CN1032003C - Process for preparing pyrrolopyrimidine and intermediate - Google Patents

Abstract

The invention provides an improvement method for preparation of compounds or compound salts of novel general formula I (for the definitions of formula signs A, X, R1, R2, R3, R4, and n, see the description), the method comprises the method to make the compounds or compound salts of the general formula (II) (for the definitions of formula signs X, R1, R2, R3, R4, n, Y1, Y2, R8, and R9, see the description) occur the intra-molecular closed loop reaction to create the pyrrole and pyrimidine ring, if needed, the pyrrole ring can be reduced to generate the dihydro pyrrole ring. The compounds can be used as an anticancer agent.

Description

The method for preparing azolopyrimidines

The present invention relates to a kind of method for preparing new pyrrolo-[2,3-d] pyrimidine derivatives, this derivative can be used as anticarcinogen effectively; The invention still further relates to intermediate of this derivative and preparation method thereof.

In organism as formic acid, a carbon (C of formaldehyde etc. ₁) folic acid of unit transfer agent, in various enzymatic reaction systems, play a part coenzyme.For example, synthetic at biological nucleic acid, in amino acid peptide metabolism and the methane formation system.Specifically, in the biological nucleic acid synthesis system, this compound is absolutely necessary concerning the formylation reaction of two kinds of biosynthetic pathways of nucleic acid (being purine nucleotides approach and thymidine Nucleotide approach).In order to prove its biological activity, folic acid generally will be through two step reduction reactions, to be converted into active coenzyme.(methotrexate: be the medicine of known desmoenzyme consumingly (Tetrahydrofolate dehydrogenase) with similar compound MTX), this class medicine can be controlled its subordinate phase to methotrexate, becomes tetrahydrofolic acid (THFA) thereby suppressed dihydrofolate reduction.These medicines (play the effect of weakening DNA synthetic, cause necrocytosis) have been developed into cancer therapy drug, and have occupied very important position as clinical medicine at present.On the other hand, a kind of new tetrahydrochysene aminopterin anticarcinogen (5 has been proposed, 10-azatropylidene-5,6,7,8-tetrahydrochysene aminopterin: DDATHF), it has the basic framework of pteridine ring, but its mechanism of action is different with the medicine of this similar, and Tetrahydrofolate dehydrogenase is not suppressed active, and mainly be (the medical chemistry magazine 28,914 (1985)) finished by the mechanism that in the initial step of purine biosynthetic pathway, suppresses G-NH2 " nucleosides " transfer Formylase.

The MTX that has proposed clinical medicine commonly used mainly is the anticarcinogen by the folic acid antagonist effect one by one.But, therefore can not obtain satisfactory therapeutic effects because this medical instrument has stronger toxicity and little to the solid tumor effect.In addition, also exist the tumour cell resistance problem serious to this medicine.Therefore, the cancer therapy drug that can demonstrate improved efficient and cancer cells be demonstrated selective toxicity with the new mechanism of action is developed in special at present expectation.

According to above situation, the inventor has carried out research extensively and profoundly to the antitumour activity of the method for preparing no pteridine ring compound and this compound.So the inventor has proposed the novel method of pyrrolo-[2, the 3-d] pyrimidine derivatives of preparation 5-replacement, the selectivity that described derivative forms at productive rate, pyrrole ring etc. has the industrialization advantage; And the inventor finds that also a large amount of new compound that is made by this production method can have powerful inhibition growth activity to human body tumour cell.Just set up the present invention based on above discovery.

Therefore, the present invention relates to:

(1) a kind of method for preparing the compound or its salt that following general formula represents

Wherein encircle A represent one can be oxidized pyrrole ring; X represents amino, hydroxyl or sulfydryl; R ²And R ³Can be identical or different, represent respectively hydrogen or can substituted alkyl, alkenyl or alkynyl; R ⁴Represent OR ⁵(R wherein ⁵Represent hydrogen or can substituted alkyl) or NHCH (COOR ⁶) CH ₂CH ₂COOR ⁷(R wherein ⁶And R ⁷Represent respectively hydrogen or can substituted alkyl); N represents the integer of a 1-4, it is characterized in that said method comprising the steps of:

The compound or its salt that following general formula is represented

(X wherein, R ¹, R ², R ³, R ⁴Define as above with n; Y ¹And Y ²Represent oxygen or sulphur atom respectively; R ⁸And R ⁹Can be identical or different, representative can substituted alkyl respectively) in compound

During the base regeneration carbonyl or carry out ring-closure reaction afterwards, thereby form pyrrolo-[2,3-d] pyrimidine ring, need then, the pyrrole ring of ring  is reduced into the pyrroline ring, or/and with R ⁴And OR ⁵(R ⁵Definition is as above) be transformed into NHCH (COOR ⁶) CH ₂CH ₂COOR ⁷(R wherein ⁶And R ⁷Definition as above);

(2) compound (II) of explanation in the preceding paragraph (1);

(3) method introduced of a kind of preceding paragraph (1) for preparing the compound or its salt that following general formula represents:

Wherein X is amino, hydroxyl or sulfydryl; R ¹, R ²And R ³Can be identical or different, represent respectively hydrogen or can substituted alkyl, alkenyl or alkynyl; R ⁴Be OR ⁵(R ⁵Represent hydrogen or can substituted alkyl) or NHCH (COOR ⁶) CH ₂CH ₂COOR ⁷(R ⁶And R ⁷Represent respectively hydrogen or can substituted alkyl) and n be one 1～4 integer, it is characterized in that said method comprising the steps of: (i) compound or its salt of catalytic reduction general formula (II)

X wherein, R ¹, R ², R ³, R ⁴Define as above with n; R ⁸And R ⁹Can be identical or different, representative can substituted alkyl respectively; And Y ¹And Y ²Represent oxygen or sulphur atom respectively, described compound or its salt is contacted with acid or metal-salt, or (iii) reduce described compound or its salt with oxygenant; And

(4) method of the compound or its salt of a kind of preparation general formula (II) expression:

Wherein X represents amino, hydroxyl or sulfydryl; R ¹, R ²And R ³Can be identical or different, represent respectively hydrogen or can substituted alkyl, alkenyl or alkynyl; R ⁴Represent OR ⁵(R ⁵Represent hydrogen) or can substituted alkyl or NHCH (COOR ⁶) CH ₂CH ₂COOR ⁷(R ⁶And R ⁷Represent respectively hydrogen or can substituted alkyl), n represents one 1～4 integer, R ⁸And R ⁹Can be identical or different, representative can substituted alkyl respectively; Y ¹And Y ²Represent oxygen or sulphur atom respectively, it is characterized in that described method comprises to make the formula V compound

[wherein E represents CN, COOR ¹⁰, CSOR ¹⁰Or CSSR ¹⁰(R ¹⁰Representative can substituted alkyl), other symbol definition as above] with guanidine or its reactant salt.

The compound of above general formula (I) and (II) (wherein X is hydroxyl or sulfydryl) can exist with form with its tautomer equilibrium mixture.Below show the structural formula that the part of tautomerism easily takes place concerning X, and the balanced relationship between them.

For ease of expression, only introduced hydroxyl and sulfydryl form in the whole text in this specification sheets, and adopted corresponding systematic nomenclature.But, much less in both of these case, also comprise oxo or sulfo-isomer.

Though a large amount of unsymmetric molecules can be present in compound of the present invention (I) and (II) in, except by R ⁴Unsymmetrical carbon has outside the absolute configuration of S (L) in the side chain of the glutamic acid derivative of expression, and the absolute configuration of this class asymmetric center can also be the mixture of S-or R type or R-and S-type.In this case, have diastereomer, and the words that need can easily be separated by the routine separation to it with means of purification.These class methods of above-mentioned usefulness can include within the scope of the invention by isolating diastereomer.

With reference to following formula, X represents amino, hydroxyl or sulfydryl, and commonly used is amino; By R ¹, R ²And R ³Alkyl, the alkenyl or alkynyl of expression comprise C respectively _1-6Alkyl is as methyl, ethyl, propyl group and sec.-propyl; C _2-6Thiazolinyl is as vinyl, 1-methyl ethylene, 1-propenyl, allyl group and propadiene base; C _2-6Alkynyl, as ethynyl, 1-proyl and propargyl, thereby these are by R ²And R ³The group of expression can have nothing in common with each other in the repeating unit shown in the n.

R ¹, R ², and R ³Preferred example be hydrogen or the like.N represents one 1～4 integer, preferably represents 2 or 3.

By R ⁴The OR of expression ⁵R wherein ⁵And NHCH (COOR ⁶) CH ₂CH ₂COOR ⁷In R ⁶And R ⁷Hydrogen or can substituted alkyl respectively, for instance, the alkyl that can mention has C _1-5Low alkyl group is as methyl, ethyl, propyl group, sec.-propyl, butylidene, isobutyl-, sec-butyl, the tertiary butyl, n-pentyl, isopentyl, sec.-amyl sec-pentyl secondary amyl, neo-pentyl and tert-pentyl and benzyl or phenyl.R ⁴Preferred example has by formula OR ^5aOr-NHCH (COOR ^6a) CH ₂CH ₂COOR ^7a(R wherein ^5a, R ^6aAnd R ^7aRepresent for example C respectively _1-3Alkyl, weevil base and ethyl, or benzyl) group of expression, or the like.

Y ¹And Y ²Can be identical or different, represent oxygen or sulphur respectively, preferably oxygen.By R ⁶And R ⁹The alkyl of expression comprises C _1-5Low alkyl group, benzyl or phenyl (R ⁵, R ⁶And R ⁷In be described), that wherein commonly used is C _1-3Alkyl is as methyl or ethyl.

By above R ¹, R ²And R ³The expression alkyl, thiazolinyl and alkynyl and by R ⁵, R ⁶, R ⁷, R ⁸And R ⁹The alkyl of expression can have 1～3 substituting group.For instance, this class substituting group comprises halogen atom (as chlorine, fluorine, bromine and iodine), nitro, cyano group, about C _1-4Alkoxyl group (as methoxyl group, oxyethyl group, propoxy-, isopropoxy, n-butoxy, isobutoxy, sec-butoxy and tert.-butoxy), about C _1-4Alkyloyl (as formyl radical, ethanoyl, propionyl, positive butyryl radicals and isobutyryl), about C _1-4Alkanoyloxy (as methanoyl, acetoxyl group, propionyloxy, butyryl acyloxy and isobutyl acyloxy), about C _3-4Carbalkoxy (as methoxycarbonyl, ethoxycarbonyl, the positive third oxygen carbonyl, the different third oxygen carbonyl, positive butoxy carbonyl, isobutyl boc and tertbutyloxycarbonyl), trifluoromethyl, about C _1-4The about C of alkylthio (as methylthio group, ethylmercapto group, rosickyite base, iprotiazem base, positive butylthio, secondary butylthio and uncle's butylthio) _1-4Alkyl sulfinyl (as methyl sulfinyl, ethylsulfinyl-1 base, propyl group sulfinyl and butyl sulfinyl), about C _1-4Alkyl sulphonyl (as methylsulfonyl, ethylsulfonyl, third alkylsulfonyl and fourth alkylsulfonyl) or the like. work as R ⁵, R ⁶, R ⁷And R ⁸Or R ⁹When being benzyl or phenyl, it can be by C _1-4Alkyl (as methyl, ethyl, propyl group, sec.-propyl, butyl, isobutyl-, the sec-butyl and the tertiary butyl), C _1-3Thiazolinyl (as methylene radical, vinyl, 1-methyl ethylene, 1-propenyl, allyl group and propadiene base), C _2-3Alkynyl (as ethynyl, 1-proyl and propargyl) or the like replaces.

Below introduce in detail the method for preparation compound of the present invention (I) or its salt.

The reaction of carbonyl can by make compound (II) or its salt itself carry out decomposition reaction or a kind of suitable reaction solvent under carry out decomposition reaction and carry out, temperature of reaction is made an appointment with the boiling point (about below 150 ℃) of-40 ℃-described solvent, is preferably made an appointment with-10 ℃-75 ℃, lasted about 10 minutes-100 hours, preferably about 30 minutes-24 hours.For instance, the described decomposition reaction that can mention has hydrolysis reaction (method B-1) or the decomposition reaction (method B-2) under the acid anhydrous condition under catalytic reduction reaction (method A), the acidic conditions and makes the decomposition reaction of metal-salt that (method (1) or use the decomposition reaction (method C-2) of oxygenant particularly preferably is method B-1 or B-2.

Catalyst consumption is generally 1 mole of compound to be restored with about 0.005-2.0 mole, about 0.01-0.5 mole preferably among the method A.Can use platinum, rhodium Raney nickel etc. as catalyzer, add the acid (as acetate, trifluoroacetic acid, hydrochloric acid, sulfuric acid or the like) of trace simultaneously also to reacting favourable.The consumption of acid is generally 1 mole of about 0.01-100 mole of compound of wanting hydrolysis in method B-1.Preferably about 0.1-10 mole, and the example of acid comprises mineral acid, hydrochloric acid for example, Hydrogen bromide, sulfuric acid, nitric acid and phosphoric acid, organic acid, for example trifluoroacetic acid, trichoroacetic acid(TCA), methylsulfonic acid, Phenylsulfonic acid, tosic acid and camphorsulfonic acid, or the like, specifically, commonly used is mineral acid, example hydrochloric acid etc.

To be generally 1 mole of compound that will decompose be mineral acid with about 0.01-10 mole, preferably about 0.1-2 mole, and sour example to the consumption of acid among the method B-2, as hydrogenchloride, hydrogen bromide, mistake chloric acid, sulfuric acid, nitric acid, phosphoric acid or the like; Organic acid is as trifluoroacetic acid, trichoroacetic acid(TCA), methylsulfonic acid, Phenylsulfonic acid, tosic acid, camphorsulfonic acid or the like;

Lewi acid is as Zinc Chloride Anhydrous, Aluminum chloride anhydrous (AlCl ₃), anhydrous chlorides of rase is ferrous, titanium tetrachloride, tin tetrachloride, antimony chloride, cobalt chloride, cupric oxide, boron trifluoride-diethyl ether or the like.

The consumption of metal-salt is generally 1 mole of compound that will decompose among the method C-1 has cupric chloride, Silver Nitrate, silver suboxide, red precipitate, tellurium salt (as the nitric acid tellurium, trifluoroacetic acid tellurium or the like) with about 0.1-10 equivalent, preferably about 0.5-2 equivalent, and the example of metal-salt.

The consumption of chlorizating agent is generally about 0.25-10 equivalent, best 0.25-2 equivalent (to wanting the 1mol compound of oxidation) and the example of oxygenant among the method C-2 oxygen-twinkler, hydrogen peroxide, peroxybenzoic acid, metachloroperbenzoic acid, perchlorate (lithium perchlorate, cross silver chlorate, cross mercuric chlorate, cross chloric acid TBuA etc.), nitrosyl-sulfuric acid, alkyl nitrous acid (as amyl nitrite or the like), iodine, bromine, chlorine, N-bromine succinimide, SULPHURYL CHLORIDE, gansil or the like.

Concerning reaction solvent, the example that can be used for method A and B-1 has water, alcohol (as methyl alcohol, ethanol, Virahol, propyl alcohol, butanols, sec-butyl alcohol, isopropylcarbinol, ethylene glycol, first chloroethanol, ethoxy ethanol), acetic ester is (as methyl acetate, ethyl acetate), ether is (as methyl ether, ether, tetrahydrofuran (THF) ， diox, one glyme, diglyme), its hydrocarbon (benzene, toluene, dimethylbenzene), ketone (as, acetone), nitrile (as acetonitrile), pyridine, acid amides (as dimethyl formamide, N,N-DIMETHYLACETAMIDE), sulfoxide (as methyl-sulphoxide), tetramethylene sulfone and their solvent mixture; Under the situation of method A, preferably use methyl alcohol, ethanol, ethyl acetate, tetrahydrofuran (THF), diox or benzene; Under the situation of method B, a kind of water-containing organic solvent of normal use, wherein 0.01-100 restrains, preferably 0.1-10 restrains hydrophilic solvent [preferably pure (as methyl alcohol, ethanol etc.), acetic ester (as ethyl acetate), ester (as tetrahydrofuran (THF)) diox, etc.), ketone (as acetone etc.) or nitrile (as propionitrile etc.)] be included in 1 gram water.

The used amount of these solvents is generally: to whenever digesting compound (II) or its salt, with 1.0 to 2.000ml, preferred 5.0 to 100ml.

In method B-2, the available solvent has: acetic ester (as methyl acetate, ethyl acetate), ether (as, dme, ether, tetrahydrofuran (THF) ， diox, Monoethylene Glycol (MEG) dme, diglyme), aromatic hydrocarbon (as, benzene, toluene, dimethylbenzene), halohydrocarbon (as, methylene dichloride, chloroform, tetracol phenixin), ketone (as, acetone), nitrile (as, acetonitrile), Nitromethane 99Min., pyridine, dimethyl formamide or their suitable mixture; Preferred solvent be aromatic hydrocarbon (as, benzene, toluene etc.), halohydrocarbon (as, methylene dichloride, chloroform, tetracol phenixin etc.), or nitrile (as acetonitrile etc.).

Under application method C-1 and C-2 situation, the available solvent has: water, alcohol (as methyl alcohol, ethanol, propyl alcohol, Virahol, butanols, sec-butyl alcohol, the trimethyl carbinol, ethylene glycol, methoxyethanol, ethoxy ethanol), ether (as, dme, diethyl ether, tetrahydrofuran (THF), dioxane, Monoethylene Glycol (MEG) dme, diglyme), aromatic hydrocarbon (as, benzene, toluene, dimethylbenzene), halohydrocarbon (as, methylene dichloride, chloroform, tetracol phenixin etc.), acetone, acetonitrile and their suitable mixture.In method C-1 and C-2, preferred solvent is a methyl alcohol, ethanol, the mixture of tetrahydrofuran (THF) ， diox or acetonitrile and these solvents and water.As for for the regeneration carbonyl (＞=CO) should use the sort of method can be according to-Y '-R ⁸With-Y ²-R ⁹Chemical property etc. decide, usually, method B-1 is commonly used for preferred method.

At the carbonyl regeneration period and afterwards, in the intramolecular ring-closing reaction in the step of preparation The compounds of this invention (I) or its salt, carbonyl usually with amino autocondensation on pyrimidine ring, thereby generate pyrrolo-[2,3-d] pyrimidine ring.

Specifically, in method B-1 and B-2, the regeneration of carbonyl and intramolecular ring-closing reaction carry out very fast, and the product productive rate that obtains is better.At method A, among C-1 and the C-2, what the existence of acid catalyst can promote ring-closure reaction carries out and has improved productive rate fast.As acid catalyst, that can mention has: the mineral acid that describes in detail in method B-1 and B-2, organic acid or Lewis acid.

If desired, the ring  that obtains thus be pyrrole ring compound (I-1) can by catalytic reduction be easy to change into the ring  be the compound (I-2) of pyrroline ring.This catalytic reduction reaction is comparatively favourable with above-mentioned method A.

Compound (I-1) or (I-2) or their salt, wherein R ⁴Be OR ⁵, R ⁵For can substituted hydrocarbon, also can carry out said ester and decompose or hydrolysis reaction, thereby obtain R ⁵Be the compound of hydrogen, change this compound into R subsequently ⁴Be NHCH (COOR ⁶) CH ₂CH ₂COOR ⁷Compound.Ester decomposition reaction and change NHCH (COOR into ⁶) CH ₂CH ₂COOR ⁷Reaction can carry out [J.F.W.MCOmine, " Protective Groups in OrganieChemistry ", Plenum Press, Lonolon and New York (1973) by currently known methods; And M.Fieoer andL.Fieser, " Reagent for Orgamc Synthcrio ", Volo 1 to 13, Wiley-Interscience, NewYork, London, Sydney and Toronto (1969-1988)].

In addition, at compound (I) with (II) or in their salt, amino by the X representative, hydroxyl or sulfydryl can change into other group [a Separate Volume ofTanpakushitsu Kakusan Kohso (Protein Nuclcic Acids and EnEymes) by the conversion of the substituting group conversion reaction on the known pyrrole ring in the document, " ChemicalSynthcsis of Nucleic Acids ", Kyoritsu Publishing Co. (1968)].

The compounds of this invention (I) or its salt by the above-mentioned steps preparation can pass through common method, as concentrated, and solvent extraction, chromatogram, recrystallizations etc. are separated from reaction mixture or purifying.

Can prepare by the reactions steps shown in following as the The compounds of this invention (II) of intermediate or its salt in the methods of the invention.

In the above in the formula, Y ¹, Y ², R ¹, R ², R ³, R ⁴, R ⁸, R ⁹As above define with n, E represents CN group or COOR ¹⁰, CSOR ¹⁰Or CSSR ¹⁰(R wherein ¹⁰For can substituted alkyl, and Z be a halogen atom, as chlorine, and bromine, iodine).COOR ¹⁰, CSOR ¹⁰Or CSSR ¹⁰In R ¹⁰Can be as to R ⁵To R ⁹That describes in detail can substituted alkyl, and preferred group is C _1-4Alkyl, as methyl and ethyl etc., or benzyl etc.Above-mentioned reactions steps is as detailed below:

Steps A

This step comprise with

Be added to two key (R of compound (III) ⁸Thereby-Y '-C=CH-) goes up the step that generates compound (IV).

The consumption of relativization compound (III) is generally 0.5 to 4 molar equivalent, preferred 0.8 to 1.5 molar equivalent.This is reflected at suitable solvent and exists down, in temperature of reaction be-10 ℃ to used reaction solvent boiling points (height is to 150 ℃), carried out 30 minutes to 48 hours under preferred 0 ℃ to 100 ℃.The solvent that uses in the reaction has: alcohol (as, methyl alcohol, ethanol), ether (as, dme, diethyl ether, tetrahydrofuran (THF) ， diox, the Monoethylene Glycol (MEG) dme, diglyme, nitrile (as acetonitrile), ester (as ethyl acetate), halohydrocarbon (as, methylene dichloride, chloroform, tetracol phenixin), aromatic hydrocarbon is (as benzene, toluene, dimethylbenzene) or their suitable mixture. in carrying out this reaction, exposure or add organo-peroxide and also can make this reaction carry out more favourablely.Said organo-peroxide comprises: tert-butyl hydroperoxide, Peracetic Acid, benzoyl hydroperoxide, m-chloroperoxybenzoic acid etc.The compound (IV) that is obtained by this step has relative reactive behavior, and can separate by above-mentioned ordinary method in this step, also can separate to be directly used in next step reaction.

Step B:

In the presence of suitable solvent, be-10 ℃ of boiling points (up to 100 ℃) in temperature of reaction to reaction solvent, the compound (IV) and the R that steps A are obtained under preferred 0 ℃ to 50 ℃ ⁹-Y ²Alcohol that-H represents or thiol reactant 10 minutes to 24 hours, thus change compound (V) into.The solvent that uses in this reaction has: ether (as dme, diethyl ether, tetrahydrofuran (THF) diox, Monoethylene Glycol (MEG) dme, diglyme), nitrile (as acetonitrile), ester (as ethyl acetate), and halohydrocarbon (as methylene dichloride, chloroform, tetracol phenixin), aromatic hydrocarbon (as benzene, toluene, dimethylbenzene etc.) or their suitable mixture.In addition, by R ⁹-Y ²Can be excessive when the alcohol of-H representative or its mercaptan use as solvent.The compound that obtains thus (V) can be separated from reaction mixture by above-mentioned ordinary method, also can separate.This reaction mixture is directly used in next step reaction as raw material.

Step C

In suitable solvent, with guanidine or its salt (as with the salt that forms with acid described in the top method B-1 etc.) handle compound (V), the cyano group of guanidine and compound (V), ester group or sulfo-ester group react and cause that simultaneously cyclization forms pyrimidine ring, generates The compounds of this invention (II) or its salt thus.This is reflected at 0 ℃ to 150 ℃, and preferred 20 ℃ to 100 ℃ were carried out 1 to 48 hour.This be reflected at carry out under the alkaline condition more favourable.Used alkali comprises metal alkoxide, as sodium methylate, and sodium ethylate, potassium tert.-butoxide etc.The available reaction solvent has: methyl alcohol, ethanol, propyl alcohol, the trimethyl carbinol, dimethyl sulfoxide (DMSO), hexamethyl phosphonic amide or their suitable mixture etc.

Compound of the present invention (II) and the available conventional isolated or purified method of the compound as initiator or intermediate (III) to (V) that in these steps, prepares, as concentrating solvent extraction, chromatogram, recrystallizations etc. obtain isolated or purified from the respective reaction mixture.

That make by preparation method of the present invention or compound (I) and compound (II) or its intermediate can salt form exist; preferred pharmaceutical salts, the available basic salt has: with basic metal, alkaline-earth metal; no noxious metals; the salt that the ammonium of ammonium and replacement forms, as with sodium, lithium; cobalt, magnesium, aluminium, zinc, ammonium, trimethyl ammonium, triethyl ammonium, triethanol ammonium; pyridine, or the salt that forms such as the pyridine that replaces by for example carbamyl or halogen (as chlorine, bromine).Acid-salt has: with the salt that mineral acid forms, example hydrochloric acid, sulfuric acid, nitric acid, and phosphoric acid and boric acid and the salt that forms with organic acid (as oxalic acid, tartrate, acetate, trifluoroacetic acid, methylsulfonic acid, Phenylsulfonic acid is to a toluenesulphonic acids and camphorsulfonic acid etc.).

Compound (I) or their salt pair mouse tumor cell strain (P388, L1210, L5178Y, the B16 melanoma, MethA, Louis's tumour, the S180 sarcoma, intestinal cancer and Colon38) and human body tumour cell strain (HL60 and KB) demonstrate fabulous anti-tumor activity, meanwhile, they to the homoiothermous tumour of attack (as melanoma, sarcoma, mastocytoma, cancer, tumour formation etc.) have an effect that reduces tumor promotion and can prolong the warm-blooded animal life-span of suffering from tumour.In addition, the pharmaceutical preparation that contains compound (I) or their salt can be used as safe antineoplastic agent and is used for the treatment of the warm-blooded animal that suffers from tumour, especially mammal (as mouse, rat, cat, dog, rabbit etc.).

As under the antineoplastic agent situation, compound (I) or its salt can be according to a conventional method by with pharmaceutical carrier, vehicle, thinner etc. are mixed and made into by the oral or various dosage forms of parenterai administration, as pulvis, stick, tablet, capsule, suppository and injection liquid.Though dosage will be according to the kind of treatment animal, the kind of disease, symptom, type of compounds, variations such as route of administration, but under oral situation, compound (I) or its salt can be by per daily dose 2.0-200mg/kg body weight, preferred 5.0-100mg/kg body weight, deliver medicine to above-mentioned warm-blooded animal, or under the parenterai administration situation, press per daily dose 1.0-100mg/kg body weight, preferred 2.0-50mg/kg body weight delivers medicine to above-mentioned warm-blooded animal.The injection liquid medication comprises intramuscular, intraperitoneal, subcutaneous and intravenous injection etc.

The preparation of above-mentioned various formulations can be undertaken by following currently known methods.Oral preparations, as tablet, can by add an amount of tackiness agent (as, hydroxypropylcellulose, HPMC, tight gel etc.), disintegrating agent (as, starch, calcium carboxymethylcellulose etc.), lubricant (as, Magnesium Stearate, talcum etc.) wait and prepare.

Parenteral formulation, as injection liquid, can by add an amount of tonic (as, glucose, D-Sorbitol Powder, D-N.F,USP MANNITOL, sodium-chlor etc.), sanitas (as, benzylalcohol, chlorobutanol, to an oxo methyl benzoate, to oxo propyl benzoate etc.), damping fluid (as phosphoric acid buffer, sodium acetate buffer etc.) waits and prepares.

The object lesson of preparation tablet is: with every is benchmark; with 1.0-25mg The compounds of this invention (I) or its salt; the 100-500mg lactose; 50-100mg W-Gum and 5-20mg hydroxypropylcellulose mix by ordinary method; with this mixture granulation; mix with W-Gum and Magnesium Stearate subsequently, be pressed into diameter 3-10mm, the tablet of heavy 100-500mg.This tablet that makes can obtain the enteric coating tablet thus by coming dressing with the Viscotrol C (0.5-2mg) that contains the acetone-alcohol mixture of Vltra tears (about 10-20mg) and concentration 5-10% (this amount be benchmark with every).

The specific examples of preparation injection liquid is, amount with each ampoule is a benchmark, the solution of the 2ml isotonic saline solution of 2.0-50mg The compounds of this invention (I) sodium salt is loaded in the ampoule, then it is sealed and 110 ℃ of heat sterilizations 30 minutes, or the described sodium salt of 2.0-50mg is dissolved in the solution of 2ml sterile distilled water of 10-40mg N.F,USP MANNITOL or sorbyl alcohol, the ampoule of then solution that obtains being packed into is subsequently with this ampoule lyophilize, seal then, obtain injection liquid.When using, frozen preparation sealed opens, make that compound concentration is 1-25mg/ml in the solution thereby active ingredient and physiological saline mixed, obtain thus injection liquid can be used for subcutaneous, vein or intramuscular administration.

In sum, preparation method of the present invention is initial with inexpensive industrial coarse raw materials, and provides as medicinal anti-tumor agent comprising salmosin with the reactions steps of shortening and the reaction yield of raising.In addition, from response capacity and efficient, this method is easily simple especially, and from preparing easy viewpoint, this method is more desirable, therefore, the invention provides the also industrial favorable method of [2,3-d] pyrimidine derivatives of preparation 5-substituted azole.

Following mask body is introduced of the present invention with reference to example and example.

Reference example 1

The preparation of 4-(4-methoxyl group-3-butenyl)-t-butyl perbenzoate

In 0 ℃, the tetrahydrofuran solution (11.0ml) of 1.0 moles of potassium tert.-butoxides is added in the toluene solution (12ml) of chlorination (methoxyl methyl) triphenyl phosphonium (3.77g), stir after 10 minutes, under same temperature, the toluene solution (10ml) of 4-(4-propoxy-) t-butyl perbenzoate (2.34g) is added drop-wise in this mixture, stirred 20 minutes in 0 ℃ subsequently.This reaction solution is mixed with ether (40ml), tell organic layer, water and saturated sodium-chloride water solution wash successively then, use anhydrous sodium sulfate drying, remove solvent under reduced pressure, and the resistates that obtains is handled with hexane, filter out triphenylphosphine oxidation thing then.Concentrating under reduced pressure filtrate, resistates obtains title compound (1.92g) by column chromatography (80g silica gel, ether-hexane=20: 1) purifying.

IR (pure): 2980,2945,1715,1655,1610,850cm ^-1

¹H-NMR(CDCl ₃)：1.59(9H，s，)，2.24(1.2H，td，J＝8Hz，7Hz)，2.39(0.8H，td，J＝8Hz，7Hz)，3.48(1.8H，s)，3.56(1.2H，s)，4.33(0.4H，td，J＝7Hz，6Hz)，4.71(0.6H，dt，J＝13Hz，7Hz)，5.88(0.4H，d，J＝6Hz)，6.28(0.6H，d，J＝13Hz)，7.21(2H，d，J＝8Hz)，7.91(2H，d，J＝8Hz).

Reference example 2

4-[4,4-dicyano-3-(dimethoxy methyl) butyl] preparation of t-butyl perbenzoate

Under argon atmospher, the compound (1.91g) that bromine propane dinitrile (1.27g) and reference example 1 are obtained is dissolved in the methylene dichloride (66ml), add molecular sieve (3A, 1.0g) after, reaction mixture with analysis not with cover with the ultraviolet radiation of ultraviolet lamp 2 hours.(4ml) is added in the reaction mixture with methyl alcohol, stirs 10 minutes, and impouring contains in the frozen water of 2N wet chemical (5ml) then.Then use dichloromethane extraction.Organic layer washes with water, uses anhydrous sodium sulfate drying.Remove solvent under reduced pressure, the resistates that obtains obtains being the title compound (2.08g) of colorless oil with column chromatography (75g silica gel, ethyl acetate-hexane=1: 10) purifying.

IR (pure): 2980,2945,2840,2250,1710,1606,845cm ^-1

¹H-NMR(CDCl ₃)：1.60(9H，s)，1.90-2.20(2H，m)，2.20-2.32(1H，m)，2.89(2H，t，J＝8Hz)，3.39(3H，s)，3.46(3H，s)，4.13(IH，d，J＝4Hz)，4.36(1H，d，J＝5Hz)，7.23(2H，d，J＝8Hz)，7.95(2H，d，J＝8Hz).

Reference example 3

The preparation of 4-(5-methoxyl group-4-pentenyl)-t-butyl perbenzoate

Press and reference example 1 described same step, 4-(4-butoxy) t-butyl perbenzoate (993mg) is handled with chlorination (methoxyl methyl) triphenyl phosphonium, obtain being the title compound (918mg) of colorless oil.

IR (pure): 2980,2940,2860,1710,1660,1603,860,845cm ^-1

¹H-NMR(CDCl ₃)：1.55-1.76(2H，m)，1.59(9H，s)，1.96(0.6H，dt，J＝7Hz，7Hz)，2.10(0.4H，tdd，J＝7Hz，7Hz，7Hz)，2.66(2H，t，J＝8Hz)，3.51(1.8H，s)，3.59(1.2H，s)，4.35(0.4H，td，J＝7Hz，6Hz)，4.73(0.6H，dt，J＝13Hz，7Hz)，5.91(0.4H，dt，J＝6Hz，1Hz)，6.29(0.6H，d，J＝13Hz)，7.21(2H，d，J＝8Hz)，7.89(0.8H，d，J＝8hz)，7.90(1.2H，d，J＝8Hz).

Reference example 4

4-[5,5-dicyano-4-(dimethoxy methyl) amyl group] preparation of t-butyl perbenzoate

By reference example 2 described same steps,, obtain being the title compound (202mg) of colorless oil with reference to the compound (276mg) and the reaction of bromine propane dinitrile that example 3 obtains.

IR (pure): 2975,2930,2245,1710,1605,860,845cm ^-1

¹H-NMR(CDCl ₃)：1.59(9H，s)，1.60-1.92(4H，m)，2.20-2.30(1H，m)，2.73(2H，t，J＝7Hz)，3.40(3H，s)，3.45(3H，s)，4.11(1H，d，J＝4Hz)，4.31(1H，d，J＝5Hz)，7.24(2H，d，J＝8Hz)，7.93(2H，d，J＝8Hz).

Reference example 5

The preparation of 4-(6-methoxyl group-5-hexenyl) t-butyl perbenzoate

By reference example 1 described same step, handle 4-(5-butoxy) t-butyl perbenzoate (476mg) with chlorination (methoxyl methyl) triphenyl phosphonium, obtain being the title compound (430mg) of colorless oil.

IR(neat)：2940，1715，1650，1605，1455，850cm ^-1

¹H-NMR(CDCl ₃)：1.32-1.44(2H，m)，1.59(9H，s)，1.52-1.70(2H，m)，1.94(1.2H，td，J＝8Hz，7Hz)，2.09(0.8H，td，J＝8Hz，7Hz)，2.65(2H，t，J＝8Hz)，3.49(1.8H，s)，3.58(1.2H，s)，4.31(0.4H，td，J＝7Hz，6Hz)，4.70(0.6H，dt，J＝13Hz，7Hz)，5.88(0.4H，d，J＝6Hz)，6.28(0.6H，d，J＝13Hz)，7.21(2H，d，J＝8Hz)，7.90(2H，d，J＝8Hz).

Reference example 6

4-[6,6-dicyano-5-(dimethoxy methyl) hexyl] preparation of t-butyl perbenzoate

By the same step of reference example 2,, obtain being the title compound (432mg) of colorless oil with reference to the compound (420mg) and the reaction of bromine propane dinitrile that example 5 obtains.

IR(Neat)：2940，2250，1715，1610，1455，845cm ^-1

¹H-NMR(CDCl ₃)：1.48-1.81(6H，m)，1.59(9H，s)，2.18-2.28(1H，m)，2.71(2H，t，J＝7Hz)，3.40(3H，s)，3.46(3H，s)，4.10(1H，d，J＝4Hz)，4.31(1H，d，J＝5Hz)，7.23(2H，d，J＝8Hz)，7.92(2H，d，J＝8hz).

Example 1

4-[3-(2,4,6-triamino-pyrimidine-5-yl)-4,4-dimethoxy butyl] preparation of t-butyl perbenzoate

Under argon atmospher, the trimethyl carbinol suspension (30ml) of guanidinesalt hydrochlorate (640mg) is mixed with the tetrahydrofuran solution (6.70ml) of 10 moles of potassium tert.-butoxides, stirred subsequently 10 minutes, t-butanol solution (10ml) with reference to the compound (2.00g) of example 2 is added in this mixture, and postheating refluxed 2 hours.The reaction soln impouring is contained in the water (200ml) of 1.0N aqueous potassium hydrogen sulfate (1ml), then use dichloromethane extraction.The organic layer anhydrous sodium sulfate drying, solvent evaporated under reduced pressure.Resistates column chromatography (50g silica gel; Methylene chloride-methanol=30: 1 → 15: 1) purifying obtains being the title compound (2.17g) of colourless amorphous.

IR(KBr)：3475，3360，3200，2975，2930，1710，1607，1563，1430，843，800cm ^-1

¹H-NMR(CDCl ₃)：1.58(9H，s)，1.86-2.05(1H，m)，2.25-2.53(2H，m)，2.57-2.80(2H，m)，3.45(8H，s)，3.48(3H，s)，4.35(1H，d，J＝3Hz)，4.36(2H，brs)，4.48(2H，brs)，5.21(2H，brs)，7.18(2H，d，J＝8Hz)，7.88(2H，d，J＝8Hz).

Example 2

N-[4-[2-(2,4-diamino-7H-pyrrolo-[2,3-d]-pyrimidine-5-yl) ethyl] benzoyl]-preparation of C-glutamate diethyl ester

The compound (200mg) that example 1 is obtained is dissolved in trifluoroacetic acid (1ml) and the water (20mg), then stirring at room 2 hours.Remove trifluoroacetic acid under reduced pressure, be suspended in the dimethyl formamide (2ml) in resistates and the L-diethyl glutamate hydrochloride (172mg) that 70 ℃ of vacuum-dryings obtain then.In 0 ℃ diethyl phosphorocyanidate (diethylphxphorocyanidate) dimethyl formamide solution (2ml) (82mg) is added in this suspension, then stirred 15 minutes, under same temperature, the dimethyl formamide solution of triethylamine (218mg) is added drop-wise in this mixture again, then stirred 30 minutes in 0 ℃, again in stirring at room 2 hours.Remove solvent under reduced pressure, the resistates that obtains obtains being the title compound (195mg) of colourless amorphous with column chromatography (15g silica gel, the methylene dichloride of telling from strong aqua, the methylene dichloride of telling from the ethanol of 10% ammoniacal liquor=40: 1 → 30: 1) purifying.

IR(KBr)：3375，3200，2980，2930，1735，1640，1605，1572cm ^-1

¹H-NMR(CDCl ₃)：1.23(3H，t，J＝7Hz)，1.31(3H，t，J＝7Hz)，2.10-2.40(2H，m)，2.48(2H，dd，J＝6Hz，6Hz)，3.00(4H，brs)，4.12(2H，q，J＝7Hz)，4.25(2H，q，J＝7Hz)，4.61(2H，brs)，4.75-4.86(1H，m)，4.95(2H，brs)，6.40(1H，s)，7.13(1H，d，J＝7Hz)，7.22(2H，d，J＝8Hz)，7.74(2H，d，J＝8Hz)，8.55(1H，brs).

Example 3

N-[4-[2-(2,4-diamino-7H-pyrrolo-[2,3-d] pyrimidine-5-yl) ethyl] benzoyl]-preparation of L-L-glutamic acid

Tetrahydrofuran (THF) one water mixed liquid of the compound (80mg) that obtains through example 2 (2: 1, add 1.0N aqueous sodium hydroxide solution (0.497ml) in 3ml), then in stirring at room 1 hour.Concentrating under reduced pressure reaction solution to cumulative volume is 1ml, filter out the insolubles that obtains by the Milliporl strainer after, solution is cooled to 0 ℃ and mix with acetate (0.1ml).Filter and collect the crystallization that obtains, use the frozen water thorough washing, in 70 ℃ of drying under reduced pressure, crystalline title compound (61mg) obtains being white in color.

IR(KBr)：3320，1660，1637，1540cm ^-1

¹H-NMR(Me ₂SO-d ₆)：1.85-2.20(2H，m)，2.46(2H，t，J＝8Hz)，2.96(4H，brs)，4.30-4.45(1H，m)，5.49(2H，brs)，6.13(2H，s)，6.37(1H，s)，7.33(2H，d，J＝8Hz)，7.80(2H，d，J＝8Hz)，8.46(1H，d，J＝7hz)，10.34(1H，brs).

Example 4

4-[4-(triaminopyrimidine-5-yl)-5,5-dimethoxy amyl group] preparation of t-butyl perbenzoate.

Press the same step described in the example 1, with reference to compound (190mg) and the reaction of guanidinesalt hydrochlorate that example 4 obtains, powdered title compound (214mg) obtains being white in color.

IR(KBr)：3480，3380，3200，2980，2940，1715，1610，1570，1440，850，805cm ^-1

¹H-NMR(CDCl ₃)：1.40-1.65(3H，m)，1.59(9H，s)，1.75-2.05(1H，m)，2.62(2H，t，J＝7Hz)，2.81(1H，ddd，J＝11Hz，3Hz，1Hz)，3.46(3H，s)，3.50(3H，s)，4.36(1H，d，J＝7Hz)，4.49(4H，brs)，5.16(2H，brs)，7.18(2H，d，J＝8Hz)，7.88(2H，d，J＝8Hz).

Example 5

4-[3-(2,4-diamino-7H-pyrrolo-[2,3-d] pyrimidine-5-yl) propyl group] preparation of t-butyl perbenzoate

The compound (206mg) that example 4 is obtained be dissolved in tetrahydrofuran (THF)-water mixed liquid (3: 1,8ml) in, 1N hydrochloric acid (4.8ml) is added in this solution, then in stirring at room 18 hours.Reaction solution is then used dichloromethane extraction with 1N sodium hydroxide solution (4.8ml) neutralization, and the extract layer anhydrous sodium sulfate drying removes solvent under reduced pressure.Obtain resistates column chromatography (10g silica gel, methylene chloride-methanol=15: 1) purifying, crystalline title compound (126mg) obtains being white in color.

m.p.172-173℃

IR(KBr)：3335，3180，2975，2935，1710，1607，1287，1163，1110cm ^-1

¹H-NMR(Me ₂SO-d ₆)：1.54(9H，s)，1.77-1.90(2H，m)，2.68(2H，t，J＝8Hz)，2.72(2H，t，J＝8Hz)，5.54(2H，brs)，6.11(2H，brs)，6.45(1H，s)，7.33(2H，d，J＝9Hz)，7.82(2H，d，J＝8Hz)，10.51(1H，s).

Embodiment 6

N-[4-[3-(2,4-diamino 7H-pyrrolo-[2-3-d] pyrimidine-5-yl) propyl group] benzoyl]-preparation of L-glutamate diethyl ester

(A) compound that example 5 is obtained (381mg) is dissolved in the trifluoroacetic acid (3ml), this solution was in stirring at room 3 hours, remove trifluoroacetic acid under reduced pressure, in 70 ℃ of drying under reduced pressure, the resistates that obtains is suspended in dimethyl formamide (4ml) with diethyl glutamate hydrochloride (748mg) subsequently.(4ml) is added in this suspension in 0 ℃ of dimethyl formamide solution with diphenyl phosphoryl azide (858mg); stir subsequently; under same temperature; the dimethyl formamide solution (4ml) of triethylamine (631mg) is added drop-wise in this mixed solution; stirred 30 minutes in 0 ℃, then in room temperature restir 63 hours.Remove solvent under reduced pressure, the resistates that obtains obtains being the title compound (374mg) of colourless crystallization with column chromatography (15g silica gel, the dichloromethane-ethanol of the methylene dichloride of being told by strong aqua → tell from strong aqua=40: 1 → 30: 1) purifying.

(B) press example 2 described same steps, the compound that is obtained by example 4 can prepare this title compound.

¹H-NMR(Me ₂SO-d ₆)：1.17(3H，t，J＝7Hz)，1.20(3H，t，J＝7Hz)，1.80-2.20(4H，m)，2.44(2H，t，J＝7Hz)，2.68(2H，t，J＝7Hz)，2.72(2H，t，J＝7Hz)，4.05(2H，q，J＝7Hz)，4.11(2H，q，J＝7Hz)，4.35-4.50(1H，m)，5.34(2H，s)，5.91(2H，s)，6.42(1H，s)，7.31(2H，d，J＝8Hz)，7.80(2H，d，J＝8Hz)，8.66(1H，d，J＝8Hz)，10.51(1H，s).

Embodiment 7

N-[4-(3-(2,4-diamino-7H-pyrrolo-[2,3-d] pyrimidine-5-yl) propyl group] benzoyl]-preparation of L-L-glutamic acid

Press example 3 described same steps, the compound that example 6 the is obtained reaction that is hydrolyzed, crystalloid title compound (201mg) obtains being white in color.

m.p.220-221℃

IR(KBr)：3240，3200，2940，1660-1630，1540，1500，1397cm ^-1

¹H-NMR(Me ₂SO-d ₆)：1.75-2.20(4H，m)，2.35(2H，t，J＝7Hz(，2.68(2H，t，J＝7Hz)，2.71(2H，t，J＝7Hz)，4.30-4.47(1H，m)，5.53(2H，brs)，6.15(2H，s)，6.46(1H，s)，7.31(2H，d，J＝8Hz)，7.81(2H，d，J＝8Hz)，8.48(1H，d，J＝8Hz)，10.51(1H，s).

Example 8

4-[5-(triaminopyrimidine-5-yl)-6,6-dimethoxy hexyl] preparation of t-butyl perbenzoate

According to example 1 described same steps as, make compound (378mg) and Guanidinium hydrochloride reaction that reference example 6 obtained, obtain white crystalline title compound (433mg).

IR(KBr)：3475，3360，3220，2975，2930，1715，1640，1607，1563，1435，843，800cm ^-1

¹H-NMR(CDCl ₃)：1.14-1.32(2H，m)，1.45-1.72(3H，m)，1.58(9H，s)，1.86-2.04(1H，m)，2.56-2.68(2H，m)，2.72-2.83(1H，m)，3.47(3H，s)，3.52(3H，s)，4.39(1H，d，J＝8Hz)，4.36(2H，brs)，4.48(2H，brs)，5.21(3H，brs)，7.18(2H，d，J＝8Hz)，7.88(2H，d，J＝8Hz).

Embodiment 9

N-[4-[4-(2,4-diamino-7H-pyrrolo-[2,3-d] pyrimidine-5-yl) butyl] benzoyl]-preparation of L-glutamate diethyl ester

According to example 2 described same steps as, the compound (230mg) that is obtained with example 8 reacts, and obtains the title compound (228mg) of white powder.

IR(KBr)：3380，3200，2980，2930，1735，1640，1605，1572cm ^1-

¹H-NMR(CDCl ₃/CD ₃OD)：1.22(3H，t，J＝7Hz)，1.31(3H，t，J＝7Hz)，1.60-1.83(4H，m)，2.43-2.51(2H，m)，2.63-2.76(4H，m)，4.11(2H，q，J＝7Hz)，4.24(2H，q，J＝7Hz)，4.74-4.86(1H，m)，6.45(IH，s)，7.24(2H，d，J＝8Hz)，7.74(2H，d，J＝8Hz).

Example 10

N-[4-[4-(2,4-diamino-7H-pyrrolo-[2,3-d] pyrimidine-5-yl) butyl] benzoyl]-preparation of L-L-glutamic acid

According to example 3 described same steps as, the compound (103mg) that example 9 the is obtained reaction that is hydrolyzed obtains white crystalline title compound.

IR(KBr)：3340，3200，2930，1650，1635，1540cm ^-1

¹H-NMR(Me ₂SO-d ₆)：1.45-1.76(4H，m)，1.88-2.19(2H，m)，2.29-2.43(2H，m)，2.58-2.76(4H，m)，4.32-4.46(1H，m)，5.54(2H，brs)，6.16(2H，brs)，6.42(1H，s)，7.29(2H，d，J＝8Hz)，7.79(2H，d，J＝8hz)，8.52(1H，d，J＝7Hz)，10.48(1H，brs).

Example 11

N-[4-[3-(2,4-diamino-6,7-dihydro-5-pyrrolo-[2,3-d] pyrimidine-5-yl) propyl group] benzoyl]-preparation of L-glutamate diethyl ester

The compound (20mg) that example 9 is obtained is dissolved in 2% ethanol solution hydrochloride (20ml), and behind the adding platinum oxide (3mg), catalytic reduction reaction is 12 hours under hydrogen atmosphere.The elimination catalyzer, concentrated filtrate is to doing.With column chromatography (2.0g silica gel; Isolating methylene dichloride from dense ammonia: the resistates of purifying and obtaining isolating methylene dichloride from dense ammonia-ethanol=40: 1 → 30: 1) obtains title compound (4.8mg).

IR(KBr)：3350，2990，2945，1740，1610，1540，1508，1438cm ^-1

¹H-NMR(CDCl ₃)：1.23(6H，tx2，J＝7Hz)，1.43-1.80(3H，m)，1.85-2.77(7H，m)，2.95-3.30(2H，m)，3.58(1H，t，J＝11Hz)，4.07(2H，q，J＝7Hz)，4.20(2H，q，J＝7Hz)，4.25(1H，brs)，4.63-4.83(1H，m)，4.68(1H，brs)，7.00-7.23(1H，m)，7.13(2H，d，J＝8Hz)，7.67(2H，d，J＝8Hz).

Embodiment 12

N-[4-[3-(2,4-diamino-6,7-dihydro-5-pyrrolo-[2,3-d] pyrimidine-5-yl) propyl group] benzoyl]-preparation of L-L-glutamic acid

According to example 3 described same steps as, the compound (4.4mg) that example 11 the is obtained reaction that is hydrolyzed obtains title compound (3.2mg).

IR(KBr)：3700-2350，3215，1690-1620，1540cm ^-1

¹H-NMR(Me ₂SO-d ₆)：1.02-1.85(4H，m)，1.85-2.83(6H，m)，2.90-3.30(2H，m)，3.55(1H，t，J＝11Hz)，4.15-4.45(1H，m)，6.38(2H，brs)，6.77(2H，brs)，6.90(1H，brs)，7.22(2H，d，J＝8Hz)，7.74(2H，d，J＝8Hz)，8.22(1H，d，J＝7Hz).

Example 13

4-[4-(triaminopyrimidine-5-yl)-5, the preparation of 5-dimethoxy amylbenzene ethyl formate.

Under argon gas atmosphere, the solution (0.612ml) of 1.0 moles of sodium ethoxide in ethanol is joined in the alcohol suspension (1.0ml) of Guanidinium hydrochloride (58.5mg).In this solution, add 4-[5,5-dicyano-4-(dimethoxy-methyl) amyl group]-ethanolic soln (3.0ml) of ethyl benzoate (190mg) and reflux mixture 2 hours.(190mg) obtain by 4-(4-butoxy) ethyl benzoate by reference example 3 and 2.

Reaction mixture is poured in the water (10ml),, used the anhydrous sodium sulfate drying organic layer in dichloromethane extraction.Remove from extraction liquid with distillation under vacuum and to desolvate, resistates is with flash distillation column chromatography purifying (15g silica gel; Methylene dichloride: methyl alcohol=30: 1 → 15: 1), obtain title compound (214mg).

IR(KBr)：3460，3340，3180，2940，1710，1610，1565，1435，1275，1110，1060，805cm ^-1

¹H-NMR(CDCl ₃)：1.38(3H，t，J＝7.2Hz)，1.43-1.70(3H，m)，1.83-2.05(1H，m)，2.63(2H，t，J＝7Hz)，2.77-2.87(1H，m)，3.46(3H，s)，3.50(3H，s)，4.36(2H，q，J＝7.2Hz)，4.37(1H，d，J＝3.6Hz)，4.45(4H，brs)，5.10(2H，brs)，7.20(2H，d，J＝8Hz)，7.94(2H，d，J＝8Hz).

Embodiment 14

4-[3-(2,4-diamino-7H-pyrrolo-[2,3-d] pyrimidine-5-yl) propyl group] preparation of ethyl benzoate

Add ethanol (2.0ml) and the water (0.02ml) that contains 20% (w/w) hydrochloric acid in the suspension that in ethanol, forms toward the product (404mg) of example 13.And at room temperature stirred the mixture 2 hours, water (10ml) diluted reaction mixture and the past ammoniacal liquor that wherein adds make it be alkalescence, then remove most of ethanol by underpressure distillation, filter and collect the precipitation that obtains, water successively.Ethanol and ether washing, drying obtains title compound (300mg).

IR(KBr)：3330，3225，2930，1705，1610，1575，1490，1450，1410，1280，1180，1105，1020，830cm ^1-

¹H-NMR(CDCl ₃/CD ₃OD)：1.39(3H，t，J＝7.2Hz)，2.05(2H，m)，2.67(2H，t，J＝7.2Hz)，2.78(2H，t，J＝7.2Hz)，4.37(2H，q，J＝7.2Hz)，6.50(1H，s)，7.27(2H，d，J＝8Hz)，7.98(2H，d，J＝8Hz).

Example 15

4-[3-(2,4-diamino-7H-pyrrolo-[2,3-d] pyrimidine-5-yl) propyl group] benzoic preparation

With the product (340mg) of example 14 be suspended in tetrahydrofuran (THF)-water mixing solutions (5: 1,60ml) in, toward the aqueous solution (4.0ml) that wherein adds 1N sodium hydroxide, and stirred the mixture 18 hours at 50 ℃.Underpressure distillation to be removing most of tetrahydrofuran (THF) from mixture, toward wherein add 1N hydrochloric acid with in and resistates.Filter and collect the precipitation that obtains, water, methyl alcohol and ether washing successively, drying under reduced pressure obtains title compound (305mg).

IR(KBr)：3480，3390，3130，2940，1650，1605，1550，1460，1390，1255，1180，1095，980，780cm ^-1

¹H-NMR(Me ₂SO-d ₆)：1.73-1.95(2H，m)，2.67(2H，t，J＝7Hz)，2.71(2H，t，J＝7Hz)，5.37(2H，s)，5.95(2H，s)，6.43(1H，s)，7.33(2H，d，J＝8Hz)，7.86(2H，d，J＝8Hz)，10.40(1H，s).

Example 16

N-[4-[3-(2,4-diamino-7H-pyrrolo-[2,3-d] pyrimidine-5-yl) propyl group] benzoyl]-preparation of L-glutamate diethyl ester

The product (339mg) and the L-diethyl glutamate hydrochloride (748mg) of example 15 are suspended in the dimethyl formamide (4.0ml), also stir toward wherein adding the solution (858mg) of diphenylphosphine acyl azide (858mg) in dimethyl formamide at 0 ℃.Then under identical temperature toward wherein dripping the solution (4.0ml) of triethylamine in dimethyl formamide, stirred the mixture 30 minutes at 0 ℃.At room temperature stirred then 63 hours, then underpressure distillation is desolvated to remove.With column chromatography (15g silica gel is from the isolated methylene dichloride of strong aqua: from the isolated methylene dichloride of strong aqua-ethanol=40: 1 → 30: the 1) resistates that extraction obtains, obtain colourless crystalline title compound (386mg).

The IR of compound (KBr) and ¹H-NMR (Me ₂SO-d ₆) identical with the data of the product of example 6

Embodiment 17

N-[4-[4-(triaminopyrimidine-5-yl)-5,5-dimethoxy amyl group] benzyl]-preparation of L-glutamate diethyl ester

With the product (403mg) of example 13 be suspended in tetrahydrofuran (THF)-water mixing solutions (5: 1,8.0ml) in, spend the night toward wherein adding the solution (2.0ml) of 1N sodium hydroxide and stirring the mixture at 40 ℃.After adding 1N hydrochloric acid (2.0ml) neutralise mixt, except that desolvating, dried residue obtains thick 4-[4-(triaminopyrimidine-5-yl)-5,5-dimethoxy amyl group with the method for underpressure distillation] phenylformic acid.All thick products and diethyl glutamate hydrochloride (360mg) are suspended in the dimethyl formamide (4.0ml), 0 ℃ toward wherein adding the solution (4.0ml) of cyano group phosphonic acids di-alcohol (171mg) in dimethyl formamide, and stir the mixture.Then under identical temperature toward wherein dripping the solution (4.0ml) of triethylamine (303mg) in dimethyl formamide, and stirred the mixture 30 minutes at 0 ℃, at room temperature stirred then 3 hours, then by underpressure distillation except that desolvating.Purify to obtaining resistates with flash distillation column chromatography (15g silica gel, methylene chloride-methanol=30: 1 → 5: 1), obtain title compound (417mg).

¹H-NMR(CDCl ₃/CD ₃OD)：1.22(3H，t，J＝7Hz)，1.32(3H，t，J＝7Hz)，1.92-1.74(2H，m)，1.93-2.38(4H，m)，2.45-2.56(2H，m)，2.63(2H，t，J＝7.4Hz)，2.77-2.87(1H，m)，3.48(3H，s)，3.51(3H，s)，4.12(2H，q，J＝7Hz)，4.23(2H，q，J＝7Hz)，4.35(′H，d，J＝3.2Hz)，4.77-4.85(1H，m)，7.21(1H，d，J＝8.4Hz)，7.94(2H，d，J＝8.4Hz).

Embodiment 18

N-[4-[3-(2,4-diamino-7H-pyrrolo-[2,3-d] pyrimidine-5-yl) propyl group] benzoyl]-preparation of L-glutamate diethyl ester

Add ethanol (2.0ml) and the water (0.02ml) that contains 20% (w/w) hydrochloric acid in the suspension that forms in ethanol (2.0ml) toward the product (100mg) of example 17, at room temperature heated mixt is 2 hours.Water (10ml) diluted reaction mixture neutralizes toward wherein adding ammoniacal liquor, then desolvates to remove by underpressure distillation.With column chromatography (15g silica gel; From the isolated methylene dichloride of strong aqua: from the isolated methylene dichloride of strong aqua-ethanol=40: 1 → 30: 1) resistates of purifying and obtaining, obtain title compound (69mg).

The IR of compound (KBr) and ¹H-NMR (Me ₂SO-d ₆) identical with the data of the product of example 6

Embodiment 19

4-[3-(2,4-diamino-7H-pyrrolo-[2,3-d]-pyrimidine-5-yl) propyl group] benzoic preparation

With successively according to the step of reference example 3, reference example 2 and example 13,4-[4-(2 by 4-(4-butyl phenyl ether methyl-formiate) acquisition, 4,6-Triaminopyrimidine-5-yl)-5,5-dimethoxy amyl group] methyl benzoate (2.15g) is dissolved in the mixing solutions of 1N tetrahydrofuran (THF) (15.5ml) and 1N hydrochloric acid (7.1ml), and stirred the mixture 1 hour at 68 ℃.In this step, do not separate 4-[3-(2,4-diamino-7H-pyrrolo-[3, the 2-d] pyrimidine-5-yl) propyl group that makes in the reaction mixture] methyl benzoate.The aqueous solution (2.85ml) that directly adds methyl alcohol (5.7ml) and sodium hydroxide (0.684g) in the reaction mixture was 672 hydrolysis 1 hour.Remove organic solvent in the reaction mixture with distillation under vacuum, residual pH value of aqueous solution is adjusted to 3 with 6N hydrochloric acid.Filter to collect the precipitation that obtains, with less water washing, drying under reduced pressure, obtain title compound (1.72g) compound IR (KBr) and ¹H-NMR (Me ₂SO-d ₆) identical with the data of the product of example 19.

Press and example 1 to 19 described same procedure, can synthesize following compounds:

(1) N-[4-[3-(2-amino-4-hydroxy-7H-pyrrolo-[2,3-d]-pyrimidine-5-yl) propyl group] benzoyl]-L-L-glutamic acid benzhydryl ester,

(2) N-[4-[3-(2-amino-4-hydroxy-7H-pyrrolo-[2,3-d]-pyrimidine-5-yl) propyl group] benzoyl]-L-L-glutamic acid,

(3) N-[4-[3-(2-amino-4-hydroxy-6,7-H-dihydro-5H-pyrrolo-[2,3-d]-pyrimidine-5-yl) propyl group] benzoyl]-L-L-glutamic acid,

(4) N-[4-[3-(2-amino-4-sulfydryl-7H-pyrrolo-[2,3-d]-pyrimidine-5-yl) propyl group] benzoyl]-L-L-glutamic acid two (to methoxy-benzyl) ester,

(5) N-[4-[3-(2-amino-4-sulfydryl-7H-pyrrolo-[2,3-d]-pyrimidine-5-yl) propyl group] benzoyl]-L-L-glutamic acid,

(6) N-[4-[3-(2-amino-4-sulfydryl-6,7-dihydro-5H-pyrrolo-[2,3-d]-pyrimidine-5-yl) propyl group] benzoyl]-L-L-glutamic acid,

(7) N-[4-[3-(2,4-diamino-6-methyl-7H-pyrrolo-[2,3-d]-pyrimidine-5-yl) propyl group] benzoyl]-L-L-glutamic acid,

(8) N-[4-[3-(2,4-diamino-6-methyl-6,7-dihydro-5H-pyrrolo-[2,3-d] pyrimidine-6-yl) propyl group] benzoyl]-L-L-glutamic acid,

(9) N-[4-[3-(2,4-diamino-6-ethyl-7H-pyrrolo-[2,3-d] pyrimidine-5-yl) propyl group] benzoyl]-L-L-glutamic acid,

(10) N-[4-[3-(2,4-diamino-6-ethyl-6,7-dihydro-5H-pyrrolo-[2,3-d] pyrimidine-5-yl) propyl group] benzoyl]-L-L-glutamic acid,

(11) N-[4-[3-(2,4-diamino-6-vinyl-7H-pyrrolo-[2,3-d] pyrimidine-5-yl) propyl group] benzoyl]-L-L-glutamic acid,

(12) N-[4-[3-(2,4-diamino-6-ethynyl-7H-pyrrolo-[2,3-d] pyrimidine-5-yl) propyl group] benzoyl]-L-L-glutamic acid,

(13) N-[4-[3-(2,4-diamino-6-hydroxymethyl-7H-pyrrolo-[2,3-d] pyrimidine-5-yl) propyl group] benzoyl]-L-L-glutamic acid,

(14) N-[4-[3-(2,4-diamino-7H-pyrrolo-[2,3-d] pyrimidine-5-yl)-1-methyl-propyl] benzoyl]-L-L-glutamic acid,

(15) N-[4-[3-(2,4-diamino-6,7-dihydro-5H-pyrrolo-[2,3-d] pyrimidine-5-yl)-1-methyl-propyl] benzoyl]-L-L-glutamic acid,

(16) N-[4-[3-(2,4-diamino-7H-pyrrolo-[2,3-d] pyrimidine-5-yl)-1-hydroxymethyl propyl group] benzoyl]-L-L-glutamic acid,

(17) N-[4-[3-(2,4-diamino-7H-pyrrolo-[2,3-d] pyrimidine-5-yl)-1-(formyl methyl) propyl group] benzoyl]-L-L-glutamic acid,

(18) N-[4-[3-(2,4-diamino-7H-pyrrolo-[2,3-d] pyrimidine-5-base-1-chloromethyl propyl group] benzoyl]-L-L-glutamic acid,

(19) N-[4-[3-(2,4-diamino--7H-pyrrolo-[2,3-d] pyrimidine-5-yl)-1-dichloromethyl propyl group] benzoyl]-L-L-glutamic acid,

(20) N-[4-[3-(2,4-diamino-7H-pyrrolo-[2,3-d] pyrimidine-5-yl)-1-trifluoromethyl propyl group] benzoyl]-L-L-glutamic acid,

(21) N-[4-[3-(2,4-diamino-7H-pyrrolo-[2,3-d] pyrimidine-5-yl)-1-methoxymethyl propyl group] benzoyl]-L-L-glutamic acid,

(22) N-[4-[3-(2,4-diamino-7H-pyrrolo-[2,3-d] pyrimidine-5-yl)-1-ethoxyl methyl propyl group] benzoyl]-L-L-glutamic acid,

(23) N-[4-[3-(2,4-diamino-7H-pyrrolo-[2,3-d] pyrimidine-5-yl)-1-cyano methyl propyl group] benzoyl]-L-L-glutamic acid,

(24) N-[4-[3-(2,4-diamino-7H-pyrrolo-[2,3-d] pyrimidine-5-yl)-1-methylthiomethyl) propyl group] benzoyl]-L-L-glutamic acid,

(25) N-[4-[3-(2,4-diamino-7H-pyrrolo-[2,3-d] pyrimidine-5-yl)-2-first third propyl group] benzoyl]-L-L-glutamic acid, (26) N-[4-[3-(2,4-diamino-7H-pyrrolo-[2,3-d] pyrimidine-5-yl)-and the 3-methyl-propyl] benzoyl]-L-L-glutamic acid

(27) N-[4-[3-(2,4-diamino-7H-pyrrolo-[2,3-d] pyrimidine-5-yl)-1, the 1-dimethyl propyl] benzoyl]-L-L-glutamic acid,

(28) N-[4-[3-(2,4-diamino-6,7-dihydro-5H-pyrrolo-[2,3-d] pyrimidine-5-yl)-1, the 1-dimethyl propyl] benzoyl]-L-L-glutamic acid,

(29) N-[4-[3-(2,4-diamino-7H-pyrrolo-[2,3-d] pyrimidine-5-yl)-2, the 2-dimethyl propyl] benzoyl]-L-L-glutamic acid, and (30) N-[4-[3-(2,4-diamino-7H-pyrrolo-[2,3-d] pyrimidine-5-yl)-the 1-ethyl propyl] benzoyl]-L-L-glutamic acid

(31) N-[4-[3-(2,4-diamino-6,7 dihydros-5H-pyrrolo-[2,3-d] pyrimidine-5-base-1-ethyl propyl benzoyl]-L-L-glutamic acid,

(32) N-[4-[3-(2,4-diamino-7H-pyrrolo-[2,3-d] pyrimidine-5-yl)-1-hydroxyethyl propyl group] benzoyl]-L-L-glutamic acid,

(33) N-[4-[3-(2,4-diamino-7H-pyrrolo-[2,3-d] pyrimidine-5-yl)-1-(2-formyl radical ethyl) propyl group] benzoyl]-L-L-glutamic acid,

(34) N-[4-[3-(2,4-diamino-7H-pyrrolo-[2,3-d] pyrimidine-5-yl)-1-methoxy ethyl propyl group] benzoyl]-L-L-glutamic acid,

(35) N-[4-[3-(2,4-diamino-7H-pyrrolo-[2,3-d] pyrimidine-5-yl)-1-ethoxyethyl group propyl group] benzoyl]-L-L-glutamic acid,

(36) N-[4-[3-(2,4-diamino-7H-pyrrolo-[2,3-two pyrimidines-5-yl)-2-ethyl propyl] benzoyl]-L-L-glutamic acid,

(37) N-[4-[3-(2,4-diamino-7H-pyrrolo-[2,3-d] pyrimidine-5-yl)-1-vinyl propyl group] benzoyl]-L-L-glutamic acid,

(38) N-[4-[3-(2,4-diamino-7H-pyrrolo-[2,3-d] pyrimidine-5-base-2-vinyl propyl group] benzoyl]-L-L-glutamic acid,

(39) N-[4-[3-(2,4-diamino-7H-pyrrolo-[2,3-d] pyrimidine-5-base-1-allyl group propyl group] benzoyl]-L-L-glutamic acid,

(40) N-[4-[3-(2,4-diamino-7H-pyrrolo-[2,3-d] pyrimidine-5-yl)-1-ethynyl propyl group] benzoyl]-L-L-glutamic acid,

(41) N-[4-[3-(2,4-diamino-7H-pyrrolo-[2,3-d] pyrimidine-5-yl)-1-(sec.-propyl) propyl group] benzoyl]-L-L-glutamic acid,

(42) N-[4-[3-(2,4-diamino-7H-pyrrolo-[2,3-d] pyrimidine-5-yl)-1-(2-propenyl) propyl group] benzoyl]-L-L-glutamic acid,

(43) N-[4-[3-(2,4-diamino-7H-pyrrolo-[2,3-d] pyrimidine-5-yl)-1-(alkynes third-1-yl) propyl group] benzoyl]-L-L-glutamic acid,

(44) propyl group N-[4-[3-(2,4-diamino-7H-pyrrolo-[2,3-d] pyrimidine-5-yl)-1-2-proyl)] benzoyl]-L-L-glutamic acid,

(45) 4-[4-(triaminopyrimidine-5-yl)-5,5-dimethoxy amyl group] methyl benzoate,

(46) 4-[4-(triaminopyrimidine-5-yl)-5,5-dimethoxy amyl group] the phenylamino benzoic acid methyl esters,

(47) 4-[4-(triaminopyrimidine-5-yl)-5,5-two (first sulfydryl) amyl group] phenylformic acid is to the anisole methyl esters,

(48) N-[4-[4-(triaminopyrimidine-5-yl)-5,5-dimethoxy amyl group] benzoyl]-L-L-glutamic acid benzhydryl ester,

(49) 4-[3-(2,4-diamino-7H-pyrrolo-[2,3-d] pyrimidine-5-base propyl group] methyl benzoate,

(50) 4-[3-(2,4-diamino-7H-pyrrolo-[2,3-d] pyrimidine-5-yl) propyl group] the phenylamino benzoic acid methyl esters,

Claims (7)

1. method for preparing the compound or its salt shown in the following general formula,

Wherein encircle the pyrrole ring that the A representative can be hydrogenated; X represents amino or hydroxyl; R ¹, R ²And R ³Can be identical or different, represent hydrogen or low alkyl group respectively; R ⁴Represent OR ⁵, R wherein ⁵Represent hydrogen or low alkyl group, benzyl, NHCH (COOR ⁶) CH ₂CH ₂COOR ⁷, R wherein ⁶And R ⁷Represent hydrogen separately, low alkyl group or benzyl; N represents 1 to 4 integer,

It is characterized in that this method comprises makes the compound or its salt shown in the following general formula

X wherein, R ¹, R ², R ³, R ⁴Identical with n with above-mentioned definition; Y ¹And Y ²The Sauerstoffatom of respectively doing for oneself; R ⁸And R ⁹Be low alkyl group,

Contact with the acid in the water-containing organic solvent.

2. the described method of claim 1, wherein aqueous organic solvent is the mixture of a kind of alcohol, acetic ester, ether, ketone, acid amides, sulfoxide or nitrile and water.

3. the described method of claim 1, wherein acid is a kind of mineral acid.

4. the described method of claim 1, wherein R ¹, R ²And R ³It respectively is hydrogen atom.

5. the described method of claim 1, wherein Y ¹And Y ²Be Sauerstoffatom.

6. the described method of claim 1, wherein R ⁸And R ⁹Respectively be C _1-3Alkyl.

7. the described method of claim 1, wherein R ⁵, R ⁶And R ⁷Respectively be C _1-5Alkyl or benzyl.