CN107596394A - Using full-automatic, one-step synthesis method18The method of the pyrimidine acrylic amide EGFR positive electron tracers of F marks - Google Patents

Abstract

The invention discloses using full-automatic, one-step synthesis method¹⁸The method of the pyrimidine acrylic amide EGFR positive electron tracers of F marks, belongs to the field of chemical synthesis.Methods described can be carried out successively as follows：1) accelerator will be come from¹⁸F ion and water pass to receiving bottle and imported into anion-exchange column；2) using the mixed solution of strong base-weak acid salt, acetonitrile and ionic liquid by the anion-exchange column¹⁸F ion is eluted and is sent to reaction bulb；3) precursor is dissolved in DMSO, acetonitrile or DMF solution, adds cushioning liquid to make pH value of solution=2, solution is added in the reaction bulb and heated under the protection of inert gas：4) temperature is reduced, the crude product in the reaction bulb is purified by SPE C18 pillars；5) target product to step 4) resulting solution is separated, collected, and is produced.The present invention possesses the features such as big multipurpose, yield, efficiency high, short low with cost generated time, disclosure satisfy that the needs of large-scale production.

Description

Using full-automatic, one-step synthesis method18The pyrimidine acrylic amide EGFR of F marks is just The method of electronics tracer

Technical field

The present invention relates to a kind of by being fluorinated substitution reaction synthesis¹⁸The pyrimidine acrylic amide EGFR positive electrons of F marks Tracer and preparation method thereof, the invention belongs to the field of chemical synthesis,

Background technology

EGF-R ELISA (EGFR, ErbB) is the thin of the extracellular protein part of I skins growth factor family Cellular surface acceptor, there is tyrosine kinase activity, be a kind of important transmembrane receptor.EGFR high expression can promote tumour thin The propagation of born of the same parents, angiogenesis, stick, attack and shift, therefore EGFR tyrosine kinase inhibitors become antineoplastic and ground The focus studied carefully.Research thinks that the mutation in EGFR gene T790M sites is the main inducing of such Drug-resistant, existing in order to overcome The resistance reason of medicine, herein using EGFR inhibitor AZD-9291 as lead compound, carry out structural modification, it is intended to find activity The good, EGFR inhibitor selectively high to drug-resistant tumor.Between the decade of past two, with a large amount of clinical trials and research, with The micromolecular inhibitor of EGFR receptor site target spots just with the characteristics of its high selectivity, hypotoxicity and high therapeutic index by medical work The attention of author.EGFR targeted therapies are compared with classic chemotherapy, because having more preferable security and tolerance, in non-small cell lung Occupy increasingly consequence in the treatments of malignant tumour such as cancer (NSCLC), but medical expense is quite high, therefore treat Preceding clear and definite EGFR expression, treatment and Index for diagnosis and medicine health economics to tumour have certain value.With The extensive clinical practice of EGFR molecular targeted therapies, EGFR molecular imagings, which also gradually cause, pays close attention to and obtained preliminary grind Study carefully.

It is the pith of tumor cells targeted therapy using EGFR as therapy target, mainly there is monoclonal antibody and small molecule Tyrosine kinase antagonist two ways, such as Cetuximab (Cetuximab, Erbitux, Erbitux), Buddhist nun's trastuzumab (Nimotuzumab, nimotuzomab) and Gefitinib (Gefitinib, Iressa, Iressa), Tarceva (Erlotinib, Tarceva, Erlotinib) etc. preferable therapeutic effect is shown in clinical trial and clinical practice.But nearly all patient exists Inevitably there is resistance in 1 year or so in medication.Therefore, the molecular mechanism and its regulation measure of resistance are furtherd investigate, will be favourable In the clinical therapeutic strategy found for different resistance mechanisms, and then improve the life cycle of patient.In November, 2015, U.S. FDA was criticized Accurate IIIth generation EGFR-TKI osimertinib (AZD9291) treatment EGFR T790M are mutated or to other EGFR inhibitor resistances Advanced NSCLC patients, enable about 50% EGFR-TKI resistance patients to continue to benefit from targeted therapy, turn into EGFR-TKI The model that therapeutic scheme is explored after resistance.

(1) third generation small molecule EGFR acceptor inhibitors and its application

1st, third generation small molecule EGFR acceptors and its classification

Epidermal growth factor recipient tyrosine kinase family is also referred to as erbB receptor families, including erbB-1 (EGFR/HER1), Tetra- members of erbB-2, erbB-3, erbB-4.EGF-R ELISA (epidermal growth factor Receptor, EGFR) it is the common cancer driving genes of NSCLC.Epidermal growth factor recipient tyrosine kinase inhibitor (epidermal growth factor receptor tyrosine kinase inhibitor, EGFR-TKI) can be with The ATP site competitions of EGFR tyrosine kinase domains combine, and suppress its activation and phosphorylation, are passed under disabling signal, so as to press down The propagation of tumour cell processed, promote apoptosis of tumor cells, finally suppress tumour growth.According to molecular structure and mechanism of action not Together, EGFR-TKIs can be divided into a generation, two generations and three generations TKIs again, although the advanced NSCLC patients of EGFR mutation receive EGFR- The effective percentage of TKIs treatments can reach 60%~80%, but Most patients finally all can inevitably occur to EGFR-TKI Resistance.But for the resistance mechanism of EGFR medicament-resistant mutations, still there are many uncertain parts, so understanding EGFR-TKIs in depth Resistance mechanism, to TKIs individualized treatment, Index for diagnosis, early diagnosis resistance patient, delay or reversing drug resistance has emphatically The meaning wanted.

2nd, third generation small molecule EGFR inhibitor is in clinical practice

Third generation EGFR micromolecular inhibitors, which represent medicine, mainly includes HM61713, CO1686 and AZD9291.

HM61713 is that a kind of new third generation has Orally active, irreversible, selective is mutated for EGFR Tyrosine kinase inhibitor.In American Society of Clinical Oncology (ASCO) annual meeting held in 2014, I/II phases clinic is described The term results of experiment, and show the effect of strong signal and favourable security features.The medicine is currently in the II phases and faced The bed development phase.Plan also to have begun in the preparation of the III phase pilot projects started in 2016.

CO1686 is a kind of new, oral, irreversible covalent EGFR inhibition from mutation agent of targeting, can suppress crucial activation Mutation and T790 medicament-resistant mutations, make Wild type EGFR signal leave unused.On May 20th, 2014, Clovis tumour company announce, beautiful State FDA authorizes the breakthrough medicine qualifications of its trial drug CO1686, and it dashes forward as single medicine two wires for treating T790M Become patient EGFR mutation non-small cell lung cancers.The breakthrough medicine qualification is authorized based on CO-1686 ongoing one The effectiveness and reliability result of 1/2 phase of item research mid-term.

AZD-9291 is that the third generation is oral, irreversible selective EGFR inhibition from mutation agent, available for activating resistant mutation EGFR.That is, for patients with advanced NSCLC, 50% anti-EGFR treatment acquired resistances are by T790M Caused by mutation.AZD9291 for Wild type EGFR effect very little, it is therefore smaller than existing EGFR inhibitor side effect. AZD9291 is using oral administration, 20-240mg/ days.Brain metastes (being in stable state) patient is also included into the research.All doses Horizontal pharmacokinetics, therapeutic response and the toxic reaction that medicine will be observed with Asia group crowd of amount.199 EGFR included It is mutated in advanced NSCLC patients, 51% tumor regression.In 89 T790M mutation patients, 64% patient has instead to AZD9291 Should, do not find have with dose-dependent toxicity, most common adverse reaction：Suffer from diarrhoea (30%), erythema (24%), nausea (17%).Present EGFR TKI medicines can block the normal EGFR in the EGFR and skin of Tumor mutations simultaneously, typically result in Serious fash or acne, and AZD9291 mainly acts on the mutation EGFR of tumour, is caused less than other EGFR TKI medicines Dermal toxicity.Statistics by when, nearly all patient for treatment still has reaction, most long to continue 8 wheat harvesting periods.

(2) whether detection EGFR is mutated, screens small molecule EGFR inhibitor beneficiaries and monitoring treatment curative effect at present Method and its limitation

At present, may predict EGFR targeted therapy curative effects biological markers include the high expression of EGFR albumen or mRNA, EGFR gene copy number, EGFR genetic mutation, K-RAS gene mutations, the Research of predicting markers etc. of EGFR downstream signal systems.But still Recommend applied biology mark without enough evidence-based medicals to decide whether to treat using TKIs.

Conventional DNA sequencing, after round pcr expands EGFR gene, though DNA analysis, fluorescence RT-PCR, high performance liquid chromatography EGFR genetic mutation is can determine whether, but is analyzed from technological layer, all there is limitation.1. live body is drawn materials：Belong in vitro, have Invasive inspection, the risk that lesion shifts after increase operation；2. tissue sampling requires high, had a great influence by composition and experiment condition； 3. can the materials that studied at present much from pathologic specimen when making a definite diagnosis, represent patient and receive the preceding recurrence of TKI treatments Neoplastic conditions need to be studied further；4. complex operation, take it is more, randomness is larger；5. expensive equipment, efficiency needed for detection Relatively low, domestic only several units can provide this gene tester at present, therefore be difficult to be widely applied, mesh Preceding only about 1/5th patient can find suitable tissue specimen and be detected；6. newest research is had tried to from thoracic cavity EGFR mutation are detected in hydrops and serum, but the EGFR sample sizes that obtain due to this mode and its small, it is recall rate, consistent Property and accuracy rate are relatively low；7. traditional Imaging Technology in EGFR targeted therapy curative effects are monitored there is also many uncertain factors, The effect of there is no clear and definite iconography means to judge EGFR targeted therapies.Identified in NSCLC patient most possibly from The patient to be benefited in EGFR targeted therapies, it is still the problem for determining EGFR targeted therapies.Therefore, seek a kind of simpler, more straight See, and it is same sensitive, accurate, and sensitive patients can be treated in live body real-time estimate small molecule EGFR inhibitor, instruct such Drug medication, the detection method of monitoring treatment curative effect become another study hotspot problem studied both at home and abroad at present.

(3) molecular image is the ideal style for detecting EGFR states ---¹⁸F marks small molecule, pyrimidine acrylic amide The application of EGFR positive electron tracers

Molecular image is to reach medical diagnosis on disease, efficacy determination on live body from Molecular level study cell function, metabolism With the uncharted field of new drug development.Molecular image utilizes molecular probe combination intracellular certain target molecules, and live body uses PET, PET/ The detection imaging such as CT, MRI, SPECT and optical imaging apparatus, there is high specific, high sensitivity and high-resolution, can be non-invasive Positioning, qualitative and quantitative data are provided.

¹¹C-PD153035 is the PET tracer being imaged earliest for EGFR inhibitor.But due to¹¹C half-life period is only 20 minutes, and 11C-PD153035 has huge uptake in normal liver thus is not as suitable as EGFR PET tracers Agent.Therefore, explore¹⁸The synthetic method of F mark small molecule pyrimidine acrylic amide EGFR class PET tracers just turns into basis With the focus and emphasis of clinical research.From the point of view of the selection of precursor, present medicine be mostly using quinazoline or quinoline amine as The EGFR of basic parent nucleus is reversible or irreversible inhibitor, its poor selectivity to wild-type cell and the toxic side effect brought is also Inevitably.Therefore, urgent need new type, the compound of especially novel skeleton solve drug resistance, selectivity The problems such as poor.

¹⁸The positive electron tracer of F marks is the most frequently used positron emission of clinical PET/CT, PET and coincidence circuit system Property tracer, [¹⁸F]FDG、[¹⁸F]FLT、[¹⁸F] acetate, [¹⁸F] choline etc. as positive electricity radial pattern tracer at present by It is widely used in the early diagnosis of tumour, angiocarpy and the nervous system disease, therapeutic scheme is formulated and curative effect evaluation.But at present On¹⁸F mark third generation EGFR small molecule class positive electron tracers are seldom in clinical practice, particularly clinic¹⁸F is marked just Electronics tracer is mainly using being fluorinated poly- ammonium (kryptofix/K+¹⁸F^-) it is used as phase transfer catalyst (phase transfer Catalysis, PTC), labeling process is relative complex, even with state-of-the-art full-automatic positron radioactive tracer Synthesis system, multistep reaction is also wanted to complete¹⁸F marks small molecule pyrimidine acrylic amide EGFR class positive electron tracers.This Sample not only extends mark time (generally requiring 90-120 minutes), and meeting reduction flag rate, causes to lack with these methods and faces Bed practicality.In addition, scholars couple so far¹⁸The ratio of F mark small molecule pyrimidine acrylic amide EGFR positive electrons tracer researchs It is more.But invertibity EGFR class positive electron tracer holdup times in tumour cell are short, uptake ratio is than relatively low.And can not Inverse property EGFR class positive electron tracers holdup time in tumour cell is long, uptake ratio is also high.So research tumour is taken the photograph with height Take the method for the EGFR class positive electron tracers of the one-step synthesis method of (high-affinity) to turn into molecular image research and be badly in need of what is solved Problem.

The content of the invention

It is an object of the invention to provide one kind¹⁸The pyrimidine acrylic amide EGFR positive electrons tracer and one of F marks Kind can pass through full-automatic, one-step synthesis method¹⁸The pyrimidine acrylic amide EGFR positive electron tracer methods of F marks.This method Synthesized using the precursor of designed, designed of the present invention¹⁸The positron radioactive tracer of F marks, has that multipurpose, yield be big, efficiency Height, method are stable, generated time is short and low cost and other advantages.

Multipurpose, high yield, synthesis stably, efficient and quick and inexpensive are disclosure satisfy that in order to reach to design¹⁸F is marked Remember EGFR positive electrons Tracer Technique and method.The present invention is reached using following key core technological means in the design The technical problem of required solution.These core technologies include：Unique precursor is designed, the present invention is made using pyrimidine acrylamide For the framework compound of labelled precursor, use¹⁸Framework compound and its derivative are marked to complete F.Can from Fig. 1 Go out, on the basis of acrylamide agent structure is constant in holding N- [3- (2- pyrimdinyl-aminos) phenyl], pass through¹⁹F-¹⁸F^-Isotope (IEX) method of displacement is by the organic trifluoroborate of precursor amphion¹⁹F atom is replaced as with active¹⁸F atom；Before Body is using quick and a step mark¹⁹F-¹⁸F^-Isotopic (IEX) method, to improve the mark rate of tracer, specific activity And radiochemicsl purity；Reach the purpose of heavy dose of synthesis and Fast back-projection algorithm using heating；Using HPLC to high specific activity tracer or Splitter is separated to tracer further to shorten product disengaging time；Combined coefficient is improved using microwave heating technique； Full-automatic, one-step synthesis method 18F mark small molecule EGFR class positive electron tracers purpose.

(1) one-step synthesis method¹⁸F marks EGFR positive electron tracers

¹⁹F-¹⁸F^-Isotopic (IEX) method is preferable labeling method.Used in the present invention on precursor¹⁹F-¹⁸F^-Together Plain displacement (IEX) method in position, mild condition and quick mark can improve mark rate, specific activity and radiochemicsl purity.In addition, protecting Organic three fluoboric acid of amphion is introduced on the basis of holding N- [3- (2- pyrimdinyl-aminos) phenyl] acrylamide agent structure being constant Salt groups (raising hydrophily), this is one of core technology of the present invention.

(2) precursor, to improve tracer hydrophily, reduces liver due to introducing the organic trifluoroborate group of amphion Dirty uptake ratio, improve tumor uptake rate.Traditional¹¹The EGFR positive electrons tracer of C flag gathers in liver with largely dense, Its clinical practice is significantly affected.

(3) building-up process is shortened using heating, to meet the mesh of clinical heavy dose of synthesis positron radioactive tracer 's.

(4) utilize HPLC to separation product to obtain high specific activity tracer, or divide using splitter carrying out product Cost is synthesized from reducing.

For needing to separate product after small molecule EGFR class positive electron tracer one-step synthesis, the invention provides use Two methods of HPLC and column isolation technics.The specific activity of product is significantly improved using HPLC, it is obvious using column isolation technics Reduce the cost of synthesis.

(5) combined coefficient is improved using microwave heating technique

Microwave not only has the characteristics of homogeneous heating, be rapidly heated and reduce temperature, and microwave heating significantly improves Combined coefficient.

(6) one-step synthesis method is utilized¹⁸F positive electron tracers

¹⁸The small molecule EGFR class positive electron tracers of F marks, the maximum difficult point of building-up process are final products in precursor base Need to pass through on plinth¹⁹F-¹⁸F^-Isotopic (IEX) method will¹⁸F introduces the organic trifluoroborate group of amphion, is so Raising mark rate conventional method uses 2 steps or multi-step synthetic methods.The special precursor designed using the present invention, well Solve current¹⁸Knotty problem in F mark small molecule EGFR positive electrons tracer synthesis.Building-up process substantially is shortened, is improved Yield.

In order to reach the technology have practicality purpose we in the design especially pay attention to reach full-automatic one-step method The purpose of synthesis.One-step synthesis method can reduce the radiation of operating personnel's ray, reaction condition is gentle, be rapidly completed building-up process, There is high reproducibility and stability.For positron radioactive tracer because the half-life period of nucleic is very short, no Full-automatic one-step synthesis process can be completed, the problem of practicality of just not knowing where to begin.Therefore, the present invention is in GE companies On TracerLab FX fn chemical synthesis devices the whole world first complete one-step synthesis method go out it is above-mentioned¹⁸The positron radioactivity of F marks Tracer.And reach the effect of above-mentioned satisfaction.

Specifically, in order to achieve the above object, present invention employs following technological means：

One kind of the present invention¹⁸The pyrimidine acrylic amide EGFR positive electron tracers of F marks, it is (((2-((2- propylene Acylamino--5- methoxyl groups-4-((4- (1- Methyl-1H-indole-3- bases) pyrimidine -2-base) amino) phenyl) (methyl) amino) second Base) dimethylammonio) methyl) difluoro is (fluoro-¹⁸F) borate, it is named as¹⁸F-BF₃- AZD9291, it has the change shown in Formula II Learn structure：

Further, the invention also provides a kind of using described in full-automatic one-step synthesis method¹⁸The pyrimidine third of F marks The method of acrylamide EGFR positive electron tracers, is carried out successively as follows：

1) accelerator will be come from¹⁸F ion and water pass to receiving bottle and imported into anion-exchange column；

2) using the mixed solution of strong base-weak acid salt and acetonitrile by the anion-exchange column¹⁸F ion is washed Take off and be sent to reaction bulb；

3) precursor is dissolved in DMSO, acetonitrile or DMF solution, adds cushioning liquid to make pH value of solution=2, solution is added to institute State in reaction bulb and heated under the protection of inert gas, the chemical structural formula of the precursor is shown in formula I：

4) temperature is reduced, the crude product in the reaction bulb is purified by SPE C18 pillars；

5) target product to step 4) resulting solution is separated, collected, and is produced.

Wherein, it is preferred that the strong base-weak acid salt is K₂CO₃、KHCO₃、Cs₂CO₃Or one kind in TBAC.

Wherein, it is preferred that the cushioning liquid is pyridazine hydrochloric acid buffer solution.

Wherein, it is preferred that described is heated to be microwave heating or electric heating stove heat.

Wherein, it is preferred that using high performance liquid chromatography or purification column or splitter to target product in step 5)¹⁸F-BF₃- AZD9291 is separated, collected.

Further, the invention also provides described in 1¹⁸The pyrimidine acrylic amide EGFR positive electron tracers of F marks Agent is preparing the application in detecting the horizontal reagent of expression of epidermal growth factor receptor.And

Described¹⁸The pyrimidine acrylic amide EGFR positive electrons tracer of F marks is preparing screening epidermal growth factor receptor Application in body inhibitor reagent.

Utilization proposed by the present invention¹⁸F mark small molecule EGFR class positive electron tracer methods for lung cancer early diagnosis, Lung cancer therapy solution formulation, the evaluation of curative effect are respectively provided with important value.

It is summed up, the present invention has following features compared with prior art：

1st, stably：Utilize¹⁸The small molecule EGFR class positive electrons tracer full-automation of F marks, one-step synthesis method method, are closed Into efficiency high, and stably.This method substantially overcomes that generated time existing for multi-step synthetic methods is long, efficiency is low, is not easy The inherent shortcomings such as grasp.

2nd, routine clinical work is met：Routine clinical production¹⁸F amounts between 1-4Ci, for this just need improve from¹⁸F Post elutes¹⁸F ability, that is, increase strong base-weak acid salt K₂CO₃、KHCO₃、Cs₂CO₃Or the concentration and volume of TBAC elutions are to improve¹⁸F yield, this is the yield of highest synthesis so far.

3rd, it is simple, facilitate building-up process：Synthesis flow is significantly simplify, and establishes a brand-new utilization¹⁸F marks Small molecule EGFR class positive electron tracer class new technological process.

4th, multipurpose：Synthesis¹⁸There is the small molecule EGFR class positive electrons tracer of F marks diagnosis and guiding treatment to face Bed value, it will be played an important role especially for personalized treatment is promoted.

Brief description of the drawings

Fig. 1 is EGFR TKI-BF of the present invention₃The disjunctive path figure of-AZD9291 compounds；

Fig. 2 is BF₃- AZD9291 high performance liquid chromatography (HPLC) result；

Fig. 3 is BF₃- AZD9291 nuclear magnetic spectrum result；

(1)1H-NMR；(2)19F-NMR

Fig. 4 is BF₃- AZD9291 mass spectral results；

Fig. 5 is the process chart of the present invention.

Abbreviation annotation：

EGFR Ep idermal Growth Factor Receptor

TKI T yrosine Kinase Inhibitor

DMF N,N-dimethyl formamide

DMSO Dimethylsulfoxide

IEX Iso tope exchange

Embodiment

The invention will be further described with reference to the accompanying drawings and examples, but the process route of the present invention is not only limited to The content stated below.

Embodiment 1EGFR TKI-BF₃- AZD9291 preparation

Described BF₃- AZD9291 molecular structural formula is shown in formula I:

Preparation flow figure is as shown in figure 1, specific preparation method is as follows：

(1) by 50g (2- chlorine pyrimidine-4-yl) -1- methyl -3a, 7a- dihydro -1H- indoles, 38.03g 4- under nitrogen protection Fluoro- 2- methoxyl groups -5- nitroanilines and 37.09g p-methyl benzenesulfonic acid are mixed in 580ml 2- amylalcohols, and mechanical agitation is anti-at 105 DEG C 2.5h is answered, after being cooled to room temperature, is filtered, filter cake is washed with 2- amylalcohols, drains to obtain light tan solid, carried by column chromatography for separation It is pure, using methanol and dichloromethane according to volume ratio 1:100 eluents being mixed to get, finally give pale yellow needles and consolidate Body is compound one, yield 85%；

(2) it is 65g compounds one, 19.65gN, N, N'- trimethyls ethylenediamine and 26.93N, N- diisopropylethylamine is miscible Stirred in 600mlDMF solvents, 120 DEG C of reaction 4h, solution is poured into frozen water after cooling room temperature and diluted, with 50ml dichloromethanes Alkane extracts three times, merges organic phase and is washed three times with saturated common salt, by organic addition anhydrous sodium sulfate drying, filters, be spin-dried for Purified afterwards by column chromatography for separation, using methanol and dichloromethane according to volume ratio 3:100 eluents being mixed to get, most Rufous needle-like solid, i.e. compound two, yield 62% are obtained eventually；

(3) 3.22g compounds two, 9.62g iron powders and ammonium chloride are added in round-bottomed flask, with the 120mL in flask The mixed solution of ethanol and 30ml water is miscible, heating reflux reaction 3h, and cooling filters, and filtrate concentration is spin-dried for, and passes through SCX posts point From purification, eluent is 0.35M methanolic ammonia solution, obtains brown solid, i.e. compound three, yield 85%；

(4) 1.7g compounds three and 0.73ml diisopropylethylamine are dissolved in 50ml dichloromethane, stirred under ice salt bath The 1ml dichloromethane solutions that 0.34ml acryloyl chlorides are added dropwise are mixed, react 1h, then add 30ml 0.5g sodium hydroxide water at room temperature Solution continues to react 4h.Filter, collect solid and be dried to obtain product, i.e. compound four, yield 39%；

(5) 9.2mg compounds four are placed in 25ml round-bottomed flasks by nitrogen protection at room temperature, add the anhydrous tetrahydrochysene furans of 2ml Mutter dissolving；Another take adds 2ml anhydrous tetrahydro furans and 3 μ l iodomethyl pinacol borate in 25ml round-bottomed flasks, and repeatedly Rinse pipette tips.Iodomethyl pinacol borate tetrahydrofuran solution is slowly added in the solution of compound four in 5min or so, stirred Suction filtration obtains white solid, i.e. compound five, yield 49% after mixing 12h；

(6) compound five is put into round-bottomed flask, sequentially adds 3M, 300 μ l KHF₂The aqueous solution, 4M, 300 μ l HCl The aqueous solution, 200 μ l deionized waters and 600 μ l DMF solutions, react 2h at 45 DEG C, and 10 μ l NH are added after cooling₄The OH aqueous solution Quenching, it is added drop-wise to dropper on small silicagel column with being washed respectively with water and ethyl acetate, collects cleaning solution and extracted with absolute ether, Merge organic phase, with anhydrous sodium sulfate drying, filtering is spin-dried for, and obtains solid product, i.e. compound six, i.e. described epidermal growth Factor receptor tyrosine kinase inhibitor BF₃- AZD9291, yield 85%.

BF₃- AZD9291 high performance liquid chromatography (HPLC) result from HPLC results as shown in Fig. 2 can be seen that this hair The bright BF being prepared₃- AZD9291 pharmaceutical purities are single, free from admixture.BF₃- AZD9291 nuclear magnetic spectrum and mass spectral results point Not as shown in Figure 3 and 4.

The one-step synthesis method of embodiment 2¹⁸F mark pyrimidine acrylic amide EGFR positive electrons tracer (¹⁸F-BF₃- AZD9291)

Flow chart is as shown in figure 5, synthesis step is as follows：

1st, will transmit from medical cyclotron¹⁸F ion and water pass to receiving bottle (1-4Ci), and imported into anion In exchange column QMA；

2nd, 0.15mL K are utilized₂CO₃(0.9mg)+0.5mL acetonitriles elute from QMA posts¹⁸F ion is simultaneously sent to reaction bulb；

3rd, 3mg precursors (embodiment 1 is prepared) are dissolved in 0.5mL 50% (v/v) the DMF aqueous solution, add adjacent phenodiazine Miscellaneous benzene hydrochloric acid buffer solution makes pH value of solution=2, and mixed solution is added into reaction bulb；The water-bath under the protection of nitrogen or helium 80 DEG C, 15 minutes of heating；

4th, temperature is cooled to 25 DEG C, then by reaction solution cross C18 posts separated, purified product, be¹⁸F is marked phonetic Pyridine acrylic amide EGFR positive electron tracers¹⁸F-BF3-AZD9291 (shown in Formula II).

Embodiment 3 uses one-step synthesis method¹⁸F mark pyrimidine acrylic amide EGFR positive electrons tracer (¹⁸F-BF₃- AZD9291)

To be heated in step 3 using 70W microwaves, the heat time can shorten 1-2 minutes, and combined coefficient can reach 30%, Radio-chemical purity is more than 98%, and other steps are same as Example 2.

Experimental example 1BF3-AZD9291 is tested the inhibitory action for being mutated EGFR Lines H1975

1st, test compound：BF3-AZD9291, prepared by the method for embodiment 1

2nd, test method：

Take the logarithm growth period EGFR21 exons point mutation (L858R) merge 20 exon point mutation (T790M) Lines H1975 cells, be inoculated in 10000/hole in 96 orifice plates, in 37 DEG C, 5%CO₂Under the conditions of After culture 24 hours, administration group adds test compound, make its final concentration be respectively 0.01,0.1,1.25,2.5,5,10,20, 40 μM, blank control group adds 0.08% dimethyl sulfoxide (DMSO), after being cultivated 24,48 hours under the conditions of 37 DEG C of 5%CO2, using MTT Colorimetric method, absorbance A value is determined at wavelength 490nm.Half inhibiting rate of the medicine to growth of tumour cell is calculated according to A values (IC50)。

3rd, result of the test

Result of the test shows that the compounds of this invention BF3-AZD9291 has to lung cancer cell line H1975 suppresses its propagation Effect.

Claims (8)

1. It is 1. a kind of¹⁸The pyrimidine acrylic amide EGFR positive electron tracers of F marks, it is characterised in that described EGFR positive electrons show Track agent is (((2-((2- acrylamido-5- methoxyl groups-4-((4- (1- Methyl-1H-indole-3- bases) pyrimidine -2-base) amino) Phenyl) (methyl) amino) ethyl) dimethylammonio) methyl) difluoro (Value linear F) borate, it is named as¹⁸F-BF₃- AZD9291, It has the chemical constitution shown in Formula II：
2. It is 2. a kind of using described in full-automatic one-step synthesis method claim 1¹⁸The pyrimidine acrylic amide EGFR positive electricity of F marks The method of sub- tracer, it is characterised in that carry out successively according to the following steps：

   1) accelerator will be come from¹⁸F ion and water pass to receiving bottle and imported into anion-exchange column；

   2) using the mixed solution of strong base-weak acid salt and acetonitrile by the anion-exchange column¹⁸F ion is eluted simultaneously It is sent to reaction bulb；

   3) precursor is dissolved in DMSO, acetonitrile or DMF solution, adds cushioning liquid to make pH value of solution=2, solution is added to described anti- Answer in bottle, heated under the protection of inert gas, the chemical structural formula of the precursor is shown in formula I：

   4) temperature is reduced, the crude product in the reaction bulb is purified by SPE C18 pillars；

   5) target product to step 4) resulting solution is separated, collected, and is produced.
3. 3. according to the method for claim 2, it is characterised in that：The strong base-weak acid salt is K₂CO₃、KHCO₃、Cs₂CO₃Or One kind in TBAC.
4. 4. according to the method for claim 2, it is characterised in that：The cushioning liquid is that pyridazine hydrochloride buffer is molten Liquid.
5. 5. according to the method for claim 2, it is characterised in that：Described is heated to be microwave heating or electric heating stove heat.
6. 6. according to the method for claim 2, it is characterised in that：High performance liquid chromatography or purification column are utilized in step 5) or is divided From post to target product¹⁸F-BF₃- AZD9291 is separated, collected.
7. 7. described in claim 1¹⁸The pyrimidine acrylic amide EGFR positive electrons tracer of F marks is preparing detection epidermal growth Application in factor acceptor expression reagent.
8. 8. described in claim 1¹⁸The pyrimidine acrylic amide EGFR positive electrons tracer of F marks is preparing screening epidermal growth Application in factor receptor inhibitor reagent.