CN103251600B - Antitumor application of 2-amino-4-(3'-cyano-4'-pyrrolidyl)phenylpyrimidine compound - Google Patents

Abstract

The invention discloses an antitumor application of a 2-amino-4-(3'-cyano-4'-pyrrolidyl)phenylpyrimidine compound. Researches show that the compound is capable of doubly inhibiting TBK1 (TANK-binding kinase-1) and IKKi in a targeted manner, and has better proliferation inhibition potentials in a plurality of tumor cell strains cultured in vitro; and experiments prove that 2-amino-4-(3'-cyano-4'-pyrrolidyl)phenylpyrimidine is capable of down-regulating protein expression levels of CyclinD1, p-AKT and p-CYLD and inducing tumor cell apoptosis. Animal model tests show that the compound remarkably inhibits the growth of mouse tumors and the tumor angiogenesis, has no obvious toxic or side effect, has better anti-tumor effect and broad application prospect, and can be used for developing an anti-tumor medicament.

Description

The antitumor application of 2-amino-4-(3 '-cyano group-4 '-pyrrolidinyl) phenylpyrimidine compound

Technical field

The invention belongs to tumor chemicals field, particularly relate to 2-amino-4-(3 '-cyano group-4 '-pyrrolidinyl) application of the antitumor of phenylpyrimidine compound.

Background technology

Chemotherapy be cancer clinical treatment important means, chemotherapeutics by directly kill tumor cell or inhibition tumor cell division growth, promote cell differentiation reach therapeutic effect.Seek active and effective prevention by way of and effective medicine seem very urgent and necessary, the particularly application of molecular targeted therapy in oncotherapy, find effective, special suppression means and inquire into its mechanism of action, not only be conducive to the generation of prophylaxis of cancer, new approach can also be provided for clinical treatment.

In recent years, people find other two kinds of non-classical I kappa b kinases (IKKs) gradually---TBK1 and IKKi, they are important derivants that body starts interferon signal path when being subject to viral infection, after Toll-like receptor is activated by virus composition, TBK1 and IKKi and TRAF3, TANK assemble, further at multiple serine and threonine residues site phosphorylation interferon regulatory factor (IRF 3,7), IRF-3 and IRF-7 dimerization, enter the expression of the multiple antiviral genes such as core induction interferon type Ⅰ gene, thus activate innate immunity approach.Except the role in antiviral immunity reaction, non-classical IKKs also participates in other signal paths, such as tumor generation etc.To it is reported in some transformant that TBK1 kinase activity raises and be that the existence of these transformants is necessary.Suppress the expression of TBK1 can the apoptosis of inducing tumor cell by RNAi technology, but the epithelial cell of non-tumorigenic but rely on TBK1 and survive.IKKi is the oncogene of human breast carcinoma to have genomics research to show, and in human breast cancer cell, suppress IKKi can inducing mammary cancer cell death.Thus TBK1 and IKKi causes people's attention gradually in oncology.

This research finds TBK1 and IKKi in same tumor and non-concurrent high expressed or intrinsic state of activation, suppresses one of them expression can activate another kinases.Therefore, due to the eclipsing effects between this two kinds of kinases and compensation, simple suppression TBK1 or IKKi all can not effectively grow by inhibition tumor cell, but this can effective inhibition tumor cell propagation ought to suppress these two kinds of kinases, has thus pointed out it may become the good target spot for the treatment of one of tumor simultaneously.

Summary of the invention

The present invention aims to provide a kind of 2-amino-4-(3 '-cyano group-4 '-pyrrolidinyl) anticancer usage of phenylpyrimidine compound, for new thinking and countermeasure expanded by the novel targeted suppression of Future Development TBK1 and IKKi antitumor drug.

In order to achieve the above object, technical scheme provided by the invention is:

The invention provides a kind of 2-amino-4-(3 '-cyano group-4 '-pyrrolidinyl with suppression TBK1 and IKKi) phenylpyrimidine compound, in this description, this compound is as follows referred to as CMPD, CMPD structural formula, and its association attributes is as shown in the table.

Structural formula of compound

2-amino-4-(3 '-cyano group-4 '-pyrrolidinyl) association attributes of phenyl pyrimidine

Present invention is disclosed Compound C MPD existence by suppressing the mechanism of TBK1 and IKKi inducing apoptosis of tumour cell, and found that CMPD is by suppressing the effect of TBK1 and IKKi anti-tumor in vivo.Discovery is studied through the present invention, between these two kinds of kinases of TBK1 and IKKi, there is eclipsing effects and compensation, simple suppression TBK1 or IKKi all can not effectively grow by inhibition tumor cell, but this can effectively breed by inhibition tumor cell ought to suppress these two kinds of kinases simultaneously, Compound C MPD then can suppress these two kinds of kinases simultaneously, thus becomes the therapy target of new type antineoplastic medicine.

The present invention also demonstrates the antitumor curative effect of Compound C MPD by animal model, CMPD successfully can suppress the growth of carcinoma of tongue tumor bearing nude mice and carcinoma of prostate FVB mouse interior tumor by TBK1 and IKKi double inhibition, and to mice body without obvious toxicity, side effect.The antitumor action of this compound activates with suppressing AKT, lowers vegf expression, Tumor suppression associated angiogenesis.The present invention is that the clinical medicine of development of new targeted inhibition TBK1 and IKKi further from now on lays the foundation.

Accompanying drawing explanation

Fig. 1 is that in the embodiment of the present invention 1, TBK1 and IKKi expresses and Activation in kinds of tumor cells.

Fig. 2 is the protein expression level of the IKKi of IKKi-siRNA SCC-25 cell and matched group in the embodiment of the present invention 1, p-TBK1, TBK1.

Fig. 3 is that in the embodiment of the present invention 1, in people's Human Tongue Carcinoma Lines SCC-25, siRNA suppresses TBK1 or/and its cell proliferation level after IKKi.(* * P<0.01 difference has statistical significance, and siRNA jamming effectiveness adopts Western blotting checking).

Fig. 4 is RAW264.7 cell LPS(1 μ g/ml in the embodiment of the present invention 2) process the protein expression level of IRF-3 and p-IRF-3 after different time.

Fig. 5 is after CMPD process 1 hr of RAW264.7 cell variable concentrations in the embodiment of the present invention 2 and with LPS(1 μ g/ml) stimulate the protein expression level of IRF-3 and p-IRF-3 of 1 hr.

Fig. 6 is CMPD(1.0 μM under inverted phase contrast microscope in the embodiment of the present invention 3) the cellular morphology change (a: contrast of the SCC-9 cell of process different time length; B:24 hr; C:48 hr; D:72 hr) (enlargement ratio 200 ×).

Fig. 7 be in the embodiment of the present invention 3 SCC-9 cell at CMPD(1.0 μM) process 72 hr after with untreated fish group comparison diagram (in figure, red-label is apoptotic cell, and blue markings is nucleus).

Fig. 8 be in the embodiment of the present invention 3 SCC-9 cell at CMPD(1.0 μM) related apoptosis protein expression level after different time process.

Fig. 9 is SCC-9 cell related apoptosis protein expression level after CMPD process 24 hr of variable concentrations in the embodiment of the present invention 3.

Figure 10 be in the embodiment of the present invention 3 SCC-9 cell at CMPD(1.0 μM) Cyclin D1 protein expression level after process different time process.

Figure 11 is that in the embodiment of the present invention 3, SCC-9 cell AKT and phosphorylated protein (S473, T308) thereof after CMPD process 30 min of variable concentrations is expressed.

Figure 12 be in the embodiment of the present invention 3 SCC-9 cell at CMPD(1.0 μM) process p-CYLD protein expression level after 1 hr.

Figure 13 be in the embodiment of the present invention 4 at the end of experiment in nude mice outward appearance photo and art tumor dissect photo.

Figure 14 is that the gross tumor volume of CMPD treatment carcinoma of tongue SCC-9 cell tumor bearing nude mice in the embodiment of the present invention 4 changes and tumor weight and tumor photo (* P<0.05 at the end of experiment; * P<0.01).

Figure 15 is that the gross tumor volume of CMPD treatment carcinoma of prostate Myc-CaP cell lotus tumor FVB mice in the embodiment of the present invention 4 changes and tumor weight and tumor photo (* P<0.05 at the end of experiment; * P<0.01).

Figure 16 is the expression (enlargement ratio 400 ×) of Immunohistochemical detection VEGF and p-AKT in matched group and experimental group nude mouse tumor tissue in the embodiment of the present invention 4.

Figure 17 is that in the embodiment of the present invention 4, nude mouse tumor organizes RT-PCR to detect VEGF gene expression dose (n=3).

Figure 18 is that in the embodiment of the present invention 4, nude mouse tumor organizes Western blotting to detect p-AKT protein expression level (n=3).

Detailed description of the invention

embodiment 1, TBK1 and the IKKi important function in tumor cell proliferation

Choose ten kinds of tumor cell lines, comprise longue carcinoma (SCC-9, SCC-15, SCC-25), Prostatic cancer cell lines (PC3, DU145, LNCaP, Myc-CaP), albumen is extracted after breast carcinoma cell strain (MCF-7, MDA-MB-231, T-47D) cellar culture, Western blotting measures the protein content of its TBK1, P-TBK1, IKKi, P-IKKi, and β-actin is internal reference (Fig. 1).

Found that TBK1 and IKKi all has expression in tumor cell, the phosphorylation level of IKKi is expressed all as seen in various tumor cell, but the phosphorylation level of TBK1 is expressed inconsistent in different tumor cell, even at longue carcinoma (SCC-9, SCC-15, SCC-25) in be difficult to detect that p-TBK1 expresses.But suppress its IKKi to express (Fig. 2) when adopting siRNA technology in SCC-25 cell, the expression of the TBK1 that Western blotting found that in the SCC-25 of IKKi-siRNA process is better than matched group SCC-25 cell, and the expression of its P-TBK1 simultaneously also obviously strengthens.Therefore illustrate that TBK1 and IKKi expresses and activate in tumor cell, the simple IKKi that suppresses can cause the compensatory activation of TBK1.

And then adopt siRNA to suppress TBK1 or/and IKKi can obtain con-siRNA to Human Tongue Carcinoma Lines SCC-25, TBK1-siRNA, IKKi-siRNA and TBK1 & IKKi-siRNA SCC-25 tetra-groups, cellar culture 5 days, respectively the the 0th, 1,3, within 5 days, carry out cell counting, be depicted as broken line graph (Fig. 3).TBK1-siRNA, the propagation level of IKKi-siRNA SCC-25 two groups of cells slightly declines relative to matched group, but they are difference not statistically significant (P>0.05) between three groups, but after TBK1 and IKKi of SCC-25 is all suppressed, its propagation level is significantly suppressed, and between its excess-three group, difference has statistical significance (P<0.01).This illustrates that siRNA technology suppresses separately the expression of TBK1 or IKKi effectively not suppress Human Tongue Carcinoma Lines to be bred, and suppresses to express both it, effectively can suppress the propagation of Human Tongue Carcinoma Lines when simultaneously.

Therefore embodiment shows, TBK1 and IKKi high expressed activating in tumor cell, suppresses them can the propagation of effective inhibition tumor cell simultaneously, shows that TBK1 and IKKi can become the molecular target of new type antineoplastic medicine.

embodiment 2, CMPD are to the rejection ability of TBK1 and IKKi and anti tumor activity in vitro evaluation experimental

TBK1 and IKKi mainly come from immunne response its activate after can phosphorylation IFN regulatory factor 3 and 7 (IRF-3/IRF-7), in RAW264.7 cell, LPS can activate TBK1 and IKKi phosphorylation IRF-3, and thus the suppression situation of TBK1 and IKKi activity can be detected by Western blotting in RAW264.7 cell.If these two kinds of kinase whose activity are blocked, the phosphorylation level of IRF-3 will reduce or disappear, and p-IRF-3 band just can embody compound indirectly to these two kinds of kinase whose rejection ability.

Fig. 4 is presented at LPS in this description of LPS(and refers to lipopolysaccharide) effect under, p-IRF-3 express strengthen gradually, the highest at 1 hr, reduce afterwards, be difficult to detect when 4 hr.Obtaining p-IRF-3 optimal detection condition is that LPS stimulates 1 hr.Thus to after CMPD process 1 hr of RAW264.7 cell variable concentrations and with LPS(1 μ g/ml) stimulate 1 hr, Western blotting detects the protein expression level of each group of IRF-3 and p-IRF-3 to analyze the rejection ability (Fig. 5) of itself TBK1 and IKKi, the phosphorylation level of IRF-3 all can be lowered under variable concentrations CMPD effect, the phosphorylation level of IRF-3 just effectively can be reduced under 0.5 μM of effect, and 1.0 μMs time, p-IRF-3 almost cannot be detected, therefore can think that this compound has the rejection ability of TBK1 and IKKi relatively preferably.

In various tumor cell line, use the CPMD of gradient concentration to act on 72 hr respectively, in isocyatic this description of DMSO(, DMSO refers to dimethyl sulfoxide) be set to contrast, mtt assay detects proliferative activity.IC50 value (IC50 value refers to the drug concentration of energy antiproliferative effect 50%) can be calculated according to above MTT result.Generally speaking, IC50 is lower, and the antitumor action of compound is stronger.Compound C MPD has good Inhibit proliferaton effect at selected tumor cell line, shows as its IC50 lower, all at micromole's order of magnitude (0.584-1.872 μM), as shown in table 1, shows higher anti-tumour cell proliferative activity.

table 1compound C MPD is to the IC50 of various Proliferation of Tumor Cells In Vitro

Tumor cell line	IC50 (μM)
		SCC-9	0.800
SCC-15	0.645
		SCC-25	0.678
PC3	0.667
		DU145	0.584
LNCaP	0.584
		Myc-CaP	0.601
MCF-7	0.899
		MDA-MB-231	1.872
T-47D	0.876

Therefore embodiment 2 shows, this TBK1 and IKKi of CMPD double inhibition compound has good pharmacological properties, and can be used as the candidate compound of new type antineoplastic medicine, it all has the effect of obvious Inhibit proliferaton to selected tumor cell.

embodiment 3, CMPD promote apoptosis of tumor cells and Mechanism Study

To SCC-9 cell CMPD(1.0 μM) process, Microscopic observation is carried out and take pictures (Fig. 6) to 72hr after 48 hr after 24 hr, process before process, after process and process, SCC-9 cell attachment growth before treatment, cellular morphology is polygon or oval, be evenly distributed, cytochrome oxidase isozymes is connected to each other.After acting on 24 hr, part Cell tracking ruptures, and cellular morphology systematicness reduces, and has fusiformis with similar not too regular triangle; After acting on 48 hr, occur wide crack between attached cell, part cell becomes circle, volume-diminished, and soft edge is unintelligible, and part cell membrane is little blister bulging, and part cell detachment suspends; After acting on 72 hr, it is more that cell detachment suspends, and attached cell is rare, and cell is irregular, not of uniform size, is even cracked into fragment.This phenomenon shows, along with extended durations of action, CMPD is induction of the apoptosis of Human Tongue Carcinoma Lines SCC-9.

Further use ApopTag original position apoptosis detection kit 72 hr are cultivated to SCC-9 cell routine and with Compound C MPD(1.0 μM) these two groups of cells of SCC-9 cell process 72 hr are detected (Fig. 7).Every picture group sheet has three figure, and in first (DAPI), blue portion is positioning cells core, i.e. all cells; In second (ApoTag) there is the cell of apoptosis in RED sector location; 3rd (Merge) is by first and the second opening and closing and forms, and can observe the cell of all cells and generation apoptosis simultaneously.Therefore, in figure, the ratio of matched group apoptotic cell is starkly lower than CMPD processed group; On the other hand, the photo that simple observation DAPI dyes can find that the blue portion of CMPD processed group presents ellipse unlike matched group, but irregular, lamellar distribution, after this means CMPD process there is fragmentation in nuclear DNA, blue portion quantity is also few compared with matched group, and after which illustrating CMPD process, cell proliferation rate is also less than matched group.This also demonstrates the result of morphological observation and last chapter ganglion cell proliferation experiment gained above, again illustrates that CMPD can promote Human Tongue Carcinoma Lines apoptosis, inhibits it to breed.

Use CMPD(1.0 μM respectively) gradient timetable treatment S CC-9 cell (Fig. 8), and use the CMPD of gradient concentration to SCC-9 cell process 24 hr(Fig. 9), Western blotting detects relevant Caspase family protein and PARP Activation.Result is presented at the growth along with the processing time under same CMPD concentration, and Cleaved Caspase-9, Cleaved Caspase-3, Cleaved Caspase-7 and Cleaved PARP protein expression level strengthen gradually; Increasing along with CMPD concentration of same processing time, its apoptosis-related protein expression is same variation tendency.Illustrate that CMPD induces Human Tongue Carcinoma Lines apoptosis, in the time with dose dependent.

With CMPD(1.0 μM) gradient timetable process is carried out to carcinoma of tongue SCC-9 cell, Western blotting detects the expression (Figure 10) of Cyclin D1 albumen, and result display is along with increasing the action time of CMPD, and Cyclin D1 expresses decline.

With CMPD effect SCC-9 30 min of gradient concentration, Western blotting detects the expression (Figure 11) of AKT and phosphorylated protein thereof, result shows along with CMPD concentration increases, the phosphorylation level in two sites of AKT declines, in dose dependent, show that this TBK1 and IKKi double inhibition compound can suppress the activation of AKT.

With CMPD(1.0 μM) process carcinoma of tongue SCC-9 cell 1 hr, Western blotting detects the expression (Figure 12) of p-CYLD albumen, and after result display CMPD process, its p-CYLD expresses and all has reduction.

Therefore this example confirms, CMPD has double inhibition TBK1 and IKKi function, can induce Human Tongue Carcinoma Lines apoptosis, and presentative time and dose dependent, its mechanism is relevant with the protein expression lowering Human Tongue Carcinoma Lines Cyclin D1, p-AKT, p-CYLD.

embodiment 4, CMPD study at the experiment in vivo of animal model for tumour

Based on Compound C MPD, there is double inhibition TBK1 and IKKi extracorporeal anti-tumor ability of cell proliferation, adopt bearing mouse model to verify the ability that its antitumor is bred.

Tumor cell line adopts Human Tongue Carcinoma Lines SCC-9, be purchased from American Type Culture collection warehousing (American type culture collection, and prostate cancer cell line Myc-CaP ATCC), separate from a kind of c-Myc transgenic carcinoma of prostate mice, it can grow tumor in the immune FVB Mice Body of tool.

BALB/C innate cells immunodeficiency Nu/Nu nude mouse 15 is selected in carcinoma of tongue zoopery, female, 8 week age, body weight 22-24 g, SPF(Special pathogen free) level.Myc-CaP Prostate Cancer Animal experiment employing 12 FVB mices.At aseptic independent air-feeding isolation cage tool lamina air flow purification indoor feeding after all animals arrive, keep constant temperature (26-28 DEG C), constant humidity (relative humidity 40%-60%), 12 hr light and shades switch.Feedstuff, drinking-water, bedding and padding all use after sterilization treatment.Experiment, feeding process all reach SPF condition.

(1) carcinoma of tongue SCC-9 cell tumor bearing nude mice is treated with CMPD.

(1) preparation cell is collected

Choose exponential phase SCC-9 cell, digestion, centrifugal collecting cell, PBS washs 2 times, counted under microscope, and adds appropriate Matrigel, and adjustment final concentration of cells is 2.0 × 107/ml cell suspension (operates on ice, keep low temperature) for subsequent use.

(2) animal is inoculated

After nude mice arrives at reception, in order to allow nude mice adapt to new environment, regulate state, first about animal center breeding observing l week, the life of record nude mice and health.Wait for nude mice diet, movable normal and test after levying without excitation.In superclean bench, neck skin of back on the left of routine disinfection nude mice, accurately extracts cell suspension 0.5 ml(namely containing 1 × 107, SCC-9 cell with l ml asepsis injector) inject neck dorsal sc on the left of nude mice, point of puncture is apart from injection site about 1 cm.Send nude mice back to former rearging cage after inoculation, observe and record the diet of nude mice, spirit and defecation situation.

(3) grouping and treatment

After inoculating 24 hr, 15 nude mices are divided into 2 groups at random, matched group 5, experimental group 10.Treatment group every day (same time point) is to nude mice abdominal cavity injection CMPD solution, dosage is that (nude mice as 20 mg needs 2 mg compounds to 100 mg/kg body weight, namely 200 ul Compound C MPD solution (1 mg/100 ul) are injected), continuous treatment 34 days, matched group (time point same with experimental group) injecting normal saline is (the nude mice direct injection 200 ul normal saline of 20 mg) in contrast, and two groups of nude mices terminate treatment simultaneously.

(2) carcinoma of prostate Myc-CaP cell lotus tumor FVB mice is treated with CMPD

(1) preparation cell is collected

Choose exponential phase Myc-CaP cell, digestion, centrifugal collecting cell, PBS washs 2 times, counted under microscope, and adds appropriate Matrigel, and adjustment final concentration of cells is 6.0 × 106/ml cell suspension (operates on ice, keep low temperature) for subsequent use.

(2) animal is inoculated

Rearing conditions is as above-mentioned, in superclean bench, neck skin of back on the left of routine disinfection FVB, accurately extracts cell suspension 0.5 ml(namely containing 3.0 × 106, Myc-CaP cell with l ml asepsis injector) inject neck dorsal sc on the left of mice, point of puncture is apart from injection site about 1 cm.Send mice back to former rearging cage after inoculation, observe and record the diet of mice, spirit and defecation situation.

(3) grouping and treatment

After inoculating 24 hr, 12 FVB mices are divided into 2 groups at random, experimental group 6, matched group 6.Treatment group every day (same time point) is to mouse peritoneal injection CMPD solution, dosage is 100 mg/kg body weight, continuous treatment 18 days, matched group (time point same with experimental group) injecting normal saline is (the nude mice direct injection 200 ul normal saline of 20 mg) in contrast, and two groups of FVB mices terminate treatment simultaneously.

Curative effect evaluation

Conventional to raise, observe the diet of mice, activity every day, within every three days, survey Mouse Weight, and with the major diameter (L) of vernier caliper measurement tumor, minor axis (W), and record.Its gross tumor volume computational methods are shown in formula below: square (W2)/2 of tumor volume (V)=tumor major diameter (L) × tumor minor axis.

Results of animal

Two groups of tumor cell inoculations are after mice, and small embossment appears in its injection site, are tumor tuberosity, in not too regular circle, and some uneven surface.Matched group tumor is very fast in growth, and centered by initial injection position, infiltrate towards periphery gradually, volume increases gradually, and the quality touched is harder.Minority tumor center occurs downright bad, depression or incrustation phenomenon; Experimental group tumor growth is slow, and volume is less than matched group.Along with the compound lumbar injection time increases in experimentation, it is obvious gradually to the inhibition of experimental group tumor growth.

Put to death whole mice at the end of experiment, anatomical isolation goes out tumor, and have more blood vessel to grow into around visible matched group tumor in art, tumor body colour pool is ruddy; And experimental group tumor peripheral vessels is tiny, sparse, tumor body color partially light (Figure 13).Figure 14 and Figure 15 be shown in by the picture of whole tumor, and matched group tumor is obviously greater than experimental group, and in illustrative experiment group, the growth of tumor is suppressed under the intervention of CMPD.

CMPD toxicity assessment: experiment start and at the end of participate in test Mouse Weight be showed no notable difference.In therapeutic process, the mice mental status is good, and growth promoter, diet activity are showed no obvious abnormalities, and shows that CMPD is without obvious toxicity, side effect.

Frozen section is carried out to fresh nude mouse tumor tissue, and carries out immunohistochemistry, gather image and see Figure 16.In this description of VEGF(, VEGF refers to VEGF) mainly to express in endochylema, positive particle is brown color, is wherein strongly expressed in matched group, in weak expression in experimental group.P-AKT then all has expression in nucleus and endochylema, wherein in matched group in nucleus and endochylema all in strongly expressed, in experimental group in nucleus and endochylema in weak expression (Figure 16).Indicate CMPD treatment nude mice carcinoma of tongue and can lower itself VEGF and p-AKT.

RT-PCR detects the gene expression dose (Figure 17) of VEGF in nude mouse tumor tissue, and the gene expression dose of result display VEGF is obviously lowered in the experimental group of CMPD process, and this result is consistent with immunohistochemical analysis result above.

Western blotting detects the protein expression level (Figure 18) that nude mouse tumor organizes p-AKT, the protein expression level of result display p-AKT is obviously lowered in the experimental group of CMPD process, and Western blotting analysis result is consistent with immunohistochemical analysis result above.

Therefore this example confirms, this TBK1 and IKKi double inhibition of CMPD compound successfully can suppress the growth of carcinoma of tongue tumor bearing nude mice and carcinoma of prostate FVB mouse interior tumor, and to mice body without obvious toxicity, side effect.The antitumor action of CMPD activates with suppressing AKT, lowers vegf expression, Tumor suppression associated angiogenesis.

Claims (2)

1.2-amino-4-(3 '-cyano group-4 '-pyrrolidinyl) application of phenylpyrimidine compound in the anti-tongue cancer drug of preparation, the structural formula of described compound is:

。

2.2-amino-4-(3 '-cyano group-4 '-pyrrolidinyl) phenylpyrimidine compound preparing the application in antiprostate cancer, and the structural formula of described compound is:

。