CN106146516B - The preparation method of Buddhist nun's impurity of the drug is replaced according to Shandong - Google Patents
The preparation method of Buddhist nun's impurity of the drug is replaced according to Shandong Download PDFInfo
- Publication number
- CN106146516B CN106146516B CN201510188801.XA CN201510188801A CN106146516B CN 106146516 B CN106146516 B CN 106146516B CN 201510188801 A CN201510188801 A CN 201510188801A CN 106146516 B CN106146516 B CN 106146516B
- Authority
- CN
- China
- Prior art keywords
- impurity
- phenoxyphenyl
- pyrimidine
- shandong
- compound
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D487/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00
- C07D487/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00 in which the condensed system contains two hetero rings
- C07D487/04—Ortho-condensed systems
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D487/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00
- C07D487/12—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00 in which the condensed system contains three hetero rings
- C07D487/14—Ortho-condensed systems
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D519/00—Heterocyclic compounds containing more than one system of two or more relevant hetero rings condensed among themselves or condensed with a common carbocyclic ring system not provided for in groups C07D453/00 or C07D455/00
Abstract
The invention belongs to pharmaceutical synthesis fields, the impurity and preparation method thereof being related in production of raw medicine technique, in particular to a kind of process impurity and preparation method thereof that Buddhist nun 1- [(3R) -3- [4- amino -3- (4- Phenoxyphenyl) -1H pyrazolo [3,4-d] pyrimidine -1- base] -1- piperidyl] -2- propylene -1- ketone is replaced according to Shandong.
Description
Technical field
The invention belongs to pharmaceutical synthesis fields, are related to bulk pharmaceutical chemicals according to Shandong for process impurity and its preparation in Buddhist nun's production process.
In particular to according to Shandong for Buddhist nun's process impurity and preparation method thereof.
Background technique
Ibrutinib was developed by Celera Genomics company of the U.S. in 2007 first, after transfer California
Subsidiary's Janssen Pharmaceutica (Jassen) of the exploitation of Pharmacyclics company, Johson & Johnson in 2011 participates in cooperative development.At present
It there is no the Chinese translation of standard, therefore its transliteration is herein " replacing Buddhist nun according to Shandong " by the applicant.The medicine is obtained in November, 2013
FDA approval, for previously receiving lymphoma mantle cell (MCL) patient of lenalidomide at least once or other drugs treatment
Treatment.According to Shandong for Buddhist nun be it is first once a day, single formulation, oral bruton's tyrosine kinase (BTK) inhibitor.2014 2
12 Yi Lu of the moon is ratified to increase indication by FDA for Buddhist nun, for treating chronic lymphocytic leukemia (CLL).July 28 in 2014
Day, FDA is announced again, is expanded approval according to Shandong and is replaced Buddhist nun's indication, is used for the white blood of chronic lymphocytic of chromosome 17P missing
Sick (CLL) patient.Treatment Waldenstrom macroglobulinemia is extended to for the indication of Buddhist nun according to Shandong on January 29th, 2015,
In breakthrough treatment, which obtains extension approval, this is the huge ball egg of Waldenstrom that the first unique an example is ratified through FDA
White mass formed by blood stasis treatment.Ratified to list by European Drug Administration (EMA) for Buddhist nun according to Shandong, for treating jacket cell leaching in October, 2014
Bar tumor (MCL) and chronic lymphocytic leukemia (CLL).
Replacing Buddhist nun according to Shandong is bruton's tyrosine kinase (BTK) micromolecular inhibitor, residual with the cysteine of BTK active site
Base forms covalent bond, leads to BTK inhibition of enzyme activity.BTK is the letter of B- cell antigen receptor (BCR) and cytokine receptor access
Number molecule.The effect of BTK be by B- cell surface receptor issue signal instruction, thus activate B- cell transport, chemotactic, and
Stick required access.Non-clinical study, which is shown, inhibits malignant B-cells proliferation and existence in vivo according to Shandong for Buddhist nun, while also inhibiting body
Outer cell migration and substrate stick.Recurrent B- cell lymphoma patient takes according to Shandong for Buddhist nun, dosage >=2.5mg/kg/day (with
70 kilograms of weight meters per capita, daily dose, which is taken, per capita is greater than 175mg/day according to Shandong for Buddhist nun), peripheral blood is observed after 24 hours
Monocyte BTK active site is more than 90% to be captured.
The chemical name of Buddhist nun is replaced according to Shandong are as follows: 1- [(3R) -3- [4- amino -3- (4- Phenoxyphenyl) -1H pyrazolo [3,4-
D] pyrimidine -1- base] -1- piperidyl] -2- propylene -1- ketone, structural formula are as follows:
The prior art discloses seven kinds of process impurities or degradation impurity, they are obtained during synthesis replaces Buddhist nun according to Shandong,
However FDA examination & approval report only reported the impurity structure of chiral isomer and the structure of cylinder metabolism-ure (PCI-45227), to it
The structure of its impurity is not described.
It refers to according to Shandong in only document (patent WO2013/184572, the 90-91 pages) for the inspection of Buddhist nun A crystal form purity
The method of survey, this method are buffer (A) to 0.1% trifluoroacetic acid aqueous solution, are organic with the acetonitrile of 0.1% trifluoroacetic acid
Solvent (B), chromatographic column are Gemini-NX C18,4.6 × 150mm, 3 μm, and the HPLC method of gradient elution, this method is not
Specific impurities in preparation process are separated.
It is reported in Chinese Patent Application No. 201510165869.6 according to Shandong for the feather weight synthetic route of Buddhist nun, the route
Strategy are as follows: to be commercialized, the 3- of process route maturation (4- Phenoxyphenyl) -1H- pyrazolo [3,4-d] pyrimidine -4- amine
(SM1) and (S) -1- tertbutyloxycarbonyl -3- hydroxy piperidine (SM2) is starting material, different in triphenyl phosphorus and azoformic acid two
Under the action of propyl ester, Mitsunobu reaction is carried out in tetrahydrofuran solution system, reaction terminates, directly in the work of concentrated hydrochloric acid
With lower deprotection, then through obtaining intermediate -1:(R in sodium hydroxide and crystallization) -3- (4- Phenoxyphenyl) -1- (piperidines -3-
Base) -1H- pyrazolo [3,4-d] pyrimidine -4- amine.The intermediate is made in dichloromethane solvent with n,N-diisopropylethylamine
Alkali is condensed with acrylic anhydride and carries out amidation process, replaces Buddhist nun according to Shandong by ethanol/water system recrystallization finished product.
Above-mentioned process route has the advantages that the total recovery of (1) two-step reaction higher than other reports compared with other routes
The yield in road.(2) intermediate -1 and for the method for purification of Buddhist nun be recrystallization according to Shandong, makes to become according to Shandong for the industrialized production of Buddhist nun
It may；(3) triphenyl phosphorus for replacing polymer connection using cheap triphenyl phosphorus in Mitsunobu reaction, reduces work
Industry production cost；(4) " one kettle way " is used in the production of intermediate -1, Mitsunobu reaction is closed with deprotection reaction
And simplify technological operation.(5) amidation reagent is changed to acrylic anhydride by acryloyl chloride, avoids and is difficult to pass through recrystallization
The impurity of removal generates.The route is the synthetic strategy with industrial production value.Currently, there is no in this production line
The report of the process impurity of generation.
Summary of the invention
Impurity research is that the important content of drug research and development always through drug research and development directly affects the safety of drug
Property, validity and quality controllability.In order to provide related substance reference substance to according to Shandong for the quality research of Buddhist nun, raising is replaced according to Shandong
The quality standard of Buddhist nun, to provide important guidance according to Shandong for the safe medication of Buddhist nun, the present invention is by replacing Buddhist nun's technical study to according to Shandong
Process sampling carries out HPLC and LC-MS analysis, studies, synthesizes and confirms according to Shandong for the process impurity in Buddhist nun's production line.
What Chinese Patent Application No. 201510165869.6 was reported commercially available replaces for the synthetic method of Buddhist nun and according to Shandong according to Shandong
In Buddhist nun's capsule (Imbruvica is purchased from Pharmacyclics Janssen company, 140mg, lot number: L0404951), one can be related to
The process impurity of series, confirms that its chemical name and structural formula see the table below through inventor
The way of production of the above impurity is as follows:
Impurity A: this impurity is impurity in commercially available capsule, it should grind process impurity for original, in this technique is multiple batches of from
It is not detected.
Impurity B: in the synthetic method second step acyl for replacing Buddhist nun according to Shandong that Chinese Patent Application No. 201510165869.6 is reported
In aminating reaction, amidation occurs in two positions for replacing Buddhist nun's intermediate -1 (being expressed as YLTN-1) according to Shandong, piperidines nuclear nitrogen
And the nitrogen on pyrimidine ring, bisamide impurity in products B-1 is generated, due to Thermodynamic effect intramolecular cyclization occurs for the latter, raw
At more stable Sulfadiazine Compound ring impurity B.
Impurity C: in the synthetic method second step acyl for replacing Buddhist nun according to Shandong that Chinese Patent Application No. 201510165869.6 is reported
In aminating reaction, a small amount of acetic anhydride impurity is contained in starting material acrylic anhydride, it is miscellaneous that amidation process generation occurs with YLTN-1
Matter C.
Impurity D: in the synthetic method second step acyl for replacing Buddhist nun according to Shandong that Chinese Patent Application No. 201510165869.6 is reported
In aminating reaction, according to Shandong for Buddhist nun (YLTN) and by-product acrylic acid under the catalysis of alkali (may be n,N-diisopropylethylamine,
May be the nitrogen-containing heterocycle according to Shandong for Buddhist nun itself) Baylis-Hillman reaction occurs and generates impurity D.
Impurity E, F, G are according in the forced degradation sample of pharmaceutical preparation of the Shandong for Buddhist nun's active constituent or comprising replacing Buddhist nun according to Shandong
Principal degradation impurity, wherein impurity E is the primary product according to Shandong for Buddhist nun or according to Shandong for oxidative degradation in Buddhist nun's formulation process, miscellaneous
Matter F, G is for Buddhist nun or according to Shandong according to Shandong for the catabolite in Buddhist nun's formulation process, and replaces according to Shandong and force under Buddhist nun's alkaline condition
The main degradation products of degradation or hydrolysis.
Impurity H: in the synthetic method second step acyl for replacing Buddhist nun according to Shandong that Chinese Patent Application No. 201510165869.6 is reported
In aminating reaction, starting material acrylic anhydride contains a small amount of propionic andydride impurity, can react with intermediate -1 and generate impurity H:
As a result, in one aspect, the present invention relates to following compounds:
Compound A, chemical name are as follows: (R) -1- (3- (4- amino -3- (4- Phenoxyphenyl) -1H- pyrazolo [3,4-d]
Pyrimidine -1- base] piperidin-1-yl) -3- chloropropyl -1- ketone, and have a structure that
Compound B, chemical name are as follows: (R) -8- (1- acryloylpiperidine -3- base) -10- (4- Phenoxyphenyl) -3,4-
Dihydro-pyrazolo [4,3-e] pyrimido [1,2-c] pyrimidine -2 (8H) -one, and have a structure that
Compound C, chemical name are as follows: (R) -1-3- (4- amino -3- (4- Phenoxyphenyl) -1H- pyrazolo [3,4-d]
Pyrimidine -1- base] piperidin-1-yl)-ethyl ketone, and have a structure that
Compound D, chemical name are as follows: (R) -4- (3- (4- amino -3- (4- Phenoxyphenyl) -1H- pyrazolo [3,4-d]
Pyrimidine -1- base)-piperidines -1- carbonyl) amylene -4- acid, and have a structure that
Compound E, chemical name are as follows: 1- ((R) -1- acryloylpiperidine -3- base) -4- amino -3- (4- phenoxy group benzene
Base) -1H- pyrazolo [3,4-d] pyrimidine -1- oxide, and have a structure that
Compound F, chemical name are as follows: 1,3- bis- ((R) -3- (4- amino -3- (4- Phenoxyphenyl) -1H- pyrazoles [3,4-
D] pyrimidine -1- base) piperidin-1-yl) propane -1- ketone, and have a structure that
Compound G, chemical name are as follows: 1- ((R) -3- (4- ((3- ((R) -3- (4- amino -3- (4- Phenoxyphenyl) -
1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) -3- oxopropyl) ammonia) -3- (4- Phenoxyphenyl) -1H- pyrazoles [3,
4-d] pyrimidine -1- base) piperidin-1-yl) propyl -2- alkene -1- ketone, and have a structure that
Compound H, chemical name are as follows: (R) -1- (3- (4- amino -3- (4- Phenoxyphenyl) -1H- pyrazolo [3,4-d]
Pyrimidine -1- base)-piperidin-1-yl)-propyl -1- ketone, and have a structure that
In specific embodiments, above compound A-H is unpack format, preferably substantially pure form, more preferably
With greater than about 95% purity.
On the other hand, the present invention relates to the one or more of above compound A-H for Buddhist nun or to contain according to Shandong in detection according to Shandong
For the application in the sample purity of the pharmaceutical preparation of Buddhist nun as reference standard or reference substance, wherein being for the chemical name of Buddhist nun according to Shandong
1- [(3R) -3- [4- amino -3- (4- Phenoxyphenyl) -1H pyrazolo [3,4-d] pyrimidine -1- base] -1- piperidyl] -2- third
Alkene -1- ketone, structural formula are as follows:
Detailed description of the invention
Fig. 1: intermediate -1:(R) -3- (4- Phenoxyphenyl) -1- (piperidines -3- base) -1H- pyrazolo [3,4-d] pyrimidine -
4- amine1H-NMR；
Fig. 2: intermediate -1:(R) -3- (4- Phenoxyphenyl) -1- (piperidines -3- base) -1H- pyrazolo [3,4-d] pyrimidine -
The high resolution mass spectrum of 4- amine；
Fig. 3: Buddhist nun: 1- [(3R) -3- [4- amino -3- (4- Phenoxyphenyl) -1H pyrazolo [3,4-d] pyrimidine-is replaced according to Shandong
1- yl] -1- piperidyl] -2- propylene -1- ketone1H-NMR；
Fig. 4: Buddhist nun: 1- [(3R) -3- [4- amino -3- (4- Phenoxyphenyl) -1H pyrazolo [3,4-d] pyrimidine-is replaced according to Shandong
1- yl] -1- piperidyl] -2- propylene -1- ketone high resolution mass spectrum.
Fig. 5: impurity A: (R) -1- (3- (4- amino -3- (4- Phenoxyphenyl) -1H- pyrazolo [3,4-d] pyrimidine -1-
Base] piperidin-1-yl) -3- chloropropyl -1- ketone1H-NMR；
Fig. 6: impurity A: (R) -1- (3- (4- amino -3- (4- Phenoxyphenyl) -1H- pyrazolo [3,4-d] pyrimidine -1-
Base] piperidin-1-yl) -3- chloropropyl -1- ketone high resolution mass spectrum；
Fig. 7: impurity B: (R) -8- (1- acryloylpiperidine -3- base) -10- (4- Phenoxyphenyl) -3,4- pyrazoline
And [4,3-e] pyrimido [1,2-c] pyrimidine -2 (8H) -one1H-NMR；
Fig. 8: impurity B: (R) -8- (1- acryloylpiperidine -3- base) -10- (4- Phenoxyphenyl) -3,4- pyrazoline
And the high resolution mass spectrum of [4,3-e] pyrimido [1,2-c] pyrimidine -2 (8H) -one；
Fig. 9: impurity C:(R) -1-3- (4- amino -3- (4- Phenoxyphenyl) -1H- pyrazolo [3,4-d] pyrimidine -1- base]
Piperidin-1-yl)-ethyl ketone1H-NMR；
Figure 10: impurity C:(R) -1-3- (4- amino -3- (4- Phenoxyphenyl) -1H- pyrazolo [3,4-d] pyrimidine -1-
Base] piperidin-1-yl)-ethyl ketone high resolution mass spectrum.
Figure 11: impurity D:(R) -4- (3- (4- amino -3- (4- Phenoxyphenyl) -1H- pyrazolo [3,4-d] pyrimidine -1-
Base)-piperidines -1- carbonyl) amylene -4- acid1H-NMR；
Figure 12: impurity D:(R) -4- (3- (4- amino -3- (4- Phenoxyphenyl) -1H- pyrazolo [3,4-d] pyrimidine -1-
Base)-piperidines -1- carbonyl) amylene -4- acid high resolution mass spectrum；
Figure 13: impurity H:((R) -1- (3- (4- amino -3- (4- Phenoxyphenyl) -1H- pyrazolo [3,4-d] pyrimidine -1-
Base)-piperidin-1-yl)-propyl -1- ketone1H-NMR；
Figure 14: impurity H:(R) -1- (3- (4- amino -3- (4- Phenoxyphenyl) -1H- pyrazolo [3,4-d] pyrimidine -1-
Base)-piperidin-1-yl)-propyl -1- ketone high resolution mass spectrum；
Figure 15: impurity E: 1- ((R) -1- acryloylpiperidine -3- base) -4- amino -3- (4- Phenoxyphenyl) -1H- pyrrole
Azoles simultaneously [3,4-d] pyrimidine -1- oxide1H-NMR。
Figure 16: impurity E: 1- ((R) -1- acryloylpiperidine -3- base) -4- amino -3- (4- Phenoxyphenyl) -1H- pyrrole
The high resolution mass spectrum of azoles simultaneously [3,4-d] pyrimidine -1- oxide.
Figure 17: impurity F: ((R) -3- (4- amino -3- (4- Phenoxyphenyl) -1H- pyrazoles [3,4-d] pyrimidine-of 1,3- bis-
1- yl) piperidin-1-yl) propane -1- ketone1H-NMR。
Figure 18: impurity F: ((R) -3- (4- amino -3- (4- Phenoxyphenyl) -1H- pyrazoles [3,4-d] pyrimidine-of 1,3- bis-
1- yl) piperidin-1-yl) propane -1- ketone high resolution mass spectrum.
Figure 19: impurity G:1- ((R) -3- (4- ((3- ((R) -3- (4- amino -3- (4- Phenoxyphenyl) -1H- pyrazoles [3,
4-d] pyrimidine -1- base) piperidin-1-yl) -3- oxopropyl) ammonia) -3- (4- Phenoxyphenyl) -1H- pyrazoles [3,4-d] pyrimidine -
1- yl) piperidin-1-yl) propyl -2- alkene -1- ketone1H-NMR。
Figure 20: impurity G:1- ((R) -3- (4- ((3- ((R) -3- (4- amino -3- (4- Phenoxyphenyl) -1H- pyrazoles [3,
4-d] pyrimidine -1- base) piperidin-1-yl) -3- oxopropyl) ammonia) -3- (4- Phenoxyphenyl) -1H- pyrazoles [3,4-d] pyrimidine -
1- yl) piperidin-1-yl) propyl -2- alkene -1- ketone high resolution mass spectrum；
Specific embodiment
In order to which further the present invention will be described, will be illustrated below by specific embodiment, but reality below
Example is applied to be not limited in any way protection scope of the present invention.
Embodiment 1: intermediate -1:(R) -3- (4- Phenoxyphenyl) -1- (piperidines -3- base) -1H- pyrazolo [3,4-d]
The synthesis of pyrimidine -4- amine (YLTN-1)
155mL tetrahydrofuran is added into the reaction flask of 500mL, 3- (4- Phenoxyphenyl) -1H- is sequentially added under stirring
Pyrazoles [3,4-D] pyrimidine -4- amine (SM1) (5g, 1eq), (S) -1- tertbutyloxycarbonyl -3- hydroxy piperidine (SM2) (4.97g,
1.5eq), triphenylphosphine (13g, 3.0eq).Under the conditions of 25 DEG C of temperature control, in 30 minutes, diisopropyl azodiformate is added dropwise
Tetrahydrofuran solution (by 10g, 3.0eq diisopropyl azodiformate is dissolved in the tetrahydrofuran of 10mL).After being added dropwise to complete, control
25 DEG C of temperature, the reaction was continued 5 hours (TLC monitoring: ethyl acetate: methanol=10:1).The lower vacuum distillation of stirring.15 DEG C of temperature control, to
30mL concentrated hydrochloric acid is added dropwise in residue, time for adding 30 minutes, after being added dropwise to complete, 25 DEG C of temperature control, the reaction was continued 2 hours.With two
Chloromethanes aqueous phase extracted three times (each 75mL), retains water phase.It is primary with the extraction of 50mL ethyl acetate.Remaining water phase is sufficiently stirred
It mixes, 15 DEG C of temperature control, the sodium hydrate aqueous solution of about 45g 20% is added dropwise, monitor reaction solution using pH test paper, pH=5~6 (are added dropwise
About 30 minutes time) obtain light yellow solid.Twice by ethyl alcohol recrystallization by above-mentioned solid, off-white powder 2.87g is obtained, received
Rate: 45%.1H-NMR(400Mz,DMSO-d6) δ: 8.226 (s, 1H), 7.656~7.634 (m, 2H), 7.435~7.395 (m,
2H), 7.185~7.094 (m, 5H), 4.689~4.635 (m, 1H), 3.081~3.043 (m, 1H), 2.957~2.930 (m,
1H), 2.901~2.873 (m, 1H), 2.490~2.457 (m, 1H), 2.125~2.015 (m, 2H), 1.750~1.717 (m,
1H), 1.589~1.518 (m, 1H) (being detailed in Fig. 1)；ESI-HRMS spectrogram shows molecular ion peak m/z=387.19449 [MW+
H]+, corresponding molecular weight is consistent with the structural formula calculated value (387.19279) of offer.Absolute error is 4.41ppm,
Within high resolution mass spectrum error range.(being detailed in Fig. 2)
Embodiment 2: the preparation of Buddhist nun is replaced according to Shandong
Under nitrogen protection, 50mL methylene chloride, intermediate -1 (5g, 1eq), N, N- are sequentially added into 100mL there-necked flask
Diisopropylethylamine (2g, 1.2eq).Under the conditions of -10 DEG C of temperature control, start the dichloromethane that acrylic anhydride (1.96g, 1.2eq) is added dropwise
Alkane solution, time for adding 30 minutes, after being added dropwise to complete, solution became clarification by muddiness, keeps the temperature -10 DEG C, it is anti-to be stirred well to raw material
It should completely (TLC detection, methanol: ethyl acetate: triethylamine=1:5:0.05).Reaction solution 5% aqueous citric acid solution of 200mL
Washing, removes water phase, and methylene chloride is evaporated off in concentration.Get Yi Lu replaces Buddhist nun's crude product: 3.63g.1H-NMR(400Mz,DMSO-d6)δ:
8.259 (s, 1H), 7.654~7.674 (m, 2H), 7.405~7.444 (m, 2H), 7.109~7.195 (m, 5H), 6.676~
6.743 (m, 1H), 6.046~6.152 (m, 1H), 5.570~5.713 (m, 1H), 4.690~4.716 (m, 1H), 4.554~
4.583 (m, 0.5H), 4.208 (m, 1H), 4.052~4.085 (m, 0.5H), 3.674~3.731 (m, 0.5H), 3.184~
3.214 (m, 1H), 2.972~3.027 (m, 0.5H), 2.245~2.282 (m, 1H), 2.128 (m, 1H), 1.903~1.937
(m, 1H), 1.577~1.605 (m, 1H) (being detailed in Fig. 3)；ESI-HRMS spectrogram shows molecular ion peak: 441.20366 [MW+H]+,
The calculated value of Buddhist nun's molecular ion peak is replaced according to Shandong are as follows: 441.20335 [M+H]+, absolute error 0.71ppm meets high-resolution
Mass spectrum error range, measured value are consistent with theoretical value.(being detailed in Fig. 4).
Embodiment 3: impurity A: (R) -1- (3- (4- amino -3- (4- Phenoxyphenyl) -1H- pyrazolo [3,4-d] pyrimidine -
1- yl] piperidin-1-yl) -3- chloropropyl -1- ketone synthesis.
Under nitrogen protection, 50ml methylene chloride is added into the there-necked flask of 100mL, sequentially adds intermediate -1 under stirring:
(R) -3- (4- Phenoxyphenyl) -1- (piperidines -3- base) -1H- pyrazoles [3,4-d] pyrimidine -4- amine (YLTN-1) (1.00g,
1eq), n,N-diisopropylethylamine (0.40g, 1.2eq) is cooled to -20~-10 DEG C, start be added dropwise 3- chlorpromazine chloride (1.46g,
1eq), it is added dropwise, solution becomes clarification by muddiness, continues stirring 20~30 minutes, and LC-MS detection, raw material disappears, and decompression is steamed
It evaporates, methylene chloride to no fraction is evaporated off and steams, obtains crude product and is purified with column chromatography method, elute ratio are as follows: methanol: ethyl acetate
=1:10, collects the total 400mL of eluent, and vacuum distillation is evaporated off solvent to no fraction and steams.Obtaining off-white powder 830mg is
(R) -1- (3- (4- amino -3- (4- Phenoxyphenyl) -1H- pyrazolo [3,4-d] pyrimidine -1- base] piperidin-1-yl) -3- chlorine third
Base -1- ketone.1H-NMR(400Mz,CDCl3) δ: 8.382 (d, J=13.2Hz, 1H), 7.660~7.638 (m, 2H), 7.419~
7.380 (m, 2H), 7.200~7.083 (m, 5H), 5.751 (m, 2H), 4.891~4.853 (m, 1H), 4.842~4.812 (m,
0.5H), 4.558 (m, 0.5H), 4.066~4.033 (m, 0.5H), 3.844~3.833 (m, 1H), 3.773~3.746 (m,
0.5H), 3.355 (m, 0.5H), 3.191 (m, 0.5H), 2.877~2.843 (m, 2H), 2.812~2.795 (m, 1H), 2.434
~2.270 (m, 2H), 2.048~1.955 (m, 1H), 1.756~1.689 (m, 1H).(being detailed in Fig. 5) ESI-HRMS spectrogram is shown
Molecular ion peak m/z=477.17930 [MW+H]+, the structural formula calculated value of corresponding molecular weight and offer
(476.17275) it is consistent.Absolute error is 1.52ppm, within high resolution mass spectrum error range.(being detailed in Fig. 6).Impurity A exists
Replacing the relative retention time (RRT) of Buddhist nun according to Shandong with respect to principal component on HPLC spectrogram is about 1.06.
Embodiment 4: impurity B: (R) -8- (1- acryloylpiperidine -3- base) -10- (4- Phenoxyphenyl) -3,4- dihydro
The synthesis of pyrazolo [4,3-e] pyrimido [1,2-c] pyrimidine -2 (8H) -one.
100ml methylene chloride is added into the there-necked flask of 250mL, sequentially adds intermediate -1 under stirring for nitrogen protection:
(R) -3- (4- Phenoxyphenyl) -1- (piperidines -3- base) -1H- pyrazoles [3,4-d] pyrimidine -4- amine (YLTN-1) (2.00g,
1eq), n,N-diisopropylethylamine (1.30g, 2.0eq) is cooled to -20 DEG C~-10 DEG C.Start be added dropwise acrylic anhydride (1.33g,
2eq), -10 DEG C of process temperature control or less is added dropwise；Drop finishes, and continues stirring 20~30 minutes, and LC-MS detection has target product generation,
30~40 DEG C of temperature control, vacuum degree: -0.08MPa, vacuum distillation are evaporated off methylene chloride to no fraction and steam；Obtain crude product column layer
Methods For Purification is analysed, elutes ratio are as follows: methanol: ethyl acetate=1:5, collection eluent, 30~40 DEG C of temperature control, vacuum degree :-
0.08MPa, vacuum distillation, is evaporated off solvent to no fraction and steams.Obtaining off-white powder 531mg is (R) -8- (1- acryloyl group piperazine
Pyridine -3- base) -10- (4- Phenoxyphenyl) -3,4- dihydro-pyrazolo [4,3-e] pyrimido [1,2-c] pyrimidine -2 (8H) -one.1H-NMR(400Mz,CDCl3) δ: 8.352 (m, 2H), 7.874 (m, 2H), 7.367~7.328 (m, 2H), 7.144~7.050
(m, 5H), 6.591~6.558 (m, 1H), 6.335~6.293 (m, 1H), 5.747~5.658 (m, 1H), 4.803 (m, 1H),
4.649~4.622 (m, 0.5H), 4.300 (m, 0.5H), 4.333~4.300 (m, 2H), 4.153 (m, 0.5H), 4.051~
4.019 (m, 0.5H), 3.751~3.680 (m, 0.5H), 3.354~3.217 (m, 1H), 2.902 (m, 0.5H), 2.789~
2.770 (m, 2H), 2.397~2.223 (m, 2H), 2.194~2.022 (m, 1H), 2.005~1.718 (m, 1H) (are detailed in figure
7).ESI-HRMS spectrogram shows molecular ion peak m/z=495.21580 [MW+H]+, the structure of corresponding molecular weight and offer
Formula calculated value (494.20664) is consistent.Absolute error is 3.82ppm, (is detailed in figure within high resolution mass spectrum error range
8).It is about 0.97 that impurity B, which replaces the relative retention time (RRT) of Buddhist nun according to Shandong with respect to principal component on HPLC spectrogram,.
Embodiment 5: impurity C:(R) -1-3- (4- amino -3- (4- Phenoxyphenyl) -1H- pyrazolo [3,4-d] pyrimidine -
1- yl] piperidin-1-yl)-ethyl ketone synthesis
By intermediate -1:(R) -3- (4- Phenoxyphenyl) -1- (piperidines -3- base) -1H- pyrazoles [3,4-d] pyrimidine -4- amine
(YLTN-1) (2.00g, 1eq) is dissolved in methylene chloride (50ml), addition N, N- diisopropyl ethyl amine (0.80g, 2eq),
- 10 DEG C are cooled to, nitrogen protection is added dropwise acetic anhydride (0.53g, 1eq), finishes, and reacts 30 minutes, and LC-MS monitoring has been reacted
At system being concentrated to dryness, residue is by silica gel column chromatography, and eluent: ethyl acetate: it is white to obtain class for methanol=10:1
Color powder 1.38g be (R) -1-3- (4- amino -3- (4- Phenoxyphenyl) -1H- pyrazolo [3,4-d] pyrimidine -1- base] piperidines -
1- yl)-ethyl ketone.1H-NMR(400Mz,CDCl3) δ: 8.352 (d, J=16Hz, 1H), 7.657~7.623 (m, 2H), 7.402~
7.364 (m, 2H), 7.187~7.069 (m, 5H), 5.971 (m, 2H), 4.849~4.841 (m, 0.5H), 4.841~4.821
(m, 1H), 4.574~4.541 (m, 0.5H), 4.045~4.005 (m, 0.5H), 3.864~3.831 (m, 0.5H), 3.744~
3.685 (m, 0.5H), 3.318~3.253 (m, 0.5H), 3.201~3.141 (m, 0.5H), 2.802~2.745 (m, 0.5H),
2.403~2.249 (m, 2H), 1.962~1.697 (m, 2H) (being detailed in Fig. 9)；ESI-HRMS spectrogram shows molecular ion peak m/z
=429.20416 [MW+H]+, corresponding molecular weight is consistent with the structural formula calculated value (428.19607) of offer.Absolutely
Error is 1.89ppm, (is detailed in Figure 10) within high resolution mass spectrum error range.Impurity C opposite principal component on HPLC spectrogram
The relative retention time (RRT) that Buddhist nun is replaced according to Shandong is about 0.96.
Embodiment 6: impurity D:(R) -4- (3- (4- amino -3- (4- Phenoxyphenyl) -1H- pyrazolo [3,4-d] pyrimidine -
1- yl)-piperidines -1- carbonyl) amylene -4- acid synthesis
Under nitrogen protection, Buddhist nun 1- [(3R) -3- [4- amino -3- (4- Phenoxyphenyl) -1H pyrazolo [3,4- will be replaced according to Shandong
D] pyrimidine -1- base] -1- piperidyl] -2- propylene -1- ketone (2g, 1eq) is dissolved in methylene chloride (50ml), sequentially add N, N-
Diisopropyl ethyl amine (0.70g, 2eq), acrylic acid (0.98g, 3eq), reaction are warming up to 35 DEG C, stir 36 hours, by system
It is concentrated to dryness, residue is by silica gel column chromatography, and eluent: ethyl acetate: methanol=8:1 obtains off-white powder 103mg
For (R) -4- (3- (4- amino -3- (4- Phenoxyphenyl) -1H- pyrazolo [3,4-d] pyrimidine -1- base)-piperidines -1- carbonyl) penta
Alkene -4- acid,1H-NMR(400Mz,DMSO-d6) δ: 12.309 (br, 1H), 8.263 (S, 1H), 7.687~7.666 (m, 2H),
7.422~7.461 (m, 2H), 7.122~7.211 (m, 5H), 6.009~6.072 (m, 0.5H), 5.570~5.660 (m,
0.5H), 4.647~4.772 (m, 1H), 4.516~4.547 (m, 0.5H), 4.200-4.232 (m, 0.5H), 4.069-4.101
(m, 0.5H), 3.913~3.945 (m, 0.5H), 3.629-3.655 (m, 0.5H), 3.129~3.157 (m, 1H), 2.878 (m,
0.5H), 2.445-2.513 (m, 4H), 2.206~2.264 (m, 1H), 2.085~2.125 (m, 1H), 1.861~1.936 (m,
1H), 1.522~1.666 (m, 1H) (being detailed in Figure 11)；ESI-HRMS spectrogram shows molecular ion peak m/z=513.22606 [MW+
H]+, corresponding molecular weight is consistent with the structural formula calculated value (512.21720) of offer.Absolute error is 3.09ppm,
(Figure 12 is detailed in) within high resolution mass spectrum error range.Impurity D is opposite for Buddhist nun according to Shandong with respect to principal component on HPLC spectrogram
Retention time (RRT) is about 1.02.
Embodiment 7: impurity H:(R) -1- (3- (4- amino -3- (4- Phenoxyphenyl) -1H- pyrazolo [3,4-d] pyrimidine -
1- yl)-piperidin-1-yl)-propyl -1- ketone synthesis
Under nitrogen protection, by intermediate -1:(R) -3- (4- Phenoxyphenyl) -1- (piperidines -3- base) -1H- pyrazoles [3,4-
D] pyrimidine -4- amine (YLTN-1) (2.00g, 1eq) dissolution methylene chloride (50mL) in, be added N, N- diisopropyl ethyl amine
(0.80g, 2eq) is cooled to -10 DEG C, and nitrogen protection is added dropwise propionic andydride (0.67g, 1eq), finishes, and reacts 30 minutes, LC-MS
Monitoring, reaction are completed, system are concentrated to dryness, and residue passes through silica gel column chromatography, eluent: ethyl acetate: methanol=
10:1, obtaining lightpink powder 1.72g is that ((4- amino -3- (4- Phenoxyphenyl) -1H- pyrazolo [3,4-d] is phonetic by 3- by (R) -1-
Pyridine -1- base)-piperidin-1-yl)-propyl -1- ketone.1H-NMR(400Mz,CDCl3) δ: 8.338~8.302 (m, 2H), 7.658~
7.627 (m, 2H), 7.440~7.400 (m, 2H), 7.224~7.097 (m, 5H), 4.881~4.634 (m, 1H), 4.881~
4.826 (m, 1H), 4.108~3.094 (m, 1H), 2.762 (m, 1H), 3.310~3.167 (m, 1H), 2.452~2.315 (m,
4H), 1.213~1.157 (m, 3H), 2.069~1.691 (m, 2H) (being detailed in Figure 13)；ESI-HRMS spectrogram shows molecular ion
Peak m/z=443.25380 [MW+H]+, corresponding molecular weight and structural formula calculated value (442.21172) phase of offer
Symbol.Absolute error is 3.66ppm, (is detailed in Figure 14) within high resolution mass spectrum error range.Impurity H is opposite on HPLC spectrogram
Principal component is about 1.04 for the relative retention time (RRT) of Buddhist nun according to Shandong.
Embodiment 8: impurity E: 1- ((R) -1- acryloylpiperidine -3- base) -4- amino -3- (4- Phenoxyphenyl) -1H-
The preparation of pyrazolo [3,4-d] pyrimidine -1- oxide
(1) preparation of impurity E
Prepared by Example 2 replaces Buddhist nun about 5.17g according to Shandong, sets in 500ml reaction flask, adds 80% acetonitrile-water of 150ml molten
Solution, then plus 30% hydrogen peroxide 30ml, be stirred overnight at room temperature, reaction terminates, and sodium thiosulfate is added into reaction solution, until bubble-free
Generate, 35 DEG C be concentrated under reduced pressure into no liquid outflow, then plus 3 times of water phase volumes ethyl acetate, extraction, collect organic phase, merge,
35 DEG C are concentrated to dryness, and get Yi Lu replaces Buddhist nun's impurity E crude product 2.65g.It is 51.26% that LC-MS, which detects crude product purity, liquid chromatogram
On relative to according to Shandong for Ni Feng relative retention time be 0.98, mass-to-charge ratio [MW+H]+It is 457.2.Impurity E1H-NMR figure ginseng
See Figure 15, high resolution mass spectrum is referring to Figure 16.
(2) purification of impurity E crude product
The impurity E crude product of the above-mentioned preparation of 2.95g is dissolved into the solution that concentration is about 150mg/ml with DMSO, use is following
Preparative chromatography is further purified:
Chromatographic column (purchased from Ai Jieer science and technology): using 10 μm of partial size of C18 filler, column (50mm is filled after being homogenized with isopropanol
×250mm)；
Mobile phase: water and acetonitrile
Flow velocity: 120ml/min；
Detection wavelength: 254nm；
Sample volume: 10ml
Gradient elution, elution program are as follows:
| Time (min) | Mobile phase A (%) | Mobile phase B (%) |
| 0 | 65 | 35 |
| 10 | 65 | 35 |
| 30 | 45 | 55 |
| 50 | 45 | 55 |
| 50.01 | 20 | 80 |
| 55 | 20 | 80 |
| 55.01 | 65 | 35 |
| 60 | 65 | 35 |
Collect fraction corresponding to the peak that retention time is 30~37min；The purity of LC-MS or HPLC checked for impurities E.
35 DEG C are concentrated under reduced pressure collected fraction to no liquid and flow out, and the ethyl acetate extraction of 3 times of volumes is added, and collection has
Machine phase merges, anhydrous sodium sulfate is added to dry, filter, and 30 DEG C of filtrate are concentrated to dryness, and obtains 2.06g light pink solid, passes through
HPLC measurement, purity 98.97%, total recovery 36.6%.
(3) Structural Identification of impurity E
The high resolution mass spectrum (source ESI) of impurity E shows its mass-to-charge ratio [M+H]+It is 457.19930, corresponding molecular weight
Absolute error with the structural formula calculated value 456.19099 of offer is 2.27ppm, meets the error model of high resolution mass spectrum
It encloses.The structure for replacing Buddhist nun according to Shandong with reference to formula (2), in conjunction with impurity E1H-NMR、13C-NMR spectrum identifies its structural formula as shown in following formula E.
Impurity E high resolution mass spectrum shows that its molecular weight is consistent completely with provided molecular formula；
The nuclear magnetic resonance of 1 impurity E of table1H-NMR、13C-NMR parsing result
The shown information of the nuclear magnetic resonance spectroscopy of impurity E, carbon spectrum can be to provided formula E molecular structure of compounds formula
Upper hydrogen, carbon all belong to.Therefore, it is consistent for Buddhist nun's impurity E with provided formula E molecular structural formula according to Shandong.
Embodiment 9: impurity F: ((4- amino -3- (4- Phenoxyphenyl) -1H- pyrazoles [3,4-d] is phonetic by (R) -3- by 1,3- bis-
Pyridine -1- base) piperidin-1-yl) propane -1- ketone and impurity G:1- ((R) -3- (4- ((3- ((R) -3- (4- amino -3- (4- phenoxy group
Phenyl) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) -3- oxopropyl) ammonia) -3- (4- Phenoxyphenyl) -1H-
Pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl -2- alkene -1- ketone preparation.
(1) preparation of impurity F and impurity G
The preparation of about 20g embodiment 2 is added in 1000ml reaction flask replaces Buddhist nun according to Shandong, and 250ml acetonitrile and 200ml is added
5M sodium hydrate aqueous solution, 80 DEG C of heating in water bath for reaction 40min, is cooled to room temperature, and 5M HCl is added to be neutralized to neutrality, 35 DEG C of decompressions
It is concentrated into no liquid to reserve, the ethyl acetate extraction of 5 times of water phase volumes is added, collects upper organic phase, merges, 35 DEG C of decompressions are dense
It is reduced to dry, obtains according to Shandong for Buddhist nun's impurity F, G crude product 18.72g, LC-MS detects crude product purity, according to corresponding MS quasi-molecular ions, liquid phase
With respect to for the peak that the relative retention time of Ni Feng is about 0.94 (impurity F) and 1.15 (impurity G) being object according to Shandong in chromatography,
Content in crude product is respectively 16.9% (impurity F) and 18.4% (impurity G).Impurity F1H-NMR figure is referring to Figure 17, high-resolution
Mass spectrum is referring to Figure 18.Impurity G's1H-NMR figure is referring to Figure 19, high resolution mass spectrum referring to fig. 20.
(2) purification of impurity F and impurity G crude product
DMSO is added to be dissolved into the solution of 10ml, loading the crude product of the 18.72g impurity F of above-mentioned preparation, G；
It is middle to suppress standby column (being purchased from Agela company): Agela XBP C18 filler, 480g, packing material size: 20~35 μm
Mobile phase: water (0.05%TFA) and pure acetonitrile
Flow velocity: 40ml/min
Detection wavelength: 254nm
Applied sample amount :~5g
Gradient elution, elution program are as follows:
| Time (min) | Mobile phase A (%) | Mobile phase B (%) |
| 0 | 75 | 25 |
| 5 | 75 | 25 |
| 30 | 65 | 35 |
| 40 | 65 | 35 |
| 50 | 50 | 50 |
| 55 | 50 | 50 |
| 55.01 | 75 | 25 |
| 60 | 75 | 25 |
The fraction at all peaks is collected respectively, and LC-MS or HPLC confirm target product.Collect the fraction (impurity of 30~33min
F) and the fraction (impurity G) of 40~44min, merge identical fraction, be concentrated under reduced pressure respectively, until no liquid flows out, be added 3 in water phase
The ethyl acetate extraction of times volume, collects organic phase, anhydrous sodium sulfate is added to dry, filter, collect filtrate.It is concentrated to dryness,
Get Yi Lu replaces Buddhist nun's impurity G secondary separation crude product 2.57g for Buddhist nun's impurity F sterling 2.21g and Yi Lu respectively.It is examined through LC-MS or HPLC
Analysis is surveyed, it is 98.15% that the purity of Buddhist nun's impurity F is replaced according to Shandong, and it is 85.32% that the purity of Buddhist nun's impurity G secondary separation crude product is replaced according to Shandong.
(3) secondary separation of impurity G
The 2.57g of above-mentioned preparation adds 10ml DMSO to dissolve according to Shandong for Buddhist nun's impurity G secondary separation crude product
According to Shandong for condition used by the G secondary separation of Buddhist nun's impurity are as follows:
Chromatographic column: Waters XBridge C8 30mm × 100mm, 5 μm
Mobile phase: water and acetonitrile
Detection wavelength: 254nm
Flow velocity: 25ml/min
Gradient elution, elution program are as follows:
Sampling volume: 2ml/ times every time；
The fraction of retention time 26min~30min is collected, and confirms object purity with LC-MS.Merge identical fraction,
35 DEG C are concentrated under reduced pressure into the outflow of no acetonitrile, add the ethyl acetate of 3 times of volumes extract, merge organic phase, add anhydrous sodium sulfate drying,
Filtering, is concentrated to dryness, and obtains replacing Buddhist nun's impurity G sterling 1.64g according to Shandong, white solid powder, and LC-MS detection purity is
95.13%.
(4) Structural Identification of impurity F
The high resolution mass spectrum (source ESI) of impurity F shows its mass-to-charge ratio [M+H]+It is 827.39065, corresponding molecular weight
Absolute error with the structural formula calculated value 826.34520 of offer is 2.16ppm, meets the error model of high resolution mass spectrum
It encloses.The high resolution mass spectrum of sample shows that its molecular weight is consistent completely with provided molecular formula.With reference to the knot for replacing Buddhist nun according to Shandong
Structure, in conjunction with this product1H-NMR、13C-NMR, H-HCOSY, HMBC, HSQC, DEPT spectrum identify its structural formula as shown in following formula F.
Table 2 is impurity F1H-NMR, H-HCOSY and HMBC parse data
3 impurity F of table13The parsing of C-NMR, DEPT, HSQC, HMBC data
By sample1H-NMR, H-HCOSY and HMBC spectrum it is found that in its structure Hydrogen Proton number and formula F compound knot
Structure is consistent.By sample13C-NMR, DEPT, HSQC and HMBC compose the structure it is found that carbon number and formula F compound in its structure
It is consistent.Nucleus magnetic hydrogen spectrum, carbon spectrum and two-dimentional modal data can rationally be belonged to and are associated with carbon to the hydrogen on structural formula.
(5) Structural Identification of impurity G
The high resolution mass spectrum (source ESI) of impurity G shows its mass-to-charge ratio [M+H]+It is 881.40140, corresponding molecular weight
Absolute error with the structural formula calculated value 880.39215 of offer is 1.43ppm, meets the error model of high resolution mass spectrum
It encloses.The high resolution mass spectrum of sample shows that its molecular weight is consistent completely with provided molecular formula.In conjunction with the knot for replacing Buddhist nun according to Shandong
Structure, in conjunction with this product1H-NMR、13C-NMR, DEPT, H-HCOSY, HMBC and hsqc spectrum identify its structure as shown in following formula G.
Table 4 is impurity G's1H-NMR, H-HCOSY and HMBC data and hydrogen ownership
Table 5 is impurity G's13C-NMR, DEPT, HSQC and HMBC data and carbon ownership
According to sample1H-NMR spectrum, in conjunction with H-HCOSY, HMBC two-dimensional spectrum it is found that Hydrogen Proton number and formula G in its structure
The structure of compound is consistent.By sample13C-NMR, DEPT, HSQC and HMBC compose carbon number and formula Gization it is found that in its structure
The structure for closing object is consistent.
Embodiment 10: impurity A-H is replacing the application in Buddhist nun's defects inspecting as reference substance according to Shandong
(1) preparation of impurity A-H contrast solution:
Take the impurity A-H of above-mentioned preparation appropriate, it is accurately weighed, it is set in 100ml volumetric flask respectively, adds appropriate acetonitrile (wherein
Impurity F and G add 2~4ml DMSO) ultrasound makes to dissolve, then plus acetonitrile be settled to scale, shake up, it is impure that every 1ml be respectively prepared
The solution of each about 10 μ g of A-H, then it is accurate measure solution 1.0ml, set in 10ml volumetric flask, 80% acetonitrile-water added to be diluted to quarter
Degree, shake up, both respectively containing according to Shandong for Buddhist nun's impurity A-H reference substance solution.
(2) preparation of test solution:
The preparation that Buddhist nun's crude product test solution is replaced according to Shandong of above-mentioned preparation: take it is prepared in the above embodiments according to Shandong for Buddhist nun's sample
Crude product about 10mg, it is accurately weighed, it sets in 10ml volumetric flask, adds 80% acetonitrile-water to dissolve and be settled to scale, shake up, be made every
1ml contains the solution that Buddhist nun about 1mg is replaced according to Shandong, replaces Buddhist nun's bulk pharmaceutical chemicals test solution as according to Shandong.
The preparation of Buddhist nun's capsule test solution is replaced according to Shandong: taking that (Imbruvica is purchased from according to Shandong for Buddhist nun's capsule
Pharmacyclics Janssen company, 140mg, lot number: L0404951) content fine powder it is appropriate (containing according to Shandong for Buddhist nun about
10mg), accurately weighed, it sets in 10ml volumetric flask, adds 80% acetonitrile-water to dissolve and be settled to scale, shake up, filter, discard just
Filtrate 2ml takes subsequent filtrate to be used as according to Shandong for Buddhist nun's capsule test solution.
(3) chromatographic condition is as follows:
With octyl silane group silica gel C8 column (4.6 × 250mm, 5 μm) as chromatographic column (purchased from Ai Jieer science and technology), flowing
Phase are as follows: water (0.05%TFA) and acetonitrile, flow velocity 1.2ml/min, Detection wavelength 254nm, 35 DEG C of column temperature, gradient elution, elution
Program is as follows:
| Time (min) | Mobile phase A (%) | Mobile phase B (%) |
| 0 | 80 | 20 |
| 3 | 80 | 20 |
| 15 | 20 | 80 |
| 22 | 20 | 80 |
| 22.01 | 80 | 20 |
| 30 | 80 | 20 |
(4) test procedure: taking 10 μ l of above-mentioned impurity contrast solution and test solution, is injected separately into liquid chromatograph (purchase
From Agilent company of the U.S., model 1260) or LC-MS instrument (Agilent company of the U.S., Agilent 6120), record color
Spectrogram, it is qualitative for the relative retention time progress of Buddhist nun according to Shandong to principal component according to each impurity phase, according to external standard method (Chinese Pharmacopoeia
Version in 2010, two, annex V D) calculate the content of impurity A-H.
(4) test result:
1. commercially available replace impurity spectrum analysis in Buddhist nun's capsule according to Shandong:
| Serial number | Content (%) | [MW+H]+ | Impurity ownership | Relative retention time |
| The impurity 1 detected | 0.01 | 429.2 | C | 0.96 |
| The impurity 2 detected | 0.05 | 495.2 | B | 0.97 |
| The impurity 3 detected | 0.05 | 443.2 | H | 1.04 |
| The impurity 4 detected | 0.12 | 477.2 | A | 1.06 |
| The impurity 5 detected | 0.13 | 414.2/827.4 | F | 0.94 |
| The impurity 6 detected | 0.04 | 441.3/881.3 | Unknown impuritie | 1.03 |
| The impurity 7 detected | 0.08 | 441.3/881.6 | G | 1.15 |
2. the impurity spectrum analysis for replacing Buddhist nun's crude product according to Shandong of above-mentioned preparation
| Serial number | Content (%) | [MW+H]+ | Impurity ownership | Relative retention time |
| The impurity 1 detected | 0.01 | 429.2 | C | 0.96 |
| The impurity 2 detected | 0.41 | 495.2 | B | 0.97 |
| The impurity 3 detected | 0.70 | 443.2 | H | 1.04 |
| The impurity 4 detected | 0.06 | 513.2 | D | 1.02 |
| The impurity 5 detected | 0.14 | 414.2/827.4 | F | 0.94 |
| The impurity 6 detected | 0.07 | 441.3/881.3 | Unknown impuritie | 1.03 |
| The impurity 7 detected | 0.03 | 441.3/881.6 | G | 1.15 |
| The impurity 8 detected | 0.07 | 457.2 | E | 0.98 |
It should be noted that all documents referred in the present invention are incorporated as referring in this application, just as each
Piece document is individually recited as with reference to such.In addition, it should also be understood that, above-described is that specific implementation of the invention is arranged and transported
Technical principle, after having read the contents of the present invention, those skilled in the art can do various changes or be repaired to the present invention
Change without departing from spirit and scope of the invention, such equivalent forms are also fallen in the scope of the present invention.
Claims (5)
1. compound B, chemical name are as follows: (R) -8- (1- acryloylpiperidine -3- base) -10- (4- Phenoxyphenyl) -3,4- bis-
Hydrogen pyrazolo [4,3-e] pyrimido [1,2-c] pyrimidine -2 (8H) -one, and have a structure that
2. compound D, chemical name are as follows: ((4- amino -3- (4- Phenoxyphenyl) -1H- pyrazolo [3,4-d] is phonetic by 3- by (R) -4-
Pyridine -1- base)-piperidines -1- carbonyl) amylene -4- acid, and have a structure that
3. compound E, chemical name are as follows: 1- ((R) -1- acryloylpiperidine -3- base) -4- amino -3- (4- Phenoxyphenyl) -
1H- pyrazolo [3,4-d] pyrimidine -1- oxide, and have a structure that
4. compound H, chemical name are as follows: ((4- amino -3- (4- Phenoxyphenyl) -1H- pyrazolo [3,4-d] is phonetic by 3- by (R) -1-
Pyridine -1- base)-piperidin-1-yl)-propyl -1- ketone, and have a structure that
5. compound A, compound B, compound C, compound D, compound E, compound F, one kind of compound G and compound H
It is a variety of detection according to Shandong for Buddhist nun or containing according to Shandong for application as reference substance in the sample purity of the pharmaceutical preparation of Buddhist nun, wherein
The chemical name that Buddhist nun is replaced according to Shandong is 1- [(3R) -3- [4- amino -3- (4- Phenoxyphenyl) -1H pyrazolo [3,4-d] pyrimidine -1-
Base] -1- piperidyl] -2- propylene -1- ketone, structural formula are as follows:
Wherein
Compound A title are as follows: (R) -1- (3- (4- amino -3- (4- Phenoxyphenyl) -1H- pyrazolo [3,4-d] pyrimidine -1-
Base] piperidin-1-yl) -3- chloropropyl -1- ketone, and have a structure that
Compound B title are as follows: (R) -8- (1- acryloylpiperidine -3- base) -10- (4- Phenoxyphenyl) -3,4- pyrazoline
And [4,3-e] pyrimido [1,2-c] pyrimidine -2 (8H) -one, and have a structure that
Compound C title are as follows: (R) -1-3- (4- amino -3- (4- Phenoxyphenyl) -1H- pyrazolo [3,4-d] pyrimidine -1- base]
Piperidin-1-yl)-ethyl ketone, and have a structure that
Compound D title are as follows: (R) -4- (3- (4- amino -3- (4- Phenoxyphenyl) -1H- pyrazolo [3,4-d] pyrimidine -1-
Base)-piperidines -1- carbonyl) amylene -4- acid, and have a structure that
Compound E title are as follows: 1- ((R) -1- acryloylpiperidine -3- base) -4- amino -3- (4- Phenoxyphenyl) -1H- pyrazoles
And [3,4-d] pyrimidine -1- oxide, and have a structure that
Compound F title are as follows: ((R) -3- (4- amino -3- (4- Phenoxyphenyl) -1H- pyrazoles [3,4-d] pyrimidine -1- of 1,3- bis-
Base) piperidin-1-yl) propane -1- ketone, and have a structure that
Compound G title are as follows: 1- ((R) -3- (4- ((3- ((R) -3- (4- amino -3- (4- Phenoxyphenyl) -1H- pyrazoles [3,
4-d] pyrimidine -1- base) piperidin-1-yl) -3- oxopropyl) ammonia) -3- (4- Phenoxyphenyl) -1H- pyrazoles [3,4-d] pyrimidine -
1- yl) piperidin-1-yl) propyl -2- alkene -1- ketone, and have a structure that
Compound H title are as follows: (R) -1- (3- (4- amino -3- (4- Phenoxyphenyl) -1H- pyrazolo [3,4-d] pyrimidine -1-
Base)-piperidin-1-yl)-propyl -1- ketone, and have a structure that
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510188801.XA CN106146516B (en) | 2015-04-20 | 2015-04-20 | The preparation method of Buddhist nun's impurity of the drug is replaced according to Shandong |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510188801.XA CN106146516B (en) | 2015-04-20 | 2015-04-20 | The preparation method of Buddhist nun's impurity of the drug is replaced according to Shandong |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN106146516A CN106146516A (en) | 2016-11-23 |
| CN106146516B true CN106146516B (en) | 2019-02-15 |
Family
ID=58057659
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201510188801.XA Active CN106146516B (en) | 2015-04-20 | 2015-04-20 | The preparation method of Buddhist nun's impurity of the drug is replaced according to Shandong |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN106146516B (en) |
Families Citing this family (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107014943B (en) * | 2017-06-07 | 2018-08-21 | 福建省微生物研究所 | A kind of detection method for replacing Buddhist nun's enantiomter according to Shandong |
| CN108440540A (en) * | 2018-05-16 | 2018-08-24 | 杭州中美华东制药有限公司 | A kind of her cloth is for Buddhist nun's compound and preparation method thereof |
| CN108623602B (en) * | 2018-05-31 | 2020-07-17 | 厦门医学院 | Method for preparing and purifying ibrutinib |
| CN115716832A (en) * | 2021-08-25 | 2023-02-28 | 深圳鼎邦化学品有限公司 | Preparation method and application of ibrutinib impurity |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102656173A (en) * | 2009-10-12 | 2012-09-05 | 药品循环公司 | Inhibitors of Bruton's tyrosine kinase |
| CN104407067A (en) * | 2014-11-17 | 2015-03-11 | 广东东阳光药业有限公司 | Ibrutinib and test method of isomer of ibrutinib |
-
2015
- 2015-04-20 CN CN201510188801.XA patent/CN106146516B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102656173A (en) * | 2009-10-12 | 2012-09-05 | 药品循环公司 | Inhibitors of Bruton's tyrosine kinase |
| CN104407067A (en) * | 2014-11-17 | 2015-03-11 | 广东东阳光药业有限公司 | Ibrutinib and test method of isomer of ibrutinib |
Also Published As
| Publication number | Publication date |
|---|---|
| CN106146516A (en) | 2016-11-23 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN106146516B (en) | The preparation method of Buddhist nun's impurity of the drug is replaced according to Shandong | |
| Cao et al. | Design, synthesis and 3D-QSAR of β-carboline derivatives as potent antitumor agents | |
| CN106153798B (en) | It is a kind of to be used to analyze the purposes for doing reference standard about the HPLC methods and these impurity of material for Buddhist nun's preparation according to Shandong for Buddhist nun and according to Shandong | |
| KR20070088485A (en) | Purification of cinacalcet | |
| CN108017620A (en) | The methanesulfonic acid uncommon preparation method for Buddhist nun's process contaminants difficult to understand | |
| CN104086445B (en) | A kind of bromhexine hydrochloride and preparation have the detection method of related substance | |
| CN103123342B (en) | The impurity analysis preparation method of clindamycin | |
| CN103804357B (en) | A kind of Rupatadine fumarate compound, its synthetic method and pharmaceutical composition thereof | |
| CN111961079A (en) | Ruideciclovir related substance and preparation method and application thereof | |
| Barandun et al. | From lin‐Benzoguanines to lin‐Benzohypoxanthines as Ligands for Zymomonas mobilis tRNA–Guanine Transglycosylase: Replacement of Protein–Ligand Hydrogen Bonding by Importing Water Clusters | |
| CN106153797B (en) | It is a kind of to replace Buddhist nun's preparation Related substance method according to Shandong for Buddhist nun and according to Shandong | |
| AU2011328598B2 (en) | Radiolabelled phenylimidazole-based ligands | |
| CN106146512B (en) | The preparation method of Buddhist nun is replaced according to Shandong | |
| CN104098557B (en) | The preparation of a kind of Rupatadine fumarate impurity J and detection method | |
| CN103864646A (en) | Preparation and analysis method of impurity of rasagiline mesylate | |
| CN112110897A (en) | Preparation method of deuterated crizotinib and derivative thereof | |
| CN110885331B (en) | Preparation and application of 6-amino-1H-pyrazolo [3, 4-d ] pyrimidine JAK kinase inhibitor | |
| CN113004244A (en) | Trelagliptin impurity and preparation method and application thereof | |
| CN110452248A (en) | A kind of novelty sesquiterpenoid and the preparation method and application thereof | |
| CN106619636A (en) | Delafloxacin impurity compounds and preparation methods thereof | |
| CN111440168A (en) | Preparation method and application of 6-methoxy paliperidone palmitate | |
| CN106478506B (en) | The preparation method of half light green Ka Selin hydrochloride | |
| CN109912568A (en) | A kind of acetal compound, preparation method and the usage | |
| CN106153772B (en) | A method of with the related substance of the miscellaneous Shandong amine of high performance liquid chromatography detection grace | |
| BUCHI REDDY et al. | Identification and characterization of potential impurities in raloxifene hydrochloride |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |