MXPA99007232A - Arginine silicate inositol complex and use thereof - Google Patents

Abstract

An arginine silicate-containing complex and its use in the prevention and treatment of atherosclerosis, as a dietary supplement and for promotion of structural integrity of bones and joints. The arginine silicate complex is synthesized by combining arginine, potassium silicate and inositol. The complex is administered orally three times per day in an amount ranging from about 250 mg to about 2,500 mg as a preventative or therapeutic agent.

Description

COMPLEX OF ARG1N1NA-SILICATO-INOSITOL AND ITS USE Field of the Invention The present invention relates to an arginine silicate complex and its use in the prevention and treatment of atherosclerosis, as a dietary supplement and for the promotion of the structural integrity of bones and cartilages. BACKGROUND OF THE INVENTION Atherosclerotic disease is a complex and chronic disease, involving the gradual accumulation of lipids, collagen, elastic fibers and proteoglycans in the arterial wall. Current methods of treating it include a low-fat diet, exercise and various drugs that lower cholesterol. Although these methods may signify the delay of the progression of atherosclerosis, they are not entirely satisfactory. Heparin sulfate proteoglycans (HSPGs) produced by the vascular endothelium are thought to retard the migration, multiplication and phenotypic transition of smooth muscle vascular cells, which play a central role in the atherogenic process, and in the maintenance of a luminal anticoagulant surface by the binding and activation of antithrombin III (Clowes et al., Nature, 265: 625-626, 1977, Guyton et al., Circ Res., 46: 625-634, 1980; Edelman et al. ., Proc. Nati, Acad. Sci. USA., 87: 3773-3777, 1990). It has been shown that various silicon compounds, administered orally or parenterally, inhibit cholesterol-induced intimal hyperplasia (atherosclerosis) in rabbits (Loeper et al., Athersclerosis, 33: 397-408, 1979; Loeper et al., in Biochemistry of Silicon and Related Problems, Plenum Press, New York, 1918, pages 281-296, Garson et al., ". Pharm. Sci., 601113-1127, 1971.) Injection or ingestion of available silicon compounds nutritionally (eg, monomethyltrisilanol, lysine silicate, sodium silicate) prevented the characteristic intimal thickening and fragmentation of the arterial elastic fibers observed in atherosclerosis, and several epidemiological studies report that increased dietary admissions of silicon are associated with reduced risk of coronary heart disease in humans (Schwrz et al., Lancet, i: 454-457, 1977; Schwarz et al., Lancet, i: 538-539; Bassler, Bri. Med. J., 1: 919, 1 978; Parr, Lancet i: 1087, 1980). Studies in young growing rats and chickens show that severe dietary deficiency of silicon results in abnormal structures of bones and joints, apparently due to the abnormal production of collagen and mucopolysaccharides (Carlisle, J. "Nutr. 106: 478-484 , 1976; Carlisle, J. Nutr 110: 1046-1055, 1980) .Silicon promotes the synthesis of collagen and mucopolysaccharides in vi tro (Carlisle et al., Fed. Proc. 37: 404, 1978; Carlisie et al. ., Fed. Proc. 39: 787, 1980.) The biochemical methods by which silicon achieves this effect are unknown.Silicone has been shown to increase bone mineral density.When administering an organosilicon compound (monomethyltrisilanol) to a postmenopausal woman, by injection at a dose of 50 mg twice weekly, the femoral density increases significantly by an average of 4.7% in 14 months of administration (Eisinger et al., Mgnesium Res. 6247-249, 1993). ovariotomized s, oral orthosilicic acid showed a change in bone and an increased rate of bone formation (Hott et al., Calcif. Tissue Int. 53: 174-179, 1993). Bone and cartilage are dynamic tissues in both juvenile and adult animals. In bone, osteoclasts solubilize the hydroxyapatite bone matrix and degrade collagen, while osteoblasts concurrently reconstruct bone through the synthesis of collagen and hydroxyapatite deposition. Similarly, the chondrocytes in the cartilage simultaneously degrade the collagen and the proteoglycan matrix and resynthesise them. The impact of silicone on bone and the formation of cartilage in adult animals is essentially unknown. However, it is highly probable that the role of silicon in the metabolism of bone and cartilage is limited to juvenile animals. The nutritional role of silicon is to support the adequate synthesis of mucopolysaccharides, proteoglycans and collagen (Schwarz et al., Nature 239: 333-334, 1972; Carlisle, Science, 178: 619-621, 1972; Carlisle, J. Nutr. , 106: 478-484, 1976, Schwarz, in Biochemistry of Silicon and Related Problems, Plenum Press, New York, 1978, pages 207-230). Optimal silicon nutrition can promote the production of protective HSPGs by endothelial cells. Arginine, an essential amino acid, is the biosynthetic precursor for nitric oxide (NO) produced by the vascular endothelium (Moneada, New Engl. J. Med., 329: 2002-20012, 1993). This NO exerts vasodilatory, antiateroesclerotic and antithrombotic effects, and deficient endothelial production in this NO, does not play a prominent pathogenic role in atherosclerosis, hypertension and diabetes (Calver et al., J. Hypertension, 10: 1025-1031, 1992; Cooke et al., Arterioscler, Thromb., 14-653-655, 1994; Rubanyi, in Cardiovascular Significance of Endo thelum -Der i ved Vasoactive Factors, Futura Publishing Co., Inc., New York, 1991, pages xi -xix). In some, but not all, clinical studies, parenteral or oral administration of arginine has increased the synthesis of vascular NO (Drexler et al., Lancet, 338.1546-1559m 1991). In animal models of hypertension, arginine supplementation has moderated the increase in blood pressure (Chen et al., "Clin. Invest., 88: 1559-1567, 1991; Laurant et al., Clin.

Exp. Hyperten. , 27: 1009-1024, 1995). Thus, under at least some circumstances, the availability of arginine may be a limitation of the regime for the production of NO. A recently published clinical study indicates that oral arginine may increase endothelium-dependent relaxation in young hypercholesterolemic individuals (Creager et al., J., Clin.Res 90: 1248-1253, Clarkson et al., J. Clin. ., 97: 1989-1994, 1996) which is indicative of the increased efficiency of vascular NO production.There is a constant need for therapeutic / prophylactic agents, capable of preventing or slowing the progression of atherosclerosis and promoting bone formation. and cartilage The present invention addresses these needs.

SUMMARY OF THE INVENTION One embodiment of the present invention relates to a method for obtaining an arginine-silicate-inositol complex, which comprises the steps of: (a) combining the arginine, a silicate salt and the inositol, to form a suspension; (b) heating the suspension to promote the formation of a gel; (c) allow the gel to crystallize; (d) mixing the crystals, formed in step (c), with an alcohol, to promote crystallization; and (e) collecting the crystals formed in step (d). Preferably, the silicate salt is potassium silicate. Advantageously, the heating is carried out at about 95 ° C. The method can also include repeating the stage (d), before collecting the crystals. In one aspect of this preferred embodiment, the crystals are collected by filtration. Preferably, the alcohol that promotes crystallization is ethanol. The present invention also provides an arginine-silicate-inositol complex, by the method described above. Another embodiment of the invention relates to a method for preventing or inhibiting atherosclerosis in a mammal, preferably a human, comprising the step of administering to the mammal an effective amount that prevents or inhibits the atherosclerosis of the arginine-silicon-inositol complex, previously described. Preferably the administration step is the parenteral or oral route. Advantageously, the effective amount is between about 250 mg and 2,050 mg; more advantageously, the effective amount is between about 500 and 1,000 mg. For the average 70-kg man, this equals a dose between 3.6 and 14 mg / kg (250-2,500 mg) and between approximately 7.1 and 14 mg / kg (500-1,000 mg), respectively. Yet another embodiment of the invention relates to a method for supplementing the diet of arginine, which comprises administering an individual to the complex described above. The present invention also provides an arginine-silicate-inositol complex, wherein the ratio of arginine to silicate The inositol is approximately 3: 3: 1. Another embodiment of the present invention relates to the use of the arginine-silicon-inositol complex, described above, to supplement dietary arginine.

The present invention also provides in use the arginine-silicon-inositol complex, described above, for the prevention or inhibition of atherosclerosis. Another embodiment of the invention relates to a method for preventing bone demineralization or cartilage degradation in an individual in need, this method comprises administering to the individual an amount that inhibits effective bone demineralization or cartilage degradation of the complex. of arginine silicate, described above. Yet another embodiment of the invention relates to the use of the arginine silicate complex, described above, to prevent the demineralization of bones or the degradation of cartilages in an individual. Preferably, the route of administration is parenteral or oral. The present invention also provides a method for treating bone or cartilage disorders in an individual in need thereof, this method comprising administering to the individual an effective amount of the arginine silicate complex., described above. In one aspect of this preferred embodiment, this bone disorder is osetoporosis, osteogenesis imperfecta or bone fractures. Preferably, the cartilage disorder is osteoarthritis, inflammatory arthritis, a torn tendon or a torn ligament. Advantageously, the administration is parenteral or oral. Yet another embodiment of the invention is the use of the arginine silicate complex, described above, for treating bone or cartilage disorders in an individual. Another embodiment of the invention is a pharmaceutical formulation that opposes the demineralization of bones and that inhibits cartilage degradation, which comprises a therapeutically effective amount of the arginine silicate complex, described above; and its pharmaceutically acceptable carrier or diluent. The present invention also provides a method for treating a mammal to alleviate the pathological effects of osteoporosis, osteogenesis imperfecta, bone fractures, osteoarthritis, inflammatory arthritis and other bone and cartilage disorders, wherein the method comprises administering to this mammal the complex of arginine silicate, described above, wherein the complex is administered to the mammal in an amount sufficient to oppose bone demineralization and to inhibit cartilage degradation.

Detailed Description of the Preferred Modes The present invention provides a complex of * arginine silicate, produced by the combination of arginine, a silicate salt and inositol, a method for its synthesis and its use as a nutritional supplement, for the prevention and treatment of atherosclerosis and to promote the structural integrity of the bone and cartilage. Although the product described here contains arginine, silicate and inositol, it is named through the specification as the "arginine silicate". The arginine silicate is synthesized by the reaction of arginine (free base), potassium silicate and inositol, as described in Example 1. The resulting complex is completely soluble and supplies the silicate in a bioavailable form, which will have a good nutritional availability. The silicates are typically insoluble in aqueous solutions. However, the use of inositol in the synthesis of the complex containing the arginine silicate, makes the complex soluble in aqueous solutions. In contrast, the arginine silicate synthesized in the absence of inositol was insoluble in aqueous solutions. This effect of unexpected solubilization of inositol is of paramount importance in the use of the complex as a bioavailable source of arginine and silicate. Inositol facilitates the solubilization of arginine silicate, increasing the hydrogen bond between arginine and silicic acid. Although other polyhydroxy compounds, including, but not limited to, mannitol and sorbitol, can also be used, inositol is preferred. The bioavailability of the silicate was confirmed as described in Example 3. In a preferred embodiment, the molar ratio of the combination of arginine to the silicate is about 1: 1 and the ratio of inositol to arginine and silicate is about 1. :3. Although potassium silicate is used as a reagent, the use of other silicate salts, which include sodium silicate and magnesium silicate, is also within the scope of the invention. The mixture that results from the combination of inositol, silicate salt and arginine, is a highly viscous suspension, which is clarified by heating.

In a preferred embodiment, the suspension is heated between about 80 and 100 ° C, more preferably at about 95 ° C, until clarification is observed. At this time, heating and stirring are discontinued and gel formation is initiated. The crystallization of the arginine silicate complex occurs as the formation of the gel progresses. The resulting crystal volume is dispersed and mixed with an alcohol for about 30 minutes, to effect the most complete crystallization and the recovery of a purer product. The heavy metal content of the final product was less than 5 ppm, which is considered undetectable. The iron level was also "very low (10 ppm)." These findings indicate that the product is virtually free of such contaminants.Although the use of ethane for the crystallization of the arginine silicate complex is preferred, the use of Other alcohols Optionally, a second stage of alcohol crystallization can be carried out.The final product, a complex containing arginine, silicate and inositol, is collected by filtration, washing and drying.The arginine silicate can be used both as a source of arginine as an essential amino acid and as a source of silicate, both of which exert anti-atherosclerotic effects.Oral administration of this compound delivers arginine and silicate at appropriate sites of action.The arginine silicate is useful as a therapeutic agent and preventive for atherosclerosis and can also be provided as a dietary supplement to maintain an antiatherogenic state. Thus, the administration of arginine silicate has a prophylactic as well as a therapeutic effect. Arginine silicate is highly soluble in water and provides good nutritional availability of both arginine and silicate. further, to supply the silicate, the arginine silicate complex is also a good dietary supplement for the essential amino acid, arginine. The arginine silicate complex of the invention promotes the formation of bone and cartilage in a mammal that needs it, particularly in humans. The bioavailable nutritional silicon, in the form of the arginine silicate complex, described herein, also increases bone density and prevents demineralization thereof. In a preferred embodiment, the complex is administered prophylactically, to prevent bone demineralization and cartilage degradation. A preferred use of the complex is the prevention and treatment of osteoporosis, which results from bone demineralization, in postmenopausal women. The complex is used to prevent or treat any bone demineralization disorder, which includes osteoporosis and osteogenesis imperfecta. The arginine silicate complex is also used as an aid in the treatment of bone fractures. For example, an individual with a bone fracture is treated by shingling in combination with oral administration of the arginine silicate complex of the invention, to promote faster recovery of the fracture. This decreases the time that an individual must use the cast, in situations where this applies. The arginine silicate complex can also be used to treat "fresh adhesion" fractures, in which no actual separation of the bone has occurred. In yet another preferred embodiment, the arginine silicate complex is used to treat or prevent osteoarthritis and inflammatory arthritis. In yet another preferred embodiment, the arginine silicate complex is administered to an individual with torn cartilages or tendons, either alone or after surgery, to repair damaged areas. Promoting the formation of cartilage, the arginine silicate complex decreases the recovery time after surgery. The compounds of the invention may be administered parenterally, orally, intravenously, intraarterially, intramuscularly or in any other systemic form, in appropriate dosage units, as desired. The term "parenteral", used herein, includes the techniques of subcutaneous, intravenous, intraarterial injection or infusion, without limitation. However, oral administration is preferred. For oral administration, the compounds may be provided as a tablet, aqueous or oral suspension, dispensable powders or granules, hard or soft capsule emulsion, syrup or elixir. The compositions intended for oral use can be prepared according to any method known in the art, for the manufacture of pharmaceutical compositions and such compositions can contain one or more of the following agents: sweeteners, flavoring agents, color people, preservatives , solubilizers, moisturizing agents, stabilizers, colorants, antioxidants, coating agents and diluents. The sweetening agents and flavoring agents will increase the acceptability of the preparation. Tablets containing the arginine silicate in admixture with non-toxic, pharmaceutically acceptable, soluble excipients for the manufacture of tablets are acceptable. Such excipients include inert diluents, such as calcium carbonate, sodium carbonate, lactose, calcium phosphate or sodium phosphate.; granulating and disintegrating agents, such as corn starch or alginic acid; binding agents, such as starch, gelatin or acacia; and lubricating agents, such as magnesium stearate, stearic acid or talc. The tablets may be uncoated or they may be coated by known techniques, to retard disintegration and absorption in the gastrointestinal tract and thus provide a sustained action for a longer period of time. For example, a delay material, such as glyceryl monostearate or glyceryl distearate, can be used alone or with a wax. Formulations for oral use may also be present as gelatin capsules, in which the active ingredient is mixed with an inert solid diluent, for example calcium carbonate, calcium phosphate or kaolin, or as soft gelatin capsules, wherein The active ingredient is mixed with water or an oil medium, such as peanut oil, liquid paraffin or olive oil. Aqueous suspensions may contain the compounds of the invention in admixture with excipients suitable for the manufacture of aqueous suspensions. Such excipients include suspending agents, dispersing or hydrating agents, one or more preservatives, one or more color agents, one or more flavoring agents and one or more sweetening agents, such as sucrose or saccharin. Oil suspensions can be formulated by suspending the active ingredient in a vegetable oil, such as arachis oil, olive oil, sesame oil or coconut oil, or in a mineral oil, such as liquid paraffin. The oil suspension may contain a thickening agent, such as beeswax, hard paraffin or cetyl alcohol. Sweetening agents, such as those noted above, and flavoring agents, may be added to provide an acceptable oral preparation. These compositions can be preserved by an added antioxidant, such as ascorbic acid. The powders and dispersible granules of the invention, suitable for the preparation of an aqueous suspension, by the addition of water, supply the active ingredient in admixture with a dispersing or moisturizing agent, a suspending agent and one or more preservatives. Additional excipients, for example sweetening agents, of taste and color, may also be present. The syrups and elixirs can be formulated with sweetening agents, such as glycerol, sorbitol or sucrose. Such formulations may also include an emollient, a preservative, a flavoring or coloring agent. To assist in formulating the compositions of the present invention, one may refer to the Remington 's Pharmaceutical Sciences, 15th Edition, Mack Publishing CO., Easton, PA. The arginine silicate preparations, for parenteral administration, may be in the form of a sterile injectable preparation, such as a sterile, injectable aqueous or oleaginous suspension. This suspension can be formulated according to methods well known in the art, using suitable dispersing or hydrating agents and suspending agents. The sterile injectable preparation can also be a sterile injectable solution or suspension in a non-toxic, parenterally acceptable diluent or solvent, such as a solution in 1,3-butanediol. Suitable diluents include, for example, water, Ringer's solution and an isotonic sodium chloride solution. In addition, fixed sterile oils can be conventionally employed as a solvent or suspending medium. For this purpose, any fixed soft oil may be employed, which include the synthetic mono- or diglycerides. In addition, fatty acids, such as oleic acid, can similarly be used in the preparation of injectable formulations. Optionally, the pharmaceutical compositions of the invention may comprise the arginine silicate complex combined with one or more compounds, which exhibit a different activity, for example, an antibiotic or other pharmacologically active material. The amount of the arginine silicate that can be combined with the carrier material, to produce a single dose form, will vary depending on the host treated and the particular form of administration. In a preferred embodiment, as a preventive or therapeutic agent for atherosclerosis or disorders of bone and cartilage, the arginine silicate is administered three times a day, in an amount ranging from 250 to 2,500 mg. In a particularly preferred embodiment, the compounds are administered three times a day in an amount ranging from about 500 to 1,000 mg. It is also considered that the compounds can be administered once or twice a day, rather than three times, depending on the severity of the atherosclerotic lesion. The arginine silicate was synthesized as described in the following example.

Example 1 Preparation of Arginine Silicate Arginine (3.9 g, 21.8 mmol) was added to a vigorously stirred solution of inositol (1.25 g, 6.9 mmol) in potassium silicate (5 mL, 29.8 ° Be, 8.3% K20 (0.52 g, 5.5 mmol), 20.8% Si02 (1.3 g, 21.8 mmol), which results in a highly viscous suspension. The suspension was heated to 95 ° C. The heating and stirring were discontinued when the mixture became clear and began to form a gel. The mixture was left overnight at room temperature and allowed to crystallize. The resulting volume of crystals was dispersed, mixed with ethanol (5 ml) and left for 30 minutes. This procedure was repeated with another 5 ml of ethanol in the resulting crystals and left overnight to complete the crystallization. The final product of arginine silicate was collected by filtration, washed with ethanol and dried in vacuo. The quantity of the product was 7.7 g, obtained as a hydrate (111% of the total mass of the reagents used) An analytical sample, which was kept under vacuum at 90 ° C for 1 hour, lost 11.5% of its mass to the water removal The elemental analysis indicated: 25.13% of C, 6.24% of H, 14.11% of N, 8.25% of Si (17.68% of Si02) .The content of potassium (5.4%) was determined using an equipment (HACH Co., Loveland CO, Catalog No. 234394) based on the well-known tetraphenylborate method, these results are in accordance with the calculated content of elements in the arginine silicate product, eg 2 Kinetics of the silicate product. of arginine The kinetics studies in the aqueous solution of the arginine silicate product indicated the formation of a non-dissociable arginine silicate complex, as a function of the concentration used.The measurement of the ratio of the dissociated to non-dissociated forms of the arginine silicate is performed using the HACH equipment (Catalog No. 24296-00) in which the absorbance at 452 nm is a function of the concentration of the silicomolybdate formed under acidic conditions and expressed as% silica (Si0?). An aqueous solution of the arginine silicate product (10 g / 1) was diluted at the appropriate time to 0.5 g / 1 and the silica content was measured using the JHACH method. The level of silica at time 0 was 17.5%; in 1 hour it was 11.8%; in 2 hours it was 10.8%; and in 24 hours it was 9.2%. In an aqueous solution of 0.5 g / 1 of arginine silicate, the silica level was 17.5% and was stable after 24 hours, confirming the solubility of the product.

Example 3 Bioavailability of arginine silicate A solution (8 g / 1) of arginine silicate was prepared and, after donating a basic 24-hour urine line, a human volunteer consumed three portions of a cup daily for three days . On the third day, he again obtained a 24-hour urine. The silicon test revealed that the production of silicon in the urine had increased more than ten times from the baseline. The amount of silicon in the urine of the third day corresponded to approximately 25% of the silicon ingested daily from the solution of arginine silicate. This demonstrates the good bioavailability of silicon in the solubilized arginine silicate. While particular embodiments of the invention have been described in detail, it will be apparent to those skilled in the art that these embodiments are exemplary, rather than limiting, and the true scope of the invention is that defined in the appended claims.

Claims (25)

1. CLAIMS 1. A method to obtain an arginine-silicate-inositol complex, which comprises the steps of: a. combine arginine, a silicate salt and inositol, to form a suspension; b. heating the suspension to promote gel formation; c. allow the gel to crystallize; d. mixing the crystals, formed in step (c), with an alcohol, to promote crystallization; and e. Collect the crystals formed in step (d).
2. 2. The method of claim 1, wherein the silicate salt is potassium silicate.
3. 3. The method of claim 1, wherein the heating is performed at about 95 ° C.
4. 4. The method of claim 1, further comprising repeating step (d), before collecting the crystals.
5. 5. The method of claim 1, wherein the crystals are collected by filtration.
6. 6. The method of claim 1, wherein the alcohol, in step (d), is ethanol.
7. 7. An arginine-silicate-inositol complex, formed by the method of claim 1.
8. 8. A method for preventing or inhibiting atherosclerosis in an individual, comprising the step of administering to this individual an effective amount, which prevents or inhibits atherosclerosis, from the arginine-silicon-inositol complex of claim 7.
9. 9. The method of claim 8, wherein the step of administration is parenteral or oral.
10. 10. The method of claim 8, wherein the effective amount is between about 250 and 2,500 mg.
11. 11. The method of claim 10, wherein the effective amount is between about 500 and 1,000 mg.
12. 12. A method for supplementing dietary arginine, which comprises administering to an individual the complex of claim 7.
13. 13. An arginine-silicate-inositol complex, in which the ratio of arginine to silicate to inositol is approximately 3: 3: 1.
14. 14. The use of the complex of claim 7, to supplement the dietary arginine.
15. 15. The use of the arginine-silicon-inositol complex of claim 7, for the prevention or inhibition of atherosclerosis.
16. 16. A method for preventing bone demineralization or cartilage degradation, in an individual in need thereof, this method comprises administering to the individual an effective amount that inhibits bone demineralization or cartilage degradation, of the complex of claim 7.
17. 17. The use of the claim complex 7, to prevent bone demineralization or cartilage degradation in an individual.
18. 18. The method of claim 16, wherein the administration step is parenteral or oral.
19. 19. A method for treating a bone or cartilage disorder in an individual in need thereof, this method comprises administering to the individual an effective amount of the complex of claim 7.
20. 20. The method of claim 19, wherein the bone disorder is chosen from the group of osteoporosis, osteogenesis imperfecta and bone fractures.
21. 21. The method of claim 18, wherein the cartilage disorder, is osteoarthritis, inflammatory arthritis, a torn tendon or a torn ligament.
22. 22. The method of claim 19, wherein the administration is parenteral or oral.
23. 23. The use of the complex of claim 7, to treat a bone or cartilage disorder, in an individual.
24. 24. A pharmaceutical formulation that opposes bone demineralization and that inhibits cartilage degradation, this formulation comprises: (a) a therapeutically effective amount of the complex of claim 7; and (b) a pharmaceutically acceptable carrier or diluent thereof.
25. 25. A method for treating a mammal to alleviate the pathological effects of osteoporosis, osteogenesis imperfecta, bone fractures, osteoarthritis and other bone and cartilage disorders, wherein this method comprises administering to the mammal the complex of claim 7, wherein the The complex is administered to this mammal in an amount sufficient to oppose the demineralization of the bone and the degradation of the cartilage.