CN1731985A - Sustained release l-arginine formulations and methods of manufacture and use - Google Patents

Abstract

The present invention provides methods and formulations for the treatment and prevention of cerebrovascular and cardiovascular diseases and disorders. The present invention is based, at least in part, on the discovery that administering to a subject a formulation comprising an agonist of endothelial nitric oxide synthase (eNOS), such as an HMG-CoA reductase inhibitor, and a formulation comprising a precursor of NO, such to as L-arginine, may be used to treat or prevent cerebrovascular and/or cardiovascular diseases or disorders.

Description

Sustained release l-arginine formulations and production thereof and using method

Related application

The title that the application requires on October 24th, 2002 to submit to is the U.S. Provisional Patent Application sequence number 60/421 of " method and composition of treatment cerebrovascular and cardiovascular disease and imbalance ", 258, the title of JIUYUE in 2003 submission on the 29th is the U.S. Provisional Patent Application sequence number 60/507 of " method and composition of treatment cerebrovascular and cardiovascular disease and imbalance ", 312, the title of submitting to on October 17th, 2003 is the U.S. Provisional Patent Application sequence number 60/XXX of " sustained release l-arginine formulations and production thereof and using method ", the priority of XXX, the complete content of each application above-mentioned are all incorporated into this paper as a reference by complete.

Background of invention

An enzyme family that is called as nitricoxide synthase (NOS) is from the synthetic nitric oxide (NO) of L-arginine-a kind of important second message,second messenger's biology.NOS has several different isotypes, comprises composition NOS (cNOS) and derivable NOS (iNOS).Two kinds of different cNOS are arranged: epidermis NOS (eNOS) and neuronal NOS (nNOS).ENOS participates in smooth muscle loosening, blood pressure reduces and the adjusting of the inhibition of platelet aggregation.ENOS is present in endotheliocyte and at responsive cell Ca ²⁺Receptor-mediated increase in the NO of release in a short time.People such as Michel, " nitric oxide is synthetic: which, where, how and why? " J.Clin.Invest 100:2146-2152 (1997).NNOS is important for long-term enhancing, and the dependence Ca of responsible nerve unit ²⁺Release.INOS works in host defense, and produce by activatory macrophage during the immunne response, in vascular smooth muscle cell, induce (for example, by various cytokines, microbial product, and/or bacterial endotoxin), and in a single day express just long-time synthetic NO.

Think in the endotheliocyte that forming nitric oxide by cNOS regulates, prevents that endothelial function is unusual plays an important role in as hyperlipemia, arteriosclerosis, thrombosis and restenosis in normal arterial pressure.CNOS is the main synthase that exists in brain and the epithelium, on function, cNOS has activity and can further stimulate by the increase of intracellular Ca2+ under basic condition, wherein the increase of the intracellular Ca2+ agonist or the Calcium ionophore of replying receptor-mediation takes place.CNOS seemingly this enzyme " physiology " form and in various groups of biological process, work.In vitro study shows that nitric oxide self can regulate the activity of NOS in degenerative mode.In the circulation of heart and brain kidney blood vessel, think that the main target of the NO that composition produces is soluble guanylate cyclase, it is arranged in vascular smooth muscle, cardiac muscle (myocyte) and coronary vasodilator smooth muscle.

Compare with cNOS, isotype iNOS derivable, that rely on calcium only describes in macrophage at first.The inducing of it is now know that nitricoxide synthase can be replied the adequate stimulus in many other cell types and be taken place.This is induced and can as taking place in the vascular smooth muscle cell, can also take place in the cell such as the cardiac muscle of expressing high-caliber composition isotype at the cell of not expressing the composition form of nitricoxide synthase usually.

INOS demonstrates negligible activity under basic condition, but when the factor of replying such as lipopolysaccharide and some cytokine, expresses in several hours time.The inductive form of this enzyme produces the NO amount more much bigger than composition form, and derivative NOS " Pathophysiology " form of this enzyme seemingly, because the NO pair cell that high concentration iNOS produces is deleterious.Can suppress inducing of iNOS by glucocorticoid and some cytokine.Know less relatively regulated in the back of transcribing about iNOS.The cytotoxic effect that NO causes may be independent of guanylate cyclase to a great extent and cyclo GMP forms.Major part research in this field concentrates on uses the arginic various derivants of L-to stimulate the iNOS inhibitor.

NO is from molecular oxygen and the synthetic metastable free radical of the arginic guanidine radicals nitrogen of L-in the catalytic reaction of NOS.Find this enzyme in many tissues and cell type, these cell types comprise the endotheliocyte of neuron, macrophage, hepatocyte, smooth muscle cell, blood vessel and the epithelial cell of kidney.NO acts near its point of release, enters target cell and activates cytosol enzyme guanylate cyclase, the formation of guanylate cyclase catalysis second message,second messenger cyclo GMP (cGMP).In the several seconds that NO forms, its experience is oxidized to nitrite or nitrate.David L.Nelson, Michael M.Cox, Lehninger Principles of Biochemistry, 892 pages, third edition Worth Publishers, 2000.

Replying various vasoactive agents, even during physiological stimulation, endotheliocyte discharges a kind of short-lived vasodilation that is called the deutero-relaxation factor of endotheliocyte (EDRF) and (is also referred to as the nitric oxide (EDNO) in endotheliocyte source.The product of inflammation and platelet aggregation such as 5-hydroxy tryptamine, histamine, Kallidin I, purine and thrombin are by stimulating their all or part effect of release performance of NO.The mechanism of lax dependence endotheliocyte comprises in the coronary circulation it being important in various vascular bedes.People such as Hobbs, Annu.Rev.Pharmacol.Toxicol.39:191-220 (1999).NO can easily be diffused into following smooth muscle and increase cGMP concentration by the activation guanylate cyclase, thereby the induction of vascular smooth muscle is lax.

NO is responsible for relying on neurotransmission and activatory macrophage cytotoxicity in activation, maincenter and the peripheral nervous system of lax and soluble guanylate cyclase of endotheliocyte.In vasculature, EDNO has several effects, comprising the inhibition of platelet aggregation, the adhesion of inflammatory cell and the propagation of smooth muscle cell.Particularly, EDNO is the important regulator of vascular tone.And, rely on the mediation that mobile diastole (a kind of endothelial function index commonly used) is subjected to NO to a great extent.

To the stimulation of the endotheliocyte of liner vasculature, as acetylcholine, Kallidin I, shearing force, or the like start the regulation mechanism of NO to vascular tone.Catalytic activity by contained eNOS in these endotheliocytes produces NO from the L-arginine.The NO that is produced leaves endotheliocyte and stimulates guanylate cyclase activity in the contiguous smooth muscle cell.The activation of guanylate cyclase has increased the cGMP level and has caused smooth cell lax, thus blood vessel dilating and increasing blood flow.People such as Moncada, New Eng.J.Med.329:2002-2012 (1993); People such as Vallance, J.Royl.Coll.Physician London 28:209-219 (1994).

Summary of the invention

The invention provides the method for treatment and prevention angiopathy and imbalance, these angiopathys and imbalance include, but not limited to cardiovascular, cerebrovascular and peripheral blood vessel and imbalance.The present invention to small part based on following discovery, promptly the arginic slow releasing preparation of HMG-CoA reductase inhibitor and L-is applied in treatment and prevention angiopathy and imbalance jointly, especially has synergy in cholesterol reducing and the triglyceride.In addition, the invention provides arginic slow releasing preparation of L-and production method, said preparation and production method make compositions have best release profile figure.In addition, said preparation and production method make compositions to be compressed easily, but can too brittle.

On the one hand, the invention provides the method that reduces cholesterol among the experimenter, this method comprises experimenter's administration of HMG-CoA reductase inhibitor and contains the arginic slow releasing preparation of L-.In various embodiments, this method hypercholesterolemia reducing, low density lipoprotein, LDL (LDL) cholesterol and/or triglyceride levels.In addition, this method has increased high density lipoprotein (HDL) cholesterol.And, this method hypercholesterolemia reducing, LDL cholesterol and/or triglyceride levels, and/or the degree that increases the HDL level not use the arginic degree of L-than administration of HMG-CoA reductase inhibitor only bigger.

On the other hand, the invention provides by administration of HMG-CoA reductase inhibitor and L-arginine and make the method that NO production increases among the experimenter that asymmetric diethylarginine (ADMA) raises.On the other hand, the invention provides the enhanced method of experimenter's medium vessels diastole that asymmetric diethylarginine (ADMA) is raise by administration of HMG-CoA reductase inhibitor and L-arginine.In the various embodiments in these areas, the L-arginine exists as slow releasing preparation.In other embodiments, the HMG-CoA reductase inhibitor is a simvastatin.In certain embodiments, the experimenter to suffer from endothelial function unusual.In other embodiments aspect these of the present invention, this method strengthens endothelial function.

On the other hand, the invention provides by the experimenter being used the method that the L-arginine makes the increase that nitric oxide (NO) produces among the experimenter that this asymmetric diethylarginine (ADMA) raises, wherein the L-arginine has overcome the inhibition effect of ADMA.On the other hand, the invention provides by the experimenter being used the enhanced method of experimenter's medium vessels diastole that the L-arginine raises this asymmetric diethylarginine (ADMA), wherein the L-arginine has overcome the inhibition effect of ADMA.In the various embodiments aspect these of the present invention, HMG-CoA reductase inhibitor (for example, simvastatin) is used jointly with the L-arginine.In certain embodiments, the L-arginine exists as slow releasing preparation.In the present invention's other embodiments aspect these, this method has strengthened endothelial function.

On the other hand, the invention provides the sustained release l-arginine compositions, it comprises about by weight 25% to about 75%L-arginine or its pharmaceutically acceptable salt; About by weight 0.5% to about 5% polyvinylpyrrolidone; About by weight 5% to about 40% hydroxypropyl emthylcellulose; About by weight 2% to about 20% microcrystalline Cellulose; About by weight silicon dioxide and about by weight magnesium stearate below 3% below 3%.In specific embodiments, said composition comprises about by weight 50%L-arginine mono-hydrochloric salts, and wherein the L-arginine is a L-arginine mono-hydrochloric salts; About by weight 3% to about 4% polyvinylpyrrolidone; About by weight 35% hydroxypropyl emthylcellulose; About by weight 10% microcrystalline Cellulose; About by weight colloidal silica below 1%, wherein silicon dioxide is colloidal silica; About by weight magnesium stearate below 1%.

On the other hand, the invention provides the method for preparing the arginic slow releasing preparation of L-, this method comprises with granulating agent granulation L-arginine to form granule; Moistening this granule that grinds; Dry granule; Drying grinds granule; And this granule mixed with at least a slow releasing preparation.In various embodiments, blend step can comprise premixing, mixing and final mixing granules agent.In another embodiment, this method can comprise that granulation step is preceding with L-arginine and bonding agent dry mixed.This bonding agent can be a polyvinylpyrrolidone.

In the specific embodiments of the present invention aspect this, this method can comprise granulation L-arginine, wherein the L-arginine is the about 50% of slow releasing preparation by weight, and granulating agent comprises polyvinylpyrrolidone, and wherein polyvinylpyrrolidone is about 3% to about 4% of slow releasing preparation by weight; Moistening this granule that grinds; Dry granule; Drying grinds granule; And this granule mixed with hydroxypropyl emthylcellulose, wherein hydroxypropyl emthylcellulose is about 35% of slow releasing preparation by weight.This method can also comprise mixes granule with microcrystalline Cellulose, colloidal silica and magnesium stearate, wherein microcrystalline Cellulose is about 10% of slow releasing preparation by weight, wherein colloidal silica is the about below 1% of slow releasing preparation, and wherein magnesium stearate is the about below 1% of slow releasing preparation by weight.

On the other hand, the invention provides and contain the arginic slow releasing preparation of L-() food stick for example, the arginic slow-releasing granules of L-, it is used for the treatment of or prevents angiopathy or imbalance.This food stick can also contain HMG-CoA reductase inhibitor (for example, simvastatin).In various embodiments, this food stick cholesterol reducing, reduction C-reactive protein can treat or prevent Alzheimer, and/or can treat or prevent intermittent claudication.

On the other hand, the invention provides the method for prevention or treatment experimenter's medium vessels disease or imbalance, this method comprises uses the food stick that contains L-arginine slow releasing preparation to the experimenter.More on the one hand, the invention provides the method that reduces cholesterol among the experimenter, this method comprises uses the food stick that contains L-arginine slow releasing preparation to the experimenter.More on the one hand, the invention provides nitric oxide production method among the increase experimenter, this method comprises uses the food stick that contains L-arginine slow releasing preparation to the experimenter.On the other hand, the invention provides the method that increases the diastole of experimenter's medium vessels, this method comprises uses the food stick that contains L-arginine slow releasing preparation to the experimenter.On the other hand, the invention provides the method for Alzheimer among treatment or the prevention experimenter, this method comprises uses the food stick that contains L-arginine slow releasing preparation to the experimenter.In other respects, the invention provides the method for treatment or the limping of prevention experimenter discontinuous, this method comprises uses the food stick that contains L-arginine slow releasing preparation to the experimenter.More on the one hand, the invention provides the method that reduces proteins C reactive among the experimenter, this method comprises that the experimenter uses the food stick that contains L-arginine slow releasing preparation.In some embodiments aspect front of the present invention, food stick can also contain HMG-CoA reductase inhibitor (for example, simvastatin).

On the other hand, the invention provides the method that reduces cholesterol among the experimenter, this method comprises uses L-arginine slow releasing preparation to the experimenter.In various embodiments, this method can reduce T-CHOL, low density lipoprotein, LDL (LDL) cholesterol among the experimenter, and/or triglyceride, and/or increases high density lipoprotein (HDL) cholesterol.In other respects, the invention provides the method for treatment or prevention Alzheimer, this method comprises uses L-arginine slow releasing preparation to the experimenter.More on the one hand, the invention provides the method for treatment or the limping of prevention discontinuous, this method comprises uses L-arginine slow releasing preparation to the experimenter.More on the one hand, the invention provides the method that reduces proteins C reactive, this method comprises that the experimenter uses L-arginine (for example, sustained release l-arginine).In some embodiments aspect front of the present invention, slow releasing preparation comprises about by weight 25% to about 75% L-arginine or its pharmaceutically acceptable salt; About by weight 0.5% to about 5% polyvinylpyrrolidone; About by weight 5% to about 40% hydroxypropyl emthylcellulose; About by weight 2% to about 20% microcrystalline Cellulose; About by weight silicon dioxide and about by weight magnesium stearate below 3% below 3%.In specific embodiments, said composition comprises about by weight 50%L-arginine mono-hydrochloric salts, and wherein the L-arginine is a L-arginine mono-hydrochloric salts; About by weight 3% to about 4% polyvinylpyrrolidone; About by weight 35% hydroxypropyl emthylcellulose; About by weight 10% microcrystalline Cellulose; About by weight colloidal silica below 1%, wherein silicon dioxide is colloidal silica; About by weight magnesium stearate below 1%.

Various aspect other, the invention provides the atherosis method of method, treatment or the prevention of arterial of treatment or prevention angiopathy or imbalance, increase vasodilative method, and/or the method for increase nitric oxide generation, this method comprises uses slow releasing preparation to the experimenter, and this slow releasing preparation comprises about by weight 25% to about 75%L-arginine or its pharmaceutically acceptable salt; About by weight 0.5% to about 5% polyvinylpyrrolidone; About by weight 5% to about 40% hydroxypropyl emthylcellulose; About by weight 2% to about 20% microcrystalline Cellulose; About by weight silicon dioxide and about by weight magnesium stearate below 3% below 3%.In the specific embodiments of aspect, said composition comprises about by weight 50%L-arginine mono-hydrochloric salts in front, and wherein the L-arginine is a L-arginine mono-hydrochloric salts; About by weight 3% to about 4% polyvinylpyrrolidone; About by weight 35% hydroxypropyl emthylcellulose; About by weight 10% microcrystalline Cellulose; About by weight colloidal silica below 1%, wherein silicon dioxide is colloidal silica; About by weight magnesium stearate below 1%.

On the other hand, the invention provides the method that reduces proteins C reactive among the experimenter, this method comprises experimenter's administration of HMG-CoA reductase inhibitor and the arginic slow releasing preparation of L-.This method and administration of HMG-CoA reductase inhibitor only, or only use the L-arginine and compare and reduce proteins C reactive among the experimenter to a greater degree.

According to following detailed and claim, other features and advantages of the present invention will be tangible.

The accompanying drawing summary

Fig. 1 is the curve chart of release mode of describing to contain the preparation of L-arginine and simvastatin.

Fig. 2 is the NMR image photograph with infarction size in the mouse brain of L-arginine and simvastatin processing and the untreated mouse brain.

Fig. 3 is the bar diagram of Infarction volume in the mice of describing to handle with L-arginine, simvastatin and L-arginine and simvastatin.

Fig. 4 is the bar diagram of Infarction volume in the mice of describing to handle with the simvastatin of L-arginine and various levels.

Fig. 5 is a flow chart of describing to produce the method for sustained release l-arginine tablet.

Fig. 6 is a flow chart of describing to produce the method for sustained release l-arginine tablet.

Fig. 7 is the bar diagram of the performance of comparison sustained release l-arginine formulations.

Fig. 8 be the comparison simvastatin with or not with the figure that philtrum is relied on the vasodilative influence of endothelium that uses of sustained release l-arginine compositions of the present invention.

Fig. 9 is the synergistic figure to the philtrum cholesterol levels of using of general introduction simvastatin and sustained release l-arginine compositions of the present invention.

Figure 10 illustrates the bar diagram of simvastatin to the influence of the HAEC of the human aorta endotheliocyte (HAEC) cultivated and untreated cultivation.

Detailed Description Of The Invention

The invention provides the method for the treatment of and prevention vascular diseases and imbalance, that these vascular diseases and imbalance include, but not limited to is cardiovascular, the cerebrovascular and peripheral artery disease and imbalance. The present invention is at least part of based on following discovery, the sustained release preparation that is HMG-CoA reductase inhibitor and L-arginine is applied in treatment and prevention vascular diseases and imbalance (comprising the cerebrovascular, cardiovascular and peripheral artery disease and imbalance) jointly, especially reduces the cholesterol aspect and has synergy. In addition, sustained release l-arginine and randomly, the HMG-CoA reductase inhibitor can be used for increasing vasodilation, increases NO and produce and reduce the C-proteins C reactive. In another embodiment, preparation described herein and method can be used for delaying the outbreak of this disease, imbalance and/or event in for example being in the colony that takes place in the danger that vascular diseases or imbalance and/or event take place. HMG-CoA reductase inhibitor and L-arginine sustained release preparation can be sequentially or are applied to simultaneously this experimenter. This reductase inhibitor and L-arginine can be included in the unitary agent.

In addition, the invention provides sustained release preparation and the production method of L-arginine, this sustained release preparation and production method are so that composition has best release profile figure. In addition, said preparation and production method so that composition can be compressed easily, but can too brittle.

In one embodiment, the preparation that is used for method of the present invention contains at least a sustained release agent (for purposes of the invention, controlled release and slowly-releasing can Alternates), for example, at least a slowly-releasing activator of endothelial nitric oxide synthase (for example, HMG-CoA reductase inhibitor and/or nitric oxide precursors are such as L-arginine). In another embodiment, L-arginine is slowly released in experimenter's the system. The pharmacokinetics collection of illustrative plates of the L-arginine that the slowly-releasing of L-arginine produces in blood plasma, this pharmacokinetics collection of illustrative plates provide the supply of the substantial constant that produces the required L-arginine of NO for NOS. Therefore, said preparation can dissolve and discharge the basically consistent amount of L-arginine lentamente in vivo in a period of time, its for the experimenter be treatment effectively. In another embodiment, the HMG-CoA reductase inhibitor is slowly released in experimenter's the system. In an embodiment again, being created in long-time of NO is basically consistent.

In another aspect of this invention, provide with the form of food and to be used for the treatment of vascular diseases and (to comprise, but be not limited to cardiovascular, the cerebrovascular, peripheral artery disease and imbalance), the composition of Charcot's syndrome, crucial limb ischaemic and Alzheimer's. These compositions of food form also can be used for increasing vasodilation, increase the NO generation and reduce cholesterol. Preferably, food is rod type food, as the form of the healthy rod type food of writing out a prescription. Use food so that the amount of the L-arginine that can provide is bigger than the amount that can mix single tablet. The invention provides rod type food, it can provide the 1 above L-arginine of gram and other reagent (if hope). In one embodiment, L-arginine is as quick releasing formulation, and for example the quick-release granula of L-arginine joins in the food bar. In another embodiment, rod type food comprises sustained release preparation, and it comprises, for example, and the slowly-releasing granula of L-arginine. In another embodiment, rod type food also contains extra reagent, such as the HMG-CoA reductase inhibitor. Preferably, the HMG-CoA reductase inhibitor is statins, such as Simvastatin.

Definition

Before further describing the present invention, for convenience's sake, collected some term that is used for this specification, embodiment and claims here.

As used herein, unless otherwise noted, term " experimenter " comprises mammal. Term " mammal " includes, but not limited to dog, cat, ox, horse, pig, and the people.

As used herein, term " treatment " etc. refers to the patient is used or the administering therapeutic agent, perhaps to organizations or administering therapeutic agent or preparation from patient's separation, this patient suffers from disease or imbalance, the symptom of disease or imbalance, the perhaps tendency of disease or imbalance, purpose is to cure, treat, alleviate, remove, change, remedy, prevent, improve, postpone the outbreak of disease or imbalance and/or event, delay the deterioration of disease or imbalance, improve or affect the symptom of this disease or imbalance, disease or imbalance, perhaps the inducement of disease or imbalance and/or event.

As used herein, term " vascular diseases " or " blood vessel imbalance " are often referred to the disease of blood vessel or imbalance and include, but not limited to cardiovascular, the cerebrovascular and peripheral artery disease or imbalance. Angiocardiopathy refers to the disease of the blood vessel of heart. See, for example, Kaplan, R.M. waits the people, " angiocardiopathy ", Health and Human Behavior, 206-242 page or leaf (McGraw-Hill, New York 1993). Angiocardiopathy is one of several forms simultaneously, comprises, for example, hypertension (being also referred to as high blood presses), coronary heart disease, apoplexy, and rheumatic heart disease. Peripheral artery disease or imbalance refer to the disease of the outer any blood vessel of heart. For example, peripheral artery disease can refer to that the blood vessel that blood is carried to leg and arm muscle narrows down. Cranial vascular disease refers to affect blood vessel to the disease of the ability of brain blood supply.

Term " atherosclerotic " is included in doctor's identification of obtaining employment in the medical science association area and the vascular diseases of understanding, imbalance and illness. Atherosclerotic cardiovascular disease, coronary heart disease (being also referred to as coronary artery disease or ischemic heart disease), cerebrovascular disease and peripheral vascular disease all are atherosclerotic clinical manifestations and therefore included by term " atherosclerotic " and " Atheromatosis ".

Term " is jointly used " when be used for describing and is referred to that the compound that can use by identical or different approach is by simultaneously (for example when the experimenter used two or more compounds as used herein, as mixture) or use in proper order, thereby the pharmacological effect of every kind of compound is in time overlapping. As used herein, unless otherwise indicated, when being applied to the using of at least two kinds of compounds, term " sequentially " refers to administered compound so that the pharmacological effect of every kind of compound is in time overlapping. In certain embodiments, reagent is used basically simultaneously. It is upper enough approaching that " basically side by side " refers to that preparation of the present invention is applied to time that is applied in of experimenter and at least a extra reagent, thereby these reagent can bring into play addition or even collaborative effect, such as, but not limited to, increase NOS activity, NO generation, or vasodilation.

As used herein, term " precursor of NO " comprises any substrate precursor of natural NO, for example, and 1B.

Term " natural NO " refers to the bio-transformation by L-arginine or relies on the nitric oxide that the approach of L-arginine produces as used herein. Term " relaxation factor in endothelium source (EDRF " or " endothelial cell source nitric oxide (EDDO) " can with " natural NO " Alternate.

Term " L-arginine " refers to L-arginine and all its biochemical equivalents as used herein, for example, L-arginine monohydrochloride, precursor and its alkaline form, they are as the substrate of NOS, and causing NO to produce increases.This term comprises the arginic pharmaceutically acceptable salt of L-.

Term " pharmaceutically acceptable salt " refers to that these non-toxic acids or alkali comprise inorganic bronsted lowry acids and bases bronsted lowry or organic bronsted lowry acids and bases bronsted lowry from the salt of pharmaceutically acceptable non-toxic acid or alkali preparation.Suitable non-toxic acid comprises inorganic and organic acid, as acetic acid, benzenesulfonic acid, benzoic acid, camphorsulfonic acid, citric acid, ethyl sulfonic acid, fumaric acid, gluconic acid, glutamic acid, hydrobromic acid, hydrochloric acid, isethionic acid, lactic acid, maleic acid, malic acid, mandelic acid, methanesulfonic acid, glactaric acid, nitric acid, pamoic, pantothenic acid, phosphoric acid, succinic acid, sulphuric acid, tartaric acid, right-toluenesulfonic acid, or the like.Especially preferred hydrochloric acid, hydrobromic acid, phosphoric acid and sulphuric acid, the most especially preferred salt hydrochlorate.

Because be used for the L-arginine of the inventive method is alkalescence and tart, so can prepare salt from pharmaceutically acceptable non-toxic acid or alkali, these acid or alkali comprise inorganic and organic acid or inorganic and organic base.These salt can contain following anionic any: acetate, benzenesulfonic acid root, benzoate anion, camphorsulfonic acid root, citrate, fumarate, glucose acid group, chloride ion, lactate, maleate, glactaric acid root, nitrate anion, embonate, phosphate radical, amber acid radical, sulfate radical, tartrate anion, or the like.Especially preferred benzenesulfonic acid root, bromide ion, chloride ion and sulfate radical.These salt can also contain following cation: aluminum, calcium, lithium, magnesium, potassium, sodium, zinc, benzyl star (benzathine), chloroprocaine, choline, diethanolamine, ethylenediamine, meglumine and procaine.

As used herein term " agonist " or " agonist of eNOS or cNOS " refer to stimulate substrate as, for example, the L-arginine is to the reagent of the biotransformation of NO.The agonist of eNOS or cNOS comprises, for example, and the HMG-CoA reductase inhibitor." HMG-CoA reductase (3-hydroxy-2-methyl glutaryl-coenzyme A) " is the microbial enzyme of the rate-limiting reaction in the catalysis cholesterol biosynthesis." HMG-CoA reductase inhibitor " suppresses the HMG-CoA reductase.The HMG-CoA reductase inhibitor also is known as " statins ".

Described the natural or synthetic chemical compound lot that obtains in the document, they suppress the HMG-CoA reductase and are called as " statins ", and are formed for implementing reagent classification of the present invention.Example include, but not limited to by commercial sources available those, as simvastatin (U.S. Patent number 4,444,784), lovastatin (U.S. Patent number 4,231,938), pravastatin sodium (U.S. Patent number 4,346,227), Fluvastatin (U.S. Patent number 4,739,073), his spit of fland (U.S. Patent number 5,273 is cut down in holder, 995), simvastatin, rosuvastatin, with many other, such as mevastatin, dalvastatin, mevastatin, fluvastatin, pitavastatin, HR-780, GR-95030, CI 980, BMY 22089, BMY 22566 and for example, U.S. Patent number 5,622,985, U.S. Patent number 5,135,935, U.S. Patent number 5,356,896, U.S. Patent number 4,920,109, U.S. Patent number 5,286,895, U.S. Patent number 5,262,435, U.S. Patent number 5,260,332, U.S. Patent number 5,317,031, U.S. Patent number 5,283,256, U.S. Patent number 5,256,689, U.S. Patent number 5,182,298, U.S. Patent number 5,369,125, U.S. Patent number 5,302,604, U.S. Patent number 5,166,171, U.S. Patent number 5,202,327, U.S. Patent number 5,276,021, U.S. Patent number 5,196,440, U.S. Patent number 5,091,386, U.S. Patent number 5,091,378, U.S. Patent number 4,904,646, U.S. Patent number 5,385,932, U.S. Patent number 5,250,435, U.S. Patent number 5,132,312, U.S. Patent number 5,130,306, U.S. Patent number 5,116,870, U.S. Patent number 5,112,857, U.S. Patent number 5,102,911, U.S. Patent number 5,098,931, U.S. Patent number 5,081,136, U.S. Patent number 5,025,000, U.S. Patent number 5,021,453, U.S. Patent number 5,017,716, U.S. Patent number 5,001,144, U.S. Patent number 5,001,128, U.S. Patent number 4,997,837, U.S. Patent number 4,996,234, U.S. Patent number 4,994,494, U.S. Patent number 4,992,429, U.S. Patent number 4,970,231, U.S. Patent number 4,968,693, U.S. Patent number 4,963,538, U.S. Patent number 4,957,940, U.S. Patent number 4,950,675, U.S. Patent number 4,946,864, U.S. Patent number 4,946,860, U.S. Patent number 4,940,800, U.S. Patent number 4,940,727, U.S. Patent number 4,939,143, U.S. Patent number 4,929,620, U.S. Patent number 4,923,861, U.S. Patent number 4,906,657, U.S. Patent number 4,906,624 and U.S. Patent number 4,897, those reagent of describing in 402, these patents all are incorporated by reference herein.Can also use the combination of two or more HMG-CoA reductase inhibitors in the method for the present invention.

Term " eNOS activity " phalangeal cell produces the ability of NO from substrate L-arginine as used herein.Can realize the active increase of eNOS with many diverse ways.For example, the increase of the increase of the proteic amount of eNOS or this protein active (yet keep this proteinic constant level) can cause " activity " that increase.The increase of available proteinic amount from, for example and be not limited to, the translation of the increase of the transcribing of the increase of eNOS gene, eNOS mRNA, the stability of the increase of eNOSmRNA, the activation of eNOS, or the minimizing of eNOS protein degradation.

Also can measure eNOS activity in the cell or tissue with many distinct methods.A kind of direct measurement is an amount of measuring the eNOS that exists.Another kind of directly the measurement is to measure by eNOS to make the L-arginine to the amount of citrulline conversion or at specified conditions, as the nitric oxide production amount that produces by eNOS under the physiological condition of tissue.Can also measure the eNOS activity indirectly, for example by measuring mRNA half life (upstream indicator) or replying (downstream indicator) by the phenotype that NO is existed and to implement this indirect measurement.A kind of phenotype measurement that is used for this area is measure the dependence endothelium reply acetylcholine lax, and this is replied and is subjected to the active influence of eNOS.The NO level of utilizing NO meter to exist in can measuring samples.The technology of all fronts all is that those skilled in the art know.

Method of the present invention not only allows to rebuild the active normal baseline level of eNOS by the increase that causes the NO generation, and allows to increase this activity that is higher than the normal baseline level.The normal baseline level is the active amount in the normal control group, and it is subjected to the control at age and does not indicate endothelial cell nitric oxide synthase activity to change the symptom of (as hypoxia condition, hyperlipemia, or the like).Practical level will depend on selected concrete age group and be used to assess active concrete measuring method.In unusual environment, endothelial cell nitric oxide synthase activity (producing with NO) is reduced to below the normal level.Therefore, preparation of the present invention not only can recover the normal baseline level that NO in these abnormal conditiones produces, and endothelial cell nitric oxide synthase activity (producing with NO) is increased to be higher than far away more than the normal baseline level.

Term " carrier " refers to be used for diluent, the excipient of the mixture of pharmaceutical compositions, or the like.

As used herein, term " dosage form " refers to pharmaceutical composition, and it contains to be useful on single or multiple dosage and is applied to the experimenter, for example the Sq of patient's active component.

Term " mg/Kg " refers to the mg of every Kg subject retry agent as used herein.

As used herein, unless otherwise indicated, term " half life " refers to that the concentration of biological blood plasma Chinese medicine is reduced to half time that is spent of pact of this drug level when using.

As used herein, unless otherwise indicated, term " discharges immediately " and refers to not have extrinsic factor to postpone the release in vitro of one or more medicines.

As used herein, use is exchanged in term " pharmaceutical composition " or " pharmaceutical preparation " herein, refers to contain the compositions of pharmaceutically acceptable composition.

Term " pharmaceutically acceptable " refers to a kind of preparation type as used herein, the said preparation type will by the administrative organization of federation or state government evaluation and may by or in American Pharmacopeia or other pharmacopeia of generally acknowledging, list and be used for animal, more specifically be used for the people.

As used herein, unless otherwise indicated, term " pharmaceutically acceptable carrier " refers to a kind of mounting medium, its not the interferon activity composition bioactive effectiveness and to its experimenter's avirulence of using.This medium of pharmaceutical active preparation or the purposes of reagent are to know in this area.Except with inconsistent conventional media of this reactive compound or reagent, consider to be used for to use arbitrary conventional media or reagent in the preparation of method of the present invention.

As used herein, term " pharmaceutically acceptable salt " refers to comprise mineral acid and organic acid from pharmaceutically acceptable non-toxic acid, the salt of preparation.

As used herein, unless otherwise indicated, term " slow release " is defined as the long-term release mode of one or more medicines, thereby medicine discharged in a period of time.For the present invention, slow release and controlled release are used interchangeably.

As used herein, unless otherwise indicated, term " salt or complex " is used to describe chemical compound or the compositions that contains two or more chemical parts, these two or more chemical parts are by the interaction combination of at least a type, these interactions comprise, but be not limited to Van der Waals force, ion and/or hydrogen bond.Salt or complex can be used as solid and exist or be present in the liquid.

The weight of the specific components of the weight of all components during term " percetage by weight " refers to based on said preparation when the amount of the component that is used for describing preparation as used herein.

Each side of the present invention is described in further detail in the trifle below:

I. be used for the treatment of or the method for prevention of brain blood vessel and cardiovascular disease and imbalance in preparation

Method of the present invention comprises treatment and prevention experimenter, the method of philtrum cerebrovascular and/or cardiovascular disease or imbalance for example, this method comprise simultaneously or in a sequence to be used the preparation that contains the HMG-CoA reductase inhibitor and contains the arginic preparation of L-the experimenter.Alternatively, the experimenter is used the unitary agent that contains L-arginine and HMG-CoA reductase inhibitor.

One embodiment of the invention comprise the L-arginine formulations of slow releasing preparation form, HMG-CoA reductase inhibitor preparation or the L-arginine of slow releasing preparation form and the preparation of HMG-CoA reductase inhibitor of slow releasing preparation form.In one embodiment, the present invention includes and contain the arginic preparation of L-, it can simultaneously or in a sequence be used with at least a HMG-CoA reductase inhibitor, and wherein said preparation concentration release L-arginine and HMG-CoA reductase inhibitor with substantial constant in long-time is present in the immediate release formulation.In another embodiment, the present invention includes and contain high concentration L-arginine and be the preparation of slow releasing preparation form, wherein the pharmacokinetics collection of illustrative plates is 0 grade of release dynamics (that is a linear in time rate of release).The release characteristic that can modify two class medicines is to provide release mode, and this release mode allows this to unite the single unit dose that is suitable for as once a day.

In one embodiment, the preparation that is used for the inventive method contains the L-arginine for the treatment of effective dose, the HMG-CoA reductase inhibitor and at least a slow releasing agent of treatment effective dose.

Said preparation can also comprise for use, preservation, the required extra composition of said preparation of modifying such as attractive in appearance.In one embodiment, preparation of the present invention also comprises bonding agent, filler and lubricant.In preferred embodiments, said preparation contains the sustained release l-arginine prescription, and this prescription comprises L-arginine, bonding agent, one or more slow releasing agents, fluidizer and slow releasing agent or lubricant.Said preparation can also contain filler and compression agent.Slow releasing preparation of the present invention is especially favourable, because their release collection of illustrative plates allows to use than discharging immediately or keeping identical levels of drugs by the lower dosage of the available slow releasing agent of commercial sources.Because using of sustained release l-arginine and statins can also increase statins, for example, the effectiveness of simvastatin has beneficial effect of equal value so the use of preparation of the present invention can also allow the more low dosage of statins.

Can obtain the L-arginine by the known many sources of technical staff.For example, by commercial sources from comprising that (Milwaukee, various approach WI) can obtain USP level L-arginine to Sigma-Aldrich.Suitable arginine and arginine derivative compound include, but not limited to arginine salt, as arginine monohydrochloride, arginine aspartate, or the arginine nicotinate.Other arginine compounds or derivant can be selected from dipeptides, it comprises arginine such as alanyl arginine (ALA-ARG), valyl-arginine (VAL-ARG), isoleucyl--arginine (ISO-ARG) and leucyl-arginine (LEU-ARG) and comprises arginic tripeptides such as arginyl-lysyl-glutamic acid (ARG-LYS-GLU) and arginyl-glycyl-arginine (ARG-GLY-ARG).The L-arginine is preferably L-arginine mono-hydrochloric salts.

In one embodiment, the L-arginine is about 10% to about 75% of preparation by weight.In another embodiment, the L-arginine is about 25% to about 75% of preparation by weight.In preferred embodiments, the L-arginine is about 50% of preparation by weight.

The use of one or more slow releasing agents allows slowly to discharge L-arginine and/or HMG-CoA reductase inhibitor in the longer time.For example, slow releasing agent will discharge the L-arginine with certain speed, and this speed will can not cause concentration peak or low ebb, and this concentration peak or low ebb will make the side effect aggravation relevant with the arginic high concentration of L-or low concentration in the blood flow.The slow releasing agent that is suitable for use in the preparation of method of the present invention comprises hydrating agents, and for example, such as cellulose, it is partially hydrated and form the gel barrier when contacting with aqueous environments, and this barrier hinders the dissolving of the reagent of quilt that hydrating agents wraps.In other words,, slow releasing agent slowly absorbed preparation thereby forming temporary transient water barrier water, thus hydration said preparation and subsequently with the much lower speed release of active ingredients of the preparation that does not have slow releasing agent, for example, the L-arginine.In addition, slow releasing agent is present in the granular size, and when this granule was impregnated in capsule or compacting or is compressed into tablet, pill or gelcap, water slowly was penetrated into this structure.

In one embodiment, one or more slow releasing agents include, but not limited to cellulose ether product, polymethyl methacrylate or polyvinyl alcohol.In another embodiment, slow releasing agent comprises cellulose, and it includes, but are not limited to methylcellulose, hydroxypropyl emthylcellulose, hydroxyethyl-cellulose or their combination.In preferred embodiments, slow releasing agent comprises one or more hydroxypropyl emthylcelluloses.Suitable slow releasing agent can obtain with trade (brand) name METHOCEL  and ETHOCEL  from The DowChemical Company by commercial sources.In preferred embodiments, slow releasing agent is METHOCEL  K100M CR Premium and/or METHOCEL  E4M CR Premium.

Slow releasing agent exists with release of active ingredients in the desirable time with enough amounts usually, for example, and L-arginine or HMG-CoA reductase inhibitor.In one embodiment, existing slow releasing agent is about 5% to about 40% of preparation by weight.In another embodiment, existing slow releasing agent is about 5% to about 75% by weight.In a further embodiment, existing slow releasing agent is about 15% to about 50% of preparation by weight.In preferred embodiments, existing slow releasing agent is about 35% of preparation by weight.All scopes in the top scope all within the scope of the invention.

In one embodiment, slow releasing agent discharged the L-arginine in 10 hours, as describing among Fig. 1.In one embodiment, slow releasing agent as one man discharged the L-arginine basically in about 4 hours to about 24 hours.In another embodiment, preparation of the present invention as one man discharged the L-arginine basically in about 8 hours to about 24 hours.In a further embodiment, sustained release l-arginine formulations as one man discharged the L-arginine basically in about 12 hours to about 48 hours.

In another embodiment, the preparation that is used for the inventive method will discharge arginine in one way so that the pharmacokinetics collection of illustrative plates to be provided, wherein half life (T _1/2) and T _MaxEnough keep the L-arginine on the level of substantial constant.In other words, in one embodiment,, slow releasing preparation of the present invention realized that the arginic stable state of circulation L-and this stable state keep constant thereby discharging the L-arginine.In one embodiment, the pharmacokinetics collection of illustrative plates is for making T _1/2Be about 4 hours to about 12 hours and T _MaxBe about 4 hours.In a further embodiment, T _1/2Be about 4 hours to about 8 hours and T _MaxBe about 4 hours.

Be used for bonding agent of the present invention and comprise generally well-known those bonding agent of technical staff.Bonding agent includes, but not limited to sugar, as lactose, sucrose, glucose, dextrose and molasses; Natural and synthetic natural gum are as extract, panwar natural gum and the ghatti natural gum of Radix Acaciae senegalis, guar gum, sodium alginate, Irish lichen; Other bonding agent comprise mixture, methylcellulose, sodium carboxymethyl cellulose, hydroxypropyl cellulose (HPC), hydroxyethyl-cellulose, hydroxypropyl emthylcellulose, alginic acid, ethyl cellulose, microcrystalline Cellulose, Carmomer, zein, starch, dextrin, maltodextrin, gelatin, pre-gelatinized starch, polyvinylpyrrolidone (PVP) or the polyvinyl pyrrolidone and their mixture of poly(ethylene oxide) and Polyethylene Glycol.In preferred embodiments, bonding agent is the polyvinylpyrrolidone homopolymer.

In one embodiment, existing bonding agent is the about below 20% of preparation by weight.In another embodiment, existing bonding agent is about 0.5% to about 5% of preparation by weight.In preferred embodiments, existing bonding agent is about 3% to about 4% of preparation by weight.

In preferred embodiments, sustained release l-arginine formulations also comprises fluidizer.Fluidizer can be any known UPS level fluidizer, comprises, for example, silicon dioxide.In preferred embodiments, fluidizer is a colloidal silica.

In one embodiment, existing fluidizer is the about below 3% of preparation by weight.In another embodiment, existing fluidizer is the about below 2% of preparation by weight.In preferred embodiments, existing fluidizer is the about below 1% of preparation by weight.

The filler that is used for preparation comprises generally well-known those filleies of technical staff.Typical filler includes, but not limited to sugar, as lactose, sucrose, dextrose, mannitol and sorbitol, milk surum, two alkali calcium phosphate, three alkali calcium phosphate, calcium sulfate and their mixture.Other filleies include but not limited to, cellulose preparation, such as corn starch, wheaten starch, rice starch, potato starch, gelatin, gum tragacanth, methylcellulose, hydroxypropyl emthylcellulose, sodium carboxymethyl cellulose, polyvinylpyrrolidone and their mixture.Microcrystalline Cellulose can also be as compression agent and filler.In preferred embodiments, filler/compression agent is a microcrystalline Cellulose.More preferably, microcrystalline Cellulose is sold with AVICEL  PH 102 titles by The Dow ChemicalCompany.

In one embodiment, existing filler is the about below 50% of preparation by weight.In another embodiment, existing filler is about 2% to about 20% of preparation by weight.In preferred embodiments, existing filler is about 10% of preparation by weight.

Can add excipient to increase the solid amount that exists in the preparation.Usually unite the excipient that is used for this purpose and comprise sodium phosphate or potassium, calcium carbonate, calcium phosphate, sodium chloride, citric acid, tartaric acid, gelatin, with carbohydrate such as dextrose, sucrose, lactose, sorbitol, inositol, mannitol and glucosan, starch, cellulose derivative, gelatin, and polymer, such as Polyethylene Glycol.Except pointing out to mention those, other be well known to a person skilled in the art.

The releasing agent or the lubricant that are used for said preparation comprise those that the technical staff is known usually.Typical lubricants comprises, but be not limited to, stearate, magnesium stearate, zinc stearate, calcium stearate, stearic acid, hydrogenated vegetable oil are (for example, the hydrogenation cottonseed oil), sodium stearyl fumarate, palmitostearate, Glyceryl Behenate, sodium benzoate, sodium lauryl sulphate, Stepanol MG, mineral oil, Talcum and their mixture.In preferred embodiments, lubricant is a magnesium stearate.In other embodiments, select lubricant to make and guarantee nutraceutical optimal absorption and utilization.

In one embodiment, existing lubricant is the about below 20% of said preparation by weight.In another embodiment, existing lubricant is about 2% to about 20% of said preparation by weight.In preferred embodiments, existing lubricant is about 10% of said preparation by weight.

Disintegrating agent comprises, but be not limited to sodium starch glycolate, cross-linked carboxymethyl cellulose sodium, crospovidone, crosslinked polyvinylpyrrolidone, corn starch, pre-gelatinized starch, microcrystalline Cellulose, alginic acid, phenol formaldehyde (PF) ion exchange resin, polyvinylpyrrolidone, polysaccharide, sodium carboxymethyl cellulose, agar, its salt such as sodium alginate, Primogel and their mixture.

The compression agent allows preparation to be shaped as tablet, lozenge, Jiao Gai (gelcap), perhaps with solid-state other forms of using.In one embodiment, the compression agent allows preparation to be shaped as tablet, lozenge or Jiao Gai.The compression agent includes, but not limited to Avicel, magnesium stearate, wax, natural gum, cellulose (celleusics), stearate, perhaps their associating.In preferred embodiments, the compression agent is a microcrystalline Cellulose.

In one embodiment, existing compression agent counts about 0.01% to about 5% by the percetage by weight of preparation.In another embodiment, the amount of existing compression agent is about 0.5% to about 3%.In a further embodiment, the amount of existing compression agent counts about 1% to about 2% by the percetage by weight of preparation.

In one embodiment, L-arginine prescription comprises the L-arginine in the unit dose, and it is enough for about 5mg/kg to about 40mg/kg experimenter's body weight.In another embodiment, L-arginine prescription comprises the L-arginine in the unit dose, and it is enough for about 20mg/kg to about 25mg/kg.

In another embodiment, L-arginine and HMG-CoA reductase inhibitor are all in slow releasing preparation.The amount of HMG-CoA reductase inhibitor can become based on the special inhibitor that exists in the preparation, because some inhibitor is more effective than other inhibitor.For example, the BAYCOL  that exists in each tablet can arrive about 0.8mg for about 0.1mg, and the ZOCOR  that exists in each tablet can arrive about 80mg for about 10mg.Those skilled in the art can determine therapeutic dose based on used special inhibitor.In one embodiment, the HMG-CoA reductase inhibitor is that simvastatin and existing unit dose are enough for about 0.5mg/kg to about 3mg/kg experimenter's body weight.In another embodiment, the HMG-CoA reductase inhibitor is that simvastatin and existing unit dose are enough for about 1.2mg/kg to about 1.4mg/kg experimenter's body weight.

In a further embodiment, L-arginine and HMG-CoA reductase inhibitor for example, provide in the independent tablet all at independent slow releasing preparation.Slow release HMG-CoA reductase inhibitor can be by commercial sources from for example, Merck ﹠amp; Company, (Rahway NJ) obtains Inc..

The preparation that is used for the inventive method can contain the pharmaceutical carrier of with good grounds conventional medicine complex technique.According to Orally administered desirable dosage form, this carrier can be taked various forms.In the preparation of preparation peroral dosage form, can use any conventional medicine medium.Most preferred oral solid preparation is tablet and gelcap.Alternatively, preparation of the present invention can mix in the capsule.In this embodiment, sustained release l-arginine granule and randomly, the HMG-CoA reductase inhibitor can be impregnated in the capsule.

Because they are easy to use, so tablet and capsule are represented best oral dosage unit form, use the solid-state drug carrier in this case.Tablet or capsule can contain the L-arginine formulations and the HMG-CoA reductase inhibitor preparation of different configurations in identical tablet or capsule.Configuration comprises, two parts half-sum half tablet or capsule, and around the preparation of another kind of preparation, the dispersant of a kind of preparation in another kind of preparation, two kinds of mutual blended granules of preparation, or the like.If wish, can peridium patch agent or capsule by standard aqueous or anhydrous technology.

The preparation that is used for the inventive method can also contain other pharmaceutically acceptable compositions, as composition commonly used in this area.See Remington:the Science ﹠amp; Practice ofPharmacy,, Alfonso R.Gennaro, the 20th edition, Williams ﹠amp; Wilkins, 2000.The extra composition that is used in the preparation of the inventive method comprises, but be not limited to, water, glycol, oils, ethanol, starch, sugar, diluent, disintegrating agent, antiseptic, excipient, lubricant, disintegrating agent, diluent, carrier, stabilizing agent, coloring agent, flavour enhancer and their associating.The example of suitable diluent comprises water, ethanol, polyhydric alcohol, vegetable oil, injectable organic ester such as ethyl oleate and their associating.Preparation can also contain adjuvant such as antiseptic, humidizer, emulsifying agent, and partitioning agent.By various antibacteriums and antifungal can guarantee the effect of prophylaxis of microbial, these antibacteriums and antifungal include but not limited to parabens, chlorobutanol, phenol, sorbic acid, or the like.Also wish to comprise isotonic agent, its include but not limited to sugar, sodium chloride, or the like.

In another embodiment of the invention, said preparation can also be used jointly with at least a other medicaments.The example of medicament classification comprises: adrenergic agent, Adrenocorticosteroids, the adrenal cortex inhibitor, aldosterone antagonist, aminoacid, ammonia detoxicant, anabolic agent, analeptic, analgesic, androgen, anesthetis, anoretics, antagonist, the antepituitary inhibitor, anthelmintic, anti-acne agent, antiadrenergic agent, anti--allergic agent, the agent of resistance to deformation worm, androgen antagonist, anti-anemia agent, the angor agent, antianxiety drug, the arthritis agent, anti-asthmatic agent, antiatherosclerotic, antibacterium, anti-cholelithiasis, anti-cholelithiasis forms, anticholinergic, anticoagulant, anticoccidiosis medicine, anticonvulsant, antidepressant, anti-glycosuria agent, diarrhea, antidiuretic, anti-emetic, epilepsy, estrogen antagonist, the fibrinolysis agent, antifungal, hemophilia, hemorrhage, antihistamine, hyperlipidemia disease, antihyperlipoproteinemic, hypotensive agent, infection, antiinflammatory, antikeratinizing agent, malaria, antimicrobial, migraine, resisting mitosis, antifungal, nausea, antitumor, antineutropenic, antiobesity agent, antiparasitic, anti-parkinson, antiperistaltic, anti-lung sac worm, antiproliferative, anti-prostate hyperplasia, protozoacide, itching relieving, psychosis, antirheumatic, schistosomicide, seborrhea, secretion inhibitor, spasmolytic, antithrombotic, cough medicine, antiulcer.Anti-urinary calculi agent; antiviral agent; appetite suppressant; the benign prostatauxe therapeutic agent; blood sugar regulator; bone resorption inhibitor; bronchodilator; carbonic anhydride alcohol inhibitor; cardiac sedative; cardioprotective agent; cardiac tonic; cardiovascular agents; choleretic; cholinergic drug; acetylcholine esterase passivator; the coccidiosis inhibitor; cognitive adjuvant; tranquilizer; diuretic; the dopaminergic agent; ectoparasitcide; emetic; enzyme inhibitor; estrogen; cellosolve; fluorescent agent; free oxygen free radical scavenger; the gastrointestinal motility effector; glucocorticoid; gonad-stimulation component; hair growth stimulant; hemorrhage.Histamine H2 receptor antagonist; hormone; hypocholia; hypoglycemia; hypolipemia; hypotension; developing agent; immunizing agent; immunomodulator; immunomodulator; immunostimulant; immunosuppressant; sexual impotence treatment adjuvant; keratolytic; the LNRII agonist; the liver imbalance is handled; luteolysin; the spirit performance enhancers; the mood regulation agent; mucolytic agent; the mucosa protective agent; mydriatic; nasal decongestant; neuromuscular blocking agents; the neuroprotective base; nmda antagonist; non-hormone sterols derivant; odinagogue; activator of plasminogen; platelet activating factor antagonist; platelet aggregation inhibitor; synergist; progesterone; prostaglandin; the prostate growth inhibitor; prothyrotropin; antipsychotic reagent; radiopharmaceutical agent; regulator; relafant; partitioning agent; antiscabietic; sclerosing agent; tranquilizer; selective adenosine A1 antagonist; 5-hydroxytryptamine antagonist; serotonin antagonist; the 5-hydroxytryptamine receptor antagonist; steroid; analeptic; inhibitor; symptomatic multiple sclerosis; synergist; thyroxin; thyroid imhibitor; thyromimetic; tranquilizer; the treatment of cerebral ischemia; the treatment of paget disease; the treatment of unstable angor; the uricosuric eccritic; vasoconstrictor; vasodilation; the wound agent; Wound-healing agent; perhaps xanthine oxidase inhibitor.

Another example of medicament comprises angiotensin converting enzyme inhibitor (ACE inhibitor).ACE is a kind of enzyme that catalysis angiotensin I transforms to angiotensin II.ACE inhibitor comprises aminoacid and its derivant, peptide, comprises the antibody of dipeptides and tripeptides and ACE, and these ACE inhibitor are disturbed the renin-angiotensin peptide by the activity that suppresses ACE, thereby reduces or eliminates the formation of precursor substance angiotensin II.ACE inhibitor medically is being used for the treatment of hypertension, congestive heart failure, myocardial infarction and nephropathy.The known compounds category that can be used as ACE inhibitor comprises acyl group sulfydryl and sulfydryl alkanoyl proline such as captopril (U.S. Patent number 4,105,776) and Zofenopril (U.S. Patent number 4; 316; 906); the carboxyalkyl dipeptides is as enalapril (U.S. Patent number 4,374; 829); lisinopril (U.S. Patent number 4; 374,829); quinapril (U.S. Patent number 4,344; 949); ramipril (U.S. Patent number 4; 587,258) and general indolic acid (U.S. Patent number 4,508; 729); the carboxyalkyl dipeptide analogue is as cilazapril (U.S. Patent number 4,512; 924) and benazepril (U.S. Patent number 4; 410,520); phosphinyl alkanoyl proline is as fosinopril (U.S. Patent number 4; 337,201) and trandolapril.Estrogen raises NOS and expresses, and ACE inhibitor does not influence expression, but influences the efficient of NOS to the effect of L-arginine.Thereby, can increase active with the whole bag of tricks.Usually, by inhibitor of the present invention by with respect to the cell of not handling according to reductase inhibitor according to the present invention, the increase of the amount of the organized enzyme that exists in the cell with this reductase processing and increase activity.

II. prevent and Therapeutic Method

On the one hand, the invention provides prevention experimenter's medium vessels disease or imbalance, method as cerebrovascular and/or cardiovascular disease or imbalance, this method comprise to the experimenter in the danger that is in cerebrovascular and/or cardiovascular disease or imbalance sequentially or simultaneously use contain the arginic preparation of L-with (for example contain the HMG-CoA reductase inhibitor, simvastatin) preparation perhaps contains the unitary agent of L-arginine and HMG-CoA reductase inhibitor.The experimenter who is in cerebrovascular and/or cardiovascular disease or the imbalance danger of (comprising incident) can pass through, for example, atherosclerotic inducement, atherosclerotic symptom, perhaps dangerous factor as, for example, smoking, hypertension, diabetes, family history, inherited genetic factors, elevated cholesterol, old age and drink and identify.

The using of preparation that is used for the inventive method as preventive used before can showing at the characteristic symptom of the outbreak of cerebrovascular and/or cardiovascular disease or imbalance, thereby, this cerebrovascular and/or cardiovascular disease or imbalance are prevented from, its deterioration is slowed down, and perhaps its outbreak postpones.

As at title for described in the International Patent Publication No. WO 00/56403 (it is incorporated into this paper as a reference by complete) of " raising III type endothelial cell nitric oxide synthase " by the HMG-CoA reductase inhibitor, the active rise of NOS is not depended on the synthetic minimizing of cholesterol and is not especially depended on the minimizing that ox-LDL forms.Therefore, it is active or increase in the infected cell or tissue thisly when active no matter when to wish to recover eNOS, can utilize the present invention.Tissue is defined as comprising to tissue provides cell in the vascular system of nutrition and the cell of expressing the tissue of eNOS.

Nitric oxide synthase activity is relevant with many diseases, and these diseases comprise sexual impotence, heart failure, harmonization of the stomach esophagus movement disorder, kidney imbalance, as kidney hypertension and carrying out property nephropathy, insufficient insulin, or the like.Suffering from the individuality of these diseases can be benefited from the increase that NO produces.For example, suffers from the individual nitric oxide synthase expression level reduction and benefited clinically in their lung blood vessel usually of pulmonary hypertension from sucking nitric oxide.Therefore the present invention especially can be used for treating pulmonary hypertension.Show that also hypoxia causes the active inhibition of eNOS.Therefore the present invention can be used for treating the experimenter of the disease of suffering from hypoxia inducible.Astoundingly, also find ID brain injury after the HMG-CoA reductase inhibitor can be used for reducing apoplexy.

Experimenter's disease feature is that the eNOS of hypoxia inducible is active unusual low-level.In other embodiments, the experimenter suffers from a kind of disease, and it comprises that the eNOS of chemical induction is active unusual low-level.In other embodiments, the experimenter suffers from a kind of disease, and it comprises eNOS unusual low-level of cytokine induction.In some important embodiment, the experimenter suffers from the danger of the hypertensive unusual rising of pulmonary's hypertension or pulmonary.In other important embodiment, the experimenter has experienced cerebral infarction or has had the danger of the unusual rising of cerebral infarction.In other embodiments, the experimenter suffers from heart failure or carrying out property nephropathy.In other important embodiment, the experimenter by long term exposure in hypoxia condition.

In other important embodiment, the experimenter has experienced the thrombosis incident or has had thrombotic unusual high danger.In other embodiments, the experimenter has the dangerous of atherosclerotic unusual rising or suffers from atherosclerosis.In other important embodiment, the experimenter has the danger of the unusual rising of suffering from impatient infarction or has experienced impatient infarction.In an embodiment again, the experimenter has repeatedly the danger of the unusual rising of perfusion injury.In preferred embodiments, the experimenter of danger who has repeatedly the unusual rising of perfusion injury is organ transplantation receptor (for example, the heart, kidney, liver, etc.).In other important embodiment, the experimenter suffers from homocystinuria.In some other important embodiment, the experimenter suffers from autosomal dominant inheritance, AD cerebral arteries disease (CADASIL) syndrome of infarction and leukoencephalopathy under the cortex.In other important embodiments, the experimenter suffers from neural degenerative imbalance.In preferred embodiments, the experimenter who suffers from the degenerative imbalance of cental system suffers from Alzheimer.

In certain other embodiments, when the unusual rising that need have cerebral infarction according to the experimenter of treatment of the present invention dangerous, the HMG-CoA reductase inhibitor is excluded outside the treatment as these experimenters.

In other embodiments, method and composition of the present invention (for example, L-arginine slow releasing preparation, L-arginine food stick, or the like) can be used for treatment or prevent Alzheimer.In an embodiment again, method and composition of the present invention can be used for treatment or prevention intermittent claudication.In an embodiment again, preparation of the present invention and compositions can be used for increasing vasodilation.

In preferred embodiments, method of the present invention can be used for reducing the cholesterol levels among the experimenter.Experimenter's administration of HMG-CoA reductase inhibitor and L-arginine be can be used for hypercholesterolemia reducing.In one embodiment, this method makes T-CHOL reduce about 50 to about 150mg/dL.In another embodiment, this method makes T-CHOL reduce about 80 to about 100mg/dL.In addition, experimenter's administration of HMG-CoA reductase inhibitor and L-arginine be can be used for reducing low density lipoprotein, LDL (LDL) cholesterol.In one embodiment, this method makes the LDL cholesterol reduce about 40 to about 110mg/dL.In another embodiment, this method makes the LDL cholesterol reduce about 60 to about 100mg/dL.Method of the present invention also can be used for increasing experimenter's middle-high density lipoprotein (HDL) cholesterol.In addition, administration of HMG-CoA reductase inhibitor and L-arginine can reduce the triglyceride among the experimenter.In one embodiment, method of the present invention makes among the experimenter triglyceride reduce about 30 to about 100mg/dL.In another embodiment, method of the present invention makes among the experimenter triglyceride reduce about 45 to about 75mg/dL.

Arginic being applied in jointly among the reduction experimenter of HMG-CoA reductase inhibitor and L-has synergy in the cholesterol levels.Method and composition of the present invention is compared with compositions with other known methods and is shown with wonderful and significant amount cholesterol reducing level.Particularly, HMG-CoA reductase inhibitor and reduced triglyceride and LDL level in significant mode according to using jointly of sustained release l-arginine of the present invention than existing method.In addition, arginic the using jointly than existing method of HMG-CoA reductase inhibitor and slow release L-increases HDL in significant mode.In one embodiment, HMG-CoA reductase inhibitor and L-arginic use jointly to compare with administration of HMG-CoA reductase inhibitor only make that T-CHOL reduces about 5% to about 15%.In another embodiment, HMG-CoA reductase inhibitor and L-arginic use jointly to compare with administration of HMG-CoA reductase inhibitor only make that T-CHOL reduces about 5 to about 20mg/dL.In an embodiment again, HMG-CoA reductase inhibitor and L-be arginic to be used jointly to compare with administration of HMG-CoA reductase inhibitor only and makes that total LDL cholesterol reduces about 2 to about 20mg/dL more.In an embodiment again, HMG-CoA reductase inhibitor and L-be arginic to be used jointly to compare with administration of HMG-CoA reductase inhibitor only and makes that triglyceride reduces about 5 to about 50mg/dL more, perhaps alternatively about 20 arrives about 35mg/dL.

In another embodiment, method of the present invention can be used for reducing the proteins C reactive among the experimenter.Proteins C reactive is that health is replied the acute phase reactant that acute injury, infection or other inflammatory stimulus discharge.The positive correlation between proteins C reactive and the coronary artery disease has been illustrated in research.Ridker, Circulation 108 (12): e81-85 (2003); People such as Blake, Am.J.Physiol.Regul.Integr.Comp.Physiol.285 (5): R1250-1252 (2003).In one embodiment, this method makes proteins C reactive reduce about 10% to about 50%, perhaps about 25% to about 35%.

Arginic being applied in jointly in the reduction proteins C reactive of HMG-CoA reductase inhibitor and L-has synergy.In one embodiment, this method is not used the arginic slow releasing preparation of L-and is compared and make that proteins C reactive reduces about 50% to about 90%, perhaps about 65% to about 75% more with administration of HMG-CoA reductase inhibitor.In another embodiment, this method with using the arginic slow releasing preparation of L-not administration of HMG-CoA reductase inhibitor compare and make that proteins C reactive reduces about 80% to about 120%, perhaps about 95% to about 105% more.

In addition, method of the present invention can be used for increasing among the experimenter of the asymmetric diethylarginine (ADMA) with rising nitric oxide and produces and/or increase vasodilation.Asymmetric diethylarginine (ADMA) is endogenous, the competitive inhibitor of eNOS.The existence of the plasma ADM A level that raises is relevant unusually with endothelial function.Statins stimulates the expression of external endothelial NO synthase (eNOS) and strengthens the vasodilation dependence endothelium, the NO-mediation in the body.Therefore, statins (for example, simvastatin) can strengthen the endothelial function among the patient of the ADMA with rising.Do not wish to be bound by theory, think that the L-arginine has overcome the inhibition effect of ADMA.

By the experimenter to ADMA with rising use the L-arginine and, randomly, HMG-CoA reductase inhibitor (for example, simvastatin), method of the present invention can be used for increasing nitric oxide and produces and/or increase vasodilation.This using jointly can make endothelial function increase about 5% to about 15% or alternatively, about 7% to about 12%.According to one embodiment of the invention, it is unusual that the experimenter has endothelial function.

For arbitrary mode of administration, the actual amount of the chemical compound of being sent, and realize that the favourable required dosage regimen of pharmacokinetics collection of illustrative plates described herein will depend in part on such as the factor of the bioavailability of this chemical compound (and/or its active metabolite), the imbalance of being treated, desirable therapeutic dose and those skilled in the art conspicuous other factors.The technical staff need not too much test by the blood plasma level that monitors the chemical compound used and/or its active metabolite and regulate and realize that dosage that desirable pharmacokinetics collection of illustrative plates is required or dosage regimen can easily determine the actual amount and the dosage regimen of sending.

Use various route of administration or pattern, can will send in the experimenter so that avoid or reduce as describing the preparation be used for the inventive method or its pharmaceutically acceptable addition salt or hydrate herein according to undesirable side effect of the present invention.In one embodiment, the experimenter is an animal.In another embodiment, the experimenter is a mammal.In a further embodiment, the experimenter is the people.Any character and seriousness that will depend on the disease of being treated to optimum approach in the stable condition.Preferred route of administration of the present invention is the per os approach.These compositionss can provide with unit dosage forms easily, and the either party's method preparation by knowing in the pharmaceutics field.Technology and the prescription of using compositions can be at Remington:the Science ﹠amp; Practice of Pharmacy, Alfonso R.Gennaro, the 20th edition, Williams﹠amp; Wilkins finds in 2000.

The amount of the purpose that preparation of the present invention will be usually planned with effective realization is used, for example, and to treat and/or prevent cerebrovascular and/or cardiovascular disease or imbalance." treatment effective dose " refer to effectively treat disease, imbalance, with disease or the relevant symptom of lacking of proper care, the perhaps amount of the tendency of disease or imbalance.As described previously, term " treatment " refers to the patient is used or administering therapeutic agent or preparation, perhaps to from the isolating organizations of patient or administering therapeutic agent or preparation, wherein this patient suffers from disease, imbalance, with the disease or the relevant symptom of lacking of proper care, the perhaps tendency of disease or imbalance, purpose is to cure, treatment, alleviate, remove, change, remedy, prevention, improve, postpone the outbreak of disease or imbalance and/or incident, delay the deterioration of disease or imbalance, improve or influence this disease or imbalance, the symptom of disease or imbalance, the perhaps tendency of disease or imbalance and/or incident.Determine that the treatment effective dose will be within those skilled in the art's ability, particularly with reference to provided herein open in detail after.

Be suitable for use in pharmaceutical preparation of the present invention and comprise such preparation, wherein contain the L-arginine and/or the HMG-CoA reductase inhibitor of treatment effective dose (amount of the purpose that promptly effective realization is planned) in the said preparation.Usually, effective dose is to produce the amount of desirable pharmaceutical preparation of replying separately or with other preparations.This can comprise the deterioration that only temporarily delays disease.In another embodiment, it comprises the deterioration that forever stops this disease or postpones the outbreak of this disease or disease or prevent its generation.Can monitor the effect of dosage by conventional method to arbitrary disease specific.Certainly, this tittle will depend on the concrete disease of being treated, the seriousness of this disease, the parameter of individual patient, comprise the persistent period of age, physiological condition, size and body weight, treatment, the character of concurrent treatment (if there is), the concrete approach of using and health approach personnel's knowledge and the similar factor in the know-how.

Usually, the dosage of reactive compound will for every day about 0.01mg/kg to about 1000mg/kg every day.In one embodiment, expect that about dosage of 50 to about 500mg/kg will suit.In another embodiment, using is to use per os and every day once or several.

Certainly, the actual amount of L-arginine and/or HMG-CoA reductase inhibitor will depend on experimenter's situation, experimenter's body weight and metabolism.For example, when being applied to the experimenter who suffers from IC or AD, the L-arginine that tablet, pill, dragee, capsule, gelcap, lozenge or capsule contained and/or the amount of HMG-CoA reductase inhibitor will effectively alleviate the illeffects of blood flow deficiency to normal structure, promptly, prevent the experimenter that treated symptom generation or alleviate existing symptom, perhaps prolong the time-to-live.Determining of effective dose is within those skilled in the art's the limit of power, particularly with reference to describe in detail herein open after.

Can also estimate to be used for the treatment effective dose of philtrum from animal model.For example, can prepare the dosage that is used for the people finds in the effective concentration of animal with realization.

Can also estimate the treatment effective dose from people's pharmacokinetic data.Yet do not wish to be fettered by any concrete theory, think that usefulness and experimenter are relevant to total exposure of the application dose of the medicine used and/or its active metabolite, this can determine by the area of measuring under blood concentration-time graph (AUC).Thereby a certain dosage used of expection the method according to this invention is effectively, the AUC of the chemical compound of being used that this dosage had (and/or its active metabolite) be known AUC to the effective dosage of indication about 50% in.Preferred dose be the AUC of the chemical compound (and/or its active metabolite) used be known effective dose AUC about 70%, about 80% or even about 90% and above scope in.By the standard pharmaceutical procedures in cell culture and the laboratory animal, for example, the method that is used for definite LD50 (the lethal dosage of 50% colony) and ED50 (the effective dosage of 50% mass treatment) can be determined the toxicity and the therapeutic efficiency of these reagent.Dose ratio between toxicity and the therapeutic effect is therapeutic index and can be expressed as ratio LD50/ED50.Preferably demonstrate the exponential preparation of big treatment.Although can use the preparation that demonstrates toxic side effects, but should note designing a kind of delivery system, this delivery system with the position of the fixed tissue of being attacked of these preparation targets in case make to the cell of uninfection may damage minimum, thereby, reduce side effect.

Can be used for preparing a series of dosage that are used for the people from cell culture mensuration and zooscopy gained data.In one embodiment, the dosage of these preparations of the present invention has very little or does not have toxicity in comprising the circulation composition scope of ED50.Dosage can change in this scope according to used dosage form and the route of administration of being utilized.For the arbitrary preparation that is used in treatment of the present invention or the prevention method, can measure from cell culture and treat effective dose according to a preliminary estimate.Can be in animal model ruleization dosage, this dosage has been realized the circulating plasma concentration range, it comprises as the IC50 that determines in cell culture (that is, realizing the concentration of the half maximum test-compound that suppresses of symptom).This information can be used for determining more accurately the useful dosage of philtrum.By for example, high performance liquid chroma-tography can be determined the level in the blood plasma.

Based on said method, especially be adjusted in and realize among the experimenter that most powerful dosage is within the those of ordinary skill limit of power based on the haemoconcentration of institute's administered compound and persistent period and/or its active metabolite.

III. production method

What have been found that L-arginine granule in the substrate effectively and in a large number mixes or covers the sustained releasing character of having improved compositions of the present invention.For cellulose matrix, when contacting with water, this substrate is formed gel layer by the part aquation, the arginic rate of release of its control L-.Effective parcel of L-arginine granule or mix and produced temporary transient dissolving barrier, it has postponed, and L-is arginic to be sent.A large amount of slits make that the dissolving of L-arginine is too fast in the substrate.Method of the present invention causes comparing the product with improved properties with the product that produces by direct compression.In addition, method of the present invention is favourable with respect to comprising that the dispersive method of fluidisation is compared, because these methods are time-consuming and expensive.

Effectively and the key that covers efficiently be to implement granulation of the present invention, mill and blend step in.With reference to figure 5; in preferred embodiments; produce tablet according to a kind of method; this method comprises granulation L-arginine (step 110), the L-arginine (step 125 of milling; 140), the L-arginine is mixed (step 145,150,155) and composition compression formed tablet (step 160) with remaining composition.Preferably, this method also comprise the screening composition (step 105) and/or during the step of milling dry L-arginine (step 135) step one the step or two steps.

If screen composition (step 105) before use, #20 and/or #30 mesh sieve can be used for some or all compositions.In preferred embodiments, in granulation (step 105), and before the (not shown) of milling, screen granule once more.The granule that screening provides has narrower particle size distribution, and this is favourable for coating and/or compression.

Granulation step is favourable, because it provides more homogeneous granules.Use arbitrary suitable method as known in the art to make granular activating agent or granulation.Spheroidizing or granulation are generally defined as the process that size increases, and its small particles is gathered into bigger, persistent aggregation, and wherein original granule still can be identified and make them to become flowable state freely.Before the granulation, bonding agent can be added activating agent to improve granulation.During granulation, can add other additives.These additives comprise, for example, and sweetener, flavoring agent, coloring agent, antioxidant, or the like.

Randomly, can add entry or other solvents to help granulation.The water that is added or the amount of solvent depend on, for example, the selection of granulating method, and those of skill in the art can easily determine.Can during granulation, add entry or other solvents by arbitrary suitable time point.For example, bonding agent can mix to form granulating agent with solvent (for example, water), and granulating agent can be sprayed on the activating agent then.Alternatively, if granulating agent thickness and can not being sprayed to equably on the activating agent too wishes bonding agent is at first mixed with activating agent so, spray water or other solvents are to produce the homogeneous form of activating agent granule or granule then.

Arbitrary suitable granulating method all can be used for producing the granule that contains activating agent.Can use moistening granulation and/or dry granulation method.

Dry granulation refers to the granulation preparation and does not use heat and solvent.The dry granulation technology generally includes slugging or the compression of rolling.Slugging is the dry mixed preparation and said preparation is compressed into bolus or piece agent (slug) on compressor.Gained tablet or piece agent are milled and produce granule.The cylinder compression is similar to slugging, but in the cylinder compression, uses the cylinder compressor to replace pelleter.See, for example, Handbook of Pharmaceutical Granulation Technology, D.M.Parikh editor, Marcel-Dekker, Inc.102-103 page or leaf (1997).The dry granulation technology can be used in some situation, for example, and when activating agent is responsive to heat or solvent.

Alternatively, can use moistening granulation.In moistening granulation, solvent and bonding agent are added into preparation usually so that the bigger gathering of granule to be provided.Temperature during the granulation can be set at arbitrary suitable point, is no more than the fusing point of arbitrary component of said preparation usually.Usually, with this mixture about 35 ℃ under about 65 ℃ temperature granulation less than about 20 minutes, more preferably at room temperature about 1 to about 10 minutes (seeing embodiment 8).Usually with granule suitable time of air drying (for example, one or several hours).

Preferably, by high-shear mixer granulation (" HSG ") or fluid bed granulation (" FBG ") granulation activating agent.These two kinds of granulating methods all provide bigger granule or granule, but they are different on the mechanism of used equipment and this method operation.Use can be implemented these granulation technique by the available device of commercial sources.

In HSG, finish mixing and moistening massing by the high mechanical agitation of impeller or chipper.Realize mixing, the multiviscosisty of moistening material by shearing force that impeller produces and compression stress, and reunite.The major function of chipper is the distribution of piece being cut into littler fragment and helping the liquid bonding agent.The liquid bonding agent is imported into roller or is sprayed on the powder to realize more uniform liquid distribution.

On the other hand, fluidisation is such operation, by the tiny solid of this operation by contact the similar liquids state that is converted to other.Under certain gas velocity, this liquid will be supported these granules, make their freely-movables and not be pulled away.The similar violent ebullient liquid of this fluid bed, the motion that the solid particle experience is very violent, this motion is along with gas velocity increases.Thereby fluid bed granulation is a kind of like this method, by this method, by fluid bed the bonding agent solution spray is formed bigger granule to the fluidized powder bed and has produced granule.Bonding agent solution can be from for example, the spray gun ejection of settling in any suitable mode (for example, top or bottom).Spray position and spray velocity depend on the character of used activating agent and bonding agent, and can easily determine by those skilled in the art.

In according to the preferred method of the present invention, granulation L-arginine (step 110) comprises the L-arginine mixed with bonding agent such as polyvinyl pyrrolidone and forms mixture (step 115), and with mixture and granulating agent (granulation carrier) granulation (step 120) in granulator.Granulating agent can be for example, to be dissolved in the polyvinyl pyrrolidone in the purified water.Preferably, use high shear granulator, as Niro PMA65 high shear granulator.This granulator can be used for mixing L-arginine and bonding agent, after also can be used for being sprayed to the granulation carrier on the mixture with this mixture granulation.

After one or more components of granulation preparation, randomly, the preparation of the granulation of can milling.Can implement to mill with arbitrary suitable available equipment of commercial sources (for example, the CoMil of 0.039 inch sieve of equipment) that passes through.Can select the width of mesh of sieve according to the size of desirable granule.After the activating agent of granulation milled, can be if wish with they further dry (for example, in air).

In preferred embodiments, the L-arginine of milling comprises according to the technology of knowing in this area (sees U.S. Patent number 5 usually, 145,684 and european patent application 498,482, the content of these two applications all is merged in this paper as a reference) the moistening granule of milling or moistening mill (step 125), dry granule (step 130) and mill dry granule or drying mill (step 140).Grinding machine such as CoMil can be used to moistening milling and the drying granule of milling.In one embodiment, grinding machine is used for moistening milling by outfit ' 375Q sieve and be used for drying mills ' 062R sieve.By with granule at bed dryer, for example, be dried to desirable drying loss (LOD) level in the Aeromatic S-2 fluidized bed dryer, for example≤3%LOD, can realize drying steps.(step 135) implements drying steps up to reaching desirable LOD stage by stage.

The L-arginine mixed with remaining composition can comprise pre-blend step (step 145), blend step (step 150) and final blend step (step 155).Pre-blend step can comprise that with L-arginine/polyvinyl pyrrolidone granule and filler and fluidizer for example, microcrystalline Cellulose and colloidal silica mix.For example, in 8 quarts of V-arrangement mixers, mix and to realize pre-blend step in about 5 minutes with 25rpm.Blend step can comprise to this mixture and add one or more slow releasing agents, for example, and one or more hydroxypropyl emthylcelluloses and a kind of filler, for example, microcrystalline Cellulose.Can be in 2 cubic feet of V-arrangement mixers for example mix and to realize blend step in about 20 minutes with 25rpm.Final blend step can comprise that the mixture in 2 cubic feet of V-arrangement mixers adds releasing agent/lubricant, and for example, magnesium stearate was also mixed about 5 minutes with 25rpm.

After having prepared, said preparation compression (step 160) is become tablet form as above-mentioned preparation.By arbitrary suitable method, with or can implement this tablet without compression stress and be shaped.For example; can with arbitrary tablet machine (for example, equipment 0.748 " * 0.380 " oval, projection, the Manesty Beta Press of common die) finish the compression of preparation after the granulation step, preferably; lubricated dose of (for example, magnesium stearate) sufficient lubrication of said preparation compositions.Can utilize the many alternative approach that realize this step, and the present invention is not subjected to the restriction of the use of arbitrary concrete device.Can implement compression step with rotary-type tablet machine.This rotary-type tablet machine has swivel plate, and the latter has a plurality of hole, the perhaps moulds of passing through that are used to form tablet again.Preparation is inserted into this mould also subsequently by pressing mold.

Alternatively, can prepare tablet by molding.In suitable machine, will can prepare the tablet of molding with the moistening pulverulent mixture molding of inert liquid diluent.

The diameter of tablet and shape depend on that selection is used for mould, mold and the punch press of the shaping or the compression of granule composition.Tablet can be dish type, oval, ellipse, circle, cylindrical, triangle, or the like.The tablet scratch can be broken with promotion.Upper surface or lower surface can or be recessed into symbol or alphabetical embossment.

Can be based on the type/model of tablet machine, to desirable those physical propertys of tablet product (for example, desirable hardness, fragility, or the like), desirable tablet appearance and size, or the like, compression stress can be selected.Usually, applied compression power makes the tablet of this compression have the hardness at least about 2kp.These tablets enough hardness and intensity are provided usually and can be packaged, transportation or handled by user.If wish, can be to the higher compression stress of tablet applications to increase tablet hardness.Yet the preferred compression stress of so selecting makes it can not make the granular degeneration that contains activating agent in the tablet (for example, chap or break).Preferably, so applied compression power makes replying less than about 10kp of compressed tablets.In certain embodiments, preferably tablet is compressed to about 3kp to about 7kp, optional about 3kp is to about 5kp, the perhaps hardness of about 3kp.

Usually, the weight of final tablet arrives about 2000mg for about 50mg, and more generally about 200mg is to about 1000mg, and perhaps about 400mg is to about 700mg.

Produce the concrete preparation and the method for the present composition and given unique advantage the sustained release l-arginine compositions.Particularly, preparation of the present invention and method make compositions have desirable sustained-release dissolution scattergram.Randomly, sustained release l-arginine formulations will continue vitro drug release and reach 14 hours at least, discharge about 10% to about 40% in the time of preferred about 1 hour, discharge about 30% to about 70% in the time of about 4 hours, discharge about 55% to about 75% in the time of about 6 hours, discharge about 65% in the time of about 8 hours to about 85%, discharge about 75% in the time of about 12 hours, discharge about 80% to about 100% in the time of about 14 hours to about 95%.As by as illustrated in Fig. 7, preparation of the present invention has been realized this preferred dissolution.In addition, as shown in embodiment 8 and embodiment 14, the dissolving and stability study show preparation of the present invention after production 1 and demonstrated in 2 months the best dissolution profiles figure.

In addition, preparation of the present invention and method make the sustained release l-arginine compositions excessively not frangible.In addition, preparation of the present invention and method make that the sustained release l-arginine compositions is enough compressible to allow convenient preparation said composition.

If wish the embodiment that other modifications can be mixed tablet.For example, by arbitrary technique known, such as, for example, use various coatings, for example, for example with, the ion exchange complex of Amberlite IRP-69 for example can realize that activating agent passes the modification that tablet matrix of the present invention discharges.Tablet of the present invention can also comprise GI move-weaken medicine or use jointly with this medicine.Can also come modification active to produce prodrug by the chemical modification of bioactive compound, this prodrug will be in vivo by release of active compounds such as enzyme or hydrolysis cuttings.Extra layer or coating can be used as diffusion barrier so that the additional method of control drug release speed and time to be provided.

If comprise HMG-CoA reductase inhibitor (for example, simvastatin) and/or extra reagent, these reagent preferably add in blend step (step 145,150,155) so.When tablet contained sustained release l-arginine formulations and HMG-CoA reductase inhibitor preparation, this tablet can have the sustained release l-arginine formulations core and contain second outer covering or coating of the preparation of at least a HMG-CoA reductase inhibitor.Alternatively, tablet can contain the L-arginine formulations, for example, and the HMG-CoA reductase inhibitor preparation on a sustained release l-arginine formulations and a shared surface.

When L-arginine and HMG-CoA reductase inhibitor sequentially or are jointly used, every kind of tablet, cachet, lozenge, or capsule can contain the 0.01mg that has an appointment to about 200mg HMG-CoA reductase inhibitor.The amount of HMG-CoA reductase inhibitor will depend on used concrete HMG-CoA reductase inhibitor.

In another aspect of this invention, the form with food provides the compositions that is used for the treatment of cardiovascular and/or cerebrovascular disease.Preferably, this food is the form of rod type food as the prescription health bar.Utilize food to make that the arginic amount of L-that can provide is bigger than the amount that can mix single tablet, for example, in single tablet, be difficult to mix the L-arginine more than the 1g.Thereby, need a plurality of tablets to send arginic amount above the L-of 1g.The invention provides rod type food, it can provide the above L-arginine of 1g and other reagent (if hope).In one embodiment, the L-arginine is as immediate release formulation, and for example, the arginic granule that discharges immediately of L-is added in the food stick.Preferably, this rod type food comprises slow releasing preparation, and it comprises, for example, and the arginic slow release granule of L-.In preferred embodiments, granule comprises the taste masked composition, for example, and the taste masked coating.In another embodiment, this rod type food also contains extra reagent, as the HMG-CoA reductase inhibitor.Preferably, the HMG-CoA reductase inhibitor is a statins, as simvastatin.L-arginine and statins made up in food carrier will be provided restraining and be easy to use said preparation.When hope more during high dose, use food can also reduce to taking the needs of the arginic a plurality of tablets of L-.

In one embodiment, rod type food has about 1 and arrives about 10g L-arginine to about 80g simvastatin and about 1.In preferred embodiments, provide rod type food, it has altogether at least about 10mg simvastatin and each rod type food about 4g L-arginine or its salt, and sugar, fruit ingredient, protein and vitamin and mineral.The weight of this rod type food is about 25 to about 100g.In concrete grammar,, this syrup is mixed with rod type food in lower temperature and minor constituent by sugar and fruit are stuck with paste in higher temperature mixing.Minor constituent with after syrup mixes, is added L-arginine and simvastatin, especially also add the protein extender, expand then, also add the food agent, especially fruit slice or other tiny edible compositions also add soybean protein so that desirable quality and local flavor to be provided.Products obtained therefrom can be stablized preservation, has the desirable character that influences sense organ aspect good to eat, and provides composition and simvastatin and L-arginine bonded healthy combination.At U.S. Patent number 6,063, method and the prescription of producing the healthy rod type food that contains L-arginine and L-lysine have been described in 432, this patent is incorporated into this paper as a reference by complete.

Another aspect of the present invention is the method for producing above-mentioned rod type food.This method will comprise as above-mentioned and Fig. 5, the granulation L-arginine that step 110 is relevant.Preferably, granulation step will comprise pre-blend step (step 115) and granulation step (step 120).Preferably, this method also comprises the above-mentioned moistening step (step 125) of milling.Moistening granulation by L-arginine and suitable as described above excipient can obtain this rod type food.The gained granule will directly be used or it will be used the taste masked coated cellulose.

Further illustrate the present invention by the following examples, these embodiment should not be understood that restriction.The content of the patent application of all lists of references, patent and the publication of using by the application all is impregnated in this paper as a reference.

Embodiment

Embodiment 1: tablet formulation 1

Will about 250g L-arginine place blender and when it slowly mixed with 100RPM, (Rohm America, Piscataway was NJ) to form wet mass to add 100g EUDRAGIT RS 30D hypotonicity metering system waterborne polymeric dispersant.Make this wet mass pass the 18-20 mesh sieve and allow 50 ℃ of dryings 24 hours.(The Dow ChemicalCompany, Danbury CT) form mixture with 3g magnesium stearate dry mixed with 84g METHOCEL K100M CR methylcellulose with the dry L-arginine granule of gained (250g).With 7/16 arch punch press the gained mixture is compressed into tablet.

Embodiment 2: tablet formulation 2

250g L-arginine is placed blender and when it slowly mixes, adds 84gMETHOCEL K100M CR methylcellulose and 3g magnesium stearate.With 7/16 arch punch press the gained mixture is compressed into tablet.

Embodiment 3: capsule preparations 1

250g L-arginine is placed blender and when it slowly mixes, add 100gEUDRAGIT RS 30D hypotonicity metering system waterborne polymeric dispersant to form wet mass.Make this wet mass pass the 18-20 mesh sieve and allow 50 ℃ of dryings 24 hours.The dry L-arginine granule of gained (250g) and 84g METHOCEL K100 M CR methylcellulose and 3g magnesium stearate dry mixed are formed mixture.The gained mixture places 00 gel capsule.

Embodiment 4: capsule preparations 2

250g L-arginine is placed blender and when it slowly mixes, adds 84gMETHOCEL K100 M CR methylcellulose and 3g magnesium stearate.The gained mixture places 00 gel capsule.

Embodiment 5: tablet formulation 3

With KitchenAid  blender 250g L-arginine is mixed with 50g METHOCEL K100M CR methylcellulose and homogenate; The mixer mixed on low speed formed drying composite in 10 minutes.It is moistening equably up to this material quilt that 115g EUDRAGIT RS 30D hypotonicity metering system waterborne polymeric dispersant is increased progressively the adding drying composite with 5g.This moistening material passes 12 mesh sieves, and 20 mesh sieves were 1% up to moisture in 24 hours 30 ℃ of dryings subsequently by weight then.With dry L-arginine granule of gained and 7g magnesium stearate dry mixed, use Beta Manesy press then, use 7/16 arch punch press to be compressed into tablet.

Embodiment 6: the production of slow releasing tablet

About 1000g L-arginine was mixed about 5 minutes with 100 RPM in GP-1 high shear mixer (granulator) with about 200g METHOCEL K100 M CR methylcellulose.Add about 138g EUDRAGIT RS 30D hypotonicity metering system waterborne polymeric dispersant then, impeller moves with 200RPM, and pressure is 1.5 crust.With 200RPM granulation mixture 1 minute.Then granule is dried to about 2% moisture with 45 ℃ of inlet temperatures and 100 CMH air flows in the MP-1 fluidised bed granulator.Then with the circular impeller mill dry granule of Comil 197S with 55R sieve and 90% speed.In 8Qt.V shape mixer, add about 27g magnesium stearate and mixed 2 minutes to the granule of milling." standard arch mould is compressed into tablet with material, and the tablet weight of this tablet is 682.5mg, has the highest possible hardness with 7/16 with Beta Manesty Press then.With tablet with in every bottle of 60 Manual Packaging and the 75cc HDPE bottle.

Produced this tablet with respect to from BioEnergy (Warren, NJ) the release collection of illustrative plates that passes through the available sustained release l-arginine tablet of commercial sources of Gou Maiing with high performance liquid chromatography (HPLC).Fig. 7 is a chart of describing the release collection of illustrative plates of two kinds of preparations.

The arginic pharmacokinetics assessment of embodiment 7:L-

The adult volunteer who is in 14 health under the fasted conditions has been implemented four-way intersection (four-way crossover) research at random, and this research is used to assess the pharmacokinetics of L-arginine slow releasing tablet to immediate release capsule." health " refers to not have the non-hypercholesterolemia experimenter of cardiovascular risk factors as used herein.The sustained release l-arginine tablet (L-arginine SR) of embodiment 6 has been compared in this research and (Los Angeles, that CA) buys passes through the available L-arginine capsule (L-arginase i R) that discharges immediately of commercial sources from Montiff.

Research purpose is to determine the pharmacokinetic parameter of sustained release l-arginine.As describing in the following table 1, based on the p value of the paired t-check of two tails that every kind of pharmacokinetic parameter is carried out, C _MaxAnd T _MaxBetween statistically-significant difference is arranged.As expected, compare with immediate release capsule, the sustained release l-arginine tablet has lower C _Max(14.9 μ g/mL are to 24.1 μ g/mL) and longer T _Max(4.4h is to 1.4h).

Table 1:L-arginine SR is to the PK parameter of L-arginase i R

The L-arginine	C max	AUC 0-t	AUC 0-10	T max0-t	T max0-10
						L-Arg?SR	14.9	143	68.56	4.4	3.27
L-Arg?IR	24.1	147	92.23	1.4	1.35
						The % ratio	0.62	0.97	0.74	3.2	2.43
The P value	0.0005	0.677	0.0382	0.0133	0.0073

Embodiment 8: produce improved sustained release l-arginine tablet

Table II has been listed the composition that is used to produce improved slow releasing tablet, and every kind of amount that composition is used.

Table II: composition

Component	The mg/ tablet	Percent (%)	Weight/crowd (Kg)
				L-arginine mono-hydrochloric salts	500	50	12.5
Polyvinyl pyrrolidone (K 29/32)	35	3.5	0.88
				Purified water	-	-	2*
Hydroxypropyl emthylcellulose (METHOCEL K100M P CR)	275	27.5	6.87
				Hydroxypropyl emthylcellulose (METHOCEL E 4M CR)	75	7.5	1.88
Microcrystalline Cellulose (AVICEL PH 102)	102.5	10.2	2.56
				Colloidal silica	5	0.5	0.13
Magnesium stearate	7.5	0.75	0.18
				Total:	1000	100.0	25

* make water in the granulation, then drying composite

Except magnesium stearate, all the components all screens in the #20 mesh sieve.Magnesium stearate is screened in the #30 mesh sieve.To make an appointment with half polyvinyl pyrrolidone (polyvinylpyrrolidone) to be dissolved in purified water and set aside as granulating agent.With L-arginine and remaining polyvinyl pyrrolidone dry mixed 4 minutes in Niro PMA 65 high shear granulator, then by to about 6.5 minutes of mixture spraying granulating agent with this mixture granulation.Then at equipment ' the moistening granule of milling among the CoMil of 375Q sieve.In Aeromatic S-2 fluid bed dryer, the granule of milling is dried to then＜3% LOD.Then at equipment ' mill dry granule among the CoMil of 062R sieve.In 8 quarts of V-arrangement mixers, will make an appointment with microcrystalline Cellulose of half and colloidal silica to mix 5 minutes and transfer in 2 cubic feet of V-arrangement mixers then with 25rpm.The remainder of microcrystalline Cellulose and hydroxypropyl emthylcellulose also are added in 2 cubic feet of V-arrangement mixers and with 25rpm and mixed 20 minutes.In 2 cubic feet of V-arrangement mixers, add magnesium stearate then and mixed 5 minutes with 25rpm.At last, the Manesty Bet Press with the common die of equipping 0.748 " * 0.380 " elliptical shaped lobes is compressed into target weight 1000mg with mixture.Fig. 6 is the indicative flowchart of this method.

Can use controlled trial and specification in the process of standard during production process, the controlled trial and the specification infra Table III that are used for this embodiment are listed.

Table III: contrast in the L-arginine SR tablet process: specification and method

Specification	Method	Accept standard
			Mixing uniformity	CTMLP-663	On average: the 90.0%-110.0% RSD% NMT 5.0% that label is claimed
Volume and tap density	SOP?Lab?2010	The report result
			Particle size distribution	SOP?Lab?2018	The report result
Moisture	SOP?Lab?2059	NMT?3.5％

Can use standard method for releasing and specification, be used for providing in the infra Table IV of this example:

Table IV: L-arginine SR tablet method for releasing and specification

Specification	Method	Accept standard
			Physical appearance	Visual inspection	White arrives canescence tablet ellipse, the tablet of projection
Identify	CTMPLP-663	Corresponding to reference standard, the retention time of sample and online UV spectrum (200-400nm),
			Render a service	CTMLP-663	The 90.0%-110.0% that label is claimed
Related substances	CTMLP-663	Single: NMT 0.5% is total: NMT 2.0%
			Moisture	SOP?LAB?2059	NMT?3.5％
Dissolution profiles figure	CTMLP-663	1hr 10-40% 4hr 30-70% 12 hr 〉=75% record collection of illustrative plates
			Content uniformity	CTMLP-663	USP<905>
The microorganism restriction	USP<61>	Mould and the sum≤50cfu/mL yeast addition of total aerobe number≤100cfu/mL does not have Escherichia coli and does not have S.aureus and do not have P.aeruginosa and do not have salmonella

In addition, desirable physical features has been illustrated in research, comprises the fragility and the content uniformity of sustained release l-arginine formulations of the present invention.

Table V: the physical test of two batches of SR-L arginine formulations, effectiveness, content uniformity and dissolving

Criticize #	1	2
			Average tablet weight n=20mg	1003.3	1014.5
Tablet hardness (n=20) kp	11.0	12.4
			Tablet thickness (n=20) mm	7.89	7.70
Tablet fragility %	0.1	0.1
			Render a service %	98.4	100.5
Content uniformity n=10; %	99.0	100.8
			Content uniformity %RSD	1.5	1.8
Dissolution time hr	% discharges
			?0	0	0
?1	27.3	26.8
			?2	42.1	42.1
?4	59.9	60.2
			?6	73.0	73.6
?8	82.8	83.4
			?10	90.3	90.3
?12	95.1	94.9
			?14	98.4	92.5

Embodiment 9:L-arginine SR with and not with simvastatin and simvastatin with and do not assess with the pharmacokinetics of L-arginine SR

Studied L-arginine SR with and not with simvastatin and simvastatin with not with the pharmacokinetics of L-arginine SR.Used the L-arginine SR tablet of embodiment 6 and passed through commercial sources available from BioEnergy (Warren, NJ) simvastatin tablet of Gou Maiing.

As can be seen, check the p value that obtains based on the paired t of two tails, at C to every kind of pharmacokinetic parameter enforcement from Table VI _Max, AUC _0-10, and T _MaxProcessing between do not have statistically-significant difference.As describing in the Table VII, L-arginine SR does not have the statistics appreciable impact to the single dose kinetics of simvastatin.

Table VI. there is and do not have the L-arginine PK parameter of simvastatin

The L-arginine	C max(mg/ml)	AUC 0-10(mg-hr/ml)	T max(hr)
				L-Arg?SR	14.77	68.56	3.27
L-Arg SR and simvastatin	13.49	51.55	3.23
				The % ratio	1.09	1.33	1.01
The P-value	0.5001	0.0713	0.9716

Table VII. there is and do not have the PK parameter of the arginic simvastatin of L-

Simvastatin	C max(ng/ml)	AUC 0-10 (ng-hr/ml)	T max (hr)	K elim (1/hr)	t 1/2 (hr)
						Simvastatin w/o L-arginine SR	21.15	107.93	2.68	0.1248	6.56
Simvastatin and L-arginine SR	18.95	114.36	2.29	0.0950	10.01
						The P value	0.5360	0.6302	0.4758	0.1526	0.1059

Embodiment 10: the arginic influence of using infarction size in the mice of simvastatin and L-

In mice, studied and used the influence of simvastatin and L-arginine the infarction size.Mice is dissolved in the solution that contains simvastatin in the saline solution and contains simvastatin and the arginic solution of L-by peritoneal injection, and consumption as shown in Figure 3.Described the infarction size result in these mices and the matched group among Fig. 2 and Fig. 3.

Embodiment 11: the injectivity optimizing of simvastatin and the combination of L-arginine

In mice, studied the injectivity optimizing of simvastatin and the arginic combined administration of L-.Mice is injected the simvastatin and the L-arginine of varying level, as shown in Figure 4.The result of this research also shows in Fig. 4.Statistical analysis predicts that the optimum range of this combination is 1.2-1.4mg/kg simvastatin and about 20-25mg/kg L-arginine.

Embodiment 12: by uniting the vasodilative improvement that relies on endothelium among the patient who strengthens the rising of ADMA level with L-arginine slow release

Statins stimulates the expression of external endothelial NO synthase (eNOS) and strengthens the vasodilation of the NO mediation that relies on endothelium in the body.Asymmetric diethylarginine (ADMA) is a kind of endogenous competitive inhibitor of eNOS.The rising of plasma ADM A level is relevant unusually with endothelial function.Only find that when overcoming the inhibitory action of ADMA simvastatin just strengthens the endothelial function among the patient of ADMA rising by the L-arginine slow release that replenishes.

In the cross-over design in three cycles, the old experimenter that 15 clinical asymptomatic ADMA levels raise accepts simvastatin (40mg/ days), L-arginine slow release (3g/ days) or their combination as describing among the embodiment 8 with random order, use lasting 3 weeks for every kind, have removing (wash-out) time in three weeks between the treatment at least.Rely on the vasodilation of endothelium by the ultrasonic assistant images analysis and evaluation that uses a computer of brachial artery; Determine ADMA and L-arginine blood plasma concentration by effective HPLC method.

To 15 patients' finishing research analysis disclose with treatment before measure and compare, sustained release l-arginine separately or with the vasodilation percentage ratio of simvastatin associating all enhancing dependence endothelium.Unite and make than only increasing by 3.87% (p＜0.025) with the preceding variation that relies on the vasodilation percentage ratio of endothelium of the observed treatment of simvastatin.Associating and only to rely on the difference of change of vasodilation percentage ratio of endothelium between the sustained release l-arginine very little.The vasodilation of the dependence endothelium that glyceryl trinitrate causes is not subjected to the influence of arbitrary treatment.L-arginine slow release has all significantly improved blood plasma L-arginine/ADMA ratio (being respectively baseline 82.3 ± 4.0vs.102.8 ± 9.2 and 102.6 ± 10.8, every kind of p＜0.05) separately or with the simvastatin associating.These results summarize in Fig. 8.

Simvastatin can not strengthen the endothelial function among the experimenter of the upborne ADMA level of eNOS blocking-up; Simvastatin and uniting of oral L-arginine slow release have synergism to endothelial function.Because the effect of NO-mediation can play a major role in the curative effect of statins, so in the patient that ADMA concentration raises, should consider combination with L-arginine slow release.

Embodiment 13: the therapeutic alliance by simvastatin and L-arginine slow release improves cholesterol levels

In the research of in embodiment 12, describing, analyzed the change of before treating and treatment back T-CHOL (TC), LDL cholesterol, HDL cholesterol and triglyceride.The result of this analysis shows in Fig. 9.As illustrated in the result, the using jointly and only use simvastatin and compare and reduced T-CHOL, LDL cholesterol and triglyceride and increased the HDL cholesterol to a greater degree of sustained release l-arginine of the present invention and simvastatin.

Embodiment 14: the dissolving of determining arginine HCl in the slow release arginine HCl 500mg tablet by HPLC discharges

Be prepared as follows mobile phase.At first, the about 0.9g 1-of weighing sodium pentanesulfonate salt monohydrate and 3.5g biphosphate sodium-hydrate add 1 liter of pH3.3 buffer of preparation in the suitable container.Add about 100mL deionized water dissolving.Regulate pH to 3.3 by adding phosphoric acid.Subsequently, 850mL pH3.3 buffer and 150mL methanol are merged in the suitable container and mix.Mixture filters by 0.45 μ m nylon membrane filter.The degassing before at last mixture being used.

Be prepared as follows dissolve medium (the 50mM phosphate buffer of pH6.8).At first, 20.0mL 10M NaOH is drawn in the 1000mL volumetric flask and with the deionized water dilution with preparation 0.2M NaOH.Subsequently, in suitable container, add the 54.44g anhydrous potassium dihydrogenphosphate, with 2000mL deionized water dissolving and dilution.Add 896mL 0.2M NaOH and be diluted to 8000mL to this container with deionized water.At last, this mixture outgases before use.

Be prepared as follows sample dissolution.The Hexaarginine HCl500mg tablet of the preparation of describing among weighing such as the embodiment 8.Each tablet is placed the rustless steel sinker (sinker) that contains 900mL phosphate buffer (pH6.8).The container that subsequently this sinker is fallen into USP Apparatus 2 (slurry) rotates with 75rpm immediately at 37 ℃ ± 0.5 ℃.Take out 10mL solution for dissolving each time point of analyzing at 1,2,4,6,8,10,12 and 14 hour time point separately from container.Each of these sample solutions is all passed through 0.45 μ m PVDF syringe filter and is filtered.Filtrate collection is used for analyzing in the HPLC bottle, wherein a 1-2mL is abandoned.Use 10 μ mFull Flow Filter, the 10mL dissolve medium that will be preheating to 37 ℃ ± 0.5 ℃ behind each sample point turns back in the dissolution vessel.Operator should be understood that this sample solution at room temperature stablized at most 1 day, stablize at most under 4 ℃ 3 days.

Be prepared as follows arginine HCl standard solution.Accurately weighing 28 ± 2mg arginine HCl reference standard thing places the 50mL volumetric flask.With dissolve medium with standard dissolving and be diluted to volume.

(5 μ m, 250mm * 4.6mm) implement HPLC uses UV to detect at the 210nm place to use BDS Hypersil C18 post.Column temperature is set to room temperature.Usually, be 9 minutes running time, and volume injected is 10 μ L, and flow velocity is 0.8mL/min, and mobile phase is pH3.3.Buffer/methanol (85/15, v/v), as above-mentioned preparation.

Each tests following carrying out.After injecting dissolve medium, continuous then 5 injection arginine HCl standard solution and last injection be every kind of sample solutions once.Inject arginine HCl standard solution when per 6 samples injection back and order end of run once more.System drifting in the whole service (that is, with 5 times of arginine HCl standard solution the recovery percentage ratio of the average comparison with standard solution of injection) continuously should be about 97% to about 103%.

When the arginic percent that determine to discharge, operator should guarantee carefully that the USP at arginine HCl peak in the injection of working standard solution trails the factor (T) less than 2.Following calculating T:

T＝W _.05/2f

W wherein _.05Be the peak width apart from the arginine HCl peak, 5% place of the peak height of baseline, f is the distance (in 5% distance of locating to measure of the peak height of distance baseline) between the forward position at peak maximum and peak.

The arginine HCl percent that following calculating discharges:

Wherein n is the sum of measuring, V _rIt is the volume (10mL) of the dissolve medium of every kind of measurement, V is the initial volume (900mL) of dissolve medium, Cs is the concentration (mg/mL) of arginine HCl in the working standard solution, Ci is the concentration (mg/mL) (wherein i=1 is to i=n-1) of arginine HCl in each sample solution, Ru is for to reply from the peak area at the arginine HCl peak that sample solution obtains, Rs replys for the average peak area at the arginine HCl peak that the continuous injection from working standard solution obtains, and LC is that the label of arginine HCl is claimed (500mg).

Calculate the release percent of locating in 1,2,4,6,8,10,12 and 14 hour.Table VI and VIII have summarized the result of various solution researchs.

Table VIII: the dissolution profiles figure of L-arginine SR tablet under about 40 ℃/75%RH stability

Time point	Initial			1 month 2 months
							Dissolution time, hr	% discharges
0	0	0	0
				1	20.4	21.8	28.1
2	36.4	36.6	41.1
				4	53.5	54.3	58.5
6	66.8	67.5	71.5
				8	76.6	77.9	81.3
10	83.1	85.5	88.3
				12	87.2	89.7	92.9
14	89.1	92.4	96.0

The adjusting of the dependence simvastatin that embodiment 15:eNOS expresses

Below scheme be used for studying the mechanism that the eNOS function relies on the increase of simvastatin, this research uses the human aorta endotheliocyte of cultivating (HAEC) to distinguish in the rise of eNOS function protein synthesis and protein mobilization or protein activation again.

According to following method cultivate human aorta endotheliocyte (HAEC-c) (BioWhittaker, Walkersville, MD).Endothelial cell growth in the EBM-2/EGM-2 culture medium (BioWhittaker) converges to about 90% to about 80%.Wash the cell in each bottle and add the 15ml culture medium with the 5ml culture medium to every kind of cell.Make cell detachment and it is transferred in the 50ml conical pipe with cell scraper.By at centrifugal 8 minutes sedimentation cells of 800RPM.Abandon supernatant also with cold 1 * PBS washing precipitation.

Following homogenate cell.Make precipitation fluff and add 400 μ l, 10 * homogenate buffer (250mM Tris, pH7.4,10mM EDTA and 10mM EGTA).With about 10 times of 27G pin homogenate sample.Homogenate is transferred in the 1.5ml epindorph pipe.Subsequently precipitation is resuspended in 30 to 45 μ l, 1 * homogenate buffer.

Following analysis of cells.(BioRad Laboratories, Hercules CA) prepare paste resin with 5 bed volume 0.5N NaOH washing resin AG 50W-X8 in suitably big or small pillar.With 20 volume water washing pillars.With termination/level pad (the 50mM sodium acetate, pH5.5) the balance resin up to eluate in the 0.05pH unit of termination/level pad.Gained solution as 50% serosity in the termination/level pad 4 ℃ of preservations.In addition, prepare fresh 10mMNADPH (pH7.4) among the 25mM tris by the 5mg bottle that 602 μ ltris is added to the NADPH that weighs in advance.The 0.069mg calmodulin, CaM is added preparation 1 μ M calcium accent solution in the 4.1mL water.Also prepare 8 water-soluble μ M CaCl.By mixing 50mM Tris (pH7.4), 6 μ M BH ₄, 2 μ M flavin adenine dinucleotide (FAD)s and 2 μ M flavin adenine mononucleotides preparation, 2 * reaction buffer.Subsequently, by mixing 25 μ l, 2 * reaction buffer, 5 μ l 10mMNADP, 5 μ l 8mM CaCl ₂, 4 μ l calmodulin, CaM solution and 1 μ l ¹⁴C arginine preparation 2 * reactant mixture.Merge with 40 μ l reactant mixtures and 5 μ l samples or to impinging upon in the 1.5ml centrifuge tube.This centrifuge tube was hatched 1 hour at 37 ℃.

Prepare pillar from 1ml head of pipette excision top with the minimum diameter that increases this head of pipette.The resin serosity that 250 μ l are as above described preparation be pipetted into each pillar (FisherScientific, Glenlake, IL).(the 50mM sodium acetate, pH5.5) the washing pillar is twice with 400 μ l termination/level pads.

After hatching, to each sample and contrast add about 400 μ l termination/level pads (the 50mM sodium acetate, pH5.5).Add 400 these mixture of μ l to equilibrated pillar.Wash each pillar with 400 μ l termination/level pads.400 μ l post eluates are transferred in the scintillation vial that the 4ml scintillation solution is housed.Gained solution fully mixes on vortice.With scintillation counter (Beta counter, Beckman Coulter, Inc., Fullerton CA) obtains desirable counting.Part is passed through from each sample subtracting background (buffer contrast) result of calculation.Sample value is expressed as the percent of count per minute or untreated cell.

L-arginine by labelling is expressed as the percent of the citrulline of untreated cell generation to the relative eNOS function of converted measurement of L-citrulline and with it.Figure 10 has shown experimental data, wherein before definite eNOS function HAEC and 1.0 or 0.3 μ M simvastatin is hatched 24 hours.Cultivate untreated cell simultaneously and these cells are used to calculate relative eNOS function.Figure 10 shows that simvastatin has increased the level of eNOS function in the endothelial cells cultured with removing.

All data show that simvastatin influences eNOS expression and function in the endotheliocyte.The increase that relies on simvastatin in the rise of eNOS function in the special mRNA of the needs of protein synthesis and eNOS and the function is consistent with the drug-induced adjusting model of eNOS genetic transcription.

Equivalence

Those skilled in the art only use normal experiment just can recognize or many equivalence of the particular of the present invention that can determine to describe herein.These equivalence will be comprised by following claims.

Claims (93)

1. reduce the method for cholesterol among the experimenter, this method comprises experimenter's administration of HMG-CoA reductase inhibitor and contains the arginic slow releasing preparation of L-.

2. the process of claim 1 wherein that the HMG-CoA reductase inhibitor contains slow releasing preparation.

3. the process of claim 1 wherein that this method reduces T-CHOL and low density lipoprotein, LDL (LDL) cholesterol among the experimenter.

4. the process of claim 1 wherein that the HMG-CoA reductase inhibitor contains simvastatin.

5. the process of claim 1 wherein that this method makes T-CHOL reduce about 50 to about 150mg/dL.

6. the process of claim 1 wherein that this method makes T-CHOL reduce about 80 to about 100mg/dL.

7. the process of claim 1 wherein that this method makes the LDL cholesterol reduce about 40 to about 110mg/dL.

8. the process of claim 1 wherein that this method makes the LDL cholesterol reduce about 60 to about 100mg/dL.

9. the process of claim 1 wherein that this method increases experimenter's middle-high density lipoprotein (HDL) cholesterol.

10. the process of claim 1 wherein that this method makes triglyceride reduce about 30 to about 100mg/dL.

11. the process of claim 1 wherein that this method makes triglyceride reduce about 45 to about 75mg/dL.

12. the process of claim 1 wherein this method with do not make when experimenter's administration of HMG-CoA reductase inhibitor not being used compare when containing the arginic slow releasing preparation of L-that T-CHOL reduces about 5% to about 15%.

13. the process of claim 1 wherein this method with do not make when experimenter's administration of HMG-CoA reductase inhibitor not being used compare when containing the arginic slow releasing preparation of L-that T-CHOL to be reduced at least about 5 to about 20mg/dL.

14. the process of claim 1 wherein this method with do not make when experimenter's administration of HMG-CoA reductase inhibitor not being used compare when containing the arginic slow releasing preparation of L-that the LDL cholesterol to be reduced at least about 2 to about 20mg/dL.

15. the process of claim 1 wherein this method with do not make when experimenter's administration of HMG-CoA reductase inhibitor not being used compare when containing the arginic slow releasing preparation of L-that triglyceride to be reduced at least about 5 to about 50mg/dL.

16. the process of claim 1 wherein this method with do not make when experimenter's administration of HMG-CoA reductase inhibitor not being used compare when containing L～arginic slow releasing preparation that triglyceride to be reduced at least about 20 to about 35mg/dL.

17. increase the method that nitric oxide produces among the experimenter that asymmetric diethylarginine (ADMA) raises, this method comprises this experimenter's administration of HMG-CoA reductase inhibitor and L-arginine.

18. strengthen the method for experimenter's medium vessels diastole of asymmetric diethylarginine (ADMA) rising, this method comprises this experimenter's administration of HMG-CoA reductase inhibitor and L-arginine.

19. the method for claim 17 or 18, wherein the HMG-CoA reductase inhibitor contains simvastatin.

20. the method for claim 17 or 18, wherein the L-arginine contains slow releasing preparation.

21. the method for claim 17 or 18, wherein this method improves endothelial function at least about 5 to about 15%.

22. the method for claim 17 or 18, wherein this method improves endothelial function at least about 7 to about 12%.

23. the method for claim 17 or 18, wherein this experimenter to suffer from endothelial function unusual.

24. increase the method that nitric oxide (NO) produces among the experimenter that asymmetric diethylarginine (ADMA) raises, this method comprises uses the L-arginine to this experimenter, wherein the L-arginine has overcome the inhibitory action of ADMA.

25. strengthen the method for experimenter's medium vessels diastole of asymmetric diethylarginine (ADMA) rising, this method comprises uses the L-arginine to this experimenter, wherein the L-arginine has overcome the inhibitory action of ADMA.

26. the method for claim 24 or 25, wherein this experimenter to suffer from endothelial function unusual.

27. the method for claim 24 or 25, wherein the L-arginine contains slow releasing preparation.

28. the method for claim 24 or 25, wherein this method makes endothelial function improve about 5 to about 15%.

29. the method for claim 24 or 25, wherein this method makes endothelial function improve about 6 to about 10%.

30. the method for claim 24 or 25, it also comprises the administration of HMG-CoA reductase inhibitor to the experimenter.

31. the method for claim 30, wherein the HMG-CoA reductase inhibitor contains simvastatin.

32. the method for claim 30, wherein administration of HMG-CoA reductase inhibitor and L-arginine make endothelial function improve about 5% to about 15%.

33. the method for claim 30, wherein administration of HMG-CoA reductase inhibitor and L-arginine make endothelial function improve about 7% to about 12%.

34. the sustained release l-arginine compositions, said composition contains:

(a) about by weight 25% to about 75%L-arginine or its pharmaceutically acceptable salt;

(b) about by weight 0.5% to about 5% polyvinylpyrrolidone;

(c) about by weight 5% to about 40% hydroxypropyl emthylcellulose;

(d) about by weight 2% to about 20% microcrystalline Cellulose;

(e) about by weight silicon dioxide below 3%; With

(f) about by weight magnesium stearate below 3%.

35. the compositions of claim 34, it contains

(a) about by weight 50%L-arginine mono-hydrochloric salts, wherein the L-arginine comprises L-arginine mono-hydrochloric salts;

(b) about by weight 3% to about 4% polyvinylpyrrolidone;

(c) about by weight 35% hydroxypropyl emthylcellulose;

(d) about by weight 10% microcrystalline Cellulose;

(e) about by weight colloidal silica below 1%, wherein silicon dioxide comprises colloidal silica; With

(f) about by weight magnesium stearate below 1%.

36. prepare the method for the arginic slow releasing composition of L-, this method comprises

(a) use granulating agent granulation L-arginine to form granule;

(b) moistening this granule of milling;

(c) dry this granule;

(d) drying this granule of milling; With

(e) this granule is mixed with at least a slow releasing agent.

37. the method for claim 36, wherein step (e) comprises the step of premixing, mixing and final mixing granules agent.

38. the method for claim 36, it is preceding with L-arginine and bonding agent dry mixed that it also is included in granulation step.

39. the method for claim 38, wherein bonding agent contains polyvinylpyrrolidone.

40. the method for claim 36, this method comprises

(a) with comprising the granulating agent granulation L-arginine of polyvinylpyrrolidone, wherein the L-arginine is the about 50% of slow releasing preparation by weight, and wherein polyvinylpyrrolidone is about 3% to about 4% of slow releasing preparation by weight;

(b) moistening this granule of milling;

(c) dry this granule;

(d) drying this granule of milling; With

(e) this granule is mixed with hydroxypropyl emthylcellulose, wherein hydroxypropyl emthylcellulose is about 35% of slow releasing preparation by weight.

41. the method for claim 40, this method also comprises mixes granule with microcrystalline Cellulose, colloidal silica and magnesium stearate, wherein microcrystalline Cellulose is about 10% of slow releasing preparation by weight, wherein colloidal silica is the about below 1% of slow releasing preparation by weight, and wherein magnesium stearate is the about below 1% of slow releasing preparation by weight.

42. be used for the treatment of or prevent the food stick of angiopathy or imbalance, this food stick contains and comprises the arginic slow releasing preparation of L-.

43. the food stick of claim 42, wherein slow releasing preparation contains the arginic slow release granule of L-.

44. the food stick of claim 42, it also contains the HMG-CoA reductase inhibitor.

45. the food stick of claim 42, wherein this food stick cholesterol reducing when being consumed by the experimenter.

46. the food stick of claim 42, wherein this food stick is used for the treatment of or prevents Alzheimer.

47. the food stick of claim 42, wherein this food stick is used for the treatment of or prevents intermittent claudication.

48. the food stick of claim 42, wherein this food stick reduces proteins C reactive when being consumed by the experimenter.

49. comprising the experimenter used, the method for prevention or treatment experimenter's medium vessels disease or imbalance, this method contain the food stick that comprises the arginic slow releasing preparation of L-.

50. reduce the method for cholesterol among the experimenter, this method comprises the experimenter used and contains the food stick that comprises the arginic slow releasing preparation of L-.

51. increase nitric oxide production method among the experimenter, this method comprises the experimenter used and contains the food stick that comprises the arginic slow releasing preparation of L-.

52. strengthen the method for experimenter's medium vessels diastole, this method comprises the experimenter used and contains the food stick that comprises the arginic slow releasing preparation of L-.

53. comprising the experimenter used, the method for Alzheimer among prevention or the treatment experimenter, this method contain the food stick that comprises the arginic slow releasing preparation of L-.

54. comprising the experimenter used, the method for prevention or the limping of treatment experimenter discontinuous, this method contain the food stick that comprises the arginic slow releasing preparation of L-.

55. reduce the method for proteins C reactive among the experimenter, this method comprises the experimenter used and contains the food stick that comprises the arginic slow releasing preparation of L-.

56. each method of claim 49 to 54, wherein food stick also contains the HMG-CoA reductase inhibitor.

57. the method for claim 56, wherein the HMG-CoA reductase inhibitor contains simvastatin.

58. reduce the method for cholesterol among the experimenter, this method comprises the experimenter used and contains the arginic slow releasing preparation of L-.

59. the method for claim 58, wherein this method reduces T-CHOL and low density lipoprotein, LDL (LDL) cholesterol among the experimenter.

60. the method for claim 58, wherein this method makes T-CHOL reduce about 50 to about 150mg/dL.

61. the method for claim 58, wherein this method makes T-CHOL reduce about 80 to about 100mg/dL.

62. the method for claim 58, wherein this method makes the LDL cholesterol reduce about 40 to about 110mg/dL.

63. the method for claim 58, wherein this method makes the LDL cholesterol reduce about 60 to about 100mg/dL.

64. the method for claim 58, wherein this method increases experimenter's middle-high density lipoprotein (HDL) cholesterol.

65. the method for claim 58, wherein this method makes triglyceride reduce about 30 to about 100mg/dL.

66. the method for claim 58, wherein this method makes triglyceride reduce about 45 to about 75mg/dL.

67. the method for claim 58, wherein slow releasing preparation contains

(a) about by weight 25% to about 75%L-arginine or its pharmaceutically acceptable salt;

(b) about by weight 0.5% to about 5% polyvinylpyrrolidone;

(c) about by weight 5% to about 40% hydroxypropyl emthylcellulose;

(d) about by weight 2% to about 20% microcrystalline Cellulose;

(e) about by weight silicon dioxide below 3%; With

(f) about by weight magnesium stearate below 3%.

68. the method for claim 67, wherein slow releasing preparation contains

(a) about by weight 50%L-arginine mono-hydrochloric salts, wherein the L-arginine comprises L-arginine mono-hydrochloric salts;

(b) about by weight 3% to about 4% polyvinylpyrrolidone;

(c) about by weight 35% hydroxypropyl emthylcellulose;

(d) about by weight 10% microcrystalline Cellulose;

(e) about by weight colloidal silica below 1%, wherein silicon dioxide comprises colloidal silica; With

(f) about by weight magnesium stearate below 1%.

69. the method for treatment or prevention angiopathy or imbalance, this method comprises uses slow releasing preparation to the experimenter, and this slow releasing preparation contains

(a) about by weight 25% to about 75%L-arginine or its pharmaceutically acceptable salt;

(b) about by weight 0.5% to about 5% polyvinylpyrrolidone;

(c) about by weight 5% to about 40% hydroxypropyl emthylcellulose;

(d) about by weight 2% to about 20% microcrystalline Cellulose;

(e) about by weight silicon dioxide below 3%; With

(f) about by weight magnesium stearate below 3%.

70. the method for claim 69, wherein slow releasing preparation contains

(a) about by weight 50%L-arginine mono-hydrochloric salts, wherein the L-arginine comprises L-arginine mono-hydrochloric salts;

(b) about by weight 3% to about 4% polyvinylpyrrolidone;

(c) about by weight 35% hydroxypropyl emthylcellulose;

(d) about by weight 10% microcrystalline Cellulose;

(e) about by weight colloidal silica below 1%, wherein silicon dioxide comprises colloidal silica; With

(f) about by weight magnesium stearate below 1%.

71. comprising the experimenter used, the method for treatment or prevention Alzheimer, this method contain the arginic slow releasing preparation of L-.

72. the method for claim 71, wherein slow releasing preparation contains

(a) about by weight 25% to about 75%L-arginine or its pharmaceutically acceptable salt;

(b) about by weight 0.5% to about 5% polyvinylpyrrolidone;

(c) about by weight 5% to about 40% hydroxypropyl emthylcellulose;

(d) about by weight 2% to about 20% microcrystalline Cellulose;

(e) about by weight below 3% silicon dioxide and

(f) about by weight magnesium stearate below 3%.

73. the method for claim 72, wherein slow releasing preparation contains

(a) about by weight 50%L-arginine mono-hydrochloric salts, wherein the L-arginine comprises L-arginine mono-hydrochloric salts;

(b) about by weight 3% to about 4% polyvinylpyrrolidone;

(c) about by weight 35% hydroxypropyl emthylcellulose;

(d) about by weight 10% microcrystalline Cellulose;

(e) about by weight colloidal silica below 1%, wherein silicon dioxide comprises colloidal silica; With

(f) about by weight magnesium stearate below 1%.

74. comprising the experimenter used, the method for treatment or prevention intermittent claudication, this method contain the arginic slow releasing preparation of L-.

75. the method for claim 74, wherein slow releasing preparation contains

(a) about by weight 25% to about 75%L-arginine or its pharmaceutically acceptable salt;

(b) about by weight 0.5% to about 5% polyvinylpyrrolidone;

(c) about by weight 5% to about 40% hydroxypropyl emthylcellulose;

(d) about by weight 2% to about 20% microcrystalline Cellulose;

(e) about by weight below 3% silicon dioxide and

(f) about by weight magnesium stearate below 3%.

76. the method for claim 75, wherein slow releasing preparation contains

(a) about by weight 50%L-arginine mono-hydrochloric salts, wherein the L-arginine comprises L-arginine mono-hydrochloric salts;

(b) about by weight 3% to about 4% polyvinylpyrrolidone;

(c) about by weight 35% hydroxypropyl emthylcellulose;

(d) about by weight 10% microcrystalline Cellulose;

(e) about by weight colloidal silica below 1%, wherein silicon dioxide comprises colloidal silica; With

(f) about by weight magnesium stearate below 1%.

77. treatment or the atherosis method of prevention experimenter's medium-sized artery, this method comprises uses slow releasing preparation to the experimenter, and this slow releasing preparation contains

(a) about by weight 25% to about 75%L-arginine or its pharmaceutically acceptable salt;

(b) about by weight 0.5% to about 5% polyvinylpyrrolidone;

(c) about by weight 5% to about 40% hydroxypropyl emthylcellulose;

(d) about by weight 2% to about 20% microcrystalline Cellulose;

(e) about by weight below 3% silicon dioxide and

(f) about by weight magnesium stearate below 3%.

78. the method for claim 77, wherein slow releasing preparation contains

(a) about by weight 50%L-arginine mono-hydrochloric salts, wherein the L-arginine comprises L-arginine mono-hydrochloric salts;

(b) about by weight 3% to about 4% polyvinylpyrrolidone;

(c) about by weight 35% hydroxypropyl emthylcellulose;

(d) about by weight 10% microcrystalline Cellulose;

(e) about by weight colloidal silica below 1%, wherein silicon dioxide comprises colloidal silica; With

(f) about by weight magnesium stearate below 1%.

79. strengthen the method for experimenter's medium vessels diastole, this method comprises uses slow releasing preparation to the experimenter, this slow releasing preparation contains

(a) about by weight 25% to about 75%L-arginine or its pharmaceutically acceptable salt;

(b) about by weight 0.5% to about 5% polyvinylpyrrolidone;

(c) about by weight 5% to about 40% hydroxypropyl emthylcellulose;

(d) about by weight 2% to about 20% microcrystalline Cellulose;

(e) about by weight below 3% silicon dioxide and

(f) about by weight magnesium stearate below 3%.

80. the method for claim 79, wherein slow releasing preparation contains

(a) about by weight 50%L-arginine mono-hydrochloric salts, wherein the L-arginine comprises L-arginine mono-hydrochloric salts;

(b) about by weight 3% to about 4% polyvinylpyrrolidone;

(c) about by weight 35% hydroxypropyl emthylcellulose;

(d) about by weight 10% microcrystalline Cellulose;

(e) about by weight colloidal silica below 1%, wherein silicon dioxide comprises colloidal silica; With

(f) about by weight magnesium stearate below 1%.

81. increase among the experimenter-method that nitrogen oxide produces, this method comprises uses slow releasing preparation to the experimenter, this slow releasing preparation contains

(a) about by weight 25% to about 75%L-arginine or its pharmaceutically acceptable salt;

(b) about by weight 0.5% to about 5% polyvinylpyrrolidone;

(c) about by weight 5% to about 40% hydroxypropyl emthylcellulose;

(d) about by weight 2% to about 20% microcrystalline Cellulose;

(e) about by weight below 3% silicon dioxide and

(f) about by weight magnesium stearate below 3%.

82. the method for claim 81, wherein slow releasing preparation contains

(a) about by weight 50%L-arginine mono-hydrochloric salts, wherein the L-arginine comprises L-arginine mono-hydrochloric salts;

(b) about by weight 3% to about 4% polyvinylpyrrolidone;

(c) about by weight 35% hydroxypropyl emthylcellulose;

(d) about by weight 10% microcrystalline Cellulose;

(e) about by weight colloidal silica below 1%, wherein silicon dioxide comprises colloidal silica; With

(f) about by weight magnesium stearate below 1%.

83. reduce the method for proteins C reactive, this method comprises uses the L-arginine to the experimenter.

84. the method for claim 83, wherein the L-arginine comprises slow releasing preparation.

85. the method for claim 83, wherein slow releasing preparation contains

(a) about by weight 25% to about 75%L-arginine or its pharmaceutically acceptable salt;

(b) about by weight 0.5% to about 5% polyvinylpyrrolidone;

(c) about by weight 5% to about 40% hydroxypropyl emthylcellulose;

(d) about by weight 2% to about 20% microcrystalline Cellulose;

(e) about by weight below 3% silicon dioxide and

(f) about by weight magnesium stearate below 3%.

86. the method for claim 85, wherein slow releasing preparation contains

(a) about by weight 50%L-arginine mono-hydrochloric salts, wherein the L-arginine comprises L-arginine mono-hydrochloric salts;

(b) about by weight 3% to about 4% polyvinylpyrrolidone;

(c) about by weight 35% hydroxypropyl emthylcellulose;

(d) about by weight 10% microcrystalline Cellulose;

(e) about by weight colloidal silica below 1%, wherein silicon dioxide comprises colloidal silica; With

(f) about by weight magnesium stearate below 1%.

87. reduce the method for proteins C reactive among the experimenter, this method comprises experimenter's administration of HMG-CoA reductase inhibitor and contains the arginic slow releasing preparation of L-.

88. the method for claim 87, wherein this method makes proteins C reactive reduce about 10% to about 50%.

89. the method for claim 87, wherein this method makes proteins C reactive reduce about 25% to about 35%.

90. the method for claim 87, wherein this method with do not make when experimenter's administration of HMG-CoA reductase inhibitor not being used compare when containing the arginic slow releasing preparation of L-that proteins C reactive reduces about 50% to about 90%.

91. the method for claim 87, wherein this method with do not make when experimenter's administration of HMG-CoA reductase inhibitor not being used compare when containing the arginic slow releasing preparation of L-that proteins C reactive reduces about 65% to about 75%.

92. the method for claim 87, wherein this method with do not make when experimenter's administration of HMG-CoA reductase inhibitor not being used compare when containing the arginic slow releasing preparation of L-that proteins C reactive reduces about 80% to about 120%.

93. the method for claim 87, wherein this method with do not make when experimenter's administration of HMG-CoA reductase inhibitor not being used compare when containing the arginic slow releasing preparation of L-that proteins C reactive reduces about 95% to about 105%.

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