenetetramine was improved (Fig. 29c). Importantly, the orientation and volume of cardiomyocytes, as well as their actin, are largely normalized, consistent with -dP _LV /dt observed in functional studies. Triethylenetetramine treatment reverses enlargement of the cardiac ECM. Triethylenetetramine-treated non-diabetic myocardium was normal in LCM (Fig. 29d), indicating no observable adverse effects on LV structure. Therefore, Cu chelation can basically restore the normal tissue structure of the myocardium, but it does not eliminate hyperglycemia. These data provide structural relevance for the recovery of cardiac function as described above.

TEM largely agrees with LCM. Compared with controls (Fig. 29e), diabetes induced myocardial injury, characterized by loss of cardiomyocytes, with obvious myofiber collapse; destruction of remaining cardiomyocytes, manifested by mitochondrial enlargement, very obvious; Expand (Supplementary Figure 29f). These findings have been reported. See Jackson CV, et al., "Afunctional and ultrastructural analysis of experimental diabetic ratmyocardium: manifestation of cardiomyopathy," Diabetes 34:876-883 (1985). Oral administration of triethylenetetramine caused substantial restoration of diabetic LV structure with massive increase and normalized myocyte orientation; restoration of normal mitochondrial structure; and narrowing of the marked extracellular space (Fig. 29g) . These data suggest that hyperglycemia-induced accumulation of Cu ^II leads to mitochondrial dysfunction. See Brownlee M, "Biochemistry and molecular cell biology of diabetic complications," Nature 414:813-820 (2001). TEMs of TET-treated and non-diabetic myocardium were normal (Fig. 29h). Thus, TET treatment normalized hyperglycemia-induced myocardial injury at both the cellular and intercellular levels. Taken together, these microscopic studies provide remarkable evidence that selective Cu chelation can substantially improve LV structure, even in severe chronic hyperglycemia.

In brief, it was demonstrated that (1) treatment with TET had no significant effect (as expected) on blood glucose concentrations in the two diabetic groups; There was a small but significant improvement in the (heart weight)/(body weight) ratio in the treated diabetic group; (3) Cardiac output was restored for Sham values with increased foreload and unchanged afterload. Both aortic and coronary blood flow were improved in the drug treatment group; (4) Ventricular systole and diastole were significantly improved in the drug treatment group. This improvement restored function to some extent, but there was no significant difference between the drug-treated group and the Sham control group; (5) Changes in the pressure measured by the aortic sensor also showed that the drug-treated diabetic group was comparable to the untreated diabetic group. Improvement in specific function (data not shown); (6) when afterload was increased but preload was unchanged, the improvement in cardiac function at high afterload was observed in the drug-treated diabetic group compared with the untreated diabetic group Significant improvement. While 50% of the untreated diabetic heart failed, about 90% of the triethylenetetramine-treated diabetic hearts still functioned well; (7) compared with untreated diabetic hearts, response to drugs Treated diabetic hearts showed improvements in a number of variables: improvements in cardiac output, aortic blood flow, coronary blood flow, and ventricular systolic and diastolic indices; (8) drug-treated groups of normal animals had no cardiac side effects; and (9) Histological observation (TEM and LCM) also showed the improvement of rat heart tissue structure after treatment with triethylenetetramine.

Treatment of STZ diabetic rats with triethylenetetramine significantly improved several measures of cardiac function. It can also be concluded that oral administration of triethylenetetramine for 7 weeks resulted in an improvement in overall cardiac function in 6-week-diabetic Wistar rats. This improvement has been demonstrated by an improvement in systolic function (+dP/dT) and a reduction in ventricular wall stiffness (-dP/dT). Triethylenetetramine also resulted in substantial improvement in the treatment of diabetic hearts resistant to increased afterload.

Example 7

This example was used to evaluate the effect of slow administration of triethylenetetramine on cardiac structure and function in diabetic and non-diabetic subjects.

Methods as below. Research on humans is sanctioned by institutional ethics and regulatory committees. The absorption and excretion of triethylenetetramine, and the representative plasma concentration-time curve after oral administration of triethylenetetramine have been reported (see MiyazakiK, et al., "Determination of trientine in plasma of patients with high-performance liquid chromatography , "Chem. Pharm. Bull. 38:1035-1038 (1990)).

Subjects (30-70 years old) who fill out the voluntary form are suitable for the study if they have the following conditions: HbA _Ic of T2DM >7%; cardiac ejection fraction (echocardiography) ≥ 45%, with obvious diastolic dysfunction , but no regional wall motion abnormalities; no other new drugs within 6 months; normal ECG (sinus rhythm, normal PR interval, normal T-wave and QRS configuration, and normal isoelectric st-segment ECG); and greater than 90% compliance with single-blind placebo treatment for a period of 2 weeks. Women were required to be postmenopausal, or infertile or non-lactating and non-pregnant with adequate contraceptive methods. If the subject does not meet the criteria, or has: morbidly obese (BMI ≥ 45kg.m ² ) Tl DM; obvious history of heart valve disease; obvious autonomic neuropathy; abnormal ventricular wall motion; history of multiple drug allergies; drug abuse; Abnormal laboratory tests; or MRI contraindications, etc., are not suitable for the experiment.

Prior to random sampling, potentially suitable subjects were enrolled in a 4-week single-blind phased treatment of two placebo capsules twice a day, and were screened by echocardiography, if regional wall motion abnormalities or impairment of LV systolic function (radiation blood fraction <50%), were excluded. In addition, assessment of LV diastolic filling was done using mitral inflow Doppler (reduced preload) to ensure that subjects had abnormal diastolic filling; normal subjects were not included. Subjects who met the criteria and had no reason to exclude were randomized to receive triethylenetetramine (600mg, twice a day, total dose 1.2gd ^-1 ) or two equivalent placebo capsules before eating, using double-blind parallel experiments . Assigned treatments were administered to maintain balance and were given to randomized subjects on a continuous basis. The double-blind treatment lasted 6 months.

At baseline and after 6 months of treatment, LV mass was determined using cardiac MRI, performed in the supine position using a phased array surface coil with the same 1.5T scanner (Siemens Vision). Apply segmented pulse sequence in 6 short-axis and 3 long-axis slices to obtain the expected gated cardiac cine image (TR 8ms; TE 5ms; flip angle 10°) with image sharing (11-19 frames.slice ^-1 ) ; field of view 280-350mm). Each slice was acquired during a breath-controlled period between 15-19 pulse beats. The short-axis slices spanned the left ventricle from top to bottom, the slices were 8 mm thick and spaced 2-6 mm apart. Long-axis slices were taken at approximately equal 60° intervals along the long axis of the left ventricle. Cardiac MRI provides accurate and reproducible assessment of LV mass and volume. The calculation of the mass and volume of the LV is carried out using the pilot point model, and the process can accurately measure the mass and volume. In short, the three-dimensional mathematical model of the LV is suitable for the simultaneous study of the epicardial and endocardial interfaces of the sliced LV wall, and then use the mathematical integration method to calculate the mass and volume of the model (mass = wall volume × 1.05g.ml ^{- 1} ). All measurements were done with data collected by one experimenter at the end of 6 months. Analysis of results was performed on hearts, applying maximum likelihood approximation to random data processing in a mixed model, and determining marginal least-squares-adjusted means. Changes from baseline were compared between treatment groups in a mixed model. Since only 2 groups had main effects and no interaction, such a procedure was not used. In additional analyses, the clinical impact of differences between treatment groups at baseline was considered intervariant with additional models. All P values were calculated for statistical significance by 2-tailed test and maintained at a 5% significance level. Treatment effects for defined variables were determined using the procedure of Mantel and Haenzel (SAS v8.01, SAS Institute).

Table 7 shows the basic information of 30 subjects with long-term type 2 diabetes. In a 6-month randomized, double-blind, placebo-controlled study of long-term oral triethylenetetramine dihydrochloride there was no clinically significant coronary artery disease or paradoxical diabetic function.

Table 7: Characteristics of Study Participants placebo Triethylenetetramine dihydrochloride N 15 15 average age 54 (range 43-64) 52 (range 33-69) female% 44% 56% average duration of diabetes 10(1-24) 8(1-21) Mean BMI (kg/m ² ) (SD) 32(5) 34(5) hypertension% 64% 80% Mean %HbA _1C (SD) 9.3(1.3) 9.3(2.0) Initial left ventricular mass (g) (SD) 202.2(53.1) 207.5 (48.7)

Triethylenetetramine (600 mg twice daily, for low-dose use in adult subjects with Wilson's disease, see Dahlman T, et al., "Long-term treatment of Wilson's disease with triethylene tetramine dihydrochloride(trientine), "Quart.J Med 88: 609-616 (1995)) or placebo given orally for 6 months in an equivalent diabetic adult group (n=15/group: Table 7), also combined with drug therapy including: β-receptor Blockers, calcium antagonists, ACE-inhibitors, cholesterol-lowering drugs, antiplatelet drugs and antidiabetic agents. LV mass by marker molecular response imaging (MRI see Bottini PB, et al., "Magnetic resonance imaging compared to echocardiography to assess left ventricular mass in the hypertensive patient," Am. J Hypertens 8:221-228 (1995)) at baseline and 6 Measured after three months of TET treatment. As expected, diabetes initially had significant LVH, consistent with previous reports. See Struthers AD & Morris AD, "Screening for and treating left-ventricular abnormalities in diabetes mellitus: a new way of reducing cardiac deaths," Lancet 359:1430-1432 (2002).

The results showed that triethylenetetramine reversed LVH in subjects with type 2 diabetes. Cardiac MRI scans at baseline and 6 months showed a significant reduction in LV mass. The mean value of the LV of the diabetic subject of the treatment of 6 months triethylenetetramine significantly reduces 5%, and placebo then has the improvement of 3% (accompanying drawing 33); LV mass keeps high for the index of body surface area significantly affected without changing systolic or diastolic blood pressure (Table 8). Thus, triethylenetetramine caused a strong regression of LV mass without altering blood pressure or urine output (Fig. 32). There were no significant drug-related side effects during the 6-month TET treatment period.

    Long-term triethylenetetramine treatment improves human cardiac structure and function

Table 8. Triethylenetetramine Treatment Outcomes placebo Triethylenetetramine treatment Urinary copper (μmol.l ^-1 ) 0.67(-1.16-2.49) -0.83(-2.4-0.74) Systolic blood pressure (mmHg) -1.9(-10.6-6.8) -3.5(-9.5-1.8) Diastolic pressure (mmHg) -4.5(-9.0-0.01) -3.9(-13.4-6.5) Left ventricular mass/body surface area (gm ^-2 ) +3.49(0.63-7.61) -5.56 ^** (-9.64-1.48)

Difference in primary treatment variable (6 months-baseline, mean (95% confidence interval ^* , P<0.05 vs. placebo ^** , P<0.01 vs. placebo).

MRI scans of the heart at baseline and 6 months showed a significant decrease in LV mass.

Briefly, triethylenetetramine administered for 6 months resulted in improvements in human heart structure and function.

Example 8

This example evaluates urinary metal excretion in diabetic and non-diabetic subjects chronically administered triethylenetetramine.

Methods as below. Research in humans was approved by the Public Ethics and Regulatory Committee. We measured urinary metal excretion in male subjects with T2DM or corresponding non-diabetic controls. Baseline information is given in Table 9 in a randomized, double-blind, placebo-controlled trial. Simple T2DM male subjects (Table 9) underwent a 12-day elemental balance study in fully associated metabolic units. All food and drink provided. Total daily intake (two-food method) and excretion (urine and feces) of trace elements (Ca, Mg, Zn, Fe, Cu, Mn, Mo, Cr and Se) were determined (ICP MS). Baseline measurements were performed during the first 6 days, after oral administration of triethylenetetramine (2.4 g, once daily) or the corresponding placebo in a 2×2 randomized double-blind trial, and metal loss was measured during the next 6 days.

Table 9: Characteristics of Study Participants placebo control Triethylenetetramine treatment control placebo diabetes group triethylenetetramine treatment diabetes group Average age n average diabetes age fasting glucose plasma (mmol.l ^-1 ) average HbA1C (%) body mass index (kg.m ^-2 ) 42(32-53)10-4.7±0.35.4±0.224.6±3.5 52(30-68)10-5.0±0.45.0±0.327.9±5.2 51(32-66)105.9(1-13)11.5±3.89.9±2.732.9±4.5 50(30-64)107.5(1-34)10.8±4.39.1±1.630.4±3.1

(Mean ± SEM, unless otherwise stated); fbg, HbA _1C and BMI were significantly higher in diabetes.

The results showed that the loss of urinary Cu was increased in type 2 diabetic subjects treated with oral triethylenetetramine. The urine output was the same in the drug group and the control group. Baseline 2-hour Cu loss was measured for the diabetic group (n=20) and the corresponding control group (n=20) within 10 hours of the first day (Figure 32); daily losses were measured on days 1-6 .

Baseline urinary Cu excretion was significantly higher in diabetics than in controls (mean 0.257 μmol.d ⁻¹ for diabetics, 0.196 for controls; P<0.001).

The 2-hour urinary Cu excretion of triethylenetetramine and placebo was measured again on day 7 in the same subjects (2.4 g once daily), with oral drug or placebo (n=10/group), three Ethylenetetramine increased urinary Cu in both groups, but the excretion rate was faster in the diabetic group (Fig. 30; P<0.05). Triethylenetetramine had no corresponding increase in Fe excretion, although basal concentrations were increased in diabetics relative to controls (P<0.001; results not shown). Thus, TET produced similar urinary Cu responses in T1DM rats and T2DM humans. Mean urinary Cu excretion by triethylenetetramine was 5.8 μmol.d ⁻¹ in T2DM compared to 4.1 μmol.d ⁻¹ in non-diabetic controls, a 40% increase. This agreement suggests that systemic Cu ^II increases are widespread in diabetic subjects.

Briefly, chronic administration of triethylenetetramine increased urinary copper levels in both diabetic and non-diabetic groups, but the rate of excretion was faster in the diabetic group. However, the application of triethylenetetramine had no corresponding increase in urinary Fe. Thus, triethylenetetramine produces a similar urinary copper response in rats with type 1 diabetes and human type 2 diabetes.

Example 9

This example was used to determine the effect of oral administration of triethylenetetramine (triethylenetetramine dihydrochloride) on the effect of metals on the fecal excretion of metals in diabetic and non-diabetic subjects. Methods as below.

Oral triethylenetetramine (2.4 g, once daily) or placebo was administered to the corresponding human with type 2 diabetes mellitus (T2DM) or the control group (n=10/group). Carry out studies of total metal balance in relevant metabolic units. Total defecation was collected daily for 12 days, freeze-dried, and analyzed by ICP-MS for Cu, Fe, Zn, Ca, Mg, Mn, Cr, Mb, and Se. Baseline measurements were performed on the first 6 days, and metal losses were measured on another 6 days in a 2 x 2 randomized double-blind trial following oral administration of triethylenetetramine or placebo.

The result is as follows. Mean daily fecal copper losses did not differ significantly between the triethylenetetramine and placebo groups, nor did Cu excretion differ between diabetic and control groups. The lack of effect of triethylenetetramine on fecal Cu excretion was unexpected (see Table 11), and is in marked contrast to reports for Wilson's disease, where increased fecal Cu excretion was reported.

Table 11 Excretion of fecal copper Average Cu loss (mg/day) Before treatment (Pre-Tment) After treatment (Post-Tment) Diabetes-Placebo (n=10) Control-Placebo (n=10) Diabetes-Drug (n=10) Control-Drug (n=10) 1.9145039651.6701421011.8698672932.19850868 1.9379212772.0786548921.9653423342.045467014 SEM: Diabetes-PrePlacSEM: Control-PrePlacSEM: Diabetes-PrePlacSEM: Control-PrePlacSEM: Control-PrePlac 0.1225703070.17657070.2282634650.209289978 0.1789957360.2094007860.1444630560.124516832 Reference Ishikawa et al. (2001): Control -1.00mg/d Kenzie Pamall et al. (1988): Control -1.30mg/d Kosaka H et al. (2001) control 53.5μg/d

Fecal excretion of other metals was similar. Neither diabetic nor non-diabetic subjects had detectable effects on excretion levels of Zn, Fe, Ca, Mg, Mn, Cr, Mb or Se. Briefly, triethylenetetramine did not increase fecal excretion of Cu or other metals in normal subjects and those with T2DM. Therefore, triethylenetetramine does not act by increasing fecal Cu excretion in T2DM. On the other hand, earlier studies showed that triethylenetetramine increased the urinary excretion of Cu. Taken together, these results suggest that triethylenetetramine can remove Cu from the body by increasing urinary copper loss. Therefore, triethylenetetramine in systemically active form is preferred in the present invention

Detailed ways.

Taken together, the human data and the rat data described above indicate that long-acting Cu chelation can cause significant regeneration of diabeticly injured hearts. Triethylenetetramine significantly reversed heart failure and LV damage in severely diabetic rats. Moreover, 6-month oral administration of triethylenetetramine significantly improved left ventricular hypertrophy in subjects with type 2 diabetes. Diabetic rats and humans have elevated systemic Cu ^II levels that can be treated with the Cu-selective chelator, triethylenetetramine.

Example 10

This example was used to evaluate the effect of parenterally administered different concentrations of a copper chelator in anesthetized male diabetic and non-diabetic Wistar rats by measuring urinary copper levels.

Stock solutions of various intravenous formulations containing various concentrations of triethylenetetramine dihydrochloride were prepared in 0.9% saline and stored at 4°C for 4 months without appreciable deterioration. The concentrations of the stock formulations were: 0.67 mg/ml, 6.7 mg/ml, 67 mg/ml, and 670 mg/ml. The formulation was then administered to rats at doses of 0.1 mg/kg, 1 mg/kg, 10 mg/kg, and t100 mg/kg, respectively.

After 6-7 weeks (mean, 44±1 days) of STZ administration, the animals were subjected to control or drug experiments. All animals were fasted overnight prior to surgery but were given deionized water ad libitum. Anesthetize with 3-5% trifluorobromochloroethane and ₂ ^l.min of O2. The femoral artery and vein were cannulated with a solid blood pressure transducer (MikrotipTM 1.4F, Millar Instruments, Texas, USA) and a saline-filled PE 50 catheter. The ureter was exposed by midline laparotomy, cannulated with a polyethylene catheter (0.9 mm external diameter, 0.5 mm internal diameter), and the wound was closed. Catheterized animals were ventilated with 70-80 breaths per minute and supplemented with _O2 (pressure-controlled ventilator, Kent Scientific, Connecticut, USA). Respiration rate and intratidal pressure (10-15 cm _H2O ) were adjusted to maintain intratidal _CO2 pressure at 35-40 mmHg (SC-300 _CO2 monitor, Pryon Corporation, Wisconsin, USA). During the operation and experiment, the body temperature was controlled at 37°C with a heating plate. Estimated fluid losses were supplemented intravenously with 154mmol.l ^-1 NaCl solution at a rate of 5ml.kg ^-1 .h ^-1 .

Mean arterial pressure (MAP), heart rate (HR, available from the MAP waveform), oxygen saturation (Nonin 8600V pulse oximeter, Nonin Medical Inc., Minnesota, USA) and core body temperature were monitored continuously throughout the experiment. Monitoring was performed using a PowerLab/16s data acquisition module (AD Instruments, Australia). The scaled signal is displayed on the screen and saved as two average values for each variable.

After 20 minutes of surgery and stabilization, experiments were started. Triethylenetetramine was administered intravenously over 60 seconds at increasing concentrations (0.1, 1.0, 10 and 100 mg.kg ^-1 in 75 μl of saline with 125 μl of saline flush). The control group received an equal volume of saline. Urine was collected during the experiment at 15-minute intervals.

Sample preprocessing was performed as follows. Urine: Urine was collected in pre-weighed 1.5 ml microtubes (eppendorf). The urine samples were reweighed, centrifuged and the supernatant diluted 25:1 with 0.02M 69% Aristar grade _HNO3 . Samples were stored at 4°C prior to GF-AAS analysis. If samples need to be stored for longer than two weeks, lyophilize and keep at -20°C. Serum : Terminal blood samples were centrifuged and serum was stored as urine until analysis. Trace metals in serum in blood samples were collected at the end of the experiment, and the renal clearance was calculated using the following equation:

Do the following statistical analysis. All values are expressed as ±SEM mean and P-value <0.05 indicates statistical significance. Unpaired t-tests were initially used to detect differences in weight and glucose between diabetic and control groups. To compare drug responses, statistical analysis was performed using analysis of variance (statistical values, see Windows v.6.1, SAS Institute Inc., Calfornia, USA). Subsequent statistical analysis was performed using a mixed model with repeated detection analysis of variance ANOVA (see Example 4).

The result is as follows. For the cardiovascular system, acute infusion of triethylenetetramine did not produce adverse effects. Figure 25 shows no adverse cardiovascular effects following infusion, although a transient drop in blood pressure occurred at 1000 mg/kg. The maximum drop in blood pressure during this change was 19±4 mmHg, but rats recovered within 10 minutes (not shown).

Briefly, acute intravenous administration of triethylenetetramine at concentrations ranging from 0.1 mg/kg, 1 mg/kg, 10 mg/kg, and up to 100 mg/kg had no significant effect on blood pressure. Furthermore, the triethylenetetramine formulation was effective as a copper chelator when administered intravenously, and triethylenetetramine maintained copper chelator activity in saline at 4°C for 4 months.

Example 11

This example evaluates the stability of triethylenetetramine doses with respect to copper sequestration capacity after storage.

Standard 100 mM triethylenetetramine HCl solutions were prepared in deionized water (MilliQ). Samples were stored in the dark at 4°C and in the dark at 21°C and a third sample was stored at 21°C in daylight.

The UV-Vis spectra of the formulations were initially measured on day 0, and then 20 μl aliquots of sample solutions were taken on day 15. Aliquots of 960 μl 50 mM TRIS buffer and 20 μl standard copper nitrate (100 mM—Orion Research Inc) were added to each sample. The binding stability of the triethylenetetramine formulations was determined at a wavelength of 700-210 nm. Referring to Figure 26, there is illustrated no appreciable change in copper chelation capacity of the formulation over a 15 day period. Furthermore, there is no detectable adverse effect on chelation under sunlight at room temperature, and triethylenetetramine is stable in solution as a copper chelating agent.

***

All patents, publications, scientific articles, network information or other documents and materials referenced or mentioned here are those skilled in the art that can be introduced into the present invention, and each referenced document and material is cited in its entirety. Here for reference. Applicant reserves the right to incorporate into the specification any and all optional materials and information from the above patents, publications, scientific articles, web information, electronic information, and other references or documents.

What this patent says includes all claims. Furthermore, all of these, including the original claims and claims from any and all priority texts, are hereby incorporated by reference in their entirety into the specification portion of this application, and applicants reserve the right to incorporate any and all such claims into this the descriptive or any other part of the specification of the application.

Claims are interpreted in accordance with the law. In no event, however, shall any adjustment or alteration of any claim, or part thereof, be made in the enforcement of this patent by reason of the alleged or perceived ease of understanding thereof .

All features of the description can be combined in any form. Thus, unless stated otherwise, each disclosed feature is only one example of an equivalent or similar feature.

It should be understood that the foregoing detailed description is used to demonstrate but not to limit the protection scope of the present invention, and the scope is defined in the form of claims. Therefore, in summary, it should be understood that although specific embodiments are used for illustration, various corresponding changes will not deviate from the spirit and scope of the present invention. Other aspects, advantages and improvements of the present invention are within the protection scope of the following claims. The invention is not to be restricted in any way except as described in the specific claims.

The specific methods and compositions described herein are illustrative of preferred embodiments and are not intended to limit the scope of the invention. Other themes, aspects and implementations can be obtained by those skilled in the art according to the specification, and are also included in the spirit and protection scope of the present invention. Various substitutions or modifications will be apparent to those skilled in the art without departing from the spirit and scope of the invention. The invention can illustratively be practiced in the absence or limitation of certain elements, which are not specifically described. Therefore, in the present invention, the terms "comprising", "including", "comprising" and the like are open-ended limitations. The process of the method of the present invention may also be practiced in a variable sequence of steps and does not require specific limitations.

The terms and expressions described here are descriptive and non-limiting, and do not exclude any stated equivalent features, and it should be recognized that various possible modifications are also within the protection scope of the present invention. Therefore, it is to be understood that although the present invention has specifically disclosed some preferred embodiments and optional features, all modifications and changes will occur to those skilled in the art and are included within the scope of the claims of the present invention. Inside.

This disclosure has described broadly and generally. Each specificity and sub-concept falling within this general disclosure may form a part hereof. This includes the general description of the invention conditioned and negatively limited excluding any inventive material from the general scope, whether or not said material is specifically mentioned herein.

It will also be understood that the singular forms of the terms "a" and "the" mentioned herein include plural forms unless specifically stated otherwise. The term "X and/or Y" means "X" or "Y", or "X" and "Y", and nouns include both their singular and plural forms. In addition, features or aspects of the present invention are described in terms of Markush groups, and those skilled in the art may recognize that the present invention is also described in individual forms or subgroups of Markush groups.

Other specific embodiments are contained in the claims. The patentability of the invention is not to be limited by any particular example or implementation or by any particular method and/or disclosure herein. Nor is it bound in any way by any representation of any examiner in the Patent, Trademark Division, or any other official or employee, unless such representation is specifically stated in the description of the specification of this application.

Claims (83)

1. A treatment method for preventing or improving tissue damage in a subject, wherein the subject does not suffer from Wilson's disease, the method comprising parenterally administering to the subject a therapeutically effective amount of copper Chelating agent, its range is about 5mg-1100mg. 2. The method according to claim 1, wherein said copper chelator is administered to said subject via parenteral route to treat diabetic heart disease, diabetic acute coronary syndrome, diabetic hypertension Cardiomyopathy, acute coronary syndrome associated with impaired glucose tolerance, acute coronary syndrome associated with fasting glucose impairment, hypertensive cardiomyopathy associated with impaired glucose tolerance, hypertensive cardiomyopathy associated with fasting glucose impairment A disease, symptom or disorder of cardiomyopathy, ischemic cardiomyopathy associated with impaired glucose tolerance, or ischemic cardiomyopathy associated with fasting glucose impairment. 3. The method of claim 1, wherein said copper chelator is administered parenterally to said subject to treat ischemic cardiomyopathy selected from the group consisting of myocardial infarction, coronary atherosclerosis , cardiomyopathy, myocarditis, idiopathic cardiomyopathy, metabolic cardiomyopathy, alcoholic cardiomyopathy, drug-induced cardiomyopathy, ischemic cardiomyopathy and hypertensive cardiomyopathy, acute coronary syndrome unrelated to glucose metabolism Hypertensive cardiomyopathy unrelated to glucose metabolism, or ischemic cardiomyopathy unrelated to glucose metabolism, disease, symptom or disorder. 4. The method of claim 1, wherein said copper chelator is administered parenterally to said subject to treat one or more diseases, conditions or disorders selected from the group consisting of aortic Disease, carotid artery disease Tree vascular disease, including cerebral vessels, coronary vessels, renal vessels, retinal vessels, iliac vessels, femoral vessels, popliteal vessels, vasa vasa, arteriolar tree and capillary beds Arterial disease, including Atherosclerotic disease of the major vessels of the aorta, coronary, carotid, cerebral, renal, iliac, femoral, or popliteal arteries. 5. The method of claim 1, wherein said copper chelator is administered to said subject parenterally to treat a disease, condition or disorder selected from cardiac structural damage. 6. The method of claim 5, wherein said structural damage to the heart is selected from atrophy, loss of cardiomyocytes, dilation of the extracellular space, or increased deposition of extracellular matrix. 7. The method of claim 1, wherein said copper chelator is administered parenterally to said subject to treat a disease, condition or disorder selected from structural damage to coronary arteries. 8. The method of claim 7, wherein the structural damage to the coronary arteries is selected from damage to the medial structure or the intimal layer. 9. The method of claim 1, wherein said copper chelator is administered parenterally to said subject to treat atherosclerotic plaque rupture involving one or more major blood vessels disease, condition or disorder. 10. The method of claim 9, wherein the major vessel is selected from the group consisting of the aorta, coronary, carotid, cerebral, renal, iliac, femoral, or popliteal arteries. 11. The method of claim 1, wherein said copper chelator is administered parenterally to said subject to treat symptoms including systolic dysfunction, diastolic dysfunction, systolic disturbances, recoil features A disease, condition, or disorder in which the disorder or function of the ejection phase is disturbed. 12. The method of claim 1, wherein said copper chelator is administered parenterally to said subject for the treatment of a disease, condition or disorder comprising microvascular disease. 13. The method of claim 12, wherein said microvascular disease comprises one or more of the group consisting of retinal arterioles, glomerular arterioles, vasa vasa, myocardial arterioles and associated ocular kidneys, heart, A disease of the blood vessels of the capillary bed of the central or peripheral nervous system. 14. The method of claim 1, wherein the copper chelator is administered in a single dose or in divided doses. 15. The method of claim 1, wherein said about 5 mg to about 1100 mg of said copper chelator is administered in single or divided doses per day. 16. The method of claim 1, wherein said copper chelating agent is triethylenetetramine. 17. The method of claim 1, wherein said copper chelating agent is a salt of triethylenetetramine. 18. The method of claim 1, wherein said copper chelating agent is a derivative of triethylenetetramine. 19. The method of claim 1, wherein said copper chelating agent is an analog of triethylenetetramine. 20. The method of claim 1, wherein said copper chelator is a prodrug of triethylenetetramine. 21. The method according to any one of claims 1-20, wherein said administration is by a method selected from the group consisting of transdermal administration, topical application, suppository administration, mucosal administration, inhalation administration, spray administration, buccal administered by sublingual, sublingual, or ophthalmic routes. 22. The method of any one of claims 1-20, wherein said administering is by injection. 23. The method of claim 22, wherein said injection is subcutaneous injection, intramuscular injection, implant administration or intravenous injection. 24. The method of claim 23, wherein said intravenous injection is a bolus injection or an intravenous drip. 25. The method of any one of claims 1-20, wherein said administering is accomplished by an infusion set. 26. The method of claim 25, wherein the infusion set is a passive infusion or an active implanted infusion set. 27. A delivery dosage unit or dosage formulation comprising a therapeutically effective amount of a copper chelator and a pharmaceutically acceptable delivery vehicle, said administration dosage unit being capable of delivering to a subject more than 10% w/w of said Copper chelating agent. 28. The unit of claim 27, wherein the dosage unit is administered orally, and said administered amount exceeding 10% w/w is the amount transferred from the intestine to the systemic circulation after oral administration. 29. The dosage unit of claim 28, wherein the copper chelating agent is a triethylenetetramine active agent. 30. The dosage unit of claim 29, wherein the copper chelating agent is triethylenetetramine. 31. The dosage unit of claim 29, wherein the copper chelating agent is a pharmaceutically acceptable salt of triethylenetetramine. 32. The dosage unit of claim 29, wherein the copper chelator is a prodrug of triethylenetetramine. 33. The dosage unit of claim 29, wherein the copper chelating agent is an analog of triethylenetetramine. 34. The dosage unit of any one of claims 28-33, wherein the drug delivery vehicle is loaded with or incorporates at least one delivery agent to increase the amount of drug from the intestine of the subject into the systemic circulation. 35. The unit of claim 34, wherein at least one delivery agent is one or more synthetic and/or natural polymers. 36. The unit of claim 35, wherein at least one delivery agent is one or more bioadhesive polymers. 37. The unit of claim 35, wherein at least one delivery agent is one or more passive diffusion agents. 38. The unit of claim 35, wherein at least one delivery agent is one or more active transport agents. 39. The unit of claim 35, wherein at least one delivery agent is one or more facilitated active transport agents. 40. The unit of any one of claims 28-39, wherein the copper chelator is admixed with a drug delivery vehicle. 41. The unit according to any one of claims 28-40, which is an enterically coated, enterically implanted and/or enterically contained dosage form. 42. The unit of any one of claims 28-41 which is of slow release type. 43. The unit of claim 42, wherein the unit has an enteric coated slow release core. 44. The unit of claim 42 or 43 comprising a coreless capsule. 45. The unit of any one of claims 28-44, wherein greater than 50% of the therapeutically acceptable chelating agent dissolves from the dosage unit into the gastrointestinal tract within 12 hours. 46. The unit of any one of claims 28-45 which is a repeated action dosage form having a reservoir for hierarchical release. 47. The unit of any one of claims 28-46, wherein the amount of copper chelator in the dosage unit is less than 300 mg. 48. The unit of claim 47, wherein the amount of copper chelator in the dosage unit is less than 250 mg. 49. The unit of claim 48, wherein the amount of copper chelator in the dosage unit is less than 240 mg. 50. The unit of claim 49, wherein the amount of copper chelator in the dosage unit is 120-140 mg. 51. The unit of any one of claims 28-50, wherein more than 15% w/w of the copper chelator is capable of being released from the intestine. 52. A unit as claimed in claim 51, wherein more than 20% w/w of the copper chelating agent is capable of being released. 53. A unit as claimed in claim 52, wherein more than 25% w/w of the copper chelating agent is capable of being released. 54. The unit of claim 53, wherein more than 30% w/w of the copper chelating agent is capable of being released. 55. The unit or formulation of claim 27, which is administered parenterally. 56. The dosage unit or formulation of claim 55, wherein the therapeutically acceptable copper chelator is a triethylenetetramine active agent. 57. The dosage unit or formulation of claim 56, wherein the therapeutically acceptable copper chelator is triethylenetetramine. 58. The dosage unit or formulation of claim 56, wherein the therapeutically acceptable copper chelator is a salt of triethylenetetramine. 59. The dosage unit or formulation of claim 56, wherein the therapeutically acceptable copper chelator is a prodrug of triethylenetetramine. 60. The dosage unit or formulation of claim 56, wherein the therapeutically acceptable copper chelator is an analog of triethylenetetramine. 61. The unit of any one of claims 55-60, wherein the dosage unit is administered transdermally. 62. The dosage unit of claim 61 which is a transdermal patch, pad, pack or bandage capable of being adhesively or otherwise bonded to the skin of a subject. 63. The formulation according to any one of claims 55-60, which is a topical formulation. 64. The dosage unit or formulation of claim 61 , 62 or 63, wherein at least 20% of the copper chelator is releasable into the systemic circulation. 65. The unit of claim 55, wherein said dosage unit is for sublingual administration. 66. The dosage unit of claim 65, wherein at least 20% of the copper chelator is releasable into the systemic circulation. 67. The unit of claim 27, wherein said dosage unit is in the form of a suppository. 68. The dosage unit of claim 67, wherein at least 20% of the copper chelator is releasable into the systemic circulation. 69. The dosage formulation of claim 55, which is injectable. 70. The formulation of claim 69, wherein at least 50% w/w of the copper chelator is releasable into the systemic circulation. 71. The formulation according to claim 69 or 70, which is administered intravenously. 72. The formulation according to claim 69 or 70, which is administered subcutaneously. 73. The formulation according to claim 69 or 70, which is administered intramuscularly. 74. The dosage unit or formulation of claim 51 in the form of a depot implant or a depot injection. 75. The unit or formulation of claim 74, wherein at least 20% w/w of the copper chelator is releasable into the systemic circulation. 76. The unit or formulation of claim 28, which is deliverable by an inhalation device. 77. The unit or formulation of claim 76, wherein at least 20% w/w of the copper chelator is releasable into the systemic circulation. 78. The formulation of claim 51 which is an ophthalmically administrable formulation. 79. Use of the dosage unit or dosage formulation of any one of claims 27-78 in the method of any one of claims 1-26. 80. Use of a dosage unit or dosage formulation according to any one of claims 27-79 packaged with a label or insert for use in the treatment of a disease, condition or disorder according to any one of claims 1-26. 81. The dosage unit or dosage formulation of claim 80, wherein once daily administration is required. 82. Use of copper chelating agents and other materials in the manufacture of dosage units or dosage formulations as claimed in any one of claims 27-81. 83. The use as claimed in claim 82 for the treatment use as claimed in any one of claims 1-26.

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