JP2862676B2 - Prostate extract supplemented with zinc - Google Patents

Description

Description: FIELD OF THE INVENTION The present invention relates to compositions and formulations comprising prostate extracts supplemented with zinc.

United States Government Related Inventions This invention was made with United States Government support. The Department of Veterans Affairs
With assistance from facilities, equipment and materials. The United States Government has certain rights in the invention in the United States of America.

BACKGROUND OF THE INVENTION Diabetes is one of the most common metabolic disorders for humans. In fact, over one million Americans suffer from juvenile diabetes. This form of diabetes, also known as insulin-dependent diabetes mellitus or type I diabetes, develops suddenly approximately in childhood or adolescence. Type II or non-insulin dependent diabetes is characterized by a less sudden onset. Type II diabetes is about
The disease begins after the age of 40. Both types of diabetes impair the body's ability to access the blood sugar it uses as an energy source. Chronic high blood sugar levels gradually degrade various tissues and organs of the body.

Efforts have been made to understand the causes and consequences of diabetes. It is now believed that a potent autoimmune component is important in the pathogenesis of type I diabetes (see Science 25 : 1381 (1984); Immunology Toda).
y 5: 230 (1984)). The metabolic consequences of experimentally induced diabetes have also been investigated. For example, Schneier (Sc
have shown that the half-life of collagen, a key structural protein in the skin, was significantly reduced in diabetic rats (Diabetes 31 : 426 (1982)). It is well known that both types of diabetes affect a wide range of body tissues.

Despite the availability of insulin therapy, diabetes remains one of the serious illnesses and morbidity in the world with high mortality and morbidity. For example, the life expectancy of the average diabetic is reduced by as much as 50%. The prolonged illness of diabetes results in eye damage, often leading to blindness, cardiovascular problems, problems associated with wound healing, and other serious consequences.

SUMMARY OF THE INVENTION One aspect of the present invention is a composition comprising crystalline zinc chelated with an extract of animal prostate tissue.

Another aspect of the invention is a pharmaceutical composition that can be administered to a mammal. The pharmaceutical composition comprises crystalline zinc chelated with an extract of animal prostate tissue and at least one pharmaceutically acceptable excipient. The composition can be in the form of a tablet or capsule. Each tablet or capsule of the composition preferably contains about 20 to 150 milligrams of zinc.

Another aspect of the invention is a method of treating diabetes.
According to this method, a pharmaceutical composition comprising a zinc-chelated prostate extract is administered to a mammal suffering from diabetes in an amount sufficient to reduce the blood glucose level of the mammal.

Another aspect of the present invention is directed to a pharmaceutical composition comprising an extract of animal prostate tissue for use in treating diabetes. In a preferred embodiment, the pharmaceutical composition further comprises zinc.

Another aspect of the invention is the use of an organic extract of animal prostate tissue for use in the manufacture of a medicament for treating diabetes.

Another aspect of the invention is the use of a prostate extract for the treatment of diabetes. The extract is preferably supplemented with zinc. Also, the prostate extract can be used in the manufacture of a medicament for diabetes medicine. In one embodiment, the prostate tissue comprises a first group of molecules soluble in petroleum ether or hexane. A subset of molecules soluble in petroleum ether or hexane is present in prostate tissue. Prostate tissue also contains a second group of molecules that are soluble in ethyl acetate or chloroform. An extract of animal prostate tissue is obtained by firstly chopping the small pieces into small pieces and resuspending the finely chopped tissues in an aqueous solution. Thereafter, a first group of molecules can be extracted from the resuspended tissue using a first non-polar solvent that is less polar than petroleum ether and hexane. Thereafter, the first solvent containing the first group of molecules can be discarded. The second group of molecules can be extracted using a second solvent that is more polar than the first solvent. Finally, the desired prostate extract can be made by removing the second solvent.

BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 shows the zinc concentration in the organ tissues of normal and untreated normal rats treated with prostate extract. Open bars indicate rats that were not treated with the prostate extract, and shaded bars indicate rats that were treated with the prostate extract. In the figure, * indicates P <0.05, and ** indicates P <0.
Indicates 001.

FIG. 2 shows zinc concentrations in organ tissues of diabetic rats treated with the prostate extract and untreated diabetic rats.
Open bars indicate rats that were not treated with prostate extract, and hatched bars indicate rats that were treated with prostate extract. In the figure, * indicates P <0.05, and ** indicates P <0.
Indicates 001.

FIG. 3 is a diagram comparing zinc concentrations in organ tissues of normal rats and diabetic rats treated with the prostate extract and normal rats and diabetic rats not treated with the prostate extract.

NC refers to normal rats not treated with prostate extract.

 NP refers to normal rats treated with prostate extract.

DC refers to diabetic rats not treated with prostate extract.

 DP refers to diabetic rats treated with prostate extract.

In the figure, ** indicates P <0.00 when the value of the rat treated with the prostate extract was compared with that of the untreated rat.
1 and ## indicates P <0.001 when comparing the value of diabetic rats to that of normal rats.

DETAILED DESCRIPTION OF THE INVENTION I have found that compositions comprising zinc chelated with extracts of animal prostate tissue are useful for treating diabetes. The composition is also useful for treating other conditions that affect the metabolism of zinc and essential fatty acids.

For at least 60 years, the biomedical community has been aware of the fact that zinc metabolism changes as a result of diabetes. For example, Scott et al. Have shown that zinc is essential for the storage of insulin in human pancreatic B cells (J. Clin, Invest, 17 : 725 (1938)). Engelbart and Kief found that rapid stimulation of rat insulin secretion reduced the zinc content of pancreatic B cells (Virchows Arch. B4: 94).
−302 (1970)). It is also well known that zinc is involved in insulin storage and that the amount of insulin stored during zinc deficiency is lower than when zinc levels are normal. Therefore, an important link between zinc metabolism and diabetes has been clearly established in the chemical literature.

The experimental results presented in Example 1 confirmed that increased survival of experimental animals with diabetes was correlated with increased zinc levels due to diet. Based on these observations, I hypothesized that dietary supplements that facilitate zinc utilization would be useful for diabetics.

I have considered the prostaglandin as a therapeutic agent for diabetes that can improve the symptoms associated with diabetes. Son
ng) and Adohamu (Adham) at least some of the prostaglandins (PGE ₂₎ is a zinc - so it reveals previously that can act as a binding ligand (J.Nutr
ition 109: 2152 (1979)), and my research focuses on these substances. The authors of the report revealed that other prostaglandins (PGA ₂ and PGB ₂ ) had no effect on zinc transport. Thus, only a subset of the prostaglandins efficiently chelate zinc or otherwise affect zinc transport. In addition, Song and Mooradian consider that arachidonic acid, a prostaglandin precursor, plays a role in regulating zinc flux across the intestinal epithelium of diabetic rats (Life Sciences 42: 687 (198
8)). Arachidonic acid is one example of a prostaglandin precursor.

While it is preferred to alleviate the symptoms of diabetes by directly administering an appropriate prostaglandin, this method is impractical. Prostaglandins cannot be used as therapeutics because of their very short half-lives.

Since prostaglandins are synthesized from unsaturated fatty acid precursors, I have focused on this latter compound as a candidate for treating diabetes. Further, my research has focused on sources of unsaturated fatty acids that can be administered as a dietary supplement. In short, three different sources of unsaturated fatty acids were tested as treatments for diabetes. Comparison results of arachidonic acid, evening primrose oil, and a new extract of animal prostate tissue show that there is a clear difference in their efficacy as antidiabetic agents. In particular, animal prostate tissue extracts most effectively lowered blood glucose in laboratory rats and increased glucose concentrations in the cytosol. Most importantly, my research has demonstrated that encapsulated zinc-chelated prostate extract lowers blood glucose in diabetics. This result confirms the efficacy of the novel composition as a viable treatment for diabetes.

I. Crystalline Zinc Chelated with Prostate Extract One aspect of the present invention is zinc chelated with prostate extract. That is, it is a crystal in which negatively charged fatty acids present in the organic extract of prostate tissue are bound by positively charged zinc ions.

Preferably, the organic extract of prostate tissue contains unsaturated fatty acids including essential fatty acids. The essential fatty acid is preferably selected from the group consisting of prostaglandins and prostaglandin precursors. These fatty acids are obtained from the prostate of animals such as cows, sheep or goats. The extraction involves resuspending the prostate in a buffered aqueous solution, extracting saturated fatty acids with a highly non-polar solvent such as petroleum ether or hexane, extracting unsaturated fatty acids with a highly polar organic solvent such as ethyl acetate or chloroform, This can then be achieved by adding a zinc salt in an amount sufficient to chelate any fatty acids or amino acids present.

II. Pharmaceutical Composition Another aspect of the present invention relates to a pharmaceutical composition. According to the present invention, the pharmaceutical composition comprises (1) an extract of animal prostate tissue, (2) a zinc salt such as zinc sulfate, (3) a protein hydrolyzate, and (4) at least one
Contains two pharmaceutically acceptable excipients.

The protein hydrolyzate may be in amino acid form or incompletely hydrolyzed protein form such as proteose, peptone, or other partially hydrolyzed protein form such as albumin.

Pharmaceutical compositions prepared according to the present invention preferably contain essential amino acids, zinc sulfate, and protein hydrolyzate in a weight ratio of approximately 10: 1: 5. The pharmaceutical compositions can be formed into tablets or capsules by methods well known in the pharmaceutical art. Preferably, each tablet or capsule contains, in addition to one or more pharmaceutically acceptable excipients, about 200 mg of prostate extract, about 20 mg of zinc, and about 100 mg of protein hydrolyzate . Suitable excipients for tablets and capsules include inert diluents, such as calcium carbonate, sodium carbonate, or sodium bicarbonate, lactose, or calcium phosphate, or binders, such as starch, gelatin, or acacia. Or a lubricant such as magnesium stearate, stearic acid, or talc. Capsule coatings can use gelatin or soluble polymers, as is well known in the art. Tablets or capsules are suitable for oral administration.

The pharmaceutical compositions described herein are useful for treating diabetes, hypertension, weakness, and other diseases in which zinc or prostaglandin metabolism is impaired. In particular, diabetes can be treated by administering to a diabetic animal a zinc-chelated prostate extract of the present invention in an amount sufficient to reduce the blood glucose level of the mammal. A typical dose for a diabetic or hypertensive patient is about 80, expressed in terms of zinc.
Or about 150 mg of zinc. This dosage may be adjusted by persons of ordinary skill in the art depending on factors such as the weight, age, sex, and health of the patient, as well as the response to the particular dosage.

In the experiment described with reference to FIGS. 1 and 2, 2 hours before the rat was anesthetized and 2 hours before killing, 2 g of distilled water was injected by gastric intubation.
Were administered 200 mg of prostate extract and 200 μg of zinc. All values are SEM ± mean of 6 measurements. The composition of the prostate extract was determined to determine the zinc concentration in all test tissues of normal rats and the blood, heart, lung,
And that it had a sudden effect on the pancreas.

FIG. 3 shows a comparison of zinc concentration in organ tissues between normal rats and diabetic rats treated with the prostate extract and normal rats and diabetic rats not treated with the prostate extract, based on the data shown in FIGS. It was done. Normal and diabetic rats treated with prostate extract;
The average value of the zinc concentration in the tissue was compared between the two rats that were not treated. There is a significant difference between normal and diabetic rats, and it can be seen that prostate extract had a significant effect on normal and diabetic rats.

The experiments described in Example 1 below show that there is a fundamental relationship between the level of dietary zinc consumption and longevity in diabetic rats. In this method, laboratory rats were given diabetes by injecting chemicals well known in the art. Thereafter, the diabetic rats were fed a diet that contained insufficient to excessive amounts of zinc. The results of this simple experiment demonstrated that dietary zinc affected the longevity of diabetic rats.

Example 1 Effect of Dietary Zinc on the Survival of Diabetic Rats 51 rats were divided into three groups of 17 rats each. All rats were given diabetes by injection of streptozotocin (50 mg / kg). One week later, 3
Two groups of rats were fed a diet containing a defined amount of zinc. The first group has a diet depleted of zinc (1 μg Zn / g), the second group has a diet full of zinc (37.5 μg Zn / g), and the third group has an excess of zinc. Food (1 mg Zn / g). After 25 days, the number of surviving rats was counted. The results shown in Table 1 summarize the survival of the test animals in this experiment.

Example 2 describes the method used to separate unsaturated fatty acids from animal prostate tissue.

Prostate tissue contains zinc, prostaglandins, essential amino acids, citrate, testosterone and proteins.
However, there is no evidence that organic extracts of prostate tissue are limited to these substances.

Example 2 Formulation of crystalline zinc chelated with prostate extract Bovine prostate from a slaughterhouse was minced into small pieces and frozen at -70 °. More than 10 times the sliced tissue (W /
V) in 6.0 mM Tris-HCl buffer, pH 8.0, and homogenizer of VIRTIS-45 (Virtis Co., Cardne
(New York) at 4 ° C. for 20 minutes at 3000 × g. The top of the fat was physically removed and the supernatant was kept at 37 ° C. for 1 hour. Saturated fatty acids were extracted with petroleum ether. The remaining aqueous solution containing unsaturated fatty acids was adjusted to pH 3.0 using 0.2 N HCl, and the mixture of unsaturated fatty acids containing prostaglandin was mixed with isopropanol or chloroform each time (1). : 1 v / v) ethyl acetate and extracted two or three times. Thereafter, the ethyl acetate or chloroform extract was mixed. Freeze-drying the unsaturated fatty acid solution, or
Alternatively, it was dried by evaporation under reduced pressure. Prostate extracts such as oils resulting from such operations contained unsaturated fatty acids. Formulations of unsaturated fatty acids chelated with zinc were combined with 20 mg of zinc chloride and Sigma (St, L
ouis, MO), prepared with 200 mg of prostate extract mixed with 100 mg of protein hydrolyzate.

Example 3 describes the method used to evaluate the effects of arachidonic acid, evening primrose oil and prostate extract on the symptoms of diabetic rats. Three samples of unsaturated fatty acids were tested for their relative ability to improve symptoms in diabetic rats. Arachidonic acid and evening primrose oil were the two substances used in such a comparison. Evening primrose oil is known to be rich in both linoleic acid and linolenic acid.
Alternatively, I prepared and tested an organic extract of animal tissue as a sample of unsaturated fatty acids that could positively affect diabetic symptoms in rats. All substances were administered as a dietary supplement with dissolved zinc salts. My findings show that prostate extracts most beneficially affect the symptoms of diabetic rats.

Example 3 Investigation of the Effect of Unsaturated Fatty Acids on the Symptoms of Diabetic Rats First, streptozotocin was injected to diabetic rats. After that, these diabetic rats were given 3
Drinking water containing one of the three unsaturated fatty acid formulations was administered. Drinking water contained zinc sulfate dissolved with arachidonic acid, evening primrose oil or prostate extract prepared according to the method of Example 2. The zinc sulphate solution was diluted to a final concentration of 10 mg / l of zinc. Arachidonic acid and prostate extract were diluted to a final concentration of 100 mg / l each. Evening primrose oil at a final concentration of 500 mg / l
It was diluted so that Used in this manner, the concentrations of zinc and prostate extract were chosen to mimic proportionately to a single dose given to an adult. In the concentration calculations, it was assumed that a diabetic patient weighing 60 kg consumed 600-1000 mg of prostate extract and 60-100 mg of zinc daily. This is 10 mg of prostate extract and 1 mg of zinc per kg of body weight
Reach The estimated daily water consumption of normal rats was 40 ml. 18 after injection of streptozotocin
GLUCOMETER ELITE: Mil
es Co.) was used to record blood glucose levels in all test animals. Table 2 shows the results of these glucose measurements.

The results shown in Table 2 confirm that animals with diabetes have significantly higher blood glucose levels than normal rats.

Administering zinc alone to diabetic rats had no effect on blood glucose levels. Administering zinc with evening primrose oil in food also had no effect on lowering blood glucose levels in diabetic animals. Combined treatment with zinc and arachidonic acid reduced blood glucose levels to normal levels. Surprisingly, the decrease in blood glucose that appeared after administering zinc and prostate extract was more significant than any other treatment.

It was unexpected that in contrast to prostate extract effectively lowering blood glucose levels in diabetic animals, evening primrose oil and arachidonic acid would be ineffective or semi-effective. Was. Importantly, this work has demonstrated that not all unsaturated fatty acid samples are equivalent in their ability to ameliorate diabetes symptoms.

To independently evaluate the second indicator of diabetes symptoms,
Cytosolic glucose levels were measured in various tissues of diabetic rats fed various dietary supplements. Example 4 shows the results of such measurements performed using rats treated as described in Example 3.

(Example 4) Cytosolic glucose concentration in various tissues of diabetic rats Immediately after sacrificing the experimental animal obtained from Example 3 by injecting nembutal (50 mg / kg), various tissues were separated. Stored at 70 ° C. Each tissue is disrupted with a polytron homogenizer, and then
Centrifuged at 00xg for 20 minutes only. Supernatants were either freeze-interpolated or used immediately to determine glucose and protein concentrations. Glucose concentration (Yellow Springs
Measurements were made using a Model A23 glucose analyzer from Instruments (Yellow Springs Ohio). Lowr protein concentration
y et al. (J. Biol. Chem. 193 : 265 (1951)). The values obtained for the glucose readings were divided by the protein concentration readings to normalize the measurements. This normalization was necessary for variable mass and control of protein yields that differentiated batches of different tissues. Table 3 shows the results.

The results shown in Table 3 showed that there was a correlation between food supplementation with prostate extract and increased glucose uptake at the cellular level. Diabetic tissue is typically
Although the level of free glucose was low, my findings showed that this level was dramatically increased in test animals administered a combination of prostate extract and zinc. Administration of arachidonic acid or evening primrose oil in food in combination with zinc had no effect on cytosolic glucose concentration. The next developmental stage of the present invention was to extend the use of zinc-chelated prostate extracts beyond the treatment of laboratory animals.

The effect of prostate extract on relieving symptoms in diabetics was investigated. Example 5 shows the change in blood glucose levels of diabetics recorded after supplementing meals with capsules containing prostate extract chelated with zinc.

Example 5 Effect of Food Administration of Prostate Extract on Blood Glucose Levels of Diabetic Patients Effect of recruiting 16 diabetic patients as volunteers and supplementing with zinc-chelated prostate extract with food Was tested. GLUCOMETER ELIT
E) (Miles CO.) Was used to measure the initial blood glucose readings of all patients. Thereafter, all 16 volunteers took the chelated zinc-encapsulated prostate extract two to four times daily for one to three months. Each capsule is 200m
g of prostate extract and 20 mg of zinc and gelatin. These capsules were prepared according to the method described in Example 2 using a prostate extract chelated with zinc.
nner Pharmacap Co (Chatsworth, CA). At the end of the study period, all patients' blood glucose was measured. Table 4 summarizes the results of all blood glucose measurements in this experiment.

Dietary administration of zinc-chelated prostate extract and the rate of glucose removal from diabetic patients before and after curing were also measured. In this experiment, the rate at which a well-defined amount of glucose was removed from the bloodstream of an individual diabetic patient was measured. The rate of glucose removal is clearly higher in non-diabetic people, but lower in diabetic patients.
This low glucose removal rate is due to the reduced insulin activity characteristic of diabetes. The increase in glucose removal rate that appears after administration of a zinc-chelated prostate extract is an indication of a therapeutic effect.

The advantage of using the glucose removal rate as an indicator of efficacy is that this test method is not affected by food, drink or temporary changes in exercise, but blood glucose levels are. For this reason, a comparison of the blood glucose removal rates before and after administering a zinc-chelated prostate extract with food and drink and curing is considered to be the most accurate measure of the positive effect of this regimen. .

Example 6 describes the method used to determine the effects of food and drink administration of zinc-chelated prostate extract on glucose removal rates in volunteer diabetic patients.

(Example 6) Effect of zinc-chelated prostate extract on glucose removal rate in diabetic disease First, a medical center (Sepulveda VAMedical Cente)
In r), four volunteer diabetic patients were identified. After stabilizing their carbohydrate intake to a level of 150-200 g per day for 3 days, a solution 225 containing 75 g glucose was
After drinking ml, the glucose removal rate was measured. Blood glucose was measured at 0, 30, 60, 90, 150 and 180 minutes after drinking the glucose solution. For blood glucose measurement, ONE-TOUCH GLUCOMETER (Lifescan,
Inc., Milpitas, CA). Regarding the removal rate, the rate at which the glucose concentration decreased after reaching the maximum concentration was measured.

After determining the initial blood glucose removal rate, three randomly selected volunteers were given capsules of zinc-chelated prostate extract four times daily for three months. The composition of the capsule used in this method is the same as described in Example 5. The fourth candidate took a placebo. At the end of the three month run, the blood glucose removal rates of all volunteers were measured again. Table 5 shows the results of this method.

The results presented in Table 5 showed that the glucose removal rates of all diabetic patients after taking the prostate extract chelated with zinc for 3 months were significantly improved. Patients taking placebo did not show any such improvement. Because patients on placebo had less diabetes, their initial glucose removal rate was closer to normal than any other volunteer.

Interestingly, patients taking placebo in this study exhibited a reduced rate of glucose removal during the course of the experiment. This is indicative of the normal tendency of such patients, as the progressively heavier diabetes slows the glucose removal rate.

The findings presented here indicated that diet supplemented with zinc-chelated prostate extract ameliorated the symptoms associated with diabetes. The mechanism by which prostate extract improves symptoms in diabetic animals and humans is not clearly understood, but may be due to the strong effect of prostate extract on increased intestinal zinc absorption . Regardless of the mechanism, I find that supplementing food with dietary supplements of prostate extract can reduce the risk of diabetic symptoms, compared to regimens that include administering zinc alone or in combination with other unsaturated fatty acids in food and drink. It proved that even better results could be obtained in connection with the improvement.

The present invention provides compositions of convenient sources of both zinc and essential fatty acids. The composition can be used as a dietary supplement for the treatment of diabetes, or other conditions. Because the metabolism of zinc and the metabolism of essential fatty acids, including prostaglandin precursors, are interconnected, using such compositions is more important for treating diabetes than using zinc or fatty acids alone. More effective.

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Claims (10)

(57) [Claims] A composition for the treatment of diabetes comprising crystalline zinc chelated with an extract of animal prostate tissue. 2. A pharmaceutical composition in a form that can be administered to an animal, comprising: (a) the composition of claim 1; and (b) at least one pharmaceutically acceptable excipient. 3. The composition according to claim 1, wherein the composition is in the form of a tablet.
A pharmaceutical composition as described. 4. A pharmaceutical composition according to claim 3, wherein each tablet contains approximately 20 milligrams of zinc. 5. The pharmaceutical composition according to claim 2, which is in the form of a capsule. 6. The pharmaceutical composition according to claim 2, wherein said composition contains about 20 milligrams to about 150 milligrams of zinc. 7. The tissue contains a first group of molecules soluble in petroleum ether or hexane and a second group of molecules soluble in ethyl acetate or chloroform, and the extract is prepared by a method comprising the following steps: 2. The composition according to claim 1, which is obtained: (a) a step of finely chopping the tissue; (b) a step of resuspending the finely chopped tissue in an aqueous solution; Extracting the first group of molecules from the suspended tissue into a first non-polar solvent that is less polar than petroleum ether and hexane; (d) discarding the first solvent containing the first group of molecules; (e) A) extracting the second group of molecules into a second solvent that is more polar than the first solvent; and f) removing the second solvent to obtain the extract. 8. A composition comprising an extract of animal prostate tissue and zinc for use in treating diabetes. 9. The composition according to claim 8, which is used for producing a medicament for treating diabetes. 10. The composition of claim 9, wherein said organic extract of animal prostate tissue is zinc-chelated.