A METHOD FOR TREATMENT OF METABOLIC DISORDERS AND METABOLISM
FIELD OF INVENTION
The present invention provides a new use for known compounds. More particularly, the present invention provides a method of treating or preventing certain metabolic disorders of human and animal metabolism, such as non-insulin dependent diabetes mellitus (NIDDM) by the ad- ministration of the compounds listed in Table 1, below and excess adiposity or obesity by the administration of the compounds listed in Table 2, below.
Other indications which may be treated by the subject method can include hyperglycemia, impaired glucose tolerance, hyperinsulinemia, insulin insensitivity, hyperamylinemia or hyperlipidemia.
BACKGROUND OF THE INVENTION
There are several metabolic disorders of human and animal metabolism, e.g., hyperglycemia, impaired glucose tolerance, hyperinsulinemia and insulin insensitivity, hyperamylinemia, excess adiposity, and hyperlipidemia. Some or all of the above disorders may occur in the following disease states: non-insulin dependent diabetes mellitus (NIDDM), obesity, hypertension and atherosclerosis.
Hyperglycemia is a condition where the blood glucose level is above the normal level in the fasting state, following ingestion of a meal, or during a provocative diagnostic procedure, e.g., a glucose tolerance test. It can occur in NIDDM as well as obesity. Hyperglycemia can occur without a diagnosis of NIDDM. This condition is called impaired glucose tolerance or pre-diabetes. Impaired glucose tolerance occurs when the rate of metabolic clearance of glucose from the blood is less than that commonly occurring in the general population after a standard dose of glucose has been orally or parenterally administered. It can occur in NIDDM as well as obesity, pre-diabetes and gestational diabetes.
Hvperinsulinemia is defined as having a blood insulin level that is above normal level in the fasting state, following ingestion of a meal or during a provocative diagnostic procedure. It can be seen in NIDDM or obesity and can be associated with or causal in hypertension or atherosclerosis. Hyperinsulinemia can occur without a diagnosis of diabetes. It may occur prior to the onset of NIDDM. Insulin insensitivity, also called insulin resistance, occurs when the insulin- dependent glucose clearance rate is less than that commonly occurring in the general population during diagnostic procedures such as a hyperinsulinemic clamp (See, e.g., DeFronzo, R. A. et al.. Am. J. Physiol. 232:E214-E233, (1979)) or a minimal model test. See, e.g., Bergman, R. N. et al., J. Clin. Invest 68:1456-1467 (1981). Insulin insensitivity is considered also to occur when the blood glucose concentration is higher than that commonly occurring in the general population after
intravenous administration of insulin (insulin tolerance test) or when the ratio of serum insulin-to glucose concentration is higher than that commonly occurring in the general population after a 10 16 hour fast Insulin insensitivity may be found in NIDDM or obesity and can also be associate with or causal to hypertension or atherosclerosis.
Hyperamylinemia is defined as having an abnormally high blood amylin level. Amylin is also known as diabetes associated peptide (DAP) and insulinoma associated polypeptide CLAP). Hyperamylinemia can be seen in NIDDM or obesity.
Excess adiposity can be seen in NIDDM associated with obesity as well as obesity without NIDDM. It is defined as a higher fat body mass-to-lean body mass ratio than that commonly occurring in the general population as measured by whole body specific gravity or other generally accepted means.
Hyperlipidemia is defined as having an abnormal level of lipids in the blood. Hyperlipidemia exists when the serum concentration of total cholesterol or total triglycerides or the serum concentration of LDL-cholesterol/HDL-cholesterol is higher than that commonly occurring in the general population. It can be seen in NTDDM or atherosclerosis.
The above disease states could be treated by either ameliorating or preventing the metabolic and biochemical disorders. In addition, humans and animals, which have not been diagnosed as having one of the above disease states but evidencing some or all of the disorders described above, could be benefitted by preventing the development of a currently recognized disease state. Therefore, a compound that is useful in the treatment of hyperglycemia, impaired glucose tolerance, hyperinsulinemia, insulin insensitivity, hyperamylinemia. excess adiposity or hyperlipidemia could also be used to treat or prevent NIDDM, obesity, hypertension or atherosclerosis.
The subject invention provides a method for preventing or treating NIDDM using Compounds 1-119, listed in Table 1, their free bases, or their pharmacologically acceptable esters and salts. Compounds 1-119, their free bases, or their pharmacologically acceptable salts may be administered individually as the sole active ingredient in a composition or combined with other compounds selected from Table 1. Compounds 1-119 are known compounds and their sources are identified in Table 1.
The dose of Compounds 1-119 to be used is between 0.1 and 500 mg/kg body weight daily. The preferred dose is 1-50 mg/kg/day. Compounds 1-119 may be administered orally, buccally, sublingually, parenterally, intranasally, intrarectaliy, or topically in any suitable pharmaceutical formulation. The oral route is preferred.
The subject invention also provides a method of preventing or treating the obesity using
Compounds 1-128, listed in Table 2. their free bases, or their pharmacologically acceptable esters and salts. Compounds 1-128, their free bases, or their pharmacologically acceptable salts may be administered individually as the sole active ingredient in a composition or combined to form a
composition.
The dose of Compounds 1-128 to be used is between 0.1 and 500 mg/kg body weight daily. The preferred dose is 1-50 mg/kg/day. Compounds 1-128 may be administered orally, buccally, sublingually, parenterally, intranasally, intrarectally, or topically in any suitable pharmaceutical formulation. The oral route is preferred.
INFORMATION DISCLOSURE STATEMENT
Guanidine, monoguanidine and diguanidine compounds have been shown to produce hypoglycemia. See, e.g., Watanabe, C, J. Biol. Chem. 33:253-265 (1918); Bischoff, F. et al., Guanidine structure and hypoglycemia 81:325-349 (1929). However, these compounds were observed to be toxic. In 1957, biguanide derivatives, e.g. phenformin and metformin, were used clinically as anti-diabetic agents. Some members of this class continue to be used today while others have been withdrawn from the maricet or banned in the United States and most Western countries. See, e.g., Schafer, G., Diabete Metabol. (Paris) 9:148-163 (1983).
Gamma-guanidinobutyramide also known as Tyfoπnin, and the HC1 salt of Tyformin, known as Augmentin, were investigated as potential anti-diabetic agents from the mid-1960's until the mid-1970's. While Augmentin produced hypoglycemia, it was reported to produce hypertension in dogs [See, e.g., Malaisse, W. et al., Horm. Metab. Res. 1:258-265 (1969)] and respiratory and circulatory collapse in rats and rabbits. See, e.g., Buckle, A. et al., Horm. Metab. Res. 3:76-81 (1971). The free acid of the amide was said to lack hypoglycemic activity [See, e.g., Beeson, M. et al.. Horm. Metab. Res. 3:188-192 (1971)].
British patent 1,153,424 discloses the use of certain esters and amides of guanidino-aliphatic acids in the treatment of diabetes mellitus where hyperuremia is present The patent does not disclose that these compounds have an effect on hyperglycemia or any other symptom or pathological state related to diabetes. In a Canadian patent, 891509, the use of esters and amides of guanidinoaliphatic acids were disclosed for treating hyperuremia and hyperglycemia in diabetes mellitus. As noted above, the biologic activity of a guanidino alkanoic acid was known to be different and less favorable so as to be ineffective compared to its amide for treating hyperglycemia.
British patent, 1,195,199 discloses the use of guanidino alkanoic acids or their amides or esters in an insulin-containing, parenterally-administered composition for the treatment of hyperglycemia occurring in diabetes. According to this patent, the combining of a guanidino alkanoic acid, amide or ester with insulin reduces the risk of hypoglycemia as compared to insulin alone. British patent 1,195,200 discloses the use of guanidino alkanoic acids in a composition containing a guanidino alkanoic acid amide or ester derivative for the treatment of hyperglycemia occurring in diabetes. In a subsequent British patent, 1,552,179, the use of guanidino alkanoic acids, their salts, amides or esters in combination with a gluconeogenesis inhibitor for treating
hyperglycemic conditions was disclosed. Metformin was cited as an inhibitor of gluconeogenesis. Biological data indicated that HL 523, the preferred guanidino alkanoic acid derivative, was inactive as a single agent in six of seven experiments where blood glucose concentration was measured in alloxan diabetic mice and only weakly active in the seventh study. Most notably, British patents 1,195,199, 1,195,200 and 1,552.179 do not claim utility for guanidino alkanoic acids, as the sole active component in compositions for treating hyperglycemic symptoms in diabetes. Among the guanidino alkanoic acids tested, several were inactive as a single agent Thus, a variety of guanidino alkanoic acids lack significant anti-diabetic activity and combination of these compounds with an agent of known anti-diabetic activity, e.g., metformin, is necessary to show beneficial activity.
Aynsley-Green and Alberti injected rats intravenously with 3-GPA. argiriine, guanidine.4- guamdinobutyramide, and 4-guanidinobutyric acid. Arginine and 3-GPA stimulated insulin secretion transiently, but did not affect the blood glucose concentration while the other compounds stimulated insulin secretion but produced a rise in blood glucose concentration. See, e.g., Aynsley- Green, A. et al., Horm. Metab. Res. 6:115-120 (1974). Blachier, et al., observed that 10 mM 3- GPA stimulated insulin secretion by isolated rat islets in vitro. See, e.g., Blachier, F. et al„ Endocrinology 124:134-141 (1989). The insulin response induced by 3-GPA was 55% of that occurring when arginine was tested at the same concentration. In rats fed a diet supplemented with 10 mg/g 3-GPA for 30-60 days, the heart glycogen content was increased. See, e.g., Roberts, J. et al.. Am. J. Physiol. 243:H911-H916 (1982). Similarly, skeletal muscle glycogen content was increased in rats fed chow supplemented with lOmg/g of 3-GPA for 6- 10 weeks. Mice fed a diet supplemented with 3-GPA at 20 mg/g and supplied with drinking water containing 5 mg/ml 3-GPA for 7-12 weeks had serum glucose concentrations that did not differ significantly from mice receiving unsupplemented chow and water. See, e.g., Moerland, T. et al.. Am. J. Physiol.257:C810- C816 (1989).
With respect to adiposity, it is known that in some, but not all cases [See, e.g., Shoubridge,
E. et al., Biochem. J. 232:125-131 (1985)], supplementation of the diet with 10-20 mg/g 3-GPA results in decreased body weight See, e.g., Moerland, supra and Mahanna, D. et al., Exper. Neural. 68:114-121 (1980). This effect has been attributed to decreased skeletal muscle mass and has not been attributed to reduced adiposity or decreased lipid storage. See, e.g., Mahanna, supra and Shields, R. et al., Lab. Invest 33:151-158 (1975).
What is needed in the art is a sole therapy to treat or prevent the underlying metabolic disorders in these conditions.
SUMMARY OF THE INVENTION
In one aspect, the present invention provides a method of treating or preventing the metabolic disorder of NTDDM by administering to an animal exhibiting diabetes, including humans, an effective amount of a compound of Table 1 or a pharmaceutically acceptable salt thereof. Other
indications for which these compounds may be useful can include hyperglycemia, impaired glucose tolerance, hyperinsulinemia, hyperamylinemia, excess adiposity and/or hyperlipidemia. The method comprises the systemic administration of Compounds 1-119, listed in Table 1, their free bases, or their pharmacologically acceptable esters and salts to animals, including humans, suffering from NIDDM.
In another aspect, the present invention provides a method of treating or preventing a metabolic disorder such as excess adiposity or obesity in a patient susceptible to or experiencing said disorder comprising the systemic administration of Compounds 1-128, listed in Table 2, their free bases, or their pharmacologically acceptable esters and salts.
DETAILED DESCRIPTION OF THE INVENTION
Table 1, Compounds 1-119, their free bases, or their pharmacologically acceptable salts may be administered individually as the sole active ingredient in a composition for treating non-insulin dependent diabetes mellitus.
The Table 1 compounds 1-119 of mis invention are either commercially available or may be prepared by methods published in the chemical literature as indicated below in Table 1.
The Table 2 compounds 1-128, their free bases, or their pharmacologically acceptable salts may be administered individually as the sole active ingredient in a composition.
The Table 2 compounds 1-128 of this invention are either commercially available or ma be re ared bv methods ublished in the chemical literature as indicated below in Table 2.
The subject compounds cause several biologic effects that are beneficial in the treatment of human disease. They improve plasma glucose level, insulin sensitivity, plasma amylin level, adiposity and plasma lipid level. All of these effects are beneficial in treating metabolic disorders or metabolism such as NIDDM and excess adiposity or obesity.
NIDDM is characterized by hyperglycemia in the fasting or post-prandial state and impaired glucose tolerance after oral or parenteral administration of a glucose solution. The subject compounds, that are administered to KKAy mice, a rodent model of NIDDM, decreases the non-fasting plasma glucose concentration and improves glucose tolerance. The minimum effective dose in KKAy mice is 130 mg/kg/d when administered as an admixture in rodent chow. Higher doses produce a proportionately greater effect. Doses that are less than the minimum effective dose in KKAy mice may be effective at decreasing blood glucose levels in other species, e.g., human, since elimination is rapid in rodents and may occur more slowly in other species.
Impaired tissue insulin sensitivity and hyperinsulinemia occur in NIDDM [See, e.g., Defronzo, R., Diabetes 37:667-687 (1988) and Reaven, G., Diabetes 37:1595-607 (1988)], hypertension (See, e.g., Reaven, supra), obesity (See, e.g., Glass A., supra), and atherosclerosis [See, e.g., Reaven, supra and Stout, R. W., Diabetologia 16:141-150 (1979)] and may be etiological factors in these diseases. 3-GPA ameliorates hyperinsulinemia in KKAy mice and decreases the plasma ratio of insulin-to-glucose concentration, indicating increased insulin sensitivity. Therefore, 3-GPA is useful in the treatment or in the prevention of NIDDM, hypertension, obesity, and atherosclerosis.
Hyperamylinemia may occur in NIDDM, decreasing tissue glucose metabolism [See, e.g., Leighton, B. et al., Nature 335:632-635 (1988)] and altering pancreatic hormone secretion [See, e.g., Clark, A., Diabetic Medicine 6:561-567 (1989)]. 3-GPA ameliorates
hyperamylinemia and therefore is beneficial in treating disease states in which plasma amylin concentration is increased.
Excess adiposity is an etiological factor in NIDDM and when extreme, represents a disease state in itself. The subject compounds decrease adiposity by decreasing the level of lipids stored in fat and liver tissue. The compounds are therefore beneficial in the treatment of obesity alone or in concert with NIDDM. The effect of the subject compounds is selective for lipid-rich tissues {e.g., epididymal fat and fatty liver of ob/ob mice) while muscle mass is unaffected or only minimally affected.
Increased serum low density lipoprotein (LDL) cholesterol concentration is an etiological factor in coronary artery disease. The subject compounds decrease LDL-cholesterol levels in spontaneously hyperiipidemic mice and therefore is useful in treating or preventing
hyperlipoproteinemia, atherosclerosis and coronary artery disease.
"Sole active pharmaceutical agent" means that the subject compounds or its salt, administered as claimed herein, is the only pharmaceutical agent in the composition.
"Patients susceptible to or experiencing a metabolic disorder," i.e., hyperglycemia. impaired glucose tolerance, hyperinsulinemia, insulin insensitivity, hyperamylinemia. excess adiposity and/or hyperlipidemia means a human or animal who exhibits said metabolic disorder and is merefore likely to exhibit one of more of the disease states described above. Such patients are readily diagnosed by a physician or veterinarian of ordinary skill. "Treatment" means the amelioration or total avoidance of the metabolic disorder as described herein.
"Prevention" means the avoidance of a currently recognized disease state, as described herein, in a patient evidencing some or all of the metabolic disorders described above.
For all of these purposes, any convenient route of systemic administration is employed, e.g., orally, parenterally. intranasally or intrarectally. In general, the preferred form of administration is orally.
Compositions containing the compounds may be administered in a sustained release formulation. "Sustained release" means a formulation in which the drug becomes biologically available to the patient at a measured rate over a prolonged period. Such compositions are well- known in the art
Since the subject compounds decrease body fat without affecting the lean mass, they are of great commercial benefit to the meat poultry, and fish producing industries in achieving its goal of producing leaner animal products. The subject compounds can be administered admixed in the diet of farm animals or as a pharmaceutical preparation such as an oral tablet or capsule, by injection, or by implantable sustained release devices thereby increasing the protein content of the carcass while decreasing its fat content. This would produce muscle tissue with less fat. This benefit of the subject compounds would also impact on the potential health to the meat poultry, and fish consuming public. The term "farm animals" is defined as animals which are raised for food production. The term includes, but is not limited to, such animals as cattle, poultry, fish, swine, and lamb.
The subject compounds increase exercise tolerance in normal mice. Thus the present invention may be useful in treating muscular dysfunction, such as post-poliomyelitis chronic muscle fatigue syndrome or muscular dystrophy, or in treating chronic muscular weakness associated with advanced age or chronic immobilization, or in increasing endurance and exercise in normal humans.
The subject compounds are also useful for improving the survival rate of mice maintained in a low oxygen environment and therefore is beneficial in treating or preventing disease states involving tissue hypoxia, e.g.. peripheral claudication and exercise intolerance in diabetic humans, and angina, myocardial infarction and stroke in diabetic and normal humans.
It is known that glucose-dependent protein crosslinking alters the tertiary structure of several proteins . This protein glycosylation may contribute to diabetic complication and complications of aging in non-diabetic humans, such as neuropathy, nephropathy, retinopathy, hypertension, and atherosclerosis. The subject compounds are useful to block protein glycosy- lation and therefore be of benefit in treating or preventing this reaction.
The dosage regimen for the subject compounds in accord with this invention will depend on body weight. Table 1 and Table 2 compounds in pharmaceutical dosage form, can range from 1-500 mg/kg/day. The preferred dose is 5-100 mg/kg/day. Any sustained released formulations can be used.
The Table 1 compounds were tested for effects that are beneficial in the treatment or prevention of NIDDM using one or more of three procedures.
Procedure 1: Compounds were administered orally to KKAy mice for 3 days. Compounds were mixed in the chow at 1-5 mg/g or unsupplemented chow was provided. The blood glucose concentration was determined before initiating treatment and on the third treatment day.
Compounds that cause a decreased in blood glucose concentration during the study period at any of the doses that was greater by 20% or more than the decrease in blood glucose level, if any, occurring in control mice were considered active. KKAy mice are rodent models of non-insulin dependent diabetes mellitus (Iwatsuka, H., Shino, A., and Suzuoki, Z.: General survey of diabetic features of yellow KK mice, Endocrinol. Japon. 17: 23-35, 1970).
Procedure 2: Compounds were administered orally to C57EL63-ob/ob mice for 4 days.
Compounds were mixed in the chow at 5 mg/g or unsupplemented chow was provided. The blood glucose concentration was determined before initiating treatment and on the fourth study day. Compounds that cause a decrease in blood glucose level during the study period that was greater by 20% or more than the decrease in blood glucose concentration, if any, occurring in control mice were considered active, oblob Mice are rodent models of non-insulin dependent diabetes mellitus (Coleman, D. L.: Diabetes-obesity syndromes in mice. Diabetes 31, Suppl. 1: 1-6, 1982).
Procedure 3: Compounds were tested for their ability to antagonize carrier mediated transport of 3-guanidinopropionic acid into rat brain synaptosomes. Rat brain synaptosoraes were prepared as described (Fjalland, B., Acta Pharmacol, et Toxicol. 42: 73-76, 1978). Synaptosomes were incubated in Krebs Ringer bicarbonate buffer with 5 mM glucose and 0.1% bovine serum albumin, pH 7.4, for 5 min at 25°C with test compounds at a concentration of 1 mM and [4- 14C]-3-guanidinopropionic acid. Compounds that decreased synaptosomal accumulation of [4-
14C]-3-guanidinopropionic acid by >20% were considered active. The ability of compounds to antagonize synaptosomal uptake of 3-guanidinopropionic acid was found to significantly correlate with the decrease in blood glucose concentration in KKAy mice using Procedure 1. Thus antagonism in this assay was considered to be predictive of anti-NIDDM activity.
Effect of test compounds from Table 1 on blood glucose concentration in KKAy mice was measured and is shown in Table 3. Data are shown as the ratio of post-treatment blood glucose levels in treated (T) and control (C) mice. T/C-values <0.80 are considered active. Compounds were tested using Procedure 1. Stage 1 indicates the compound was tested at 1 mg/g; Stage 2, at 2 mg/g; Stage 5, at 5 mg/g.
The effect of Table 1 compounds on blood glucose concentration in oblob mice was measured and is shown in Table 4. Data are shown as the ratio of post-treatment blood glucose levels in treated (T) and control (C) mice. T/C-values <0.80 are considered active. Compounds were tested using Procedure 2.
The effect of Table 1 compounds on synaptosomal uptake of [4-14C]-3- guanidinopropionic acid is shown in Table 5. Compounds decreasing [4-14C]-3- guanidinopropionic acid uptake by >20% (i.e., <80% of control value) are considered active. Compounds were tested using Procedure 3.
The Table 2 compounds were tested for effects that are beneficial in the treatment or prevention of excess adiposity or obesity using one or more of three procedures.
Procedure 1: Compounds were administered orally to KKAy mice for 3 days. Compounds were mixed in the chow at 1-5 mg/g or unsupplemented chow was provided. The body weight was determined before initiating treatment and on the third treatment day. Compounds that cause a decreased in body weight during the study period at any of the doses that was greater than the weight decrease, if any, occurring in control mice receiving unsupplemented chow were considered active. KKAy mice are rodent models of obesity and diabetes (Iwatsuka, H., Shino, A., and Suzuoki, Z.: General survey of diabetic features of yellow KK mice, Endocrinol. Japon. 17: 23-35, 1970).
Procedure 2: Compounds were administered orally to C57BL6J-ob/ob mice for 4 days.
Compounds were mixed in the chow at 5 mg/g or unsupplemented chow was provided. The body weight was determined before initiating treatment and on the fourth study day.
Compounds that cause a decreased in body weight during the study period that was greater than the weight decrease, if any, occurring in control mice receiving unsupplemented chow were considered active, oblob Mice are rodent models of obesity and diabetes (Cawthome, M. A.: The use of animal models in the detection and evaluation of compounds for the treatment of obesity, In: "Animal Models of Obesity", New York: Oxford University, pp. 79-90, 1979).
Procedure 3: Compounds were tested for their ability to antagonize carrier mediated transport of 3-guanidinopropionic acid into rat brain synaptosomes. Rat brain synaptosomes were prepared as described (Fjalland, B., Acta Pharmacol, et Toxicol. 42: 73-76, 1978). Synaptosomes were incubated in Krebs Ringer bicarbonate buffer with 5 mM glucose and 0.1% bovine serum albumin, pH 7.4, for 5 min at 25 °C with test compounds at a concentration of 1 mM and [4- 14C]-3-guanidinopropionic acid. Compounds that decreased synaptosomal accumulation of [4- 14C]-3-guanidinopropionic acid by >20% were considered active. The ability of compounds to antagonize synaptosomal uptake of 3-guanidinopropionic acid was found to significantly correlate with weight loss in KKAy mice using Procedure 1. Thus antagonism in this assay was considered to be predictive of anti-obesity activity.
The effect of the Table 2 compounds on body weight in KKAy mice was tested and is shown in Table 6. Table 2 compounds were tested using Procedure 1. The first value indicates the compound was tested at 1 mg/g; the second value, at 2 mg/g; the fifth value, at 5 mg/g, etc. Percent (%) change is the body weight percent change.
The effect of the Table 2 compounds on body weight in oblob mice was tested and the values are shown in Table 7. Compounds were tested using Procedure 2.
The effect of the Table 2 compounds on synaptosomal uptake of [4-14C]-3- guanidinopropionic acid was tested and is shown in Table 8. Table 2 compounds decreasing [4- 14C]-3-guanidinopropionic acid uptake by >20% (i.e., <80% of control value) are considered active. Table 2 compounds were tested using Procedure 3.