CN102659785B - 2-amino-6-aminomethylpurine compound and preparation method and application thereof - Google Patents
2-amino-6-aminomethylpurine compound and preparation method and application thereof Download PDFInfo
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- 0 CCCc([n](*(C)C1CC1)c1n2)nc1c(NCC)nc2F Chemical compound CCCc([n](*(C)C1CC1)c1n2)nc1c(NCC)nc2F 0.000 description 2
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Abstract
The invention discloses a 2-amino-6-aminomethylpurine compound and a preparation method and an application thereof. The compound has a structure shown in a general formula I. As proved by an activity screening experiment, the 2-amino-6-aminomethylpurine compound has suppressing activity on cyclin-dependent kinases and proliferation of a plurality of tumor cells, and can be possibly used for treating diseases caused by activity imbalance of cyclin-dependent kinases. The invention further relates to a pharmaceutical application of a composition containing the compound shown as the general formula I. The general formula I is shown in the description.
Description
Technical field
The present invention relates to 2-amino-6-aminomethyl purine compound and its preparation method and application, belong to technical field of chemistry.
Background technology
Malignant tumour, is called again cancer, is the disease of serious harm human life health.According to the World Health Organization (WTO), in the < < world cancer of issue in 2008, report > >, in worldwide in 2008, there are 1,200 ten thousand people to be made a definite diagnosis and suffer from cancer, and also have 7,600,000 people because of cancer death, this death toll has accounted for 13% of whole world death toll, the number of estimating global range internal cause cancer mortality also can continue to increase, if cancer is not intervened, the year two thousand thirty will have 1,200 ten thousand people to die from cancer, this wherein 70% death be to occur in low income or middle-income country.Therefore, malignant tumour has become the common significant problem of being concerned about of the universe.Grasp the pathogenic factor of malignant tumour and the molecular mechanism developing occurs, the selectivity cancer therapy drug of finding high-efficiency low-toxicity has become the significant task of medical field research.Malignant tumour is under environmental factors and inherited genetic factors acting in conjunction, causes the class disease due to cell cycle disorder, cell property growth out of control.Since the molecular mechanism of cell cycle regulation is since early 1970s is found, cell cycle chechpoint just becomes the important target spot of antitumor drug research.Cell cycle protein dependent kinase (CDKs) is the main regulon of cell cycle, suppresses CDKs and can effectively block tumour cell cycle, suppresses propagation and the differentiation of cell, for the treatment of tumour provides new thinking.
Cell cycle protein dependent kinase (CDKs) is the protein kinase of a class cyclin dependent (Cyclin), belong to Serine and threonine kinase enzyme family (referring to Nurse, P., Nature, 1975,256,547-551.).The relative molecular weight of CDKs is at 35kD to (comprising 200 to 400 amino acid) between 45KD, and the homology of the CDK of different subtype is about 40-75%, all contains approximately 300 amino acid whose conservative property catalytic centers, and this is the same with other protein kinase.Up to now, in human genome, have been found that 13 CDK hypotypes (CDK1-13) and the cyclins more than 25 kinds.CDKs need to be combined its activity of rear competence exertion with regulating subunit cyclin, and the effect of different CDK/cyclin mixtures phase when cell cycle different be different (referring to Wang, Q., et al., Curr Med.Chem., 2011,18,2025-2043).According to classical cell cycle model, participate in directly the cell cycle when each the CDKs/cyclin mixture of phase transformation be CDK4 (6)/cyclin D, CDK2/cyclin E, CDK2/cyclin A and CDK1/cyclin B.Cyclin D plays an important role in the G1 phase after being combined with CDK4 and CDK6; CDK2/cyclin E can impel cell to enter the S phase from the G1 phase; Cyclin A can impel the cell cycle to complete the conversion of S phase after being combined with CDK2 and CDK1 respectively, prepares to enter the M phase; The M phase enter and regulate be under the control of CDK1/cyclinB, complete (referring to van den Heuvel, S.; Et al., Science, 1993,262,2050-2054 and Hochegger, H., et al., Nat Rev Mol Cell Biol, 2008,9,910-916.).
The imbalance of cell cycle is the important symbol of human cancer, and in its setting device, the change of moiety is relevant with the generation of most of mankind's tumour.Therefore, CDKs is considered to the potential target spot of new generation anti-cancer medicament.Developing some the specific C DKs inhibitor that can block cancer cell cycle and cell death inducing should be very effective for the treatment of cancer.In the past few decades, developed the CDKs inhibitor of numerous structure types, and the kind scope of inhibitor is also in continuous expansion, has comprised some natural products and derivative thereof.Add developing rapidly of medicinal design, the X-ray crystal structure forming by a large amount of CDKs and inhibitor is carried out rational medicinal design, the competitive binding mode of inhibitor is not only provided, can also have observed the active binding site of micromolecular inhibitor.Up to now, existing more than ten research that compound enters clinical I phase or II phase.
The present invention is according to the Three Dimensions Structure of CDK2, based on the method for Computer-Aided Drug Design, taking into full account on the basis of the binding pattern of ATP-binding site in 2-amino-6-aminomethyl purine compound and CDK2, designed a series of small molecules CDK inhibitor, then carry out chemosynthesis and screening active ingredients, obtained thering is the lead compound that exploitation is worth.
Summary of the invention
The present invention is directed to the deficiencies in the prior art, a kind of 2-amino-6-aminomethyl purine compound and preparation method thereof is provided; The present invention also provides the application of this compounds.
Technical scheme of the present invention is as follows:
One, 2-amino-6-aminomethyl purine compound of the present invention
2-amino-6-aminomethyl purine compound or its pharmacy acceptable salt, steric isomer, solvate or prodrug, have the structure shown in formula I:
Wherein,
R1 is aryl, heteroaryl, and assorted alkyl, cycloalkyl, is optionally replaced by one or more following groups: hydroxyl, halogen, nitro, itrile group, carboxyl, alkyl, alkoxyl group, sulfoamido or methylsulfonic acid base;
R2 is aryl, benzyl, and alkyl, heteroaryl, is optionally replaced by one or more following groups: hydroxyl, halogen, nitro, itrile group, carboxyl, alkyl, alkoxyl group, sulfoamido or methylsulfonic acid base;
R3 is hydrogen, carboxyl or hydroxymethyl;
R4 is hydrogen, methyl, ethyl, sec.-propyl, cyclopropyl or cyclopentyl;
Described aryl refers to aromatic carbocyclic group, and aromatic ring contains 6-10 carbon atom;
Described heteroaryl is aromatic heterocycle, is monocycle or bicyclic radicals; Comprise thienyl, furyl, pyrryl, pyridyl, pyrazinyl, thiazolyl, pyrimidyl, quinolyl, tetrazole base, benzothiazolyl, benzofuryl or indyl;
That described assorted alkyl refers to is saturated or unsaturated, carbon atoms and at least one heteroatomic chain, and wherein any one heteroatoms is non-conterminous; In assorted alkyl, contain 2-15 carbon atom, preferably contain 2-10 atom; Assorted alkyl is straight or branched, replacement or unsubstituted;
Described cycloalkyl is replacement or unsubstituted, saturated or undersaturated cyclic group, and it contains carbon atom and/or one or more heteroatoms; This ring is monocycle or condensed ring, the ring system of bridged ring or volution; Monocycle has 3-9 atom, preferably has 4-7 atom, and many rings contain 7-17 atom, preferably contain 7-13 atom;
Described alkyl refers to the chain of saturated carbon atom, preferably contains 1-8 atom; Alkyl is straight or branched;
Described halogen comprises fluorine, chlorine, bromine or iodine.
Preferably, R
1for phenyl or cyclohexyl; R
2for substituted-phenyl or hydroxyl substituted alkyl; R
3for hydrogen, carboxyl or hydroxymethyl; R
4for hydrogen or sec.-propyl.
Further preferred, above-mentioned formula I compound is one of following:
N
6-benzyl-N
2-(4-bromo phenyl)-9H-purine-2,6-diamines (4a),
N
6-benzyl-N
2-(4-p-methoxy-phenyl)-9H-purine-2,6-diamines (4b),
N
6-benzyl-N
2-p-methylphenyl-9H-purine-2,6-diamines (4c),
N
6-benzyl-N
2-(3-methoxyphenyl)-9H-purine-2,6-diamines (4d),
4-((6-benzylamine-9H-purine-2)-amido) phenol (4e),
N
6-benzyl-N
2-(4-fluorophenyl)-9H-purine-2,6-diamines (4f),
4-((6-benzylamine-9H-purine-2)-amido) benzsulfamide (4g),
N
6-benzyl-N
2-phenyl-9H-purine-2,6-diamines (4h),
N
6-benzyl-N
2-(4-first sulfo group phenyl)-9H-purine-2,6-diamines (4i),
4-((6-cyclohexyl methylamine-9H-purine-2)-amido) benzsulfamide (4j),
N
6-cyclohexyl methyl-N
2-phenyl-9H-purine-2,6-diamines (4k),
4-(6-cyclohexyl methylamine)-9H-purine-2-amido) phenol (4l),
N
6-cyclohexyl methyl-N
2-(4-fluorophenyl)-9H-purine-2,6-diamines (4m),
N
6-cyclohexyl methyl-N
2-(4-tolyl)-9H-purine-2,6-diamines (4n),
N
6-cyclohexyl methyl-N
2-(4-p-methoxy-phenyl)-9H-purine-2,6-diamines (4o),
N
6-cyclohexyl methyl-N
2-(4-bromophenyl)-9H-purine-2,6-diamines (4p),
N
6-cyclohexyl methyl-N
2-(4-first sulfo group phenyl)-9H-purine-2,6-diamines (4q),
2-(6-(cyclohexyl methylamine base)-9-sec.-propyl-9H-purine-2-amido)-n-butyl alcohol (10a),
2-(6-(cyclohexyl methylamine base)-9-sec.-propyl-9H-purine-2-amido) ethanol (10b),
2-(6-(cyclohexyl methylamine base)-9-sec.-propyl-9H-purine-2-amido)-3-hydroxy-propionic acid (10c),
2-(6-(cyclohexyl methylamine base)-9-sec.-propyl-9H-purine-2-amido)-3-hydroxybutyrate (10d),
2-(6-(cyclohexyl methylamine base)-9-sec.-propyl-9H-purine-2-amino) propane-1,3-glycol (10e),
2-(6-(cyclohexyl methylamine base)-9-sec.-propyl-9H-purine-2-amido)-4-methylpentane-1-alcohol (10f),
2-(6-(cyclohexyl methylamine base)-9-sec.-propyl-9H-purine-2-amido)-3-methyl-1-butanol (10g),
2-(6-(cyclohexyl methylamine base)-9-sec.-propyl-9H-purine-2-amido) propane-1-alcohol (10h) or
2-(6-(cyclohexyl methylamine base)-9-sec.-propyl-9H-purine-2-amido) propionic acid (10i).
Term and definition implication used herein is as follows:
" pharmacy acceptable salt " refers to that compound of Formula I has curative effect and nontoxic salt form.It can form anion salt by arbitrary acidic-group (as carboxyl), or forms cationic salts by arbitrary basic group (as amino).Much such salt known in the art.At the upper cationic salts forming of any acidic-group (as carboxyl), or at the upper anion salt forming of any basic group (as amino).It is known in the art that these salt have many, as cationic salts comprises salt and the organic salt (as ammonium salt) of basic metal (as sodium and potassium) and alkaline-earth metal (as magnesium and calcium).Also can obtain easily anion salt by the I that uses corresponding acid treatment alkaline form, such acid comprises that mineral acid is as sulfuric acid, nitric acid, phosphoric acid etc.; Or organic acid is as acetic acid, propionic acid, oxyacetic acid, 2 hydroxy propanoic acid, Acetylformic acid, oxalic acid, propanedioic acid, succsinic acid, toxilic acid, fumaric acid, oxysuccinic acid, tartrate, 2-hydroxyl-1,2,3-, the third three acid, methylsulfonic acid, ethyl sulfonic acid, benzene methanesulfonic acid, 4-toluene sulfonic acide, cyclohexyl-sulfinic acid, 2 hydroxybenzoic acid, 4-amino-2-hydroxybenzoic acid etc.These salt are that those of skill in the art know, and those skilled in the art can prepare any salt that this area knowledge provides.In addition, those of skill in the art can get certain salt according to factors such as solubleness, stability, easy preparations and give up another kind of salt.The mensuration of these salt and optimization are within the scope of those of skill in the art's experience.
The present invention's " steric isomer " used defined the form of the compounds of this invention or all possible steric isomer of its physiological derivative.Unless otherwise directed, the chemical name of the compounds of this invention comprises the mixture of all possible stereochemical form, all diastereomers and enantiomorph that affiliated mixture comprises basic structure molecule, and the single isomeric forms of the compounds of this invention of substantially pure, wherein contain lower than 10%, preferably lower than 5%, particularly lower than 2%, most preferably lower than other isomer of 1%.The various stereoisomer forms of class peptide compounds of the present invention are all obviously contained in scope of the present invention.
" solvate " is the title complex that solute (as cell cycle protein dependent kinase inhibitor) and solvent (as water) are combined to form.Referring to J.Honig etc., The Van Nostrand Chemist ' s Dictionary, p.650 (1953).The pharmaceutically acceptable solvent that the present invention adopts comprises not bioactive those solvents of interference cell cyclin-dependent kinase inhibitor (for example, known to water, ethanol, acetic acid, DMF, dimethyl sulfoxide (DMSO) and this those skilled in the art or easy definite solvent).
The form of all right other the protected form of compound of Formula I or derivative exists, and these forms will be apparent to those skilled in the art, and all should be contained in scope of the present invention.
Substituting group as above self also can be replaced by one or more substituting groups.Such substituting group is included in C.Hansch and A.Leo, those substituting groups of listing in Substituent Constants for Correlation Analysis in Chemistry and Biology (1979).Preferred substituting group comprises alkyl, thiazolinyl, alkoxyl group, hydroxyl, oxygen base, nitro, amino, aminoalkyl group (as aminomethyl etc.), cyano group, halogen, carboxyl, carbonylic alkoxy (as carbonyl oxyethyl group etc.), sulfenyl, aryl, cycloalkyl, heteroaryl, Heterocyclylalkyl (as piperidyl, morpholinyl, pyrryl etc.), imino-, hydroxyalkyl, aryloxy, arylalkyl and combination thereof.
Two, the preparation method of 2-amino-6-aminomethyl purine compound of the present invention
The preparation method of 2-amino-6-aminomethyl purine compound of the present invention is one of following method:
Synthetic route one: take 2-amido-6-chloropurine as raw material, obtain 2-amino-6-substituted purin with the amine generation nucleophilic substitution reaction of various replacements, then at HBF
4and NaNO
2under effect, through Ba Erci-Xi Man (Balz-Schiemann) reaction, make the fluoro-6-substituted purin of 2-, finally react with the amine of various replacements again and generate target compound 4a-4i; Route one is as follows:
Wherein, described in the same general structure I of R1 and R2;
The amine of described various replacements is the amine being replaced by phenyl, heteroaryl, assorted alkyl or cycloalkyl, and in organic amine structure, can have hydroxyl, halogen, nitro, itrile group, carboxyl, alkyl, alkoxyl group, sulfoamido or methylsulfonic acid base.
Reagent and condition in said synthesis route one reaction formula: a. propyl carbinol, triethylamine, 130 ℃ of backflows; B.HBF
4, NaNO
2, H
2o ,-15 ℃; C. trifluoroacetic acid (TFA), propyl carbinol, 130 ℃ or trifluoroacetic acid (TFA), trifluoroethanol (TFE), 85 ℃.
The structural formula of the target compound of synthetic route one is as shown in table 1 below:
Table 1 compound 4a-4i structural formula
Synthetic route two: for raw material, and obtain the chloro-6-substituted purin of 2-after the amine generation nucleophilic substitution reaction of various replacements with 2,6-dichloropurine, and then react with the amine of various replacements and generate target compound 4j-4q; Route two is as follows:
Wherein, described in the same general structure I of R1 and R2;
The amine of described various replacements is the amine being replaced by phenyl, heteroaryl, assorted alkyl or cycloalkyl, and in organic amine structure, can have hydroxyl, halogen, nitro, itrile group, carboxyl, alkyl, alkoxyl group, sulfoamido or methylsulfonic acid base.
Reagent and condition in said synthesis route two reaction formula: d. propyl carbinol, triethylamine, 130 ℃ of backflows; E. trifluoroacetic acid, propyl carbinol, 130 ℃ or trifluoroacetic acid, trifluoroethanol, 85 ℃.
The structural formula of the target compound of synthetic route two is as shown in table 2 below:
Table 2 compound 4j-4q structural formula
Synthetic route three: take the fluoro-6-chloropurine of 2-as raw material, with the amine generation nucleophilic substitution reaction of various replacements, obtain fluoro-6 substituted purins of 2-, then by 9-H alkylation, finally react generation with each seed amino acid or amino alcohol and obtain target compound 10a-10i; Route three is as follows:
Wherein, the same general structure I of R1, R2, R3 and R4;
The amine of described various replacements is the amine being replaced by phenyl, heteroaryl, assorted alkyl or cycloalkyl, and in organic amine structure, can have hydroxyl, halogen, nitro, itrile group, carboxyl, alkyl, alkoxyl group, sulfoamido or methylsulfonic acid base.
Described each seed amino acid or amino alcohol are amino acid and the corresponding amino alcohol thereof that Serine, Threonine, L-Ala, α-amino-isovaleric acid, leucine etc. are conventional.
Reagent and condition in said synthesis route three reaction formula: f. propyl carbinol, triethylamine, 130 ℃ of backflows; G. haloalkane, K
2cO
3, dimethyl sulfoxide (DMSO); H. amino acid, N-Methyl pyrrolidone, 1,8-diazabicylo [5.4.0], 11 carbon-7-alkene (DBU), 160 ℃ or amino alcohol, dimethyl sulfoxide (DMSO), diisopropylethylamine, 120-160 ℃.
The structural formula of the target compound of synthetic route three is as shown in table 3 below:
Table 3 compound 10a-10i structural formula
The concrete operation step of described compound will be described in detail in an embodiment.
Three, the pharmaceutical composition that contains the compounds of this invention
One is suitable for the oral mammiferous pharmaceutical composition that gives, the arbitrary compound that comprises above-mentioned general formula I and one or more pharmaceutically acceptable vehicle.
One is suitable for parenteral and gives mammiferous pharmaceutical composition, the arbitrary compound that comprises above-mentioned general formula I and one or more pharmaceutically acceptable vehicle.
Part derivative of the present invention can free form or is existed with salt form.Pharmacy acceptable salt of the known chemical compound lot type of those skilled in the art and preparation method thereof.Pharmacy acceptable salt comprises conventional avirulent salt, comprises such compound alkali and quaternary ammonium salt inorganic or that organic acid forms.
Compound of the present invention can form hydrate or solvate.The known hydrate that compound is formed during freeze-drying together with water of one skilled in the art or form the method for solvate when concentrated with suitable organic solvent in solution.
The present invention comprises the medicine that contains therapeutic dose the compounds of this invention, and the pharmaceutical composition of one or more pharmaceutically acceptable carriers and/or vehicle.Carrier comprises as salt solution, buffer saline, and glucose, water, glycerine, ethanol and their binding substances, below discuss in more detail.If needed, said composition can also comprise wetting agent or emulsifying agent in a small amount, or pH buffer reagent.Said composition can be liquid, suspension, emulsion, tablet, pill, capsule, extended release preparation or powder.Said composition can be mixed with suppository as triglyceride with traditional tamanori and carrier.Oral preparations can comprise that standard vector is as the mannitol of medicine grade, lactose, starch, Magnesium Stearate, soluble saccharin, Mierocrystalline cellulose and magnesiumcarbonate etc.Optionally preparation and determining, preparation can design mixing, granulation and compression or solvent components.In another approach, said composition can be mixed with nano particle.
The pharmaceutical carrier using can be solid or liquid.
Typical solid carrier comprises lactose, terra alba, sucrose, talcum, gel, agar, pectin, gum arabic, Magnesium Stearate, stearic acid etc.Solid carrier can comprise that one or more may be simultaneously as sweetener, lubricant, solubilizing agent, suspension agent, filler, glidant, compression aid, the material of tackiness agent or tablet-disintegrating agent; It can also be encapsulating material.In powder, carrier is pulverizing solid, and it mixes with pulverizing activeconstituents.In tablet, activeconstituents mixes with suitable ratio with the carrier with necessary compression property, with shape and the size compression of needs.Powder and tablet preferably comprise 99% activeconstituents at the most.Suitable solid carrier comprises, for example, and calcium phosphate, Magnesium Stearate, talcum, sugar, lactose, dextrin, starch, gel, Mierocrystalline cellulose, methylcellulose gum, sodium carboxymethyl-cellulose, polyvinylpyrrolidone alkane ketone, low melt wax and ion exchange resin.
Typical liquid vehicle comprises syrup, peanut oil, and sweet oil, water, etc.Liquid vehicle is for the preparation of solution, suspension, emulsion, syrup, the composition of tincture and sealing.Activeconstituents can dissolve or be suspended in pharmaceutically acceptable liquid vehicle as water, organic solvent, the mixture of the two or pharmaceutically acceptable oils or fat.Liquid vehicle can comprise other suitable medicated premixs as solubilizing agent, emulsifying agent, and buffer reagent, sanitas, sweetener, sweetener, suspension agent, thickening material, pigment, viscosity modifier, stablizes shape or osmotic pressure-conditioning agent.The suitable example that is used for the liquid vehicle of oral and administered parenterally comprises that water (partly comprises as above-mentioned additive, for example derivatived cellulose, preferably carboxymethyl cellulose sodium salt solution), alcohol (comprises monohydroxy-alcohol and polyvalent alcohol, for example ethylene glycol) and their derivative, and oils (for example fractionated coconut oil and peanut oil).Carrier for administered parenterally can also be that grease is as ethyl oleate and sec.-propyl myristate.Aseptic liquid vehicle is for the aseptic fluid composition of administered parenterally.Liquid vehicle for pressurized compositions can be halohydrocarbon or other pharmaceutically acceptable propelling agents.Sterile solution or aaerosol solution composition of liquid medicine can be used for, for example, and intravenously, intramuscular, intraperitoneal or subcutaneous injection.During injection, can push or inject gradually by single, entering the interior perfusion of passages through which vital energy circulates of 30 minutes.This compound can also be with the form oral administration of liquid or solids composition.
Carrier or vehicle can comprise time lag material known in the art, as glyceryl monostearate or distearin, also can comprise wax, ethyl cellulose, Vltra tears, methyl methacrylate etc.When preparation is when oral, generally acknowledge PHOSALPG-50 (phospholipid and 1,2-propylene glycol is concentrated, A.Nattermann & Cie.GmbH) in 0.01% tween 80 for the preparation of the acceptable oral preparations of other compounds, can be adapted to the preparation of the various compounds of the present invention.
While giving the compounds of this invention, can use medicament forms miscellaneous.If use solid carrier, preparation can be tablet, is placed into powder or piller form or lozenge or lozenge form in hard capsule.The amount of solid carrier changes to a great extent, but preferably from about 25mg to about 1.0g.If use liquid vehicle, preparation can be syrup, emulsion, soft capsule, aseptic injectable solution or suspension in the liquid suspension of ampoule or bottle or non-water.
In order to obtain stable water miscible formulation, compound or its pharmacy acceptable salt can be dissolved in to organic or inorganic aqueous acid, 0.3M succsinic acid or citric acid solution.Optionally, acid derivative can be dissolved in suitable basic solution.If can not get soluble form, compound can be dissolved in to suitable cosolvent or their combination.The example of suitable cosolvent like this includes but are not limited to, and concentration range is from the ethanol of 0-60% cumulative volume, propylene glycol, Liquid Macrogol, polysorbate 80, glycerine, polyoxyethylene fatty acid ester, fatty alcohol or glycerine hydroxy fatty acid ester etc.
Various release systems are known and can be for the administrations of compound or other various preparations, and these preparations comprise tablet, capsule, and injectable solution, the capsule in liposome, particulate, microcapsule, etc.The method of introducing includes, but are not limited to skin, intracutaneous, intramuscular, endoperitoneal, intravenous, subcutaneous, nasal cavity, lung, peridural, eyes and (conventionally preferred) oral route.Compound can be by administration easily any or that other is suitable, for example, by injecting or bolus injection, by epithelium or mucous membrane circuit (for example, oral mucosa, rectum and intestinal mucosa, etc.) absorb or the support by carrying medicament and can be in other biological promoting agent together administration.Can whole body or topical.For nose, when the treatment of segmental bronchus or lung disease or prevention, preferred route of administration is oral, nasal administration or segmental bronchus smoke substance or atomizer.
Four, application
The application of compound of the present invention in the medicine of preparing the mammalian diseases that prevention or treatment are relevant to the active imbalance of cell cycle protein dependent kinase.The described mammalian diseases relevant to the active imbalance of cell cycle protein dependent kinase comprises cancer, neurodegenerative disease, malaria and diabetes etc.
Embodiment
Below in conjunction with embodiment, the present invention will be further described, but be not limited to this.
The preparation of embodiment 1, compound 4a-4i
(1) N
6-benzyl-9H-purine-2,6-diamines (2)
By 2-amido-6-chloropurine (10.0g, 59.0mmol) and propyl carbinol (150mL) add in eggplant-shape bottle, then under agitation condition, add successively benzylamine (9.7mL, 88.5mmol) and triethylamine (20mL, 147.5mmol), finish, mixed solution is placed in 130 ℃ of oil baths, prolong cooling for reflux.After TLC detects raw material total overall reaction, stop refluxing, reaction solution is cooling, and decompression and solvent recovery obtains off-white color solid, after solid fully washs with n-propyl alcohol again, suction filtration, collects solid, obtains white powder 13.94g (58mmol after vacuum-drying, 98.4%), mp 234-238 ℃ of .HRMS:m/z calcd for C
12h
12n
6[M+1]
+241.1201Found:241.1218;
1h-NMR (600MHz, DMSO-d
6): δ ppm 4.64 (s, 2H); (5.75 s, 2H); (7.20 t, 2H); (7.28 t, 2H); (7.34 d, 2H); (7.68 s, 1H); (12.15 s, 1H)
(2) N-benzyl-2-fluoro-9H-purine-6-amine (3)
Compound 3 (2.4g, 10mmol) is used HBF
4(30mL) after dissolving, reaction solution is cooled to-15 ℃.At this temperature, use constant pressure funnel slowly to drip NaNO in reaction solution
2the aqueous solution of (1.4g, 20mmol), after dropwising,-15 ℃ are continued to stir 30min, then use 50%NaOH (w/v) to adjust pH to 7, separate out a large amount of solids, filter, filtrate discards, and solid adds in ethyl acetate, stirs after 10min, again filter, solid is used after ethyl acetate repetitive scrubbing, combined ethyl acetate, anhydrous magnesium sulfate drying.The crude product that decompression and solvent recovery obtains obtains white solid powder 0.83g (3.4mmol, 34%), mp 225-227 ℃ after purification by silica gel column chromatography.HRMS:m/z?calcd?for?C
12H
10FN
5[M+1]
+244.0998Found:244.0997;
1H-NMR(600MHz,DMSO-d
6):δppm?4.64(d,2H);7.24(d,1H);7.31-7.36(m,4H);8.11(s,1H);8.80(s,1H);13.05(1H,s).
(3) N
6-benzyl-N
2-(4-bromo phenyl)-9H-purine-2,6-diamines (4a)
Compound 3 (0.57g, 2.34mmol), para-bromoaniline (2.82g, 16.4mmol), trifluoroacetic acid (0.1mL, 1.3mmol), adds in propyl carbinol (10mL)., mixed solution is placed in 130 ℃ of oil baths, prolong cooling for reflux.TLC monitoring, treats compound 3 after completion of the reaction, stops refluxing, and reaction solution is cooling, separates out a large amount of solids.Suction filtration, methylene dichloride or methanol wash for solid, dry, crude product obtains brown color solid 0.5g (1.6mmol, 83%), mp 261-262 ℃ of .HRMS:m/z calcd for C after purification by silica gel column chromatography
18h
16n
6[M+1]
+317.1514Found:317.1521;
1h-NMR (600MHz, DMSO-d
6): δ ppm4.74 (s, 2H); (6.88 t, 1H); (7.20 t, 2H); (7.25 d, 1H); (7.33 t, 2H); (7.39 d, 2H); (7.70 d, 2H); (8.11 s, 1H); (8.32 s, 1H); (9.10 s, 1H); (12.81 s, 1H).
(4) N
6-benzyl-N
2-(4-p-methoxy-phenyl)-9H-purine-2,6-diamines (4b)
The same 4a of method.Compound 3 (0.25g, 1mmol), P-nethoxyaniline (0.19g, 1.5mmol), trifluoroacetic acid (0.4mL, 5mmol), propyl carbinol (10mL).Purification by silica gel column chromatography, obtains off-white color solid 0.11g (0.32mmol, 32%), mp 229-232 ℃ of .HRMS:m/z calcd for C
19h
18n
6o [M+1]
+347.1620Found:347.1610;
1h-NMR (600MHz, DMSO-d
6): δ ppm 3.69 (s, 3H); (4.69 s, 2H); (6.77 d, 2H); (7.20 t, 1H); (7.30 t, 2H); (7.37 d, 2H); (7.61 d, 2H); (7.78 s, 1H); (7.96 s, 1H); (8.60 s, 1H); (12.37 s, 1H).
(5) N
6-benzyl-N
2-p-methylphenyl-9H-purine-2,6-diamines (4c)
The same 4a of method.Compound 3 (0.61g, 2.5mmol), to monomethylaniline (1.88g, 17.6mmol), trifluoroacetic acid (0.1mL, 1.3mmol), propyl carbinol (20mL).Off-white color solid 0.39g (1.2mmol, 48%), mp 277-279 ℃ of .HRMS:m/z calcd for C
19h
18n
6[M+1]
+331.1671Found:331.1668;
1h-NMR (600MHz, DMSO-d
6): δ ppm 2.21 (s, 3H); (4.70 s, 2H); (6.97 d, 2H); (7.20 t, 1H); (7.30 t, 2H); (7.38 d, 2H); (7.61 d, 2H); (7.79 s, 1H); (8.01 s, 1H); (8.69 s, 1H); (12.40 s, 1H).
(6) N
6-benzyl-N
2-(3-methoxyphenyl)-9H-purine-2,6-diamines (4d)
The same 4a of method.Compound 3 (0.3g, 1.2mmol), m-anisidine (11g, 8.6mmol), trifluoroacetic acid (0.1mL, 1.3mmol), propyl carbinol (10mL).Purification by silica gel column chromatography, obtains white solid 0.18g (0.5mmol, 42%), mp 236-238 ℃ of .HRMS:m/z calcd for C
19h
18n
6o[M+1]
+347.1620Found:347.1614;
1h-NMR (600MHz, DMSO-d
6): δ ppm 3.68 (d, 3H); (4.72 s, 2H); (7.06 t, 1H); (7.20 t, 1H); (7.29 t, 3H); (7.39 d, 2H); (7.57 t, 2H); (7.82 s, 1H); (8.04 s, 1H); (8.81 s, 1H); (12.45 s, 1H).
(7) 4-((6-benzylamine-9H-purine-2)-amido) phenol (4e)
The same 4a of method.Compound 3 (0.4g, 1.6mmol), p-aminophenol (1.26g, 11.5mmol), trifluoroacetic acid (0.1mL, 1.3mmol), propyl carbinol (12mL).Purification by silica gel column chromatography obtains off-white color solid 0.22g (0.66mmol, 41%), mp258-260 ℃ of .HRMS:m/z calcd for C
18h
16n
6o[M+1]
+333.1464Found:333.1462;
1h-NMR (600MHz, DMSO-d
6): δ ppm 4.71 (t, 2H); (6.60 d, 2H); (7.20 t, 1H); (7.29 t, 2H); (7.37 d, 2H); (7.46 t, 2H); (7.75 s, 1H); (7.94 s, 1H); (8.45 s, 1H); (8.83 s, 1H); (12.32 s, 1H).
(8) N
6-benzyl-N
2-(4-fluorophenyl)-9H-purine-2,6-diamines (4f)
The same 4a of method.Compound 3 (0.46g, 1.9mmol), para-fluoroaniline (1.3mL, 13.2mmol), trifluoroacetic acid (0.1mL, 1.3mmol), propyl carbinol (10mL).White solid 0.13g (0.39mmol, 21%), mp 247-250 ℃ of .HRMS:m/z calcd for C
18h
15fN
6[M+1]
+335.1420Found:335.1350;
1h-NMR (600MHz, DMSO-d
6): δ ppm 4.71 (s, 2H ,-NC
h 2-); (7.04 t, 2H); (7.24 t, 1H); (7.33 t, 2H); (7.38 d, 2H); (7.68 s, 2H); (8.14 s, 1H); (8.32 s, 1H); (9.16 s, 1H); (12.93 s, 1H)
(9) 4-((6-benzylamine-9H-purine-2)-amido) benzsulfamide (4g)
By compound 3 (0.6g, 2.47mmol) add in 10mL trifluoroethanol (TFE), open induction stirring, then add sulfanilamide (SN) (1.7g, 9.87mmol) and trifluoroacetic acid (1.83mL, 24.7mmol), mixed solution is as in 85 ℃ of oil baths, prolong cooling for reflux.TLC monitoring, treats compound 3 after completion of the reaction, stops refluxing, reaction solution is cooling, concentrated, in resistates, add after methyl alcohol, be stirred to abundant dissolving, obtain white suspension, filter, collect filtrate, the crude product obtaining obtains white solid 0.07g (0.18mmol through purification by silica gel column chromatography, 8%), mp 215-218 ℃ of .HRMS:m/z calcd for C
18h
17n
7o
2s[M+1]
+396.1242Found:396.1238;
1h-NMR (600MHz, DMSO-d
6): δ ppm 4.72 (s, 2H); (7.10 s, 2H); (7.22 s, 1H); (7.32 s, 2H); (7.39 d, 2H); (7.60 d, 2H); (7.86 s, 2H); (7.90 s, 1H); (8.19 s, 1H); (9.32 s, 1H); (12.55 s, 1H)
(10) N
6-benzyl-N
2-phenyl-9H-purine-2,6-diamines (4h)
The same 4a of method.Compound 3 (0.47g, 1.9mmol), aniline (1.3ml, 13.2mmol), trifluoroacetic acid (0.1ml, 1.3mmol), n-butanol (10ml).Brown color solid 0.5g (1.6mmol, 83%), mp 261-262 ℃ of .HRMS:m/z calcd forC
18h
16n
6[M+1]
+317.1514Found:317.1521;
1h-NMR (600MHz, DMSO-d
6): δ ppm 4.74 (s, 2H); (6.88 t, 1H); (7.20 t, 2H); (7.25 d, 1H); (7.33 t, 2H); (7.39 d, 2H); (7.70 d, 2H); (8.11 s, 1H); (8.32 s, 1H); (9.10 s, 1H); (12.81 s, 1H)
(11) N
6-benzyl-N
2-(4-first sulfo group phenyl)-9H-purine-2,6-diamines (4i)
The same 4g of method.Compound 3 (0.6g, 2.47mmol), to methylsulphonic acid aniline (1.69g, 9.87mmol), trifluoroacetic acid (1.83mL, 24.7mmol), trifluoroethanol (10mL).Off-white color solid 0.04g (0.1mmol, 4%), mp 241-244 ℃ of .HRMS:m/z calcd for C
18h
17n
7o
2s[M+1]
+395.1290Found:395.1305;
1h-NMR (600MHz, DMSO-d
6): δ ppm 3.14 (s, 3H); (4.78 s, 2H); (7.25 s, 1H); (7.35 s, 2H); (7.42 s, 2H); (7.73 d, 2H); (7.94 s, 2H); (8.52 s, 1H); (8.74 s, 1H); (9.81 s, 1H); (13.27 s, 1H)
The preparation of embodiment 2, compound 4j-4q
(1) N-(cyclohexyl methyl)-2-chloro-9H-purine-6-amine (6)
By 2,6-dichloropurine (5.0g, 26.5mmol) and propyl carbinol (30mL) add in eggplant-shape bottle, then under agitation condition, add successively aminomethyl hexanaphthene (4.2mL, 31.7mmol) and triethylamine (5.4mL, 40mmol), finish, mixed solution is placed in 130 ℃ of oil baths, prolong cooling for reflux.After TLC monitoring raw material total overall reaction, stop refluxing, reaction solution is cooling, separates out a large amount of white solids, suction filtration, after solid fully washs with n-propyl alcohol, collects, after vacuum-drying, obtain white solid 6.41g (24.1mmol, 91%), mp 208-210 ℃ of .HRMS:m/z calcd for C
12h
16clN
5[M+1]
+266.1172Found:266.1177;
1h-NMR (400MHz, DMSO-d
6): δ ppm 0.92-1.00 (m, 2H); 1.57-1.21 (m, 3H); 1.61-1.73 (m, 6H); 3.23 (s, 2H); 7.94 (s, 1H); 8.08 (s, 1H); 12.85 (s, 1H)
(2) 4-((6-cyclohexyl methylamine-9H-purine-2)-amido) benzsulfamide (4j)
Compound (6,0.6g, 2.26mmol) is added in 15mL trifluoroethanol, open induction stirring, then add sulfanilamide (SN) (1.56g, 9mmol) and trifluoroacetic acid (1.7mL, 22.6mmol), mixed solution is as in 90 ℃ of oil baths, prolong cooling for reflux.TLC monitoring, treats compound 6 after completion of the reaction, stops refluxing, and reaction solution is cooling, evaporated under reduced pressure solvent, and residue obtains white solid 0.1g (0.25mmol, 11%), 258 ℃ of decomposition of mp after purification by silica gel column chromatography.HRMS:m/z?calcd?for?C
18H
23N
7O
2S[M+1]
+402.1712Found:402.1688;
1H-NMR(400MHz,DMSO-d
6):δppm?1.02(t,2H);1.19(t,3H);1.64(s,1H);1.70(s,3H);1.82(d,2H);3.39(s,2H);7.16(s,2H);7.70(d,2H);7.96(d,J＝8.4Hz,2H);8.22(s,1H);8.39(s,1H);9.65(s,1H);13.32(s,1H)
(3) N
6-cyclohexyl methyl-N
2-phenyl-9H-purine-2,6-diamines (4k)
Compound (6,0.6g, 2.25mmol) is added in propyl carbinol (15mL), open induction stirring, then add after aniline (1.5mL, 15.8mmol) and trifluoroacetic acid (0.1mL), mixed solution is placed in 130 ℃ of oil baths, prolong cooling for reflux.TLC monitoring, treats compound 6 after completion of the reaction, stops refluxing, and reaction solution is cooling, separates out a large amount of solids.Suction filtration, methylene dichloride or methanol wash for solid, obtain white solid 0.47g (1.5mmol, 65%), mp>290 ℃ of .HRMS:m/z calcd for C after being dried
18h
22n
6[M+1]
+323.1984Found:323.1987;
1h-NMR (400MHz, DMSO-d
6): δ ppm 0.95-1.03 (m, 2H); 1.13-1.21 (m, 3H); 1.63-1.80 (m, 6H); 3.36 (s, 2H); 6.94 (t, 1H); 7.26 (t, 2H); 7.75 (d, 2H); 8.20 (s, 2H); 9.27 (s, 1H); 13.13 (s, 1H).
(4) 4-(6-cyclohexyl methylamine)-9H-purine-2-amido) phenol (4l)
The same 4k of method.Compound (6,0.6g, 2.25mmol), p-aminophenol (1.6g, 15.8mmol), trifluoroacetic acid (0.1mL), propyl carbinol (15mL).Off-white color solid 0.39g (1.2mmol, 51%), 285 ℃ of decomposition.HRMS:m/z?calcd?forC
18H
22N
6O[M+1]
+339.1933Found:339.1936;
1H-NMR(400MHz,DMSO-d
6):δppm?0.96(t,2H);1.17(t,3H);1.63-1.77(m,6H);3.31(s,2H);6.66(d,2H);7.52(d,2H);7.63(br?s,1H);7.84(s,1H);8.60(s,1H);8.90(s,1H);12.48(s,1H)
(5) N
6-cyclohexyl methyl-N
2-(4-fluorophenyl)-9H-purine-2,6-diamines (4m)
The same 4k of method.Compound (6,0.6g, 2.25mmol), para-fluoroaniline (1.5mL, 15.8mmol), trifluoroacetic acid (0.1mL), propyl carbinol (15mL).White solid 0.50g (1.5mmol, 65%), mp>300 ℃ of .HRMS:m/z calcd for C
18h
21fN
6[M+1]
+341.1890Found:341.1876;
1h-NMR (400MHz, DMSO-d
6): δ ppm 0.93-1.02 (m, 2H); 1.13-1.24 (m, 3H); 1.63-1.78 (m, 6H); 3.34 (s, 2H); 7.10 (t, 2H); 7.73-7.77 (m, 2H); 8.23 (s, 2H); 9.30 (s, 1H); 12.32 (s, 1H)
(6) N
6-cyclohexyl methyl-N
2-(4-tolyl)-9H-purine-2,6-diamines (4n)
The same 4k of method.Compound (6,0.6g, 2.25mmol), to monomethylaniline (1.69g, 15.8mmol), trifluoroacetic acid (0.1mL), propyl carbinol (15mL).White solid 0.45g (1.3mmol, 60%), mp>300 ℃ of .HRMS:m/z calcd for C
19h
24n
6[M+1]
+337.2140Found:337.2139;
1h-NMR (400MHz, DMSO-d
6): δ ppm 0.99 (t, 2H); 1.13-1.21 (m, 3H); 1.63-1.78 (m, 6H); 2.25 (s, 3H); 3.36 (s, 2H); 7.07 (d, 2H); 7.62 (d, 2H); 8.15 (s, 1H); 8.24 (s, 1H); 9.20 (s, 1H); 13.13 (s, 1H)
(7) N
6-cyclohexyl methyl-N
2-(4-p-methoxy-phenyl)-9H-purine-2,6-diamines (4o)
The same 4k of method.Compound (6,0.6g, 2.25mmol), P-nethoxyaniline (1.95g, 15.8mmol), trifluoroacetic acid (0.1mL), propyl carbinol (15mL).White solid 0.48g (1.4mmol, 61%), mp>300 ℃ of .HRMS:m/z calcd forC
19h
24n
6o[M+1]
+353.2090Found:353.2078;
1h-NMR (400MHz, DMSO-d
6): δ ppm 0.93-1.02 (m, 2H); 1.13-1.24 (m, 3H); 1.63-1.77 (m, 6H); 3.35 (s, 2H); 3.73 (s, 3H); 6.86 (d, 2H); 7.60 (d, 2H); 8.08 (s, 1H); 8.17 (s, 1H); 9.06 (s, 1H); 13.01 (s, 1H)
(8) N
6-cyclohexyl methyl-N
2-(4-bromophenyl)-9H-purine-2,6-diamines (4p)
The same 4k of method.Compound (6,0.6g, 2.25mmol), para-bromoaniline (2.72g, 15.8mmol), trifluoroacetic acid (0.1mL), propyl carbinol (15mL).White solid 0.53g (1.3mmol, 59%), mp>300 ℃ of .HRMS:m/z calcd for C
18h
21brN
6[M+1]
+401.1089Found:401.1114;
1h-NMR (400MHz, DMSO-d
6): δ ppm 1.02 (d, 2H); (1.18 t, 3H); 1.63-1.79 (m, 6H); 3.36 (s, 2H); 7.42 (d, 2H); 7.76 (d, 2H); 8.20 (s, 1H); 8.32 (s, 1H); 9.41 (s, 1H); 13.35 (s, 1H)
(9) N
6-cyclohexyl methyl-N
2-(4-first sulfo group phenyl)-9H-purine-2,6-diamines (4q)
The same 4j of method.Compound (6,0.6g, 2.26mmol), to methylsulphonic acid aniline (1.55g, 9mmol), trifluoroacetic acid (1.7mL, 22.6mmol), trifluoroethanol (12mL).White solid 0.05g (0.12mmol, 6%), mp 250-253 ℃ of .HRMS:m/zcalcd for C
19h
24n
6o
2s[M+1]
+401.1759Found:401.1762;
1h-NMR (400MHz, DMSO-d
6): δ ppm1.03 (d, 2H); 1.17-1.22 (m, 3H); 1.63-1.83 (m, 6H); 3.14 (s, 3H); 3.41 (s, 2H); 7.78 (d, 2H); 8.05 (d, 2H); 8.34 (s, 1H); 8.41 (s, 1H); 9.81 (s, 1H); 13.16 (s, 1H)
The preparation of embodiment 3, compound 10a-10i
(1) N-(cyclohexyl methyl)-2-fluoro-9H-purine-6-amine (8)
By fluoro-2-6-chloropurine (5.0g, 29mmol) be dissolved in dehydrated alcohol (30mL), then add successively aminomethyl hexanaphthene (4.5mL, 34.9mmol) and triethylamine (5.9mL, 43.6mmol), finish, mixed solution is placed in 80 ℃ of oil baths, prolong cooling for reflux.After TLC monitoring raw material total overall reaction, stop refluxing, cooling, after reaction solution evaporated under reduced pressure, obtain light yellow solid, after vacuum-drying, after column chromatography purification, obtain white solid powder 4.80g (19.3mmol, 66%), mp 164-168 ℃ of .ESI-MS:m/z 250.4[M+1]
+;
1h-NMR (300MHz, DMSO-d
6): δ ppm 0.88-0.99 (m, 2H); 1.14-1.16 (t, 3H); 1.61-1.72 (m, 6H); 3.25 (s, 2H); 8.06 (s, 1H); 8.17 (s, 1H); 12.98 (s, 1H)
(2) N-(cyclohexyl methyl)-2-fluoro-9-sec.-propyl-9H-purine-6-amine (9)
By compound (8,4.80g, 19.3mmol) be dissolved in dimethyl sulfoxide (DMSO) (15mL), then add successively bromo propane (4.5mL, 48.1mmol) and Anhydrous potassium carbonate (8.0g, 57.8mmol), finish, mixed solution is placed in 80 ℃ of reactions of oil bath.TLC monitoring, after treating raw material total overall reaction, reaction solution dilute with water, is transferred in separating funnel, is extracted with ethyl acetate, and merges organic layer, then with saturated sodium-chloride washing, divides and get organic layer, anhydrous magnesium sulfate drying.Filter, decompression and solvent recovery, crude product obtains white solid powder 4.03g (13.8mmol, 72%), mp 112-115 ℃ .ESI-MS:m/z292.4[M+1 after purification by silica gel column chromatography]
+;
1h-NMR (400MHz, DMSO-d
6): δ ppm 0.94 (m, 2H); 1.14-1.16 (m, 3H); 1.49 (d, 6H); 1.64-1.67 (m, 6H); 3.26 (t, 2H); 4.59-4.65 (m, 1H); 8.20 (s, 1H); 8.29 (s, 1H)
(3) 2-(6-(cyclohexyl methylamine base)-9-sec.-propyl-9H-purine-2-amido)-n-butyl alcohol (10a)
By compound (9,0.6g, 2.1mmol), 2-amino-n-butyl alcohol (1.2mL, 13.4mmol) adds in dimethyl sulfoxide (DMSO) (10mL), and under nitrogen protection, after extracting vacuum, mixed solution is placed in 130 ℃ of reactions of oil bath.TLC monitoring, treats compound 9 after completion of the reaction, and stopped reaction, after reaction solution thin up, is extracted with ethyl acetate, and combined ethyl acetate layer, with saturated sodium-chloride washing, anhydrous magnesium sulfate drying.Filter, decompression and solvent recovery, crude product obtains .mp 116-118 ℃ of .HRMS:m/z calcd for C of white solid 0.23g (0.6mmol, 32.3%) through column chromatography purification
19h
32n
6o[M+1]
+361.2716Found:361.2711;
1h-NMR (600MHz, CDCl
3): 0.97-1.01 (m, 2H); (1.03 t, 3H); 1.13-1.26 (m, 3H); 1.53 (d, 6H); 1.56-1.60 (m, 2H); 1.61-1.67 (m, 2H); 1.71-1.73 (m, 2H); 1.82 (d, 2H); 3.38 (s, 2H); 3.62-3.65 (dd, 1H); 3.82-3.89 (m, 2H); 4.57-4.61 (m, 1H); 4.88 (s, 1H); 5.45-5.67 (d, 2H); 7.49 (s, 1H).
(4) 2-(6-(cyclohexyl methylamine base)-9-sec.-propyl-9H-purine-2-amido) ethanol (10b)
The same 10a of method.Compound (9,0.5g, 1.7mmol), thanomin (1mL, 11.2mmol), DMSO (10mL)..mp 95-98 ℃ of .HRMS:m/z calcd for C of off-white color solid 0.18g (0.5mmol, 31.6%)
17h
28n
6o[M+1]
+333.2401Found:333.2418;
1h-NMR (600MHz, CDCl
3): 0.94-1.03 (m, 2H); 1.18-1.27 (m, 3H) 1.53 (d, 6H); 1.55-1.61 (m, 1H); 1.63 (t, 1H), 1.72 (t, 2H); 1.81 (d, 2H); 3.38 (s, 2H); 3.55-3.58 (m, 2H); 3.83 (t, 2H); 4.57-4.64 (m, 1H); (5.02 br s, 1H), 5.37 (t, 1H); 5.87 (s, 1H); 7.50 (s, 1H)
(5) 2-(6-(cyclohexyl methylamine base)-9-sec.-propyl-9H-purine-2-amido)-3-hydroxy-propionic acid (10c)
By compound (9; 0.5g; 1.7mmol); Serine (3.6g; 34mmol), with 1,8-diazabicylo [5.4.0] 11 carbon-7-alkene (DBU) (5.3mL, 34mmol) adds in N-Methyl pyrrolidone (10mL); under nitrogen protection, after extracting vacuum, mixed solution is placed in 160 ℃ of reactions of oil bath.TLC monitoring, treats compound 9 after completion of the reaction, stopped reaction, and reaction solution dilutes with 10% citric acid, dichloromethane extraction, combined dichloromethane layer, with saturated sodium-chloride washing, anhydrous magnesium sulfate drying.Filter, decompression and solvent recovery, crude product obtains .mp 158-160 ℃ of .HRMS:m/zcalcd for C of off-white color solid 0.11g (0.3mmol, 17%) after column chromatography purification
18h
28n
6o
3[M+1]
+377.2301Found:377.2319;
1h-NMR (300MHz, CD
3oD): 0.98-1.12 (m, 2H); 1.29-1.40 (m, 3H); 1.58 (d, 6H); 1.65-1.73 (m, 2H); 1.78-1.89 (m, 4H); 3.38 (s, 2H); 4.03 (d, 2H); 4.61 (t, 1H); 4.66-4.72 (m, 1H); 7.86 (s, 1H).
(6) 2-(6-(cyclohexyl methylamine base)-9-sec.-propyl-9H-purine-2-amido)-3-hydroxybutyrate (10d)
The same 10c of method.Compound (9,0.6g, 2.1mmol), L-threonine (5.0g, 41.2mmol), DBU (6.3mL, 41.2mmol), N-Methyl pyrrolidone (10mL)..mp 138-142 ℃ of .HRMS:m/zcalcd for C of off-white color solid 0.15g (0.38mmol, 18.8%)
19h
30n
6o
3[M+1]
+391.2457Found:391.2469;
1h-NMR (300MHz, CD
3oD): 0.95-1.07 (m, 2H); 1.17-1.27 (m, 3H); 1.30 (d, 3H); 1.53 (d, 6H); 1.58-1.83 (m, 6H); 3.34 (s, 2H); 4.22-4.37 (m, 1H); 4.51-4.57 (m, 1H); 4.58-4.68 (m, 1H); 7.81 (s, 1H)
(7) 2-(6-(cyclohexyl methylamine base)-9-sec.-propyl-9H-purine-2-amino) propane-1,3-glycol (10e)
The same 10a of method.Compound (9,0.3g, 1mmol), L-serinol (0.61g, 6.7mmol), dimethyl sulfoxide (DMSO) (4mL)..mp 166-168 ℃ of .HRMS:m/z calcd for C of white solid 0.15g (0.4mmol, 40.5%)
18h
30n
6o
2[M+1]
+363.2508Found:363.2536;
1h-NMR (600MHz, CDCl
3): 0.93-0.99 (m, 2H); 1.09-1.27 (m, 3H); 1.50 (d, 6H); 1.53-1.60 (m, 1H); 1.63 (d, 1H), 1.69 (d, 2H); 1.78 (d, 2H); 3.32 (s, 2H); 3.82-3.90 (m, 4H); 4.08 (t, 1H); 4.53-4.59 (hept., 1H); 5.12 (s, 2H); 5.70 (d, 1H); 5.96 (s, 1H); 7.50 (s, 1H)
(8) 2-(6-(cyclohexyl methylamine base)-9-sec.-propyl-9H-purine-2-amido)-4-methylpentane-1-alcohol (10f)
The same 10a of method.Compound (9,0.3g, 1mmol), L-leucinol (0.86g, 6.7mmol), diisopropylethylamine (1.7mL, 10mmol), dimethyl sulfoxide (DMSO) (4mL).Faint yellow gluey liquid, with after the saturated ethyl acetate salify of hydrogenchloride .mp 145-149 ℃ of .HRMS:m/z calcd for C of white solid 0.08g (0.19mmol, 18.2%)
21h
36n
6o[M+1]
+389.3029Found:389.3007;
1h-NMR (400MHz, CD
3oD): 1.02 (t, 6H); 1.07-1.15 (m, 2H); 1.25-1.36 (m, 3H); 1.51-1.59 (m, 2H); 1.62 (t, 6H); 1.73-1.81 (m, 4H); 1.92 (d, 2H); 3.43-3.47 (m, 2H); 3.64-3.72 (m, 2H); 4.24-4.27 (m, 1H); 4.74-4.80 (m, 1H); 8.51 (s, 1H)
(9) 2-(6-(cyclohexyl methylamine base)-9-sec.-propyl-9H-purine-2-amido)-3-methyl-1-butanol (10g)
The same 10a of method.Compound (9,0.6g, 2.1mmol), L-valerian ammonia alcohol (1.5mL, 13.4mmol), diisopropylethylamine (3.4mL, 20.6mmol), dimethyl sulfoxide (DMSO) (6mL).Faint yellow gluey liquid, with after the saturated ethyl acetate salify of hydrogenchloride 75-79 ℃ of .HRMS:m/z calcd for C of off-white color solid 0.15g (0.3mmol.17.6%) .mp
20h
34n
6o[M+1]
+375.2872Found:375.2884;
1h-NMR (400MHz, CD
3oD): 1.03-1.07 (m, 6H); (1.13 d, 2H); 1.24-1.35 (m, 3H); 1.58-1.61 (dd, 6H); 1.72-1.81 (m, 4H); 1.89 (d, 2H); 2.07 (t, 1H); 3.41 (s, 2H); 3.75 (d, 2H); 4.02 (s, 1H); 4.67-4.73 (m, 1H); 8.04 (s, 1H)
(10) 2-(6-(cyclohexyl methylamine base)-9-sec.-propyl-9H-purine-2-amido) propane-1-alcohol (10h)
The same 10a of method.Compound (9,0.6g, 2.1mmol), L-aminopropanol (1.0g, 13.4mmol), diisopropylethylamine (3.4mL, 20.6mmol), dimethyl sulfoxide (DMSO) (8mL).Faint yellow gluey liquid, with after the saturated ethyl acetate salify of hydrogenchloride .mp 184-188 ℃ of .HRMS:m/z calcd for C of white solid 0.55g (1.4mmol, 70.5%)
18h
30n
6o[M+1]
+347.2559Found:347.2556;
1h-NMR (400MHz, CD
3oD): 1.09-1.17 (m, 2H); 1.24-1.30 (m, 3H); 1.34 (d, 3H); 1.67 (d, 6H); 1.74 (d, 2H); 1.81 (d, 2H); 1.95 (d, 2H); 3.52 (d, 2H); 3.63-3.74 (m, 2H); 4.20-4.24 (m, 1H); 4.84-4.90 (m, 1H); 9.00 (s, 1H)
(11) 2-(6-(cyclohexyl methylamine base)-9-sec.-propyl-9H-purine-2-amido) propionic acid (10i)
The same 10c of method.Compound (9,0.6g, 2.1mmol), ALANINE (3.7g, 41.2mmol), DBU (6.3mL, 41.2mmol), N-Methyl pyrrolidone (4mL)..mp 84-90 ℃ of .HRMS:m/z calcd for C of off-white color solid 0.21g (0.6mmol, 28.4%)
18h
28n
6o
2[M+1]
+361.2352Found:361.2354;
1h-NMR (400MHz, CD
3oD): 0.98-1.07 (m, 2H); 1.19-1.33 (m, 3H); 1.51 (d, 3H); 1.55-1.57 (dd, 6H); 1.69 (d, 2H); 1.77 (d, 2H); 1.85 (d, 2H); 3.38 (s, 2H); 4.43-4.48 (m, 1H); 4.61-4.68 (m, 1H); 7.82 (s, 1H)
Embodiment 4, target compound suppress the preliminary activity experiment (In vitro) of cell cycle protein dependent kinase
1) material: CDK1/cyclin B Kinase (CDK1/cyclin B kinases); Kinase-Glo Plus reagent (Kinase-
plus reagent); ATP (Triphosaden);
Substrate (substrate): Histone H derived peptide (Histone H derived peptide, Histidine H derived peptide): H09-19T;
Assay Buffer:25mM HEPES (4-hydroxyethyl piperazine ethanesulfonic acid), 10mM MgCl
2, 0.01%Triton X-100,100 μ g/mL BSA, 2.5mM DTT (dithiothreitol (DTT)), pH7.4.
Roscovitine: cell cycle protein dependent kinase inhibitor series antineoplastic medicament, is now just carrying out the clinical II phase and is studying.
2) method: the solution that compound is mixed with to 10 μ g/mL.In 384 orifice plates the every hole of experimental port add 1 μ L measure concentration be the testing compound of 10 μ M, blank well (ZPE) and negative control hole (HPE) add 1 μ L 10%DMSO solution.Then the every hole of testing compound experimental port and ZPE adds 4 μ L 2.5 × CDK1/cyclin B kinases, adds 4 μ L assaybuffer in HPE hole.Mixed solution mixes rear every hole and adds 5 μ L 2 × ATP-substrate mixtures (ATP and substrate mixed solution), and mixed solution mixes, and under 30 ℃ of conditions, hatches 1 hour.Every hole adds 10 μ L Kinase Glo Plus reagent, and mixed solution mixes under 27 ℃ of conditions and hatches 20min.Finally on EnVision, read luminous intensity, calculate inhibiting rate.Experimental result in Table 4, table 5.
The external enzyme experimental result that presses down of table 4. target compound 4a-4q
The external enzyme experimental result that presses down of table 5. target compound 10a-10i
By CDK1/cyclin B kinases tentatively being suppressed to activity experiment, find, compound 4g, 4j, 4l, 4m, 4q, 10a are better than positive control drug Roscovitine in the inhibition activity of 10 μ M concentration levels, and compound 4e, 4i, 4o, 10g and 10h are more approaching with the inhibition specific activity of Roscovitine on 10 μ M concentration levels simultaneously.
Embodiment 5, target compound suppress the activity test (In vitro) of cell proliferation
Target compound is carried out to the activity experiment of vitro inhibition cancer cell multiplication, the results are shown in Table 6, table 7.
1) material: MD-MBA-231 cell strain, the blue MTT of tetramethyl-azo azoles, 10% foetal calf serum, 96 orifice plates.
2) method:
Cell cultures: MD-MBA-231 tumor cell line all adopts cellar culture.During experiment, all use logarithmic phase cell.
Growth of Cells detects (mtt assay): MD-MBA-231 cell suspension is all adjusted to 5 × 10
4/ mL, is inoculated in respectively 96 orifice plates (100 μ L/ hole), 5000 cells/well.After bed board 4h, in every hole, add the substratum of 100 μ L containing different concns compound, compound final concentration in hole is respectively: 100,50,25,12.5,6.25 μ g/mL, each concentration is established four multiple holes, while not adding the hole reading of cell, make blank well, add the hole that cell do not add compound and make negative control hole, Roscovitine makes compound positive control.In 37 ℃, 5%CO
2in hatch 48h, every hole adds the MTT staining fluid of 10 μ L0.5%, continue to hatch after 4h, 2500rpm, centrifugal 12min, then abandons substratum in plate hole, adds DMSO solution, 100 μ L/ holes.The absorbancy OD value of measuring every hole in microplate reader in 570nm place, inhibitory rate of cell growth is calculated as follows:
The antiproliferative experimental result of table 6. target compound 4a-4q
ain table, numerical value is the mean value of three tests, the numeric representation standard deviation after " ± ".
The antiproliferative experimental result of table 7. target compound 10a-10i
ain table, numerical value is the mean value of three tests, the numeric representation standard deviation after " ± ".
Upper table test data shows, compound 4a, 4e, 4g, 4h, 4i, 4j, 4k, 4l, 4m, 4o, 4p, 4q, 10a, 10f, 10g and 10h will get well MD-MBA-231 cell inhibitory effect specific activity positive control Roscovitine in vitro.
Claims (6)
1.2-amino-6-aminomethyl purine compound or its pharmacy acceptable salt, steric isomer, it is characterized in that one of following compound:
N
6-benzyl-N
2-(4-bromo phenyl)-9H-purine-2,6-diamines (4a),
N
6-benzyl-N
2-(4-p-methoxy-phenyl)-9H-purine-2,6-diamines (4b),
N
6-benzyl-N
2-p-methylphenyl-9H-purine-2,6-diamines (4c),
N
6-benzyl-N
2-(3-methoxyphenyl)-9H-purine-2,6-diamines (4d),
4-((6-benzylamine-9H-purine-2)-amido) phenol (4e),
N
6-benzyl-N
2-(4-fluorophenyl)-9H-purine-2,6-diamines (4f),
4-((6-benzylamine-9H-purine-2)-amido) benzsulfamide (4g),
N
6-benzyl-N
2-phenyl-9H-purine-2,6-diamines (4h),
N
6-benzyl-N
2-(4-first sulfo group phenyl)-9H-purine-2,6-diamines (4i),
4-((6-cyclohexyl methylamine-9H-purine-2)-amido) benzsulfamide (4j),
N
6-cyclohexyl methyl-N
2-phenyl-9H-purine-2,6-diamines (4k),
4-(6-cyclohexyl methylamine)-9H-purine-2-amido) phenol (4l),
N
6-cyclohexyl methyl-N
2-(4-fluorophenyl)-9H-purine-2,6-diamines (4m),
N
6-cyclohexyl methyl-N
2-(4-tolyl)-9H-purine-2,6-diamines (4n),
N
6-cyclohexyl methyl-N
2-(4-p-methoxy-phenyl)-9H-purine-2,6-diamines (4o),
N
6-cyclohexyl methyl-N
2-(4-bromophenyl)-9H-purine-2,6-diamines (4p),
N
6-cyclohexyl methyl-N
2-(4-first sulfo group phenyl)-9H-purine-2,6-diamines (4q).
2. the preparation method of compound described in claim 1, is characterized in that the preparation process of compound 4a-4i is as follows:
Take 2-amido-6-chloropurine as raw material, obtain 2-amino-6-substituted purin with benzylamine generation nucleophilic substitution reaction, then at HBF
4and NaNO
2under effect, through Ba Erci-Xi Man (Balz-Schiemann) reaction, make the fluoro-6-substituted purin of 2-, finally generate target compound 4a-4i with the aniline reaction of various replacements again; Synthetic route one is as follows:
Wherein, R
1for phenyl, and R
2for there is no the phenyl of the following group replacement of replacement or quilt: 4-Br, 4-OCH
3, 4-CH3,3-OCH
3, 4-OH, 4-F, 4-SO
2hN
2, 4-SO
2cH
3;
Reagent and condition in said synthesis route one reaction formula: a. propyl carbinol, triethylamine, 130 ℃ of backflows; B.HBF
4, NaNO
2, H
2o ,-15 ℃; C. trifluoroacetic acid (TFA), propyl carbinol, 130 ℃ or trifluoroacetic acid (TFA), trifluoroethanol (TFE), 85 ℃.
3. the preparation method of compound described in claim 1, is characterized in that the preparation process of compound 4j-4q is as follows:
With 2,6-dichloropurine, for raw material, and after cyclohexyl methyl amine generation nucleophilic substitution reaction, obtain the chloro-6-substituted purin of 2-, and then generate target compound 4j-4q with the aniline reaction of various replacements; Synthetic route two is as follows:
Wherein, R
1for cyclohexyl, R
2for there is no the phenyl of the following group replacement of replacement or quilt: 4-Br, 4-OCH
3, 4-CH
3, 4-OH, 4-F, 4-SO
2hN
2, 4-SO
2cH
3;
Reagent and condition in said synthesis route two reaction formula: d. propyl carbinol, triethylamine, 130 ℃ of backflows; E. trifluoroacetic acid, propyl carbinol, 130 ℃ or trifluoroacetic acid, trifluoroethanol, 85 ℃.
4. the application of the compound described in claim 1 in the medicine of preparing the mammalian diseases that prevention or treatment are relevant to the active imbalance of cell cycle protein dependent kinase; The described related mammalian disease with the active unconventionality expression of cell cycle protein dependent kinase comprises cancer, neurodegenerative disease, malaria and diabetes.
5. be suitable for the oral mammiferous pharmaceutical composition that gives, comprise the compound described in claim 1 and one or more pharmaceutically acceptable carriers or vehicle.
6. be suitable for parenteral and give a mammiferous pharmaceutical composition, comprise the compound described in claim 1 and one or more pharmaceutically acceptable carriers or vehicle.
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| Medicinal Chemistry》.2011,第19卷6949-6965. * |
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