CN111793035B - N4- (3-methoxyphenyl) -pyrimidinediamine targeted DDR1 inhibitor and preparation and application thereof - Google Patents
N4- (3-methoxyphenyl) -pyrimidinediamine targeted DDR1 inhibitor and preparation and application thereof Download PDFInfo
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Abstract
The invention discloses a method for preparing N4- (3-methoxyphenyl) -pyrimidinediamine targeted DDR1 inhibitor and preparation method and application thereof. N of the invention4The- (3-methoxyphenyl) -pyrimidinediamine targeted DDR1 inhibitor shows that the inhibitor can relieve non-small cell lung cancer gefitinib resistant cell PC-9G, and has certain antitumor activity on other 3 strains of DDR1 high-expression cells. According to the result of the antitumor activity test, the compound D06 shows the biological activity superior to that of the primer Yfq07 in PC-9G cells; and shows better activity (IC) to A549, A431 and HCT116 highly expressed by DDR1501.20-2.53 μ M), and is close to or better than the lead agent Yfq 07; DDR1 kinase test results show that the inhibition rate reaches 76.2% at 10 muM concentration, and the selectivity is more than 3 times higher than that of the primer Yfq 07; meanwhile, Western blot experiments prove that the compound D06 can inhibit the phosphorylation of DDR1 in PC-9G cells.
Description
Technical Field
The invention relates to the technical field of medical chemistry, in particular to N4- (3-methoxyphenyl) -N6The-substituted pyrimidine-4, 6-diamine targeting DDR1 can relieve PC-9G cell compounds, and a preparation method and an application of the anti-tumor activity thereof.
Background
Discoidin Domain Receptors (DDR) are members of the transmembrane Receptor Tyrosine Kinase (RTK) superfamily discovered in the early 1990 s and are distinguished from other RTKs by the presence of a discoidin motif in the extracellular domain. Currently, typical RTKs use peptide-like growth factors as ligands, but DDR can be activated by collagen, the most abundant component of the extracellular matrix (ECM). Researchers have identified two types of DDR, including DDR1 and DDR 2. Among them, DDR1 can bind to almost all types of collagen reported so far; meanwhile, DDR1 is widely expressed in epithelial cells of lung, kidney, colon and brain; plays an important role in the regulation of essential cellular processes, including proliferation, survival, differentiation, adhesion and matrix remodeling, and is closely associated with many human diseases, including various cancers, fibrotic diseases and atherosclerosis, among others.
Compound Yfq07 was originally independently developed by this unit to design synthetic small molecule inhibitors targeting EGFR. In the process of further research on the subsequent biological activity, the proteomic experiment (as shown in fig. 1) shows that the expression level of EGFR is remarkably reduced in gefitinib-resistant PC-9G cells, while the expression level of DDR1 is abnormally increased, and the expression of DDR1 is remarkably inhibited in gefitinib-resistant PC-9G cells added with compound Yfq 07. Therefore, we speculate that activation of the DDR1 alternative signaling pathway may occur at the same time as the EGFR pathway is blocked in gefitinib-resistant PC-9G cells. The compound Yfq07 can obviously reduce the expression of DDR1, and further has a certain inhibition effect on PC-9G cells. After a large number of literature queries, DDR1 was found to activate as a gefitinib-resistant alternative signal in non-small cell lung cancer, rarely reported. Therefore, to further verify the new finding, Yfq07 is selected as a lead to carry out structural modification so as to obtain the antitumor drug with higher targeting DDR1 selectivity and better activity.
Disclosure of Invention
The invention provides N4- (3-methoxyphenyl) -pyrimidinediamine targeted DDR1 inhibitor, preparation and application thereof, and N4Compared with the prior art, the- (3-methoxyphenyl) -pyrimidinediamine targeted DDR1 inhibitor has better antitumor activity.
N4- (3-methoxyphenyl) -pyrimidinediamine targeted DDR1 inhibitor, the structural formula is as follows:
wherein R is a substituted or unsubstituted benzene or heteroaromatic ring;
the substituent on the benzene ring or the heteroaromatic ring is one or more of halogen, methyl, methoxy, trifluoromethyl and nitro.
Preferably, N is4The- (3-methoxyphenyl) -pyrimidinediamine targeted DDR1 inhibitor is one of compounds D01-06 and Z07, and R of the compounds D01-06 and Z07 is shown in Table 1:
most preferably, said N4The- (3-methoxyphenyl) -pyrimidinediamine targeted DDR1 inhibitor is a compound N4- (3-methoxyphenyl) -N6- (4- ((4-methylpiperazin-1-yl) methyl) -3- (trifluoromethyl) phenyl) pyrimidine-4, 6-diamine (D06), having the chemical structure:
the invention also provides the N4A method of targeting DDR1 inhibitors of the- (3-methoxyphenyl) -pyrimidinediamines, comprising the steps of:
(1)4, 6-dichloropyrimidine and 3-methoxyaniline react under the action of DIPEA to obtain an intermediate 6-chloro-N- (3-methoxyphenyl) pyrimidine-4-amine;
(2) the intermediate 6-chloro-N- (3-methoxyphenyl) pyrimidine-4-amine and 4- ((4-methylpiperazin-1-yl) methyl) -3- (trifluoromethyl) aniline react under the action of alkali, ligand and palladium catalyst to obtain the N4- (3-methoxyphenyl) -pyrimidinediamines target DDR1 inhibitors.
The preparation method specifically comprises the following steps:
(1) the starting materials 4, 6-dichloropyrimidine (1.8g,12.1mmol) and 3-methoxyaniline (1g, 8.1mmol) were weighed out, added to a 100mL single-neck round-bottom flask, and dissolved in 20mL absolute ethanol. After the mixture was sufficiently dissolved by sonication, DIPEA (1.6mL,9.7mmol) was added dropwise, and the mixture was heated under reflux at 80 ℃ for 8 to 12 hours, and the progress of the reaction was monitored by TLC. After the reaction is finished, adding a proper amount of column chromatography silica gel, spinning dry to prepare sand, separating and purifying by column chromatography, drying and weighing to obtain 1.5g of intermediate 6-chloro-N- (3-methoxyphenyl) pyrimidine-4-amine, wherein the yield is 79.8 percent, and the melting point is 153.3-155.6 ℃;
(2) a100 mL dry three-necked round bottom flask was taken and charged with the intermediates 6-chloro-N- (3-methoxyphenyl) pyrimidin-4-amine (280mg,1.2mmol), 4- ((4-methylpiperazin-1-yl) methyl) -3- (trifluoromethyl) aniline (273mg,1mmol), cesium carbonate (490mg,1.5mmol), Xantphos (25mg,0.04mmol), Pd in that order2(dba)3(20mg,0.02 mmol). Under the protection of nitrogen, 20mL of anhydrous dioxane is added, heating and refluxing are carried out for 12-24 h at 95 ℃, and the reaction process is monitored by a TLC method. After the reaction is finished, adding a proper amount of column chromatography silica gel, concentrating the reaction solution under reduced pressure, spinning to prepare sand, separating and purifying by column chromatography, drying and weighing. Through ESI-HRMS, the method is carried out,1H-NMR,13C-NMR, UPLC identification results; obtaining a target compound with a melting point of 217.6-219.0 ℃ and a yield of 28.37%;
the invention also provides the N4- (3-methoxyphenyl) -pyrimidinediamines targeting DDR1 inhibitionThe application of the preparation in preparing antitumor drugs for treating tumors.
Preferably, the antitumor drug is used for treating lung cancer; most preferably, the lung cancer is non-small cell lung cancer.
N of the invention4The- (3-methoxyphenyl) -substituted pyrimidinediamines show certain antitumor activity. According to the result of the antitumor activity test, the compound D06 shows a biological activity superior to that of the lead drug Yfq 07; and shows excellent activity (IC) on A549, A431 and HCT116 highly expressed by DDR1501.20 to 2.53 μ M); the DDR1 kinase activity test also shows higher selectivity; the western blot experiment proves that D06 can inhibit the phosphorylation of DDR1 in PC-9G cells.
Drawings
FIG. 1 shows quantitative information on phosphorylation sites of differential proteins.
FIG. 2 shows the inhibition of DDR1 phosphorylation in PC-9G cells by compound D06.
Detailed Description
The following examples are further detailed descriptions of the present invention.
Synthesis of the Compound of example 1
1.1A specific synthetic route for the compounds is shown below:
the synthetic routes of the compounds D01-06 and Z07 are as follows: a is CH3CH2OH,DIPEA,80℃,8～12 h；b:1,4-Dioxane,Cs2CO3,Pd2(dba)3,Xantphos,95℃,N2,12-24h
1.2 synthetic procedure
Synthesis of Compounds D01-06 and Z07
a. The starting materials 4, 6-dichloropyrimidine (1.8g,12.1mmol) and 3-methoxyaniline (1g, 8.1mmol) were weighed out, added to a 100mL single-neck round-bottom flask, and dissolved in 20mL absolute ethanol. After the mixture is fully dissolved by ultrasonic treatment, DIPEA (1.6mL,9.7mmol) is added dropwise, the mixture is heated and refluxed for 8-12 h at 80 ℃, and the reaction process is monitored by a TLC method. After the reaction is finished, adding a proper amount of column chromatography silica gel, spinning dry to prepare sand, separating and purifying by column chromatography, drying and weighing to obtain 1.5g of intermediate 6-chloro-N- (3-methoxyphenyl) pyrimidine-4-amine, wherein the yield is 79.8 percent, and the melting point is 153.3-155.6 ℃;
b. a100 mL dry three-necked round bottom flask was taken and charged with intermediates 6-chloro-N- (3-methoxyphenyl) pyrimidin-4-amine (280mg,1.2mmol), heterocycle substituted aniline (1mmol), cesium carbonate (490mg,1.5mmol), Xantphos (25mg,0.04mmol), Pd in that order2(dba)3(20mg,0.02 mmol). Under the protection of nitrogen, 20mL of anhydrous dioxane is added, heating and refluxing are carried out for 12-24 h at 95 ℃, and the reaction process is monitored by a TLC method. After the reaction is finished, adding a proper amount of column chromatography silica gel, decompressing and concentrating the reaction solution, spinning and drying to prepare sand, separating and purifying by column chromatography, drying and weighing, calculating the yield, measuring the melting point, carrying out ESI-HRMS,1H-NMR,13C-NMR, UPLC identification results;
1.3 results of the experiment
All the target compound structures synthesized are shown in the table 1 above; ESI-HRMS of a portion of the target compound synthesized including the active compound,1H-NMR,13C-NMR, UPLC and other physical and chemical data are as follows:
N4-(3-methoxyphenyl)-N6-(quinolin-3-yl)pyrimidine-4,6-diamine(D 01)
Chemical Formula:C20H17N5O；Yield/％:57.61％；MP:203.5～205.8℃；UPLC:Purity: 100.0％；HRMS(ESI)for C20H17N5O[M+H]+,calcd:343.1442.Found:343.1438.；1H-NMR(500MHz,DMSO-d6)δ(ppm):9.71(s,1H,Ar-H),9.29(s,1H,Ar-H),8.94(s, 1H,-NH-),8.74(s,1H,Ar-H),8.41(s,1H,Ph-H),7.93(d,1H,J＝8.0Hz,Ar-H),7.88(d,1H, J＝7.5Hz,Ar-H),7.56(m,2H,Ar-H),7.23(m,2H,-NH-+Ar-H),7.14(d,1H,J＝7.6Hz, Ph-H),6.59(d,1H,J＝7.6Hz,Ph-H),6.32(s,1H,Ar-H),3.75(s,3H,-OCH3)；13C-NMR (125MHz,DMSO-d6):160.5,160.3,159.7,157.7,145.4,143.2,141.4,134.5,129.5,128.4, 128.1,127.2,126.9,120.8,112.3,107.3,105.9,87.5,59.7,55.0；
N4-(3-methoxyphenyl)-N6-(quinazolin-4-yl)pyrimidine-4,6-diamine(D 03)
Chemical Formula:C19H16N6O；Yield/％:65.98％；MP:206.4～209.7℃；UPLC:Purity: 100.0％；HRMS(ESI)for C19H16N6O[M+H]+,calcd:344.1446.Found:344.1441.；1H-NMR(500 MHz,DMSO-d6)δ(ppm):10.37(s,1H,Ar-H),9.67(s,1H,-NH-),8.35(s, 1H,Ar-H),8.75(d,1H,J＝8.2 Hz,Ar-H),8.51(s,1H,Ph-H),8.10(s,1H,-NH-),7.91(m, 2H,Ar-H),7.66(t,1H,J＝7.5 Hz,Ar-H),7.44(s,1H,Ar-H),7.25(m,2H,Ph-H),6.60(d, 1H,J＝7.0 Hz,Ph-H),3.77(s,3H,-OCH3)；13C-NMR(125 MHz,DMSO-d6):161.7,161.1, 156.3,152.8,150.6,149.8,143.4,132.0,130.5,128.8,127.8,126.4,116.2,110.8,110.1, 108.5,99.4,82.6,55.8；
N4-(3-methoxyphenyl)-N6-(4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phen yl)pyrimidine-4,6-diamine(D 06)
Chemical Formula:C24H27F3N6O；Yield/％:28.37％；MP:217.6～219.0℃；UPLC:Purity: 90.8％；HRMS(ESI)for C24H27F3N6O[M+H]+,calcd:472.2254.Found:472.2249.；1H-NMR(500 MHz,DMSO-d6)δ(ppm):9.54(s,1H,Ar-H),9.26(s,1H,Ph-H),8.33(s, 1H,Ph-H),8.05(s,1H,-NH-),7.86(d,1H,J＝8.0 Hz,Ph-H),7.60(d,1H,J＝8.5 Hz,Ph-H), 7.20(m,2H,-NH-+Ar-H),7.12(d,1H,J＝8.0 Hz,Ph-H),6.57(d,1H,J＝7.0 Hz,Ph-H), 6.22(s,1H,Ar-H),3.74(s,3H,Ph-H),3.53(s,2H,-CH2-),2.43(br s,8H),2.24(s,3H, -CH3)；13C-NMR(125 MHz,DMSO-d6):160.5,160.2,159.7,157.6,141.4,139.9,131.3, 129.4,122.3,112.3,107.2,105.8,87.3,57.3,54.9,54.4,52.1,45.1；
N4-(2,5-dimethoxypyrimidin-4-yl)-N6-(3-methoxyphenyl)pyrimidine-4,6-diamine (Z 07)
Chemical Formula:C17H18N6O3；Yield/％:67.45％；MP:197.4～199.0℃；UPLC:Purity: 100.0％；HRMS(ESI)for C17H18N6O3[M+H]+,calcd:354.1447.Found:354.1439.；1H-NMR(500 MHz,DMSO-d6)δ(ppm):9.66(s,1H,-NH-),8.38(s,1H,Ar-H),8.29(s, 1H,Ar-H),8.03(s,1H,Ph-H),7.74(s,1H,-NH-),7.33(s,1H,Ar-H),7.22(m,2H,Ph-H), 6.60(m,1H,J＝3.0 Hz,Ph-H),3.90(s,3H,-OCH3),3.82(s,3H,-OCH3),3.75(s,3H, -OCH3)；13C-NMR(125MHz,DMSO-d6):161.4,159.6,158.1,157.5,156.2,151.2,141.1, 137.4,136.6,129.4,112.6,107.8,106.2,92.1,57.1,54.9,54.3；
the properties and solubility of the target compound synthesized by the present invention are as follows:
the yield of the target compound is ideal, most of the target compound can reach 60-90%, and the yield of individual compounds such as D06 is only 20-30%; the color is mainly white or yellowish. The polarity of the compound is generally large, all the compounds are insoluble in solvents with small polarity such as petroleum ether and n-hexane, and part of the compounds are slightly soluble in ethyl acetate; except that D04 is insoluble in DMSO, DCM and MeOH, the rest of the target compounds can be soluble in DMSO, DCM and the like.
The target compound synthesized by the invention has the following mass spectrum results:all compounds can be seen in [ M + H ]]+Molecular ion peak, wherein most of the compounds are removed by [ M + H ]]+1/2[ M + H ] with higher abundance can be seen in addition to the molecular ion peak of (A)]+An ion peak;1H-NMR spectrum results show that all compound hydrogen numbers, corresponding chemical shifts, coupling constants and the like can be consistent with theoretical values of corresponding compounds;13C-NMR spectrum results show that all compound carbon peak shifts and numbers accord with theoretical data;
EXAMPLE 2 antitumor cell Activity of Compounds
2.1 testing the antitumor Activity of Compounds by the MTT method
In the experiment, the cell survival rate of the non-small cell lung cancer gefitinib resistant cell PC-9G, the non-small cell lung cancer cell A549, the human epidermal carcinoma cell A431 and the colon cancer cell HCT-116 is detected by an MTT method. Each tumor cell (PC-9G, A549, A431 and HCT-116) in logarithmic growth phase was cultured separately in 96-well plates at a plating concentration of about 5X 10 per well3One cell filled with 5.0% CO2Culturing for 24 hours at the constant temperature of 37 ℃; observed under a microscope, the final concentration dissolved in DMSO is given after the cells are attached: lead compound Yfq07 with different concentrations and target compound D06 with better activity were dissolved in 1. mu.L. After 48h of administration and culture, 20. mu.L of MTT solution dissolved in PBS at 5mg/mL was added to each well and culture was continued for 4h, and formazan precipitate visible to the naked eye was observed to be formed; the solution in each well was carefully discarded and 150 μ L DMSO was added to each well for dissolving formazan crystals and uniformly shaken on a shaker for 10 min; finally, detecting the light absorption value of ultraviolet absorption wavelength at 490nm of each hole by using a microplate reader, and calculating the corresponding cell survival rate, inhibition rate and IC (integrated circuit) through conversion50The value is obtained. This experiment requires at least three replicates.
2.2 results of the experiment
IC of the screened active compound D06 and the pilot Yfq07 on the non-small cell lung cancer gefitinib resistant cell PC-9G, the non-small cell lung cancer cell A549, the human epidermal carcinoma cell A431 and the colon cancer cell HCT-116 which are highly expressed in DDR1 by an MTT method50The values are determined, and the corresponding experimental results are shown in table 2;
table 2: IC of active Compound D0650(μM)
The results show that: the compound D06 shows better inhibition effect in A549 cells, A431 cells and HCT116 cells, and IC50The value is between 1.20 and 2.53 mu M; furthermore, IC in PC-9G cells50The value is 0.74 +/-0.03 mu M, which is superior to the leader Yfq 07.
Example 3 Compounds Activity against DDR1 kinase
3.1 DDR1 kinase assay
In this experiment, the inhibition effect of the target compound on DDR1 kinase was evaluated by performing a kinase reaction in a 10 μ L small-volume 384-well plate using the lantha screen Eu kinase activity assay technique. The kinase in reaction buffer was composed of 50mM HEPES pH 7.5, 0.01% BRIJ-35, 10mM MgCl2And 1mM EGTA, at a concentration of 100nM for the fluorescein-poly GAT substrate (Invitrogen, USA). The kinase reaction was initiated by the addition of 100nM ATP in the presence of a series of dilutions. The reaction was allowed to proceed at room temperature for 1 hour, then 10. mu.L of EDTA (20mM) and Eu-labeled antibody (4nM) in TR-FRET dilution buffer were added. The final concentration of antibody in the assay wells was 2nM and EDTA 10 mM. The plates were incubated for an additional hour at room temperature and then a TR-FRET emission ratio of 665 nm/340nm was obtained on a Perkin Elmer EnVision MultiMark reader (Perkin-Elmer, Inc.). Curve fitting and data analysis were performed using GraphPad Prism4 software.
3.2 results of the experiment
Note: yfq07 is a lead compound. All compounds tested were at 10 μ M and at least three experiments were performed for each compound.
The results show that: (1) the DDR1 kinase inhibition rate of the lead compound Yfq07 is 22.1%; (2) in the target compounds, the inhibition rates of D01, D04 and D06 on DDR1 kinase are superior to those of lead compound Yfq 07; (3) the compound D06 is the target compound with the best DDR1 kinase inhibitory activity, and the DDR1 kinase inhibitory rate reaches 76.2%.
EXAMPLE 4 inhibition of DDR1 in PC-9G cells by Compound D06
4.1 immunoblotting of DDR1 phosphorylation inhibition
In the experiment, the Western blot method is adopted to detect the target compound with the best activity on the protein level. Non-small cell lung cancer gefitinib resistant cell PC-9G is paved in a 6-well plate and filled with 5.0% CO at constant temperature of 37 DEG C2Starving and culturing for 24 hours under the condition; the next day, 0 μ M, 2.5 μ M, 5 μ M, 10 μ M of the active compound to be tested, 10 μ M of lead compound Yfq07 were given for further culture for 24 h; collecting cells, washing with PBS three times, adding in-situ lysate (total protein extraction reagent: phosphatase inhibitor: PMSF ═ 100:1:1), placing on ice for lysis for 10min, scraping and collecting protein; centrifuging the collected protein by a high-speed low-temperature centrifuge at-4 deg.C and 12000r/min for 20min, and collecting supernatant to clean EP tube. Calculating the concentration of the required protein through a standard protein curve, loading, carrying out SDS-PAGE electrophoresis, transferring the protein onto a PDVF membrane, sealing the PDVF membrane by using skimmed milk, washing the PDVF membrane by using a TBST solution, putting the cleaned PDVF membrane into a corresponding primary antibody, and incubating the primary antibody in a refrigerator at 4 ℃ overnight; the next day, primary antibodies are recovered and labeled, and corresponding secondary antibodies are incubated at the same time; and finally, adding exposure liquid, and detecting by using an exposure instrument.
4.2 results of the experiment
In order to explore the effect of an active compound D06 on DDR1 protein, non-small cell lung cancer gefitinib-resistant PC-9G cells were selected, and Western blot experiments were used to detect the inhibition effect of the active compound D06 on DDR1 protein and DDR1 phosphorylation at different concentrations, as shown in FIG. 2, FIG. 2 is the inhibition effect of DDR1 phosphorylation of the compound D06 at different concentrations, wherein Yfq07 is a lead compound.
The results show that: the active compound D06 shows obvious DDR1 phosphorylation inhibition effect at 2.5 mu M, and the effect is stronger than that of the lead compound Yfq 07. In addition, D06 has obvious inhibition effect on the total amount of DDR1 protein at 10 mu M, and the inhibition effect is better than that of a positive control at the same concentration.
Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that changes, modifications, substitutions and alterations can be made in these embodiments without departing from the principles and spirit of the invention, the scope of which is defined in the appended claims and their equivalents.
Claims (6)
1. A kind ofN 4- (3-methoxyphenyl) -pyrimidinediamine targeted DDR1 inhibitors characterized by being compound D04 or compound D06;
the structural formula of compound D04 or D06 is as follows:
r is defined as follows:
。
2. The method of claim 1N 4The- (3-methoxyphenyl) -pyrimidinediamine targeted DDR1 inhibitor is characterized by being a compoundN 4- (3-methoxyphenyl) -N 6- (4- ((4-methylpiperazin-1-yl) methyl) -3- (trifluoromethyl) phenyl) pyrimidine-4, 6-diamine (D06), having the chemical structure:
3. a process for preparing a compound according to claim 2N 4A method for targeting DDR1 inhibitors with (3-methoxyphenyl) -pyrimidinediamines, comprising the steps of:
(1)4, 6-dichloropyrimidine and 3-methoxyaniline react under the action of DIPEA to obtain an intermediate 6-chloro-N- (3-methoxyphenyl) pyrimidin-4-amine;
(2) intermediate 6-chloro-NReacting- (3-methoxyphenyl) pyrimidine-4-amine with 4- ((4-methylpiperazin-1-yl) methyl) -3- (trifluoromethyl) aniline under the action of alkali, ligand and palladium catalyst to obtain the compoundN 4- (3-methoxyphenyl) -pyrimidinediamines target DDR1 inhibitors.
4. A process as claimed in any one of claims 1 to 2N 4The application of the- (3-methoxyphenyl) -pyrimidinediamine targeted DDR1 inhibitor in the preparation of antitumor drugs is characterized in that the antitumor drugs are used for treating tumors.
5. The method of claim 4N 4The application of the- (3-methoxyphenyl) -pyrimidinediamine targeted DDR1 inhibitor in the preparation of antitumor drugs is characterized in that the antitumor drugs are used for treating lung cancer.
6. According to the rightThe method according to claim 5N 4The application of the- (3-methoxyphenyl) -pyrimidinediamine targeted DDR1 inhibitor in the preparation of antitumor drugs is characterized in that the lung cancer is non-small cell lung cancer.
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| CN104803925A (en) * | 2015-04-16 | 2015-07-29 | 温州医科大学 | 2,4,5-trisubstituted pyrimidine compounds taking FGFRs (fibroblast growth factor receptors) as targets as well as preparation methods and application of 2,4,5-trisubstituted pyrimidine compounds |
| CN104844526A (en) * | 2015-04-16 | 2015-08-19 | 温州医科大学 | 4,6-pyrimidine diamine compound and preparing method and application thereof |
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