CN101010330A

CN101010330A - Partial and full agonists of A1 adenosine receptors

Info

Publication number: CN101010330A
Application number: CN 200580028768
Authority: CN
Inventors: 杰夫·扎布沃茨基; 埃尔法蒂赫·埃尔扎因; 迈克尔·G·奥根; 雅罗斯拉夫·比洛金; 斯坦尼斯拉斯·马耶尔; 安东尼·迪桑蒂; 斯科特·A·米勒; 彼得·A·克尔纳斯特
Original assignee: CV Therapeutics Inc
Current assignee: Gilead Palo Alto Inc
Priority date: 2004-08-30
Filing date: 2005-08-30
Publication date: 2007-08-01

Abstract

Disclosed are syntheses suitable for large scale manufacture of novel compounds that are partial and full A<SUB>1</SUB> adenosine receptor agonists, useful for treating various disease states, in particular tachycardia and atrial flutter, angina, and myocardial infarction.

Description

A 1The part of Adenosine Receptors and full agonist

The cross reference of related application

The U.S. Provisional Patent Application sequence number that the application requires to submit on August 30th, 2004 is 60/606,083 and be 60/622 in the U.S. Provisional Patent Application sequence number that on October 26th, 2004 submitted to, 076 right of priority is incorporated into this paper as a reference with the full content of its disclosure with way of reference.

Technical field

The present invention relates to partially or completely A of preparation conduct ₁The compound of adenosine receptor agonist.These compounds can be used for treating mammiferous diabetes, and obesity is improved the adipocyte function, CNS illness (obstacle), and improve the heart activity, especially treat irregular pulse.These compounds also have lipotropism and separate effect in mammalian body.

Background technology

Adenosine is a kind of naturally occurring nucleosides, its by be called A ₁, A _2A, A _2B, and A ₃The family of Adenosine Receptors interact and apply its biological effect, all these Adenosine Receptorss are all regulated the important physical process.For example, A _2AAdenosine Receptors is regulated coronary vasodilation, A _2BAcceptor then with adjusting, intestinal function and the neurocrine adjusting of mastocyte activation, asthma, vasodilation, cell growth (referring to Adenosine A _2BReceptors as Therapeutic Targets, Drug Dev Res 45:198; Feoktistov etal., Trends Pharmacol Sci 19:148-153), and A ₃It is relevant that Adenosine Receptors is regulated the cell proliferation process.

A ₁Two kinds of different physiological responses of Adenosine Receptors mediation.The inhibition of the cardiac stimulation effect of catecholamine is that the inhibition by means of adenylate cyclase mediates, and reducing heart rate (HR) and prolong the direct effect of propagating by the nerve impulse of AV knot and result from I to a great extent _KAdoActivation (B.Lerman and L.Belardinelli Circulation, Vol.83 (1991), P1499-1509 and J.C.Shryock and L.Belardinelli, The Am.J.Cardiology, Vol.79 (1997) P2-10).A ₁The stimulation of Adenosine Receptors can shorten the AV nodal cell action potential time length and reduce its scope, thereby prolong the refractory phase of AV nodal cell.Therefore, A ₁The stimulation of acceptor can provide treatment supraventricular tachycardic method, comprises stopping knot property reciprocal tachycardia, and controls Ventricular Rate in atrial fibrillation with during fluttering.

Therefore, A ₁Adenosine agonists can be used for treating the acute and chronic disease of the rhythm of the heart, and especially those are the disease of feature with quick heart rate, and wherein fast heart rate is to drive (activation) by unusual in antrum, atrium and the AV nodal tissue.Such illness includes but not limited to: atrial fibrillation, supraventricular tachycardia and auricular flutter.Be exposed to A ₁Agonist can cause that heart rate reduces and the adjustment of abnormal rhythm, thereby improves cardiovascular function.

Suppress the ability of catecholamine effect, A by it ₁Agonist can reduce the cAMP of cell, therefore has useful effect in the heart of depletion, and wherein the sympathetic tone of Zeng Jiaing can increase the cAMP level of cell.Research show, after illness follow the ventricular arrhythmia of increase and the possibility of sudden death.Referring to, for example, B.Lerman and L.Belardinelli Circulation, Vol.83 (1991), P1499-1509 and J.C.Shryock and L.Belardinelli, Am.J.Cardiology, Vol.79 (1997) P2-10.

Since the inhibiting result that it generates ring AMP, A ₁Agonist has lipotropism and separates effect in adipocyte, it causes reducing release (the E.A.van Schaick et al. of non-esterified fatty acid (NEFA), J.Pharmacokinetics and Biopharmaceutics, Vol.25 (1997) p 673-694 and P.Strong Clinical Science Vol.84 (1993) are p.663-669).Non-insulin-dependent diabetes mellitus (NIDDM) (NIDDM) is characterised in that insulin resistance, and it causes hyperglycemia.The factor of impelling viewed hyperglycemia is the activation that lacks normal glucose picked-up and skeletal muscle glycogen synthetase (GS).Studies show that the NEFA that improves the standard can suppress the glucose uptake and the glycogen synthetic (D.Thiebaud et al Metab.Clin.Exp.Vol.31 (1982) p1128-1136 and G.Boden et al J.Clin.Invest.Vol.93 (1994) p2438-2446) of insulin stimulating.As far back as P.J.Randle in 1963 glucose fat sour circulation hypothesis (P.J.Randle et al Lancet (1963) p.785-789) has just been proposed.The principle of this hypothesis is that restriction can promote carbohydrate utilization (P.Strong et al Clinical Science Vol.84 (1993) p.663-669) with the fatty acid supplying surrounding tissue.

Summarized A ₁(L.J.S.Knutsen and T.F.Murray are at Purinergic Approaches in ExperimentalTherapeutics for the benefit of nervus centralis illness (obstacle) for agonist, Eds.K.A.Jacobson and M.F.Jarvis (1997) Wiley-Liss, N.Y., P423-470).Briefly, the experimental model based on epilepsy studies show that, blended A _2A: A ₁Agonist (Y 341) is for by reverse benzodiazepine derivatives agonist 6,7-dimethoxy-4 '-ethyl-β-Ka Lin-3-carboxylate methyl ester institute inductive epileptic seizures is potent anticonvulsive drug (DMCM, H.Klitgaard Eur.J.Pharmacol. (1993) Vol.224p.221-228).Utilize CGS 21680 (a kind of A at other _2AAgonist) in the research of carrying out, draw following conclusion: anti-convulsant activity results from A ₁The activation of acceptor (G.Zhanget al.Eur.J.Pharmacol.Vol.255 (1994) p.239-243).In addition, studies show that A ₁The adenosine selective agonist has anti-convulsant activity (L.J.S.Knutsen In Adenosine and Adenne Nucleotides:From MolecularBiology to Integrative Physiology in the DMCM model; Eds.L.Belardinelli and A.Pelleg, Kluwer:Boston, 1995, pp479-487).A wherein ₁Second field that adenosine agonists has benefit is in the animal model of preceding cerebral ischemia, as by (J.Med.Chem.Vol.42 (1999) p.3463-3477) that the people proved such as Knutsen.Benefit in neuroprotective is considered to result partly to discharging the inhibition (ibid) of excitatory amino acid.

Verified, adenosine itself can effectively be treated and A ₁The illness that Adenosine Receptors is relevant for example stops paroxysmal supraventricular tachycardia.Yet, because the transformation period of adenosine is less than 10 seconds, so these effects are of short duration.In addition, because adenosine indistinguishably acts on A _2A, A _2B, and A ₃The Adenosine Receptors hypotype, so it also applies direct influence to sympathetic tone, coronary vasodilation, whole body vasodilation and mastocyte degranulation.

Reported that a class is as effective A ₁The compound of adenosine receptor agonist (fully and/or part) (be illustrated in the U.S. Patent application sequence submitted on July 17th, 2002 No. 10/194.335, its contents that all disclose are incorporated into this paper as a reference with way of reference).Studies show that, a kind of compound that in this patent application, discloses, its be confirmed as (4S, 5S, 2R, 3R)-5-[(2-fluorobenzene sulphur) methyl]-2-{6-[(2-hydroxyl-cyclopentyl) amino]-purine-9-yl Oxolan-3, the 4-glycol is the part A of high selectivity ₁Adenosine receptor agonist.

In view of showing great attention to this and similar compound, especially (4S, 5S, 2R, 3R)-and 5-[(2-fluorobenzene sulphur) methyl]-2-{6-[(2-hydroxyl-cyclopentyl) amino]-purine-9-yl } Oxolan-3, the diastereomer of 4-glycol, so wish to seek a kind of new synthetic method, the method that this synthetic method can be provided convenience prepares a large amount of good productive rate and highly purified above-claimed cpds and avoids using the separating step of chromatography and other effort.

Summary of the invention

Therefore, an object of the present invention is to provide a kind of synthetic method easily and come mass preparation (4S, 5S, 2R, 3R)-and 5-[(2-fluorobenzene sulphur) methyl]-2-{6-[(2-hydroxyl-cyclopentyl) amino]-purine-9-yl } Oxolan-3,4-two pure and mild related compounds, with and diastereomer, especially 2-{6-[((1S, 2S)-2-hydroxycyclopent base) amino] purine-9-yl } (4S, 5S, 3R)-and 5-[(2-fluorobenzene sulphur) methyl] Oxolan-3,4-glycol and 2-{6-[((1R, 2R)-2-hydroxycyclopent base) amino] purine-9-yl } (4S, 5S, 3R)-and 5-[(2-fluorobenzene sulphur) methyl] Oxolan-3, the 4-glycol.Therefore, aspect first, the present invention relates to prepare the compound of Formula I:

Wherein R is the phenyl of optional replacement:

Be included in that the compound with chemical formula RSH contacts with the compound of chemical formula (5) under the situation that alkali exists:

In a kind of preferred embodiment, R be 2-fluorophenyl and 6-substituting group be (1R, 2R)-2-(hydroxycyclopent base) amino.The example of the alkali that is fit to is sodium hydroxide, yellow soda ash, salt of wormwood, triethylamine, is preferably salt of wormwood, and reaction carries out in polar solvent usually, N for example, dinethylformamide or N,N-dimethylacetamide.

Aspect second, the present invention relates to prepare the compound of chemical formula (5), be included under the situation that alkali exists and contact with the compound of chemical formula (13) with 2-hydroxycyclopent base amine:

So that the compound of chemical formula (5) to be provided.

Reaction is carried out in protonic solvent usually, for example ethanol or Virahol, or methylene dichloride replacedly.The example of suitable alkali is sodium hydroxide, salt of wormwood and triethylamine, is preferably triethylamine.In a kind of preferred embodiment, 2-hydroxycyclopent base amine be as individual isomer be (1R, 2R)-2-hydroxycyclopent base amine exists.

Aspect the 3rd, the present invention relates to prepare the compound of chemical formula (13), comprise the compound that contacts chemical formula (12) with alkali;

Preferred alkali is ammoniacal liquor, and reacts and carry out in inert solvent usually, for example methyl alcohol or methylene dichloride.

Aspect the 4th, the present invention relates to prepare the compound of chemical formula (12), be included under the situation that alkali exists with thionyl chloride and 6-chloropurine nucleosides, promptly the compound of chemical formula (1) contacts:

In a kind of preferred embodiment, reaction is to carry out under the situation that inert solvent exists, for example acetonitrile or methylene dichloride.Alkali is pyridine normally.

Aspect the 5th, the present invention relates to prepare the compound of Formula I:

Wherein R is the phenyl of optional replacement:

May further comprise the steps:

(a) under the situation that alkali exists, contact with the compound of chemical formula (1) with thionyl chloride:

(b) use the compound of the product of alkali contact procedure (a) with formation chemical formula (13):

(c) under the situation that alkali exists, contact so that the compound of chemical formula (5) to be provided with the product of step (b) with 2-hydroxycyclopent base amine:

And

(d) compound with chemical formula RSH contacts with the product of step (c) under the situation that alkali exists, and wherein R as defined above.

Embodiment

Preferred embodiment

In step (a), reaction is carried out under the situation that inert solvent exists usually, for example acetonitrile or methylene dichloride.Alkali is pyridine normally.

In step (b), alkali is ammoniacal liquor normally.Reaction is preferably carried out under the situation that inert solvent exists.

In step (c), be reflected in the inert solvent and carry out, preferred alkali is triethylamine, and 2-hydroxycyclopent base amine preferably as individual isomer be (1R, 2R)-2-hydroxycyclopent base amine and existing.

In step (d), preferred solvent is N, dinethylformamide or N,N-dimethylacetamide, and N,N-dimethylacetamide more preferably, and preferred alkali is salt of wormwood.

Preferably, the product of the chemical formula in step (a) and the step (b), the compound of chemical formula (12) and chemical formula (13) is not separated, but is used as crude product in reactions steps thereafter.By this way, this method has been removed expensive and time-consuming purification step, and it is important when carrying out mass preparation and needs the problem of consideration.

Definition and general parameters

As employed in this manual, following word and phrase have implication as described below usually except that having in addition the explanation in the employed context therein.

Term " alkyl " is meant unit price side chain or unbranched saturated hydrocarbon chain, and it has 1 to 20 carbon atom.This term be wait by group such as methyl, ethyl, n-propyl, sec.-propyl, normal-butyl, isobutyl-, the tertiary butyl, n-hexyl, positive decyl, tetradecyl in addition illustrational.

Term " alkyl of replacement " is meant:

1) alkyl group as defined above; it has 1 to 5 substituting group; preferred 1 to 3 substituting group, these substituting groups are selected from by alkenyl; alkynyl; alkoxyl group; cycloalkyl; cycloalkenyl group; acyl group; amido; acyloxy; amino; aminocarboxyl; alkoxycarbonyl amino; azido-; cyano group; halogen; hydroxyl; ketone group; thiocarbonyl; carboxyl; carboxyalkyl; arylthio; heteroarylthio; the heterocycle sulfenyl; thiol; alkylthio; aryl; aryloxy; heteroaryl; amino-sulfonyl; amino carbonyl amino; heteroaryloxy; heterocyclic radical; heterocyclic oxy group; hydroxylamino; alkoxy amino; nitro;-SO-alkyl;-SO-aryl;-SO-heteroaryl;-SO ₂-alkyl ,-SO ₂-aryl and-SO ₂The group that-heteroaryl is formed.Unless by definition qualification is arranged in addition, all substituting groups can further be replaced by 1 to 3 substituting group alternatively, and these substituting groups are selected from alkyl, carboxyl, carboxyalkyl, aminocarboxyl, hydroxyl, alkoxyl group, halogen, CF ₃, amino, the amino that replaces, cyano group and-S (O) _nR, wherein R is alkyl, aryl or heteroaryl and n is 0,1 or 2; Or

2) alkyl group as defined above, its independently be selected from oxygen, sulphur and-NR _a-1 to 5 atom or group interrupt R wherein _aBe to be selected from hydrogen, alkyl, cycloalkyl, alkenyl, cycloalkenyl group, alkynyl, aryl, heteroaryl and heterocyclic radical.All substituting groups can be alternatively further by alkyl, alkoxyl group, halogen, CF ₃, amino, the amino that replaces, cyano group or-S (O) _nR replaces, and wherein R is alkyl, aryl or heteroaryl and n is 0,1 or 2; Or

3) alkyl group as defined above, it has as defined above 1 to 5 substituting group and by 1 to 5 atom or group interrupt as defined above.

Term " low alkyl group " is meant unit price side chain or unbranched saturated hydrocarbon chain, and it has 1 to 6 carbon atom.This term is to wait illustrational in addition by group such as methyl, ethyl, n-propyl, sec.-propyl, normal-butyl, isobutyl-, the tertiary butyl, n-hexyl.

Term " low alkyl group of replacement " is meant: low alkyl group as defined above, and it has as at defined 1 to 5 substituting group of alkyl that replaces, preferred 1 to 3 substituting group; Or low-grade alkyl group as defined above, its by as interrupted at defined 1 to 5 atom of alkyl that replaces; Or low-grade alkyl group as defined above, it has as defined above 1 to 5 substituting group and is interrupted by 1 to 5 atom as defined above.

Term " alkylidene group " is meant the double-basis of side chain or unbranched saturated hydrocarbon chain, and preferably it has 1 to 20 carbon atom, preferred 1 to 10 carbon atom, more preferably 1 to 6 carbon atom.This term is by group such as methylene radical (CH ₂-), ethylidene (CH ₂CH ₂-), the propylidene isomer (for example ,-CH ₂CH ₂CH ₂-and-CH (CH ₃) CH ₂-) wait illustrational in addition.

Term " low-grade alkylidene " is meant the double-basis of side chain or unbranched saturated hydrocarbon chain, and it has 1 to 6 carbon atom.

Term " alkylidene group of replacement " is meant:

(1) alkylidene group as defined above; it has 1 to 5 substituting group, and these substituting groups are selected from by alkyl; alkenyl; alkynyl; alkoxyl group; cycloalkyl; cycloalkenyl group; acyl group; amido; acyloxy; amino; aminocarboxyl; alkoxycarbonyl amino; azido-; cyano group; halogen; hydroxyl; ketone group; thiocarbonyl; carboxyl; carboxyalkyl; arylthio; heteroarylthio; the heterocycle sulfenyl; thiol; alkylthio; aryl; aryloxy; heteroaryl; amino-sulfonyl; amino carbonyl amino; heteroaryloxy; heterocyclic radical; heterocyclic oxy group; hydroxylamino; alkoxy amino; nitro;-SO-alkyl;-SO-aryl;-SO-heteroaryl;-SO ₂-alkyl ,-SO ₂-aryl and-SO ₂The group that-heteroaryl is formed.Unless by definition qualification is arranged in addition, all substituting groups can further be replaced by 1 to 3 substituting group alternatively, and these substituting groups are selected from alkyl, carboxyl, carboxyalkyl, aminocarboxyl, hydroxyl, alkoxyl group, halogen, CF ₃, amino, the amino that replaces, cyano group and-S (O) _nR, wherein R is alkyl, aryl or heteroaryl and n is 0,1 or 2; Or

(2) alkylidene group as defined above, it independently is selected from oxygen, sulphur and NR _a-1 to 5 atom or group interrupt R wherein _aBe alkyl, cycloalkyl, cycloalkenyl group, aryl, heteroaryl and the heterocyclic radical that is selected from hydrogen, optional replacement, or be selected from the group of carbonyl, carboxyl ester, carboxylic acid amides and alkylsulfonyl; Or

(3) alkylidene group as defined above, it has as defined above 1 to 5 substituting group and is interrupted by 1 to 20 atom as defined above.The example of the alkylidene group that replaces has chlorine methylene radical (CH (Cl)-), amino ethylidene (CH (NH ₂) CH ₂-), methylamino-ethylidene (CH (NHMe) CH ₂-), 2-carboxyl propylidene isomer (CH ₂CH (CO ₂H) CH ₂-), ethoxyethyl group (CH ₂CH ₂O-CH ₂CH ₂-), ethyl methylamino-ethyl (CH ₂CH ₂N (CH ₃) CH ₂CH ₂-), 1-oxyethyl group-2-(2-oxyethyl group-oxyethyl group) ethane (CH ₂CH ₂O-CH ₂CH ₂-OCH ₂CH ₂-OCH ₂CH ₂-) or the like.

Term " aralkyl " is meant the aromatic yl group that is covalently attached to alkylidene group, and wherein aryl and arylidene are defined in this article." aralkyl of optional replacement " is meant the aromatic yl group of the optional replacement of the alkylidene group that is covalently attached to optional replacement.Such aromatic alkyl group is illustrational in addition by benzyl, 3-(4-p-methoxy-phenyl) propyl group etc.

Term " alkoxyl group " is meant radicals R-O-, and wherein R is the alkyl of optional replacement or the cycloalkyl of optional replacement, or R is group-Y-Z, and wherein Y is the alkylidene group of optional replacement and Z is the alkenyl of optional replacement, the alkynyl of optional replacement; Or the cycloalkenyl group of optional replacement, wherein alkyl, alkenyl, alkynyl, cycloalkyl and cycloalkenyl group such as in this article definition.Preferred alkoxy base is alkyl-O-, and comprises, for example, and methoxyl group, oxyethyl group, positive propoxy, isopropoxy, n-butoxy, tert.-butoxy, sec-butoxy, n-pentyloxy, positive hexyloxy, 1,2-dimethyl butoxy or the like.

Term " alkylthio " is meant radicals R-S-, and wherein R is as defining at alkoxyl group.

Term " alkenyl " is meant the monoradical of side chain or unbranched unsaturated hydrocarbon group, preferably have 2 to 20 carbon atoms, more preferably 2 to 10 carbon atoms and even more preferably 2 to 6 carbon atoms, and have 1 to 6, preferred 1 two key (vinyl).Preferred kiki alkenyl group comprises vinyl (CH=CH ₂), 1-propylidene or allyl group (CH ₂CH=CH ₂), isopropylidene (C (CH ₃)=CH ₂), dicyclo [2.2.1] heptene or the like.Under alkenyl and situation that nitrogen links to each other, two keys can not be in the α position of nitrogen.

Term " low-grade alkenyl " is meant alkenyl as defined above, and it has 2 to 6 carbon atoms.

Term " alkenyl of replacement " is meant kiki alkenyl group as defined above; it has 1 to 5 substituting group; preferred 1 to 3 substituting group, these substituting groups are selected from by alkyl; alkenyl; alkynyl; alkoxyl group; cycloalkyl; cycloalkenyl group; acyl group; amido; acyloxy; amino; aminocarboxyl; alkoxycarbonyl amino; azido-; cyano group; halogen; hydroxyl; ketone group; thiocarbonyl; carboxyl; carboxyalkyl; arylthio; heteroarylthio; the heterocycle sulfenyl; thiol; alkylthio; aryl; aryloxy; heteroaryl; amino-sulfonyl; amino carbonyl amino; heteroaryloxy; heterocyclic radical; heterocyclic oxy group; hydroxylamino; alkoxy amino; nitro;-SO-alkyl;-SO-aryl;-SO-heteroaryl;-SO ₂-alkyl ,-SO ₂-aryl and-SO ₂The group that-heteroaryl is formed.Unless by definition qualification is arranged in addition, all substituting groups can further be replaced by 1 to 3 substituting group alternatively, and these substituting groups are selected from alkyl, carboxyl, carboxyalkyl, aminocarboxyl, hydroxyl, alkoxyl group, halogen, CF ₃, amino, the amino that replaces, cyano group and-S (O) _nR, wherein R is alkyl, aryl or heteroaryl and n is 0,1 or 2.

Term " alkynyl " is meant the monoradical of unsaturated hydrocarbons, preferably has 2 to 20 carbon atoms, more preferably 2 to 10 carbon atoms, even more preferably 2 to 6 carbon atoms, and have at least 1 and the unsaturated position of preferred 1 to 6 acetylene (triple bond).Preferred alkynyl group comprises ethynyl (C ≡ CH), propargyl (or proyl ,-C ≡ CCH ₃) or the like.Under alkynyl and situation that nitrogen links to each other, triple bond can not be in the α position of nitrogen.

Term " alkynyl of replacement " is meant alkynyl group as defined above; it has 1 to 5 substituting group; preferred 1 to 3 substituting group, these substituting groups are selected from by alkyl; alkenyl; alkynyl; alkoxyl group; cycloalkyl; cycloalkenyl group; acyl group; amido; acyloxy; amino; aminocarboxyl; alkoxycarbonyl amino; azido-; cyano group; halogen; hydroxyl; ketone group; thiocarbonyl; carboxyl; carboxyalkyl; arylthio; heteroarylthio; the heterocycle sulfenyl; thiol; alkylthio; aryl; aryloxy; heteroaryl; amino-sulfonyl; amino carbonyl amino; heteroaryloxy; heterocyclic radical; heterocyclic oxy group; hydroxylamino; alkoxy amino; nitro;-SO-alkyl;-SO-aryl;-SO-heteroaryl;-SO ₂-alkyl ,-SO ₂-aryl and-SO ₂The group that-heteroaryl is formed.Unless by definition qualification is arranged in addition, all substituting groups can further be replaced by 1 to 3 substituting group alternatively, and these substituting groups are selected from alkyl, carboxyl, carboxyalkyl, aminocarboxyl, hydroxyl, alkoxyl group, halogen, CF ₃, amino, the amino that replaces, cyano group and-S (O) _nR, wherein R is alkyl, aryl or heteroaryl and n is 0,1 or 2.

Term " aminocarboxyl " is meant group-C (O) NRR, and wherein each R is hydrogen, alkyl, aryl, heteroaryl, heterocyclic radical independently, or wherein two R groups link to each other to form heterocyclic group (for example, morpholino).All substituting groups can further be replaced by following radicals alternatively: alkyl, alkoxyl group, halogen, CF ₃, amino, the amino that replaces, cyano group or-S (O) _nR, wherein R is alkyl, aryl or heteroaryl and n is 0,1 or 2.

Term " amido " is meant group-NRC (O) R, and wherein each R is hydrogen, alkyl, aryl, heteroaryl or heterocyclic radical independently.All substituting groups can further be replaced by following radicals alternatively: alkyl, alkoxyl group, halogen, CF ₃, amino, the amino that replaces, cyano group or-S (O) _nR, wherein R is alkyl, aryl or heteroaryl and n is 0,1 or 2.

Term " acyloxy " be meant group-O (O) C-alkyl ,-O (O) C-cycloalkyl ,-O (O) C-aryl ,-O (O) C-heteroaryl and-O (O) C-heterocyclic radical.All substituting groups can further be replaced by following radicals alternatively: alkyl, alkoxyl group, halogen, CF ₃, amino, the amino that replaces, cyano group or-S (O) _nR, wherein R is alkyl, aryl or heteroaryl and n is 0,1 or 2.

Term " aryl " is meant the aromatic carbon ring group of 6 to 20 carbon atoms, and it has monocycle (for example, phenyl) or many rings (for example, xenyl), or a plurality of condensed ring (for example, naphthyl or anthryl).Preferred aryl groups comprises phenyl, naphthyl or the like.

Unless the definition at aryl substituent has qualification in addition; above-mentioned aromatic yl group can be alternatively by 1 to 5 substituting group; preferred 1 to 3 substituting group replaces, and these substituting groups are selected from by alkyl; alkenyl; alkynyl; alkoxyl group; cycloalkyl; cycloalkenyl group; acyl group; amido; acyloxy; amino; aminocarboxyl; alkoxycarbonyl amino; azido-; cyano group; halogen; hydroxyl; ketone group; thiocarbonyl; carboxyl; carboxyalkyl; arylthio; heteroarylthio; the heterocycle sulfenyl; thiol; alkylthio; aryl; aryloxy; heteroaryl; amino-sulfonyl; amino carbonyl amino; heteroaryloxy; heterocyclic radical; heterocyclic oxy group; hydroxylamino; alkoxy amino; nitro;-SO-alkyl;-SO-aryl;-SO-heteroaryl;-SO ₂-alkyl ,-SO ₂-aryl and-SO ₂The group that-heteroaryl is formed.Unless by definition qualification is arranged in addition, all substituting groups can further be replaced by 1 to 3 substituting group alternatively, and these substituting groups are selected from alkyl, carboxyl, carboxyalkyl, aminocarboxyl, hydroxyl, alkoxyl group, halogen, CF ₃, amino, the amino that replaces, cyano group and-S (O) _nR, wherein R is alkyl, aryl or heteroaryl and n is 0,1 or 2.

Term " aryloxy " is meant group aryl-O-, and wherein aromatic yl group and comprises the aromatic yl group of optional replacement equally as defined above as defined above.Term " arylthio " is meant radicals R-S-, and wherein R is as defining at aryl.

Term " amino " is meant group-NH ₂

Term " amino of replacement " is meant group-NRR, wherein each R by hydrogen, alkyl, cycloalkyl, carboxyalkyl (for example is independently selected from, benzyloxycarbonyl), the group of aryl, heteroaryl and heterocyclic radical composition, condition is that two R groups not all are hydrogen, or group-Y-Z, wherein Y is the alkylidene group of optional replacement and Z is alkenyl, cycloalkenyl group or alkynyl.Unless by definition qualification is arranged in addition, all substituting groups can further be replaced by 1 to 3 substituting group alternatively, and these substituting groups are selected from alkyl, carboxyl, carboxyalkyl, aminocarboxyl, hydroxyl, alkoxyl group, halogen, CF ₃, amino, the amino that replaces, cyano group and-S (O) _nR, wherein R is alkyl, aryl or heteroaryl and n is 0,1 or 2.

Term " carboxyalkyl " be meant group-C (O) O-alkyl ,-C (O) O-cycloalkyl, wherein alkyl and cycloalkyl are as defined in this article, and can further be replaced by following radicals alternatively: alkyl, alkenyl, alkynyl, alkoxyl group, halogen, CF ₃, amino, the amino that replaces, cyano group or-S (O) _nR, wherein R is alkyl, aryl or heteroaryl and n is 0,1 or 2.

Term " cycloalkyl " is meant the group of naphthene base of 3 to 20 carbon atoms, and it has monocycle or a plurality of condensed ring.Such group of naphthene base comprises, for example, and single ring architecture such as cyclopropyl, cyclobutyl, cyclopentyl, ring octyl group etc., or polynuclear plane such as adamantyl and dicyclo [2.2.1] heptane, or condense the group of naphthene base of aromatic yl group, indane for example, and class is with group.

Term " cycloalkyl of replacement " is meant group of naphthene base; it has 1 to 5 substituting group; preferred 1 to 3 substituting group, these substituting groups are selected from by alkyl; alkenyl; alkynyl; alkoxyl group; cycloalkyl; cycloalkenyl group; acyl group; amido; acyloxy; amino; aminocarboxyl; alkoxycarbonyl amino; azido-; cyano group; halogen; hydroxyl; ketone group; thiocarbonyl; carboxyl; carboxyalkyl; arylthio; heteroarylthio; the heterocycle sulfenyl; thiol; alkylthio; aryl; aryloxy; heteroaryl; amino-sulfonyl; amino carbonyl amino; heteroaryloxy; heterocyclic radical; heterocyclic oxy group; hydroxylamino; alkoxy amino; nitro;-SO-alkyl;-SO-aryl;-SO-heteroaryl;-SO ₂-alkyl ,-SO ₂-aryl and-SO ₂The group that-heteroaryl is formed.Unless by definition qualification is arranged in addition, all substituting groups can further be replaced by 1 to 3 substituting group alternatively, and these substituting groups are selected from alkyl, carboxyl, carboxyalkyl, aminocarboxyl, hydroxyl, alkoxyl group, halogen, CF ₃, amino, the amino that replaces, cyano group and-S (O) _nR, wherein R is alkyl, aryl or heteroaryl and n is 0,1 or 2.

Term " halogen " or " halo " are meant fluoro, bromo, chloro and iodo.

Term " acyl group " expression group-C (O) R, wherein R is the aryl of the heterocyclic radical of the cycloalkyl of the alkyl of hydrogen, optional replacement, optional replacement, optional replacement, optional replacement and the heteroaryl of optional replacement.

Term " heteroaryl " is meant aromatic group (promptly undersaturated), and it comprises 1 to 15 carbon atom and at 1 to 4 heteroatoms that is selected from oxygen, nitrogen and sulphur of at least one intra-annular.

Unless qualification is arranged in addition at the heteroaryl substituent definition; above-mentioned heteroaryl groups can be alternatively by 1 to 5 substituting group; preferred 1 to 3 substituting group replaces, and these substituting groups are selected from by alkyl; alkenyl; alkynyl; alkoxyl group; cycloalkyl; cycloalkenyl group; acyl group; amido; acyloxy; amino; aminocarboxyl; alkoxycarbonyl amino; azido-; cyano group; halogen; hydroxyl; ketone group; thiocarbonyl; carboxyl; carboxyalkyl; arylthio; heteroarylthio; the heterocycle sulfenyl; thiol; alkylthio; aryl; aryloxy; heteroaryl; amino-sulfonyl; amino carbonyl amino; heteroaryloxy; heterocyclic radical; heterocyclic oxy group; hydroxylamino; alkoxy amino; nitro;-SO-alkyl;-SO-aryl;-SO-heteroaryl;-SO ₂-alkyl ,-SO ₂-aryl and-SO ₂The group that-heteroaryl is formed.Unless by definition qualification is arranged in addition, all substituting groups can further be replaced by 1 to 3 substituting group alternatively, these substituting groups are selected from alkyl, carboxyl, carboxyalkyl, aminocarboxyl, hydroxyl, alkoxyl group, halogen, CF ₃, amino, the amino that replaces, cyano group and-S (O) _nR, wherein R is alkyl, aryl or heteroaryl and n is 0,1 or 2.Such heteroaryl groups can have monocycle (for example, pyridyl or furyl) or a plurality of condensed ring (for example, indolizine base, benzothiazolyl or benzothienyl).The example of nitrogen heterocyclic and heteroaryl includes but not limited to: pyrroles, imidazoles, pyrazoles, pyridine, pyrazine, pyrimidine, pyridazine, indolizine, isoindole, indoles, indazole, purine, quinolizine, isoquinoline 99.9, quinoline, 2 (phthalazines), naphthyl pyridine, quinoxaline, quinazoline, cinnolines (cinnoline), pteridine, carbazole, carboline, phenanthridines, acridine, phenanthroline, isothiazole, azophenlyene, isoxazole, phenoxazine, thiodiphenylamine, imidazolidine, tetrahydroglyoxaline, similar group and the heteroaryl compound that contains N-alkoxyl group-nitrogen.

Term " heteroaryloxy " is meant group heteroaryl-O-.

Term " heterocyclic radical " is meant the saturated or part unsaturated group of the unit price with monocycle or a plurality of condensed ring, and it has 1 to 40 carbon atom and at 1 to 10 heteroatoms that is selected from nitrogen, sulphur, phosphorus and/or oxygen of intra-annular, is preferably 1 to 4 heteroatoms.

The compound of Formula I comprises following definitions: " when considering with the nitrogen-atoms that links to each other with them, R and YR ¹The heterocyclic radical of representing optional replacement ".Such definition is included in the heterocycle that only contains nitrogen in the ring, for example, and pyrrolidines and piperidines, and be included in and have heteroatomic heterocycle more than, for example piperazines, morpholine class etc. in the ring.

Unless the definition at heterocyclic substituent has qualification in addition; above-mentioned heterocyclic group can be alternatively by 1 to 5 substituting group; preferred 1 to 3 substituting group replaces, and these substituting groups are selected from by alkyl; alkenyl; alkynyl; alkoxyl group; cycloalkyl; cycloalkenyl group; acyl group; amido; acyloxy; amino; aminocarboxyl; alkoxycarbonyl amino; azido-; cyano group; halogen; hydroxyl; ketone group; thiocarbonyl; carboxyl; carboxyalkyl; arylthio; heteroarylthio; the heterocycle sulfenyl; thiol; alkylthio; aryl; aryloxy; heteroaryl; amino-sulfonyl; amino carbonyl amino; heteroaryloxy; heterocyclic radical; heterocyclic oxy group; hydroxylamino; alkoxy amino; nitro;-SO-alkyl;-SO-aryl;-SO-heteroaryl;-SO ₂-alkyl ,-SO ₂-aryl and-SO ₂The group that-heteroaryl is formed.Unless by definition qualification is arranged in addition, all substituting groups can further be replaced by 1 to 3 substituting group alternatively, and these substituting groups are selected from alkyl, carboxyl, carboxyalkyl, aminocarboxyl, hydroxyl, alkoxyl group, halogen, CF ₃, amino, the amino that replaces, cyano group and-S (O) _nR, wherein R is alkyl, aryl or heteroaryl and n is 0,1 or 2.Heterocyclic group can have monocycle or a plurality of condensed ring.Preferred heterocycle comprises tetrahydrofuran base, morpholino, piperidyl etc.

Term " thiol " is meant group-SH.

Term " alkylthio of replacement " is meant the alkyl that group-S-replaces.

Term " assorted fragrant sulphur (hydroxyl) base " is meant group-S-heteroaryl, wherein heteroaryl groups as defined above, it comprises the heteroaryl groups of optional replacement equally as defined above.

Term " sulfoxide " is meant group-S (O) R, and wherein R is alkyl, aryl or heteroaryl." sulfoxide of replacement " is meant group-S (O) R, and wherein R is the alkyl as defined replacement in this article, the aryl of replacement or the heteroaryl that replaces.

Term " sulfone " is meant group-S (O) ₂R, wherein R is alkyl, aryl or heteroaryl." sulfone of replacement " is meant group-S (O) ₂R, wherein R is the alkyl as defined replacement in this article, the aryl of replacement or the heteroaryl that replaces.

Term " ketone group " be meant group-C (O)-.Term " thiocarbonyl " be meant group-C (S)-.Term " carboxyl " is meant group-C (O)-OH.

Term " protonic solvent " generally is meant the solvent that comprises oh group.The example of protonic solvent is methyl alcohol, ethanol, n-propyl alcohol, Virahol, butanols or the like.

" optionally " or " alternatively " is meant that thereafter incident or the situation described can take place or can not take place, and this description comprises the situation that situation that described incident or situation take place and described incident or situation do not take place.

Term " compound of Formula I " is the compound of the present invention that is used for comprising as disclosed, and the pharmaceutical salts of such compound, medicinal ester and prodrug.

Term " effectively therapeutic dose " is meant that the amount of the compound of Formula I is enough to be achieved as follows civilian defined effective treatment when to the Mammals administration of the such treatment of needs.Effectively therapeutic dose will change with following factor: the severity of the curee to be treated and the state of an illness, curee's body weight and age, the state of an illness, administering mode etc., these can be easy to determine by those of ordinary skills.

Term " treatment " or " processing " are meant any treatment to mammalian diseases, comprising:

(i) preventing disease promptly, does not develop the clinical symptom of disease;

(ii) suppress disease, that is, stop the development of clinical symptom; And/or

(iii) alleviate disease, that is, clinical symptom is disappeared.

In many cases, compound of the present invention is by the existence of amino and/or carboxylic group or group similar with it and can form acid salt and/or subsalt.Term " pharmaceutical salts " is meant such salt, and it keeps the biological effectiveness and the performance of the compound of Formula I, and on biology or in others is desirable.The medicinal basic additive salt can be by mineral alkali and organic bases preparation.Salt (only illustrating) derived from mineral alkali comprises sodium salt, sylvite, lithium salts, ammonium salt, calcium salt and magnesium salts.Salt derived from organic bases includes but not limited to: primary amine, the salt of secondary amine and tertiary amine, as alkylamine, dialkylamine, trialkylamine, the alkylamine that replaces, two (alkyl of replacement) amine, three (alkyl of replacement) amine, alkenyl amine, two alkenyl amines, three alkenyl amines, the alkenyl amine that replaces, two (alkenyl of replacement) amine, three (alkenyl of replacement) amine, Cycloalkyl amine, two (cycloalkyl) amine, three (cycloalkyl) amine, the Cycloalkyl amine that replaces, dibasic Cycloalkyl amine, trisubstituted Cycloalkyl amine, cycloalkenyl group amine, two (cycloalkenyl group) amine, three (cycloalkenyl group) amine, the cycloalkenyl group amine that replaces, dibasic cycloalkenyl group amine, trisubstituted cycloalkenyl group amine, arylamines, diarylamine, triarylamine, heteroaryl amine, two heteroaryl amine, three heteroaryl amine, heterocyclic radical amine, two heterocyclic radical amine, three heterocyclic radical amine, blended diamines and triamine, wherein at least two substituting groups on amine are different and are selected from by alkyl, the alkyl that replaces, alkenyl, the alkenyl that replaces, cycloalkyl, the cycloalkyl that replaces, cycloalkenyl group, the cycloalkenyl group that replaces, aryl, heteroaryl, heterocycle, and the group of similar group composition.Also comprise such amine, wherein two or three substituting groups and amino nitrogen form heterocyclic radical or heteroaryl groups together.

Only for illustrating, the particular instance of suitable amine comprises: Isopropylamine, Trimethylamine 99, diethylamine, tri-isopropyl amine, Tri-n-Propylamine, thanomin, 2-dimethylaminoethanol, Trometamol, Methionin, arginine, Histidine, caffeine, PROCAINE HCL, PHARMA GRADE, Hai Baming, choline, trimethyl-glycine, quadrol, glycosamine, N-alkylated glucamine, Theobromine, purine, piperazine, piperidines, morpholine, N-ethylpiperidine or the like.

Medicinal acid salt can be by mineral acid and organic acid preparation.Mineral acid that can salt derivative by it comprises hydrochloric acid, Hydrogen bromide, sulfuric acid, nitric acid, phosphoric acid etc.Organic acid that can salt derivative by it comprises acetate, propionic acid, oxyacetic acid, pyruvic acid, oxalic acid, oxysuccinic acid, propanedioic acid, Succinic Acid, toxilic acid, fumaric acid, tartrate, citric acid, phenylformic acid, styracin, amygdalic acid, methylsulfonic acid, ethyl sulfonic acid, tosic acid, Whitfield's ointment etc.

As used herein, " pharmaceutical carrier " comprises all solvents, dispersion medium, coating, antimicrobial drug and antifungal drug, isotonic agent and absorption delay agent or the like.Such medium and medicament are used for pharmaceutically active substance and are well known in the art.It is contemplated that and use it in the therapeutic composition, unless the medium of any routine or medicament and active ingredient are incompatible.Also auxiliary active ingredient can be added in the composition.

Compound as the agonist with highly inherent (intrinsic) effect can cause the maximum effect that biosystem can cause.These compounds are called " full agonist ".If it is higher to be incorporated into the efficient of effector process (effector process), they can bring out the effect of maximum possible and not occupy all acceptors.On the contrary, " partial agonist " can bring out reaction (replying), but can not bring out the maximum reaction (replying) that biosystem can cause.They can have rational avidity, but have lower specific efficacy.Part A ₁Adenosine agonists can have additional benefits for chronic treatment, because they lessly may induce A ₁The desensitization of acceptor (R.B.Clark, B.J.Knoll, R.Barber TiPS, Vol.20 (1999) are p.279-286), and lessly may cause side effect.

Name

The naming ﹠ numbering of compound of the present invention is that the typical compound with Formula I illustrates that wherein R is the 2-fluorophenyl:

It is named as:

(4S, 5S, 2R, 3R)-and 5-[(2-fluorophenyl sulfo-) methyl]-2-{6-[(2-hydroxycyclopent base) amino]-purine-9-yl } Oxolan-3, the 4-glycol, or:

2-{6-[((1RS, 2RS)-2-hydroxycyclopent base) amino] purine-9-yl } (4S, 5S, 3R)-and 5-[(2-fluorophenyl sulfo-) methyl] Oxolan-3, the 4-glycol.

Wherein the amino substituting group of 6-is derived from (1S, 2S)-related compound of 2-amino cyclopentyl-1-alcohol is named as 2-{6-[((1S, 2S)-2-hydroxycyclopent base) amino] purine-9-yl } (4S, 5S, 3R)-and 5-[(2-fluorophenyl sulfo-) methyl] Oxolan-3, the 4-glycol.Wherein the amino substituting group of 6-is derived from (1R, 2R)-related compound of 2-amino cyclopentyl-1-alcohol is named as 2-{6-[((1R, 2R)-2-hydroxycyclopent base) amino] purine-9-yl } (4S, 5S, 3R)-and 5-[(2-fluorophenyl sulfo-) methyl]-Oxolan-3, the 4-glycol.Wherein the amino substituting group of 6-is derived from (1R, 2S)-related compound of 2-amino cyclopentyl-1-alcohol is named as 2-{6-[((1R, 2S)-2-hydroxycyclopent base) amino] purine-9-yl } (4S, 5S, 3R)-and 5-[(2-fluorophenyl sulfo-) methyl]-Oxolan-3, the 4-glycol.Wherein the amino substituting group of 6-is derived from (1S, 2R)-related compound of 2-amino cyclopentyl-1-alcohol is named as 2-{6-[((1S, 2R)-2-hydroxycyclopent base) amino] purine-9-yl } (4S, 5S, 3R)-and 5-[(2-fluorophenyl sulfo-) methyl]-Oxolan-3, the 4-glycol.

The building-up reactions parameter

Term " solvent ", " inert organic solvents " or " inert solvent " are meant and are the inert solvent under the reaction conditionss of therewith describing, [comprise, for example, benzene, toluene, acetonitrile, tetrahydrofuran (THF) (" THF "), dimethyl formamide (" DMF "), chloroform, methylene dichloride, ether, methyl alcohol, pyridine etc.].Unless opposite regulation is arranged, employed solvent is an inert organic solvents in reaction of the present invention.

Term " capacity " is meant and adds the amount that is enough to reach described function, for example makes solution reach desirable volume (that is, 100%).

Synthesizing of the compound of Formula I

The process that is begun to prepare the compound of Formula I by the 6-chloropurine nucleosides has been shown in reaction scheme I.

Reaction scheme I

Wherein Ph is a phenyl.

The preparation of step 1-chemical formula (2)

The compound of chemical formula (2) is to react to make in about 24 hours in protonic solvent (preferred alcohol), at compound by chemical formula (1) and 2-(benzyloxy) cyclopentyl amine under the situation that alkali (preferred triethylamine) exists and under the temperature that refluxes approximately.When the completion of the reaction, come the product of separation chemistry formula (2) by ordinary method, for example, decompression removes down and desolvates, between ethyl acetate and water, distribute residue, from organic layer, remove and desolvate, and by for example coming the purifying residue by the crystallization or the precipitating action of ethyl acetate/hexane.

The preparation of step 2-chemical formula (3)

Then the compound of chemical formula (2) is changed into the compound of chemical formula (3).Under the situation that alkali (preferred pyridine) exists, thionyl chloride is joined in the suspension of compound in inert solvent (preferred acetonitrile) of chemical formula (2).Preferably in about 0 ℃ of down about 4 hours of reaction, make then to be warming up to room temperature and to spend the night.When the completion of the reaction, under reduced pressure the suspension that concentrates acquisition is to provide the compound of chemical formula (3), and it is used for next step and need not purifying.

The preparation of step 3-chemical formula (4)

The compound of chemical formula (4) is to make in the mixture of alkali (preferably ammonium hydroxide) and protonic solvent (particular methanol) by the compound dissolution with chemical formula (3).Reaction is about 30 minutes under about room temperature.When the completion of the reaction, come the product of separation chemistry formula (4) by ordinary method, for example, decompression removes down and desolvates, and distributes residue between ethyl acetate and water, and under reduced pressure removes ethyl acetate.Residue is used for next step and need not further purification.

The preparation of step 4-chemical formula (5)

Then under the situation that catalyzer (preferred palladium hydroxide) exists, by handling compound to go protection to chemical formula (4) with the unsaturated compound cycloalkyl of part (preferred tetrahydrobenzene).Replacedly, ammonium formiate can be used for replacing tetrahydrobenzene.In protonic solvent (preferred alcohol) and preferably, under about reflux temperature, reacted about 18 hours.When the completion of the reaction, come the product of separation chemistry formula (5), for example desolvate, then grind residue by under reduced pressure removing by ordinary method.

The preparation of step 5-Formula I

The compound of chemical formula (5) and the compound of chemical formula RSH (preferred 2-fluoro thiophenol) are reacted.Reaction is about 3 to 5 hours in protonic solvent (preferred N, dinethylformamide), under the situation that alkali (for example sodium hydroxide) exists and under about 100 ℃ temperature.Replacedly, reaction is about 1 to 5 day in protonic solvent (preferred N, dinethylformamide), under the situation that tertiary base (preferred triethylamine) exists and under about room temperature, preferred about 3 days.When the completion of the reaction, come the product of separation chemistry formula I, for example desolvate, then grind residue with ether by under reduced pressure removing by ordinary method.

The preparation of starting raw material

In step 1, with 1,2-(benzyloxy)-cyclopentyl amine is as starting raw material.This compound (as racemic mixture or as independent isomer) can commercially obtain maybe can be prepared by method well known to those skilled in the art.For example, and a kind of preparation (1R, 2R)-method of 2-(benzyloxy)-cyclopentyl amine is shown among the following reaction scheme II.

Reaction scheme II

In first step, by ordinary method, for example by in inert solvent and under the situation that the 4-Dimethylamino pyridine exists, reacting, with (BOC) ₂((1R, 2R)-2-amino cyclopentyl-1-alcohol) carries out the N protection to O (tert-Butyl dicarbonate) to the compound of chemical formula (a).Under the situation that alkali (preferred sodium hydride) exists, make the reaction of shielded cyclopentanol (b) derivative and bromotoluene then, in order to form (c), then the hydrochloric acid that for example is used in the dioxane with ordinary method goes protection to it.

From (1S, 2S)-the 2-amino cyclopentyl-beginning of 1-alcohol can provide the compound with stereochemistry characteristic opposite with chemical formula (d), and from (1RS, 2RS)-the pure beginning of 2-amino cyclopentyl-1-can provide the racemize analogue of the compound of chemical formula (d).

A kind of replaceable process that is used to prepare the compound of Formula I has been shown in reaction scheme III.

Reaction scheme III

Initial shielded cyclopentyl derivates can derived from (1R, 2R)-2-amino cyclopentyl-1-alcohol, (1S, 2S)-2-amino cyclopentyl-1-alcohol or (1RS, 2RS)-2-amino cyclopentyl-1-alcohol.By method well known in the art, the protected t-butyldimethylsilyl group that becomes of oh group.

Shown in reaction scheme IV, replacedly, compound that can conventional synthetic chemistry formula I and need not to use any blocking group.

Reaction scheme IV

A kind of method of compound of preferred preparation Formula I has been shown in reaction scheme V, and this method there is no need to utilize any blocking group or separation and/or purify intermediates.

Reaction scheme V

The preparation of step 1-chemical formula (12)

By the compound of chemical formula (1) being changed into the compound of chemical formula (12) with the reaction of thionyl chloride.Usually, under the situation that the alkali of about 2～2.5 molar equivalents (preferred pyridine) exists, the compound of chemical formula (1) is suspended in the inert solvent (preferred acetonitrile), and in about 1 hour, slowly adds the thionyl chloride of about 5～5.5 molar equivalents.Preferably in about 0 ℃ of down about 3 hours of reaction, make then to be warming up to room temperature and to spend the night.When the completion of the reaction, the suspension that under reduced pressure concentrates acquisition is to provide the compound of chemical formula (12), and it is preferred for next step and need not to purify.

The preparation of step 3-chemical formula (13)

Be dissolved in by crude product in the mixture of protonic solvent (preferred aqueous methanol) and alkali (preferably ammoniacal liquor), prepare the compound of chemical formula (13) by the compound of chemical formula (12) with step 1.Reacted about 1 hour down at about 0 ℃, then at room temperature reacted about 3 hours.When the completion of the reaction, by the product of ordinary method separation chemistry formula (13), and be used for next step and need not further purification.

The preparation of step 4-chemical formula (5)

In protonic solvent (preferred Virahol), under the situation that about 3 molar equivalent alkali (preferred triethylamine) exist, and under the temperature that refluxes approximately, reacted about 24 hours the compound of preparation chemical formula (5) by the crude product (compound of chemical formula (13)) of step 3 by 2-hydroxycyclopent base amine with about 1～1.1 molar equivalent.When the completion of the reaction, come the product of separation chemistry formula (5), for example remove and desolvate and blunge residue by reducing pressure down by ordinary method.

The preparation of step 5-Formula I

Make the product (compound of chemical formula (14)) of step 4 and the compound of the chemical formula RSH of about 3～5 molar equivalents then, for example 2-fluoro thiophenol reaction.In protonic solvent (being generally N, dinethylformamide), under the situation that the alkali of about 5～6 molar equivalents (for example sodium hydride, sodium hydroxide or triethylamine, preferred triethylamine) exists, reaction is about 1～5 day under about room temperature, preferably about 3 days.When the completion of the reaction, the product by ordinary method separation chemistry formula I.Can pass through by all kinds of SOLVENTS, for example the recrystallization of methyl alcohol, ethanol, Virahol or methyl alcohol and alcoholic acid mixture comes other purified product.Replacedly, can be by by the recrystallization of ethyl acetate or make slurry with ethyl acetate and come purified product.

Application, test and administration

The general application

The compound of Formula I can effectively be treated known to giving A ₁The respond illness of (replying) of the partially or completely agonist of Adenosine Receptors.Such illness includes but not limited to: the acute and chronic disease of the rhythm of the heart, and especially those are the disease of feature with quick heart rate, wherein (fast) heart rate is to drive (activation) by unusual in antrum, atrium and the AV nodal tissue.Such illness includes but not limited to: atrial fibrillation, supraventricular tachycardia and auricular flutter, congestive heart failure, non-insulin-dependent diabetes mellitus (NIDDM), hyperglycemia, epilepsy (anti-convulsant activity) and neuroprotective.A ₁Agonist also has lipotropism and separates effect in adipocyte, it causes reducing the release of non-esterified fatty acid.

Test

As described in, and carry out activity test by conspicuous method for a person skilled in the art at those patents cited above and document and following embodiment.

Following examples are to be used for illustrating preferred embodiment of the present invention.It should be understood by those skilled in the art that the technology representative that discloses in following examples is found by the inventor and in enforcement of the present invention the fine technology that works, therefore can be considered to constitute the optimal way of its enforcement.Yet the content that discloses according to the present invention it should be understood by those skilled in the art that under situation without departing from the spirit and scope of the present invention, can carry out multiple variation and still obtain identical or similar result disclosed specific embodiment.

Embodiment 1

The preparation of the compound of chemical formula (2)

To the 6-chloropurine nucleosides (10.0g, add in ethanol 35mmol) (350mL) solution triethylamine (10.0mL, 100mmol) and (1R, 2R)-2-(benzyloxy) cyclopentyl amine (5.2g, 52mmol).Backflow mixture 24 hours reacts from suspension during this period and becomes clear solution.Under reduced pressure remove ethanol, then at ethyl acetate and water (100mL: distribute residue 200mL).Separate organic layer, use ethyl acetate (2 * 75mL) washing water layers then.(sodium sulfate) dry organic layer that merges under reduced pressure removes then and desolvates.Residue is dissolved in the ethyl acetate (150mL), come precipitated product by adding hexane then, to obtain 2-(6-{[(1R as white solid, 2R)-and 2-(phenyl methoxyl group) cyclopentyl] amino } purine-9-yl) (4S, 3R, 5R)-and 5-(methylol) Oxolan-3,4-glycol (12.0g, 77%).

¹H?NMR(CD ₃OD)δ1.62-2.16(m，6H)，3.26-3.29(m，1H，NHC H)，3.68-3.85(m，2H，CH ₂-5’)，4.03-4.10(m，1H，CH-4’)，4.12-4.16(m，1H，CHOBn)，4.16-4.19(m，1H，3’CH)，4.71(s，2H，OCH ₂Ph)，4.83-4.92(m，1H，2’CH)，5.98(d，J＝6Hz，1H，H-1’)，7.23-7.35(m，5H，PhH)，8.15(S，1H，C-2H)。

B. The preparation of the compound of chemical formula (2)

Similarly, according to the step of above 1A, but with other isomer of 2-(benzyloxy) cyclopentyl amine replace (1R 2R)-2-(benzyloxy) cyclopentyl amine, makes following compound:

2-(6-{[(1S, 2S)-2-(phenyl methoxyl group) cyclopentyl] amino } purine-9-yl) (4S, 3R, 5R)-and 5-(methylol) Oxolan-3, the 4-glycol;

2-(6-{[(1R, 2S)-2-(phenyl methoxyl group) cyclopentyl] amino } purine-9-yl) (4S, 3R, 5R)-and 5-(methylol) Oxolan-3, the 4-glycol;

2-(6-{[(1S, 2R)-2-(phenyl methoxyl group) cyclopentyl] amino } purine-9-yl) (4S, 3R, 5R)-and 5-(methylol) Oxolan-3, the 4-glycol; And

2-(6-{[(1RS, 2RS)-2-(phenyl methoxyl group) cyclopentyl] amino } purine-9-yl) (4S, 3R, 5R)-and 5-(methylol) Oxolan-3, the 4-glycol.

Embodiment 2

The preparation of the compound of chemical formula (3)

Under 0 ℃, to the 2-(6-{[(1R in acetonitrile (15mL), 2R)-and 2-(phenyl methoxyl group) cyclopentyl] amino } purine-9-yl) (4S, 3R, 5R)-5-(methylol) Oxolan-3, the 4-glycol (2.0g, 4.5mmol) and pyridine (0.728mL, 9mmol) thionyl chloride in the suspension through stirring (1.7mL, 22.5mmol).After stirring 4 hours under 0 ℃, make reaction be warming up to room temperature, stir then and spend the night.Under reduced pressure remove and desolvate from the suspension that produces, with provide 4-(6-{[(1R, 2R)-2-(phenyl methoxyl group) cyclopentyl] amino purine-9-yl) (6S, 3aR, 6aR)-6-(chloromethyl)-4H, 6H, 3 aH, 6aH-Oxolan also [3,4-d] 1,3,2-two oxa-thiacyclopentane (dioxathiolan)-2-ketone, it is used for next step and need not further purification.

B. The preparation of the compound of chemical formula (3)

Similarly, step according to above 2A, but with 2-(6-{[2-(phenyl methoxyl group) cyclopentyl]-amino purine-9-yl) (4S, 3R, 5R)-5-(methylol) Oxolan-3, other isomer of 4-glycol replaces 2-(6-{[(1R, 2R)-2-(phenyl methoxyl group) cyclopentyl]-amino } purine-9-yl) (4S, 3R, 5R)-5-(methylol) Oxolan-3, the 4-glycol makes following compound:

4-(6-{[(1S, 2S)-2-(phenyl methoxyl group) cyclopentyl] amino } purine-9-yl) (6S, 3aR, 6aR)-and 6-(chloromethyl)-4H, 6H, 3aH, 6aH-Oxolan be [3,4-d] 1,3 also, 2-two oxa-s thiacyclopentane-2-ketone;

4-(6-{[(1R, 2S)-2-(phenyl methoxyl group) cyclopentyl] amino } purine-9-yl) (6S, 3aR, 6aR)-and 6-(chloromethyl)-4H, 6H, 3aH, 6aH-Oxolan be [3,4-d] 1,3 also, 2-two oxa-s thiacyclopentane-2-ketone;

4-(6-{[(1S, 2R)-2-(phenyl methoxyl group) cyclopentyl] amino } purine-9-yl) (6S, 3aR, 6aR)-and 6-(chloromethyl)-4H, 6H, 3aH, 6aH-Oxolan be [3,4-d] 1,3 also, 2-two oxa-s thiacyclopentane-2-ketone; And

4-(6-{[(1RS, 2RS)-2-(phenyl methoxyl group) cyclopentyl] amino } purine-9-yl) (6S, 3aR, 6aR)-and 6-(chloromethyl)-4H, 6H, 3aH, 6aH-Oxolan be [3,4-d] 1,3 also, 2-two oxa-s thiacyclopentane-2-ketone.

Embodiment 3

The preparation of the compound of chemical formula (4)

Will be from the 4-(6-{[(1R of embodiment 2,2R)-and 2-(phenyl methoxyl group) cyclopentyl] amino } purine-9-yl) (6S, 3aR, 6aR)-6-(chloromethyl)-4H, 6H, 3aH, 6aH-Oxolan be [3,4-d] 1 also, 3,2-two oxa-s thiacyclopentane-2-ketone is dissolved in the mixture of first alcohol and water (40mL/2mL), drips dense ammonium hydroxide (2.2mL, 28%) then in this solution.Stirring is after 30 minutes down at 23 ℃, and under reduced pressure except that desolvating, water (15mL) dilutes residue then.(3 * 75mL) extraction aqueous mixtures are used MgSO with ethyl acetate ₄Drying is under reduced pressure removed then and is desolvated, with provide 2-(6-{[(1R, 2R)-2-(phenyl methoxyl group) cyclopentyl] amino purine-9-yl) (4S, 5S, 3R)-5-(chloromethyl) Oxolan-3, the 4-glycol, it is used for next step and need not further purification.

B. The preparation of the compound of chemical formula (4)

Similarly, according to the step of above 3A, but with 4-(6-{[2-(phenyl methoxyl group) cyclopentyl] amino } purine-9-yl) (6S, 3aR, 6aR)-and 6-(chloromethyl)-4H, 6H, 3aH, 6aH-Oxolan also [3,4-d] 1,3, other isomer of 2-two oxa-s thiacyclopentane-2-ketone replaces 4-(6-{[(1R, 2R)-and 2-(phenyl methoxyl group) cyclopentyl] amino } purine-9-yl) (6S, 3aR, 6aR)-6-(chloromethyl)-4H, 6H, 3aH, the 6aH-Oxolan is [3,4-d] 1,3 also, 2-two oxa-s thiacyclopentane-2-ketone makes following compound:

2-(6-{[(1S, 2S)-2-(phenyl methoxyl group) cyclopentyl] amino } purine-9-yl) (4S, 5S, 3R)-and 5-(chloromethyl) Oxolan-3, the 4-glycol;

2-(6-{[(1R, 2S)-2-(phenyl methoxyl group) cyclopentyl] amino } purine-9-yl) (4S, 5S, 3R)-and 5-(chloromethyl) Oxolan-3, the 4-glycol;

2-(6-{[(1S, 2R)-2-(phenyl methoxyl group) cyclopentyl] amino } purine-9-yl) (4S, 5S, 3R)-and 5-(chloromethyl) Oxolan-3, the 4-glycol; And

2-(6-{[(1RS, 2RS)-2-(phenyl methoxyl group) cyclopentyl] amino } purine-9-yl) (4S, 5S, 3R)-and 5-(chloromethyl) Oxolan-3, the 4-glycol.

Example 4

The preparation of the compound of chemical formula (5)

The 2-that will in embodiment 3, obtain (6-{[(1R, 2R)-2-(phenyl methoxyl group) cyclopentyl] amino } purine-9-yl) (4S, 5S, 3R)-and 5-(chloromethyl) Oxolan-3,4-glycol (22g) is dissolved in ethanol (450mL) and the hexanaphthene (200mL).In this solution, add palladium hydroxide (20 moles of % add 1 gram at first, add 1 gram after 6 hours, and add 1 gram after 14 hours), and made reaction mixture refluxed 18 hours.Reaction mixture is still being filtered by diatomite (or Yin Shi salt) in the heat, under reduced pressure from filtrate, removing then and desolvate.With ethanol (20mL) grinding product, filter, use washing with alcohol then, so that the 2-{6-[((1R as white powder to be provided, 2R)-and 2-hydroxycyclopent base) amino] purine-9-yl } (4S, 5S, 3R)-and 5-(chloromethyl) Oxolan-3,4-glycol (7.3 gram).

In addition, by callable palladium hydroxide is suspended in the methyl alcohol (200mL), and came salvage material in 1 hour at 90 ℃ of following heated mixt.By the diatomite filtration hot mixt, further wash diatomite with hot methanol then.Under reduced pressure concentrated filtrate uses ethanol (20mL) to grind residue then, so that the 2-{6-[((1R of other 8.6 grams to be provided, 2R)-and 2-hydroxycyclopent base) amino] purine-9-yl } (4S, 5S, 3R)-and 5-(chloromethyl) Oxolan-3, the 4-glycol.

¹H?NMR(DMSO-d6)δ1.64-2.18(m，6H)，3.26-3.29(m，1H，NHC H)，3.83-3.97(m，2H，CH ₂Cl?5’)，4.03-4.09(m，1H，CH-4’)，4.12-4.17(m，1H，CHOH)，4.16-4.19(m，1H，3’CH)，4.84-4.92(m，1H，2’CH)，5.96(d，J＝6Hz，1H，H-1’)，7.23-7.35(m，5H，PhH)，8.15(S，1H，C-2H)，8.39(s，1H，C-8H)。

B. The preparation of the compound of chemical formula (5)

Similarly, step according to above 4A, but with 2-(6-{[2-(phenyl methoxyl group) cyclopentyl] amino } purine-9-yl) (4S, 5S, 3R)-5-(chloromethyl) Oxolan-3, other isomer of 4-glycol replaces 2-(6-{[(1R, 2R)-and 2-(phenyl methoxyl group) cyclopentyl] amino } purine-9-yl) (4S, 5S, 3R)-5-(chloromethyl) Oxolan-3, the 4-glycol makes following compound:

Embodiment 5

The preparation of the compound of Formula I, wherein R is the 2-fluorophenyl

To 2-fluoro thiophenol (38mL, be added in N in 2N sodium hydroxide (100mL) solution 406mmol), 2-{6-[((1R in the dinethylformamide (120mL), 2R)-and 2-hydroxycyclopent base) amino] purine-9-yl } (4S, 5S, 3R)-5-(chloromethyl) Oxolan-3, the 4-glycol (15.0g, 40.6mmol).Mixture is warming up to 100 ℃ continues 4 hours, follow the tracks of reaction process by TLC.Under reduced pressure remove N, dinethylformamide, then water (200mL) dilutes remaining mixture, and with the acetate neutralization, (MgSO is used in 3 * 125mL) extractions then with ethyl acetate ₄The dry organic layer that merges.Under reduced pressure, grind residue and filtration, so that the 2-{6-[((1Rs of 16 grams as white powder to be provided with ether except that after desolvating, 2R)-and 2-hydroxycyclopent base) amino] purine-9-yl } (4S, 5S, 3R)-5-[(2-fluorophenyl sulfo-) methyl] Oxolan-3,4-glycol (productive rate is 85%).

¹H?NMR(DMSO-d6)δ1.66-2.27(m，6H)，3.42-3.59(m，1H，NHC H)，4.05-4.14(m，2H)，4.03-4.09(m，1H，CH-4’)，4.14-4.19(m，1H)，4.16-4.19(m，1H，3’CH)，4.84-4.92(m，1H，2’CH)，5.97(d，J＝6Hz，1H，H-1’)，7.05-7.55(m，4H，PhH)，8.10(S，1H，C-2H)，8.15(s，1H，C-8H)。

B. The preparation of the compound of Formula I, wherein R is the 2-fluorophenyl

Similarly, step according to above 5A, but with 2-{6-[(2-hydroxycyclopent base) amino] purine-9-yl } (4S, 5S, 3R)-5-(chloromethyl) Oxolan-3, other isomer of 4-glycol replaces 2-{6-[((1R, 2R)-and 2-hydroxycyclopent base) amino] purine-9-yl } (4S, 5S, 3R)-5-(chloromethyl) Oxolan-3, the 4-glycol makes following compound:

2-{6-[((1S, 2S)-2-hydroxycyclopent base) amino] purine-9-yl } (4S, 5S, 3R)-and 5-[(2-fluorophenyl sulfo-) methyl] Oxolan-3, the 4-glycol;

2-{6-[((1R, 2S)-2-hydroxycyclopent base) amino] purine-9-yl } (4S, 5S, 3R)-and 5-[(2-fluorophenyl sulfo-) methyl] Oxolan-3, the 4-glycol;

2-{6-[((1S, 2R)-2-hydroxycyclopent base) amino] purine-9-yl } (4S, 5S, 3R)-and 5-[(2-fluorophenyl sulfo-) methyl] Oxolan-3, the 4-glycol; And

C. The preparation of the compound of Formula I wherein changes R

Similarly,, but replace the 2-fluoro thiophenol, make other compound of Formula I with other thiophenol of chemical formula RSH according to the step of above 5A.

Embodiment 6

The preparation of the compound of chemical formula (12)

Preparation 1

In 55 minutes time, (50.0g is 174.4mmol) with through distillatory pyridine (30ml, thionyl chloride (SOCl in cold (0 ℃, ice bath) suspension 370mmol) to the 6-chloropurine nucleosides in exsiccant acetonitrile (600ml) ₂, 66.0ml, 907mmol).0 ℃ of following stirred reaction mixture 3 hours, at room temperature stirred then 18 hours.At the following concentrated yellow solutions of decompression and 40 ℃, drying 6 hours under high vacuum then.With residue (6S, 4R, 3aR, 6aR)-6-(chloromethyl)-4-(6-chloropurine-9-yl)-4H, 6H, 3aH, 6aH-Oxolan be [3,4-d] 1,3 also, 2-two oxa-s thiacyclopentane-2-ketone (12) is used for subsequent reaction and need not further purification.

2. The interchangeable preparation of the compound of chemical formula (12)

In 30～60 minutes time, to the 6-chloropurine nucleosides (1Kg) in exsiccant methylene dichloride (15 liters) and in the mixture of distillatory pyridine (850ml) thionyl chloride (SOCl ₂, 530ml), keep temperature to be lower than 30 ℃ simultaneously.30 ℃ of following stirred reaction mixtures 4 hours, be cooled to 20 ℃ then.Add dehydrated alcohol (1700ml), maintain the temperature at 20 ℃, and stirred the mixture 15 minutes.Slowly add entry (3.5 liters) then, and stirred the mixture 30 minutes, thereafter the separate content thing.Be separated, use saturated sodium bicarbonate (4 liters) washing organic layer then.Two be separated after, with saturated sodium-chloride (2.6 liters) washing organic layer, separate, under reduced pressure remove then and desolvate until reaching about 4 liters volume, thereby (6S, the 4R of dissolved state are provided, 3aR, 6aR)-6-(chloromethyl)-4-(6-chloropurine-9-yl)-4H, 6H, 3aH, the 6aH-Oxolan is [3,4-d] 1,3 also, the solution of 2-two oxa-s thiacyclopentane-2-ketone (12), it is used for subsequent reaction and need not further purification.

Embodiment 7

The preparation of the compound of chemical formula (13)

Will be in methyl alcohol (1000ml) and distilled water (50ml) available from the compound dissolution of the chemical formula (12) of embodiment 6 (preparation 1).Solution is cooled to 0 ℃, in 25 minutes, drip then strong aqua (28%, 56ml).0 ℃ of following continuously stirring 1 hour, at room temperature stirred then 3 hours.The strong aqua (28%) that adds other 10ml during this period (is followed the tracks of reaction process, CH by TLC ₂Cl ₂/ MeOH is 9: 1).Concentrated reaction mixture and at room temperature use 5% aqueous solution (1000ml) the hydrolysis residue of citric acid under reduced pressure then.With ethyl acetate (1 * 900ml, 1 * 400ml, 1 * 200ml, 2 * 100ml) aqueous layer extracted, the organic layer of using saturated sodium bicarbonate (450ml) washing to merge then.With ethyl acetate (3 * 50ml) extraction sodium bicarbonate water layers.With the organic layer that salt solution (400ml) washing merges, use ethyl acetate (3 * 50ml) extraction sodium-chlor water layers then.Organic layer with dried over sodium sulfate merges filters, then under reduced pressure concentrated filtrate with the compound that provides 41.8g chemical formula (13) (4S, 5S, 2R, 3R)-5-(chloromethyl)-2-(6-chloropurine-9-yl) Oxolan-3, the 4-glycol.Further purify.

Preparation 2

Replacedly, (6S, 4R, 3aR to the dissolved state that in embodiment 6 preparations 2, obtains, 6aR)-6-(chloromethyl)-4-(6-chloropurine-9-yl)-4H, 6H, 3aH, 6aH-Oxolan also [3,4-d] 1,3, add ammonium hydroxide (500ml) in the solution of 2-two oxa-s thiacyclopentane-2-ketone (12), under 25 ℃, stirred the mixture 12 hours then.Leach solid, use methylene dichloride (500ml) washing then.Merging filtrate and washes are reduced to about 6 liters with volume then under vacuum.Further purify.

Embodiment 8

The preparation of the compound of chemical formula (5)

Preparation 1

To (the R in degassing Virahol (100ml), R)-amino amylalcohol hydrochloride (or hydrochloride) (34.2g of 2-, 249mmol) (use the hydrolith drying, 95ml, 69g with the distillation triethylamine, drip (the 4S in the 400ml Virahol in suspension 226mmol), 5S, 2R, 3R)-5-(chloromethyl)-2-(6-chloropurine-9-yl) Oxolan-3,4-glycol (36.3g, 118.7mmol) solution.At room temperature stirred reaction mixture is 30 minutes, and backflow (about 80 ℃ of oil bath temperature) is 20 hours then.After reaction mixture is cooled to room temperature, under reduced pressure remove and desolvate, in residue, add 1000ml water then.At room temperature stirred suspension is 3.5 hours, leaches solid matter subsequently, washes that (1 * 60ml and 1 * 90ml) uses P then under vacuum with water ₂O ₅Dry 3 days to provide 68.0g (81%) 2-{6-[((1R as light brown powder, 2R)-2-hydroxycyclopent base) amino] purine-9-yl } (4S, 5S, 3R)-and 5-(chloromethyl) Oxolan-3, the 4-glycol.

Preparation 2

Replacedly, the solution that will obtain in embodiment 7, preparation 2 is cooled to 20～25 ℃, adds triethylamine (1000ml) then, then add (R, R)-the amino amylalcohol (530g) of 2-.Backflow mixture 8 hours under atmospheric pressure removes then and desolvates until reaching about 4 liters volume.Mixture is cooled to 55～60 ℃, adds entry (15 liters), mixture is cooled to 20～25 ℃ then.Stirred the mixture about 1 hour, and filtered then, with dehydrated alcohol (1.25 liters) washing solid, drying under reduced pressure solid subsequently, therebetween not allowable temperature above 60 ℃.

B. similarly, step according to above 8A (preparation 1 or prepare 2), but with (S, S)-and the amino amylalcohol hydrochloride of 2-(or hydrochloride) replacement (R, R)-the amino amylalcohol hydrochloride (or hydrochloride) of 2-, then make 2-{6-[((1S, 2S)-and 2-hydroxycyclopent base) amino] purine-9-yl } (4S, 5S, 3R)-5-(chloromethyl) Oxolan-3, the 4-glycol.

C. similarly, step according to above 8A (preparation 1 or prepare 2), but with (1R, 2S)-and the amino amylalcohol hydrochloride of 2-(or hydrochloride) replacement (R, R)-the amino amylalcohol hydrochloride (or hydrochloride) of 2-, then make 2-{6-[((1R, 2S)-and 2-hydroxycyclopent base) amino] purine-9-yl } (4S, 5S, 3R)-5-(chloromethyl) Oxolan-3, the 4-glycol.

D. similarly, step according to above 8A (preparation 1 or prepare 2), but with (1S, 2R)-and the amino amylalcohol hydrochloride replacement of 2-(R, R)-the amino amylalcohol hydrochloride of 2-, then make 2-{6-[((1S, 2R)-and 2-hydroxycyclopent base) amino] purine-9-yl } (4S, 5S, 3R)-5-(chloromethyl) Oxolan-3, the 4-glycol.

Embodiment 9

The preparation of the compound of Formula I, wherein R is the 2-fluorophenyl

Preparation 1

To the degassing anhydrous N, the 2-{6-[((1R in the dinethylformamide (1.8 liters), 2R)-2-hydroxycyclopent base) amino]-purine-9-yl (4S, 5S, 3R)-5-(chloromethyl) Oxolan-3,4-glycol (166.5g, 0.457mol) and by hydrolith distillatory triethylamine (352ml, 256g, 2.53mol, 4 equivalents) solution in add 2-fluoro thiophenol (190ml, 228g, 1.78mol, 4 equivalents), the part of every 2-3 hour adding 38.5ml.At room temperature stirred the mixture 4 days, continuously nitrogen is blasted simultaneously in the solution (by ¹H NMR monitors reaction).After reaction is finished, reaction mixture is injected in 7 liters the ethyl acetate, wash with 3 premium on currency then.(2 * 500ml) aqueous layer extracted, and the organic layer of water (3 * 2 liters) washing merging are reduced to about 1.8 liters volume then, thereby the suspension of white solid are provided with ethyl acetate.At room temperature stirred suspension is 9 hours, leach white depositions then, with ether (3 * 200ml) washings, then under high vacuum dry 24 hours, to provide the 2-{6-[((1R of 131g (productive rate is 63%) as white powder, 2R)-2-hydroxycyclopent base) amino] purine-9-yl } (4S, 5S, 3R)-and 5-[(2-fluorophenyl sulfo-) methyl]-Oxolan-3,4-glycol (purity is 98.9%).

By in 1 liter of ether (50: 1), stirring to spend the night product is further purified, to provide the pure 2-{6-[((1R of 127g, 2R)-2-hydroxycyclopent base) amino] purine-9-yl } (4S, 5S, 3R)-and 5-[(2-fluorophenyl sulfo-) methyl]-Oxolan-3, the 4-glycol.

Preparation 2

The product (1Kg) of embodiment 8, preparation 2 is dissolved in the N,N-dimethylacetamide (2.7 liters), adds salt of wormwood (560g) then.In keeping below 25 ℃ mixture, add 2-fluoro thiophenol (380g), then about 6 hours of 60～65 ℃ of following heated mixt.Mixture is cooled to 25～30 ℃ then, and adds ethyl acetate (10 liters), then add the solution of the water (4.9 liters) of sodium-chlor (260g), and stirred the mixture 15 minutes.Be separated into two-layer after, wash organic phase once more with water (4.9 liters) solution of sodium-chlor (260g), stirred the mixture then 15 minutes.After separating, under atmospheric pressure organic layer is concentrated into about 5 liters volume, add methyl alcohol (10 liters) then.Under atmospheric pressure once more mixture is concentrated into about 2.8 liters volume, then the solution that obtains is cooled to about 35～40 ℃.Add methylene dichloride (5 liters) then, and with mixture be maintained at about 35～40 ℃ 1 hour, then be cooled between 0～5 ℃ 30 minutes.Leach solid product, wash with methylene dichloride (2.8 liters), drying under reduced pressure is to constant weight then, allowable temperature is not higher than 50 ℃ therebetween, so that 2-{6-[((1R to be provided, 2R)-2-hydroxycyclopent base) amino] purine-9-yl } (4S, 5S, 3R)-and 5-[(2-fluorophenyl sulfo-) methyl]-Oxolan-3, the 4-glycol.

By following step product is further purified: under the temperature between 60～70 ℃, product (2-{6-[((1R with 1Kg, 2R)-and 2-hydroxycyclopent base) amino] purine-9-yl } (4S, 5S, 3R)-5-[(2-fluorophenyl sulfo-) methyl]-Oxolan-3, the 4-glycol) be dissolved in the methyl alcohol (20 liters), be incubated 1 hour, be cooled to 45～50 ℃, pass through 1 micron strainer filtering solution then, and keep solution temperature to be higher than 40 ℃.To about 7 liters, and keep solution temperature to be higher than 40 ℃ solution concentration, then the solution that obtains is remained on 50～55 ℃ 1 hour.In 2 hours, solution is cooled to-5 ℃ then, and temperature remained on-5 ℃ 1 hour.Under-5 ℃, leach product, and use filtrate to wash solid, so that pure (2-{6-[((1R, 2R)-2-hydroxycyclopent base) amino to be provided] purine-9-yl } (4S, 5S, 3R)-and 5-[(2-fluorophenyl sulfo-) methyl]-Oxolan-3, the 4-glycol).

Similarly, step according to above 9A (preparation 1 or prepare 2), but with 2-{6-[(2-hydroxycyclopent base) amino] purine-9-yl } (4S, 5S, 3R)-5-(chloromethyl) Oxolan-3, other isomer of 4-glycol replaces 2-{6-[((1R, 2R)-and 2-hydroxycyclopent base) amino] purine-9-yl } (4S, 5S, 3R)-5-(chloromethyl) Oxolan-3, the 4-glycol makes following compound:

C. The preparation of the compound of Formula I wherein changes R

Similarly, according to the step of above 9A (preparation 1 or prepare 2), but with other thiophenol replacement 2-fluoro thiophenol of chemical formula RSH, make other compound of Formula I.

Embodiment 10

In conjunction with mensuration-DDT ₁Cell

Cell cultures

At 95% air and 5%CO ₂Damping atmosphere in, utilize the improved Eagle medium of Du Baike (Dulbecco) (DMEM), make DDT cell (hamster vas deferens smooth muscle cell system) be grown to individual layer in culture dish, wherein the improved Eagle medium of Dulbecco (DMEM) contains the amphotericin B of 2.5 μ g/ml, the penicillin G of 100U/ml, the Vetstrep of 0.1mg/ml and 5% foetal calf serum.Cell is gone down to posterity weekly cultivate twice by being dispersed in.Then with cell inoculation in growth medium, density is 1.2 * 10 ⁵Individual cell/plate, then pre-4 days of merging experimentized later at about 1 day.

Membrane product

(2 * 10mL) cells twice that adhere to of washing are being cleaned plate by means of rubber policeman (Rubber end glass paddle, rubber policeman) under 4 ℃ in the 50mM of 5ml Tris-HCl damping fluid (pH7.4), then 10 seconds of homogenizing suspension with HBSS.Centrifugal suspension 10 minutes under 27,000 * g then.As mentioned above, by rotation with centrifugal piller is resuspended in the homogenizing damping fluid.Final piller is resuspended in the Tris-HCl damping fluid of 1 volume 50mM, and (pH7.4 contains the MgCl of 5mM ₂) in, be used for A ₁AdoR measures.For [ ³⁵S] GTP γ S is in conjunction with mensuration, final piller is resuspended among the Tris-HCl of 50mM, its pH7.4, contains the MgCl of 5mM ₂, the NaCl of 100mM and the dithiothreitol (DTT) of 1mM.Then this film suspension was placed liquid nitrogen 10 minutes, melt (thawing) and be used for then measuring.By Bradford ^TMMeasure test kit and utilize bovine serum albumin protein content to be measured as standard.

Competitive binding assay

By (volume of 5 * liver mass, pH=7.4) homogenate prepares porcine striatal in the Tris of 50mM damping fluid.At 4 ℃ and 19, under the 000rpm after centrifugal 25 minutes, abandoning supernatant, and repeat twice of this process.The compound of Formula I is measured to determine that they are to porcine striatal membrane product or DDT ₁A in the membrane product ₁The avidity of acceptor.Briefly, handle porcine striatal film or the DDT of 0.2mg with adenosine deaminase and 50mM Tris damping fluid (pH=7.4) ₁Cytolemma then mixes.The DMSO storing solution of the serial dilution of the The compounds of this invention of adding 2 μ L in the film of pig, its concentration is that 100 micromoles are to 10nM.Contrast (sample) receives only the DMSO of 2 microlitres, be added in then the antagonist that is used for porcine striatal in the Tris damping fluid (50mM, pH are 7.4) [ ³H] 8-cyclopentyl xanthine (CPX) or be used for DDT ₁The agonist of film [ ³H] 2-chloro-6-UK 80882 (CCPA), to obtain the ultimate density of 2nM., utilize the film collector and utilize the repeatedly washing (3 *) of film to come filtering solution after 2 hours at 23 ℃ of following incubations.In the flicker mixture that tritiate CPX replacement amount is provided (scintillation cocktail) or the competition of the compound by Formula I in conjunction with counting filter disc (filter disk).

This mensuration shows that the compound of Formula I is for A ₁Adenosine Receptors demonstrates higher, medium or lower avidity.

Embodiment 11

[ ³⁵S] GTP γ S is in conjunction with mensuration

A ₁The agonist stimulation [ ³⁵S] GTP γ S is in conjunction with being by being measured by improving one's methods of people (1993) institute described method such as people such as Giersckik (1991) and Lorenzen.Under 30 ℃, incubation membrane protein in the volume of 0.1ml (30～50 μ g) 90 minutes wherein contains the Tris-HCL damping fluid (pH7.4) of 50mM, the MgCl of 5mM in the volume of 0.1ml ₂, 100mM dithiothreitol (DTT), the adenosine deaminase of 0.2 unit/ml, 0.5%BSA, 1mM EDTA, 10mM GDP, the 0.3nM[of NaCl, 1mM ³⁵S] GTP γ S and the CPA that contains or do not contain different concns.Non-specific binding is to measure by the GTP γ S that adds 10 μ M.The combination that agonist stimulates is determined as the difference between the basis combination that the total binding under the situation of CPA is arranged and do not having to measure under the situation of CPA.Previous report shows, agonist stimulate [ ³⁵S] GTP γ S is in conjunction with depending on (Gierschik et al., 1991 of existing of GDP; Lorenzen et al., 1993; Traynor﹠amp; Nahorski, 1995).In preliminary experiment, discover, 10 μ MGDP provide the CPA dependency [ ³⁵S] GTP γ S bonded optimal stimulus, thereby with this concentration be used for all research.In saturation experiments, with the GTP γ S incubation 0.5nM of 0.5～1000nM [ ³⁵S] GTP γ S.When incubation finishes, as mentioned above, filter every kind of suspension and measure the radioactivity that keeps.

This mensuration shows that the compound of Formula I is A ₁The partially or completely agonist of Adenosine Receptors.

Embodiment 12

CAMP measures

The scintillation proximity assay (SPA) that utilizes rabbit antibody to carry out points to cAMP, wherein utilizes adenosine 3 ', 5 '-cyclic phosphoric acid 2 '-O-succinyl-3-[ ¹²⁵I] the additional tracer of iodotyrosine methyl esters and the fluorescent microsphere (fluoromicrosphere) that contains anti-rabbit specific antibody, as described by Amersham Pharmacia Biotech (Biotrak cellular communication assays (Biotrak cell communication mensuration)).Briefly, at 37 ℃ of (5%CO ₂With 95% humidity) under, in the transparent 96 hole microtiter plates in bottom, cultivate DDT ₁Cell, wherein the concentration in opaque hole is 10 in the HBSS of 40 μ l ⁴To 10 ⁶Between the individual cells/well.At 37 ℃, under the situation of rolipram (50 μ M) and the existence of 5 μ M Forskolin, use DDT ₁Cell incubation under different concns partially or completely A of the present invention ₁Agonist (5 μ l) 10 minutes.Handle dissolved cell immediately by 10% bromination dodecyl TMA (TriMethylAmine), then utilize the microwell plate shaking table to shake with 5 μ l.After 5 minutes, in each hole, add immunoreagent solution (150 μ l contain isopyknic tracer, antiserum(antisera) and SPA fluorescence ball (fluorosphere)) at incubation (micropore) plate, then sealing (micropore) plate.At 23 ℃ down after 15～20 hours, by counting in the microtiter plate scintillometer measured in 2 minutes with the fluorescent microsphere bonded [ ¹²⁵I] amount of cAMP.Relatively be provided at the cAMP that the cell lysis exists later on the counting that utilizes similar program pin to the typical curve that cAMP produced.

This mensuration shows that the compound of Formula I is as A ₁Agonist has the partially or completely reduction of cAMP and is active on function.

Though the present invention is described according to its specific embodiment, it should be understood by those skilled in the art that under the situation that does not depart from true spirit of the present invention and scope, can carry out multiple improvement and be equal to replacement.In addition, can make multiple improvement so that specific situation, material, material composition, technology, a processing step or a plurality of processing step adapt to purpose of the present invention, spirit and scope.All such improvement include in the scope of appending claims.All patents cited above and publication are all incorporated herein with way of reference.

Claims

1. method that is used to prepare the compound of Formula I:

Wherein R is the phenyl of optional replacement;

Comprise:

Compound with chemical formula RSH under the situation that salt of wormwood exists and in inert solvent contacts with the compound of following chemical formula:

2. method according to claim 1, wherein, R is that 2-fluorophenyl, described 6-substituting group are 6-((1R, 2R)-2-hydroxycyclopent base amino), and described solvent is a N,N-dimethylacetamide.

3. method that is used to prepare the compound of following chemical formula:

Comprise:

Make the compound of following chemical formula:

Contact with the compound of following chemical formula:

4. method according to claim 3, wherein, described reaction is under the situation that alkali exists and carries out in inert solvent.

5. method according to claim 4, wherein, described alkali is selected from sodium hydroxide or triethylamine.

6. method according to claim 5, wherein, described alkali is triethylamine and described solvent is Virahol or methylene dichloride.

7. method according to claim 6, wherein, R is that 2-fluorophenyl and described 6-substituting group are 6-((1R, 2R)-2-hydroxycyclopent base amino).

8. method that is used to prepare the compound of following chemical formula:

Comprise:

Contact with the compound of following chemical formula with alkali:

9. method according to claim 8, wherein, described alkali is ammonia.

10. method according to claim 9, wherein, described reaction is carried out in methyl alcohol or methylene dichloride.

11. method that is used to prepare the compound of following chemical formula:

Comprise:

Under the situation that alkali exists, contact with the compound of following chemical formula with thionyl chloride:

12. method according to claim 11, wherein, described alkali is pyridine.

13. method according to claim 12, wherein, described reaction is carried out in methylene dichloride.

14. method that is used to prepare the compound of Formula I:

Wherein, R is the phenyl of optional replacement:

May further comprise the steps:

(b) use the compound of the product of alkali contact procedure (a) with generation chemical formula (13):

(c) under the situation that alkali exists with the product of 2-hydroxycyclopent base amine contact procedure (b) so that the compound of chemical formula (5) to be provided:

And

(d) compound and the step of usefulness chemical formula RSH under the situation that alkali exists

(c) product contact, wherein R as defined above.

15. method according to claim 14 wherein, is to carry out under the situation of inert solvent and alkali existence in reaction described in the step (a).

16. method according to claim 15, wherein, described inert solvent is a methylene dichloride and described alkali is triethylamine.

17. method according to claim 14 wherein, is an ammoniacal liquor at alkali described in the step (b).

18. method according to claim 17, wherein, described reaction is carried out in methyl alcohol or methylene dichloride.

19. method according to claim 14 wherein, is to carry out in inert solvent under the situation that triethylamine exists in reaction described in the step (c).

20. method according to claim 19, wherein, described inert solvent is Virahol or methylene dichloride.

21. method according to claim 19, wherein, R is that 2-fluorophenyl and described 6-substituting group are 6-((1R, 2R)-2-hydroxycyclopent base amino).

22. method according to claim 14 wherein, is to be selected from N in reaction described in the step (d), carries out in the solvent of dinethylformamide and N,N-dimethylacetamide.

23. method according to claim 22, wherein, described alkali is salt of wormwood.

24. method according to claim 22, wherein, described solvent is a N,N-dimethylacetamide.

25. method according to claim 22, wherein, R is that 2-fluorophenyl and described 6-substituting group are 6-((1R, 2R)-2-hydroxycyclopent base amino).