CN114276341A - LS007 impurity compound A and preparation process and application thereof - Google Patents
LS007 impurity compound A and preparation process and application thereof Download PDFInfo
- Publication number
- CN114276341A CN114276341A CN202110755792.3A CN202110755792A CN114276341A CN 114276341 A CN114276341 A CN 114276341A CN 202110755792 A CN202110755792 A CN 202110755792A CN 114276341 A CN114276341 A CN 114276341A
- Authority
- CN
- China
- Prior art keywords
- substance
- reaction
- compound
- preparation
- stirring
- 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.)
- Pending
Links
Images
Landscapes
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention belongs to the technical field of chemical pharmacy, and particularly relates to an LS007 impurity compound A and a preparation process and application thereof. The preparation process of the LS007 impurity compound A comprises the following steps: taking a substance A1-5, hydrochloric acid and a sodium nitrite solution as main raw materials to carry out a sixth reaction; wherein the structural formula of the substance A1-5 is shown in the specification
Description
Technical Field
The invention belongs to the technical field of chemical pharmacy, and particularly relates to an LS007 impurity compound A, and a preparation process, a raw material formula and application thereof.
Background
Proliferative diseases such as cancer are characterized by uncontrolled and unregulated cell proliferation. Protein kinases are an important class of enzymes for cancer research. The protein kinase family is one of the largest families in the human genome, and most kinases have a conserved functional domain consisting of 300 amino acid residues (250-300) constituting a catalytic domain, which contains a binding site for ATP, the phosphate group of which is covalently transferred to a substrate molecule to phosphorylate the substrate. Protein kinases can be classified according to the type of substrate they act on, for example, serine kinases, threonine kinases, tyrosine kinases, and the like.
Kinases are enzymes that catalyze the transfer of phosphate groups from high energy molecules containing phosphate to specific substrates, mediate intracellular signaling pathway activation, a variety of extracellular and other stimuli, act as molecular switches that can modulate or regulate the biological functions of target proteins, and trigger kinases to phosphorylate substrates. Extracellular stimuli can affect one or more cellular responses related to cell growth, migration, differentiation, hormone secretion, transcription factor activation, muscle contraction, glucose metabolism, protein synthesis control, and cell cycle regulation, among others. If the kinase mediates an abnormal cellular response, it will lead to a variety of diseases including, but not limited to, allergy and asthma, alzheimer's disease, autoimmune diseases, bone diseases, cancer, cardiovascular diseases, inflammatory diseases, hormone-related diseases, metabolic diseases, neurological diseases, neurodegenerative diseases, and the like.
Numerous molecules capable of inhibiting the function of protein kinases by blocking ATP binding are known in the prior art. Cyclin-dependent kinases (CDKs) are serine/threonine protein kinases associated with various cyclin subunits, playing a key role in regulating cell cycle processes and transcription cycles. 10 different CDKs (CDK1-9 and 11) are involved in a variety of important regulatory pathways in eukaryotic cells, including cell cycle control, apoptosis, neuronal physiology, differentiation and transcription. ). In addition, CDKs, particularly CDK2, CDK7, and CDK9, are also required for viral replication processes. Inhibitors of CDKs that limit viral replication have been reported to act on a number of viruses, including human immunodeficiency virus, human cytomegalovirus, herpes virus and varicella-zoster virus.
Among them, inhibitors of CDK9 are a new strategy for the current potential treatment of cardiovascular diseases, including cardiac hypertrophy. Cardiac hypertrophy is characterized by an increase in the total amount of mRNA and protein synthesis, and CDK7 and CDK9 are the major drivers of transcription and are closely associated with cardiac hypertrophy. Thus, inhibition of CDK9 and its associated cyclins is an effective therapeutic strategy for cardiovascular diseases.
Inhibitors of CDKs may also be useful in the treatment of neurodegenerative disorders such as alzheimer's disease. The appearance of double-stranded Helical Filaments (Pair Helical fibers) associated with Alzheimer's disease was caused by the hyperphosphorylation of Tau protein by CDK5/p 25.
Chinese patent publication No. CN103373994A (incorporated herein by reference in its entirety) discloses a compound with CDK-9 inhibitory function and a preparation method thereof; the Chinese patent with publication number CN108658966A discloses tartrate and a crystal form of a compound 3- (5-fluoro-4- (4-methyl-2- (methylamino) thiazol-5-yl) pyrimidin-2-ylamino) -benzenesulfonamide (hereinafter, referred to as a compound LS007), which has good performance and potential of being developed into a selective CDK9 kinase inhibitor.
However, currently, research on impurities and quality control thereof in the synthesis process of the compound 3- (5-fluoro-4- (4-methyl-2- (methylamino) thiazol-5-yl) pyrimidin-2-ylamino) -benzenesulfonamide is still blank.
Disclosure of Invention
The invention provides an LS007 impurity compound A and a preparation process and application thereof.
In order to solve the technical problem, the invention provides a preparation process of a compound (namely LS007 impurity compound A), which comprises the following steps: taking a substance A1-5, hydrochloric acid and a sodium nitrite solution as main raw materials to carry out a sixth reaction; wherein the structural formula of the substance A1-5 is shown in the specification
In a second aspect, the present invention provides a compound having the formula
In a third aspect, the present invention provides the use of a compound as hereinbefore described as a standard for the analysis of a pharmaceutical compound.
The invention has the beneficial effects that the compound and the synthesis method thereof are provided for the first time, the compound is one of main impurities in the synthesis process of LS007, and the compound has important significance for the quality control of LS 007. The compound of the invention is prepared by a sixth reaction by taking the substance A1-5, hydrochloric acid and sodium nitrite solution as main raw materials, and can be used as a standard substance for medicinal compound analysis.
Additional features and advantages of the invention will be set forth in the description which follows, and in part will be obvious from the description, or may be learned by practice of the invention. The objectives and other advantages of the invention will be realized and attained by the structure particularly pointed out in the written description and claims hereof as well as the appended drawings.
In order to make the aforementioned and other objects, features and advantages of the present invention comprehensible, preferred embodiments accompanied with figures are described in detail below.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below, and it is obvious that the drawings in the following description are some embodiments of the present invention, and other drawings can be obtained by those skilled in the art without creative efforts.
FIG. 1 is a process flow diagram for the preparation of compounds of the present invention;
FIG. 2 is a flow chart of a process for the preparation of substance A1-5 according to the invention;
FIG. 3 is a flow chart of a process for the preparation of substance A1-4 according to the invention;
FIG. 4 is a flow chart of a process for the preparation of substance A1-3 according to the invention;
FIG. 5 is a flow chart of a process for the preparation of substance A1-2 according to the invention;
FIG. 6 is a flow chart of a process for the preparation of substance A1-1 according to the invention;
FIG. 7 is a graph of an infrared spectrum of a compound of the present invention;
FIG. 8 is a NMR spectrum of a compound of the invention;
FIG. 9 is a NMR carbon spectrum of a compound of the invention;
FIG. 10 is a mass spectrum of a compound of the present invention;
FIG. 11 is a system suitability HPLC chromatogram obtained by adding LS007 to the impurity compound A and compound A1-3 synthesized by the present invention as standard reference substances;
FIG. 12 is an HPLC chromatogram of LS007 synthesized according to the prior art.
Detailed Description
To make the objects, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions of the present invention will be clearly and completely described below with reference to the accompanying drawings, and it is apparent that the described embodiments are some, but not all embodiments of the present invention. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
The first part
Disadvantages of the prior art to this end, and with reference to fig. 1, the present invention provides a process for the preparation of a compound comprising: taking a substance A1-5, hydrochloric acid and a sodium nitrite solution as main raw materials to carry out a sixth reaction; wherein the structural formula of the substance A1-5 is shown in the specification
Alternatively, referring to fig. 1, the sixth reaction comprises: dropwise adding a substance A1-5 and hydrochloric acid into a sodium nitrite solution in an ice bath, and reacting at room temperature; slowly pouring the reaction solution into water to separate out solid; stirring and filtering the solid; and drying to obtain the compound. Specifically, adding a substance A1-5 and hydrochloric acid into a flask, dropwise adding a sodium nitrite solution in an ice bath, reacting for 1h at room temperature after dropwise adding, slowly pouring a reaction solution into water, precipitating yellow solids, stirring for 1h, filtering, and drying to obtain the compound.
The reaction formula of the sixth reaction is
As an alternative to the preparation of substance A1-5.
Referring to fig. 2, the preparation method of the substance a1-5 comprises the following steps: taking a substance A1-3, a substance A1-4 and ethylene glycol monomethyl ether as main raw materials to carry out a fifth reaction; wherein the structural formula of the substance A1-3 is shown in the specification
The structural formula of the substance A1-4 is shown in the specification
The fifth reaction comprises: rough preparation and refining; wherein the crude comprises: mixing and stirring the substance A1-3, the substance A1-4 and ethylene glycol monomethyl ether; heating to 124-126 ℃, and reacting under a reflux system; cooling the reaction solution to below 60 ℃, and transferring the reaction solution to ice water; stirring at room temperature; performing suction filtration to obtain a filter cake; leaching the filter cake with water to obtain a crude product; the refining comprises: adding the crude product into ethylene glycol monomethyl ether, and stirring at 25 +/-5 ℃; performing suction filtration to obtain a filter cake; leaching the filter cake with ethylene glycol monomethyl ether; drying at 50 + -5 deg.C to obtain substance A1-5; the reaction formula of the fifth reaction is:
specifically, crude: adding a substance A1-3 and a substance A1-4 into a reaction kettle, adding ethylene glycol monomethyl ether, stirring, heating, and reacting at 124-126 ℃ (system reflux) for 22 +/-2 hours; cooling to reduce the temperature of the reaction mixed solution to below 60 ℃, then slowly transferring the feed liquid into 22L ice water, and stirring at room temperature for about 0.5 hour; and (4) carrying out suction filtration, and leaching a filter cake with a small amount of water to obtain a crude product. Refining: and (3) putting the crude product into a four-mouth bottle, adding ethylene glycol monomethyl ether, stirring for 2 hours at the temperature of 25 +/-5 ℃, carrying out suction filtration, leaching a filter cake by using a proper amount of ethylene glycol monomethyl ether, and drying the obtained filter cake by hot air (50 +/-5 ℃) to obtain a brown solid, namely the substance A1-5. Optionally, in the fifth reaction, the molar ratio of the substance A1-3 to the substance A1-4 is 1: 1.05-1.2, preferably 1: 1.1.
as an alternative to the preparation of substance A1-4.
Referring to fig. 3, the preparation method of the substance a1-4 comprises: performing a fourth reaction by using m-aminobenzenesulfonamide, cyanamide and acetonitrile as main raw materials; the fourth reaction comprises: mixing and stirring m-aminobenzenesulfonamide, cyanamide and acetonitrile; cooling to 5 +/-1 ℃ and dropwise adding trimethylchlorosilane; heating to reflux, and keeping the temperature for reaction; cooling the reaction mixture to room temperature; performing suction filtration to obtain a filter cake; leaching the filter cake with acetonitrile; and drying at 60 +/-2 ℃ to obtain the substance A1-4; the reaction formula of the fourth reaction is
Concretely, m-aminophenylsulfonamide, cyanamide and acetonitrile are added into a reaction bottle and stirred; cooling to 5 +/-1 ℃, dropwise adding trimethyl chlorosilane, after dropwise adding, heating to reflux, and carrying out heat preservation reaction for 30 +/-1 hours; cooling the reaction mixture to room temperature, carrying out suction filtration to obtain a filter cake, leaching with a small amount of acetonitrile, and drying in hot air at 60 +/-2 ℃ to obtain the substance A1-4. Optionally, in the fourth reaction, the molar ratio of the m-aminobenzenesulfonamide to the cyanamide to the trimethylchlorosilane to the substance A1-3 to the substance A1-4 is 1: 1.05-1.2: 1.05-1.2, preferably 1: 1.1: 1.1.
as an alternative to the preparation of substance A1-3.
Referring to fig. 4, the preparation method of the substance a1-3 comprises: carrying out a third reaction by taking a substance A1-2, methanol, dichloromethane, acetonitrile and a fluorine reagent as main raw materials; wherein the structural formula of the substance A1-2 is shown in the specification
The third reaction comprises: mixing and stirring the substance A1-2, methanol, dichloromethane and acetonitrile; cooling to-5 +/-1 ℃, and adding a fluorine reagent selectfluor for reaction; adding ammonia methanol solution (the concentration is 8mol/L) to quench the reaction, and continuing stirring; rotary steaming at 30 ℃; adding water, stirring and filtering; air-drying at room temperature to obtain crystalline solid; and column chromatography purification to obtain the substance A1-3; the reaction formula of the third reaction is
Specifically, the substance A1-2, methanol, dichloromethane and acetonitrile are added into a reaction kettle, stirred, cooled to minus 5 +/-1 ℃, added with a fluorine reagent, reacted for 0.5 hour, added with an ammonia methanol solution, continuously stirred overnight, subjected to rotary evaporation at 30 ℃, added with water, stirred for 30 minutes, filtered, air-dried at room temperature to obtain a yellow crystalline solid, and subjected to column chromatography purification to obtain the substance A1-3. Optionally, in the third reaction, the molar ratio of the A1-2 to the fluorine reagent and the ammonia methanol is 1: 1-1.5: 1-5, preferably 1: 1-1.5: 1-3; further, 1: 1: 2.
as an alternative to the preparation of substance A1-2.
Referring to fig. 5, the preparation method of the substance a1-2 comprises: carrying out a second reaction by taking the substance A1-1, N-dimethylformamide dimethyl acetal and xylene as main raw materials; wherein the structural formula of the substance A1-1 is shown in the specification
The second reaction comprises: mixing and stirring the substance A1-1, N-dimethylformamide dimethyl acetal and xylene; heating to reflux, and keeping the temperature for reaction; evaporating the solution under reduced pressure to obtain viscous solid; cooling to 60 ℃, and adding methanol into the viscous solid; heating to reflux; cooling and crystallizing; filtering to obtain a filter cake; leaching the filter cake with methanol; drying at 60 +/-2 ℃ to obtain a substance A1-2; the second reaction has the formula
Specifically, adding the substance A1-1, N-dimethylformamide dimethyl acetal and xylene into a reaction bottle, stirring, heating to reflux, and reacting for 24 +/-1 hours under heat preservation; and after the reaction is finished, evaporating the solution under reduced pressure to obtain an orange viscous solid, cooling to 60 ℃, adding methanol, heating to reflux for 1 hour, cooling, crystallizing, filtering, leaching a filter cake with a small amount of methanol, and drying in hot air at 60 +/-2 ℃ to obtain the substance A1-2. Alternatively, in the second reaction, the substance A1-1And the molar ratio of N, N-dimethylformamide dimethyl acetal is 1: (1.5-3), preferably 1: 1.9.
as an alternative to the preparation of substance A1-1.
Referring to fig. 6, the preparation method of the substance a1-1 comprises: carrying out a first reaction by taking N-methylthiourea, methanol, pyridine and 3-chloro-2, 4-pentanedione as main raw materials; wherein the first reaction comprises: mixing and stirring N-methylthiourea and methanol; adding pyridine to form a white suspension; cooling to 0-5 ℃, and dropwise adding 3-chloro-2, 4-pentanedione; preserving the heat at 30-35 ℃ and reacting; pouring the reaction solution into water, and continuously stirring; performing suction filtration to obtain a filter cake; leaching the filter cake with water; drying at 80 +/-2 ℃ to obtain a substance A1-1; the reaction formula of the first reaction is
Specifically, adding N-methylthiourea and methanol into a reaction bottle, stirring, adding pyridine, and presenting a white suspension; cooling to 0-5 ℃, and dropwise adding 3-chloro-2, 4-pentanedione; and after the dropwise addition is finished, keeping the temperature of 30-35 ℃ for reacting for 2 hours, pouring the reaction solution into water, continuously stirring for half an hour, carrying out suction filtration, leaching the filter cake for 3 times with water, and drying the filter cake with hot air at the temperature of 80 +/-2 ℃ to obtain a substance A1-1. Optionally, in the first reaction, the molar ratio of the N-methylthiourea to the pyridine to the 3-chloro-2, 4-pentanedione is 1: (1-1.2): (1 to 1.5), preferably 1: 1:1.
in the present application, the first reaction, the second reaction, the third reaction, the fourth reaction, the fifth reaction and the sixth reaction do not necessarily show a certain order relationship, but are merely reactions occurring when preparing each substance (substance A1-1, substance A1-2, substance A1-3, substance A1-4 and substance A1-5) for convenience of distinction. Meanwhile, the first reaction, the second reaction, the third reaction, the fourth reaction, the fifth reaction and the sixth reaction may be only reaction processes, and may also include processes in which reactions are performed, reactant addition or pretreatment processes before the reactions, and purification or cleaning processes of reaction products after the reactions, so that the variations in the above descriptions do not affect the variations in the reactions in the present application. In addition, the substance A1-1, the substance A1-2, the substance A1-3, the substance A1-4 and the substance A1-5 are used for distinguishing each intermediate product for preparing the compound, wherein A1-1, A1-2, A1-3, A1-4 and A1-5 are only the number of the substance and have no limit to the components and the structure of each substance.
Further, the invention provides a compound, and the structural formula of the compound is
Further, the present invention provides the use of a compound as described hereinbefore as a standard for the analysis of pharmaceutical compounds.
Alternatively, the pharmaceutical compound is for example, but not limited to LS007(CDK kinase inhibitor targeted anticancer drugs, pyrimidine derivative drugs targeting protein kinases for anticancer). The compounds may be used as impurities in the pharmaceutical compounds for use as standards in the analysis of pharmaceutical compounds.
Specifically, the specific method of using the compound as an impurity control (hereinafter, impurity a) is as follows:
taking a proper amount of the compound, precisely weighing, adding a proper amount of dimethyl sulfoxide to dissolve, and diluting with acetonitrile-water (1: 1) as a solvent to prepare a solution containing 0.4mg in 1mL as a test solution; taking a proper amount of the substance A1-3, the impurity A and LS007, adding a proper amount of solvent, and carrying out ultrasonic treatment to dissolve and dilute the substance A1-3, the impurity A and the LS007 to prepare mixed solutions containing about 4 mu g of each substance per 1mL as system applicability solutions. Measuring by high performance liquid chromatography (0512 in the four-department general regulation of 2020 edition in China pharmacopoeia).
As shown in fig. 11, 10 μ L of the system suitability solution was precisely measured and injected into a liquid chromatograph, and the chromatogram was recorded. From the figure, the relative retention times of LS007, impurity Compound A and substance A1-3 can be seen, thus enabling the use of the compounds as standards for the analysis of pharmaceutical compounds.
The second part
Example 1
(1) Preparation of substance A1-1
100.5g N-methylthiourea (molecular weight: 90.1, 1.12mol) and 477g of methanol were put into a 1L reaction flask, stirred, and 118.5g of pyridine (molecular weight: 79.1, 1.34mol) was added to the flask to give a white suspension. The temperature is reduced to 0-5 ℃, and 226g of 3-chloro-2, 4-pentanedione (molecular weight 134.6, 1.68mol) is added dropwise. After the dropwise addition, reacting for 2 hours at the temperature of 30-35 ℃, pouring the reaction solution into 3L of water, continuously stirring for half an hour, carrying out suction filtration, leaching the filter cake for 3 times, and drying the filter cake by hot air at the temperature of 80 +/-2 ℃ to obtain 166g of a substance A1-1 (with the molecular weight of 170.2 and the purity of 0.976mol), wherein the purity is 99% and the yield is 86.3%.
(2) Preparation of substance A1-2
161.8g of substance A1-1 (molecular weight: 170.2, 0.95mol), 333.8g N, N-dimethylformamide dimethyl acetal (molecular weight: 119.2, 2.8mol) and 220g of xylene were put into a 1L reaction flask, stirred, heated to reflux, and reacted for 24. + -. 1 hours while maintaining the temperature. After the reaction is finished, the solution is evaporated under reduced pressure to obtain an orange viscous solid, the orange viscous solid is cooled to 60 ℃, 220mL of methanol is added, then the orange viscous solid is heated to reflux for 1 hour, the temperature is reduced, crystallization is carried out, filtration is carried out, a filter cake is leached by a small amount of methanol, and the filter cake is dried in hot air at the temperature of 60 +/-2 ℃, so that 123.9g of a substance A1-2 (the molecular weight is 225.3, 0.55mol), the purity is 98 percent and the yield is 57 percent.
(3) Preparation of substance A1-3
56.8g of the substance A1-2 (molecular weight: 225.3, 0.25mol), 450mL of methanol, 680mL of dichloromethane and 450mL of acetonitrile were put into A3L reaction vessel, stirred, cooled to-5. + -. 1 ℃, added with 131.1g of a fluorine reagent (molecular weight: 354.3, 0.37mol), reacted for 0.5 hour, added with 63.3g of an ammonia methanol solution (ammonia concentration: 8mol/L), stirred overnight, evaporated at 30 ℃, added with 500g of water, stirred for 30 minutes, filtered, air-dried at room temperature to obtain a yellow crystalline solid, and purified by column chromatography to obtain 34.1g of the substance A1-3 (molecular weight: 243.3, 0.140mol), purity: 95% and yield: 53%.
(4) Preparation of substance A1-4
67.5g of m-aminobenzenesulfonamide (molecular weight: 172.2, 0.39mol), 19.7g of cyanamide (molecular weight: 42.0, 0.47mol) and 660mL of acetonitrile were put into a 1L reaction flask and stirred. Cooling to 5 +/-1 ℃, dropwise adding 51.0g of trimethylchlorosilane (molecular weight of 108.6, 0.47mol), after dropwise adding, heating to reflux, and carrying out heat preservation reaction for 30 +/-1 hours. The reaction mixture is cooled to room temperature, a filter cake is obtained by suction filtration, a small amount of acetonitrile is used for leaching, and the filter cake is dried in hot air at the temperature of 60 +/-2 ℃, so that 78.6g of the substance A1-4 (with the molecular weight of 250.7, 0.33mol) is obtained, the purity is 95 percent, and the yield is 80 percent.
(5) Preparation of substance A1-5
Crude preparation: adding 10g of substance A1-3 (with molecular weight of 243.3, 0.041mol) and 12.3g of substance A1-4 with molecular weight of 250.7, 0.049mol) into a reaction kettle, adding 100g of ethylene glycol monomethyl ether (with molecular weight of 76.1,1.31mol), stirring, heating, and reacting for 22 +/-2 hours at 124-126 ℃ (system reflux); cooling to reduce the temperature of the reaction mixed solution to below 60 ℃, then slowly transferring the feed liquid into 22L ice water, and stirring at room temperature for about 0.5 hour; and (4) carrying out suction filtration, and leaching a filter cake with a small amount of water to obtain a crude product.
Refining: and (3) putting the crude product into a four-mouth bottle, adding 18g of ethylene glycol monomethyl ether, stirring at 25 +/-5 ℃ for 2 hours, carrying out suction filtration, eluting a filter cake by using an appropriate amount of ethylene glycol monomethyl ether, and drying the obtained filter cake by hot air (50 +/-5 ℃) to obtain 12.1g of brown solid, namely a substance A1-5 (the molecular weight is 430.9, 0.028mol), the purity is 98 percent and the yield is 67 percent.
(6) Preparation of the Compounds
10g of substance A1-5 (molecular weight of 430.9, 0.023mol) and 0.2L of hydrochloric acid are added into a flask, 10g of aqueous solution of sodium nitrite (containing 4g of sodium nitrite and 0.058mol) is dropwise added under ice bath, reaction is carried out for 1h at room temperature, reaction liquid is slowly poured into 1L of water to precipitate yellow solid, the mixture is stirred for 1h, and the mixture is filtered and dried to obtain 4.2g of compound (molecular weight of 395.4, 0.011mol), the purity is 98 percent and the yield is 47.8 percent.
Example 2
(1) Preparation of substance A1-1
100.5g N-methylthiourea (molecular weight: 90.1, 1.12mol) and 477g of methanol were put into a 1L reaction flask, stirred, and 88.2g of pyridine (molecular weight: 79.1, 1.12mol) was added to the flask to give a white suspension. The temperature is reduced to 0-5 ℃, and 150g of 3-chloro-2, 4-pentanedione (molecular weight 134.6, 1.11mol) is added dropwise. After the dropwise addition, the reaction is carried out for 2 hours at the temperature of 30-35 ℃, the reaction solution is poured into 2L of water, the stirring is continued for half an hour, the suction filtration is carried out, the filter cake is leached by the water for 3 times, and the filter cake is dried by hot air at the temperature of 80 +/-2 ℃ to obtain 164g of substance A1-1 (the molecular weight is 170.2, 0.964mol), the purity is 99 percent, and the yield is 86.0 percent.
(2) Preparation of substance A1-2
161.8g of substance A1-1 (molecular weight: 170.2, 0.95mol), 214.9g N, N-dimethylformamide dimethyl acetal (molecular weight: 119.2, 1.8mol) and 176.3g of xylene were put into a 1L reaction flask, stirred, heated to reflux, and reacted for 24. + -. 1 hours while maintaining the temperature. After the reaction is finished, the solution is evaporated under reduced pressure to obtain an orange viscous solid, the orange viscous solid is cooled to 60 ℃, 200mL of methanol is added, then the orange viscous solid is heated to reflux for 1 hour, the temperature is reduced, crystallization is carried out, filtration is carried out, a filter cake is leached by a small amount of methanol, and the filter cake is dried in hot air at the temperature of 60 +/-2 ℃, so that 114.7g of a substance A1-2 (the molecular weight is 225.3, 0.51mol) is obtained, the purity is 98%, and the yield is 52%.
(3) Preparation of substance A1-3
56.8g of substance A1-2 (molecular weight: 225.3, 0.25mol), 430mL of methanol, 648mL of methylene chloride and 430mL of acetonitrile are put into A3L reaction kettle, stirred, cooled to-5 + -1 ℃, 89.4g of fluorine reagent (molecular weight: 354.3, 0.25mol) is added, reaction is carried out for 0.5 hour, 63.3g of ammonia methanol solution (ammonia concentration: 8mol/L) is added, stirring is continued overnight, the solvent is evaporated at 30 ℃, 500g of water is added, stirring is carried out for 30 minutes, then filtration is carried out, air drying is carried out at room temperature, yellow crystalline solid is obtained, and column chromatography purification is carried out, so as to obtain 32.1g of substance A1-3 (molecular weight: 243.3, 0.132mol), purity is 95%, and yield is 50%.
(4) Preparation of substance A1-4
67.5g of m-aminobenzenesulfonamide (molecular weight: 172.2, 0.39mol), 18.1g of cyanamide (molecular weight: 42.0, 0.43mol) and 600mL of acetonitrile were put into a 1L reaction flask and stirred. Cooling to 5 +/-1 ℃, dropwise adding 46.7g of trimethylchlorosilane (molecular weight of 108.6, 0.43mol), after dropwise adding, heating to reflux, and carrying out heat preservation reaction for 30 +/-1 hours. The reaction mixture is cooled to room temperature, a filter cake is obtained by suction filtration, a small amount of acetonitrile is used for leaching, and the filter cake is dried in hot air at the temperature of 60 +/-2 ℃, so that 77g of the substance A1-4 (with the molecular weight of 250.7 and the molecular weight of 0.31mol) is obtained, the purity is 95 percent, and the yield is 75 percent.
(5) Preparation of substance A1-5
Crude preparation: adding 10g of substance A1-3 (with molecular weight of 243.3, 0.041mol) and 11.3g of substance A1-4 with molecular weight of 250.7, 0.045mol) into a reaction kettle, adding 93.4g of ethylene glycol monomethyl ether (with molecular weight of 76.1,1.23mol), stirring, heating, and reacting at 124-126 ℃ (system reflux) for 22 +/-2 hours; cooling to reduce the temperature of the reaction mixed solution to below 60 ℃, then slowly transferring the feed liquid into 22L ice water, and stirring at room temperature for about 0.5 hour; and (4) carrying out suction filtration, and leaching a filter cake with a small amount of water to obtain a crude product.
Refining: and (3) putting the crude product into a four-mouth bottle, adding 17 g of ethylene glycol monomethyl ether, stirring at 25 +/-5 ℃ for 2 hours, carrying out suction filtration, eluting a filter cake with a proper amount of ethylene glycol monomethyl ether, drying the obtained filter cake with hot air (50 +/-5 ℃) to obtain 11.5 g of brown solid, namely a substance A1-5 (the molecular weight is 430.9, 0.027mol), the purity is 98%, and the yield is 64.5%.
(6) Preparation of Compound A
10g of substance A1-5 (molecular weight of 430.9, 0.023mol) and 0.2L of hydrochloric acid are added into a flask, 10g of aqueous solution of sodium nitrite (containing 3g of sodium nitrite and 0.043mol) is dropwise added under ice bath, reaction is carried out for 1h at room temperature, reaction liquid is slowly poured into 1L of water to precipitate yellow solid, the mixture is stirred for 1h, and the mixture is filtered and dried to obtain 4.0g of compound A (molecular weight of 395.4, 0.01mol), the purity is 98 percent and the yield is 43.1 percent.
Third part
See FIG. 7-FIG. 10, this section is by infrared spectroscopy, nuclear magnetic resonance hydrogen spectroscopy: (1H-NMR) and nuclear magnetic resonance carbon Spectroscopy (C13C-NMR), mass spectrum of the compound prepared in example 1 was examined to verify its structure. Wherein the structural characterization includes, but is not limited to, IR, UV, HRMS, H-NMR, C-NMR.
(1) Infrared spectrum (IR)
The model of the testing instrument: PE spectrum two; test unit: shanghai Zhangjiang grain public service platform, Inc.; the sample preparation method comprises the following steps: the ATR method.
With reference to fig. 7 and table 1, the infrared spectroscopy results show that the sample has significant NH, S ═ O and heteroaromatic rings, and therefore the infrared spectroscopy data of the sample correspond to the molecular structure of the compound.
Infrared spectral data for the compounds of Table 1
| Absorption Peak (cm)-1) | Absorption intensity | Type of vibration | Radical (I) |
| 3196.76 | W | Telescopic vibration | N-H |
| 3021.89 | W | Telescopic vibration | C-H |
| 1637.58～1385.90 | W | Telescopic vibration | Benzene heterocycles |
| 1169.39 | S | Asymmetric telescopic vibration | S＝O |
| 1055 | S | Symmetric telescopic vibration | S＝O |
(2) Nuclear magnetic resonance hydrogen spectrum (1H-NMR) and nuclear magnetic resonance carbon Spectroscopy (C13C-NMR)
A deuterated reagent: about 9 mg of Compound A prepared in example dissolved in 0.50 mL of DMSO-d 6; each group of the compound may be numbered
a) Magnetic resonance hydrogen spectrum (1H-NMR)
Hydrogen spectroscopic data for compounds of table 2
| Serial number | Chemical shift δH(ppm) | Multiplicity J[Hz] | Number of protons | 1H-1H COSY |
| 1 | 2.93 | |
3 | 2 |
| 2 | 8.56 | S, |
1 | 1 |
| 3 | / | / | / | / |
| 4 | / | / | / | / |
| 5 | / | / | / | / |
| 6 | 2.51 | |
3 | / |
| 7 | / | / | / | / |
| 8 | / | / | / | / |
| 9 | 8.47 | d,3.2 | 1 | / |
| 10 | / | / | / | / |
| 11 | 9.61 | S, |
1 | / |
| 12 | / | / | / | / |
| 13 | 7.99 | |
1 | 17,15 |
| 14 | / | / | / | / |
| 15 | 7.22-7.23 | |
1 | 16,13,17 |
| 16 | 7.22-7.23 | |
1 | 15,17 |
| 17 | 7.66-7.69 | |
1 | 16,15,13 |
| 18 | / | / | / | / |
b) Magnetic resonance hydrogen spectrum nuclear magnetic resonance carbon spectrum (13C-NMR)
TABLE 3 carbon Spectroscopy data for Compounds
| Serial number | Chemical shift δC(ppm) | Number of carbon atoms | 13C- | HSQC | HMBC | |
| 1 | 31.2 | 1 | |
1 | / | |
| 2 | / | / | / | / | / | |
| 3 | 170.2 | 1 | Season C | / | 1 | |
| 4 | 109.8，109.9 | 1 | Season C | / | 6 | |
| 5 | 151.8 | 1 | Season C | / | 6 | |
| 6 | 18.0 | 1 | |
6 | / | |
| 7 | 146.3,146.2 | 1 | Season C | / | 9 | |
| 8 | 148.6,146.1 | 1 | Season C | / | 9 | |
| 9 | 146.0,145.8 | 1 | |
9 | / | |
| 10 | 155.9 | 1 | Season C | / | 9 | |
| 11 | / | / | / | / | / | |
| 12 | 148.6or 139.8 | 1 | Season C | / | 16 | |
| 13 | 116.3 | 1 | CH | 13 | / | |
| 14 | 148.6or 139.8 | 1 | Season C | / | 16 | |
| 15 | 118.8 | 1 | CH | 15 | / | |
| 16 | 127.6 | 1 | CH | 16 | / | |
| 17 | 118.8 | 1 | CH | 17 | / | |
| 18 | / | / | / | / | / |
(3) Mass spectrometry
Mass spectral data for the compounds of Table 4
| Mass to charge ratio (m/z) | Remarks for note |
| 396.1 | [M+H]+ |
The mass spectrum results of the compounds showed that the measured molecular weight of 396.0591(M +1) in the HRMS analysis was consistent with the theoretical molecular weight of 396.0595(M +1), and the molecular formula C was deduced from the measured molecular weight15H15FN5O3S2With compounds of formula C15H14FN5O3S2+ H are identical. Mass spectrum of the compound shows that the molecular weight is 395, and integrated Infrared (IR) spectrum, nuclear magnetic resonance hydrogen spectrum (A), (B), (C) and (D)1H-NMR), carbon spectrum (13C-NMR), it can be confirmed that the molecular structure of the sample is consistent with that of the compound.
In conclusion, the compound and the preparation process thereof, which are disclosed by the invention, are applied to a sixth reaction by taking the substance A1-5, hydrochloric acid and a sodium nitrite solution as main raw materials to prepare the compound, and can be used as a standard substance for medicinal compound analysis. The compound is one of main impurities in the synthesis process of LS007, and has important significance for the quality control of LS 007. The compound is prepared by six reactions in total and taking N-methylthiourea, pyridine and 3-chloro-2, 4-pentanedione as initial raw materials, and can be used as a system applicability detection standard. In light of the foregoing description of the preferred embodiment of the present invention, many modifications and variations will be apparent to those skilled in the art without departing from the spirit and scope of the invention. The technical scope of the present invention is not limited to the content of the specification, and must be determined according to the scope of the claims.
Claims (9)
1. A process for preparing LS007 impurity compound A, which is characterized by comprising the following steps:
taking a substance A1-5, hydrochloric acid and a sodium nitrite solution as main raw materials to carry out a sixth reaction; wherein
The structural formula of the substance A1-5 is shown in the specification
2. The production process according to claim 1,
the sixth reaction comprises:
dropwise adding a substance A1-5 and hydrochloric acid into a sodium nitrite solution in an ice bath, and reacting at room temperature to obtain the compound; the molar ratio of A1-5 to sodium nitrite is 1: 2-3;
the reaction formula of the sixth reaction is
3. The production process according to claim 1,
the preparation method of the substance A1-5 comprises the following steps:
taking a substance A1-3, a substance A1-4 and ethylene glycol monomethyl ether as main raw materials to carry out a fifth reaction; wherein
The structural formula of the substance A1-3 is shown in the specification
The structural formula of the substance A1-4 is shown in the specification
The fifth reaction comprises:
mixing and stirring the substance A1-3, the substance A1-4 and ethylene glycol monomethyl ether;
heating to reflux and reacting to obtain a substance A1-5; the molar ratio of the substance A1-3 to the substance A1-4 is 1:1 to 1.2;
the reaction formula of the fifth reaction is:
4. the production process according to claim 3,
the preparation method of the substance A1-4 comprises the following steps:
performing a fourth reaction by taking m-aminobenzenesulfonamide, cyanamide and trimethylchlorosilane as main raw materials;
the fourth reaction comprises:
mixing and stirring m-aminobenzenesulfonamide, cyanamide and a solvent;
cooling to 0 +/-10 ℃ and dropwise adding trimethylchlorosilane;
heating to reflux, and keeping the temperature to perform reaction to obtain a substance A1-4; the molar ratio of the m-aminobenzenesulfonamide to the cyanamide to the acetonitrile is 1: 1.0-1.2.
The reaction formula of the fourth reaction is
5. The production process according to claim 3,
the preparation method of the substance A1-3 comprises the following steps:
carrying out a third reaction by taking a substance A1-2 and a fluorine reagent as main raw materials; wherein
The structural formula of the substance A1-2 is shown in the specification
The third reaction comprises:
mixing the substance A1-2 with a solvent and stirring; cooling to-5 +/-10 ℃, and adding a fluorine reagent selectfluor for reaction; adding ammonia methanol solution, and reacting under continuous stirring to obtain substance A1-3; the molar ratio of the A1-2 to the fluorine reagent and the ammonia methanol is 1: 1-1.5: 1-5; the reaction formula of the third reaction is:
6. the production process according to claim 5,
the preparation method of the substance A1-2 comprises the following steps:
carrying out a second reaction by taking the substance A1-1, N-dimethylformamide dimethyl acetal and xylene as main raw materials; wherein
The structural formula of the substance A1-1 is shown in the specification
The second reaction comprises:
mixing and stirring the substance A1-1, N-dimethylformamide dimethyl acetal and a solvent; heating to reflux, and keeping the temperature for reaction; obtaining the substance A1-2; the A1-2: the molar ratio of N, N-dimethylformamide dimethyl acetal is 1: 1.5 to 3;
the second reaction has the formula
7. The production process according to claim 4,
the preparation method of the substance A1-1 comprises the following steps:
carrying out a first reaction by taking N-methylthiourea, methanol, pyridine and 3-chloro-2, 4-pentanedione as main raw materials; wherein
The first reaction comprises:
mixing and stirring N-methylthiourea and a solvent; adding pyridine to form a white suspension; cooling to 0-5 ℃, and dropwise adding 3-chloro-2, 4-pentanedione; preserving the temperature and carrying out reaction to obtain a substance A1-1; the N-methyl thiourea: pyridine: the ratio of 3-chloro-2, 4-pentanedione is 1: 1-1.2: 1-1.5;
the reaction formula of the first reaction is
8. A compound characterized by having, in a first aspect,
the structural formula of the compound is
9. Use of a compound according to claim 8 as a standard for the analysis of a pharmaceutical compound.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110755792.3A CN114276341A (en) | 2021-07-05 | 2021-07-05 | LS007 impurity compound A and preparation process and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110755792.3A CN114276341A (en) | 2021-07-05 | 2021-07-05 | LS007 impurity compound A and preparation process and application thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN114276341A true CN114276341A (en) | 2022-04-05 |
Family
ID=80868338
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202110755792.3A Pending CN114276341A (en) | 2021-07-05 | 2021-07-05 | LS007 impurity compound A and preparation process and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN114276341A (en) |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103373994A (en) * | 2012-04-19 | 2013-10-30 | 癌症研究技术有限公司 | Therapeutic compounds |
| US20170173021A1 (en) * | 2014-04-04 | 2017-06-22 | Memorial Sloan-Kettering Cancer Center | Method and kits for identifying of cdk9 inhibitors for the treatment of cancer |
| CN107118207A (en) * | 2017-05-22 | 2017-09-01 | 苏州东南药业股份有限公司 | The preparation method of one class CDK inhibitor |
| CN108658966A (en) * | 2018-06-20 | 2018-10-16 | 常州千红生化制药股份有限公司 | The tartrate and its crystal form of selective CDK9 inhibitor |
-
2021
- 2021-07-05 CN CN202110755792.3A patent/CN114276341A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103373994A (en) * | 2012-04-19 | 2013-10-30 | 癌症研究技术有限公司 | Therapeutic compounds |
| US20150105413A1 (en) * | 2012-04-19 | 2015-04-16 | Changzhou Le Sun Pharmaceuticals Limited | Therapeutic compounds |
| US20170173021A1 (en) * | 2014-04-04 | 2017-06-22 | Memorial Sloan-Kettering Cancer Center | Method and kits for identifying of cdk9 inhibitors for the treatment of cancer |
| CN107118207A (en) * | 2017-05-22 | 2017-09-01 | 苏州东南药业股份有限公司 | The preparation method of one class CDK inhibitor |
| CN108658966A (en) * | 2018-06-20 | 2018-10-16 | 常州千红生化制药股份有限公司 | The tartrate and its crystal form of selective CDK9 inhibitor |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN108658966B (en) | Tartrate salts of selective CDK9 inhibitors and crystalline forms thereof | |
| CA2973949A1 (en) | Compounds for improving mrna splicing | |
| CN108383846A (en) | Ba Ruike is for Buddhist nun's A crystal forms and preparation method thereof | |
| CN108147995A (en) | A kind of low 1,8- Naphthalamide derivatives of toxicity and its synthetic method and application | |
| CN106008468B (en) | Bo Maxini A crystal form, B crystal form, C crystal form and preparation method thereof | |
| CN112125911B (en) | CDK9 inhibitor and preparation method and application thereof | |
| CN108164463A (en) | A kind of 1,8- Naphthalamide derivatives and its synthetic method and application with non-small cell lung cancer selective inhibitory | |
| CN114276341A (en) | LS007 impurity compound A and preparation process and application thereof | |
| CN111433213B (en) | Salt forms and crystalline forms thereof as Akt inhibitors | |
| CN110885332B (en) | PDE delta protein degradation targeting chimera and preparation method and application thereof | |
| CN114349747A (en) | Preparation process of LS007 impurity compound A | |
| CN113646312B (en) | Solid BRD4 inhibitor compound and preparation method and application thereof | |
| CN110078706B (en) | Imatinib derivative and preparation method and application thereof | |
| CN110256465B (en) | 2, 4-diaminopyrimidine containing dihydropyranothiazole and application thereof | |
| CN107827877B (en) | Dialkylamino quinazoline compound and application thereof in preparation of antitumor drugs | |
| CN109020883B (en) | 3-cyano-2-aminopyridine derivative and preparation method and application thereof | |
| CN101479278B (en) | Novel 4-amino-3-(-azolyl-phenoxymethyl)-thieno[3,2]pyridine-7-carboxylic acid derivatives | |
| CN109694358B (en) | 2-p-nitrostyryl-4-substituted aminoquinazoline derivative and preparation method and application thereof | |
| CN109180702A (en) | A kind of Thienopyrimidine ketone compound and application thereof | |
| EP3632912A1 (en) | Pyridoquinazoline derivatives useful as protein kinase inhibitors | |
| CN115843298B (en) | Salts and crystal forms of dihydropyrido [2,3-d ] pyrimidinone derivative | |
| CN101497640B (en) | Novel crystal form of clofarabine | |
| CN115947715B (en) | 2, 5-diamino-3, 4-thiophene dicarboxylic acid diethyl ester derivative and application thereof | |
| CN108658945A (en) | A kind of Antitubulin（VDA-1）A crystal forms | |
| CN116685588A (en) | Crystal form of pyridopyrrole compound, preparation method and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination |