CN104739844A - Ursodesoxycholic acid and use thereof - Google Patents
Ursodesoxycholic acid and use thereof Download PDFInfo
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- CN104739844A CN104739844A CN201310731260.1A CN201310731260A CN104739844A CN 104739844 A CN104739844 A CN 104739844A CN 201310731260 A CN201310731260 A CN 201310731260A CN 104739844 A CN104739844 A CN 104739844A
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Abstract
The invention relates to chenodeoxycholic acid and a use thereof. The invention concretely comprises uses of the compound shown in the formula (I) in preparation of drugs for resisting platelet aggregation and thrombogenesis and preventing and treating platelet abnormal activation-related diseases. The chenodeoxycholic acid has effects of resisting platelet aggregation and spreading, can be used as a novel platelet aggregation-resistant and antithrombotic drug for treating blood platelet abnormal activation-related diseases such as thrombotic diseases, hypercoagulable state viral diseases, tumors, coronary heart diseases, apoplexy, hypertension, abdominal aortic aneurysm, leukemia and disseminated intravascular coagulation (DIC).
Description
Technical field
The invention belongs to technical field of pharmaceuticals, be specifically related to ursodesoxycholic acid [Ursodeoxycholic Acid (the UDCA)] novelty teabag in antiplatelet aggregation and the application in prevention and therapy arterial thrombotic disease thereof.
Technical background
Along with growth in the living standard, the arterial thrombotic disease such as apoplexy, coronary heart disease has become the number one killer threatening China and developed country's people ' s health.Therefore, very important to the control of cardiovascular and cerebrovascular vessel thrombotic disease.Blood platelet disorders activate the pathologic basis that the Intravascular Thrombus formation caused is arterial thrombotic disease coronary heart disease and apoplexy, its process comprises hematoblastic gathering and sprawls, therefore suppresses the antiplatelet drug of platelet activation to be prevention and the Main Means for the treatment of this kind of disease.The antiplatelet drug of extensive use clinically includes cox-2 inhibitors aspirin, thiophene pyridines P2Y at present
12receptor antagonist clopidogrel (clopidogrel), prasugrel (prasugrel), platelet fibrinogen receptor antagonist tirofiban hydrochloride (TirofibanHydrochloride) and phosphodiesterase inhibitor cilostazol (Cilostazol).These medicines or there is gastrointestinal toxicity, or there is the side effect such as bleeding tendency, and in clinical practice, have tolerance and leptin suppression, limit to their application, made the therapeutic effect of coronary heart disease and apoplexy still not completely as good as one wishes.Platelet has specific morphosis and biochemical composition, has more constant quantity in normal blood.They with inactive form at body internal recycle, until they touch the defective region of endotheliocyte or run into coagulation cascade reaction and just activated.Now they attach to the defect of endotheliocyte, deformation occur, release particles content, and mutually adhere to formation aggregation.
When blood vessel is damaged, platelet sustain damage position activate factor stimulate occur hematoblastic gathering, become platelet plug, play primary hemostatic effect, then platelet produces thrombin through complicated change again, make the Fibrinogen in contiguous blood plasma become fibrin, the fibrin interweaved mutually makes platelet plug tangle into blood clot with hemocyte, i.e. thrombosis.Simultaneously hematoblastic projection stretches in fibrin net, and along with the contraction of platelet microfilament (actin) and myosin, blood clot is shunk, and thrombosis becomes more solid, more effectively can play anastalsis, and this is the anastalsis of secondary.Along with the formation of thrombosis, platelet can discharge a lot of cell active substance, comprise dense granule, α granule and platelet derived microparticle, wherein containing various active materials such as ADP, 5-hydroxy tryptamine, platelet factor 4, beta-thromboglobulin, thrombospondin, cell growth factor, blooc coagulation factor V, VII, Ⅻ and vascular permeability factors, these active substances are by activating Peripheral blood platelet, promote vasoconstriction, the various ways such as short fibrin formation strengthen haemostatic effect.Some material then can strengthen inflammation and the immunoreation of damage location.In pathological conditions, when blood platelet disorders activate generation gathering, the thrombotic disease such as apoplexy, coronary heart disease will be caused.
Due to the tremulous pulse supplying brain blood, apoplexy mainly occurs that atherosis and platelet aggregation forms thrombosis, make luminal stenosis even inaccessible, cause focal cerebral.Fall ill because of acute brain blood supply insufficiency.And coronary heart disease is to break or rotten to the corn due to IC atherosclerotic plaque, activate platelet adhesion, gathering and being formed caused by thrombosis.Therefore, anticoagulant forms the core link that thrombosis has become prevention and therapy apoplexy and coronary heart disease, and the target spot of intervention should be platelet, thrombin, established fibrin and other blood factors.In addition hematoblastic abnormal aggregation take part in the course of disease of the disease such as viral disease, tumor disease, coronary heart disease, apoplexy, hypertension, leukemia, disseminated inravascular coagulation (DIC) with hypercoagulability.Antiplatelet (i.e. anticoagulant) medicine has positive impact to these diseases.
Ursodesoxycholic acid (3 α, 7 beta-dihydroxy-5 β-cholanic acids, UDCA) is a kind of chenodiol, in the total bile of the mankind, account for 3%.Be separated from Chinese Fel Ursi by the Shoda of Japanese Okayama Univ. the earliest and name, Leuschner in 1985 etc. find that serum transaminase reduces when merging the cholelithiasis of hepatitis with its treatment, and after this large amount of clinical research also demonstrate that UDCA has definite therapeutical effect to some hepatopathys.UDCA is a kind of avirulent hydrophilic cholic acid, and toxic endogenous cholic acid can be suppressed competitively in the absorption of ileum.By activating the signal network of calcium ion, Protein kinase C composition, and strengthening the hepatocellular secretion capacity of cholestasis by activating mitotic activity protein kinase, the hydrophobic Bile acid concentrations of endogenous in blood and hepatocyte being reduced, reaches the effect of anti-cholestasis.UDCA can also replace the toxicity cholic acid molecules on cell membrane and organelle competitively, prevents hepatocyte and bile duct cell to be subject to the infringement of more toxicity cholic acid.Above-mentioned effect is in particular in: (1) cytoprotection.UDCA obviously can alleviate the hepatocellular cytolysis that hydrophobic cholic acid brings out, the apoptosis that minimizing cultivation Mus and human hepatocytes are brought out by toxicity cholic acid.(2) membrane stabilizing action.Gland Mitochondria Membrane permeability that UDCA can prevent cholic acid from bringing out changes, that is by gland Mitochondria Membrane, basement membrane and little bile duct membrane damage that membrane stabilizing action prevents toxicity cholic acid from bringing out.(3) antioxidation.The Kupffer Cell that UDCA can suppress toxicity cholic acid to cause activates, and can also increase hepatocyte glutathion and the level containing mercaptan albumen, prevent hepatocellular oxidative damage.(4) immunoregulation effect.UDCA is suppressed indirectly by the stimulation reducing hydrophobic cholic acid, and directly suppresses the expression of histocompatibility complex (MHC) I class and II genoid by activating glucocorticoid receptor (GR).
Ursodesoxycholic acid can increase bile acid secretion, and bile component is changed, reduce bile cholesterol and cholesterol ester, the cholesterol be conducive in cholelithiasis dissolves gradually, for the cholesterol calculus of unsuitable operative treatment, but bile pigment calculus, combination calculus and radiopaque calculus can not be dissolved.To cholecystitis, biliary tract inflammation and dyspepsia also have certain curative effect.The purposes of ursodesoxycholic acid in cardiovascular is not yet reported.Late Cambrian ursodesoxycholic acid of the present invention has the new role that antiplatelet aggregative activity and antiplatelet stretch in vitro simultaneously, and oral after there is obvious antiplatelet effects.
Summary of the invention
The invention provides the new opplication of ursodesoxycholic acid in antiplatelet aggregation and thrombosis, its advantage is: experiment in vitro display can suppress the platelet aggregation of multiple agonist induction, and there is the effect that antiplatelet sprawls, Mouse oral ursodesoxycholic acid experiment display its there is good platelet effects.
Specifically, the chemistry of ursodesoxycholic acid of the present invention (Chenodeoxycholic acid (CDCA)) is called 3a, and 7 beta-dihydroxy-5 β-cholestane-24-acid, it has the structure of formula I, and its chemical formula is: molecular formula C
24h
40o
4, molecular weight 392.58.
Based on above discovery, the invention provides the new opplication of formula I compound ursodesoxycholic acid in antiplatelet aggregation and thrombosis, its advantage is the platelet aggregation that experiment in vitro display can suppress multiple agonist induction, and there is the effect that antiplatelet sprawls, Mouse oral ursodesoxycholic acid experiment display has good platelet effects.
In one embodiment, the invention provides the pharmaceutical composition comprising formula I compound.In one embodiment, described pharmaceutical composition also comprises any activity or inactive composition.In one embodiment, described pharmaceutical composition also comprises pharmaceutically acceptable excipient.
In one embodiment, pharmaceutical composition ursodesoxycholic acid of the present invention is 5 μMs-100 μMs.In one embodiment, pharmaceutical composition ursodesoxycholic acid of the present invention is 100 μMs.
In one embodiment, the invention provides the purposes of compound in antiplatelet aggregation of formula I.In one embodiment, the invention provides the purposes of compound in the medicine preparing antiplatelet aggregation of formula I.In one embodiment, medicine provided by the invention can suppress hematoblastic gathering and sprawl.
In another embodiment, the invention provides the purposes of compound in antithrombus formation of formula I.In another embodiment, the compound of formula I of the present invention is used for the treatment of thrombosis.In another embodiment, the invention provides the purposes of compound in the antithrombotic medicine of preparation of formula I.
In another embodiment, the compound that the invention provides formula I to activate the purposes in relevant disease with blood platelet disorders in prevention or treatment.In another embodiment, the compound of formula I of the present invention activates relevant disease for preventing or treating with blood platelet disorders.In another embodiment, the compound that the invention provides formula I to activate the purposes in the medicine of relevant disease with blood platelet disorders in preparation prevention or treatment.In one embodiment, of the present inventionly activate that relevant disease includes but not limited to thrombotic disease, has the viral disease of hypercoagulability, tumor disease, coronary heart disease, apoplexy, hypertension, abdominal aortic aneurysm, leukemia, disseminated inravascular coagulation (DIC) with blood platelet disorders.
In one embodiment, medicine of the present invention or compound be prepared into applicable oral, intravenous, intraperitoneal, locally, transdermal, through eye, per nasal, locally, suck, subcutaneous, intramuscular, suck, the dosage form of Sublingual or rectally.In one embodiment, pharmaceutical dosage form of the present invention includes but not limited to tablet, capsule, pill, granular preparation, injection.
In one embodiment, medicine of the present invention or compound can be prepared or administering drug combinations with any activity or inactive ingredient combination.In one embodiment, active component of the present invention comprises aspirin, chlorine than Gray or cilostazol.
Accompanying drawing explanation
Fig. 1 is the inhibition (congregational rate figure) to the platelet aggregation of ADP induction after ursodesoxycholic acid oral administration.Which show the platelet of matched group and ursodesoxycholic acid oral administration group mice, the hematoblastic gathering for ADP10 μM of induction has different reactivities, and the aggregation rate of ursodesoxycholic acid oral administration group mouse platelets is starkly lower than matched group.
Detailed description of the invention
Before description materials and methods of the present invention, should be appreciated that and the invention is not restricted to described concrete grammar, scheme, material and reagent, because they can be different.Will also be understood that term used herein only for describing the object of detailed description of the invention, and and the scope of the invention that only limited by the non-provisional application submitted to subsequently of not intended to be limiting.
Unless otherwise defined, otherwise all technology used herein and scientific terminology have the identical meaning usually understood with those skilled in the art.Implement although can adopt to those similar or equivalent any methods described herein and material or test the present invention, what describe at present is preferred method and material.The all publications specifically mentioned herein and patent are included in by reference herein for all objects, comprise report in description and public publication can with the chemical substance of coupling of the present invention, instrument, statistical analysis and method.The all lists of references quoted in this description should regard as the technical merit showing this area.All the elements all should not be construed as and admit instinct of the present invention by means of formerly invention prior to these disclosures herein.
I. summarize
The implication that as used herein, term " comprises ", " comprising " and its equivalents comprise " containing " and " by ... composition ", the compositions such as " comprising " X can only be made up of X maybe can contain other material, such as X+Y.
Clearly indicate unless separately had in context, otherwise herein and the singulative " " used in claims, " one " and " being somebody's turn to do " comprise plural referents.Equally, term " one " (or " one "), " one or more " and " at least one " are used interchangeably herein.
The present invention relates generally to a kind of medicine " ursodesoxycholic acid " with formula I structure in the novelty teabag of antiplatelet aggregation and application thereof, provide the platelet aggregation inhibitory activity experiment of ursodesoxycholic acid, be included in the multiple agonist (ADP of vitro inhibition, thrombin, collagen) platelet aggregation of inducing, suppress hematoblastic sprawling, the gathering of oral rear suppression mouse platelets.Its effect that there is antiplatelet aggregation and sprawl of Late Cambrian of the present invention, the present invention is also contained ursodesoxycholic acid and is used for the treatment of blood platelet disorders as new antiplatelet aggregation and antithrombotic reagent and activates relevant disease, includes but not limited to thrombotic disease, has the viral disease of hypercoagulability, tumor disease, coronary heart disease, apoplexy, hypertension, abdominal aortic aneurysm, leukemia, disseminated inravascular coagulation (DIC) etc.
As used herein, term " separation " refers to that material is separated from its primal environment (if crude, namely primal environment is natural surroundings).As the polynucleotide under the native state in living cells and albumen be do not have separative, but same polynucleotide or albumen as from native state with in other materials existed separately, then for be separated.
As used herein, term " platelet " refers to it is one of visible component in mammalian.Out-of-shape, acellular core, generally rounded, volume is less than erythrocyte and leukocyte, and in adult's blood, platelet counts is 1 × 10
11~ 3 × 10
11individual/L.Platelet has specific morphosis and biochemical composition, more constant quantity is had in normal blood, there is the physiological functions such as adhesion, gathering, release, play an important role in the physiology such as hemostasis, wound healing, inflammatory reaction, thrombosis and organ-graft refection and pathological process.
As used herein, " thrombosis " refers in cardiovascular, the process that blood solidifies or in blood, some visible component is separated out, coagulation forms solid matter block.The solid matter block formed is called thrombosis.Thrombosis is formed under the state of blood flow.
As used herein, term " disease " and " disease " are used interchangeably, and they are all reflected infringement normal function, usually shows by S&S and cause the abnormality of one of the human or animal body or its part of life-span of human or animal or quality of life reduction.Disease described herein refers to comprise the disease relevant with platelet aggregation, as thrombotic disease, the viral disease with hypercoagulability, tumor disease, coronary heart disease, apoplexy, hypertension, abdominal aortic aneurysm, and leukemia, disseminated inravascular coagulation (DIC) etc.
As used herein, term " gathering " refers to platelet adhesion each other, usually point two phases.First assembles phase is also reversible aggregation phase; Second assembles phase is also irreversible aggrengation phase.
As used herein, term " agonist " has guided the material of platelet aggregation, also cries and causes poly-agent or derivant.Agonist of the present invention comprises pathologic and causes poly-agent such as virus, antibacterial, immune complex, medicine etc. and cause poly-agent as ADP(adenosine diphosphate (ADP) with physiological), TXA2. (Thromboxane A2), collagen, thrombin.
ADP of the present invention is that most important physiological causes poly-agent, especially the endogenous ADP that goes out of intra platelet free calcium.The ADP of low concentration only causes reversible aggregation phase, and platelet is assembled rapidly and depolymerization rapidly.Add the ADP of median dose in platelet suspension after, platelet is assembled rapidly, then depolymerization; In initiation reversible aggregation mutually soon after, platelet is assembled again, after this no longer depolymerization, and this irreversible aggrengation it is believed that it is caused by intra platelet free calcium endogenous ADP mutually.The ADP of high concentration directly causes irreversible aggrengation phase.If by platelet suspension in the liquid a few hours not containing glucose, or add the medicine suppressing ATP metabolism, or add calcium chelating agent, then can suppress the aggreation that ADP causes.ADP cannot induce clean platelet (eliminating Fibrinogen) to occur assembling.The aggreation that ADP causes has dose dependent, and needs calcium ion, fibrinogenic participation.
After collagen of the present invention and platelets contact, after experiencing a lag phase, directly enter irreversible aggrengation phase.This may be because collagen is induced aggregation reacts while, also triggers intra platelet free calcium ADP, TXA2. etc.
Thrombin of the present invention has the dose dependent similar with ADP; Aggreation unlike thrombin induction does not need fibrinogenic participation.
As used herein, term " is sprawled " and is referred to platelet adhesion at vascular injury site, cause final metamorphosis (becoming flat from spheroidal) by the change of cytoskeletal protein, increase contact area, be acknowledged as key one step of Thrombosis and hemostasis.
As used herein, term " suppression " refers to the control action that medicine or compounds on platelet are assembled, sprawled." suppression ratio " described herein and " gathering suppression ratio " is used interchangeably, and they are the processed group percentage ratio that platelet aggregation degree reduces compared with solvent control group.
As used herein, term " effective dose " or " treatment effective dose " show give object with treat imbalance, disease or disease time the amount of compound be enough to carry out this type for the treatment of for condition of illness or disease or disease." treatment effective dose " changes according to following factor: the judgement of compound, the imbalance for the treatment of or disease or morbid state, the imbalance for the treatment of or the order of severity of disease or disease, the age of object and relative health, route of administration and form, nursing medical worker or veterinary and other factors.In the present invention, " effective dose " or " treatment effective dose " can refer to that about 0.01 milligram of per kilogram of body weight every day (mg/kg/ day) is to about 500mg/kg/ day, preferably about 0.1 milligram of per kilogram of body weight every day (mg/kg/ day) is to about 300mg/kg/ day, more preferably about 1 milligram of per kilogram of body weight every day (mg/kg/ day) arrives about 100mg/kg/ day.Preferred amount can be determined by those skilled in the art.Such as, the doctor in charge easily can define effective amount by using conventional method and observing the result obtained in a similar situation.When determining the effective dose of compound of the present invention, many factors are considered by the doctor in charge, include but not limited to particular compound to be administered; If the co-administered of other medicament used; Mammiferous kind; Its size, age and general health; The severity of the tight disease of disease; The reaction of individual patient; Administering mode; The bioavailability characteristics of the preparation of institute's administration; Selected dosage; The use of other concomitant drugs; And other relevant situation.
As used herein, term " carrier " or " supporting agent " describe the composition in pharmaceutical composition or preparation except active pharmaceutical compounds.Carrier of the present invention includes but not limited to lactose, microcrystalline Cellulose, starch, magnesium stearate, PVP, carboxymethyl starch sodium, polyvinylpolypyrrolidone, hyprolose, silicon dioxide and ethanol.
The present invention relates to and give patient by compound to treat effective dose.Compound can give separately or a part as pharmaceutically acceptable compositions gives.In addition, compound or compositions such as disposablely all can being given by injecting, such as repeatedly to be given by a series of tablet or such as by using transdermal delivery substantially to send equably within a period of time.In addition, the dosage of compound can time to time change.Instant release type preparation, controlled release formulation or its combination can be used to give compound.
Pharmaceutical composition of the present invention can single unit dose or with multiple single unit dose preparation, packaging or large quantities of sale.As used herein, " unit dose " is for comprising the discrete amount of the pharmaceutical composition of the active component of scheduled volume.The amount of active component is generally equal to the dosage of the active component by giving patient or the adequate rate of this dosage, such as, and 1/2nd of this dosage or 1/3rd.
In one embodiment, compound of the present invention is individually dosed.In another embodiment, the compounds of this invention and other active or inactive ingredient combination administration, in another embodiment, the compounds of this invention is prepared with other activity or inactive ingredient combination.Active component of the present invention includes but not limited to oxidase inhibitor (as aspirin), thiophene pyridines P2Y12 receptor antagonist (as clopidogrel), platelet fibrinogen receptor antagonist (example hydrochloric acid tirofiban) and phosphodiesterase inhibitor (as cilostazol).
Kind according to treated people, stature and disease change by the relative quantity of the active component in pharmaceutical composition of the present invention, pharmaceutically acceptable carrier and any supplementary element, and change according to described the giving approach of compositions.For example, described compositions can comprise the active component of 0.1%-100% (w/w).Or pharmaceutical composition of the present invention comprises 10 μMs of-300 μMs of ursodesoxycholic acid, preferably 10 μMs, 30 μMs, 50 μMs, 75 μMs, 150 μMs, 300 μMs ursodesoxycholic acid.In one embodiment, pharmaceutical composition of the present invention comprises 100 μMs of ursodesoxycholic acid.
As used herein, " giving " and " administration " comprises compound introducing health of the present invention, preferably introduces any mode of systemic circulation.Compound of the present invention can carry out administration by any effective way, include but not limited to oral, intravenous, intraperitoneal, locally, transdermal, through eye, per nasal, locally, suck, subcutaneous, intramuscular, suck, Sublingual, rectally etc.
Pharmaceutical composition of the present invention can be prepared into any suitable dosage form, such as tablet, capsule, pill, granular preparation, injection, suppository, aerosol, effervescent tablet, drop, drop pill, eye drop, nasal drop, ophthalmic ointment, gargarism, sublingual tablet, sticking tablet, membranous patch, solution, lotion, liniment, ointment, plaster, paste etc.Dosage form of the present invention also comprises solid or injectable implant or storage agent.
The compositions being suitable for injecting comprises the active component with the upper acceptable aseptic aqueous solution of pharmaceutically acceptable carrier such as physiology or non-aqueous solution, dispersion, suspension or Emulsion coupling, maybe can comprise the sterilized powder for redissolving in sterile injectable solution or dispersion.The example of suitable aqueous and non-aqueous carrier, diluent, solvent or supporting agent comprises water, isotonic saline solution, ethanol, polyhydric alcohol (propylene glycol, Polyethylene Glycol, glycerol etc.), its suitable mixture, triglyceride, comprise vegetable oil as olive oil, or injectable organic ester is as ethyl oleate.Can such as by use coating as lecithin if dispersion then by keep desired particle size and/or by use surfactant keep suitable mobility.This type of preparation can be suitable for injecting the form given or give continuously and prepare, packs or sell.Injectable formulation can unit dosage forms preparation, packaging or sell, such as ampoule bottle, comprise antiseptic multi-dose container or for automatic injection or the single operative installations injected by doctor.
Pharmaceutical composition is also prepared with the form of injectable sterile aqueous or oiliness (Emulsion) suspension or solution, pack or is sold.This suspension or solution can be prepared according to technology known in the art, and also can comprise supplementary element, such as dispersant as herein described, wetting agent or suspending agent except active component.Such as, the acceptable non-toxic diluent of parenteral or solvent such as water or 1,3 butylene glycol can be used to prepare this type of sterile injectable preparation.Other acceptable diluent and solvent comprise Ringer's solution, isotonic sodium chlorrde solution and non-volatile oils, such as, synthesize monoglyceride or two glyceride.The acceptable preparation of other useful parenteral comprises those preparations containing active component, and described active component is microcrystalline form, in Liposomal formulation or as the part of biodegradable polymers system.Compositions for slow release or implantation can comprise pharmaceutically acceptable polymerization or hydrophobic material, such as Emulsion, ion exchange resin, slightly soluble polymer or slightly soluble salt.
Also can comprise adjuvant according to compound of the present invention, such as antiseptic, wetting agent, emulsifying agent and/or dispersant, comprise such as P-hydroxybenzoic acid fat, chlorobutanol, phenol, sorbic acid etc.Also may need to comprise isotonic agent, such as sugar, sodium chloride etc.By the Long-term absorption using the medicament such as aluminum monostearate and/or gelatin that can postpone to absorb to cause Injectable composition.Specifically, wherein can use liposome, micelle and emulsifying agent, thus make the compounds of this invention be easier to dissolve to send.
Capsule, tablet, powder and granule is comprised for the oral solid dosage forms given.In this type of solid dosage forms, reactive compound mixes with at least one conventional inert excipients (or carrier), described excipients such as, as sodium citrate or dicalcium phosphate or (a) filler or extender, starch, lactose, sucrose, mannitol or silicic acid; (b) binding agent, such as carboxymethyl cellulose, alginate, gelatin, polyvinylpyrrolidone, sucrose or Radix Acaciae senegalis; (c) wetting agent, such as glycerol; (d) disintegrating agent, such as, agar, calcium carbonate, Rhizoma Solani tuber osi or tapioca, alginic acid, some composition silicate or sodium carbonate; (e) solution retarder, such as paraffin; F () absorbs accelerator, such as quaternary ammonium compound; (g) wetting agent, such as spermol or glyceryl monostearate; (h) adsorbent, such as Kaolin or bentonite; And/or (i) lubricant, such as Talcum, calcium stearate, magnesium stearate, solid polyethylene glycol, sodium lauryl sulfate or their mixture.When capsule and tablet, dosage form also can comprise buffer agent.
Tablet can such as assign to prepare by compression or moulding activated one-tenth, and it optionally comprises one or more supplementary elements.By active component boil down to free-flowing form (as powder or granular preparation) being prepared compressed tablets in suitable device, described active component optionally mixes with one or more in binding agent, lubricant, excipient, surfactant and dispersant.Molded tablet is prepared by being molded active component, pharmaceutically acceptable carrier and the mixture of liquid that is at least enough to wet mix in suitable device.
Pharmaceutically acceptable excipient for the production of tablet comprises inert diluent, granulating agent and disintegrating agent, binding agent and lubricant.Known dispersant comprises potato starch and sodium starch glycolate.Known surfactant comprises sodium lauryl sulfate.Known diluent comprises calcium carbonate, sodium carbonate, lactose, microcrystalline Cellulose, calcium phosphate, calcium hydrogen phosphate and sodium phosphate.Known granulating agent and disintegrating agent comprise corn starch and alginic acid.Known binding agent comprises gelatin, Radix Acaciae senegalis, pregelatinised corn starch, polyvinylpyrrolidone and hydroxypropyl emthylcellulose.Known lubricant comprises magnesium stearate, stearic acid, Silicon stone and Talcum.
Tablet can not coated or its known method coating can be used to realize delay disintegrate in human gastrointestinal tract, thus make active component sustained release and absorption.For example, the material of such as glyceryl monostearate or glycerol distearate can be used for carrying out coating to tablet.Tablet also can comprise sweeting agent, flavoring agent, stain, antiseptic or these certain combination, to provide pharmaceutically high-quality with good to eat preparation.
Can prepare and there is coating or shell, such as the solid dosage forms of enteric coating and other coating well known in the art, as tablet, dragee, capsule and granule.It also can comprise opacifier, and can have and make it with the compositions of delayed mode release of active compounds.The example of available embedding composition is polymer and wax.Reactive compound can be also micro-encapsulated form, has one or more in above-mentioned excipient time suitable.
The solid composite of similar type can use as lactose or toffee and high molecular weight polyethylene glycol etc. as the capsule of the soft of excipient or hard gelatine filling in be used as filler.Can use and compositions such as the gelatin of physiologic degradation can prepare the hard capsules comprising active component.This type of hard capsules comprises active component, and can comprise supplementary element, comprises such as inert solid diluent as calcium carbonate, calcium phosphate or Kaolin.Can use and compositions such as the gelatin of physiologic degradation can prepare the soft gelatin capsule comprising active component.This type of soft capsule comprises can with water or oily medium as the active component of Oleum Arachidis hypogaeae semen, liquid paraffin or mixed with olive oil.
Known technology can be used to prepare oral cavity composition, and it takes at the small intestinal of human patients or large enteral liberation port the medicament given specially.Such as, preparation for delivery to gastronintestinal system (comprising colon) comprises enteric coating system, its based on such as only at the methacrylate copolymer of pH6 and above dissolving as poly-(methacrylic acid, methyl methacrylate), thus described polymer only just starts when entering small intestinal to dissolve.The amount of small intestine transit speed and contained polymer is depended in the position of described polymer formulations disintegrate.Such as, relatively thick polymer coating is for delivery to colon near-end.Also can use the polymer that can provide location specific colonic delivery, wherein said polymer depends on large intestine bacterium colony to provide the enzymatic degradation of polymer coating thus to discharge medicine.Such as, the polysaccharide of azobenzene polymer, glucoside and various natural acquisition and modification can be used in this type of preparation.
Pharmaceutically acceptable Emulsion, solution, suspension, syrup and elixir is comprised for the oral liquid dosage form given.Except reactive compound, liquid dosage form can comprise inert diluent such as water or other solvent conventional in this area, isotonic saline solution, solubilizing agent and emulsifying agent, such as ethanol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzylalcohol, benzyl benzoate, propylene glycol, 1, 3-butanediol, dimethyl formamide, oils, be specially almond oil, Oleum Arachidis hypogaeae semen, Oleum Cocois, Oleum Gossypii semen, Oleum Arachidis hypogaeae semen, maize embryo oil, olive oil, Oleum Ricini, Oleum sesami, glycerol, the vegetable oil of fractional distillation, mineral oil is as liquid paraffin, tetrahydrofurfuryl alcohol, Polyethylene Glycol, the mixture etc. of the fatty acid ester of sorbitan or these materials.
Except these inert diluents, compound of the present invention also can comprise adjuvant as wetting agent, emulsifying agent and suspending agent, demulcent, antiseptic, buffer agent, salt, sweeting agent, flavoring agent, stain and aromatic.Except reactive compound, suspension also can comprise suspending agents if the edible fat of ethoxylated isostearyl alcohols, polyoxyethylene sorbitol ester or sorbitan ester, microcrystalline Cellulose, hydrogenation, sodium alginate, polyvinylpyrrolidone, Tragacanth, Radix Acaciae senegalis, agar and cellulose derivative are as the mixture etc. of sodium carboxymethyl cellulose, methylcellulose, hydroxypropyl emthylcellulose, partially aluminium hydroxide, bentonite or these materials.Be suitable for the oral pharmaceutical composition of the present invention given liquid preparation can in liquid form or the form of the dry products redissolved with water or another suitable supporting agent before use with purport prepare, pack and sell.
Known dispersant or wetting agent comprise naturally occurring phospholipid as lecithin, alkylene oxide and fatty acid, with long chain aliphatic, with derived from fatty acid and the partial ester of hexitol or the condensation product (being such as respectively Myrj 45, heptadecyl ethyleneoxy spermol, polyoxyethylene 80 sorbitan monooleate and Polysorbate 80) with the partial ester derived from fatty acid and hexitan.Known emulsifying agent comprises lecithin and Radix Acaciae senegalis.Known antiseptic comprises methyl, ethyl or n-pro-pyl-p-hydroxy Benzoic Acid, ascorbic acid and sorbic acid.Known sweeting agent comprises such as, glycerol, propylene glycol, sorbitol, sucrose and glucide.The known thickening agent for oily suspensions comprises such as Cera Flava, hard paraffin and spermol.
The liquid solution of active component in aqueous or oil-based solvent can be substantially identical with liquid suspension mode be prepared, main difference be active component be dissolve but not suspend in a solvent.The liquid solution of pharmaceutical composition of the present invention can comprise about each component described in liquid suspension, should understand suspending agent and may not contribute to active component dissolving in a solvent.Aqueous solvent comprises such as water and isotonic saline solution.Oil-based solvent comprises such as almond oil, oily ester, ethanol, vegetable oil if the vegetable oil of Oleum Arachidis hypogaeae semen, olive oil, Oleum sesami or Oleum Cocois, fractional distillation and mineral oil are as liquid paraffin.
Compositions of the present invention also can comprise cyclodextrin composition such as to improve the water solublity of active pharmaceutical ingredient, prolong drug discharges and improves tabletting characteristics.Usually, the starch digestion thing of circulus oligomer (" cyclodextrin ") some antibacterial of glucose.The abundantest cyclodextrin is α, β and γ cyclodextrin respectively with 6,7 and 8 glucose units.The inner chamber of cyclodextrin is hydrophobic and the exposure of molecule is hydrophilic.Known cyclodextrin can improve active pharmaceutical ingredient stability, water solublity reduce volatility.Some examples of commercially available cyclodextrin or derivatives thereof are as follows: alpha-cyclodextrin; (2-hydroxypropyl)-alpha-cyclodextrin; Beta-schardinger dextrin-; 6-O-α-D-glucose group-beta-cyclodextrin; Gamma-cyclodextrin; (2-hydroxypropyl)-gamma-cyclodextrin.
The compositions given for rectum or vagina is by preparing compound of the present invention and any additional compound as Oleum Cocois, Polyethylene Glycol or suppository wax mix with suitable non-irritating excipient or carrier, it is liquid under body temperature in solid at normal temperatures, thus dissolves and release of active ingredients in rectum or vaginal cavity.Such composition can be such as suppository, be detained the form of enema agent and rectum or colon irrigation solutions.Suppository formulations also can comprise various supplementary element, comprises antioxidant and antiseptic.Be detained enema agent or rectum or colon irrigation solutions by active component and pharmaceutically acceptable liquid-carrier being combined to prepare.As known in the art, enema agent can utilize and can be packaged in be suitable for people's rectum structure delivery apparatus in use.Enema agent also can comprise various supplementary element, comprises antioxidant and antiseptic.
Pharmaceutical composition of the present invention can be suitable for the preparation that vagina gives and prepare, packs or sell.Such composition can be that the material of such as the inserted vagina of suppository, dipping or coating is as tampon, the form of rinsing preparation or Vaginal lavages solution.
The dosage form giving the compounds of this invention for local comprises unguentum, powder, spraying and inhalant.Compound aseptically acceptable carrier upper with physiology mixes, and may need any antiseptic, buffer and/or propellant.Be suitable for the preparation that gives of local and comprise liquid or semi-liquid preparations if liniment, lotion, oil-in-water or water in oil emulsion are as ointment, unguentum or paste, and solution or suspension.The preparation that gives of local also can comprise one or more of supplementary element described herein.
Ophthalmic preparations, eye ointment, powder and solution are also considered to contain within the scope of the present invention.This type of preparation can such as in the form of eye drop, the solution of the active component including effective amount in aqueous or oil-based liquid carrier or suspension.This type of drop also can comprise in buffer agent, salt or supplementary element described herein one or more.In other embodiments, the preparation that eye can give comprises in microcrystalline form or in Liposomal formulation active component.
The pharmaceutical composition of the present invention that preparation is used for pulmonary delivery the form of solution or suspension drop can provide active component.This type of preparation can be used as to be prepared containing the optionally aseptic aqueous of active component or dilute alcohol solution or suspension, pack or sells, and any atomization or sprayer unit can be utilized to give easily.This type of preparation also can comprise one or more supplementary elements, comprises flavoring agent if saccharin sodium, volatile oil, buffer agent, surfactant or antiseptic are as methyl hydroxybenzoate.The drop provided by this route of administration preferably has the average diameter in about 0.1-about 200 nanometer.
Pharmaceutical composition of the present invention can be suitable for the preparation that buccal gives and prepare, packs or sell.This type of formulation example such as using conventional procedures makes the form of tablet or lozenge, and such as can include the active component of effective amount, containing mouth-soluble solution or degradation composition and the remainder optionally in one or more supplementary elements described herein.Or, be suitable for preparation that buccal gives and can comprise containing the powder of active component or the solution of aerosolization or atomization or suspension.This type of is Powdered, aerosolization or atomization preparation preferably have when disperseing mean diameter or the droplet size that about 0.1-is about 200nm, and can comprise supplementary element described herein one or more.
In order to use in non-human animal, compound of the present invention can paste or pellet form preparation and give as usual implant below animal head or ear skin.Paste formulations is by being dispersed in pharmaceutically acceptable oil as prepared in Oleum Arachidis hypogaeae semen, Oleum sesami, Semen Maydis wet goods by compound.The pellet comprising the compound for the treatment of effective dose mixes prepare by compound and diluent is clung to wax etc. as carbowax, Carlow, and can add lubricant if magnesium stearate or calcium stearate are to improve prilling.Certainly, recognize and can give animal to reach required dosage level by more than a kind of pellet.In addition, found also can during treatment of animals by this type of implant periodically to being kept active levels suitable in animal body.
The dosage range of compound of the present invention and pharmaceutically acceptable salt thereof can change as required, and this depends on age of institute's treatment target and quality, expection route of administration, the compound etc. that specifically gives.Benefit from dosage range and optimal dose that those of ordinary skill in the art of the present disclosure have the ability to determine concrete patient.It is also noted that compound of the present invention can be used in the preparation of slow release, controlled release and delayed release, these dosage forms are also known by those of ordinary skill in the art.In one embodiment, pharmaceutical composition of the present invention comprises 10 μMs of-300 μMs of ursodesoxycholic acid.In one embodiment, pharmaceutical composition of the present invention comprises 300 μMs of ursodesoxycholic acid.
Whether the compounds of this invention directly gives cell, celliferous tissue, the body fluid of exposing cell or compound therefrom spreads or the position that is transported to cell is not critical.Give patient with certain amount and approach by compound thus the amount of described compound to be enough to make the lipid in cell directly or indirectly arrive at cell by this approach just enough.Minimum changes with the feature of compound.
Spendable concrete dosage and dosage range depend on many factors, comprise the demand of patient, the sanatory order of severity and give the pharmacologically active of compound.According to the disclosure, to the dosage range of concrete patient and the determination of optimal dose completely within the ordinary skill of those skilled in the art.The compound prescription that gengral practitioner, dentist or veterinary determine being easy to and be with effective amount should be understood, to move lipid storage in patients, to cause losing weight or appetite-suppressing.In the preamble, first doctor or veterinary such as can output comparatively low dosage prescription, increase dosage subsequently until obtain suitably response.But will also be understood that, concrete dosage level for any unique individual will depend on many factors, comprise the activity of particular compound used, age of people, body weight, general health situation, sex and diet, administration time, route of administration, secreting rate, any drug regimen and treat the order of severity of imbalance.
When preparing with medicine injectable dosage formulations (comprising the combination of described herein and claimed compound and injectable carrier system), compound of the present invention is particularly useful.As used herein, injectable and infusion dosage form (i.e. parenteral dosage form) include but not limited to lipidosome injection or the double-layer of lipoid vesicle of the phospholipid contained for encapsulating active medicine.Injection is included in the sterile product that parenteral uses.
There are five kinds of different types of injections of USP definition: Emulsion, lipid, powder, solution and suspension.Emulsion injection agent comprises the aseptic Emulsion without thermal source preparation containing being intended to parenteral and giving.Be intended to redissolve for the lipid complex of injection of solution agent and powder the sterile preparation forming the solution that parenteral uses.Powder for suspension injection is intended to redissolve the sterile preparation forming the suspension that parenteral uses.Freeze-dried powder for liposome suspension injection is intended to redissolve the sterile cryo drying agent used with parenteral, its manner of formulation allows to include liposome in as containing the double-layer of lipoid vesicle for active drug substance being encapsulated in the phospholipid in double-layer of lipoid or aqueous space, with this, preparation can be formed after redissolution.Freeze-dried powder for injection of solution agent is the solution prepared by lyophilizing (" lyophilization ") dosage form used, described process comprises the moisture removed with pole low-pressure in freezing state product thus, and all meets with the generation that adds of this liquid subsequently the solution that injection requires in all respects.Freeze-dried powder for suspension injection is intended to the liquid preparation that parenteral uses, and it comprises the solid be suspended in suitable liquid medium, and it all meets the requirement of sterile suspensions in all respects, carrys out supending medicament used with this by lyophilizing.Injection of solution agent comprises liquid preparation, its comprise one or more be dissolved in be suitable for inject suitable solvent or mutually miscible solvent mixture in medicine.Solution concentrate injection comprise parenteral use sterile preparation, its produce after adding suitable solvent all in all meet injection require solution.Suspension injection comprises (being suitable for injecting) liquid preparation, it comprises the solid particle be scattered in whole liquid phase, and described granule is insoluble, and with this, oil phase is scattered in whole aqueous phase, vice versa.Suspension lipidosome injection is (being suitable for injection) liquid preparation, and its mode formed with liposome (usually comprising the double-layer of lipoid vesicle of the phospholipid for encapsulating active drug material in double-layer of lipoid or aqueous space) is scattered in the oil phase of whole aqueous phase.The ultrasonic injection of suspension is (the being suitable for injecting) liquid preparation comprising the solid particle be scattered in whole liquid phase, and described granule is insoluble.In addition, owing to gas sparging to be produced the microsphere formed by solid particle by suspension, therefore supersound process can be carried out to product.
The present composition also can comprise one or more pharmaceutically suitable excipient, as solvent and cosolvent, solubilizing agent, wetting agent, suspending agent, thickening agent, emulsifying agent, chelating agen, buffer, pH adjusting agent, antioxidant, reducing agent, anti-microbial preservative, extender, protective agent, tension regulator and special additive.
II. ursodesoxycholic acid is to the inhibition of the human platelet aggregation that ADP induces
In one embodiment, ursodesoxycholic acid has inhibition to the human platelet aggregation that ADP induces.Ursodesoxycholic acid concentration dependent suppresses the human platelet aggregation of ADP10 μM of induction, ursodesoxycholic acid 10 μMs of suppression ratio are 9 ± 3%, ursodesoxycholic acid 30 μMs of suppression ratio are 21 ± 5%, ursodesoxycholic acid 50 μMs of suppression ratio are 34 ± 4%, ursodesoxycholic acid 75 μMs of suppression ratio are 46 ± 3%, ursodesoxycholic acid 150 μMs of suppression ratio are 69 ± 5%, and ursodesoxycholic acid 300 μMs of suppression ratio are 95 ± 8%.(see table 1).
III. ursodesoxycholic acid is to the inhibition of the human platelet aggregation of other multiple agonist inductions
In another embodiment, ursodesoxycholic acid 150 μMs has inhibition to the human platelet aggregation of other multiple agonist inductions: when 150 μMs, ursodesoxycholic acid can Trombin inhibiting and collagen-induced platelet aggregation, the suppression ratio of platelet aggregation of inducing thrombin 0.1U/ml is 17% ± 6%, is 19% ± 4% to the suppression ratio of the platelet aggregation that collagen 1 μ g/ml induces.(see table 2)
IV. ursodesoxycholic acid inhibition that human blood platelets is sprawled
In another embodiment, ursodesoxycholic acid is sprawled human blood platelets has inhibition: ursodesoxycholic acid concentration dependent suppresses human blood platelets to be sprawled, 75 μMs of ursodesoxycholic acid suppression ratio are 26 ± 4%, 150 μMs of ursodesoxycholic acid suppression ratio are 35 ± 3%, 300 μMs of ursodesoxycholic acid suppression ratio is 48 ± 6%.(see table 3)
V. the antiplatelet aggregation effect after oral ursodesoxycholic acid
In another embodiment, oral give ursodesoxycholic acid after there is antiplatelet aggregation effect: in ursodesoxycholic acid oral (300mg/kg/day) administration one week mice, platelet aggregation rate is measured after abdominal aortic blood, ursodesoxycholic acid administration significantly suppresses hematoblastic gathering in one week, and it is 21% ± 7% to the platelet aggregation inhibition rate of ADP10 μM of induction.(see Fig. 1, table 4)
Experimental result confirms, compound ursodesoxycholic acid shown in formula I of the present invention has good inhibition for the human platelet aggregation of multiple agonist induction, to platelet sprawling on slide, there is good inhibition, finally after oral administration in mice, find that ursodesoxycholic acid has remarkable platelet effects (namely anticoagulant forms thrombosis).On this basis, inventor finds ursodesoxycholic acid, and a kind of traditional treatment liver and gall diseases medicine, at the novelty teabag of prevention and therapy cardiovascular disease.It is by anticoagulant, suppresses platelet to be sprawled, and plays good platelet effects (namely anticoagulant forms thrombosis).To the thrombotic cardiovascular disease that pathomechanism is blood platelet disorders activation, there is prevention and therapy effect.
Embodiment
Below in conjunction with specific embodiment, set forth the present invention further.Should be understood that these embodiments are only not used in for illustration of the present invention to limit the scope of the invention by any way.The experimental technique of unreceipted actual conditions in the following example, usual conveniently condition is as people such as Alan D.Michelson, in the condition described in " platelet " second edition (NewYork:Elsevier Press, 2007), or according to the condition that manufacturer advises.Unless otherwise indicated, in literary composition use reagent from traditional Chinese medicines reagent company limited, percentage ratio and number are calculated by weight.By above-mentioned explanation and following examples, illustrate herein and describe those outside of the present invention various amendment it will be apparent to those skilled in the art that and fall within the scope of the appended claims.
Unless otherwise defined, all specialties and scientific words and one skilled in the art the same meaning be familiar with.In addition, any method similar or impartial to described content and material all can be applicable in the present invention.The use that better implementation method described in literary composition and material only present a demonstration.
Embodiment 1. ursodesoxycholic acid is to the inhibition of the human platelet aggregation that ADP induces
The object of the present embodiment is the human platelet aggregation proving that ursodesoxycholic acid can suppress ADP to induce.
In the present embodiment, to be hematoblasticly prepared as follows: platelet comes from the healthy volunteer signing Informed Consent Form, volunteer is getting non-Aspirin in first 20 days of blood, and chlorine is than any antiplatelet drugs such as Grays.Anticoagulant adopts ACD (85mmol L
-1sodium citrate, 71.38mmol L
-1citric acid, and 27.78mmolL
-1glucose).300x g obtains platelet rich plasma in centrifugal 20 minutes, and after getting upper strata platelet rich plasma, 900x g obtains platelet in centrifugal 10 minutes further, finally uses resuspended (the 138mmol L of Tyrode ' s buffer
-1sodium chloride (NaCl), 2.7mmol L
-1potassium chloride (KCl), 2mmol L
-1magnesium chloride (MgCl
2), 0.42mmol L
-1sodium dihydrogen phosphate (NaH
2pO
4), 5mmol L
-1glucose (glucose), 10mmol L
-1hEPES, 0.2% bovine serum albumin (BSA), and 0.02unit mL
-1apyrase (apyrase), pH7.4, obtain washed platelet, number of platelets is adjusted to 2.5 × 10
8individual platelet/ml.
Hematoblastic gathering measures: be hematoblasticly gathered in platelet aggregation instrument (Model400VS, Chrono-Log company, Haverston, PA), parameter setting: mixing speed 900rpm, temperature 37 DEG C, before adding agonist ADP, platelet solvent or medical preconditioning 3 minutes, curve of platelet aggregation monitoring time is 5 minutes, and assembling suppression ratio is the processed group percentage ratio that platelet aggregation degree reduces compared with solvent control group.
Assemble suppression and the results are shown in Table 1, ursodesoxycholic acid (Sigma, sigma, USA), the human platelet aggregation that concentration dependent suppresses ADP10 μM (Chrono-log company, Havertown, PA) to induce.
Table 1: ursodesoxycholic acid concentration dependent suppresses the human platelet aggregation of ADP5 μM of induction.
| Ursodesoxycholic acid dosage (μM) | Platelet aggregation inhibition rate (n=3) |
| 10 | 9±3％ |
| 30 | 21±5％ |
| 50 | 34±4％ |
| 75 | 46±3％ |
| 150 | 69±5％ |
| 300 | 95±8％ |
Embodiment 2. ursodesoxycholic acid 150 μMs of inhibitions to the human platelet aggregation of other multiple agonist inductions
The object of the present embodiment proves that ursodesoxycholic acid can Trombin inhibiting and collagen-induced human platelet aggregation.
In the present embodiment, hematoblastic preparation and hematoblastic aggregation inducing and assay method are with embodiment 1.The results are shown in Table 2, ursodesoxycholic acid is Trombin inhibiting (Chrono-log company, Havertown, PA) (0.1U/mL) 150 μMs time, and the human platelet aggregation that collagen (Chrono-log company, Havertown, PA) (1 μ g/mL) is induced.
Table 2: ursodesoxycholic acid Trombin inhibiting and collagen-induced human platelet aggregation.
| Ursodeoxycholic acid concentration (μM) | Agonist title (concentration) | Platelet aggregation inhibition rate (n=3) |
| 150 | Thrombin (0.1U/mL) | 17％±6％ |
| 150 | Collagen (1 μ g/mL) | 19％±4％ |
The inhibition that embodiment 3. ursodesoxycholic acid is sprawled for platelet.
The object of the present embodiment proves that ursodesoxycholic acid can suppress platelet to be sprawled.
In the present embodiment, hematoblastic preparation is with embodiment 1, and it is as follows that platelet sprawls experimentation:
1) the bag quilt of coverslip or culture plate: by coverslip or batch cultur plate 20 μ g/mL Fibrinogen (fibrinogen, Chrono-log company, Havertown, PA) (be dissolved in PBS) and wrap quilt, 4 DEG C are spent the night, 2% bovine serum albumin 37 DEG C closed nonspecific binding site 1h, PBS rinse once.
2) be 1.8 × 10 by the platelet HEPES-Tyrode buffer of matched group or ursodesoxycholic acid processed group adjustment concentration
6/ 500 μ L, be inoculated on the coverslip of 24 orifice plate inner fibrin primordial coverings, every hole 500 μ L, hatches 2 hours for 37 DEG C.
3) above-mentioned coverslip is taken out, wash once with pre-cooling PBS, stick the fixing 20min of platelet 4% paraformaldehyde (formaldehyde).
4) the PBS room temperature of the platelet fixed containing 3%BSA is closed 30min.
5) PBS containing 1%BSA and microfilament green fluorescence probe (actin-Tracker Green, green skies company, China) is added, incubated at room 45min.
6) PBS develops a film 5 times, each 3min, deionized water rinsing coverslip 1 time, to remove unnecessary salinity.
7) dry coverslip, microscope slide drips the anti-cancellation mountant of fluorescence (DAKO company), coverslip is inverted and is covered in mountant, avoid occurring bubble.Use nial polish mounting, 4 DEG C keep in Dark Place.
8) fluorescence microscope (Leica DM5500, Japan) observed result.The each high power field platelet of meter record sprawls situation.
Above step all notably lucifuge.The results are shown in Table 3, ursodesoxycholic acid concentration dependent suppresses sprawling of human blood platelets.
Table 3: ursodesoxycholic acid concentration dependent suppresses sprawling of human blood platelets.
| Ursodeoxycholic acid concentration (μM) | Platelet sprawls suppression ratio (n=3) |
| 75μM | 26±4％ |
| 150μM | 35±3％ |
| 300μM | 48±6％ |
Anticoagulant effect after the oral administration of embodiment 4. ursodesoxycholic acid
The object of the present embodiment is can anticoagulant after proving ursodesoxycholic acid oral administration.
In the present embodiment, 6 to 8 ages in week mice (BALB/C) oral ursodesoxycholic acid (300mg/kg/day is dissolved in ethanol) and contrast (ethanol) administration after a week, 10% chloral hydrate (400mg kg
-1) anesthesia, external acquisition mouse platelets, with 2.9% sodium citrate (sodium citrate: whole blood=1:9) anticoagulant, 300x g obtains platelet rich plasma in centrifugal 5 minutes, after getting upper strata platelet rich plasma, 900x g obtains platelet in centrifugal 8 minutes further, final Tyrode ' s buffer is resuspended, and obtain washed platelet, number of platelets is adjusted to 2.5 × 10
8individual platelet/ml.Hematoblastic gathering assay method is with embodiment 1.The results are shown in Figure 1 and table 4.Significantly can suppress ADP(10 μM after the administration of ursodesoxycholic acid Mouse oral) platelet aggregation of inducing, show its oral after still there is good anti-platelet activity.
Table 4: the oral rear suppression mouse platelets of ursodesoxycholic acid is assembled.
Each pertinent literature listed above all introduces the application as a reference with a full section.Many aspects involved in the present invention have been done and have as above been set forth.It is to be understood, however, that, under the prerequisite of the spirit and scope that do not depart from the present invention, be all allow to any modification of foregoing description.Equally, similar situation is also included within claim.
Claims (10)
1. the purposes of the compound of formula I in the medicine preparing antiplatelet aggregation
2. purposes as claimed in claim 1, is characterized in that, the hematoblastic gathering of described Drug inhibition and sprawling.
3. the purposes of compound in the antithrombotic medicine of preparation of formula I
4. the compound of formula I is preparing the purposes preventing to activate with blood platelet disorders in the medicine of relevant disease
5. the compound of formula I to activate the purposes in the medicine of relevant disease with blood platelet disorders in preparation treatment
6. the purposes as described in claim 4 or 5, it is characterized in that, describedly activate that relevant disease comprises thrombotic disease, has the viral disease of hypercoagulability with blood platelet disorders, tumor disease, coronary heart disease, apoplexy, hypertension, abdominal aortic aneurysm, leukemia or disseminated inravascular coagulation (DIC).
7. the purposes according to any one of claim 1-5, is characterized in that, described medicine be prepared into applicable oral, intravenous, intraperitoneal, locally, transdermal, through eye, per nasal, suction, subcutaneous, intramuscular, suck, the dosage form of Sublingual or rectally.
8. purposes as claimed in claim 7, it is characterized in that, described dosage form comprises tablet, capsule, pill, granular preparation or injection.
9. the purposes according to any one of claim 1-5 is characterized in that, described medicine is prepared or administering drug combinations with any activity or inactive ingredient combination.
10. purposes as claimed in claim 9, is characterized in that described active component comprises aspirin, chlorine than Gray or cilostazol.
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Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2000026300A (en) * | 1998-07-02 | 2000-01-25 | Pola Chem Ind Inc | Pharmaceutical composition for protecting vascular endothelial cell |
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2013
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2000026300A (en) * | 1998-07-02 | 2000-01-25 | Pola Chem Ind Inc | Pharmaceutical composition for protecting vascular endothelial cell |
Non-Patent Citations (2)
| Title |
|---|
| 丁涛等: "精制熊胆粉活血化瘀作用研究", 《中国天然药物》 * |
| 陈瑞兰等: "熊胆粉化学成分及药理作用研究进展", 《辽宁中医药大学学报》 * |
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