CN105968113B - A kind of triazolopyrimidine derivative and its application - Google Patents

Abstract

The present invention provides novel triazolopyrimidines derivative and its crystal form and preparation method shown in a kind of formula II and they preparing anticoagulation, antithrombotic, treat or prevent cerebral ischemia diseases or improve sleep drug in application.

Description

A kind of triazolopyrimidine derivative and its application

Technical field

The present invention relates to organic chemistry and pharmaceutical field, and in particular to a kind of triazolopyrimidine derivative and its crystal form and system Preparation Method, and preparing anticoagulation, antithrombotic, treat or prevent cerebral ischemia diseases or improving answering in the drug slept With.

Background technique

Thrombus is to be formed in coagulation process by platelet aggregation, and the thrombus formed in non-damaging situation can drop Low blood flow velocity blocks tail vein even so as to cause cerebral arterial thrombosis, tissue necrosis, athero- artery sclerosis, cardiac muscle stalk It waits indefinitely disease.

Cerebral arterial thrombosis refer to the brain tissue part feeding artery blood perfusion that occurs suddenly reduce or blood flow it is complete in It is disconnected, stop blood supply, oxygen supply, for sugar etc., destroys local brain tissue disintegration.The main reason for cerebral arterial thrombosis are as follows: 1. move Thromboembolism caused by pulse atherosclerosis；2. cerebral embolism caused by cardiac-derived embolus；3. vasculitis caused by a variety of causes, blood Pipe damage and wound etc..Cerebral arterial thrombosis is generally fallen ill in nighttime sleep, and Chang Weici morning finds limb adynamia when getting up Or hemiplegia, how unconscious obstacle, blood pressure can be normal or higher, can there is artery sclerosis history.Cerebral arterial thrombosis accounts for cerebral apoplexy patient The 60%~70% of sum mainly includes cerebral thrombosis and cerebral embolism.The cohesion of blood platelet is oftenCerebral thrombosisWhat is formed opens End, therefore the drug of platelet aggregation is prevented to can be used to treat or prevent cerebral arterial thrombosis.

There are many approach and mechanism for the activation of blood platelet, and wherein P2Y12 receptor participates in fibrinogen deceptor activation, blood The processes such as the platelet aggregation that bolt is formed, thromboxane A2 generates, wound causes.The P2Y12 receptor of the mankind is by 342 amino acid Composition, is distributed mainly in blood platelet and brain tissue.When the endogenous stimulus factor such as ADP etc. activates P2Y12 receptor, can swash The accesses such as PI3K living, and then Rap1b, Akt, ERK access are activated, cause the activation of fibrinogen deceptor jointly, to activate Fibrinogen causes thrombosis or platelet aggregation.This process must just may be used in the case where P2Y12 receptor activation To realize, therefore block P2Y12 receptor that can significantly inhibit by the platelet aggregation and blood of ADP and the initiation of other stimulating factors Bolt is formed.

Currently, the whole world, which has multiple P2Y12 acceptor inhibitors, successfully develops listing, wherein by AstraZeneca (AstraZeneca) ticagrelor (Ticagrelor) of exploitation listing is the first reversible P2Y12 inhibitor in the whole world, 2010 December obtains listing approval in European Union for the first time, and 2 months 2011 in Britain's Initial Public Offering；In August, 2011 is also approved to list in the U.S. Approval, in the country's listing of more than 40, the whole world.Ticagrelor is clinically mainly used to reduce acute coronary syndrome (ACS) incidence of the thrombotic cardiovascular event of patient.The usage and dosage of ticagrelor are as follows: oral, initial dose 180mg, Maintenance dose is adjusted to twice a day, each 90mg；Treatment time generally at least 12 months.

Ticagrelor chemical name are as follows: (1S, 2S, 3R, 5S) -3- [7- { [(1R, 2S) -2- (3,4- difluorophenyl) cyclopropyl] Amino } -5- rosickyite base -3H-1,2,3- triazol [4,5-d] pyrimidin-3-yl] -5- (2- hydroxyl-oxethyl) -1,2- ring penta 2 Alcohol, structure can be prepared as shown in formula I by method disclosed in CN1432017A.

Ticagrelor has the unique chemical moieties of triazolo pyrimidine, can be directly in conjunction with P2Y12 receptor invertibity, not only It can quickly play and inhibit platelet aggregation effect, and in 24 hours of final dose, with the decline of blood concentration, suppression Production rapid decay, platelet function also fast quick-recovery therewith.Therefore, ticagrelor is that one kind can quick acting, potent suppression The new oral antiplatelet drug of platelet aggregation processed, good security, undoubtedly clinically acute coronary syndrome (ACS) the more preferable of patient's Antiplatelet therapy is selected.

But ticagrelor is there are still some shortcomings, such as lower (the ticagrelor biology benefit in human body of bioavilability Expenditure average out to 36%), medication compliance not high (in long-term administration, needing medication in one day twice, increase misses possibility) etc.. Therefore, it is necessary to develop the new derivative of ticagrelor, to keep ticagrelor as reversible P2Y12 inhibitor advantage On the basis of, further increase bioavilability, enhancing medication compliance.

Physical and chemical properties of drugs or internal pharmacokinetic property can be improved by reasonable base group modification, improve oral life Object availability, this is known in those skilled in the art.Although theoretically can be reasonable according to the chemical functional group in molecule The modification of design assumption, but what female medicine generated after base group modification is completely new molecular entity, and which often shows The harmful physical chemistry or bioactive properties that female medicine is not present.Although base group modification compound seems " simple ", due to people The complexity of body and drug interaction, identification have physicochemical property appropriate, pharmacokinetic property, in vivo conversion and safety Property be complicated multidisciplinary task, therefore the base group modification compound obtained in practice often through " rational design ", often with Expected result differs greatly, or even obtains physicochemical property or the worse base group modification compound of mother's biological activity ratio medicine.Therefore, Being developed by base group modification improves the noval chemical compound of physical and chemical properties of drugs or internal pharmacokinetic property have very much can not Predictability, the solid state characterizations such as crystal form of these noval chemical compounds, new application are even more unpredictable.

The present invention surprisingly has found a kind of through base of ticagrelor in the research process to ticagrelor base group modification The novel triazolopyrimidines derivative of group's modification, the compound activity is suitable with ticagrelor, and oral absorption is better than ticagrelor, It is metabolizable at ticagrelor in vivo, to extend action time, provided clinically to reduce administration number of times, improving compliance Good material base；Furthermore the compound can exist with the solid-state form of crystal form, be conducive to production, storage and preparation Preparation；In addition, the compound not only has the function of anticoagulation, antithrombotic but also can be used to treat or prevent cerebral ischemia Property disease or improve sleep.

Summary of the invention

It is an object of the present invention to provide a kind of novel triazolopyrimidines derivative, which not only has anticoagulant The effect of blood, antithrombotic can be used to treat or prevent cerebral ischemia diseases or improve sleep, and pharmacokinetic property is better than Ticagrelor.

Another object of the present invention is to provide the preparation methods of the novel triazolopyrimidines derivative.

Another object of the present invention is to provide the system of a kind of crystal form of the novel triazolopyrimidines derivative He the crystal form Preparation Method.

Another object of the present invention is to provide the pharmaceutical composition comprising the novel triazolopyrimidines derivative or its crystal form Object.

Another object of the present invention is to provide the novel triazolopyrimidines derivative or its crystal form prepare anticoagulation or Purposes in the drug of antithrombotic.

Another object of the present invention is to provide the novel triazolopyrimidines derivative or its crystal form in preparation treatment or pre- Purposes in anti-cerebral ischemia diseases or the drug of improvement sleep.

In order to achieve the above-mentioned object of the invention, present invention firstly provides triazolopyrimidine derivative shown in a kind of formula II or Its officinal salt, eutectic, hydrate or solvate,

The definition of " salt " be it is well known to those skilled in the art of the present technique, refer to by cation and anion pass through ion The effect of key and the compound formed.

" eutectic " (Co-Crystals) refers to a kind of multicomponent crystal with fixed stoichiometric ratio, in the crystalline substance Each component is to be coexisted with molecular level by the effect combination of hydrogen bond or other non-covalent bonds, nonionic key in body.In drug In eutectic, active pharmaceutical ingredient and one or more eutectic forming bodies (Co-crystal former) are generally comprised.When independent Pure eutectic forming body at room temperature with liquid in the presence of, the eutectic be also referred to as " solvate ", wherein solvent be water when quilt Referred to as " hydrate "." eutectic " further includes the multicomponent crystal more in this way with fixed stoichiometric ratio, in these crystalline substances Between body pharmaceutical active ingredient and other components, a part is passed through by hydrogen bond or other non-covalent bond effects, another part Ionic bond or active force between hydrogen bond and ionic bond and combine.

" salt " or " eutectic " further include the forms such as the solvate of salt or eutectic, hydrate.When salt or eutectic are at certain In kind solvent when preparation, pulp or crystallization, which is likely to enter in salt or eutectic crystal, forms solvate；When this When solvent is water, that is, it is likely to form hydrate.

Determine " eutectic " or " salt " method be it is well known to those skilled in the art of the present technique, such as with Advances in crystal X-ray diffraction divide Analysis etc..

Purpose according to the present invention, the present invention provides triazolopyrimidine derivative shown in formula II or its officinal salt, The preparation method of eutectic, hydrate or solvate, this method comprises:

(1), III compound of formula is reacted into V compound of production with IV compound of formula or its sour addition product,

In formula III, R₁For F, Cl, Br, I, OH or OA, wherein A is hydroxy activated base；

(2), V compound of formula is deprotected to obtain II compound of formula,

(3), optional, salt is made in II compound of formula as needed, eutectic is made, hydrate is made or solvent is made and closes Object.

In above method step (1), " hydroxy activated base " in III compound of formula refers to such a group, introduces After the group, reactivity of the hydroxyl as leaving group, including sulfonyl, sulfinyl, hydrocarbon acyl group etc. can be improved, specifically Such as mesyl, trifyl, p-toluenesulfonyl, trifluoroacetyl group.

In above method step (1), " the sour addition product " refer to a certain acid compound by ionic bond, hydrogen bond or its The substance, including salt, eutectic, solvate etc. that the effect of his non-covalent bond is combined with another compound.IV chemical combination of formula Object is the common intermediate for preparing ticagrelor, commercially viable to buy or be made by literature method.IV idic acid of formula adds It include inorganic acid or organic acid at the acid in object；Wherein suitable inorganic acid is selected from hydrochloric acid, hydrobromic acid, hydroiodic acid, nitric acid, phosphoric acid Or sulfuric acid etc.；Suitable organic acid is selected from L-TARTARIC ACID, dibenzoyl-L-tartaric, two pairs of toluyl groups-L- winestones Acid, R-MA, R- α-methoxyphenylacetic acid, fumaric acid, D-malic acid, D- camphorsulfonic acid, S- ketone group pinic acid, oxalic acid, acetic acid, Trifluoroacetic acid, methanesulfonic acid, benzene sulfonic acid, p-methyl benzenesulfonic acid etc..

In above method step (1), the acid of a certain amount of alkali neutralization reaction generation need to generally be added or by IV compound of formula The acid that sour addition product introduces.The alkali includes organic base or inorganic base；Wherein suitable organic base include tertiary amine (such as triethylamine, Diisopropyl ethyl amine etc.)；Suitable inorganic base includes hydroxide (such as lithium hydroxide, hydroxide of alkali metal, alkaline-earth metal Sodium, potassium hydroxide etc.), carbonate (such as lithium carbonate, sodium carbonate, potassium carbonate), bicarbonate (such as sodium bicarbonate, saleratus Deng), phosphate (such as sodium phosphate, potassium phosphate), hydrophosphate (such as dibastic sodium phosphate, potassium hydrogen phosphate) etc..

In above method step (1), as R in III compound of formula₁When for OH, which can further comprise first by OH Activation is OA (A is hydroxy activated base, is defined as above), then the process reacted with IV compound of formula or its sour addition product.

In above method step (1), reaction dissolvent is chosen as being immiscible in the mixed solvent of the solvent of water and water, wherein institute It states and is immiscible in the solvent of water and can be selected from toluene, methylene chloride etc. or their mixture；Reaction dissolvent is also chosen as non-proton Solvent, wherein the aprotic solvent be selected from can acetonitrile, toluene, methylene chloride, acetone, dimethoxy-ethane, tetrahydrofuran, two Six ring of oxygen, N-Methyl pyrrolidone, N,N-dimethylformamide, DMAC N,N' dimethyl acetamide, hexamethyl phosphoramide, dimethyl are sub- Sulfone, sulfolane etc. or their mixture.

In above method step (1), the molar ratio of III compound of formula and IV compound of formula or its sour addition product Generally 0.7:1 to 1.3:1.

In above method step (1), reaction temperature is generally -10 DEG C to solvent boiling point.

In above method step (2), the reagent of the deprotection is generally acid, is selected from organic acid and inorganic acid；Suitable Organic acid is selected from methanesulfonic acid or trifluoroacetic acid etc.；Suitable inorganic acid is selected from hydrochloric acid, hydrobromic acid, hydroiodic acid or sulfuric acid etc..

In above method step (2), reaction dissolvent generally comprises the mixed solvent of water and organic solvent composition, wherein organic Solvent is selected from methanol, ethyl alcohol, isopropanol, the tert-butyl alcohol, tetrahydrofuran, dioxane, acetonitrile, acetone, toluene, methylene chloride etc..

In above method step (2), reaction temperature is generally 0 DEG C to solvent boiling point.

In above method step (1) or (2), the method that this field routine can be used in the determination in reaction time is carried out, and is such as adopted With TLC, HPLC monitoring etc..

In above method step (1), the preparation method preparation that III compound of formula can provide through the invention, this method Include:

(a), it reacts VI compound of formula or its sour addition product to obtain VIII compound of formula with VII compound of formula,

R in formula VII and VIII₁It is defined as above, the R in formula VII₂For F, Cl, Br, I or-OA, A is defined as above；

(b), VIII compound of formula is converted into III compound of an accepted way of doing sth in the presence of diazo reagent,

In formula III and VIII, R₁It is defined as above.

In above method step (a), the acid in the sour addition product of VI compound of formula includes inorganic acid or organic acid；Its In be suitable for inorganic acid be selected from hydrochloric acid, hydrobromic acid, hydroiodic acid, nitric acid, phosphoric acid or sulfuric acid etc.；Suitable organic acid is selected from L- winestone Acid, dibenzoyl-L-tartaric, two pairs of toluyl groups-L-TARTARIC ACIDs, oxalic acid, acetic acid, trifluoroacetic acid, methanesulfonic acid, benzene sulphur Acid, p-methyl benzenesulfonic acid etc..

In above method step (a), the molar ratio of VII compound of formula and VI compound of formula or its sour addition product Generally 3:1 to 0.8:1.

In above method step (a), the acid of a certain amount of alkali neutralization reaction generation need to generally be added or by VI compound of formula The acid that sour addition product introduces.The alkali includes organic base or inorganic base；Wherein suitable organic base include tertiary amine (such as triethylamine, Diisopropyl ethyl amine etc.)；Suitable inorganic base includes hydroxide (such as lithium hydroxide, hydroxide of alkali metal, alkaline-earth metal Sodium, potassium hydroxide etc.), carbonate (such as lithium carbonate, sodium carbonate, potassium carbonate), bicarbonate (such as sodium bicarbonate, saleratus Deng), phosphate (such as sodium phosphate, potassium phosphate), hydrophosphate (such as dibastic sodium phosphate, potassium hydrogen phosphate) etc..

In above method step (a), reaction dissolvent is selected from ethyl alcohol, isopropanol, triethylene glycol, the tert-butyl alcohol, isobutanol, diformazan Oxygroup ethane, toluene, n-butanol, glycerol, ethylene glycol, polyethylene glycol -200, polyethylene glycol-400, polyethylene glycol -600, poly- second Glycol -800, N-Methyl pyrrolidone, N,N-dimethylformamide, DMAC N,N' dimethyl acetamide, hexamethyl phosphoramide, dimethyl Sulfoxide, sulfolane, dioxane etc. or their mixture.

In above method step (a), reaction temperature is generally 70 DEG C to solvent boiling point.

In above method step (b), " diazo reagent " is selected from nitrous acid or nitrous acid ester etc., and wherein nitrous acid can It is prepared in situ by nitrite (such as sodium nitrite, potassium nitrite) and sour (such as hydrochloric acid, acetic acid), wherein nitrous acid ester includes Isoamyl nitrite etc..

In above method step (b), the molar ratio of VIII compound of formula and diazo reagent is generally 1:0.8 extremely 1:2。

In above method step (b), solvent is selected from or mixtures thereof toluene, acetic acid, water etc..

In above method step (b), reaction temperature is generally -15 DEG C~40 DEG C.

Above method step (a) or (b) in, the determination in reaction time can be used this field routine method carry out, such as adopt With TLC, HPLC monitoring etc..

Above method step (a) or (b) in, can further comprise right with the conventional methods in the field after the completion of reaction Product the post-processing such as is separated, is purified or being directly used in without further purification after separation in next step.

In above method step (a), VII compound of formula is the common intermediate for preparing ticagrelor, commercially viable purchase It obtains or is made by literature method；VI compound of formula or its sour addition product can be by the preparation method systems in following embodiments ?.

In one embodiment, the present invention provides the preparation method of a kind of VI compound of formula or its sour addition product, the party Method includes:

Ⅸ compound of formula or its sour addition product are obtained Ⅹ compound of formula through amido protecting by (I),

In formula Ⅹ, R₃For amino protecting group, R₄For hydrogen or and R₃It is used as amino protecting group together；

(II) reacts Ⅹ compound of formula with Ⅺ compound of formula to obtain Ⅻ compound of formula,

R in formula Ⅹ and Ⅻ₃And R₄It is defined as above, the R in formula Ⅺ₅For F, Cl, Br, I ,-OH or-OA, A is defined as above；

Ⅻ compound of formula removal amino protecting group is obtained VI compound of formula by (III),

R in formula Ⅻ₃And R₄It is defined as above；

It is (IV), optional, as needed by the sour addition product of VI compound of formula conversion VI compound of an accepted way of doing sth.

In above method step (I), Ⅸ compound of formula is the common intermediate for preparing ticagrelor, commercially viable purchase It obtains or is made by literature method.Acid in the sour addition product of Ⅸ compound of formula includes inorganic acid or organic acid；Wherein it is suitable for Inorganic acid be selected from hydrochloric acid, hydrobromic acid, hydroiodic acid, nitric acid, phosphoric acid or sulfuric acid etc.；Suitable organic acid is selected from L-TARTARIC ACID, two It is benzoyl-L-TARTARIC ACID, two pairs of toluyl groups-L-TARTARIC ACIDs, oxalic acid, acetic acid, trifluoroacetic acid, methanesulfonic acid, benzene sulfonic acid, right Toluenesulfonic acid etc..

In above method step (I), " amido protecting " is the ordinary skill in the art；Common amino protecting group packet Include benzyloxycarbonyl group (Cbz), tertbutyloxycarbonyl (Boc), tablet held before the breast by officials methoxycarbonyl group (Fmoc), allyloxycarbonyl (Alloc), trimethyl silicane second Oxygen carbonyl (Teoc), phthalyl (Pht), p-toluenesulfonyl (Tos), trifluoroacetyl group (Tfa), adjacent (to) nitrobenzene Sulfonyl (Ns), trityl (Trt), 2,4- dimethoxy-benzyl (Dmb), to methoxy-benzyl (Pmb) etc.；" the R₄With R₃ Together as amino protecting group " situation for example, when amino protecting group select phthalyl (Pht) when, R₄With R₃Generation together Table phthalyl.When selecting amino protecting group, preferred amino protecting group is is made when removing the amino protecting group Method is conducive to ehter bond in VI compound of formula and keeps stablizing.

In above method step (I), the acid of a certain amount of alkali neutralization reaction generation need to generally be added or by Ⅸ compound of formula The acid that sour addition product introduces.The alkali includes organic base or inorganic base；Wherein suitable organic base include tertiary amine (such as triethylamine, Diisopropyl ethyl amine etc.)；Suitable inorganic base includes hydroxide (such as lithium hydroxide, hydroxide of alkali metal, alkaline-earth metal Sodium, potassium hydroxide etc.), carbonate (such as lithium carbonate, sodium carbonate, potassium carbonate), bicarbonate (such as sodium bicarbonate, saleratus Deng), phosphate (such as sodium phosphate, potassium phosphate), hydrophosphate (such as dibastic sodium phosphate, potassium hydrogen phosphate) etc..

In above method step (II), Ⅺ compound of formula is commercially viable to be bought.

In above method step (II), a certain amount of alkali need to generally be added, the alkali includes organic base or inorganic base；Wherein Suitable organic base includes tertiary amine (such as triethylamine, diisopropyl ethyl amine), alcoholates (such as uncle of alkali metal, alkaline-earth metal Butanol lithium, sodium tert-butoxide, potassium tert-butoxide, tert-butyl alcohol magnesium etc.) etc.；Suitable inorganic base includes the hydrogenation of alkali metal, alkaline-earth metal Object (such as sodium hydride, hydrofining), hydroxide (such as lithium hydroxide, sodium hydroxide, the potassium hydroxide of alkali metal, alkaline-earth metal Deng), carbonate (such as lithium carbonate, sodium carbonate, potassium carbonate), bicarbonate (such as sodium bicarbonate, saleratus), phosphate (such as sodium phosphate, potassium phosphate), hydrophosphate (such as dibastic sodium phosphate, potassium hydrogen phosphate) etc..

In above method step (III), " the removal amino protecting group " is the ordinary skill in the art, is preferred beneficial for Ehter bond keeps stable method in VI compound of formula, for example removes amino protecting group using the methods of alkaline reagent, hydrogenolysis.

It can further comprise conventional with this field after the completion of reaction in above method step (I), (II) or (III) Method the post-processing such as is separated to product, is purified or being directly used in without further purification after separation in next step.

The present invention is on the basis of providing triazolopyrimidine derivative shown in preparation formula II, it is further provided in series Intermediate compounds therefor: III compound of formula, V compound of formula, Ⅻ compound of VI compound of formula, VIII compound of formula or formula, their structure And preferred compound is summarized in down:

R in formula₁For F, Cl, Br, I, OH or OA, wherein A is hydroxy activated base,

R in formula₁For F, Cl, Br, I, OH or OA, wherein A is hydroxy activated base,

R in formula₃For amino protecting group, R₄For.Hydrogen or and R₃It is used as amino protecting group together,

Purpose according to the present invention, the present invention provides a kind of II compounds of formula of crystalline state, and further, the present invention mentions A kind of crystal form for having supplied II compound of formula, is defined as crystal form A.

The feature of the powder x-ray diffraction map of II compound crystal form A of formula provided by the invention are as follows: 2 θ values be 4.4 ° ±0.2°、15.4°±0.2°、15.7°±0.2°、18.8°±0.2°、20.0°±0.2°、20.7°±0.2°、21.1°± 0.2 °, 26.2 ° ± 0.2 °, 26.6 ° ± 0.2 °, 27.5 ° of ± 0.2 ° of equipotentials be equipped with characteristic diffraction peak.

In one embodiment, the powder x-ray diffraction map of II compound crystal form A of formula provided by the invention Feature are as follows: 2 θ values be 4.4 ° ± 0.2 °, 10.3 ° ± 0.2 °, 15.4 ° ± 0.2 °, 15.7 ° ± 0.2 °, 17.3 ° ± 0.2 °, 18.8°±0.2°、20.0°±0.2°、20.7°±0.2°、21.1°±0.2°、21.7°±0.2°、26.2°±0.2°、26.6° ± 0.2 °, 27.5 ° ± 0.2 °, 28.2 ° of ± 0.2 ° of equipotentials be equipped with characteristic diffraction peak.

Further, the powder x-ray diffraction map that II compound crystal form A of formula of the present invention is indicated with 2 θ angles There is characteristic diffraction peak and relative intensity in following position:

In one embodiment, there is II compound crystal form A of formula provided by the invention powder X-ray-as shown in Figure 1 to penetrate The representative feature of ray diffraction diagram spectrum, i.e., the powder x-ray diffraction map of II compound crystal form A of formula provided by the invention are basic As shown in Figure 1.

II compound crystal form A of formula of the invention powder x-ray diffraction analysis be under environment temperature and ambient humidity, The source CuK α through Dutch Panaco X`Pert PRO type Powder X-ray DiffractometerWhat measurement was completed." environment temperature Degree " is usually 0~40 DEG C；" ambient humidity " is usually 30%~80% relative humidity.

In one embodiment, II compound crystal form A of formula provided by the invention has differential scanning as shown in Figure 2 Feature representated by calorimetry (DSC) map, i.e. its endotherm peak temperature are 150 DEG C ± 2 DEG C or 150 DEG C ± 1 DEG C.

In one embodiment, II compound crystal form A of formula provided by the invention has thermogravimetric analysis as shown in Figure 3 (TGA) feature representated by map, i.e., thermogravimetric analysis (TGA) map of II compound crystal form A of formula provided by the invention is substantially such as Shown in Fig. 3.

In one embodiment, II compound crystal form A mixture Chinese style of formula, II chemical combination of preparation provided by the invention Object crystal form A content (mass content) is generally higher than 70%, preferably greater than 80%, most preferably greater than 90%.

The present invention provides the preparation methods of II compound crystal form A of formula a kind of, this method comprises:

(1), II compound of formula is dissolved in esters solvent；

(2), solid is precipitated；

(3), divide isolated solid；

(4), optional, separated solid is dried.

In above method step (1), " esters solvent " is selected from ethyl acetate, methyl acetate, isopropyl acetate, acetic acid N-propyl, n-butyl acetate etc. or their mixed solvent, ethyl acetate, isopropyl acetate, n-propyl acetate or they Mixture.

In above method step (1), the selected solvent usage as unit of ml and II compound amount of formula as unit of g Ratio be generally 1:1 to 30:1；The mode that heating can be used is dissolved.

In above method step (1), the solution temperature of II compound of formula is generally room temperature to solvent boiling point.

In above method step (2), the mode of described " solid is precipitated " includes that solid is precipitated in cooling, addition anti-solvent is precipitated Solid, addition crystal seed precipitation solid etc. is precipitated after partial solvent is concentrated out in solid, these methods can be used alone can also group It closes and uses, can carry out, can also be carried out under static conditions under agitation.It is right at normal temperature that " anti-solvent " refers to The bad solvent of the dissolubility of II compound of formula, such as isooctane, n-hexane, normal heptane, petroleum ether or their mixture, In preferably isooctane, normal heptane.

In above method step (3), the mode of " separation " can be using suction filtration or centrifugation.

In above method step (4), the temperature of " drying " is generally 20~120 DEG C, preferably 40~100 DEG C；It can be with Constant pressure and dry can also be dried under reduced pressure.

In one embodiment, the preparation method of II compound crystal form A of formula of the invention includes:

(1), II compound of formula is dissolved in isopropyl acetate；

(2), cooling or addition anti-solvent crystallization；Wherein anti-solvent is selected from isooctane, n-hexane, normal heptane, petroleum ether etc. Or their mixture, preferably isooctane, normal heptane；

(3), divide isolated solid；

(4), optional, separated solid is dried；Drying temperature is generally 20~120 DEG C, preferably 40~100 ℃；It can also be dried under reduced pressure with constant pressure and dry.

Another object of the present invention is to provide brilliant containing II compound of formula, crystallization II compound of stance or II compound of formula The pharmaceutical composition of type A and by II compound of formula, crystallization II compound of stance or II compound crystal form A of formula be used for manufacturer's medication The application of object.

In order to realize the purpose, on the one hand the present invention provides a kind of II compound of formula comprising therapeutically effective amount, crystallization The pharmaceutical composition of II compound of stance or formula II compound crystal form A and pharmaceutic adjuvant.

On the other hand, the present invention provides II compound of formula, crystallization II compound of stance or II compound crystal form A of formula to make Standby anticoagulation, antithrombotic treat or prevent cerebral ischemia diseases or improve the application in the drug slept.Usually treatment is had Medicine is made in II compound of formula, crystallization II compound of stance or the II compound crystal form A of formula of effect amount and one or more pharmaceutic adjuvants Compositions or preparation, the pharmaceutical composition or preparation are prepared in a manner of well known in the art.

Aforementioned pharmaceutical compositions or preparation can be used as the drug of a kind of anticoagulation or antithrombotic, be mainly used for treating or preventing Thrombosis and its complication with coronary artery, the cerebrovascular or peripheral artery disease patient, it is particularly, acute for reducing The incidence of the thrombotic cardiovascular event of coronary syndrome patient.In addition, aforementioned pharmaceutical compositions or preparation can be used as A kind of drug for treating or preventing cerebral ischemia diseases (such as ischemic cerebral apoplexy is medium) or improving sleep.

Aforementioned pharmaceutical compositions or the dosage form of preparation include: tablet, capsule, pill, granule, syrup, powder, tongue Lower tablet, suspension, solution, injectable formulation, aerosol, dry powder doses, suppository, cream, ointment, gelling agent etc..It According to respective dosage form the characteristics of, administration route include oral, sublingual, parenterally (as intravenous injection, intramuscular injection, subcutaneously infuse Penetrate), transpulmonary/tracheae or percutaneous etc..

The dosage of above-mentioned composition or preparation according to conditions of patients property and seriousness, administration route and patient age, Weight etc. is adjusted, and general daily dose is in 1mg between 1g, and preferably 30mg is between 300mg；Daily can with single administration, It can also be with multiple dosing.

It can also include other suitable active constituents in aforementioned pharmaceutical compositions or preparation.

In one embodiment, pharmaceutical composition provided by the invention is oral solid formulation, preferred tablet or glue Wafer.The oral solid formulation except II compound of active constituent formula, crystallization II compound of stance or II compound crystal form A of formula in addition to, Also contain pharmaceutic adjuvant, the pharmaceutic adjuvant is the pharmaceutic adjuvant of this field routine, including filler, disintegrating agent, bonding Agent, lubricant etc..

The filler generally comprise mannitol, calcium monohydrogen phosphate, microcrystalline cellulose, pregelatinized starch, lactose, sucrose, Calcium sulfate, superfine silica gel powder etc., they, which can be used alone, to be used in mixed way.

The disintegrating agent generally comprises sodium carboxymethyl starch, sodium carboxymethylcellulose, cross-linked carboxymethyl cellulose sodium, crystallite Cellulose, crosslinked polyvinylpyrrolidone, polyvinylpyrrolidone, low-substituted hydroxypropyl cellulose, agar, calcium carbonate and carbonic acid Hydrogen sodium etc., they, which can be used alone, to be used in mixed way.

The adhesive generally comprises hydroxypropyl cellulose, hydroxypropyl methyl cellulose, polyethylene glycol, povidone, micro- Crystalline cellulose, starch slurry, water, ethanol solution of various concentration etc., they, which can be used alone, to be used in mixed way.

The lubricant generally comprises magnesium stearate, talcum powder, stearic acid, calcium stearate, palmitinic acid, alumina silicate, hard Acyl amine, solid polyethylene glycol etc., they, which can be used alone, to be used in mixed way.

If necessary, other auxiliary materials can also be added into aforementioned pharmaceutical compositions or preparation, as sweetener (such as Ah Si Patan, Steviosin etc.), colorant (such as ferrous oxide, titanium dioxide), stabilizer (such as vitamin E, thymol, sweet ammonia Acid etc.), surfactant (such as Sodium Laurylsulfate) etc..

The preparation of above-mentioned oral solid formulation can be according to the conventional method for preparing oral solid formulation in the art It carries out, such as: tablet can be prepared using wet granule compression tablet mode, and the modes systems such as wet granulation is encapsulated can be used in capsule It is standby.When the oral solid formulation is tablet, it further can be coated as needed, film coated tablet or sugar coated tablet is made Agent.Coated substrate includes cellulose family, crylic acid resin, carbohydrate, such as hydroxypropyl grade methylcellulose, sucrose, wherein may be used also Add plasticizer, antiplastering aid, opacifier.

Experiments have shown that II compound crystal form A activity of II compound of formula provided by the invention, crystallization II compound of stance or formula Suitable with ticagrelor, oral absorption is better than ticagrelor, metabolizable at ticagrelor in vivo, thus when extending effect Between, good material base is provided clinically to reduce administration number of times, improving compliance；Furthermore II compound of formula, crystalline state The physicochemical property of II compound crystal form A of II compound of formula or formula is conducive to the preparation of production, storage and preparation；In addition, the chemical combination Object not only has the function of anticoagulation, antithrombotic, but also can be used to treat or prevent cerebral ischemia diseases or improve sleep.

Detailed description of the invention

Fig. 1 is powder x-ray diffraction (PXRD) map of II compound crystal form A of formula.

Fig. 2 is differential scanning calorimetry (DSC) map of II compound crystal form A of formula.

Fig. 3 is thermogravimetric analysis (TGA) map of II compound crystal form A of formula.

Specific embodiment

Below in conjunction with embodiment, the invention will be further described, can make professional and technical personnel in the field more fully Understand the present invention, the range of but do not limit the invention in any way.

In embodiment¹H NMR and¹³C NMR test is using deuterated dimethyl sulfoxide as test solvent, with tetramethylsilane Make internal standard, is measured at room temperature with II 400MHz Nuclear Magnetic Resonance of Bruke AV-.

Mass spectrometric measurement is completed with Agilent Quadrupole LC/MS 6120B, ESI holotype in embodiment.

Powder x-ray diffraction analysis is by Dutch Panaco X`Pert PRO type Powder X-ray Diffractometer in embodiment Measurement, it is 4 ° -50 ° that test condition, which is with θ-θ configuration, scanning range, and step-length is 0.0130 °, continuous scanning.Testing light source is Copper target K α radiation；Voltage and current is respectively 40kV and 40mA.Method for making sample are as follows: take appropriate sample with spoon at ambient conditions Be placed in the groove of glass load sample piece, suitably rolled with glass slide, be evenly distributed on sample in load sample piece groove, then with carry glass Piece strikes off sample surfaces.Sample does not rotate in its own plane during test.

Differential scanning calorimetric analysis is surveyed by the DSC 214Polyma type differential scanning calorimeter of resistance to production of speeding in embodiment Fixed.Test condition are as follows: heating rate is 10 DEG C/min, and Range of measuring temp is 35 DEG C -220 DEG C.

Thermogravimetric analysis is measured by the resistance to TG 209F3 type thermogravimetric analyzer for speeding production in embodiment.Test condition are as follows: Heating rate be 10 DEG C/min, 35 DEG C -350 DEG C of Range of measuring temp.

1 2- of embodiment [[(3aR, 4S, 6R, 6aS) -6- amino tetrahydro -2,2- dimethyl -4H- cyclopenta -1,3- Two oxygroup -4- bases] oxygroup]-Cvclopropvlmethvl ethylether (VI compound of formula) preparation

Step I: N- [(3aS, 4R, 6S, 6aR)-tetrahydro -6- (2- hydroxyl-oxethyl) -2,2- dimethyl -4H- cyclopentadiene And two oxygroup -4- base of -1,3-]-benzyq carbamate (Ⅹ -1 compound of formula) preparation

By 2- [[(3aR, 4S, 6R, 6aS) -6- amino tetrahydro -2,2- dimethyl -4H- cyclopenta -1,3- dioxa - 4- yl] oxygroup]-ethyl alcohol L-TARTARIC ACID salt (the L-TARTARIC ACID salt of Ⅸ compound of formula) 106.06g, hexone 1500ml, potassium carbonate 131.61g and water 658ml mixing, are then added dropwise benzyl chloroformate (Cbz-Cl) at 25~35 DEG C 54.16g continues stir about 14 hours after dripping, and stops reaction, separates upper organic phase, water phase hexone 500ml back extraction, merges organic phase, is concentrated under reduced pressure, and concentrate is purified by silica gel column chromatography to obtain Ⅹ -1 compound of formula.

¹H NMR (400MHz, DMSO-d₆) δ: 1.215 (s, 3H)；1.359 (s, 3H)；(1.715-1.769 m, 1H)； (2.083-2.147 m, 1H)；(3.415-3.509 m, 4H)；(3.785-3.839 m, 2H)；(4.406-4.420 d, 1H)； (4.467-4.482 d, 1H)；(4.714-4.739 m, 1H)；5.027 (s, 2H)；(6.855-6.875 d, 1H)；7.293-7.372 (m, 5H).

(+)-ESI-MS:352.2。

Step II: N- [(3aS, 4R, 6S, 6aR)-tetrahydro -6- (2- cyclopropyl-methoxy base oxethyl) -2,2- dimethyl - Two oxygroup -4- base of 4H- cyclopenta -1,3-]-benzyq carbamate (Ⅻ -1 compound of formula) preparation

Under nitrogen protection, formula Ⅹ -1 compound 48.00g and n,N-Dimethylformamide 300ml are mixed, then -5~ Potassium tert-butoxide 18.37g is added portionwise at 5 DEG C, then bromomethyl cyclopropane (formula Ⅺ -1 is added dropwise after stir about 1 hour at -5~5 DEG C Compound) 22.14g and n,N-Dimethylformamide 15ml mixed solution, continue to be stirred to react about at -5~5 DEG C after adding 5 hours；Add water 200ml quenching reaction, then plus ethyl acetate 400ml is extracted, and separates organic phase, organic phase is successively through water The washing of 400ml × 3, saturated sodium-chloride water solution 200ml washing, anhydrous sodium sulfate is dry, is concentrated under reduced pressure, and concentrate is through silicagel column Chromatographic purifying obtains Ⅻ -1 compound of formula.

¹H NMR (400MHz, DMSO-d₆) δ: -0.023-0.012 (m, 2H)；0.276-0.319 (m, 2H)；0.803- 0.874 (m, 1H)；1.154 (s, 3H)；1.297 (s, 3H)；(1.679-1.714 m, 1H)；(1.998-2.061 m, 1H)； (3.039-3.180 m, 2H)；(3.428-3.525 m, 4H)；(3.743-3.809 m, 2H)；(4.350-4.366 d, 1H)； (4.415-4.431 d, 1H)；(4.912-4.986 m, 2H)；(6.561-6.582 d, 1H)；(7.254-7.324 m, 5H).

(+)-ESI-MS:406.2。

Step III: 2- [[(3aR, 4S, 6R, 6aS) -6- amino tetrahydro -2,2- dimethyl -4H- cyclopenta -1,3- two Oxygroup -4- base] oxygroup]-Cvclopropvlmethvl ethylether (VI compound of formula) preparation

Ⅻ -1 compound 29.00g, dehydrated alcohol 250ml and 10% palladium carbon 11.60g are mixed, hydrogen is passed through, 20~ Stir about 21 hours at 25 DEG C；Filtering, washs filter cake with dehydrated alcohol 50ml and is merged into filtrate, formula is concentrated under reduced pressure to obtain in filtrate VI compound.

(+)-ESI-MS:272.3.

2 2- of embodiment [{ (3aR, 4S, 6R, 6aS) -6- [the bromo- 5- of 7- (rosickyite base) -3H- [1,2,3] triazol [4,5- D] pyrimidin-3-yl]-2,2- dimethyl tetrahydro-3aH- cyclopenta [d] [1,3] Dioxol-4 -yl } oxygroup]-1- The preparation of Cvclopropvlmethvl ethylether (III -2 compound of formula)

Step a:2- [((3aR, 4S, 6R, 6aS) -6- { [the bromo- 2- of 5- amido -6- (rosickyite base) pyrimidine-4-yl] amino } - 2,2- dimethyl tetrahydro-3aH- cyclopenta [d] [1,3] Dioxol-4 -yl) oxygroup]-1- Cvclopropvlmethvl ethyl The preparation of ether (VIII -2 compound of formula)

VI compound 17.20g of formula, formula VII -1 compound 23.87g and N-Methyl pyrrolidone 67ml are mixed, are added three Ethamine 22.45g, under nitrogen protection at 86~93 DEG C stir about 6 hours；Water 70ml and isopropyl acetate 200ml is added to extract, point Organic phase out, water phase are stripped with isopropyl acetate 100ml, merge organic phase, organic phase is successively through the washing of water 150ml × 3, decompression Concentration, concentrate obtain VIII -2 compound of formula after being purified by silica gel column chromatography.

¹H NMR (400MHz, DMSO-d₆) δ: 0.113-0.150 (m, 2H)；0.407-0.453 (m, 2H)；0.940- 0.977 (m, 4H)；1.226 (s, 3H)；1.383 (s, 3H)；(1.597-1.689 m, 2H)；(1.794-1.850 m, 1H)； (2.244-2.309 m, 1H)；(2.957-2.993 m, 2H)；(3.229-3.246 d, 2H)；(3.498-3.601 m, 4H)； (3.865-3.886 t, 1H)；(4.260-4.302 m, 1H)；(4.508-4.549 m, 2H)；4.626 (s, 2H)；6.556-6.573 (d, 1H).

(+)-ESI-MS:517.2,519.2.

Step b:2- [{ (3aR, 4S, 6R, 6aS) -6- [the bromo- 5- of 7- (rosickyite base) -3H- [1,2,3] triazol [4,5-d] Pyrimidin-3-yl]-2,2- dimethyl tetrahydro-3aH- cyclopenta [d] [1,3] Dioxol-4 -yl } oxygroup]-1- ring The preparation of hydroxypropyl methyl ethylether (III -2 compound of formula)

VIII -2 compound 24.50g and toluene 245ml are mixed, then sequentially added at 0~10 DEG C acetic acid 15.61g and Sodium nitrite in aqueous solution (sodium nitrite 5.55g+ water 8.9ml) continues stir about 1 hour at 0~10 DEG C after adding；Carbon is added Sour aqueous solutions of potassium (potassium carbonate 17.00g+ water 94ml) quenching reaction, separates organic phase, obtains the toluene solution of III -2 compound of formula, It can be directly used for reacting in next step.

(+)-ESI-MS:528.2,530.2.

Embodiment 3 (1S, 2S, 3R, 5S) -3- [7- { [(1R, 2S) -2- (3,4- difluorophenyl)-cyclopropyl] amino } -5- (rosickyite base) -3H- [1,2,3] triazol [4,5-d] pyrimidin-3-yl] -5- (2- cyclopropyl-methoxy base oxethyl) -1,2- ring penta The preparation of alkane glycol (II compound of formula)

Step 1:2- ({ (3aR, 4S, 6R, 6aS) -6- { [7- { [(1R, 2S) -2- (3,4- difluorophenyl)-cyclopropyl] ammonia Base } -5- (rosickyite base) -3H- [1,2,3] triazol [4,5-d] pyrimidin-3-yl] -2,2- dimethyl tetrahydro -3aH- cyclopentadiene And [d] [1,3] Dioxol-4 -yl oxygroup]-1- Cvclopropvlmethvl ethylether (V compound of formula) preparation

By toluene solution, water 70ml and (1R, 2S) -2- (the 3,4- difluorobenzene of resulting III -2 compound of formula of embodiment 2 Base) cyclopropylamine R-MA salt (the R-MA salt of IV compound of formula) 15.18g mixing, then three are added dropwise at 0~10 DEG C Ethamine 11.95g, stir about 2 hours at 10~20 DEG C again after dripping off；Organic phase is separated, organic phase is successively through acetic acid aqueous solution The toluene solution that V compound of formula is obtained after (acetic acid 1.96g+ water 245ml) and saturated sodium-chloride water solution 250ml washing, can be direct For reacting in next step.

(+)-ESI-MS:617.3.

Step 2:(1S, 2S, 3R, 5S) -3- [7- { [(1R, 2S) -2- (3,4- difluorophenyl)-cyclopropyl] amino } -5- (rosickyite base) -3H- [1,2,3] triazol [4,5-d] pyrimidin-3-yl] -5- (2- cyclopropyl-methoxy base oxethyl) -1,2- ring penta The preparation of alkane glycol (II compound of formula)

By the toluene solution of V compound of step 1 gained formula and methanol hydrochloride solution (concentrated hydrochloric acid 39.4ml+ methanol 55ml) Mixing, stir about 4.5 hours at 10~15 DEG C；Add water 100ml, is about then 8 with triethylamine tune pH at 0~10 DEG C, point Organic phase out, water phase are stripped with isopropyl acetate 250ml × 2, merge organic phase, organic phase is successively through water 200ml washing, saturation Sodium-chloride water solution 200ml washing, anhydrous sodium sulfate is dry and is concentrated under reduced pressure, and obtains II compound of formula.

¹H NMR (400MHz, DMSO-d₆) δ: 0.127-0.163 (m, 2H)；0.417-0.462 (m, 2H)；0.807- 0.843and 0.941-1.006 (m, 4H)；(1.343-1.393 m, 1H)；1.468-1.579and 1.661-1.715 (m, 3H)；(2.011-2.081 m, 1H)；2.103-2.149and 2.253 (m, 1H)；(2.601-2.676 m, 1H)；2.821- 2.890 (m, 1H)；(2.916-2.985 m, 1H)；3.083-3.093and 3.129-3.178 (m, 1H)；3.232-3.249 (d, 2H)；(3.509-3.631 m, 4H)；(3.759-3.785 m, 1H)；3.950 (m, 1H)；(4.546-4.596 m, 1H)；4.941- 5.009 (q, 1H)；(5.031-5.041 d, 1H)；(5.099-5.115 d, 1H)；(7.066 m, 1H)；7.262-7.350 (m, 2H)；8.913-8.924and 9.326-9.336 (d, 1H).

¹³C NMR (100MHz, DMSO-d₆) δ: 3.30,10.96,13.42,15.44,22.77,24.48,32.82, 33.62,34.51,61.09,68.94,69.61,74.26,74.76,75.17,82.32,115.24,115.42,117.38, 117.55,123.25,123.28,123.31,123.66,139.37,139.68,139.72,139.74,139.78,146.99, 147.11,148.59,148.72,149.41,149.53,149.86,151.02,151.15,154.43,155.85,168.79, 169.65。

(+)-ESI-MS:577.3.

The preparation of 4 formula of embodiment, II compound crystal form A

II compound 2g of formula is dissolved in isopropyl acetate 6ml at 40~50 DEG C, isooctane 6ml, stirring is then added Crystallization about 4 hours, filtering, filter cake was dried under reduced pressure at 40~45 DEG C, obtained II compound crystal form A of formula.

The powder x-ray diffraction map surveyed is shown in Fig. 1, and measured value such as following table (takes relative intensity to be greater than 5% diffraction The corresponding measured value in peak):

Differential scanning calorimetry (DSC) map surveyed is shown in Fig. 2.Thermogravimetric analysis (TGA) map surveyed is shown in Fig. 3.It should manage Solution other types equipment may provide different temperature readings with above-mentioned condition is different from.Therefore, provided number It cannot function as absolute value.Skilled artisan will appreciate that the exact value of these temperature will by compound purity, example weight, plus The influence of hot rate and partial size.

The preparation of 5 formula of embodiment, II compound crystal form A

II compound 2g of formula is dissolved in isopropyl acetate 2ml at 50~55 DEG C, isooctane 4ml, stirring is then added Crystallization about 4 hours, filtering obtained crystal form A.

The preparation of 6 formula of embodiment, II compound crystal form A

II compound 2g of formula is dissolved at room temperature in isopropyl acetate 12ml, isooctane 20ml, stirring analysis is then added About 4 hours brilliant, filtering, filter cake is dried under reduced pressure at 70~75 DEG C, obtains crystal form A.

The preparation of 7 formula of embodiment, II compound crystal form A

II compound 2g of formula is dissolved in ethyl acetate 4ml at 40~50 DEG C, is subsequently cooled to -10~0 DEG C, stirring analysis It is about 2 hours brilliant, it filters, is dried under reduced pressure at 40~45 DEG C of filter cake, obtains crystal form A.

The preparation of 8 formula of embodiment, II compound crystal form A

II compound 2g of formula is dissolved in isopropyl acetate 20ml at 40~50 DEG C, normal heptane 40ml is then added, stirs It mixes crystallization about 4 hours, filters, filter cake is dried under reduced pressure at 40~45 DEG C, obtains crystal form A.

The preparation of 9 formula of embodiment, II compound crystal form A

II compound 2g of formula is dissolved in isopropyl acetate 4ml and ethyl acetate 2ml at 40~50 DEG C, is then added different Octane 8ml stirring and crystallizing about 4 hours, is filtered, is dried under reduced pressure at 90~100 DEG C of filter cake, obtains crystal form A.

10 formula of embodiment, II compound crystal form A stability test

The following are II compound crystal form A of formula to be tested under high temperature, super-humid conditions, and related substance and crystalline substance are carried out after 15 days Type detection.

Related substance is detected with HPLC method:

Chromatographic condition: with octadecylsilane chemically bonded silica (Agilent ZORBAX SB-C₁₈, 4.6 × 150mm, 1.8 μ It m) is stationary phase, with the NaH of 0.01mol/L₂PO₄Buffer solution (with phosphorus acid for adjusting pH to 3.5) is mobile phase A, acetonitrile is flowing Phase B, column temperature is 40 DEG C, flow velocity 1ml/min, Detection wavelength 242nm, according to following table gradient elution:

Time (min)	A% (V/V)	B% (V/V)
			0	80	20
20	55	45
			25	55	45
45	30	70
			50	30	70
55	80	20
			60	80	20

The preparation of mobile phase: taking sodium dihydrogen phosphate dihydrate 1.56g, and water 1000ml is added to dissolve, and adjusts pH3.5 with phosphoric acid, With 0.22 μm of membrane filtration up to mobile phase A；Taking chromatography acetonitrile is Mobile phase B.

Test solution: (being protected from light operation) takes this product about 20mg, accurately weighed, sets in 10ml measuring bottle, is dissolved with diluent And be diluted to scale, shake up to get.

Measuring method: it is tested according to high performance liquid chromatography (two V D of annex of Chinese Pharmacopoeia version in 2010).

Testing result is as follows:

The test result shows: II compound crystal form A of formula is with good stability.

The tablet of the II compound crystal form A containing formula of embodiment 11 and its preparation

Prescription:

Component	Content (mg/ piece)
		II compound crystal form A of formula	99.0
Mannitol	126.0
		Calcium phosphate dibasic dihydrate	63.0
Hydroxypropyl cellulose	9.0
		Sodium carboxymethyl starch	9.0
Magnesium stearate	3.0

Preparation: by II compound crystal form A of formula, the mannitol, calcium phosphate dibasic dihydrate, hydroxypropyl cellulose in upper table component Mixed with sodium carboxymethyl starch, dry with water wet granulation, whole grain is mixed with magnesium stearate, tabletting to get.

The Pharmacodynamics in vitro of 12 platelet aggregation-against of embodiment is tested

The present invention is by inhibiting ADP to induce the external of rabbit platelet aggregation test evaluation formula II compound and ticagrelor Activity.

It takes rabbit hind leg venous blood to be centrifuged, separates the blood plasma rich in blood platelet, be separately added into a series of formula II of solvent, concentration ADP induced platelet aggregation is added after 37 DEG C of common incubation 5min in compound or ticagrelor.5min is detected after ADP is added Interior maximum platelet aggregation rate (PAGm).Each group is all provided with 3 parallel pipes.

Grouping and sample-adding situation are as follows:

Solvent DMSO in blood platelet system final concentration of 0.1%, will not be to blood platelet body compared with blank control group System has an impact.

Group	Maximum aggregation rate (%)	Inhibiting rate (%)
			Negative control group	49.7±1.5	-
DMSO vehicle control group	49.0±0.6	-

II compound of formula of various concentration and the comparison of maximum aggregation rate and inhibiting rate in ticagrelor group 5min are as follows:

The test result shows: II compound of formula has the anti-platelet activity similar with ticagrelor in vitro.

The internal pharmacodynamics test of 13 platelet aggregation-against of embodiment

The present invention has rated the activity in vivo of formula II compound and ticagrelor by Beagle dog platelet aggregation test.

6 male Beagle dogs are randomly divided into 2 groups, according to equimolar than II chemical combination of orally administration formula respectively after overnight fasting Object and ticagrelor.It is cleaned 7 days after first administration, carries out the 2nd wheel administration.Hematometry corresponding index is periodically taken after administration.

0.5,1,1.5,2,4,8,24,30 each acquisition whole blood 2mL, 3.2% sodium citrate 1:9 are anti-before each medicine and after medicine It is solidifying.Low-speed centrifugal (300g) 5min produces platelet rich plasma (PRP).Remaining sample continues high speed centrifugation (3000g) 10min system Take platelet poor plasma (PPP).PRP is adjusted to platelet concentration with PPP as 2 × 10⁸/ mL detects maximum platelet aggregation rate, Calculate L-Arginine.

The test result shows that the L-Arginine of II compound of formula is auspicious lower than for lattice in 8 hours upon administration Lip river, but after 8 hours, it is higher than ticagrelor.

Pharmacokinetic trial in 14 body of embodiment

The present invention has rated the internal pharmacokinetics of II compound of formula and ticagrelor by the administration of SD rat oral.

16 SD rats, are randomly divided into 2 groups, every group 8 (half male and half female), give according to equimolar than respectively after overnight fasting Give II compound of formula, ticagrelor.Respectively at administration before and administration after 0.133 (8min), 0.25 (15min), 0.5 (30min), 1,2,4,8,12 and for 24 hours respectively acquisition 200 μ L of whole blood, anticoagulant heparin.In 4 DEG C, 3200rpm/min is centrifuged 10min separated plasma, inspection Survey the concentration of wherein formula II compound and ticagrelor.

Each group main pharmacokinetic parameters are as follows:

The test result shows:

(1), after II compound of formula or ticagrelor orally administration rat, there are gender differences, ground with ticagrelor original Situation in data after Oral Administration in Rats ticagrelor is consistent.

(2), a large amount of ticagrelor is detected in II compound group of formula, show that II compound of formula in vivo can be smoothly It is metabolized as ticagrelor.

(3), II compound group of formula or the administration of ticagrelor group equimolar, but the AUC of II compound group ticagrelor of formula, C_maxIt is apparently higher than ticagrelor group, since II compound of formula itself also has the drug activity similar with ticagrelor, If the drug activity object exposed amount of II compound group of formula will be apparent from being higher than ticagrelor group based on drug activity object.This shows The bioavilability of II compound of formula and its property metabolin is higher than ticagrelor.

(4), the Tmax of II compound group ticagrelor of formula than ticagrelor delay, when being conducive to extend drug effect Between.

The influence of 15 pairs of embodiment local rats with cerebral ischemia cerebral infarct volumes

(1), test material and method

Wistar rat, weight 250-280g.Perioperatively individually raising, room temperature keep 23-25 DEG C, have by oneself feed and into Water.TMCAO model is prepared according to the method for longa etc..Rat 10% chloral hydrate anesthesia (350mg/kg, i.p.), body temperature 37 ± 0.5 DEG C are maintained, dorsal position is fixed on operating table.Skin is cut along neck median line, carefully separates right carotid (CCA), external carotid artery (ECA), internal carotid (ICA).ECA is ligatured and is cut, is straightened with ICA in line.One is cut on ECA The round end silication nylon rope (being coated with 0.1% poly-D-lysine) of one root long 4.0cm, diameter 0.26mm are thus open by osculum It is inserted into ICA about 1.85-2.00cm, refers to rat brain prerolandic artery Rolando section start, blocks the supply of blood flow of arteria cerebri media.Ischemic 2 is small When after carefully extract nylon wire out, ligation ECA opening and suture operation notch, animal is put back to be filled 24 hours again in cage.

(2), test grouping and administration

Rat is grouped at random: model control group, water for injection (100ml/kg), II compound administration group of formula (25,50, 100mg/kg), oral administration when MCA blocking causes after ischemic 10 minutes.

(3), the measurement of cerebral infarct volume

After rat reperfusion injury 24 hours, broken end takes brain at once, and removal tractus olfactorius, cerebellum and low brain stem are coronal by its Be cut into 6 (first to the 5th 2mm/ piece, the 6th 4mm), be immediately placed in 5ml contain 1.5ml4%TTC and 0.1ml1MK₂HPO₄Solution in dyeing (37 DEG C, be protected from light) 20-30 minute, stirred therebetween every 5 minutes once.It is dyed through TTC Afterwards, normal tissue dye deeply takes on a red color, and infarction tissue is white.By every group of brain piece marshalling, preservation of taking pictures.Seek the stalk of calculation every Unleavened dough product, and finally superposition is converted into infarct volume.Infarct volume is indicated with the percentage of shared cerebral hemisphere, to eliminate brain The influence of oedema.

Cerebral infarct volume (%)=(operation contralateral hemisphere volume-operation side hemisphere non-infarcted portion volume)/operation pair The volume * 100% of side hemisphere

(4), test result

After ischemic Reperfu- sion 24 hours 2 hours, the cerebral infarct volume (%) of solvent control group is 33.8%.Sham-operation group does not have There is any cerebral infarction to occur.Other group of cerebral infarct volume result is as follows:

The test result shows: with solvent control group ratio, the oral administration of II compound of formula can be reduced significantly cerebral infarction Volume, i.e. II compound of formula have the function of good improvement symptoms of cerebral ischemia.

The influence that embodiment 16 acts on Sleep in Rats

(1), test material

Animal origin: Kun Ming mice, 18-22 grams, male.22 ± 2 DEG C of Bio Clean Room for Animal experiment temperature, relative humidity 50-70%, animal feeding material.

This test sets II compound dosage of formula as 25mg/kg, separately sets distilled water control group.

Sample treatment: it respectively takes sample 25mg to add distilled water to 20ml respectively, makes into uniform suspension, for examination.

(2), yellow Jackets above threshold dosage hypnosis test

Mouse is randomly divided into II compound administration group of control group and formula, and continuous gavage was filled to sample 30 days in the 30th day sample After stomach 15 minutes, it is injected intraperitoneally to the yellow Jackets of groups of animals 50mg/kg.b.w, injection volume 0.2ml/20g.b.w, with Mouse righting reflex loss was used as sleep judgment criteria up to 1 minute or more, observed to groups of animals in yellow Jackets 60 minutes Time for falling asleep and sleeping time.

The influence of II compound on animals weight of formula is as follows:

The test result shows: II compound group the weight of animals of formula compared with the control group, difference that there are no significant.

Above threshold the influence of the yellow Jackets inducing mouse sleeping time of dosage is as follows:

Group	Number of animals (only)	Time for falling asleep (is divided)	Sleeping time (divides)
				Control group	10	5.79±2.05	40.96±8.16
II compound of formula	10	5.78±1.88	55.16±7.80*

* P < 0.05 is compared with the control group (Analysis of variance)

The test result shows: II compound group animal of formula above threshold dose of sodium pentobarbitone induction under sleeping time with Control group is compared, and has significant difference.

(3), yellow Jackets sub-threshold dose hypnosis test

Mouse is randomly divided into II compound administration group of control group and formula, and continuous gavage was filled to sample 28 days in the 28th day sample After stomach 15 minutes, it is injected intraperitoneally to the yellow Jackets of groups of animals 30mg/kg.b.w, injection volume 0.2ml/20g.b.w, with Mouse righting reflex loss was used as sleep judgment criteria up to 1 minute or more, observed to groups of animals in yellow Jackets 25 minutes The number of animals slept.

The influence of the yellow Jackets inducing mouse sleep incidence of sub-threshold dose is as follows:

Group	Number of animals (only)	Sleep number of animals (only)	It sleeps incidence (%)
				Control group	10	2	20
II compound of formula	10	6*	60*

* P < 0.05 is compared with the control group (through Chi-square Test)

The test result shows: II compound group of formula sub-threshold dose yellow Jackets induction under sleep number of animals and sleep Dormancy incidence compared with the control group, has conspicuousness poor.

Above-mentioned test result shows: II compound of formula has the function of good improvement sleep.

Claims (10)

1. triazolopyrimidine derivative shown in a kind of formula II or its officinal salt,

2. the preparation method of triazolopyrimidine derivative shown in a kind of formula II or its officinal salt, this method comprises:

(1), III compound of formula is reacted into V compound of production with IV compound of formula or its sour addition product,

In formula III, R₁For F, Cl, Br, I, OH or OA, wherein A is hydroxy activated base；

(2), V compound of formula is deprotected to obtain II compound of formula,

(3), optional, salt is made in II compound of formula as needed.

3. it is used to prepare V compound of intermediate formula of II compound of formula described in claim 1,

4. II compound crystal form A of formula described in claim 1, powder x-ray diffraction map 2 θ values be 4.4 ° ± 0.2 °, 15.4°±0.2°、15.7°±0.2°、18.8°±0.2°、20.0°±0.2°、20.7°±0.2°、21.1°±0.2°、26.2° ± 0.2 °, 26.6 ° ± 0.2 °, 27.5 ° ± 0.2 ° of position have characteristic diffraction peak.

5. II compound crystal form A of formula according to claim 4, in powder x-ray diffraction map 2 θ values be 4.4 ° ± 0.2°、10.3°±0.2°、15.4°±0.2°、15.7°±0.2°、17.3°±0.2°、18.8°±0.2°、20.0°±0.2°、 20.7°±0.2°、21.1°±0.2°、21.7°±0.2°、26.2°±0.2°、26.6°±0.2°、27.5°±0.2°、28.2° There is characteristic diffraction peak in ± 0.2 ° of position.

6. II compound crystal form A of formula according to claim 5, powder x-ray diffraction figure has figure as shown in Figure 1 Spectrum.

7. the preparation method of II compound crystal form A of formula described in claim 4, this method comprises:

(1), II compound of formula is dissolved in esters solvent；

(2), solid is precipitated；

(3), divide isolated solid；

(4), optional, separated solid is dried.

8. preparation method according to claim 7, wherein esters solvent is selected from ethyl acetate, acetic acid first in step (1) Ester, isopropyl acetate, n-propyl acetate, n-butyl acetate or their mixed solvent；The mode that solid is precipitated in step (2) is selected From cooling precipitation solid, precipitation solid after anti-solvent is precipitated solid, partial solvent is concentrated out is added or crystal seed precipitation solid is added.

9. a kind of pharmaceutical composition, it includes II compound or pharmaceutically acceptable salt thereof of formula described in claim 1 of therapeutically effective amount, Or the described in any item II compound crystal form A of formula of claim 4~6 and pharmaceutic adjuvant of therapeutically effective amount.

10. II compound or pharmaceutically acceptable salt thereof of formula described in claim 1 or the described in any item formulas II of claim 4~6 are changed Object crystal form A is closed to prepare anticoagulation, antithrombotic, treat or prevent cerebral ischemia diseases or improving the application in the drug slept.