CN106188028A - Containing oxadiazoles heterocycle compound and its preparation method and application - Google Patents

Abstract

The invention provides containing oxadiazoles heterocycle compound and its preparation method and application, wherein, the heterocycle compound containing oxadiazoles has a following general structure:Found by cellular level experiment, the propagation that all can suppress K562-3d, HL60, KG1a cell containing oxadiazoles heterocycle compound that the present invention provides, particularly example 11 compound (compound 10c) is in the case of drug level is less than positive control medicine imatinib, can realize suppressing the effect of K562-3d, HL60, KG1a cell proliferation, the heterocycle compound containing oxadiazoles that therefore present invention provides to may be used for treating leukemia equally.

Description

Containing oxadiazoles heterocycle compound and its preparation method and application

Technical field

The present invention relates to containing oxadiazoles heterocycle compound and its preparation method and application.

Background technology

Leukemia, also referred to as leukemia, be the Clonal malignant disease of a class hematopoietic stem cell exception.ItsLeukaemia in clone loses and is further differentiated into ripe ability and is stuck in cytocerastic differenceStage.In bone marrow and other hemopoietic tissue, a large amount of hypertrophy of leukaemia is gathered and infiltrates other organsAnd tissue, make normal hematopoiesis suppressed simultaneously, clinical manifestation is anemia, hemorrhage, infection and each organInfiltration symptom.

Leukemia is generally divided into: 1) acute leukemia；2) chronic leukemia；3) the white blood of specific typeSick.

Acute leukemia is the Clonal malignant disease that a kind of hematopoietic stem cell is abnormal.Its clone in whiteDisorders of blood cell loses and is further differentiated into ripe ability and is stuck in cytocerastic different phase.At boneIn marrow and other hemopoietic tissue, a large amount of hypertrophy of leukaemia is gathered and infiltrates other organs and tissue, withTime make normal hematopoiesis suppressed, clinical manifestation be anemia, hemorrhage, infect and each organ infiltration's symptom.Acute leukemia generally can be divided into Lymphocytic leukemia or granulocyte leukemia, and they can be fromDifferentiate in phenotype and gene, be characterized in that course advancement is quick, need to treat immediately.Acute leukemiaDerive from hemopoietic progenitor cell in early days.Acute lymphoblastic leukemia is the malignant tumor that comparison is rare.OftenThe total incidence in year acute lymphoblastic leukemia (ALL) is 1.1/100,000.This sickness rate is in childhoodPeriod peaks, and is gradually increased along with the age and constantly reduces.After 35 years old, this sickness rate is the most againSecondary rising, and second peak it was initially observed from 80 years old.

The treatment of acute lymphoblastic leukemia (ALL) adult patient becomes day by day complicated, because forDifferent disease subtypes introduces diversified therapeutic scheme, embodies according to being adapted to particular riskSick plant the intention most preferably adjusting treatment.Complete up-to-date improvement by the new therapeutic scheme of introducing,As added tyrosine kinase inhibitor imatinib or pin in early days for positive (Ph+) ALL of Ph-The CD20+ case of B-system ALL is used anti-CD 20 antibodies Rituximab (Rituximab).ButIt is these medicines curative effect extreme differences to Recurrent Acute Lymphocytic leukemia (ALL) patient, about 30%Patient will finally be recurred.

Characteristic type leukemia can include again promyelocytic leukemia, Plasmacytic leukemia, lymphomaLeukemia and central leukemia.This specific type leukemia, faces except having general leukemiaBed performance (heating, hemorrhage, anemia), infiltration symptom, liver splenic lymph nodes enlargement) outward, mainly haveFollowing features: morbidity is anxious, progress is fast, hemorrhage substantially, mortality cerebral hemorrhage can occur or merge disperseProperty intravascular coagulation, as failed to diagnose in time and treat, can cause death because of hemorrhage at short notice.Therefore, it is the stealthy killer of human health.

Wherein, chronic myelocytic leukemia (chronic myelocytic leukemia, CML) is named againChronic myelocytic leukemia, it is the hematopoietic stem cell malignant clone proliferative disease of blood system, leadsThere is the symptom such as anemia, hemorrhage, infection in pathogenic people.CML accounts for about the 20% of adult leukemia, greatlyThe most annual sickness rate is 50,000/, it is likely to morbidity at any age bracket, especially existsIn old people, sickness rate is higher, and male is higher than women sickness rate.Progression of disease according to CML patientSituation and the order of severity are divided into chronic phase (CP), accelerated period (AP) and acute transformation phase (BP).AboutThe patients with chronic myelocytic leukemia of 85%～90% is diagnosed discovery in chronic phase, and this stage symptom is alsoInconspicuous or the most weak, arthralgia or abdominal distention, if Drug therapy is timely, chronic phase, is justCan be elongated, in the case of not treatment in time, disease can enter into accelerated period.The master that CML is formedWanting reason is No. 9 and No. 22 chromosomes generation dystopy t (9:22) (q34:q11) (Philadelphia chromosome Ph)Forming a kind of fusion gene i.e. BCR-ABL fusion gene, this gene can encode BCR-ABL protein,It it is a kind of active tyrosine kinase.This BCR-ABL kinases can make some substrate phosphorylations in downstream,And then activate the signal transduction pathway relevant with cell proliferation, cause ripe granulocyte infinite multiplication to leadCause the generation of CML.

At present, Therapeutic Method to CML is mainly traditional Drug therapy, hematopoietic stem cell clinicallyTransplant and emerging molecular targeted therapy.Common Drug therapy include alkylating agent busulfan, hydroxyurea,Interferon (IFN-α), cytosine arabinoside and homoharringtonine.Wherein, due to busulfan side reaction ratioRelatively big, stop using；Although hydroxyurea is cheap, treatment cost is low,Toxic and side effects is little, but it is poor for the patient's CML action effect being in accelerated period, and for urgencyThe CML patient of change phase does not has effect substantially；Alpha-interferon is less compared with the above two toxic and side effects, Bing RennaiMight as well by property, it is possible to produce lasting cytogenetics and alleviate the life span extension making CML patient,But it is very poor to the CML patient outcome of accelerated period and acute transformation phase.One of this common drug treatmentThe biggest drawback is also to produce normal body cell while tumor cell is produced lethal effectDamage.Hematopoietic stem cell transplantation is the method that the most uniquely can cure CML at present, and it is to pass throughFirst with Large dose chemoradiotherapy, CML tumor cell in the patient is removed, then making autologous or allosomeHemocytoblast is transplanted to patient so that it is re-establish normal hematolymphiod system.But this sideMethod gets up to there is a lot of difficulty in practical operation, and there is certain requirement at the such as age to patient, operationComplexity, success rate is low, expends height, and it is necessary to have suitable Marrow donation person, and transplant the later stageIt is likely to occur infection, CML patient and household can be made to face the biggest risk, so hematopoietic stem cellIt is implanted in actual application the most restricted.Until first, the calendar year 2001 whole world occurs relevant to tumorThe listing of signal transduction inhibitor imatinib, the hope making the treatment of CML see, start CMLThe New Times of molecular targeted therapy.

Imatinib is that first, the whole world is that get the Green Light there is relevant signal conduction suppression to tumorAgent, is mainly used in treating acute and chronic leukemia, in recent years, along with imatinib is slow as treatmentOne line medication of property granulocyte leukemia is widely used, and its drug resistance phenomenon the most gradually highlights, and manyOccur CML accelerated period or the patient of acute transformation phase.So finding one leukemic stem cells is had killingPower and the compound of imatinib-resistant can be overcome extremely urgent, and unlatching CML is also controlled by thisThe new direction treated and the clinical assessment standard changing anti-CML medicine.

Above-mentioned data show, leukemia remains explosive for Most patients and cannot cureDisease.Thus, in the urgent need to the leukemic medicine for the treatment of improved.

Summary of the invention

In order to solve for treating the problem that leukemic curative effect of medication is the best in prior art, according to thisOne aspect of invention, it is provided that a kind of heterocycle compound Han oxadiazoles, described containing oxadiazoles heterocycleCompounds has a following general structure:

Above-mentioned containing in oxadiazoles heterocycle compound, in described formula, R includes following group:

According to a further aspect in the invention, a kind of preparation containing oxadiazoles heterocycle compound is additionally providedMethod, it is characterised in that include following synthetic route: make N-acyl group-4-methyl-3-((4-(pyridine-3-)Pyrimidine-2-) amino) benzoyl hydrazine compound generation rearrangement reaction, it is converted into the described heterocyclic Han oxadiazolesCompound；Wherein, described N-acyl group-4-methyl-3-((4-(pyridine-3-) pyrimidine-2-) amino) benzoyl hydrazineCompound has a following general structure:

And

The described heterocycle compound containing oxadiazoles has a following general structure:

In above-mentioned preparation method, described N-acyl group-4-methyl-3-((4-(pyridine-3-) pyrimidine-2-) amino)In the general structure of benzoyl hydrazine compound and the described general structure containing oxadiazoles heterocycle compound, RIncluding following group:

In above-mentioned preparation method, under conditions of Trifluoromethanesulfonic anhydride and triphenylphosphine oxide exist orUnder conditions of potassium carbonate and paratoluensulfonyl chloride exist, there is described rearrangement reaction.

In above-mentioned preparation method, described N-acyl group-4-methyl-3-((4-(pyridine-3-) pyrimidine-2-) amino)The preparation method of benzoyl hydrazine compound farther includes: at 1-(3-dimethylamino-propyl)-3-ethyl carbonDiimmonium salt hydrochlorate (EDCl), I-hydroxybenzotriazole (HOBT) and N-methylmorpholine (NMM)In the presence of, make 4-methyl-3-{ [4-(pyridine radicals-3-base) pyrimidine radicals-2-base] amino } benzoyl hydrazine and carboxylic acid chemical combinationThing (R-CH3COOH) reacts, and generates described N-acyl group-4-methyl-3-((4-(pyridine-3-) pyrimidine-2-)Amino) benzoyl hydrazine compound；

Wherein, R includes following group:

According to another aspect of the invention, additionally provide above-mentioned containing oxadiazoles heterocycle compound for controllingTreat the application in leukemic medicine.

In above-mentioned application, described for treating leukemic pharmaceutical pack containing described containing oxadiazoles heterocyclicCompound, the most acceptable salt of the described heterocycle compound containing oxadiazoles, ester, hydrate orCombinations thereof and adjuvant.

In above-mentioned application, the dosage form of the leukemic medicine of described treatment is selected from tablet, capsule, ballAgent, suppository, aerosol, oral liquid, granule, powder, injection, syrup, wineAgent, tincture, distillate medicinal water, membrane or combinations thereof.

In above-mentioned application, the administering mode of the leukemic medicine of described treatment includes being administered orally, injects,Implantation, external, spray, suck or combinations thereof.

Thus, the invention provides a class new containing oxadiazoles heterocycle compound and preparation method thereof and shouldWith, found by cellular level experiment, what the present invention provided all can press down containing oxadiazoles heterocycle compoundThe propagation of K562-3d, HL60, KG1a cell processed, by measuring IC₅₀And IC₉₀Value finds, the present inventionThere is provided makees the suppression of the propagation of K562-3d, HL60, KG1a cell containing oxadiazoles heterocycle compoundWith all can reach 90%, and example 11 compound (compound 10c) that particularly present invention provides existsDrug level less than in the case of positive control medicine imatinib, can realize equally suppressing K562-3d,The effect of HL60, KG1a cell proliferation, what therefore the present invention provided can containing oxadiazoles heterocycle compoundFor treatment leukemia.

Accompanying drawing explanation

Fig. 1 is compound 6a¹H NMR。

Fig. 2 is compound 6a¹³C NMR。

Fig. 3 is the HR-MS of compound 6a.

Fig. 4 is compound 6b¹H NMR。

Fig. 5 is compound 6b¹³C NMR。

Fig. 6 is the HR-MS of compound 6b.

Fig. 7 is compound 6c¹H NMR。

Fig. 8 is compound 6c¹³C NMR。

Fig. 9 is the HR-MS of compound 6c.

Figure 10 is compound 6d¹H NMR。

Figure 11 is compound 6d¹³C NMR

Figure 12 is the HR-MS of compound 6d.

Figure 13 is compound 6e¹H NMR。

Figure 14 is compound 6e¹³C NMR。

Figure 15 is the HR-MS of compound 6e.

Figure 16 is compound 6f¹H NMR。

Figure 17 is compound 6f¹³C NMR。

Figure 18 is the HR-MS of compound 6f.

Figure 19 is compound 6g¹H NMR。

Figure 20 is compound 6g¹³C NMR。

Figure 21 is the HR-MS of compound 6g.

Figure 22 is compound 6h¹H NMR。

Figure 23 is compound 6h¹³C NMR。

Figure 24 is the HR-MS of compound 6h.

Figure 25 is compound 10a¹H NMR。

Figure 26 is compound 10a¹³C NMR。

Figure 27 is the HR-MS of compound 10a.

Figure 28 is compound 10b¹H NMR。

Figure 29 is compound 10b¹³C NMR。

Figure 30 is the HR-MS of compound 10b.

Figure 31 is compound 10c¹H NMR。

Figure 32 is compound 10c¹³C NMR。

Figure 33 is the HR-MS of compound 10c.

Figure 34 is compound 10d¹H NMR。

Figure 35 is compound 10d¹³C NMR。

Figure 36 is the HR-MS of compound 10d.

Figure 37 is compound 10e¹H NMR。

Figure 38 is compound 10e¹³C NMR。

Figure 39 is the HR-MS of compound 10e.

Figure 40 is compound 10f¹H NMR。

Figure 41 is compound 10f¹³C NMR。

Figure 42 is the HR-MS of compound 10f.

Figure 43 is compound 10g¹H NMR。

Figure 44 is compound 10g¹³C NMR。

Figure 45 is the HR-MS of compound 10g.

Figure 46 is compound 10h¹H NMR。

Figure 47 is compound 10h¹³C NMR。

Figure 48 is the HR-MS of compound 10h.

Figure 49 is compound 10i¹H NMR。

Figure 50 is compound 10i¹³C NMR。

Figure 51 is the HR-MS of compound 10i.

Figure 52 shows imatinib, the suppression to K562 cell of the example 1-9 compound.

Figure 53 shows the suppression to K562 cell of the example 10-17 compound.

Figure 54 shows imatinib, the suppression to HL60 of the example 1-9 compound.

Figure 55 shows the suppression to HL60 of the example 10-17 compound.

Figure 56 shows imatinib, the suppression to KG1a cell of the example 1-9 compound.

Figure 57 shows the suppression to KG1a cell of the example 10-17 compound.

Detailed description of the invention

Below in conjunction with the accompanying drawing in the embodiment of the present invention, the technical scheme in the embodiment of the present invention is enteredRow clearly and completely describes, it is clear that described embodiment is only a part of embodiment of the present invention,Rather than whole embodiments.Based on the embodiment in the present invention, those of ordinary skill in the art are obtainedThe every other embodiment obtained, broadly falls into the scope of protection of the invention.

The chemosynthesis containing oxadiazoles heterocycle compound of embodiment 1 present invention offer and Structural Identification

1. the heterocycle compound containing oxadiazoles that the present invention provides has a following general structure:

In said structure formula, substituent group-R includes following group:

2. the experiment of synthetic chemistry step containing oxadiazoles heterocycle compound that the present invention provides:

2.1 general synthetic routes are:

2.2.1 the synthesis of compound 5a-5h

2.2.1.1 the conjunction of compound 2:4-methyl-3-[4-(pyridine radicals-3-base) pyrimidine radicals-2-base] Methyl anthranilateBecome

The absolute methanol of 30mL, compound 1 (1g, 3.26mmol), Xiang Qi is added in there-necked flaskIn be slowly added dropwise the concentrated sulphuric acid of 2mL, after dropping, reactant liquor is heated to reflux 3 hours at 65 DEG C,Rotation, except methanol, adds saturated aqueous sodium carbonate, filters, and filter cake is washed, and is dried to obtain yellow-brown solid 955Mg, productivity: 91.32%.

MS(ESI⁺): 321.6

¹H NMR(400MHz,CDCl₃): δ ppm 9.32 (s, 1H), 9.01 (s, 1H), 8.74 (d, J=3.6Hz, 1H), 8.53 (d, J=5.2Hz, 1H), 8.45 (d, J=8.0Hz, 1H), 7.73 (dd, J=1.2,7.6Hz, 1H), 7.45 (dd, J=4.8,8.0Hz, 1H), 7.30 (d, J=8.0Hz, 1H), 7.23 (d, J=5.2Hz,1H),7.15(s,1H),3.97(s,3H),2.42(s,3H)。

2.2.1.2 compound 3:4-methyl-3-{ [4-(pyridine radicals-3-base) pyrimidine radicals-2-base] amino } conjunction of benzoyl hydrazineBecome

In there-necked flask, add compound 2 (700mg, 2.185mmol), 10mL ethanol and 30mLHydrazine hydrate, is heated to 78 DEG C and refluxes 3 hours, cooling, and sucking filtration is washed twice, and cold ethanol washes 2Secondary, it is thus achieved that milk yellow solid 650mg, productivity: 92.86%.

MS(ESI⁺): 321.1

¹H NMR(400MHz,DMSO-d₆): δ ppm9.69 (s, 1H), 9.26 (d, J=1.6Hz, 1H),9.05 (s, 1H), 8.69-8.70 (m, 1H), 8.53 (d, J=4.8Hz, 1H), 8.44 (d, J=8.0Hz, 1H), 8.10 (s, 1H), 7.52-7.56 (m, 2H), 7.46 (d, J=5.2Hz, 1H), 7.31 (d, J=7.6Hz,1H),2.29(s,3H)

2.2.1.3 compound 5a-5h synthesizes

Compound 5a:4-methyl-N'-[3-(4-methyl isophthalic acid H-imidazole radicals-1-base)-5-(trifluoromethyl) benzoylBase]-3-{ [4-(pyridine radicals-3-base) pyrimidine radicals-2-base] amino } synthesis of benzoyl hydrazine

At room temperature, there-necked flask adds compound 4a:3-(4-methyl-1 H-imidazole-1-group)-5-(fluoroformBase) benzoic acid (50mg, 0.185mmol), EDCI (42.6mg, 0.222mmol), HOBT H₂O(34mg, 0.222mmol), NMM (30.5uL, 0.277mmol) and dichloromethane 5mL, stirsAfter mixing 20 minutes, add compound 3 (59.3mg, 0.185mmol), 24h is stirred at room temperature, with saturatedSodium carbonate regulation pH extracts to alkalescence, dichloromethane, and dichloromethane layer anhydrous sodium sulfate is dried, and filters,Filtrate rotation, except solvent, residue column chromatography purification, uses CH₂Cl₂: MeOH=30:1 to CH₂Cl₂:MeOH=15:1 eluting, it is thus achieved that white solid 67.2mg, productivity: 63.43%.

MS(ESI^-): 571.05

¹H NMR(400MHz,DMSO-d₆): δ ppm 10.71 (s, 2H), 9.27 (s, 1H), 9.14 (s, 1H), 8.70 (d, J=4.8Hz, 1H), 8.55 (d, J=5.2Hz, 1H), 8.45 (d, J=8.0Hz, 1H),8.42(s,1H),8.39(s,1H),8.28(s,1H),8.23(s,1H),8.16(s,1H),7.70(s,1H),7.69 (s, 1H), 7.55 (dd, J=5.2,8.0Hz, 1H), 7.47 (d, J=5.2Hz, 1H), 7.41 (d, J=4Hz,1H),2.35(s,3H),2.19(s,3H)

5b-5h has been obtained according to above-mentioned route and method.

The fluoro-N'-of compound 5b:4-(4-methyl-3-{ [4-(pyridine radicals-3-base) pyrimidine radicals-2-base] amino } phenyl)-3-(threeMethyl fluoride) benzoyl hydrazine

White solid 700mg, productivity: 87.9%.

¹H NMR(400MHz,DMSO-d₆): δ 10.68 (s, 2H), 9.27 (d, J=1.2Hz, 1H),9.13 (s, 1H), 8.70 (dd, J=1.6,4.8Hz, 1H), 8.54 (d, J=4.82Hz, 1H), 8.44-8.66(m, 1H), 8.31-8.35 (m, 2H), 8.21 (s, 1H), 7.66-7.74 (m, 2H), 7.55 (dd, J=4.8,7.2Hz, 3H), 7.47 (d, J=5.2Hz, 1H), 7.40 (s, J=8.0Hz, 1H), 2.34 (s, 3H).

Compound 5c:N'-[3,5-bis-(trifluoromethyl) phenyl]-4-methyl-3-{ [4-(pyridine radicals-3-base) pyrimidine radicals-2-base]Amino } benzoyl hydrazine

White solid, productivity: 73.7%.

¹H NMR(400MHz,DMSO-d₆): δ ppm 11.05 (m, 1H), 10.66 (s, 1H), 9.27 (d, J=1.6Hz, 1H), 9.14 (s, 1H), 8.70 (d, J=1.2,4.4Hz, 1H), 8.58 (s, 2H), 8.55 (d, J=5.2Hz,1H),8.43-8.46(m,2H),8.12(s,1H),8.15(s,1H),7.68(dd,1H),8.16(s,1H), 7.70 (s, J=1.6,8.0Hz, 1H), 7.54 (dd, J=4.8,8.0Hz, 1H), 7.48 (d, J=5.2Hz,1H), 7.42 (d, J=8.0Hz, 1H), 2.35 (s, 3H).

Compound 5d:4-methyl-N'-(4-methyl-3-{ [4-(pyridine radicals-3-base) pyrimidine radicals-2-base] amino } benzeneBase)-3-(trifluoromethyl) benzoyl hydrazine

Obtain white solid, productivity: 56%.

¹H NMR(400MHz,DMSO-d₆): δ ppm 9.28 (s, 1H), 8.66 (d, J=4.4Hz, 1H),8.58 (d, J=7.6Hz, 1H), 8.51 (d, J=5.2Hz, 1H), 8.45 (s, 1H), 8.26 (s, 1H), 8.08(d, J=8.0Hz, 1H), 7.69 (d, J=7.6Hz, 1H), 7.55-7.62 (m, 2H), 7.41-7.44 (m, 2H),2.57(s,3H),2.43(s,3H)。

Compound 5e:4-methyl-N'-[3-(2-methyl isophthalic acid H-imidazole radicals-1-base)-5-(trifluoromethyl) benzoylBase]-3-{ [4-(pyridine radicals-3-base) pyrimidine radicals-2-base] amino } benzoyl hydrazine

White solid, productivity: 56%.

¹H NMR(400MHz,DMSO-d₆): δ ppm 10.91 (s, 1H), 10.65 (s, 1H), 9.27 (d, J=1.6Hz, 1H), 9.14 (s, 1H), 8.69-8.70 (m, 1H), 8.55 (d, J=5.2Hz, 1H), 8.44 (d, J=8.0Hz, 1H), 8.34 (s, 1H), 8.26 (s, 1H), 8.24 (s, 1H), 8.17 (s, 1H), 7.69 (d, J=8.0Hz, 1H), 7.69 (s, 1H), 7.53-7.56 (m, 1H), 7.50 (s, 1H), 7.47 (d, J=5.2Hz, 1H),7.41 (d, J=8.0Hz, 1H), 6.99 (s, 1H), 2.37 (s, 3H), 2.35 (s, 3H).

Compound 5f:4-methyl-N-(4-((4-methyl piperazine-1-) methyl) benzoyl)-3-((4-(pyridine-3-) pyrimidine-2-)Amino) benzoyl hydrazine

White solid, productivity 84.5%.ESI-MS:[M+H]⁺537.3；¹HNMR(400MHz,MeOD-d₄): δ ppm 9.25 (s, 1H), 8.63 (d, J=4.0Hz, 1H), 8.55 (d, J=8.0Hz), 8.48(d, J=5.2Hz, 1H), 8.43 (s, 1H), 7.91 (d, J=8.0Hz, 2H), 7.67 (d, J=8.0Hz, 1H),7.57 (dd, J=5.2Hz, 8.0Hz, 1H), 7.48 (d, J=8.0Hz, 2H), 7.39 (d, J=8.8Hz, 1H),7.38 (d, J=5.6Hz, 1H), 3.60 (s, 2H), 2.54 (br s, 8H), 2.40 (s, 3H), 2.32 (s, 3H).

Compound 5g:N-(4-methyl-3-((4-(pyridine-3-) pyrimidine-2-) amino) benzoyl) nicotinic acid hydrazide

Faint yellow solid, productivity 76%.

The MS:[M+H of this compound]⁺:426.01；¹HNMR(400MHz,DMSO-d₆)δppm 10.71(s, 1H), 10.53 (s, 1H), 9.27 (d, J=1.6Hz, 1H), 9.13 (s, 1H), 9.08 (d, J=1.6Hz,1H), 8.78 (dd, J=1.2,4.8Hz, 1H), 8.55 (d, J=5.2Hz, 1H), 8.47 8.43 (m, 1H),8.32 8.22 (m, 2H), 7.68 (dd, J=1.6,8Hz, 1H), 7.60-7.54 (m, 2H), 7.47 (d, J=5.2Hz, 1H), 7.40 (d, J=8Hz, 1H), 2.34 (s, 1H).

Compound 5h:N-acetyl group-4-methyl-3-((4-(pyridine-3-) pyrimidine-2-) amino) benzoyl hydrazine

Pale solid, productivity 87%.

The ESI-MS:[M+H of this compound]⁺:363.06；¹H NMR:10.21(s,1H),9.85(s,1H),9.26 (d, J=4Hz, 1H), 9.10 (s, 1H), 8.70 (dd, J=1.6,4.8Hz, 1H), 8.53 (d, J=5.2Hz, 1H), 8.43 (d, J=8Hz, 1H), 8.15 (s, 1H), 7.61 (dd, J=1.6,8Hz, 1H), 7.54 (dd,J=4.8,8Hz, 1H), 7.46 (d, J=5.2Hz, 1H), 7.36 (d, J=8Hz, 1H), 2.32 (s, 3H),1.92(s,3H)。

The synthesis of example 1-8 compound 6a-6h

Example 1 compound 6a:N-(2-methyl-5-{5-[3-(4-methyl-1 H-imidazole-1-group)-5-(trifluoromethyl) benzeneBase]-1,3,4-diazole-2-base } phenyl) synthesis of-4-(3-pyridine radicals)-pyrimidine-2-amine and Structural Identification

The triphen phosphine oxide adding 167mg in there-necked flask and the dichloromethane 2mL being dried, at 0 DEG CIt is slowly added to the trifluoromethanesulfanhydride anhydride of 50 μ L, at 0 DEG C, stirs 5min, and be slowly increased to room temperature, haveWhite solid generates, and adds compound 5a (114.5mg, 0.2mmol), and solution becomes glassy yellow, room10%NaHCO is used after temperature stirring reaction 3h₃Aqueous solution cancellation, water layer dichloromethane extracts, dichloroMethane layer anhydrous Na₂SO₄Being dried, filter, filtrate rotation, except solvent, residue column chromatography purification, is usedCH₂Cl₂: MeOH=50:1, CH₂Cl₂: MeOH=30:1 gradient elution, it is thus achieved that white solid 65mg,Productivity: 58.61%.

MS(ESI⁺): 554.98；HRMS-ESI (+): C₂₉H₂₁F₃N₈OH value of calculation 555.1869, realMeasured value 555.1854；

¹H NMR(400MHz,CDCl₃): δ ppm 9.38 (d, J=1.2Hz, 1H), 9.22 (d, J=1.2Hz, 1H), 8.65-8.66 (m, 1H), 8.59 (d, J=5.2Hz, 1H), 8.49-8.52 (m, 1H), 8.33-8.34(m, 2H), 7.93 (s, 1H), 7.85 (dd, J=1.6,7.6Hz, 1H), 7.78 (s, 1H), 7.41-7.44 (m,2H), 7.30 (d, J=5.2Hz, 1H), 7.19 (s, 1H), 7.14 (s, 1H), 2.50 (s, 3H), 2.35 (s, 3H)；

¹³C NMR(100MHz,CDCl₃)δppm165.8,162.6,162.4,160.3,159.3,151.7,148.5,140.7,138.7,138.4,134.5,134.4,133.8,133.5,132.3,132.1,131.4,126.9,124.2,123.8,121.7,121.7,121.7,120.0,119.3,114.2,108.8,18.4,13.6。

6b-6h is obtained according to above-mentioned route and method.

Example 2 compound 6b:N-(5-{5-[the fluoro-3-of 4-(trifluoromethyl)] phenyl }-1,3,4-diazole-2-methylbenzeneBase) Structural Identification of-4-(pyridin-3-yl) pyrimidine-2-amine

Yellowish white solid, productivity: 89.2%.

MS(ESI⁺): 493.33；HRMS-ESI (+): C₂₅H₁₆F₄N₆OH value of calculation 493.1400, actual measurementValue 493.1389；

¹H NMR(400MHz,CDCl₃) δ ppm9.37 (s, 1H), 9.20 (s, 1H), 8.73 (d, J=4Hz, 1H), 8.59 (d, J=5.2Hz, 1H), 8.52 (d, J=8.0Hz, 1H), 8.35-8.41 (m, 2H), 7.83 (d, J=7.6Hz, 1H), 7.37-7.47 (m, 3H), 7.17 (s, 1H), 2.49 (s, 3H)；

¹³C NMR(100MHz,CDCl₃)δppm 165.4,162.6,162.5,160.3,159.3,151.7,148.5,138.3,134.7,132.7,132.6,132.4,131.8,131.3,126.2,126.1,123.8,121.9,121.6,120.8,120.7,119.0,118.2,118.0,108.8,18.4。

Example 3 compound 6c:N-(5-{5-[3,5-bis-(trifluoromethyl) phenyl]-1,3,4-diazole-2-base }-2-methylPhenyl) Structural Identification of-4-(pyridin-3-yl) pyrimidine-2-amine

White solid, productivity: 64.9%.

MS(ESI⁺): 543.96；HRMS-ESI (+): C₂₆H₁₆F₆N₆OH value of calculation 543.1368, realMeasured value 543.1354；

¹H NMR(400MHz,CDCl₃)δppm9.35(s,1H),9.21(s,1H),8.71(s,1H),8.57 (s, 2H), 8.54 (s, 1H), 8.51 (d, J=7.6Hz, 1H), 8.05 (s, 1H), 7.81 (d, J=7.6Hz,1H),7.40-7.46(m,2H),7.29(m,1H),7.22(s,1H),2.48(s,3H)；

¹³C NMR(100MHz,CDCl₃)δppm 165.8,162.6,162.1,160.3,159.3,151.7,148.5,138.4,134.6,133.3,133.0,132.7,132.3,131.3,126.8,126.1,124.9,124.8,124.1,123.8,121.7,121.6,121.4,119.4,108.8,18.4。

Example 4 compound 6d:N-{2-methyl-5-[5-(4-methyl-3-trifluoromethyl)-1,3,4-diazole-2-base]Phenyl } Structural Identification of-4-(pyridin-3-yl) pyrimidine-2-amine

Obtain white solid, productivity=62%.

HRMS-ESI (+): C₂₆H₁₉F₃N₆OH value of calculation 489.1651, measured value 489.1644；

¹H NMR(400MHz,CDCl₃): δ ppm9.33 (s, 1H), 9.12 (s, 1H), 8.73 (d, J=0.4Hz, 1H), 8.58 (d, J=5.2Hz, 1H), 8.54 (d, J=8.0Hz, 1H), 8.35 (s, 1H),8.18 (d, J=7.6Hz, 1H), 7.82 (dd, J=1.6,8.0Hz, 1H), 7.39-7.46 (m, 3H), 7.18(s,1H),2.58(s,3H),2.47(s,3H)；

¹³C NMR(100MHz,CDCl₃): δ ppm 165.0,163.4,162.6,160.3,159.3,151.7,148.4,140.6,138.2,134.7,132.8,132.5,131.7,131.3,130.1,129.8,124.4,124.3,123.9,122.1,122.0,121.6,119.2,108.7,19.6,18.3。

Example 5 compound 6e:N-(2-methyl-5-{5-[3-(2-methyl-1 H-imidazole-1-group)-5-(trifluoromethyl) benzeneBase]-1,3,4-diazole-2-base } phenyl) Structural Identification of-4-(3-pyridine radicals)-pyrimidine-2-amine

Yellowish white solid, productivity: 60.6%.

MS(ESI⁺): 555.09；HRMS-ESI (+): C₂₉H₂₁F₃N₈OH value of calculation 555.1869, actual measurementValue 555.1857；

¹H NMR(400MHz,CDCl₃) δ ppm9.38 (d, J=1.6Hz, 1H), 9.21 (d, J=1.2Hz, 1H), 8.65-8.66 (m, 1H), 8.59 (d, J=5.2Hz, 1H), 8.47-8.50 (m, 1H), 8.46 (s,1H), 8.32 (s, 1H), 7.85 (dd, J=1.6,8.0Hz, 1H), 7.75 (s, 1H), 7.43-7.45 (m,2H), 7.30 (d, J=5.2Hz, 1H), 7.17 (s, 1), 2.50 (s, 3H), 2.47 (s, 3H)

¹³C NMR(100MHz,CDCl₃)δppm 165.8,162.6,162.3,160.3,159.3,151.7,148.5,144.6,139.5,138.4,134.5,133.6,133.3,132.2,131.4,128.5,126.7,126.6,124.8,124.7,124.1,123.8,123.2,121.7,121.7,121.4,120.4,119.3,108.8,18.4,13.9。

Example 6 compound 6f:N-(2-methyl-5-(5-(4-((4-methyl piperazine-1-) methyl) piperazine)-1,3,4-diazole-2-) phenyl) Structural Identification of-4-(pyridine-3-) pyrimidine-2-amine

White solid, productivity 63.3%.ESI-MS:[M+H]⁺:519.4；¹HNMR(400MHz,CDCl₃)δPpm 9.30 (d, 1H, J=1.2Hz), 9.13 (s, 1H), 8.70 (dd, 1H, J=1.2Hz, 4.8Hz),8.56 (d, 1H, J=5.2Hz) 8.53 (d, 1H, J=8Hz), 8.05 (d, 2H, J=8.4Hz), 7.80 (dd, 1H, J=1.6,8.0Hz), 7.46 (d, 2H, J=8.0Hz), 7.41 (dd, 1H, J=4.8,7.6Hz) 7.38 (d, 1H, J=8.4Hz),7.16(s,1H),3.58(s,2H),2.54(br,8H),2.45(s,3H),2.34(s,3H)。¹³C NMR(100MHz,CDCl₃):164.7,164.4,162.6,160.4,159.3,151.6,148.5,142.4,134.7,132.6,131.6,131.2,129.6,126.9,123.9,122.8,122.4,121.6,119.1,108.7,62.5,55.0,52.8,45.8,18.3。HR-MS(ESI)[M+H]⁺, value of calculation: 519.2615, measured value: 519.2560.

Example 7 compound 6g:N-(2-methyl-5-(5-(pyridine-3-)-1,3,4-diazole-2-) phenyl)-4-(pyridine-3-)The Structural Identification of pyrimidine-2-amine

Obtain white solid, productivity 64%.

The ESI-MS:[M+H of this compound]⁺:408.08；¹H NMR(400MHz,CDCl₃): δ ppm9.33 (d, J=6.8Hz, 2H), 9.16 (d, J=1.2Hz, 1H), 8.75 (dd, J=2.4,24.4Hz, 2H),8.56 (d, J=5.2Hz, 1H), 8.50 (d, J=8.4Hz, 1H), 8.41 (d, J=8Hz, 1H), 7.81 (dd,J=1.6,8Hz, 1H), 7.48 7.42 (m, 2H), 7.39 (d, J=8Hz, 1H), 7.18 (s, 1H), 2.46 (s,3H).¹³C NMR(100MHz,CDCl₃): 165.3,162.7,162.4,160.3,159.3,152.3,152.3,151.7,148.5,147.9,138.3,134.7,134.2,132.5,131.8,131.3,123.9,123.8,122.0,121.6,119.1,108.8,18.4。HR-MS(ESI)[M+H]⁺: value of calculation: 408.1573, measured value:408.1541。

Example 8 compound 6h:N-(2-methyl-5-(5-methyl isophthalic acid, 3,4-diazole-2-) phenyl)-4-(pyridine-3-) pyrimidine-2-The Structural Identification of amine

Faint yellow solid, productivity 65.3%.The ESI-MS:[M+H of this compound]⁺345.48；¹H NMR(400MHz,CDCl₃): δ ppm 9.33 (s, 1H), 9.11 (d, J=1.2Hz, 1H), 8.73 (d, J=3.6Hz,1H), 8.55 (d, J=5.2Hz, 1H), 8.48 (m, 1H), 7.70 (dd, J=1.6,8Hz, 1H) 7.46 (dd, J=4.4,7.6Hz, 1H), 7.35 (d, J=8Hz, 1H), 7.25 (d, J=5.2Hz, 1H), 7.13 (s, 1H),2.65(s,1H),2.44(s,1H)。¹³C NMR(100MHz,CDCl₃): δ ppm 165.1,163.5,162.6,160.3,159.2,151.6,148.6,138.1,134.7,132.5,131.2,131.0,123.7,122.5,121.3,118.8,108.7,18.2,11.2。HR-MS(ESI)[M+H]⁺, value of calculation: 345.1464, measured value:345.1446。

2.2.3.1 the synthesis of compound 8a-8d

Route and method are similar to the synthesis of compound 3.

2.2.3.1.1 the structure mirror of compound 8a:4-((4-methylpiperazine-1-yl) methyl)-3-(trifluoromethyl) benzoyl hydrazineFixed:

Faint yellow solid, productivity: 93.7%.

¹H NMR(400MHz,CDCl₃): δ ppm 8.04 (s, 1H), 7.95 (d, J=8.4Hz, 1H), 7.90(d, J=8.0Hz, 1H), 7.47 (s, 1H), 4.14 (s, 3H), 3.72 (s, 2H), 2.50-2.54 (m, 8H), 2.33(s,2H)。

2.2.3.1.2 compound 8b:4-{ [4-(2-tert-butyl diphenyl siloxy ethyl) piperazine-1-base] methyl }-3-(threeMethyl fluoride) Structural Identification of benzoyl hydrazine

Faint yellow solid, productivity: 98%.

¹H NMR(400MHz,CDCl₃): δ ppm 8.06 (s, 1H), 7.90-7.95 (m, 2H), 7.69-7.71(m, 4H), 7.38-7.48 (m, 6H), 4.16 (s, 2H), 3.84 (t, J=6.0Hz, 2H), 3.69 (s, 2H),2.51-2.65(m,10H),1.07(s,9H)。

2.2.3.1.3 compound 8c:4-[(N-t-butoxycarbonylaminoethyl)-4H-piperazine] methyl-3-(trifluoromethyl) benzene firstThe Structural Identification of hydrazides

Faint yellow solid, productivity: 79.3%.

¹H NMR(400MHz,CDCl₃):δppm 8.06(s,1H),7.90-7.95(m,2H),7.78(s,1H), 5.03 (s, 1H), 4.17 (s, 2H), 3.70 (s, 2H), 3.24 (d, J=4.8Hz, 2H), 2.48-2.51 (m,11H),1.47(s,9H)。

2.2.3.1.4 compound 8d:4-((4-(2-(tert-butyl diphenyl silica ethyl) piperazine-1-) methyl) benzoyl hydrazineStructural Identification

Pale yellow oily liquid body, productivity 92%.ESI-MS:[M+H]⁺:518.37.¹H NMR(400MHz,CDCl₃): δ ppm 7.70 7.65 (m, 6H), 7.43 7.33 (m, 9H), 4.09 (s, 2H), 3.80 (t, J=6Hz,2H),3.52(s,2H),2.59–2.45(m,10H),1.03(s,9H)。

2.2.3.1 the synthesis of compound 9a-9d

General routes outlined

2.2.3.2 the synthesis of compound 9a-9d

Route and method are similar to 5a-5h.

2.2.3.2.1 compound 9a:4-methyl-N'-{4-[(4-methylpiperazine-1-yl) methyl]-3-(trifluoromethyl) benzeneBase }-3-{ [4-(pyridine radicals-3-base) pyrimidine radicals-2-base] amino } Structural Identification of benzoyl hydrazine:

White solid, productivity: 78.3%.

¹H NMR(400MHz,CDCl₃): δ ppm 11.36 (s, 1H), 9.45 (d, J=1.6Hz, 1H),9.17 (s, 1H), 8.82 (dd, J=1.6,4.8Hz, 1H), 8.51 (d, J=5.2Hz, 1H), 8.29 (dd, J=1.6,6.4Hz, 1H), 8.23 (s, 1H), 8.19 (d, J=8.0Hz, 1H), 7.71 (d, J=8.0Hz, 1H),7.47 (dd, J=1.2,8.0Hz, 1H), 7.19-7.22 (m, 2H), 7.13 (s, 1H), 3.55 (s, 3H), 2.42(m,8H),2.38(s,3H),2.29(s,3H)。

2.2.3.2.2 compound 9b:4-{4-(2-tert-butyl diphenyl siloxy ethyl) piperazine-1-base] methyl }-N'-(4-Methyl-3-{ [4-(pyridine radicals-3-base) pyrimidine radicals-2-base] amino } phenyl) knot of-3-(trifluoromethyl) benzoyl hydrazineStructure is identified

White solid, productivity: 70.2%.

¹H NMR(400MHz,CDCl₃): δ ppm 11.55 (s, 1H), 11.34 (d, J=0.8Hz, 1H),9.42 (d, J=1.6Hz, 1H), 9.10 (s, 1H), 8.77 (dd, J=1.2,4.8Hz, 1H), 8.48 (d, J=5.2Hz, 1H), 8.23 (t, J=8.8Hz, 1H), 7.67-7.72 (m, 6H), 7.36-7.48 (m, 7H), 7.21 (s,1H),8.15-7.18(m,2H),3.80(s,2H),3.52(s,2H),2.34-2.57(m,13H),1.05(s,9H)。

2.2.3.2.3 compound 9c:4-[(N-t-butoxycarbonylaminoethyl)-4H-piperazine] methyl-N'-(4-methyl-3-{ [4-(pyridine radicals-3-base) pyrimidine radicals-2-base] amino } phenyl) Structural Identification of-3-(trifluoromethyl) phenylhydrazide

White solid, productivity: 96.1%.

¹H NMR(400MHz,CDCl₃):δppm 10.74(s,1H),10.53(s,1H),9.27(s,1H),9.17 (s, 1H), 8.69 (d, J=2.4Hz, 1H), 8.54 (d, J=4.8Hz, 1H), 8.44 (d, J=8.4Hz, 1H), 8.19-8.24 (m, 3H), 7.93 (d, J=7.6Hz, 1H), 7.67 (d, J=7.2Hz, 1H),7.55 (m, 1H), 7.46 (d, J=4.4Hz, 1H), 7.40 (d, J=8.0Hz, 1H), 3.54 (s, 2H), 3.20(d, J=1.2Hz, 2H), 2.33-2.42 (m, 13H), 1.46 (s, 9H).

2.2.3.2.4 compound 9d:N-(4-((4-(2-(tert-butyl diphenyl silica ethyl) piperazine-1-) methyl) benzoylBase) Structural Identification of-4-methyl-3-((4-(pyridine-3-) pyrimidine-2-) amino) benzoyl hydrazine

Pale solid, productivity 89%.ESI-MS:[M+H]⁺:805.29.¹HNMR(400MHz,DMSO-d₆): 10.43 (s, 3H), 9.27 (s, 1H), 9.12 (s, 1H), 8.70 (dd, J=1.2,4.4Hz, 1H),8.54 (d, J=5.2Hz, 1H), 8.46 (dd, J=1.6,8Hz, 1H), 8.22 (s, 1H), 7.89 (d, J=8Hz, 2H), 7.69 7.64 (m, 5H), 7.55 (dd, J=4.8,7.8Hz, 1H), 7.48 7.38 (m, 10H),3.73 (t, J=6Hz, 2H), 3.52 (s, 2H), 2.43 2.34 (m, 10H), 0.99 (s, 9H).

2.2.4 the synthesis of compound 10a-10i

Example 9 compound 10a:N-(2-methyl-5-{5-(4-[(4-methylpiperazine-1-yl) methyl]-3-(trifluoromethyl) benzeneBase }-1,3,4-diazole-2-base) synthesis of phenyl-4-(pyridin-3-yl) pyrimidine-2-amine and Structural Identification

Synthetic method is similar to 6a-6h with route.

Yellowish white solid, productivity: 85.3%.

MS(ESI⁺): 587.19；HRMS-ESI (+): C31H29F3N8OH value of calculation 587.2495,Measured value 587.2476；

¹H NMR(400MHz,CDCl₃) δ ppm 9.33 (s, 1H), 9.18 (d, J=1.2Hz, 1H),8.72 (d, J=3.6Hz, 1H), 8.59 (d, J=5.2Hz, 1H), 8.53 (d, J=4Hz, 1H), 8.37(s, 1H), 8.28 (d, J=8.4Hz, 1H), 8.01 (d, J=8.0Hz, 1H), 7.82-7.85 (m, 1H),7.41-7.46(m,2H),7.12(s,1H),3.76(s,2H),2.54-2.59(m,8H),2.49(s,3H),2.36(s,3H)；

¹³C NMR(100MHz,CDCl₃)δppm 165.1,163.4,162.7,160.4,159.3,151.7,148.5,141.8,138.3,134.6,132.4,131.8,131.3,131.2,129.9,129.7,124.4,124.3,123.8,122.8,122.1,121.6,119.2,108.8,58.0,55.2,53.1,46.0,18.4。

Example 10 compound 10b:2-(4-{4-[5-(4-methyl-3-{ [4-(pyridine radicals-3-base) pyrimidine radicals-2-base] amino }Phenyl]-1,3,4-diazole-2-base }-2-(trifluoromethyl) phenyl-peiperazinyl) synthesis of ethanol and Structural Identification

Compound 11 synthesizes: synthetic route is similar with 6a-6h with method.

Yellowish white solid 363mg, productivity: 70.2%

¹H NMR(400MHz,CDCl₃): δ ppm 9.34 (d, J=1.2Hz, 1H), 9.19 (d, J=1.2Hz, 1H), 8.72 (d, J=3.6Hz, 1H), 8.53-8.55 (m, 1H), 8.37 (s, 1H), 8.29 (d, J=8.0Hz, 1H), 8.00 (d, J=8.0Hz, 1H)), 7.84 (dd, J=1.6,7.6Hz, 1H), 7.69-7.71 (m,4H), 7.38-7.45 (m, 8H), 7.29 (d, J=5.2Hz, 1H), 7.17 (s, 2H), 3.87 (s, 2H), 3.75 (s,2H),2.41-2.66(m,13H),1.07(s,9H)。

Compound 10b synthesizes:

Compound 11 (363mg, 0.425mmol) and THF10mL room is added in 25mL there-necked flaskTemperature agitation and dropping enters TBAF (543.6mg, 2.123mmol), and room temperature reaction, after 3 hours, adds saturatedSodium bicarbonate aqueous solution, extracts with dichloromethane, and dichloromethane layer anhydrous sodium sulfate is dried, and filters, filterLiquid revolves with Rotary Evaporators and removes solvent, residue column chromatography purification, it is thus achieved that white solid 34b, 240mg,Productivity: 91.7%.

MS(ESI⁺): 617.12；HRMS-ESI (+): C₃₂H₃₁F₃N₈O₂H value of calculation 617.2600, realMeasured value 617.2548；

¹H NMR(400MHz,CDCl₃) δ ppm9.32 (s, 1H), 9.16 (s, 1H), 8.71 (d, J=4Hz, 1H), 8.57 (d, J=5.2Hz, 1H), 8.52 (d, J=8.0Hz, 1H), 8.36 (s, 1H), 8.28 (s,1H), 8.27 (d, J=4.4Hz, 1H), 7.98 (d, J=8.4Hz, 3H), 7.82 (d, J=8.0Hz, 1H),7.39-7.44 (m, 2H), 7.28 (s, 1H), 7.22 (s, 1H), 3.75 (s, 3H), 3.69 (t, J=5.2Hz,3H),2.61-2.66(m,10H),2.47(s,3H)；

¹³C NMR(100MHz,CDCl₃)δppm 165.1,163.3,162.6,160.3,159.3,151.7,148.5,141.6,138.3,134.6,132.4,131.9,131.3,131.2,129.9,129.8,129.4,125.1,124.4,124.3,123.8,122.9,122.4,122.0,121.609,119.226,108.7,59.4,57.6,53.4,53.0,18.4。

Example 11 compound 10c:N-(2-methyl-5-{5-(4-[(4-aminoethyl piperazine-1-base) methyl]-3-(trifluoromethyl)Phenyl }-1,3,4-diazole-2-base) synthesis of phenyl-4-(pyridin-3-yl) pyrimidine-2-amine and Structural Identification

Compound 12:N-(2-methyl-5-{5-(4-[(N-t-butoxycarbonylaminoethyl)-4H-piperazine methyl]-3-(fluoroformBase) phenyl }-1,3,4-diazole-2-base) phenyl-4-(pyridin-3-yl) pyrimidine-2-amine

Synthetic route is similar with 6a-6h with method.

Yellowish white, productivity: 74.9%.

¹H NMR(400MHz,CDCl₃): δ ppm 9.31 (s, 1H), 9.13 (s, 1H), 8.69 (d, J=2.8Hz, 1H), 8.55 (d, J=4.8Hz, 1H), 8.50 (d, J=8.0Hz, 1H), 8.34 (s, 1H), 8.24 (d,J=8.0Hz, 3H), 7.96 (d, J=8.0Hz, 1H), 7.79 (d, J=7.6Hz, 1H), 7.36-7.42 (m,2H),7.24-7.28(m,2H),5.18(s,1H),3.73(s,2H),3.29(s,2H),2.59(m,10H),2.44(s,3H),1.46(s,9H)。

Compound 10c:N-(2-methyl-5-{5-(4-[(4-aminoethyl piperazine-1-base) methyl]-3-(trifluoromethyl) benzeneBase }-1,3,4-diazole-2-base) phenyl-4-(pyridin-3-yl) pyrimidine-2-amine

Compound 12 (738.9mg, 1.03mmol), CHCl is added in 25mL there-necked flask₃(3mL)And CH₃OH (3mL), is stirred at room temperature, and drips concentrated hydrochloric acid 4mL, continues stirring 30 minutes, usesSaturated sodium carbonate regulation ph to 11, extracts with dichloromethane and methanol (10:1) mixed liquor, and organic layer is usedAnhydrous sodium sulfate is dried, and filters, and filtrate rotation, except solvent, residue column chromatography purification, obtains white-yellowish solid482.1mg, productivity: 75.86%.

MS(ESI⁺): 616.71；HRMS-ESI (+): C₃₂H₃₂F₃N₉OH value of calculation 616.2760, actual measurementValue 616.2746；

¹H NMR(400MHz,CDCl₃): δ ppm 9.33 (d, J=1.6Hz, 1H), 9.18 (s, 1H),8.72-8.73 (m, 1H), 8.59 (d, J=5.2Hz, 1H), 8.54 (d, J=8.0Hz, 1H), 8.37 (s,1H), 8.28 (d, J=8.0Hz, 1H), 8.02 (d, J=8.0Hz, 1H), 7.83 (d, J=8.0Hz, 1H),7.41-7.45 (m, 2H), 7.28 (s, 1H), 7.17 (s, 1H), 3.75 (s, 2H), 2.83 (t, J=6.0Hz, 2H),2.56(br,8H),2.47-2.50(m,5H)；

¹³C NMR(100MHz,CD₃OD)δppm 165.10,163.19,161.87,160.51,159.14,150.51,147.53,141.72,138.54,135.19,134.03,132.62,131.38,131.06,129.76,(128.99,129.76,129.30 q, J=30.9Hz), 124.04,108.02,59.69,57.67,53.01,52.69,37.54,17.01。

Example 12 compound 10d:N-(5-{5-(4-[(4-(2-isothiocyanic acid aliphatic radical) ethyl piperazidine-1-base) methyl]-3-(threeMethyl fluoride) phenyl } synthesis of-1,3,4-diazole-2-aminomethyl phenyl-4-(pyridin-3-yl) pyrimidine-2-amine and structureIdentify

Under nitrogen protection, 25mL there-necked flask puts into addition dichloromethane 10mL at 0 DEG C, stirs 5 minutes,Be subsequently adding compound 10c (200mg, 0.325mml), DCC (73.7mg, 0.357mmol) andCS₂(196.3uL, 3.25mmol), moves to room temperature from 0 DEG C after feeding intake, after reaction 3h, with rotationTurn evaporimeter rotation except reactant liquor solvent, residue column chromatography purification (CH₂Cl₂: MeOH=30:1) must be newbornWhite solid 192.6mg, productivity: 90.1%.

MS(ESI⁺)658.41；HRMS-ESI (+): C₃₃H₃₀F₃N₉OSH value of calculation 658.2324, actual measurementValue 658.2317；

¹H NMR(400MHz,CDCl₃): δ ppm 9.33 (d, J=1.2Hz, 1H), 9.18 (d, J=1.2Hz, 1H), 8.72 (d, J=3.6Hz, 1H), 8.59 (d, J=5.2Hz, 1H), 8.53-8.55 (m, 1H), 8.37(s, 1H), 8.28 (d, J=8.0Hz, 1H), 8.00 (d, J=8.0Hz, 1H), 7.83 (dd, J=2.4,8.0Hz,1H), 7.40-7.45 (m, 2H), 7.30 (s, 1H), 7.17 (s, 1H), 3.76 (s, 2H), 3.63 (t, J=6.0Hz, 2H), 2.73 (t, J=6.0Hz, 2H), 2.60 (br, 8H), 2.8 (s, 3H)；

¹³C NMR(100MHz,CDCl₃)δppm 165.14,163.34,162.65,160.35,159.30151.72,148.52,141.73,138.28,134.63,132.43,131.82,131.30,131.17,129.94,129.61 (q, J=30.5Hz), 124.38,124.32,123.83,123.78 (q, J=272.4Hz),122.87,122.10,121.62,119.21,108.78,57.93,57.21,53.12,52.99,43.07,18.36。

Example 13 compound 10e:N-(5-{5-(4-[(4-(2-acrylamide ethyl piperazidine-1-base) methyl]-3-(fluoroformBase) phenyl } synthesis of-1,3,4-diazole-2-aminomethyl phenyl-4-(pyridin-3-yl) pyrimidine-2-amine and Structural Identification

At room temperature, 10mL there-necked flask adds compound 10c (50mg, 0.081mmol), EDCI(31.3mg, 0.163mmol), HOBT H₂O (24.9mg, 0.163mmol), NMM (27.5uL,After 0.244mmol) stirring 20 minutes with dichloromethane 5mL, add acrylic acid (8.5ul, 0.122mmol),Reaction 24 was as a child adjusted to alkalescence with saturated sodium bicarbonate, extracted with dichloromethane, dichloromethane layer nothingAqueous sodium persulfate is dried, and filters, with Rotary Evaporators rotation except filtrate solvent, residue column chromatography purification, it is thus achieved thatWhite solid 22.5mg, productivity: 41%.

HRMS-ESI (+): C₃₅H₃₄F₃N₉O₂H value of calculation 670.2866, measured value 670.2869；

¹H NMR(400MHz,CDCl₃): δ ppm 9.29 (s, 1H), 9.12 (s, 1H), 8.67 (d, J=4.0Hz, 1H), 8.54 (d, J=5.2Hz, 1H), 8.49 (d, J=8.0Hz, 1H), 8.33 (s, 1H), 8.23 (d, J=8.0Hz, 1H), 7.96 (d, J=8.4Hz, 1H), 7.78 (d, J=7.6Hz, 1H), 7.35-7.41 (m, 2H),7.25 (d, J=5.2Hz, 1H), 7.21 (s, 1H), 6.38 (s, 1H), 6.24 (d, J=16.8Hz, 1H), 6.13(dd, J=10.0,16.8Hz, 1H), 5.62 (d, J=10.0Hz, 1H), 3.71 (s, 2H), 3.45 (dd, J=5.2,10.8Hz,2H),2.55-2.58(br,10H),2.43(s,3H)；

¹³C NMR(100MHz,CDCl₃)δ165.54,165.13,163.30,162.61,160.37,159.25,151.64,148.49,141.62,138.30,134.62,132.44,131.94,131.26,131.18,130.98,129.93,129.71,129.40,126.12,125.15,124.35,124.29,123.80,122.90,122.43,122.05,121.62,119.29,108.71,57.92,56.42,53.04,52.90,35.89,18.31。

Example 14 compound 10f:2-(4-(4-(5-(4-methyl-3-((4-(pyridine-3-) pyrimidine-2-) amino) phenyl)-1,3,4-Diazole-2-) benzyl) piperazine-1-) synthesis of ethanol (A15) and Structural Identification

Compound 13 synthesizes: synthetic route is similar with 6a-6h with method.

Faint yellow solid, productivity 67%.ESI-MS:[M+H]⁺:786.91.¹H NMR(400MHz,CDCl₃): 9.32 (s, 1H), 9.12 (s, 1H), 8.72 (s, 1H), 8.56 (d, J=4.8Hz, 1H), 8.51 (d, J=8Hz, 1H), 8.08 (d, J=8Hz, 2H), 7.80 (dd, J=1.2,7.6Hz, 1H), 7.64 7.62 (m,5H),7.48–7.38(m,11H),7.18(s,1H),4.04(s,2H),3.68(s,2H),3.12–2.82(m,11H),2.46(s,3H),1.04(s,9H)。

Compound 10f synthesizes: synthetic route is similar with 10b with method.

Faint yellow solid, productivity 94%.ESI-MS:[M+H]⁺:549.08；¹H NMR(400MHz,CDCl₃): δ ppm 9.28 (s, 1H), 9.11 (s, 1H), 8.70 (d, J=3.6Hz, 1H), 8.55 (d, J=5.2Hz,1H), 8.52 (d, J=8Hz, 1H), 8.05 (d, J=8Hz, 2H), 7.80 (d, J=8Hz, 1H), 7.45 (d,J=8Hz, 2H), 7.39 (m, 2H), 7.17 (s, 1H), 3.66 (t, J=5.2Hz, 2H), 3.58 (s, 2H),2.63-2.55(m,10H),2.44(s,3H)。¹³C NMR(100MHz,CDCl₃):δppm 164.7,164.4,162.6,160.3,159.3,151.6,148.5,142.2,138.2,134.7,132.6,131.6,131.2,129.6,127.0,123.9,122.8,122.4,121.6,119.2,108.7,62.5,59.4,57.6,52.9,18.3。HR-MS(ESI)[M+H]⁺, value of calculation: 549.2726, measured value: 549.2715.

Example 15 compound 10g:N-(5-(5-(4-((4-(2-aminoethyl) piperazine-1-) methyl) phenyl)-1,3,4-diazole-2-)-2 aminomethyl phenyls) synthesis of-4-(pyridine-3-) pyrimidine-2-amine and Structural Identification

The synthesis of compound 14

In two mouthfuls of flasks of 25mL, adding compound 10f (50mg, 0.09mmol), phthalyl is sub-Amine (17.4mg, 0.11mmol), triphenylphosphine (31mg, 0.11mmol), the oxolane that 5mL is dried.Said mixture is placed in ice bath and is cooled to-5 DEG C, be slowly added dropwise in above-mentioned solution DEAD (19 μ L,0.11mmol).After dropping, reactant liquor is moved to room temperature, be stirred overnight, and by TLC to reactionIt is monitored.After reaction completely, concentrating under reduced pressure, add 20mL dichloromethane, organic facies sodium carbonateSolution and saturated aqueous common salt washing, and be dried with anhydrous sodium sulfate, concentrating under reduced pressure, and pass through silica gel column chromatography(dichloromethane: methanol=40:1) carries out isolated and purified, it is thus achieved that white solid, productivity 74%.

The MS:[M+H of this compound]⁺:678.21.¹HNMR:(400MHz,CDCl₃)δppm 9.30(s,1H), 9.14 (s, 1H), 8.72 (s, 1H), 8.58 (d, J=5.2Hz, 1H), 8.55 (d, J=8Hz, 1H),8.05 (d, J=6.8Hz, 2H), 7.86 7.81 (m, 3H), 7.73 7.71 (m, 2H), 7.44 7.39 (m,4H), 7.10 (s, 1H), 3.82 (t, 2H, J=6.4Hz), 3.55 (s, 2H), 2.66 2.47 (m, 13H).

The synthesis of compound 10g

In two mouthfuls of flasks of 25mL, add compound 14 (80mg, 0.12mmol), 1mL hydrazine hydrate and5mL ethanol, reactant liquor is stirred at room temperature overnight, and is monitored by TLC.Subtract after having reactedPressure concentrates, and adds 20mL 5% sodium carbonate liquor, and aqueous phase dichloromethane extracts, and merges organic facies,And be dried with anhydrous sodium sulfate, concentrating under reduced pressure, and carry out isolated and purified (dichloro by silica gel column chromatographyMethane: methanol=10:1 to 5:1+5 ‰ ammonia), it is thus achieved that faint yellow solid, productivity 35%.ESI-MS:[M+H]⁺:547.77.¹HNMR:(400MHz,DMSO-d₆) δ ppm 9.33 (d, 1H, J=2Hz), 8.68(dd, 1H, J=1.6,4.8Hz), 8.60 (d, 1H, J=5.2Hz), 8.55 (s, 1H), 8.50 (d, 1H, J=8Hz), 8.03 (d, 2H, J=8Hz), 7.82 (d, 1H, J=8Hz), 7.55 7.50 (m, 5H), 3.55 (s,2H), 2.64 (t, 2H, J=6.8Hz), 2.40 (m, 8H), 2.32 (t, 2H, J=7.2Hz).¹³C NMR(100MHz,DMSO-d₆):δppm 164.5,164.3,162.0,161.2,160.2,151.9,148.6,143.2,139.3,136.2,134.8,132.6,132.5,131.9,130.0,128.4,127.0,125.7,124.3,122.4,122.3,121.6,108.8,62.04,59.50,53.27,53.10,38.20,18.79。HR-MS(ESI)[M+H]⁺, value of calculation: 548.2886, measured value: 548.2886.

Example 16 compound 10h:(1E, 6E)-1-(hydroxyl-3-anisyl)-7-(3-methoxyl group-4-(2-(4-4-(5-4-Methyl-3-((4-pyridine-3-) pyrimidine-2-) amino) phenyl)-1,3,4-diazole-2-) benzyl) piperazine-1-) ethyoxyl)The synthesis of phenyl 1,6-heptadiene-3,5-diketone and Structural Identification

In the there-necked flask that 25mL is dried, add 50mg compound 10f (50mg, 0.1mmol), changeCompound 39 (43.7mg, 0.12mmol), triphenylphosphine (31.1mg, 0.14mmol) and oxolane (5ML), above-mentioned solution ice bath is cooled to-5 DEG C, is under agitation added drop-wise to by DEAD (19 μ L, 0.14mmol)In above-mentioned solution, after dropping, reactant liquor is warming up to room temperature, be stirred overnight and pass through TLC to react intoRow monitoring, reaction terminates rear concentrating under reduced pressure and carries out isolated and purified (dichloromethane: first by silica gel column chromatographyAlcohol=40:1 to 15:1), it is thus achieved that orange solids, productivity 24%.ESI-MS:[M+H]⁺:899.38.¹H NMR(400MHz,DMSO-d₆): δ ppm 9.66 (s, 1H), 9.32 (s, 1H), 9.15 (s, 1H), 8.68 (s, 1H),8.59 (d, J=2.8Hz, 1H), 8.55 (s, 1H), 9.45 (d, J=6.4Hz, 1H), 8.03 (d, J=6.8Hz,2H), 7.81 (d, J=6.4Hz, 1H), 7.58 7.49 (m, 7H), 7.33 (d, J=2Hz, 2H), 7.23 (d,J=6.4Hz, 1H), 7.15 (d, J=6.8Hz, 1H), 7.02 (dd, J=0.4,6.4Hz, 1H), 6.836.74(m,3H),6.07(s,1H),4.11(s,2H),3.84(s,3H),3.82(s,3H),3.55(s,2H),2.72(s,2H),2.39(s,7H)。¹³C-NMR(100MHz,DMSO-d₆): δ ppm 184.2,183.3,183.2,164.5,164.3,162.1,161.2,160.2,152.0,150.6,149.9,149.7148.6,148.5,143.3,141.4,140.6,139.3,136.2,134.8,132.6,132.0,130.0,128.2,127.1,126.8,124.3,123.6,123.3,122.6,122.5,122.4,121.6,121.6,116.2,113.4,111.9,111.3,108.8,101.4,66.8,62.0,57.0,56.2,53.6,53.1,18.8。HR-MS(ESI)[M+H]⁺, value of calculation:899.3881, measured value: 899.3793.

Example 17 compound 10i:N-(5-(5-(4-((4-(2-isothiocyanic acid ethyl) piperazine-1-) methyl) phenyl)-1,3,4-Diazole-2-)-2-aminomethyl phenyl) synthesis of-4-(pyridine-3-) pyrimidine-2-amine and Structural Identification

Synthetic route is similar with 10d with method.

Faint yellow solid, productivity 20%.The ESI-MS:[M+H of this compound]⁺: 590.18,¹HNMR:(400MHz,CDCl₃): δ ppm 9.28 (d, J=2Hz, 1H), 9.11 (d, J=0.8Hz, 1H), 8.70 (d, J=1.6,4.8Hz, 1H), 8.56 (d, J=5.2Hz, 1H), 8.53 (d, J=8Hz, 1H), 8.15 (d, J=8Hz,2H), 7.81 (dd, J=1.2,7.6Hz, 1H), 7.46 (d, J=8Hz, 2H), 7.42 7.38 (m, 2H),7.25 (s, 1H), 7.14 (s, 1H), 3.59 (t, J=6.4Hz, 4H), 2.69 (t, J=6.4Hz, 2H), 2.572.53(m,8H),2.44(s,3H).¹³C NMR(100MHz,CDCl₃): δ ppm 164.7,164.4,162.6,160.4,159.3,151.6,148.5,138.2,134.7,132.6,131.6,131.2,129.6,126.9,123.9,122.9,122.4,121.6,119.2,108.7,62.5,57.2,53.1,52.8,43.1,29.7,18.3.HR-MS(ESI)[M+H]⁺, value of calculation: 590.2451, measured value: 590.2438.

The active testing of example 1 to example 17 compound that embodiment 2 prepares in embodiment 1

MTT powder is bought in Sigma company, and being configured to concentration with phosphate buffer (PBS) is 5 millisThe solution of grams per milliliter, chooses 0.22 μm filter membrane and filters, then keep in Dark Place at 4 DEG C.K562Cell (chronic myeloid leukemia cell line) is purchased from from gold amethyst bio tech ltd, Beijing.ExperimentUsed in cell culture medium be modified form RPMI-1640 (Hyclone) basal medium, add 10%Hyclone (Hyclone).

MTT colorimetric method for determining cytoactive includes that following several step (with the method for testing of K562 cell isExample, the method for testing of KG1a HL60 cell is consistent with the method for K562 cell):

(1) choose and be in the K562 cell of exponential phase, by every hole 3 × 10³It is inoculated in 96 orifice plates,5%CO₂, hatch overnight incubation for 37 DEG C.

(2) being provided with 9 Concentraton gradient in dosing, this experiment, system Chinese medicine final concentration gradient is:30 μMs, 15 μMs, 7.5 μMs, 3.7 μMs, 1.8 μMs, 0.9 μM, 0.5 μM, 0.2 μM, 0.1 μM.The multiple hole of each concentration 5, arranges matched group (not dosing only inoculating cell) simultaneously and blank well (is not inoculatedCell only adds culture medium), 5%CO₂, 37 DEG C of incubators hatch 72 hours.

(3) every hole adds 20 μ L MTT solution (5mg/ml, i.e. 0.5%MTT), continues to cultivate 4 littleTime.If medicine can react with MTT, can first be centrifuged and discard culture fluid afterwards, carefully rinse 2-3 with PBSAfter Bian, add the culture fluid containing MTT.

Terminate after (4) 4 hours cultivating, suck the liquid in hole carefully.And 150 μ L are added to every holeDimethyl sulfoxide.It is subsequently placed in low-speed oscillation about 15min on shaking table, makes crystal fully dissolve.AdoptMeasure at 490nm with enzyme-linked immunosorbent assay instrument MULTISKAN FC (Thermo scientific) andThe absorbance in each hole of 570nm, using blank well as zeroing hole during measurement.

(5) data are processed.With drug level as abscissa, cell number is vertical coordinate, processes soft by dataPart SPSS software (IBM Corporation) carries out probit weighted regression method (Bliss method) and carries out data process,Mapping, obtains IC₅₀And IC₉₀Value, IC₅₀And IC₉₀It is shown in Table 1.

The table 1 example 1-17 compound IC to K562-3d, HL60, KG1a cell₅₀And IC₉₀Value

By table 1, it can be seen that example 1 to example 17 compound prepared in an embodiment is (the most rightShould be in compound 6a-6h and 10a-10i) all can effectively suppress K562-3d, HL60, KG1a cellPropagation, particularly example 11 compound (compound 10c) at drug level less than positive control medicineIn the case of imatinib, still can realize equally effectively suppressing K562-3d, HL60, KG1a cell to increaseThe effect grown, therefore, example 1 to example 17 compound that the present invention provides may be used for treating leukemia.

The foregoing is only presently preferred embodiments of the present invention, not in order to limit the present invention, allWithin the spirit and principles in the present invention, any modification, equivalent substitution and improvement etc. made, all should wrapWithin being contained in protection scope of the present invention.

Claims (10)

1. a heterocycle compound Han oxadiazoles, it is characterised in that there is following general structure:

The most according to claim 1 containing oxadiazoles heterocycle compound, it is characterised in that describedIn formula, R includes following group:

3. the preparation method containing oxadiazoles heterocycle compound, it is characterised in that include following conjunctionOne-tenth route:

Make N-acyl group-4-methyl-3-((4-(pyridine-3-) pyrimidine-2-) amino) benzoyl hydrazine compound that weight to occurRow's reaction, is converted into the described heterocycle compound Han oxadiazoles；

Wherein, described N-acyl group-4-methyl-3-((4-(pyridine-3-) pyrimidine-2-) amino) benzoyl hydrazine chemical combinationThing has a following general structure:

Further,

The described heterocycle compound containing oxadiazoles has a following general structure:

Preparation method the most according to claim 3, it is characterised in that described N-acyl group-4-firstThe general structure of base-3-((4-(pyridine-3-) pyrimidine-2-) amino) benzoyl hydrazine compound and described containing evil twoIn the general structure of azoles heterocycle compound, R includes following group:

Preparation method the most according to claim 3, it is characterised in that at Trifluoromethanesulfonic anhydrideUnder conditions of existing with triphenylphosphine oxide or under conditions of potassium carbonate and paratoluensulfonyl chloride exist, send outRaw described rearrangement reaction.

Preparation method the most according to claim 4, it is characterised in that described N-acyl group-4-firstThe preparation method of base-3-((4-(pyridine-3-) pyrimidine-2-) amino) benzoyl hydrazine compound farther includes:1-(3-dimethylamino-propyl)-3-ethyl-carbodiimide hydrochloride (EDCl), I-hydroxybenzotriazole(HOBT) and in the presence of N-methylmorpholine (NMM), 4-methyl-3-{ [4-(pyridine radicals-3-base) is madePyrimidine radicals-2-base] amino } benzoyl hydrazine reacts with carboxylic acid compound (R-CH3COOH), generates describedN-acyl group-4-methyl-3-((4-(pyridine-3-) pyrimidine-2-) amino) benzoyl hydrazine compound；

Wherein, R includes following group:

7. according to controlling being used for containing oxadiazoles heterocycle compound according to any one of claim 1-6Treat the application in leukemic medicine.

Application the most according to claim 7, it is characterised in that described leukemic for treatingPharmaceutical pack containing described containing oxadiazoles heterocycle compound, the described heterocycle compound containing oxadiazoles in pharmacyUpper acceptable salt, ester, hydrate or combinations thereof and adjuvant.

Application the most according to claim 7, it is characterised in that the leukemic medicine of described treatmentDosage form selected from tablet, capsule, pill, suppository, aerosol, oral liquid, granule,Powder, injection, syrup, medicated wine, tincture, distillate medicinal water, membrane or combinations thereof.

Application the most according to claim 7, it is characterised in that the leukemic medicine of described treatmentThe administering mode of thing includes being administered orally, injects, implants, external, spray, suck or combinations thereof.