CN115611867B - (1, 1-Trichloro-2) carbamate derivative and preparation method and application thereof - Google Patents

(1, 1-Trichloro-2) carbamate derivative and preparation method and application thereof Download PDF

Info

Publication number
CN115611867B
CN115611867B CN202211387962.8A CN202211387962A CN115611867B CN 115611867 B CN115611867 B CN 115611867B CN 202211387962 A CN202211387962 A CN 202211387962A CN 115611867 B CN115611867 B CN 115611867B
Authority
CN
China
Prior art keywords
trichloro
ethyl
dmso
nmr
carbamate
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
Application number
CN202211387962.8A
Other languages
Chinese (zh)
Other versions
CN115611867A (en
Inventor
陈卓
胡高云
李乾斌
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Central South University
Original Assignee
Central South University
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Application filed by Central South University filed Critical Central South University
Priority to CN202211387962.8A priority Critical patent/CN115611867B/en
Publication of CN115611867A publication Critical patent/CN115611867A/en
Application granted granted Critical
Publication of CN115611867B publication Critical patent/CN115611867B/en
Active legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D403/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00
    • C07D403/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings
    • C07D403/12—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings linked by a chain containing hetero atoms as chain links
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
    • C07D401/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings
    • C07D401/12—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings linked by a chain containing hetero atoms as chain links

Landscapes

  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Health & Medical Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)

Abstract

The invention relates to (1, 1-trichloro-2) carbamate derivatives, a preparation method thereof and related medical application. In order to achieve the aim, the invention provides a (1, 1-trichloro-2) carbamate derivative, the structural general formula of which is shown as the formula (I):

Description

(1, 1-Trichloro-2) carbamate derivative and preparation method and application thereof
Technical Field
The invention relates to (1, 1-trichloro-2) carbamate derivatives, a preparation method thereof and related medical application.
Background
Malignant tumor is one of the main causes of death in most countries in the world today, and seriously threatens human health. Investigation shows that the incidence of malignant tumors tends to rise year by year. With the rapid development of molecular biology, scientists have more in-depth and comprehensive knowledge of signal paths and targets related to malignant tumor occurrence and development, and important achievements have been achieved in designing antitumor drugs based on the signal paths and targets. With the wide acceptance of the concept that malignant tumors are a type of cell cycle diseases, anti-tumor drug research aiming at cell cycle regulation becomes a hotspot.
Cell cycle imbalance is a common feature of human cancers, and mutations accumulated in tumor cells can lead to abnormal proliferation, genomic instability and chromosomal instability. Based on this phenomenon, many drugs have been developed to inhibit different cell cycle phases for cancer treatment. Inhibition of the initial stages of the cell cycle may result in viable resting cells, while targeting mitosis has several possibilities to kill cancer cells. Targeting mitosis is a long-term validated method, and currently widely studied are novel drugs targeting mitotic spindle microtubule elements and targeting mitotic non-microtubule effectors, where drugs affecting the mitotic spindle are recognized components in many cancer treatment regimens.
In general, the cell mitosis process is complicated and carefully regulated and eventually divides the replicated genome into two identical daughter cells. Mitosis is divided into five distinct phases: early, mid-late and end. The prophase transition from G2 to mitosis is accompanied by chromosome condensation and onset of nuclear envelope breakdown. In the premetallic stage, chromosomal attachment points attach to spindle microtubules via a search and capture mechanism. In metaphase, chromosomes converge on the equatorial plate equidistant from the centrosome. The centromere microtubules become shorter later, and sister chromatids are separated and moved to opposite poles, which is called segregation. Finally, the cells were bisected during terminal and cytoplasmic division to form two daughter cells (FIG. 1). Numerous small molecule compounds have been designed and studied for mitotic phase related proteins.
Anti-microtubule drugs are the first therapeutic approach to target mitosis, microtubules play an important role in maintaining cell morphology, cell division, signal transduction, and substance delivery. Microtubules polymerize into spindles in the pre-mitosis stage, and the spindles draw chromosomes to move into two autologous cells to the two poles in mitosis, so that cell proliferation is completed. Microtubules play an extremely important role in cell division, have become one of the important targets of anticancer drugs, and microtubule inhibitors have been clinically demonstrated to have remarkable therapeutic effects on various malignant tumors. Microtubule polymerization agents (including paclitaxel and docetaxel) and microtubule depolymerizing agents (including vinorelbine) target primary tubulin, disrupting the kinetic stability of microtubule polymerization-depolymerization, thereby blocking the mitotic cell cycle and inducing apoptosis of tumors. However, these drugs act not only on proliferating tumor cells, but also show significant side effects on normal cells that do not proliferate abnormally. Furthermore, expression of multidrug resistance (MDR) proteins and tubulin isoforms (e.g. mutations in tubulin) are also associated with microtubule inhibiting classes of drugs.
Thus, the identification of novel mitotic drug targets other than tubulin has recently attracted considerable attention. In recent years, several mitotic targets have been explored, including drugs targeting microtubules, kinases, motor proteins and polyprotein complexes. Wherein ubiquitin-proteasome (UPS) pathway mediated ubiquitination of the substrate and its subsequent orderly degradation process is throughout the plasma membrane system of whole cells, is essential for the cells to undergo mitosis throughout the cell cycle, and is of great importance in the treatment of various diseases caused by disturbances of the ubiquitin system, in particular malignant tumors. In general, the ubiquitination process requires the co-participation of ubiquitin activating enzyme (ubiquitin-activatingenzyme, E1), ubiquitin binding enzyme (ubiquitin-conjugatingenzyme, E2) and ubiquitin ligase (ubiquitinligase, E3). In this process, ubiquitin, a highly conserved 76 residue protein, is first linked to ubiquitin activating enzyme (E1) in an ATP-powered reaction. The activated ubiquitin is then transferred to a small ubiquitin binding enzyme (E2) forming a thioester linked E2-ubiquitin intermediate (E2-Ub). E2 acts alone or in combination with E3 ubiquitin ligase to conjugate ubiquitin to the epsilon-amino group of lysine residues in the base protein to form an isopeptide bond. Strict control of these seemingly simple sequential actions of E1-E2-E3 allows for accurate and proper timed ubiquitination/proteolysis.
And the specificity of the ubiquitination system degradation protein is determined by ubiquitin ligase E3. Wherein the late-stage cell division promoting complex (Anaphase Promoting Complex, APC) is the largest E3 ubiquitin ligase, consisting of at least 14 subunits and one coactivator, and forms two different E3 ubiquitin ligase complexes APC Cdc20 or APC Cdh1 (fig. 2) upon activation of cyclin 20 (Cdc 20) or Cdc20 homolog 1 (Cdh 1), APC Cdc20 disrupts key regulators in the cell growth cycle during the transition phase of the mid-to-late-stage mitosis, regulating cell cycle progression, and APC Cdh1 plays an important role mainly in G1.
More and more researches show that Cdc20 promotes the occurrence and development of tumors, the phenomenon of high expression of Cdc20 is found in many tumors, the expression degree of the Cdc20 has close relation with the pathological degree and prognosis of tumor patients, and the higher the expression level of Cdc20 is, the greater the risk is. In summary, the expression level of Cdc20 is an effective prognostic index of tumor patients, and provides a basis for Cdc20 as a novel anti-tumor drug research and development target. In contrast, another coactivator of APC/C, cdh1, is considered a cancer inhibitor.
Currently, as research into Cdc20 continues to be intensive, more and more APC Cdc20 substrates are discovered (table 1), some of which are directly or indirectly involved in the cell division process, cdc20 being able to regulate the ubiquitination or degradation processes of these substrates and thus the cell cycle progression, and therefore more APC Cdc20 substrates are found to help to understand the molecular mechanism of Cdc20 in regulating cell cycle progression.
TABLE 1APCCdc downstream substrates and their function
Inhibition of the Cdc20 pathway using compounds is a new idea for the treatment of tumors. The Cdc20 targeted inhibitor can directly act on Cdc20 to inhibit the activity of the APC Cdc20 complex. Apcin is the only currently reported Cdc 20-specific inhibitor that acts on the WD40 region at the N-terminus of Cdc20, blocking Cdc20 binding to downstream substrates, inhibiting the ubiquitination and degradation processes of downstream substrates (fig. 3). In addition, many compounds have been reported to indirectly affect the Cdc20 pathway and exert antitumor effects. In addition, the active ingredients in certain natural products also have the effect of modulating the Cdc20 pathway (Table 2)
TABLE 2 Structure and function of partial Cdc20 inhibitors
The premature withdrawal of cells from mitosis by mitotic slippage is considered one of the major mechanisms by which cells develop resistance to mitotic drugs. Both mechanisms of action of microtubule inhibitors activate SAC by drug binding to vinca, taxane or colchicine sites, producing unconnected centromeres, thus mitotic cell death or mitotic slippage is observed.
Whereas in the normal cell cycle, mitotic exit is driven by APC/C-Cdc20 dependent cyclin B1 degradation. Inhibition of mitotic progression triggers SAC and inhibits APC/C-Cdc20. However, this inhibition is transient and slow, and cyclin B1 may gradually degrade despite the failure of the mitotic checkpoint. Cells that escape mitotic cell death may die at a later cell cycle stage, stay in the tetraploid state or undergo several rounds of division. Thus, it is envisioned that complete blocking of mitotic exit may be an effective strategy for inducing mitotic cell death. Inhibition of cyclin B1 degradation in vivo by gene elimination of Cdc20 has been demonstrated to be more effective in killing tumor cells than compounds assembled with traditional targeting spindles. On the other hand, classical microtubule-inhibiting antimitotic drugs (vincristine or paclitaxel) induce only partial response in invasive tumors, cdc20 excision can lead to complete arrest of the division phase, leading to massive apoptotic cell death and complete elimination of the tumor in vivo. Meanwhile, studies have shown that in vitro Cdc 20-free tumor cells cannot swing out of metaphase arrest within 6-30 hours and die from mitotic cell death. Importantly, mitotic exit inhibition also affects pRb-null, p53-null or SAC defective cells, indicating its broad utility, making up for the lack of poor therapeutic efficacy of microtubule inhibitors on SAC inactivated cells. Overall, these results indicate that targeting Cdc20 to inhibit Cyclin B1 degradation may be very effective in killing tumor cells.
Thus, designing a small molecule inhibitor that targets both Cdc20 inhibiting mitotic exit and tubulin activating SAC is an effective method of triggering mitotic cell death. The purpose of the mitotic exit inhibitor is to prevent the consumption of cyclin B, resulting in the cells being arrested and dying during mitosis. One preclinical study demonstrated that knockout of Cdc20 was more effective in killing resistant cells than paclitaxel treatment. Another study used APC inhibitor proTAME in combination with either paclitaxel or MLN-8054 anti-mitotic drugs, resulted in activation of apoptosis in cancer cells. By reviewing the clinical trials of antimitotic agents, it is clear that better cancer treatment results can be obtained when using engineered dual target inhibitors.
Current anti-mitotic strategies focus on SAC activation, the main cancer treatment approach is to activate SAC and induce apoptosis using microtubule drugs, but its effectiveness is limited by background Cyclin B1 degradation and mitotic slippage due to residual APC/C-Cdc20 activity. In particular cancer cells, show high mitotic glide rates due to mutations that disrupt SAC, making the treatment fail. Thus, compounds directed against factors downstream of checkpoints, such as Cdc20, are more effective in killing cells, particularly in cells that are prone to slipping and apoptosis, than other mitotic inhibitors, including MIA and kinesin inhibitors.
Apcin as the only CDC20 specific inhibitor reported at present, its antitumor activity is weak, and there is no way to apply clinically, possibly related to the broader protein-protein interaction of CDC20 with a substrate.
At present, no anti-tumor compound aiming at the dual functions of CDC20 and microtubule network is reported, and the dual-function strategy can reduce the nuclear replication, prevent the proteolysis of background Cyclin B1, thereby enhancing the blocking effect of mitosis, simultaneously effectively preventing the occurrence of mitosis slipping, improving the sensitivity of cancer cells to medicines, and opening up a new way for the development of a cancer treatment method based on microtubule inhibitors.
Disclosure of Invention
The invention aims at providing a novel (1, 1-trichloro-2) carbamate derivative, a preparation method of the compound and application of the compound as a possible anti-liver cancer drug.
In order to achieve the above purpose, the technical scheme of the invention is as follows:
A (1, 1-trichloro-2) carbamate derivative has a structural general formula shown in the formula (I-III):
n is 0-1;
X is O;
Wherein R 1 is selected from one of hydrogen, aromatic ring, substituted aromatic ring, aromatic heterocycle, substituted aromatic heterocycle, aliphatic heterocycle, substituted aliphatic heterocycle, C1-C4 open chain amino, C1-C6 straight or branched chain alkoxy, C1-C6 straight or branched chain alkane, C3-C6 aliphatic ring and substituted C3-C6 aliphatic ring, aliphatic lenalidomide substituted C1-C8 straight or branched chain alkyl, and lenalidomide substituted C1-C8 straight or branched chain alkoxyalkyl.
In the structural formula I, R 2,R3,R4 is independently selected from hydrogen, amino, hydroxyl, halogen, C1-C3 alkyl, C1-C3 alkoxy, C1-C3 haloalkyl, C1-C3 alkylamino and phenyl of quinoline ring; y is selected from N, CH; when Y is N, R2, R3 and R4 are not hydrogen at the same time;
In the structural formulas II-III, Y is selected from N and CH; r 5 and R 6 are independently selected from amino, H, halogen, C 1-C8 alkyl, C 1-C8 alkoxy, C 1-C8 haloalkyl, C 1-C8 alkylamino,
Preferably, the aromatic ring is selected from benzene, naphthalene and anthracene; the aromatic heterocycle is selected from pyrrole, furan, thiophene, imidazole, thiazole, oxazole, pyrazole, isoxazole, thiadiazole, oxadiazole, triazole, tetrazole, pyridine, pyrimidine, pyrazine, pyridazine, purine, quinoline, isoquinoline, indole, acridine and carbazole; the aliphatic heterocyclic ring is selected from pyrrolidine, piperazine, piperidine, morpholine and tetrahydrofuran; the substituent groups in the substituted aromatic ring, the substituted aromatic heterocyclic ring and the substituted aliphatic heterocyclic ring are selected from C1-C6 straight-chain or branched-chain alkoxy, C1-C6 straight-chain or branched-chain alkyl, C1-C6 hydroxyalkyl, hydroxyl, amino, nitro, halogenated alkyl and halogen.
Preferably, the substituents in the substituted aromatic ring, the substituted aromatic heterocyclic ring and the substituted aliphatic heterocyclic ring are selected from methyl, ethyl, propyl, isopropyl, methoxy, hydroxyethyl, hydroxyl, amino, nitro, trifluoromethyl and halogen.
Preferably, the structural general formula of the (1, 1-trichloro-2) carbamate derivative is shown as formula I:
wherein n is 1;
X is O;
R 1 is selected from one of hydrogen, C1-C4 straight-chain or branched-chain alkoxy, C1-C4 straight-chain or branched-chain alkane, C3-C6 aliphatic ring, substituted C3-C6 aliphatic ring, benzene ring, substituted benzene ring, aromatic heterocycle, substituted aromatic heterocycle, aliphatic heterocycle and substituted aliphatic heterocycle;
The aromatic heterocycle is selected from pyrrole, imidazole, thiazole, oxazole, pyrazole, pyridine, pyrimidine and indole;
The aliphatic heterocycle is selected from pyrrolidine, piperazine, piperidine and morpholine;
The substituent groups in the substituted aromatic ring, the substituted aromatic heterocyclic ring and the substituted aliphatic heterocyclic ring are selected from methyl, ethyl, methoxy, hydroxyethyl, amino, nitro, trifluoromethyl and halogen;
R 2,R3,R4 is selected from amino, H, =o, halogen, C 1-C3 alkyl, C 1-C3 alkoxy, phenyl moiety of the quinoline ring; y is selected from N, CH; r 2,R3,R4 is not simultaneously hydrogen.
Preferably, the structural general formula of the (1, 1-trichloro-2) carbamate derivative is shown as formula II:
Wherein n is 0-1;
X is O;
R 1 is selected from one of C1-C4 straight-chain or branched-chain alkoxy, C1-C4 straight-chain or branched-chain alkane, C3-C6 aliphatic ring, substituted C3-C6 aliphatic ring benzene ring, substituted benzene ring, aromatic heterocycle, substituted aromatic heterocycle, aliphatic heterocycle and substituted aliphatic heterocycle.
The aromatic heterocycle is selected from pyrrole, imidazole, thiazole, oxazole, pyrazole, pyridine, pyrimidine and indole;
The aliphatic heterocycle is selected from pyrrolidine, piperazine, piperidine and morpholine;
The substituent groups in the substituted aromatic ring, the substituted aromatic heterocyclic ring and the substituted aliphatic heterocyclic ring are selected from methyl, ethyl, methoxy, hydroxyethyl, amino, nitro, trifluoromethyl and halogen;
R 5 and R 6 are selected from amino, H, halogen, C 1-C7 alkyl, C 1-C7 alkoxy, C 1-C7 haloalkyl, C 1-C7 alkylamino,
Y is N, CH.
Preferably, the structural general formula of the (1, 1-trichloro-2) carbamate derivative is shown in formula III:
wherein n is 1;
X is O;
R 1 is selected from one of C1-C4 straight-chain or branched-chain alkoxy, C1-C4 straight-chain or branched-chain alkane, C3-C6 aliphatic ring, substituted C3-C6 aliphatic ring benzene ring, substituted benzene ring, aromatic heterocycle, substituted aromatic heterocycle, aliphatic heterocycle and substituted aliphatic heterocycle.
The aromatic heterocycle is selected from pyrrole, imidazole, thiazole, oxazole, pyrazole, pyridine, pyrimidine and indole;
The aliphatic heterocycle is selected from pyrrolidine, piperazine, piperidine and morpholine;
The substituent groups in the substituted aromatic ring, the substituted aromatic heterocyclic ring and the substituted aliphatic heterocyclic ring are selected from methyl, ethyl, methoxy, hydroxyethyl, amino, nitro, trifluoromethyl and halogen;
R 5 and R 6 are selected from amino, H, halogen, Y is N, CH.
Preferably, the structural formula of the (1, 1-trichloro-2) carbamate derivative is as follows:
The invention also provides a synthesis method of the compounds of the formula (I), the formula (II) and the formula (III), wherein the synthesis route is as follows:
The synthesis steps of the (1, 1-trichloro-2) carbamate derivative comprise:
1. reacting a hydroxyl substituted compound with p-nitrophenyl chloroformate in an anhydrous aprotic solvent to obtain an intermediate;
2. The intermediate reacts with ammonia water in a mixed solvent of halogenated alkane and methanol to obtain a carbamate product;
3. refluxing the carbamate product and chloral hydrate at 80-100 ℃ to obtain a (1, 1-trichloro-2) carbamate part;
4. the (1, 1-trichloro-2) carbamate moiety is first chlorinated by thionyl chloride and then reacted with an amine to give the target compound.
Preferably, the molar ratio of the reaction in the step 1 is that the hydroxy-substituted compound is p-nitrophenyl chloroformate: chloral hydrate=1 (1.2-1.5): 8-12.
Preferably, step 1 is reacted for 4-12 hours to give an intermediate.
Preferably, the reaction in the step 1 can be catalyzed by weak base to accelerate the reaction process.
Preferably, the weak base is any one or combination of triethylamine and diisopropylethylamine, and the molar ratio of the materials is: hydroxy-substituted compound base = 1:1.2-1:1.5.
Preferably, the anhydrous aprotic solvent is dichloromethane.
Preferably, the mass of the haloalkane and the methanol in the mixed solvent of the haloalkane and the methanol in the step 2 is 1:1-5:1.
Preferably, the concentration of ammonia water in the step 2 is 10-35%.
Preferably, step 3 reaction molar feed ratio: urethane, chloral hydrate = 1:8-1:12
Preferably, the reflux time of step 3 is 6-48 hours.
Preferably, the reaction temperature in step 3 is 100 ℃.
Preferably, step 4 is carried out at a temperature of rt-50 ℃.
Preferably, in step 4 (1, 1-trichloro-2) the carbamate moiety (1-fold equivalent) is first dissolved in anhydrous aprotic solvent, chlorinated by thionyl chloride (5-fold equivalent), and then passed through anhydrous dichloromethane: and (3) carrying out substitution reaction on anhydrous acetonitrile (1:1) serving as a mixed solvent and raw material amine (1.2 times of equivalent) to generate a corresponding target compound.
The invention also provides application of the (1, 1-trichloro-2) carbamate derivative in preparing medicines for treating breast cancer, melanoma, lung adenocarcinoma, hepatocellular carcinoma, cervical cancer and ovarian cancer.
The invention is further explained below:
Based on the structure of the cdc20 specific inhibitor apcin, different substituent groups are introduced on pyrimidine rings of the cdc20 specific inhibitor to obtain various derivatives (structural parent nucleus I), and after chlorine atoms are introduced, the compound has more interaction sites with the cdc20, so that the affinity is enhanced, and the inhibition effect of the compound on tumor cells is greatly enhanced.
And after pyrimidine in apcin structure is replaced by purine (structural parent nucleus II) or benzopyrazole (structural parent nucleus III), the inhibition effect on tumor cells is further enhanced. On the basis of maintaining a certain inhibition effect of cdc20, the compound enhances the interaction between the compound and a tubulin dimer due to the introduction of purine or benzopyrazole, and generates a microtubule aggregation inhibition effect. The compound has dual-function anti-tumor effect, and is hopeful to become a novel anti-tumor lead compound.
Compared with the prior art, the invention has the following beneficial effects:
1. the invention enriches the structure of the cdc20 inhibitor on the basis of apcin, and three structural parent nuclei are obtained;
2. Compared with apcin, the tumor inhibition activity of the target compound is greatly enhanced, and the target compound has good inhibition effect in a plurality of tumor cell strains;
3. The invention discovers that the novel compound has good microtubule aggregation inhibition effect besides the original cdc20 inhibition activity of apcin through researching an anti-tumor activity mechanism of the compound, and opens up a new way for searching a treatment method for the double-function cancer aiming at an APC (advanced cancer cell) passage and microtubule inhibition.
Drawings
FIG. 1 is a schematic diagram of the mitotic process and related target proteins and complexes;
FIG. 2 is a graph showing that the binding of the APC/C RING E3 ubiquitin ligase to its coactivator Cdc20/Cdh1 promotes ubiquitination of the substrate;
FIG. 3 is a D-box binding pocket (left) on the side of the WD40 region to which small molecule compound Apcin binds and the small molecule compound TAME interferes with the binding site of the IR tail by mimicking the isoleucine-arginine (IR) tail of Cdc20 and Cdh1 to APC 3;
FIG. 4 shows immunofluorescence assay results of drug-treated HepG2 cell lines;
FIG. 5 shows the results of apoptosis experiments (compound pro-apoptotic fluorescence) for drug-treated HepG2 cell lines.
Detailed Description
The present invention will be described in detail with reference to examples. It should be noted that, without conflict, the embodiments of the present invention and features of the embodiments may be combined with each other.
Example 1
2- (2-Methyl-5-nitro-1H-imidazol-1-yl) -ethyl- (2, 2-trichloro-1- ((4-methylpyrimidine-2-) amino) -ethyl) -carbamic acid ester (5 b)
Step 1
Synthesis of 2- (2-methyl-5-nitro-1H-imidazolin-1-yl) ethylcarbamate
30.0G of metronidazole (0.175 mol) is weighed and put into a 500mL eggplant type bottle, then 300mL of anhydrous dichloromethane is sequentially added for dissolution, 30mL of anhydrous triethylamine (0.210 mol) is added, 42.5g of p-nitrophenyl chloroformate (0.210 mol) is added under the ice bath condition after uniform stirring, and finally 30mL of anhydrous dichloromethane is added for rinsing the bottle mouth. The reaction was stirred overnight at room temperature and TLC monitored the metronidazole reaction to be substantially complete and stopped. 62mL of ammonia (28%) was slowly added to the reaction mixture at 10℃using a constant pressure dropping funnel, and the reaction was completed for 3 hours. The reaction mixture was filtered off to obtain 58.3g of a solid, the obtained solid was dissolved in 200mL of sodium hydroxide solution (1 mol/L), extracted 3 times with 200mL of methylene chloride, the organic layers were combined, concentrated under reduced pressure, and dried to obtain 18.6g of 2- (2-methyl-5-nitro-1H-imidazolin-1-yl) ethylcarbamate as a yellow crystalline compound. Yield in two steps: 50%, melting point: 151.5-153.1 ℃.
Step 2
Synthesis of 2- (2-methyl-5-nitro-1H-imidazolin-1-yl) ethyl (2, 2-trichloro-1-hydroxyethyl) carbamate
77.0G of chloral hydrate (0.467 mol) is weighed and put into a 100mL eggplant type bottle, the bottle is placed on an oil bath pot for heating, after the chloral hydrate is melted, 10.0g of compound 2- (2-methyl-5-nitro-1H-imidazolin-1-yl) ethylcarbamate (0.047 mol) is slowly added for stirring reaction at 100 ℃ for 48 hours, and the reaction is stopped. The reaction solution was cooled to room temperature, 50mL of ethyl acetate was added, and after ultrasonic treatment to form a turbid liquid, suction filtration was performed to obtain 11.6g of a white compound. Yield: 68%, melting point: 168.7-169.7 ℃.
Step 3
The intermediate 2- (2-methyl-5-nitro-1H-imidazolin-1-yl) ethyl (2, 2-trichloro-1-hydroxyethyl) carbamate and 4-methyl-2-aminopyrimidine are used as raw materials to obtain a compound 5b.
The specific operation is as follows: 0.50g of intermediate 2- (2-methyl-5-nitro-1H-imidazoline-1-yl) ethyl (2, 2-trichloro-1-hydroxyethyl) carbamate (1.38 mmol) is weighed into a 50mL eggplant-type bottle, 30mL of anhydrous dichloromethane is measured and added into the reaction bottle, 1 drop of triethylamine is dropwise added, after stirring uniformly, 0.50mL of thionyl chloride (6.91 mmol) is slowly dropwise added into the reaction bottle through a constant pressure dropping funnel under ice bath, after the dropwise addition is finished, the reaction bottle is transferred into an oil bath pot, and reflux reaction is carried out overnight at 40 ℃ under the protection of nitrogen, and the reaction is stopped. After the reaction solution was cooled to room temperature, it was concentrated under reduced pressure, and 20mL of anhydrous dichloromethane was added for sufficient dissolution, and the concentration was repeated 3 times under reduced pressure to remove residual thionyl chloride, to obtain a yellow solid. The resulting yellow solid was dissolved with 15mL of anhydrous dichloromethane and 15mL of anhydrous acetonitrile, another starting amine (1.66 mmol) was added to the reaction solution, and after 24 hours 10mL of methanol was added and stirred for 30min to quench the reaction. The reaction mixture was concentrated under reduced pressure, dissolved in 50mL of methylene chloride, washed 3 times with 50mL of saturated brine, and concentrated under reduced pressure to give a crude product. Dissolving the crude product with 20mL of methanol, mixing with 80-100 mesh silica gel, loading into a column with 200-300 mesh silica gel, purifying by column chromatography, eluting with dichloromethane and methanol solution, and separating and purifying to obtain the target compound.
The title compound was a white solid, yield: 21.42%, melting point ：209.0～209.9℃,HPLC：97.19％.1H NMR(500MHz,DMSO-d6):δ8.27(d,J＝5.0Hz,1H),8.00(s,1H),7.97(d,J＝8.8Hz,1H),6.97(d,J＝9.5Hz,1H),6.73(d,J＝5.0Hz,1H),6.60(t,J＝9.2Hz,1H),4.56–4.46(m,3H),4.35(dd,J＝10.7,5.7Hz,1H),2.39(s,3H),2.31(s,3H).13C NMR(500MHz,DMSO-d6):δ160.69,155.24,152.06,138.88,133.53,112.85,102.88,70.12,63.46,45.90,24.10,14.37.HRMS(ESI)m/z calcd for[C14H16Cl3N7O4+H]+:452.0329,found:452.0401.
Example 2
2- (2-Methyl-5-nitro-1H-imidazol-1-yl) -ethyl- (2, 2-trichloro-1- ((4-methoxypyrimidin-2-) amino) -ethyl) -carbamic acid ester (5 c)
Steps 1 and 2 are the same as in example 1.
The procedure of example 1, step 3, was followed using the intermediates 2- (2-methyl-5-nitro-1H-imidazolin-1-yl) ethyl (2, 2-trichloro-1-hydroxyethyl) carbamate and 4-methoxy-2-aminopyrimidine as starting materials to give compound 5c. Pale yellow solid, yield: 32.66%, melting point ：81.6～83.3℃,HPLC：98.63％.1H NMR(500MHz,DMSO-d6):δ8.14(d,J＝5.7Hz,1H),8.01(d,J＝16.7Hz,2H),7.05(s,1H),6.57(d,J＝13.3Hz,1H),6.28(d,J＝5.7Hz,1H),4.51(dd,J＝17.4,4.3Hz,3H),4.38–4.33(m,1H),3.85(s,3H),2.39(s,3H).13C NMR(500MHz,DMSO-d6):δ170.22,160.87,158.59,155.27,152.06,138.88,133.51,102.59,99.44,70.35,63.48,53.74,45.89,14.36.HRMS(ESI)m/z calcd for[C14H16Cl3N7O5+H]+:468.0278,found:468.0370.
Example 3
2- (2-Methyl-5-nitro-1H-imidazol-1-yl) -ethyl- (2, 2-trichloro-1- ((6-oxo-1, 6-dihydropyrimidine-2-) amino) ethyl) carbamic acid (5 d)
Steps 1 and 2 are the same as in example 1.
Compound 5d was obtained according to the synthesis procedure of example 1, step 3 starting from intermediate 2- (2-methyl-5-nitro-1H-imidazolin-1-yl) ethyl (2, 2-trichloro-1-hydroxyethyl) carbamate and isocytosine. Pale yellow solid, yield: 16.24%, melting point ：192.3～193.2℃,HPLC：98.39％.1H NMR(500MHz,DMSO-d6):δ11.14(s,1H),8.71(d,J＝8.4Hz,1H),8.01(s,1H),7.64(d,J＝6.1Hz,1H),7.11(d,J＝9.3Hz,1H),6.49(t,J＝9.1Hz,1H),5.72(d,J＝6.1Hz,1H),4.55–4.48(m,3H),4.37(dd,J＝7.5,5.0Hz,1H),2.41(s,3H).13C NMR(500MHz,DMSO-d6):δ162.08,155.74,155.43,153.95,152.10,138.89,133.56,105.94,101.66,69.29,63.28,46.00,14.41.HRMS(ESI)m/z calcd for[C13H14Cl3N7O5+H]+:454.0122,found:454.0200.
Example 4
2- (2-Methyl-5-nitro-1H-imidazol-1-yl) -ethyl- (2, 2-trichloro-1- ((4-aminopyrimidin-2-) amino) -ethyl) -carbamic acid ester (5 e)
Steps 1 and 2 are the same as in example 1.
The procedure of example 1, step 3, was followed using the intermediates 2- (2-methyl-5-nitro-1H-imidazolin-1-yl) ethyl (2, 2-trichloro-1-hydroxyethyl) carbamate and 2, 4-diaminopyrimidine as starting materials to give compound 5e. Pale yellow solid, yield: 28.69%, melting point ：104.2～105.0℃,HPLC：98.55％.1H NMR(500MHz,DMSO-d6):δ8.02(s,1H),7.95(d,J＝8.5Hz,1H),7.74(d,J＝5.3Hz,1H),6.64(s,2H),6.53(t,J＝9.0Hz,1H),6.15(s,1H),5.87(d,J＝5.1Hz,1H),4.56-4.46(m,3H),4.32(d,J＝11.4Hz,1H),2.40(s,3H).13C NMR(500MHz,DMSO-d6):δ164.54,160.64,156.04,155.13,152.07,138.88,133.52,103.38,97.97,69.98,63.36,45.92,14.37.HRMS(ESI)m/z calcd for[C13H15Cl3N8O4+H]+:453.0282,found:453.0360.
Example 5
2- (2-Methyl-5-nitro-1H-imidazol-1-yl) -ethyl- (2, 2-trichloro-1- ((4-chloropyrimidin-2-) amino) -ethyl) -carbamic acid ester (5 f)
Steps 1 and 2 are the same as in example 1.
The procedure of example 1, step 3, was followed using the intermediates 2- (2-methyl-5-nitro-1H-imidazolin-1-yl) ethyl (2, 2-trichloro-1-hydroxyethyl) carbamate and 4-chloro-2-aminopyrimidine as starting materials to give compound 5f. White solid, yield: 20.18%, melting point ：182.3～183.4℃,HPLC：98.75％.1H NMR(500MHz,DMSO-d6):δ8.40(d,J＝4.8Hz,1H),8.06(d,J＝8.4Hz,1H),7.99(s,1H),7.87(s,1H),6.98(d,J＝5.0Hz,1H),6.55(s,1H),4.56–4.47(m,3H),4.40–4.35(m,1H),2.41(s,3H).13C NMR(500MHz,DMSO-d6):δ161.18,155.35,152.08,138.90,133.51,112.61,102.13,70.30,63.51,45.88,14.41.HRMS(ESI)m/z calcd for[C13H13Cl4N7O4+H]+:471.9783,found:471.9864.
Example 6
2- (2-Methyl-5-nitro-1H-imidazol-1-yl) -ethyl- (2, 2-trichloro-1- ((4-trifluoromethyl pyrimidine-2-) amino) -ethyl) -carbamic acid ester (5 g)
Steps 1 and 2 are the same as in example 1.
The procedure is as in step 3 of example 1, starting from intermediate 2- (2-methyl-5-nitro-1H-imidazolin-1-yl) ethyl (2, 2-trichloro-1-hydroxyethyl) carbamate and 4-trifluoromethyl-2-aminopyrimidine to give compound 5g. Yellow oil, yield ：5.26％,HPLC：97.81％.1H NMR(500MHz,DMSO-d6):δ8.79(d,J＝4.4Hz,1H),8.12(s,1H),8.03(d,J＝8.6Hz,1H),7.98(s,1H),7.28(d,J＝4.9Hz,1H),6.61(s,1H),4.57–4.46(m,3H),4.40–4.34(m,1H),2.40(s,3H).HRMS(ESI)m/z calcd for[C14H13Cl3F3N7O4+H]+:506.0047,found:506.0140.
Example 7
2- (2-Methyl-5-nitro-1H-imidazol-1-yl) -ethyl- (2, 2-trichloro-1- ((pyrimidine-4-) amino) -ethyl) -carbamic acid ester (6 a)
Steps 1 and 2 are the same as in example 1.
Compound 6a was obtained according to the synthesis procedure of example 1, step 3 starting from intermediate 2- (2-methyl-5-nitro-1H-imidazolin-1-yl) ethyl (2, 2-trichloro-1-hydroxyethyl) carbamate and 4-aminopyrimidine. Yellow solid, yield: 32.24%, melting point ：179.4～180.2℃,HPLC：99.79％.1H NMR(500MHz,DMSO-d6):δ8.59–8.48(m,2H),8.21(d,J＝5.9Hz,1H),8.04–7.94(m,2H),6.88(d,J＝5.9Hz,1H),6.77(t,J＝8.6Hz,1H),4.50(d,J＝11.0Hz,3H),4.35(dd,J＝10.7,4.5Hz,1H),2.39(s,3H).13C NMR(500MHz,DMSO-d6):δ161.10,158.30,155.81,155.59,152.14,138.87,133.56,107.09,101.98,68.80,63.24,46.04,14.41.HRMS(ESI)m/z calcd for[C13H14Cl3N7O4+H]+:438.0173,found:438.0264.
Example 8
2- (2-Methyl-5-nitro-1H-imidazol-1-yl) -ethyl- (2, 2-trichloro-1- ((quinolin-2-) amino) -ethyl) -carbamic acid ester (6 b)
Steps 1 and 2 are the same as in example 1.
Compound 6b was obtained according to the synthesis procedure of example 1, step 3 starting from intermediate 2- (2-methyl-5-nitro-1H-imidazolin-1-yl) ethyl (2, 2-trichloro-1-hydroxyethyl) carbamate and 2-aminoquinoline. Dark yellow solid, yield: 25.62%, melting point ：119.9～121.0℃,HPLC：99.75％.1H NMR(500MHz,DMSO-d6):δ8.35(d,J＝8.8Hz,1H),8.03–7.98(m,2H),7.71(d,J＝7.9Hz,1H),7.59–7.54(m,2H),7.49(d,J＝9.2Hz,1H),7.27(t,J＝7.2Hz,1H),7.13(d,J＝8.9Hz,1H),6.94(t,J＝9.0Hz,1H),4.51(dd,J＝11.7,4.8Hz,3H),4.35–4.30(m,1H),2.34(s,3H).13C NMR(500MHz,DMSO-d6):δ155.57,155.16,152.16,147.19,138.82,137.80,133.55,129.84,127.96,126.72,124.23,123.01,113.30,102.69,69.54,63.17,46.07,14.38.HRMS(ESI)m/z calcd for[C18H17Cl3N6O4+H]+:487.0377,found:487.0463.
Example 9
2- (2-Methyl-5-nitro-1H-imidazol-1-yl) -ethyl- (2, 2-trichloro-1- ((isoquinolin-3-) amino-) ethyl) -carbamic acid ester (6 c)
Steps 1 and 2 are the same as in example 1.
Compound 6c was obtained according to the synthesis procedure of example 1, step 3 starting from intermediate 2- (2-methyl-5-nitro-1H-imidazolin-1-yl) ethyl (2, 2-trichloro-1-hydroxyethyl) carbamate and 3-aminoisoquinoline. Yellow solid, yield: 28.32%, melting point ：122.6～124.2℃,HPLC：99.50％.1H NMR(500MHz,DMSO-d6):δ8.96(s,1H),8.27(d,J＝8.7Hz,1H),7.99(s,1H),7.90(d,J＝8.2Hz,1H),7.64(d,J＝8.3Hz,1H),7.58–7.54(m,1H),7.32–7.28(m,1H),7.05(s,1H),6.81–6.72(m,2H),4.49(d,J＝11.5Hz,3H),4.33(d,J＝5.3Hz,1H),2.34(s,3H).13C NMR(500MHz,DMSO-d6):δ155.57,152.99,152.12,151.48,138.86,138.56,133.53,131.03,128.23,125.27,124.18,123.73,103.23,100.17,70.84,63.23,46.02,14.36.HRMS(ESI)m/z calcd for[C18H17Cl3N6O4+H]+:487.0377,found:487.0469.
Example 10
2- (2-Methyl-5-nitro-1H-imidazol-1-yl) -ethyl- (2, 2-trichloro-1- ((isoquinolin-1-) amino) -ethyl) -carbamic acid ester (6 d)
Steps 1 and 2 are the same as in example 1.
Compound 6d was obtained according to the synthesis procedure of example 1, step 3 starting from intermediate 2- (2-methyl-5-nitro-1H-imidazolin-1-yl) ethyl (2, 2-trichloro-1-hydroxyethyl) carbamate and 1-aminoisoquinoline. White solid, yield: 27.43%, melting point ：113.5～114.8℃,HPLC：97.92％.1H NMR(500MHz,DMSO-d6):δ8.06(d,J＝8.4Hz,1H),8.02–7.94(m,2H),7.84(d,J＝8.0Hz,1H),7.78(d,J＝8.7Hz,1H),7.73(t,J＝7.5Hz,1H),7.64(t,J＝7.6Hz,1H),7.35(d,J＝8.8Hz,1H),7.18(d,J＝5.7Hz,1H),7.07(t,J＝8.7Hz,1H),4.49(dd,J＝21.6,11.7Hz,3H),4.37–4.31(m,1H),2.36(s,3H).13C NMR(500MHz,DMSO-d6):δ155.11,152.99,151.98,140.88,138.89,137.32,133.50,130.92,127.44,127.06,122.66,117.67,113.11,103.34,69.26,63.53,45.81,14.34.HRMS(ESI)m/z calcd for[C18H17Cl3N6O4+H]+:487.0377,found:487.0463.
Example 11
2- (2-Methyl-5-nitro-1H-imidazol-1-yl) -ethyl- (1- (1H-benzo [ d ] imidazol-1-yl) -2, 2-trichloroethyl) -carbamic acid ester (6 e)
Steps 1 and 2 are the same as in example 1.
The procedure of example 1, step 3, was followed using the intermediate 2- (2-methyl-5-nitro-1H-imidazolin-1-yl) ethyl (2, 2-trichloro-1-hydroxyethyl) carbamate and benzimidazole as starting materials to give compound 6e. White solid, yield: 22.13%, melting point ：123.6～124.9℃,HPLC：97.97％.1H NMR(500MHz,DMSO-d6):δ9.68(d,J＝9.8Hz,1H),8.63(s,1H),8.00(d,J＝6.1Hz,1H),7.96(t,J＝11.1Hz,1H),7.71(t,J＝7.8Hz,1H),7.33(t,J＝7.4Hz,1H),7.27(t,J＝7.4Hz,1H),6.88(t,J＝14.6Hz,1H),4.60–4.51(m,3H),4.40(dt,J＝8.5,3.8Hz,1H),2.38(s,3H).13C NMR(500MHz,DMSO-d6):δ155.67,152.01,142.82,142.73,138.91,134.14,133.57,123.96,123.03,120.18,112.05,99.86,73.06,63.74,45.86,14.34.HRMS(ESI)m/z calcd for[C16H15Cl3N6O4+H]+:461.0220,found:461.0127.
Example 12
2- (2-Methyl-5-nitro-1H-imidazol-1-yl) -ethyl- (2, 2-trichloro-1- (6- (2, 2-trichloro-1- ((2- (2-methyl-5-nitro-1H-imidazol-1-yl) -ethoxycarbonylamino-) -ethyl) -amino) -9H-purin-9-yl-ethyl) -carbamic acid ester (6 f)
Steps 1 and 2 are the same as in example 1.
Compound 6f was obtained according to the synthesis procedure of example 1, step 3 starting from intermediate 2- (2-methyl-5-nitro-1H-imidazolin-1-yl) ethyl (2, 2-trichloro-1-hydroxyethyl) carbamate and adenine. Dark yellow solid, yield: 10.56%, melting Point ：173.8～175.3℃,HPLC：97.12％.1H NMR(500MHz,DMSO-d6):δ9.63(d,J＝9.3Hz,1H),8.62(s,1H),8.48(s,1H),7.97(s,2H),7.87(d,J＝21.2Hz,2H),6.95(d,J＝10.0Hz,1H),6.90(s,1H),4.58–4.34(m,8H),2.39(s,3H),2.37(s,3H).HRMS(ESI)m/z calcd for[C23H23Cl6N13O8+H]+:819.9924,found:820.0001.
Example 13
2- (2-Methyl-5-nitro-1H-imidazol-1-yl) -ethyl- (2, 2-trichloro-1- ((4, 6-dichloropyrimidin-2-) amino) -ethyl) -carbamic acid ester (7 a)
Steps 1 and 2 are the same as in example 1.
The procedure of example 1, step 3, was followed using the intermediates 2- (2-methyl-5-nitro-1H-imidazolin-1-yl) ethyl (2, 2-trichloro-1-hydroxyethyl) carbamate and 2-amino-4, 6 dichloropyrimidine as starting materials to give compound 7a. Pale yellow solid, yield: 46.23%, melting point ：2175.5～176.3℃,HPLC：99.73％.1H NMR(500MHz,DMSO-d6):δ8.56(d,J＝9.2Hz,1H),8.17(d,J＝9.0Hz,1H),7.98(s,1H),7.21(s,1H),6.45(t,J＝9.1Hz,1H),4.55–4.48(m,3H),4.38(dt,J＝9.5,4.4Hz,1H),2.42(s,3H).13C NMR(500MHz,DMSO-d6):δ160.73,155.42,152.12,138.90,133.50,111.40,101.52,70.47,63.53,45.90,14.45.HRMS(ESI)m/z calcd for[C13H12Cl5N7O4+H]+:505.9393,found:505.9472.
Example 14
2- (2-Methyl-5-nitro-1H-imidazol-1-yl) -ethyl- (2, 2-trichloro-1- ((4, 6-dichloro-5-methylpyrimidine-2-) amino) -ethyl) -carbamic acid ester (7 b)
Steps 1 and 2 are the same as in example 1.
Compound 7b was obtained according to the synthesis procedure of example 1, step 3 starting from the intermediates 2- (2-methyl-5-nitro-1H-imidazolin-1-yl) ethyl (2, 2-trichloro-1-hydroxyethyl) carbamate and 2-amino-4, 6 dichloro-5-methylpyrimidine. White solid, yield: 46.66%, melting point ：140.3～142.1℃,HPLC：99.82％.1H NMR(400MHz,DMSO-d6):δ8.24(d,J＝9.3Hz,1H),8.15(d,J＝9.0Hz,1H),7.99(s,1H),6.40(t,J＝9.1Hz,1H),4.56–4.46(m,3H),4.37(dt,J＝9.3,3.9Hz,1H),2.42(s,3H),2.26(s,3H).13C NMR(400MHz,DMSO-d6):δ158.23,155.43,152.15,138.90,133.55,117.65,101.74,70.55,63.51,45.92,15.41,14.47.HRMS(ESI)m/z calcd for[C14H14Cl5N7O4+H]+:519.9550,found:519.9629.
Example 15
2- (2-Methyl-5-nitro-1H-imidazol-1-yl) -ethyl- (2, 2-trichloro-1- ((5-amino-4, 6-dichloropyrimidin-2-) amino) -ethyl) -carbamic acid ester (7 c)
Steps 1 and 2 are the same as in example 1.
The procedure of example 1, step 3, was followed using the intermediates 2- (2-methyl-5-nitro-1H-imidazolin-1-yl) ethyl (2, 2-trichloro-1-hydroxyethyl) carbamate and 2, 5-diamino-4, 6 dichloropyrimidine as starting materials to give compound 7c. Tan solid, yield: 27.68%, melting point ：92.9～93.7℃,HPLC：99.73％.1H NMR(500MHz,DMSO-d6):δ8.00(d,J＝8.9Hz,1H),7.98(s,1H),7.21(d,J＝9.5Hz,1H),6.26(t,J＝9.2Hz,1H),5.16(s,2H),4.56–4.45(m,3H),4.37(dt,J＝9.6,4.6Hz,1H),2.40(s,3H).13C NMR(500MHz,DMSO-d6):δ155.34,152.07,151.11,145.55,138.88,133.49,129.80,102.40,71.00,63.41,45.92,14.40.HRMS(ESI)m/z calcd for[C13H13Cl5N8O4+H]+:520.9502,found:520.9581.
Example 16
2- (2-Methyl-5-nitro-1H-imidazol-1-yl) -ethyl- (1- ((2-amino-4, 6-dichloropyrimidin-5-) amino) -2, 2-trichloroethyl) -carbamic acid ester (7 d)
Steps 1 and 2 are the same as in example 1.
Compound 7d was obtained according to the synthesis procedure of example 1, step 3 starting from 2- (2-methyl-5-nitro-1H-imidazolin-1-yl) ethyl (2, 2-trichloro-1-hydroxyethyl) carbamate and 2, 5-diamino-4, 6 dichloropyrimidine. White solid, yield: 20.18%, melting point ：125.1～125.7℃,HPLC：99.14％.1H NMR(500MHz,DMSO-d6):δ8.35(s,1H),8.34(d,J＝9.5Hz,1H),8.01(s,1H),7.76–7.74(m,1H),7.26(s,2H),5.58(t,J＝10.0Hz,1H),4.61(d,J＝10.6Hz,1H),4.47(d,J＝11.9Hz,3H),4.42–4.35(m,1H),2.40(s,3H).13C NMR(500MHz,DMSO-d6):δ158.85,156.40,155.83,152.17,138.83,133.53,121.90,102.23,75.27,63.21,46.09,14.45.HRMS(ESI)m/z calcd for[C13H13Cl5N8O4+H]+:520.9502,found:520.9574.
Example 17
2- (2-Methyl-5-nitro-1H-imidazol-1-yl) -ethyl- (2, 2-trichloro-1- ((6-chloro-9H-purin-9-yl) -ethyl) -carbamic acid ester (8 a)
Steps 1 and 2 are the same as in example 1.
Compound 8a was obtained according to the synthesis procedure of example 1, step 3 starting from intermediate 2- (2-methyl-5-nitro-1H-imidazolin-1-yl) ethyl (2, 2-trichloro-1-hydroxyethyl) carbamate and 6-chloropurine. Pale yellow solid, yield: 21.32%, melting point ：101.4～101.8℃,HPLC：96.82％.1H NMR(500MHz,DMSO-d6):δ9.76(d,J＝10.0Hz,1H),8.98(s,1H),8.93(s,1H),7.91(s,1H),7.00(d,J＝10.0Hz,1H),4.57–4.45(m,4H),2.37(s,3H).13C NMR(500MHz,DMSO-d6):δ155.56,153.11,152.38,151.86,150.38,144.65,138.98,133.45,130.31,98.77,71.56,63.87,45.75,14.35.HRMS(ESI)m/z calcd for[C14H12Cl4N8O4+H]+:496.9736,found:496.9782.
Example 18
2- (2-Methyl-5-nitro-1H-imidazol-1-yl) -ethyl- (2, 2-trichloro-1- ((6- (N, N-dimethylamino) -9H-purin-9-yl) -ethyl) -carbamic acid ester (8 b)
Steps 1 and 2 are the same as in example 1.
Compound 8b was obtained according to the synthesis procedure of example 1, step 3 starting from intermediate 2- (2-methyl-5-nitro-1H-imidazolin-1-yl) ethyl (2, 2-trichloro-1-hydroxyethyl) carbamate and 6-dimethylaminopurine. White solid, yield: 20.23%, melting point ：113.6～115.2℃,HPLC：98.22％.1H NMR(500MHz,CDCl3):δ8.51(s,1H),8.31(s,1H),7.96(s,1H),7.94(s,1H),6.65(d,J＝9.7Hz,1H),4.64–4.58(m,3H),4.54–4.49(m,1H),3.58(s,6H),2.47(s,3H).13C NMR(500MHz,DMSO-d6):δ155.52,154.74,153.01,151.93,151.01,138.94,137.06,133.50,118.26,99.65,70.73,63.80,45.81,14.32.HRMS(ESI)m/z calcd for[C16H18Cl3N9O4+H]+:506.0547,found:506.0638.
Example 19
2- (2-Methyl-5-nitro-1H-imidazol-1-yl) -ethyl- (1- (6-benzamide-9H-purin-9-yl) -2, 2-trichloroethyl) -carbamic acid ester (8 c)
Steps 1 and 2 are the same as in example 1.
The procedure of example 1, step 3, was followed using the intermediate 2- (2-methyl-5-nitro-1H-imidazolin-1-yl) ethyl (2, 2-trichloro-1-hydroxyethyl) carbamate and N6-benzoyladenine as starting materials to give compound 8c. White solid, yield: 22.65%, melting point ：183.7～184.9℃,HPLC：97.27％.1H NMR(500MHz,DMSO-d6):δ11.35(s,1H),9.71(d,J＝9.9Hz,1H),8.85(s,1H),8.78(s,1H),8.06(d,J＝7.4Hz,2H),7.97(s,1H),7.66(t,J＝7.2Hz,1H),7.57(t,J＝7.3Hz,2H),7.05(d,J＝9.9Hz,1H),4.58(d,J＝11.0Hz,3H),4.45(s,1H),2.38(s,3H).13C NMR(500MHz,DMSO-d6):δ166.08,155.59,152.94,152.91,151.92,151.30,141.70,138.97,133.61,133.49,133.05,129.01,128.97,124.43,99.26,71.09,63.89,45.78,14.33.HRMS(ESI)m/z calcd for[C21H18Cl3N9O5+H]+:582.0496,found:582.0577.
Example 20
2- (2-Methyl-5-nitro-1H-imidazol-1-yl) -ethyl- (1- (6-benzylamino-9H-purin-9-yl) -2, 2-trichloroethyl) -carbamic acid ester (8 d)
Steps 1 and 2 are the same as in example 1.
Compound 8d was obtained according to the synthesis procedure of example 1, step 3 starting from intermediate 2- (2-methyl-5-nitro-1H-imidazolin-1-yl) ethyl (2, 2-trichloro-1-hydroxyethyl) carbamate and 6-benzylaminopurine. White solid, yield: 23.21%, melting point ：121.0～121.8℃,HPLC：98.42％.1H NMR(500MHz,DMSO-d6):δ9.60(d,J＝10.1Hz,1H),8.60(s,1H),8.48(s,1H),8.29(s,1H),7.98(s,1H),7.35(d,J＝7.4Hz,2H),7.29(t,J＝7.5Hz,2H),7.21(t,J＝7.2Hz,1H),6.92(d,J＝10.1Hz,1H),4.72(s,2H),4.56(t,J＝9.3Hz,3H),4.42(dd,J＝8.8,5.6Hz,1H),2.38(s,3H).13C NMR(500MHz,DMSO-d6):δ155.52,154.90,153.76,151.93,149.63,140.31,138.94,138.30,133.50,128.68,127.68,127.13,118.14,99.55,70.82,63.81,45.80,43.45,14.33.HRMS(ESI)m/z calcd for[C21H20Cl3N9O4+H]+:568.0704,found:568.0795.
Example 21
2- (2-Methyl-5-nitro-1H-imidazol-1-yl) -ethyl- (1- (6-amino-2-chloro-9H-purin-9-yl) -2, 2-trichloroethyl) -carbamic acid ester (8 e)
Steps 1 and 2 are the same as in example 1.
Compound 8e was obtained according to the synthesis procedure of example 1, step 3 starting from intermediate 2- (2-methyl-5-nitro-1H-imidazolin-1-yl) ethyl (2, 2-trichloro-1-hydroxyethyl) carbamate and 2-chloroadenine. White solid, yield: 20.18%, melting point ：196.5～197.6℃,HPLC：98.62％.1H NMR(500MHz,DMSO-d6):δ9.61(d,J＝10.0Hz,1H),8.45(s,1H),7.99(d,J＝20.3Hz,3H),6.73(d,J＝9.8Hz,1H),4.57(d,J＝9.3Hz,3H),4.44(d,J＝8.1Hz,1H),2.38(s,3H).13C NMR(500MHz,DMSO-d6):δ157.40,155.54,154.23,151.91,151.23,138.96,138.72,133.47,116.94,99.23,70.90,63.85,45.79,14.32.HRMS(ESI)m/z calcd for[C14H13Cl4N9O4+H]+:511.9845,found:511.9915.
Example 22
2- (2-Methyl-5-nitro-1H-imidazol-1-yl) -ethyl- (2, 2-trichloro-1- ((2, 6-diamino) -9H-purin-9-yl) -ethyl) -carbamic acid ester (8 f)
Steps 1 and 2 are the same as in example 1.
The procedure of example 1, step 3, was followed using the intermediates 2- (2-methyl-5-nitro-1H-imidazolin-1-yl) ethyl (2, 2-trichloro-1-hydroxyethyl) carbamate and 2, 6-diaminopurine as starting materials to give compound 8f. Pale yellow solid, yield: 25.36%, melting point ：165.8～166.6℃,HPLC：98.68％.1H NMR(400MHz,DMSO):δ9.47(d,J＝10.2Hz,1H),8.03(d,J＝7.8Hz,2H),6.85(s,2H),6.73(d,J＝10.1Hz,1H),6.07(s,2H),4.58(ddd,J＝16.2,9.9,5.1Hz,3H),4.39(dd,J＝7.6,4.6Hz,1H),2.40(s,3H).13C NMR(400MHz,DMSO):δ161.21,156.74,155.56,152.60,152.00,138.97,134.48,133.53,111.95,100.10,70.22,63.64,45.88,14.35.HRMS(ESI)m/z calcd for[C14H15Cl3N10O4+H]+:493.0343,found:493.0416.
Example 23
2- (2-Methyl-5-nitro-1H-imidazol-1-yl) -ethyl- (1- (6-amino-2-fluoro-9H-purin-9-yl) -2, 2-trichloroethyl) -carbamic acid ester (8 g)
Steps 1 and 2 are the same as in example 1.
The procedure of example 1, step 3, was followed using as starting materials the intermediate 2- (2-methyl-5-nitro-1H-imidazolin-1-yl) ethyl (2, 2-trichloro-1-hydroxyethyl) carbamate and 2-fluoroadenine to give 8g of compound. White solid, yield: 19.26%, melting point ：168.1～169.6℃,HPLC：96.92％.1H NMR(500MHz,DMSO-d6):δ9.60(d,J＝10.0Hz,1H),8.42(s,1H),8.00(d,J＝31.1Hz,3H),6.70(d,J＝10.0Hz,1H),4.57(d,J＝10.1Hz,3H),4.44(d,J＝7.5Hz,1H),2.38(s,3H).13C NMR(500MHz,DMSO-d6)δ160.25,158.62,158.40,158.23,155.54,151.91,151.71,151.55,138.96,138.61,133.46,116.28,99.26,71.02,63.85,45.79,14.30.HRMS(ESI)m/z calcd for[C14H13Cl3FN9O4+H]+:496.0140,found:496.0222.
Example 24
2- (2-Methyl-5-nitro-1H-imidazol-1-yl) -ethyl- (1- (2-amino-6-chloro-9H-purin-9-yl) -2, 2-trichloroethyl) -carbamic acid ester (8H)
Steps 1 and 2 are the same as in example 1.
The procedure of example 1, step 3, was followed using the intermediate 2- (2-methyl-5-nitro-1H-imidazolin-1-yl) ethyl (2, 2-trichloro-1-hydroxyethyl) carbamate and 2-amino-6-chloropurine as starting materials to give compound 8H. White solid, yield: 22.16%, melting point ：140.4～140.9℃,HPLC：97.69％.1H NMR(500MHz,DMSO-d6):δ9.61(d,J＝9.9Hz,1H),8.40(s,1H),7.96(s,1H),7.26(s,2H),6.78(d,J＝10.0Hz,1H),4.53(t,J＝35.8Hz,4H),2.38(s,3H).13C NMR(500MHz,DMSO-d6):δ160.75,155.58,154.62,151.90,150.53,139.79,139.01,133.45,122.32,99.33,70.70,63.74,45.83,14.33.HRMS(ESI)m/z calcd for[C14H13Cl4N9O4+H]+:511.9845,found:511.9923.
Example 25
2- (2-Methyl-5-nitro-1H-imidazol-1-yl) -ethyl- (2, 2-trichloro-1- (6-chloro-2-fluoro-9H-purin-9-yl) -ethyl) -carbamic acid ester (8 i)
Steps 1 and 2 are the same as in example 1.
The procedure of example 1, step 3, was followed using the intermediate 2- (2-methyl-5-nitro-1H-imidazolin-1-yl) ethyl (2, 2-trichloro-1-hydroxyethyl) carbamate and 2-fluoro-6-chloropurine as starting materials to give compound 8i. White solid, yield: 23.16%, melting point ：171.3～172.9℃,HPLC：98.85％.1H NMR(500MHz,DMSO-d6):δ9.80(d,J＝9.7Hz,1H),8.97(s,1H),7.92(s,1H),6.82(d,J＝10.0Hz,1H),4.60–4.47(m,4H),2.38(s,3H).13C NMR(500MHz,DMSO-d6):δ157.93,156.21,155.56,154.42,154.28,152.12,151.98,151.87,145.47,138.99,133.39,129.60,98.42,71.92,63.92,45.76,14.36.HRMS(ESI)m/z calcd for[C14H11Cl4N8O4+H]+:514.9641,found:514.9718.
Example 26
2- (2-Methyl-5-nitro-1H-imidazol-1-yl) -ethyl- (1- (2-amino-6-benzyloxy-9H-purin-9-yl) -2, 2-trichloroethyl) -carbamic acid ester (8 j)
Steps 1 and 2 are the same as in example 1.
The procedure of example 1, step 3, was followed using the intermediate 2- (2-methyl-5-nitro-1H-imidazolin-1-yl) ethyl (2, 2-trichloro-1-hydroxyethyl) carbamate and O-6-benzylguanine as starting materials to give compound 8j. White solid, yield: 24.53%, melting point ：171.6～172.8℃,HPLC：96.77％.1H NMR(500MHz,DMSO-d6):δ9.54(d,J＝10.1Hz,1H),8.17(s,1H),8.00(s,1H),7.52(d,J＝7.3Hz,2H),7.40(t,J＝7.3Hz,2H),7.38–7.34(m,1H),6.79(d,J＝7.3Hz,3H),5.50(s,2H),4.56(t,J＝10.3Hz,3H),4.41(d,J＝10.3Hz,1H),2.39(s,3H).13C NMR(500MHz,DMSO-d6):δ160.69,155.57,155.03,151.95,138.98,136.91,136.74,133.50,129.04,128.89,128.59,112.66,99.77,70.47,67.61,63.68,45.86,14.33.HRMS(ESI)m/z calcd for[C21H20Cl3N9O5+H]+:584.0653,found:584.0735.
Example 27
2- (2-Methyl-5-nitro-1H-imidazol-1-yl) -ethyl- (2, 2-trichloro-1- (2, 6-dichloro-9H-purin-9-yl) -ethyl) -carbamic acid ester (8 k)
Steps 1 and 2 are the same as in example 1.
The procedure of example 1, step 3, was followed using the intermediates 2- (2-methyl-5-nitro-1H-imidazolin-1-yl) ethyl (2, 2-trichloro-1-hydroxyethyl) carbamate and 2, 6-dichloropurine as starting materials to give compound 8k. White solid, yield: 19.33%, melting point ：129.9～131.7℃,HPLC：99.76％.1H NMR(500MHz,DMSO-d6):δ9.77(d,J＝9.8Hz,1H),9.01(d,J＝21.3Hz,1H),7.91(s,1H),6.86(d,J＝9.8Hz,1H),4.59–4.46(m,4H),2.37(s,3H).13C NMR(500MHz,DMSO-d6):δ155.56,153.83,152.55,151.87,151.22,145.42,139.01,133.38,130.10,98.46,71.80,63.92,45.76,14.37.HRMS(ESI)m/z calcd for[C14H11Cl5N8O4+H]+:530.9346,found:530.9421.
Example 28
2- (2-Methyl-5-nitro-1H-imidazol-1-yl) -ethyl- (1- (2-acetyl-amino-6-oxo-3, 6-dihydro-9H-purin-9-yl) -2, 2-trichloroethyl) -carbamic acid ester (8 l)
Steps 1 and 2 are the same as in example 1.
The procedure of example 1, step 3, was followed using the intermediate 2- (2-methyl-5-nitro-1H-imidazolin-1-yl) ethyl (2, 2-trichloro-1-hydroxyethyl) carbamate and N-2-acetylguanine as starting materials to give compound 8l. Pale yellow solid, yield: 22.16%, melting point ：197.5～198.3℃,HPLC：99.67％.1H NMR(400MHz,DMSO-d6):δ12.13(s,1H),11.90(s,1H),9.67(d,J＝9.1Hz,1H),8.29(s,1H),7.99(s,1H),6.76(d,J＝9.8Hz,1H),4.50(d,J＝54.2Hz,4H),2.38(s,3H),2.19(s,3H).13C NMR(500MHz,DMSO-d6):δ174.19,155.64,155.20,151.91,149.53,149.04,139.04,136.98,133.50,119.25,99.23,70.80,63.69,45.89,24.24,14.32.HRMS(ESI)m/z calcd for[C16H16Cl3N9O6+H]+:536.0289,found:536.0362.
Example 29
2- (2-Methyl-5-nitro-1H-imidazol-1-yl) -ethyl- (1- (6-amino-9H-purin-9-yl) -2, 2-trichloroethyl) -carbamic acid ester (8 m)
Steps 1 and 2 are the same as in example 1.
The procedure of example 1, step 3, was followed using the intermediate 2- (2-methyl-5-nitro-1H-imidazolin-1-yl) ethyl (2, 2-trichloro-1-hydroxyethyl) carbamate and adenine as starting materials to give compound 8m. White solid, yield: 12.36%, melting point ：187.1～187.9℃,HPLC：98.68％.1H NMR(400MHz,DMSO-d6):δ9.59(d,J＝10.1Hz,1H),8.46(s,1H),8.21(s,1H),7.98(s,1H),7.48(s,2H),6.89(d,J＝10.1Hz,1H),4.61-4.52(m,3H),4.47-4.39(m,1H),2.38(s,3H).13C NMR(101MHz,DMSO-d6):δ156.60,155.57,153.74,151.98,150.22,138.96,138.31,133.47,117.59,99.66,70.90,63.84,45.90,14.51.HRMS(ESI)m/z calcd for[C14H14Cl3N9O4+H]+:478.0234,found:478.0313.
Example 30
Benzyl- (1- (6-amino-2-chloro-9H-purin-9-yl) -2, 2-trichloroethyl) -carbamic acid ester (9 a)
The preparation was carried out in the same way as in example 1, step 2, starting from benzyl carbamate. Obtaining an intermediate (2, 2-trichloro-1-hydroxyethyl) benzyl carbamate crude product, and directly putting the crude product into the next reaction.
Compound 9a was obtained according to the synthesis method of example 1, step 3 starting from intermediate (benzyl 2, 2-trichloro-1-hydroxyethyl) carbamate and 2-chloroadenine. White solid, yield: 21.32%, melting point ：198.6～199.9℃,HPLC：98.49％.1H NMR(400MHz,DMSO-d6):δ9.69(d,J＝10.1Hz,1H),8.49(s,1H),8.05(s,2H),7.37(dd,J＝15.1,7.4Hz,5H),6.87(d,J＝10.2Hz,1H),5.21–5.12(m,2H).13C NMR(400MHz,DMSO-d6):δ157.42,155.85,154.31,151.25,138.78,136.18,128.95,128.86,128.80,116.92,99.48,70.91,67.70.HRMS(ESI)m/z calcd for[C15H12Cl4N6O2+H]+:448.9776,found:448.9853.
Example 31
(1-Methyl-5-nitro-1H-imidazol-2-yl) -methyl- (1- (6-amino-2-chloro-9H-purin-9-yl) -2, 2-trichloroethyl) -carbamic acid ester (9 b)
The preparation was carried out in the same way as in example 1, step 2, starting from lonidazole. Obtaining an intermediate (1-methyl-5-nitro-1H-imidazoline-2-yl) methyl (2, 2-trichloro-1-hydroxyethyl) carbamate crude product, and directly putting the crude product into the next reaction.
Compound 9b was obtained according to the synthesis method of example 1, step 3 starting from the intermediate (1-methyl-5-nitro-1H-imidazolin-2-yl) methyl (2, 2-trichloro-1-hydroxyethyl) carbamate and 2-chloroadenine. White solid, yield: 23.16%, melting point ：185.8～186.3℃,HPLC：98.24％.1H NMR(500MHz,DMSO-d6):δ9.87(d,J＝10.0Hz,1H),8.48(s,1H),8.06(d,J＝12.8Hz,3H),6.83(d,J＝10.0Hz,1H),5.32(q,J＝13.6Hz,2H),3.91(s,3H).13C NMR(500MHz,DMSO-d6):δ157.42,155.27,154.30,151.26,147.57,139.93,138.75,132.24,116.90,99.37,70.93,59.34,33.98.HRMS(ESI)m/z calcd for[C13H11Cl4N9O4+H]+:497.9688,found:497.9766.
Example 32
Ethyl- (1- (6-amino-2-chloro-9H-purin-9-yl) -2, 2-trichloroethyl) -carbamic acid ester (9 c)
The preparation was carried out in the same manner as in step 2 of example 1, starting from urethane. Obtaining an intermediate (2, 2-trichloro-1-hydroxyethyl) ethyl carbamate crude product, and directly putting the crude product into the next reaction.
Compound 9c was obtained according to the synthesis method of example 1, step 3 starting from the intermediate (2, 2-trichloro-1-hydroxyethyl) carbamate and 2-chloroadenine. White solid, yield: 19.89%, melting point ：219.5～220.4℃,HPLC：98.67％.1H NMR(400MHz,DMSO-d6):δ9.53(d,J＝10.2Hz,1H),8.51(s,1H),8.04(s,2H),6.83(d,J＝10.2Hz,1H),4.14(ddd,J＝10.7,7.0,3.7Hz,2H),1.22(t,J＝7.1Hz,3H).13C NMR(400MHz,DMSO-d6):δ157.42,155.92,154.28,151.25,138.86,116.91,99.57,70.85,62.25,14.77.HRMS(ESI)m/z calcd for[C10H10Cl4N6O2+H]+:386.9616,found:386.96970.
Example 33
(1-Methyl-5-nitro-1H-imidazol-2-yl) -methyl- (1- (6-amino-2-fluoro-9H-purin-9-yl) -2, 2-trichloroethyl) -carbamic acid ester (9 d)
The preparation was carried out in the same way as in example 1, step 2, starting from lonidazole. Obtaining an intermediate (1-methyl-5-nitro-1H-imidazoline-2-yl) methyl (2, 2-trichloro-1-hydroxyethyl) carbamate crude product, and directly putting the crude product into the next reaction.
Compound 9d was obtained according to the synthesis method of example 1, step 3 starting from intermediate (1-methyl-5-nitro-1H-imidazolin-2-yl) methyl (2, 2-trichloro-1-hydroxyethyl) carbamate and 2-fluoroadenine. White solid, yield: 21.63%, melting point ：205.8～206.9℃,HPLC：99.00％.1H NMR(500MHz,DMSO-d6):δ9.85(d,J＝9.9Hz,1H),8.45(s,1H),8.04(d,J＝24.3Hz,3H),6.80(d,J＝9.9Hz,1H),5.33(q,J＝13.6Hz,2H),3.92(s,3H).13C NMR(500MHz,DMSO-d6):δ160.29,158.66,158.41,158.24,155.26,151.73,151.57,147.58,139.91,138.62,132.22,116.24,102.69,99.37,84.10,71.03,59.33,33.96.HRMS(ESI)m/z calcd for[C13H11Cl4N9O4+H]+:481.9984,found:482.0055.
Example 34
Ethyl- (1- (6-amino-2-fluoro-9H-purin-9-yl) -2, 2-trichloroethyl) -carbamic acid ester (9 e)
The preparation was carried out in the same manner as in step 2 of example 1, starting from urethane. Obtaining an intermediate (2, 2-trichloro-1-hydroxyethyl) ethyl carbamate crude product, and directly putting the crude product into the next reaction.
Compound 9e was obtained according to the synthesis method of example 1, step 3 starting from the intermediate (2, 2-trichloro-1-hydroxyethyl) urethane and 2-fluoroadenine. White solid, yield: 18.21%, melting point ：208.3～209.8℃,HPLC：99.12％.1H NMR(400MHz,DMSO-d6):δ9.50(d,J＝10.1Hz,1H),8.48(s,1H),8.05(d,J＝45.7Hz,2H),6.80(d,J＝10.2Hz,1H),4.14(dd,J＝13.3,6.4Hz,2H),1.21(t,J＝7.0Hz,3H).13C NMR(400MHz,DMSO-d6):δ160.29,158.65,158.41,158.24,155.91,151.73,151.57,138.73,116.29,116.26,99.60,70.96,62.21,14.75.HRMS(ESI)m/z calcd for[C10H10Cl4N6O2+H]+:370.9915,found:370.9988.
Example 35
2-N-morpholinoethyl- (1- (6-amino-2-fluoro-9H-purin-9-yl) -2, 2-trichloroethyl) -carbamic acid ester (9 f)
The preparation method is the same as in step 2 of example 1 using hydroxyethylmorpholine as starting material. The intermediate 2-morpholinoethyl- (2, 2-trichloro-1-hydroxyethyl) carbamate crude product is obtained and then directly put into the next reaction.
Compound 9f was obtained according to the synthesis method of example 1, step 3 starting from the intermediates 2-N-morpholinoethyl- (2, 2-trichloro-1-hydroxyethyl) carbamate and 2-fluoroadenine. Pale yellow solid, yield ：10.42％,HPLC：96.96％.1H NMR(500MHz,DMSO-d6):δ9.68(d,J＝9.1Hz,1H),8.55(s,1H),8.03(d,J＝38.0Hz,2H),6.79(d,J＝10.0Hz,1H),4.21(s,2H),3.03(d,J＝7.3Hz,4H),2.60(s,2H),2.46(s,4H).13C NMR(126MHz,DMSO-d6):δ158.32(d,J＝5.9Hz),155.78(s),138.96(s),130.10(s),102.96(s),99.51(s),83.94(s),71.03(s),70.20(s),65.83(s),65.54(s),56.68(d,J＝5.6Hz),53.32(s).HRMS(ESI)m/z calcd for[C14H18Cl3FN7O3+H]+:456.0515,found:456.0517.
Example 36
2-N-morpholinoethyl- (2, 2-trichloro-1- (2, 6-diamino-9H-purin-9-yl) -ethyl) -carbamic acid ester (9 g)
The preparation method is the same as in step 2 of example 1 using hydroxyethylmorpholine as starting material. The intermediate 2-morpholinoethyl- (2, 2-trichloro-1-hydroxyethyl) carbamate crude product is obtained and then directly put into the next reaction.
The synthesis of step 3 was followed by example 1 starting from the intermediate 2-morpholinoethyl- (2, 2-trichloro-1-hydroxyethyl) carbamate and 2-aminoadenine to give 9g of compound. White solid, yield ：10.07％,HPLC：97.8％.1H NMR(500MHz,DMSO-d6):δ9.44(d,J＝10.1Hz,1H),8.11(s,1H),6.90-6.80(m,3H),6.08(s,2H),4.18(s,2H),3.51(s,4H),2.51(s,2H),2.38(s,4H).NMR HRMS(ESI)m/z calcd for[C14H19Cl3N8O3+H]+:453.0718,found:453.0724.
Example 37
2- (Pyridin-2-yl) -ethyl- (1- (6-amino-2-fluoro-9H-purin-9-yl) -2, 2-trichloroethyl) -carbamic acid ester (9H)
The preparation method is the same as in example step 2, using 2-hydroxyethyl pyridine as starting material. The intermediate 2- (pyridine-2-yl) ethyl- (2, 2-trichloro-1-hydroxyethyl) carbamate crude product is obtained and then directly put into the next reaction.
The procedure of example 1, step 3, was followed using the intermediates 2- (pyridin-2-yl) ethyl- (2, 2-trichloro-1-hydroxyethyl) carbamate and 2-fluoroadenine as starting materials to give compound 9h. White solid, yield ：26.58％,HPLC：97.28％.1H NMR(500MHz,DMSO-d6)δ9.78(d,J＝9.7Hz,1H),8.75(s,1H),8.55(s,1H),8.37(s,1H),8.07–7.92(m,3H),7.81(s,1H),6.70(d,J＝9.9Hz,1H),4.51(d,J＝6.9Hz,2H),3.39(s,2H).13C NMR(126MHz,DMSO-d6)δ160.22(s),158.58(s),158.33(s),155.69(s),154.21(s),142.99(s),139.02(d,J＝3.0Hz),127.59(s),125.26(s),116.20(s),99.37(s),70.98(s),64.02(s),33.43(s).HRMS(ESI)m/z calcd for[C15H13Cl3FN7O2+H]+:448.0253,found:448.0257.
Example 38
2- (Pyridin-2-yl) -ethyl- (2, 2-trichloro-1- (2, 6-diamino-9H-purin-9-yl) -ethyl) -carbamic acid ester (9 i)
The preparation method is the same as in example step 2, using 2-hydroxyethyl pyridine as starting material. The intermediate 2- (pyridine-2-yl) ethyl- (2, 2-trichloro-1-hydroxyethyl) carbamate crude product is obtained and then directly put into the next reaction.
Compound 9i was obtained according to the synthesis method of example 1, step 3 starting from the intermediates 2- (pyridin-2-yl) ethyl- (2, 2-trichloro-1-hydroxyethyl) carbamate and 2-aminoadenine. White solid, yield ：27.38％,HPLC：95.32％.1H NMR(500MHz,DMSO)δ9.34(d,J＝10.1Hz,1H),8.48(d,J＝3.9Hz,1H),8.07(s,1H),7.69(t,J＝7.1Hz,1H),7.29(d,J＝7.6Hz,1H),7.24–7.19(m,1H),6.91–6.77(m,3H),6.08(s,2H),4.52–4.43(m,2H),3.08(t,J＝6.5Hz,2H).13C NMR(101MHz,DMSO)δ161.23(s),158.02(s),156.75(s),152.59(s),149.56(s),137.00(s),134.68(s),123.90(s),122.23(s),111.95(s),100.45(s),70.19(s),64.90(s),60.23(s),37.14(s).HRMS(ESI)m/z calcd for[C15H15Cl3N8O2+H]+:445.0456,found:445.0460.
Example 39
Benzyl- (1- (6-amino-2-fluoro-9H-purin-9-yl) -2, 2-trichloroethyl) -carbamic acid ester (9 j)
The preparation was carried out in the same way as in example 1, step 2, starting from benzyl carbamate. Obtaining an intermediate (2, 2-trichloro-1-hydroxyethyl) benzyl carbamate crude product, and directly putting the crude product into the next reaction.
Compound 9j was obtained according to the synthesis method of example 1, step 3 starting from intermediate (benzyl 2, 2-trichloro-1-hydroxyethyl) carbamate and 2-fluoroadenine. White solid, yield: 19.82%, melting point ：234.8～235.7℃,HPLC：96.24％.1H NMR(500MHz,DMSO-d6):δ9.67(d,J＝10.1Hz,1H),8.46(s,1H),8.06(d,J＝47.0Hz,2H),7.46-7.31(m,5H),6.82(d,J＝10.2Hz,1H),5.16(q,J＝12.2Hz,2H).13C NMR(126MHz,DMSO-d6):δ160.30,158.67,158.42,158.25,155.85,151.72,151.56,138.65,136.20,128.94,128.82,116.29,99.53,71.01,67.68.HRMS(ESI)m/z calcd for[C15H12Cl4N6O2+H]+:433.0071,found:433.0146.
Example 40
2-N-morpholinoethyl- (1- (4-amino-1H-pyrazolo [3,4-d ] pyrimidin-1-yl) -2, 2-trichloroethyl) -carbamic acid ester (10 a)
The preparation method is the same as in the step 2 of the example by taking hydroxyethyl morpholine as a starting material. The intermediate 2-morpholinoethyl- (2, 2-trichloro-1-hydroxyethyl) carbamate crude product is obtained and then directly put into the next reaction.
The procedure of example 1, step 3, was followed using as starting materials the intermediates 2-morpholinoethyl- (2, 2-trichloro-1-hydroxyethyl) carbamate and 4-aminopyrazolo [3,4-d ] pyrimidine to give compound 10a. White solid, yield ：19.56％,HPLC：95.17％.1H NMR(500MHz,DMSO)δ9.27(d,J＝7.9Hz,1H),8.38(s,1H),8.22(d,J＝23.0Hz,2H),7.79(s,1H),7.15(d,J＝9.7Hz,1H),4.14(s,2H),3.48(s,4H),2.51(s,2H),2.37(s,4H).13C NMR(126MHz,DMSO)δ158.49(s),157.03(s),155.34(s),135.09(s),99.91(s),99.65(s),71.72(s),66.48(s),62.90(s),57.14(s),53.79(s).HRMS(ESI)m/z calcd for[C14H18Cl3N7O3+H]+:438.0604,found:438.0610.
Example 41
2- (Pyridin-1-yl) -ethyl- (2, 2-trichloro-1- (4- (2, 2-trichloro-1- ((2- (2-pyridin-2-yl) -ethoxycarbonylamino-) -ethyl) -amino) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl-ethyl) -carbamate (10 b)
The preparation method is the same as in example step 2, using 2-hydroxyethyl pyridine as starting material. The intermediate 2- (pyridine-2-yl) ethyl- (2, 2-trichloro-1-hydroxyethyl) carbamate crude product is obtained and then directly put into the next reaction.
Compound 10b was obtained according to the synthesis procedure of example 1, step 3 starting from intermediate 2- (pyridin-2-yl) ethyl- (2, 2-trichloro-1-hydroxyethyl) carbamate and 4-aminopyrazolo [3,4-d ] pyrimidine. White solid, yield ：9.84％,HPLC：97.73％.1H NMR(500MHz,DMSO)δ9.34(d,J＝9.2Hz,1H),9.06(d,J＝7.8Hz,1H),8.61(s,1H),8.51(s,1H),8.48–8.43(m,2H),8.39–8.31(m,1H),7.66(s,2H),7.29(d,J＝7.3Hz,2H),7.19(d,J＝4.4Hz,4H),4.42(dd,J＝8.0,5.7Hz,4H),3.04(d,J＝7.7Hz,4H).13C NMR(126MHz,DMSO)δ158.09(d,J＝12.8Hz),156.30(s),155.89(s),155.31(s),149.51(d,J＝5.0Hz),136.89(s),134.91(s),123.91(d,J＝6.1Hz),122.14(s),101.65(s),100.39(s),99.39(s),71.92(s),68.88(s),64.88(s),64.50(s),37.27(d,J＝11.4Hz).HRMS(ESI)m/z calcd for[C25H23Cl6N9O4+H]+:724.0077,found:724.0083.
Example 42
2- (Pyridin-2-yl) -ethyl- (2, 2-trichloro-1- (4-hydroxy-1H-pyrazolo [3,4-d ] pyrimidin-1-yl) -ethyl) -carbamic acid ester (10 c)
The preparation method is the same as in example step 2, using 2-hydroxyethyl pyridine as starting material. The intermediate 2- (pyridine-2-yl) ethyl- (2, 2-trichloro-1-hydroxyethyl) carbamate crude product is obtained and then directly put into the next reaction.
The procedure of example 1, step 3, was followed using the intermediates 2- (pyridin-2-yl) ethyl- (2, 2-trichloro-1-hydroxyethyl) carbamate and 4-hydroxypyrazolo [3,4-d ] pyrimidine as starting materials to give compound 10c. Pale yellow solid, yield ：7.22％,HPLC：95.06％.1H NMR(500MHz,DMSO)δ12.49(s,1H),9.41(d,J＝9.6Hz,1H),8.46(s,1H),8.28(s,1H),8.22(s,1H),7.68(t,J＝6.7Hz,1H),7.31(d,J＝7.0Hz,1H),7.25–7.19(m,1H),7.12(d,J＝9.8Hz,1H),4.42(dd,J＝13.4,6.5Hz,2H),3.05(t,J＝6.3Hz,2H).13C NMR(101MHz,DMSO)δ157.99(s),157.44(s),154.15(s),149.73(s),149.45(s),137.04(s),136.81(s),124.05(s),122.17(s),116.17(s),106.07(s),99.15(s),72.11(s),64.92(s),37.14(s)HRMS(ESI)m/z calculated for[C15H13Cl3N6O3+H]+:431.0187,found:431.0193
Example 43
2- (Pyridin-1-yl) -ethyl- (2, 2-trichloro-1- (4- (2, 2-trichloro-1- (2- (2-pyridin-2-yl) -ethoxycarbonylamino-) -ethoxy) -1H-pyrazolo [3,4-d ] pyrimidin-1-yl-ethyl) -carbamate (10 d)
The preparation method is the same as in example step 2, using 2-hydroxyethyl pyridine as starting material. The intermediate 2- (pyridine-2-yl) ethyl- (2, 2-trichloro-1-hydroxyethyl) carbamate crude product is obtained and then directly put into the next reaction.
The procedure of example 1, step 3, was followed using the intermediates 2- (pyridin-2-yl) ethyl- (2, 2-trichloro-1-hydroxyethyl) carbamate and 4-hydroxypyrazolo [3,4-d ] pyrimidine as starting materials to give compound 10d. Pale yellow solid, yield ：7.22％,HPLC：96.23％.1H NMR(500MHz,DMSO)δ9.48(s,1H),9.36(d,J＝9.4Hz,1H),9.14(s,1H),8.65(s,1H),8.46(s,2H),7.68(s,2H),7.39(d,J＝10.0Hz,1H),7.30(d,J＝6.7Hz,2H),7.20(s,2H),6.91(d,J＝14.1Hz,1H),4.48(dd,J＝15.2,7.7Hz,4H),3.15–3.01(m,4H).
Example 44
2- (2-Methyl-5-nitro-1H-imidazol-1-yl) -ethyl- (1- (6-amino-1H-pyrazolo [3,4-d ] pyrimidin-1-yl) -2, 2-trichloroethyl) -carbamic acid ester (10 e)
Steps 1 and 2 are the same as in example 1.
The procedure of example 1, step 3, was followed using the intermediates 2- (2-methyl-5-nitro-1H-imidazolin-1-yl) ethyl (2, 2-trichloro-1-hydroxyethyl) carbamate and 6-aminopyrazolo [3,4-d ] pyrimidine as starting materials to give compound 10e. Pale yellow solid, yield ：14.65％,HPLC：98.62％.1H NMR(500MHz,DMSO)δ9.63(d,J＝10.9Hz,1H),8.53(s,1H),8.05(d,J＝47.0Hz,3H),7.73(s,1H),6.81(d,J＝9.8Hz,1H),4.19(d,J＝5.7Hz,4H),2.37(s,3H).HRMS(ESI)m/z calcd for[C14H14Cl3N9O4+H]+:478.0302,found:478.0309.
Example 45
2- (2-Methyl-5-nitro-1H-imidazol-1-yl) -ethyl- (2, 2-trichloro-1- (6-chloro-1H-pyrazolo [3,4-d ] pyrimidin-1-yl) -ethyl) -carbamic acid ester (10 f)
Steps 1 and 2 are the same as in example 1.
The procedure of example 1, step 3, was followed using the intermediates 2- (2-methyl-5-nitro-1H-imidazolin-1-yl) ethyl (2, 2-trichloro-1-hydroxyethyl) carbamate and 6-chloropyrazolo [3,4-d ] pyrimidine as starting materials to give the hydrochloride salt of compound 10 f. Pale yellow solid, yield ：9.08％,HPLC：95.06％.1H NMR(500MHz,DMSO)δ11.89(d,J＝3.4Hz,1H),9.64(d,J＝9.5Hz,1H),8.98(s,1H),8.01(d,J＝3.7Hz,1H),7.99(s,1H),6.89(d,J＝10.0Hz,1H),4.54(s,4H),2.40(s,3H).HRMS(ESI)m/z calcd for[C14H12Cl4N8O4+H]+:496.9803,found:497.0073.
Example 46
Cancer cell proliferation inhibition experiment
To verify the anticancer activity of the designed synthetic compounds, two major activity tests were designed for this subject. Firstly, screening proliferation inhibition activity of breast cancer MCF-7, melanoma A375, lung adenocarcinoma A549, hepatocellular carcinoma HepG2, cervical carcinoma Hela and ovarian cancer Carvo-3,6 cell lines by using an MTT method, wherein the positive control drug is a specific inhibitor Apcin of Cdc 20.
The principle of the MTT method is that succinate dehydrogenase in mitochondria of living cells can reduce exogenous tetrazolium bromide (MTT) into insoluble blue-violet crystals and deposit in cells, whereas dead cells do not. DMSO can dissolve purple crystals in cells, and the light absorption value can be detected by an enzyme-linked immunosorbent assay (ELISA) at the wavelength of 490nm, so that the number of living cells can be reflected.
The final IC 50 values were obtained by three parallel experiments using the MTT method described above.
(1) Experimental materials
A. cell lines: MCF-7, A375, A549, hepG2, hela, carvo-3 cell lines
B. Reagents and instrumentation: tetramethyl azoazole salt (MTT) and dimethyl sulfoxide (DMSO) were purchased from Sigma-Aldrich company; neonatal bovine serum, fetal bovine serum, pancreatin, RPMI-1640, DMEM, and penicillin-streptomycin were purchased from Gibco; other reagents were purchased from Sigma-Aldrich company without specific description; CO 2 incubator (Thermo), microplate reader (Thermo) and fluorescent inverted microscope (Olympus).
(2) Experimental details
A. The experiment (MTT method) was performed by taking cells with a proportion of viable cells of 90% or more. The cells in log phase are collected, the concentration of the cell suspension is adjusted to 4-5 multiplied by 10 4/mL, 100 mu L of each hole is added, and the cells to be tested are plated to adjust the density to 4000-5000/hole.
Incubating at 37 ℃ with 5% CO 2% until cell monolayer is fully paved at the bottom of the well (96-well flat bottom plate), adding medicines with different concentration gradients, so that the final concentration of the compounds in the well is 300, 100, 30, 10, 3, 1, 0.3, 0.1, 0.03 and 0 mu M/mL, and making 3 compound wells for comparison of group-to-group differences.
C. placing into a constant temperature incubator with 5% CO 2 and 37 ℃ for 48 hours, and observing under an inverted microscope.
D. mu.L of MTT solution (5 mg/mL, i.e., 0.5% MTT) was added to each well and the incubation was continued for 4 hours.
E, stopping culturing after 4 hours, and carefully sucking out the culture solution in the hole.
F. 150 μl DMSO was added to each well, and the mixture was shaken on a shaker for 10min at low speed to allow the crystals to dissolve well. The absorbance of each well was measured at 490nm wavelength by ELISA and the inhibition was calculated.
G. at the same time, zeroing wells (medium, MTT, DMSO) and control wells (cells, drug dissolution medium of the same concentration, culture broth, MTT, DMSO) were set.
(3) Data processing
Three replicates were repeated for each concentration gradient, and compound inhibition (IC 50 values) was calculated using SPSS13.0 software, fitting curves with concentrations on the abscissa and inhibition on the ordinate. The absorbance OD value of each well is measured at the detection wavelength of 490nm on an enzyme-labeled instrument, and the cell growth inhibition rate is calculated according to the following formula:
Table 1: inhibition data of test compounds on cancer cell lines
The results show that:
compared with the specific Cdc20 inhibitor Apcin, the anticancer activity of most of the obtained target compounds is improved. Compounds containing dichloro-substituted pyrimidine (7 a-7 d) and 2-fluoroadenine side chains (8 g, 9d, 9e, 9f, 9 h) exhibited the most potent antiproliferative activity. To further confirm the targeting of the compounds, the cell cycle and apoptosis studies designed in this invention were continued with the selection of 7d and 9f with the best activity.
Example 47
Human hepatoma cell (Hep-G2) cycle experiment
(1) Experimental materials
A. Cell lines: human hepatoma cell HepG2
B. Reagents and instrumentation: multifunctional imaging systems are available from Biotek corporation; phosphorylated histone H3 antibody was purchased from CST; the manufacturers of PBS, pancreatin digest, DMSO, fetal bovine serum, DMEM, 96-well cell culture plates, T25 cell culture flasks, CO2 cell culture boxes, low-speed centrifuges, cell counters, and biological clean benches were the same as in the cancer cell proliferation inhibition experiments.
(2) Experimental details
A. Taking HepG2 cells growing in the logarithmic phase, washing with 1 XPBS for 3 times each for 2min; pancreatin digestion was added to the flask/petri dish and digested in a CO 2 incubator for 2min. Adding a culture medium (DMEM) into the culture flask/culture dish to stop digestion, and blowing the cells with a sterilization straw until the cells are in a single suspension state;
b. count with a cell counting plate and plate.
C. And (5) feeding medicines, and incubating for 2d in an incubator.
D. the medium was aspirated from each well, 100. Mu.l of 4% paraformaldehyde was added to each well, and incubated at room temperature for 15min.
E. Aspirate and rinse three times with PBS.
F. 0.1ml of permeate was added to each well and incubated at room temperature for 15min.
G. The permeate was aspirated and washed three times with PBS.
H. 100 μl of blocking buffer was added to each well and incubated for 15min at room temperature.
I. the blocking buffer was aspirated, 50 μl of primary antibody solution was added to each well and incubated overnight at 4deg.C.
J. The primary antibody solution was aspirated and washed three times with PBS, 50. Mu.l of secondary antibody solution was added to each well and incubated at room temperature for 60min (protected from light).
K. The secondary antibody solution was aspirated and washed three times with PBS, 100. Mu.l PBS was added to each well, and imaging was performed.
The immunofluorescence assay results of drug-treated HepG2 cell lines are shown in fig. 4. Drug group: compound Apcin (100 μm), 7d (30 μm), 9f (0.3 μm). Drug treatment time: 48h.
The 30. Mu.M compound 7d and 0.3. Mu.M compound 9f induced an increase in phosphohistone H3 positive cells, i.e., from 3.8% to 6.9% and 10.0%, respectively, compared to the solvent control group in HepG2 cells. Whereas 100. Mu.M positive compound Apcin resulted in an approximately 2.2-fold increase in PHH3 positive cell number compared to the DMSO control. Compounds 7d and 9f showed greater potency in blocking cell mitotic exit than the positive control drug Apcin, while compound 9f was the most potent mitotic blocker, which showed a strong M-phase blocking effect at 0.3 μm.
Example 48
Apoptosis experiment of human liver cancer cell (Hep-G2)
(1) Experimental materials
A. Cell lines: human hepatoma cell HepG2
B. Reagents and instrumentation: multifunctional imaging systems are available from Biotek corporation; the Annexin-FITC/PI apoptosis kit is purchased from Soy Bao company; the manufacturers of PBS, pancreatin digest, DMSO, fetal bovine serum, RPMI 1640, 24-well cell culture plates, T25 cell culture flasks, CO2 cell culture boxes, low-speed centrifuges, cell counters, and biological clean benches were the same as in the cancer cell proliferation inhibition experiments.
(2) Experimental procedure
A. HepG2 cells grown in log phase were taken, washed 3 times with 1 XPBS for 2min each, and digested with pancreatin in a flask/petri dish and 2min in a CO 2 incubator. The digestion was terminated by adding a medium (DMEM) to the flask/petri dish and the cells were blown into individual suspension with a sterile pipette. Counts were performed with a cell counting plate and plated.
B. Dosing and incubation overnight at 37 ℃.
C. 3ml Binding Buffer (10X) to 30ml of deionized water was diluted with 27ml of deionized water, 3ml each time.
D. Mu.l ANNEXIN V-FITC was added and stained for 10 minutes, at room temperature, protected from light, and the mixture was shaken by hand or several times.
E. mu.l PI was added, incubated at room temperature in the dark for 5min.
F. the staining solution was aspirated, rinsed with PBS and used for imaging.
The results of apoptosis experiments of drug-treated HepG2 cell lines are shown in fig. 5. Drug group: compound Apcin (100 μm), 9f (0.3 μm); drug treatment time: 24h. When the drug was treated for 24 hours, apcin at high concentrations had a significant pro-apoptotic effect, with most cells in the early stage of apoptosis, but late apoptotic cells were also present. Whereas high concentrations of 9f have a significant pro-apoptotic effect and most cells are in the early stage of apoptosis. Compound 7d (30 μm) also had pro-apoptotic effects.
The above mechanism studies have preliminarily shown that compounds 9f and 7d can significantly inhibit proliferation of cancer cells, and the studies have shown that the compounds designed for this purpose exert the effect of inhibiting proliferation of cancer cells by blocking cell cycle and promoting apoptosis.
The foregoing examples are set forth in order to provide a more thorough description of the present application and are not intended to limit the scope of the application, and various modifications of the application, which are equivalent to those skilled in the art upon reading the present application, will fall within the scope of the application as defined in the appended claims.

Claims (2)

1. A (1, 1-trichloro-2) carbamate derivative, characterized by the following structural formula:
2. use of a (1, 1-trichloro-2) carbamate derivative according to claim 1 for the preparation of a medicament for the treatment of lung adenocarcinoma a549, hepatocellular carcinoma HepG2 and ovarian cancer Carvo-3.
CN202211387962.8A 2020-04-15 2020-05-07 (1, 1-Trichloro-2) carbamate derivative and preparation method and application thereof Active CN115611867B (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN202211387962.8A CN115611867B (en) 2020-04-15 2020-05-07 (1, 1-Trichloro-2) carbamate derivative and preparation method and application thereof

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
CN202010296082 2020-04-15
CN2020102960824 2020-04-15
CN202010377518.2A CN111606891B (en) 2020-04-15 2020-05-07 (1,1,1-trichloro-2) carbamate derivative and preparation method and application thereof
CN202211387962.8A CN115611867B (en) 2020-04-15 2020-05-07 (1, 1-Trichloro-2) carbamate derivative and preparation method and application thereof

Related Parent Applications (1)

Application Number Title Priority Date Filing Date
CN202010377518.2A Division CN111606891B (en) 2020-04-15 2020-05-07 (1,1,1-trichloro-2) carbamate derivative and preparation method and application thereof

Publications (2)

Publication Number Publication Date
CN115611867A CN115611867A (en) 2023-01-17
CN115611867B true CN115611867B (en) 2024-04-30

Family

ID=72203114

Family Applications (2)

Application Number Title Priority Date Filing Date
CN202211387962.8A Active CN115611867B (en) 2020-04-15 2020-05-07 (1, 1-Trichloro-2) carbamate derivative and preparation method and application thereof
CN202010377518.2A Active CN111606891B (en) 2020-04-15 2020-05-07 (1,1,1-trichloro-2) carbamate derivative and preparation method and application thereof

Family Applications After (1)

Application Number Title Priority Date Filing Date
CN202010377518.2A Active CN111606891B (en) 2020-04-15 2020-05-07 (1,1,1-trichloro-2) carbamate derivative and preparation method and application thereof

Country Status (1)

Country Link
CN (2) CN115611867B (en)

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2005025515A2 (en) * 2003-09-12 2005-03-24 California Institute Of Technology Proteasome pathway inhibitors and related methods

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2005025515A2 (en) * 2003-09-12 2005-03-24 California Institute Of Technology Proteasome pathway inhibitors and related methods

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
Synergistic blockade of mitotic exit by two chemical inhibitors of the APC/C;Katharine L. Sackton 等;《Nature》;646-649 *

Also Published As

Publication number Publication date
CN115611867A (en) 2023-01-17
CN111606891B (en) 2023-01-17
CN111606891A (en) 2020-09-01

Similar Documents

Publication Publication Date Title
CN109689641B (en) Crystal form and salt form of substituted 2-hydrogen-pyrazole derivative and preparation method thereof
CN111454327B (en) NAMPT protein degradation targeting chimeric and preparation method and application thereof
EP2949647A1 (en) Deuterated phenyl amino pyrimidine compound and pharmaceutical composition containing same
TWI531573B (en) Novel compounds of reverse-turn mimetics and use therefor
CN110563703B (en) Compound for inducing PARP-1 degradation based on CRBN ligand, preparation method and application
KR20190098266A (en) Kinase inhibitors, which are substituted condensed heteroaryl group compounds, and their applications
CN103588792A (en) Pyrazolopyrimidine or pyrimidinopyrimidine compounds, preparation method therefor, pharmaceutical compositions thereof and applications thereof
KR20210038921A (en) Sulfoximine compounds and pharmaceutical compositions as bromodomain protein inhibitors and their medical use
CN112047950B (en) Imidazo pyrazine derivative and synthetic method and application thereof
US20210101881A1 (en) Pyrimidine compound, preparation method thereof and medical use thereof
WO2020253875A1 (en) Salts of compound, crystalline forms thereof, preparation method therefor and application thereof
JP2018135268A (en) Novel heteroaryl amino-3-pyrazole derivative and pharmacologically acceptable salt thereof
WO2019100743A1 (en) Parp-1 and pi3k dual target inhibitor comprising benzofuran
CN115611867B (en) (1, 1-Trichloro-2) carbamate derivative and preparation method and application thereof
CN109384788B (en) Purine series derivative and preparation method and application thereof
CN109641909A (en) The mechanism target and its treatment use of rapamycin signal pathway inhibitor
CN114315837B (en) Crystalline forms of ERK inhibitors and methods of making the same
CN109734677A (en) The small molecule compound and its application of inhibition of histone lysine methyltransferase NSD2
CN114409636B (en) Quinazolinone compound or pharmaceutically acceptable salt thereof, and preparation method and application thereof
CN113461661B (en) 6- (pyridin-3-yl) quinazoline-4 (3H) -ketone derivative and preparation and application thereof
CN112174958B (en) Pyrido [2,3-d ] pyrimidine compound and preparation method and application thereof
TW201738227A (en) Fused ring compound and preparation method, application and intermediate compound thereof
CN115304605B (en) Oxetane derivatives with antitumor activity, and preparation method and application thereof
WO2021164789A1 (en) Crystal form of pyrazolopyrimidine compound and use thereof
CN114829344B (en) Pyrimidine derivatives and pharmaceutical compositions containing the same

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
GR01 Patent grant
GR01 Patent grant