CN105153122B - [(indol-3-yl) pyrimidine -2-base] aminophenyl propyl- 2- alkenylamide derivatives and salt, preparation method, application - Google Patents
[(indol-3-yl) pyrimidine -2-base] aminophenyl propyl- 2- alkenylamide derivatives and salt, preparation method, application Download PDFInfo
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- CN105153122B CN105153122B CN201510534307.4A CN201510534307A CN105153122B CN 105153122 B CN105153122 B CN 105153122B CN 201510534307 A CN201510534307 A CN 201510534307A CN 105153122 B CN105153122 B CN 105153122B
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- C07D403/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00
- C07D403/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings
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Abstract
The present invention provides [2 base of (3 base of indoles) pyrimidine] aminophenyl the third 2 alkenylamide derivative and its salt, preparation method, application, the third 2 alkenylamide derivative of [2 base of (3 base of indoles) pyrimidine] aminophenyl has Formulas I structure.Deuterium carbon key in the third 2 alkenylamide derivative of [2 base of (3 base of indoles) pyrimidine] aminophenyl provided by the invention makes it, and decomposition rate is slow in vivo, extend the half-life period of drug, increase the blood concentration of drug, its dosage is finally reduced, its toxic side effect is reduced.The experimental results showed that：The 9291 D9 mesylates of deuterated derivative AZD compare 9291 mesylates of AZD, CmaxIt is 1.32 times of 9291 mesylates of AZD；Its exposed amount is 1.41 times of 9291 mesylates of AZD；Its elimination half-life period is 1.31 times of 9291 mesylates of AZD.
Description
Technical field
The present invention relates to field of pharmaceutical chemistry technology, more particularly to [(indol-3-yl) pyrimidine -2-base] aminophenyl propyl-
2- alkenylamide derivatives and its salt, preparation method, application.
Background technology
In worldwide, the malignant tumour that incidence and the death rate rank the first is lung cancer, wherein most common right and wrong
Small Cell Lung Cancer.EGF-R ELISA -1 (epidermal growth factor receptor, EGFR) is often expressed as
In non-small cell lung cancer tumor cell surface；With its ligand binding or with other EGF-R ELISAs (HER2,
EGFR3) formed heterodimer after, can active cell downstream signal transduction approach, lead to the abnormality proliferation of cell.It can be seen that for non-
Small Cell Lung Cancer, EGFR are a very important therapy targets.In particular, in fraction of nonsmall cell lung cancer, such as non-squamous
Cell cancer, there is the 19 exon point mutation of EGFR and the L858R deletion mutations of 21 exons.The tyrosine of EGFR anti-in this way
Kinase inhibitor (tyrosine kinase inhibitor, TKI), such as Gefitinib (Gefitinib), Erlotinib
(Erlotinib), Afatinib (Afatinib), it is especially effective to such patient.Simultaneously relative to chemotherapy, these three medicines
The effective percentage of object is higher, longer without the progression of disease phase, and toxicity religion is small.
But during being treated using this kind of TKI, EGFR can be occurred by carrying the tumour of EGFR sensitizing mutations
The secondary mutation that T790M is shown outside 20, leads to the secondary drug resistance to this kind of TKI, and the incidence of this secondary mutation is about
50%.In addition, also having the mutation that some patientss carry primary EGFRT790M, to this kind of TKI drug resistances.Therefore, in order to overcome
Caused by the secondary and initial drug-resistant of EGFR T790M to Gefitinib (Gefitinib), Erlotinib (Erlotinib), Ah
Method replaces the drug resistance of Buddhist nun (Afatinib), a kind of irreversible EGFR tyrosine kinase inhibitors is had developed, to the T790M of EGFR
The non-small cell lung cancer of mutation, 19 exon point mutation and L858R point mutation has good inhibition, is likely to become Ji Fei
For the second line treatment drug after Buddhist nun (Gefitinib), Erlotinib (Erlotinib), Afatinib (Afatinib) drug resistance.
In the prior art, AZD-9291 is efficient selective EGFR mutant inhibitor；Its molecular formula is C28H33N7O2；Its
Structural formula is shown in formula 1：
AZD9291 is respectively to the IC50 of 19 deletion form EGFR of exon, L858R/T790M EGFR and Wild type EGFR
12.92nM, 11.44nM and 493.8nM.But the half-life period of AZD9291 is shorter, dosage is higher, leads to bad metabolism,
If higher diarrhea, the incidence of fash and nausea increase half-life period therefore, it is necessary to which the structure to the medicine optimizes, with
The dosage of drug is reduced, drug toxicity is reduced.
Invention content
In view of this, the purpose of the present invention is to provide [(indol-3-yl) pyrimidine -2-base] aminophenyl propyl- 2- alkene acyls
Amine derivative and its salt, preparation method, application, [(indol-3-yl) pyrimidine -2-base] aminophenyl propyl- provided by the invention
The half-life period of 2- alkenylamide derivatives increases, and reduces the dosage of drug, reduces drug toxicity.
The present invention provides [(indol-3-yl) pyrimidine -2-base] aminophenyl propyl- 2- alkenylamide derivatives, have Formulas I knot
Structure：
In Formulas I, the R1、R2、R3、R4、R5、R6、R7、R8、R9、R10、R11、R12、R13、R14、R15、R16、R17、R18、R19With
R20It independently is hydrogen or deuterium.
Preferably, the number of D-atom is 9~29 in the Formulas I.
Preferably, the derivative has formula 101,104 structure of formula 102, formula 103 or formula：
The present invention provides the mesylates of derivative described in above-mentioned technical proposal, have Formula II structure：
The present invention provides the preparation methods of derivative described in above-mentioned technical proposal, include the following steps：
Compound with formula III structure and the compound with formula IV structure are reacted, obtained with Formula V structure
Compound；
The compound with Formula V structure is carried out to reduction and amidation successively, obtains the derivative with Formulas I structure
Object；
The R4、R5、R6、R7、R8、R9、R10、R11、R12、R13、R14、R15、R16、R17、R18、R19And R20Independently selected from hydrogen
Or deuterium；
In formula III, the M is F, Cl, Br or I.
Preferably, the compound with formula III structure is made by following preparation method：
Compound with Formula IV structure and the compound with Formula VII structure are reacted, obtained with formula III knot
The compound of structure；
The R11、R12、R13、R14、R15、R16、R17、R18、R19And R20Independently selected from hydrogen or deuterium；
In Formula IV, the R is F, Cl, Br or I.
Preferably, the compound with Formula IV structure and the temperature of the compound reaction with Formula VII structure are 90
DEG C~100 DEG C；Reaction time is 18~22h.
Preferably, the compound with formula III structure and the temperature of the compound reaction with formula IV structure are 93
~98 DEG C；Reaction time is 5.5~6.5h.
The present invention provides a kind of methanesulfonic acids of derivative described in derivative, above-mentioned technical proposal described in above-mentioned technical proposal
The application of derivative prepared by preparation method described in salt or above-mentioned technical proposal in the drug for preparing treatment tumour.
Preferably, the tumour is malignant tumour.
The present invention provides [(indol-3-yl) pyrimidine -2-base] aminophenyl propyl- 2- alkenylamide derivatives, have Formulas I knot
Structure.Deuterium-carbon key in [(indol-3-yl) pyrimidine -2-base] aminophenyl propyl- 2- alkenylamide derivatives provided by the invention makes
Decomposition rate is slow in vivo for it, extends the half-life period of drug, increases the blood concentration of drug, finally reduces it and uses agent
Amount, reduces its toxic side effect.The experimental results showed that：Deuterated derivative AZD 9291-D9 mesylate phases provided by the invention
Than 9291 mesylates of AZD, CmaxIt is 1.32 times of 9291 mesylates of AZD；Its exposed amount (AUC0-t) it is 9291 first of AZD
1.41 times of sulfonate；It eliminates half-life period t1/2It is 1.31 times of 9291 mesylates of AZD, that is, extends 31%,
Description of the drawings
Fig. 1 is the reaction route figure of the embodiment of the present invention 1；
Fig. 2 is the LC-MS detection figures for the brown solid that 1 step 9 of the embodiment of the present invention prepares；
Fig. 3 is the hydrogen nuclear magnetic resonance spectrogram for the brown solid that 1 step 9 of the embodiment of the present invention prepares；
Fig. 4 is the carbon-13 nmr spectra figure for the brown solid that 1 step 9 of the embodiment of the present invention prepares；
Fig. 5 is the LC-MS detection figures for the off-white powder that 1 step 10 of the embodiment of the present invention prepares；
Fig. 6 is the hydrogen nuclear magnetic resonance spectrogram for the off-white powder that 1 step 10 of the embodiment of the present invention prepares；
Fig. 7 is test result of the mesylate drug for preparing of the embodiment of the present invention and comparative example to nude mice；
Fig. 8 is the embodiment of the present invention and the mesylate drug of comparative example preparation to 40~120 days test results of nude mice.
Specific implementation mode
The present invention provides [(indol-3-yl) pyrimidine -2-base] aminophenyl propyl- 2- alkenylamide derivatives, have Formulas I knot
Structure：
In Formulas I, the R1、R2、R3、R4、R5、R6、R7、R8、R9、R10、R11、R12、R13、R14、R15、R16、R17、R18、R19With
R20It independently is hydrogen or deuterium.
In the present invention, the number of D-atom is preferably 9~29 in the Formulas I；In a specific embodiment of the present invention,
In the Formulas I number of D-atom be specially 9,10,11,12,13,14,15,16,17,18,
19,20,21,22,23,24,25,26,27,28 or 29.
In the present invention, the derivative preferably has Formula II, formula III, formula IV or Formula V structure：
The present invention provides a kind of mesylates of derivative described in above-mentioned technical proposal, have Formula II structure：
In the present invention, the number of D-atom is preferably 9~29 in the Formula II；In specific embodiments of the present invention
In, in the Formula II number of D-atom be specially 9,10,11,12,13,14,15,16,17,18
It is a, 19,20,21,22,23,24,25,26,27,28 or 29.
In the present invention, the mesylate of the derivative preferably has formula 201,204 structure of formula 202, formula 203 or formula：
The present invention provides a kind of preparation methods of derivative described in above-mentioned technical proposal, include the following steps：
Compound with formula III structure and the compound with formula IV structure are reacted, obtained with Formula V structure
Compound；
The compound with Formula V structure is carried out to reduction and amidation successively, obtains the derivative with Formulas I structure
Object；
The R4、R5、R6、R7、R8、R9、R10、R11、R12、R13、R14、R15、R16、R17、R18、R19And R20Independently selected from hydrogen
Or deuterium；
In formula III, the M is F, Cl, Br or I.
The present invention reacts the compound with Formula IV structure and the compound with Formula VII structure, is had
The compound of Formula V structure.
In the present invention, the compound with formula III structure is made by following preparation method：
Compound with Formula IV structure and the compound with Formula VII structure are reacted, obtained with formula III knot
The compound of structure；
The R11、R12、R13、R14、R15、R16、R17、R18、R19And R20Independently selected from hydrogen or deuterium；
In Formula IV, the R is F, Cl, Br or I.
The present invention does not have special limitation to the source of the compound with Formula IV structure, using people in the art
The compound with Formula IV structure, such as may be used its commercial goods, can also use people in the art known to member
The technical solution that the compound with Formula IV structure is prepared known to member is voluntarily prepared.In the present invention, described that there is Formula IV knot
The compound of structure is preferably made by the following method：
By compound and three deuterated iodomethane reactions with Formula VII structure, the compound with Formula IV structure is obtained；
The present invention does not have special limitation to the source of the compound with Formula VII structure, using art technology
The compound with Formula VII structure, such as may be used its commercial goods, can also use art technology known to personnel
The technical solution that the compound with Formula VII structure is prepared known to personnel is voluntarily prepared.In the present invention, described to have
The compound of Formula VII structure is preferably made by following preparation method：
The compound with Formula VIII structure and the compound reaction with Formula IX structure are obtained with Formula VII structure
Compound；
In the present invention, the compound with Formula VIII structure is preferably specially shown in formula 301：
In the present invention, the compound with Formula IX structure is preferably specially shown in formula 302：
The present invention in methylene chloride, will preferably be obtained with Formula VIII structure with the dissolving of the compound of Formula VIII structure
Compound solution.The amount of substance and the volume ratio of dichloromethane of the compound with Formula VIII structure are preferably 0.13
~0.16mol：500mL.The present invention is preferably in AlCl3In the presence of carry out Formula VIII structure compound and Formula IX structure change
Close the reaction of object；The AlCl3Amount ratio with the substance of the compound of Formula VIII structure is preferably 0.15~0.2:0.13~
0.15。
In the present invention, the amount ratio of the substance of the compound of the compound and Formula IX structure of the Formula VIII structure is preferably
1:1；The temperature of the compound reaction of the compound and Formula IX structure of Formula VIII structure is preferably 20 DEG C~30 DEG C；Formula VIII structure
Compound and Formula IX structure compound reaction time be preferably 12~15h.The present invention preferably extracts reaction solution.
Present invention preferably employs ethyl acetate EtOAc to be extracted；The number of the extraction is preferably 3 times.
The present invention preferably carries out the reaction of the compound and three deuterated iodomethane with Formula VII structure in a nitrogen atmosphere.
The present invention preferably carries out the anti-of the compound with Formula VII structure and three deuterated iodomethane in the presence of NaH and tetrahydrofuran
It answers.In the present invention, the amount ratio of the substance of the NaH, the compound of Formula VII structure and three deuterated iodomethane be preferably (6.7~
6.8)：(5.5~5.6):(6.6~6.7), more preferably (6.73~6.78)：(5.53~5.55):(6.63~6.68).
In the present invention, the temperature of the compound with Formula VII structure and three deuterated iodomethane reactions is preferably 0 DEG C；It is described to have
The time of the compound of Formula VII structure and three deuterated iodomethane reactions is preferably 1.5 hours.The present invention will preferably have Formula VII
The reaction solution that the compound of structure and three deuterated iodomethane reactions obtain is extracted.Present invention preferably employs ethyl acetate
EtOAc is extracted；The number of the extraction is preferably 3 times.The present invention preferably washs the organic phase being obtained by extraction.This
Invention preferably uses saturated brine to wash；The number of the washing is preferably 3 times.The present invention preferably does washed product
It is dry；Present invention preferably employs anhydrous sulfates to be dried；The anhydrous sulfate is preferably anhydrous sodium sulfate.
In the present invention, the compound with Formula IV structure is preferably specially shown in formula 303：
The present invention does not have the source of the compound with Formula VII structure special limitation, using those skilled in the art
The compound of well known Formula VII structure, such as may be used its commercial goods.In the present invention, described that there is Formula VII structure
Compound preferably be specially formula 304 shown in：
The present invention preferably carries out the compound with Formula IV structure and is had in the presence of p-methyl benzenesulfonic acid and sec-amyl alcohol
The reaction of the compound of Formula VII structure.In the present invention, the compound with Formula IV structure, the change with Formula VII structure
The amount ratio for closing the substance of object and p-methyl benzenesulfonic acid is preferably 1:1：1.15~1.2；The volume and p-methyl benzenesulfonic acid of the sec-amyl alcohol
Substance amount than be preferably (23~26) mL：(1.15~1.2) mol.In the present invention, the chemical combination with Formula IV structure
Object and with Formula VII structure compound reaction temperature be preferably 90 DEG C~100 DEG C, more preferably 95 DEG C；It is described that there is formula
The compound of VI structures and with Formula VII structure compound reaction time be preferably 18~22h, more preferably 20h.This hair
It is bright that the compound with Formula IV structure and the reaction solution obtained by the reaction of the compound with Formula VII structure are preferably cooled to room
Temperature has solid precipitation.Obtained solid is preferably filtered, washed and dried by the present invention successively.The present invention preferably adopts successively
It is washed with sec-amyl alcohol and water.Present invention preferably employs vacuum drying.
In the present invention, the compound with formula III structure is preferably specially shown in formula 305：
The present invention does not have special limitation to the source of the compound with formula IV structure, using people in the art
The compound with formula IV structure is prepared known to member, its commercial goods such as may be used, can also use this field skill
The technical solution that the compound with formula IV structure is prepared known to art personnel is voluntarily prepared.In the present invention, described that there is formula
The preparation method of the compound of IV structures preferably includes following steps：
By with Formula X structure compound and deuterated dimethylamine reaction, obtain the compound with Formula XI structure；
It will be restored with the compound of Formula XI structure, obtain the compound with formula IV structure；
The present invention by with Formula X structure compound and deuterated dimethylamine react, obtain the compound with Formula XI structure.
In the present invention, the compound with Formula X structure is preferably specially shown in formula 306：
In the present invention, the deuterated dimethylamine is preferably participated in the hydrochloride form of deuterated dimethylamine anti-；It is described to have
The amount ratio of the substance of the compound of Formula X structure and deuterated dimethylamine is preferably 34~35：22~23.In the present invention, the tool
The temperature of the compound and the reaction of deuterated dimethylamine that have Formula X structure is preferably 20 DEG C~30 DEG C；The chemical combination with Formula X structure
The time of object and the reaction of deuterated dimethylamine is preferably 20~for 24 hours.
After obtaining the compound with Formula XI structure, the present invention will be restored with the compound of Formula XI structure, be had
The compound of formula IV structure.
The reduction of the compound with formula IV structure is carried out present invention preferably employs tetrahydrochysene lithium aluminium.In the present invention, described
The amount ratio of the substance of tetrahydrochysene lithium aluminium and compound with formula IV structure is preferably 9~11:1, more preferably 10:1.The present invention is excellent
It is selected in the presence of anhydrous tetrahydro furan and carries out above-mentioned reduction.In the present invention, the temperature of the reduction is preferably 0~5 DEG C；Institute
The time for stating reduction is preferably 5.5~6.5h.The present invention is preferably quenched reaction solution, filters, the adjusting of pH value, extraction and
It is dry.Present invention preferably employs water to be quenched.The pH value of reaction solution is preferably adjusted to 2~3 by the present invention.The present invention preferably will
Reaction solution after pH value adjusting and water mixing, then extract, obtain water phase.The present invention is preferably by the water phase using freeze-drying
Mode is dried.
In the present invention, the compound with formula IV structure is preferably specially shown in formula 307：
In the present invention, the reaction of the compound with formula III structure and the compound with formula IV structure is preferred
In N, carried out in the presence of N- diisopropylethylamine (DIEA) and DMAC N,N' dimethyl acetamide (DMAC).In the present invention, described
Compound with formula III structure and the temperature of the compound reaction with formula IV structure are preferably 93~98 DEG C；It is described to have
The compound of formula III structure and with formula IV structure compound reaction time be preferably 5.5~6.5h.In the present invention,
The amount ratio of the substance of the compound with formula III structure and the compound with formula IV structure is preferably 7~9:10.
The present invention is preferably by the compound with formula III structure and the reaction obtained by the reaction of the compound with formula IV structure
Liquid is cooled to 20~30 DEG C.The present invention preferably mixes the reaction solution of cooling and water, and solid is precipitated in stirring.The present invention preferably will
There is the reaction solution that solid is precipitated to filter, obtains filter cake；The filter cake is washed；The filtrate is extracted successively, is washed
It washs, dry and concentrate, obtain solid product；Solid product and filter cake are mixed, the compound with Formula V structure is obtained.
After obtaining the compound with Formula V structure, the present invention restores the compound with Formula V structure and acyl successively
Amination obtains the derivative with Formulas I structure.The present invention preferably carries out the change of Formula V structure in the presence of iron powder and ammonium chloride
Close the reduction of object.The present invention carries out the reduction of the compound of Formula V structure preferably in the mixed solution of second alcohol and water.The present invention is excellent
The compound of Formula V structure is warming up to reflux and reacted by choosing；The time of the compound reduction of the Formula V structure is preferably 2.5
~3.5h, more preferably 3h.The reaction solution that the present invention preferably restores the compound of Formula V structure cools down and filtering；Will
To filtrate concentrated, concentrate is purified, reduzate is obtained.
Reduzate amidation is obtained the derivative with Formulas I structure by the present invention.Present invention preferably employs 3- chlorine propionyl
Chlorine and NaOH carry out the amidation of reduzate.The present invention preferably first reacts reduzate with 3- chlorpromazine chlorides, obtains centre
Body；The intermediate is reacted with NaOH again, obtains the derivative with Formulas I structure.The present invention is preferably in tetrahydrofuran and water
Mixed solvent under carry out amidation.In the present invention, the amount ratio of the substance of the reduzate and 3- chlorpromazine chlorides is preferably
0.8~0.9:1.In the present invention, the amidated temperature is preferably 0 DEG C；The amidated time is preferably 9~11h.
Amidated products are preferably cooled to room temperature by the present invention, then are mixed with water, and mixed liquor is obtained；The mixed liquor is extracted, is obtained
Organic phase；The organic phase is washed, is dried, is concentrated under reduced pressure and purified successively.Present invention preferably employs ethyl acetate into
Row extraction.Present invention preferably employs anhydrous sulfates to be dried.It is 1 present invention preferably employs volume ratio:10 methanol and two
Chloromethanes is purified as eluent.
In the present invention, the mesylate of the derivative has Formula II structure.Mesylate of the present invention to derivative
Preparation method there is no special limitation, using mesylate well known to those skilled in the art at salt mode.At this
In invention, the preparation method of the mesylate of the derivative preferably includes following steps：
Derivative described in above-mentioned technical proposal and methanesulfonic acid are reacted, the mesylate of derivative is obtained.
The derivative is preferably first dissolved in the mixed solution of acetone and water by the present invention, obtains derivative solution.
In the present invention, the amount of the substance of the derivative and the volume ratio of mixed solution are preferably (0.07~0.09) mol：(0.4~
0.5) mL, more preferably 0.08mol：0.44mL；The volume ratio of the acetone and water is preferably 10:1.In the present invention, described
Methanesulfonic acid preferably participates in reacting with the acetone soln of methanesulfonic acid；The volume ratio of the amount and acetone of the methanesulfonic acid and substance is preferably
(0.07~0.09) mmol：(0.045~0.055) mL, more preferably 0.08mmol：0.05mL.
In the present invention, the derivative and the temperature of methanesulfonic acid reaction are 55~65 DEG C；The derivative and methanesulfonic acid
The time of reaction is preferably 1~5 hour.
The present invention preferably carries out derivative and methanesulfonic acid reaction solution obtained by the reaction to be cooled to 20 DEG C~30 DEG C, there is white
Solid is precipitated.The present invention preferably filters cooling reaction solution, is washed, is obtained with methyl tertiary butyl ether(MTBE) (MTBE) to obtained solid
To the mesylate of derivative.
In the present invention, the pharmaceutically acceptable salt of the derivative, crystalline hydrate, solvate, prodrug,
Monocrystalline type object or polymorph are all within the scope of the present invention.
The present invention also provides a kind of methylsulphurs of derivative described in derivative, above-mentioned technical proposal described in above-mentioned technical proposal
The application of derivative prepared by preparation method described in hydrochlorate or above-mentioned technical proposal in the drug for preparing treatment tumour.
In the present invention, the tumour is preferably malignant tumour；The malignant tumour preferably includes medullary carcinoma of thyroid gland, non-
Small Cell Lung Cancer, cancer of pancreas, kidney, metastatic renal cell cancer, carcinoma of urinary bladder, oophoroma, the cancer of the brain, breast cancer prostate cancer, it is multiple
Property myeloma, soft tissue sarcoma, melanoma cancer, recurrence/metastatic Merkel cell cancers, carcinoma of endometrium, plexi nerve it is fine
Tie up tumor, liver cancer, late period cholangiocellular carcinoma, adult's glioblastoma, the straight cancer of metastatic knot or acute myelocytic leukemia.Into
One step is preferably medullary carcinoma of thyroid gland or prostate cancer or liver cancer or kidney.In some embodiments of the invention, targeted
Malignant tumour is non-small cell lung cancer.
In the present invention, the mesylate of the derivative or derivative further includes in the drug as treatment tumour
Pharmaceutically acceptable auxiliary material.In the present invention, the pharmaceutically acceptable auxiliary material includes：Pharmaceutically acceptable carrier,
It is one or more in excipient, diluent, adjuvant, medium.
Can be used as pharmaceutically acceptable carrier substance includes but is not limited to：Sugar, such as lactose, dextrose and saccharose；It forms sediment
Powder such as cornstarch and potato starch；Haemocyanin, such as human albumin；Cellulose and its derivates such as carboxymethyl cellulose
Sodium, ethyl cellulose and cellulose acetate；Buffer substance, such as phosphate, glycine, sorbic acid, potassium sorbate；Ion exchange
Agent, aluminum stearate lecithin；The partial glyceride mixtures of saturated vegetable fatty acid, water, salt or electrolyte, such as sulfuric acid milt egg
In vain, sodium chloride, disodium hydrogen phosphate, polyvinylpyrrolidone, colloidal silicon, potassium hydrogen phosphate, magnesium trisilicate, polyacrylate, zinc salt,
Lanolin, polyethylene-polyoxypropylene-blocking condensate；Gum powder；Malt；Gelatin；Talcum powder；Auxiliary material such as cocoa butter and bolt
Agent wax；Oil such as peanut oil, safflower oil, cotton seed oil, sesame oil, olive oil, soya-bean oil and corn oil；Glycol compound, such as third
Glycol and polyethylene glycol；Esters such as ethyl oleate and ethyl laurate；Agar；Buffer such as aluminium hydroxide and hydrogen-oxygen
Change magnesium；Alginic acid；Pyrogen-free water；Isotonic salt；Lin Ge (family name) solution；Ethyl alcohol, phosphate buffer solution and other are nontoxic suitable
Lubricant such as Sodium Laurylsulfate and magnesium stearate, colorant, releasing agent, coating agents, sweetener, flavoring agent and fragrance are prevented
Rotten agent and antioxidant.
In the present invention, the tumour medicine that the mesylate of the derivative or derivative is prepared can by oral administration to
Medicine, drug administration by injection, Aerosol inhalation administration, local administration, per rectum administration, nose administration, buccal administration, vagina administration or logical
Cross the administration of implantable medicine box.In the present invention, the drug administration by injection includes subcutaneous, vein, intramuscular, intra-articular, sliding
Injection in film (chamber), intrasternal, in film, intraocular, in liver, intralesional, encephalic or infusion techniques.In this hair
In bright, pharmaceutical administration is preferably administered orally, to Intraperitoneal medication or intravenous injection.In the present invention, drug is sterile
Injection system can be water or oil suspension, these suspension can according to known technology using suitable dispersant,
Wetting agent and suspending agent are manufactured by formula.Aseptic injection can be aseptic parenteral solution or suspension, inject nontoxic be subjected to
Diluent or solvent, such as 1,3-BDO solution.These acceptable excipient and solvent can be water, sodium chloride solution and
Ringer's solution etc..Further, sterile non-volatile oil can be used as solvent or suspension media by convention.
The solid dosage forms of oral medication may include tablet, capsule, pulvis, granula and pill.Further, these dosage forms
In, reactive compound is mixed at least one pharmaceutically acceptable inert excipient or carrier or filler, as sodium citrate,
Calcium phosphate.Filler such as lactose, starch, glucose, mannitol, sucrose or silicic acid；Adhesive such as alginates, carboxymethyl cellulose
Element, sucrose, polyvinyl pyrrolidone, gelatin or Arabic gum；Absorbent such as bentonite or white bole；Moisturizer such as glycerine；Disintegrant
Such as calcium carbonate, potato starch, alginic acid, agar, tapioca, certain silicates or sodium carbonate；Sorbefacient such as quaternary ammonium
Compound；Wetting agent such as hexadecanol and glycerin monostearate；Lubricant such as solid polyethylene glycol, magnesium stearate, calcium stearate,
Talcum powder, Sodium Laurylsulfate and their mixture；Block agent solution such as paraffin；As for dosage forms such as tablet, capsule or pills
It may include buffer.
Solid dosage forms may include sugar-pill, tablet, granule, pill or capsule etc., and coating can be used or is prepared by shell material,
Such as casing or other materials well known in the art.And reactive compound can be discharged with the mode of delay in such composition,
Certain part of release in alimentary canal.Further, reactive compound can be the same as one or more shapes in above-mentioned excipient
At microencapsulation form.
Tumour medicine can also be incorporated into suitable delivery vector, such delivery vector can provide control and/or
Compound is continuously discharged, targeting moiety is also can be used as.The example of delivery vector without limitation include adjuvant, synthetic adjuvant,
Microcapsules, particle, liposome and yeast cell wall particle.The wall of yeast cells can be treated differently selectively to remove
Deproteinized matter ingredient, glucan or mannosan layer.
Tumour medicine can be administered orally with any acceptable peroral dosage form, including but be not limited to：
Capsule, tablet, water suspension or water solution.It is administered orally about tablet and enteric coatel tablets dosage form, filler generally comprises breast
Sugar, sucrose, cornstarch, lactose, glucose, silicic acid, sodium citrate, mannitol, Dicalcium Phosphate；Binding such as starch, fiber
Plain derivative, gel, polyvinylpyrrolidone (pvp), sucrose；Lubricant such as glycerine；Disintegrant such as agar, calcium carbonate, horse
Bell potato, tapioca, alginic acid or croscarmellose sodium；Lubricant such as magnesium stearate and glycerine are all typically added
Add.Capsule oral is administered, suitable diluent includes lactose and dry cornstarch.When oral medication is water suspension
When, active ingredient is made of emulsifier and suspending agent.When preparing these dosage forms, pharmaceutically acceptable sweetener, flavoring agent
Or colorant can also be added.
In the present invention, the dosage of the drug for the tumour that the mesylate of the derivative or derivative is prepared is taken an examination
Consider the factors such as administration route, patient health situation.In the present invention, for the people of 60kg weight, day dosage it is preferred
For 1~1000mg, more preferably 5~100mg.
In order to further illustrate the present invention, with reference to embodiment to [(indol-3-yl) pyrimidine -2- provided by the invention
Base] aminophenyl propyl- 2- alkenylamide derivatives and its salt, preparation method, using being described in detail, but cannot be by them
It is interpreted as limiting the scope of the present invention.
Embodiment 1
Reaction route as shown in Figure 1, Fig. 1 are the reaction route figure of the embodiment of the present invention 1：
Step 1：Prepare 3- (2- chlorine pyrimidine-4-yl) -1H- indoles
N2Under atmosphere, AlCl is added portionwise into dichloromethane (500mL) solution of compound 1 (21g, 0.14mol)3
(22.5g, 0.17mol) is finished for 10 minutes.Gained suspension is stirred at room temperature after ten minutes, and reaction system is clarified substantially.Room
Under temperature, compound 2 (16.5g, 0.14mol) is added portionwise into above-mentioned reaction solution, adds within 10 minutes.Gained brown suspension,
Under stirring at room temperature overnight；Reaction is monitored through TLC, and after compound 1 consumes completely, reaction solution is poured into ice water under stirring
In (500mL), this system extracts 3 times through EtOAc (300mL).Merge organic phase, and be washed with brine (200mL) 2 times, through nothing
After aqueous sodium persulfate drying, 38g viscous brown crude products are concentrated under reduced pressure to obtain.The crude product purifies (eluent n- through silica gel column chromatography
hexane:EtOAc=5:1 to 3:1) 16g target compounds 3, yellow solid, yield 49%, are obtained.
The LC-MS test results of compound 3 are：LCMS(ESI+):m/z 230.15(M+H)；1H-NMR(300MHz,
DMSO-d6) δ 12.09 (brs, 1H), 8.59-8.54 (m, 2H), 8.44-8.40 (m, 1H), 7.92 (d, J=5.4Hz, 1H),
7.52-7.50(m,1H),7.34-7.20(m,2H)。
Step 2：Prepare three deuterated Methvl-indoles of 3- (2- chlorine pyrimidine-4-yl) -1-
Under nitrogen atmosphere, under ice bath cooling, to 3- (2- chlorine pyrimidine-4-yl) -1H- indoles 3 (1.27g, 5.53mmol)
In THF (50mL) solution, 60%NaH (0.27g, 6.75mmol) is added portionwise, 20min is finished, this reaction solution is stirred at 0 DEG C
0.5 hour, three deuterated iodomethane (0.96g, 6.64mmol) are added dropwise into above-mentioned reaction system, 10min drops finish, and anti-at 0 DEG C
It answers 1.5 hours, TLC monitoring reactions；When raw material 3 is exhausted, reaction terminates, and saturated sodium bicarbonate is slowly added dropwise into reaction solution
Reaction is quenched in solution (10mL).Gained mixture EtOAc (40mL) is extracted 3 times, merges organic phase, saturated salt is used in combination to wash
Wash (40mL) 3 times；After anhydrous sodium sulfate drying, 1.29g yellow solids 4 are concentrated under reduced pressure to obtain, yield 94.9%, the compound 4 is not
Through further purifying, it is directly used in next step.
The test result of compound 4 is：HPLC:96.8%；LCMS(ESI+):m/z 247.16(M+H)；1H-NMR
(300MHz,CDCl3)δ8.46-8.42(m,1H),8.38-8.31(m,1H),7.95(s,1H),7.50-7.46(m,1H),
7.40-7.28(m,3H)。
Step 3：It is phonetic to prepare N- (the fluoro- 2- methoxyl groups -5- nitrobenzophenones of 4-) -4- (tri- deuterated Methvl-indole -3- of I- certain)
Pyridine -2- amine
At room temperature by compound 4 (1.19g, 4.82mmol), compound 5 (0.90g, 4.82mmol) and p-TsOH.H2O
(1.1g, 5.72mmol) is added in sec-amyl alcohol (24mL), and gained suspension is replaced 3 times through nitrogen, and stirring after five minutes, is warming up to
95 DEG C of reactions.HPLC monitoring reactions.After 20 hours, HPLC shows that raw material compound 4 completely disappears, and reaction terminates.By reaction solution
It is cooled to room temperature, stirs 1 hour, there is solid precipitation.It filters, filter cake is washed with sec-amyl alcohol (2mL) and water (2mL) successively.Gained
Solid is vacuum dried to obtain 1.8g yellow solid compounds 6.The product is directly used in next step without further purifying.
The test result of compound 6 is：HPLC:94.8% (deducting p-TsOH)；LCMS(ESI+):m/z 397.11(M+
H)；1H-NMR (300MHz, DMSO-d6) δ 9.84 (brs, 1H), 8.75 (d, J=8.1Hz, 1H), 8.69 (t, J=2.1Hz,
1H), 8.32 (d, J=6.6Hz, 1H), 8.20 (d, J=7.2Hz, 1H), 7.59 (t, J=8.1Hz, 1H), 7.53-7.47 (m,
2H), (crude product contains about 25% by 7.31 (t, J=7.5Hz, 1H), 7.13 (t, J=8.1Hz, 1H), 4.00 (s, 3H)
TsOH)。
Step 4：Prepare intermediate 7-2
It is water-soluble that 7 (10g, 0.13mol, 1.0eq), 50mL water and 50mL (1mol/L) sodium hydroxide are added into reaction bulb
Liquid.It is cooled to 0~5 DEG C with ice-water bath after stirring clarification, the toluene solution of 7-1 (14.4g, 0.13mol, 1.0eq) is then added dropwise
(30mL) supplements sodium hydrate aqueous solution (1M, 216mL) simultaneously during being added dropwise, system pH is made to be maintained between 8~10.
Room temperature is moved to after adding and is stirred 30 minutes.500mL water dilute reaction solutions are added, are extracted (250mL x 3) with EtOAc.It receives
The dilute hydrochloric acid (1mol/L) of collection water phase is adjusted to PH to 3~4, and with extraction, EtOAc extracts (250mL x three times to water phase again
3).EtOAc phases Na2SO4Dry, filtering and concentrating obtains compound 7-2.
1H NMR(300MHz,CDCl3)δ5.28(s,2H),4.20-4.15(m,2H),4.13-4.02(m,2H),1.27
(t, 3H, J=7.2Hz).
Step 5：Prepare intermediate 7-3
At room temperature, to the chlorine of Pvi-Cl (4.13g, 34.27mmol, 1.5eq) and TEA (4.0g, 39.53mmol, 1.7eq)
In imitative (55mL) solution, the chloroformic solution (55mL) of compound 7-2 (5.04g, 34.27mmol, 1.5eq) is slowly added dropwise.It is added dropwise
After, then under nitrogen protection, be stirred at room temperature 1.5 hours.Under nitrogen protection, be added deuterated dimethylamine hydrochloride (2.0g,
22.84mmol, 1.0eq), it is then stirred at room temperature under enclosed system 22 hours.100mL water is added, it is small to be stirred at room temperature 1
When, 40mL chloroforms are added, organic phase is primary with 100mL water washings, then is washed with the aqueous sodium carbonate (1mol/L) of 100mL
3 times, then it is primary with the water washing of 100mL.Chloroform mutually uses Na2SO4Dry, filtering crosses column purification, obtains compound 7-3 after concentration
(3.0g, yield：72%)；
1H NMR(300MHz,CDCl3)δ5.68(s,1H),4.21-4.10(m,2H),4.02-3.98(m,2H),1.27
(t, 3H, J=7.2Hz)
Step 6：Prepare N- methyl-N ', N '-two (three deuterated methyl) 1,2- ethylenediamines 8
Compound 7-3 (3.0g, 16.6mmol, 1.0eq) is dissolved in the anhydrous THF of 100mL.Ice water is used under nitrogen protection
Bath is cooled to 0~5 DEG C, is then slowly added to tetrahydrochysene lithium aluminium (7.0g, 166mmol, 10eq) in batches, waits stablizing no bubble formation
Afterwards, it is slowly ramped to micro- reflux.Reaction terminates after 6 hours.System is cooled to room temperature, and is cooled to 0~5 DEG C with ice-water bath, dropwise
It is slowly added to 7mL water quenchings to go out, then is slowly added to the sodium hydrate aqueous solution of 7mL 20% dropwise, then be slowly added to 7mL water dropwise.
After being stirred at room temperature 30 minutes, filtering is used in combination THF washings filter cake three times, collects filtrate, is adjusted solution after pH 2-3 with dilute hydrochloric acid
It is evaporated, adds water (150mL), three times with EtOAc (150mL x 3) extractions.The chilled freeze-drying of water phase is dry, obtains compound 8
(2.6g, yield：86%).
The test result of compound 8 is：1H NMR(300MHz,D2O)δ3.51-3.42(m,4H),2.74(s,3H)。
Step 7：Prepare N '-(2- [two (three deuterated methyl) amino] ethyl) -2- methoxyl groups-N '-methyl-N- [4- (1-
Three deuterium methyl draw diindyl -3- bases) pyrimidine -2-base] -5- nitre benzene -1,4- diamines 9
At room temperature, by compound 6 (300mg, 0.76mmol), compound 8 (170.0mg, 0.94mmol) and DIEA
(440.0mg, 3.4mmol) is added in DMAC (3mL), and 95 DEG C of reactions are warming up in vexed tank, and HPLC monitoring is reacted, after 6 hours,
HPLC shows that raw material 6 completely disappears, and reaction terminates；Reaction solution is down to room temperature, water (3mL) is added, is stirred at room temperature 2 hours, has
Solid is precipitated, and filters, and filter cake is washed with water (2mL)；Filtrate is extracted 3 times with EtOAc (30mL), merges organic phase, and be washed with water
6 times (30mL) are washed to remove DMAC.It is dried over anhydrous sodium sulfate, brick-red solid compound 9, yield is obtained after reduced pressure
73%.The product is directly used in next step without further purifying.
The test result of compound 9 is：LCMS(ESI+):m/z 485.33(M+H)；1H-NMR (crude product 300MHz,
CDCl3) δ 9.59 (s, 1H), 8.41-8.18 (m, 4H), 7.63 (s, 1H), 7.41-7.20 (m, 2H), 7.20 (d, J=5.1Hz,
1H), 6.82 (s, 1H), 4.04 (s, 3H), 3.30 (t, J=6.9Hz, 2H), 3.0 (s, 3H), 2.60 (t, J=6.9Hz, 2H).
Step 8：Prepare N1(2- [two (three deuterated methyl) amino] ethyl) -5- methoxyl groups-N1Methyl-N4-[4-(1-
Three deuterium methyl draw diindyl -3- bases) pyrimidine -2-base] benzene -1,2,4- triamines 10
At room temperature by compound 9 (1.08g, 2.23mmol), iron powder (0.750g, 13.38mmol) and NH4Cl
(0.084g, 1.57mmol) adds to EtOH/H2In O (60mL/20mL), and it is warming up to reflux.After 3 hours, TLC shows raw material 9
It completely disappears, reaction terminates；Reaction solution is down to room temperature, is filtered；Filter cake is washed 3 times with ethyl alcohol (10mL), filtrate decompression concentration,
Gained residue is through silica gel column chromatography (eluent MeOH:DCM=1:10,1% ammonium hydroxide) purification, obtain 790mg beige solids
Close object 10, yield 78%.
The test result of compound 10 is：HPLC:98.1%；LCMS(ESI+):m/z 455.33(M+H)；1H-NMR
(300MHz,CDCl3) δ 8.48-8.46 (m, 1H), 8.34 (d, J=5.1Hz, 1H), 8.20 (s, 1H), 7.82 (s, 1H), 7.64
(s, 1H), 7.39-7.28 (m, 3H), 7.05 (d, J=5.1Hz, 1H), 6.67 (s, 1H), 3.86 (s, 3H), 3.49 (s, 2H),
3.30 (t, J=5.7Hz, 2H), 3.08 (d, J=5.7Hz, 2H), 2.72 (s, 3H)
Step 9：Prepare N- (2- { 2- [two (three deuterated methyl) amino] ethyl-(methyl) amino } -4- methoxyl groups -5-
{ [4- (tri- deuterated methyl indol -3- bases of 1-) pyrimidine -2-base] amino } phenyl) propyl- 2- acrylamides (AZD 9291-d9)
Under ice bath cooling, to the THF/H of compound 10 (0.7g, 1.54mmol)23- is added dropwise in O (6mL/0.6mL) solution
Chlorpromazine chloride (0.24g, 1.85mmoL), drop finishes within 10 minutes；This reaction solution continues to stir at 0 DEG C.After 1 hour, TLC and LCMS
Display raw material 10 is exhausted；NaOH (0.25g, 6.15mmoL) is added in above-mentioned reaction solution, and is warming up to reflux.9 hours
Afterwards, LCMS and TLC shows that intermediate fundamental reaction finishes, and room temperature is down in reaction, water (1mL) is added, EtOAc is used in combination
(30mL) is extracted 3 times, is merged organic phase, (10mL) is washed with water 3 times.It is dried over anhydrous sodium sulfate, 0.9g palm fibres is concentrated under reduced pressure to obtain
Color dope crude product.The crude product is through silica gel column chromatography (eluent MeOH:DCM=1:10) it purifies, it is solid to obtain 0.61g foam-like browns
Body compound 11, yield 78%.
The present invention carries out LC-MS detections to the compound that step 9 obtains, and testing result is as shown in Fig. 2, Fig. 2 is the present invention
The LC-MS detection figures for the brown solid that 1 step 9 of embodiment prepares, as seen from Figure 2：LCMS(ESI+):m/z
509.30(M+H)；
The present invention carries out nuclear magnetic resonance spectroscopy detection to the compound that step 9 obtains, and testing result is as shown in figure 3, Fig. 3
For the hydrogen nuclear magnetic resonance spectrogram for the brown solid that 1 step 9 of the embodiment of the present invention prepares, as seen from Figure 3：1H-NMR
(300MHz,CDCl3) δ 10.21 (brs, 1H), 9.87 (s, 1H), 9.14 (s, 1H), 8.40 (d, J=5.4Hz, 1H), 8.09-
8.07 (m, 1H), 7.74 (s, 1H), 7.43-7.40 (m, 1H), 7.32-7.23 (m, 2H), 7.22 (d, J=5.4Hz, 1H),
6.82(s,1H),6.52-6.35(m,2H),5.72(dd,J1=2.4Hz, J2=9.6Hz, 1H), 3.96 (s, 3H), 2.91 (t, J
=5.4Hz, 2H), 2.73 (s, 3H), 2.30-2.28 (m, 2H).
The present invention carries out carbon-13 nmr spectra test to the compound that step 9 obtains, and test result is as described in Figure 4, Fig. 4
For the carbon-13 nmr spectra figure for the brown solid that 1 step 9 of the embodiment of the present invention prepares.
Step 10：Prepare N- (2- { 2- [two (three deuterated methyl) amino] ethyl-(methyl) amino } -4- methoxyl groups -5-
{ [4- (tri- deuterated methyl indol -3- bases of 1-) pyrimidine -2-base] amino } phenyl) propyl- 2- acrylamides mesylate (AZD 9291-
D9 mesylates)
Compound 11 (40mg, 0.08mmol) is dissolved in acetone/water (0.4mL/0.04mL), and is warming up to 60 DEG C, once
Property be added methanesulfonic acid (7.6mg, 0.08mmoL) acetone (0.05mL) solution, this reaction solution continue at 60 DEG C stirring 1.5 hours
Cooled to room temperature afterwards has off-white powder precipitation, and filtering is washed with MTBE (1mL), dry 44.6mg white solids 12
(AZD 9291-d9 mesylates)；The yield of off-white powder AZD 9291-d9 mesylates is 94%.
Chemical purity>98%, deuterated purity>99%；
The present invention carries out LC-MS detections to the off-white powder that 1 step 10 of embodiment obtains, and detection figure is as shown in figure 5, figure
The LC-MS detection figures of the 5 off-white powder AZD 9291-d9 mesylates prepared for 1 step 10 of the embodiment of the present invention, from
Fig. 5 can be seen that：LCMS(ESI+):m/z 509(M-OMs)；
The present invention carries out nuclear magnetic resonance spectroscopy test to the off-white powder that step 10 obtains, and test results are shown in figure 6,
Fig. 6 is the nuclear magnetic resonance spectroscopy for the off-white powder AZD 9291-d9 mesylates that 1 step 10 of the embodiment of the present invention prepares
Figure.
As can be seen from Figure 6：1H-NMR(300MHz,DMSO-d6)δ9.60(s,1H),9.18(s,1H),8.78(s,
1H), 8.54 (s, 1H), 8.32 (d, J=5.7Hz, 2H), 7.96 (s, 1H), 7.53 (d, J=8.1Hz, 1H), 7.27-7.13
(m,3H),7.02(s,1H),6.69(dd,J1=9.9Hz, J2=16.5Hz, 1H), 6.33 (d, J=16.5Hz, 1H), 5.82
(d, J=10.2Hz, 1H), 3.91 (s, 3H), 3.28 (brs, 4H), 2.63 (s, 3H), 2.09 (s, 3H).
The experiment in vitro of 1.1 deuterated derivative AZD 9291-D9 mesylates
The AZD 9291-D9 mesylates that Example 1 prepares investigate it to PC-9 and PC-9 persister cells
Lethal effect, using 9291 mesylates of AZD and Gefitinib as positive control, PBS is as blank control.
Respectively by each 20000 of two kinds of cells of PC-9 and PC-9 persisters, it is inoculated into 7 75cm2Tissue Culture Flask carries out
Cell culture.By cell secondary culture in 37 DEG C, 5% carbon dioxide incubator counts, and 1000rpm is centrifuged 5 minutes, with culture
The whole cell density of keynote, according to 2000/100 holes μ L/, the inoculating cell in 96 orifice plates；Second day, difference agent-feeding treatment, 100
μ g/mL, 30 μ g/mL, 10 μ g/mL, 3 μ g/mL, 1 μ g/mL, 0.3 μ g/mL, 0.1 μ g/mL, 0.03 μ g/mL, 0.01 μ g/mL, root
According to above-mentioned cell doubling time, respectively the PC-9 cells after dosing cultivate within 3 days, PC-9 persisters cultivate within 5 days.
Day is detected, the CCK8 reagents (Japanese colleague's chemistry) of 100 μ L are added into each cell culture well, it is small to be put into incubator standing 1
When, light absorption value is measured in 450nm with Beijing Pu Lang companies microplate reader (model DNM9602), using GraphPad softwares, is calculated
IC50, sample to be tested are shown in Table 1 to PC-9 and PC-9 persister lethal effect testing results；And Gefitinib is to PC-9 drug resistances
Strain lethal effect testing result, is shown in Table 2：
1 sample to be tested of table is to PC-9 and PC-9 persister lethal effect testing results
2 Gefitinib of table is to PC-9 persister lethal effect testing results
As known from Table 1, experimental group and positive controls sample to be tested are suitable to the lethal effect of PC-9 persister cells, raw
Long half-inhibition concentration IC50Quite, difference is not notable, (P ＞ 0.05), illustrates the deuterated AZD that the embodiment of the present invention 1 prepares
9291-D9 mesylates are suitable with the lethal effect to cancer cell of 9291 mesylates of AZD, i.e., by 9291 first of AZD
After sulfonate is deuterated, its Bioactivity will not be changed.
The Pharmacokinetic Evaluation of 1.2 deuterated derivative AZD 9291-D9
The deuterated AZD 9291-D9 mesylates that Example 1 prepares, pharmacokinetic studies are carried out to it, with
9291 mesylates of AZD are as positive control, and PBS is as blank control.
Animal subject is：Male CD1 mouse, 18~22g of weight, the animal center tested by the drug of Shanghai provide, and use
Credit number SYXK (Shanghai) 2010-0049.Animal subject should carry out adaptability raising in 3~7 days a few days ago in experiment in test site,
Male CD1 mouse, are randomly divided into 2 groups, and gavage is given by test agent respectively, fasting 12 hours before testing, free water, administration 2
Unified feed after hour；Single oral gavage gives the sample to be tested of 25mg/kg dosage, the sample to be tested of experimental group and positive controls
Use 0.5%w/v hydroxypropyl methyl celluloses (HPMC)；
The aqueous dissolution of 0.1%w/v tweens；Distinguish 0.25h, 0.5h, 1.0h, 2.0h, 3.0h, 5.0h upon administration,
It 8.0h, 10h and samples for 24 hours；3 mouse of each time point, it is quiet after eyeball of mouse after the above setting time point Animal Anesthesia
Arteries and veins clump extracting vein blood 0.3mL, sets in heparinised tubes, and 11000g is centrifuged 5 minutes, and separated plasma freezes in -20 DEG C of refrigerators；
When sample detection, plasma sample measures 9291 methylsulphurs of compound AZD in blood plasma after methanol extraction albumen using LC-MS/MS methods
The concentration of hydrochlorate and deuterated AZD 9291-D9 mesylates, the range of linearity are 30.0~30000ng/mL.
Main pharmacokinetic parameters (T after intragastric administration on mice administration is calculated using 6.3 softwares of WinNonlinmax, Cmax, AUC,
MRT and t1/2).Wherein, Cmax C is reachedmaxWith peak time TmaxFor measured value.
Lower area of blood concentration-time curve AUC0-tValue：It is calculated using trapezoidal method.
AUC0-∞=AUC0-t+Ct/ke,
CtThe blood concentration at time point, k can be measured for the last oneeFor elimination rate constant；
Eliminate half-life period t1/2=0.693/ke；
Mean residence time MRT=AUMC/AUC.
Mouse is after gavage gives 9291 mesylate of 25mg/kg AZD 9291-D9 mesylates and AZD respectively, drug
Pharmacokinetic parameters be shown in Table 3, table 3 is that mouse distinguishes the medicine kinetic parameter after gavage 25mg/kg drugs：
3 mouse of table distinguishes the medicine kinetic parameter after gavage 25mg/kg drugs
The experimental data from table 3 is it is found that the deuterated AZD 9291-D9 first that the preparation method that embodiment 1 provides prepares
The C of sulfonatemaxIt is 1.32 times of 9291 mesylates of AZD；Its exposed amount (AUC0-t) it is the 1.41 of 9291 mesylates of AZD
Times；It eliminates half-life period t1/2It is 1.31 times of 9291 mesylates of AZD, that is, extends 31%, illustrates that the present invention prepares
Deuterated AZD 9291-D9 mesylates, compare 9291 mesylates of AZD, significantly improve blood concentration, extend drug
Half-life period, significant difference.
Embodiment 2
Prepare N- (2- { 2- [two (three deuterated methyl) amino] ethyl-(methyl) amino } -4- methoxyl groups -5- { [4- (1- tri-
Five deuterated indol-3-yls of deuterated methyl -2,4,5,6,7-) pyrimidine -2-base] amino } phenyl) propyl- 2- acrylamide mesylates
(AZD 9291-D14 mesylates)
The present embodiment 2 is prepared according to the method described in example 1, and difference is：With 1H- indoles -2,3,4,5,6,7-
D6 replaces 1H- indoles to which target product be made.
After tested：Chemical purity>98%, deuterated purity>98%；LCMS(ESI+):m/z 514(M-OMs).
Comparative example 1
Prepare N- (2- { 2- [two (three deuterated methyl) amino] ethyl-(methyl) amino } -4- methoxyl groups -5- { [4- (1- first
Base indol-3-yl) pyrimidine -2-base] amino } phenyl) propyl- 2- acrylamides mesylate (AZD 9291-D6 mesylates)：
It prepares by the method described in example 1, difference is：Three deuterated iodomethane are replaced with iodomethane, to be made
Target product.
Chemical purity>98%, deuterated purity>99%；LCMS(ESI+):m/z 506(M-OMs).
Comparative example 2
Preparation N- (2- { 2- dimethyl aminoethyls-(methyl) amino } -4- methoxyl groups -5- [4- (methyl -2,4,5,6,
Five deuterated indoles indol-3-yls of 7-) pyrimidine -2-base] amino } phenyl) propyl- 2- acrylamides mesylate (AZD 9291-D5 methylsulphurs
Hydrochlorate)：
It prepares by the method described in example 1, difference is：Three deuterated iodomethane are replaced with iodomethane, with N- methyl-
N ', N '-dimethyl -1,2- ethylenediamine replaces N- methyl-N ', N '-two (three deuterated methyl) 1,2- ethylenediamines, with indoles -2 1H-,
3,4,5,6,7-d6 replaces 1H- indoles to which target product be made.
Chemical purity>98%, deuterated purity>99%；LCMS(ESI+):m/z 508(M-OMs).
Comparative example 3
Prepare N- (2- { 2- dimethyl aminoethyls-(methyl) amino } -4- methoxyl groups -5- { [4- (tri- deuterated methyl Yin of 1-
Diindyl -3- bases) pyrimidine -2-base] amino } phenyl) propyl- 2- acrylamides mesylate (AZD 9291-D3 mesylates)：
It prepares by the method described in example 1, difference is：With N- methyl-N ', N '-dimethyl -1,2- ethylenediamines replace
N- methyl-N ', N '-two (three deuterated methyl) 1,2- ethylenediamines 8 are changed to which target product be made.
Chemical purity>98%, deuterated purity>99%；LCMS(ESI+):m/z 503(M-OMs).
The experiment of nude mouse of the mesylate of 3 deuterated derivative of embodiment：
PC-9 lung carcinoma cells RPMI1640 medium cultures, take 5 × 106(volume：0.1mL) cell inoculation is in BALB/
C-NU nude mice by subcutaneous, and grow 4 months；The case where growth of transplanted human, measures every 2 weeks, and calculates gross tumor volume, works as transplanting
Knurl product grows to 0.2~0.4cm3When, mouse is random according to tumor size progress, it is divided into treatment group and placebo pair respectively
According to group.Intragastrically is administered daily/placebo, measures the mean change of the volume of tumour, is counted with bilateral T inspections.
The AZD9291-D3 prepared respectively with the AZD9291 of 2mg/kg weight, comparative example 1, structure Transplanted tumor model compares
The AZD9291-D9 of AZD9291-D6, the preparation of embodiment 1 prepared by AZD9291-D5, the comparative example 1 of the preparation of example 2, the system of embodiment 2
Standby AZD9291-D14 and PBS negative control gavage feedings, every group 6, the volume of point measurement tumour, swells in different times
The volume unit of tumor is cm3。
7 groups of animals, PC-9 cell nude mice models are set：Blank group, AZD9291 mesylates, AZD9291-D3 methanesulfonic acids
Salt, AZD9291-D5 mesylates, AZD9291-D6 mesylates, AZD9291-D9 mesylates and AZD9291-D14 methylsulphurs
Hydrochlorate drug concentration is 2mg/kg.
To nude mice, test results are shown in figure 7, and Fig. 7 is the embodiment of the present invention and mesylate drug prepared by comparative example
To the test result of nude mice；Wherein,It is blank group to the test curve of nude mice,It is AZD9291 mesylates to nude mice
Test curve,It is AZD9291-D5 mesylates to the test curve of nude mice,Survey for AZD9291-D3 to nude mice
Curve is tried,It is AZD9291-D6 mesylates to the test curve of nude mice,It is AZD9291-D9 mesylates to nude mice
Test curve,It is AZD9291-D14 mesylates to the test curve of nude mice.Fig. 8 is the amplification of partial trace in Fig. 7
Figure, Fig. 8 are the embodiment of the present invention and the mesylate of comparative example preparation to 40~120 days test results of nude mice, wherein
It is AZD9291 mesylates to the test curve of nude mice,It is AZD9291-D3 to the test curve of nude mice,For
AZD9291-D6 mesylates to the test curve of nude mice,It is AZD9291-D9 mesylates to the test curve of nude mice,It is AZD9291-D14 mesylates to the test curve of nude mice,Survey for AZD9291-D5 mesylates to nude mice
Try curve.
To the rejection ability of tumor bearing nude mice tumour, AZD9291-D14=known to experimental data from Fig. 7 and Fig. 8
AZD9291-D9>AZD9291-D6>AZD9291-D3>AZD9291-D5>AZD9291>Blank group；AZD9291-D9 is
1.5 times of AZD9291-D6 are 1.8 times of AZD9291-D3；It is 2 times of AZD9291.In contrast, AZD9291-D14 and
AZD9291-D9 is most strong to the rejection ability of tumor bearing nude mice tumour.
The mesylate of deuterated derivative provided by the invention considerably increases the half-life period of drug, extends drug in people
The time being detained in body body；The concentration for improving drug in blood simultaneously, to reach more preferable curative effect；Compared to 9291 methylsulphurs of AZD
Hydrochlorate, the dosage smaller of the mesylate of deuterated derivative provided by the invention, and then can further eliminate drug not
Good metabolic problems reduce drug toxicity and other side effects.
As seen from the above embodiment, the present invention provides [(indol-3-yl) pyrimidine -2-base] aminophenyl propyl- 2- alkene acyls
Amine derivative has Formulas I structure.[(indol-3-yl) pyrimidine -2-base] aminophenyl propyl- 2- acrylamides provided by the invention are spread out
Deuterium-carbon key in biology makes it, and decomposition rate is slow in vivo, extends the half-life period of drug, and the blood medicine for increasing drug is dense
Degree, finally reduces its dosage, reduces its toxic side effect.
In addition, derivative provided by the invention is more stablized in vivo, reduce it in inner wall of intestine and in liver
Metabolism reduces the bad metabolism of drug, further decreases drug toxicity and other side effects.
The above is only a preferred embodiment of the present invention, it is noted that for the ordinary skill people of the art
For member, various improvements and modifications may be made without departing from the principle of the present invention, these improvements and modifications are also answered
It is considered as protection scope of the present invention.
Claims (6)
1. [(indol-3-yl) pyrimidine -2-base] aminophenyl propyl- 2- alkenylamide derivatives have 102 structure of formula 101 or formula：
2. the mesylate of derivative described in claim 1 has 202 structure of formula 201 or formula：
3. the preparation method of derivative described in claim 1, includes the following steps：
Compound with formula 305 or formula 305-1 structures and the compound with 307 structure of formula are reacted, had
Compound 9 or compound 9-1；
By described there is compound 9 or compound 9-1 to carry out reduction and amidation successively, obtains having spreading out for formula 101 or formula 102
Biology；
4. preparation method according to claim 3, which is characterized in that the chemical combination with formula 305 or formula 305-1 structures
Object and with 307 structure of formula compound reaction temperature be 93~98 DEG C；Reaction time is 5.5~6.5h.
5. the mesylate of derivative or claim 3~4 described in derivative, claim 2 described in a kind of claim 1 are arbitrary
The application of derivative prepared by one preparation method in the drug for preparing treatment tumour.
6. application according to claim 5, which is characterized in that the tumour is malignant tumour.
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| CN111170998B (en) | 2014-11-05 | 2023-04-11 | 益方生物科技(上海)股份有限公司 | Pyrimidine or pyridine compound, preparation method and medical application thereof |
| CA3009850C (en) | 2015-12-27 | 2020-05-26 | NeuForm Pharmaceuticals, Inc. | Deuterated compounds for treating cancer and related diseases and conditions, and compositions and methods thereof |
| CN106995435B (en) * | 2016-01-22 | 2018-06-08 | 焦玉奇 | 2- (2,4,5- substituted anilines) pyrimidine derivatives |
| CN107043369A (en) * | 2016-02-06 | 2017-08-15 | 焦玉奇 | 2 (2,4,5 substituted aniline) pyrimidine derivatives |
| CN109311858B (en) | 2016-05-26 | 2021-12-03 | 里科瑞尔姆Ip控股有限责任公司 | EGFR inhibitor compounds |
| CN107522690B (en) * | 2016-06-20 | 2022-08-05 | 海南先声药业有限公司 | Preparation method of Osimetinib |
| CN108047205B (en) * | 2016-12-14 | 2019-08-27 | 河南真实生物科技有限公司 | 2- (2,4,5- replace phenylamino) pyrimidine derivatives, preparation method and its application in preparation of anti-tumor drugs |
| CN107192773B (en) * | 2017-05-17 | 2019-09-27 | 张家港威胜生物医药有限公司 | A kind of detection is difficult to understand uncommon for Buddhist nun's content and the high-efficient liquid phase technique in relation to substance |
| EP3647312B1 (en) | 2017-05-24 | 2023-05-10 | TYK Medicines Inc. | Crystal form of deuterated azd9291, preparation method therefor, and use thereof |
| CN108675993A (en) * | 2018-06-14 | 2018-10-19 | 纽瑞森生物科技（北京）有限公司 | Deuterated pyrimidines, preparation method, pharmaceutical composition, preparation and purposes |
| CN110950847B (en) * | 2018-09-27 | 2022-11-01 | 浙江同源康医药股份有限公司 | Novel crystal form of deuterated AZD9291 compound and application thereof |
| CN111285852A (en) * | 2020-04-02 | 2020-06-16 | 广州博济医药生物技术股份有限公司 | Crystal form of deuterated oxitinib medicinal salt and preparation method thereof |
| CN111362924B (en) * | 2020-04-21 | 2021-05-28 | 南京雷正医药科技有限公司 | Deuterated pyrimidine derivatives and uses thereof |
| CN111909135A (en) * | 2020-08-15 | 2020-11-10 | 天津大学 | AZD9291 deutero-mesylate new salt form and preparation method thereof |
| CN113582976B (en) * | 2021-08-24 | 2023-03-17 | 郑州大学 | Deuterated 2-substituted aniline-4-indolyl pyrimidine derivative and preparation method and application thereof |
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Inventor after: Zhang Huizhen Inventor after: Peng Kuai Inventor after: Zheng Feiming Inventor after: Fu Yong Inventor before: Peng Kuai Inventor before: Zheng Feiming Inventor before: Fu Yong |