MXPA01010037A

MXPA01010037A - Compositions and methods for treatment of diabetes.

Info

Publication number: MXPA01010037A
Application number: MXPA01010037A
Authority: MX
Inventors: Randy H Ziegler
Original assignee: Randy H Ziegler
Priority date: 1999-04-05
Filing date: 2000-04-04
Publication date: 2003-07-14
Also published as: JP2002541116A; WO2000059522A1; AU4196400A; EP1169047A1; AU780233B2; BR0009592A; CA2368215A1

Abstract

Products extracted from Brickellia californica, and isolated flavonoids that include apigenin, luteolin, quercetin and dihydrokaemferol, which are purified from Brickellia californica and useful for the treatment of diabetes are described. The extracted products and the purified flavonoids can be used alternatively in the treatment of diabetes dependent and non-insulin dependent because these materials produce a significant decrease in blood sugar.

Description

COMPOUNDS AND METHODS FOR THE TREATMENT OF DIABETES 1. Field of the invention The present application relates to the field of natural products and more specifically to extracts and plant compounds useful for the treatment of diabetes. 2. Description of the related art Diabetes mellitus (diabetes of honey or sugar), a potentially devastating, complex disorder of glucose metabolism, which is now partially controlled by insulin injections and drugs, is increasing in frequency throughout the world. In the United States it is estimated that more than 10 million people have diabetes. The economic cost is billions of dollars reflecting the cost of treatment and REF: 133515 loss of productivity while the human cost in impaired function, progression to blindness, limb amputation, kidney failure and heart and vascular disease is immeasurable. While the hallmark of diabetes is high blood sugar with the concomitant excretion of sugar in the urine, the disease has two main variants: Type I or Juvenile Initiation (insulin dependent diabetes mellitus), and Types II or Adult Initiation (diabetes mellitus not dependent on insulin). These variations are called that because of the approximate time of their initiation, but initiation is not really determinative. In summary, the IDDM appears to be a modulated immune inversion of the disease in which the production of insulin deteriorates while the NDDM is a disorder in which the cells do not respond to insulin. Diabetes is recognized in the ancient literature of Egypt, China, and India. Johann Conrad Brunner made the first suggestion that diabetes could involve a pancreatic disorder in 1682. It was not until the 20th century, however, that the diabetic condition was clearly associated with insulin - either the formation and the secretion of insulin by the pancreas or the influence of circulating insulin on the cells of the body. Simple glucose sugar is a major source of energy for human cells. Glucose is required for growth, development, and for optimal maintenance of the central nervous system. The brain is an avid consumer of glucose in such a way that any significant collapse of blood glucose results in a concomitant collapse in the level of glucose in the brain with the resulting cessation of normal brain function (coma). The entry of glucose into the cells and the metabolism of glucose within the cells are critical to sustain the life of the human body. Insulin, a transport regulating hormone, controls the uptake and transport of glucose in cells either for energy production or for storage in them. Glucose enters the bloodstream of the digestive system. If the intracellular glucose level is too low or the blood glucose level is too high, insulin is released to mediate the uptake of glucose by the cells for metabolism or storage, respectively. If the level of glucose in the blood is too low, other hormones mediate the release of glucose from glycogen (a storage polymer similar to a starch). Thus, insulin is necessary for the homeostasis of glucose that is found in the proper metabolism of the body. The correct concentration of insulin in the blood is critical. A lack of insulin leads to coma and death from the metabolic problems caused by excessive sugar in the blood.

On the other hand, an excess of insulin results in a shock caused by the excessively low level of sugar in the blood. Similarly, if the cells do not respond appropriately to insulin, homeostasis is interrupted and excessively high levels of blood sugar result. When the sugar In the blood is not controlled result metabolic balances of high glucose levels that lead to ketosis and alterations that damage the pH of the blood while inadequate levels of glucose lead to lethargy and coma. Dietary drugs and periodic insulin injections are now used in an effort to control life-threatening blood glucose oscillations. It is now well established that the damage is caused by excessive glucose and is not directly due to lack of insulin. Excess glucose combines with hundreds of proteins essential for normal metabolism and thus damages the cellular machinery of the body. The excess of glucose in the blood is responsible for the great morbidity of diabetes. Diabetics often suffer from small blood vessel disease (microangiopathy) caused by thickening of the walls of the capillaries over time. As a secondary result, capillaries leak, leading to retinopathy and nephropathy. In common terms, diabetes leads to blindness and damage to the kidneys. In addition, hardening of the arteries in the body can also cause premature coronary rupture. Neuropathy also occurs in diabetics and causes loss of sensation in the lower extremities. Gangrene and subsequent amputation are common events resulting from vascular deterioration mediated by diabetes. Insulin is produced inside the pancreas by 1.5 million beta cells located in clusters that are known as Islets of Langerhans. Insulin is a protein of moderate size composed of two chains: an alpha chain of 21 amino acids and a beta chain of 30 amino acids linked to each other by disulfide bonds. There are many theories to explain the deterioration of the production of insulin by the pancreas that leads to the diabetic condition. Reference is made to a document entitled "Autoimmune Imbalance and negative double T cells associated with resistant, tending and diabetic animals", Hosszufalusi, N., Chan, E., Granger, G., and Charles, M.; J. Autoimmun, 5: 305-18 (1992). This document shows that inflammation of the pancreatic islets interrupts the production of insulin. Specifically, the beta cells that produce insulin in the pancreatic islets are destroyed by the immune attack. It is recognized that such destruction of beta cells is due to attack by several types of immune cells including NK cells (natural killer) and double negative (CD4- [3/25 + OX19 +] / CD8- [0X8 + 0X19 +]) T-lymphocytes . More research progress has been achieved in this area and reference is made to a document entitled "Quantitative phenotypic and functional analysis of islet immune cells before and after the initiation of diabetes in the rat BB", Hosszufalusi, N. , et al., 36: 1146-1154 (1993), where it was shown that double negative T cells (CD4- / CD8) increased up to 30% of the islet T cell population at the onset of diabetes. It was shown that the cytolytic behavior of these cells was specific to islet cell tissue. A document entitled "Clonal suppression and autoreactivity in extramymal cells CD4-CD8- (double negative) alpha / beta receptor cells", Prud'homme, G.J., Bocarro, D.C., and collaborators, J. Immunol. 147: 3314-8 (1991), discusses the deletion of the VB 16 gene in the known variable region and the associated cytokines by a blocking compound which corrects the metabolic iiabalance that results in self-reactive negative double T cells - cells that cause inflammation at the islets in the pancreas.

A corrective balance of cell types was proposed as follows: B cells > T cells (CD4 > double negative > CD8) > NK cells > macrophages It is also recognized that an autoimmune response in the activation of macrophages by double negative T cells, where activated macrophages then attack the cells of the body. When appropriate depletion of T cell clones in the thymus low, the negative double T cells escape and become potentially self-reactive clones. It has been theorized that the CD8 protein, expressed by most NK cells, can be modulated by the administration of monoclonal antibodies to reduce the incidence of diabetes. The administration of polyclonal antibodies directed towards the AGMI glycolipid of the NK cell also prevents the autoimmune destruction of the Islets. At a neurological level, it is believed that aldosterone, from the adrenal cortex, sets in motion a group of reactions on the surface of all the cells of the body's tissues to regulate the uptake and retention of sodium and to extrude potassium. Reduced levels of sodium in g serum and high potassium levels in the serum increase the secretion of aldosterone. The adrenal glands are influenced by the neurotransmitter dopamine, an adrenal suppressor and by the neurotransmitter serotonin, an adrenal stimulator; Low potassium levels impact the production of dopamine and, therefore, alter the secretion of aldosterone and cortisol. In addition, other factors are involved in the negative feedback of pituitary corticotropin to cortisol. These factors have been recognized as atrial natriourotic peptides, or excretory sodium hormones, which inhibit the secretion of aldosterone, sodium chloride, potassium, and phosphorus. It has also been recognized that there is interference from the ongoing inhibition of prolactin by the hypothalamus dopamine as can be seen during the invasion of the pituitary stem by pineal tumors. These factors may be involved in the immunological abnormalities that lead to insulin-dependent diabetes or in abnormal insulin responses in insulin-independent diabetes. In a document entitled "Autoimmune diseases associated with the expression of the K + channel in CD4- and CD-8 T cells," Chandy, K. G., Eur. J. Immunol. 20: 747-751 (1990), the impact of potassium on the cytotoxicity created by DN T cells is discussed. Similarly, bioamines and neuropeptides were discovered that functioned as neurotransmitters to neuromodulate the inhibition or stimulation of neurotransmission, ie, peptides or opioids. In such mechanisms, the hypothalamus synthesizes and secretes neurohormones directly from and through the nerve axon to the capillary network transported by the pituitary portal circulation to the anterior pituitary gland. A document entitled "The document of growth factors in pancreatic cancer", Korc, M., Surg. Oncol Clin N A. , 7: 25-41 (1998), explains how insulin stimulates the growth and proliferation of cells through a tyrosine-synthase-dependent pathway. Insulin, like growth factor I (RGF-I), is a mitogenetic polypeptide that regulates the progression of the cell cycle. IGF-I and insulin are heterotetrameric proteins that possess intrinsic tyrosine synthase activity. IGF-I actions are dependent upon binding to their own specific cell surface receptors. Insulin and IGF-I activate the insulin receptor substrate -I (IRS-1), an important multi-site docking protein involved in mitogenic signaling. Activation of mitogenic pathways is magnified as a consequence of mutations in the K-ras oncogene and kinases associated with the cell cycle, such as pl6. Insulin exerts mitogenic effects on cells by activating the IGF-I receptor, which leads to the phosphorylation of IRS-1, an important regulatory protein that mediates growth promoting the effects of insulin. It is thought that tyrosine synthases truncate the dopamine production sequence thus causing a secondary receptor defect which has no affinity for the glucocorticoid necessary, but has affinity for DN4- and CD8- T cell proteins. It is theorized that this could be altered by proteoglycine to rebalance the K + channel (potassium) to allow a portal voltage to accumulate and allow the secretion of adequate amounts of aldosterone. It was also believed that an aggregated corrected series of polypeptide valencies assimilated into a proteoglycan would achieve this result. It is considered that diabetes is insidious, since until now no cure is known. However, several treatments have been used to improve diabetes. For example, dietary measures have been employed to balance the relative amounts of proteins, fats, and carbohydrates in the patient. In addition, diabetic conditions of moderate to severe intensity are treated with insulin administration. Also, prescription drugs such as "Glucoside" have been used to rejuvenate the impaired production of insulin in adult onset diabetes. Other drugs are used to modulate the effectiveness of insulin. In any case, in the treatment of diabetes, whether of the juvenile or adult initiation type, only partial success has been achieved.

BRIEF DESCRIPTION OF THE INVENTION According to the present invention there is provided a new and useful compound for treating diabetes. The treatment of the present invention uses a steam or aqueous extract of a plant known as Brickellia californica. The plant is harvested, dried, and combined with boiling water. The extract is then ingested orally by the patient periodically. It is known that the Brickellia genus is rich in flavonoids and other secondary products of the plant. The genus is broad and many species are mentioned in home medicine, in addition to B. californica, B. ambigens, B. arguta, B. brachyphylla. B. cylindracea, B. eupatoriodes, B. glutinosa, B. grandiflora, B. laciniata, B. lemmonii, B. oblongifolia, and B. veronicaefolia. Other species of the genus seem to have some or all of the active components of B. californica. Brickellia californica-specific flavonoids have been extracted and fractionated and given to diabetics with results similar to those produced by the extract. The specific flavonoids that were used were dihydrokaemferol and apigenin, a flavon. It was discovered then that these flavonoids are more effective in combination. Moreover, other Brickellia flavonoids, specifically myricetin and luteolin, have been determined to be effective in treating diabetes on their own or in combination, or in combination with dihydroxikaemferol and apigenin.

DETAILED DESCRIPTION OF THE PREFERRED MODALITY The following description is provided to enable any person skilled in the art to make and use the invention and the best models contemplated by the inventor to carry out his invention are set forth. However, several modifications will obviously be apparent to those skilled in the art, since the general principles of the present invention have been defined herein specifically to provide natural products extracted from Brickellia, and particularly flavonoids extracted from Brickellia, to treat diabetes . The following examples are illustrative of the invention but are not considered to limit the scope of the invention in any way.

Example I. Several living plants Brickellia californica were located and harvested. Brickellia is a small to medium sized shrub with relatively small, lobed leaves. Approximately four leaflets and stems were cut from the harvested plants. Each branch was approximately 20.32 cm (8 inches) in length. The twigs were placed in 1.89 liters (half a gallon) of water and heated to boil. It was continued to boil for 5 minutes, and then the extract was emptied from the container and cooled. The color of the drained liquid was light brown. The cold extract of the twigs of Brickellia californica was administered to four human male adults aged from thirty to 40 years. Each of these men suffered from diabetes. The dose for each subject was four to five glasses per day of the extract. Initially, all subjects were self-administering insulin at a level of 70 to 80 units per day. Blood glucose levels are measured periodically. After about three weeks, the glucose levels in each of the subjects began to drop. Subsequently, the insulin administered to the subjects was decreased. After approximately six weeks all subjects were able to control their diabetic condition without the use of exogenous insulin. The subjects suffered from adult onset diabetes and were using insulin because the antidiabetic drugs were not effective. At present, it is not known whether the Brickellia extract resulted in an increased production of insulin, in an increased function of insulin (for example, a higher or more efficient number of insulin receptors) or in a decrease in the blood by some mechanism that is not mediated by insulin. The material seems to be equally effective in cases of insulin-dependent diabetes. This could indicate that such diabetics have residual insulin production. As well, it is believed that the continued inflammatory destruction (discussed above) of beta cells continues in insulin-dependent diabetics. It seems very likely that the Brickellia extract modulates this process allowing the survival of beta cells and the production of insulin. It is also possible that the extract also increases the effect of residual insulin or works by another mechanism hitherto unknown.

Example II Live plants of Brickellia californica were harvested and dried. The material of the dried plant was ground using mortar and pestle, transferred to a 125 ml Erlenmeyer flask and extracted with a mixture of chloroform and methanol in a ratio of 1: 1. The mixture was beaten for 4 hours with a beater magnetic. The extract of the bottle was then filtered to remove the remains of the cells and concentrated in a "rotavap" under vacuum to produce a chewy crude residue. The residue was divided into chloroform and methanol to produce two fractions labeled CHC13 (the soluble fraction of chloroform plus hydrophobic) and MeOH (the soluble fraction of methanol plus hydrophilic). The CHC13 and MeOH fractions were analyzed using a Hewlett Pakcard 6890 gas chromatographic mass spectrometer (GC-MS) adapted with an HP-5MS capillary column (30 meters x 250 μ x 0.25 ym). The conditions of the analysis were as follows: The initial temperature was 125 ° C which was sustained for 5 minutes, followed by an increase to 275 ° C at a rate of 10 ° C per minute with the final temperature of 275 ° C sustained by 15 minutes. The analysis of the CHC13 fraction by GC-MS showed the presence of a group of polar flavonoids with retention times in a range of 13 to 15 minutes, the presence of a group of sesquiterpenes with retention times between 16 to 18 minutes, and a small group of aliphatic hydrocarbons with retention times between 20 to 25 minutes. The GC-MS analysis of MeOH fraction produced similar results except that the MeOH fraction was mostly free of aliphatic hydrocarbons. It is believed that the extract of Brickellia californica includes the flavonoids dihydroxikaemferol, apigenin, luteolin, myricetin and quercetin. In addition, the many other Brickellia species contain these, or similar flavonoids, although perhaps in different proportions, and should also be effective in the treatment of diabetes. Experiments with diabetic test animals (rats and mice) were carried out. The Brickellia extract was effective in controlling blood glucose in these model systems. In addition, the administration of synthetic versions of Brickellia flavonoids were also effective in lowering glucose levels. In treatments that involved a single flavonoid, luteolin was the most effective agent. However, there is some indication that a combination of luteolin with other flavonoids, especially dihydroxikaemferol and apigenin, results in an increased effect since the level of glucose in the blood can be lowered to the maximum with a lower flavonoid dose in general. The effect seems more pronounced when the molar concentration of luteolin is at least twice that of combined dihydrokaemferol and apigenin. Whatever the route of the action of the flavonoids, the results are not instantaneous. As explained above, the Brickellia extract takes a few weeks to lower blood glucose to the maximum. In model animals, it takes several days for an appreciable reduction of blood glucose, the maximum effect requires several weeks. This delay in the results may explain why this effect has not been observed previously considering the many common flavonoid-containing fruits and vegetables that have been shown to be effective in the present invention. It would appear that sustained ingestion of adequate amounts of effective flavonoids would be required. On the other hand, it is well known that the original human diets were rich in flavonoids while the common diets refined in the industrialized nations are relatively poor in flavonoids. Recent studies have suggested that the lack of dietary flavonoids is partially responsible for coronary and vascular diseases. Now it seems that the global "epidemic" of diabetes can also be a result of the lack of flavonoids. It is known that vegetarians have lower incidences of diabetes as well as a number of other degenerative diseases. The conventional wisdom is that the absence of diabetes could be related to the relative absence of refined sugars from the vegetarian diet. An alternative explanation could be the richness of flavonoids in these diets. In addition to the equivalents of the elements affirmed, obvious substitutions now known or known by a person skilled in the art are defined to be within the scope of the defined elements. Therefore the claims should be understood to include what is illustrated and described specifically above, what is conceptually equivalent, what can obviously be substituted, and also what essentially embodies the essential idea of the invention. Those skilled in the art will appreciate that the various adaptations and modifications of the preferred embodiment described above can be configured without departing from the scope of the invention. The illustrated embodiment has been set forth only for the purpose of example and should not be taken as a limitation of the invention.

It is noted that in relation to this date, the best method known to the applicant to carry out the aforementioned invention, is the conventional one for the manufacture of the objects or products to which it refers.

Claims

Having described the invention as above, the content of the following claims is claimed as property: 1. An antidiabetic composition characterized in that it comprises an extract of plants of the Brickellia genus. 2. An antidiabetic composition according to claim 1, characterized in that it comprises an extract of Brickellia californica.
3. An antidiabetic composition characterized in that it consists of a flavonoid selected from the group consisting of luteolin, myricetin, dihydrokaemferol, apigenin, quercetin and mixtures thereof.
4. An antidiabetic composition characterized in that it consists of a mixture of luteolin, dihydrokaemferol and apigenin.
5. The antidiabetic composition according to claim 4, characterized in that the mole concentration of luteolin is at least twice that of combined dihydroxikaemferol and apigenin.
6. A method for the treatment of diabetes mellitus characterized in that it comprises the step of administering an amount of an aqueous extract of plants of the Brickellia genus to produce a reduction of glucose in the blood.
7. The method according to claim 6, characterized in that the extract is from Brickeliia californica.
8. A method for the treatment of diabetes mellitus characterized in that it consists of the step of administering an amount of a flavonoid selected from the group consisting of luteolin, myricetin, dihydrokaemferol, apigenin, quercetin and mixtures thereof to produce a reduction in glucose in the blood.
9. The method according to claim 8, characterized in that a mixture of dihydrokaemferol and apigenin is administered.
10. The method according to claim 9, characterized in that the mole concentration of luteolin is at least twice that of combined dihydroxikaem erol and apigenin.