CN115746017B - Thienopyrimidine compound and preparation method and application thereof - Google Patents
Thienopyrimidine compound and preparation method and application thereof Download PDFInfo
- Publication number
- CN115746017B CN115746017B CN202211520649.7A CN202211520649A CN115746017B CN 115746017 B CN115746017 B CN 115746017B CN 202211520649 A CN202211520649 A CN 202211520649A CN 115746017 B CN115746017 B CN 115746017B
- Authority
- CN
- China
- Prior art keywords
- methyl
- cancer
- compound
- thienopyrimidine compound
- pyrimidine
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- -1 Thienopyrimidine compound Chemical class 0.000 title claims abstract description 50
- 238000002360 preparation method Methods 0.000 title claims abstract description 18
- 206010006187 Breast cancer Diseases 0.000 claims abstract description 16
- 208000026310 Breast neoplasm Diseases 0.000 claims abstract description 16
- 206010008342 Cervix carcinoma Diseases 0.000 claims abstract description 16
- 206010009944 Colon cancer Diseases 0.000 claims abstract description 16
- 208000006105 Uterine Cervical Neoplasms Diseases 0.000 claims abstract description 16
- 201000010881 cervical cancer Diseases 0.000 claims abstract description 16
- 208000029742 colonic neoplasm Diseases 0.000 claims abstract description 16
- 206010058467 Lung neoplasm malignant Diseases 0.000 claims abstract description 9
- 201000005202 lung cancer Diseases 0.000 claims abstract description 9
- 208000020816 lung neoplasm Diseases 0.000 claims abstract description 9
- 201000007270 liver cancer Diseases 0.000 claims abstract description 8
- 208000014018 liver neoplasm Diseases 0.000 claims abstract description 8
- 239000012271 PD-L1 inhibitor Substances 0.000 claims description 25
- 229940121656 pd-l1 inhibitor Drugs 0.000 claims description 25
- 239000002246 antineoplastic agent Substances 0.000 claims description 11
- 229940041181 antineoplastic drug Drugs 0.000 claims description 11
- OYRRZWATULMEPF-UHFFFAOYSA-N pyrimidin-4-amine Chemical compound NC1=CC=NC=N1 OYRRZWATULMEPF-UHFFFAOYSA-N 0.000 claims description 11
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims description 10
- 206010028980 Neoplasm Diseases 0.000 claims description 10
- WYURNTSHIVDZCO-UHFFFAOYSA-N Tetrahydrofuran Chemical compound C1CCOC1 WYURNTSHIVDZCO-UHFFFAOYSA-N 0.000 claims description 10
- 238000000034 method Methods 0.000 claims description 10
- 239000002904 solvent Substances 0.000 claims description 10
- 238000006243 chemical reaction Methods 0.000 claims description 9
- 239000012043 crude product Substances 0.000 claims description 8
- INQOMBQAUSQDDS-UHFFFAOYSA-N iodomethane Chemical compound IC INQOMBQAUSQDDS-UHFFFAOYSA-N 0.000 claims description 8
- PGTSGPCXPIFQEL-UHFFFAOYSA-N 1-methylindol-5-amine Chemical compound NC1=CC=C2N(C)C=CC2=C1 PGTSGPCXPIFQEL-UHFFFAOYSA-N 0.000 claims description 6
- LDNWUEWFBZXERC-UHFFFAOYSA-N 4-chloro-2-methylthiophene Chemical compound CC1=CC(Cl)=CS1 LDNWUEWFBZXERC-UHFFFAOYSA-N 0.000 claims description 6
- CZPWVGJYEJSRLH-UHFFFAOYSA-N Pyrimidine Chemical compound C1=CN=CN=C1 CZPWVGJYEJSRLH-UHFFFAOYSA-N 0.000 claims description 6
- 229960000583 acetic acid Drugs 0.000 claims description 5
- 239000012362 glacial acetic acid Substances 0.000 claims description 5
- YLQBMQCUIZJEEH-UHFFFAOYSA-N tetrahydrofuran Natural products C=1C=COC=1 YLQBMQCUIZJEEH-UHFFFAOYSA-N 0.000 claims description 5
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical group CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 4
- KEAYESYHFKHZAL-UHFFFAOYSA-N Sodium Chemical compound [Na] KEAYESYHFKHZAL-UHFFFAOYSA-N 0.000 claims description 4
- 238000006555 catalytic reaction Methods 0.000 claims description 4
- 230000008569 process Effects 0.000 claims description 4
- 229910000104 sodium hydride Inorganic materials 0.000 claims description 4
- 239000012312 sodium hydride Substances 0.000 claims description 4
- 238000000967 suction filtration Methods 0.000 claims description 4
- 238000000605 extraction Methods 0.000 claims description 3
- 238000002156 mixing Methods 0.000 claims description 3
- QIZMFTNGJPBSBT-UHFFFAOYSA-N thieno[3,2-d]pyrimidin-4-amine Chemical compound NC1=NC=NC2=C1SC=C2 QIZMFTNGJPBSBT-UHFFFAOYSA-N 0.000 claims description 2
- 201000001441 melanoma Diseases 0.000 claims 2
- 201000010893 malignant breast melanoma Diseases 0.000 claims 1
- 230000000259 anti-tumor effect Effects 0.000 abstract description 23
- IAKHMKGGTNLKSZ-INIZCTEOSA-N (S)-colchicine Chemical compound C1([C@@H](NC(C)=O)CC2)=CC(=O)C(OC)=CC=C1C1=C2C=C(OC)C(OC)=C1OC IAKHMKGGTNLKSZ-INIZCTEOSA-N 0.000 abstract description 12
- 230000000694 effects Effects 0.000 abstract description 9
- 229960001338 colchicine Drugs 0.000 abstract description 6
- 238000000746 purification Methods 0.000 abstract description 2
- 238000000926 separation method Methods 0.000 abstract description 2
- 241000699670 Mus sp. Species 0.000 description 24
- 150000001875 compounds Chemical class 0.000 description 24
- 210000004027 cell Anatomy 0.000 description 21
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 9
- 230000005764 inhibitory process Effects 0.000 description 9
- 239000013641 positive control Substances 0.000 description 6
- RBNBDIMXFJYDLQ-UHFFFAOYSA-N thieno[3,2-d]pyrimidine Chemical class C1=NC=C2SC=CC2=N1 RBNBDIMXFJYDLQ-UHFFFAOYSA-N 0.000 description 6
- 239000003814 drug Substances 0.000 description 5
- 239000007787 solid Substances 0.000 description 5
- 210000004881 tumor cell Anatomy 0.000 description 5
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 4
- 238000005481 NMR spectroscopy Methods 0.000 description 4
- HEDRZPFGACZZDS-MICDWDOJSA-N Trichloro(2H)methane Chemical compound [2H]C(Cl)(Cl)Cl HEDRZPFGACZZDS-MICDWDOJSA-N 0.000 description 4
- 201000011510 cancer Diseases 0.000 description 4
- 230000035484 reaction time Effects 0.000 description 4
- 238000011160 research Methods 0.000 description 4
- 238000001228 spectrum Methods 0.000 description 4
- 230000004614 tumor growth Effects 0.000 description 4
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 3
- 238000004440 column chromatography Methods 0.000 description 3
- 229940079593 drug Drugs 0.000 description 3
- 239000011259 mixed solution Substances 0.000 description 3
- 239000012074 organic phase Substances 0.000 description 3
- 125000000714 pyrimidinyl group Chemical group 0.000 description 3
- 239000000243 solution Substances 0.000 description 3
- 229910052717 sulfur Inorganic materials 0.000 description 3
- 239000011593 sulfur Substances 0.000 description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 3
- KDCGOANMDULRCW-UHFFFAOYSA-N 7H-purine Chemical compound N1=CNC2=NC=NC2=C1 KDCGOANMDULRCW-UHFFFAOYSA-N 0.000 description 2
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 2
- 208000024172 Cardiovascular disease Diseases 0.000 description 2
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 2
- NINIDFKCEFEMDL-UHFFFAOYSA-N Sulfur Chemical compound [S] NINIDFKCEFEMDL-UHFFFAOYSA-N 0.000 description 2
- YTPLMLYBLZKORZ-UHFFFAOYSA-N Thiophene Chemical group C=1C=CSC=1 YTPLMLYBLZKORZ-UHFFFAOYSA-N 0.000 description 2
- 230000001093 anti-cancer Effects 0.000 description 2
- 238000003556 assay Methods 0.000 description 2
- 230000037396 body weight Effects 0.000 description 2
- 229910052799 carbon Inorganic materials 0.000 description 2
- 230000010261 cell growth Effects 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 201000010099 disease Diseases 0.000 description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
- 238000001035 drying Methods 0.000 description 2
- 150000002391 heterocyclic compounds Chemical class 0.000 description 2
- 229910052739 hydrogen Inorganic materials 0.000 description 2
- 239000001257 hydrogen Substances 0.000 description 2
- 230000005918 in vitro anti-tumor Effects 0.000 description 2
- 230000002401 inhibitory effect Effects 0.000 description 2
- 229910052757 nitrogen Inorganic materials 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 238000012216 screening Methods 0.000 description 2
- 230000002195 synergetic effect Effects 0.000 description 2
- 238000010998 test method Methods 0.000 description 2
- 238000001644 13C nuclear magnetic resonance spectroscopy Methods 0.000 description 1
- 238000005160 1H NMR spectroscopy Methods 0.000 description 1
- 208000005623 Carcinogenesis Diseases 0.000 description 1
- 206010059866 Drug resistance Diseases 0.000 description 1
- 238000002965 ELISA Methods 0.000 description 1
- 208000017701 Endocrine disease Diseases 0.000 description 1
- 206010061218 Inflammation Diseases 0.000 description 1
- 238000000134 MTT assay Methods 0.000 description 1
- 231100000002 MTT assay Toxicity 0.000 description 1
- 208000012902 Nervous system disease Diseases 0.000 description 1
- 208000025966 Neurological disease Diseases 0.000 description 1
- 102000003993 Phosphatidylinositol 3-kinases Human genes 0.000 description 1
- 108090000430 Phosphatidylinositol 3-kinases Proteins 0.000 description 1
- PMZURENOXWZQFD-UHFFFAOYSA-L Sodium Sulfate Chemical compound [Na+].[Na+].[O-]S([O-])(=O)=O PMZURENOXWZQFD-UHFFFAOYSA-L 0.000 description 1
- 241001061127 Thione Species 0.000 description 1
- 208000036142 Viral infection Diseases 0.000 description 1
- 230000002159 abnormal effect Effects 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
- 230000029936 alkylation Effects 0.000 description 1
- 238000005804 alkylation reaction Methods 0.000 description 1
- 230000033115 angiogenesis Effects 0.000 description 1
- 230000000844 anti-bacterial effect Effects 0.000 description 1
- 230000000078 anti-malarial effect Effects 0.000 description 1
- 230000000840 anti-viral effect Effects 0.000 description 1
- 239000003430 antimalarial agent Substances 0.000 description 1
- 239000003963 antioxidant agent Substances 0.000 description 1
- 230000003078 antioxidant effect Effects 0.000 description 1
- 230000006907 apoptotic process Effects 0.000 description 1
- 230000006399 behavior Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- DZBUGLKDJFMEHC-UHFFFAOYSA-N benzoquinolinylidene Natural products C1=CC=CC2=CC3=CC=CC=C3N=C21 DZBUGLKDJFMEHC-UHFFFAOYSA-N 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 230000036952 cancer formation Effects 0.000 description 1
- 229910002091 carbon monoxide Inorganic materials 0.000 description 1
- 231100000504 carcinogenesis Toxicity 0.000 description 1
- 239000003054 catalyst Substances 0.000 description 1
- 230000003915 cell function Effects 0.000 description 1
- 239000006285 cell suspension Substances 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 239000013078 crystal Substances 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 208000037765 diseases and disorders Diseases 0.000 description 1
- 239000012039 electrophile Chemical class 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 230000012010 growth Effects 0.000 description 1
- 125000005843 halogen group Chemical group 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 206010073071 hepatocellular carcinoma Diseases 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-M hydroxide Chemical compound [OH-] XLYOFNOQVPJJNP-UHFFFAOYSA-M 0.000 description 1
- 208000026278 immune system disease Diseases 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 230000005917 in vivo anti-tumor Effects 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 230000006698 induction Effects 0.000 description 1
- 230000004054 inflammatory process Effects 0.000 description 1
- 239000003112 inhibitor Substances 0.000 description 1
- 231100000053 low toxicity Toxicity 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 208000030159 metabolic disease Diseases 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 238000000655 nuclear magnetic resonance spectrum Methods 0.000 description 1
- 230000000144 pharmacologic effect Effects 0.000 description 1
- 239000012071 phase Substances 0.000 description 1
- 230000019491 signal transduction Effects 0.000 description 1
- 239000012265 solid product Substances 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 238000004659 sterilization and disinfection Methods 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 230000002194 synthesizing effect Effects 0.000 description 1
- 238000010189 synthetic method Methods 0.000 description 1
- 125000003396 thiol group Chemical group [H]S* 0.000 description 1
- GLQWRXYOTXRDNH-UHFFFAOYSA-N thiophen-2-amine Chemical class NC1=CC=CS1 GLQWRXYOTXRDNH-UHFFFAOYSA-N 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 229910052723 transition metal Inorganic materials 0.000 description 1
- 150000003624 transition metals Chemical class 0.000 description 1
- 230000009385 viral infection Effects 0.000 description 1
- 238000009333 weeding Methods 0.000 description 1
- 230000004580 weight loss Effects 0.000 description 1
Abstract
The invention discloses a thienopyrimidine compound. The structural formula of the thienopyrimidine compound is shown as the following formula (I). The preparation process of the thienopyrimidine compound is simple and convenient, the steps of separation and purification are simple and feasible, and the yield of the product is high. The thienopyrimidine compound has antitumor activity on cervical cancer, colon cancer, breast cancer, lung cancer and liver cancer, and especially has higher activity on cervical cancer, colon cancer and breast cancer than colchicine, so that the thienopyrimidine compound has excellent antitumor effect and has application prospect.
Description
Technical Field
The present invention belongs to the field of heterocyclic compounds. More particularly relates to a thienopyrimidine compound, a preparation method and application thereof.
Background
Malignant tumor is a serious disease seriously jeopardizing human health, and is currently the disease with highest global mortality rate beyond cardiovascular disease, and the attack of malignant tumor is still a great challenge of modern medicine. In recent years, with the progress of research on cell carcinogenesis, various basic vital activity mechanisms in malignant tumor cells, such as signal transduction, cycle control, apoptosis induction, angiogenesis, etc., are gradually elucidated. However, although the existing clinical antitumor drugs have certain curative effects, the existing antitumor drugs still have the problems of poor selectivity, large toxic and side effects, easy occurrence of drug resistance and the like, so that the search for high-efficiency, high-selectivity and low-toxicity broad-spectrum antitumor drugs is a main direction of the development of the existing antitumor drugs.
The heterocyclic compounds containing nitrogen and sulfur occupy important positions in the design and screening of drug molecules, wherein thienopyrimidines are important nitrogen and sulfur heterocyclic compounds, are similar to purine in structure, have good anticancer, antimalarial, antibacterial, antioxidant, antiviral and other biological activities, and play pharmacological potential in the aspects of sterilization, disinsection, weeding and medicine; in addition, it is also widely used for research and screening of drug molecules. The compounds become hot spots for research of scientific researchers in the pharmaceutical field. The general synthetic methods of thienopyrimidines and their derivatives are: firstly, synthesizing thienopyrimidine derivatives from thiophene ring, and cyclizing with ortho-substituted aminothiophene and various electrophiles. Secondly, a thiophene ring is condensed from a pyrimidine ring to obtain a thienopyrimidine derivative, the method is relatively less applied, the halogen atom on the pyrimidine ring is replaced by a sulfhydryl group to introduce sulfur or the catalyst commonly used in the method is mainly transition metal Pd and hydroxide Pd (OH) 2 thereof through alkylation of thione on the pyrimidine ring.
Chinese patent CN200780024503.0 discloses a pharmaceutical compound based on a thienopyrimidine ring, which has activity as a PBK inhibitor and thus is useful for the treatment of diseases and disorders caused by abnormal cell growth, function or behaviour associated with PI3 kinase, such as cancer, immune disorders, cardiovascular diseases, viral infections, inflammation, metabolism/endocrine disorders and neurological disorders.
However, no approved antitumor drugs based on thienopyrimidine derivatives are reported at present. In addition, the anticancer effect of the existing thienopyrimidine compounds is still not ideal, and how to provide an acridine compound with excellent antitumor effect is a technical problem to be solved.
Disclosure of Invention
In view of the above-mentioned problems of the prior art, a primary object of the present invention is to provide a thienopyrimidine compound having excellent antitumor activity against cervical cancer, colon cancer, breast cancer, lung cancer and liver cancer, especially higher than colchicine in the antitumor activity against cervical cancer, colon cancer and breast cancer, and having excellent antitumor effect.
The second object of the invention is to provide a preparation method of the thienopyrimidine compound.
The third object of the invention is to provide the application of the thienopyrimidine compound in preparing antitumor drugs.
The fourth object of the invention is to provide an application of the thienopyrimidine compound and the PD-L1 inhibitor in combination in preparing anti-tumor drugs.
The above object of the present invention is achieved by the following technical solutions:
Thienopyrimidine compounds have a structural formula shown in the following formula (I):
Furthermore, the application also claims a preparation method of the thienopyrimidine compound shown in the formula (I), which comprises the following preparation steps:
(1) Dissolving 4-chloro-2-methylthiophene [3,2-d ] pyrimidine and 1-methyl-5-aminoindole in a first solvent, adding glacial acetic acid, reacting at 70-90 ℃, and carrying out suction filtration to obtain a 2-methyl-N- (1-methyl-1H-indol-5-yl) thieno [3,2d ] pyrimidine-4-amine crude product;
(2) Dissolving methyl iodide in a second solvent, mixing the second solvent with the 2-methyl-N- (1-methyl-1H-indol-5-yl) thieno [3,2d ] pyrimidine-4-amine crude product in the step (1), reacting at normal temperature under the catalysis of sodium hydride, and carrying out extraction post-treatment to obtain the thienopyrimidine compound;
The reaction formula of the preparation method is as follows:
preferably, the molar ratio of the 4-chloro-2-methylthiophene [3,2-d ] pyrimidine and the 1-methyl-5-aminoindole is 1:1-1.1.
Preferably, the molar ratio of the crude 2-methyl-N- (1-methyl-1H-indol-5-yl) thieno [3,2d ] pyrimidine-4-amine to the methyl iodide is 1:1-1.5.
Further preferred, the molar ratio of the crude 2-methyl-N- (1-methyl-1H-indol-5-yl) thieno [3,2d ] pyrimidin-4-amine to the methyl iodide is 1:1.5.
Preferably, the first solvent is absolute ethanol.
Preferably, the second solvent is anhydrous tetrahydrofuran.
Preferably, in the step (1), the reaction time is 1 to 3 hours.
Preferably, in the step (1), the reaction time is 2h.
Preferably, in the step (2), the reaction is performed at 20 to 25 ℃.
Preferably, in the step (2), the reaction time of the normal temperature reaction is 1 to 3 hours.
Further preferably, in the step (2), the reaction time of the normal temperature reaction is 2 hours.
Further preferably, in the step (1), glacial acetic acid is added and then reacted at 80 ℃.
Specifically, the post-extraction treatment is to extract the product obtained after the reaction in the step (2) by adopting a mixed solution of ethyl acetate and water, separate and obtain an organic phase, dry, desolventize, and separate and purify by adopting column chromatography to obtain the thienopyrimidine compound.
Specifically, as a specific embodiment of the present invention, the preparation method of the thienopyrimidine compound of formula (i) above comprises the following preparation steps: (1) Dissolving 4-chloro-2-methylthiophene [3,2-d ] pyrimidine and 1-methyl-5-aminoindole in absolute ethyl alcohol, adding glacial acetic acid, reacting for 1-3H at 70-90 ℃, and carrying out suction filtration to obtain a 2-methyl-N- (1-methyl-1H-indol-5-yl) thieno [3,2d ] pyrimidine-4-amine crude product;
(2) Dissolving methyl iodide in anhydrous tetrahydrofuran, mixing the anhydrous tetrahydrofuran with the 2-methyl-N- (1-methyl-1H-indol-5-yl) thieno [3,2d ] pyrimidine-4-amine crude product in the step (1), reacting for 1-3H at normal temperature under the catalysis of sodium hydride, extracting by adopting a mixed solution of ethyl acetate and water, separating to obtain an organic phase, drying, desolventizing, and separating and purifying by adopting column chromatography to obtain the thienopyrimidine compound.
In addition, the application also claims the application of the thienopyrimidine compound shown in the formula (I) in preparing antitumor drugs.
Furthermore, the application of the thienopyrimidine compound shown in the formula (I) and the PD-L1 inhibitor in combination in preparing anti-tumor drugs is also within the protection scope of the application. The inventors found through researches that, compared with the single use of the thienopyrimidine compound of formula (I) or the PD-L1 inhibitor, the combined use of the thienopyrimidine compound of formula (I) and the PD-L1 inhibitor has obvious synergistic effect, can exert more excellent anti-tumor effect in vivo and obtain more excellent anti-tumor effect.
Further preferably, the PD-L1 inhibitor is PD-L1 inhibitor NP-19.
Preferably, the tumor is one or more of cervical cancer, colon cancer, breast cancer, lung cancer or liver cancer.
More preferably, the tumor is one or more of cervical cancer, colon cancer or breast cancer.
Compared with the prior art, the invention has the following beneficial effects: the invention provides a novel thienopyrimidine compound, which has the advantages of simple preparation process, simple and easy separation and purification steps and high product yield. The thienopyrimidine compound has excellent anti-tumor activity for cervical cancer, colon cancer, breast cancer, lung cancer and liver cancer, and especially has higher anti-tumor activity for cervical cancer, colon cancer and breast cancer than colchicine, so that the thienopyrimidine compound has excellent anti-tumor effect and has application prospect.
Drawings
FIG. 1 is a nuclear magnetic resonance hydrogen spectrum of thienopyrimidines in example 1 of the present invention.
FIG. 2 is a nuclear magnetic resonance carbon spectrum of thienopyrimidines in example 1 of the present invention.
FIG. 3 is a graph showing tumor sizes in mice treated with different treatment groups.
FIG. 4 is a graph showing tumor growth inhibition of mice after treatment of mice with different treatment groups.
Figure 5 is a graph showing the change in weight of mice over time after treatment of mice with different treatment groups.
Detailed Description
The invention is further illustrated in detail below in connection with specific examples which are provided solely for the purpose of illustration and are not intended to limit the scope of the invention. The test methods used in the following examples are conventional methods unless otherwise specified; the materials, reagents and the like used, unless otherwise specified, are those commercially available.
EXAMPLE 1 preparation of thienopyrimidines
A thienopyrimidine compound comprising the following preparation steps:
(1) 4-chloro-2-methylthiophene [3,2-d ] pyrimidine (1 mmol,184.64 mg) and 1-methyl-5-aminoindole (1 mmol,146.19 mg) were dissolved in absolute ethanol, 1 drop glacial acetic acid was added, the reaction was completed for 30 minutes at 80 ℃, the solid was precipitated after the reaction, and the crude 2-methyl-N- (1-methyl-1H-indol-5-yl) thieno [3,2d ] pyrimidine-4-amine was obtained by suction filtration, and the next step was directly added.
(2) Methyl iodide is dissolved in anhydrous tetrahydrofuran, and is mixed with the crude product of 2-methyl-N- (1-methyl-1H-indol-5-yl) thieno [3,2d ] pyrimidine-4-amine obtained in the step (1), wherein the molar ratio of the methyl iodide to the crude product of 2-methyl-N- (1-methyl-1H-indol-5-yl) thieno [3,2d ] pyrimidine-4-amine is 1.5:1, under the catalysis of sodium hydride, reacting for 2 hours at normal temperature, extracting the reaction solution by using a mixed solution of ethyl acetate and water, drying an organic phase by using anhydrous sodium sulfate, desolventizing, separating and purifying by column chromatography to obtain 185mg of yellow solid. The overall yield of the process described in this example was calculated to be 60%.
Identifying the obtained yellow solid by nuclear magnetic resonance spectrum, as shown in figure 1 and figure 2, nuclear magnetic resonance hydrogen spectrum and nuclear magnetic resonance carbon spectrum of yellow solid product respectively, wherein the identification result is ：1H NMR(400MHz,CDCl3)δ7.61(s,1H),7.40(d,J＝8.5Hz,1H),7.33(d,J＝5.5Hz,1H),7.19(dd,J＝7.6,4.5Hz,3H),6.55(d,J＝2.5Hz,1H),3.89(s,3H),3.67(s,3H),2.71(s,3H).13C NMR(101MHz,CDCl3)δ163.42,161.11,158.04,136.22,135.46,133.14,130.45,128.62,123.23,122.84,122.05,113.29,109.96,101.63,77.41,77.09,76.77,39.74,33.06,25.84.
As shown by the identification result, the obtained yellow solid is N, 2-dimethyl-N- (1-methyl-1H-indol-5-yl) thieno [3,2d ] pyrimidine-4-amine, and the structural formula of the yellow solid is shown as the formula (I).
The preparation method of the synthetic thienopyrimidine compound has the following process route:
example 2 in vitro anti-tumor Activity assay of thienopyrimidines
In this example, the in vitro antitumor activity of the compound of formula (I) in example 1 was examined by MTT assay (the tumor cells used in this example were human cervical cancer cell HeLa, human colon cancer cell HCT116, human breast cancer cell MCF7, human lung cancer cell H23 and human liver cancer cell HepG 2).
Log phase cells were collected, cell suspension concentration was adjusted, inoculated into 96 well plates at 4×10 3-5×103/mL, and incubated for 12-24h. After cell attachment, different concentrations of the compound of formula (I) in example 1 were added and a total of 9 concentration gradients of 0.39,0.78,1.56,3.12,6.25,12.50,25,50,100nmol/L were set, 3 multiplex wells per concentration. The cells were placed in a 5% CO 2 incubator at 37℃to start time-lapse culture. After 48h of dosing, the 96-well plates were removed, and 20. Mu.L of 5mg/mL MTT solution was added to each well and incubation was continued for 4h at 37 ℃. Then, the supernatant in the wells was carefully aspirated, 100. Mu.L of DMSO was added to each well, and shaking was performed for 10min to dissolve the crystals. The absorbance (OD) of each well was measured on an ELISA detector at 570nm wavelength. Finally, data statistics is carried out, OD value (570 nm) is taken as a vertical axis, treatment time is taken as a horizontal axis, and inhibition effect of the drug on cell growth is depicted. The inhibition ratio was calculated as follows, inhibition ratio= (1-dosing group OD value/control group OD value) ×100%. Colchicine was set as positive control. The half-maximal inhibitory amount IC 50 was calculated using GRAPHPAD PRISM software. The results of the activities are shown in Table 1:
antitumor activity (nM), )
As can be seen from table 1 above, the antitumor activity of the compound of formula (i) in example 1 was significantly different from that of the positive control group for human cervical cancer cell HeLa with P < 0.05. For human colon cancer cells HCT116, the antitumor activity of the compound of formula (i) in example 1 was very significantly different from P <0.001 compared to the positive control group. For human breast cancer cells MCF7, the antitumor activity of the compound of formula (i) in example 1 was very significantly different from P <0.001 compared to the positive control group. In contrast, the antitumor activity of the compound of formula (I) in example 1 was slightly worse than that of the positive control group for human lung cancer H23 and human hepatoma cell HepG 2.
The in vitro experimental results show that the compound shown in the formula (I) in the embodiment 1 has stronger inhibition effect on five human tumor cells, namely human cervical cancer cells HeLa, human colon cancer cells HCT116, human breast cancer cells MCF7, human lung cancer cells H23 and human liver cancer cells HepG2, wherein the inhibition activity on human cervical cancer cells HeLa, human colon cancer cells HCT116 and human breast cancer cells MCF7 is obviously better than that of the positive control colchicine.
EXAMPLE 3 in vivo anti-tumor Activity assay of thienopyrimidines
The antitumor activity of the compounds of formula (I) in example 1 was tested using the following procedure, the specific test method being as follows:
B16-F10 tumor model of male C57 mice was established: the B16-F10 tumor cells were implanted subcutaneously in mice for about one week, and the mice were intraperitoneally injected with the compound of formula (I) in example 1 at a dose of 7mg/kg and PD-L1 inhibitor NP-19 at a dose of 7mg/kg once daily for 12 days. To determine whether the compound of formula (I) and PD-L1 inhibitor NP-19 in example 1 produced an antitumor synergy, mice were injected intraperitoneally with 3.5mg/kg of the compound of formula (I) and 3.5mg/kg of PD-L1 inhibitor NP-19 once daily for 12 days. A blank control group was set up in the experiment.
FIG. 3 is a graph showing tumor sizes in mice treated with different treatment groups; as shown in FIG. 3, the tumor size in mice was significantly smaller in the treatment group with 7mg/kg of the compound of formula (I) and the treatment group with 7mg/kg of the PD-L1 inhibitor, relative to the blank group. In the treatment group in which the compound of formula (I) was used in combination with the PD-L1 inhibitor, the tumor size in mice was significantly smaller than in the treatment group in which the compound of formula (I) was used alone, the treatment group in which the PD-L1 inhibitor was used, and the blank group.
FIG. 4 shows tumor growth inhibition of mice after treatment of mice with different treatment groups; as shown in FIG. 4, the compound of formula (I) of example 1 was effective in inhibiting the growth of B16-F10 tumor cells in mice with a Tumor Growth Inhibition (TGI) of 59.4%; mice in the PD-L1 inhibitor treated group had a Tumor Growth Inhibition (TGI) of 59.4%; the mice in the treatment group used the compound of formula (I) in combination with the PD-L1 inhibitor had a TGI of 73.1%. The treatment group with the compound of formula (i) and the PD-L1 inhibitor in combination showed a very significant difference of P <0.001 compared to the treatment group with the PD-L1 inhibitor alone, indicating that the compound of formula (i) and the PD-L1 inhibitor in combination could lead to an enhancement of antitumor activity. The treatment group with the compound of formula (i) and the PD-L1 inhibitor in combination showed significant differences with P <0.01 compared to the treatment group with the compound of formula (i) alone. The experimental group was compared with the blank control group, and the # # P <0.001 had a very significant difference.
Figure 5 is a graph showing the change in weight of mice over time after treatment of mice with different treatment groups. As shown in fig. 5, after 12 days of culture, the weight of the mice in the blank group was about 26g; in the treatment group with the compound of formula (I), the mice had a body weight of about 23g; in the treatment group with the PD-L1 inhibitor, the mice had a body weight of about 25g; in the treatment group with the compound of formula (I) in combination with the PD-L1 inhibitor, the mice had a weight of about 22.5g. The above data demonstrate that none of the three treatment groups of mice developed significant weight loss during the treatment period.
The result shows that the thienopyrimidine compound provided by the invention has excellent anti-tumor activity for cervical cancer, colon cancer, breast cancer, lung cancer and liver cancer, and especially has higher anti-tumor activity for cervical cancer, colon cancer and breast cancer than colchicine, so that the thienopyrimidine compound has excellent anti-tumor effect and good application prospect. Compared with the PD-L1 inhibitor or the thienopyrimidine compound which is used alone, the thienopyrimidine compound and the PD-L1 inhibitor have obvious synergistic effect and enhanced anti-tumor activity when being used in combination.
The above examples are preferred embodiments of the present invention, but the embodiments of the present invention are not limited to the above examples, and any other changes, modifications, substitutions, combinations, and simplifications that do not depart from the spirit and principle of the present invention should be made in the equivalent manner, and the embodiments are included in the protection scope of the present invention.
Claims (10)
1. The thienopyrimidine compound is characterized by having a structural formula shown in the following formula (I):
2. The method for preparing the thienopyrimidine compound of claim 1, which comprises the following preparation steps:
(1) Dissolving 4-chloro-2-methylthiophene [3,2-d ] pyrimidine and 1-methyl-5-aminoindole in a first solvent, adding glacial acetic acid, reacting at 70-90 ℃, and carrying out suction filtration to obtain a 2-methyl-N- (1-methyl-1H-indol-5-yl) thieno [3,2d ] pyrimidine-4-amine crude product;
(2) Dissolving methyl iodide in a second solvent, mixing the second solvent with the 2-methyl-N- (1-methyl-1H-indol-5-yl) thieno [3,2d ] pyrimidine-4-amine crude product in the step (1), reacting at normal temperature under the catalysis of sodium hydride, and carrying out extraction post-treatment to obtain the thienopyrimidine compound;
The reaction formula of the preparation method is as follows:
3. The process according to claim 2, wherein the molar ratio of 4-chloro-2-methylthiophene [3,2-d ] pyrimidine to 1-methyl-5-aminoindole is 1:1 to 1.1.
4. A process according to claim 2 or 3, wherein the molar ratio of crude 2-methyl-N- (1-methyl-1H-indol-5-yl) thieno [3,2d ] pyrimidin-4-amine to methyl iodide is from 1:1 to 1.5.
5. The method of claim 2, wherein the first solvent is absolute ethanol.
6. The method of claim 2, wherein the second solvent is anhydrous tetrahydrofuran.
7. The use of the thienopyrimidines in the preparation of anti-tumor drugs according to claim 1, characterized in that the tumor is one or more of cervical cancer, colon cancer, breast cancer, lung cancer or liver cancer.
8. The use of a thienopyrimidine compound and a PD-L1 inhibitor in combination for the preparation of an antitumor drug according to claim 1, wherein the tumor is one or more of cervical cancer, colon cancer, breast cancer, and melanoma.
9. The use according to claim 8, wherein the melanoma is melanoma cells B16-F10.
10. The use according to claim 8, wherein the PD-L1 inhibitor is PD-L1 inhibitor NP-19.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202211520649.7A CN115746017B (en) | 2022-11-30 | Thienopyrimidine compound and preparation method and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202211520649.7A CN115746017B (en) | 2022-11-30 | Thienopyrimidine compound and preparation method and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN115746017A CN115746017A (en) | 2023-03-07 |
| CN115746017B true CN115746017B (en) | 2024-06-07 |
Family
ID=
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2003074529A2 (en) * | 2002-03-01 | 2003-09-12 | Pfizer Inc. | iNDOLYL-UREA DERIVATIVES OF THIENOPYRIDINES USEFUL AS ANTI-ANGIOGENIC AGENTS |
| CN101328186A (en) * | 1997-11-11 | 2008-12-24 | 辉瑞产品公司 | Thienopyrimidine and thienopyridine derivatives useful as anticancer agents |
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101328186A (en) * | 1997-11-11 | 2008-12-24 | 辉瑞产品公司 | Thienopyrimidine and thienopyridine derivatives useful as anticancer agents |
| WO2003074529A2 (en) * | 2002-03-01 | 2003-09-12 | Pfizer Inc. | iNDOLYL-UREA DERIVATIVES OF THIENOPYRIDINES USEFUL AS ANTI-ANGIOGENIC AGENTS |
Non-Patent Citations (1)
| Title |
|---|
| Design and SAR of thienopyrimidine and thienopyridine inhibitors of VEGFR-2 kinase activity;Michael J. Munchhof，等;《Bioorganic & Medicinal Chemistry Letters》;第14卷;第21–24页 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CA2930203A1 (en) | N-benzyl tryptanthrin derivative, and preparation method and application thereof | |
| CN111393404B (en) | Benzothiophene compounds, and pharmaceutical composition and application thereof | |
| CN107417695B (en) | Berberine derivatives, preparation method thereof, pharmaceutical composition and anti-tumor application thereof | |
| CN101012227A (en) | Novel 13-n-octylberberine derivative with antineoplastic action | |
| CN108101910B (en) | N-substituted pyrazolo [3,4-d ] pyrimidone compound and preparation method and application thereof | |
| CN115746017B (en) | Thienopyrimidine compound and preparation method and application thereof | |
| CN101591226A (en) | 1,3-diaryl propane analog derivative and uses thereof | |
| CN106187923A (en) | 2 aryl 4 aroyl triazole compounds and application thereof | |
| CN115746017A (en) | Thienopyrimidine compound and preparation method and application thereof | |
| CN101817829A (en) | 4-substituted anilino-podophyllotoxine derivative and application | |
| CN101798306B (en) | Application of uncaria alkaloid monomer | |
| KR102606167B1 (en) | Fluorine-containing substituted benzothiophene compounds, pharmaceutical compositions and applications thereof | |
| CN110283192B (en) | Preparation method and application of tryptanthrin derivative containing boric acid | |
| CN111116551B (en) | 1-azaspiro [5.5] undecane-3-ones and 1-azaspiro [5.5] undecane-3-ols | |
| CN110172058B (en) | 7-azaspiro [5.6] dodecane-10-one compound and preparation method and application thereof | |
| CN114478561A (en) | Epalrestat lycorine conjugate and preparation method and application thereof | |
| CN108484617B (en) | Novel benzofuran azanaphthalene dione derivative and preparation method thereof | |
| AU640831B2 (en) | Process for preparing benzo(c)phenanthridinium derivatives, and novel compounds prepared by said process | |
| CA2271940C (en) | Novel phenanthridinium derivatives | |
| CN111892581A (en) | Quinazoline derivative with anti-tumor activity and synthesis method and application thereof | |
| CN115073432B (en) | Methylated flavanol alkaloid and extraction, synthesis, detection and application thereof | |
| CN108904503B (en) | Application of 6-chloro-5-nitro-2, 4-diaminopyrimidine in medicine for treating chronic granulocytic leukemia | |
| CN116239594B (en) | 6- (imidazo [1,2-a ] pyridin-6-yl) quinazoline derivatives and uses thereof | |
| CN114853736B (en) | Compound with TRK inhibitory activity, preparation method, composition and application thereof | |
| CN117003766B (en) | Anti-influenza virus derivative and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant |