The preparation method of piperidines with antitumor activity and pyridine compounds and theirTechnical field
The invention belongs to pharmaceutical chemical synthesis technical field, and in particular to a kind of piperidines with antitumor activity and pyrroleThe preparation method of pyridine class compound.
Background technology
In recent years, heterocyclic compound is because structure is changeable, active high, toxicity is low and turns into the master of medicine and agricultural chemicals innovationStream, plurality of new varieties just continuously put goods on the market, and study on the synthesis is more towards baroque condensed hetero ring, double miscellaneousRing and poly-heterocyclic compounds.Piperidines and azole compounds are all the nitrogen-containing heterocycle compounds with good biological activity, extensively shouldFor medicine and the study on the synthesis of agricultural chemicals.Piperidines is mainly used in synthesis medicine, agricultural chemicals and rubber chemicals, is mainly used in pesticide industryIt is a kind of selective non-hormone-type thiocarbamic acid class herbicide, very with hair in synthesis herbicides for use in paddy dimepiperateExhibition prospect.It is used to synthesize medicine for digestive system hydrochloric acid acetyl Roxatidine, cardiovascular disease medicine Dipyridmole in pharmaceuticals industryDeng.It is used to synthesize the super vulcanization accelerator bis-pentamethylenethiuram tetrasulfide of thiurams in Rubber Chemicals Industries, two is thioSuper accelerator pentamethylene aminodithioformic acid piperidinium salt of Carbamates etc..Other piperidines can also synthesize a variety of newType fine-chemical intermediate, many products belong to the centre of small tonnage newly developed, the medicine of high added value, agricultural chemicals and auxiliary agentBody, such as pipecoline, 3- aminomethylpiperidines, 4- hydroxy piperidines.Pyridine is also a kind of important nitrogen heterocyclic ring, because it hasGood bioactivity is widely used in medical research.For example, its derivative can be used as 5HT2A receptor antagonists, cellExternal signal regulatory protein kinase inhibitor, mammal P2X7 conditioning agents, and increase with anti-breast cancer cell MDA-MB-231Grow activity and suppress the propagation of hepatocellular carcinoma H22.We have synthesized a series of with antitumor in laboratory by new methodThe piperidines and pyridine compounds and their of activity, and carried out corresponding bioactivity detection.
The content of the invention
Present invention solves the technical problem that there is provided, a kind of synthetic method is simple, and having for molecular structure novelty is antitumorThe piperidines and pyridine compounds and their preparation method of activity.
The present invention is to solve above-mentioned technical problem to adopt the following technical scheme that, a kind of new piperazine with antitumor activityPyridine and pyridine compounds and their preparation method, it is characterised in that its molecular structure is：R is benzene, to fluorineBenzene, p-nitrophenyl
The present invention is to solve above-mentioned technical problem to adopt the following technical scheme that, a kind of new piperazine with antitumor activityPyridine and pyridine compounds and their preparation method, it is characterised in that concretely comprise the following steps：
A, N-Boc-4- piperidones and dimethyl carbonate react in the presence of potassium tert-butoxide obtains N-Boc-3- formic acid firstEster -4- piperidones
B, N-Boc-3- methyl formate -4- piperidones are under ammonium acetate effect, and ketone carbonyl redox is changed into aminoCompound N-Boc-3- methyl formate -4- amino -3- alkene-piperidines
C, N-Boc-3- methyl formate -4- amino -3- alkene-piperidines and chloroformyl ethyl acetate take under TEA effectsGeneration reaction obtains compound N-Boc-3- methyl formate -4- carbamyl ethyl acetate -3- alkene-piperidines
D, N-Boc-3- methyl formate -4- carbamyl ethyl acetate -3- alkene-piperidines occur in the presence of potassium tert-butoxideIntramolecular cyclization obtains compoundThen the compound carries out intramolecular in acid conditionHydrogen migration and carbonyl reduction obtain compound
E、Under strongly acidic conditions, ester group and Boc groups are sloughed in heating, obtain compound
F、Under POCl3 effect, hydroxyl is replaced by chlorine obtains compound
G、Chloro carbon-to-nitrogen double bon is changed into amido link under concentrated hydrochloric acid effect and obtain compound
H、In the basic conditions, Boc amido protectings are carried out and obtains compound
I、Under cesium carbonate effect compound is obtained with iodomethane reaction
J、Reacted with NBS, bromination is carried out on methyl and obtains compound
K、Reacted with hydrazine hydrate and obtain compound
L、Slough progress molecular weight cyclization after a molecule HBr and obtain compound
M、Slough Boc groups and obtain compound
N、React and generate with m-trifluoromethyl phenyl boric acid under potassium phosphate effect
O、Obtained with carboxylic acid compound in alkalescence condition reaction
Further limit, step A detailed process is：In reaction bulb, 1eq N-Boc-4- piperidones is added toIn the toluene of 10V volumes, 2eq dimethyl carbonate and 2eq potassium tert-butoxide are added, 70 DEG C of reaction 1h is heated to, is cooled toRoom temperature, adds water and is quenched, and it is 7 to adjust reaction solution pH with 1mol/L HCl, ethyl acetate extraction, after anhydrous sodium sulfate drying, is spin-dried forObtain yellow oil N-Boc-3- methyl formate -4- piperidones
Further limit, step B detailed process is：1eq N-Boc-3- methyl formate -4- piperidones is added to 10In the methanol of times volume, 3eq ammonium acetate is added, reaction is stayed overnight, be spin-dried for methanol, add the water of 3 times of volumes, dichloromethane extractionAnhydrous sodium sulfate drying is used after extracting reaction solution, red oily liquids N-Boc-3- methyl formate -4- amino -3- is obtained after being spin-dried forAlkene-piperidines
Further limit, step C detailed process is：By 1eq N-Boc-3- methyl formate -4- amino -3- alkene-piperidinesIn the DCM for being added to 8 times of volumes, 1.05eq TEA is added, 10 DEG C are cooled to, 1.05eq 4- chloromethane ethyl acetoacetic acid second is added dropwiseEster, room temperature reaction is stayed overnight, and adds the DCM dilute reaction solutions of 8 times of volumes, twice, anhydrous sodium sulfate drying is spin-dried for both obtaining for washingRed oil product N-Boc-3- methyl formate -4- carbamyl ethyl acetate -3- alkene-piperidines
Further limit, step D detailed process is：1eq N-Boc-3- methyl formate -4- carbamyl acetic acid secondEster -3- alkene-piperidines is added in the THF of 10 times of volumes, then is added portionwise 2.0eq t-BuOK, reaction temperature control less than25 DEG C, frozen water is added after reaction 1h and is quenched, it is 3 to adjust reaction solution pH with 2mol/L HCl, filtering, and it is white that vacuum drying obtains classColor solid product
Further limit, step E detailed process is：In 10eq 6mol/L HCl solution, it is added portionwise 1.0eq's100 DEG C are heated to, reaction is stayed overnight, are spin-dried for reaction dissolvent, then washed with ether, is dried in vacuoObtain off-white powder
Further limit, step F detailed process is：To 5eq POCl3In be added portionwise 1.0eq's100 DEG C are heated to, reaction is stayed overnight, are spin-dried for POCl3Obtain Red oil product
Further limit, step G detailed process is：It is added to the 1,4- dioxies six of 4 times of volumesIn ring, the concentrated hydrochloric acid of 4 times of volumes is added, 100 DEG C are heated to, back flow reaction 2 days is spin-dried for adding the second of 10 times of volumes after solventAcetoacetic ester, is washed three times, is dried and is obtained brown solid after being spin-dried for
Further limit, step H detailed process is：1eq'sIt is added to the 1,4- of 10 times of volumesIn dioxane and the water of 10 times of volumes, then 3.0eq sodium carbonate and 1.5eq (Boc) is added portionwise2O, reacts at room temperature 10hAfterwards, filter, then wash with ethyl acetate reaction solution is extracted with ethyl acetate after filter cake, then washed with sodium chloride solution, dried, revolvedReaction solution is obtained
Further limit, step I detailed process is：1.0eq'sIt is added to 10 times of volumesDMF in, add 1.5eq Cs2CO3, 1.3eq KI, room temperature reaction stays overnight, and adds frozen water and reaction solution, acetic acid is quenchedEthyl ester extractive reaction liquid, sodium chloride solution washing, dries, is spin-dried for, then is beaten with ether, filters, and it is solid that vacuum drying obtains whiteBody
Further limit, step J detailed process is：In reaction bulb,It is added to NBSIn toluene, it is heated to after 80 DEG C, reaction a period of time, TLC monitoring raw material reactions are complete, and reaction solution is added in frozen water, there is bigMeasure solid to separate out, suction filtration obtains compound
Further limit, step K detailed process is：In reaction bulb,With it is a certain amount ofHydrazine hydrate is added in DMF, is heated under nitrogen protection after 60 DEG C, reaction a period of time, and TLC monitoring raw material reactions are complete,Reaction solution is added in frozen water, there are a large amount of solids to separate out, suction filtration obtains compound
Further limit, step L detailed process is：In reaction bulb,It is added toIn DMSO, under air atmosphere, be heated to 130 DEG C, after reaction a period of time, TLC monitoring raw material reactions are complete, reaction solution plusEnter in frozen water, there are a large amount of solids to separate out, suction filtration obtains compound
Further limit, step M detailed process is：1.0eqIt is added to 10 times of volumesIn the HCl/1 of methanol and the 12mol/L of 10 volumes, 4- dioxane, room temperature reaction is stayed overnight, and is spin-dried for, ether washing, is obtained
Further limit, step N detailed process is：By 1.0eq'sIt is added to 20 times of volumesIn THF, 3eq 1mol/L potassium phosphate and 1.2eq m-trifluoromethyl phenyl boric acid are added, 100 DEG C is heated to, reactedNight, ethyl acetate extraction is dried, is spin-dried for rear column chromatography for separation and obtains
Further limit, step O detailed process is：In reaction bulb,It is added in DMF,Triethylamine and carboxylic acid compound are added, 70 DEG C are heated to, reaction a period of time obtains compound
The synthetic route of the new piperidines with antitumor activity of the present invention and pyridine compounds and their is：
The present invention has synthesized a series of piperidines and pyridine compounds and their and has carried out antitumor activity test, it is found that such is changedCompound has preferable inhibitory activity to breast cancer cell MCF-7 and liver cancer HepG2
Embodiment
The above to the present invention is described in further details by the following examples, but this should not be interpreted as to thisThe scope for inventing above-mentioned theme is only limitted to following embodiment, and all technologies realized based on the above of the present invention belong to this hairBright scope.
Embodiment 1
In reaction bulb, N-Boc-4- piperidones 20g (0.1mol) is added in toluene 200mL, carbonic acid two is addedMethyl esters 18g (0.2mol) and potassium tert-butoxide 22g (0.2mol), is heated to 70 DEG C of reaction 1h, is cooled to room temperature, the 100mL that adds water quenchesGo out, it is 7 to adjust reaction solution pH with 1mol/L HCl, ethyl acetate extraction, after anhydrous sodium sulfate drying, be spin-dried for obtaining yellow oilShape thing N-Boc-3- methyl formate -4- piperidones 25g；1HNMR(400MHz,CD3Cl)δ:3.81 (s, 1H), 3.71 (d, J=8.4Hz, 1H), 3.68 (d, J=8.4Hz, 1H), 3.45 (s, 3H), 3.07-3.05 (m, 2H), 2.76-2.73 (m, 2H), 1.37(s,9H).MS-ESI(m/z):258.3[M+H+]。
Embodiment 2
In reaction bulb, N-Boc-3- methyl formate -4- piperidones 25g (0.1mol) are added in methanol 300mL, thenAmmonium acetate 22g (0.3mol) is added, reaction is stayed overnight, TLC monitoring raw material reactions are complete, are spin-dried for methanol, water 900mL added, with twoChloromethanes 300mL extractive reactions liquid three times, merges and anhydrous sodium sulfate drying is used after organic phase, red oily liquids is obtained after being spin-dried forN-Boc-3- methyl formate -4- amino -3- alkene-piperidines 25g；1HNMR(400MHz,CD3Cl)δ:8.56(s,2H),3.93(s,2H),3.77(s,3H),3.57-3.55(m,2H),2.16-2.13(m,2H),1.37(s,9H).MS-ESI(m/z):257.3[M+H+]。
Embodiment 3
In reaction bulb, N-Boc-3- methyl formate -4- amino -3- alkene-piperidines 26g (0.1mol) is added to dichloromethaneIn alkane 200mL, TEA11g (0.11mol) is added, 10 DEG C are cooled to, 4- chloroformyl ethyl acetate 16g is slowly added dropwise(0.105mol), room temperature reaction is stayed overnight, and TLC monitoring raw material reactions are complete, add dichloromethane 200mL dilute reaction solutions, waterWash twice, anhydrous sodium sulfate drying, be spin-dried for both obtaining Red oil product N-Boc-3- methyl formate -4- carbamyl acetic acid secondEster -3- alkene-piperidines 25g；1HNMR(400MHz,CD3Cl)δ:4.71(s,2H),3.93(s,2H),3.79(s,3H),3.57-3.55(m,2H),3.53(s,2H),2.16-2.13(m,2H),1.37(s,9H),1.29(s,3H).MS-ESI(m/z):371.4[M+H+]。
Embodiment 4
In reaction bulb, N-Boc-3- methyl formate -4- carbamyl ethyl acetate -3- alkene-piperidines 37g (0.1mol)It is added in THF400mL, then t-BuOK 23g (0.2mol) is added portionwise, reaction temperature control is reacted after 1h less than 25 DEG CAdd frozen water 300mL to be quenched, it is 3 to adjust reaction solution pH with 2mol/L HCl, and filtering, vacuum drying obtains off-white powder productionProduct32g；1H NMR(400MHz,DMSO-d6)δ:11.51(s,1H),5.35(s,1H),4.71 (s, 2H), 4.33 (d, J=4.0Hz, 2H), 3.66-3.62 (m, 2H), 3.25 (d, J=12.0Hz, 2H), 1.41-1.39(m,9H),1.33-1.32(m,3H)。
Embodiment 5
In reaction bulb, the HCl solution 200mL in 6mol/L is added, then be added portionwise34g (0.1mol), is heated to 100 DEG C, reaction is stayed overnight, and is spin-dried for reaction dissolvent, then is washed with ether, and vacuum drying obtains off-white colorSolid15g；1H NMR(400MHz,DMSO-d6)δ:11.47(s,1H),5.95(s,1H),5.41(s,1H), 3.86-3.85 (m, 2H), 3.71 (d, J=12.0Hz, 2H), 3.11-3.09 (m, 2H), 1.90 (s, 1H).
Embodiment 6
In closed reaction bulb, it is added portionwise into POCl3 50g (0.5mol)16g(0.1mol), is slowly heated to 100 DEG C, reaction is stayed overnight, and vacuum is spin-dried for POCl3 and obtains Red oil product16g；1H NMR(400MHz,DMSO-d6)δ:7.61 (s, 1H), 3.81 (s, 2H), 3.37 (d, J=12.0Hz,2H),3.13-3.12(m,2H),1.87(s,1H)。
Embodiment 7
In the reaction bulb with thermometer and stirring,20g is added to 1,4- dioxaneIn 100mL, concentrated hydrochloric acid 100mL is slow added into, 100 DEG C are heated to, back flow reaction 2 days, TLC monitoring raw material reactions are complete, rotationEthyl acetate 200mL is added after dry solvent, is washed with water 100mL three times, organic phase obtains brown and consolidated after being spin-dried for after drying through vacuumBody17g；1H NMR(400MHz,CDCl3)δ:8.17(s,1H),6.62(s,1H),3.35(s,2H),2.96(d, J=12.0Hz, 2H), 2.07-2.05 (m, 2H) .MS-ESI (m/z):185.6[M+H+]。
Embodiment 8
In reaction bulb,18g (0.1mol) is added to 1,4- dioxane 200mL and water 200mLIn, then sodium carbonate 30g (0.3mol) and (Boc) is added portionwise2After O 33g (0.15mol), room temperature reaction 10h, TLC monitoring is formerMaterial reaction is complete, filtering reacting liquid, then is washed with ethyl acetate 100mL after filter cake with ethyl acetate 200mL extractive reactions liquid threeIt is secondary, then washed with sodium chloride solution, dry, rotation reaction solution is obtained20g
Embodiment 9
In reaction solution,28g (0.1mol) is added in DMF 300mL, adds carbonic acidCaesium 50g (0.15mol), KI 20g (0.13mol), room temperature reaction is stayed overnight, and TLC monitoring raw material reactions are complete, add frozen waterReaction solution is quenched in 100mL, and ethyl acetate 200mL extractive reactions liquid three times, saturated nacl aqueous solution 200mL washing reaction liquids are doneIt is dry, it is spin-dried for, then be beaten with ether, filter, vacuum drying obtains white solid26g；1H NMR(400MHz,DMSO-d6)δ:6.61 (s, 1H), 3.94 (s, 2H), 3.54 (s, 3H), 3.55 (d, J=8.0Hz, 2H), 2.07-2.05(m,2H),1.38(s,9H).MS-ESI(m/z):299.8[M+H+]。
Embodiment 10
In reaction bulb,30g (0.1mol) and NBS 20g are added in toluene 200mL, plusHeat is to 80 DEG C, after reaction a period of time, and TLC monitoring raw material reactions are complete, and reaction solution is added in frozen water, has a large amount of solids to analyseGo out, suction filtration obtains compound27g；1H NMR(400MHz,DMSO-d6)δ:6.63(s,1H),3.95(s, 2H), 3.51 (s, 2H), 4.97 (d, J=8.0Hz, 2H), 2.07-2.05 (m, 2H), 1.38 (s, 9H) .MS-ESI (m/z):378.6[M+H+]。
Embodiment 11
In reaction bulb,38g (0.1mol) and hydrazine hydrate 30mL are added to DMF 250mLIn, it is heated under nitrogen protection after 60 DEG C, reaction a period of time, TLC monitoring raw material reactions are complete, and reaction solution is added frozen waterIn, there are a large amount of solids to separate out, suction filtration obtains compound35g；1H NMR(400MHz,DMSO-d6)δ:7.07 (s, 2H), 6.25 (s, 1H), 4.97 (d, J=8.0Hz, 2H), 3.95 (s, 2H), 3.52 (s, 2H), 2.02-2.01 (m,2H),1.37(s,9H).MS-ESI(m/z):392.7[M+H+]。
Embodiment 12
In reaction bulb,39g is added in DMSO200mL, under air atmosphere, heatingTo 130 DEG C, after reaction a period of time, TLC monitoring raw material reactions are complete, and reaction solution is added in frozen water, has a large amount of solids to separate out,Suction filtration obtains compound31g；1H NMR(400MHz,DMSO-d6)δ:7.02(s,1H),6.26(s,1H), (s, the 9H) .MS- of 4.01 (d, J=8.0Hz, 2H), 3.95 (s, 2H), 3.52 (s, 2H), 2.02-2.01 (m, 2H), 1.38ESI(m/z):311.8[M+H+]。
Embodiment 13
In the reaction bulb with stirring and thermometer,31g (0.1mol) is added to methanolIn 500mL and 1 12mol/L HCl/1,4- dioxane 150mL, room temperature reaction is stayed overnight, and is spin-dried for, ether washing, is obtained22g
Embodiment 14
, will in the reaction bulb with stirring and thermometer21g (0.1mol) is added to Non-aqueous processingTHF 400mL in, add the THF200mL and m-trifluoromethyl phenyl boric acid 20g dissolved with potassium phosphate 60g (0.3mol)(0.12mol), is heated to 100 DEG C, reaction is stayed overnight, and ethyl acetate 200mL extractive reaction liquid is used after TLC monitoring raw material reactions completelyThree times, organic phase drying is separated, is obtained after being spin-dried for through column chromatography for separation23g；1H NMR(400MHz,CDCl3)δ:7.50-7.48 (m, 2H), 7.19-7.17 (m, 2H), 7.07 (s, 1H), 6.43 (s, 1H), 4.01 (d, J=8.0Hz,2H),3.33(s,2H),2.95(s,2H),2.02-2.01(m,2H).MS-ESI(m/z):321.3[M+H+]。
Embodiment 15
In reaction bulb,32g (0.1mol) is added in DMF, adds triethylamine 20g(0.2mol) and benzoic acid 12g (0.1mol), is heated to after 70 DEG C, reaction 3h through the reaction of TLC monitoring raw materials completely, reaction solutionIt is poured into water, with chloroform 200mL extractive reactions liquid three times, is spin-dried for obtaining compound after merging organic phase35g；1H NMR(400MHz,CDCl3)δ:8.03(s,1H),7.63-7.61(m,3H),7.50-7.48 (m, 2H), 7.19-7.17 (m, 2H), 7.07 (s, 1H), 6.43 (s, 1H), 4.09 (d, J=8.0Hz, 2H), 3.36 (s,2H),2.97(s,2H),2.07-2.06(m,2H).MS-ESI(m/z):425.4[M+H+]。
Embodiment 14
Antitumor activity is tested
Growth period breast cancer cell MCF-7 and liver cancer HepG2 are collected, the active anticancer of compound is determined with MTS methods,By cell with (every milliliter 4 × 10 of debita spissitudo4Individual cell) it is added in 96 porocyte culture plates and (must trains containing 10% tire calf serumNutrient solution is made into individual cells suspension), after cultivating 24 hours, in 37 DEG C, the CO that volumetric concentration is 5%2Under the conditions of with various concentrationsCompound effects 72 hours, then by MTS (final mass concentration 2mg/mL) and DMS (final 30 μM of molar concentration) mixingThing is directly added into celliferous culture medium, continues to put incubator incubation 4h.Act on after 4h, abandoning supernatant, 150 are added per holeμ LDMSO, vibration, suction of the metabolin that cell survival rate is acted on MTS by it under enzyme linked immunological monitor 490nm wavelengthYield is determined.
Preliminary biological activity test shows that such compound has suppression to make in cancer cell in breast cancer cell MCF-7With being less than MCF-7 to HepG2 inhibiting rate.
In summary, the invention provides a kind of new piperidines with antitumor activity and pyridine compounds and their and its systemPreparation Method, this is the discovery first of such compound purposes, with great research and development potentiality.
Embodiment above describes general principle, principal character and the advantage of the present invention, the technical staff of the industry shouldUnderstand, the present invention is not limited to the above embodiments, the original for simply illustrating the present invention described in above-described embodiment and specificationReason, under the scope for not departing from the principle of the invention, various changes and modifications of the present invention are possible, and these changes and improvements are each fallen withinIn the scope of protection of the invention.