CN106467517B

CN106467517B - The Abemaciclib derivative of deuterium modification

Info

Publication number: CN106467517B
Application number: CN201610659847.XA
Authority: CN
Inventors: 张寅生; 柳英帅; 李莉; 苗雷; 徐同杰; 刘海艳; 马雪琴; 俞森; 徐宏江
Original assignee: Chia Tai Tianqing Pharmaceutical Group Co Ltd
Current assignee: Chia Tai Tianqing Pharmaceutical Group Co Ltd
Priority date: 2015-08-14
Filing date: 2016-08-12
Publication date: 2019-09-06
Anticipated expiration: 2036-08-12
Also published as: CN106467517A

Abstract

The invention belongs to field of medicinal chemistry, and in particular to purposes of Abemaciclib derivative, preparation method, the pharmaceutical composition of the Abemaciclib derivative of the modification containing deuterium, the Abemaciclib derivative of deuterium modification and its pharmaceutical composition of deuterium modification in preparation treatment cell proliferation disorders drug.Compared with Abemaciclib, certain compounds of the invention (especially embodiment compound) have superior pharmacokinetic property, are expected to reduce clinical use dosage, to reduce treatment cost to allow more patients to be benefited.

Description

The Abemaciclib derivative of deuterium modification

Technical field

The invention belongs to field of medicinal chemistry, and in particular to the Abemaciclib derivative of a kind of novel deuterium modification, its Preparation method, containing deuterium modification Abemaciclib derivative pharmaceutical composition, deuterium modification Abemaciclib derivative and Purposes of its pharmaceutical composition in the drug of preparation treatment cell proliferation disorders.

Background technique

Cyclin-dependent kinase (cyclin-dependent kinase, CDK) is a kind of serine/threonine kinase Enzyme, the CDK-cyclin compound formed as intracellular important signal transducers and period plain (cyclin) participate in carefully Growth, proliferation, suspend mode or the apoptosis of born of the same parents.

CDK family includes 1-13, and cyclin points are A-L, and different CDK couples different cyclin respectively.Wherein, Cyclin D family starts to express in the G1 phase, in conjunction with and activate CDK4 and CDK6, form CDK4/6-cyclin D compound, make A series of substrate phosphorylations including Retinoblastoma Protein.The activation and the increasing of some tumours of CDK4/6 specificity Grow it is closely related, therefore develop CDK4/6 inhibitor become a kind of targeting therapy on tumor effective means.It has just listed within 2015 New drug Palbociclib is exactly an orally active CDK4/6 inhibitor, the treatment of the current women breast cancer in advanced stage.

Abemaciclib (LY-2835219) is the New Target tropism breast cancer treatment medicine developed by Li Lai drugmaker Object, it can selective depression cell cycle protein dependent kinase 4 and 6 (CDK4/6) and Pim-1 kinases, restore cell week Phase control, blocks tumor cell proliferation, and clinical second phase significant effect is a kind of novel breast cancer and lung cancer therapy drug.

Many drugs limit making in certain diseases for they due to poor absorption, distribution, metabolisming property With.Poor ADME property is also that many drug candidates is caused to fail the reason of passing through clinical test.Although with special system Agent technique or prodrug technologies can improve the ADME property of some drugs in some cases, but deposit to most of drug candidates ADME problem not can effectively solve still, especially for the tachymetabolism problem of drug, cause it is many originally can be efficient The drug of disease is treated to fall from internal metabolite clearance due to too fast and be difficult to patent medicine.

Deuterated modification is to improve a kind of very potential new drug development technology of pharmacokinetic properties.In the method, Try by replacing to slow down the drug metabolism of CYP mediation or reduce undesirable one or more hydrogen atoms progress D-atom Metabolin generation, as a result may significantly extend its drug metabolism circulation, reduce toxic metabolite generation and drug Between interaction, improve safety and obtain more preferably curative effect.However, even if D-atom is mixed with known metabolism position It sets, influence of the deuterium modification for the metabolisming property of drug is still uncertain, it is therefore necessary to by the reality of deuterated molecule Preparation and test, just can determine that deuterated drug and difference of the non-deuterated drug in terms of metabolism.

Summary of the invention

One aspect of the present invention provide a kind of compound or its pharmaceutically acceptable salt shown in formula I, solvate or Prodrug:

Wherein R₁、R₂、R₄、R₅、R₆、R₇、R₈、R₉And R₁₁Separately it is selected from H (hydrogen) or D (deuterium), R₃、R₁₀、R₁₂Respectively Independently selected from CH₃、CH₂D、CHD₂Or CD₃, condition is that compound of formula I contains at least one D-atom and do not include following chemical combination Object:

Preferably, wherein R₁、R₂、R₅、R₆、R₇、R₈Or R₁₁At least one is D.

It is further preferred that wherein R₁、R₂It is simultaneously D.

It is further preferred that wherein R₅、R₆It is simultaneously D.

It is further preferred that wherein R₇、R₈It is simultaneously D.

Still more preferably, wherein R₅、R₆、R₇、R₈It is simultaneously D.

In certain preferred embodiments of the invention, the example of compound shown in formula I is as follows:

In certain particularly preferred embodiments of the invention, the example of compound shown in formula I is as follows:

Term " pharmaceutically acceptable salt " refers to the biological efficacy for remaining the free bronsted lowry acids and bases bronsted lowry of specific compound and does not have There is the salt of biology ill-effect.The example of pharmaceutically acceptable salt includes but is not limited to: (1) acid-addition salts and inorganic acid Such as the salt of the formation such as hydrochloric acid, sulfuric acid, hydrobromic acid, nitric acid, phosphoric acid；Or with organic acids for example malic acid, fumaric acid, maleic acid, Benzoic acid, phenylacetic acid, succinic acid, tartaric acid, citric acid, methanesulfonic acid, ethanesulfonic acid, hydroxyacetic acid, cinnamic acid, pyruvic acid, formic acid, The salt of the formation such as acetic acid, propionic acid, oxalic acid, malonic acid, acrylic acid, mandelic acid；Or (2) base addition salts and alkali metal such as lithium, The salt of the formation such as sodium, potassium；With the salt of the formation such as alkaline-earth metal such as calcium, magnesium；With organic bases for example ammonium, choline, diethanol amine, rely Propylhomoserin, ethylenediamine, tert-butylamine, t-octanylamine, three (methylol) aminomethanes, N- methyl glucose osamine, triethanolamine, dehydrogenated rosin The salt of the formation such as amine.For a person skilled in the art, other pharmaceutically acceptable salts are known.

Term " solvate " refers to form this patent hair of the complex of solid-state or liquid by being coordinated with solvent molecule The form of bright compound.The example of such form is hydrate, alcohol adduct etc..

Term " prodrug " refers to any medicament for being converted to parent drug in vivo.Prodrug is often useful, because, In some cases, they are easier to be administered than parent drug.For example, being administered orally, they are bioavailable, and female Body drug is not but.Prodrug can also improve the solubility in pharmaceutical composition relative to parent drug.Prodrug can be via the side of enzyme Method and the approach of metabolism hydrolysis are converted into parent drug.

If any atom of the compound of institute's labelled synthesis does not specify in the present invention, any of the atom can be represented A kind of stable isotope.Unless stated otherwise, when position a certain in structure is defined as H i.e. hydrogen (H-1), which is contained only Naturally occurring isotopic mass.It equally, unless stated otherwise, should when position a certain in structure is defined as D i.e. deuterium (H-2) Position containing isotopic mass is at least 3340 times bigger than naturally occurring isotopic mass (0.015%) (i.e. at least can the same position of 50.1% deuterium Element).

In the present invention the deuterated rate of the compound of institute's labelled synthesis refer to the isotopic content of labelled synthesis with it is naturally occurring Isotopic mass ratio.Each of compound of institute's labelled synthesis specifies the deuterated rate of D-atom that can be at least in the present invention 3500 times (52.5%), be at least 4000 times (60%), be at least 4500 times (67.5%), be at least 5000 times (75%), extremely It is less 5500 times (82.5%), be at least 6000 times (90%), is at least 6333.3 times (95%), is at least 6466.7 times (97%), it is at least 6566.7 times (98.5%), is at least 6600 times (99%), is at least 6633.3 times (99.5%).

Isotopic body (isotopologues) in the present invention refers to be only had in isotopics not in terms of chemical structure Same compound.The compound chemical structure having the same of institute's labelled synthesis in the present invention is only formed in the atom of its molecule The variation of middle isotope.Therefore, the present invention in institute's labelled synthesis specific position containing deuteride can similarly contain it is considerably less The position hydrogen isotope body, the measurement of hydrogen isotope body of certain position in the present invention in the compound of institute's labelled synthesis determines Many factors, including deuterated reagent (D₂O、D₂、NaBD₄、LiAlD₄Deng) deuterium isotopic purity and introduce deuterium isotope The validity of synthetic method.However, the amount sum of the hydrogen isotope body of certain this position will be less than 49.9% as previously described.This hair The amount sum of the hydrogen isotope body of certain position in the compound of bright middle institute's labelled synthesis will less than 47.5%, 40%, 32.5%, 25%, 17.5%, 10%, 5%, 3%, 1% or 0.5%.

Herein, any each atom for being not designated as deuterium is with the presence of its natural isotopic abundance.

The present invention further provides the illustrative methods for being used to prepare compound and each intermediate shown in formula I.

Route 1 provides a kind of illustrative methods for being used to prepare compound shown in formula I.

As shown in Scheme 1, in the presence of a catalyst, Formula II compound is reacted with double two boron of pinacol is prepared formula III Compound, further coupling reaction obtains formula IV compound under metallic catalyst effect.Finally in the presence of a catalyst, formula IV compound and Formula V compound react to obtain compound of formula I as acid binding agent using inorganic bases such as cesium carbonates.

Route 2 provides a kind of illustrative methods of Formula II compound being used to prepare in route 1.

As shown in Scheme 2, Formula VII compound can be by Formula IV compound (wherein deuterated isopropylamine-d1 and deuterated isopropylamine- D7 can be bought from Aldrich) it is prepared with acetic anhydride, further, Formula VII compound and formula compound VIII are in trichlorine oxygen Production IX compound is reacted in the presence of phosphorus, organic base.Compound X can be passed through under the action of the alkali such as potassium tert-butoxide by compound IX Condensation reaction preparation.Finally, with heavy water, deuterated methanol or deuterated ethyl alcohol etc. for deuterium source, it under alkaline condition can through hydrogen-deuterium exchange Prepare Formula II compound.

Route 3 provides a kind of illustrative methods of Formula V compound being used to prepare in route 1.

As shown in Scheme 3, Formula XII compound can (can be from Aldrich by Formula XI compound and 5- bromopyridine -2- formaldehyde Purchase) it is prepared under the action of formic acid, trimethyl orthoformate.Further, compound XII can be with two silicon substrate ammonia of hexamethyl The reaction ammonification of base lithium obtains Formula V compound.

Route 4 provides the illustrative methods of another Formula V compound being used to prepare in route 1.

Formula V compound can also be prepared via route 4.Using Formula XVI Compound I as starting material, through 5- bromopyridine -2- first Aldehyde reduction amination accepted way of doing sth XXV compound, the latter can be reduced into Formula XII compound, and further ammonification is at Formula V compound

Route 5 provides the illustrative methods for the Formula V compound that another is used to prepare in route 1.

Formula V compound can also be prepared via route 5.It, (can through 6- amino-nicotinic acid using Formula XVI Compound I as starting material Bought from Aldrich) reaction accepted way of doing sth XXVI compound, the latter can be in reducing agent (such as NaBD₄(H₄)) in the presence of by further also Original is at Formula V compound

Route 6 provides another illustrative methods for being used to prepare compound of formula I.

As shown in Scheme 6, Formula XI compound and 6- amino-nicotinic acid (can buy from Aldrich) are in dehydrating agent or activating agent In the presence of preparation formula XIII compound.In the presence of a catalyst, C-N occurs for resulting Formula XIII compound and formula IV compound Coupling reaction generates intermediate Formula XIV compound, and resulting Formula XIV compound can be tried further with go back original reagent or deuterated reduction Agent such as lithium aluminium hydride reduction (LiAlH₄), deuterate aluminium lithium (LiAlD₄), sodium borohydride (NaBH₄)/lewis acid (AlCl₃,BF₃), boron deuterium Change sodium (NaBD₄)/lewis acid, borine (BH₃), deuterated borine (BD₃- THF), silane (PhSiH₃/ KOH) etc. reactions obtain Formulas I Compound.

For Formula XI compound, can be prepared by different methods for the deuterated compound of different loci.Route 7,8,9 mentions Wherein three kinds of different synthetic methods are supplied.

Route 7: the synthesis of Formula XI compound

As shown in Scheme 7, using piperazine as starting material, N, N- dinitroso piperazine is prepared through nitrozation reaction first Piperazine.It under alkaline condition, can further preparation formula XV through hydrogen-deuterium exchange with heavy water, deuterated methanol or deuterated ethyl alcohol etc. for deuterium source Compound.Under the conditions of alumel take off nitroso can preparation formula XVI compound, then with quantitative acetic anhydride, chloroacetic chloride or Acetic acidreaction is prepared into Formula XVI Compound I.Finally reducing agent for example lithium aluminium hydride reduction, deuterate aluminium lithium, sodium borohydride/lewis acid, Reaction obtains Formula XI compound under the effects of boron deuterate sodium/lewis acid.

Route 8: the synthesis of Formula XI-a compound

As shown in Scheme 8, using n-ethylpiperazine as starting material, Formula XVI IIization equally is prepared into through nitrozation reaction Close object.Under alkaline condition, with heavy water, deuterated methanol or deuterated ethyl alcohol etc. for deuterium source, Formula XI Xization is prepared into through hydrogen-deuterium exchange Close object.Nitroso is finally taken off under the conditions of alumel can be prepared into Formula XI-a compound.

Route 9: the synthesis of Formula XI-b compound

As shown in Scheme 9, it (can be bought from Aldrich) by starting material of N-benzyl piperazine, it is anti-through nitrosylation Formula X X compound should be prepared into.Under alkaline condition, it with heavy water, deuterated methanol or deuterated ethyl alcohol etc. for deuterium source, is exchanged through hydrogen-deuterium It is prepared into Formula X XI compound.Then Formula X XII compound can be prepared by nitroso being taken off under the conditions of alumel, then with it is quantitative Acetic anhydride, chloroacetic chloride or acetic acidreaction are prepared into Formula X XIII compound.Formula X XIII compound is in reducing agent such as lithium aluminium hydride reduction, deuterium Reaction obtains Formula X XIV compound under the effects of changing aluminium lithium, sodium borohydride/lewis acid, boron deuterate sodium/lewis acid.Finally with The metals such as palladium are catalyst, heating reaction debenzylation can obtain Formula XI-b compound in hydrogen environment.

Above-mentioned route, which illustrates, constitutes synthetic method of the invention, is for describing applicableization by specific embodiment Method, rather than indicate the scope of the present invention or be intended to limit.Regardless of by identical variable name (that is, R¹、R²、R³ Deng) be identified, the chemical structure in diagram depicts the chemical group definition with corresponding position in compounds herein formula herein The variable suitably limited herein.The appropriateness of chemical group exists in the chemical structural formula for synthesizing another compound Within the knowledge of those of ordinary skill in the art.

Another aspect of the invention provides a kind of pharmaceutical composition, and it includes the shown in formula I of therapeutically effective amount Compound or its pharmaceutically acceptable salt, solvate or prodrug and one or more pharmaceutically acceptable carriers or figuration Agent.

Pharmaceutical composition of the invention can be given through a variety of ways, this depend on whether locally or systemically to treat and The region treated.Can local (for example, transdermal, skin, eye and mucous membrane include, intranasal, vagina and rectum pass medicine), lung (for example, By sucking or be blown into powder or aerosol, including pass through sprayer；Intratracheally, intranasally), oral and extra-parenteral administration.Stomach External administration includes intravenous, intra-arterial, in subcutaneous, peritonaeum or intramuscular injection or infusion；Encephalic is for example intrathecal or the ventricles of the brain in give Medicine.

Certain examples of acceptable carrier or excipient include lactose, glucose, sucrose, sorbierite, mannitol, shallow lake Powder, Arabic gum, calcium phosphate, alginates, bassora gum, gelatin, calcium silicates, microcrystalline cellulose, polyvinylpyrrolidone, fiber Element, water, syrup and methylcellulose etc..Pharmaceutical composition can also contain: lubricant such as talcum powder, magnesium stearate and mineral Oil；Wetting agent；Emulsifier and suspending agent；Preservative such as methyl benzoate and benzoic acid hydroxy propyl ester；Sweetener and corrigent. Pharmaceutical composition of the invention can be prepared by using method as known in the art, so as to after giving patient provide quick-release, The effect of sustained release or delayed release of active elements.

Can be by unit dosage forms compositions formulated, every dose contains about 5~1000mg, more typically from about 100~500mg activity at Point.Term " unit dosage forms " refers to physically separated suitable as the single dose list for being used for human patient and other mammals Position, constituent parts contain mixed with the drug excipient for being suitable for be computed the active material that can produce the predetermined amount of required curative effect.

The range of the effective dose of reactive compound can be very big, is usually administered by medicinal effective quantity.However, it is to be understood that real The amount for the compound that border is given usually determines by doctor according to correlation circumstance, they include treated illness, it is selected to Medicine approach, practical compound to be administered；Age, weight and the reaction of individual patients；The severity etc. of patient symptom.

The therapeutic dose of the compounds of this invention can be depending on for example below: the particular use for the treatment of gives compound Mode, the health of patient and state, and the judgement of label prescriber.Ratio of the compounds of this invention in Pharmaceutical composition or Concentration can be not fixed, and depend on many factors, they include dosage, chemical characteristic (such as hydrophobicity) and administration route.Such as The compounds of this invention can be provided by the physiological buffer aqueous solution of the compound containing about 0.1~10%w/v, be given for parenteral Medicine.Certain exemplary dosage ranges are about 1 μ g/kg~about 1g/kg body weight/day.In certain embodiments, dosage range is about 0.01mg/kg~about 100mg/kg body weight/day.Dosage will likely depend on this class variable, such as disease or the type and hair of illness It exhibition degree, the general health status of specific patient, the relative biological efficacy of selected compound, excipient preparation and its gives Medicine approach.It can be extrapolated by the dose-response curve as derived from external or animal model test system, obtain effective dose.

Another aspect of the present invention provides compound or its pharmaceutically acceptable salt shown in formula I, solvate or preceding Purposes of the medicine in the drug of preparation treating cancer.Preferably, the cancer is selected from colorectal cancer, breast cancer, lung cancer (especially NSCLC), prostate cancer, glioblastoma, lymphoma mantle cell, chronic myelocytic leukemia and the white blood of acute myelogenous Disease.

Another aspect of the invention provides a kind of method for treating cancer in mammal, and the cancer is selected from colorectum Cancer, breast cancer, lung cancer (especially NSCLC), prostate cancer, glioblastoma, lymphoma mantle cell, the white blood of chronic granulocyte Disease and acute myeloblastic leukemia, this method include to the shown in formula I of mammal in need application therapeutically effective amount Compound or its pharmaceutically acceptable salt, solvate or prodrug.

The embodiment of the present invention compound has significant CDK4 and CDK6 inhibitory activity.For example, as described herein one In kind or many measure, there is the CDK4 inhibitory activity less than 50nM, preferably have less than the CDK4 inhibitory activity of 20nM, it is more excellent The CDK4 inhibitor activity having less than 8nM is selected, most preferably there is the CDK4 inhibitory activity less than 2nM.For example, described herein One or more measurements in, have less than 50nM CDK6 inhibitory activity, preferably have less than the CDK6 inhibitory activity of 20nM, The CDK6 inhibitor activity of more preferably less than 8nM most preferably has the CDK6 inhibitory activity less than 2nM.

Compared with Abemaciclib, certain compounds of the invention (especially embodiment compound) are in people's hepatomicrosome In there is more preferably stability and superior pharmacokinetic property, be expected to reduce clinical dosage, to reduce Treatment cost is to allow more patients to be benefited.

Specific embodiment:

The present invention can be described in more detail in the following examples, but the invention is not limited in any way.

The preparation of 1 compound I-1 of embodiment

Step 1) N- isopropyl-acetamide

2- isopropylamine (20g, 338.4mmol), triethylamine (34.6g, 341.8mmol) are successively dissolved in dichloromethane under ice bath Acetic anhydride (35.2g, 345.1mmol) is slowly dropped in reaction flask, in control by alkane (150mL) by constant pressure funnel Temperature is less than 5 DEG C.It finishes, reaction flask is transferred to and is stirred overnight at room temperature.Reduced pressure removes solvent, uses methyl tertiary butyl ether(MTBE) Potassium carbonate (50g) is added after (200mL) dilution, stirs 1 hour at room temperature.Decompression filters, and filtrate decompression is concentrated to give the change of Formula VII -1 It closes object (27.8g), is weak yellow liquid.

¹H-NMR(CDCl₃, 300MHz): δ 5.71 (s, 1H), 4.06 (dt, 1H), 1.94 (s, 3H), 1.14 (d, J= 6.5Hz,6H)。

HRMS (M+H) m/z:102.0908.

Step 2) N- (the bromo- 2,6- difluorophenyl of 4-)-N '-isopropyl-second is narrowed

By bromo- 2, the 6- difluoroaniline (31.05g, 0.15mol) of 4-, N- isopropyl yl acetamide (30.3g, 0.30mmol), three Chlorethoxyfos (20.9mL, 0.225mol) are added sequentially in dry toluene.Under ice bath by triethylamine (31.3mL, 0.225mol) in It is slowly dropped to reaction flask in constant pressure funnel, keeps interior temperature less than 60 DEG C.Reaction flask, which is transferred in oil bath pan, is heated to solvent time Stream.Reaction flask is cooled to room temperature after 2 hours, is poured slowly into 300g mixture of ice and water, and 300 milliliters of ethyl acetate are added, fill Divide liquid separation after mixing, water layer uses 200 milliliters of ethyl acetate extractions again, merges organic layer, anhydrous sulphur after saturated common salt water washing Sour sodium is dry, and faint yellow solid is obtained after reduced pressure, and 100 milliliters of petroleum ethers are added and are beaten 10 minutes, decompression filters to obtain Formula IX -1 Compound (28.0g), for off-white powder (yield 92.3%).

¹H-NMR(CDCl₃, 300MHz): δ 7.01~7.03 (m, 2H), 4.17 (s, 1H), 1.76 (s, 3H), 1.23 (d, J =5.6Hz, 6H).

¹³C-NMR (DMSO, 300MHz): δ 157.48,156.65,153.41,115.09,114.72,110.03,41.62, 21.79,17.62。

HRMS (M+H) m/z:291.0088.

The fluoro- 1- isopropyl -2- methyl-1 H- benzimidazole of the bromo- 4- of step 3) 6-

N- (bromo- 2, the 6- difluorophenyl of 4-)-N '-isopropyl-second is narrowed (27.2g, 93.47mmol) and is dissolved in anhydrous N, N- bis- Methylformamide (200mL) is added potassium tert-butoxide (13.11g, 116.83mmol), is heated to 110 DEG C under nitrogen protection.4 hours Reaction flask is cooled to room temperature afterwards, and 500 milliliters of ethyl acetate are added, and 800 milliliters of water mix well rear liquid separation, and water layer continues with 500 Milliliter ethyl acetate extraction, liquid separation.Merge organic layer, first with 200 milliliters water washing 5 times, then with the washings of 200 milliliters of saturated common salts It washs 3 times, Light brown solid is concentrated under reduced pressure to obtain after organic layer anhydrous sodium sulfate is dry, after 100 milliliters of petroleum ether mashing twice are added - 1 compound of Formula II (21.0g) is obtained, is faint yellow solid.

¹H-NMR(CDCl₃, 300MHz): δ 7.41 (d, J=1.2Hz, 1H), 7.07 (dd, J₁=1.4Hz, J₂=9.6Hz, 1H), 4.62 (heptet, 1H), 2.62 (s, 3H), 1.62 (d, J=7.0Hz, 6H).

¹³C-NMR(CDCl₃, 300MHz): δ 154.75,151.91,151.35,137.11,130.90,111.14, 110.27,48.39,21.17,14.79。

HRMS (M+H) m/z:271.0251.

The fluoro- 1- isopropyl -2- [D of the bromo- 4- of step 4) 6-₃] methyl-1 H-[5,7-D₂] benzimidazole

Heavy water (15mL), the heavy aqueous solution (2mL) of 40% content NaOD, anhydrous tetrahydro furan (5mL) are added sequentially to In microwave reaction bottle, the fluoro- 1- isopropyl -2- methyl-1 H- benzimidazole (2.76g, 10mmoL) of the bromo- 4- of 6- is then added.It opens Microwave heating reaction, setting reaction temperature are 145 DEG C, and the reaction time is 15 hours.End of reaction, product are sunken to reaction flask bottom Heavy aqueous solution (2mL), the anhydrous tetrahydro furan of heavy water (15mL), 40% content NaOD are added again after removing solvent in portion (5mL) continues microwave reaction under same reaction conditions (145 DEG C of reaction temperature, 15 hours reaction time), repeats deuterated 4 times.It can Obtain -2 compound of Formula II (2.30g).

¹H-NMR (DMSO, 300MHz): δ 4.76 (heptet, 1H), 1.54 (d, J=6.9Hz, 6H).

MS (M+H): 276.0553.

¹³C-NMR (DMSO, 300MHz): δ 153.91,152.79,150.55,137.34,130.51,112.48, 110.68,47.87,20.63。

HRMS (M+H) m/z:276.0557.

The fluoro- 1- isopropyl -2- [D of step 5) 4-₃] methyl -6- (4,4,5,5- tetramethyl-[1,3,2] dioxa boron heterocycle Pentane -2- base) -1H- [5,7-D₂] benzimidazole

DMSO (100mL) is added in 500 milliliters of single-necked flasks, the fluoro- 1- isopropyl -2- of the bromo- 4- of 6- is sequentially added [D₃] methyl-1 H- [5,7-D₂] benzimidazole (20.7g, 73.80mmoL), bis- (pinacol) two boron (27.6g, 108.69mmol), tricyclohexyl phosphine (3.53g, 12.61mmol), potassium acetate (21.3g, 217.38mmol).Nitrogen displacement reaction It is rapidly joined acid chloride (1.5g) after air in bottle, is opened under nitrogen protection and be heated to 90 DEG C.Reaction flask is cooled to room after 3 hours Temperature, reaction solution pour into 700 milliliters of water, depressurize and filter after being sufficiently mixed, and filter cake is lightly dry afterwards twice with 100 milliliters of water washings Brown solid.50mL petroleum ether is added in crude product, and 10mL ethyl acetate is beaten 10min, and decompression filters to obtain -1 compound of formula III (18.8g) is off-white powder.

¹H-NMR (DMSO, 300MHz): δ 4.82 (heptet, 1H), 1.56 (d, J=6.9Hz, 6H), 1.32 (s, 12H).

¹³C-NMR (DMSO, 300MHz): δ 154.08,153.12,150.76,136.62,133.79,113.61, 110.99,83.74,47.68,24.59,22.90,21.00。

HRMS (M+H) m/z:324.2311.

Step 6) 6- (the chloro- 5-FU -4- base of 2-) fluoro- 1- isopropyl -2- [D of -4-₃] methyl-1 H- [5,7-D₂] benzo Imidazoles

2,4-, bis- chloro- 5-FU (9.95g, 59.60mmol) is dissolved in glycol dimethyl ether (210mL), is sequentially added The fluoro- 1- isopropyl -2- [D of 2M aqueous sodium carbonate (140mL), 4-₃] methyl -6- (4,4,5,5- tetramethyl-[1,3,2] dioxy Miscellaneous boron heterocycle pentane -2- base) -1H- [5,7-D₂] benzimidazole (17.5g, 54.18mmol), bis- (triphenylphosphine) palladium chlorides (1.0g).Air in nitrogen displacement reaction flask opens oil bath heating, is stirred to react 1h at 85 DEG C.Reaction flask is cooled to room temperature, instead Liquid is answered to pour into 300mL water, a large amount of solids are precipitated, and depressurize and filter after being sufficiently stirred, and -1 compound of formula IV is obtained after filtration cakes torrefaction (13.18g) is pale solid.

¹H-NMR (DMSO, 300MHz): δ 8.94 (d, J=3.4Hz, 1H), 4.86 (heptet, 1H), 1.60 (d, J= 6.9Hz,6H)。

¹³C-NMR(CDCl₃, 300MHz): δ 156.45,154.81,153.85,152.95,150.54,149.58, 149.22,136.35,124.50,109.15,106.98,47.93,20.77,14.50(m,CD₃)。

HRMS (M+H) m/z:328.1184.

Step 7) 1- (the bromo- pyridin-3-yl methyl of 6-) -4- ethyl-piperazin

N-ethylpiperazine (1.59g, 13.95mmol), the bromo- 5- pyridine carboxaldehyde (3.14g, 16.88mmol) of 2- are successively added Enter into acetonitrile, formic acid (2.10mL, 55.80mmol), trimethyl orthoformate (3.07mL, 27.90mmol) is added.Nitrogen protection Under be heated to reflux, reaction solution is cooled to room temperature after 4 hours, be added 30mL water, 15mL ethyl acetate, liquid separation.Organic layer is residue The bromo- 5- pyridine carboxaldehyde of 2-.Water layer is added saturation sodium hydroxide and adjusts pH to 10,30mL ethyl acetate is added, liquid separation, water layer is again 30mL ethyl acetate is added, merges organic layer, product crude product is concentrated under reduced pressure to obtain after anhydrous sodium sulfate is dry.Column chromatographic purifying obtains formula XII-1 compound (1.48g) is colourless liquid.

¹H-NMR(300MHz,CDCl₃): δ 8.29 (1H, d, J=1.74Hz, Py), 7.53-7.55 (1H, dd, J= 1.98Hz, 8.10Hz, Py), 7.43 (1H, d, J=8.10Hz, Py), 3.47 (2H, s, CH₂),2.38-2.48(10H,m, piper-CH₃ CH₂ ), 1.08 (3H, t, J=7.14Hz,CH₃ CH₂)。

¹³C-NMR(300MHz,CDCl₃): δ 150.59,140.71,139.29,133.18,127.72,59.25,52.61, 52.21,11.84。

HRMS (M+H) m/z:284.0757.

Step 8) 5- (4- ethyl-piperazin -1- ylmethyl)-pyridine -2- base amine

1- (the bromo- pyridin-3-yl methyl of 6-) -4- ethyl-piperazin (960mg, 3.38mmol) is dissolved in anhydrous tetrahydro furan (8mL), be added 2- (dicyclohexyl phosphino-) biphenyl (120mg, 0.338mmol), tris(dibenzylideneacetone) dipalladium (154mg, 0.169mmol).Lithium hexamethyldisilazide (4.06mL, 1M, 4.06mmol) is slowly added under nitrogen protection.Reaction flask oil Bath is heated to 65 DEG C, and reaction flask is cooled to room temperature after twenty minutes, and 50mL ethyl acetate, 30mL water, liquid separation after mixing, water layer is added 30mL ethyl acetate is added again, liquid separation merges organic layer, rotates to obtain Light brown solid after anhydrous sodium sulfate is dry.Column chromatography - 1 compound of Formula V (670mg) is purified to obtain, is faint yellow solid.

¹H-NMR(300MHz,CDCl₃): δ 7.75 (1H, d, J=1.65Hz, Py), 7.24-7.28 (1H, dd, J= 2.16Hz, 8.37Hz, Py), 6.40 (1H, d, J=8.37Hz, Py), 5.77 (2H, s, NH₂),3.24(2H,s,CH₂),2.25- 2.32(10H,m,piper-CH₃ CH₂ ), 0.96 (3H, t, J=7.11Hz,CH₃ CH₂)。

¹³C-NMR(300MHz,CDCl₃): δ 157.64,148.54,139.12,123.18,108.27,59.74,52.64, 52.20,11.84。

HRMS (ESI) m/z:220.1688.

Step 9) [5- (4- ethyl piperazidine -1- ylmethyl)-pyridine -2- base]-[5 fluoro- 4- (the fluoro- 3- isopropyl -2- of 7- [D₃] methyl -3H- [5,7-D₂] -5 base of benzimidazole)-pyrimidine -2-base]-amine

By 6- (the chloro- 5-FU -4- base of 2-) the fluoro- 1- isopropyl -2- [D of -4-₃] methyl-1 H- [5,7-D₂] benzimidazole (788mg, 2.41mmol), 5- (4- ethyl piperazidine -1- ylmethyl)-pyridine -2- base amine (530mg, 2.41mmol), cesium carbonate (1.57g, 4.82mmol) is added sequentially in dioxane (20mL), and air in nitrogen displacement reaction flask rapidly joins three (two BENZYLIDENE ACETONE) two palladiums (50mg), the double diphenylphosphine xanthenes (80mg) of 9,9- dimethyl -4,5-.It is heated under nitrogen protection Reaction solution reflux.Reaction solution is cooled to room temperature after 2 hours, and 30mL ethyl acetate is added, and decompression filters.20mL water is added in filtrate, Liquid separation, when water layer, enter 20mL ethyl acetate, and liquid separation merges organic layer, are concentrated under reduced pressure after anhydrous sodium sulfate is dry and remove solvent and obtain Light brown solid.9mL petroleum ether is added in crude product, and 3mL ethyl acetate is stirred at room temperature 30 minutes, and decompression filters, and obtains -1 chemical combination of Formulas I Object 858mg, for off-white powder (yield 69.8%).

¹H-NMR (DMSO, 300MHz): δ 10.08 (s, 1H), 8.67 (s, 1H), 8.20~8.22 (m, 1H), 7.65 (d, J =7.7Hz, 1H), 4.81~4.85 (m, 1H), 3.42 (s, 2H), 2.27~2.36 (m, 10H), 1.62 (d, J=6.4Hz, 6H), 0.96 (t, J=6.6Hz, 3H).

¹³C-NMR (DMSO, 300MHz): δ 155.34,154.46,153.85,152.11,150.55,148.73, 148.07,147.77,147.41,138.17,136.36,133.38,126.97,126.40,111.69,108.81,58.77, 52.44,51.52,48.03,20.88,11.96 (deuterated methyl carbon multiplet packet is in noise).

HRMS (M+H) m/z:512.3103.

The preparation of 2 compound I-10 of embodiment

The fluoro- 1- isopropyl -2- methyl -6- of step 1) 4- (4,4,5,5- tetramethyl-[1,3,2] dioxaborolan alkane - 2- yl) -1H- benzimidazole

The fluoro- 1- isopropyl -2- [D of the bromo- 4- of 6- is replaced with the fluoro- 1- isopropyl -2- methyl-1 H- benzimidazole of the bromo- 4- of 6-₃] Methyl-1 H-[5,7-D₂] benzimidazole, and the method referring to shown in 1 step 5) of embodiment prepares preparation formula III-2 compound.

¹H-NMR(CDCl₃, 300MHz): δ 7.69 (s, 1H), 7.33 (d, J=10.7Hz, 1H), 4.65~4.74 (m, 1H), 2.65 (s, 3H), 1.65 (d, J=7.0Hz, 6H), 1.36 (s, 12H).

¹³C-NMR(CDCl₃, 300MHz): δ 154.89,152.20,151.55,136.67,134.27,113.46, 112.51,83.88,48.28,24.81,21.43,15.12。

HRMS (M+H) m/z:319.1987.

The fluoro- 1- isopropyl -2- methyl-1 H- benzimidazole of step 2) 6- (the chloro- 5-FU -4- base of 2-) -4-

With the fluoro- 1- isopropyl -2- methyl -6- of 4- (4,4,5,5- tetramethyl-[1,3,2] dioxaborolan alkane -2- Base) -1H- benzimidazole replace the fluoro- 1- isopropyl -2- [D of 4-₃] methyl -6- (4,4,5,5- tetramethyl-[1,3,2] dioxa boron Heterocycle pentane -2- base) -1H- [5,7-D₂] benzimidazole, and the method preparation formula IV-2 chemical combination referring to shown in 1 step 6) of embodiment Object.

¹H-NMR (DMSO, 300MHz): δ 8.95 (d, J=3.3Hz, 1H), 8.15 (s, 1H), 7.62 (d, J=11.9Hz, 1H), 4.86 (heptet, 1H), 2.65 (s, 3H), 1.60 (d, J=6.9Hz, 6H).

¹³C-NMR (DMSO, 300MHz): δ 156.60,155.03,153.96,153.10,150.66,149.78, 149.42,136.49,124.66,109.36,107.08,48.05,20.88,14.63。

HRMS (M+H) m/z:323.0870.

Step 3) N- benzyl-N- nitroso piperazine

N-benzyl piperazine (50.0g, 0.284mol) is added into reaction flask, 0 DEG C or so dropwise addition 2M HCl (160ml, 0.318mol), the reaction solution obtained stirs 10 minutes at 0 DEG C, and NaNO is then added dropwise into reaction₂Aqueous solution [NaNO₂, The solution that water (58mL) is made into is added in (24.50g, 0.335mol)], it maintains temperature at 0~10 DEG C, drips off within 40 minutes.Reaction is mixed Object is closed to be stirred at room temperature overnight.Reaction is heated to 35 DEG C about 1 hour (a large amount of insoluble solids generations) by next day, then by reaction flask It is put into cryostat, it is cooling at 0 DEG C to be precipitated, solid product is isolated finally by suction filtration, is washed, water layer is extracted with DCM again, had Machine mutually merges, and anhydrous magnesium sulfate is dry, removes desiccant by filtering, rotary evaporation removes solvent and obtains solid product.Finally By obtaining Formula X X-1 compound (56.71g) after decompression 50 DEG C of dryings of baking oven, it is faint yellow solid (yield 97.30%).

¹H-NMR(300MHz,CDCl₃): 7.26~7.34 (m, 5H), 4.23~4.27 (t, J=10.4Hz, 2H), 3.81 ~3.85 (t, J=10.7Hz, 2H), 3.60 (s, 2H), 2.66~2.70 (t, J=10.3Hz, 2H), 2.43~2.46 (t, J= 10.7Hz,2H)。

MS (M+H) m/z:206.1.

Step 4) N- benzyl-N- nitroso-[3,3,5,5-D₄] piperazine

N- benzyl-N- nitroso piperazine (20.0g, 97.43mmol) is added into reaction flask, sodium methoxide (15.79g, 292.29mmol), under nitrogen, heavy water (150ml) slowly is added into reaction flask, deuterated ethyl alcohol (d1,100ml).Then it heats To 80 DEG C, dissolved clarification is reacted.Stop heating after 24 hours, it is to be cooled to be put into cryostat, to after room temperature, it is cooling at 0 DEG C to be precipitated.3 After hour, solid product is isolated by filtering.Formula X XI-1 compound is dried to obtain for 50 DEG C of baking oven finally by decompression (17.735g) is faint yellow solid.

¹H-NMR(300MHz,CDCl₃): 7.25~7.33 (m, 5H), 3.56 (s, 2H), 2.64 (s, 2H), 2.41 (s, 2H)。

MS ESI, (M+Na) m/z:232.2.

Step 5) N- benzyl-[3,3,5,5-D₄] piperazine

Formula X XI-1 compound (10.0g, 47.78mmol) is added into reaction flask, sodium methoxide (7.74g, 143.34mmol), under nitrogen, heavy water (75mL) and deuterated ethyl alcohol (d1,75mL) are slowly added into reaction.It is then heated to 70 ℃.Stop heating after reaction 24 hours, after being cooled to room temperature, Al-Ni alloy (30.0g) is added into reaction several times, about 2 Hour adds, and reaction mixture is stirred at room temperature overnight.Next day removes solid metal by filtering, and collection contains product Filtrate.It is extracted again with DCM, organic phase merges, and anhydrous magnesium sulfate is dry, removes desiccant by filtering, rotary evaporation removes molten Agent obtains solid product.It is that faint yellow oily body (produces finally by Formula X XII-1 compound (7.13g) is obtained after being dried under reduced pressure Rate 82.81%).

¹H-NMR(300MHz,CD₃OD): 7.20~7.31 (m, 5H), 3.47 (s, 2H), 2.39 (s, 4H).

HRMS (ESI, M+H) m/z:181.2.

Step 6) N- benzyl-N '-acetyl group-[3,3,5,5-D₄] piperazine

By N- benzyl-[3,3,5,5-D₄] piperazine (5.40g, 30mmol) is dissolved in methylene chloride (30mL), under condition of ice bath It is slowly added to acetic anhydride (3.4mL, 36mmol), is stirred at room temperature 10 minutes, 20mL saturated sodium carbonate solution is added in reaction solution, point 30mL methylene chloride is added in liquid, water layer, and organic phase is merged after liquid separation, Formula X XIII-1 is concentrated under reduced pressure to obtain after anhydrous sodium sulfate is dry Compound (6.20g) is light brown liquid.

Step 7) N- benzyl-N '-[1,1-D₂] ethyl-[3,3,5,5-D₄] piperazine

By N- benzyl-N '-acetyl group-[3,3,5,5-D₄] piperazine (3.33g, 15.0mmol) is dissolved in anhydrous tetrahydro furan (30mL), sequentially adds [D4] boron deuterate sodium (1.23g, 30mmol), aluminium chloride (4.01g, 30mmol), and unlatching is heated to reacting Liquid reflux.Reaction solution is cooled to room temperature after 3 hours, is poured into 50g mixture of ice and water, and saturation sodium hydroxide solution is added and adjusts pH To 10 or so.50mL ethyl acetate, liquid separation after mixing is added, 50mL ethyl acetate is added in water layer again, and liquid separation merges organic Formula X XIV-1 compound (2.85g) is concentrated under reduced pressure to obtain after anhydrous sodium sulfate is dry in layer, is colourless liquid.

¹H-NMR (DMSO, 300MHz): δ 7.21~7.33 (m, 5H), 3.43 (s, 2H), 2.33 (s, 4H), 0.95 (s, 3H)。

¹³C-NMR (DMSO, 300MHz): δ 138.27,128.67,128.02,126.74,69.66,62.13,52.47, 11.70。

HRMS (M+H) m/z:211.1454.

Step 8) N- [1,1-D₂] ethyl-[2,2,6,6-D₄] piperazine

By N- benzyl-N '-[1,1-D₂] ethyl-[3,3,5,5-D₄] piperazine (2.75g, 13.10mmol) is dissolved in methanol, add Enter 10% content palladium carbon (0.28g).60 DEG C are heated in hydrogen environment.Reaction solution decompression filters after 10 hours, and filtrate rotates - 2 compound of Formula XI (1.80g) is colourless liquid.

¹H-NMR (DMSO, 300MHz): δ 2.74 (s, 4H), 0.95 (s, 3H).

¹³C-NMR (DMSO, 300MHz): δ 69.76,52.59,44.56,26.06,11.70.

HRMS (M+H) m/z:121.1282.

Step 9) 1- (the bromo- pyridin-3-yl methyl of 6-) -4- [1,1-D₂] ethyl-[3,3,5,5--D₄] piperazine

With N- [1,1-D₂] ethyl-[2,2,6,6-D₄] piperazine XI-2 replaces n-ethylpiperazine, and referring to 1 step of embodiment 7) the XII-2 compound of method preparation formula shown in.

¹H-NMR (DMSO, 300MHz): δ 8.29 (s, 1H), 7.65~7.68 (m, 1H), 7.58~7.61 (m, 1H), 3.46(s,2H),2.36(s,4H),0.96(s,3H)。

¹³C-NMR (DMSO, 300MHz): δ 150.51,139.94,139.77,133.67,127.62,69.66,58.08, 52.09,11.51。

MS (M+H) m/z:290.1127.

Step 10) 5- (4- [1,1-D₂] ethyl-[3,3,5,5-D₄] piperazine -1- ylmethyl)-pyridine -2- base amine

With 1- (the bromo- pyridin-3-yl methyl of 6-) -4- [1,1-D₂] ethyl-[3,3,5,5-D₄] piperazine replace 1- (the bromo- pyrrole of 6- Pyridine -3- ylmethyl) -4- ethyl-piperazin, and the method preparation formula V-2 compound referring to shown in 1 step 8) of embodiment.

¹H-NMR (DMSO, 300MHz): δ 7.75 (d, J=1.6Hz, 1H), 7.26 (dd, J₁=2.1Hz, J₁=8.4Hz 1H), 6.39 (d, J=8.4Hz, 1H), 5.77 (s, 2H), 3.23 (s, 2H), 2.29 (s, 4H), 0.94 (s, 3H).

¹³C-NMR (DMSO, 300MHz): δ 158.93,147.95,138.05,120.78,107.50,59.14,52.14, 11.70。

HRMS (M+H) m/z:227.2136.

Step 11) [5- (4- [1,1-D₂] ethyl-[3,3,5,5-D₄] piperazine -1- ylmethyl)-pyridine -2- base]-[5 is fluoro- 4- (fluoro- -5 base of 3- isopropyl -2- methyl -3H- benzimidazole of 7-)-pyrimidine -2-base]-amine

6-- (2- is replaced with the fluoro- 1- isopropyl -2- methyl-1 H- benzimidazole of 6- (the chloro- 5-FU -4- base of 2-) -4- Chloro- 5-FU -4- base) the fluoro- 1- isopropyl -2- [D of -4-₃] methyl-1 H- [5,7-D₂] benzimidazole, with 5- (4- [1,1-D₂] Ethyl-[3,3,5,5-D₄] piperazine -1- ylmethyl) and-pyridine -2- base amine replace 5- (4- ethyl piperazidine -1- ylmethyl)-pyridine - 2- base amine, and the method preparation formula I-10 compound referring to shown in 1 step 9 of embodiment.

¹H-NMR (DMSO, 300MHz): δ 10.12 (s, 1H), 8.69 (d, J=2.9Hz, 1H), 8.29 (s, 1H), 8.20 ~8.22 (m, 2H), 7.66~7.70 (m, 2H), 4.80~4.89 (m, 1H), 3.43 (s, 2H), 2.64 (s, 3H), 2.36 (s, 4H), 1.63 (d, J=6.5Hz, 6H), 0.95 (s, 3H).

¹³C-NMR (DMSO, 300MHz): δ 155.33,154.52,153.89,152.12,150.59,148.75, 148.09,147.79,147.43,138.19,136.37,133.36,127.00,126.56,111.72,108.88,58.81, 52.29,51.45,48.05,20.89,14.50,11.72。

HRMS (M+H) m/z:513.3152.

The preparation of 3 compound I-9 of embodiment

Step 1) N- benzyl-N '-ethyl-[3,3,5,5-D₄] piperazine

Boron deuterate sodium, and the method preparation formula XXIV-2 compound referring to shown in 2 step 7) of embodiment are replaced with lithium aluminium hydride.

¹H-NMR (DMSO, 300MHz): δ 7.20~7.33 (m, 5H), 3.43 (s, 2H), 2.26~2.34 (m, 6H), 0.97 (t, J=7.2Hz, 3H).

¹³C-NMR (DMSO, 300MHz): δ 138.25,128.70,128.04,126.77,62.10,52.41,51.44, 11.92。

MS (M+H): 209.1346.

Step 2) N- ethyl-[2,2,6,6-D₄] piperazine

With N- benzyl-N '-ethyl-[3,3,5,5-D₄] piperazine replace N- benzyl-N '-[1,1-D₂] ethyl-[3,3,5,5- D₄] piperazine, and the method preparation formula XI-3 compound referring to shown in 2 step 8 of embodiment.

¹H-NMR (DMSO, 300MHz): δ 2.73 (s, 4H), 2.28 (q, J=7.2Hz, 2H), 0.97 (t, J=7.2Hz, 3H)。

¹³C-NMR (DMSO, 300MHz): δ 169.76,51.96,44.73,26.06,11.71.

HRMS (M+H) m/z:119.1475.

Step 3) 1- (the bromo- pyridin-3-yl methyl of 6-) -4- ethyl-[3,3,5,5-D₄] piperazine

With N- ethyl-[2,2,6,6-D₄] piperazine replace N- [1,1-D₂] ethyl-[2,2,6,6-D₄] piperazine, and referring to reality Apply method preparation formula XII-3 compound shown in 2 step 9) of example.

¹H-NMR (DMSO, 300MHz): δ 8.28 (1H, s, Py), 7.64-7.68 (1H, m, Py), 7.59 (1H, d, J= 8.10Hz,Py),3.46(2H,s,CH₂-Py),2.35(4H,s,CD2CH₂- piper), 2.28 (2H, q, J=7.20Hz, CH₃CH₂- piper), 0.96 (3H, t, J=7.20Hz, CH₃CH₂)。

¹³C-NMR (DMSO, 300MHz): δ 150.47,139.89,139.73,133.70,127.59,58.12,52.28, 51.37,26.02,11.91。

HRMS (M+H) m/z:288.0843,290.0825.

Step 4) 5- (4- ethyl-[3,3,5,5-D₄] piperazine -1- ylmethyl)-pyridine -2- base amine

With 1- (the bromo- pyridin-3-yl methyl of 6-) -4- ethyl-[3,3,5,5-D₄] piperazine replace 1- (the bromo- pyridin-3-yl of 6- Methyl) -4- [1,1-D₂] ethyl-[3,3,5,5-D₄] piperazine, and the method preparation formula V-3 chemical combination referring to shown in 2 step 10 of embodiment Object.

¹H-NMR (DMSO, 300MHz): δ 7.74 (1H, s, Py), 7.25 (1H, d, J=8.34HzPy), 6.38 (1H, d, J =8.37Hz, Py), 3.23 (2H, s, CH₂-Py),2.30(4H,s,CD2CH₂- piper), 2.26 (2H, q, J=7.20Hz, CH₃CH₂- piper), 0.96 (3H, t, J=7.14Hz, CH₃CH₂)。

¹³C-NMR (DMSO, 300MHz): δ 158.90,147.93,138.03,120.81,107.48,59.13,52.16, 51.41,11.95。

HRMS (M+H) m/z:225.1881.

Step 5) [5- (4- ethyl-[3,3,5,5-D₄] piperazine -1- ylmethyl)-pyridine -2- base]-[(7- is fluoro- by 5 fluoro- 4- - 5 base of 3- isopropyl -2- methyl -3H- benzimidazole)-pyrimidine -2-base]-amine

With 5- (4- ethyl-[3,3,5,5-D₄] piperazine -1- ylmethyl) and-pyridine -2- base amine replace 5- (4- [1,1-D₂] second Base-[3,3,5,5-D₄] piperazine -1- ylmethyl)-pyridine -2- base amine, and the method preparation formula referring to shown in 2 step 11) of embodiment I-9 compound.

¹H-NMR (DMSO, 300MHz): δ 10.08 (s, 1H), 8.68 (d, J=2.6Hz, 1H), 8.29 (s, 1H), 8.20 ~8.22 (m, 2H), 7.65~7.70 (m, 2H), 4.80~4.89 (m, 1H), 3.45 (s, 2H), 2.64 (s, 3H), 2.35~ 2.55 (m, 6H), 1.63 (d, J=6.4Hz, 6H), 0.99 (t, J=6.7Hz, 3H).

¹³C-NMR (DMSO, 300MHz): δ 155.35,154.51,153.89,152.15,150.59,148.75, 148.13,147.76,147.41,138.22,136.38,133.35,126.82,126.47,111.71,108.87,58.65, 51.82,51.29,48.05,30.36,20.88,14.48,11.50。

HRMS(M+H)m/z:511.3017。

The preparation of 4 compound I-4 of embodiment

Step 1) N, N- dinitrosopiperazine

Piperazine (8.0g, 93.0mmol) is added into reaction flask, 0 DEG C or so dropwise addition 2M HCl (104ml, 208mmol), 0 NaNO is added dropwise after stirring 10min at DEG C₂Aqueous solution (NaNO₂, 15.2g, water 36ml), it is added dropwise within 30 minutes or so, room temperature is anti- It should stay overnight.It is heated to 35 DEG C to react about 1-2 hours, 0 DEG C of cooling precipitation filters to obtain faint yellow solid, 45 DEG C of vacuum oven bakings N, N- dinitrosopiperazine (12.32g), yield 92% are obtained after dry.

¹H-NMR(400MHz,CDCl₃): δ 4.60 (s, 1.7H), 4.46 (t, J=5.4Hz, 2.3H), 4.21 (t, 2.3H), 3.88 (s, 1.7H)-(form inseparable two isomers ,-N-N=O by nitroso Liang Zhong different orientation on nitrogen).

¹³C-NMR(400MHz,CDCl₃): δ 49.58,47.09,40.50,37.75 (inseparable two isomers ,- N-N=O)

HRMS (ESI) m/z:144.0647.

Step 2) N, N- dinitroso-[D₈] piperazine

Dinitrosopiperazine (10.0g, 69.4mmol) is added into reaction flask, sodium methoxide (15.0g, 277.7mmol), nitrogen It is slowly added under gas heavy water (300mL, 15.0mol), 80 DEG C are reacted 10 hours.It is cooling at 0 DEG C to be precipitated, it filters, vacuum oven 45 DEG C dry to obtain -1 compound of Formula XV (8.56g), are faint yellow solid, yield 82%.

¹H-NMR(300MHz,CDCl₃) (paranitroanisole does internal standard): δ 8.20 (2H, d, J=9.21Hz, Ph), 6.96 (2H, d, J=9.21Hz, Ph), 3.91 (3H, s, CH₃) (in addition to internal standard molecular hydrogen peak, without other hydrogen peaks, hydrogen whole quilt on piperazine Deuterium replaces).

HRMS (M+H) m/z:153.1212.

Step 3) [D₈] piperazine dihydrochloride

Dinitroso-D is added into reaction flask₈Piperazine (3.0g, 19.74mmol), sodium methoxide (5.99g, 0.11mol), It is slowly added to heavy water (30mL, 15.0mol) and the deuterated ethyl alcohol of D1- (30mL), Al-Ni alloy is added after being cooled to room temperature several times (24.0g), reaction is stayed overnight at room temperature.It filters, dense salt is slowly added dropwise after liquid all steams in filtrate air-distillation (130 DEG C) Sour (4mL).Revolving removes solvent and obtains white solid, and 45 DEG C of vacuum oven dry to obtain XVI-1 compound (2.86g), yield 82%.

¹H-NMR(300MHz,CDCl₃) (paranitroanisole does internal standard): δ 8.20 (2H, d, J=9.27Hz, Ph), 7.07 (2H, d, J=9.27Hz, Ph), 3.91 (3H, s, CH₃) (in addition to internal standard molecular hydrogen peak, without other hydrogen peaks, hydrogen whole quilt on piperazine Deuterium replaces).

HRMS (M+H) m/z:95.1417.

Step 4) N- acetyl group-[D₈] piperazine

- 1 compound of Formula XVI (0.5g, 2.99mmol) is added in single neck bottle of 25ml, water (5mL), then plus Na₂CO₃ PH to 8 or so is adjusted, acetic anhydride (0.34mL, 3.59mmol) is slowly added dropwise under ice bath, is stirred 10 minutes at room temperature.DCM extraction removes Decontamination, water phase, to alkalinity, are spin-dried for obtaining white solid, DCM mashing merges organic phase, is concentrated after filtering with the NaOH tune pH of 2M Formula XVI I-1 compound (0.23g) is obtained, is white solid, yield 56.5%.

¹H-NMR(300MHz,CDCl₃): δ 2.02 (3H, s, CH₃)。

HRMS (M+H) m/z:137.1526.

Step 5) 1- (the bromo- pyridin-3-yl methyl of 6-) -4- acetyl group-[D₈] piperazine

With N- acetyl group-[D₈] piperazine replaces n-ethylpiperazine, and the method preparation formula referring to shown in 1 step 7 of embodiment XXV-1 compound.

¹H-NMR(300MHz,CDCl₃): δ 8.29 (1H, d, J=1.65Hz, Py), 7.62-7.66 (1H, dd, J= 2.40Hz, 8.16Hz, Py), 7.43 (1H, d, J=8.10Hz, Py), 3.48 (2H, s, CH₂),2.09(3H,s,CH₃)。

HRMS (M+H) m/z:306.1057.

Step 6) 1- (the bromo- pyridin-3-yl methyl of 6-) -4- ethyl-[D₈] piperazine

Formula X XV-1 compound (3.07g, 10.03mmol) is added in the mono- neck bottle of 100ml, anhydrous THF (35mL), NaBH₄ (1.52g, 40.10mmol), is added portionwise AlCl₃(5.35g, 40.10mmol), room temperature reaction is overnight.2M hydrochloric acid is added to molten Clearly, 70 DEG C of reflux 1h, 2M sodium hydroxide solution tune pH are extracted to alkalinity, DCM, and anhydrous sodium sulfate is dry.Column chromatographic isolation and purification (DCM:MeOH=5:1) -4 compound of Formula XII (1.91g) is obtained, is yellow liquid, yield 72%.

¹H-NMR(300MHz,CDCl₃): δ 8.29 (1H, d, J=1.74Hz, Py), 7.62-7.66 (1H, dd, J= 2.40Hz, 8.10Hz, Py), 7.42 (1H, d, J=8.10Hz, Py), 3.48 (2H, s, CH₂), 2.40-2.47 (2H, q, J= 7.23Hz,CH₃ CH₂ ), 1.09 (3H, t, J=7.23Hz,CH₃ CH₂)。

HRMS (M+H) m/z:292.1245.

Step 7) 5- (4- ethyl-[D₈] piperazine -1- ylmethyl)-pyridine -2- amine

With 1- (the bromo- pyridin-3-yl methyl of 6-) -4- ethyl-[D₈] piperazine replace 1- (the bromo- pyridin-3-yl methyl of 6-) -4- Ethyl-piperazin, and the method preparation formula V-4 compound referring to shown in 1 step 8) of embodiment.

¹H-NMR(300MHz,CDCl₃): δ 7.95 (1H, d, J=1.65Hz, Py), 7.39-7.43 (1H, dd, J= 2.04Hz, 8.34Hz, Py), 6.47 (1H, d, J=8.34Hz, Py), 4.44 (2H, s, NH₂),3.37(2H,s,CH₂),2.37- 2.45 (2H, q, J=7.20Hz, CH₃ CH₂ ), 1.07 (3H, t, J=7.20Hz,CH₃ CH₂)。

HRMS (M+H) m/z:229.2268.

Step 8) [5- (4- ethyl-[D₈] piperazine -1- ylmethyl)-pyridine -2- base]-[5 fluoro- 4- (the fluoro- 3- isopropyls-of 7- - 5 base of 2- methyl -3H- benzimidazole)-pyrimidine -2-base]-amine

Replacing 6- with the fluoro- 1- isopropyl -2- methyl-1 H- benzimidazole of 6- (the chloro- 5-FU -4- base of 2-) -4-, (2- is chloro- 5-FU -4- base) the fluoro- 1- isopropyl -2- [D of -4-₃] methyl-1 H- [5,7-D₂] benzimidazole, with 5- (4- ethyl-[D₈] piperazine Piperazine -1- ylmethyl)-pyridine -2- amine replaces 5- (4- ethyl piperazidine -1- ylmethyl)-pyridine -2- base amine, and referring to 1 step of embodiment Method preparation formula I-4 compound shown in rapid 9.

¹H-NMR (DMSO, 300MHz): δ 10.08 (s, 1H), 8.68 (d, J=3.7Hz, 1H), 8.29 (s, 1H), 8.19 ~8.22 (m, 2H), 7.64~7.70 (m, 2H), 4.80~4.89 (m, 1H), 3.43 (s, 2H), 2.64 (s, 3H), 2.30 (q, J =7.1Hz, 2H), 1.63 (d, J=6.7Hz, 6H), 0.96 (t, J=7.1Hz, 3H).

¹³C-NMR (DMSO, 300MHz): δ 155.34,154.51,153.89,152.11,150.59,148.73, 148.08,147.78,147.42,138.17,136.36,133.35,127.01,126.47,111.69,108.90,58.71, 52.12,51.47,48.05,20.88,14.49,11.98。

HRMS (M+H) m/z:515.3274.

The preparation of 5 compound I-3 of embodiment

Step 1) N- ethyl-N- nitroso piperazine

N-benzyl piperazine, and the method preparation formula XVIII-1ization referring to shown in 2 step 3 of embodiment are replaced with n-ethylpiperazine Close object.

¹H-NMR(300MHz,CDCl₃): δ 4.28 (2H, t, J=5.13Hz, piper), 3.86 (2H, t, J=5.28Hz, ), piper 2.67 (2H, t, J=5.22Hz, piper), 2.47-2.54 (2H, q, J=7.20Hz, CH₃ CH₂ ),2.43(2H,t,J =5.40Hz, piper), 1.12 (3H, t, J=7.20Hz,CH₃ CH₂)。

HRMS (ESI) m/z:148.1383.

Step 2) 4- ethyl -1- nitroso-[2,2,6,6-D₄] piperazine

N- benzyl-N- nitroso piperazine is replaced with N- ethyl-N- nitroso piperazine, and the side referring to shown in 2 step 4 of embodiment Legal system prepares standby Formula XI X-1 compound.

¹H-NMR(300MHz,CDCl₃): δ 2.64 (2H, s, piper), 2.45-2.52 (2H, q, J=7.20Hz, CH₃ CH₂ ), 2.40 (2H, s, piper), 1.11 (3H, t, J=7.20Hz,CH₃ CH₂)。

HRMS (ESI) m/z:147.1310.

Step 3) 1- ethyl-[3,3,5,5-D₄] piperazine hydrochloride

With N- ethyl-N- nitroso [2,2,6,6-D₄] piperazine replace N- benzyl-N- nitroso [2,2,6,6-D₄] piperazine, And the method referring to shown in 2 step 5 of embodiment prepares preparation formula XI-a-1 compound

¹H-NMR(300MHz,CDCl₃): δ 3.06 (4H, s, piper), 2.81-2.88 (2H, q, J=7.23Hz, CH₃ CH₂ ), 1.23 (3H, t, J=7.23Hz,CH₃ CH₂)。

HRMS (M+H) m/z:119.1479.

Step 4) 1- (the bromo- pyridin-3-yl methyl of 6-) -4- ethyl-[2,2,6,6-D₄] piperazine

Formula XI-a-1 compound (1.6g, 10.34mmol) is added in single neck bottle, sodium hydroxide (0.41g, 10.34mmol), 15mL methanol, 65 DEG C are flowed back 3 hours.It is cooled to room temperature after vulcanization acid magnesium to stir 0.5 hour, filter, filtrate is spin-dried for obtaining milky Solid.It is added in single neck bottle above-mentioned Off-white solid (1.22g, 10.32mmol), the bromo- 3- pyridine carboxaldehyde of 6- (2.30g, 12.38mmol), trimethyl orthoformate (2.26mL, 20.64mmol), formic acid (1.56mL, 41.28mmol), acetonitrile (5mL), 80 DEG C reaction overnight.2M sodium hydroxide solution tune pH is extracted to alkalinity, DCM, and anhydrous sodium sulfate is dry.Column chromatographic isolation and purification (DCM:MeOH=5:1) -5 compound of Formula XII (1.16g) is obtained, is yellow liquid.

¹H-NMR(300MHz,CDCl₃): δ 8.29 (1H, d, J=1.65Hz, Py), 7.52-7.56 (1H, dd, J= 2.40Hz, 8.16Hz, Py), 7.43 (1H, d, J=8.10Hz, Py), 3.48 (2H, s, CH₂-Py),2.40-2.51(8H,m, CH₃CH₂- piper), 1.11 (3H, t, J=7.2Hz, CH₃CH₂)。

¹³C-NMR(CDCl₃, 300MHz): δ 160.64,150.59,139.27,133.11,127.73,59.09,52.99, 52.34,51.77,11.85,11.64。

HRMS (M+H) m/z:288.1055.

Step 5) 5- (4- ethyl-[2,2,6,6-D₄] piperazine -1- ylmethyl)-pyridine -2- amine

With 1- (the bromo- pyridin-3-yl methyl of 6-) -4- ethyl-[2,2,6,6-D₄] piperazine replace 1- (the bromo- pyridin-3-yl of 6- Methyl) -4- ethyl-piperazin, and the method preparation formula V-5 compound referring to shown in 1 step 8) of embodiment.

¹H-NMR(300MHz,CDCl₃): δ 7.94 (1H, d, J=1.65Hz, Py), 7.39-7.42 (1H, dd, J= 1.50Hz, 8.31Hz, Py), 6.47 (1H, d, J=8.34Hz, Py), 4.47 (2H, s, NH₂),3.40(2H,s,CH₂-Py), 2.51-2.60(6H,m,CH₃CH₂- piper), 1.15 (3H, t, J=7.2Hz, CH₃CH₂)。

¹³C-NMR(CDCl₃, 300MHz): δ 157.73,148.50,139.20,122.76,108.39,59.43,52.20, 52.15,11.31。

HRMS (M+H) m/z:225.2004.

Step 6) [5- (4- ethyl-[2,2,6,6-D₄] piperazine -1- ylmethyl)-pyridine -2- base]-[(7- is fluoro- by 5 fluoro- 4- - 5 base of 3- isopropyl -2- methyl -3H- benzimidazole)-pyrimidine -2-base]-amine

Replacing 6- with the fluoro- 1- isopropyl -2- methyl-1 H- benzimidazole of 6- (the chloro- 5-FU -4- base of 2-) -4-, (2- is chloro- 5-FU -4- base) the fluoro- 1- isopropyl -2- [D of -4-₃] methyl-1 H- [5,7-D₂] benzimidazole, with 5- (4- ethyl-[2,2, 6,6-D₄] piperazine -1- ylmethyl)-pyridine -2- amine replaces 5- (4- ethyl piperazidine -1- ylmethyl)-pyridine -2- base amine, and reference Method preparation formula I-3 compound shown in 1 step 9) of embodiment.

¹H-NMR (DMSO, 300MHz): δ 10.12 (s, 1H), 8.68 (d, J=3.4Hz, 1H), 8.29 (s, 1H), 8.20 ~8.22 (m, 2H), 7.66~7.70 (m, 2H), 4.81~4.89 (m, 1H), 3.43 (s, 2H), 2.64 (s, 3H), 2.29~ 2.34 (m, 6H), 1.62 (d, J=6.4Hz, 6H), 0.96 (t, J=6.7Hz, 3H).

¹³C-NMR (DMSO, 300MHz): δ 155.37,154.54,153.89,152.14,150.60,148.76, 148.10,147.70,147.44,138.20,136.38,133.36,126.98,126.49,111.72,108.89,58.64, 52.12,51.55,48.06,20.89,14.51,11.90。

HRMS (M+H) m/z:511.3027.

The preparation of 6 compound I-6 of embodiment

Method one

Step 1) (6- aminopyridine -3- base) (4- ethyl piperazidine -1- base) ketone

6- amino-nicotinic acid (1.0g, 7.24mmol) is added in single neck bottle, DMF (20mL), CDI (1.41g, 8.68mmol), 10min is stirred at 70 DEG C, n-ethylpiperazine (1.84mL, 14.48mmol) is added after stirring 1h at room temperature, room temperature Lower reaction is overnight.Revolving removes solvent, and column chromatographic isolation and purification (DCM:MeOH=5:1) obtains -1 compound of Formula XIII, for white Solid (1.44g), yield 85%.

¹H-NMR(300MHz,CDCl₃): δ 8.18 (1H, s, Py), 7.53-7.57 (1H, dd, J=2.19Hz, 8.49Hz, ), Py 6.49 (1H, d, J=8.49Hz, Py), 4.77 (2H, s, NH₂),3.66(4H,s,piper),2.42-2.47(6H,m, piper-CH₂ CH₃), 1.10 (3H, t, J=7.17Hz,CH₃ CH₂)。

¹³C-NMR(CDCl₃, 300MHz): δ 168.58,159.25,147.91,137.72,121.23,107.70, 52.73,52.19,29.62,11.81。

HRMS (ESI) m/z:235.1550.

Step 2) 5- (4- ethyl piperazidine -1- base-[D₂] methyl)-pyridine -2- amine

N- benzyl-N '-acetyl group-is replaced with (6- aminopyridine -3- base) (4- ethyl piperazidine -1- base) ketone XIII-1 [3,3,5,5-D₄] piperazine, and the method preparation formula V-7 compound referring to shown in 2 step 7) of embodiment.

¹H-NMR(300MHz,CDCl₃): δ 7.95 (1H, s, Py), 7.39-7.42 (1H, dd, J=1.50Hz, 8.34Hz, ), Py 6.47 (1H, d, J=8.34Hz, Py), 4.42 (2H, s, NH₂),2.39-2.48(10H,m,piper-CH₂ CH₃),1.08 (3H, t, J=7.20Hz,CH₃ CH₂)。

¹³C-NMR(CDCl₃, 300MHz): δ 157.67,148.59,139.10,123.07,108.26,52.58,11.81.

HRMS (ESI) m/z:223.1243.

Step 3) [5- (4- ethyl piperazidine -1- base [D₂] methyl)-pyridine -2- base]-[5 fluoro- 4- (the fluoro- 3- isopropyls-of 7- - 5 base of 2- methyl -3H- benzimidazole)-pyrimidine -2-base]-amine

Replacing 6- with the fluoro- 1- isopropyl -2- methyl-1 H- benzimidazole of 6- (the chloro- 5-FU -4- base of 2-) -4-, (2- is chloro- 5-FU -4- base) the fluoro- 1- isopropyl -2- [D of -4-₃] methyl-1 H- [5,7-D₂] benzimidazole, with 5- (4- ethyl piperazidine -1- Base-[D₂] methyl)-pyridine -2- amine replaces 5- (4- ethyl piperazidine -1- ylmethyl)-pyridine -2- base amine, and referring to 1 step of embodiment It is rapid 9) shown in method preparation formula I-6 compound.

¹H-NMR (DMSO, 300MHz): δ 10.08 (s, 1H), 8.68 (d, J=3.4Hz, 1H), 8.29 (s, 1H), 8.20 ~8.23 (m, 2H), 7.66~7.70 (m, 2H), 4.81~4.89 (m, 1H), 2.64 (s, 3H), 2.35~2.50 (m, 10H), 1.63 (d, J=6.0Hz, 6H), 0.99 (t, J=6.6Hz, 3H).

¹³C-NMR (DMSO, 300MHz): δ 155.32,154.53,153.90,152.16,150.60,148.75, 148.14,147.78,147.43,138.24,136.37,133.36,126.81,126.56,111.70,108.88,52.16, 51.46,48.06,20.89,14.50,11.73。

HRMS (M+H) m/z:509.2907.

Method two

Step 1) (4- ethyl piperazidine -1- base) (6- (the fluoro- 4- of 5- (the fluoro- 1- isopropyl -2- methyl-1 H- benzimidazole-of 4- 6- yl)-pyrimidin-3-yl) ketone

By -2 compound of formula IV (428mg, 1.33mmol), -1 compound of Formula XIII (314mg, 1.33mmol), cesium carbonate (532mg, 2.66mmol) is added sequentially in dioxane (15mL), air in nitrogen displacement reaction flask, and three (two benzal are added Benzylacetone) two palladiums (30mg), the double diphenylphosphine xanthenes (50mg) of 9,9- dimethyl -4,5-.It is heated to reacting under nitrogen protection Liquid reflux.Reaction solution is cooled to room temperature after 3 hours, and 30mL ethyl acetate is added, and decompression, which filters, removes inorganic salts and metal etc. no Molten object.20mL water, liquid separation is added in filtrate, and 20mL ethyl acetate is added in water layer, and liquid separation merges organic layer, and anhydrous sodium sulfate is dry Removing solvent is concentrated under reduced pressure afterwards and obtains off-white powder.Column chromatographic purifying obtains -1 compound of Formula XIV (395mg), is white solid.

¹H-NMR (DMSO, 300MHz): δ 10.42 (s, 1H), 8.72 (d, 1H), 8.38 (s, 1H), 8.33~8.29 (m, 2H), 7.80 (d, 1H), 7.68 (d, 1H), 4.89~4.81 (m, 1H), 3.40-3.32 (m, 4H), 2.64 (s, 3H), 2.39~ 2.33(m,6H),1.63(d,6H),1.01(t,3H)。

¹³C-NMR (DMSO, 300MHz): δ 166.88,155.05,154.60,153.90,152.39,150.34, 149.02,147.60,147.00,136.96,136.42,133.35,126.30,124.78,110.89,109.0,107.26, 52.25,51.42,48.09,20.91,14.54,11.83。

HRMS (M+H) m/z:521.2474.

Step 2) [5- (4- ethyl piperazidine -1- base [D₂] methyl)-pyridine -2- base]-[5 fluoro- 4- (the fluoro- 3- isopropyls-of 7- - 5 base of 2- methyl -3H- benzimidazole)-pyrimidine -2-base]-amine

- 1 compound of Formula XIV (260mg, 0.5mmol) is dissolved in anhydrous tetrahydro furan (5mL), sequentially adds boron deuterate sodium (41mg, 1.0mmol), aluminium chloride (134mg, 1.0mmol), reaction flask add boron deuterate sodium (21mg) after being heated to reflux 2h, after It is cooled to room temperature after continuous heating reaction 1h.Be added 2M hydrochloric acid to reaction solution dissolved clarification, 70 DEG C be heated to reflux 1h after be cooled to room temperature, use 2M sodium hydroxide solution tune pH is extracted to alkalescent, ethyl acetate, and anhydrous sodium sulfate is dry.Column chromatographic isolation and purification (DCM: MeOH=5:1-6 compound of Formulas I (220mg)) is obtained, is white solid.

HRMS (M+H) m/z:509.2907.

The preparation of 7 compound I-5 of embodiment

Step 1) (6- aminopyridine -3- base) (4- Acetylpiperazine -1- base) ketone

N-ethylpiperazine, and the method preparation formula XXVI-1 referring to shown in 7 step 1) of embodiment are replaced with N- Acetylpiperazine Compound.

¹H-NMR(300MHz,CDCl₃): δ 8.18 (1H, s, Py), 7.54-7.57 (1H, dd, J=1.65Hz, 8.52Hz, ), Py 6.51 (1H, d, J=8.52Hz, Py), 4.92 (2H, s, NH₂),3.51-3.66(8H,m,piper),2.13(3H,s, CH₃)。

¹³C-NMR(CDCl₃, 300MHz): δ 169.17,159.46,147.87,137.90,120.48,107.95, 41.45,29.65,21.31。

HRMS (ESI) m/z:249.1532.

Step 2) 5- (4- [1,1-D₂] ethyl piperazidine -1- base-[D₂] methyl)-pyridine -2- amine

With (6- aminopyridine -3- base) (4- Acetylpiperazine -1- base) ketone replace N- benzyl-N '-acetyl group-[3,3, 5,5-D₄] piperazine, and with double amount [D₄] boron deuterate sodium, and the method preparation formula V-8 compound referring to shown in 2 step 7) of embodiment.

¹H-NMR(300MHz,CDCl₃): δ 7.95 (1H, d, J=1.62Hz, Py), 7.39-7.42 (1H, dd, J= 2.07Hz, 8.34Hz, Py), 6.47 (1H, d, J=8.34Hz, Py), 4.44 (2H, s, NH₂),2.40-2.50(8H,m, piper),1.07(3H,s,CH₃)。

¹³C-NMR(CDCl₃, 300MHz): δ 157.67,148.56,139.12,120.98,108.28,52.52,11.49.

HRMS (ESI) m/z:225.2022.

Step 3) [5- (4- [1,1-D₂] ethyl piperazidine -1- base-[D₂] methyl)-pyridine -2- base]-[5 fluoro- 4- (the fluoro- 3- of 7- - 5 base of isopropyl -2- methyl -3H- benzimidazole)-pyrimidine -2-base]-amine

Replacing 6- with the fluoro- 1- isopropyl -2- methyl-1 H- benzimidazole of 6- (the chloro- 5-FU -4- base of 2-) -4-, (2- is chloro- 5-FU -4- base) the fluoro- 1- isopropyl -2- [D of -4-₃] methyl-1 H- [5,7-D₂] benzimidazole, with 5- (4- [1,1-D₂] second Base piperazine -1- base-[D₂] methyl)-pyridine -2- amine replaces 5- (4- ethyl piperazidine -1- ylmethyl)-pyridine -2- base amine, and reference Method preparation formula I-5 compound shown in 1 step 9) of embodiment.

¹H-NMR (DMSO, 300MHz): δ 9.99 (s, 1H), 8.67 (d, J=2.7Hz, 1H), 8.29 (s, 1H), 8.18~ 8.21 (m, 2H), 7.66~7.70 (m, 2H), 4.81~4.89 (m, 1H), 2.64 (s, 3H), 2.35~2.50 (m, 8H), 1.63 (d, J=6.5Hz, 6H), 0.96 (s, 3H).

¹³C-NMR (DMSO, 300MHz): δ 155.34,154.50,153.88,152.12,150.58,148.74, 148.10,147.76,147.40,138.19,136.37,133.34,126.82,126.55,111.71,108.86,52.17, 48.03,20.87,14.47,11.56。

HRMS (M+H) m/z:511.3033.

The preparation of 8 compound I-7 of embodiment

[5-(4-[1,1-D₂] ethyl piperazidine -1- ylmethyl)-pyridine -2- base]-[5 fluoro- 4- (the fluoro- 3- isopropyl -2- of 7- [D₃] methyl -3H- [5,7-D₂] -5 base of benzimidazole)-pyrimidine -2-base]-amine

With 5- (4- [1,1-D₂] ethyl piperazidine -1- ylmethyl) and-pyridine -2- amine replace 5- (4- ethyl piperazidine -1- Ji Jia Base)-pyridine -2- base amine, and the method preparation formula I-7 compound referring to shown in 1 step 9) of embodiment.

¹H-NMR (DMSO, 300MHz): δ 10.09 (s, 1H), 8.69 (d, J=3.7Hz, 1H), 8.20 (d, J=6.4Hz, 2H), 7.66 (d, J=8.4Hz, 2H), 4.80~4.89 (m, 1H), 3.44 (s, 2H), 2.40~2.55 (m, 8H), 1.63 (d, J =6.8Hz, 6H), 0.96 (s, 3H).

¹³C-NMR (DMSO, 300MHz): δ 155.36,154.48,153.85,152.13,150.56,148.75, 148.09,147.78,147.43,138.19,136.28,133.40,126.95,126.32,111.71,58.74,52.36, 52.21,48.03,20.89,11.64。

HRMS (M+H) m/z:514.3224.

The preparation of 9 compound I-8 of embodiment

[5- (4- ethyl piperazidine -1- base-[D₂] methyl)-pyridine -2- base]-[5 fluoro- 4- (the fluoro- 3- isopropyl -2- [D of 7-₃] Methyl -3H- [5,7-D₂] -5 base of benzimidazole)-pyrimidine -2-base]-amine

With 5- (4- ethyl piperazidine -1- base-[D₂] methyl) and-pyridine -2- amine replace 5- (4- ethyl piperazidine -1- ylmethyl) - Pyridine -2- base amine, and the method preparation formula I-8 compound referring to shown in 1 step 9) of embodiment.

¹H-NMR (DMSO, 300MHz): δ 10.09 (s, 1H), 8.68 (d, J=3.7Hz, 1H), 8.20 (d, J=6.4Hz, 2H), 7.66 (d, J=8.4Hz, 2H), 4.80~4.89 (m, 1H), 2.40~2.55 (m, 10H), 1.63 (d, J=6.8Hz, 6H), 0.97 (t, J=6.7Hz, 3H).

HRMS (M+H) m/z:514.3212.

The preparation of 10 compound I-35 of embodiment

Step 1) 4- toluenesulfonic acid [D₅] ethyl ester

Sodium hydroxide (15.4g, 385mmol) is dissolved in water and is made into 26mL solution, D is added under condition of ice bath₆Ethyl alcohol.It will be right Toluene sulfochloride (15.8g, 84.49mmol) is dissolved in tetrahydrofuran (26mL), is slowly dropped in above-mentioned solution, is stirred at room temperature anti- It answers 2 hours.Ethyl acetate 50mL is added in reaction solution, is sufficiently stirred rear liquid separation, and water layer is added 50mL ethyl acetate again, liquid separation, Merge organic layer, anhydrous sodium sulfate drying is added, colourless viscous liquid 17.5g is concentrated under reduced pressure to obtain.

¹H-NMR (500MHz, DMSO): δ 7.80 (2H, d, J=8.0Hz, Py), 7.35 (2H, d, J=8.0Hz, Py), 2.45(3H,s,CH₃)。

¹³C-NMR (DMSO, 500MHz): 144.73,133.24,129.86,127.84,21.63.

HRMS (ESI) m/z:171.0121.

M/z 171.0121 is [M-H] of p-methyl benzenesulfonic acid^-, ethyl p-toluenesulfonate is easy to be cracked into an ion source P-methyl benzenesulfonic acid, so not acquiring the ion of toluenesulfonic acid ethyl ester.

Step 2) N- benzyl-N '-[D₅] ethyl piperazidine

Acetonitrile (10mL) is added in 25mL single-necked flask, 4- toluenesulfonic acid [D is sequentially added₅] ethyl ester (2.05g, 10mmol), benzyl diethylenediamine (1.76g, 10mmol), potassium carbonate (1.65g, 12mmol), unlatching are heated to reaction solution reflux, and 3 is small When after TLC display reaction substantially completely.Reaction solution is extracted with ethyl acetate to water layer without product, organic layer anhydrous slufuric acid acid sodium Colourless viscous liquid is rotated to obtain after drying, column chromatographic purifying obtains colourless viscous liquid 1.80g.

¹H-NMR (500MHz, DMSO): δ 7.25-7.33 (5H, m, Py), 3.54 (2H, s ,-CH₂),2.57(8H,s)。

¹³C-NMR (DMSO, 500MHz): 138.09,129.25,128.21,127.03,63.09,53.00,52.74.

HRMS (ESI) m/z:210.2116.

Step 3) N- [D₅] ethyl piperazidine

With N- benzyl-N '-[D₅] ethyl piperazidine replace N- benzyl-N '-[1,1-D₂] ethyl-[3,3,5,5-D₄] piperazine, and The method preparation formula XI-4 compound referring to shown in 2 step 8 of embodiment.

¹H-NMR (500MHz, DMSO): δ 2.93 (4H, t, J=5.0Hz), 2.43-2.46 (4H, m).

¹³C-NMR (DMSO, 500MHz): 52.77,45.25.

HRMS (ESI) m/z:120.1540.

Step 4) (6- aminopyridine -3- base) (4- [D₅] ethyl piperazidine -1- base) ketone

With N- [D₅] ethyl piperazidine replaces ethyl piperazidine, and referring to embodiment 7, the preparation of method shown in step 1 in method one - 2 compound of Formula XIII.

¹H-NMR (300MHz, DMSO): δ 7.74 (1H, d, J=3.00Hz, Py), 7.40-7.41 (1H, dd, J= 3.00Hz,Py),7.02(1H,s,Py),6.35(2H,s,NH₂), 3.49 (4H, t, J=9.00Hz, piper), 2.36 (4H, t, J=9.00Hz, piper).

¹³C-NMR (DMSO, 300MHz): 167.94,160.49,147.87,136.76,121.62,118.87,106.77, 52.24,39.78。

HRMS (ESI) m/z:240.1872.

Step 5) 5- (4- [D₅] ethyl piperazidine -1- base-[D₂] methyl)-pyridine -2- amine

With (6- aminopyridine -3- base) (4- [D₅] ethyl piperazidine -1- base) and ketone XIII-2 replace N- benzyl-N '-acetyl Base-[3,3,5,5-D₄] piperazine, and the method preparation formula V-9 compound referring to shown in 2 step 7) of embodiment.

¹H-NMR (300MHz, DMSO): δ 7.75 (1H, s, Py), 7.25-7.28 (1H, dd, J=9.00Hz, Py), 6.40 (1H, d, J=9.00Hz, Py), 5.80 (2H, s, NH₂),2.25-2.50(8H,m,piper)。

¹³C-NMR (DMSO, 300MHz): δ 158.14,148.92,137.56,122.04,109.64,55.96,11.59.

HRMS (ESI) m/z:227.2127.

Step 6) [5- (4- [D₅] ethyl piperazidine -1- base-[D₂] methyl)-pyridine -2- base]-[5 fluoro- 4- (the fluoro- 3- isopropyls of 7- - 5 base of base -2- methyl -3H- benzimidazole)-pyrimidine -2-base]-amine

Replacing 6- with the fluoro- 1- isopropyl -2- methyl-1 H- benzimidazole of 6- (the chloro- 5-FU -4- base of 2-) -4-, (2- is chloro- 5-FU -4- base) the fluoro- 1- isopropyl -2- [D of -4-₃] methyl-1 H- [5,7-D₂] benzimidazole, with 5- (4- [D₅] ethyl piperazine Piperazine -1- base-[D₂] methyl)-pyridine -2- amine replaces 5- (4- ethyl piperazidine -1- ylmethyl)-pyridine -2- base amine, and referring to implementation Method preparation formula I-35 compound shown in 1 step 9) of example.

¹H-NMR (DMSO, 300MHz): δ 10.10 (s, 1H), 8.69 (s, 1H), 8.30 (s, 1H), 8.19~8.22 (m, 2H), 7.66~7.70 (m, 2H), 4.81~4.89 (m, 1H), 2.64 (s, 3H), 2.35~2.50 (m, 8H), 1.63 (d, J= 5.2Hz,6H)。

HRMS (M+H) m/z:514.3229.

The preparation of 11 compound I-36 of embodiment

Step 1) [5- (4- [D₅] ethyl piperazidine -1- base-[D₂] methyl)-pyridine -2- base]-[5 fluoro- 4- (the fluoro- 3- isopropyls of 7- Base -2- [D₃] methyl -3H- [5,7-D₂] -5 base of benzimidazole)-pyrimidine -2-base]-amine

With 5- (4- [D₅] ethyl piperazidine -1- base-[D₂] methyl) and-pyridine -2- amine replace 5- (4- ethyl piperazidine -1- Ji Jia Base)-pyridine -2- base amine, and the method preparation formula I-36 compound referring to shown in 1 step 9) of embodiment.

¹H-NMR (DMSO, 300MHz): δ 10.12 (s, 1H), 8.69 (d, J=3.8Hz, 1H), 8.20 (d, J=6.5Hz, 2H), 7.66 (d, J=8.9Hz, 1H), 4.80~4.89 (m, 1H), 2.40~2.55 (m, 8H), 1.63 (d, J=6.8Hz, 6H)。

¹³C-NMR (DMSO, 300MHz): δ 155.39,154.54,153.87,152.20,150.57,148.78, 148.18,147.84,147.49,138.28,136.29,126.80,126.34,111.75,52.20,48.08,20.93。

HRMS (M+H) m/z:519.3531.

The preparation of 12 compound I-37 of embodiment

Step 1) [5- (4- [1,1-D₂] ethyl piperazidine -1- base-[D₂] methyl)-pyridine -2- base]-[5 fluoro- 4- (the fluoro- 3- of 7- Isopropyl -2- [D₃] methyl -3H- [5,7-D₂] -5 base of benzimidazole)-pyrimidine -2-base]-amine

With 5- (4- [1,1-D₂] ethyl piperazidine -1- base-[D₂] methyl) and-pyridine -2- amine replace 5- (4- ethyl piperazidine -1- base Methyl)-pyridine -2- base amine, and the method preparation formula I-37 compound referring to shown in 1 step 9) of embodiment.

¹H-NMR (DMSO, 300MHz): δ 10.10 (s, 1H), 8.69 (d, J=3.9Hz, 1H), 8.20 (d, J=5.2Hz, 2H),7.66(dd,J₁=2.1Hz, J₂=8.7Hz, 1H), 4.80~4.89 (m, 1H), 2.30~2.42 (m, 8H), 1.63 (d, J =6.9Hz, 6H), 0.95 (s, 3H).

¹³C-NMR (DMSO, 300MHz): δ 155.37,154.54,153.86,152.17,150.57,148.97, 148.78,147.85,147.50,138.26,126.90,111.77,52.40,48.07,20.92,11.77。

HRMS (M+H) m/z:516.3344.

The preparation of 13 compound I-38 of embodiment

Step 1) (6- aminopyridine -3- base) (4- acetyl group-[D₈] piperazine -1- base) ketone

With N- acetyl group-[D₈] piperazine replaces n-ethylpiperazine, and the method preparation formula referring to shown in 8 step 1) of embodiment XXVI-2 compound.

¹H-NMR (300MHz, DMSO): δ 8.19 (1H, d, J=2.0Hz, Py), 7.56-7.58 (1H, dd, J₁=2.0Hz, J₂=8.5Hz, Py), 6.52 (1H, d, J=8.5Hz, Py), 4.90 (2H, s, NH₂),2.14(3H,s,CH₃)。

¹³C-NMR (DMSO, 300MHz): δ 169.28,159.58,148.07,137.88,120.46,107.91,21.40.

HRMS (ESI) m/z:257.1858.

Step 2) 5- (4- [1,1-D₂] ethyl-[D₈] piperazine -1- base-[D₂] methyl)-pyridine -2- amine

With (6- aminopyridine -3- base) (4- acetyl group-[D₈] piperazine -1- base) and ketone replace N- benzyl-N '-acetyl group - [3,3,5,5-D₄] piperazine, and with double amount [D₄] boron deuterate sodium, and the method preparation formula V-10 referring to shown in 2 step 7) of embodiment Compound.

¹H-NMR (300MHz, DMSO): δ 7.74 (1H, s, Py), 7.24-7.27 (1H, dd, J=9.00Hz, Py), 6.38 (1H, d, J=9.00Hz, Py), 5.78 (2H, s, NH₂),0.94(3H,s,CH₃)。

¹³C-NMR (DMSO, 300MHz): δ 158.52,148.36,137.48,122.10,109.14,61.82,10.65.

HRMS (ESI) m/z:232.2441.

Step 3) [5- (4- [1,1-D₂] ethyl-[D₈] piperazine -1- base-[D₂] methyl)-pyridine -2- base]-[5 fluoro- 4- (7- Fluoro- 3- isopropyl -2- [D₃] methyl -3H- [5,7-D₂] -5 base of benzimidazole)-pyrimidine -2-base]-amine

With 5- (4- [1,1-D₂] ethyl [D₈] piperazine -1- base-[D₂] methyl) and-pyridine -2- amine replace 5- (4- ethyl piperazidine - 1- ylmethyl)-pyridine -2- base amine, and the method preparation formula I-38 compound referring to shown in 1 step 9) of embodiment.

¹H-NMR (DMSO, 300MHz): δ 10.10 (s, 1H), 8.69 (d, J=3.9Hz, 1H), 8.20 (d, J=5.0Hz, 2H),7.66(dd,J₁=2.1Hz, J₂=8.7Hz, 1H), 4.80~4.89 (m, 1H), 1.63 (d, J=6.9Hz, 6H), 0.96 (s,3H)。

¹³C-NMR (DMSO, 300MHz): δ 155.42,154.56,153.87,152.19,148.79,148.18, 147.86,147.51,138.29,126.86,111.78,48.08,20.93,11.61。

HRMS (M+H) m/z:524.3854.

The preparation of 14 compound I-24 of embodiment

Step 1) N- [D₇] isopropyl-acetamide

By D₈Isopropanol (5.7g, 83.2mmol) is dissolved in acetonitrile (85.38g, 2.1mol), is heated to 60 DEG C, disposably The 10ml concentrated sulfuric acid is added, reaction system very exothermic closes heating, reflux unit reflux, water flow increases；50 are down to temperature DEG C, heating is opened, aforesaid operations are repeated, continues enriching sulfuric acid, concentrated sulfuric acid 30ml (0.56mol) is added altogether.TLC tracking, to anti- It has been answered that, reaction system has been poured into ice water, with NaOH aqueous solution neutralization reaction system.It is extracted twice with DCM, with NaCl aqueous solution It washes twice, dry, vacuum distillation removes solvent, ether dilution, and 10g K is added₂CO₃, room temperature mashing 2h.Filtering is depressurized dense Contracting obtains 8.7g light yellow oil.Yield: 96%.

¹H-NMR(CDCl₃, 300MHz): δ 1.95 (s, 3H).

¹³C-NMR(CDCl₃, 300MHz): δ 171.81,77.0,24.72,23.17.

HRMS (M+H) m/z:109.1349.

Step 2) N- (the bromo- 2,6- difluorophenyl of 4-)-N '-[D₇] isopropyl-second narrows

With N- [D₇] isopropyl-acetamide replaces N- isopropyl-acetamide, and the method referring to shown in 1 step 2) of embodiment Preparation formula IX-2 compound.

¹H-NMR(CDCl₃, 300MHz): δ 7.03 (d, J=6.12Hz, 2H), 4.45 (s, 1H), 1.76 (s, 3H).

¹³C-NMR(CDCl₃, 300MHz): δ 157.19,153.98,115.32,115.17,115.09,114.95, 77.42,77.00,76.57,18.72,18.66。

HRMS (M+H) m/z:298.0751.

Fluoro- 1- [the D of the bromo- 4- of step 3) 6-₇] isopropyl -2- methyl-1 H- benzimidazole

With N- (the bromo- 2,6- difluorophenyl of 4-)-N '-[D₇] isopropyl-second narrows instead of N- (the bromo- 2,6- difluorophenyl of 4-)- N '-isopropyl-second is narrowed, and the method preparation formula II-3 compound referring to shown in 1 step 3) of embodiment.

¹H-NMR(CDCl₃, 300MHz): δ 7.41 (d, J=1.17Hz, 1H), 7.08 (dd, J₁=1.26Hz, J₂= 9.6Hz,1H),2.63(s,3H)。

¹³C-NMR(CDCl₃, 300MHz): δ 154.76,151.94,111.45,111.17,110.34,110.28, 77.42,77.00,76.57,14.78。

HRMS (M+H) m/z:278.0684.

Fluoro- 1- [the D of step 4) 4-₇] isopropyl -2- methyl -6- (4,4,5,5- tetramethyl-[1,3,2] dioxa boron heterocycle Pentane -2- base) -1H- benzimidazole

With the fluoro- 1- [D of the bromo- 4- of 6-₇] isopropyl -2- methyl-1 H- benzimidazole replace the fluoro- 1- isopropyl -2- of the bromo- 4- of 6- [D₃] methyl-1 H- [5,7-D₂] benzimidazole, and the method preparation formula III-3 compound referring to shown in 1 step 5) of embodiment.

¹H-NMR(CDCl₃, 300MHz): δ 7.69 (s, 1H), 7.37 (m, 1H), 2.68 (s, 3H), 1.36 (s, 12H).

¹³C-NMR (DMSO, 300MHz): δ 152.03,113.60,113.56,113.18,112.97,84.03,77.41, 76.99,76.57,24.87,14.84。

HRMS (M+H) m/z:326.2423.

Step 5) 6- (the chloro- 5-FU -4- base of 2-) fluoro- 1- [D of -4-₇] isopropyl -2- methyl-1 H- benzimidazole

With the fluoro- 1- [D of 4-₇] isopropyl -2- methyl -6- (4,4,5,5- tetramethyl-[1,3,2] dioxaborolan alkane - 2- yl) -1H- benzimidazole replace the fluoro- 1- isopropyl -2- [D of 4-₃] methyl -6- (4,4,5,5- tetramethyl-[1,3,2] dioxa Boron heterocycle pentane -2- base) -1H- [5,7-D₂] benzimidazole, and the method preparation formula IV-3ization referring to shown in 1 step 6) of embodiment Close object.

¹H-NMR(CDCl₃, 300MHz): δ 8.51 (d, J=3.42Hz, 1H), 8.15 (s, 1H), 7.80 (d, J= 11.49Hz),2.70(s,3H)。

¹³C-NMR(CDCl₃, 300MHz): δ 156.63,154.22,151.66,148.57,148.21,108.88, 108.76,108.51,108.40,108.24,108.13,15.17。

HRMS (M+H) m/z:330.1079.

Step 6) [5- (4- ethyl-piperazin -1- ylmethyl)-pyridine -2- base]-[the 5 fluoro- 4- (fluoro- 3- [D of 7-₇] isopropyl- - 5 base of 2- methyl -3H- benzimidazole)-pyrimidine -2-base]-amine

With 6- (the chloro- 5-FU -4- base of the 2-) fluoro- 1- [D of -4-₇] isopropyl -2- methyl-1 H- benzimidazole replace 6- (the chloro- 5-FU -4- base of 2-) fluoro- 1- isopropyl -2- [D of -4-₃] methyl-1 H- [5,7-D₂] benzimidazole, and referring to embodiment Method preparation formula I-24 compound shown in 1 step 9).

¹H-NMR(CDCl₃, 300MHz): δ 8.41 (m, 2H), 8.26 (s, 1H), 8.18 (s, 1H), 7.79 (d, J= 11.64Hz, 1H), 7.68 (m, 1H), 3.53 (s, 2H), 2.69~2.64 (m, 5H), 2.56 (t, J=7.29Hz, 8H), 1.18 (t, J=7.14Hz, 3H).

¹³C-NMR(CDCl₃, 300MHz): δ 154.69,153.11,151.64,148.84,146.81,146.45, 138.48,135.90,127.65,126.89,126.41,108.23,108.12,108.08,107.79,107.52,107.42, 76.93,59.07,51.91,51.73,51.53,14.51,10.78。

HRMS (M+H) m/z:514.3225.

The preparation of 15 compound I-39 of embodiment

Step 1) [5- (4- [D₅] ethyl piperazidine -1- ylmethyl)-pyridine -2- base]-[the 5 fluoro- 4- (fluoro- 3- [D of 7-₇] isopropyl - 5 base of base -2- methyl -3H- benzimidazole)-pyrimidine -2-base]-amine

With 6- (the chloro- 5-FU -4- base of the 2-) fluoro- 1- [D of -4-₇] isopropyl -2- methyl-1 H- benzimidazole replace 6- (the chloro- 5-FU -4- base of 2-) fluoro- 1- isopropyl -2- [D of -4-₃] methyl-1 H- [5,7-D₂] benzimidazole, with 5- (4- [D₅] Ethyl piperazidine -1- base-methyl)-pyridine -2- amine replaces 5- (4- ethyl piperazidine -1- ylmethyl)-pyridine -2- base amine, and referring to reality Apply method preparation formula I-39 compound shown in 1 step 9) of example.

¹H-NMR (DMSO, 500MHz): δ 10.08 (s, 1H), 8.68 (d, J=3.5Hz, 1H), 8.29 (s, 1H), 8.20 ~8.21 (m, 2H), 7.65~7.69 (m, 2H), 3.44 (s, 2H), 2.64 (s, 3H), 2.34~2.49 (m, 8H).

¹³C-NMR(DMSO,500MHz):155.86,153.74,152.66,151.97,150.55,149.55,148.63, 138.74,136.82,133.69,127.47,112.26,109.33,107.49,59.25,52.85,15.01。

HRMS (M+H) m/z:519.3573.

The preparation of 16 compound I-17 of embodiment

Step 1) N- benzyl-N '-[D₅] ethyl [3,3,5,5-D₄] piperazine

With N- benzyl-[3,3,5,5-D₄] piperazine replaces benzyl diethylenediamine, and prepared by the method referring to shown in 11 step 2 of embodiment Formula X XIV-4 compound.

Step 2) N- [1,1,2,2,2-D₅] ethyl-[2,2,6,6-D₄] piperazine

With N- benzyl-N '-[D₅] ethyl-[3,3,5,5-D₄] piperazine replace N- benzyl-N '-[1,1-D₂] ethyl-[3,3, 5,5-D₄] piperazine, and the method preparation formula XI-5 compound referring to shown in 2 step 8 of embodiment.

¹H-NMR(CDCl₃, 500MHz): δ 2.86 (s, 4H).

¹³C-NMR(CDCl₃, 500MHz): δ 45.74.

HRMS (M+H) m/z:124.1788.

Step 3) 1- (the bromo- pyridin-3-yl methyl of 6-) -4- [1,1,2,2,2-D₅] ethyl-[3,3,5,5-D₄] piperazine

With N- [D₅] ethyl-[2,2,6,6-D₄] piperazine replace N- [1,1-D₂] ethyl-[2,2,6,6-D₄] piperazine, and join According to method preparation formula XII-7 compound shown in 2 step 9) of embodiment.

¹H-NMR(CDCl₃, 500MHz): δ 8.33 (d, J=2Hz, 1H), 7.47 (d, J=8.2Hz, 1H), 7.50 (dd, J =8.2Hz, 2.0Hz, 1H), 3.6 (s, 2H), 3.03 (s, 2H), 2.90 (s, 2H).

¹³C-NMR(CDCl₃, 500MHz): δ 150.7,141.5,139.2,131.9,128.0,58.4,50.6,49.0, 49.0,37.0。

HRMS (M+H) m/z:293.1619.

Step 4) 5- (4- [1,1,2,2,2-D₅] ethyl-[3,3,5,5--D₄] piperazine -1- ylmethyl)-pyridine -2- base amine

With 1- (the bromo- pyridin-3-yl methyl of 6-) -4- [D₅] ethyl-[3,3,5,5-D₄] piperazine replace 1- (the bromo- pyridine-of 6- 3- ylmethyl) -4- [1,1-D₂] ethyl-[3,3,5,5-D₄] piperazine, and the method preparation formula V- referring to shown in 2 step 10 of embodiment 12 compounds.

¹H-NMR(CDCl₃, 500MHz): δ 7.97 (d, J=2.2Hz, 1H), 7.43 (dd, J=8.4Hz, 2.2Hz, 1H), 6.49 (d, J=8.4Hz, 1H), 4.43 (s, 2H), 3.40 (s, 2H), 2.50 (s, 4H).

¹³C-NMR(CDCl₃, 500MHz): δ 157.7,148.6,139.2,123.2,108.4,59.8,52.4.

HRMS (ESI) m/z:230.2323.

Step 5) [5- (4- [1,1,2,2,2-D₅] ethyl-[3,3,5,5--D₄] piperazine -1- base-methyl)-pyridine -2- Base]-[5 fluoro- 4- (fluoro- -5 base of 3- isopropyl -2- methyl -3H- benzimidazole of 7-)-pyrimidine -2-base]-amine

Replacing 6- with the fluoro- 1- isopropyl -2- methyl-1 H- benzimidazole of 6- (the chloro- 5-FU -4- base of 2-) -4-, (2- is chloro- 5-FU -4- base) the fluoro- 1- isopropyl -2- [D of -4-₃] methyl-1 H- [5,7-D₂] benzimidazole, with 5- (4- [D₅] ethyl [2, 2,6,6-D₄] piperazine -1- base-methyl) and-pyridine -2- amine replace 5- (4- ethyl piperazidine -1- ylmethyl)-pyridine -2- base amine, and The method preparation formula I-17 compound referring to shown in 1 step 9) of embodiment.

¹H-NMR(CDCl₃, 500MHz): δ 8.48 (m, 1H), 8.41 (d, J=8.25Hz, 1H), 8.29 (d, J=1.5Hz, 1H), 8.21 (d, J=1.0Hz, 1H), 7.81 (d, J=11.5Hz, 1H), 7.71 (m, 1H), 4.76 (quintet, J=7Hz, 1H), 3.53 (s, 2H), 2.71 (s, 3H), 2.65 (s, 4H), 1.73 (d, J=7Hz, 6H).

¹³C-NMR(CDCl₃, 500MHz): 154.32,153.62,152.08,150.00,148.72,136.47, 136.39,134.18,134.05,127.42,111.42,108.75,108.24,108.18,108.02,59.77,52.40, 48.66,21.48,15.07。

HRMS (M+H) m/z:516.3403.

The preparation of 1 compound I-2 of reference implementation example

Step 1) 1- (the bromo- pyridin-3-yl methyl of 6-) -4- acetyl group-piperazine

N-ethylpiperazine, and the method preparation formula XXV-2ization referring to shown in 1 step 7) of embodiment are replaced with N- Acetylpiperazine Close object.

¹H-NMR(300MHz,CDCl₃): δ 8.29 (1H, d, J=1.65Hz, Py), 7.55-7.58 (1H, dd, J= 1.74Hz, 8.10Hz, Py), 7.46 (1H, d, J=8.10Hz, Py), 3.62 (2H, t, J=4.77Hz, piper), 3.49 (2H, s,CH₂- Py), 3.46 (2H, t, J=4.86Hz, piper), 2.40-2.44 (4H, m, piper), 2.08 (3H, s, CH₃)。

¹³C-NMR(CDCl₃, 300MHz): δ 150.48,141.03,139.20,132.61,127.88,59.10,52.93, 52.65,46.10,41.24,21.24。

HRMS (M+H) m/z:298.0549.

Step 2) 1- (the bromo- pyridin-3-yl methyl of 6-) -4- [1,1-D₂] ethyl piperazidine

N- benzyl-N '-acetyl group-[3,3,5,5- is replaced with 1- (the bromo- pyridin-3-yl methyl of 6-) -4- acetyl group-piperazine D₄] piperazine, and the method preparation formula XII-6 compound referring to shown in 2 step 7) of embodiment.

¹H-NMR(300MHz,CDCl₃): δ 8.29 (1H, d, J=1.74Hz, Py), 7.52-7.56 (1H, dd, J= 1.98Hz, 8.10Hz, Py), 7.43 (1H, d, J=8.10Hz, Py), 3.47 (2H, s, CH₂),2.50(8H,s,piper), 1.08(3H,s,CH₃ CD₂)。

¹³C-NMR(CDCl₃, 300MHz): δ 150.59,140.73,139.28,133.18,127.73,59.25,52.84, 52.54,11.56。

HRMS (M+H) m/z:286.0879.

Step 3) 5- (4- [1,1-D₂] ethyl piperazidine -1- ylmethyl)-pyridine -2- amine

With 1- (the bromo- pyridin-3-yl methyl of 6-) -4- [1,1-D₂] ethyl piperazidine replace 1- (the bromo- pyridin-3-yl first of 6- Base) -4- ethyl-piperazin, and the method preparation formula V-6 compound referring to shown in 1 step 8) of embodiment.

¹H-NMR(300MHz,CDCl₃): δ 7.95 (1H, d, J=1.65Hz, Py), 7.39-7.42 (1H, dd, J= 1.50Hz, 8.31Hz, Py), 6.47 (1H, d, J=8.31Hz, Py), 4.42 (2H, s, NH₂),3.37(2H,s,CH₂),2.47 (8H,s,piper),1.05(3H,s,CH₃ CD₂)。

¹³C-NMR(CDCl₃, 300MHz): δ 157.63,148.58,139.11,123.27,108.26,59.79,52.77, 52.63,11.65。

HRMS (ESI) m/z:223.1890.

Step 4) [5- (4- [1,1-D₂] ethyl piperazidine -1- ylmethyl)-pyridine -2- base]-[5 fluoro- 4- (the fluoro- 3- isopropyls of 7- - 5 base of base -2- methyl -3H- benzimidazole)-pyrimidine -2-base]-amine

6- (2- is replaced with the fluoro- 1- isopropyl -2- methyl-1 H- benzimidazole of 6-- (the chloro- 5-FU -4- base of 2-) -4- Chloro- 5-FU -4- base) the fluoro- 1- isopropyl -2- [D of -4-₃] methyl-1 H- [5,7-D₂] benzimidazole, with 5- (4- [1,1-D₂] Ethyl piperazidine -1- ylmethyl)-pyridine -2- amine replaces 5- (4- ethyl piperazidine -1- ylmethyl)-pyridine -2- base amine, and referring to reality Apply method preparation formula I-2 compound shown in 1 step 9) of example.

¹H-NMR (DMSO, 300MHz): δ 10.06 (s, 1H), 8.69 (d, J=3.4Hz, 1H), 8.29 (s, 1H), 8.20 ~8.22 (m, 2H), 7.65~7.69 (m, 2H), 4.81~4.89 (m, 1H), 3.47 (s, 2H), 2.65 (s, 3H), 2.40~ 2.55 (m, 8H), 1.63 (d, J=6.4Hz, 6H), 0.84 (s, 3H).

HRMS (M+H) m/z:509.2899.

The preparation of 2 compound I-16 of reference implementation example

Step 1) 5- (4- [D₅] ethyl piperazidine -1- ylmethyl)-pyridine -2- amine

With (6- aminopyridine -3- base) (4- [D₅] ethyl piperazidine -1- base) and ketone XIII-2 replace N- benzyl-N '-acetyl Base-[3,3,5,5-D₄] piperazine, and the method preparation formula V-11 compound referring to shown in 2 step 7) of embodiment.

¹H-NMR (300MHz, DMSO): δ 7.74 (1H, s, Py), 7.24-7.27 (1H, dd, J=9.00Hz, Py), 6.39 (1H, d, J=9.00Hz, Py), 5.76 (2H, s, NH₂),3.23(2H,s,-CH₂),2.32-2.50(8H,m,piper)。

¹³C-NMR(DMSO,300MHz):158.95,147.97,138.07,120.75,107.52,59.09,39.52。

HRMS (ESI) m/z:226.2084.

Step 2) [5- (4- [D₅] ethyl piperazidine -1- ylmethyl)-pyridine -2- base]-[5 fluoro- 4- (the fluoro- 3- isopropyls-of 7- - 5 base of 2- methyl -3H- benzimidazole)-pyrimidine -2-base]-amine

Replacing 6- with the fluoro- 1- isopropyl -2- methyl-1 H- benzimidazole of 6- (the chloro- 5-FU -4- base of 2-) -4-, (2- is chloro- 5-FU -4- base) the fluoro- 1- isopropyl -2- [D of -4-₃] methyl-1 H- [5,7-D₂] benzimidazole, with 5- (4- [D₅] ethyl piperazine Piperazine -1- base-methyl)-pyridine -2- amine replaces 5- (4- ethyl piperazidine -1- ylmethyl)-pyridine -2- base amine, and referring to embodiment 1 The I-16 compound of method preparation formula shown in step 9).

¹H-NMR (DMSO, 500MHz): δ 10.09 (s, 1H), 8.68 (d, J=4.0Hz, 1H), 8.29 (s, 1H), 8.20 ~8.21 (m, 2H), 7.66~7.69 (m, 2H), 4.82~4.87 (m, 1H), 3.45 (s, 2H), 2.64 (s, 3H), 2.35~ 2.48 (m, 8H), 1.63 (d, J=7.0Hz, 6H).

¹³C-NMR(DMSO,500MHz):δ155.88,153.75,152.71,151.77,150.62,149.96, 148.26,138.78,136.79,133.82,127.30,112.27,107.59,59.13,52.52,48.59,21.42, 15.03。

HRMS (M+H) m/z:512.3152.

The analysis of embodiment 17CDK4 inhibitory activity

Experimental procedure

Prepare ATP and peptide substrate Ulight-4E-BP1 (Thr37/46) (PerkinElmer company Cat NO.TRF0128-D) solution obtains being diluted in 50mM HEPES, 10mM MgCl₂, 10mM EGTA, 2mM DTT, 0.01% Final concentration of 70 μM of ATP and 50nM Ulight-4E-BP1 (Thr37/46) in the kinase buffer liquid of Tween 20.Prepare enzyme Solution obtains CDK4 enzyme (the CARNA Biosciences company for being diluted in final concentration of 0.5ng/ μ L in above-mentioned kinase buffer liquid Cat NO.04-105).By test compound of the 2 μ L in 0.02%DMSO, 4 μ L ATP and peptide substrate Ulight-4E-BP1 (Thr37/46) solution and 4 μ L enzyme solutions mix in 384 orifice plate of low volume.Test compound is serially diluted with 1:3 In 0.02%DMSO, the curve of 8 points is generated, initial concentration 400nM does not add the independent 0.02%DMSO of test compound Buffer control, kinase buffer liquid as be not present enzyme activity when background signal, reagent is mixed, centrifugation 1 minute (2500 turns/ Minute) and sealer after 25 DEG C incubate 60 minutes, by addition 5 μ L 40mM EDTA terminate reaction (final concentration 10mM), after 5 μ L 8nM (final concentration 2nM) Eu-anti-P-4E-BP1 (Thr37/46) (PerkinElmer company Cat are added NO.TRF0216-D it) into detection plate, mixes, continues to incubate 60 points at 25 DEG C after being centrifuged 1 minute (2500 revs/min) sealer Clock, after be put into multi-functional plate reader (Molecular Devices company model Paradigm) using HTRF module detect numerical value.

Experimental result

1. compound of table inhibits IC to CDK4₅₀It is worth (nM)

Using four parameter fitting modes, IC is calculated to the inhibiting rate matched curve of testing result₅₀Value, sees the above table 1, is screened I-1, I-2, I-3, I-4, I-5, I-6, I-7, I-8, I-9 and I-10 and positive compound LY-2835219 result it is close, Less than 2nM.Experimental result confirms that the above compound is excellent CDK4 inhibitor.

The analysis of 18 CDK6 inhibitory activity of embodiment

Experimental procedure

Prepare ATP and peptide substrate Ulight-4E-BP1 (Thr37/46) (PerkinElmer company Cat NO.TRF0128-D) solution obtains being diluted in 50mMHEPES, 10mM MgCl₂, 10mM EGTA, 2mM DTT, 0.01% Final concentration of 70 μM of ATP and 50nMUlight-4E-BP1 (Thr37/46) in the kinase buffer liquid of Tween 20.It is molten to prepare enzyme Liquid obtains CDK6 enzyme (the CARNA Biosciences company for being diluted in final concentration of 0.5ng/ μ L in above-mentioned kinase buffer liquid Cat NO.04-107).By test compound of the 2 μ L in 0.02%DMSO, 4 μ L ATP and peptide substrate Ulight-4E-BP1 (Thr37/46) solution and 4 μ L enzyme solutions mix in 384 orifice plate of low volume.Test compound is serially diluted with 1:3 In 0.02%DMSO, the curve of 7 points is generated, initial concentration 100nM does not add the independent 0.02%DMSO of test compound Buffer control, kinase buffer liquid as be not present enzyme activity when background signal, reagent is mixed, centrifugation 1 minute (2500 turns/ Minute) and sealer after 25 DEG C incubate 60 minutes, by addition 5 μ L 40mM EDTA terminate reaction (final concentration 10mM), after 5 μ L 8nM (final concentration 2nM) Eu-anti-P-4E-BP1 (Thr37/46) (PerkinElmer company Cat are added NO.TRF0216-D it) into detection plate, mixes, continues to incubate 60 points at 25 DEG C after being centrifuged 1 minute (2500 revs/min) sealer Clock, after be put into multi-functional plate reader (Molecular Devices company model Paradigm) using HTRF module detect numerical value.

Experimental result

2. compound of table inhibits IC to CDK6₅₀It is worth (nM)

Using four parameter fitting modes, IC is calculated to the inhibiting rate matched curve of testing result₅₀Value, sees the above table 2, is screened I-1, I-2, I-3, I-4, I-5, I-6, I-7, I-8, I-9, I-10 and reference material LY-2835219 compound inhibit CDK6 living Similar temperament, respectively less than 5nM.As a result confirm that the above compound is excellent CDK6 inhibitor.

The stability study in people's hepatomicrosome of embodiment 19

300 μ L final temperature is incubated in system, and 30 μ L hepatomicrosomes (protein concentration: 0.15mg/mL), 30 μ L NADPH+ are contained MgCl₂, 3 μ L substrates, 237 μ L PBS buffer solution.Wherein the ratio of organic solvent (acetonitrile) is 1%.Each kind does 2 parts, often Part 0.3mL.Every pipe first prepares the substrate that total volume is 270 μ L and the mixing liquid and NADPH of enzyme incubate 5min in 37 DEG C of pre-temperatures respectively Afterwards, 30 μ L NADPH+MgCl are added₂Mixing takes out the ice acetonitrile of 50 μ L containing the internal standards respectively at 0,15,30,60,120min 300 μ L terminate reaction.In addition 3 parts of each 300 μ L of blank (KB) are respectively, and NADPH respectively KB1: is not added；KB2: substrate is not added； KB3: not enzyme.

It draws 50 μ L temperature and incubates sample, the ice acetonitrile precipitation of 300 μ L containing the internal standards is added, be vortexed after concussion 5min, centrifugation (8800rpm, 4 DEG C) 10min.100 μ L of Aspirate supernatant is into sample injection bottle, and sample introduction is analyzed by 0.5 μ L.Related compound is micro- in people liver Parameter is eliminated in plastochondria is shown in Table 3.

The external people's hepatomicrosome metabolic stability of 3 related compound of table

Pharmacokinetic Evaluation in 20 rat of embodiment

SD rat, 200~220g of weight are randomly divided into 7 groups, every group 3, give 10mg/kg respectively after adapting to 3-5 days The compound of dosage: Abemaciclib, I-4, I-5, I-8, I-9, I-10.

Fasting 12h before animal subject (SD rat) administration, 4h is tested in front and back and experimentation certainly to food after administration By drinking water.

After gastric infusion, in 0min, 15min, 30min, 1h, 3h, 4h, 5h, 6h, 8h, 10h, for 24 hours in eye socket blood is taken 0.10mL or so, EDTA-K2 are anticoagulant, immediately place in ice blood sample, 4 DEG C, 4000rpm are transferred in 30min, 10min condition Lower centrifugal separation plasma.It is to be measured in -20 DEG C of preservations immediately after the whole blood plasma of collection.

It draws 40 μ L test plasma samples and marks bent sample, the methanol solution of 200 μ L containing the internal standards is added, oscillation mixes 5min, 13000rpm are centrifuged 10min, take 80 μ L of supernatant, draw 1 μ L and measure for LC/MS/MS, record chromatogram.

Pass through the oral administration biaavailability of Rats pharmacokinetics experimental evaluation the compounds of this invention.Some representativenesses The medicine of compound is as shown in the table for parameter.

The medicine of 4 related compound of table is for parameter

Claims

1. compound as follows or its pharmaceutically acceptable salt:

2. compound as follows or its pharmaceutically acceptable salt:

3. a kind of pharmaceutical composition, it includes the compound of any one of claim 1-2 or its pharmaceutically acceptable salt, with And one or more pharmaceutically acceptable carriers or excipient.

4. the compound of any one of claim 1-2 or the pharmaceutical composition of claim 3 are in preparation treating cancer drug Purposes.

5. the purposes of claim 4, wherein the cancer is selected from colorectal cancer, breast cancer, lung cancer, prostate cancer, colloid Blastoma, lymphoma mantle cell, chronic myelocytic leukemia or acute myeloblastic leukemia.