JP2002541116A - Compositions and methods for the treatment of diabetes - Google Patents

Abstract

  (57) [Summary] Extract products from Brichelia californica and isolated flavonoids including apigenin, luteolin, quercetin and dihydroxychemferol purified from Brichelia californica are useful for the treatment of diabetes. Extract products and purified flavonoids can be used for both insulin-dependent and non-insulin-dependent diabetes. This is because these materials provide a significant reduction in blood glucose.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION

The present invention claims priority from provisional application serial number 60 / 127,824, filed April 4, 1999, entitled "Compositions, Products and Methods for the Treatment of Diabetes". It is incorporated herein by reference.

[0002]

TECHNICAL FIELD OF THE INVENTION

The present invention relates to the field of natural products, and in particular to plant extracts and compounds useful for treating diabetes.

[0003]

[Prior art]

Diabetes mellitus (diabete) is a complex failure of glucose metabolism that can be severe.
s mellitus, honey or sugar diabetes) is increasing worldwide and is currently partially controllable by insulin injections and / or drugs. In the United States, 10
It has been estimated that over a million people have diabetes. Its economic costs are in the multi-billion dollars, reflecting loss of treatment costs and productivity, and human costs such as dysfunction, progression to blindness, limb amputation, kidney failure, and heart and vascular disease are high. It cannot be measured.

[0004] Diabetes is usually confirmed by the detection of sugars in urine due to high blood sugar levels, which include type I or juvenile diabetes (insulin-dependent diabetes mellitus); and II There are two major types, the type or adult diabetes (non-insulin dependent diabetes mellitus).

[0005] These types are named by the approximate time of onset of diabetes, but in practice, the onset of diabetes is not a critical factor. Simply put, IDDM
(Insulin-dependent diabetes mellitus) is one in which insulin secretion is reduced due to modulation of immunity, and NDDM (non-insulin-dependent diabetes mellitus) is a disorder in which cells do not respond to insulin.

[0006] Diabetes has also been recognized in ancient literature, such as in Egypt, China, and India, and Johann Conrad Brunner first suggested in 1682 that diabetes might involve pancreatic disorders. ing. However, it was only in the 20th century that the symptoms of diabetes, such as the formation and secretion of insulin by the pancreas or the effects of circulating insulin on cells in the body, were related to insulin.

[0007] Glucose is a monosaccharide, the main energy source of human cells, and is essential for successful growth, development, and maintenance of the central nervous system. The brain is a greedy consumer of glucose, and if blood glucose levels drop significantly, blood glucose levels in the brain will fall and normal brain function will cease (coma). It is necessary for the human body to sustain life if glucose enters the cells and metabolism occurs inside the cells. Insulin, a regulatory transport hormone, controls glucose uptake and transport into cells for energy production or storage. Glucose enters the bloodstream through the digestive system. If glucose levels between cells are too low or glucose levels in the blood are too high, insulin is released, which mediates the uptake of glucose by cells for metabolism and storage. If the glucose levels in the blood are too low, other hormones mediate the release of glucose from glycogen (a starch-like storage polymer). Therefore, insulin is necessary for glucose homeostasis in proper body metabolism. Proper concentration of insulin in the blood is an important factor. Lack of insulin can lead to coma and death from metabolic problems due to excessive blood sugar. On the other hand, an excess of insulin causes a shock symptom due to an excessively low blood sugar level. Similarly, if the cells do not respond properly to insulin, homeostasis is interrupted, resulting in an excessive blood glucose level.

[0008] If blood sugar levels are not controlled, metabolism becomes severely imbalanced, leading to elevated glucose levels leading to ketosis and dramatic changes in blood pH. On the other hand, insufficient glucose levels lead to lethargy and coma. Today, periodic infusions of diet drugs and / or insulin are being used to control life-threatening glucose fluctuations. Damage is due to excess glucose, not a direct cause of lack of insulin,
It is now well recognized. An excess of glucose mixes with a number of proteins required for normal metabolism, damaging the functioning of the body's cells in that way.

[0009] Excess of blood glucose is responsible for many diabetes pathologies. Many diabetic patients suffer from microangiopathy caused by the thickening of the capillary wall over time. A secondary consequence is that the capillaries can leak easily, leading to retinopathy and nephropathy. Similarly, diabetes can lead to blindness and kidney damage, and atherosclerosis in the body can lead to premature coronary rupture. In addition, a diabetic patient may have neuropathy, causing loss of lower limb sensation. The occurrence of gangrene leading to amputation is also a common consequence of diabetes-mediated vascular disorders.

[0010] Insulin is produced in the pancreas from 1.5 million β cells located in clusters called “islets of Langerhans”. Insulin is a medium-sized protein consisting of two chains, an α-chain of 21 amino acids and a β-chain of 30 amino acids, which are linked together by disulfide bonds.

There are a number of theories explaining that impaired insulin production by the pancreas leads to diabetes symptoms. "Autoimmune Imbalance and Double Negative T Cells Associ
ated with Resistant, Prone and Diabetic Animals ", Hosszufalusi, N., Chan
, E., Granger, G., and Charles, M .; J Autoimmun, 5: 305-18 (1992). The article explains that inflammation of islets in the pancreas interferes with insulin production. In particular, beta cells that produce insulin on islets in the pancreas are destroyed by immune attack. Such destruction of β cells can be caused by NK (natural killer) cells or double negative (CD4- [W3 / 25 + OX19 +] / CD
8- [OX8 + OX19 +]) has been recognized as being due to attack by several types of immune cells, including T lymphocytes.

[0012] Research in this area has further advanced and has been described in "Quantitative Phenotypic and Funct
ional Analyses of Islet Immune Cell Before and After Diabetes Onset in t
he BB Rat ", Hosszufalusi, N .. et al., Diabetologia 36: 1146-1154 (1993)
Is cited. Among them, double negative T cells (CD4- /
It has been demonstrated that CD8-) had increased to 30% of the islet T cell count at the onset of diabetes. The cytolytic behavior of these cells has been shown to be tissue-specific for islet cells.

[0013] Clonal deletion and autoreactivity in extrathymic CD4-CD8- (double ne
gative) T cell receptor-alpha / beta T cells ", Prud'homme, GJ, Bocarro,
In a paper entitled DC, et al., J Immumol. 147: 3314-8 (1991), a block compound that corrects a metabolic imbalance inhibits the known variable region gene VB16 and related cytokines, We are examining the resulting self-reactive double-negative T cells-cells that cause inflammation of islets in the pancreas. The types of cells that correct the balance are proposed to be: B cells> T cells (CD4> double negative>CD8)> NK cells> macrophages. It has also been recognized that the autoimmune response is due to the activation of macrophages by double-negative T cells, and that the activated macrophages attack body cells. I have. If T cell clone depletion in the thymus is not performed properly, double negative T cells escape and become clones that may have autoreactivity. It is theorized that the CD8 protein expressed by the majority of NK cells can be regulated by administration of monoclonal antibodies, which can reduce the incidence of diabetes. Administration of a polyclonal antibody against the glycolipid AGMI of NK cells also prevents autoimmune islet destruction.

At the neurological level, it is believed that aldosterone from the adrenal cortex drives a series of reactions to regulate sodium uptake and retention and potassium excretion at the surface of all cells of body tissue. I have. Reduced serum sodium and elevated serum potassium levels enhance aldosterone secretion. The adrenal gland is affected by the neurotransmitter dopamine, which is an adrenal depressant, and the serotonin, a neurotransmitter, which is an adrenal stimulator. Alter secretion. In addition, there are other factors in the negative feedback of pituitary corticotropin to cortisol. These factors have been identified as atrial natriuretic peptides or natriuretic hormones that interfere with the secretion of aldosterone, sodium chloride, potassium, and phosphorus. It has also been recognized that there is interference with the inhibition of prolactin caused by dopamine from the hypothalamus, as seen when the pituitary stalk is eroded by pineal tumors. These factors are involved in immune abnormalities and can lead to abnormal insulin responses in insulin-dependent or non-insulin-dependent diabetes.

[0015] "Auto Immune Diseases Linked to Abnormal K + Channel Expression in DN C
D4- and CD8- T cells ", Chandy, KG, et al, Eur. J. Immunol. 20: 747-75
1 (1990) discusses the effects of potassium on cytotoxicity caused by DNT cells. Similarly, biogenic amines and neuropeptides have been found to function as neurotransmitters that neuromodulate inhibition or stimulation of neurotransmission, ie, opioid peptides. In such a function, the hypothalamus synthesizes and secretes neurohormones directly and via nerve axons into the capillary network and is transported through the pituitary portal vein to the anterior pituitary gland.

"Role of growth factors in pancreatic cancer", Korc, M., Surg Oncol Cl
A paper entitled in N Am., 7: 25-41 (1998) describes how insulin stimulates growth and cell division through a tyrosine kinase-dependent pathway. Insulin, like growth factor I (RGF-I), is a mitogenic polypeptide and regulates cell cycle progression. IGF-I and insulin are heteroquadrant proteins and have endogenous tyrosine kinase activity. The action of IGF-I depends on its specific binding to cell surface receptors. Both insulin and IGF-I are insulin receptor substrates-I (
It activates IRS-I), an important multisite binding protein involved in mitogenic signals. Mutations in the K-ras oncogene and cell cycle-related kinases such as p16 result in enhanced mitogenic pathway activity. Insulin exerts a mitogenic effect on cells by activating the IGF-I receptor, leading to the phosphorylation of IRS-1, an important regulatory protein that mediates the growth promoting effect of insulin. Tyrosine kinases are thought to be truncating sequences that produce dopamine, resulting in post-receptor deficiencies that result in a complete loss of affinity for the required glucocorticoid, but a decrease in DNT cell CD4- and CD8-proteins. Comes with affinity. It is theorized that this can be altered by proteoglycine to rebalance the K + (potassium) channel, accumulate gate voltage and secrete the appropriate amount of aldosterone. It was also believed that a series of polypeptides with a corrected outer surface, aggregated, and mimicking proteoglycans would achieve this result.

[0017] Diabetes is considered to be an insidious disease for which no cure has yet been found. However, various treatments are used to ameliorate the disease. For example, a diet has been employed that balances the relative amounts of protein, fat, and carbohydrate in a patient. In addition, moderate to severe diabetes symptoms are treated with insulin administration. In adult diabetic patients, prescription drugs such as "glucoside" used for rejuvenating insulin production are used. Other drugs are used to regulate the way insulin works. In any case, the treatment of diabetes, whether young or adult, has not been completely successful.

[0018]

SUMMARY OF THE INVENTION

According to the present invention, there is provided a novel and useful composition for treating diabetes. The treatments of the present invention utilize a steam or aqueous extract of a plant known as Brickellia californica. The plants are collected, dried and mixed with boiling water. The extract is regularly administered orally to the patient. The genus Brichelia is known for its richness in flavonoids and other secondary plant products. This plant genus is large, and many species have been documented in folk remedies.
In addition to California, B. ambigens, B. argut
a), B. brachyphylla, B. cylindracea, B.
B. eupatoriodes, B. glutinosa, B. grandiflora, B. laciniata, B. remony
lemmonii), B. oblongifolia, and B. veronicaefolia. Other species of this genus are It appears to contain any or all of the active ingredients in California.

Certain flavonoids have been extracted and fractionated from B. californica and administered to diabetics, with results similar to those obtained with the extracts. The flavonoids used in particular are dihydrochemferol, apigenin and a flavone. At that time, it was discovered that these flavonoids were most effective when combined. In addition, it has been determined that other Brichelian flavonoids, especially myricetin and luteolin, alone or in combination, or in combination with dihydrochemferol and apigenin, are effective in treating diabetes.

[0020]

DETAILED DESCRIPTION OF EMBODIMENTS

The following description sets forth the best mode for carrying out the invention, which is provided to enable any person skilled in the art to make and use the invention and is intended by the inventor. However, since the general principle of the present invention is now defined as providing natural products extracted from Brichelia, particularly flavonoids extracted from Brichelia, and treating diabetes, various modifications may be made. It will still be readily apparent to those skilled in the art.

The following examples are illustrative of the present invention, but are not to be construed as limiting the scope of the invention in any manner.

Example I Several live trees of Brichelia californica were found and collected. Brichelia is a small to medium shrub with relatively small, shallow leaves. Approximately four leaf sprigs of leaves and stems were cut from the collected plants. Each twig was about three inches long. The twigs were placed in half a gallon of water and heated to boiling. After 5 minutes of boiling, the extract was decanted from the container and allowed to cool. The color of the decanted liquid was light brown. Extracts from chilled Brichelia californica twigs were administered to four adult men aged 30 to 40 years. Each man was diabetic. The dose for each subject was 4-5 glasses of extract per day. Initially, all subjects are 70-
He had given himself 80 units of insulin. Blood glucose levels were measured regularly, and after about three weeks, each subject's blood glucose levels began to drop. As a result, the amount of insulin administered to the subject also decreased. After approximately six weeks, all subjects were able to control their diabetes symptoms without exogenous insulin.

These subjects had adult diabetes and had been using insulin because conventional antidiabetic drugs had been shown to be ineffective. At present, whether the extract of Brichelia enhances insulin production, enhances insulin function (increases in number of insulin receptors, increases efficiency), or something mediated by something other than insulin It is not known whether the blood sugar level is lowered by the function. This material appears to have a similar effect in the case of insulin-dependent diabetes. This may indicate that such diabetic patients still have insulin production. Also, β-cell destruction due to continuous inflammation (described above) is thought to continue in insulin-dependent diabetic patients. It is likely that Brichelia extract regulates this process, allowing β cells to survive and produce insulin. This extract may enhance the effects of residual insulin or manipulate other unknown functions.

Example II. The live trees of Brichelia californica were collected and dried. The dried plant material is deflocculated using a mortar and pestle and transferred to a 125 ml Erlenmeyer flask, 1: 1
And extracted with a mixture of chloroform and methanol in the following ratios: The mixture was stirred with a magnetic stirrer for 4 hours. The extract from the flask was filtered to remove cellulose compound debris and concentrated under vacuum in a “rotavap” to give a crude gummy residue. This residue was partitioned between chloroform and methanol to give two fractions labeled CHCl ₃ (more hydrophobic soluble fraction) and MeOH (more hydrophilic methanol soluble fraction). Was.

The fractions of CHCl ₃ and MeOH were obtained from Hewlett-Packard 6890 gas chromatograph
A mass spectrometer (GC-MS) is used to connect an HP-5MS capillary column (3
(0 meter × 250 μm × 0.25 μm). The analysis conditions are as follows. The initial temperature was held at 125 degrees Celsius for 5 minutes, then the temperature was raised to 275 degrees Celsius at a rate of 10 degrees per minute, and the final temperature was maintained at 275 degrees Celsius for 15 minutes. Analysis of the CHCl ₃ fraction by GC-MS showed the presence of a group of polar flavonoids with a retention time of 13-15 minutes, the presence of a group of sesquiterpenes with a retention time of 16-18 minutes, and a retention time of 20-25 minutes. The presence of a small group of aliphatic hydrocarbons was indicated. Analysis of the MeOH fraction by GC-MS gave similar results,
The OH fraction was totally free of aliphatic hydrocarbons.

It is believed that the extract of Brichelia californica contains the flavonoids dihydroxychemferol, apigenin, luteolin, myricetin and quercetin. In addition, many other Brichelian species, although in varying proportions, contain these or similar flavonoids and should be effective in treating diabetes. Experiments with diabetic test animals (rats and mice) were performed. The extract of Brichelia was also effective in controlling blood glucose in these model systems. Also, administration of the synthesized Brichelia flavonoids also had the effect of lowering glucose levels. Luteolin was the most effective drug for treatment with a single flavonoid. However, the combination of luteolin with other flavonoids, especially dihydroxychemferol and apigenin, has been shown to provide enhanced effects that can lower blood glucose maximally at lower total flavonoid doses.

Whatever the route of action of the flavonoids, the results are not immediate. As explained above, it takes weeks for the extract of Brichelia to lower blood glucose to its maximum. In animal models, it can take days to see a drop in blood glucose and weeks to get maximum results. This delay in results is due to the fact that this effect has never before been observed, despite the fact that many common fruits and vegetables contain flavonoids that prove to be effective in the present invention. Could explain. It seems necessary to maintain an adequate intake of effective flavonoids. It is well known that flavonoids were originally abundantly contained in human diets, but flavonoids are relatively abated in sophisticated diets in industrialized countries. Recent studies suggest that dietary flavonoid deficiency is part of the cause of heart and vascular disease. At present, the global diabetes “epidemic” may be due to flavonoid starvation. Vegetarians are known to have a low incidence of diabetes and other degenerative diseases. It was common wisdom that the absence of diabetes could be related to the relatively low presence of sugar in vegetarian diets. Another explanation is that the diet is rich in flavonoids.

[0028] In addition to equivalents of the claimed elements, obvious substitutions now or in the future that are known to those of ordinary skill in the art are defined to be within the scope of the specified elements. It is therefore intended that the following claims be interpreted as including those particularly set forth and described above, conceptually equivalents, obvious substitutes, and essentially encompassing the essential concepts of the invention. It should be. Those having skill in the art will recognize that various adaptations and modifications of the preferred embodiment described above can be made without departing from the scope of the invention. The described embodiments have been presented by way of example only and are not to be construed as limiting the invention.

[Procedural Amendment] Submission of translation of Article 34 Amendment

[Submission date] May 2, 2001 (2001.5.2)

[Procedure amendment 1]

[Document name to be amended] Statement

[Correction target item name] Claims

[Correction method] Change

[Contents of correction]

[Claims]

[Procedure amendment 2]

[Document name to be amended] Statement

[Correction target item name] 0026

[Correction method] Change

[Contents of correction]

It is believed that the extract of Brichelia californica contains the flavonoids dihydroxychemferol, apigenin, luteolin, myricetin and quercetin. In addition, many other Brichelian species, although in varying proportions, contain these or similar flavonoids and should be effective in treating diabetes. Experiments with diabetic test animals (rats and mice) were performed. The extract of Brichelia was also effective in controlling blood glucose in these model systems. Also, administration of the synthesized Brichelia flavonoids also had the effect of lowering glucose levels. Luteolin was the most effective drug for treatment with a single flavonoid. However, the combination of luteolin with other flavonoids, especially dihydrochemferol and apigenin, has been shown to provide enhanced effects that can lower blood glucose maximally at lower total flavonoid doses. This effect is molar concentration of Ruteonin is seen the most expressed in the case of double least those combined it dihydro Chem Fellows Le and apigenin.

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Claims (10)

[Claims] 1. An anti-diabetic composition comprising an extract of a plant of the genus Brickellia. 2. An anti-diabetic composition comprising an extract of Brickellia californica. 3. An anti-diabetic composition comprising a flavonoid selected from the group consisting of luteolin, myricetin, dihydroxychemferol, apigenin, quercetin and mixtures thereof. 4. An anti-diabetic composition comprising a mixture of luteolin, dihydroxychemferol and apigenin. 5. The antidiabetic composition according to claim 4, wherein the molar concentration of luteolin is at least twice the combined concentration of dihydroxychemferol and apigenin. 6. A method for treating diabetes mellitus, comprising administering an extract of a plant of the genus Brixeria in an amount sufficient to cause a reduction in blood glucose. 7. A method for treating diabetes mellitus, comprising administering an extract of Brichelia californica in an amount sufficient to cause a reduction in blood glucose. 8. A method for treating diabetes mellitus comprising administering a flavonoid selected from the group consisting of luteolin, myricetin, dihydroxychemferol, apigenin, quercetin, and mixtures thereof, in an amount sufficient to cause a reduction in blood glucose. Method of treatment. 9. A method for treating diabetes mellitus comprising administering a mixture of luteolin, dihydroxychemferol and apigenin in an amount sufficient to cause a reduction in blood glucose. 10. The method of claim 9, wherein the molar concentration of luteolin is at least twice the combined concentration of dihydroxychemferol and apigenin.