CN1118139A

CN1118139A - Fibrinogen receptor antagonists

Info

Publication number: CN1118139A
Application number: CN94191248A
Authority: CN
Inventors: D·A·克莱蒙; J·J·巴尔德温; N·利弗敦; B·阿斯基
Original assignee: Merck and Co Inc
Current assignee: Merck and Co Inc
Priority date: 1993-02-22
Filing date: 1994-02-22
Publication date: 1996-03-06
Also published as: NO953270D0; KR960700722A; FI953916A0; CA2155123A1; HUT71796A; BG99863A; WO1994018981A1; AU680240B2; NO953270L; NZ262664A; AU6246594A; JP3173792B2; SK102495A3; CZ210895A3; PL310386A1; EP0684823A1; FI953916A; JPH08507072A; EP0684823A4; HU9502028D0

Abstract

Compounds of the invention have formula (1) or (2), for example (3). The compounds have fibrinogen receptor antagonist activity.

Description

Title fibrinogen deceptor antagonists of the present invention

Background of the present invention

The present invention relates generally to regulate cytoadherence, and relate to and suppress Fibrinogen and other protein combine with hematoblastic, and relate to the hematoblastic gathering of inhibition, is the hematoblastic gathering to IIb/IIIa fibrinogen deceptor site specifically.Fibrinogen is a kind of glycoprotein that is present in the blood plasma, and it participates in platelet aggregation and participates in fibrinous formation.Platelet is the fragment that the cell sample removes nuclear, can find that it also participates in condensing of blood in all mammiferous blood.The interaction of known fiber proteinogen and IIb/IIIa acceptor site is essential for normal platelet function.

When blood vessel is damaged or is damaged by other origin cause of formation factors, platelet just adhere to ruined in subcutaneous (sub endothethial) surface.Subsequently, the platelet delivery of biologically active composition of this adhesion and gathering together.Gathering is that for example thrombin, epinephrine or ADP cause with specific combining of platelet membrane receptor by agonist.The stimulation of agonist causes the exposure of potential fibrinogen deceptor at platelet surface, and the combining of Fibrinogen and glycoprotein iib/iiia receptor complex.

People had attempted using natural product and synthetic peptide to measure the mechanism of adhesion and platelet aggregation.For example, Rouslahti and Pierschbacher are at Science, 238, the Fibronectin matter that is present in extracellular matrix and the blood has been described, for example fibronectin, vitronectin, osteopontin, collagen protein, thrombospondin, Fibrinogen and the von Willebrand factor among the 491-497 (1987).Described protein contains tripeptides arginine-glycine-aspartic acid (RGD) as its glycoprotein iib/iiia recognition site.These tripeptides that contain arginine-glycine-aspartic acid are integrated member's identification in plain (integrin) by relevant receptor on gang's structure at least, and described integration element is to have two heterodimer protein of striding the film subunit.The configuration that this author is illustrated in tripeptide sequence in each protein may be crucial for identification specificity.

Cheresh is in Proc.Nat ' l Acad.Sci, U.S.A., 84, the leading adhesion receptor of Arg-Gly-Asp of being expressed by human endothelial cell has been described among the 6471-6475 (1987), it structurally is similar to the IIb/IIIa complex on platelet, but on the antigen with function on different.This receptor directly is included in the endotheliocyte with Fibrinogen, the von Willebrand factor and vitronectin adhesion.

Pierschbacher and Rouslahti are in J.of Biol.Chem., 262, (36), hypothesis among the 17294-17298 (1987), the Arg-Gly-Asp sequence is exactly separately receptor identification and bonded enough signals, and so to make the configuration of tripeptide sequence be measurable.Made various synthetic peptides, this author concludes, has been influenced the interaction of receptor-ligand significantly by the three-dimensional chemical configuration to the enantiomer replacement of this sequence or the Arg-Gly-Asp that addition influenced.This author also illustrates, by form the decapeptide cyclisation that a disulphide bridges produces between non-terminal residue Pen and Cys, makes that the effect of this peptide aspect the adhesion of inhibition and fibronectin is extremely low.

At Proc.Nat ' l Acad.Sci.U.S.A.81, among the 5985-5988 (1984), identical author has described the tetrapeptide variant in the cell recognition site of fibronectin, and it is keeping promoting the activity of adhesion.Peptide class description with a tetrapeptide recognition site is in United States Patent (USP) 4,589, in 881 and 4,614,517.The great polypeptide fragment of being permitted in the cell land of fibronectin has the cytoadherence activity.For example referring to United States Patent (USP) 4,517,686,4,661,111 and United States Patent (USP) 4,578,079.

People such as Ruggeri, in Proc.Nat ' l Acad.Sci.U.S.A.83, explored a series of synthetic peptides that length is 16 residues that are designed among the 5708-5712 (1986), it contains and suppresses Fibrinogen and platelet is bonded, and the asparagicacid residue of the RGD RGD and the valine of ining succession.Also can be referring to people such as Koczewiak, Biochem.23,1767-1774 (1984); People such as Ginsberg, J.Biol.Chem., 260 (7), 3931-3936 (1985); With people such as Haverstick, Blood 66 (4), 946-952 (1985).Other inhibitor are disclosed in European Patent Application No. 275,748 and 298,820.

Many low molecular weight polypeptide factors obtain known the separation from snake venom.These factors obviously have the high-affinity to gp IIb/IIIa complex.For example, people such as Huang, J.Biol Chem., 262,16157-16163 (1987); People such as Huang, Biochemistry 28, and 661-666 (1989) has described the primary structure of venom trigramin, promptly contains 72 amino acid polypeptides of RGD subunit.Echistatin is another kind of venom, and it has high-affinity to the gpIIb/IIIa complex.This polypeptide contains 49 aminoacid, and has RGD subunit and various disulphide bridges.People such as Gan, J.Biol.Chem., 263,19827-19832 (1988).Also can be referring to people such as Dennis, Proc.Nat ' l Acad.Sci.USA, 87,2471-2475 (1989).Therefore yet these snake venom factors also have the high-affinity that other is comprised the adhesive protein acceptor member of vitronectin and fibronectin receptor, the gpIIb/IIIa complex are not had a selectivity.

Although known tripeptide sequence Arg-Gly-Asp is present in certain peptide species of the promotion cell interconnect function that can double or suppress fibronectin and vitronectin, tripeptides Arg-Gly-Asp has low activity.At present, for be coupled to other aminoacid on this sequence be how to influence binding specificity also have few understanding.Transfer Merck﹠amp; Co., the United States Patent (USP) 5,023,233 of Inc. discloses little ring six peptides, and it contains sequence A rg-Gly-Asp, and it is useful anticoagulant.United States Patent (USP) 5,037,808 disclose use indyl anticoagulant, and this inhibitor it is believed that it is that interaction by between antagonism Fibrinogen and/or extracellular matrix proteins and the platelet gpIIa/IIIa receptor is worked.United States Patent (USP) 5,037,808 disclose plan guanidine radicals peptide compounds, and it contains a kind of Asp residue of anticoagulant.Application PCT/US90/02746 has described use antibody-polypeptide conjugates, and wherein said polypeptide contains Arg-Gly-Asp (RGD) sequence.

Application PCT/US91/00564 discloses the use Macrocyclic peptides, and it contains the RGD that both sides are proline residue, and is anticoagulant.Application PCT/US90/03788 discloses little ring anticoagulant, and it is the synthetic ring-type pentapeptide that contains a thioether bond in tripeptide sequence Arg-Gly-Asp and the ring.Application PCT/US90/05367 (being disclosed on May 2nd, 1991) also discloses use peptide and false peptide, N-amidino groups-piperidines-3-carboxyl glycyl-L-aspartoyl-L-valine for example, and it can suppress platelet aggregation and thrombosis in the mammalian.The ol cpds of piperazinyl or acyclic derivatives in European Patent Application No. 91103462 discloses and can comprise.Transfer Merck﹠amp; Co., Inc. and be disclosed in the European Patent Application No. 91300179.8 on July 17th, 1991 and disclose the linear polypeptide fibrinogen deceptor antagonists.European Patent Application No. 90101404.3 discloses compound R ¹-A-(W) a-X-(CH ₂) b-(Y) c-B-Z-COOR, wherein R ¹Be guanidine radicals or amidino groups, and A and B are selected from specific mono-substituted aryl or heterocyclic radical.

Although it is known being considered to suppress to combine a large amount of chemical compounds of anticoagulant or peptide analogues with platelet by Fibrinogen, yet the invention provides new fibrinogen deceptor antagonists, described inhibitor has significant in conjunction with active, and be useful therefore, based on reason described herein.Many very serious diseases and imbalance comprise the too much complication of thrombosis that can cause interior thrombosis of blood vessel and embolus.Myocardial infarction, apoplexy, phlebitis and many other serious diseases need new, effective fibrinogen deceptor antagonists.

Summary The compounds of this invention of the present invention has the following formula structure:

Or For example: Described chemical compound has the fibrinogen deceptor antagonists activity.

Detailed description of the present invention

Or

Wherein Q is: Or Q contains 1,2 or 3 to be selected from the heteroatomic of N, O or S and to be unsubstituted or by R ⁸The 4-9 unit's monocycle or the bicyclo-ring system that replace; AB shares the adjacent carbon and the condensed ring system of nitrogen-atoms, wherein

A contains 1,2 or 3 heteroatomic 5,6 or 7 yuan of saturated or unsaturated ring that are selected from O, S or N;

B contains 1,2 or 3 heteroatomic 5,6 or 7 yuan of saturated or unsaturated ring that are selected from O, S or N; R ¹Be H, C _1-4Alkyl, N (R ⁸) ₂,-N (R ⁸) SO ₂R ⁷, NR ⁸CO ₂R ⁷, NR ⁸C (O) R ⁷, NR ⁸C (O) N (R ⁷) R ⁸, N (R ⁸) SO ₂N (R ⁷) R ⁸, N (R ⁸) SO ₂N (R ⁸) C (O) OR ⁷, C (O) N (R ⁷) ₂, or and R ⁶The cyclic group that forms, it is defined as follows; R ²Be H, C _1-4Alkyl, C _1-4Branched alkyl, C _1-4Alkylaryl, or aryl; R ⁴Be H, C _1-4Alkyl, C _1-4Branched alkyl, ring-type C _1-4Alkyl or C _1-4Alkenyl; R ⁵Be CH ,-CH (CH ₂) n, a key, probable back R ⁵Be contiguous N (R ⁴),

R ⁶Be COOH, CH ₂OH, C (O) NR ⁷) ₂, CO ₂R ⁹, tetrazolium, the acyl group sulfonamide, or

Or and R ¹The cyclic group that forms, it is defined as follows; R wherein ¹With R ⁶The cyclic group that forms is

Wherein Y=O or S; R ⁷Be H, replacement or unsubstituted side chain or straight chain C _1-4Alkyl, side chain or straight chain low-grade alkenyl, C _1-4The aryl of alkylaryl, replacement or contain 1,2 or 3 N, S or the heteroatomic 5 or 6 yuan of heteroaryls of O,

Wherein the alkyl of Qu Daiing is that hydroxyl replaces or C _1-4The alkyl that alkoxyl replaces, and the aryl that wherein replaces is replaced by one, two or three following groups: halogen, C _1-4Alkoxyl, hydroxyl or C _1-4Alkyl; R ⁸Be H, side chain or straight chain C _1-4Alkyl; R ⁹Be H, C _1-4Alkyl or aryl; N is 0-7; N ' is 0-3: and a is

An or key, and officinal salt.

In one embodiment, described chemical compound has the following formula structure:

Or Wherein Q is

Or

N=0-7; N '=0-3:R ⁴=H, C _1-4Alkyl, C _1-4Branched alkyl, ring-type C _1-4Alkyl or C _1-4Alkenyl; R ³=CH ,-CH (CH ₂) n or a key; R ²Be H, C _1-4Alkyl, C _1-4Branched alkyl, C _1-4Alkylaryl, or aryl; R ¹=H, C _1-4Alkyl, N (R ⁸) ₂,-N (R ⁸) SO ₂R ⁷, NR ⁸CO ₂R ⁷, NR ⁸C (O) R ⁷, NR ⁸C (O) N (R ⁷) R ⁸, N (R ⁸) SO ₂N (R ⁷) R ⁸, N (R ⁸) SO ₂N (R ⁸) C (O) OR ⁷, C (O) N (R ⁷) ₂, or and R ⁶The cyclic group that forms; R ⁶=COOH, CH ₂OH, C (O) NR ⁷) ₂, CO ₂R ⁹, tetrazolium, the acyl group sulfonamide, or

Or and R ¹The cyclic group that forms, wherein R ¹With R ⁶The cyclic group that forms is

Wherein Y=O or S; R ⁷=H, replacement or unsubstituted side chain or straight chain C _1-4Alkyl, side chain or straight chain low-grade alkenyl, C _1-4The aryl of alkylaryl, replacement or contain 1,2 or 3 N, S or the heteroatomic 5 or 6 yuan of heteroaryls of O,

Wherein the alkyl of Qu Daiing for replaced by hydroxyl or by C _1-4The alkyl that alkoxyl replaces, and the aryl that wherein replaces is replaced by one, two or three following groups: halogen, C _1-4Alkoxyl, hydroxyl or C _1-4Alkyl; R ⁸=H, side chain or straight chain C _1-4Alkyl; R ⁹=H, C _1-4Alkyl or aryl; A=

Or key A=contains 1,2 or 3 heteroatomic 5,6 or 7 yuan of saturated, fractional saturations or undersaturated ring that is selected from O, S or N; B=contains 1,2 or 3 heteroatomic 5,6 or 7 yuan of saturated, the fractional saturations or the undersaturated ring that are selected from O, S or N, and wherein A and B form the condensed ring system of a shared adjacent carbons and nitrogen-atoms.

In The compounds of this invention, the component with asymmetric center exists with racemic modification, racemic mixture and with single enantiomer and/or diastereomer.All isomeric form include in the present invention.

In this embodiment of a class, described chemical compound has the following formula structure:

AB is selected from following groups

Wherein V is N or CR ⁷And D is CH ₂, CH ₂-CH ₂, CH ₂C (R ⁷) ₂CH ₂, or Wherein X=N or CR ³, R wherein ³=CN, C (O) N (R ⁷) R ⁸, Or With

In the group in such embodiment, described chemical compound has the following formula structure: Wherein AB is

Wherein V is that N or CH and D are CH ₂-CH ₂Or CH ₂C (R ⁴) ₂CH ₂, in another kind of embodiment, described chemical compound has the following formula structure:

AB is selected from following groups Wherein V is N or CR ⁷With D be CH ₂, CH ₂-CH ₂, CH ₂C (R ⁷) ₂CH ₂, or Wherein X=N or CR ³,

R wherein ³=CN, C (O) N (R ⁷) R ⁸, Or

With

Y wherein ³Be O or H ₂, in a group of such embodiment, described an ancient type of spoon compound has the following formula structure: A=

Be selected from AB Wherein V is N or CR ⁷And D is CH ₂, CH ₂-CH ₂, CH ₂C (R ⁷) ₂CH ₂, or

With

Wherein X=N or CR ³, R wherein ³=CN, C (O) N (R ⁷) R ⁸, Or In another group of such embodiment, described chemical compound has the following formula structure: Key of a=; Be selected from AB

With

Y wherein ³Be O or H ₂

The instantiation of The compounds of this invention is the chemical compound that is selected from following compounds and officinal salt thereof:

With

Other examples of The compounds of this invention are:

With

Term " officinal salt " is meant the nontoxic salts of The compounds of this invention, and described salt is generally by preparing free alkali and the organic or inorganic acid reaction that suits.Representational salt comprises following salt: acetate, benzene sulfonate, benzoate, bicarbonate, disulfate, biatrate, borate, bromide, Ca-EDTA, camsilate, carbonate, chloride, clavulanate, citrate, dihydrochloride, edetate, ethanedisulphonate, Estolate, esilate, fumarate, gluceptate, gluconate, glutamate, Glu, bismuth glycolyl arsanilate salt, hexyl resorcin salt, breathe out amine, hydrobromate, hydrochlorate, Hydroxynaphthoate, iodide, different thiosulfate, lactate, lactobionate, laruate, malate, maleate, mandelate, mesylate, bromomethane, methyl nitrate, dimethyl sulfate, mucate, naphthalene sulfonate, nitrate, oleate, oxalates, embonate, palmitate, pantothenate (panthothenate), phosphate/phosphor acid hydrogen salt, Polygalacturonate, Salicylate, stearate, basic acetate, succinate, tannate, tartrate, 8-Chlorotheophyline, toluene fulfonate, triethiodide, valerate.

Term " medicine effective quantity " is meant biologically or the medicine of medical response or the amount of medicament that can cause tissue, system or animal, and this is research worker or the clinicist sought just.Term " anticoagulant " comprises heparin and warfarin.Term " thrombolytic agent " comprises streptokinase and the former activator of tissue plasminogen.Term " the anti-aggregating agent prepared therefrom of platelet " comprises for example aspirin, ticlopidine and dipyridamole.

Term " alkyl " is meant straight or branched alkane, olefine or alkynes.

Term " aryl " is meant 5-10 undersaturated monocycle of unit or bicyclic groups.

Term " heteroaryl " is meant and contains 1,2,3 or 4 heteroatomic aryl.

Term " hetero atom " is meant N, O or S.

Term " ring " is unless outside more specifically defining in addition, be meant the monocyclic or bicyclic saturated rings with 5-10 unit.

Term " heterocycle " is meant and contains 1,2,3 or 4 heteroatomic ring.

In The compounds of this invention, heteroaryl and heterocyclic radical contain and are no more than 2 O atoms or 2 S atoms.

Term " alkoxyl " comprises a moieties, and wherein alkyl as defined above.

Term " aralkyl " and " alkaryl " comprise a moieties, and wherein alkyl and comprises an aryl moiety as defined above, and wherein aryl as defined above.Wherein n can be respectively the C of the integer of 1-10 or 2-10 _O-nOr C _1-nDefinition be meant aralkyl or the unitary alkyl component of alkaryl.

Term " halogen " comprises fluorine, chlorine, iodine and bromine.

Term " oxygen base " is meant oxygen (O) atom, and term " oxo " is meant (=0).Term " sulfenyl " is meant a sulfur (S) atom.In the used standardized denomination, the end portion of defined side chain is at first described in the disclosure text, then describes the functional group adjacent with junction point.For example, by C _1-6The C that alkyl-carbonyl-amino replaces _1-5Alkyl is equal to following formula:

Alkyl.

In below the flow process and embodiment, all ingredients symbol has following meaning: BOC (Boc): tertbutyloxycarbonyl, Pd-C: palladium/activated-carbon catalyst, DMF: dimethyl formamide, DMSO: dimethyl sulfoxine, CBZ: benzyloxycarbonyl group, CH ₂Cl ₂: dichloromethane, CHCl ₃: chloroform, EtOH: ethanol, MeOH: methanol, EtOAc: ethyl acetate, HOAc: acetic acid, BOP: hexafluorophosphoric acid benzotriazole-1-base oxygen base three

(dimethylamino) Phosphonium EDC:1-(3-dimethylamino-propyl)-3-second

Base carbodiimides Oxone: permonosulphuric acid potassium LDA: diisopropylaminoethyl lithium DMA:N, accelerine, HOBT: hydroxybenzotriazole.Treatment

The compounds of this invention can be used for suppressing and the active relevant integration fibroin matter-complex function of cell attachment.They can be applied on one's body the patient who needs inhibition people or gathering of mammal platelet or adhesion.

Some The compounds of this invention is promptly eliminated from circulation, and is used in particular for anticoagulant.Therefore, these chemical compounds can be used in the surgical operation (artery transplantation, endarterectomy) of peripheral arterial and in the operation on vessels of heart, in these operations, the processing of tremulous pulse and organ and/or the interaction of platelet and artificial surfaces are caused platelet aggregation and consumption.Accumulative platelet can form thrombosis and thromboembolism (thromboemboli).Described chemical compound can be applied on one's body these surgery patients, with the formation of pre-preventing thrombosis and thromboembolism.

The compounds of this invention can be used with oral form (for example tablet, capsule (they include separately and continue to discharge or slow releasing preparation), pill, powder, granule, elixir, tincture, suspension, syrup and Emulsion).Equally, described chemical compound can be used with intravenous (bolus injection liquid or transfusion), intraperitoneal, subcutaneous, Sublingual, intranasal or intramuscular form, and all types of service are known for the those of ordinary skill of drug world.Required effectively and the chemical compound of avirulence consumption can be used as anti-aggregating agent prepared therefrom.

The compounds of this invention can be applied to need be by suppressing Fibrinogen on one's body combining of platelet membrane glycoprotein composite I Ib/IIIa receptor prevented thrombotic patient.Described chemical compound can be used in the surgical operation (artery transplantation, endarterectomy) of peripheral arterial and in the operation on vessels of heart, in these operations, the processing of tremulous pulse and organ and/or the interaction of platelet and artificial surfaces are caused platelet aggregation and consumption.Accumulative platelet can form thrombosis and thromboembolism (thromboem boli).Described chemical compound can be applied on one's body these surgery patients, with the formation of pre-preventing thrombosis and thromboembolism.

Extracorporeal circulation generally is used for operation on vessels of heart, so that to blood oxygenation.PA is on the surface of external contour.Adhesion is depended on the gpIIb/IIIa on the platelet membrane and is adsorbed onto interaction (people such as Gluszko, Amer.J.physiol., 252 (H), 615-621 (1987)) between the celloglobulin on contour surface.The platelet that discharges from artificial surfaces demonstrates the anastalsis that weakens.Can use The compounds of this invention with Film with Preventing Adhesion.

Other application of The compounds of this invention comprise during the prevention thrombolytic therapy and after platelet thrombus formation, thromboembolism and inaccessible again, and prevention is crown and the postangioplasty of other tremulous pulsies and the platelet thrombus formation behind the coronary bypass-forming operation, thromboembolism and inaccessible again.Described chemical compound also can be used to prevent myocardial infarction to form.

Use the dosage range of The compounds of this invention to select: the type, kind, age, body weight, sex and the medical condition that comprise the patient by various factors; The order of severity of disease to be treated; Route of administration; Patients " renal function and liver function; And used particular compound or its salt.Common experienced doctor or veterinary can easily determine and regulation prevention, resist or stop the required effective amount of drug of development of disease.

When being used for the described time spent of doing, the oral dose of The compounds of this invention is about 0.01mg/kg body weight/day (mg/kg/ days) extremely about 100mg/kg/ days, preferred 0.05～100mg/kg/ days, and most preferably 0.1-20mg/kg/ days.When intravenous was used, during the constant rate of speed infusion, most preferred dosage was about 1 to about 10 μ g/kg/ minute.Advantageously, the separate doses that The compounds of this invention can be divided into 2,3 or 4 every day is used.In addition, the preferred chemical compound of the present invention can be with the intranasal form by the local intranasal vector administration that suits of using, perhaps percutaneous approach, use is to percutaneous patch known to a person of ordinary skill in the art (patches) form administration, for with the transdermal delivery system form administration, if dosage is successive all the time in the administration process certainly, and can not be interrupted.

In the methods of the invention, this paper chemical compound described in detail can constitute active component, and they generally are to use with the suitable mutually blended form of medicinal diluent, excipient or carrier (being referred to as " carrier " material herein), described suitable medicinal diluent, excipient or carrier are suitably selected according to required form of medication (being oral tablet, capsule, elixir, syrup etc.), and consistent with the conventional medicine practice.

For example, for with tablet or Capsule form oral administration, described active medicine component can combine with oral, nontoxic, pharmaceutically useful, inert carrier, for example lactose, starch, sucrose, glucose, methylcellulose, magnesium stearate, dicalcium phosphate, calcium sulfate, mannitol, Sorbitol etc.; For with the liquid form oral administration, the oral drugs component can for example ethanol, glycerol, water etc. be used in combination with any oral, nontoxic, pharmaceutically useful inert carrier.In addition, if need or necessity, also binding agent, lubricant, disintegrating agent and the coloring agent that is fit to can be incorporated in this mixture.The binding agent that is fit to comprises starch, gelatin, natural saccharide (for example glucose or beta lactose), corn-sweetener, natural and rubber polymer class (for example arabic gum, Tragacanth or sodium alginate), carboxymethyl cellulose, Polyethylene Glycol, wax class etc.The lubricant that uses in these dosage forms comprises enuatrol, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, sodium chloride etc.Disintegrating agent (disintegrators) comprises starch methylcellulose, agar, bentonite, xanthan gum etc. without limitation.

The compounds of this invention also can be used with liposome medication system form, for example little monolayer capsule, big monolayer capsule and multilamellar capsule.The liposome class can for example cholesterol, stearmide or phosphatidylcholine form by various phospholipid.

The compounds of this invention also can come medication as independent carrier by using monoclonal antibody, and the The compounds of this invention molecule is coupled on these monoclonal antibodies.The compounds of this invention can also with the soluble polymer coupling as the drug target carrier.Described polymer can comprise polyvinylpyrrolidone, pyran co-polymer, poly-hydroxyl-propyl group-MAAm-phenol, poly-hydroxyl-ethyl-agedoite-phenol or the poly(ethylene oxide)-polylysine that is replaced by the palmityl residue.In addition, The compounds of this invention also can with the biodegradable polymer coupling of a class that is used to realize that controlled delivery of pharmaceutical agents discharges, for example copolymer of polylactic acid, polyglycolic acid, polylactic acid and polyglycolic acid, poly-epsilon-caprolactone, poly butyric, poe class, polyacetals, poly-dihydropyran class, polybutylcyanoacrylate class, and the crosslinked or amphipathic block copolymer of hydrogel.

The compounds of this invention also can be used jointly with suitable anticoagulant, described anticoagulant comprises anti-platelet agents, for example heparin, aspirin, warfarin, persantin and other are known suppresses chemical compound and the medicament that clot forms, and thrombolytic medicament for example plasminogen activator or streptokinase, to reach the beneficial effect of the various vascular lesions of treatment.

Noval chemical compound of the present invention is to prepare by the method among the following embodiment.Most preferred The compounds of this invention is any or all chemical compound that elaborates in these embodiments.Yet these chemical compounds should not be construed as formation and only are considered to scope of the present invention, and any bonded of described chemical compound or its part itself all can constitute a scope.The following example further illustrates the details of each The compounds of this invention of preparation.Those skilled in the art can understand easily that the condition of following preparation method and the known variant of technology can be used to prepare these chemical compounds.Unless otherwise noted, all temperature are Celsius temperature.

Except following preparation method, also pointed out the active embodiment of several external biologicals of the chemical compound in the scope of the present invention.For purposes of illustration, one is used to assess the active test of fibrinogen deceptor antagonists and is based in the inhibiting assessment of platelet that ADP-is stimulated.Gathering needs Fibrinogen to be attached to and occupies on the platelet fibrinogen deceptor site.The Fibrinogen binding inhibitors suppresses to assemble.In the platelet aggregation algoscopy that the inhibiting ADP-of the chemical compound that is used for determining that the present invention is claimed stimulates; human blood platelets is separated from fresh blood; pass through differential centrifugation; then in no bivalent ions, the Tyrode buffer agent (pH7.4) that contains 2% bovine serum albumin, carrying out gel filtration on the Sepharose 2B, and human blood platelets is being collected in acid citrate/dextrose.

Platelet aggregation is being measured on the Chronolog aggregometer under 37 ℃.Reactant mixture contains the human blood platelets (2 * 10 through gel filtration ⁸/ ml), Fibrinogen (100 micrograms/ml (μ g/ml)), Ca ²⁺(1mM) with chemical compound to be tried.Described gathering begins by adding 10mM ADP after 1 minute in other components of adding.Then, reaction was carried out 2 minutes at least.Accumulative inhibition degree expression is the percent of viewed accumulative speed under the condition of unrestraint agent.IC ₅₀Be with respect to no described chemical compound in the same old way, suppress to assemble the dosage of 50% particular compound.

In the following example, all temperature are Celsius temperature, unless otherwise noted.

The preparation of flow process A sulfonamide midbody compound:

Altheine-α-Ding sulfonamide (also claiming N-(normal-butyl-sulfonyl)-altheine) A-2

To contain altheine (6.45g, 48.9mmol) and NaOH (2.0g, the solution of 100ml 50% Han Shui diox 50.0mmol) is cooled to 0 ℃ in ice bath.In this rapid stirred mixture, alternately added in the clock time at 30 minutes NaOH (2.2g, 50ml aqueous solution 55.0mmol) and pure fourth sulfonic acid chloride (7.0ml, 53.9mmol).Under the reduced pressure reaction solution being concentrated into volume is 50ml, and cools off aqueous residue, with dense HCl acidify, and with ethyl acetate (3 * 100ml) extraction, organic extract Na ₂SO ₄Drying, being concentrated into volume is about 50ml, adds absolute ether (50ml), the white depositions of gained is separated with vacuum filtration process, obtains A-2, mp.154-155 ℃.

L-β-Boc-α-Ding sulfonamido-Beta-alanine (also claims 2 (S)-(normal-butyl sulfonamido)-3-(N-Boc-alanine) A-3

To contain NaOH (6.04g, 50ml H 151mmol) ₂O solution is cooled to 0 ℃, and the adding bromine (1.40ml, 26.9mmol).Gained solution stirred 5 minutes at 0 ℃, then, add immediately A-2 (5.23g, 20.7mmol) and NaOH (1.66g, 15ml H 41.4mmol) ₂O solution, and this mixture stirred 5 minutes at 0 ℃, be heated to 80 ℃ then, and under this temperature, kept 15 minutes.Solution is cooled to 25 ℃ then, and with 12N HCl (11ml) acidify, and stirs and stop until gas release.Make solution be alkalescence by adding 2N NaOH then, and add 20ml THF and connect Bis(tert-butoxycarbonyl)oxide (9.0g, 41.4mmol).Spend the night 25 ℃ of stirrings, THF is removed in decompression, and (2 * 50ml) extract alkaline water with ethyl acetate.Use 10% KHSO then ₄Make water be alkalescence, and (3 * 100ml) extract with ethyl acetate.The acid extract thing Na that merges ₂SO ₄Drying is filtered, and evaporation, obtains white solid A-3, mp.111-112 ℃.

2 (S)-(normal-butyl sulfonamido)-3-alanine (A-4)

(3.83g, 200ml ethyl acetate solution 11.8mmol) is cooled to 0 ℃, makes HCl gas bubbling pass through this solution 5 minutes with A-3.Then this solution is warmed to 25 ℃, and stirred 30 minutes, be evaporated to 50% of its volume then, with the dilution of 100ml ether.The gained white solid is collected by vacuum filtration, obtains solid, shaped A-4.

2 (S)-(normal-butyl sulfonamido)-3-alanine ethyl ester (A-5)

(1.0g, 50ml ethanol solution 3.8mmol) is saturated with HCl gas, then reflux 3.0h with A-4.Evaporating solvent obtains pure white solid A-5. ¹H?NMR(300MHz，CD ₃OD)δ?4.38(m，2H)；4.32(m，1H)；4.23(m，2H)；2.85(m，2H)；1.65-1.45(m，4H)；1.3(t，2H)；0.96(t，3H).

N-tosyl-altheine (A-6):

(10.0g 75.7mmol) adds secondary being equipped with in the 500ml round-bottomed flask of magnetic stirring bar and charging hopper with altheine.Add 1N sodium hydroxide (85ml, 1.1 equivalents).(15.88g 83.27mmol) is dissolved in the ethyl acetate (100ml) with paratoluensulfonyl chloride.This solution is joined in the reaction flask under vigorous stirring.1N sodium hydroxide (85ml, 1.1 equivalents) is placed charging hopper, under vigorous stirring, be added dropwise in the reaction bulb in then during 2 hours.Reactant mixture was stirred 2 hours under room temperature again.Organic layer and water layer are separated, and (2 * 50ml) wash water layer with ethyl acetate.With aqueous liquid cools to 0 ℃, use hydrochloric acid (dense) acidify then.Obtain white crystalline solid, use the hot water recrystallization, obtain A-6. ¹HNMR (300MHz DMSO-d ₆) δ 7.91 (d, J=8.79Hz, 1H); 7.64 (d, J=8.06Hz, 2H); 7.32 (s, d (eclipsed), J=8.06Hz, 3H); 6.87 (s, br, 1H); 4.03 (m, 1H); 3.32 (s, H ₂C); 2.49 (m.1H); 2.43 (d, d, J=7.08,15.38Hz, 1H); 2.35 (s, 3H); 2.21 (d, d, J=6.11,15.38Hz, 1H).

2 (S)-tosyl amino-3-alanine (A-7)

To contain NaOH (22.0g, 100ml H 550mmol) ₂O solution is cooled to 0 ℃, be added dropwise to bromine (5.03ml, 97.5mmol).Gained solution was stirred 10 minutes at 0 ℃, and (21.5g is 75.0mmol) with NaOH (6.68g, H 161mmol) for disposable then adding A-6 ₂Cold (0 ℃) solution of O (50ml)., reaction is heated to 80 ℃, and under this temperature, kept 30 minutes after 20 minutes 0 ℃ of stirring, then cooling.With dense HCl this cooled solution is transferred to pH=7, the gained white solid obtains A-7 after filtration. ¹H?NMR(300MHz，DMSO-d ₆)δ8.2-7.2(br，2H，(NH，COOH))；7.70(d，J＝8.18Hz，2H)；7.38(J＝8.18Hz，2H)；3.7-3.0(br，2H，(NH ₂，H ₂O))；3.12(q，J＝4.76Hz，1H)；2.99(d，d，J＝4.64，11.96Hz，1H)；2.79(d，d，J＝9.52，11.96Hz，1H)；2.36(s，3H).

The tert-butyl group-2 (S)-(tosyl amino)-3-alanine (A-8)

In the 1L pressure bottle, (5.0g 19.4mmol) is suspended in the diox (100ml) with A-7.This bottle is cooled to-15 ℃, and isobutene. (100ml) is condensed in Gai diox.Add dense H ₂SO ₄(5ml), and, at room temperature stirred 36 hours with the sealing of this bottle.Open bottle, the careful excessive isobutene. of emptying.This solution is with ethyl acetate (200ml) dilution, with 1N NaOH (200ml) washing.Organic layer Na ₂SO ₄Drying is filtered, and evaporation obtains the A-8 of white crystalline solid shape. ¹H?NMR(300MHz，CDCl ₃)δ7.68(d，J＝8.18Hz，2H)；7.35(d，J＝8.18Hz，2H)；3.85(m，H)；2.93-2.79(m，2H)；2.32(s，3H)；1.38(s，9H).

2-hydrazino pyridine-3-t-butyl formate (1-2)

(735mg, ethanol 3.44mmol) (5ml) solution is cooled to 0 ℃, and (2.75g 86mmol) handles with being dissolved in anhydrous hydrazine in the ethanol (5ml) with the 6-chloro-nicotinic acid tert-butyl ester.Make this mixture reach 25 ℃, stirred 20 hours, be warmed to 60 ℃ then, and under this temperature, kept 2 hours.This mixture is soluble in water, use ethyl acetate extraction.Na is used in organic moiety water and salt water washing ₂SO ₄Drying concentrates and obtains oily 1-2, and it is directly used in the next step.

[2,3-dihydro-3-oxo-1,4-triazol [4,3-a] pyridine-6-yl]-t-butyl formate (1-3)

1-2 is dissolved in the 35ml toluene, and (1.15g is in toluene 3.9mmol) (35ml) reflux solution slowly to join carbonic acid two (trichloromethyl) ester.This mixture further refluxed 1.2 hours, and cooling, was added to the water, and used ethyl acetate extraction.Organic moiety is through Na ₂SO ₄Drying concentrates, and through flash chromatography on silica gel method purification, obtains 1-3. ¹H?NMR(DMSO-d ₆，300MHz)：δ＝1.58(s，9H)；7.25(d，1H)；7.45(d，1H)；8.25(s，1H)；12.40(s，1H).

2-(N-CBZ-piperidin-4-yl) ethyl iodide (1-4)

With benzyl chloroformate (6.05ml, 42.5mmol) handle 4-(2-hydroxyethyl) piperidines (Aldrich) (5.0g, 38.7mmol), saturated NaHCO ₃(50ml) and CH ₂Cl ₂Mixture (150ml).After at room temperature stirring 2.5 hours, isolate organic layer, and use H ₂Na is used in O and salt water washing ₂SO ₄Drying is filtered then, concentrates, and obtains protected alcohol, is colorless oil. ¹H?NMR(300MHz，CDCl ₃)δ7.26(m，5H)；5.13(s，2H)；4.23(d，2H)；3.63(t，2H)；2.85(t，2H)；1.83(d，2H)；1.68(m，2H)；1.43(m，1H)；1.03(m，2H)；

Should protected alcohol (6.3g, 23.9mmol) and Ph ₃P (7.0g, 23.9mmol), iodine (6.06g, 23.9mmol) and imidazoles (1.95g 28.7mmol) mixes in benzene (100ml), and refluxed 2.5 hours.With this solution cooling, filtration, concentrate then.Residue obtains white solid 1-4 through silica gel chromatography purification (1: 1 ethyl acetate/hexane). ¹H?NMR(300MHz，CDCl ₃)δ7.26(m，5H)；5.14(s，2H)；4.23(d，2H)；3.86(m，4H)；1.85(d，2H)；1.68(m，2H)；1.45(m，1H)；1.03(m，2H).

3-[[(2,3-dihydro-3-oxo-[2-(N-CBZ-piperidin-4-yl) ethyl]-1,2,4-triazol [4,3-a] pyridine-6-yl] formic acid (1-5)

(350mg 1.6mmol), and is heated to 60 ℃, keeps under this temperature 20 hours to handle the 1-3 that is dissolved in the 20ml acetonitrile with powdered potassium carbonate (630mg).This mixture is cooled to 25 ℃, it is added to the water, and use ethyl acetate extraction.Organic moiety Na ₂SO ₄Drying concentrates, and through flash chromatography on silica gel method purification (hexane solution of 35% ethyl acetate), obtains required ester, is yellow oil.

This tert-butyl ester crude product is warmed to 25 ℃ then through handling down at 0 ℃ with 15ml dichloromethane and 15ml trifluoroacetic acid, keeps under this temperature 1.2 hours, and is converted into acid.This mixture vacuum concentration to doing, is added to the water, uses ethyl acetate extraction.Organic moiety Na ₂SO ₄Drying concentrates, and with ethylacetate/ether (25/1) crystallization, obtains 1-5, is buff powder. ¹H?NMR(DMSO-d ₆，300MHz：δ＝1.05(m，2H)；1.45(m，1H)；1.75(m，4H)；2.79(m，2H)；3.95(q，4H)；5.05(s，2H)；7.30(m，6H)；7.50(d，1H)；8.25(s，1H).

3-[[[(2,3-dihydro-3-oxo-2-[2-(N-CBZ-piperidin-4-yl) ethyl]-1,2,4-triazol [4,3-a] pyridine-6-yl] carbonyl] amino] propanoic acid (1-6)

With 1-5 (75mg, 0.18mmol) dimethyl formamide (1ml) solution use successively hydroxybenzotriazole (40mg, 0.26mmol), diisopropyl ethyl amine (87 μ l, 0.5mmol), 2-alanine carbethoxy hydrochloride (40mg, 0.26mmol) and EDC (50mg, 0.26mmol) processing.This mixture was stirred 15 hours at 25 ℃.This mixture is soluble in water, use ethyl acetate extraction.Na is used in organic moiety water and salt water washing ₂SO ₄Drying concentrates, and obtains required ester.

This ethyl ester crude product is dissolved in 5ml THF, the 5ml water, and handles with 0.37ml 1N LiOH aqueous solution.This mixture was stirred 3 hours at 25 ℃, mixture is soluble in water, use ethyl acetate extraction.Na is used in organic moiety water and salt water washing ₂SO ₄Drying concentrates, and obtains 1-6, mp201-203 ℃ (decomposition).

3-[[[(2, and 3-dihydro-3-oxo-2-(2-(piperidin-4-yl) ethyl]-1,2,4-triazol [4,3-a] pyridine-6-yl] carbonyl] amino] propanoic acid (1-7)

Under 0 ℃, (64mg, (107.0mg 0.533mmol) handles acetonitrile 0.129mmol) (15ml) solution, and reactant mixture was stirred 0.5 hour with the iodo trimethyl silyl with 1-6.Extracted with diethyl ether is used in the reactant mixture water treatment, uses the silica gel chromatography purification, uses EtOH/NH ₄OH/H ₂O (10/1/1) eluting concentrates and obtains white foam shape thing.Use alcohol crystal, obtain 1-7, be white solid, mp267-269 ℃.

2 (S)-[(6-butyl sulfonyl) amino]-3-[[[2,3-dihydro-3-oxo-2-[2-(N-CBZ-piperidin-4-yl) ethyl]-1,2,4-triazol [4,3-a] pyridine-6-yl] carbonyl] alanine (1-8)

Make 1-5 by the described method of 1-6 is coupled on the A-4, obtain 1-8. ¹H?NMR(300MHz，DMSO?d ₆)δ8.26(t，1H)，8.50(s，1H)；7.63(d，1H)；7.54(d，1H)；7.4-7.31(m，6H)；5.01(s，2H)；4.10(m，1H)；3.96(m，4H)；3.60(m，1H)；3.46(m，1H)；2.95(t，2H)；2.73(brm，2H)；1.71(m，2H)；1.53(m，2H)；1.46(m，1H)；1.01(m，2H)；0.83(t，3H).

2 (S)-[(normal-butyl sulfonyl) amino]-3-[[[2,3-dihydro-3-oxo-2-[2-(piperidin-4-yl) ethyl]-1,2,4-triazol [4,3-a] pyridine-6-yl] carbonyl] alanine (1-9)

Press the described method of 1-7, use trimethyl silyl iodine at CH ₃Handle 1-8 among the CN, obtain 1-9. ¹H?NMR(300MHz，D ₂O)δ8.10(s，IH)；7.43(d，1H)；7.15(d，1H)；4.00(m，4H)；3.60(d，2H)；3.31(m，2H)；3.23(d，2H)；3.93(m，2H)；3.81(t，2H)；1.93(d，2H)；1.80(m，2H)；1.6-1.23(m，7H)；0.85(t，3H).

Flow process 2

6-picoline-3-methyl formate (2-2)

To being equipped with Dropping funnel vertical (type) condenser and CaCl ₂Insert 6-methylnicotinic acid 2-1 (5g, 100ml absolute methanol solution 36.5mmol) in the 250ml three-neck flask of drying tube.Reaction solution is cooled to-15 ℃ in the ice acetone bath, and drips SOCl ₂(5ml, 69.1mmol).Then with vlil 3 hours, cooling afterwards, removal of solvent under reduced pressure.The white solid of the gained saturated NaHCO of 60ml ₃Handle, and use CH ₂Cl ₂(3 * 50ml) extractions.The extract Na that merges ₂SO ₄Drying is filtered and reduction vaporization.Gained grease is placed after crystallization, obtains white solid 2-2. ¹H NMR (300MHz, CDCl ₃) δ 9.05 (d, J=1.4Hz, 1H); (8.08 dd, J=1.4 and 6.8Hz); 7.19 (d, J=6.8Hz, 1H); 3.98 (s, 3H); 2.63 (s, 3H).

6-bromoethyl Nicotinicum Acidum methyl ester (2-3)

With 2-2 (10.6g, 71.5mmol) with NBS (12.73g, 71.5mmol), 100mg benzoyl peroxide and 200ml CCl ₄Mix, under inert atmosphere, refluxed together 18 hours.With reaction solution cooling, filtration, concentrated, obtain heavy-gravity orange, it through flash chromatography on silica gel method purification, with 20% ethyl acetate/hexane eluting, is obtained required pyridine radicals bromide 2-3. ¹H NMR (300MHz, CDCl ₃) δ 9.05 (d, J=1.4Hz,, IH); (8.08 dd, J=1.4 and 6.8Hz); 7.19 (d, J=6.8Hz, 1H); 5.38 (s, 2H); 3.98 (s, 3H).

6-[2-(N-Boc-piperidin-4-yl) ethylamino]-picoline-3-methyl formate (2-5)

In the 250ml flask, put into 2-3 (1.0g, 4.34mmol), 2-4 (2.16g, 10.0mmol) and K ₂CO ₃(0.66g is 4.4mmol) at the anhydrous CH of 100ml ₃Mixture among the CN, and refluxing 3 hours cools off then and filters.Concentrating under reduced pressure filtrate at the enterprising circumstances in which people get things ready for a trip spectrometry of silica gel purification, is used 10%CH ₃OH/EtOAc makes eluant, obtains 2-5, is yellow residue. ¹H NMR (CDCl ₃) δ 9.18 (d, J=1.4Hz, 1H); 8.15 (dd, J=1.4 and 6.8Hz, 1H); 7.39 (d, J=6.8Hz, 1H); 4.08 (brd, J=12Hz, 2H); 3.98 (s, 2H); 3.95 (s, 3H); 2.75 (eclipsed m, 6H); 1.78 (d, J=12, Hz, 2H); 1.5 (eclipsed m, 4H); 1.4 (s, 9H); 0.98 (m, 2H).

1-(chloro carbonyl)-2,3-dihydro-3-oxo-2-[[2-(N-Boc-piperidin-4-yl) ethyl] imidazo [1,5-a] pyridine-6-yl] methyl formate (2-6)

(480mg 1.27mmol) is dissolved in the 50ml toluene, adds N, and (645ml 4.08mmol), and is cooled to 0 ℃ with this solution to accelerine with 2-5.To wherein in 30 minutes, being added dropwise to triphosgene (1.13g, 15ml toluene solution 3.18mmol).Be heated to 100 ℃ with after this solution, kept under this temperature 1.5 hours, cooling then with 1N HCl, water and saline (each 50ml) washed twice, is used Na ₂SO ₄Drying, evaporation obtains the yellow crystalline solid 2-6 of 515mg. ¹H NMR (CDCl ₃) δ 8.82 (d, J=1.4Hz, 1H); 8.25 (d, J=6.8Hz, 1H); 7.91 (d, J=6.8Hz, 1H) (dd, J=1.4 and 6.8Hz, 1H; 4.08 (brd, J=12Hz, 2H); 3.98 (s, 2H); 3.95 (s, 3H); 2.75 (eclipsed m, 6H); 1.78 (d, J=12.Hz, 2H); 1.5 (eclipsed m, 4H); 1.4 (s, 9H); 0.98 (m, 2H).

1-[(N, the N-diethylamino) carbonyl]-2,3-dihydro-3-oxo-2-[[2-(N-Boc-piperidin-4-yl) ethyl]-imidazo [1,5-a] pyridine-6-yl] methyl formate (2-7)

(0.3g 0.64mmol) is dissolved in 100ml CH with 2-6 ₂Cl ₂In, (105mg is 0.97mmol) with the saturated NaHCO of 50ml to add diethylamine hydrochloride ₃Solution.This two-phase mixture was stirred 1 hour, isolate organic layer then, use 10% citric acid, use saline (50ml) washing then, use Na ₂SO ₄Drying, evaporation obtains 2-7. ¹H NMR (CDCl ₃) δ 8.45 (d, J=1.4Hz, 1H); 7.15 (dd, J=1.4 and 6.8Hz, 1H); 6.78 (d, J=6.8Hz, 1H); 4.08 (m, 4H); 3.95 (s, 3H); 3.51 (m, 4H); 2.75 (m, 3H); 1.78 (d, J=12.Hz, 2H); 1.5 (eclipsed m, 4H); 1.4 (s, 9H); 1.1 (t, 6H); 0.98 (m, 2H).

1-[[(N, the N-diethylamino) carbonyl]-2,3-dihydro-3-oxo-2-[2-(N-Boc-piperidin-4-yl) ethyl] imidazo [1,5-a] pyridine-6-yl] carbonyl] the alanine tert-butyl ester (2-8)

(315mg 0.64mmol) is dissolved in 10ml CH with 2-7 ₃Among the OH, add 15mlH ₂O and 0.725ml 1N NaOH, and mixture at room temperature stirred 3.5 hours.Organic solvent is removed in decompression, and CH is used in the citric acid acidify of aqueous residue ₂Cl ₂Extraction.Organic extract liquid H ₂Na is used in O, salt water washing then ₂SO ₄Drying is filtered, and evaporation obtains required acid. ¹H NMR (CDCl ₃) δ 8.32 (d, J=1.4Hz, 1H); 7.10 (dd ,=1.4 and 6.8Hz, 1H); 6.78 (d, J=6.8Hz, 1H); 4.08 (m, 4H); 3.51 (m, 4H); 2.75 (m, 3H); 1.78 (d, J=12.Hz, 2H); 1.5 (eclipsed m, 4H); 1.4 (s, 9H); 1.1 (t, 6H); 0.98 (m, 2H).

Should acid (105mg 0.215mmol) is dissolved in 10ml CH ₂Cl ₂In, add Et ₃(48ml 0.47mmol), is cooled to solution-10 ℃ to N.Then, (30ml 2.36mmol), stirs mixture 30 minutes at-10 ℃ to add isobutyl chlorocarbonate.To wherein add Beta-alanine tert-butyl ester hydrochlorate (58.7mg, 323mmol) and Et ₃N (32ml, 10ml CH 0.323mmol) ₂Cl ₂Solution, and solution is warmed to room temperature.Reaction solution is used Na with 10% citric acid, water and saline (each 10ml) washing ₂SO ₄Drying concentrates, and obtains 2-8 through chromatography purification. ¹H NMR (CDCl ₃) δ 8.16 (s, 1H); 7.02 (d, J=6.8Hz, 1H); 6.78 (d, J=6.8Hz, 1H); 6.63 (t, 5.6Hz, 1H); 4.08 (m, 4H); 3.51 (m, 6H); 2.75 (m, 2H); 2.45 (m, 2H); 1.78 (d, J=12.Hz, 2H); 1.5 (eclipsed m, 6H); 1.4 (s, 9H); 1.37 (s, 9H); 1.1 (t, 6H); 0.98 (m, 2H).

1-[(N, the N-diethylamino) carbonyl]-2,3-dihydro-3-oxo-2-[2-(piperidin-4-yl) ethyl] imidazo [1,5-a] pyridine-6-yl] carbonyl] propanoic acid (2-9)

With 2-8 (100mg 0.16mmol) is dissolved in the 20ml ethyl acetate, under 0 ℃ anhydrous HCl is reached 5 minutes in by solution, then with this mixture stirring at room 1 hour.Solvent evaporated under reduced pressure, residue is developed with ethyl acetate, filters, and obtains 2-9. ¹H NMR (DMSO-d ₆) δ 8.90 (brs, 1H); 8.60 (brs, 1H); 8.3 (s, 1H); 7.6 (t, 3H); 7.1 (d, 1H); 6.89 (d, 1H); 4.08 (m, 4H); 3.51 (m.6H); 2.75 (m, 2H); 2.45 (m, 2H); 1.78 (d, J=12.Hz, 2H); 1.5 (eclipsed m, 6H); 1.1 (t, 6H); 0.98 (m, 2H).

2-(N-Boc-piperidin-4-yl) ethanol (2-11)

With (Aldrich) (130g of 4-piperidines-2-ethanol (2-10), 1.0mol) be dissolved in the 700ml diox, be cooled to 0 ℃, (336ml is 1.0mol) with the Bis(tert-butoxycarbonyl)oxide (221.8g of company for usefulness 3N NaOH, 1.0mol) handle, remove ice bath, reactant mixture is stirred spend the night, concentrate, dilute with water is used extracted with diethyl ether.Combined ether layer is used the salt water washing, uses MgSO ₄Drying is filtered, and evaporation obtains 2-11.R _f=0.37 (1: 1 EtOAc/ hexane, painted) with 1,2,3-indantrione monohydrate. ¹H?NMR(300MHz，CDCl ₃)δ4.07(bs，2H)；3.7(bs，2H)；2.7(t.J＝12.5Hz，2H)；1.8-1.6(m，6H)；1.51(s，9H)；1.1(ddd，J＝4.3，12.5，12Hz，2H).

2-(N-Boc-piperidin-4-yl) ethyl iodide (2-12)

With 2-11 (10.42g 0.048mol) is dissolved in the 400ml benzene, at room temperature add imidazoles (4.66g, 0.068mol), triphenyl phasphine (15.24g, 0.05mol) and iodine (0.048mol).After 6 hours, filter reaction mixture, evaporated filtrate obtains dark-coloured residue.It through fast silica gel chromatogram method purification (with 10% EtOAc/ hexane eluting), is obtained 2-12, be yellow oil.

2-(N-Boc-piperidin-4-yl) ethylamine (2-4)

At room temperature, (27.9g, 0.082mol) (5.01g, 0.086mol), gained solution was 65 ℃ of heating 2 hours for middle adding Hydrazoic acid,sodium salt to the 2-12 that is dissolved in DMSO (400ml).Chilled reactant mixture with 2 * 100ml water, the extraction of 2 * 50ml saline, is used MgSO with 250ml EtOAc dilution then ₄Drying is removed and is desolvated, and obtains required azide, is light yellow oil, R _f=0.5 (silica gel, 70% acetone/hexane).

At room temperature, (19.3g is 0.076mol) at THF (400ml)/H to this azide ₂Disposable adding triphenyl phasphine in the mixture among the O (195ml) (80.0g, 0.305mol).At room temperature it was stirred 3 hours, under vacuum, remove organic solvent then.Use 10%KHSO ₄Solution is acidified to pH2 with residue, and extracts with 4 * 100ml EtOAc.Organic extract liquid is with 2 * 100ml 10%KHSO ₄Extraction merges water, with 2N NaOH pH is transferred to 10.This solution is with 4 * 200ml CH ₂Cl ₂Extraction with its merging, is used MgSO ₄Drying is removed and is desolvated, and obtains oily 2-4, R _f=0.3 (silica gel is used 10% CH ₃The CHCl of OH ₃/ NH ₃Eluant solution). ¹H NMR (300MHz, CDCl ₃) δ 4.05 (wide, 2H); 2.72 (t, J=7.2Hz, 2H); 2.62 (m, 2H); 1.64 (d, J=12.2Hz, 2H); 1.43 (s, 9H); 1.42-1.32 (m, 5H); 1.09 (m, 2H). flow process 3

1-(2-bromoethyl) pyrazoles-3,5-dioctyl phthalate dimethyl ester (3-2)

With anhydrous HCl gas treatment pyrazoles-3,5-dioctyl phthalate (75g, 1L absolute methanol solution 431mmol).Afterwards, continue to add HCl and reach 30 minutes, this solution is cooled to room temperature, placed 16 hours.Then with vlil 3 hours, cooling then, removal of solvent under reduced pressure.The gained white solid saturated NaHCO of 600ml ₃Handle, use CH ₂Cl ₂(3 * 500ml) extractions.The extract Na that merges ₂SO ₄Drying is filtered reduction vaporization.Gained white solid recrystallizing methanol adds absolute ether, obtains pyrazoles-3,5-dioctyl phthalate dimethyl ester (3-1a). ¹H?NMR(CDCl ₃)δ7.38(s，1H)；3.98(s，3H)；3.93(s，3H).

With this ester (5.0g, 150ml anhydrous acetonitrile K 27.2mmol) ₂CO ₃(5.2g, 40.0mmol) and glycol dibromide (25.0ml 291mmol) handles, and under argon atmospher the gained mixture heated is refluxed.After 25 minutes, the cooling reaction suspension filters, and filtrate evaporated under reduced pressure is extremely done, and placed high-vacuum tube to reach 12 hours.Gained white solid hexane recrystallization obtains white solid 3-2.

¹H?NMR(CDCl ₃)δ7.38(s，1H)；5.03(t，J＝8.2Hz，2H)；3.98(s，3H)；

3.93(s，3H)；3.75(t，J＝8.5Hz，2H).

[4,5,6,7-tetrahydrochysene-4-oxo-5-[2-(N-Boc-piperidin-4-yl) ethyl] pyrazolo [1,5-a] pyrazine-2-yl] methyl formate (3-3)

With 3-2 (14.0g, 48.0mmol), diisopropyl ethyl amine (25ml, 144mmol), Boc-4-amino-ethyl piperidines (12.0g, 52.6mmol) and potassium iodide (2.39g, 250ml CH 0.3mmol) ₃CN solution is at N ₂Refluxed 4.5 hours in the atmosphere, cooling is then filtered, and reduction vaporization.Gained yellow residue silica gel chromatography purification is made eluant with EtOAc, obtains 3-3, is the canescence crystalline solid. ¹H?NMR(300MHz，CDCl ₃)δ7.15(s，1H)；4.29(t，J＝7.0Hz，2H)；3.93(brd，J＝12Hz，2H)；3.76(s，3H)；3.61(t，J＝5.3Hz，2H)；3.42(t，J＝7.3Hz，2H)；2.65(t，J＝7.6Hz，2H)；1.55(d，J＝12.5Hz，2H)；1.38(m，2H)；1.33-1.25(m，1H)；1.27(s，9H)；1.01(m，2H).

To contain LiOH (14.05mg, 10ml H 0.335mmol) ₂O solution joins 3-3 (90.81mg, 10ml CH 0.223mmol) ₃In the OH solution, and mixture heated to 60 ℃ kept under this temperature 2.5 hours, cooling then, CH is removed in decompression ₃OH.Last water is used CH with 10% aqueous citric acid solution acidify ₂Cl ₂(2 * 50ml) extractions.The organic extract liquid Na that merges ₂SO ₄Drying, evaporation obtains required acid then, is white solid. ¹H?NMR(300MHz，CDCl ₃)δ7.43(s，1H)；4.48(t，J＝7.0Hz，2H)；4.01(brd，J＝12Hz，2H)；3.77(t，J＝5.3Hz，2H)；3.51(t，J＝7.3Hz，2H)；2.71(t，J＝8.3Hz，2H)；1.72(d，J＝12.5Hz，2H)；1.53(m，2H)；1.42-1.37(m，1H)；1.35(s，9H)；1.10(m，2H).

2 (S)-[(normal-butyl sulfonyl) amino-3-[[[4,5,6,7-tetrahydrochysene-4-oxo-5-[2-(N-Boc-piperidin-4-yl) ethyl] pyrazolo [1,5-a] pyrazine-2-yl] carbonyl] alanine (3-4)

With isobutyl chlorocarbonate (1.75ml, 13.35mmol) join contain this acid (4.98g, 12.72mmol) and N-methylmorpholine (1.53ml is in cold (0 ℃) solution of 100ml THF 14.00mmol).Under dry nitrogen atmosphere, stir this mixture.React after 1 hour, analyze sample aliquot with HPLC, the completeness that shows reaction is＞90%.Remove by filter N-methylmorpholine-HCl, with the filtrate impouring contain A-4 (4.30g, 16.54mmol), the isopropyl ethylamine (4.27ml, 33.10mmol), THF (60ml) and H ₂In the solution of O (20ml).Reduce pressure then and remove THF from reaction solution, remaining aqueous is partly used saturated KHSO ₄Acidify is with ethyl acetate (3 * 200ml) extractions.The extract Na that merges ₂SO ₄Drying is filtered and is concentrated, and obtains red oil, forms white solid 3-4 thus. ¹H NMR (DMSO-d ₆) δ 8.31 (t, J=6Hz, 1H); 7.62 (d, J=8.5Hz, 1H); 7.01 (s, 1H); 4.43 (t, J=6.6Hz, 2H); 4.11 (m, 1H); 3.92 (d, J=12Hz, 2H); 3.80 (t, J=6.6Hz, 2H); 3.51 (t, J=7.3Hz, 2H); 3.65 (m, 2H); 3.51 (t, J 6.8Hz, 2H); 2.96 (t, J=7.2Hz, 2H); 1.70 (d, J=11Hz, 2H); 1.53 (m, 2H); 1.60-1.49 (eclipsed m, 5H); 1.40 (s, 9H); 1.28 (q, J=7.1Hz, 2H); 1.05 (m, 2H); 0.79 (t, J=7.1Hz, 3H).

2 (S)-[(normal-butyl sulfonyl) amino]-3-[[[4,5,6,7-tetrahydrochysene-4-oxo-5-[2-(piperidin-4-yl) ethyl] pyrazolo [1,5-a] pyrazine-2-yl] carbonyl] alanine mono-hydrochloric salts (3-5)

(278mg, 30ml ethyl acetate solution 0.437mmol) is cooled to 0 ℃, and HCl gas bubbling was fed 3 minutes with 3-4.Reactant mixture is warmed to room temperature, stirred 30 minutes, on rotary evaporator, be evaporated to dried then.Made white solid filters, and uses P with ethanol/water (90: 10) recrystallization ₂O ₅Vacuum drying obtains white solid 3-5. ¹H NMR (DMSO-d ₆) δ 8.95 (brs, 1H); 8.33 (t, J=5.7Hz, 1H); 7.64 (d, J=9Hz, 1H); 7.02 (s, 1H); 4.35 (t, J=5.1Hz, 2H); 4.10 (m, 1H); 3.81 (t, J=5.2Hz, 2H); 3.6-3.4 (m, 4H); 3.21 (d, J=10.5Hz, 2H); 2.95 (t, J=7.8Hz, 2H); 2.81 (brm, 2H); 1.96 (d, J=11Hz, 2H); 1.62-1.2 (eclipsed multiplet, 9H); 0.80 (t, J=7.3Hz, 2H). flow process 4

2 (S)-N-benzyloxycarbonyl aminos-3-alanine methyl ester hydrochloride (4-1a)

Commercially available 2 (S)-N-benzyloxycarbonyl aminos-3-alanine (Fluka) was refluxed 2.5 hours in the HCl methanol solution, evaporation then, residue methanol crystallization obtains white solid 4-1a. ¹H?NMR(300MHz，DMSO-d ₆)δ7.63(m，5H)；5.93(d，1H)；5.15(s，2H)；4.56(m，1H)；3.95-3.83(m，2H)；3.73(s，2H).

2 (S)-[(CBZ) amino]-3-[[[4,5,6,7-tetrahydrochysene-4-oxo-5-[2-(N-Boc-piperidin-4-yl) ethyl] pyrazolo [1,5-a] pyrazine-2-yl] carbonyl] amino] methyl propionate (4-2)

At N ₂Under the atmosphere at room temperature, with 3-3 (5.6g, 14.0mmol), N α-Cbz-L-2,3-diaminopropionic acid methyl ester hydrochloride (4-1a) (4.5g, 15.5mmol), HOBT (2.37g, 15.5mmol) and Et ₃(4.1ml, 65ml anhydrous DMF solution 29.5mmol) stirred 48 hours N.DMF is removed in decompression, and residue is dissolved in the 700ml ethyl acetate, uses saturated NaHCO ₃Solution, H ₂O, 10% citric acid, H ₂(Na is used in each 1 * 100ml) washing successively for O and saline ₂SO ₄Drying is filtered, and evaporation.Gained clear glass shape thing is used 3%CH through the silica gel chromatography purification ₃OH/CH ₂Cl ₂Make eluant, obtain the pure 4-2 of white solid. ¹H?NMR(CDCl ₃)δ7.43(m，5H)；7.35(s，1H)；7.18(t，J＝6.5Hz，1H)；5.98(d，J＝6.8Hz，1H)；5.09(s，2H)；4.59(m，1H)；4.38(m，2H)；4.10(brd，J＝12Hz，2H)；3.8(s，3H)；3.73(t，J＝5.3Hz，2H)；2.71(t，J＝8.3Hz，2H)；1.72(d，J＝12.5Hz，2H)；1.53(m，2H)；1.42-1.37(m，1H)；1.35(s，9H)；1.10(m，2H).

2 (S)-amino-3-[[[4,5,6,7-tetrahydrochysene-4-oxo-5-[2-(N-Boc-piperidin-4-yl) ethyl] pyrazolo [1,5-a] pyrazine-2-yl] carbonyl] amino] methyl propionate (4-3)

To 4-2 (6.3g, 700ml CH 10.26mmol) ₃Add 650mg 10%Pd/C in the mixture of OH, with the gained mixture at 1 H ₂Stirred 48 hours in the air pressure.Remove catalyst with diatomite filtration, concentrated filtrate obtains flint glass shape thing, and it is used Et ₂The O development is filtered and is obtained white solid 4-3. ¹H?NMR(300MHz，CDCl ₃)δ7.43(m，5H)；7.35(s，1H)；7.18(t，J＝6.5Hz，1H)；5.98(d，J＝6.8Hz，1H)；5.09(s，2H)；4.59(m，1H)；4.38(m，2H)；4.10(brd，J＝12Hz，2H)；3.81(s，3H)；3.73(t，J＝5.3Hz，2H)；2.71(t，J＝8.3Hz，2H)；1.72(d，J＝12.5Hz，2H)；1.53(m，2H)；1.42-1.37(m，1H)；1.35(s，9H)；1.10(m，2H).

2 (S)-(acetylamino)-3-[[[4,5,6,7-tetrahydrochysene-4-oxo-5-[2-(N-Boc-piperidin-4-yl) ethyl] pyrazolo [1,5-a] pyrazine-2-yl] carbonyl] amino]-methyl propionate (4-4)

(70ml, (350mg is in cold (0 ℃) solution of 10ml THF 0.69mmol) 0.76mmol) to join 4-3 with acetic anhydride.Make gained solution be warmed to room temperature, and stirred 18 hours, concentrate then, residue is dissolved in the 50ml ethyl acetate, uses NaHCO successively ₃, H ₂O, 10%/KHSO ₄, H ₂O and saline (each 25ml) washing.Organic layer Na ₂SO ₄Drying, evaporation obtains colourless residue, and it through the silica gel chromatography purification, is used 3%CH ₃OH/CH ₂Cl ₂Eluting obtains white solid 4-4. ¹H?NMR(CDCl ₃)δ7.29(s，1H)7.24(t，J＝6.4Hz，1H)；6.81(d，J＝7.6Hz，1H)；4.79(m，1H)；4.38(m，2H)；4.10(brd，J＝12Hz，2H)；3.81(s.3H)；3.80(m，2H)；3.73(t，J＝5.3Hz，2H)；2.71(t，J＝8.3Hz，2H)；2.01(s，3H)；1.72(d，J＝12.5Hz，2H)；1.57(m，2H)；1.42-1.37(m，1H)：1.37(s，9H)；1.09(m，2H).

2 (S)-(acetylamino)-3-[[[4,5,6,7-tetrahydrochysene-4-oxo-5-[2-(piperidin-4-yl) ethyl] pyrazolo [1,5-a] pyrazine-2-yl] carbonyl] alanine (4-5)

With 4-4 (203mg, 0.38mmol), 1N LiOH (0.76ml, 0.76mmol), H ₂O, CH ₃The solution of OH and THF (each 5ml) at room temperature stirs and spends the night.Organic solvent, surplus solution 25ml H are removed in decompression ₂10%KHSO is used in the O dilution ₄Make solution be acid, use ethyl acetate extraction.Ethyl acetate layer H ₂Na is used in O and salt water washing ₂SO ₄Drying is filtered, and evaporation obtains required acid. ¹H?NMR(CDCl ₃)δ7.93(br，1H)；7.81(br，1H)；7.29(s，1H)；4.79(m，1H)；4.348(m，2H)；4.10(brd，J＝12Hz，2H)；3.80(brm，2H)；3.73(br，t，2H)；2.71(t，J＝8.3Hz，2H)；2.11(s，3H)；1.72(d，J＝12.5Hz，2H)；1.57(m，2H)；1.42-1.37(m，1H)；1.37(s，9H)；1.12(m，2H).

Should acid (169mg 32.8mmol) is dissolved in the 50ml ethyl acetate, and is cooled to 0 ℃, handles 30 minutes with anhydrous HCl.

Solvent removed in vacuo, residue is used P with absolute ether development, filtration ₂O ₅Drying obtains 4-5, is white solid, mp.150-156 ℃.

2 (S)-[(Cbz-amino)]-3-[[[4,5,6,7-tetrahydrochysene-4-oxo-5-[2-(N-Cbz-piperidin-4-yl) ethyl] pyrazolo [1,5-a] pyrazine-2-yl] carbonyl] alanine (4-6)

Use makes 4-3 be coupled to N α-CBZ-L-2 to the described method of 3-4,3-diaminourea-propanoic acid (Fluka) (4-1) on, obtain 4-6, the products of promptly two protections.

2 (S)-amino-3-[[[4,5,6,7-tetrahydrochysene-4-oxo-5-[2-(piperidin-4-yl) ethyl] pyrazolo [1,5-a] pyrazine-2-yl] carbonyl] alanine

In the presence of Pd/C, use H ₂Handle 4-6, obtain required acid, mp.157 ℃.The Boc group is removed in the usual way with HCl/EtOAc then, obtains pure 4-7, mp.195-198 ℃.

2 (S)-[(normal-butyl sulfonamido)]-3-[[[4,5,6,7-tetrahydrochysene-4-oxo-5-[, 2-(N-CBZ-piperidin-4-yl) ethyl] pyrazolo [1,5-a] pyrazine-2-yl] carboxyl] alanine methyl ester (4-8)

With 4-3 (0.30g, 0.61mmol), the normal-butyl sulfonic acid chloride (0.16g, 0.91mmol) and the 50ml THF solution of N-methylmorpholine at room temperature stirred 1.2 hours.Solvent evaporated under reduced pressure is dissolved in CH with gained grease ₂Cl ₂(50ml), use 10% KHSO ₃(50ml) Na is used in washing then ₂SO ₄Drying is filtered, and evaporation.Gained residue silica gel chromatography purification obtains 4-8, is flint glass shape thing. ¹H NMR (CDCl ₃) δ 8.31 (t, J=6Hz, 1H); 7.62 (d, J=8.5Hz, 1H); 7.01 (s, 1H); 4.43 (t, J=6.6Hz, 2H); 4.11 (m, 1H); 3.92 (d, J=12Hz, 2H); 3.83 (s, 3H); 3.80 (t, J=6.6Hz, 2H); 3.51 (t, J=7.3Hz, 2H); 3.65 (m, 2H); 3.51 (t, J=6.8Hz, 2H); 2.96 (t, J=7.2Hz, 2H); 1.70 (d, J=11Hz, 2H); 1.53 (m, 2H); 1.60-1.49 (eclipsed m, 5H); 1.40 (s, 9H); 1.28 (q, J=7.1Hz, 2H); 1.05 (m, 2H); 0.79 (t, J=7.1Hz, 3H).

Flow process 5

Flow process 5 (continuing)

1-(3-bromopropyl) pyrazoles-3,5-dioctyl phthalate dimethyl ester (5-1)

By to the described method of 3-2, use 1, the 3-dibromopropane makes chemical compound 5-1, is white crystalline solid. ¹H?NMR(CDCl ₃)δ7.38(s，1H)；4.95(t，J＝8.2Hz，2H)；3.95(s，3H)；3.92(s，3H)；3.75(t，J＝8.5Hz，2H)2.51(m，2H).

1-(3-azido propyl group) pyrazoles-3,5-dioctyl phthalate dimethyl ester (5-2)

Use NaN ₃(0.883g 13.8mmol) handles 5-1 (1.0g, 10ml DMSO solution 3.45mmol), and mixture stirred 5 hours at 25 ℃.Then, reactant mixture 100ml H ₂Ethyl acetate (3 * 100ml) extractions are used in the O dilution then.The organic extract liquid water that merges (2 * 100ml) and saline (1 * 100ml) washs, and uses Na ₂SO ₄Drying, evaporation obtains the 5-2 of colorless oil.

5,6,7,8-tetrahydrochysene-4-oxo-4H-pyrazolo [1,5-a] [1,4] diaza -2-yl]-methyl formate (5-3)

5-2 (851mg, the anhydrous EtOH solution of 100ml 3.25mmol) is handled with 100mg 10%Pd/C, and with this mixture in jolting 5 hours in the Pa Er hydrogenator under the 45psi condition.Catalyst is removed through diatomite filtration, and evaporated filtrate obtains the 800mg colorless oil.This material is analyzed through NMR, shows that it is 1-(3-aminopropyl) dimethyl pyrazole-3, the mixture of 5-dicarboxylic acid esters and ring-type diaza ketone (diazapineone).This mixture is dissolved in the 50ml benzene, refluxed 15 hours, then evaporation.The gained brown solid is through CH ₂Cl ₂/ hexane recrystallization obtains white solid 5-3.m.p.＝220-221℃. ¹H?NMR(CDCl ₃)δ7.36(s，1H)；6.42(brt，1H)；4.58(t，J＝8.0Hz，2H)；3.95(s，3H)；3.39(dt，J＝7.2Hz，2H)；2.31(m，2H).

5,6,7,8-tetrahydrochysene-4-oxo-5-[2-(N-Boc-piperidin-4-yl) ethyl]-4 H-pyrazolos [1,5-a] [1,4] diaza -2-yl]-methyl formate (5-5)

To 5-3 (175mg, add in 50ml DMF solution 0.83mmol) 60%NaH (36mg, 0.91mmol), with mixture at N ₂Stirred 30 minutes at-15 ℃ down.In this mixture, in 20 minutes, be added dropwise to 2-(N-Boc-piperidin-4-yl) ethyl iodide (5-4) (283mg, 25ml DMF solution 0.83mmol).Gained solution stirred 30 minutes at-15 ℃, was warmed to room temperature then, and stirred and spend the night.Reduction vaporization falls DMF, and residue is dissolved in the ethyl acetate again, filters, and through the silica gel chromatography purification, makes eluant with ethyl acetate, obtains pure 5-5, is glassy mass. ¹H?NMR(CDCl ₃)δ7.24(s，1H)；4.50(t，J＝7.0Hz，2H)；3.93(brd，J＝12Hz，2H)；3.94(s，3H)；3.61(t，J＝5.3Hz，2H)；3.42(t，J＝7.3Hz，2H)；2.7(brt，J＝6.3Hz，2H)；2.3(m，2H)；1.55(d，J＝12.5Hz，2H)；1.38(m，2H)；1.33-1.25(m，1H)；1.27(s，9H)；1.01(m，2H).

5,6,7,8-tetrahydrochysene-4-oxo-5-[2-(N-Boc-piperidin-4-yl) ethyl-4H-pyrazolo [1,5-a] [1,4] diaza -2-yl]-formic acid (5-6)

5-5 (166mg, 10ml CH 0.395mmol) ₃(0.435mg 0.43mmol) handles OH solution with 1N NaOH.Gained solution at room temperature stirred 18 hours, and CH is removed in decompression then ₃OH, remaining water is used CH with 10% aqueous citric acid solution acidify ₂Cl ₂(2 * 50ml) extractions.The organic extract liquid Na that merges ₂SO ₄Drying concentrates then, obtains white solid 5-6. ¹H?NMR(CDCl ₃)δ7.29(s，1H)；4.52(t，J＝7.0Hz，2H)；4.12(brd，J＝12Hz，2H)；3.94(s，3H)；3.61(t，J＝5.3Hz，2H)；3.42(t，J＝7.3Hz，2H)；2.7(brt，J＝6.3Hz，2H)；2.3(m，2H)；1.55(d，J＝12.5Hz，2H)；1.38(m，2H)；1.33-1.25(m，1H)；1.27(s，9H)；1.01(m，2H).

2 (S)-[(normal-butyl sulfonyl) amino]-3-[[[5,6,7,8-tetrahydrochysene-4-oxo-5-[2-(N-Boc-piperidin-4-yl) ethyl]-4H-pyrazolo [1,5-a] [1,4]-diaza -2-yl] carbonyl] alanine ethyl ester (5-7)

5-6 (147mg, 0.36mmol) and CH ₂Cl ₂The mixture of/5ml with 2 (S)-positive fourth sulfonamido-3-amino-ethyl propionate (A-5) (115mg, 0.40mmol), HOBT (49mg, 0.36mmol) and Et ₃N (0.10ml, 0.724mmol) and 50mlCH ₂Cl ₂Mixture process, this solution is at N ₂Under the atmosphere stirring at room 18 hours.The saturated NaHCO of reaction solution ₃, H ₂O, 10% citric acid, H ₂(Na is used in each 1 * 20ml) washing successively for O and saline ₂SO ₄Drying is filtered and evaporation.Gained fining glass shape thing uses 5%CH through the silica gel chromatography purification ₃OH/EtOAc makes eluant, obtains pure 5-7. ¹H NMR (CDCl ₃) δ 7.32 (s, 1H); 7.24 (t, J=6.8Hz, 1H); 5.54 (t, J=7.2Hz, 1H; 4.43 (t J=7.8Hz, 2H); 4.32 (m, 1H); 4.28 (q, J=7.1Hz, 2H); 4.10 (brd, J=12Hz, 2H); 3.85 (m, 2H); 3.61 (t, J=5.3Hz, 2H); 3.42 (t, J=7.3Hz, 2H); 3.03 (t, J=7.1Hz, 2H); 2.7 (brt, J=6.3Hz, 2H); 2.3 (m, 2H); 1.65-1.45 (eclipsed m, 7H); 1.38 (m, 2H); 1.33-1.25 (m, 1H); 1.37 (s, 9H); 1.30 (t, J=7.4Hz, 3H); 1.01 (m, 2H); 0.96 (t, J=7.3Hz, 3H).

2 (S)-[(normal-butyl sulfonyl) amino]-3-[[[5,6,7,8-tetrahydrochysene-4-oxo-5-[2-(piperidin-4-yl) ethyl]-4H-pyrazolo [1,5-a] [1,4]-diaza -2-yl] carbonyl] alanine (5-8)

To 5-7 (100mg, 10ml CH 0.156mmol) ₃Add in the OH solution 1NNaOH (160ml, 0.16mmol) and H ₂O (10ml).Gained solution was stirring at room 3.5 hours, and CH is removed in decompression then ₃OH.Remaining water is used CH with 10% aqueous citric acid solution acidify ₂C1 ₂(2 * 50ml) extractions.The organic extract liquid Na that merges ₂SO ₄Drying, evaporation obtains required acid then. ¹H NMR (CDCl ₃) δ 17.24 (t, J=6.8Hz, 1H); 7.28 (s, 1H); 6.0 (d, J=7.2Hz, 1H); 4.43 (t, J=7.8Hz, 2H); 4.32 (m, 1H); 4.10 (brd, J=12Hz, 2H); 3.85 (m, 2H); 3.61 (t, J=5.3Hz, 2H); 3.42 (t, J=7.3Hz, 2H); 3.03 (t, J=7.1Hz, 2H); 2.7 (brt, J=6.3Hz, 2H); 2.3 (m, 2H); 1.65-1.45 (eclipsed m, 7H); 1.38 (m, 2H); 1.33-1.25 (m, 1H); 1.37 (s, 9H); 1.01 (m, 2H); 0.96 (t, J=7.3Hz, 3H).

Should acid (89mg) in the 15ml ethyl acetate, be cooled to 0 ℃, and bubbling feeding HCl gas reaches 3 minutes.Reactant mixture is warmed to room temperature, stirred 30 minutes, on Rotary Evaporators, be evaporated to dried then.Remaining white solid is developed with ether, filters, and vacuum drying (is used P ₂O ₅), obtain 5-8, be white solid. ¹H NMR (DMSO-d ₆) δ 8.95 (brs, 1H); 8.33 (t, J=5.7Hz, 1H); 7.64 (d, J=9Hz, 1H); 7.02 (s, 1H); 4.35 (t, J=5.1Hz, 2H); 4.10 (m, 1H); 3.81 (t, J=5.2Hz, 2H); 3.6-3.4 (m, 4H); 3.21 (d, J=10.5Hz, 2H); 2.95 (t, J=7.8Hz, 2H); 2.81 (brm, 2H); 1.96 (d, J=11Hz, 2H); 1.62-1.2 (eclipsed multiplet, 9H); 0.80 (t, J=7.3Hz, 2H).

Flow process 6

6-1A, cis

6-1B, trans

3-oxo-2-[2-(N-Boc-piperidin-4-yl) ethyl] octahydro imidazo [1,5-a] pyridine-6-yl] Ethyl formate (6-1A, 6-1B)

(514mg 1.1mmol) is dissolved in the 25ml acetone, adds 10mlH with 2-6 ₂O, and with mixture heated to 60 ℃, under this temperature, kept 3.5 hours.Acetone is removed in decompression, filters the yellow mercury oxide of gained.This thick material is dissolved in the 100ml toluene, and refluxes 3 hours, and evaporation toluene gets yellow solid.

With this solid (500mg 1.11mmol) is dissolved in the 100ml ethanol, adds 75mg10%Pd/C, in the Pa Er hydrogenator under the 55psi condition with mixture jolting 13 hours.Remove catalyst with diatomite filtration, evaporating solvent.The gained colorless oil is carried out the silica gel chromatography purification, with 70% ethyl acetate/30% hexane eluting, obtains the trans product 6-1B of 268mg cis reduzate 6-1A and 132mg.Isomer 6-1A ¹H NMR (CDCl ₃) δ 4.15 (m, 1H); 4.06 (m, 2H); 3.68 (s, 3H); 3.41 (m, 2H); 3.22 (m, 1H); 2.90 (m, 1H); 2.80 (m, 2H); 2.68 (m, 2H); 2.43 (m, 1H); 2.16 (m, 1H); 1.81 (m, 1H); 1.69 (m, 2H); 1.60 (m, 1H); 1.45 (s, 9H); 1.43 (m, 3H); 1.39 (m, 1H); 1.11 (m, 2H). isomer 6-1B ¹H NMR (CDCl ₃) δ 4.34 (m, 1H); 4.06 (m, 2H); 3.69 (s, 3H); 3.40 (m, 3H); 3.36 (m, 1H); 3.30 (m, 1H); 3.11 (m, 1H); 2.89 (m, 1H); 2.84 (m, 1H); 2.67 (m, 2H); 2.63 (m, 1H); 2.30 (m, 1H); 1.69 (m, 2H); 1.67 (m, 2H); 1.60 (m, 1H); 1.45 (s, 9H); 1.41 (m, 1H); 1.09 (m, 2H).

(±)-cis-[[3-oxo-2-[2-(N-Boc-piperidin-4-yl) ethyl] octahydro imidazo [1,5-a] is given a tongue-lashing pyridine-6-yl] carbonyl] amino] the propanoic acid tert-butyl ester (6-2)

By to the described method of 1-6, use ¹N NaOH and CH ₃OH/H ₂The mixture hydrolysis 6-1A of O obtains required acid.This acid obtains 6-2 by to described method of 2-8 and the coupling of the Beta-alanine tert-butyl ester. ¹H?NMR(CDCl ₃)δ6.32(t，1H)；4.06(m，2H)；3.98(m，1H)；3.56(m，5H)；3.21(m，2H)；2.91(m，1H)；2.92(m，1H)；2.80(m，2H)；2.40(t，2H)；2.23(m，1H)；2.09(m，1H)；1.81(m，3H)；1.69(m.2H)；1.60(m，1H)；1.45(s，18H)；1.43(m，3H)；1.11(m，2H).

(±)-cis-3-oxo-2-[2-(piperidin-4-yl) ethyl] octahydro imidazo [1,5-a] pyridine-6-yl] carbonyl] alanine (6-3)

(65.3mg 0.13mmol) is dissolved in the anhydrous CH of 10ml with 6-2 ₂Cl ₂In, and be cooled to 0 ℃.Add trifluoroacetic acid (0.200ml), with solution stirring 1 hour, reduction vaporization obtained pure 6-3 then. ¹H?NMR(CD ₃OD)δ4.06(m，2H)；3.91(m，1H)；3.56(m，5H)；3.21(m，2H)；2.91(m，1H)；2.92(m，1H)；2.80(m，2H)；2.40(t，2H)；2.23(m，1H)；2.09(m，1H)；1.81(m，3H)；1.68(m，2H)；1.60(m，1H)；1.28(m，3H)；0.91(m，2H).

Flow process 7

2 (S)-[(N-CBZ-amino-sulfonyl) amino]-3-[[[[4,5,6,7-tetrahydrochysene-4-oxo-5-[2-(N-BOC-piperidin-4-yl) ethyl] pyrazolo [1,5-a] pyrazine-2-yl] carbonyl] amino] methyl propionate (7-1)

To Carbimide. chloro sulfonamides ester (45.1 μ l, add in 0 ℃ of solution of dichloromethane 0.508mmol) benzyl alcohol (53ml, 0.508mmol).Be reflected at 0 ℃ and wore out 90 minutes, add and contain triethylamine (142ml, 4-3 1.02mmol) (250mg, dichloromethane solution 0.508mmol).Make reactant mixture be warmed to room temperature, and stir spend the night (18 hours).With the sodium bisulfate aqueous solution reactant mixture is transferred to pH=3.0, (3 * 10ml) extractions merge organic extract liquid to product, are concentrated in the enterprising circumstances in which people get things ready for a trip spectrometry of silica gel purification (eluant is 95% dichloromethane, 5% methanol), obtain 7-1 with dichloromethane. ¹H?NMR(300MHz，CDCl ₃)δ1.40(s，9H)，2.65(t，2H)，3.65(s，3H)，5.10(s，2H)，6.65(d，1H)，7.2-7.5(m，6H)，8.90(s，1H)

2 (S)-[(N-CBZ-amino-sulfonyl) amino]-3-[[[[4,5,6,7-tetrahydrochysene-4-oxo-5-[2-(N-Boc-piperidin-4-yl) ethyl] pyrazolo [1,5-a] pyrazine-2-yl] carbonyl] alanine (7-2)

In the THF of 7-1 (100mg) (5ml) solution, add 1N LiOH (0.6ml), with this mixture stirring at room 18 hours.Reactant mixture is handled by adding sodium bisulfate aqueous solution (pH=3.0), and (2 * 15ml) extract with ethyl acetate with product.Concentrated extract obtains required acid. ¹H?NMR1.4(s，9H)，2.6(br，t，2H)，7.1-7.2(br，m，5H)，7.3(s，1H).

This acid is dissolved among the EtOAc, is cooled to-5 ℃, handle with HCl (gas).Concentrated reaction mixture is used ethyl acetate rinse, obtains 7-2, mp＞200 ℃ (decomposition).

CHN analytical calculation value: C, 44.86; H, 5.92; N, 14.20

Measured value: C, 44.50; H, 6.09; N, 13.80

2 (S)-(amino-sulfonyl) amino-3-[[[4,5,6,7-tetrahydrochysene-4-oxo-5-[2-(piperidin-4-yl) ethyl] pyrazolo [1,5-a] pyrazine-2-yl] carbonyl] alanine (7-3)

Handle methanol (20ml) solution of 7-2 (70mg) with 10% Pd/C (35mg), and with this mixture hydrogenation (1 atmospheric pressure) spend the night (18 hours).This mixture with concentrated, obtains 46mg grease after filtration.This grease is dissolved in the ethyl acetate, is cooled to 0 ℃, and bubbling feeding HCl gas reached more than 30 minutes.Concentrated reaction mixture obtains 7-3.Be white solid, mp＞200 ℃, FAB MS, M+1=458.

Flow process 8

1-(2-bromoethyl) pyrroles-2,4-dicarboxylate (8-2)

With pyrroles-2, and the 4-dicarboxylate (5.50g, oxolane 29.4mmol) (200ml) solution cools off in ice bath, and (6.5g, oxolane 68.6mmol) (50ml) suspension adds continuously with NaH (60%).Reaction bulb is warmed to room temperature.After at room temperature stirring 1 hour, the adding glycol dibromide (25.2ml, 294mmol), and with mixture backflow 24 hours.In reaction flask, add entry (50ml).Evaporate this mixture with Rotary Evaporators, to remove oxolane.Add saturated sodium bicarbonate solution (100ml) in residue, (4 * 50ml) extract gained solution with dichloromethane.The organic extract liquid anhydrous sodium sulfate drying that merges.Desiccant is removed after filtration, and filtrate is evaporated through Rotary Evaporators, obtains yellow solid.This material obtains 8-2 with 80: 20 recrystallization of hexane-ethyl acetate, is yellow solid. ¹H?NMR(DMSO-d ₆)δ7.83(d，1H)；7.15(d，1H)；4.69(t，2H)；4.25(m，4H)；3.78(t，2H)；3.31(H ₂O)；1.29-1.22(m，6H).

[4,5,6,7]-tetrahydrochysene-4-oxo-5-[2-(N-Boc-piperidin-4-yl) ethyl] pyrrolo-[1,5-a] pyrazine-2-yl] Ethyl formate (8-3)

With alkyl bromide 8-2 (3.30g, 10.4mmol, 1.0 equivalents), 2-4 (3.52g, 15.5mmol, 1.5 equivalents), potassium iodide (5.18g, 31.2mmol), diisopropyl ethyl amine (5.42ml, 31.2mmol, 3.0 equivalents) and acetonitrile (50ml) mix.With this suspension reflux 24 hours, be rotated evaporation then, to remove acetonitrile.Add saturated sodium bicarbonate solution (100ml), (5 * 50ml) extract with ethyl acetate with this solution.The organic extract liquid anhydrous sodium sulfate drying that merges, the simmer down to brown oil.Crude product silica gel column chromatography purification.Then with the dichloromethane elution chromatography post that contains 1% methanol, obtain pure 8-3 with dichloromethane, be white solid. ¹H?NMR(CDCl ₃)δ7.32(d，1H)；7.29(d，1H)；4.28(q.2H)；4.19-4.00(m，4H)；3.7-3.6(t，2H)；3.6-3.5(t，2H)；2.68(t.2H)；1.8-1.7(m，br，2H，H ₂O)；1.6-1.4(s，m，11H)；1.33(t，3H)；1.2-1.1(m，2H).

[4,5,6,7]-tetrahydrochysene-4-oxo-5-[2-(N-Boc-piperidin-4-yl) ethyl] pyrrolo-[1,5-a] pyrazine-2-yl] the alanine tert-butyl ester (8-4)

On equipment, mix in the 50ml round-bottomed flask of a magnetic stirring bar 8-3 (620mg, 1.54mmol), lithium hydroxide monohydrate (160mg, 4.00mmol, 2.6 equivalents), water (15ml) and methanol (10ml).This solution was at room temperature stirred 4 hours, be heated to 90 ℃ then, and under this temperature, kept 1 hour.Any remaining methanol is removed by rotary evaporation, aqueous residue 10%K ₂SO ₄Ethyl acetate (4 * 50ml) extractions are used in acidify then.The organic extract liquid anhydrous sodium sulfate drying that merges filters, and evaporation, obtains required acid, is white solid. ¹H?NMR(DMSO-d ₆)δ7.51(d，2H)；6.84(d，1H)；4.19(m，2H)；3.90(d，br，2H)；3.63(m，2H)；3.43(m，2H)；3.4-3.2(H ₂O)；2.6-2.8(br，2H)；1.66(d，2H)；1.43(m，2H)；1.36(s，9H)；1.1-0.9(m，2H).

In being housed, mixes by the 100ml round-bottomed flask of a magnetic stirring bar above-mentioned acid (150mg, 554mmol), EDC (117mg, 0.609mmol), I-hydroxybenzotriazole (82.2mg0.609mmol), triethylamine (0.300ml, 1.11mmol, 4.0 equivalent), the Beta-alanine tert-butyl ester (111mg, 0.609mmol) and dichloromethane (10ml).Gained solution at room temperature stirred spend the night.Rotary evaporation falls solvent from reaction flask, gained residue silica gel column chromatography purification, chromatographic column with dichloromethane, contain the dichloromethane of 2% methanol, then with containing the dichloromethane eluting of 4% methanol.Merge the fraction that contains product, obtain 8-4, be white solid. ¹H?NMR(CDCl ₃)δ7.32(d，1H)；7.04(d，1H)；6.58(m，1H)；4.2-4.0(m，4H)；3.7-3.5(m，6H)；2.68(t.2H)；2.51(t，2H)；1.70(m，3H，H2O)；1.53(m，2H)；1.45(s，9H)；1.21-1.0(m，2H).

4,5,6,7-tetrahydrochysene-4-oxo-5-[2-(piperidin-4-yl) ethyl] [1,5-a] pyrazine-2-yl] carbonyl] alanine (8-5)

Mixed ester 8-4 in the 50ml round-bottomed flask (180mg, 0.347mmol) and ethyl acetate (10ml).This suspension is cooled off in ice bath.Bubbling feeding hydrogen chloride reaches 1.5 minutes in suspension.Reaction flask is warmed to room temperature, and solvent is removed through vacuum filtration then, obtains 8-5, is white solid, mp.248-249 ℃. ¹H?NMR(DMSO-d ₆)δ9.0-8.5(br，2H)；8.05(m，1H)；7.42(d，1H)；7.05(d，1H)；4.15(m，2H)；3.62(m，2H)；3.5-3.3(m，4H，H ₂O)；3.25-3.15(d，br，2H)；2.85-2.7(br，2H)；2.5-2.4(m，4H)；1.81(d，2H)；1.6-1.4(m，3H)；1.4-1.2(m，2H).

Flow process 9

2 (S)-amino-3-(N-Boc-amino) ethyl propionates (9-1)

(10g 96.2mmol) is dissolved in the dehydrated alcohol (200ml), and this solution is saturated with anhydrous HCl gas then, and reflux is 2.5 hours then with commercially available 2 (S)-3-diaminopropionic acids (Fluka).Remove and desolvate residue EtOH/Et ₂The O recrystallization obtains the ethyl ester dihydrochloride, is the white solid of moisture absorption.

(5g 24.3mmol) is suspended in the CH that is cooled to-50 ℃ with this material ₂Cl ₂(200ml).Then, (7.0ml 51mmol), and stirred this mixture 5 minutes to add triethylamine.In 30 fens clock times, the company's of being added dropwise to Bis(tert-butoxycarbonyl)oxide (5.30g, 100ml CH 24.3mmol) ₂Cl ₂Solution, and this mixture stirred 1.5 hours at-50 ℃, damp and hot then to room temperature.(Na is used in 2 * 100ml) washings to this solution with water then ₂SO ₄Drying, and evaporation.Gained residue silica gel chromatography purification (80: 20 CH ₂Cl ₂/ CH ₃OH), obtain pure 9-1. ¹H?NMR(300MHz，CDCl ₃)6.03(brt，1H)；4.35(m，1H)；3.85(m，2H)；3.23(m，2H)；1.21(t，3H).

2 (S)-normal-butyl aminosulfonyl amino-3-(N-BOC-amino) ethyl propionates (9-2)

(261ml, 3.23mmol) (406mg 2.37mmol) handles 9-1 (500mg, dichloromethane 2.15mmol) (10ml) solution with normal-butyl sulfonamides chlorine with pyridine.This solution at room temperature stirred 3 hours, inclined then to silica gel, with 30% acetone/hexane eluting, obtained pure 9-2, was white solid. ¹H NMR (300MHz, CDCl ₃) δ 5.43 (d, 1H); 4.98 (t, 1H); 4.40 (brs, 1H); 4.10 (q, 2H); 4.05 (m, 1H); 3.56 (m, 2H); 3.08 (m, 2H); 1.8-1.2 (eclipsed multiplet, 16H); 0.90 (t, 3H).

2 (S)-(normal-butyl aminosulfonyl amino)-3-alanine ethyl ester (9-3)

With 9-2 (612mg, ethyl acetate 1.65mmol) (50ml) solution is cooled to-5 ℃, bubbling feeds anhydrous HCl and reaches 30 minutes, and reactant mixture is concentrated, and filters to isolate product, 9-3, be white solid. ¹H?NMR(300MHz，DMSO-d ₆)δ5.82(d，1H)；4.56(brs，1H)；4.20(q，2H)；4.02(m，1H)；3.45(m，2H)；3.01(m，2H)；1.9-1.36(m，7H)；0.93(t，3H).

2 (S)-[(normal-butyl amino-sulfonyl) amino]-3-[[4,5,6,7-tetrahydrochysene-4-oxo-5-[2-(piperidin-4-yl) ethyl] pyrazolo [1,5-a] pyrazine-2-yl] carbonyl] alanine ethyl ester (9-4)

By to the described method of 1-6, in DMF, make 9-3 and 3-3, EDC and HOBT coupling, obtain 9-4. ¹H?NMR(300MHz，CDCl ₃)δ7.29(s，1H)；7.19(t，1H)；5.43(d，1H)；4.41(t，2H)；4.26(q，2H)；4.20(m，1H)；4.08(d，2H)；3.86(m，2H)；3.76(m，2H)；3.60(t，2H)；3.08(t，2H)；2.68(t，2H)；1.78(d，2H)；1.6-1.08(m，8H)；1.43(s，9H)；0.92(t，3H).

2 (S)-[(normal-butyl amino-sulfonyl amino]-3-[(4,5,6,7-tetrahydrochysene-4-oxo-5-(piperidin-4-yl) ethyl] pyrazolo [1,5-a] pyrazine-2-yl] carbonyl] alanine (9-5)

By to the described method of 5-7, with NaOMe hydrolysis 9-4, separate thick acid, use the EtOAc solution-treated of HCl subsequently, obtain 9-5, be white solid, mp155-160 ℃.

Flow process 10

N-[2-(5-methoxycarbonyl group) pyridylmethyl]-the Beta-alanine tert-butyl ester (10-1)

With 2-3 (871mg, 3.79mmol), Beta-alanine tert-butyl ester HCl (2.7g, 15mmol) and K ₂CO ₃(4.5g is 30mmol) at the anhydrous CH of 100ml ₃Mixture among the CN places the 250ml flask, and refluxes 3 hours, cools off then and filters.Concentrating under reduced pressure filtrate is carried out the silica gel chromatography purification, makes eluant with EtOAc, obtains 10-1, is flint glass shape thing. ¹H NMR (CDCl ₃) δ 9.18 (d, J=1.4Hz, 1H); 8.1 (dd, J=1.4 and 6.8Hz, 1H); 7.39 (d, J=6.8Hz, 1H); 4.08 (s, 2H); 3.95 (s, 3H); 3.04 (t.2H); 2.60 (t, 2H); 1.4 (s, 9H).

The tert-butyl group-2-(2-carboxyethyl)-1-is chloroformyl-3-oxo-2, and the 3-dihydro-imidazol-is [1,5-a] pyridine-6-formic acid esters (10-2) also

(800mg 2.71mmol) is dissolved in the 50ml toluene, adds N, and (2.0ml 16.7mmol), is cooled to 0 ℃ with solution to accelerine with 10-1.To wherein, in 30 minutes, be added dropwise to triphosgene (1.7g, 15ml toluene solution 5.7mmol).Then, this solution is warmed to 25 ℃, and stirred 3.0 hours, use 1N HCl, water and saline (each 50ml) washed twice afterwards, use Na ₂SO ₄Drying, and evaporation obtain 10-2, are yellow crystalline solid. ¹H?NMR(CDCl ₃)δ8.83(d，J＝1.4Hz，1H)；8.25(d，J＝6.8Hz，1H)；7.82(dd，J＝1.4?and?6.8Hz，1H)；4.43(t，J＝7.2Hz，2H)；3.98(s，3H)；2.75(t，J＝7.2Hz，2H)；1.4(s，9H).

The tert-butyl group-2-(2-carboxyethyl)-1-cyano group-3-oxo-2,3-glyoxalidine be [1,5-a] pyridine-6-formic acid esters (10-3) also

(250mg 0.65mmol) is dissolved in 100ml CH with 10-2 ₂Cl ₂In, add 10ml ammonium hydroxide, and this two-phase mixture was stirred 1 hour, separate organic layer then, with 10% citric acid, use saline (50ml) to wash then, use Na ₂SO ₄Drying, and evaporation.

(150mg 0.42mmol) is dissolved in 100ml CH with this residue ₂Cl ₂In, during 2 hours in, divide add for three times 355.7mg hydroxide methoxycarbonyl group sulfamoyl-triethyl ammonium inner salt (Burgess reagent, 1.48mmol).With gained solution restir 1 hour at room temperature, concentrate then, at the enterprising circumstances in which people get things ready for a trip spectrometry of silica gel purification, use 1: 1 hexane/ethyl acetate to make eluant, obtain described nitrile with quantitative yield.This material is carried out saponification with 1N LiOH, obtain required carboxylic acid 10-3. ¹H?NMR(CDCl ₃)δ8.75(s，1H)；7.43(d，1H)；7.18(d，1H)；4.21(t，2H)；2.85(t，3H)；1.4(s，9H).

3-[1-cyano group-3-oxo-6-2-(piperidin-4-yl) ethylamino formoxyl)-2, the 3-dihydro-imidazol-is [1,5-a] pyridine-2-yl also] propanoic acid (10-4)

(170mg 0.51mmol) is dissolved in 10ml CH with 10-3 ₂Cl ₂In, add Et ₃N (71ml, 0.51mmol) and HOBT (69.3mg, 0.51mmol), EDC (98.6mg, 0.52mmol) and 2-4 (117.2mg, 0.51mmol).With this mixture at N ₂Stirred 18 hours under the atmosphere, use 10% citric acid, H then ₂Na is used in O and saline (each 10ml) washing ₂SO ₄Drying concentrates the circumstances in which people get things ready for a trip spectrometry purification of going forward side by side, obtain yellow solid (215mg, 0.43mmol).This material is through using at CH ₂Cl ₂In trifluoroacetic acid go the protection, obtain 10-4TFA salt, be yellow solid, m.p.=173 ℃. ¹H?NMR(300MHz，DMSO?d ₆)δ1.75(s，1H)；7.43(t，1H)；7.31(d，1H)；7.15(d，1H)；4.23(t，2H)；3.41(t，2H)；3.23(d，2H)；2.83(m，2H)；1.85(d，2H)；1.53(m，2H)；1.41(m，1H)；1.32(m，2H).

Flow process 11

[4,5,6,7-tetrahydrochysene-4-oxo-5-[3-(tert-butyl group propiono)] pyrazolo [1,5-a] pyrazine-2-yl] methyl formate (11-1)

With 3-2 (1.4g, 4.8mmol), Beta-alanine tert-butyl ester HCl (0.90g, 5mmol) and potassium carbonate (0.78g is 5.28mmol) at 150ml CH ₃Among the CN, at N ₂Refluxed 4.5 hours under the atmosphere, cooling is then filtered, and reduction vaporization.The gained yellow residue is carried out the silica gel chromatography purification, use 2%CH ₃OH/CH ₂Cl ₂Eluting obtains diester 11-1, is flint glass shape thing. ¹H?NMR(CDCl ₃)δ7.31(s，1H)；4.48(t，2H)；3.93(s，3H)；3.61(t，2H)；2.71(t，2H)；2.35(t，2H)；1.23(s，9H).

3-[4,5,6,7-tetrahydrochysene-4-oxo-2-(2-(piperidin-4-yl) ethylamino formoxyl] pyrazolo [1,5-a] pyrazine-5-yl] propanoic acid (11-2)

To contain LiOH (145mg, 10ml H 3.41mmol) ₂O solution joins ester 11-1 (1.0g, 10ml CH 3.1mmol) ₃In the OH solution, and, under this temperature, kept 2.5 hours cooling, and removal of solvent under reduced pressure then with this mixture heated to 60 ℃.Remaining residue is used CH with 10% citric acid acidify ₂Cl ₂(2 * 100ml) extractions.The organic extract liquid H that merges ₂The O washing, dry and evaporation obtains required acid, is flint glass shape thing. ¹H?NMR(CDCl ₃)δ7.21(s，1H)；4.48(t，2H)；3.63(t，2H)；2.71(t，2H)；2.32(t，2H)；1.23(s，9H).

Should acid (500mg 1.62mmol) is dissolved in 10ml CH ₂Cl ₂In.Add HOBt (220mg, 1.62mmol) and EDC (309mg, 1.62mmol) and 2-4 (356mg, 1.63mmol).With this mixture at N ₂Stirred 16 hours under the atmosphere, use 10% citric acid, H then ₂Na is used in O and saline (each 10ml) washing ₂SO ₄Drying concentrates the circumstances in which people get things ready for a trip spectrometry purification of going forward side by side, and obtains the colourless foam body.This material goes protection with HCl in ethyl acetate, obtain the HCl salt of 11-2, is white solid, mp=192-194 ℃. ¹H NMR (300MHz, DMSO d ₆) δ 9.0 (br, s, 1H); 8.35 (m, 1H); 6.99 (s, 1H); 4.38 (t, J=6.0Hz, 2H); 3.83 (t, J=5.5Ht, 2H); 3.64 (t, J=7.1Hz, 2H); 3.30-3.1 (m, 4H); 2.85-2.65 (q, 2H); 2.54 (t, J=7.1Hz, 2H); 1.85-1.75 (s, br, 2H); 1.60-1.20 (eclipsed m, 5H).

Flow process 12

2-(4-pyridine radicals) ethylamine (12-1)

With NH ₄The 200ml H of Cl ₂O solution places the 1L flask.(56.4ml is 0.52mol) with 150ml CH to add 4-vinylpridine ₃OH, and to this mixture 60 ℃ the heating 18 hours.In ice bath, reaction solution is cooled to 0 ℃, adds 30% NaOH and make it to be alkalescence.This alkaline solution CH ₂Cl ₂(5 * 100ml) extractions, the dry extract that merges, evaporation afterwards.This residue of vacuum distilling obtains 12-1, is colourless liquid. ¹H?NMR(300MHz，CDCl ₃)δ8.53(d，3＝6.1Hz，2H)；7.25(d.J＝6.1H2，2H)；3.02(t，2H)；2.77(t，2H)；1.4(brs，2H).

4,5,6,7-tetrahydrochysene-4-oxo-5-[2-(pyridin-4-yl) ethyl] pyrazolo [1,5-a] pyrazine-2-yl] methyl formate (12-2)

With 3-2 (1.4g, 4.8mmol), 4-(2-amino-ethyl pyridine) (0.645g, 5.28mmol) and potassium carbonate (0.78g, 150ml CH 5.28mmol) ₃CN solution is at N ₂Refluxed 4.5 hours under the atmosphere, cooling is then filtered and reduction vaporization.The gained yellow residue is dissolved among the 50ml DMF again, handles, and 90 ℃ of heating 3 hours, concentrating under reduced pressure at the enterprising circumstances in which people get things ready for a trip spectrometry of silica gel purification, used 20% CH then with NaH (200mg, 60% oil suspension) ₃OH/CH ₂Cl ₂Eluting obtains ester 12-2, is light yellow glassy mass (0.91g, 3.0mmol, 68%). ¹H?NMR(CDCl ₃)δ8.32(d，2H)；7.52(d，2H)；7.34(s，1H)；4.48(t，2H)；3.91(s，3H)；3.61(t，2H)；2.71(t，2H)2.35(t，2H).

2 (S)-[(p-toluenesulfonyl) amino]-3-[[[4,5,6,7-tetrahydrochysene-4-oxo-5-[2-(4-pyridine radicals) ethyl] pyrazolo [1,5-a] pyrazine-2-yl] t-butyl formate (12-3)

To contain LiOH (130mg, 10ml H 3.05mmol) ₂O solution joins 12-2 (910mg, 10ml CH 3.0mmol) ₃In the OH solution,, under this temperature, kept 2.5 hours this mixture heated to 60 ℃, cooling then, removal of solvent under reduced pressure, remaining residue is through carrying out the ion exchange chromatography purification on the Dowex-50W resin, obtain required acid, be pale solid, mp.187 ℃.

Should acid (300mg 0.78mmol) is suspended in the 50ml dry DMF.Add A-8 (293mg, 81mmol), EDC (150mg, 0.78mmol), HOBt (105mg, 0.78mmol) and N-methylmorpholine (87ml, 0.78mmol), and with the settled solution of gained 25 ℃ of stirrings 19 hours.This solution is used saturated NaHCO successively with 100ml EtOAc dilution ₃, H ₂Na is used in O and saline (25ml) washing ₂SO ₄Drying, evaporation obtains 12-3. ¹H?NMR(300MHz，DMSO-d ₆)δ8.63(d，2H)；7.80(d，2H)；7.58(d，2H)；7.29(t，1H)；6.93(s，1H)；5.95(d，2H)；4.40(t，2H)；4.08(m，1H)；3.86-3.74(m，4H)；3.35-3.20(m，2H)；3.10(t，2H)；1.30(s，9H)；

2 (S)-[(p-toluenesulfonyl) amino]-3-[[[4,5,6,7-tetrahydrochysene-4-oxo-5-[2-(4-pyridine radicals) ethyl] pyrazolo [1,5-a] pyrazine-2-yl] formic acid tfa salt (12-4)

12-3 uses TFA at CH ₂Cl ₂In go the protection, use the reverse-phase chromatography purification, obtain the tfa salt of 12-4, mp.182-185 ℃. ¹H?NMR(300MHz，DMSO-d ₆)δ8.72(brd，2H)；8.19(t，1H)；8.00(d，1H)；7.81(d，2H)；7.58(d，2H)；7.19(d，2H)；6.85(s，1H)；4.40(t，2H)；4.01(m，1H)；3.83-3.76(m，4H)；3.48(m，1H)；3.26(m，1H)；3.10(t，2H).

Flow process 13

Flow process 13 (continuing)

5,6,7,8-tetrahydrochysene-4-oxo-5-[(3-cyano-phenyl) methyl-4-H-pyrazolo [1,5-a] [1,4] diaza -2-yl] methyl formate (13-1)

(3.02g, 60ml anhydrous DMF solution 14.5mmol) is cooled to 0 ℃, and (636mg 15.98mmol) handles with NaH (60% fluid) with 5-3.The gained mixture was stirred 1.5 hours at 0 ℃, drip 3-cyano-benzyl bromide (3.11g, 50ml DMF solution 15.89mmol) then.The gained mixture was stirred 18 hours at 25 ℃, with 200ml EtOAc dilution, use H then ₂O (3 * 100ml) and saline (100ml) washing.Organic layer Na ₂SO ₄Drying is filtered and evaporation.Gained solid CH ₂Cl ₂/ CH ₃The OH recrystallization obtains 13-1, is white solid. ¹H?NMR(CDCl ₃)δ7.85(s，1H)；7.78(d，J＝8Hz，1H)；7.59(d，J＝8Hz，1H)；7.46(m，1H)；7.30(s，1H)；4.80(s，1H)；4.43(t，J＝8Hz，2H)；3.89(s，3H)；3.38(t，J＝8Hz，2H)；2.09(m，2H).

2 (S)-[(p-toluenesulfonyl) amino]-3-[[[5,6,7,8-tetrahydrochysene-4-oxo-5-[(3-cyano-phenyl) methyl]-4H-pyrazolo [1,5-a] [1,4] diaza -2-yl] carbonyl] amino] the propanoic acid tert-butyl ester (13-2)

(1.5g, (5.1ml is 5.1mmol) with 100ml H with 1N LiOH for 100ml THF solution 4.04mmol) for ester 13-1 ₂O handles, and stirs 1.5 hours at 25 ℃.THF is removed in decompression, the 1N HCl acidify of aqueous residue.Filter the gained precipitate, and vacuum drying, obtain required product, be white solid. ¹H?NMR(CDCl ₃)δ7.95(s，1H)；7.73(d，1H)；7.53(d，1H)；7.43(m，1H)；7.30(s，1H)；4.85(s，2H)；4.43(t，2H)；3.31(t，2H)；2.08(m，2H).

With above-mentioned acid (1.0g, 3.23mmol) with A-9 (1.24g, 3.54mmol), HOBt (480mg, 3.54mmol), EDC (641mg, 100mg CH 3.54mmol) ₂Cl ₂Solution mixes, and (403 μ l, 3.83mmol), gained solution at room temperature stirred 16 hours, used saturated NaHCO then successively to add N-methylmorpholine ₃, 10%KHSO ₃And saline (each 100ml) washing, use Na then ₂SO ₄Drying is filtered and evaporation, and residue obtains 13-2 through silica gel chromatography purification (using the EtOAc eluting), is white solid. ¹H?NMR(300MHz，CDCl ₃)δ7.73(d，J＝6.8Hz，2H)；7.68(s，1H)；7.65(d，1H)；7.51(m，1H)；7.37(d，J＝6.8Hz，2H)；7.20(d，1H)；7.18(t，1H)；5.63(d，J＝6.5Hz，1H)；4.80(s，2H)；4.78(m，1H)：4.45(t，2H)；3.85(m，1H)；3.08(m，1H)；3.40(t，2H)；2.43(s，3H)；2.19(m.2H)；1.65(s，9H).

2 (S)-[(p-toluenesulfonyl) amino]-3-[[[5,6,7,8-tetrahydrochysene-4-oxo-5-[(3-cyano-phenyl) methyl]-4H-pyrazolo [1,5-a] [1,4] diaza -2-yl] carbonyl] amino] propionic ester (13-3)

The CH of 13-2 ₂Cl ₂(15ml) solution is handled with 5ml TFA.This solution was stirred 2.5 hours at 0 ℃, and evaporation obtains 13-3 then, is colorless solid. ¹H?NMR(300MHz，CDCl ₃)δ7.73(d，J＝6.8Hz，12H)；7.65(s，1H)；7.65(d，1H)；7.50(m，1H)；7.31(d，J＝6.8Hz，2H)；7.3(d，1H)；7.28(t，1H)；6.15(d，J＝6.5Hz，1H)；4.80(s，2H)；4.63(m，1H)；4.43(t，2H)；3.82(m，1H)；3.65(m，1H)；3.48(m，2H)；2.43(s，3H)；2.19(m，2H).

2 (S)-[(p-toluenesulfonyl) amino]-3-[[[5,6,7,8-tetrahydrochysene-4-oxo-5-[(3-amidino groups phenyl) methyl]-4H-pyrazolo [1,5-a] [1,4] diaza -2-yl] carbonyl] amino] propanoic acid (13-4)

(400mg 0.73mmol) is dissolved in 10ml pyridine and Et with 13-3 ₃In 4: 1 mixture of N.This solution SH ₂Saturated, and stir until no longer can detect nitrile (2.5 hours) by HPLC.Excessive SH ₂By nitrogen current is removed by solution.Evaporate last solution then, residue is developed with 1N HCl, and filters, and obtains yellow solid.This material is dissolved in the 15ml acetone, uses CH ₃I (250 μ l) handles, and is heated to 50 ℃ then, up to not detecting thioamides (2 hours) again by HPLC.Solvent evaporated and excessive CH ₃I is dissolved in residue again and contains (NH ₄) ₂CO ₃(144mg, CH 1.14mmol) ₃Among the OH.This solution 50 ℃ of heating 12.5 hours, is evaporated then, and 13-4 separates by preparation property reverse-phase chromatography. ¹H?NMR(300MHz，DMSO-d ₆)δ9.38(s，2H)；9.17(s，2H)；8.19(t，1H)；8.16(d，2H)；7.78(s，1H)；7.75(m，2H)；7.63(m，1H)；7.60(d，2H)；7.21(d，2H)；6.93(s，1H)；4.81(s，2H)；4.40(t，2H)；3.93(m，1H)；3.40(m，2H)；3.35(m，2H)；2.23(s，3H)；2.13(m，2H).

Flow process 14

Flow process 14 (continuing)

5,6,7,8-tetrahydrochysene-4-oxo-5-(3-chloropropyl)-4H-pyrazolo [1,5-a] [1,4] diaza -2-yl] methyl formate (14-1)

Press the described method of 13-1, (1.5ml, 10.5mmol) (2.0g 9.5mmol), obtains 14-1 to alkylation 5-3, is white solid with 1-chloro-3-N-Propyl Bromide. ¹H?NMR(300MHz，CDCl ₃)δ7.28(s，1H)；4.58(t，2H)；3.93(s，3H)；2.78(t，2H)；2.68(t，2H)；2.46(t，2H)；3.27(m，2H)；2.18(m，2H).

5,6,7,8-tetrahydrochysene-4-oxo-5-(3-azido propyl group)-4H-pyrazolo [1,5-a] [1,4] diaza -2-yl] formic acid (14-2)

With above-mentioned chloride (909mg, 3.2mmol) and NaN ₃(620mg, DMF 9.5mmol) (15ml) solution at room temperature stirred 36 hours.This solution is used H with ethyl acetate (50ml) dilution ₂(Na is used in 3 * 50ml) washings to O then ₂SO ₄Drying is filtered, and evaporation, obtains azide, is white solid.By common this material of mode hydrolysis, obtain 14-2, be white solid. ¹H?NMR(300MHz，DMSO-d ₆)δ7.00(s，1H)；4.41(t，2H)；3.52(t，2H)；3.41(t，2H)；3.26(t，2H)；2.20(m，2H)；1.80(m，2H).

2 (S)-(tolysulfonyl amino)-3-[5,6,7,8-tetrahydrochysene-4-oxo-5-(3-aminopropyl)-4H-pyrazolo [1,5-a] [1,4] diaza -2-yl] carbonyl] amino] the propanoic acid tert-butyl ester (14-3)

By to the described method of 13-2, make sour 14-2 and A-9 coupling, obtain required product, be white solid.This material is dissolved in the ethanol, follows with 10% Pd/C at H ₂Hydrogenation under the atmosphere obtains 14-3, is white solid. ¹H?NMR(300MHz，DMSO-d ₆)8.18(t，1H)；7.68(d，2H)；7.23(d，2H)；6.98(s，1H)；4.4(m，3H)；3.93(t，2H)；3.48-3.2(m，6H)；2.78(t，2H)；2.43(s，3H)；2.69(m，2H)；1.86(m，2H)；1.08(s，9H).

2 (S)-(tolysulfonyl amino)-3-[5,6,7,8-tetrahydrochysene-4-oxo-5-(3-guanidine radicals propyl group)-4H-gives a tongue-lashing also [1,5-a] [1,4] diaza -2-yl of azoles] carbonyl] amino] propanoic acid (14-4)

14-3 (60mg, DHF 0.1mmol) (5ml) solution with DIPEA (90 μ l, 0.5ml) and 3-1-carbonamidine (30mg 0.5mmol) handles and heated 12 hours at 80 ℃.Evaporating solvent, residue neutral alumina chromatography purification (CH ₂Cl ₂/ CH ₃OH/NH ₄OH, 80/20/1), obtain required product, be white solid, this material goes protection in due form with TFA, with preparation property reacting phase chromatography purification, obtains (14-4), is white solid. ¹H?NMR(300MHz，D ₂O)δ8.2(t，1H)；7.58(s，2H)；7.18(d，2H)；4.38(t，2H)；3.51(m，5H)；3.45(t，2H)；3.2(m，1H)；3.18(m，2H)；2.10(s，3H)；2.08(m，2H)；1.8(m，2H).

2 (S)-(tolysulfonyl amino)-3-[5,6,7,8-tetrahydrochysene-4-oxo-5-[3-[N-(imidazoline-2-yl) amino] propyl group]-4H-pyrazolo [1,5-a] [1,4] diaza -2-yl] carbonyl] amino] propanoic acid (14-5)

Use makes solution and the 2-methyl mercapto-2-imidazoline hydriodate reaction of 14-3 in method described in the 14-4.This thick material goes protection with TFA, and separates 14-5 by preparation property reverse-phase chromatography. ¹H NMR (300MHz, DMSO-d ₆) 8.20 (t, 1H); 8.10 (d, 2H); 7.60 (d, 2H); 7.21 (d, 3H); 6.86 (s, 1H); 4.46 (t, 2H); 4.01 (m, 2H); 3.8-3.5 (eclipsed m, 8H); 2.23 (s, 3H); 2.10 (t, 2H); 1.80 (t, 2H).

Flow process 15

2 (S)-[(p-toluenesulfonyl) amino]-3-[[[5,6,7,8-tetrahydrochysene-4-oxo-5-[2-(N-BOC-piperidin-4-yl) ethyl]-4H-pyrazolo [1,5-a] [1,4] diaza -2-yl] carbonyl] amino] propanoic acid (15-1)

With 5-6 (5.0g, THF 12.3mmol) (150ml) solution is cooled to 0-10 ℃, with syringe add N-methylmorpholine (2.11ml, 19.2mmol).Mix after 20 minutes, (2.38ml 18.2mmol), after 0.5 hour, obtains required mixed anhydride with the gained solution stirring to drip the chloro-carbonic acid tert-butyl ester with syringe.

Have in the round-bottomed flask of magnetic stirring bar at 500ml, mix A-7 (7.00g, 27.1mmol), THF (125ml) and diisopropyl ethyl amine (4.71ml, 27.1mmol).Add low amounts of water, until forming settled solution.Gained solution is cooled off in ice bath, under vigorous stirring, this mixed anhydride suspension of disposable adding in solution.After 20 minutes, the solution that concentrates this stirring is to remove THF.Last hydrous matter filters the gained precipitate with 10% potassium acid sulfate acidify, obtains white solid.

(10 * 20cm) carry out the flash column chromatography purification for EM Science, 220-400 order with silica gel with this material, chromatographic column dichloromethane: methanol: ammonium hydroxide 98: 2: 0.2,95: 5: 0.5,90: 10: 1, use 85: 15: 1.5 eluting then, obtain pure 15-1, be white solid. ¹H?NMR(DMSO-d ₆)δ8.23(q，J＝3.40Hz，1H)；7.64(d，J＝8.20Hz，2H)；7.32(d，J＝8.20Hz)；7.2-7.0(br，1H)；6.86(s，1H)；4.36(t，J＝6.70Hz，2H)；3.89(d，br，J＝12.21Hz，2H)；3.59(m，1H)；3.47(t，J＝7.08Hz，2H)；3.5-3.1(m，br，5H，H ₂O)；2.8-2.6(br，2H)；2.33(s，3H)；2.17(t，J＝6.47Hz，2H)；1.66(d，br，J＝11.97Hz.2H)；1.55-1.45(m，br，3H)；1.37(s，9H)；1.1-0.9(m，br，2H).

2 (S)-[(p-toluenesulfonyl) amino]-3-[[[5,6,7,8-tetrahydrochysene-4-oxo-5-[2-(piperidin-4-yl) ethyl]-4H-pyrazolo [1,5-a] [1,4] diaza -2-yl] carbonyl] amino] propanoic acid (15-2)

(7.42g 11.48mmol) places the 1L round-bottomed flask that magnetic stirring bar is housed with 15-1.Add dichloromethane, reactant mixture is cooled to 0-5 ℃, under agitation bubbling feeds hydrogen chloride in suspension.After about 2 minutes, solid becomes solution, forms another kind of precipitation soon.Bubbling fed gas after 5 minutes again in suspension, and reaction bulb is warmed to room temperature.After 30 minutes, the inclusions in the concentration response bottle.The gained white solid is the hydrochlorate of 15-2, with HPLC purity assay＞99%.

The hydrochlorate of this 15-2 uses Dowex 50X8-200 ion exchange resin, and (110g 4.11meq/g) carries out the ion exchange chromatography purification.This resin washs preparation by water, methanol, water, 6N hydrochloric acid, He Shui (each 500ml).At this moment, the pH of eluant is 7.In hydrochlorate water-soluble (30ml), be applied to the top of post then.This post of water eluting.The pH of eluant becomes highly acid.When the pH of eluant returns to 7, use ammonium hydroxide: acetonitrile: 50: 25: 25 (1.5L) eluting of water post.Merge the part that contains the U.V active substance, in fine vacuum, concentrate then.Gained white foam shape thing in fine vacuum dry 8 hours obtains 15-2. ¹H?NMR(DMSO-d ₆)δ9.0-8.5(br，1H)；8.19-8.16(m，1H)；7.67(d，J＝8.18Hz，2H)；7.32(d，J＝8.18Hz，2H)；6.89(s，1H)；4.38(t，J＝6.84Hz，2H)；3.75-3.65(m，br，1H)；3.46(t，br，2H)；3.5-3.1(m，br，8H，H ₂O)；2.77(t，br，J＝11.36，2H)；2.35(s，3H)；2.17(t，J＝6.47Hz，2H)；1.80(d，br，J＝12.7Hz，2H)；1.53-1.42(m，br，3H)；1.33-1.24(m，br，2H).

Use method noted earlier,, prepared the chemical compound in time tabulation 1 particularly in method described in flow process 3 and 4.

Table 1

Table 1 (continuing)

R mp (℃) salt form

Other chemical compounds according to the method preparation that is similar to the foregoing description method are shown in down in the tabulation:

Table 2

-CN H H 180-188

Table 2 (continuing)

R A B mp(℃)

Table 3

n m mp(℃)

2 0 110-115

2 1 115-120

1 1 121-123

0 2 135-141

0 1 140-145

Table 4

The spatial chemistry of amide

NH ^*R is with respect to H ₆The trans 7.38NHSO of chemical shift (PPM) H cis 6.35H ₂C ₄H ₉Cis 6.31NHSO ₂C ₄H ₉Trans 7.29 " H ₆" be meant the hydrogen base on 6 of following second cycle line system,

Table 5 R ₁R ₂MP (℃) H SO ₂C ₄H ₉110-116

Table 6

N R ₁R ₂MP (℃) 1 H H 248-249

Claims

1. the chemical compound and the officinal salt thereof that have the following formula structure: Or Wherein Q is

Or Q contains 1,2 or 3 to be selected from the heteroatomic of N, O or S and to be unsubstituted or by R ⁸The 4-9 unit's monocycle or the bicyclo-ring system that replace; AB shares the adjacent carbon and the condensed ring system of nitrogen-atoms, wherein

B contains 1,2 or 3 heteroatomic 5,6 or 7 yuan of saturated or unsaturated ring that are selected from O, S or N; R ¹Be H, C _1-4Alkyl, N (R ⁸) ₂,-N (R ⁸) SO ₂R ⁷, NR ⁸CO ₂R ⁷, NR ⁸C (O) R ⁷, NR ⁸C (O) N (R ⁷) R ⁸, N (R ⁸) SO ₂N (R ⁷) R ⁸, N (R ⁸) SO ₂N (R ⁸) C (O) OR ⁷, C (O) N (R ⁷) ₂, or and R ⁶The cyclic group that forms, it is defined as follows; R ²Be H, C _1-4Alkyl, C _1-4Branched alkyl, C _1-4Alkylaryl, or aryl; R ⁵Be CH ,-CH (CH ₂) n, a key, probable back R ⁵Be contiguous N (R ⁴), R ⁶Be COOH, CH ₂OH, C (O) NR ⁷) ₂, CO ₂R ⁹, tetrazolium, the acyl group sulfonamide, or

, or and R ¹The cyclic group that forms, it is defined as follows; R wherein ¹With R ⁶The cyclic group that forms is

Wherein the alkyl of Qu Daiing is that hydroxyl replaces or C _1-4The alkyl that alkoxyl replaces, and the aryl that wherein replaces is replaced by one, two or three following groups: halogen, C _1-4Alkoxyl, hydroxyl or C _1-4Alkyl; R ⁸Be H, side chain or straight chain C _1-4Alkyl; R ⁹Be H, C _1-4Alkyl or aryl; N is 0-7; N ' is 0-3; With a be Or key.

2. the chemical compound and racemic modification, racemic mixture, enantiomer and the diastereomer that have the claim 1 of following formula structure: Or Wherein Q is

Or N=0-7; N ＇=0-3:R ⁴=H, C _1-4Alkyl, C _1-4Branched alkyl, ring-type C _1-4Alkyl or C _1-4Alkenyl; R ⁵=CH ,-CH (CH ₂) _n, or key; R ²Be H, C _1-4Alkyl, C _1-4Branched alkyl, C _1-4Alkylaryl, or aryl; R ¹=H, C _1-4Alkyl, N (R ⁸) ₂,-N (R ⁸) SO ₂R ⁷, NR ⁸CO ₂R ⁷, NR ⁸C (O) R ⁷, NR ⁸C (O) N (R ⁷) R ⁸, N (R ⁸) SO ₂N (R ⁷) R ⁸, N (R ⁸) SO ₂N (R ⁸) C (O) OR ⁷, C (O) N (R ⁷) ₂, or and R ⁶The cyclic group that forms; R ⁶=COOH, CH ₂OH, C (O) NR ⁷) ₂, CO ₂R ⁹, tetrazolium, the acyl group sulfonamide, or

Wherein Y=O or S:R ⁷=H, replacement or unsubstituted side chain or straight chain C _1-4Alkyl, side chain or straight chain low-grade alkenyl, C _1-4The aryl of alkylaryl, replacement or contain 1,2 or 3 N, S or the heteroatomic 5 or 6 yuan of heteroaryls of O,

Or key; A=contains 1,2 or 3 heteroatomic 5,6 or 7 yuan of saturated, the fractional saturations or the undersaturated ring that are selected from O, S or N; Contain 1,2 or 3 heteroatomic 5,6 or 7 yuan of saturated, fractional saturations or undersaturated ring that is selected from O, S or N with B=, wherein A and B form the condensed ring system of a shared adjacent carbons and nitrogen-atoms.

3. the chemical compound that has the claim 2 of following formula structure

Wherein AB is

Wherein V is N or CR ⁷And D is CH ₂, CH ₂-CH ₂, CH ₂C (R ⁷) ₂CH ₂, or

Wherein X=N or CR ³,

R wherein ³=CN, C (O) N (R ⁷) R ⁸, Or

With

4. the chemical compound of claim 3, wherein AB is

Wherein V is that N or CH and D are CH ₂-CH ₂Or CH ₂C (R ⁴) ₂CH ₂-

5. the chemical compound that has the claim 2 of following formula structure:

Wherein AB is Wherein V is N or CR ⁷With D be CH ₂, CH ₂-CH ₂, CH ₂C (R ⁷) ₂CH ₂, or

Wherein X=N or CR ³,

R wherein ³=CN, C (O) N (R ⁷) R ⁸,

Or

With

Y wherein ³Be O or H ₂

6. the chemical compound of claim 5, wherein a=

Be selected from AB

Wherein V is N or CR ⁷With D be CH ₂, CH ₂-CH ₂CH ₂C (R ⁷) ₂CH ₂Or

With Wherein X=N or CR ³, R wherein ³=CN, C (O) N (R ⁷) R ⁸,

Or

7. the chemical compound of claim 5, wherein key of a=; Be selected from AB

With

Y wherein ³Be O or H ₂

8. the chemical compound of claim 2, they are selected from

With

9. the chemical compound of claim 8, they are selected from:

With

10. be used for suppressing the mammal Fibrinogen and hematoblasticly combine, anticoagulant, treatment thrombosis or embolus forms or the chemical compound of the claim 2 that pre-preventing thrombosis or embolus form.

11. be used for suppressing the bonded compositions of mammal Fibrinogen and platelet, said composition comprises chemical compound and pharmaceutically suitable carrier of claim 2.

12. be used for suppressing the compositions of mammal platelet aggregation, said composition comprises chemical compound and pharmaceutically suitable carrier of claim 2.

13. be used for preventing the compositions of mammal thrombosis or embolus formation, said composition comprises chemical compound and pharmaceutically suitable carrier of claim 2.

14. be used for the treatment of the compositions that thrombosis in the mammal or embolus form, said composition comprises chemical compound and pharmaceutically suitable carrier of claim 2.

15. one kind is used for suppressing mammal Fibrinogen and the bonded method of platelet, this method comprises the compositions that goes up the claim 11 of effective dose to the administration pharmacology.

16. a method that suppresses platelet aggregation in the mammal, this method comprise the compositions that goes up the claim 12 of effective dose to the administration pharmacology.

17. a method that is used for preventing mammal thrombosis or embolus formation, this method comprises the compositions that goes up the claim 13 of effective dose to the administration pharmacology.

18. treat the method that thrombosis in the mammal or embolus form for one kind, this method comprises the compositions to administration claim 14.

19. a method that is used for suppressing the mammal platelet aggregation, this method comprise thrombolytic agent, claim 2 chemical compound and the anticoagulant of going up effective dose to the administration pharmacology.

20. the method for claim 19, wherein said thrombolytic agent are former activator of tissue plasminogen or streptokinase, and described anticoagulant is a heparin.

21. a method that is used for preventing or treating mammal thrombosis or embolus formation, this method comprises thrombolytic agent, claim 2 chemical compound of going up effective dose to the administration pharmacology, and anticoagulant.

22. the method for claim 21, wherein said thrombolytic agent are former activator of tissue plasminogen or streptokinase, and described anticoagulant is a heparin.