CN113024565B - Radioactive isotope carbon-14 labeled ibrutinib and synthesis method thereof - Google Patents

Radioactive isotope carbon-14 labeled ibrutinib and synthesis method thereof Download PDF

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CN113024565B
CN113024565B CN202110351880.7A CN202110351880A CN113024565B CN 113024565 B CN113024565 B CN 113024565B CN 202110351880 A CN202110351880 A CN 202110351880A CN 113024565 B CN113024565 B CN 113024565B
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phenoxyphenyl
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ibrutinib
pyrazolo
carbon
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CN113024565A (en
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杨征敏
王国通
李琳港
李书琰
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Zhejiang Aisoto Label Pharmaceutical Technology Co ltd
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Shanghai Qi Zhen Environmental Technology Co ltd
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Abstract

The invention provides a radioactive isotope carbon-14 marked ibrutinib, and preparation method and application thereof, the invention relates to 14 C]Barium carbonate as the starting radioisotope material, warp 14 C]Preparing carbon dioxide, carboxylating, halogenating, condensing, methylating, cyclizing twice, carrying out Mitsunobu reaction, removing protecting groups and the like to obtain carbon-14 labeled ibrutinib, namely pyrazolo [3,4-d ] of ibrutinib molecule for the first time]The preparation of the 3-carbon-14 marker in the pyrimidine fragment provides a viable, economical, and safe method. Furthermore, the carbon-14 labeled ibrutinib can be used as a radioactive tracer, is mainly used for basic research on the environmental safety problem of the drug ibrutinib, and can also be used for basic research on the radioactive pharmacokinetics of tissue distribution, metabolite structure identification, mass balance and the like of the drug. Meanwhile, the method of the invention provides reference for the preparation of the carbon-14-containing synthetic building block.

Description

Radioactive isotope carbon-14 labeled ibrutinib and synthesis method thereof
Technical Field
The invention belongs to the field of radiochemical synthesis, and particularly relates to radioisotope carbon-14 labeled ibrutinib ((R) -1- (3- (4-amino-3- (4-phenoxyphenyl) -1H- [ 3-) 14 C]Pyrazolo [3,4-d]Pyrimidin-1-yl) piperidin-1-yl) prop-2-en-1-one).
Background
Currently, medicine is one of the important living materials necessary for human health care and disease treatment. For a long time, people often rely on medicines for treating self health-care diseases and diseases, so that the medicines are largely used in daily life of people for a long time. It has been found that many pharmaceutical residues are detected in wastewater and sludge (including biomass entrapped therein) discharged from many municipal sewage pipes around the world. In order to save water resources and protect the environment, the waste water is often recycled by people (such as irrigation), and polluted sludge can be returned to the natural environment after specialized treatment. However, in the process of recycling and reusing wastewater and treating polluted sludge, the drug residues may reenter the pollution-free water source and soil, and then be absorbed by crops, livestock, fish, shrimp, crab and other animals required by people's diet, and finally enter human bodies through food chains, and the drug residues entering human bodiesThe retentate and its metabolic degradants present potential safety risks to human health. Thus, researchers at home and abroad are increasingly concerned about The scientific problems of The origin, metabolic mechanism, degradation rule, environmental behavior and tendency of drug residues in waste water and sludge discharged from municipal sewage pipes, which are closely related to human health and safety [ The occucure of pharmaceuticals, personal care products, endo-drugs and scientific drugs in surface water in South waters, uk. water Res,2008,42, 3498-3518; biological interaction of pharmaceuticals and other anthropogenic waste indicators in earth words from acquired soil tissue with biosolid or thread manure, environ Sci Technol,2008,42, 1863-1870; environ polar, 2013,182,150-156; 312-316 in ambient strain of pharmaceutical products for human use in the localized procedure. Regul Toxicol Pharmacol,2014,68 (3); degradation and transformation products of acetaminophen in water Res,2014,49, 44-52; phototranformation and metabolic pathways of 14 C-carbamazepine in carrot and celery.J Agric Food Chem,2020,68(11):3362～3371；Pharmaceuticals and personal care products in archived U.S.biosolids from the 2001 EPA national sewage sludge survey.Water Res,2010,44,658～668；Environmental fate of pharmaceuticals in water/sediment systems.Environ Sci Technol,2005,39,5209～5218]. With the continuous and rapid advance of urbanization in China and the continuous improvement of the living standard and quality of people, the amount of wastewater and sludge discharged from municipal sewage pipelines is increased sharply, and the research on the environmental safety related to medicines is more and more important and urgent.
Ibrutinib (CAS number: 936563-96-1; English name: Ibrutinib; Chinese name: (R) -1- (3- (4-amino-3- (4-phenoxyphenyl) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl) piperidin-1-yl) prop-2-en-1-one) is a Bruton's Tyrosine Kinase (BTK) inhibitor for the treatment of Chronic Lymphocytic Leukemia (CLL), Mantle Cell Lymphoma (MCL) and Fahrenheit giant globulinemia (WM). The common chemoimmunotherapy has no target to MCL and CLL, and the cancer has difficult cure and easy recurrence, but ibrutinib can combine with BTK target to irreversibly inhibit the activity of BTK, thereby influencing the normal metabolism of B lymphocyte to effectively inhibit the proliferation and survival of tumor cells. By 2013, ibrutinib was approved for marketing in the united states; 10 months 2014, ibrutinib was approved for the market in the european union; in 2015, ibrutinib was approved for sale in japan. At present, China approves the import of the medicine, and simultaneously, various medicine enterprises in China are imitating the medicine to prepare for the production of subsequent raw medicines and capsules. So far, the researches on the problems of the metabolic mechanism, the degradation rule, the environmental behavior, the tendency and the potential safety risk to the environment of the medicine in the environment are rarely reported in a public way.
The radioactive isotope carbon-14 labeled ibrutinib is a tracer agent which is necessary for basic research on environmental safety problems and the like of the medicine by means of radioactive isotope labeling technology. In the carbon-14 labeled synthesis of ibrutinib, sites with chemical stability and metabolic stability in the molecule are often selected to ensure that labeled atoms are not dropped off as much as possible during the tracer test. The literature reports an ibrutinib carbon-14 marker for radioactive pharmacokinetic study, the marker site of which is carbonyl carbon in molecule [ adsorption, metabolism, and interaction of oral 14 C radiolabeled Ibrutinib:an open-label,phase I,single-dose study in healthy men.Drug Metab Dispos,2015,43(2):289～297]See fig. 3. Although the marker has short synthesis steps and low cost, the synthesis of the marker must use a low-boiling point intermediate [ carbonyl- 14 C]Acryloyl chloride, this intermediate, is very volatile during experimental operations, and is very likely to be inhaled into the body by experimenters inadvertently 14 C, internal irradiation harms the health; in addition, the trace amount of the substance is also easily deteriorated. More importantly, the carbon-14 labeled acryloyl fragment in the marker reported in the literature can not be tracked due to the easy detachment from the molecular skeleton, which causes the motion change trace of the main fragment of the molecule, and leads to incomplete and even paradoxical information obtained by the isotope labeling test.
Pyrazolo [3,4-d]The pyrimidine fragment is positioned at the core part of the ibrutinib moleculeAromatic, and is the preferred carbon-14 labeling unit. Therefore, any site containing carbon atoms in the labeling unit can be selected for labeling (synthesis and analysis of the radioactive isotope carbon-14 labeled chlorpyrifos. isotope 2020,33(6): 409-416), and the carbon-14 labels of the sites are firm and are not easy to fall off. From the viewpoint of the synthesis technical route of cold reaction, the method adopts 14 C]Formamide is used as the starting radioisotope to prepare 6-carbon-14 labeled ibrutinib [ (R) -1- (3- (4-amino-3- (4-phenoxyphenyl) -1H- [6- 14 C]Pyrazolo [3,4-d]Pyrimidin-1-yl) piperidin-1-yl) prop-2-en-1-one as shown in FIG. 2]And the requirements of downstream tracing tests can be met. However, a large excess of [2 ] must be used in the thermal reaction for preparing the marker 14 C]Formamide, high cost of label synthesis, and 14 C]the preparation process of formamide involves volatile 14 C]Formic acid, which has a cause for the operator 14 Potential risk of illumination within C.
In view of the above, the invention selects pyrazolo [3,4-d ] pyrimidine fragment in ibrutinib molecule as a labeling unit, and carbon-14 labeling is carried out by using 3-position in the labeling unit as a labeling site. The carbon-14 mark of the site is firm and is not easy to fall off, the cost of mark synthesis is low, and the operation is safe; the obtained carbon-14 marker can be used as a radioactive tracer, and not only can be used for researching the environmental safety problem of the medicine, but also can be used for radioactive pharmacokinetics research of tissue distribution, metabolite structure identification, mass balance and the like of the medicine.
Disclosure of Invention
In view of the above, the present application provides a carbon-14 labeled ibrutinib [ (R) -1- (3- (4-amino-3- (4-phenoxyphenyl) -1H- [3- 14 C]Pyrazolo [3,4-d]Pyrimidin-1-yl) piperidin-1-yl) prop-2-en-1-one; hereinafter, carbon-14 labeled ibrutinib) and a synthetic method thereof, which provide a necessary tracer for developing basic research works such as environmental safety problems and radioactive pharmacokinetics of ibrutinib, and the like, and the structural formula of the tracer is as follows:
Figure BDA0003002369060000031
the invention has the beneficial effects that:
the marker position in the marker is positioned at the core part of an ibrutinib molecular skeleton, and the marker locus is positioned in a pyrazolo [3,4-d ] pyrimidine marker unit with aromaticity (the aromatic chemical structure is more stable), so that the carbon-14 marker is firm, and the marker can reveal the movement change information of ibrutinib in a complex system in a tracing test.
The invention also provides a synthetic method of the radioisotope carbon-14 labeled ibrutinib, which is characterized by comprising the following steps:
s1: under the protection of inert gas and at the low temperature of < -70 ℃, 4-phenoxyphenyl lithium (1) is reacted with sodium benzoate 14 C]Barium carbonate released [ alpha ], [ alpha ] and [ alpha ], [ alpha ] barium carbonate 14 C]CO 2 Reaction preparation of 4-phenoxybenzene [ alpha ], [ beta ] -pheno ] or a mixture thereof 14 C]Formic acid (2);
s2: under the protection of inert gas and at the reaction temperature of 0-80 ℃, 4-phenoxybenzene [ alpha ], [ alpha ] an 14 C]The formic acid (2) reacts with thionyl chloride or oxalyl chloride to produce 4-phenoxybenzene [2 ] 14 C]Formyl chloride; reacting the obtained acyl chloride with sodium hydrogen and malononitrile in tetrahydrofuran for 3-15 h, and performing conventional post-treatment and column chromatography purification to obtain 2- (hydroxy- (4-phenoxyphenyl) (, [2 ]) 14 C]Methylene) malononitrile (3);
s3: under the protection of inert gas and at the reaction temperature of 0-30 ℃, the intermediate 2- (hydroxy- (4-phenoxyphenyl) (+) 14 C]Methylene) malononitrile (3), trimethylsilyldiazomethane and organic alkali are stirred in acetonitrile for 12-48 h, and the reaction is finished; adjusting the reaction solution to weak acidity (pH about 6) with dilute hydrochloric acid, and purifying by conventional post-treatment and column chromatography to obtain 2- (methoxy- (4-phenoxyphenyl), [2 ] 14 C]Methylene) malononitrile (4);
s4: under the protection of inert gas and at the reaction temperature of 50-100 ℃, the intermediate 2- (methoxy- (4-phenoxyphenyl) [ ] 14 C]Methylene) malononitrile (4) and hydrazine hydrate are stirred in lower alcohol for 1-3 h; obtaining 5-amino-3- (4-phenoxyphenyl) -1H- [3- 14 C]Pyrazole-4-carbonitrile (5);
s5: under the protection of inert gas and at a reaction temperature of 80-200 ℃, the intermediate 5-amino-3- (4-phenoxyphenyl) -1H- [3- 14 C]Pyrazole-4-cyanide (5) is reacted with formamide; after the reaction is finished, the reaction mixture is treated by conventional method to obtain the intermediate 3- (4-phenoxyphenyl) -1H- [3- 14 C]Pyrazolo [3,4-d]Pyrimidin-4-amine (6);
s6: under the protection of inert gas and low temperature reaction at-5 to 30 ℃, the intermediate 3- (4-phenoxyphenyl) -1H- [3- 14 C]Pyrazolo [3,4-d]Reacting pyrimidin-4-amine (6) with (S) -1-tert-butoxycarbonyl-3-hydroxypiperidine, triphenylphosphine and azodicarboxylic acid diiso in tetrahydrofuran; after the reaction is finished, (R) -3- (4-amino-3- (4-phenoxyphenyl) -1H- [3- 14 C]Pyrazolo [3,4-d]Pyrimidin-1-yl) piperidine-1-carboxylic acid tert-butyl ester (7);
s7: at a reaction temperature of 10-40 ℃, the intermediate (R) -3- (4-amino-3- (4-phenoxyphenyl) -1H- [3- 14 C]Pyrazolo [3,4-d]Removing a protecting group from tert-butyl pyrimidin-1-yl) piperidine-1-carboxylate (7) in an acidic tetrahydrofuran solution or a dichloromethane solution; after the reaction is finished, (R) -3- (4-phenoxyphenyl) -1- (piperidine-3-yl) -1H- [3- 14 C]Pyrazolo [3,4-d]Pyrimidin-4-amine (8);
s8: under the protection of inert gas and at the reaction temperature of 0-30 ℃, the intermediate (R) -3- (4-phenoxyphenyl) -1- (piperidin-3-yl) -1H- [3- 14 C]Pyrazolo [3,4-d]Reacting pyrimidine-4-amine (8), triethylamine and allyl chloride in dichloromethane; after the reaction is finished, (R) -1- (3- (4-amino-3- (4-phenoxyphenyl) -1H- [ 3-) 14 C]Pyrazolo [3,4-d]Pyrimidin-1-yl) piperidin-1-yl) prop-2-en-1-one (9).
The synthesis method has the beneficial effects that:
the starting radioisotope raw material used in the labeling synthesis of the present invention is [2 ] 14 C]The barium carbonate has low raw material cost; during operation, the radioactive gas is introduced 14 C]CO 2 The preparation and the subsequent carboxylation reaction are carried out in a strictly sealed system and cannot escape,while other radioisotopic carbon-14-containing intermediates are viscous pastes or non-sublimable solids which do not cause operator problems 14 C internal irradiation, so the operation safety is high.
Further, the inert gas is argon or nitrogen.
Further, in S1, 4-phenoxybenzene [2 ] 14 C]The formic acid (2) can also be reacted with the Grignard reagent 4-phenoxyphenylmagnesium halide under protection of an inert gas at room temperature 14 C]CO 2 Preparation by reaction (2), in the reaction 14 C]CO 2 When the consumption is complete, the reaction is finished, and the 4-phenoxybenzene can be obtained by conventional post-treatment 14 C]Formic acid (2).
Further, in S2, the halogen atom of the Grignard reagent 4-phenoxyphenylmagnesium halide is Cl, Br, I.
Further, in S3, the organic base is selected from one or both of diisopropylethylamine and triethylamine.
Further, in S4, the lower alcohol is selected from one or more of methanol, ethanol, and propanol.
The invention also provides a carbon-14-containing synthetic building block synthesized by the synthesis method.
The invention also provides a radioisotope carbon-14 labeled ibrutinib, the marker can be used as a radioactive tracer, is mainly used for basic research on the environmental safety problem of the drug ibrutinib, can also be used for basic research on the radioactive pharmacokinetics of tissue distribution, metabolite structure identification, mass balance and the like of the drug, and the carbon-14 is not easy to fall off in the tracing process and is more stable.
Drawings
FIG. 1 shows the chemical structure of carbon-14 labeled ibrutinib (3-carbon-14 label) according to the present invention.
FIG. 2 shows the chemical structure of the 6-carbon-14 marker in the ibrutinib molecule.
FIG. 3 shows the chemical structure of carbonyl carbon-14 marker in ibrutinib molecule.
FIG. 4 is a synthetic route of carbon-14 labeled ibrutinib of the present invention.
Fig. 5 is a nuclear magnetic resonance hydrogen spectrum of the radioactive intermediate (2) of the present invention.
Fig. 6 is a nuclear magnetic resonance hydrogen spectrum of the radioactive intermediate (3) of the present invention.
Fig. 7 is a nuclear magnetic resonance hydrogen spectrum of the radioactive intermediate (4) of the present invention.
Fig. 8 is a nuclear magnetic resonance hydrogen spectrum of the radioactive intermediate (5) of the present invention.
Fig. 9 is a nuclear magnetic resonance hydrogen spectrum of the radioactive intermediate (6) of the present invention.
Fig. 10 is a nuclear magnetic resonance hydrogen spectrum of the radioactive intermediate (7) of the present invention.
Fig. 11 is a nuclear magnetic resonance hydrogen spectrum of the radioactive intermediate (8) of the present invention.
FIG. 12 shows the NMR spectrum of carbon-14 labeled ibrutinib (9) according to the present invention.
Detailed Description
The embodiments of the present invention are described below with reference to specific embodiments, and other advantages and effects of the present invention will be easily understood by those skilled in the art from the disclosure of the present specification. The invention is capable of other and different embodiments and of being practiced or of being carried out in various ways, and its several details are capable of modification in various respects, all without departing from the spirit and scope of the present invention.
In addition, FIG. 1 shows the chemical structure of carbon-14 labeled ibrutinib (3-carbon-14 label) according to the present invention; FIG. 2 shows the chemical structure of the 6-carbon-14 marker in an ibrutinib molecule; FIG. 3 shows the chemical structure of carbonyl carbon-14 marker in ibrutinib molecule; FIG. 4 is a synthetic route of carbon-14 labeled ibrutinib of the present invention; FIG. 5 is a nuclear magnetic resonance hydrogen spectrum of the radioactive intermediate (2) of the present invention; FIG. 6 is a nuclear magnetic resonance hydrogen spectrum of the radioactive intermediate (3) of the present invention; FIG. 7 is a NMR spectrum of the radioactive intermediate (4) of the present invention; FIG. 8 is a nuclear magnetic resonance hydrogen spectrum of the radioactive intermediate (5) of the present invention; FIG. 9 is a nuclear magnetic resonance hydrogen spectrum of the radioactive intermediate (6) of the present invention; FIG. 10 is a nuclear magnetic resonance hydrogen spectrum of the radioactive intermediate (7) of the present invention; FIG. 11 is a nuclear magnetic resonance hydrogen spectrum of the radioactive intermediate (8) of the present invention; FIG. 12 shows the NMR spectrum of carbon-14 labeled ibrutinib (9) according to the present invention.
Example 1:
s1: under the protection of argon gas and at-78 deg.C, concentrated sulfuric acid (5mL,18.4M) was slowly added dropwise to the radioactive raw material [2 ] 14 C]Barium carbonate (28.8. mu. Ci,0.5mmol), escape [ alpha ], [ alpha ] and [ alpha ], [ alpha ] and [ alpha ], [ alpha ] or [ alpha ] and [ alpha ], [ alpha ] and [ alpha ], [ alpha ] and [ alpha ] 14 C]CO 2 A solution of 4-phenoxyphenyllithium (0.55mmol) as a metal reagent in tetrahydrofuran (5mL) was introduced through a thin glass tube. Stirring for 30min, reacting, slowly heating to room temperature, cooling in ice-water bath, adding water (5mL) to quench reaction, extracting water phase with dichloromethane, adjusting pH of residual water phase to 4 with dilute hydrochloric acid (1M), extracting with dichloromethane (4mL) for 5 times, mixing organic phases, drying with anhydrous sodium sulfate, vacuum filtering, concentrating under reduced pressure, and drying to obtain 4-phenoxybenzene [2 ] 14 C]Formic acid (2, 25.9. mu. Ci,96mg, radiochemical yield 90%). 1 H NMR(400MHz,DMSO-d 6 )δ:12.82(s,1H),7.94(d,J＝8.7Hz,2H),7.46(t,J＝7.8Hz,2H),7.24(t,J＝7.4Hz,1H),7.15–7.09(m,2H),7.02(d,J＝8.7Hz,2H).ESI-MS m/z:214[M-H] - ,215[M+2-H] - 。
S2: under the protection of argon and at 0 ℃, 4-phenoxybenzene [2 ] 14 C]Adding formic acid (2,20.0 mu Ci) into a dry Schlenk reaction tube, adding thionyl chloride (2mL), placing the reaction tube in an oil bath, heating to 80 ℃, stirring for 3 hours, and removing excessive thionyl chloride by reduced pressure evaporation; the preparation acid chloride, sodium hydride (0.69mmol) was dissolved in tetrahydrofuran (4mL) under argon and 0 deg.C, a solution of malononitrile (0.43mmol) in tetrahydrofuran (2mL) was added dropwise and stirred at room temperature for 15 h. Radio-TLC analysis shows that after radioactive raw material is completely consumed, water (5mL) is added for quenching, diluted hydrochloric acid is added for adjusting to be neutral, ethyl acetate (4mL) is extracted for 5 times, the ethyl acetate phase is dried by anhydrous sodium sulfate, is filtered by suction, and is subjected to reduced pressure concentration column chromatography (ethyl acetate (V): petroleum ether (V): 1:4) to obtain 2- (hydroxy- (4-phenoxyphenyl) [2- 14 C]Methylene) malononitrile (3, 14.0. mu. Ci,52mg, radiochemical yield 70%). 1 H NMR(400MHz,DMSO-d 6 )δ:7.66–7.60(m,2H),7.46–7.39(m,2H),7.18(t,J＝7.4Hz,1H),7.10–7.04(m,2H),6.95(dd,J＝8.5,6.6Hz,2H).ESI-MS m/z:261[M-H] - ,263[M+2-H] - 。
S3: under argonUnder protection and cooling in an ice-water bath (0 ℃), 2- (hydroxy- (4-phenoxyphenyl) [2- 14 C]Methylene) malononitrile (3, 15.0. mu. Ci), diisopropylethylamine (0.40mmol) in acetonitrile (2mL), 1M trimethylsilyldiazomethane in tetrahydrofuran (0.40mL) was added dropwise and stirred at room temperature for 48 h; Radio-TLC analysis shows that after radioactive raw material is completely consumed, water (5mL) is added for quenching, diluted hydrochloric acid is added for adjusting the pH value to 6, ethyl acetate (5mL) is added for extraction for 3 times, liquid separation is carried out, the ethyl acetate phase is dried by anhydrous sodium sulfate, suction filtration is carried out, and reduced pressure concentration column chromatography is carried out (ethyl acetate (V): petroleum ether (V): 1:4) to obtain 2- (methoxy- (4-phenoxyphenyl) [ 2-) 14 C]Methylene) malononitrile (4, 12.0. mu. Ci,57mg, radiochemical yield 80%). 1 H NMR(400MHz,DMSO-d 6 )δ:7.71(d,J＝8.6Hz,2H),7.49(t,J＝7.9Hz,2H),7.28(t,J＝7.4Hz,1H),7.16(t,J＝9.0Hz,4H),3.93(s,3H).ESI-MS m/z:277[M+H] + ,279[M+2+H] + 。
S4: under the protection of argon, 2- (methoxy- (4-phenoxyphenyl) [2- 14 C]Methylene) malononitrile (4,12.0 mu Ci) and hydrazine hydrate (0.446mmol) are dissolved in methanol (4mL), placed in an oil bath, heated to 50 ℃ and stirred for 3 h; TLC (thin layer chromatography) is used for monitoring the completion of the reaction of the radioactive raw material, water (1mL) is added for quenching, the solvent is removed by rotary removal under reduced pressure, ethyl acetate (5mL) is extracted for 3 times, the ethyl acetate phase is washed by saturated ammonium chloride solution (2mL) and saturated sodium bicarbonate solution (2mL) in turn, dried by anhydrous sodium sulfate, filtered by suction, and subjected to column chromatography by concentration under reduced pressure (ethyl acetate (V): petroleum ether (V) ═ 1:1) to obtain 5-amino-3- (4-phenoxyphenyl) -1H- [3- 14 C]Pyrazole-4-carbonitrile (5, 10.2. mu. Ci,54mg, radiochemical yield 85%). 1 H NMR(400MHz,DMSO-d 6 )δ:12.11(s,1H),7.80(d,J＝8.6Hz,2H),7.43(dd,J＝17.3,9.4Hz,2H),7.17(t,J＝7.4Hz,1H),7.14-7.06(m,4H),6.46(s,2H).ESI-MS m/z:277[M+H] + ,279[M+2+H] + 。
S5: under the protection of argon, 5-amino-3- (4-phenoxyphenyl) -1H- [3- 14 C]Dissolving pyrazole-4-cyanogen (5,12.0 mu Ci) in formamide (4mL), placing in an oil bath, heating to 80 ℃, and stirring for 15 h; TLC monitoring radioactive starting material reaction completion, cooling to room temperature, adding water (10mL) to quench, extracting with ethyl acetate (4mL) for 3 times, washing ethyl acetate phase with saturated ammonium chloride solution (4mL) and saturated sodium bicarbonate solution (4mL) in sequenceWashing, drying with anhydrous sodium sulfate, vacuum filtering, concentrating under reduced pressure, and performing column chromatography (ethyl acetate (V): petroleum ether (V) ═ 4:1) to obtain 3- (4-phenoxyphenyl) -1H- [3- 14 C]Pyrazolo [3,4-d]Pyrimidin-4-amine (6, 6.4. mu. Ci,34mg, radiochemical yield 53%). 1 H NMR(400MHz,DMSO-d 6 )δ:13.54(s,1H),8.22(s,1H),7.67(d,J＝8.6Hz,2H),7.43(t,J＝7.9Hz,2H),7.22–7.08(m,5H).ESI-MS m/z:304[M+H] + ,306[M+2+H] + 。
S6: under the protection of argon and at 0 ℃, 3- (4-phenoxyphenyl) -1H- [3- 14 C]Pyrazolo [3,4-d]Pyrimidin-4-amine (6, 10.0. mu. Ci), (S) -1-tert-butoxycarbonyl-3-hydroxypiperidine (0.55mmol), triphenylphosphine (0.55mmol) and tetrahydrofuran (4mL) dissolved in tetrahydrofuran, diisopropyl azodicarboxylate (0.55mmol) was added dropwise in tetrahydrofuran (1mL) under ice-bath, and the mixture was stirred at room temperature for 1 hour; after the radioactive raw materials completely react, adding water (5mL) for quenching, adding saturated sodium bicarbonate to adjust the pH value to 8-9, extracting with dichloromethane (5mL) for 3 times, drying with anhydrous sodium sulfate, performing suction filtration, performing reduced pressure concentration, and purifying by column chromatography (ethyl acetate) to obtain (R) -3- (4-amino-3- (4-phenoxyphenyl) -1H- [3- 14 C]Pyrazolo [3,4-d]Pyrimidin-1-yl) piperidine-1-carboxylic acid tert-butyl ester (7, 6.5. mu. Ci,55mg, radiochemical yield 65%). 1 H NMR(400MHz,DMSO-d 6 )δ:8.25(s,1H),7.66(d,J＝8.4Hz,2H),7.44(t,J＝8.0Hz,2H),7.18–7.12(m,3H),7.08(d,J＝8.0Hz,2H),4.70–4.6(m,1H),4.01–3.5(m,3H),4.12–4.06(m,1H),2.26–2.16(m,1H),2.09–2.03(m,1H),1.61–1.50(m,1H),1.49–1.22(m,9H).ESI-MS m/z:487[M+H] + ,489[M+2+H] + 。
S7: under the protection of argon and at 0 ℃, reacting (R) -3- (4-amino-3- (4-phenoxyphenyl) -1H- [3- 14 C]Pyrazolo [3,4-d]Dissolving tert-butyl pyrimidin-1-yl) piperidine-1-carboxylate (7, 6.0. mu. Ci) in tetrahydrofuran (4mL), dropwise adding concentrated hydrochloric acid (0.5mL), placing in an oil bath, heating to 10 ℃, and stirring for 3 h; after the radioactive raw materials completely react, adding saturated sodium bicarbonate to adjust the pH value to 8-9, extracting with dichloromethane (5mL) for 3 times, drying with anhydrous sodium sulfate, filtering, decompressing and concentrating to obtain (R) -3- (4-phenoxyphenyl) -1- (piperidine-3-yl) -1H- [3- 14 C]Pyrazolo [3,4-d]Pyrimidin-4-amine (8, 4.9. mu. Ci,33mg, radiochemical yield 82%). 1 H NMR(400MHz,DMSO-d 6 )δ:8.23(s,1H),7.65(d,J＝8.6Hz,2H),7.42(t,J＝8.0Hz,2H),7.21–7.08(m,5H),4.75–4.58(m,1H),3.07(d,J＝9.1Hz,1H),2.92(dd,J＝21.4,10.2Hz,2H),2.46(d,J＝12.0Hz,1H),2.16–2.00(m,2H),1.74(d,J＝12.8Hz,1H),1.55(dd,J＝25.0,12.4Hz,1H).ESI-MS m/z:387[M+H] + ,389[M+2+H] + 。
S8: under the protection of argon and at 0 ℃, reacting (R) -3- (4-phenoxyphenyl) -1- (piperidin-3-yl) -1H- [3- 14 C]Pyrazolo [3,4-d]Pyrimidin-4-amine (8, 6.0. mu. Ci) and triethylamine (0.26mmol) were dissolved in dichloromethane (4mL), allyl chloride (0.13mmol) was added dropwise, the temperature was raised to 30 ℃ and stirring was carried out for 30 min. After the radioactive raw material completely reacts, water (4mL) is added for quenching, dichloromethane (5mL) is used for extraction for 3 times, saturated ammonium chloride is used for washing (3mL), anhydrous sodium sulfate is used for drying, suction filtration and reduced pressure concentration are carried out, and (R) -1- (3- (4-amino-3- (4-phenoxyphenyl) -1H- [3- 14 C]Pyrazolo [3,4-d]Pyrimidin-1-yl) piperidin-1-yl) prop-2-en-1-one (9, 5.4. mu. Ci,41mg, radiochemical yield 90%). Analysis by conventional methods (see: methods: novel herbicide propyribac-propyl ester A Ring) 14 The chemical report, 2005,63(21): 1999-2003) shows that the chemical purity of the target carbon-14 labeled ibrutinib (9) is more than 98%, and the radiochemical purity is more than 99%. 1 H NMR(400MHz,DMSO-d 6 )δ:8.26(s,2H),7.66(d,J＝7.3Hz,2H),7.43(t,J＝7.9Hz,2H),7.22–7.09(m,5H),6.89–6.83(m,0.5H),6.76–6.66(m,0.5H),6.10(dd,J＝26.4,16.7Hz,2H),5.71(d,J＝10.8Hz,0.5H),5.59(d,J＝10.2Hz,0.5H),4.79–4.63(m,1H),4.59–4.50(m,0.5H),4.26–4.14(s,1H),4.05(d,J＝10.9Hz,0.5H),3.74–3.66(m,0.5H),3.25-3.14(m,1H),3.05–2.95(s,0.5H),2.33-2.19(m,1H),2.18–2.06(m,1H),1.92–1.89(m,1H),1.66–1.51(m,1H).ESI-MS m/z:441[M+H] + ,443[M+2+H] + 。
Example 2:
s1: under the protection of argon gas and at room temperature, concentrated sulfuric acid (5mL,18.4M) was slowly dropped into the radioactive raw material [2 ] 14 C]In barium carbonate (28.8. mu. Ci,0.5mmol), escape [2 ] 14 C]CO 2 A tetrahydrofuran solution (5mL) of a metal reagent, 4-phenoxyphenylmagnesium iodide (0.55mmol), was introduced through a thin glass tube. After stirring for 30min, the reaction was completed and the system was slowly raised to roomAdding water (5mL) under cooling in ice-water bath to quench reaction, extracting water phase with dichloromethane, adjusting pH of residual water phase to 4 with dilute hydrochloric acid (1M), extracting with dichloromethane (4mL) for 5 times, mixing organic phases, drying with anhydrous sodium sulfate, vacuum filtering, concentrating under reduced pressure, and drying to obtain 4-phenoxybenzene [2 ] 14 C]Formic acid (2, 24.7. mu. Ci, radiochemical yield 86%). 1 H NMR(400MHz,DMSO-d 6 )δ:12.82(s,1H),7.94(d,J＝8.7Hz,2H),7.46(t,J＝7.8Hz,2H),7.24(t,J＝7.4Hz,1H),7.15–7.09(m,2H),7.02(d,J＝8.7Hz,2H).ESI-MS m/z:214[M-H] - ,215[M+2-H] - 。
S2: under the protection of argon and at 0 ℃, 4-phenoxybenzene [2 ] 14 C]Adding formic acid (2,20.0 mu Ci) into a dry Schlenk reaction tube, adding thionyl chloride (2mL), placing the reaction tube in an oil bath, heating to 40 ℃, stirring for 2 hours, and removing excess thionyl chloride by reduced pressure evaporation; the preparation of the acid chloride, sodium hydride (0.69mmol) was dissolved in tetrahydrofuran (4mL) under argon and 20 ℃ and a solution of malononitrile (0.43mmol) in tetrahydrofuran (2mL) was added dropwise and stirred for 4.5 h. Radio-TLC analysis shows that after radioactive raw material is completely consumed, water (5mL) is added for quenching, diluted hydrochloric acid is added for adjusting to be neutral, ethyl acetate (4mL) is extracted for 5 times, the ethyl acetate phase is dried by anhydrous sodium sulfate, is filtered by suction, and is subjected to reduced pressure concentration column chromatography (ethyl acetate (V): petroleum ether (V): 1:4) to obtain 2- (hydroxy- (4-phenoxyphenyl) [2- 14 C]Methylene) malononitrile (3, 14.8. mu. Ci, radiochemical yield 74%). 1 H NMR(400MHz,DMSO-d 6 )δ:7.66–7.60(m,2H),7.46–7.39(m,2H),7.18(t,J＝7.4Hz,1H),7.10–7.04(m,2H),6.95(dd,J＝8.5,6.6Hz,2H).ESI-MS m/z:261[M-H] - ,263[M+2-H] - 。
S3: 2- (hydroxy- (4-phenoxyphenyl) [2- 14 C]Methylene) malononitrile (3, 15.0. mu. Ci) and triethylamine (0.40mmol) were dissolved in acetonitrile (2mL), 1M trimethylsilyl diazomethane in tetrahydrofuran (0.40mL) was added dropwise, and the mixture was stirred at room temperature for 17 hours; Radio-TLC analysis shows that after radioactive material is completely consumed, water (5mL) is added for quenching, diluted hydrochloric acid is added for adjusting the pH value to 6, ethyl acetate (5mL) is added for extraction for 3 times, liquid separation is carried out, the ethyl acetate phase is dried by anhydrous sodium sulfate, suction filtration is carried out, reduced pressure concentration and column chromatography are carried out (ethyl acetate (V): petroleum ether (V): 1:4) to obtain 2- (methoxy- (4-benzene)Oxyphenyl) [2- 14 C]Methylene) malononitrile (4, 11.6. mu. Ci, radiochemical yield 77%). 1 H NMR(400MHz,DMSO-d 6 )δ:7.71(d,J＝8.6Hz,2H),7.49(t,J＝7.9Hz,2H),7.28(t,J＝7.4Hz,1H),7.16(t,J＝9.0Hz,4H),3.93(s,3H).ESI-MS m/z:277[M+H]+,279[M+2+H]+。
S4: under the protection of argon, 2- (methoxy- (4-phenoxyphenyl) [2- 14 C]Methylene) malononitrile (4,12.0 mu Ci) and hydrazine hydrate (0.446mmol) are dissolved in ethanol (4mL), and the solution is placed in an oil bath to be heated to 80 ℃ and stirred for 2 h; TLC (thin layer chromatography) is used for monitoring the completion of the reaction of the radioactive raw material, water (1mL) is added for quenching, the solvent is removed by rotary removal under reduced pressure, ethyl acetate (5mL) is extracted for 3 times, the ethyl acetate phase is washed by saturated ammonium chloride solution (2mL) and saturated sodium bicarbonate solution (2mL) in turn, dried by anhydrous sodium sulfate, filtered by suction, and subjected to column chromatography by concentration under reduced pressure (ethyl acetate (V): petroleum ether (V) ═ 1:1) to obtain 5-amino-3- (4-phenoxyphenyl) -1H- [3- 14 C]Pyrazole-4-carbonitrile (5, 7.9. mu. Ci, radiochemical yield 66%). 1 H NMR(400MHz,DMSO-d 6 )δ:12.11(s,1H),7.80(d,J＝8.6Hz,2H),7.43(dd,J＝17.3,9.4Hz,2H),7.17(t,J＝7.4Hz,1H),7.14-7.06(m,4H),6.46(s,2H).ESI-MS m/z:277[M+H] + ,279[M+2+H] + 。
S5: under the protection of argon, 5-amino-3- (4-phenoxyphenyl) -1H- [3- 14 C]Dissolving pyrazole-4-cyanogen (5,12.0 mu Ci) in formamide (4mL), placing in an oil bath, heating to 140 ℃, and stirring for 6 h; TLC (thin layer chromatography) is used for monitoring the complete reaction of the radioactive raw material, the temperature is reduced to room temperature, water (10mL) is added for quenching, ethyl acetate (4mL) is extracted for 3 times, the ethyl acetate phase is washed by saturated ammonium chloride solution (4mL) and saturated sodium bicarbonate solution (4mL), dried by anhydrous sodium sulfate, filtered by suction, and subjected to vacuum concentration and column chromatography (ethyl acetate (V) and petroleum ether (V) are 4:1) to obtain 3- (4-phenoxyphenyl) -1H- [3- 14 C]Pyrazolo [3,4-d]Pyrimidin-4-amine (6, 7.4. mu. Ci, radiochemical yield 62%). 1 H NMR(400MHz,DMSO-d 6 )δ:13.54(s,1H),8.22(s,1H),7.67(d,J＝8.6Hz,2H),7.43(t,J＝7.9Hz,2H),7.22–7.08(m,5H).ESI-MS m/z:304[M+H] + ,306[M+2+H] + 。
S6: under the protection of argon and at-5 ℃, 3- (4-phenoxyphenyl) -1H- [3- 14 C]Pyrazolo [3,4-d]Pyrimidine-4-amine (6, 10.0. mu. Ci), (S) -1-tert-Butoxycarbonyl-3-hydroxypiperidine (0.55mmol), triphenylphosphine (0.55mmol) and tetrahydrofuran (4mL) were dissolved in water, and diisopropyl azodicarboxylate (0.55mmol) in tetrahydrofuran (1mL) was added dropwise under ice-bath, followed by stirring at-5 ℃ for 3 hours; after the radioactive raw materials completely react, adding water (5mL) for quenching, adding saturated sodium bicarbonate to adjust the pH value to 8-9, extracting with dichloromethane (5mL) for 3 times, drying with anhydrous sodium sulfate, performing suction filtration, performing reduced pressure concentration, and purifying by column chromatography (ethyl acetate) to obtain (R) -3- (4-amino-3- (4-phenoxyphenyl) -1H- [3- 14 C]Pyrazolo [3,4-d]Pyrimidin-1-yl) piperidine-1-carboxylic acid tert-butyl ester (7, 5.7. mu. Ci, radiochemical yield 57%). 1 H NMR(400MHz,DMSO-d 6 )δ:8.25(s,1H),7.66(d,J＝8.4Hz,2H),7.44(t,J＝8.0Hz,2H),7.18–7.12(m,3H),7.08(d,J＝8.0Hz,2H),4.70–4.6(m,1H),4.01–3.5(m,3H),4.12–4.06(m,1H),2.26–2.16(m,1H),2.09–2.03(m,1H),1.61–1.50(m,1H),1.49–1.22(m,9H).ESI-MS m/z:487[M+H] + ,489[M+2+H] + 。
S7: under the protection of argon and at 0 ℃, reacting (R) -3- (4-amino-3- (4-phenoxyphenyl) -1H- [3- 14 C]Pyrazolo [3,4-d]Dissolving tert-butyl pyrimidin-1-yl) piperidine-1-carboxylate (7,6.0 mu Ci) in tetrahydrofuran (4mL), dropwise adding concentrated hydrochloric acid (0.5mL), placing in an oil bath, heating to 40 ℃, and stirring for 1 h; after the radioactive raw material is completely reacted, adding saturated sodium bicarbonate to adjust the pH value to 8-9, extracting with dichloromethane (5mL) for 3 times, drying with anhydrous sodium sulfate, filtering, and concentrating under reduced pressure to obtain (R) -3- (4-phenoxyphenyl) -1- (piperidine-3-yl) -1H- [3- 14 C]Pyrazolo [3,4-d]Pyrimidin-4-amine (8, 5.7. mu. Ci,38mg, radiochemical yield 95%). 1 H NMR(400MHz,DMSO-d 6 )δ:8.23(s,1H),7.65(d,J＝8.6Hz,2H),7.42(t,J＝8.0Hz,2H),7.21–7.08(m,5H),4.75–4.58(m,1H),3.07(d,J＝9.1Hz,1H),2.92(dd,J＝21.4,10.2Hz,2H),2.46(d,J＝12.0Hz,1H),2.16–2.00(m,2H),1.74(d,J＝12.8Hz,1H),1.55(dd,J＝25.0,12.4Hz,1H).ESI-MS m/z:387[M+H] + ,389[M+2+H] + 。
S8: under the protection of argon at 0 deg.C, (R) -3- (4-phenoxyphenyl) -1- (piperidin-3-yl) -1H- [3- 14 C]Pyrazolo [3,4-d]Pyrimidin-4-amine (8, 6.0. mu. Ci) and triethylamine (0.26mmol) were dissolved in methylene chloride (4mL), and allylchloride (0.13mmol) was added dropwise while cooling on ice, and the mixture was stirred at 0 ℃3 h; after the radioactive raw material completely reacts, water (4mL) is added for quenching, dichloromethane (5mL) is used for extraction for 3 times, saturated ammonium chloride is used for washing (3mL), anhydrous sodium sulfate is used for drying, suction filtration and reduced pressure concentration are carried out, and (R) -1- (3- (4-amino-3- (4-phenoxyphenyl) -1H- [3- 14 C]Pyrazolo [3,4-d]Pyrimidin-1-yl) piperidin-1-yl) prop-2-en-1-one (9, 5.2. mu. Ci, radiochemical yield 87%). The conventional method analysis shows that the chemical purity of the target substance carbon-14 labeled ibrutinib (9) is more than 98 percent, and the radiochemical purity is more than 99 percent. 1 H NMR(400MHz,DMSO-d 6 )δ:8.26(s,2H),7.66(d,J＝7.3Hz,2H),7.43(t,J＝7.9Hz,2H),7.22–7.09(m,5H),6.89–6.83(m,0.5H),6.76–6.66(m,0.5H),6.10(dd,J＝26.4,16.7Hz,2H),5.71(d,J＝10.8Hz,0.5H),5.59(d,J＝10.2Hz,0.5H),4.79–4.63(m,1H),4.59–4.50(m,0.5H),4.26–4.14(s,1H),4.05(d,J＝10.9Hz,0.5H),3.74–3.66(m,0.5H),3.25-3.14(m,1H),3.05–2.95(s,0.5H),2.33-2.19(m,1H),2.18–2.06(m,1H),1.92–1.89(m,1H),1.66–1.51(m,1H).ESI-MS m/z:441[M+H] + ,443[M+2+H] + 。
Example 3:
s1: under the protection of argon gas and at room temperature, concentrated sulfuric acid (5mL,18.4M) was slowly dropped into the radioactive raw material [2 ] 14 C]In barium carbonate (28.8. mu. Ci,0.5mmol), escape [2 ] 14 C]CO 2 A tetrahydrofuran solution (5mL) of the metal reagent 4-phenoxyphenylmagnesium bromide (0.55mmol) was introduced through a thin glass tube. Stirring for 30min, reacting, slowly heating to room temperature, adding water (5mL) under cooling in ice-water bath to quench reaction, extracting water phase with dichloromethane, adjusting pH of residual water phase to 4 with dilute hydrochloric acid (1M), extracting with dichloromethane (4mL) for 5 times, mixing organic phases, drying with anhydrous sodium sulfate, vacuum filtering, concentrating under reduced pressure, and drying to obtain 4-phenoxybenzene [2 ] 14 C]Formic acid (2, 27.4. mu. Ci, radiochemical yield 95%). 1 H NMR(400MHz,DMSO-d 6 )δ:12.82(s,1H),7.94(d,J＝8.7Hz,2H),7.46(t,J＝7.8Hz,2H),7.24(t,J＝7.4Hz,1H),7.15–7.09(m,2H),7.02(d,J＝8.7Hz,2H).ESI-MS m/z:214[M-H] - ,215[M+2-H] - 。
S2: under the protection of argon and at 0 ℃, 4-phenoxybenzene [2 ] 14 C]Formic acid (2, 20.0. mu. Ci) was added to a dry Schlenk reaction tube and thionyl chloride was added(2mL), placing the reaction tube in an oil bath, heating to 80 ℃, stirring for 1h, and removing excess thionyl chloride by reduced pressure evaporation; the preparation of the acid chloride and sodium hydride (0.69mmol) were dissolved in tetrahydrofuran (4mL) under argon at 30 ℃ and a solution of malononitrile (0.43mmol) in tetrahydrofuran (2mL) was added dropwise and stirred for 3 h. Radio-TLC analysis shows that after radioactive raw material is completely consumed, water (5mL) is added for quenching, diluted hydrochloric acid is added for adjusting to be neutral, ethyl acetate (4mL) is extracted for 5 times, the ethyl acetate phase is dried by anhydrous sodium sulfate, is filtered by suction, and is subjected to reduced pressure concentration column chromatography (ethyl acetate (V): petroleum ether (V): 1:4) to obtain 2- (hydroxy- (4-phenoxyphenyl) [2- 14 C]Methylene) malononitrile (3, 15.2. mu. Ci, radiochemical yield 76%). 1 H NMR(400MHz,DMSO-d 6 )δ:7.66–7.60(m,2H),7.46–7.39(m,2H),7.18(t,J＝7.4Hz,1H),7.10–7.04(m,2H),6.95(dd,J＝8.5,6.6Hz,2H).ESI-MS m/z:261[M-H] - ,263[M+2-H] - 。
S3: 2- (hydroxy- (4-phenoxyphenyl) [2- 14 C]Methylene) malononitrile (3, 15.0. mu. Ci), diisopropylethylamine (0.40mmol) were dissolved in acetonitrile (2mL), 1M trimethylsilyl diazomethane in tetrahydrofuran (0.40mL) was added dropwise, and stirring was carried out at 30 ℃ for 12 hours; Radio-TLC analysis shows that after radioactive raw material is completely consumed, water (5mL) is added for quenching, diluted hydrochloric acid is added for adjusting the pH value to 6, ethyl acetate (5mL) is added for extraction for 3 times, liquid separation is carried out, the ethyl acetate phase is dried by anhydrous sodium sulfate, suction filtration is carried out, and reduced pressure concentration column chromatography is carried out (ethyl acetate (V): petroleum ether (V): 1:4) to obtain 2- (methoxy- (4-phenoxyphenyl) [ 2-) 14 C]Methylene) malononitrile (4, 12.4. mu. Ci, radiochemical yield 83%). 1 H NMR(400MHz,DMSO-d 6 )δ:7.71(d,J＝8.6Hz,2H),7.49(t,J＝7.9Hz,2H),7.28(t,J＝7.4Hz,1H),7.16(t,J＝9.0Hz,4H),3.93(s,3H).ESI-MS m/z:277[M+H] + ,279[M+2+H] + 。
S4: under the protection of argon, 2- (methoxy- (4-phenoxyphenyl) [2- 14 C]Methylene) malononitrile (4,12.0 mu Ci) and hydrazine hydrate (0.446mmol) are dissolved in propanol (4mL), placed in an oil bath, heated to 100 ℃ and stirred for 1.5 h; TLC monitoring radioactive starting material reaction completion, adding water (1mL) to quench, removing solvent under reduced pressure, extracting with ethyl acetate (5mL) for 3 times, and adding saturated ammonium chloride solution (2 m) to the ethyl acetate phaseL) and a saturated sodium bicarbonate solution (2mL), followed by drying over anhydrous sodium sulfate, suction filtration, and concentration under reduced pressure for column chromatography (ethyl acetate (V): petroleum ether (V) ═ 1:1) to obtain 5-amino-3- (4-phenoxyphenyl) -1H- [3- 14 C]Pyrazole-4-carbonitrile (5, 8.5. mu. Ci, radiochemical yield 71%). 1 H NMR(400MHz,DMSO-d 6 )δ:12.11(s,1H),7.80(d,J＝8.6Hz,2H),7.43(dd,J＝17.3,9.4Hz,2H),7.17(t,J＝7.4Hz,1H),7.14-7.06(m,4H),6.46(s,2H).ESI-MS m/z:277[M+H] + ,279[M+2+H] + 。
S5: under the protection of argon, 5-amino-3- (4-phenoxyphenyl) -1H- [3- 14 C]Dissolving pyrazole-4-cyanogen (5,12.0 mu Ci) in formamide (4mL), placing in an oil bath, heating to 170 ℃, and stirring for 2 h; TLC (thin layer chromatography) is used for monitoring the completion of the reaction of the radioactive raw material, the temperature is reduced to room temperature, water (10mL) is added for quenching, ethyl acetate (4mL) is extracted for 3 times, the ethyl acetate phase is washed by saturated ammonium chloride solution (4mL) and saturated sodium bicarbonate solution (4mL) in sequence, dried by anhydrous sodium sulfate, filtered, decompressed and concentrated for column chromatography (ethyl acetate (V): petroleum ether (V): 4:1) to obtain 3- (4-phenoxyphenyl) -1H- [3- 14 C]Pyrazolo [3,4-d]Pyrimidin-4-amine (6, 9.7. mu. Ci, radiochemical yield 81%). 1 H NMR(400MHz,DMSO-d 6 )δ:13.54(s,1H),8.22(s,1H),7.67(d,J＝8.6Hz,2H),7.43(t,J＝7.9Hz,2H),7.22–7.08(m,5H).ESI-MS m/z:304[M+H] + ,306[M+2+H] + 。
S6: under the protection of argon and cooling in ice bath, 3- (4-phenoxyphenyl) -1H- [3- 14 C]Pyrazolo [3,4-d]Pyrimidin-4-amine (6, 10.0. mu. Ci), (S) -1-tert-butoxycarbonyl-3-hydroxypiperidine (0.55mmol), triphenylphosphine (0.55mmol) and tetrahydrofuran (4mL) dissolved in diisopropyl azodicarboxylate (0.55mmol) in tetrahydrofuran (1mL) was added dropwise and stirred at room temperature for 1 h; after the radioactive raw materials completely react, adding water (5mL) for quenching, adding saturated sodium bicarbonate to adjust the pH value to 8-9, extracting with dichloromethane (5mL) for 3 times, drying with anhydrous sodium sulfate, performing suction filtration, performing reduced pressure concentration, and purifying by column chromatography (ethyl acetate) to obtain (R) -3- (4-amino-3- (4-phenoxyphenyl) -1H- [3- 14 C]Pyrazolo [3,4-d]Pyrimidin-1-yl) piperidine-1-carboxylic acid tert-butyl ester (7, 6.9. mu. Ci, radiochemical yield 69%). 1 H NMR(400MHz,DMSO-d 6 )δ:8.25(s,1H),7.66(d,J＝8.4Hz,2H),7.44(t,J＝8.0Hz,2H),7.18–7.12(m,3H),7.08(d,J＝8.0Hz,2H),4.70–4.6(m,1H),4.01–3.5(m,3H),4.12–4.06(m,1H),2.26–2.16(m,1H),2.09–2.03(m,1H),1.61–1.50(m,1H),1.49–1.22(m,9H).ESI-MS m/z:487[M+H] + ,489[M+2+H] + 。
S7: under the protection of argon and cooling in ice bath, (R) -3- (4-amino-3- (4-phenoxyphenyl) -1H- [ 3-) 14 C]Pyrazolo [3,4-d]Pyrimidin-1-yl) piperidine-1-carboxylic acid tert-butyl ester (7, 6.0. mu. Ci) was dissolved in tetrahydrofuran (4mL), concentrated hydrochloric acid (0.5mL) was added dropwise, and the mixture was stirred at 30 ℃ for 3 hours; after the radioactive raw material is completely reacted, adding saturated sodium bicarbonate to adjust the pH value to 8-9, extracting with dichloromethane (5mL) for 3 times, drying with anhydrous sodium sulfate, filtering, and concentrating under reduced pressure to obtain (R) -3- (4-phenoxyphenyl) -1- (piperidine-3-yl) -1H- [3- 14 C]Pyrazolo [3,4-d]Pyrimidin-4-amine (8, 5.4. mu. Ci, radiochemical yield 90%). 1 H NMR(400MHz,DMSO-d 6 )δ:8.23(s,1H),7.65(d,J＝8.6Hz,2H),7.42(t,J＝8.0Hz,2H),7.21–7.08(m,5H),4.75–4.58(m,1H),3.07(d,J＝9.1Hz,1H),2.92(dd,J＝21.4,10.2Hz,2H),2.46(d,J＝12.0Hz,1H),2.16–2.00(m,2H),1.74(d,J＝12.8Hz,1H),1.55(dd,J＝25.0,12.4Hz,1H).ESI-MS m/z:387[M+H] + ,389[M+2+H] + 。
S8: under the protection of argon and under the cooling of ice bath, (R) -3- (4-phenoxyphenyl) -1- (piperidin-3-yl) -1H- [3- 14 C]Pyrazolo [3,4-d]Dissolving pyrimidin-4-amine (8, 6.0. mu. Ci) and triethylamine (0.26mmol) in dichloromethane (4mL), dropwise adding allyl chloride (0.13mmol) in ice bath, and stirring at 30 ℃ for 40 min; after the radioactive raw material completely reacts, water (4mL) is added for quenching, dichloromethane (5mL) is used for extraction for 3 times, saturated ammonium chloride is used for washing (3mL), anhydrous sodium sulfate is used for drying, suction filtration and reduced pressure concentration are carried out, and (R) -1- (3- (4-amino-3- (4-phenoxyphenyl) -1H- [3- 14 C]Pyrazolo [3,4-d]Pyrimidin-1-yl) piperidin-1-yl) prop-2-en-1-one (9, 5.3. mu. Ci, radiochemical yield 88%). The conventional method analysis shows that the chemical purity of the target substance carbon-14 labeled ibrutinib (9) is more than 98 percent, and the radiochemical purity is more than 99 percent. 1 H NMR(400MHz,DMSO-d 6 )δ:8.26(s,2H),7.66(d,J＝7.3Hz,2H),7.43(t,J＝7.9Hz,2H),7.22–7.09(m,5H),6.89–6.83(m,0.5H),6.76–6.66(m,0.5H),6.10(dd,J＝26.4,16.7Hz,2H),5.71(d,J＝10.8Hz,0.5H),5.59(d,J＝10.2Hz,0.5H),4.79–4.63(m,1H),4.59–4.50(m,0.5H),4.26–4.14(s,1H),4.05(d,J＝10.9Hz,0.5H),3.74–3.66(m,0.5H),3.25-3.14(m,1H),3.05–2.95(s,0.5H),2.33-2.19(m,1H),2.18–2.06(m,1H),1.92–1.89(m,1H),1.66–1.51(m,1H).ESI-MS m/z:441[M+H] + ,443[M+2+H] + .
Example 4:
s1: under the protection of argon gas and at room temperature, concentrated sulfuric acid (5mL,18.4M) was slowly dropped into the radioactive raw material [2 ] 14 C]In barium carbonate (28.8. mu. Ci,0.5mmol), escape [2 ] 14 C]CO 2 A tetrahydrofuran solution (5mL) of the metal reagent 4-phenoxyphenylmagnesium chloride (0.55mmol) was introduced through a thin glass tube. Stirring for 30min, reacting, slowly heating to room temperature, adding water (5mL) under cooling in ice-water bath to quench reaction, extracting water phase with dichloromethane, adjusting pH of residual water phase to 4 with dilute hydrochloric acid (1M), extracting with dichloromethane (4mL) for 5 times, mixing organic phases, drying with anhydrous sodium sulfate, vacuum filtering, concentrating under reduced pressure, and drying to obtain 4-phenoxybenzene [2 ] 14 C]Formic acid (2, 25.6. mu. Ci, radiochemical yield 89%). 1 H NMR(400MHz,DMSO-d 6 )δ:12.82(s,1H),7.94(d,J＝8.7Hz,2H),7.46(t,J＝7.8Hz,2H),7.24(t,J＝7.4Hz,1H),7.15–7.09(m,2H),7.02(d,J＝8.7Hz,2H).ESI-MS m/z:214[M-H] - ,215[M+2-H] - 。
S2: under the protection of argon and at 0 ℃, 4-phenoxybenzene [2 ] 14 C]Formic acid (2, 20.0. mu. Ci) was added to a dry Schlenk reaction tube, and oxalyl chloride in dichloromethane (0.5mL,0.1M) and DMF in dichloromethane (0.2mL,0.01M) were added; stirring at room temperature for 1h, and removing the solvent by evaporation under reduced pressure; the preparation of the acid chloride, sodium hydride (0.69mmol) was dissolved in tetrahydrofuran (4mL) under argon atmosphere at 0 deg.C, and a solution of malononitrile (0.43mmol) in tetrahydrofuran (2mL) was added dropwise and stirred at room temperature overnight. Radio-TLC analysis shows that after radioactive raw material is completely consumed, water (5mL) is added for quenching, diluted hydrochloric acid is added for adjusting to be neutral, ethyl acetate (4mL) is extracted for 5 times, the ethyl acetate phase is dried by anhydrous sodium sulfate, is filtered by suction, and is subjected to reduced pressure concentration column chromatography (ethyl acetate (V): petroleum ether (V): 1:4) to obtain 2- (hydroxy- (4-phenoxyphenyl) [2- 14 C]Methylene) malononitrile (3, 15.6. mu. Ci, radiochemical yield 78%). 1 H NMR(400MHz,DMSO-d 6 )δ:7.66–7.60(m,2H),7.46–7.39(m,2H),7.18(t,J＝7.4Hz,1H),7.10–7.04(m,2H),6.95(dd,J＝8.5,6.6Hz,2H).ESI-MS m/z:261[M-H] - ,263[M+2-H] - 。
S3: 2- (hydroxy- (4-phenoxyphenyl) [2- 14 C]Methylene) malononitrile (3, 15.0. mu. Ci) and triethylamine (0.40mmol) were dissolved in acetonitrile (2mL), and 1M trimethylsilyl diazomethane in tetrahydrofuran (0.40mL) was added dropwise and stirred at room temperature for 12 hours; Radio-TLC analysis shows that after radioactive raw material is completely consumed, water (5mL) is added for quenching, diluted hydrochloric acid is added for adjusting the pH value to 6, ethyl acetate (5mL) is added for extraction for 3 times, liquid separation is carried out, the ethyl acetate phase is dried by anhydrous sodium sulfate, suction filtration is carried out, and reduced pressure concentration column chromatography is carried out (ethyl acetate (V): petroleum ether (V): 1:4) to obtain 2- (methoxy- (4-phenoxyphenyl) [ 2-) 14 C]Methylene) malononitrile (4, 11.2. mu. Ci, radiochemical yield 75%). 1 H NMR(400MHz,DMSO-d 6 )δ:7.71(d,J＝8.6Hz,2H),7.49(t,J＝7.9Hz,2H),7.28(t,J＝7.4Hz,1H),7.16(t,J＝9.0Hz,4H),3.93(s,3H).ESI-MS m/z:277[M+H] + ,279[M+2+H] + 。
S4: under the protection of argon, 2- (methoxy- (4-phenoxyphenyl) [2- 14 C]Methylene) malononitrile (4,12.0 mu Ci) and hydrazine hydrate (0.446mmol) are dissolved in ethanol (4mL), and the solution is placed in an oil bath to be heated to 100 ℃ and stirred for 2 h; TLC (thin layer chromatography) is used for monitoring the completion of the reaction of the radioactive raw material, water (1mL) is added for quenching, the solvent is removed by rotation under reduced pressure, ethyl acetate (5mL) is extracted for 3 times, the ethyl acetate phase is washed by saturated ammonium chloride solution (2mL) and saturated sodium bicarbonate solution (2mL) in turn, dried by anhydrous sodium sulfate, filtered by suction, and subjected to column chromatography under reduced pressure (ethyl acetate (V): petroleum ether (V): 1) to obtain 5-amino-3- (4-phenoxyphenyl) -1H- [3- 14 C]Pyrazole-4-carbonitrile (5, 7.4. mu. Ci, radiochemical yield 62%). 1 H NMR(400MHz,DMSO-d 6 )δ:12.11(s,1H),7.80(d,J＝8.6Hz,2H),7.43(dd,J＝17.3,9.4Hz,2H),7.17(t,J＝7.4Hz,1H),7.14-7.06(m,4H),6.46(s,2H).ESI-MS m/z:277[M+H] + ,279[M+2+H] + 。
S5: under the protection of argon, 5-amino-3- (4-phenoxyphenyl) -1H- [3- 14 C]Pyrazole-4-carbonitrile (5, 12.0. mu. Ci) was dissolved in formamide (4mL) and placed in an oil bath and warmed to room temperatureStirring for 0.5h at 200 ℃; TLC (thin layer chromatography) is used for monitoring the completion of the reaction of the radioactive raw material, the temperature is reduced to room temperature, water (10mL) is added for quenching, ethyl acetate (4mL) is extracted for 3 times, the ethyl acetate phase is washed by saturated ammonium chloride solution (4mL) and saturated sodium bicarbonate solution (4mL) in sequence, dried by anhydrous sodium sulfate, filtered, decompressed and concentrated for column chromatography (ethyl acetate (V): petroleum ether (V): 4:1) to obtain 3- (4-phenoxyphenyl) -1H- [3- 14 C]Pyrazolo [3,4-d]Pyrimidin-4-amine (6, 9.2. mu. Ci, radiochemical yield 77%). 1 H NMR(400MHz,DMSO-d 6 )δ:13.54(s,1H),8.22(s,1H),7.67(d,J＝8.6Hz,2H),7.43(t,J＝7.9Hz,2H),7.22–7.08(m,5H).ESI-MS m/z:304[M+H] + ,306[M+2+H] + 。
S6: under the protection of argon, 3- (4-phenoxyphenyl) -1H- [3- 14 C]Pyrazolo [3,4-d]Pyrimidin-4-amine (6, 10.0. mu. Ci), (S) -1-tert-butoxycarbonyl-3-hydroxypiperidine (0.55mmol), triphenylphosphine (0.55mmol) and tetrahydrofuran (4mL) dissolved in tetrahydrofuran, diisopropyl azodicarboxylate (0.55mmol) was added dropwise in tetrahydrofuran (1mL) under ice-bath, and the mixture was stirred at room temperature for 1 hour; after the radioactive raw materials completely react, adding water (5mL) for quenching, adding saturated sodium bicarbonate to adjust the pH value to 8-9, extracting with dichloromethane (5mL) for 3 times, drying with anhydrous sodium sulfate, performing suction filtration, performing reduced pressure concentration, and purifying by column chromatography (ethyl acetate) to obtain (R) -3- (4-amino-3- (4-phenoxyphenyl) -1H- [3- 14 C]Pyrazolo [3,4-d]Pyrimidin-1-yl) piperidine-1-carboxylic acid tert-butyl ester (7, 6.5. mu. Ci,55mg, radiochemical yield 65%). 1 H NMR(400MHz,DMSO-d 6 )δ:8.25(s,1H),7.66(d,J＝8.4Hz,2H),7.44(t,J＝8.0Hz,2H),7.18–7.12(m,3H),7.08(d,J＝8.0Hz,2H),4.70–4.6(m,1H),4.01–3.5(m,3H),4.12–4.06(m,1H),2.26–2.16(m,1H),2.09–2.03(m,1H),1.61–1.50(m,1H),1.49–1.22(m,9H).ESI-MS m/z:487[M+H] + ,489[M+2+H] + 。
S7: under the protection of argon, (R) -3- (4-amino-3- (4-phenoxyphenyl) -1H- [3- 14 C]Pyrazolo [3,4-d]Dissolving tert-butyl pyrimidin-1-yl) piperidine-1-carboxylate (7, 6.0. mu. Ci) in dichloromethane (4mL), dropwise adding trifluoroacetic acid (0.1mL) under cooling in an ice bath, placing in an oil bath, heating to 40 ℃, and stirring for 1 h; after the radioactive raw materials completely react, adding saturated sodium bicarbonate to adjust the pH value to 8-9, extracting with dichloromethane (5mL) for 3 times, and drying with anhydrous sodium sulfateDrying, suction filtering, vacuum concentrating to obtain (R) -3- (4-phenoxyphenyl) -1- (piperidine-3-yl) -1H- [3- 14 C]Pyrazolo [3,4-d]Pyrimidin-4-amine (8, 5.4. mu. Ci, radiochemical yield 90%). 1 H NMR(400MHz,DMSO-d 6 )δ:8.23(s,1H),7.65(d,J＝8.6Hz,2H),7.42(t,J＝8.0Hz,2H),7.21–7.08(m,5H),4.75–4.58(m,1H),3.07(d,J＝9.1Hz,1H),2.92(dd,J＝21.4,10.2Hz,2H),2.46(d,J＝12.0Hz,1H),2.16–2.00(m,2H),1.74(d,J＝12.8Hz,1H),1.55(dd,J＝25.0,12.4Hz,1H).ESI-MS m/z:387[M+H] + ,389[M+2+H] + 。
S8: under the protection of argon and cooling in ice bath, (R) -3- (4-phenoxyphenyl) -1- (piperidin-3-yl) -1H- [3- 14 C]Pyrazolo [3,4-d]Dissolving pyrimidine-4-amine (8,6.0 mu Ci) and triethylamine (0.26mmol) in dichloromethane (4mL), and dropwise adding allyl chloride (0.13 mmol); heating to 30 ℃ and stirring for 1 h; after the radioactive raw material completely reacts, water (4mL) is added for quenching, dichloromethane (5mL) is used for extraction for 3 times, saturated ammonium chloride is used for washing (3mL), anhydrous sodium sulfate is used for drying, suction filtration and reduced pressure concentration are carried out, and (R) -1- (3- (4-amino-3- (4-phenoxyphenyl) -1H- [3- 14 C]Pyrazolo [3,4-d]Pyrimidin-1-yl) piperidin-1-yl) prop-2-en-1-one (9, 5.5. mu. Ci, radiochemical yield 92%). The conventional method analysis shows that the chemical purity of the target substance carbon-14 labeled ibrutinib (9) is more than 98 percent, and the radiochemical purity is more than 99 percent. 1 H NMR(400MHz,DMSO-d 6 )δ:8.26(s,2H),7.66(d,J＝7.3Hz,2H),7.43(t,J＝7.9Hz,2H),7.22–7.09(m,5H),6.89–6.83(m,0.5H),6.76–6.66(m,0.5H),6.10(dd,J＝26.4,16.7Hz,2H),5.71(d,J＝10.8Hz,0.5H),5.59(d,J＝10.2Hz,0.5H),4.79–4.63(m,1H),4.59–4.50(m,0.5H),4.26–4.14(s,1H),4.05(d,J＝10.9Hz,0.5H),3.74–3.66(m,0.5H),3.25-3.14(m,1H),3.05–2.95(s,0.5H),2.33-2.19(m,1H),2.18–2.06(m,1H),1.92–1.89(m,1H),1.66–1.51(m,1H).ESI-MS m/z:441[M+H] + ,443[M+2+H] + 。
Many modifications may be made by one of ordinary skill in the art in light of the above teachings. Therefore, it is intended that the invention not be limited to the particular details of the embodiments disclosed, but that the invention will include all embodiments falling within the scope of the appended claims.

Claims (6)

1. A method for synthesizing radioisotope carbon-14 labeled ibrutinib, comprising the steps of:
s1: under the protection of inert gas and at the low temperature of < -70 ℃, 4-phenoxyphenyl lithium (1) is reacted with sodium benzoate 14 C]Barium carbonate released [ alpha ], [ alpha ] and [ alpha ], [ alpha ] barium carbonate 14 C]CO 2 Reaction preparation of 4-phenoxybenzene [ alpha ], [ beta ] -pheno ] or a mixture thereof 14 C]Formic acid (2);
s2: under the protection of inert gas and at the reaction temperature of 0-80 ℃, 4-phenoxybenzene [ alpha ], [ alpha ] an 14 C]The formic acid (2) reacts with thionyl chloride or oxalyl chloride to produce 4-phenoxybenzene [2 ] 14 C]Formyl chloride; reacting the obtained acyl chloride with sodium hydrogen and malononitrile in tetrahydrofuran for 3-15 h, and performing conventional post-treatment and column chromatography purification to obtain the 2- (hydroxy- (4-phenoxyphenyl), [2 ] 14 C]Methylene) malononitrile (3);
s3: under the protection of inert gas and at the reaction temperature of 0-30 ℃, the intermediate 2- (hydroxy- (4-phenoxyphenyl) [ ] 14 C]Methylene) malononitrile (3), trimethylsilyldiazomethane and organic alkali are stirred in acetonitrile for 12-48 h, and the reaction is finished; adjusting the reaction solution to weak acidity (pH about 6) with dilute hydrochloric acid, and performing conventional post-treatment and column chromatography purification to obtain 2- (methoxy- (4-phenoxyphenyl) ([ 2 ]) 14 C]Methylene) malononitrile (4);
s4: under the protection of inert gas and at the reaction temperature of 50-100 ℃, the intermediate 2- (methoxy- (4-phenoxyphenyl), [2 ] 14 C]Methylene) malononitrile (4) and hydrazine hydrate are stirred in lower alcohol for 1-3 h; obtaining 5-amino-3- (4-phenoxyphenyl) -1 through conventional post-treatment and column chromatography purificationH-[3- 14 C]Pyrazole-4-cyanogen (5), wherein the lower alcohol is selected from one or more of methanol, ethanol or propanol;
s5: under the protection of inert gas and at the reaction temperature of 80-200 ℃, the intermediate 5-amino-3- (4-phenoxyphenyl) -1H-[3- 14 C]Pyrazole-4-cyanide (5) is reacted with formamide; after the reaction is finished, the reaction mixture is subjected to conventional post-treatment to obtain an intermediate 3- (4-phenoxyphenyl) -1H-[3- 14 C]Pyrazolo [3,4-d]Pyrimidin-4-amine (6);
s6: under the protection of inert gas and the low-temperature reaction at-5 to 30 ℃, the intermediate 3- (4-phenoxyphenyl) -1H-[3- 14 C]Pyrazolo [3,4-d]Pyrimidin-4-amine (6) with (A)S) -reacting 1-tert-butoxycarbonyl-3-hydroxypiperidine, triphenylphosphine and azodicarboxylic acid diisocynate in tetrahydrofuran; after the reaction is finished, the product is obtained by conventional post-treatment and column chromatography purificationR) -3- (4-amino-3- (4-phenoxyphenyl) -1H-[3- 14 C]Pyrazolo [3,4-d]Pyrimidin-1-yl) piperidine-1-carboxylic acid tert-butyl ester (7);
s7: at a reaction temperature of 10 to 40 ℃, (b) an intermediateR) -3- (4-amino-3- (4-phenoxyphenyl) -1H-[3- 14 C]Pyrazolo [3,4-d]Removing a protecting group from tert-butyl pyrimidin-1-yl) piperidine-1-carboxylate (7) in an acidic tetrahydrofuran solution or a dichloromethane solution; after the reaction is finished, the product is obtained by conventional post-treatment and column chromatography purificationR) -3- (4-phenoxyphenyl) -1- (piperidin-3-yl) -1H-[3- 14 C]Pyrazolo [3,4-d]Pyrimidin-4-amine (8);
s8: under the protection of inert gas and at the reaction temperature of 0-30 ℃, an intermediate (A)R) -3- (4-phenoxyphenyl) -1- (piperidin-3-yl) -1H-[3- 14 C]Pyrazolo [3,4-d]Reacting pyrimidine-4-amine (8), triethylamine and allyl acyl chloride in dichloromethane; after the reaction is finished, the product is obtained by conventional post-treatment and column chromatography purificationR) -1- (3- (4-amino-3- (4-phenoxyphenyl) -1H-[3- 14 C]Pyrazolo [3,4-d]Pyrimidin-1-yl) piperidin-1-yl) prop-2-en-1-one (9).
2. The method of synthesis according to claim 1, characterized in that: the inert gas is argon or nitrogen.
3. The method of synthesis according to claim 1, characterized in that: in S1, 4-phenoxybenzene [ alpha ], [ beta ], [ alpha ], [ beta ], [ alpha ], [ beta ], [ 14 C]The formic acid (2) can also be reacted with the Grignard reagent 4-phenoxyphenylmagnesium halide under protection of an inert gas at room temperature 14 C]Preparation of (2) by reaction of CO2, wherein the reaction is performed in the presence of a catalyst 14 C]When CO2 is completely consumed, the reaction is finished, and 4-phenoxybenzene can be obtained by conventional post treatment[ 14 C]Formic acid (2).
4. The method of synthesis according to claim 3, characterized in that: in S2, the halogen atom of the Grignard reagent 4-phenoxyphenylmagnesium halide is selected from one of Cl, Br and I.
5. The method of synthesis according to claim 1, characterized in that: in S3, the organic base is selected from one or both of diisopropylethylamine or triethylamine.
6. The synthetically synthesized radioisotopic carbon-14-labeled ibrutinib of claims 1-5, wherein the radioisotopic carbon-14 is labeled at the 3-position of a pyrazolo [3,4-d ] pyrimidine fragment of the ibrutinib molecule and has the following structural formula:
Figure DEST_PATH_IMAGE001
(. indicates the carbon-14 marker site).
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Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20140228340A1 (en) * 2011-09-09 2014-08-14 Merck Patent Gmbh Benzonitrile derivatives as kinase inhibitors
US20160346200A1 (en) * 2014-02-07 2016-12-01 Auspex Pharmaceuticals, Inc. Novel pharmaceutical formulations
US20170035881A1 (en) * 2015-10-19 2017-02-09 Acerta Pharma B.V. Therapeutic Combinations of an IRAK4 Inhibitor and a BTK Inhibitor
US20180251489A1 (en) * 2015-08-28 2018-09-06 Cornell University Cornell Center for Technology Enterprise & Commercialization (CCTEC) Malt1 inhibitors and uses thereof
CN109956926A (en) * 2017-12-26 2019-07-02 上海启甄环境科技有限公司 A kind of radioactive isotope carbon -14 marks the synthetic method of imidacloprid
CN111116593A (en) * 2019-11-27 2020-05-08 河北合佳医药科技集团股份有限公司 Continuous and low-cost preparation method of imatinib

Patent Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20140228340A1 (en) * 2011-09-09 2014-08-14 Merck Patent Gmbh Benzonitrile derivatives as kinase inhibitors
US20160346200A1 (en) * 2014-02-07 2016-12-01 Auspex Pharmaceuticals, Inc. Novel pharmaceutical formulations
US20180251489A1 (en) * 2015-08-28 2018-09-06 Cornell University Cornell Center for Technology Enterprise & Commercialization (CCTEC) Malt1 inhibitors and uses thereof
US20170035881A1 (en) * 2015-10-19 2017-02-09 Acerta Pharma B.V. Therapeutic Combinations of an IRAK4 Inhibitor and a BTK Inhibitor
CN109956926A (en) * 2017-12-26 2019-07-02 上海启甄环境科技有限公司 A kind of radioactive isotope carbon -14 marks the synthetic method of imidacloprid
CN111116593A (en) * 2019-11-27 2020-05-08 河北合佳医药科技集团股份有限公司 Continuous and low-cost preparation method of imatinib

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