CN104829554A - Phenothiazine compound and preparation method and application thereof - Google Patents
Phenothiazine compound and preparation method and application thereof Download PDFInfo
- Publication number
- CN104829554A CN104829554A CN201510271507.5A CN201510271507A CN104829554A CN 104829554 A CN104829554 A CN 104829554A CN 201510271507 A CN201510271507 A CN 201510271507A CN 104829554 A CN104829554 A CN 104829554A
- Authority
- CN
- China
- Prior art keywords
- add
- preparation
- group
- phenothiazine compound
- dried
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- XJEPALMCQLTXJA-QPJJXVBHSA-N C/C=C/Nc1c(C)ccc(C)c1 Chemical compound C/C=C/Nc1c(C)ccc(C)c1 XJEPALMCQLTXJA-QPJJXVBHSA-N 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D279/00—Heterocyclic compounds containing six-membered rings having one nitrogen atom and one sulfur atom as the only ring hetero atoms
- C07D279/10—1,4-Thiazines; Hydrogenated 1,4-thiazines
- C07D279/14—1,4-Thiazines; Hydrogenated 1,4-thiazines condensed with carbocyclic rings or ring systems
- C07D279/18—[b, e]-condensed with two six-membered rings
- C07D279/22—[b, e]-condensed with two six-membered rings with carbon atoms directly attached to the ring nitrogen atom
- C07D279/24—[b, e]-condensed with two six-membered rings with carbon atoms directly attached to the ring nitrogen atom with hydrocarbon radicals, substituted by amino radicals, attached to the ring nitrogen atom
- C07D279/28—[b, e]-condensed with two six-membered rings with carbon atoms directly attached to the ring nitrogen atom with hydrocarbon radicals, substituted by amino radicals, attached to the ring nitrogen atom with other substituents attached to the ring system
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D279/00—Heterocyclic compounds containing six-membered rings having one nitrogen atom and one sulfur atom as the only ring hetero atoms
- C07D279/10—1,4-Thiazines; Hydrogenated 1,4-thiazines
- C07D279/14—1,4-Thiazines; Hydrogenated 1,4-thiazines condensed with carbocyclic rings or ring systems
- C07D279/18—[b, e]-condensed with two six-membered rings
- C07D279/22—[b, e]-condensed with two six-membered rings with carbon atoms directly attached to the ring nitrogen atom
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D417/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00
- C07D417/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing two hetero rings
- C07D417/06—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing two hetero rings linked by a carbon chain containing only aliphatic carbon atoms
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Plural Heterocyclic Compounds (AREA)
Abstract
The invention relates to a phenothiazine compound and a preparation method thereof. The invention further relates the application of the phenothiazine compound in anti-cancer drug preparation and anti-cancer drugs using the compound as the effective component. The phenothiazine compound has the advantages that the compound with broad-spectrum anti-cancer effect is obtained by modifying phenothiazine ternary interlink parent nucleuses, synthesizing method is simple, and high yield is achieved; the phenothiazine compound has certain restraining effect on human breast cancer cells line MCF-7 and human hepatoma cell line Hep-G2, and a new thought is provided to new drugs satisfying clinic requirements.
Description
Technical field
The invention belongs to cancer therapy drug field, relate to a kind of the have phenothiazine compound of antitumous effect and the preparation method of this compound, in addition, the invention still further relates to this compounds and preparing the application in cancer therapy drug, and containing the cancer therapy drug of this compounds as effective constituent.
Background technology
Past, over more than 20 year, the cancer stem cell of various solid tumor and leukemia was also separated successively.Cancer stem cell (CSC:CancerStemCell) refers to the sub-fraction cell subsets (about 0.1%-5%) in tumour cell, there is the feature being similar to stem cell, have the features such as self, infinite multiplication and high tumorigenicity, great majority are in dormant state (G
0), have extremely strong resistance, traditional chemotherapy or radiotherapy can not be killed it, finally cause tumor recurrence and transfer.Carry out killing the experiment made on the living of breast carcinoma stem cell with Salinomycin. from the hoary hair institute of Harvard University in 2009 and Massachusetts Institute of Technology, the research of small molecule anti-cancer disease stem cell drugs obtains develop rapidly.Up to now, existing nearly 30 anticancer stem cell drugs are pushed to clinical one, two, three phases.
Phenothiazines be widely used, comparatively ripe a kind of antipsychotic class medicine, it mainly acts on the Dopamine Receptors of maincenter, has calmness, town is told, the effect such as antipsychotic and reduction body temperature.Research in recent years shows, phenothiazines has the effect of Tumor suppression stem cell.US Patent No. 2013/0065887A1 discloses a kind of dopamine-receptor antagonist being used for the treatment of acute myelogenous leukemia, this antagonist is phenothiazines thioridazine or chlorine pyridazine, it has selection targeting, can damage leukemia stem cell targetedly and not encroach on other cells.In the research of phenothiazines process lymphoma cell, have report display, under clinical common dose, chlorpromazine and trifluoperazine can promote lymphoma cell apoptosis and on normal lymphocytes without impact.Chinese patent application CN104114175 discloses the medical composition for removing cancer stem cell, and said composition comprises phenothiazines such as trifluoperazine can suppress lung cancer stem cell.Research in the past also finds that the trouble cancer probability calibration ordinary person of schizophreniac is low, also confirms that this compounds has potential anti-tumour effect.Above evidence all shows, phenothiazine compound and derivative thereof have the effect of Tumor suppression propagation or Tumor suppression stem cell.Thus one of direction that industry worker makes great efforts specially to grind has been become for the research of its broad spectrum anticancer effect of phenothiazine compound, have wide spectrum, novel phenothiazines that is efficient, low toxicity is study hotspot place, designing and screening this kind anti-cancer drugs with new chemical structure, novel mechanism or new role target position becomes problem demanding prompt solution.
Summary of the invention
The present inventor's process is to the research of phenothiazine compound structure activity relationship and screening, synthesize the phenothiazine compound with antitumous effect, experiment proves, this compound all has certain restraining effect for MCF-7 cell strainHJ2mm and Human hepatoma cell line Hep-G2.
Technical scheme of the present invention is achieved in that a kind of phenothiazine compound, and this compound structure is as shown in logical formula I:
Wherein, A is piperazinyl, piperidyl, triazol radical, tetrazole base, pyrryl, guanidine radicals, morpholinyl and derivatives group thereof; R is C
2-3straight chain saturated alkyl; B can be independently Cl, methylthio group or trifluoromethyl.
Further, when B is methylthio group, A is piperazinyl, triazol radical, tetrazole base, pyrryl, guanidine radicals, morpholinyl and derivatives group thereof, or piperidyl or N (CH
3)
2; When B is trifluoromethyl, A is morpholinyl, piperidyl, pyrryl and derivatives group thereof, or piperazinyl; When B is Cl, A is piperidyl, triazol radical, tetrazole base, pyrryl, guanidine radicals, morpholinyl and derivatives group thereof, or piperazinyl.
The present invention also asks the preparation method protecting above-mentioned phenothiazine compound, and presoma has the chemical structure of logical formula II, and the T1 group of this presoma and ligand groups in organic solvent substitution reaction occur; Wherein T
1be selected from halogenopropane and derivatives group thereof, T
2be selected from Cl, methylthio group or trifluoromethyl;
Described ligand groups comprises: piperidyl, piperazinyl, triazol radical, tetrazole base, pyrryl, guanidine radicals, morpholinyl and derivatives group thereof.
The method of the above-mentioned phenothiazine compound of another kind of preparation is: M in organic solvent substitution reaction occurs with the compound with logical formula III;
Wherein, M be selected from the halothane that replaces by piperidyl or derivatives thereof group and derivative thereof, B can be independently Cl, methylthio group or trifluoromethyl.
The piperidyl of aforesaid method formula of (I) or (II), piperazinyl and derivatives group thereof are connected by amidation or esterification by one or continuous two amino acid.
Further, described organic solvent as the mixture of a kind of or above-mentioned solvent in ether, alkane, aromatic hydrocarbon, amides, nitrile, amine, alcohols or ketones solvent, as THF, toluene, DMF, acetonitrile, triethylamine, acetone or alcohol.
The invention provides the novelty teabag of above-mentioned phenothiazine compound.
On the one hand, described phenothiazine compound is for the preparation of the medicine of anti-human breast cancer cell line mcf-7.
On the other hand, described phenothiazine compound is for the preparation of the medicine of anti-human liver cancer cell strain Hep-G2.
A kind of anticancer pharmaceutical composition, wherein containing the arbitrary compound of above-mentioned phenothiazines or their mixture as effective constituent, and one or more pharmaceutically acceptable carriers.
Further, this pharmaceutical composition can make the pharmaceutically acceptable formulation of any one, injection, sprays, inhalation or oral preparation.
Part particular compound of the present invention is as shown in table 1.
The list of table 1 compound
The above-mentioned phenothiazine compound of the present invention's synthesis, synthetic method is simple, productive rate is high, by to a chain of parent nucleus 10 of thiodiphenylamine ternary, 2 modify, obtain having and desired broad spectrum anticancer effect, such phenothiazine compounds all has certain restraining effect for MCF-7 cell strainHJ2mm and Human hepatoma cell line Hep-G2, for the new drug meeting clinical demand provides new approaches.
Embodiment
Below by specific embodiment, the present invention will be described, but the present invention is not limited thereto.Experimental technique described in following embodiment, if no special instructions, is ordinary method; Described reagent and biomaterial, if no special instructions, all can obtain from commercial channels.
Embodiment 1: preparation 2-21-2
2-10：
Get 2-trifluoromethyl thiodiphenylamine (2.00g, 7.4831mmol) and be placed in 500mL flask, add through anhydrous K
2cO
3dry THF (100mL), then at N
2add NaH mineral oil (1.1973g, 29.9323mmol) and the bromo-3-chloropropane (2.9601mL, 29.9323mmol) of 1-of 60% under protection successively, spend the night in the backflow of 65 DEG C of bath temperature.Be cooled to room temperature, then reaction solution poured in mixture of ice and water, be extracted with ethyl acetate (300mL+100mL × 2), merge organic phase, use anhydrous MgSO
4dry, solids removed by filtration, then filtrate is spin-dried for, then adds ethyl acetate (50mL) dissolving, in solution, add silica-gel powder (20g) be spin-dried for, dry method loading crosses silica column purification, with ethyl acetate and sherwood oil gradient elution, collect product, be spin-dried for, vacuum-drying obtains product 2-10 (4.9444g, productive rate 69.8%).+c ESI Q1MS：[M+H
+]344.00。
2-21-2：
Get 2-10 (250mg, 0.7272mmol) and be placed in 30mL straight tube bottle, then add morpholine (95.0mg, 1.0908mmol), triethylamine (0.30mL, 2.1816mmol) and acetonitrile (10mL), stir and spend the night at 80 DEG C.Be cooled to room temperature, directly be spin-dried for, by thick product dissolve with methanol, filter, in filtrate, add silica-gel powder make solid solution, dry method loading crosses silica column purification, with triethylamine, ethyl acetate and sherwood oil gradient elution, collect product, be spin-dried for, vacuum-drying obtains product 2-21-2 (108.5mg, productive rate 31.5%).
1H-NMR(400MHz,CDCl
3)δ7.23-7.10(m,4H),7.04(s,1H),7.00-6.89(m,2H),4.00(t,J＝6Hz,2H),3.76-3.62(m,4H),2.61-2.39(m,6H),2.08-1.92(m,2H)；+c ESI Q1MS：[M+H
+]395.09。
Embodiment 2: preparation 2-21-6
In 30mL straight tube bottle, add 2-10 (250mg, 0.7272mmol), dried meat ammonia alcohol (110.3mg, 1.0908mmol), triethylamine (0.5mL) and acetonitrile (10mL), stir at 80 DEG C and spend the night.Be cooled to room temperature, directly be spin-dried for, by thick product dissolve with methanol, filter, in filtrate, add silica-gel powder make solid solution, dry method loading crosses silica column purification, with triethylamine, ethyl acetate and sherwood oil gradient elution, collect product, be spin-dried for, vacuum-drying obtains product 2-21-6 (138.9mg, productive rate 38.9%).
1H-NMR(400MHz,CDCl
3)δ7.25-7.13(m,4H),7.05(s,1H),7.01-6.89(m,2H),4.05-3.96(m,2H),3.68-3.60(m,1H),3.51-3.42(m,1H),3.33-3.23(m,1H),3.14-3.03(m,1H),2.81-2.69(m,1H),2.61-2.50(m,1H),2.35-2.24(m,1H),2.16-2.05(m,2H),1.85-1.72(m,3H),1.28-1.23(m,2H)；+c ESI Q1MS：[M+H
+]409.11。
Embodiment 3: preparation 2-163,2-172
2-21-1：
Get 2-10 (300mg, 0.8726mmol) be placed in 30mL straight tube bottle, then 1-tert-butoxycarbonyl-piperazine (243.8mg is added, 1.3089mmol), triethylamine (0.37ml, 2.6178mmol) with acetonitrile (10mL), react under 80 DEG C of conditions and spend the night.Be cooled to room temperature, direct evaporate to dryness, by thick product dissolve with methanol, then in filtrate, add silica-gel powder and make solid solution, dry method loading crosses silica column purification, with ethyl acetate, sherwood oil and triethylamine gradient wash-out, collect product, be spin-dried for, vacuum-drying obtains product 2-21-1 (63.4mg, productive rate 40.2%).
1H-NMR(400MHz,CDCl
3)δ7.24-7.10(m,5H),7.04(s,1H),7.00-6.88(m,1H),4.08-3.95(m,2H),3.60-3.25(m,4H),2.70-2.25(m,6H),2.12-1.90(m,2H),1.44(s,9H)；+c ESI Q1MS：[M+H
+]494.20。
2-163：
In 100mL flask, 2-21-1 (2.4458g, 4.9569mmol) is dissolved in CH
2cl
2(50mL), TFA (4mL) is added, room temperature for overnight.Concentrated except desolventizing and most of TFA, thick product dissolve with methanol, then adds NaHCO
3neutralize excessive TFA, solids removed by filtration, add silica-gel powder and make solid solution in filtrate, dry method loading crosses silica column purification, uses NH
3h
2o, CH
3oH and CH
2cl
2gradient elution, collect product, be spin-dried for, vacuum-drying obtains product 2-163 (2.4346g, productive rate 100%).
1H-NMR(400MHz,CDCl
3)δ＝7.23-7.10(m,4H),7.03(s,1H),6.99-6.93(t,J＝6Hz,1H),6.92-6.87(d,J＝8Hz,1H),3.99(t,J＝4Hz,2H),3.10-2.95(m,4H),2.70-2.47(m,6H),1.95-1.86(m,2H)。+c ESI Q1MS：[M+H
+]394.25。
2-169：
By 2-163 (1.9146g, 4.8681mmol) be placed in 200mL flask, then Boc-glycine (Boc-Gly) (0.9381g, 5.3549mmol) is added, HBTU (2.7404g, 7.3022mmol), HOBT (986.7mg, 7.3022mmol) and DMF (50mL), drips DIEA (3.8159mL under 0 DEG C of condition, 21.9065mmol), then move to room temperature for overnight.Reaction solution is poured into saturated NaHCO
3in solution (500mL), with ethyl acetate (500mL) extraction, phase-splitting that aqueous phase is extracted with ethyl acetate again (200mL × 3); Merge organic phase, then use saturated NaHCO
3solution cleaning (200mL × 2), then clean (200mL × 2) by saturated NaCl solution; The anhydrous MgSO of organic phase after phase-splitting
4drying, solids removed by filtration, adds silica-gel powder and makes solid solution in filtrate, dry method loading crosses silica column purification, with ethyl acetate and sherwood oil gradient elution, collects product, be spin-dried for, vacuum-drying obtains product 2-169 (2.6138g, productive rate 97.54%).
1H-NMR(400MHz,CDCl
3)δ7.21-7.10(m,4H),7.03(s,1H),6.97-6.89(m,2H),3.99(t,J＝4Hz,2H),3.93-3.88(d,J＝4Hz,2H),3.56-3.51(m,2H),3.29-3.24(m,2H),2.48(t,J＝6Hz,2H),2.40-2.33(m,4H),1.95-1.88(m,2H),1.44(s,9H)
2-172：
2-169 (2.5949g, 4.7140mmol) is placed in 100mL flask, then adds CH
2cl
2(20mL) dissolve, then add TFA (5mL) stir spend the night.Be spin-dried for, by thick product CH
3oH dissolves, and then adds NaHCO
3neutralize unnecessary TFA; Solids removed by filtration, is spin-dried for filtrate, and thick product uses CH again
2cl
2dissolve, wet method loading crosses silica column purification, uses NH
3h
2o, methyl alcohol and methylene dichloride carry out gradient elution, and collect product, be spin-dried for, vacuum-drying obtains product 2-172 (1.3303g, productive rate 62.8%).
1H-NMR(400MHz,CDCl
3)δ7.22-7.11(m,4H),7.04(s,1H),6.98-6.89(m,2H),4.00(t,J＝6Hz,2H),3.56(t,J＝4Hz,2H),3.42(s,2H),3.27(t,J＝4Hz,2H),2.49(t,J＝4Hz,2H),2.37(t,J＝4Hz,4H),1.97-1.89(m,2H),1.25(s,2H)；+c ESI Q1MS：[M+H
+]451.39。
Embodiment 4: preparation 2-21-4,2-162
2-21-4：
With 2-10 (250mg, 0.7272mmol), piperidines-4-alcohol (110.3mg, 1.0908mmol), triethylamine (0.5mL) and acetonitrile (10mL), stir at 80 DEG C and spend the night.Be cooled to room temperature, directly be spin-dried for, by thick product dissolve with methanol, filter, in filtrate, add silica-gel powder make solid solution, dry method loading crosses silica column purification, with triethylamine, ethyl acetate and sherwood oil gradient elution, collect product, be spin-dried for, vacuum-drying obtains product 2-21-4 (232.5mg, productive rate 65.1%).
1H-NMR(400MHz,CDCl
3)δ7.24-7.10(m,4H),7.04(s,1H),6.99-6.90(m,2H),3.97(t,J＝6Hz,2H),3.79-3.66(m,2H),2.82-2.72m,2H),2.60-2.50(m,2H),2.32-2.14(m,2H),1.98-1.86(m,2H),1.63-1.50(m,2H),1.14(t,J＝8Hz,2H)；+c ESI Q1MS：[M+H
+]409.13。
2-154：
Get 2-21-4 (1.6316g, 3.9947mmol) and be placed in 250mL flask, add Boc-Gly (839.7mg, 4.7936mmol), DMAP (48.8mg, 0.3995mmol) and CH
2cl
2(200mL), stir 30 minutes at reaction solution being placed in 0 DEG C, then add DCC (2.4727g, 11.9841mmol), then go to room temperature for overnight.Solids removed by filtration impurity; Add methyl alcohol to be dissolved by thick product, add silica-gel powder and make solid solution in solution, dry method loading crosses silica column purification, with ethyl acetate and sherwood oil gradient elution, collect product, be spin-dried for, vacuum-drying obtains product 2-154 (3.2987g, productive rate 100%).
1H-NMR(400MHz,CDCl
3)δ7.21-7.09(m,4H),7.04(s,1H),6.97-6.89(m,2H),4.86-4.77(m,1H),3.96(t,J＝4Hz,2H),2.70-2.58(m,2H),2.52-2.42(m,2H),2.28-2.16(m,2H),1.98-1.89(m,4H),1.87-1.80(m,2H),1.77-1.70(m,4H),1.45(s,9H)；+c ESI Q1MS：[M+H
+]551.42。
2-162：
Get 2-154 (3.1691g, 5.6029mmol) and be placed in 100mL flask, add CH
2cl
2(50mL) dissolve, then add TFA (2.15mL, 28.0143mmol), room temperature for overnight.Be spin-dried for removing CH
2cl
2, by thick product dissolve with methanol, under stirring, add NaHCO
3neutralize unnecessary TFA, solids removed by filtration, add silica-gel powder and make solid solution in filtrate, dry method loading crosses silica column purification, uses NH
3h
2o, CH
3oH and CH
2cl
2carry out gradient elution, collect product, be spin-dried for, vacuum-drying obtains product 2-162 (1.5027g, productive rate 57.7%).
1H-NMR(400MHz,CDCl
3)δ7.22-7.10(m,4H),7.04(s,1H),6.98-6.90(m,2H),4.84-4.75(m,1H),3.96(t,J＝4Hz,2H),3.41(s,2H),2.70-2.60(m,2H),2.47(t,J＝6Hz,2H),2.25-2.16(m,2H),1.97-1.89(m,2H),1.89-1.81(m,2H),1.68-1.57(m,4H)；+c ESI Q1MS：[M+H
+]466.38。
Embodiment 5: preparation 4-215
7-78：
2-(piperidin-2-yl) ethane-1-alcohol (20.00g, 154.8947mmol) is placed in 500mL flask, adds CH
2cl
2(300mL) dissolve, then add triethylamine (45mL, 309.7894mmol), under stirring, add Boc
2o (40.5669g, 185.8736mmol), reacts 4 hours under room temperature.Directly be spin-dried for, by thick product CH
3oH dissolves, and adds NaHCO
3neutralize unnecessary TFA, filter, in filtrate, add silica-gel powder make solid solution, dry method loading crosses silica column purification, with ethyl acetate and sherwood oil gradient elution, collects product, concentrated, vacuum-drying obtains product 7-78 (36.2772g, productive rate 100%).ITMS+c ESI[M+H
+]230.18。
7-80：
Get 7-78 (15.00g, 65.4094mmol) and be placed in 500mL flask, add through anhydrous K
2cO
3dry THF (150mL), triphenyl phosphorus (27.7142g, 105.6623mmol), add NBS (18.8058g, 105.6623mmol) under stirring in batches, be transferred to stirred at ambient temperature 3 days at 0 DEG C.Directly be spin-dried for, thick product CH
2cl
2dissolve, in solution, add silica-gel powder make solid solution, dry method loading crosses silica column purification, and with ethyl acetate and sherwood oil gradient elution, collect product, concentrated, vacuum-drying obtains product 7-80 (12.0956g, productive rate 63.26%).
1H-NMR(500MHz,CDCl
3)δ4.44-4.35(m,1H),4.09-3.94(m,1H),3.41-3.28(m,2H),2.80-2.66(m,1H),2.40-2.29(m,1H),1.98-1.84(m,1H),1.76-1.61(m,3H),1.58-1.43(m,12H)。
4-197：
Get trifluoperazine (1.00g, 3.7415mmol) and be placed in 100mL tri-neck round-bottomed flask, add through the dried THF (50mL) of Anhydrous potassium carbonate, at N
2add NaH (359.2mg, 14.966mmol), the 7-80 (1.0975g, 3.7415mmol) of the mineral oil protection of 60% under protection, flow through night next time in 60 DEG C of heating conditions.After reaction terminates, stopped reaction, be cooled to room temperature, reaction solution is poured in 200mL frozen water, be extracted with ethyl acetate (200mL × 3), merge organic phase anhydrous magnesium sulfate drying, filter, in filtrate, add (3g) silica-gel powder make solid solution, dry method loading crosses silica column purification, silicagel column: 2.5 × 20cm, does solvent homogenate wet method dress post with sherwood oil, dry method loading, with ethyl acetate and sherwood oil gradient elution, collect product, be spin-dried for, vacuum-drying obtains sterling 4-197 (1.40g, productive rate 78%).ITMS+c ESI[M+H
+]479.12。4-215：
Get 4-197 (1.30g, 2.7050mmol) and be placed in 100mL flask, add methylene dichloride (50mL), add TFA (2.0727ml, 27.0498mmol) under stirring, stirred overnight at room temperature.Direct evaporate to dryness, by thick product dissolve with methanol, adds NaHCO
3neutralize unnecessary TFA; Filter, in filtrate, add silica-gel powder make solid solution, cross silica column purification.Use NH
3h
2o, methyl alcohol and dichloromethane gradient drip washing, collect product, be spin-dried for, vacuum-drying obtains product 4-215 (539.2mg, productive rate 53%).
1H-NMR(500MHz,CDCl
3)δ7.26-7.14(m,4H),7.10-7.07(m,1H),7.01-6.92(m,2H),4.08-3.94(m,3H),3.12-3.02(m,1H),2.73-2.59(m,2H),1.92-1.85(m,2H),1.84-1.76(m,1H),1.72-1.64(m,2H),1.44-1.34(m,2H)；ITMS+c ESI：[M+H
+]379.20。
Embodiment 6 prepares 2-30-3
2-133：
Get 2-methylthio group thiodiphenylamine (10.00g, 40.8163mmol) and be placed in 500mL flask, add through anhydrous K
2cO
3dry THF (250mL), at N
2add NaH (6.5306g, 163.2653mmol) and the bromo-3-chloropropane (16.15mL, 163.2653mmol) of 1-of the mineral oil protection of 60% under protection, flow through night next time at 65 DEG C.Be cooled to room temperature, poured into by reaction solution in frozen water, thick product is extracted with ethyl acetate (300mL+100mL × 2), merges the anhydrous MgSO of organic phase
4drying, solids removed by filtration, is spin-dried for, and then adds ethyl acetate (50mL) and is dissolved by set product, add silica-gel powder (20g) and make solid solution in solution.Dry method loading crosses silica column purification, and with ethyl acetate and sherwood oil gradient elution, collect product, be spin-dried for, vacuum-drying obtains product 2-133 (13.798g, productive rate 100%).
1H-NMR(400MHz,CDCl
3)δ7.20-7.14(m,2H),7.07(d,J＝8Hz,1H),6.97-6.88(m,2H),6.87-6.81(m,2H),4.08(t,J＝6Hz,2H),3.67(t,J＝6Hz,2H),2.47(s,3H),2.27-2.21(m,2H)；+c ESI Q1MS[M+H
+]322.13。
2-30-3：
With 2-133 (300mg, 0.9320mmol), dimethylamine hydrochloride (114mg, 1.3980mmol), triethylamine (0.39mL, 2.7961mmol) and acetonitrile (10mL), stir at 80 DEG C and spend the night.Be cooled to room temperature, directly be spin-dried for, by thick product dissolve with methanol, filter, in filtrate, add silica-gel powder make solid solution, dry method loading crosses silica column purification, with triethylamine, ethyl acetate and sherwood oil gradient elution, collect product, be spin-dried for, vacuum-drying obtains product 2-30-3 (83.2mg, productive rate 24.3%).
1H-NMR(400MHz,CDCl
3)δ7.18-7.10(m,2H),7.07-7.01(m,1H),6.94-6.86(m,2H),6.84-6.78(m,3H),3.93-3.85(m,2H),2.46(s,3H),2.42-2.36(m,2H),2.22-2.19(s,6H),1.98-1.90(m,2H)；+c ESI Q1MS:[M+H
+]331.10,[M+NH
3]347.08。
Embodiment 7 prepares 2-30-2
Get 2-133 (300mg, 0.9320mmol), morpholine (121.8mg, 1.3981mmol), triethylamine (0.39mL, 2.7961mmol) and acetonitrile (10mL), stir at 80 DEG C and spend the night.Be cooled to room temperature, directly be spin-dried for, by thick product dissolve with methanol, filter, in filtrate, add silica-gel powder make solid solution, dry method loading crosses silica column purification, with triethylamine, ethyl acetate and sherwood oil gradient elution, collect product, be spin-dried for, vacuum-drying obtains product 2-30-2 (121mg, productive rate 33.5%).
1H-NMR(400MHz,CDCl
3)δ7.18-7.09(m,2H),7.08-7.01(m,1H),6.95-6.86(m,2H),6.85-6.78(m,2H),3.94(t,J＝6Hz,2H),3.71-3.61(m,4H),2.51-2.34(m,9H),1.99-188(m,2H)；+c ESI Q1MS:[M+H
+]373.23。
Embodiment 8 prepares 2-30-4
With 2-133 (300mg, 0.9320mmol), piperidines-4-alcohol (141.4mg, 1.3979mmol), triethylamine (0.39mL, 2.7961mmol) and acetonitrile (10mL), stir at 80 DEG C and spend the night.Be cooled to room temperature, directly be spin-dried for, by thick product dissolve with methanol, filter, in filtrate, add silica-gel powder make solid solution, dry method loading crosses silica column purification, with triethylamine, ethyl acetate and sherwood oil gradient elution, collect product, be spin-dried for, vacuum-drying obtains product 2-30-4 (126.1mg, productive rate 35.0%).+c ESI Q1MS:[M+H
+]387.16。
Embodiment 9 prepares 2-30-5
With 2-133 (300mg, 0.9320mmol) be placed in 30ml straight tube bottle, add 1-(2-hydroxyethyl) pyrazine (182mg, 1.3980mmol), triethylamine (0.39mL, 2.7961mmol) with acetonitrile (10mL), stir at 80 DEG C and spend the night.Stopped reaction, be cooled to room temperature, be spin-dried for, by thick product dissolve with methanol, in filtrate, add silica-gel powder make solid solution, dry method loading crosses silica column purification, with triethylamine, ethyl acetate and sherwood oil gradient elution, collect product, be spin-dried for, vacuum-drying obtains product 2-30-5 (106mg, productive rate 27.4%).+c ESI Q1MS:[M+H
+]416.15。
Embodiment 10 prepares 2-30-6
Get 2-133 (300mg, 0.9320mmol) and be placed in 30mL straight tube bottle, add dried meat ammonia alcohol (141.4mg, 1.3979mmol), triethylamine (0.39mL, 2.7961mmol) and acetonitrile (10mL), stir and spend the night at 80 DEG C.Be cooled to room temperature, be spin-dried for, by thick product dissolve with methanol, in filtrate, add silica-gel powder make solid solution, dry method loading crosses silica column purification, with triethylamine, ethyl acetate and sherwood oil gradient elution, collect product, be spin-dried for, vacuum-drying obtains product 2-30-6 (88.3mg, productive rate 24.5%).+c ESI Q1MS:[M+H
+]387.16。
Embodiment 11 prepares 2-58
Get 2-133 (0.3126g, 0.9712mmol), be placed in 30mL straight tube bottle, add alcohol solvent (20mL), Guanidinium hydrochloride (185.5mg, 1.9423mmol), then NaOH (77.7mg, 1.9423mmol) is dissolved in H
2add again in O (1mL) in straight tube bottle, stir under 80 DEG C of conditions after sealing and spend the night.Wet method loading crosses silica column purification, and with ammoniacal liquor, methyl alcohol and dichloromethane gradient drip washing, collect product, be spin-dried for, vacuum-drying obtains product 2-58 (141mg, productive rate 38.1%).+c ESI Q1MS:[M+H
+]345.30。
Embodiment 12 prepares 2-60
2-133 (0.2964g, 0.9208mmol) is joined in 30mL straight tube bottle, add 1,2,4-triazole (0.1272g, 1.8417mmol), K
2cO
3(0.7636g, 5.5250mmol), KI (15.3g, 0.09208mmol) and acetone (10mL), stir after sealing and spend the night under 60 DEG C of conditions.Filter, filtrate is spin-dried for, thick product methylene dichloride is dissolved, in solution, adds silica-gel powder make solid solution, dry method loading crosses silica column purification, with methyl alcohol and dichloromethane gradient drip washing, collects product, be spin-dried for, vacuum-drying obtains product 2-60 (179.0mg, productive rate 54.8%).
1H-NMR(400MHz,CDCl
3)δ7.94(s,1H),7.88(s,1H),7.24-7.10(m,3H),7.04-6.94(m,1H),6.90-6.78(m,2H),6.77-6.74(m,1H),4.35-4.24(m,2H),3.85-3.74(m,2H),2.50-2.34(m,5H)；+c ESIQ1MS:[M+H
+]355.18，[M+Na
+]377.11。
Embodiment 13 prepares 2-138
2-133 (300mg, 0.9320mmol) is joined in 100ml flask, adds 5-methyl isophthalic acid H-tetrazole (156.6mg, 1.8640mmol), K
2cO
3(0.7729g, 5.5921mmol), KI (15.4mg, 0.09320mmol) and acetone (15ml).Under 60 DEG C of conditions, stirring and refluxing is spent the night.Be cooled to room temperature, filter, in filtrate, add silica-gel powder make solid solution, dry method loading crosses silica column purification, and with ethyl acetate and sherwood oil gradient elution, collect product, be spin-dried for, vacuum-drying obtains product 2-138 (71.3mg, productive rate 20.7%).
1H-NMR(400MHz,CDCl
3)δ7.20-7.11(m,2H),7.09(d,J＝8Hz,1H),6.97-6.92(m,1H),6.89-6.80(m,2H),6,76-6.74(m,1H),4.68(t,J＝6Hz,2H),3.98(t,J＝6Hz,2H),2.52-2.44(m,8H)。
Embodiment 14 prepares 2-139
2-133 (0.2964g, 0.9208mmol) is joined in 100mL flask, adds 1,2,3-triazole (0.11mL, 1.8640mmol), K
2cO
3(0.7729g, 5.5961mmol), KI (15.4mg, 0.09320mmol) and acetone (15mL).Under 60 DEG C of conditions, stirring and refluxing is spent the night.Be cooled to room temperature, filter, add silica-gel powder after filtrate is concentrated and make solid solution, dry method loading crosses silica column purification, with ethyl acetate and sherwood oil gradient elution, collects product, be spin-dried for, vacuum-drying obtains product 2-139 (202.5mg, productive rate 61.3%).
1H-NMR(400MHz,CDCl
3)δ7.59-7.58(m,2H),7.21-7.02(m,3H),7.00-6.68(m,4H),4.59(t,J＝6Hz,2H),4.02-3.84(m,2H),2.54-2.36(m,5H)。
Embodiment 15 prepares 2-149
2-133 (300mg, 0.9320mmol) is joined in 100mL flask, adds 1H-tetrazole (0.1306mg, 1.8640mmol), K
2cO
3(0.7729g, 5.5921mmol), KI (15.4mg, 0.09320mmol) and acetone (50mL).Stir under 60 DEG C of conditions and spend the night.Be cooled to room temperature, filter, in filtrate, add silica-gel powder make solid solution, dry method loading crosses silica column purification.With ethyl acetate and sherwood oil gradient elution, collect product, be spin-dried for, vacuum-drying obtains product 2-149 (44.1mg, productive rate 13.3%).+c ESI Q1MS:[M+H
+]356.13,[M+Na
+]378.11。
Embodiment 16 prepares 2-72
2-30-1：
Get 2-12 (300mg, 0.9320mmol) be placed in 30mL straight tube bottle, then 1-tert-butoxycarbonyl-piperazine (260.4mg is added, 1.3980mmol), triethylamine (0.39mL, 2.7961mmol) with acetonitrile (10mL), stir under 80 DEG C of conditions after sealing and spend the night.Be cooled to room temperature, be spin-dried for, by thick product dissolve with methanol, in filtrate, add silica-gel powder make solid solution, dry method loading crosses silica column purification, with triethylamine, ethyl acetate and sherwood oil gradient elution, collect product, be spin-dried for, vacuum-drying obtains product 2-30-1 (75.1mg, productive rate 17.08%).+c ESI Q1MS:[M+H
+]472.13,[M+NH
3]488.11。
2-72：
Get 2-30-1 (75.1mg, 0.1522mmol) and be placed in 100mL flask, add CH
2cl
2(50mL) dissolve, then add TFA (4mL, 52.2014mmol), room temperature for overnight.Be spin-dried for, by thick product dissolve with methanol, then add NaHCO
3neutralize unnecessary TFA, filter, in filtrate, add silica-gel powder make solid solution, dry method loading crosses silica column purification, uses NH
3h
2o, CH
3oH and CH
2cl
2gradient elution, collect product, be spin-dried for, vacuum-drying obtains product 2-72 (46.2mg, productive rate 84.5%).
1H-NMR(400MHz,CDCl
3)δ7.18-7.10(m,2H),7.05(d,J＝8Hz,1H),6.95-6.76(m,4H),3.94(t,J＝6Hz,2H),3.10-3.02(m,4H),2.71-2.59(m,4H),2.58-2.50(m,2H),3.46(s,3H)；+c ESI Q1MS[M+H
+]373.23。
Embodiment 17 prepares 7-4
7-2：
Get Boc-Gly (0.8453g, 4.8251mmol) and be placed in 500mL flask, add 2-30-4 (1.6955g, 4.3865mmol), HBTU (2.4693g, 6.5798mmol) and HOBT (0.8891g, 6.5798mmol); Under 0 DEG C of condition, drip DIEA (3.4384mL, 19.7393mmol), after 30 minutes, go to room temperature for overnight.Directly pour reaction solution into saturated NaHCO
3in solution (500mL), be extracted with ethyl acetate (500mL+200mL × 3+100mL), merge organic phase, first use saturated NaHCO
3solution cleaning (300mL × 2), then clean (300mL × 2) with saturated aqueous common salt; The anhydrous MgSO of organic phase
4drying, filter, in filtrate, add silica-gel powder make solid solution, dry method loading crosses silica column purification, and with ethyl acetate and sherwood oil gradient elution, collect product, be spin-dried for, vacuum-drying obtains product 7-2 (2.3893g, productive rate 100%).
1H-NMR(400MHz,CDCl
3)δ7.20-7.10(m,2H),7.08-7.02(m,1H),6.96-6.86(m,2H),6.85-6.78(m,2H),4.88-4.81(m,1H),3.96-3.91(m,2H),2.75-2.65(m,2H),2.60-2.50(m,2H),2.47(s,3H),2.40-2.25(m,2H),2.02-1.84(m,4H),1.82-1.63(m,4H),1.45(s,9H)；+c ESI Q1MS:[M+H
+]544.32。
7-4：
7-2 (2.3082g, 4.2453mmol) is placed in 200mL flask, adds CH
2cl
2(15mL), TFA (3.25mL, 42.4527mmol) is added under magnetic agitation, room temperature for overnight.Be spin-dried for, by thick product CH
3oH dissolves, and adds NaHCO
3neutralize unnecessary TFA, filter, in filtrate, add silica-gel powder make solid solution, dry method loading crosses silica column purification, uses NH
3h
2o, CH
3oH and CH
2cl
2gradient elution, collect product, be spin-dried for, vacuum-drying obtains product 7-4 (896.6mg, productive rate 47.7%).
1H-NMR(400MHz,CDCl
3)δ7.18-7.10(m,2H),7.08-7.02(m,1H),6.94-6.87(m,2H),6.85-6.78(m,2H),4.85-4.77(m,1H),3.92(t,J＝4Hz,2H),3.41(s,2H),2.72-2.62(m,2H),2.52-2.43(m,5H),2.29-2.16(m,2H),2.00-1.83(m,4H),1.73-1.62(m,4H)。
Embodiment 18 prepares 7-86,7-92,7-147
7-82：
Get 2-methylthio group thiodiphenylamine (10.1029g, 41.2362mmol) and be placed in 500mL flask, add through anhydrous K
2cO
3dried THF (200mL), at N
2lower NaH (6.5978g, 164.9448mmol) and the 7-80 (12.0956g, 41.2362mmol) successively adding the mineral oil protection of 60% of protection.Night is flow through next time 60 DEG C of conditions.Be cooled to room temperature, reaction solution poured in mixture of ice and water, be extracted with ethyl acetate (500mL × 3), merge the anhydrous MgSO of organic phase
4drying, filter, in filtrate, add silica-gel powder make solid solution, dry method loading crosses silica column purification, with ethyl acetate and sherwood oil gradient elution, is spin-dried for, and vacuum-drying obtains product 7-82 (19.1176g, productive rate 100%).
1H-NMR(500MHz,CDCl
3)δ7.18-7.12(m,2H),7.08-7.03(m,1H),6.96-6.89(m,1H),6.88-6.80(m,2H),6.79-6.74(m,1H),4.41(s,1H),4.10-3.95(m,1H),3.94-3.75(m,2H),2.80(t,J＝15Hz,1H),2.46(s,3H),2.28-2.14(m,1H),1.94-1.82(m,1H),1.61-1.53(m,3H),1.51-1.34(m,12H)。7-86：
In 100mL flask, 7-82 (19.00g, 41.5459mmol) is dissolved in methylene dichloride (20mL), adds TFA (15.92mL, 207.7297mmol), at room temperature stir and spend the night.Be spin-dried for, by thick product dissolve with methanol, then add NaHCO
3neutralize unnecessary TFA, filter, in filtrate, add silica-gel powder make solid solution, cross silica column purification.Use NH
3h
2o, CH
3oH and CH
2cl
2gradient elution, collect product, be spin-dried for, vacuum-drying obtains product 7-86 (19.0062g, productive rate 100%).
1H-NMR(500MHz,CDCl
3)δ7.20-7.13(m,2H),7.11-7.06(m,1H),6.98-6.92(m,1H),6.88-6.82(m,2H),6.80-6.77(m,1H),3.96(t,J＝7.5Hz,2H),3.22-3.12(m,1H),3.00-2.92(m,1H),2.74-2.64(m,1H),2.46(s,3H),2.25-2.13(m,1H),2.02-1.90(m,1H),1.85-1.75(m,1H),1.72-1.55(m,3H),1.54-1.33(m,2H)。
7-88：
Get Boc-Gly (10.2309g, 58.4025mmol) be placed in 500mL flask, add 7-86 (18.9118g, 53.0932mmol), HBTU (29.8880g, 79.6398mmol), HOBT (10.9609g, 79.7398mmol) and DMF (200mL), drips DIEA (41.62mL under 0 DEG C of condition, 238.9194mmol), room temperature for overnight is gone to after 30 minutes.Directly pour reaction solution into saturated NaHCO
3in solution (500mL), be extracted with ethyl acetate (500mL+200mL × 3+100mL), merge the saturated NaHCO of organic phase
3solution cleaning (300mL × 2), then clean (300mL × 2) with the saturated common salt aqueous solution.The anhydrous MgSO of organic phase
4drying, filters, adds silica-gel powder and make solid solution in filtrate, crosses silica column purification.With ethyl acetate and sherwood oil gradient elution, collect product, be spin-dried for, vacuum-drying obtains product 7-88 (19.1364g, productive rate 70.2%).
1H-NMR(500MHz,CDCl
3)δ7.22-7.02(m,3H),6.97-6.89(m,1H),6.88-6.80(m,2H),6.79-6.70(m,1H),4.05-3.80(m,4H),3.75-3.65(m,1H),3.60-3.40(m,1H),3.14-3.05(m,1H),2.47(s,3H),2.25-2.10(m,1H),1.95-1.85(m,1H),1.62-1.30(m,15H)。
7-92：
7-88 (19.0825g, 37.1703mmol) is placed in 200mL flask, adds methylene dichloride (20mL) and dissolve, then add TFA (14.24mL, 185.8516mmol), room temperature for overnight.Be spin-dried for, by thick product dissolve with methanol, add NaHCO
3neutralize unnecessary TFA.Filter, in filtrate, add silica-gel powder make solid solution, cross silica column purification.Use NH
3h
2o, methyl alcohol and dichloromethane gradient drip washing, collect product, be spin-dried for, vacuum-drying obtains product 7-92 (13.6454g, productive rate 89.05%).
1H-NMR(500MHz,CDCl
3)δ7.24-7.10(m,2H),7.08-7.03(m,1H),7.00-6.77(m,3H),6.76-6.70(m,1H),3.93-3.81(m,2H),3.65-3.47(m,2H),3.40-3.32(m,1H),3.26-3.19(m,1H),3.13-3.04(m,1H),2.46(s,3H),2.25-2.05(m,2H),1.70-1.28(m,6H)。
7-145：
Get Boc-Gly (2.7978g, 15.9708mmol) be placed in 500ml flask, add 7-92 (6.00g, 14.5189mmol), HBTU (8.1732g, 21.7784mmol), HOBT (2.9427g, 21.7784mmol) and DMF (200mL); Under 0 DEG C of condition, drip DIEA (11.38mL, 65.3351mmol), after 30 minutes, go to room temperature for overnight.Directly pour reaction solution into saturated NaHCO
3in solution (500mL), be extracted with ethyl acetate (500mL+200mL × 3+100mL), merge the saturated NaHCO of organic phase
3solution cleaning (200mL × 2), then clean (200mL × 2) with saturated aqueous common salt; The anhydrous MgSO of organic phase
4drying, filters, adds silica-gel powder and make solid solution in filtrate, crosses silica column purification.With ethyl acetate and sherwood oil gradient elution, collect product, be spin-dried for, vacuum-drying obtains product 7-145 (7.67g, productive rate 93%).
1H-NMR(500MHz,CDCl
3)δ7.22-6.60(m,9H),5.03-4.93(m,1H),4.05-3.70(m,6H),2.45(s,3H),1.72-1.56(m,6H),1.48(s,9H)。
7-147：
7-145 (7.57g, 13.2706mmol) is placed in 500mL flask, adds methylene dichloride (20mL) and dissolve, then add TFA (10.17mL, 132.7057mmol), room temperature for overnight.Be spin-dried for, by thick product dissolve with methanol, then add NaHCO
3neutralize unnecessary TFA, filter, in filtrate, add silica-gel powder make solid solution, dry method loading crosses silica column purification, uses NH
3h
2o, methyl alcohol and dichloromethane gradient drip washing, collect product, be spin-dried for, vacuum-drying obtains product 7-147 (5.3975g, productive rate 86.48%).
1H-NMR(500MHz,CDCl
3)δ7.80-7.55(m,1H),7.22-7.09(m,2H),7.08-7.02(m,1H),6.97-6.89(m,1H),6.87-6.79(m,2H),6.79-6.69(m,1H),5.03-4.94(m,1H),4.08-4.01(m,1H),3.96-3.85(m,2H),3.84-3.65(m,1H),3.58-3.47(m,1H),3.40-3.29(m,2H),3.16-3.06(m,1H),2.47(s,3H),2.27-2.08(m,1H),1.95-1.82(m,1H),1.62-1.13(m,6H)；ITMS+c ESI[M+H
+]571.19,[M+Na
+]593.31。
Embodiment 19 prepares 6-78
6-71：
Under nitrogen protection; 2-chloro phenothiazine (5.00g is added in 500ml flask; 21.3mmol); dissolve with THF; then add the NaH (4.2786g, 106.966mmol) of the mineral oil protection of 60%, release a large amount of bubble; then the bromo-3-chloropropane (10.57mL, 106.966mmol) of 1-is added.Reflux in the oil bath of 65 DEG C and spend the night.Stopped reaction, pours into reaction solution in mixture of ice and water (500mL), is extracted with ethyl acetate (300mL × 3), merges organic phase anhydrous magnesium sulfate drying, filters.Column chromatography, with ethyl acetate and sherwood oil gradient elution, collect product point, concentrating under reduced pressure obtains product 6-71 (6.6377g, productive rate 59.11%).+c ESI Q1 MS：[M+H
+]310.10
6-78：
In cylinder bottle, add 6-71 (500mg, 1.6117mmol), piperidines-4-alcohol (244.5mg, 2.4175mmol), uses CH
3cN (15mL) dissolves, and finally adds triethylamine (0.67mL, 4.8346mmol), refluxes 72 hours after sealing in 80 DEG C of oil baths.Be spin-dried for, then use dissolve with methanol, add NaHCO
3neutralization, filters.Column chromatography, with triethylamine, ethyl acetate and sherwood oil gradient elution, collect product point, concentrating under reduced pressure, vacuum-drying obtains product 6-78 (541.6mg, productive rate 89.6%).
1H-NMR(400MHz,CDCl
3)δ7.18-7.09(m,2H),7.01(d,J＝8Hz,1H),6.95-6.83(m,4H),3.89(t,J＝6Hz,2H),3.72-3.62(m,1H),2.78-2.68(m,2H),2.46(t,J＝6Hz,2H),2.20-2.06(m,2H),1.98-1.83(m,4H),1.59-1.50(m,2H)。
Embodiment 20 prepares 6-80
In cylinder bottle, add 6-71 (500mg, 1.6117mmol) and dried meat ammonia alcohol (244.5mg, 2.4175mmol), use CH
3cN (15mL) dissolves, and finally adds triethylamine (0.67mL, 4.8346mmol), refluxes 72 hours after sealing in 80 DEG C of oil baths.Stopped reaction, direct evaporate to dryness, then uses dissolve with methanol, adds NaHCO
3neutralization, filters, concentrated.Column chromatography, with ethyl acetate and sherwood oil gradient elution, collect product point, concentrating under reduced pressure vacuum-drying obtains product 6-80 (307.1mg, productive rate 50.8%).
1H-NMR(400MHz,CDCl
3)δ7.20-7.12(m,2H),7.06-7.01(m,1H),6.97-6.92(m,1H),6.92-6.87(m,2H),6.86-6.83(m,1H),3.92(t,J＝6Hz,2H),3.60-3.55(m,1H),3.36-3.31(m,1H),3.19-3.13(m,1H),2.97-2.89(m,1H),2.59-2.52(m,1H),2.40-2.32(m,1H),2.24-2.16(m,1H),2.06-1.90(m,3H),1.87-1.81(m,1H),1.77-1.68(m,3H)。
Embodiment 21 prepares 6-86
6-71 (500mg, 1.6117mmol) is added, Guanidinium hydrochloride (168.4mg in cylinder bottle, 1.7728mmol), with ethanol/water (10mL) dissolved hydrogen sodium oxide (70.9mg, 1.7728mmol), then reflux 72 hours in 80 DEG C of oil baths.Stopped reaction, direct evaporate to dryness.Column chromatography, with ammoniacal liquor, methyl alcohol and dichloromethane gradient drip washing, collect product, concentrating under reduced pressure vacuum-drying obtains product 6-86 (513.9mg, productive rate 100%).
1H-NMR(400MHz,CDCl
3)δ7.23-7.14(m,2H),7.07(d,J＝8Hz,1H),7.00-6.88(m,4H),3.97(t,J＝6Hz,2H),3.28-3.19(m,2H),2.10-2.02(m,2H)。
Embodiment 22 prepares 6-87
6-71 (500mg is added in cylinder bottle, 1.6117mmol), 1,2,4-triazole (222.6mg, 3.2234mmol), dissolve with acetone (5mL), then add salt of wormwood (1.3365g, 9.6701mmol) and potassiumiodide (18.7mg, 0.1612mmol), then reflux 72 hours in 80 DEG C of oil baths.Stopped reaction, direct evaporate to dryness.Column chromatography, with ethyl acetate and sherwood oil gradient elution, collect product, concentrating under reduced pressure, vacuum-drying obtains product 6-87 (552.6mg, productive rate 100%).
1H-NMR(400MHz,CDCl
3)δ7.90(s,1H),7.81(s,1H),7.23-7.14(m,2H),7.10(d,J＝8Hz,1H),7.02-6.91(m,2H),6.84-6.76(m,2H),4.26(t,J＝6Hz,2H),3.78(t,J＝6Hz,2H),2.42-2.34(m,2H)。
Embodiment 23 prepares 6-88
6-71 (500mg is added in cylinder bottle, 1.6117mmol) with tetrazole (225.7mg, 3.2234mmol), dissolve with acetone (5mL), then salt of wormwood (1.3365g is added, 9.6701mmol) with potassiumiodide (18.7mg, 0.1612mmol), then reflux 72 hours in 80 DEG C of oil baths.Stopped reaction, direct evaporate to dryness.Column chromatography, sherwood oil and ethyl acetate gradient drip washing, collect product, concentrating under reduced pressure, vacuum-drying obtains product 6-88 (554.2mg, productive rate 43.09%).
1H-NMR(400MHz,CDCl
3)δ8.47(s,1H),7.21-7.15(m,2H),7.08(d,J＝8Hz,1H),7.01-6.91(m,2H),6.85-6.78(m,2H),4.77(t,J＝6Hz,2H),3.97(t,J＝6Hz,2H),2.55-2.47(m,2H)。
Embodiment 24 prepares 6-89
6-71 (500mg is added in cylinder bottle, 1.6117mmol), 5-methyl isophthalic acid H-tetrazole (270.7mg, 3.2234mmol), dissolve with acetone (5mL), then add salt of wormwood (1.3365g, 9.6701mmol) and potassiumiodide (18.7mg, 0.1612mmol), then reflux 72 hours in 80 DEG C of oil baths.Stopped reaction, direct evaporate to dryness.Column chromatography, with sherwood oil and ethyl acetate gradient drip washing, collect product, concentrating under reduced pressure, vacuum-drying obtains product 6-89 (511.4mg, productive rate 88.67%).
1H-NMR(400MHz,CDCl
3)δ7.20-7.14(m,2H),7.07(d,J＝8Hz,1H),7.00-6.90(m,2H),6.86-6.77(m,2H),4.68(t,J＝6Hz,2H),3.95(t,J＝6Hz,2H),2.51-2.44(m,5H)。:
Embodiment 25 prepares 6-109
6-81：
In cylinder bottle, add 6-71 (500mg, 1.6117mmol), 1-tert-butoxycarbonyl-piperazine (244.5mg, 2.4175mmol), uses CH
3cN (15mL) dissolves, and finally adds triethylamine (0.67mL, 4.8346mmol), seals rapidly, then refluxes 72 hours in 80 DEG C of oil baths.Stopped reaction, direct evaporate to dryness, then uses dissolve with methanol, adds NaHCO
3neutralization, filters, concentrated.Column chromatography, with ethyl acetate and sherwood oil gradient elution, collect product, concentrating under reduced pressure, vacuum-drying obtains product 6-81 (741.4mg, productive rate 100%) ITMS+ESI:[M+H
+] 460.19.
6-109：
In the flask of 50mL, add 6-81 (877.7mg, 1.9079mmol), dissolve (10mL) with methylene dichloride, and then add TFA (0.73mL, 9.5395mmol), stirred overnight at room temperature.Stopped reaction evaporate to dryness, with dissolve with methanol, then adds excessive NaHCO
3neutralize unnecessary TFA, suction filtration, in filtrate, add silica-gel powder concentrate evaporate to dryness, dry method loading column chromatography.With ammoniacal liquor, methyl alcohol and dichloromethane gradient drip washing, collect product point, concentrating under reduced pressure obtains product 6-109 (558.3mg, productive rate 81%).
1H-NMR(400MHz,CDCl
3)δ7.18-7.09(m,2H),7.01(d,J＝8Hz,1H),6.95-6.83(m,4H),3.91(t,J＝6Hz,2H),2.95-2.88(m,4H),2.51-2.42(m,6H),1.97-1.88(m,2H)。
Embodiment 26 prepares 4-219
4-195：
Get 2-chloro phenothiazine (1.00g, 4.2786mmol) and be placed in 100mL tri-neck round-bottomed flask, add through the dried THF (25mL) of Anhydrous potassium carbonate, at N
2add NaH (410.7mg, 17.1145mmol) and the 4-170 (1.2550g, 4.2786mmol) of 60% mineral oil protection under protection, flow through night next time at 60 DEG C of heating conditions.After reaction terminates, stopped reaction, is cooled to room temperature, reaction solution is poured in frozen water (200mL), be extracted with ethyl acetate (200mL × 3), merge organic phase anhydrous magnesium sulfate drying, filter, in filtrate, add silica-gel powder, solid solution is made in drying, dry method loading column chromatography, with ethyl acetate and sherwood oil gradient elution, collect product, evaporate to dryness obtains sterling 4-195 (0.70g, productive rate 47%).ITMS+c ESI：[M+H
+]445.17。4-219：
Get 4-195 (1.50g, 3.1444mmol) and be placed in 100mL flask, add methylene dichloride (50mL), add TFA (2.41mL, 31.4436mmol) under magnetic stirring, overnight at room temperature reacts.Stopped reaction evaporate to dryness, thick product dissolve with methanol, adds excessive NaHCO
3in and TFA, filter, add methylene dichloride and set product dissolved, wet method loading column chromatography.Use NH
3h
2o, methyl alcohol and dichloromethane gradient drip washing, collect product, and concentrated, drying obtains product 4-219 (654.4mg, productive rate 60%).
1H-NMR(500MHz,CDCl
3)δ7.20-7.165(m,2H),7.09-7.055(m,1H),7.00-6.95(m,1H),6.95-6.915(dd,1H),6.91-6.87(d,1H),6.865-6.85(d,1H),4.00-3.94(t,2H),3.81-3.42(m,2H),2.975-2.9425(s,1H),2.9025-2.8675(s,1H)2.26-2.12(m,1H),2.005-1.8975(m,1H),1.768-1.665(m,2H),1.635-1.505(m,2H),1.495-1.40(m,2H)；ITMS+c ESI：[M+H
+]345.22。
Bioassay and result
This patent has carried out detailed test for MCF-7 cell strainHJ2mm and Human hepatoma cell line Hep-G2 to the cytotoxicity of synthesized compound.MCF-7 and Hep-G2 cell strain substratum used for containing 10%NCS, 1% dual anti-DMEM-F
12, insolution.Culturing cell, to its exponential phase of growth, uses cell counting count board to count cell suspension, uses nutrient solution to regulate cell concn to be 20000/mL, then by cell suspension inoculation in 96 orifice plates, 100 μ L are inoculated in every hole, and reserve 4 holes as blank, then culture plate are positioned over 5%CO
2, cultivate in 37 DEG C of incubators and make cell attachment in 24 hours.Take out culture plate nutrient solution is wherein removed, then add each concentration successively and (take each compound in the desired amount, then each compound is first dissolved with 50 μ lDMSO, then be diluted to 100 μm of ol/L, 50 μm of ol/L, 25 μm of ol/L, 12.5 μm of ol/L, 6.25 μm of ol/L, 3.125 μm of ol/L, 1.5625 μm of ol/L successively with blank nutrient solution) the nutrient solution of pastille, blank control wells adds blank nutrient solution, puts into incubator and continues cultivation 72 hours.The compound that known anticancer drugs, doxorubicin, taxol are prepared with the present invention is respectively made positive control, takes out culture plate, add the MTT reagent (20 μ L/ hole) of 5mg/mL in respective aperture respectively, then culture plate is relay people 5%CO
2, continue cultivation 4 hours in 37 DEG C of incubators, take out culture plate, the nutrient solution in porose for institute removed, then adds DMSO (150 μ L/ hole), use enzyme-linked immunosorbent assay instrument in the OD value of wavelength 490nm place measurement cell, then use IC
50software for calculation calculates the IC of each compound
50value, as shown in table 2.According to the present invention, not expected discovery, the compound that the present invention prepares gained all can produce certain restraining effect for MCF-7 and Hep-G2 cell strain.
Table 2 phenothiazine compound IC
50value
Claims (10)
1. a phenothiazine compound, is characterized in that, this compound structure is as shown in logical formula I:
Wherein, A is piperazinyl, piperidyl, triazol radical, tetrazole base, pyrryl, guanidine radicals, morpholinyl and derivatives group thereof; R is C
2-3straight chain saturated alkyl; B can be independently Cl, methylthio group or trifluoromethyl.
2. phenothiazine compound according to claim 1, is characterized in that, when B is methylthio group, A is piperazinyl, triazol radical, tetrazole base, pyrryl, guanidine radicals, morpholinyl and derivatives group thereof, or piperidyl or N (CH
3)
2; When B is trifluoromethyl, A is morpholinyl, piperidyl, pyrryl and derivatives group thereof, or piperazinyl; When B is Cl, A is piperidyl, triazol radical, tetrazole base, pyrryl, guanidine radicals, morpholinyl and derivatives group thereof, or piperazinyl.
3. the preparation method of phenothiazine compound according to claim 1, is characterized in that, presoma has the chemical structure of logical formula II, and the T1 group of this presoma and ligand groups in organic solvent substitution reaction occur; Wherein T
1be selected from halogenopropane and derivatives group thereof, T
2be selected from Cl, methylthio group or trifluoromethyl;
Described ligand groups comprises: piperidyl, piperazinyl, triazol radical, tetrazole base, pyrryl, guanidine radicals, morpholinyl and derivatives group thereof.
4. the preparation method of phenothiazine compound according to claim 1, is characterized in that, M in organic solvent substitution reaction occurs with the compound with logical formula III;
Wherein, M be selected from the halothane that replaces by piperidyl or derivatives thereof group and derivative thereof, B can be independently Cl, methylthio group or trifluoromethyl.
5. the preparation method according to claim 3 or 4, is characterized in that, described general formula (I) or the piperidyl of (II), piperazinyl and derivatives group thereof are connected by amidation or esterification by one or continuous two amino acid.
6. the preparation method according to claim 3 or 4, it is characterized in that, described organic solvent as the mixture of a kind of or above-mentioned solvent in ether, alkane, aromatic hydrocarbon, amides, nitrile, amine, alcohols or ketones solvent, as THF, toluene, DMF, acetonitrile, triethylamine, acetone or alcohol.
7. the application of phenothiazine compound according to claim 1, wherein said phenothiazine compound, for the preparation of the medicine of anti-human breast cancer cell line mcf-7.
8. the application of phenothiazine compound according to claim 1, wherein said phenothiazine compound, for the preparation of the medicine of anti-human liver cancer cell strain Hep-G2.
9. an anticancer pharmaceutical composition, the arbitrary compound wherein containing claim 1 ~ 2 or their mixture are as effective constituent, and one or more pharmaceutically acceptable carriers.
10. the application of phenothiazine compound according to claim 9, wherein pharmaceutical composition can make the pharmaceutically acceptable formulation of any one, injection, sprays, inhalation or oral preparation.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510271507.5A CN104829554B (en) | 2015-05-25 | 2015-05-25 | Phenothiazine compound and its preparation method and application |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510271507.5A CN104829554B (en) | 2015-05-25 | 2015-05-25 | Phenothiazine compound and its preparation method and application |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN104829554A true CN104829554A (en) | 2015-08-12 |
| CN104829554B CN104829554B (en) | 2018-07-13 |
Family
ID=53807797
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201510271507.5A Expired - Fee Related CN104829554B (en) | 2015-05-25 | 2015-05-25 | Phenothiazine compound and its preparation method and application |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104829554B (en) |
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108530384A (en) * | 2018-03-26 | 2018-09-14 | 南京泽恒医药技术开发有限公司 | A kind of preparation method of prochlorperazine |
| CN109761932A (en) * | 2019-01-29 | 2019-05-17 | 天津国际生物医药联合研究院 | Phenothiazine compound and its application |
| EP3574920A1 (en) * | 2018-05-31 | 2019-12-04 | Institut National De La Sante Et De La Recherche Medicale (Inserm) | Nupr1 inhibition for treating cancer |
| RU2732417C1 (en) * | 2016-10-17 | 2020-09-16 | Энрок Фармасьютикал Текнолоджиз (Хэбэй) Лимитед | Phenothiazine derivatives and methods for use thereof |
| IT201900015809A1 (en) * | 2019-09-06 | 2021-03-06 | Univ Degli Studi Padova | COMPOUNDS SIMILAR TO THYORIDAZINE |
| CN113264903A (en) * | 2021-05-27 | 2021-08-17 | 郑州大学 | Phenothiazine compound and preparation method and application thereof |
| EP3773597A4 (en) * | 2018-04-06 | 2022-06-08 | Camp4 Therapeutics Corporation | Phenothiazine derivatives and uses thereof |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102516200A (en) * | 2011-12-14 | 2012-06-27 | 合肥工业大学 | Hybrid compound with indanone and phenothiazine as skeletal structures and application thereof |
| CN104114175A (en) * | 2011-10-28 | 2014-10-22 | 黄奇英 | Pharmaceutical composition for elimination of cancer stem cells |
-
2015
- 2015-05-25 CN CN201510271507.5A patent/CN104829554B/en not_active Expired - Fee Related
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104114175A (en) * | 2011-10-28 | 2014-10-22 | 黄奇英 | Pharmaceutical composition for elimination of cancer stem cells |
| CN102516200A (en) * | 2011-12-14 | 2012-06-27 | 合肥工业大学 | Hybrid compound with indanone and phenothiazine as skeletal structures and application thereof |
Non-Patent Citations (2)
| Title |
|---|
| 孟树林等: "硫利达嗪对肝癌干细胞的杀伤作用研究", 《中国生物工程杂志》 * |
| 童斯浩等: "硫利达嗪对乳腺癌细胞MDA-MB-231和MCF-7的杀伤效应", 《安徽医科大学学报》 * |
Cited By (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| RU2732417C1 (en) * | 2016-10-17 | 2020-09-16 | Энрок Фармасьютикал Текнолоджиз (Хэбэй) Лимитед | Phenothiazine derivatives and methods for use thereof |
| CN108530384A (en) * | 2018-03-26 | 2018-09-14 | 南京泽恒医药技术开发有限公司 | A kind of preparation method of prochlorperazine |
| CN108530384B (en) * | 2018-03-26 | 2021-12-28 | 南京泽恒医药技术开发有限公司 | Preparation method of prochlorperazine |
| EP3773597A4 (en) * | 2018-04-06 | 2022-06-08 | Camp4 Therapeutics Corporation | Phenothiazine derivatives and uses thereof |
| US11542290B2 (en) | 2018-04-06 | 2023-01-03 | Camp4 Therapeutics Corporation | Phenothiazine derivatives and uses thereof |
| EP3574920A1 (en) * | 2018-05-31 | 2019-12-04 | Institut National De La Sante Et De La Recherche Medicale (Inserm) | Nupr1 inhibition for treating cancer |
| WO2019229236A1 (en) * | 2018-05-31 | 2019-12-05 | Institut National De La Sante Et De La Recherche Medicale (Inserm) | Nupr1 inhibition for treating cancer |
| CN109761932A (en) * | 2019-01-29 | 2019-05-17 | 天津国际生物医药联合研究院 | Phenothiazine compound and its application |
| IT201900015809A1 (en) * | 2019-09-06 | 2021-03-06 | Univ Degli Studi Padova | COMPOUNDS SIMILAR TO THYORIDAZINE |
| WO2021043862A1 (en) * | 2019-09-06 | 2021-03-11 | Universita' Degli Studi Di Padova | Thioridazine derivatives and their use for the treatment of cancer |
| CN113264903A (en) * | 2021-05-27 | 2021-08-17 | 郑州大学 | Phenothiazine compound and preparation method and application thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN104829554B (en) | 2018-07-13 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN104829554A (en) | Phenothiazine compound and preparation method and application thereof | |
| EP3507284A1 (en) | Biaryl compounds useful as immunomodulators | |
| EP2124927A2 (en) | Carbamate compounds | |
| EP3111940B1 (en) | Silicon phthalocyanine complex, preparation method and medicinal application thereof | |
| CN109180681B (en) | DNA toxic dimer compound | |
| EP3086792B1 (en) | Methods and reagents for radiolabeling | |
| CN106977415B (en) | Intermediate of shakubiqu and preparation method thereof | |
| CN108218798B (en) | Preparation method of Apabetalone | |
| CN104086514A (en) | Paclitaxel derivatives and preparation method thereof | |
| WO2014056083A1 (en) | Kinase inhibitors and method of treating cancer with same | |
| CN114716444B (en) | Chlorin e6-DCA conjugate, and preparation method and application thereof | |
| MXPA01013340A (en) | Novel xanthone compounds, their preparation and use as medicament. | |
| CN106946974B (en) | Ursolic amide derivative containing pyrazole heterocycle and synthesis and application thereof | |
| CN108066340B (en) | Pharmaceutical composition | |
| CN112175014A (en) | Nitric oxide donor type tetravalent platinum derivative, preparation method and medical application thereof | |
| CN110483405B (en) | Kealiinine derivatives, preparation thereof and application thereof in resisting plant viruses and germs | |
| CN106146497A (en) | Matrine oxime ester derivative and preparation method and application | |
| CN111018780B (en) | N-carbonyl-9, 10-dihydroacridine compound and application thereof | |
| CN107325030A (en) | The new trans-stilbene class antitumor agent of one class | |
| CN109867708A (en) | The preparation method and application of enoxolone series derivates (TNGA-X) with antitumor action | |
| CN104341407A (en) | Quinazoline compounds, preparation method and applications thereof | |
| CA2423454C (en) | Estrogen-linked platinum (ii) complexes as anticancer agents | |
| RU2748533C2 (en) | New pheospheride derivatives with antitumor activity, method for obtaining and application of these compounds | |
| CN107382941B (en) | Flavone derivative and preparation method and application thereof | |
| CN108069964B (en) | Macrocyclic compounds and pharmaceutical compositions |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| EXSB | Decision made by sipo to initiate substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20180713 Termination date: 20210525 |
|
| CF01 | Termination of patent right due to non-payment of annual fee |