CN103242341B - Thieno-2,4 substituted pyrimidines compounds and pharmaceutical composition thereof and application - Google Patents

Abstract

The invention discloses a thieno 2,4 substituted pyrimidine compound with the general formula (I), (II), (III) and (IV) or a pharmaceutically acceptable salt or stereoisomer or prodrug thereof Molecules and their pharmaceutical compositions and applications. The above-mentioned thieno-2,4-substituted pyrimidine compounds can effectively inhibit the abnormal expression of Aurora kinase, have special inhibitory effects on Aurora-A and Aurora-B, and can be applied to the new field of molecular targeted therapy, especially for hyperproliferative diseases. It has strong inhibitory activity on cervical tumor cells, human macrophages, leukemia cells, and human T lymphocytes.

Description

Thieno 2,4 substituted pyrimidine compounds and their pharmaceutical composition and application

technical field

The invention relates to the field of chemistry and medicine, in particular to thieno 2,4 substituted pyrimidine compounds and their pharmaceutical composition and application.

Background technique

Tumor molecular targeted therapy is a treatment method based on the selective killing of tumor cells by chemical or biological means to key molecules closely related to tumor growth. The characteristics of targeted therapy are: high specificity, strong selectivity, and mild side effects; when used in combination, it can strengthen the curative effect of traditional chemotherapy and radiotherapy, and reduce postoperative recurrence. Gleevec (STI571, imatinib, commodity The targeted drug represented by Gleevec has ushered in a new era for cancer chemotherapy. Tumor targeted therapy has developed rapidly in just a few years. The emergence of tumor targeted therapy has had an impact on traditional drug administration concepts and modes. For example, due to the low toxicity and side effects, targeted drugs often fail to achieve dose-limiting toxicity and maximum tolerated dose in Phase I clinical trials; Satisfactory curative effect can be achieved without the maximum tolerated dose. Therefore, tumor targeted therapy is a hot spot and development trend of tumor therapy.

Aurora kinases are a group of three highly homologous serine-threonine protein kinases that play important regulatory roles in mitosis. The Aurora family is divided into AuroraA, AuroraB, and AuroraC. From their discovery in 1995 to their first application in 1998 to observe the expression of human cancerous tissues, these kinases have become the subject of intense research in both academic and production aspects of the oncology industry. This effort has achieved fruitful results. So far, more than 10 Aurora kinase inhibitors have passed the early clinical evaluation. These characteristically characterized complexes are highly selective relative to most other kinases, and most of them are cross-reactive, a small set of kinases relevant to tumor biology, most notably Representatives of Abl and Flt-3 kinases.

Aurora kinase inhibitors can be subdivided into three general classes: those possessing selectivity for Aurora-A over Aurora-B, those possessing selectivity for Aurora-B over Aurora-A, the third The second protein inhibitor possesses selectivity to both Aurora-A and Aurora-B.

Aurora kinases and mitosis

Aurora-A: Aurora-A is involved in the regulation of early mitotic activity and is also involved in mitosis. Protein expression and kinase activity rise during the G2 phase of the cell cycle and peak in early mitosis (Meraldi et al., 2004; Carmena and Earnshaw, 2003; Andrew et al., 2003). From frequent observations of monopolar spindles, it was found that depletion of Aurora-A resulted in a delay in participating in mitosis and marking the division of the spindle. At the molecular level, the role of Aurora-A in the rest of mitosis is fully elucidated; however, there are some conclusive findings that may help explain the inhibition of Aurora-A in some data and help identify its specific biomarkers.

Aurora-B: Aurora-B is a component that catalyzes the synthesis of chromosome passenger complex (CPC), which determines the continuous progress and completion of mitosis. This complex consists of four proteins, namely Aurora-B, survivin, INCENP and borealin. The non-catalytic components of the complex act in part to regulate confinement and activate Aurora-B. Depletion of any component of this complex can negatively affect the other components and disrupt mitotic progression. Survivin, INCENP, and borealin are all known substrates for Aurora-B. Although the effects of survivin and borealin on phosphorylation are unknown, in the case of INCENP, it seems to play a role in the feedback loop. Positive effects (Andrew et al., 2003; Carmena and Earnshaw, 2003; Meraldi et al., 2004).

Aurora-C: Aurora-C is the least studied of the Aurora kinases, and its specific role in mitosis is not well defined. Aurora-C is significantly inferior to Aurora-A or Aurora-B in most somatic tissues at lower levels. High expression of Aurora-C in testis tissue was the only exception. Consistent with this expression profile, experiments in homozygous versus heterozygous mice identified its critical role in nuclear meiosis. These mice exhibit a very normal physiology, but also have low fertility caused by defective sperm (ZhouH, KuangJ, ZhongL, et.al, Nat.Gent, 1998, 20(2):189-193; LittlepayLE, WuH , AndressonT, et.al, ProcNatlAcadSciUSA, 2002, 99(24):15440-15445), these are related to aberrant chromosome condensation and polyploidy.

Aurora kinases and cancer cells

Since its discovery in the 1990s, a wealth of scientific and clinical data has shown a strong link between Aurora kinases and human cancer progression.

Aurora-A has 20q13.2 (ZhouH, KuangJ, ZhongL, et.al, Nat.Gent, 1998, 20(2):189-193) genome domain, which is a gene map closely related to human cancer. Overexpression of Aurora-A protein was observed in many cancer cells. Ectopic overexpression of Aurora-A in vitro results in cells displaying multiple cancer cell features such as centrosome amplification, aneuploidy, chromosomal instability, and telomere elongation. It has been observed that Aurora-A expresses itself, or activates its partner TPX-2 (Hirota et al., 2003; Bayliss et al., 2003; EyesandMaller, 2004; Kufer et al., 2002; Satinover et al., 2004), which associates with human cancer cell chromosomes related to instability. Furthermore, Aurora-A is associated with and negatively regulates a major determinant of tumor suppression. Perhaps the most striking of these is p53, which degenerates and represses its transcriptional activity as Aurora-A promotes mdm2-mediated degradation. Regardless, Aurora-A does not insist that cells transformed and overexpressed in vitro are not in the usual active tumor cells and are therefore not a regular protein. Taken together, these observations suggest that Aurora-A generally requires otherwise unregulated oncogenes to be transformed. The potential implication of this is that selective inhibition of Aurora-A may only show anticancer activity against specific cancer cells or be best used in combination with other drugs. This appears to be in stark contrast to the aforementioned attrition studies in which removal of Aurora-A frequently resulted in major defects in mitosis. The reason for this discrepancy may be due to the relatively small number of cancer cell lines used in these studies. It is likely that the true impact of Aurora-A inhibitors has only been revealed once and selective inhibition has been widely used in cancer cells.

Consistent with the hypothesis, the possible benefit of Aurora-A inhibition in combination with other therapies has been demonstrated by certain groups, and Aurora-A is depleted in sensitive cancer cells due to chemotherapeutic agents such as taxanes, cisplatin, and ionizing radiation. The role of cytotoxins produced.

The gene map of Aurora-B in the 17p13.1 genomic region has changes in some human cancer regions (ZengWF, NavaratneK, PraysonRA, et.al, JClinPathol, 2007, 60(2):218-221). Aurora-B mRNA and protein itself are frequently overexpressed in cancer and in tumors with some reported types of serious disease in terms of protein expression. In addition to these direct links between Aurora-B and cancer, it is important to note that the CPC protein, which is expressed in cooperation with or regulated by Aurora-B, is also often overexpressed or exaggerated for cancer. Although the key to cancer is not very tightly linked to Aurora-B during cell division, it is not transcribed in vitro and does not generally form tumors in vivo. An important exception is the tumorigenic behavior where Aurora-B is already overexpressed on mutant forms of p53 cells (ManfrediMG, EcsedyJA, MeetzeKA, ProcNatlAcadSciUSA, 2007, 104(10):4106-4111.). The results from these experiments suggest that although Aurora-B is not an oncogene itself, it can act as a partner in tumor formation with mutations in other oncogenes. Consistent with this notion is that Aurora-B enhances Ras-mediated conversion, but depletion of Aurora-B with short RNAs inhibits Ras conversion. Further research is needed on Aurora-B as an important anticancer target, including Aurora-B depletion in sensitive cancer cell therapies, such as the cytotoxic effects of alkylating agents and ionizing radiation.

Recent data suggest that the roles of Aurora-B and Aurora-C overlap in many mitotic processes. Although Aurora-C has been observed in a number of human cancer cell lines, a clear role for Aurora-C in tumorigenesis has not yet been established.

Aurora Kinase Small Molecule Inhibitors as Chemical Probes

Depletion of Aurora kinase activity using techniques such as siRNA and expression of kinase-inactivating proteins helps to predict the biological profile of Aurora inhibitors (MorrowCJ, TigheA, JohnsonVL, et.al, JCellSci, 2005, 118(16): 3639-3652 ) inhibitors are required. The following are the chemical structural formulas of the published Aurora inhibitors (complexes 1-6).

Dual inhibitor of Aurora-A and Aurora-B. In 2003, the first small molecule inhibitors of Aurora were disclosed (ZM4474391 with hesperidin 2). Complex 1 was able to inhibit Aurora-A and Aurora-B more efficiently than other unrelated kinases (excluding the closely related Aurora-C) ( _IC50 at 110nm and 130nm, respectively) (MorlockAA, KeenNJ, JungFH, et al. .al, WO0121596[P], 2001-03-29). This selectivity strongly supports its wide application as a chemical probe. Complex 2 is said to be an inhibitor of Aurora-B (IC ₅₀ at 250nm), possessing significant cross-reactivity against six other kinases (no data indicating that this includes Aurora-A or Aurora-C). Both inhibitors show similar full dominance on polyploid cells: inhibition of histone H3 phosphorylation (Ser10) and endoreduplication in the absence of cell division (Ota et al., 2002). Observed in some cases of giant cells with as many as 32 DNA replications. An experiment showed that a cell G1 was blocked in complex 1 and failed to complete cytoplasmic division. Under these conditions, entry and exit from mitosis proceed normally, but cells fail to divide. Inhibitors of these two enzymes cause marked chromosomal dysregulation, often juxtaposed with chromosomes on the spindle of nonhermaphroditic microtubules. Despite these aberrations, the separation of the two sister chromatids persists. Taken together, these activities strongly suggest that inhibition of Aurora kinase activity leads to the abrogation of the spindle checkpoint and exit of mitosis for chromosome attachments under appropriate circumstances. This was confirmed by the appearance of captive mitotic arrest of these two complexes induced by taxane-type microtubule stabilizers. More detailed analysis revealed that components of many spindle checkpoints, including BurR1, Mad2 and CENP-E, were not fixed. Not surprisingly, complex 1-treated cells had reduced viability as observed by defective mitosis.

The characterization of complex 3 as a potential Aurora kinase inhibitor in vivo was first disclosed in 2004. Complex 3 (MK-0457(VX-680)) is a potent inhibitor potent against all three Aurora kinases while being selective for others (HarringtonEA, BebbingtonD, MooreJ, et.al, Nat. Med, 2004, 10(3):262-267). Consistent with dual inhibitors of Aurora-A and Aurora-B (complexes 1 and 2), complex 3 rapidly induces polyploidy and arrests proliferation. Most notably, random mitosis mediated by Aurora is sufficient to cause apoptosis. This case is the entire cell cycle processed by complex 3 including cancer cell lines, non-cancer cell lines and primary tumor cell sampling. Corresponding to this mechanism of cell death in the cell cycle is the observation that cells in non-cycling phases do not lose viability. Complex 3 is very effective in blocking tumor growth and even causing some tumor regression in rodent models of human cancer (Wilkinson RW, Odedra R, Hcaton SP, et. al, Clin Cancer Res, 2007, 13(12):3682-3688). Immunohistochemical analysis of complex 3 revealed significant inhibition of tumor treatment histone H3 phosphorylation, consistent with Aurora-B inhibition, leading to a marked reduction in apoptosis. Effective doses of complex 3 were well tolerated, but a significant and reversible decrease in neutrophil numbers (>50% nadir) was observed. Clinical observations have found that Aurora kinase has an inhibitory effect on general neutrophils based on the mechanism and form of non-cancerous cell cycle. Complex 3 can induce the formation of monopolar spindles, a phenotype that follows Aurora-A depletion and has no reported replacement for complex 1, a dual inhibitor of Aurora-A and Aurora-B. Thus, complex 3 is able to inhibit specific Aurora-A substrate phosphorylation in addition to Aurora-B substrate histone H3. Thus, complex 3 can inhibit both Aurora-A and Aurora-B in terms of mitosis and cell viability.

Although it has been widely reported that Aurora kinase inhibitors cause mitotic abnormalities and arrest cycle cell proliferation, several research groups now demonstrate that the ultimate fate of cells seems to be related to the genetic background. In cells treated with complex 1 or 3, cells lacking p53 gene function undergo proliferation and massive endoreplication followed by cytokine deficiency followed by cell death. On the other hand, cells with full p53 function were less likely to endoreplicate. This result suggests a role for p53 at the G1 tetraploid checkpoint. This will make it interesting to see whether this condition can be observed clinically.

The emergence of this mechanism will lead to the frequent occurrence of tumor chemotherapy resistance. In many cases, resistance is associated with the expression of drug pumps, but can be observed in mutations that target the drug itself. A small number of polymorphisms in the human Aurora gene have been observed, although these effects on kinase activity or on drug sensitivity are not typical. A recent study showed that cancer cells cultured in the long-term presence of complex 1 can clonal against this complex. Detailed analysis revealed that this resistance was due to a number of mutations in the Aurora-B protein. Expression of these mutant Aurora-B proteins has cellular effects beyond complex 1 on histone H3 phosphorylation, cell cycle, and spindle checkpoints. This clearly indicates that the antimitotic effect of complex 1 is related to the inhibition of Aurora-B rather than Aurora-A. These Aurora-B mutations also block complex 3 cell cycle and colony formation. Again, the anticancer properties of complex 3 are related to the inhibition of Aurora-B rather than Aurora-A or Aurora-C. This is somewhat unexpected compared to the effect of Aurora-A depletion on antimitosis. This shows that in this experiment, complexes 1 and 3 cannot completely inhibit Aurora-A, or inhibit the catalytic activity of Aurora-A, and only have a weak effect on itself. The statistical results of these observations found that Aurora-A usually does not transform or tumorize, and the conclusion is that Aurora-A may produce the best curative effect when combined with other cancer therapies. Of course, more importantly, this study highlights the potential clinical resistance of Aurora kinase inhibitors, which will allow Aurora kinase inhibitors to be more strictly monitored through clinical practice.

Beyond the effects of individual drugs on dual inhibitors of Aurora kinases, several groups have shown that these complexes can make cells more sensitive to other treatments. The effects of dexamethasone, streptomycin, etoposide, vincristine, daunorubicin, and ionizing radiation were all enhanced when evaluated in combination with Aurora-A/B dual inhibitors (YangJ, IkezoeT, NishiokaC, et.al, Blood, 2007, 110(6):2034-2040). While some of these combined effects can be attributed to inhibition of key mitotic processes by Aurora kinases, the mechanism for the synergy observed in other cases is unclear. However, these clinical observations have the potential to have a large impact on the design of clinical studies.

The recent elucidation of the biological functions of Aurora kinases in normal and cancer cells has led to the rapid development of small molecule inhibitors. Preliminary results of preclinical and clinical trials show that although the molecular mechanism of Aurora kinase inhibitors inducing growth arrest and apoptosis of cancer cells remains to be further explained, such substances are very promising for the treatment of cancer. Therefore, in the process of treating cancer in the future. Aurora kinase inhibitors need to be further confirmed as a powerful anticancer strategy, and many clinical trials will give answers in the near future.

Pyrimidines are generally known as inhibitors of kinases. For example, substituted pyrimidines with non-aromatic groups at the 4-position are described as active ingredients with anticancer effects in International Patent Applications WO021096888 and WO031032997, and are used as Aurora inhibitors 2,4-Diamino-pyrimidines are described in International Patent Application WO2007/003596. In fact, most of the current Aurora kinase inhibitor anti-tumor drugs have drug-induced drug-resistant gene mutations and face problems such as narrow clinical application. Therefore, it is of great significance to develop more novel protein Aurora kinase inhibitors to overcome existing drug resistance and improve clinical efficacy.

Contents of the invention

Based on this, the present invention expands the scope of pyrimidine compounds, and synthesizes more novel active compounds for preventing and/or treating diseases characterized by excessive or abnormal cell proliferation. The object of the present invention is to provide a new active substance-thieno 2,4 substituted pyrimidines which can be used for the prevention and/or treatment of excessive or abnormal cell proliferation.

A thieno 2,4 substituted pyrimidine compound or its pharmaceutically acceptable salt or stereoisomer or prodrug molecule, the general formula is:

Wherein, X ₁ and X ₂ are simultaneously or independently O, S, S(O), S(O) ₂ or NR ₅ , wherein R ₅ is H or C _1-6 alkyl;

X ₃ and X ₄ are simultaneously or independently O, N, CH, m, n, o, p are integers between 1-4;

R ₁ and R ₂ are simultaneously or independently selected from the following groups:

replaced or not replaced

Wherein, the dotted line represents the bond connected with X ₁ and X ₂ ;

a is an integer between 0-3, and X, Y, and Z are simultaneously or independently N, CH;

R ₆₁ , R ₆₂ , R ₆₃ are simultaneously or independently hydrogen, C _1-5 alkyl, halo, nitro, cyano, amino or hydroxyl;

R ₃ and R ₄ are simultaneously or independently selected from the following groups:

Hydrogen, C _1-5 alkyl, C _3-8 cycloalkyl or R ₆ COR ₇ ; wherein, R ₆ is C _1-3 alkyl or C _1-3 alkylphenyl, R ₇ is C _1-5 Alkoxy, halo, amino, hydroxyl, C _1-5 alkyl substituted phenyl, C _1-5 alkyl substituted heterocyclic aryl.

In the above-mentioned thieno2,4-substituted pyrimidine compounds or pharmaceutically acceptable salts or stereoisomers or prodrug molecules, "alkyl" means to include branched and linear saturated groups with a specific number of carbon atoms. Aliphatic hydrocarbon group. For example, the definition of "C _1-5 " in "C _1-5 alkyl" includes groups having 1, 2, 3, 4 or 5 carbon atoms arranged in a linear or branched chain. For example, "C _1-5 alkyl" specifically includes methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, isobutyl, pentyl. The term "cycloalkyl" refers to a monocyclic saturated aliphatic hydrocarbon group having the specified number of carbon atoms. For example "cycloalkyl" includes cyclopropyl, methyl-cyclopropyl, 2,2-dimethyl-cyclobutyl, 2-ethyl-cyclopentyl, cyclohexyl and the like.

"Heterocycle" or "heterocyclyl" refers to a 5-6 membered aromatic or non-aromatic heterocycle containing 1-4 heteroatoms selected from O, N and S, and includes bicyclic groups. "Heterocyclyl" includes heteroaryl, as well as dihydrogenated and tetrahydrogenated analogs thereof. Further examples of "heterocyclyl" include, but are not limited to: imidazolyl, indolyl, isothiazolyl, isoxazolyl, oxadiazolyl, oxazolyl, oxetanyl, pyranyl Base, pyrazinyl, pyrazolyl, pyridazinyl, pyridyl, pyrimidinyl, pyrrolyl, quinoxalinyl, tetrazolyl, thiadiazolyl, thiazolyl, thienyl, triazolyl, 1,4 -Dioxanyl, pyrrolidinyl, dihydroimidazolyl, dihydroisoxazolyl, dihydroisothiazolyl, dihydrooxadiazolyl, dihydrooxazolyl, dihydropyrazinyl, dihydropyridine Azolyl, dihydropyridyl, dihydropyrimidinyl, dihydropyrrolyl, dihydrotetrazolyl, dihydrothiadiazolyl, dihydrothiazolyl, dihydrothienyl, dihydrotriazolyl, dihydronitrogen Heterobutanyl, tetrahydrofuranyl and tetrahydrothienyl, and N-oxides thereof. Attachment of heterocyclic substituents can be via carbon atoms or via heteroatoms.

"Halo" is meant to include chlorine, fluorine, bromine and iodine.

Unless otherwise defined, alkyl, cycloalkyl, aryl, heteroaryl, and heterocyclyl substituents can be unsubstituted or substituted. For example, the H atom on _C1-5 alkyl or cycloalkyl can be replaced by one, two or three selected from hydroxyl, halo, alkoxy, dialkylamino or heterocyclic groups such as morpholinyl, piper substituents such as pyridyl and the like.

In the above-mentioned thieno-2,4-substituted pyrimidine compounds or pharmaceutically acceptable salts or stereoisomers or prodrug molecules thereof, when any variable (such as R ₁ , R ₂ , etc.) appears more than once in any component, Then the definition of each occurrence of it is independent of the definition of each other occurrence. Also, combinations of substituents and variables are permissible only if such combinations render the compounds stable. Knowing the art, one of ordinary skill can select substituents and substitution patterns for the compounds of the present invention to provide compounds that are chemically stable and readily synthesized from readily available starting materials by skill in the art and by methods set forth below. If a substituent is itself substituted with more than one group, it is understood that these groups may be on the same carbon atom or on different carbon atoms, so long as the structure is stabilized.

The present invention includes free forms of compounds of formulas I, II, III, and IV, as well as pharmaceutically acceptable salts and stereoisomers thereof.

Pharmaceutically acceptable salts of the compounds of the present invention include conventional non-toxic salts of the compounds of the present invention formed by reacting the compounds of the present invention with inorganic or organic acids. For example, conventional nontoxic salts include those derived from inorganic acids such as hydrochloric, hydrobromic, sulfuric, sulfamic, phosphoric, nitric, and the like, as well as organic acids such as acetic, propionic, succinic, glycolic, hard Fatty acid, lactic acid, malic acid, tartaric acid, citric acid, ascorbic acid, pamoic acid, maleic acid, hydroxymaleic acid, phenylacetic acid, glutamic acid, benzoic acid, salicylic acid, sulfanilic acid, 2-acetyl Oxy-salts prepared from benzoic acid, fumaric acid, toluenesulfonic acid, methanesulfonic acid, ethanedisulfonic acid, oxalic acid, isethionic acid, trifluoroacetic acid, etc.

The preparation of the above-mentioned pharmaceutically acceptable salts and other typical pharmaceutically acceptable salts is described in detail by Berg et al. in Pharmaceutical Salts, J. Pharm. Sci. 1977, 66: 1-19.

In one of the embodiments, the At least one hydrogen atom on the ring is replaced by a substituent, and the substituent is C _1-5 alkyl, C _1-3 alkoxy, halogen, aryl, saturated or unsaturated heterocyclic group, The saturated heterocyclic group is morpholinyl, thiomorpholinyl, pyrrolidinyl, piperazinyl, piperidinyl, imidazolidinyl or pyrazolidinyl; the aryl, saturated or unsaturated heterocyclic group Both are connected with one or two substituents selected from the following groups: hydroxyl, halo, C _1-3 alkyl, C _1-3 alkoxy, C _1-3 alkanoyloxy, trifluoromethane Group, cyano group, amino group, nitro group, C _1-4 alkoxycarbonyl group.

In one of the embodiments, the C _1-5 alkyl or C _3-8 cycloalkyl is a C _1-5 alkyl or C _3-8 cycloalkyl with at least one H atom replaced by a hydroxyl, fluorine atom or amino group alkyl.

In one embodiment, the X ₁ and the X ₂ are simultaneously or independently NH or NCH ₃ .

In one of the embodiments, the R ₁ and R ₂ are simultaneously or independently selected from the following structures:

Wherein, both e and f are integers between 0-3.

In one of the embodiments, the R ₁ and R ₂ are simultaneously or independently

In one embodiment, both X ₁ and X ₂ are N, X ₃ and X ₄ are N, O or CH simultaneously or independently, and R ₁ and R ₂ are simultaneously or independently selected from the following structures:

Wherein, g is an integer between 0-3.

In one of the embodiments, one of the following compounds is selected from:

N ² -(4-fluoro-3-(trifluoromethyl)benzyl)-N ⁴ -(5-methyl-1H-pyrazole-3-substituted)thieno[3,2-d]pyrimidine-2 , 4-diamine;

N ⁴ -(5-methyl-1H-pyrazole-3-substituted)-N ² -(4-methylbenzyl)thieno[3,2-d]pyrimidine-2,4-diamine;

N ⁴ -(5-methyl-1H-pyrazole-3-substituted)-N ² -(3-methylbenzyl)thieno[3,2-d]pyrimidine-2,4-diamine;

N ² -(3-fluorobenzyl)-N ⁴ -(5-methyl-1H-pyrazole-3-substituted)thieno[3,2-d]pyrimidine-2,4-diamine;

N ² -(3-bromobenzyl)-N ⁴ -(5-methyl-1H-pyrazole-3-substituted)thieno[3,2-d]pyrimidine-2,4-diamine;

N ² -(4-bromo-2-fluorobenzyl)-N ⁴ -(5-methyl-1H-pyrazole-3-substituted)thieno[3,2-d]pyrimidine-2,4-diamine ;

2-(4-fluorophenyl)-1-(4-(4-(5-methyl-1H-pyrazole-3-substituted amine)thieno[3,2-d]pyrimidine-2-substituted)piper pyridine-1-substituted) ethyl ketone;

N ² -Benzhydryl-N ⁴ -(5-methyl-1H-pyrazole-3-substituted)thieno[3,2-d]pyrimidine-2,4-diamine;

N ² -(1-benzylpiperidine-4-substituted)-N ⁴ -(5-methyl-1H-pyrazole-3-substituted)thieno[3,2-d]pyrimidine-2,4-di amine;

N ² -((2-chloropyridine-4-substituted)methyl)-N ⁴ -(5-methyl-1H-pyrazole-3-substituted)thieno[3,2-d]pyrimidine-2,4 - diamines;

N ² -(Benzo[d][1,3]dioxolane-5-substituted methyl)-N ⁴ -(5-methyl-1H-pyrazole-3-substituted)thieno[3,2 -d] pyrimidine-2,4-diamine;

N ² -(2-fluorobenzyl)-N ⁴ -(5-methyl-1H-pyrazole-3-substituted)thieno[3,2-d]pyrimidine-2,4-diamine;

(3-chlorophenyl)(4-(4-(5-methyl-1H-pyrazole-3-substituted amine)thieno[3,2-d]pyrimidine-2-substituted)piperazine-1-substituted ) methyl ketone;

N ² -(4-chlorobenzyl)-N ⁴ -(5-methyl-1H-pyrazole-3-substituted)thieno[3,2-d]pyrimidine-2,4-diamine;

N ² -(2-chloro-6-fluorobenzyl)-N ⁴ -(5-methyl-1H-pyrazole-3-substituted)thieno[3,2-d]pyrimidine-2,4-diamine ;

N ² -(2,6-dichlorobenzyl)-N ⁴ -(5-methyl-1H-pyrazole-3-substituted)thieno[3,2-d]pyrimidine-2,4-diamine;

N ⁴ -(5-methyl-1H-pyrazole-3-substituted)-N ² -(4-(trifluoromethyl)benzyl)thieno[3,2-d]pyrimidine-2,4-di amine;

N ² -(4-chlorophenethyl)-N ⁴ -(5-methyl-1H-pyrazole-3-substituted)thieno[3,2-d]pyrimidine-2,4-diamine;

N ² -(2,4-difluorophenyl)-N ⁴ -(5-methyl-1H-pyrazole-3-substituted)thieno[3,2-d]pyrimidine-2,4-diamine;

(4-chlorophenyl)-(4-(4-(5-methyl-1H-pyrazole-3-substituted amine)thieno[3,2-d]pyrimidine-2-substituted)piperidine-1- substituted) methyl ketone;

(5-chloro-2-fluorophenyl)-(4-(4-(5-methyl-1H-pyrazole-3-substituted amine)thieno[3,2-d]pyrimidine-2-substituted)piper pyridine-1-substituted)methanone;

2-(2-chlorophenyl)-1-(4-(4-(5-methyl-1H-pyrazole-3-substituted amine)thieno[3,2-d]pyrimidine-2-substituted)piper pyridine-1-substituted) ethyl ketone;

(4-(4-(5-methyl-1H-pyrazole-3-substituted amine)thieno[3,2-d]pyrimidine-2-substituted)piperidine-1-substituted)(1-phenyl ring Propanyl) ketone;

N ² -(3-bromophenyl)-N ⁴ -(5-methyl-1H-pyrazole-3-substituted)thieno[3,2-d]pyrimidine-2,4-diamine;

(4-bromophenyl)-(4-(4-(5-methyl-1H-pyrazole-3-substituted amine)thieno[3,2-d]pyrimidine-2-substituted)piperidine-1- substituted) methyl ketone;

N ⁴ -(5-methyl-1H-pyrazole-3-substituted)-N ² -(3-morpholinopropyl)thieno[3,2-d]pyrimidine-2,4-diamine;

N ² -(biphenyl-4-substituted)-N ⁴ -(5-methyl-1H-pyrazole-3-substituted)thieno[3,2-d]pyrimidine-2,4-diamine;

N-(5-methyl-1H-pyrazole-3-substituted)-2-(3-methylpiperazine-1-substituted)thieno[3,2-d]pyrimidin-4-amine;

N ² -(4-chloro-3-(trifluoromethyl)phenyl)-N ⁴ -(5-methyl-1H-pyrazole-3-substituted)thieno[3,2-d]pyrimidine-2 , 4-diamine;

N ² -(4-bromo-3-fluorophenyl)-N ⁴ -(5-methyl-1H-pyrazole-3-substituted)thieno[3,2-d]pyrimidine-2,4-diamine ;

N ² -(3-fluoro-5-(trifluoromethyl)phenyl)-N ⁴ -(5-methyl-1H-pyrazole-3-substituted)thieno[3,2-d]pyrimidine-2 , 4-diamine;

N-(5-methyl-1H-pyrazole-3-substituted)-2-(piperazine-1-substituted)thieno[3,2-d]pyrimidin-4-amine;

N ² -(2,6-diethylphenyl)-N ⁴ -(5-methyl-1H-pyrazole-3-substituted)thieno[3,2-d]pyrimidine-2,4-diamine ;

(4-(4-(5-methyl-1H-pyrazole-3-substituted amine)thieno[3,2-d]pyrimidine-2-substituted)piperazine-1-substituted)(pyridine-2-substituted ) methyl ketone;

N-(5-methyl-1H-pyrazole-3-substituted)-2-(4-(4-methylpiperazine-1-substituted)piperidine-1-substituted)thieno[3,2-d ]pyrimidin-4-amine;

N-(5-methyl-1H-pyrazole-3-substituted)-4-morpholinylthieno[3,2-d]pyrimidin-2-amine;

Ethyl 4-(4-(2-(5-fluoro-2-methylaniline)thieno[3,2-d]pyrimidine-4-substituted)piperazine-1-substituted)benzoate;

N ⁴ -methyl-N ² -(5-methyl-1H-pyrazole-3-substituted)-N ⁴ -phenylthieno[3,2-d]pyrimidine-2,4-diamine;

N ² , N ⁴ -dimethyl-N ² , N ⁴ -diphenylthieno[3,2-d]pyrimidine-2,4-diamine;

4,4'-(thieno[3,2-d]pyrimidine-2,4-disubstituted)dimorpholine;

(4-(4-(4-morpholinylanilino)thieno[3,2-d]pyrimidine-2-substituted)piperazine-1-substituted)ethyl benzoate;

Ethyl 4-(4-(4-(4-morpholinylanilino)thieno[3,2-d]pyrimidine-2-substituted)piperazine-1-substituted)benzoate;

N ² -(3,4-dimethoxyphenyl)-N ⁴ -(4-morpholinylphenyl)thieno[3,2-d]pyrimidine-2,4-diamine;

N ² -(4-fluoro-2-methylphenyl)-N ⁴ -(4-morpholinylphenyl)thieno[3,2-d]pyrimidine-2,4-diamine;

N-(3,4-dimethoxyphenyl)-4-morpholinylthieno[3,2-d]pyrimidin-2-amine;

N ⁴ -(4-morpholinylphenyl)-N ² -(pyridine-2-substituted methyl)thieno[3,2-d]pyrimidine-2,4-diamine;

N-methyl-4-morpholinyl-N-phenylthieno[3,2-d]pyrimidin-2-amine;

4-(4-(cyclopropylmethyl)piperazine-1-substituted)-N-(5-methyl-1H-pyrazole-3-substituted)thieno[3,2-d]pyrimidine-2- amine;

4-(4-(2-(5-Methyl-1H-pyrazole-3-substituted amino)thieno[3,2-d]pyrimidine-4-substituted)piperazine-1-substituted)ethyl benzoate ester;

N ⁴ -(4-fluoro-2-methylphenyl)-N ² -(pyridine-3-substituted methyl)thieno[3,2-d]pyrimidine-2,4-diamine;

N ⁴ -(4-fluoro-2-methylphenyl)-N ² -methyl-N ² -phenylthieno[3,2-d]pyrimidine-2,4-diamine;

2-(2,6-dimethoxyphenyl)-N-(5-methyl-1H-pyrazole-3-substituted)thieno[3,2-d]pyrimidin-4-amine;

N ² , N ⁴ -bis(4-fluoro-2-methylphenyl)thieno[3,2-d]pyrimidine-2,4-diamine;

N ⁴ -phenyl-N ² -(5-fluoro-2-methylphenyl)thieno[3,2-d]pyrimidine-2,4-diamine;

N ⁴ -(5-methyl-1H-pyrazole-3-substituted)-N ² -(3-methylbenzyl)thieno[3,2-d]pyrimidine-2,4-diamine hydrochloride ;

(4-(4-(5-methyl-1H-pyrazole-3-substituted amine)thieno[3,2-d]pyrimidine-2-substituted)piperazine-1-substituted)(pyridine-4-substituted ) ketone hydrochloride;

2-(4-fluorophenyl)-1-(4-(4-(5-methyl-1H-pyrazole-3-substituted amine)thieno[3,2-d]pyrimidine-2-substituted)piper pyridine-1-substituted) ethyl ketone mesylate;

N ² -(3-bromobenzyl)-N ⁴ -(5-methyl-1H-pyrazole-3-substituted)thieno[3,2-d]pyrimidine-2,4-diamine methanesulfonate .

The present invention also provides a pharmaceutical composition for treating tumors, which is composed of the above-mentioned thieno-2,4-substituted pyrimidine compound or its pharmaceutically acceptable salt or stereoisomer or prodrug molecule and pharmaceutical acceptable carrier composition.

The above-mentioned thieno-2,4-substituted pyrimidine compounds or pharmaceutically acceptable salts or stereoisomers or prodrug molecules are used in the preparation of targeted anti-tumor drugs, prevention and/or treatment of cancer, infection, inflammation and autoimmune diseases Use in pharmaceutical compositions.

The diseases mentioned include: viral infections (such as HIV and Kaposi's sarcoma); inflammatory and autoimmune diseases (such as colitis, arthritis, Alzheimer's disease, glomerulonephritis, and wound healing); bacterial, Fungal and or parasitic infections; leukemias, lymphomas, and solid tumors (e.g., carcinomas and sarcomas); skin disorders (e.g., psoriasis); proliferative disorders characterized by increased cell numbers (e.g., fibroblasts, hepatocytes, bone and marrow cells, cartilage or smooth muscle cells or epithelial cells (such as endometrial hyperplasia)); bone disease and cardiovascular disease (such as restenosis and hypertrophy).

The above-mentioned thieno-2,4-substituted pyrimidine compounds or pharmaceutically acceptable salts or stereoisomers or prodrug molecules can effectively inhibit the abnormal expression of Aurora kinase, and have special inhibitory effects on Aurora-A and Aurora-B, In particular, it can significantly inhibit human cervical tumor cells, human macrophage leukemia cells, human T lymphocyte cell lines, human fibrosarcoma cells, human skin basal cell carcinoma cells, non-small cell lung cancer cells, and human breast ductal tumor cells , the proliferation of breast cancer cells, etc., can be applied to the new field of molecular targeted therapy or used to prepare drugs for treating hyperproliferative diseases.

Detailed ways

The invention will be further elaborated below in conjunction with specific embodiments.

In addition to standard methods known in the literature or exemplified in the experimental procedures, the reactions shown in the schemes of the Examples below can be employed to prepare compounds of the invention. Accordingly, the illustrative schemes that follow are for purposes of illustration and are not limiting to the compounds listed or to any particular substituents. The number of substituents shown in the schemes does not necessarily correspond to the number used in the claims, and for the sake of clarity, it is shown that the single substituent is connected to the allowed under the definition of formula (I), (II), (III), (IV) on compounds with multiple substituents.

The abbreviations in the following examples represent the following reagents or reaction conditions:

(1) DCM: dichloromethane; (2) DMF: N,N-dimethylformamide; (3) TFA: trifluoroacetic acid; (4) HATU: 2-(7-azobenzotriazole )-N, N, N', N'-tetramethylurea hexafluorophosphate; (5) DIPEA: N, N-diisopropylethylamine; (6) POCl ₃ : phosphorus oxychloride; (7 )rt: room temperature.

The NMR spectrometer and mass spectrometer used are as follows:

(1), nuclear magnetic resonance spectrometer model: VANCEAV400MHz, manufacturer: Switzerland Bruker company; Solvent is deuterated dimethyl sulfoxide-d ₆ , if use other solvents, there will be clearly mentioned in the embodiment; Chemical shift is with Standard tetramethylsilane (δ=0.00ppm) for reference;

The multiplicity of peaks is indicated as follows: s: singlet; d: doublet; dd: two doublets; t: triplet; q: quartet; qu: quintet; m: multiplet; brs: broad singlet peak.

(2) Model of mass spectrometer: Agilent 1200/MSD, manufacturer: Agilent; ESI: electronic spray ionization.

Example 1N ^4- (5-methyl-1H-pyrazole-3-substituted)-N ^2- (3-methylbenzyl)thieno[3,2-d]pyrimidine-2,4-diamine Preparation (LD4484)

Synthetic route of N ⁴ -(5-methyl-1H-pyrazole-3-substituted)-N ² -(3-methylbenzyl)thieno[3,2-d]pyrimidine-2,4-diamine as follows:

Specifically include the following steps:

(1), the synthesis of thieno[3,2-d]pyrimidine-2,4-dione, its chemical structural formula is: The synthesis steps are:

Add methyl 3-aminopyrimidine-2-carboxylate (50 g, 0.318 mol) and urea (110 g, 1.96 mol) into a round bottom flask, heat to 150° C. and reflux for 8 hours. Cool down to 90°C, add water and stir overnight, filter and dry in vacuo to obtain a white solid, which is thieno[3,2-d]pyrimidine-2,4-dione (50.2 g, yield: 94%).

The characterization data of this compound are:

¹ H NMR (400 MHz, d-DMSO), δ 11.22 (t, J=8Hz, 2H), 8.04 (m, J=6.8Hz, 1H), 6.90 (m, J=6.4Hz, 1H).

MS (ESI), m/z: 169 (M ⁺ +H ⁺ ).

(2), the synthesis of 2,4-dichlorothieno[3,2-d]pyrimidine, the chemical structural formula is: The synthesis steps are:

Add thieno[3,2-d]pyrimidine-2,4-dione (50.2 g, .298 mol), POCl ₃ (300 ml, 3.2 mol) into a round bottom flask, and reflux at 130°C until the reaction is complete. After the reaction solution was cooled to room temperature, the solvent was slowly poured out, and the remaining solid was washed with a small amount of dichloromethane, then ice-water mixture and ethyl acetate were added to the remaining solid, and the liquid was separated after the solid was completely dissolved, and the water was extracted with ethyl acetate. The organic phases were combined, washed with water, dried, and concentrated to give a white solid. Silica gel column chromatography separated the obtained white solid as 2,4-dichlorothieno[3,2-d]pyrimidine (54.1 g, yield: 89%).

The characterization data of this compound are:

¹ H NMR (400 MHz, d-DMSO), δ 6.90 (m, J=6.4 Hz, 1H), 8.04 (m, J=6.8 Hz, 1H).

MS (ESI), m/z: 205 (M ⁺ +H ⁺ ).

(3), the synthesis of 2-chloro-N-(5-methyl-1H-pyrazole-3-substituted) thieno[3,2-d]pyrimidin-4-amine, the chemical structural formula is: The synthesis steps are:

2,4-Dichlorothieno[3,2-d]pyrimidine (10 g, 49 mmol) and 5-methyl-1H-pyrazol-3-amine (5.5 g, 56.7 mmol) were dissolved in 30 mL at room temperature To the dimethylformamide, 9.8 ml of N,N-diisopropylethylamine was added, and the reaction was stirred until the reaction was complete. Pour the reaction solution into water and stir, filter out the solid, dry it, wash it with ether 3-4 times, and dry it to get a white solid to obtain 2-chloro-N-(5-methyl-1H-pyrazole-3- substituted) thieno[3,2-d]pyrimidin-4-amines (12.5 g, 96% yield).

¹ HNMR (400MHz, d-DMSO), δ12.35(s, 1H), 10.53(s, 1H), 8.20(d, J=5.6Hz, 1H), 7.34(d, J=5.2Hz, 1H), 6.34(s, 1H), 2.27(s, 3H).

MS (ESI), m/z: 265 (M ⁺ +H ⁺ ).

(4), N ⁴ -(5-methyl-1H-pyrazole-3-substituted)-N ² -(3-methylbenzyl)thieno[3,2-d]pyrimidine-2,4-di The synthesis of amine, the chemical structural formula is: The synthesis steps are:

At room temperature, 2-chloro-N-(5-methyl-1H-pyrazole-3-substituted)thieno[3,2-d]pyrimidin-4-amine (0.30g, 1.13mmol ) and o-methylphenethylamine (0.411g, 3.39mmol), the tube was sealed and reacted at 140°C until the reaction was complete. After the reaction was completed, the solvent was spin-dried, extracted with EtOAc and water, the organic phase was washed with saturated brine, dried over anhydrous Na ₂ SO ₄ , and separated by silica gel column chromatography to obtain a white solid, namely N ⁴ -(5-methyl-1H- Pyrazole-3-substituted) ^-N2- (3-methylbenzyl)thieno[3,2-d]pyrimidine-2,4-diamine (0.356 g, yield: 90%).

The characterization data of this compound are:

¹ HNMR (400MHz, d-DMSO), δ12.00(br, 1H), 9.58(br, 1H), 7.95(d, J=4Hz, 1H), 7.18(d, J=136.8Hz, 6H), 4.50 (d, J=6Hz, 2H), 2.23 (d, J=30Hz, 6H).

MS (ESI), m/z: 351 (M ⁺ +H ⁺ ).

Preparation of Example 2N ^2- (3-bromobenzyl)-N ^4- (5-methyl-1H-pyrazole-3-substituted)thieno[3,2-d]pyrimidine-2,4-diamine (LD4481)

The chemical structure of N ² -(3-bromobenzyl)-N ⁴ -(5-methyl-1H-pyrazole-3-substituted)thieno[3,2-d]pyrimidine-2,4-diamine is : Its synthesis steps are the same as in Example 1.

The characterization data of this compound are:

H ¹ NMR (400MHz, d-DMSO), δ12.26(br, 1H), 10.31(br, 1H), 8.06(s, 1H), 7.98(br, 1H), 7.56(s, 1H), 7.42( d, J=7.6Hz, 1H), 7.35(t, J=14Hz, 1H), 7.28(t, J=15.2Hz, 1H), 7.16(d, J=5.2Hz, 1H), 6.21(br, 1H ), 4.58(t, J=6Hz, 2H), 2.20(s, 3H).

MS (ESI), m/z: 416 (M ⁺ +H ⁺ ).

Example 3N ^2- (4-bromo-2-fluorobenzyl)-N ^4- (5-methyl-1H-pyrazole-3-substituted)thieno[3,2-d]pyrimidine-2,4- Preparation of Diamines (LD4478)

N ² -(4-bromo-2-fluorobenzyl)-N ⁴ -(5-methyl-1H-pyrazole-3-substituted)thieno[3,2-d]pyrimidine-2,4-diamine The chemical structural formula is Its synthesis steps are the same as in Example 1.

The characterization data of this compound are:

¹ HNMR (400MHz, d-DMSO), δ12.12(br, 1H), 9.69(br, 1H), 7.91(d, J=3.6Hz, 1H), 7.69(t, J=10.4Hz, 2H), 7.43(m, J=71.6Hz, 1H), 7.186(brs, 1H), 7.05(d, J=3.6Hz, 1H), 4.52(d, J=6Hz, 2H), 2.17(s, 3H).

MS (ESI), m/z: 434 (M ⁺ +H ⁺ ).

Example 4 Preparation of N ² -benzhydryl-N ⁴ -(5-methyl-1H-pyrazole-3-substituted)thieno[3,2-d]pyrimidine-2,4-diamine (LD4413)

The chemical structural formula of N ² -benzhydryl-N ⁴ -(5-methyl-1H-pyrazole-3-substituted)thieno[3,2-d]pyrimidine-2,4-diamine is: Its synthesis steps are the same as in Example 1.

The characterization data of this compound are:

H ¹ NMR (400MHz, d-DMSO), δ12.01(br, 1H), 9.651(br, 1H), 8.16(s, 1H), 7.93(d, J=30.4Hz, 1H), 7.39(d, J=7.2Hz, 4H), 7.31(s, 4H), 7.22(d, J=6Hz, 2H), 7.06(d, J=22.4Hz, 1H), 6.38(s, 1H), 2.20(d, J =36.4Hz, 3H).

MS (ESI), m/z: 413 (M ⁺ +H ⁺ ).

Example 5N ² -(1-benzylpiperidine-4-substituted)-N ⁴ -(5-methyl-1H-pyrazole-3-substituted)thieno[3,2-d]pyrimidine-2,4 - Preparation of diamines; (LD4430)

N ² -(1-benzylpiperidine-4-substituted)-N ⁴ -(5-methyl-1H-pyrazole-3-substituted)thieno[3,2-d]pyrimidine-2,4-di The chemical structural formula of an amine is: Its synthesis steps are the same as in Example 1.

The characterization data of this compound are:

H ¹ NMR (400MHz, d-DMSO), δ12.01(s, 1H), 9.63(d, J=41.2Hz, 1H), 7.90(t, J=32.8Hz, 1H), 7.32(t, J= 14, 5H), 7.13(m, J=105.6Hz, 1H), 6.47(d, J=62.8Hz, 1H), 3.72(d, J=2.8Hz, 1H), 3.48(s, 2H), 2.80( s, 2H), 2.23 (s, 2H), 2.14 (s, 3H), 1.88 (d, J=7.2Hz, 2H), 1.49 (d, J=10.8Hz, 2H).

MS (ESI), m/z: 420 (M ⁺ +H ⁺ ).

Example 6N ² -((2-chloropyridine-4-substituted)methyl)-N ⁴ -(5-methyl-1H-pyrazole-3-substituted)thieno[3,2-d]pyrimidine-2 , Preparation of 4-diamine (LD4443)

N ² -((2-chloropyridine-4-substituted)methyl)-N ⁴ -(5-methyl-1H-pyrazole-3-substituted)thieno[3,2-d]pyrimidine-2,4 - The chemical structural formula of diamine is: Its synthesis steps are the same as in Example 1.

The characterization data of this compound are:

H ¹ NMR (400MHz, d-DMSO), δ12.05(br, 1H), 9.62(br, 1H), 8.38(s, 1H), 7.89(s, 2H), 7.80(d, J=7.2Hz, 1H), 7.43(d, J=8Hz, 1H), 7.19(s, 1H), 7.03(s, 1H), 4.51(d, J=4.8Hz, 2H), 2.19(s, 3H).

MS (ESI), m/z: 372 (M ⁺ +H ⁺ ).

Example 7 N ² -(benzo[d][1,3]dioxolane-5-substituted methyl)-N ⁴ -(5-methyl-1H-pyrazole-3-substituted)thieno[3 ,2-d] Preparation of pyrimidine-2,4-diamine (LD4442)

N ² -(Benzo[d][1,3]dioxolane-5-substituted methyl)-N ⁴ -(5-methyl-1H-pyrazole-3-substituted)thieno[3,2 -d] The chemical structural formula of pyrimidine-2,4-diamine is: Its synthesis steps are the same as in Example 1.

The characterization data of this compound are:

H ¹ NMR (400MHz, d-DMSO), δ12.01(br, 1H), 9.59(br, 1H), 7.87(s, 1H), 7.02(s, 1H), 6.91(s, 1H), 6.76( s, 2H), 5.94 (s, 2H), 4.42 (d, J=6Hz, 2H), 2.18 (d, J=20.4Hz, 3H).

MS (ESI), m/z: 381 (M ⁺ +H ⁺ ).

Example 8 Preparation of N ² -(2-fluorobenzyl)-N ⁴ -(5-methyl-1H-pyrazole-3-substituted)thieno[3,2-d]pyrimidine-2,4-diamine (LD4445)

The chemical structural formula of N ² -(2-fluorobenzyl)-N ⁴ -(5-methyl-1H-pyrazole-3-substituted)thieno[3,2-d]pyrimidine-2,4-diamine is : Its synthesis steps are the same as in Example 1.

The characterization data of this compound are:

H ¹ NMR (400MHz, d-DMSO), δ12.08(br, 1H), 9.63(br, 1H), 7.92(d, J=30.4Hz, 1H), 7.20(m, J=133.6Hz, 6H) , 6.28 (br, 1H), 4.57 (d, J=5.6Hz, 2H), 2.16 (s, 3H).

MS (ESI), m/z: 355 (M ⁺ +H ⁺ ).

Example 9 Preparation of N ^2- (4-chlorobenzyl)-N ^4- (5-methyl-1H-pyrazole-3-substituted)thieno[3,2-d]pyrimidine-2,4-diamine (LD4476)

The chemical structure of N ² -(4-chlorobenzyl)-N ⁴ -(5-methyl-1H-pyrazole-3-substituted)thieno[3,2-d]pyrimidine-2,4-diamine is : Its synthesis steps are the same as in Example 1.

The characterization data of this compound are:

H ¹ NMR (400MHz, d-DMSO), δ12.10(br, 1H), 10.17(br, 1H), 7.94(s, 1H), 7.36(s, 5H), 7.06(s, 1H), 4.52( d, J=6.4Hz, 2H), 2.18(s, 3H).

MS (ESI), m/z: 371 (M ⁺ +H ⁺ ).

Example 10N ^2- (2-chloro-6-fluorobenzyl)-N ⁴ -(5-methyl-1H-pyrazole-3-substituted)thieno[3,2-d]pyrimidine-2,4- Preparation of Diamines (LD4483)

N ² -(2-chloro-6-fluorobenzyl)-N ⁴ -(5-methyl-1H-pyrazole-3-substituted)thieno[3,2-d]pyrimidine-2,4-diamine The chemical structure formula is: Its synthesis steps are the same as in Example 1.

The characterization data of this compound are:

H ¹ NMR (400MHz, d-DMSO), δ12.29(d, J=221.2Hz, 1H), 9.97(d, J=269.2Hz, 1H), 7.95(d, J=43.6Hz, 1H), 7.27 (m, J=162Hz, 5H), 6.68(d, J=51.6Hz, 1H), 4.63(d, J=3.6Hz, 2H), 2.13(d, J=32.8Hz3H).

MS (ESI), m/z: 389 (M ⁺ +H ⁺ ).

Example 11N ² -(2,6-dichlorobenzyl)-N ⁴ -(5-methyl-1H-pyrazole-3-substituted)thieno[3,2-d]pyrimidine-2,4-di Preparation of Amines (LD4479)

N ² -(2,6-dichlorobenzyl)-N ⁴ -(5-methyl-1H-pyrazole-3-substituted)thieno[3,2-d]pyrimidine-2,4-diamine The chemical structural formula is: Its synthesis steps are the same as in Example 1.

The characterization data of this compound are:

¹ HNMR (400MHz, d-DMSO), δ12.29(d, J=222.8Hz, 1H), 9.99(d, J=272.8Hz, 1H), 7.96(d, J=40Hz, 1H), 7.44(d , J=47.6Hz, 4H), 7.15(d, J=41.6Hz, 1H), 6.67(s, 1H), 4.72(d, J=4.4Hz, 2H), 2.17(d, J=28.8Hz, 3H ).

MS(ESI), m/z: 406(M ⁺ +H ⁺ ).

Example 12N ⁴ -(5-methyl-1H-pyrazole-3-substituted)-N ² -(4-(trifluoromethyl)benzyl)thieno[3,2-d]pyrimidine-2,4 - Preparation of diamines (LD4485)

N ⁴ -(5-methyl-1H-pyrazole-3-substituted)-N ² -(4-(trifluoromethyl)benzyl)thieno[3,2-d]pyrimidine-2,4-di The chemical structural formula of an amine is: Its synthesis steps are the same as in Example 1.

The characterization data of this compound are:

¹ HNMR (400MHz, d-DMSO), δ12.03(s, 1H), 9.64(s, 1H), 7.89(s, 1H), 7.66(d, J=7.6Hz, 2H), 7.56(s, 2H ), 7.25(s, 1H), 7.02(s, 1H), 4.75(s, 2H), 2.16(s, 3H).

MS (ESI), m/z: 405 (M ⁺ +H ⁺ ).

Example 13N ^2- (4-chlorophenethyl)-N ^4- (5-methyl-1H-pyrazole-3-substituted)thieno[3,2-d]pyrimidine-2,4-diamine Preparation (LD4480)

The chemical structure of N ² -(4-chlorophenethyl)-N ⁴ -(5-methyl-1H-pyrazole-3-substituted)thieno[3,2-d]pyrimidine-2,4-diamine for: Its synthesis steps are the same as in Example 1.

The characterization data of this compound are:

¹ HNMR (400MHz, d-DMSO), δ12.07(br, 1H), 9.60(br, 1H), 7.94(d, J=32Hz, 1H), 7.34(m, J=46.4Hz, 4H), 7.03 (s, 1H), 6.62 (s, 1H), 6.51 (s, 1H), 3.51 (s, 2H), 2.87 (m, J=20.4Hz, 2H), 2.21 (s, 3H).

MS (ESI), m/z: 386 (M ⁺ +H ⁺ ).

Example 14 N ² -(2,4-difluorophenyl)-N ⁴ -(5-methyl-1H-pyrazole-3-substituted)thieno[3,2-d]pyrimidine-2,4-di Preparation of Amines (LD4473)

N ² -(2,4-difluorophenyl)-N ⁴ -(5-methyl-1H-pyrazole-3-substituted)thieno[3,2-d]pyrimidine-2,4-diamine The chemical structural formula is: Its synthesis steps are the same as in Example 1.

The characterization data of this compound are:

¹ HNMR (400MHz, d-DMSO), δ12.43(br, 1H), 10.18(br, 1H), 8.46(s, 1H), 8.16(s, 1H), 7.79(s, 1H), 7.28(m , J=22.4Hz, 1H), 7.14(s, 1H), 7.06(t, J=16.4Hz, 1H), 6.22(m, J=83.6Hz, 1H), 2.17(s, 3H).

MS (ESI), m/z: 359 (M ⁺ +H ⁺ ).

Example 15 Preparation of N ^2- (3-bromophenyl)-N ^4- (5-methyl-1H-pyrazole-3-substituted)thieno[3,2-d]pyrimidine-2,4-diamine (LD4420)

The chemical structure of N ² -(3-bromophenyl)-N ⁴ -(5-methyl-1H-pyrazole-3-substituted)thieno[3,2-d]pyrimidine-2,4-diamine is : Its synthesis steps are the same as in Example 1.

The characterization data of this compound are:

¹ HNMR (400MHz, d-DMSO), δ12.15(br, 1H), 9.85(s, 1H), 9.26(s, 1H), 8.15(d, J=23.24Hz, 1H), 8.02(d, J =5.2Hz, 1H), 7.77(d, J=7.2Hz, 1H), 7.23(m, J=59.6Hz, 2H), 7.05(t, J=23.6Hz, 1H), 6.51(s, 1H), 2.27(s, 3H).

MS (ESI), m/z: 402 (M ⁺ +H ⁺ ).

Example 16N ^4- (5-methyl-1H-pyrazole-3-substituted)-N ² -(3-morpholinopropyl)thieno[3,2-d]pyrimidine-2,4-diamine Preparation of (LD4419)

Chemistry of N ⁴ -(5-methyl-1H-pyrazole-3-substituted)-N ² -(3-morpholinopropyl)thieno[3,2-d]pyrimidine-2,4-diamine The structural formula is: Its synthesis steps are the same as in Example 1.

The characterization data of this compound are:

¹ HNMR (400MHz, d-DMSO), δ12.03(br, 1H), 9.75(br, 1H), 7.84(t, J=22.8Hz, 1H), 7.03(m, J=20.4Hz, 1H), 6.58(s, 1H), 3.57(t, J=8.8Hz, 4H), 3.30(s, 2H), 2.23(s, 6H), 2.21(s, 3H), 1.71(m, J=27.6Hz, 2H ).

MS (ESI), m/z: 374 (M ⁺ +H ⁺ ).

Example 17 N ² -(biphenyl-4-substituted)-N ⁴ -(5-methyl-1H-pyrazole-3-substituted)thieno[3,2-d]pyrimidine-2,4-diamine Preparation of (LD4407)

Chemistry of N ² -(biphenyl-4-substituted)-N ^4- (5-methyl-1H-pyrazole-3-substituted)thieno[3,2-d]pyrimidine-2,4-diamine The structural formula is Its synthesis steps are the same as in Example 1.

The characterization data of this compound are:

¹ HNMR (400MHz, d-DMSO), δ9.78(br, 1H), 7.89(br, 1H), 7.58(t, J=52Hz, 5H), 7.34(t, J=31.2Hz, 5H), 7.25 (m, J=21.2Hz, 2H), 6.56(br, 4H), 2.21(s, 3H).

MS (ESI), m/z: 399 (M ⁺ +H ⁺ ).

Example 18 Preparation of N-(5-methyl-1H-pyrazole-3-substituted)-2-(3-methylpiperazine-1-substituted)thieno[3,2-d]pyrimidin-4-amine (LD4421)

The chemical structure of N-(5-methyl-1H-pyrazole-3-substituted)-2-(3-methylpiperazine-1-substituted)thieno[3,2-d]pyrimidin-4-amine is Its synthesis steps are the same as in Example 1.

The characterization data of this compound are:

¹ HNMR (400MHz, d-DMSO), δ12.09(s, 1H), 9.63(s, 1H), 7.90(d, J=33.2Hz, 1H), 7.06(d, J=4.8Hz, 1H), 6.314(s, 1H), 4.47(t, J=20Hz, 2H), 4.1(d, J=4.4Hz, 1H), 3.16(d, J=4.4Hz, 2H), 2.90(d, J=10Hz, 1H), 2.76(t, J=24Hz, 1H), 2.63(t, J=20Hz, 2H), 2.40(t, J=22.4Hz, 1H), 2.24(s, 3H), 1.01(m, J= 6Hz, 3H).

MS (ESI), m/z: 330 (M ⁺ +H ⁺ ).

Example 19N ^2- (4-chloro-3-(trifluoromethyl)phenyl)-N ⁴ -(5-methyl-1H-pyrazole-3-substituted)thieno[3,2-d]pyrimidine - Preparation of 2,4-diamine (LD4422)

N ² -(4-chloro-3-(trifluoromethyl)phenyl)-N ⁴ -(5-methyl-1H-pyrazole-3-substituted)thieno[3,2-d]pyrimidine-2 , the chemical structure of 4-diamine is Its synthesis steps are the same as in Example 1.

The characterization data of this compound are:

¹ HNMR (400MHz, d-DMSO), δ12.17(br, 1H), 9.92(br, 1H), 9.54(s, 1H), 8.35(s,

1H), 8.25(d, J=7.6Hz, 1H), 8.04(d, J=5.6Hz, 1H), 7.55(d, J=8.8Hz, 1H), 7.20(d, J=5.2Hz, 1H) , 6.54 (d, J=22Hz, 1H), 2.26 (s, 3H).

MS (ESI), m/z: 425 (M ⁺ +H ⁺ ).

Example 20N ^2- (4-bromo-3-fluorophenyl)-N ^4- (5-methyl-1H-pyrazole-3-substituted)thieno[3,2-d]pyrimidine-2,4- Preparation of Diamines (LD4424)

N ² -(4-bromo-3-fluorophenyl)-N ⁴ -(5-methyl-1H-pyrazole-3-substituted)thieno[3,2-d]pyrimidine-2,4-diamine The chemical structural formula is Its synthesis steps are the same as in Example 1.

The characterization data of this compound are:

¹ HNMR (400MHz, d-DMSO), δ12.20(br, 1H), 9.87(s, 1H), 9.46(br, 1H), 8.19(d, J=12.8Hz, 1H), 8.04(s, 1H) ), 7.36(s, 2H), 7.00(s, 1H), 2.27(s, 3H).

MS (ESI), m/z: 420 (M ⁺ +H ⁺ ).

Example 21N ^2- (3-fluoro-5-(trifluoromethyl)phenyl)-N ⁴ -(5-methyl-1H-pyrazole-3-substituted)thieno[3,2-d]pyrimidine - Preparation of 2,4-diamine (LD4432)

N ² -(3-fluoro-5-(trifluoromethyl)phenyl)-N ⁴ -(5-methyl-1H-pyrazole-3-substituted)thieno[3,2-d]pyrimidine-2 , the chemical structure of 4-diamine is Its synthesis steps are the same as in Example 1.

The characterization data of this compound are:

¹ HNMR (400MHz, d-DMSO), δ12.20(br, 1H), 9.65(br, 1H), 9.68(s, 1H), 8.32(d, J=12Hz, 1H), 8.06(d, J= 4.8Hz, 1H), 7.23(d, J=5.2Hz, 1H), 7.04(d, J=8Hz, 1H), 6.48(s, 1H), 2.27(s, 3H).

MS (ESI), m/z: 409 (M ⁺ +H ⁺ ).

Example 22N ² -(2,6-diethylphenyl)-N ⁴ -(5-methyl-1H-pyrazole-3-substituted)thieno[3,2-d]pyrimidine-2,4- Preparation of Diamines (LD4437)

N ² -(2,6-diethylphenyl)-N ⁴ -(5-methyl-1H-pyrazole-3-substituted)thieno[3,2-d]pyrimidine-2,4-diamine The chemical structural formula is Its synthesis steps are the same as in Example 1.

The characterization data of this compound are:

¹ HNMR (400MHz, d-DMSO), δ11.80(br, 1H), 9.69(br, 1H), 8.08(s, 1H), 8.66(s, 1H), 7.18(d, J=6Hz, 1H) , 7.13(d, J=6.8Hz, 2H), 7.03(s, 1H), 2.52(t, J=19.6Hz, 4H), 2.04(d, J=9.6Hz, 3H), 1.06(m, J= 24.4Hz, 6H).

MS (ESI), m/z: 379 (M ⁺ +H ⁺ ).

Example 23 Preparation of N ² -methyl-N ⁴ -(5-methyl-1H-pyrazole-3-substituted)-N2-phenylthieno[3,2-d]pyrimidine-2,4-diamine (LD1140)

The chemical structure of N ² -methyl-N ⁴ -(5-methyl-1H-pyrazole-3-substituted)-N2-phenylthieno[3,2-d]pyrimidine-2,4-diamine is Its synthesis steps are the same as in Example 1.

The characterization data of this compound are:

¹ HNMR (400MHz, d-DMSO), δ11.87(s, 1H), 9.80(s, 1H), 7.95(d, J=4Hz, 1H), 7.42(t, J=14.8Hz, 2H), 7.33 (d, J=7.6Hz, 2H), 7.24(t, J=13.6Hz, 1H), 7.14(d, J=4.4Hz, 1H), 5.75(s, 1H), 3.7(s, 3H), 2.04 (s, 3H).

MS (ESI), m/z: 337 (M ⁺ +H ⁺ ).

Example 24 N ² -(5-fluoro-2-methylphenyl)-N ⁴ -(5-methyl-1H-pyrazole-3-substituted)thieno[3,2-d]pyrimidine-2,4 - Preparation of diamines (LD1148)

N ² -(5-fluoro-2-methylphenyl)-N ⁴ -(5-methyl-1H-pyrazole-3-substituted)thieno[3,2-d]pyrimidine-2,4-di The chemical structure of amine is Its synthesis steps are the same as in Example 1.

The characterization data of this compound are:

¹ HNMR (400MHz, d-DMSO), δ12.08(br, 1H), 9.87(br, 1H), 8.14(s, 1H), 8.02(s, 1H), 7.73(s, 1H), 7.19(t , J=15.2Hz, 2H), 6.80(m, J=16.4Hz, 1H), 6.30(br, 1H), 2.25(s, 3H), 2.18(s, 3H).

MS (ESI), m/z: 355 (M ⁺ +H ⁺ ).

Example 25 N ² -(3,5-bis(trifluoromethyl)benzyl)-N ⁴ -(5-methyl-1H-pyrazole-3-substituted)thieno[3,2-d]pyrimidine- Preparation of 2,4-diamine (LD4486)

N ² -(3,5-bis(trifluoromethyl)benzyl)-N ⁴ -(5-methyl-1H-pyrazole-3-substituted)thieno[3,2-d]pyrimidine-2, The chemical structural formula of 4-diamine is Its synthesis steps are the same as in Example 1.

The characterization data of this compound are:

¹ HNMR (400MHz, d-DMSO), δ12.03(br, 1H), 9.75(br, 1H), 7.96(t, J=56.4Hz, 4H), 7.39(d, J=52.4Hz, 1H), 7.01(s, 1H), 6.28(m, J=130Hz, 1H), 4.67(s, 1H), 2.17(s, 3H).

MS (ESI), m/z: 473 (M ⁺ +H ⁺ ).

Example 26 4-(4-(2-(5-methyl-1H-pyrazole-3-substituted amino)thieno[3,2-d]pyrimidine-4-substituted)piperazine-1-substituted)benzene Preparation of ethyl formate (LD1192)

4-(4-(2-(5-Methyl-1H-pyrazole-3-substituted amino)thieno[3,2-d]pyrimidine-4-substituted)piperazine-1-substituted)ethyl benzoate The chemical structure of ester is Its synthesis steps are the same as in Example 1.

The characterization data of this compound are:

¹ HNMR (400MHz, d-DMSO), δ12.09(s, 1H), 9.74(s, 1H), 7.94(d, J=5.6Hz, 1H), 7.8(d, J=8.8Hz, 2H), 7.10(d, J=5.6Hz, 1H), 7.04(d, J=8.8Hz, 2H), 6.38(s, 1H), 4.23(m, J=26Hz, 2H), 3.87(t, J=10Hz, 4H), 3.43(t, J=10Hz, 4H), 2.26(s, 3H), 1.29(t, J=14Hz, 3H).

MS (ESI), m/z: 464 (M ⁺ +H ⁺ ).

Example 27 2-(4-methyl-1,4-diazepane-1-substituted)-N-(5-methyl-1H-pyrazole-3-substituted)thieno[3,2- d] Preparation of pyrimidin-4-amine (LD4415)

2-(4-methyl-1,4-diazepane-1-substituted)-N-(5-methyl-1H-pyrazole-3-substituted)thieno[3,2-d] The chemical structural formula of pyrimidin-4-amine is Its synthesis steps are the same as in Example 1.

¹ HNMR (400MHz, d-DMSO), δ12.05(s, 1H), 9.64(s, 1H), 7.88(d, J=5.2Hz, 1H), 7.05(d, J=5.2Hz, 1H), 6.37(s, 1H), 3.84(d, J=4Hz, 2H), 3.76(t, J=12Hz, 2H), 2.62(d, J=4.4Hz, 2H), 2.46(d, J=5.2Hz, 2H), 2.24(d, J=8.4Hz, 6H), 1.89(d, J=5.2Hz, 2H).

MS (ESI), m/z: 344 (M ⁺ +H ⁺ ).

Example 28 N ² , Preparation of N ⁴ -bis(5-methyl-1H-pyrazole-3-substituted)thieno[3,2-d]pyrimidine-2,4-diamine (LD4428)

The chemical structure of N ² , N ⁴ -bis(5-methyl-1H-pyrazole-3-substituted)thieno[3,2-d]pyrimidine-2,4-diamine is Its synthesis steps are the same as in Example 1.

The characterization data of this compound are:

¹ HNMR (400MHz, d-DMSO), δ11.80(br, 1H), 10.36(br, 1H), 9.81(br, 1H), 8.89(br, 1H), 8.06(s, 1H), 7.19(s , 1H), 5.92 (br, 1H), 5.31 (s, 1H), 2.19 (d, J=34Hz, 6H).

MS (ESI), m/z: 327 (M ⁺ +H ⁺ ).

Example 29 N-(5-methyl-1H-pyrazole-3-substituted)-2-(4-(4-methylpiperazine-1-substituted)piperidine-1-substituted)thieno[3,2 -d] Preparation of pyrimidin-4-amine (LD4487)

N-(5-methyl-1H-pyrazole-3-substituted)-2-(4-(4-methylpiperazine-1-substituted)piperidine-1-substituted)thieno[3,2-d ] The chemical structural formula of pyrimidin-4-amine is Its synthesis steps are the same as in Example 1.

The characterization data of this compound are:

¹ HNMR (400MHz, d-DMSO), δ12.09(s, 1H), 9.62(s, 1H), 7.90(d, J=5.2Hz, 1H), 7.05(d, J=5.2Hz, 1H), 6.31(s, 1H), 4.67(d, J=12.8Hz, 2H), 3.35(m, J=21.6Hz, 2H), 3.16(s, 1H), 2.82(t, J=23.6Hz, 2H), 2.41(m, J=11.2Hz, 4H), 2.24(s, 3H), 2.13(s, 3H), 1.8(d, J=10.8Hz, 2H), 1.31(m, J=23.2Hz, 2H).

MS (ESI), m/z: 413 (M ⁺ +H ⁺ ).

Example 30 Preparation of N-(5-methyl-1H-pyrazole-3-substituted)-4-morpholinylthieno[3,2-d]pyrimidin-2-amine (LD1142)

The chemical structural formula of N-(5-methyl-1H-pyrazole-3-substituted)-4-morpholinylthieno[3,2-d]pyrimidin-2-amine is: Its synthesis steps are the same as in Example 1.

¹ HNMR (400MHz, d-DMSO), δ11.83(s, 1H), 8.77(s, 1H), 8.04(s, 1H), 7.18(s, 1H), 3.85(d, J=4.4Hz, 4H ), 3.75 (d, J=4.8Hz, 4H), 2.19 (s, 3H).

MS (ESI), m/z: 317 (M ⁺ +H ⁺ ).

Preparation of Example 314-(4-(2-(5-fluoro-2-methylaniline)thieno[3,2-d]pyrimidine-4-substituted)piperazine-1-substituted)ethyl benzoate ( LD1193)

The chemical structural formula of ethyl 4-(4-(2-(5-fluoro-2-methylaniline)thieno[3,2-d]pyrimidine-4-substituted)piperazine-1-substituted)benzoate is: Its synthesis steps are the same as in Example 1.

The characterization data of this compound are:

¹ HNMR (400MHz, CDCl3), δ8.19(m, J=14.8Hz, 1H), 7.97(d, J=9.2Hz, 2H), 7.69(d, J=5.6Hz, 1H), 7.09(t, J=15.2Hz, 1H), 6.89(d, J=8.8Hz, 2H), 6.70(s, 1H), 6.64(m, J=19.2Hz, 2H), 4.34(m, J=21.2Hz, 2H) , 4.15(m, J=20.4Hz, 4H), 3.54(m, J=10.4Hz, 4H), 2.32(t, J=14.8Hz, 3H), 1.25(t, J=14Hz, 3H).

MS (ESI), m/z: 492 (M ⁺ +H ⁺ ).

Example 32N ⁴ -methyl-N ² -(5-methyl-1H-pyrazole-3-substituted)-N ⁴ -phenylthieno[3,2-d]pyrimidine-2,4-diamine Preparation (LD1145)

The chemical structure of N ⁴ -methyl-N ² -(5-methyl-1H-pyrazole-3-substituted)-N ⁴ -phenylthieno[3,2-d]pyrimidine-2,4-diamine for: Its synthesis steps are the same as in Example 1.

The characterization data of this compound are:

¹ HNMR (500MHz, d-DMSO), δ11.73(s, 1H), 8.08(s, 1H), 7.73(s, 1H), 7.52(m, J=35.6Hz, 3H), 7.45(s, 1H ), 7.43 (s, 1H), 3.53 (s, 3H), 2.20 (d, J=4.8Hz, 3H).

MS (ESI), m/z: 337 (M ⁺ +H ⁺ ).

Example 33 N ² , N ⁴ -Dimethyl-N ² , N ⁴ -Diphenylthieno[3,2-d]pyrimidine-2,4-diamine Preparation (LD1133)

The chemical structural formula of N ² , N ⁴ -dimethyl-N ² , N ⁴ -diphenylthieno[3,2-d]pyrimidine-2,4-diamine is: Its synthesis steps are the same as in Example 1.

The characterization data of this compound are:

¹ HNMR (400MHz, d-DMSO), δ7.70(d, J=5.2Hz, 1H), 7.42(m, J=54.0Hz, 8H), 7.14(m, J=14.0Hz, 1H), 6.99( d, J=5.2Hz, 1H), 3.53(s, 3H), 3.32(s, 3H).

MS (ESI), m/z: 347 (M ⁺ +H ⁺ ).

Preparation of Example 344,4'-(thieno[3,2-d]pyrimidine-2,4-disubstituted)dimorpholine (LD1134)

The chemical structural formula of 4,4'-(thieno[3,2-d]pyrimidine-2,4-disubstituted)dimorpholine is: Its synthesis steps are the same as in Example 1.

The characterization data of this compound are:

¹ HNMR (400MHz, CDCl ₃ ), δ7.61(d, J=5.2Hz, 1H), 3.94(d, J=4.4Hz, 4H), 3.86(m, J=29.2Hz, 4H), 3.79(s , 8H).

MS (ESI), m/z: 307 (M ⁺ +H ⁺ ).

Example 35 Preparation of (4-(4-(4-morpholinylanilino)thieno[3,2-d]pyrimidine-2-substituted)piperazine-1-substituted) ethyl benzoate (LD1179)

The chemical structure of ethyl (4-(4-(4-morpholinylanilino)thieno[3,2-d]pyrimidine-2-substituted)piperazine-1-substituted)benzoate is: Its synthesis steps are the same as in Example 1.

¹ HNMR (400MHz, d-DMSO), δ8.03(d, J=5.6Hz, 1H), 7.80(d, J=8.8Hz, 2H), 7.16(d, J=5.6Hz, 1H), 7.02( d, J=9.2Hz, 2H), 4.25(m, J=24Hz, 2H), 3.84(d, J=3.6Hz, 8H), 3.74(t, J=9.2Hz, 4H), 3.41(t, J =10Hz, 4H), 1.29(t, J=14.4Hz, 3H).

MS (ESI), m/z: 454 (M ⁺ +H ⁺ ).

Example 36 Preparation of ethyl benzoate (4-(4-(4-morpholinylanilino)thieno[3,2-d]pyrimidine-2-substituted)piperazine-1-substituted)benzoate (LD1178)

The chemical structural formula of ethyl 4-(4-(4-(4-morpholinylanilino)thieno[3,2-d]pyrimidine-2-substituted)piperazine-1-substituted)benzoate is: Its synthesis steps are the same as in Example 1.

¹ HNMR (400MHz, d-DMSO), δ9.22(s, 1H), 7.96(d, J=5.2Hz, 1H), 7.80(d, J=9.2Hz, 2H), 7.55(d, J=9.2 Hz, 2H), 7.12(d, J=5.2Hz, 1H), 7.03(d, J=9.2Hz, 2H), 6.96(d, J=9.2Hz, 2H), 4.24(m, J=21.2Hz, 2H), 3.84(t, J=10Hz, 4H), 3.75(t, J=9.6Hz, 4H), 3.41(t, J=9.6Hz, 4H), 3.10(t, J=9.6Hz, 4H), 1.29(t, J=14Hz, 3H).

MS (ESI), m/z: 545 (M ⁺ +H ⁺ ).

Example 37N ^2- (3,4-dimethoxyphenyl)-N ^4- (4-morpholinylphenyl)thieno[3,2-d]pyrimidine-2,4-diamine preparation ( LD1175)

The chemical structural formula of N ² -(3,4-dimethoxyphenyl)-N ⁴ -(4-morpholinylphenyl)thieno[3,2-d]pyrimidine-2,4-diamine is: Its synthesis steps are the same as in Example 1.

The characterization data of this compound are:

¹ HNMR (400MHz, d-DMSO), δ9.20(s, 1H), 8.79(s, 1H), 7.99(d, J=5.2Hz, 1H), 7.58(d, J=8Hz, 2H), 7.47 (d, J=2Hz, 1H), 7.31(t, J=8.8Hz, 1H), 7.15(d, J=5.6Hz, 1H), 6.93(d, J=9.2Hz, 2H), 6.80(d, J=8.8Hz, 1H), 3.76(t, J=9.6Hz, 4H), 3.70(s, 3H), 3.63(s, 3H).

MS (ESI), m/z: 464 (M ⁺ +H ⁺ ).

Example 38N ^2- (4-fluoro-2-methylphenyl)-N ^4- (4-morpholinylphenyl)thieno[3,2-d]pyrimidine-2,4-diamine preparation ( LD1167)

The chemical structural formula of N ² -(4-fluoro-2-methylphenyl)-N ⁴ -(4-morpholinylphenyl)thieno[3,2-d]pyrimidine-2,4-diamine is: Its synthesis steps are the same as in Example 1.

The characterization data of this compound are:

¹ HNMR (400MHz, d-DMSO), δ10.76(brs, 1H), 9.73(brs, 1H), 8.28(brs, 1H), 7.5(m, J=8.4Hz, 1H), 7.43(d, J =6.8Hz, 2H), 7.31(t, J=14.8Hz, 2H), 7.03(d, J=6.4Hz1H), 6.91(d, J=7.2Hz, 2H), 3.75(t, J=8.4Hz, 4H), 3.11(s, 4H), 2.25(s, 3H).

MS (ESI), m/z: 436 (M ⁺ +H ⁺ ).

Example 39 Preparation of N-(3,4-dimethoxyphenyl)-4-morpholinothieno[3,2-d]pyrimidin-2-amine (LD1174)

The chemical structural formula of N-(3,4-dimethoxyphenyl)-4-morpholinylthieno[3,2-d]pyrimidin-2-amine is: Its synthesis steps are the same as in Example 1.

The characterization data of this compound are: ¹ HNMR (400MHz, d-DMSO), δ8.97(s, 1H), 8.08(d, J=5.2Hz, 1H), 7.50(s, 1H), 7.19(d, J =6Hz, 2H), 6.85(d, J=8.4Hz, 1H), 3.88(s, 4H), 3.73(t, J=21.6Hz, 14H).

MS (ESI), m/z: 373 (M ⁺ +H ⁺ ).

Example 40N ^4- (4-morpholinylphenyl)-N ² -(pyridine-2-substituted methyl)thieno[3,2-d]pyrimidine-2,4-diamine preparation (LD1182)

The chemical structural formula of N ⁴ -(4-morpholinylphenyl)-N ² -(pyridine-2-substituted methyl)thieno[3,2-d]pyrimidine-2,4-diamine is: Its synthesis steps are the same as in Example 1.

The characterization data of this compound are:

¹ HNMR (400MHz, d-DMSO), δ9.05(s, 1H), 8.51(d, J=4.4Hz, 1H), 7.90(d, J=5.6Hz, 1H), 7.69(m, J=17.2 Hz, 1H), 7.51(s, 2H), 7.31(d, J=8Hz, 1H), 7.21(m, J=12Hz, 1H), 7.05(t, J=18.4Hz, 1H), 6.84(d, J=6.4Hz, 2H), 4.58(d, J=6.4Hz, 2H), 3.74(t, J=9.2Hz, 4H), 3.06(t, J=9.2Hz, 4H).

MS (ESI), m/z: 419 (M ⁺ +H ⁺ ).

Example 41 Preparation of N-methyl-4-morpholinyl-N-phenylthieno[3,2-d]pyrimidin-2-amine (LD1147)

The chemical structural formula of N-methyl-4-morpholinyl-N-phenylthieno[3,2-d]pyrimidin-2-amine is: Its synthesis steps are the same as in Example 1.

The characterization data of this compound are:

¹ HNMR (400MHz, d-DMSO), δ8.03(d, J=5.2Hz, 1H), 7.35(m, J=10.8Hz, 4H), 7.14(m, J=20.4Hz, 2H), 3.71( t, J=5.2Hz, 4H), 3.67(d, J=4.8Hz, 4H), 3.32(s, 3H).

MS (ESI), m/z: 327 (M ⁺ +H ⁺ ).

Example 42 4-(4-(cyclopropylmethyl)piperazine-1-substituted)-N-(5-methyl-1H-pyrazole-3-substituted)thieno[3,2-d]pyrimidine- Preparation of 2-Amines (LD1186)

4-(4-(cyclopropylmethyl)piperazine-1-substituted)-N-(5-methyl-1H-pyrazole-3-substituted)thieno[3,2-d]pyrimidine-2- The chemical structural formula of an amine is: Its synthesis steps are the same as in Example 1.

The characterization data of this compound are:

¹ HNMR (400MHz, d-DMSO), δ11.72(s, 1H), 8.74(s, 1H), 8.02(d, J=4.8Hz, 1H), 7.17(s, 1H), 6.41(s, 1H ), 3.88(s, 4H), 2.57(t, J=9.2Hz, 4H), 2.32(s, 3H), 2.21(t, J=16.8Hz, 2H), 0.86(m, J=38.8Hz, 1H ), 0.47(m, J=9.2Hz, 2H), 0.08(d, J=4.64Hz, 2H).

MS (ESI), m/z: 370 (M ⁺ +H ⁺ ).

Example 434-(4-(2-(5-methyl-1H-pyrazole-3-substituted amino)thieno[3,2-d]pyrimidine-4-substituted)piperazine-1-substituted)benzene Preparation of ethyl formate (LD1190C)

4-(4-(2-(5-Methyl-1H-pyrazole-3-substituted amino)thieno[3,2-d]pyrimidine-4-substituted)piperazine-1-substituted)ethyl benzoate The chemical structural formula of an ester is: Its synthesis steps are the same as in Example 1.

The characterization data of this compound are:

¹ HNMR (400MHz, d-DMSO), δ8.26(d, J=5.6Hz, 1H), 7.82(d, J=8.4Hz, 2H), 7.46(d, J=5.6Hz, 1H), 7.00( d, J=8.8Hz, 2H), 6.66(s, 2H), 5.24(s, 1H), 4.24(m, J=20.8Hz, 2H), 4.16(s, 4H), 3.62(s, 4H), 2.08(s, 3H), 1.29(t, J=14.4Hz, 3H).

MS (ESI), m/z: 464 (M ⁺ +H ⁺ ).

Preparation of Example 44N ^4- (4-fluoro-2-methylphenyl)-N ^2- (pyridine-3-substituted methyl)thieno[3,2-d]pyrimidine-2,4-diamine ( LD46018)

The chemical structural formula of N ⁴ -(4-fluoro-2-methylphenyl)-N ² -(pyridine-3-substituted methyl)thieno[3,2-d]pyrimidine-2,4-diamine is: Its synthesis steps are the same as in Example 1.

The characterization data of this compound are:

¹ HNMR (400MHz, d-DMSO), δ8.94(s, 1H), 8.46(d, J=4Hz, 1H), 7.89(d, J=5.6Hz, 1H), 7.66(m, J=16.8Hz , 1H), 7.23(m, J=46Hz, 4H), 7.01(m, J=24.4Hz, 3H), 4.51(dJ=6Hz, 2H), 2.14(s, 3H).

MS (ESI), m/z: 366 (M ⁺ +H ⁺ ).

Example 45N ^4- (4-fluoro-2-methylphenyl)-N ² -methyl-N ² -phenylthieno[3,2-d]pyrimidine-2,4-diamine preparation (LD46017 )

The chemical structural formula of N ⁴ -(4-fluoro-2-methylphenyl)-N ² -methyl-N ² -phenylthieno[3,2-d]pyrimidine-2,4-diamine is: Its synthesis steps are the same as in Example 1.

The characterization data of this compound are:

¹ HNMR (400MHz, d-DMSO), δ8.92(s, 1H), 7.97(d, J=5.6Hz, 1H), 7.25(m, J=49.2Hz, 5H), 7.14(d, J=5.6 Hz, 1H), 7.03(d, J=53.2Hz, 1H), 6.94(m, J=16.8Hz, 1H), 3.41(s, 3H), 2.15(s, 3H).

MS (ESI), m/z: 365 (M ⁺ +H ⁺ ).

Example 46 Preparation of 2-(2,6-dimethoxyphenyl)-N-(5-methyl-1H-pyrazole-3-substituted)thieno[3,2-d]pyrimidin-4-amine (LD46016)

The chemical structure of 2-(2,6-dimethoxyphenyl)-N-(5-methyl-1H-pyrazole-3-substituted)thieno[3,2-d]pyrimidin-4-amine is : Its synthesis steps are the same as in Example 1.

The characterization data of this compound are:

¹ HNMR (400MHz, d-DMSO), δ12.23(s, 1H), 9.84(s, 1H), 8.1(d, J=4.8Hz, 1H), 7.33(t, J=16.8Hz, 2H), 6.72(d, J=8.4Hz, 2H), 6.18(s, 1H), 3.64(s, 6H), 2.21(s, 3H).

MS (ESI), m/z: 368 (M ⁺ +H ⁺ ).

Example 47 N ² , Preparation of N ⁴ -bis(4-fluoro-2-methylphenyl)thieno[3,2-d]pyrimidine-2,4-diamine (LD46011)

The chemical structural formula of N ² , N ⁴ -bis(4-fluoro-2-methylphenyl)thieno[3,2-d]pyrimidine-2,4-diamine is: Its synthesis steps are the same as in Example 1.

The characterization data of this compound are:

¹ HNMR (400MHz, d-DMSO), δ10.99(s, 1H), 9.82(s, 1H), 8.34(d, J=4.8Hz, 1H), 7.45(d, J=8.8Hz, 1H), 7.37(t, J=12.4Hz, 2H), 7.31(m, J=12Hz, 1H), 7.35(t, J=14.8Hz, 1H), 7.17(t, J=6.4Hz, 1H), 6.91(t , J=14.4Hz, 1H), 2.25(s, 3H), 2.18(s, 3H).

MS (ESI), m/z: 383 (M ⁺ +H ⁺ ).

Preparation of Example 48N ⁴ -phenyl-N ² -(5-fluoro-2-methylphenyl)thieno[3,2-d]pyrimidine-2,4-diamine (LD1149)

The chemical structural formula of N ⁴ -phenyl-N ² -(5-fluoro-2-methylphenyl)thieno[3,2-d]pyrimidine-2,4-diamine is: Its synthesis steps are the same as in Example 1.

The characterization data of this compound are:

¹ HNMR (400MHz, d-DMSO), δ8.38(s, 1H), 7.76(t, J=12Hz, 2H), 7.30(m, J=22.8Hz, 1H), 7.19(m, J=38.4Hz , 7H), 4.67(s, J=6Hz, 2H), 2.20(s, 3H).

MS (ESI), m/z: 365 (M ⁺ +H ⁺ ).

Example 49 2-(4-(3-chlorophenyl)piperazine-1-substituted)-N-(5-methyl-1H-pyrazole-3-substituted)thieno[3,2-d]pyrimidine- Preparation of 4-amine (LD4490)

2-(4-(3-chlorophenyl)piperazine-1-substituted)-N-(5-methyl-1H-pyrazole-3-substituted)thieno[3,2-d]pyrimidine-4- The chemical structural formula of an amine is: Its synthesis steps are the same as in Example 1.

The characterization data of this compound are:

¹ HNMR (400MHz, d-DMSO), δ12.12(s, 1H), 9.72(s, 1H), 7.93(d, J=5.2Hz, 1H), 7.22(t, J=16.4Hz, 1H), 7.09(d, J=5.2Hz, 1H), 7.01(d, J=1.6Hz, 1H), 6.96(m, J=10Hz, 1H), 6.35(s, 1H), 3.85(t, J=9.6Hz , 4H), 3.26(t, J=10Hz, 4H), 2.27(s, 3H).

MS (ESI), m/z: 427 (M ⁺ +H ⁺ ).

Example 50N ^2- (4-fluoro-3-(trifluoromethyl)benzyl)-N ⁴ -(5-methyl-1H-pyrazole-3-substituted)thieno[3,2-d]pyrimidine - Preparation of 2,4-diamine (LD4491)

N ² -(4-fluoro-3-(trifluoromethyl)benzyl)-N ⁴ -(5-methyl-1H-pyrazole-3-substituted)thieno[3,2-d]pyrimidine-2 , the chemical structural formula of 4-diamine is: Its synthesis steps are the same as in Example 1.

The characterization data of this compound are:

¹ HNMR (400MHz, d-DMSO), δ12.02(br, 1H), 9.61(br, 1H), 7.90(br, 1H), 7.73(t, J=24.8Hz, 2H), 7.43(t, J =19.6Hz, 1H), 7.22(br, 1H), 7.04(br, 1H), 6.34(br, 1H), 4.56(d, J=5.2Hz, 2H), 2.18(s, 3H).

MS (ESI), m/z: 423 (M ⁺ +H ⁺ ).

Example 51N ^4- (5-methyl-1H-pyrazole-3-substituted)-N ^2- (4-methylbenzyl)thieno[3,2-d]pyrimidine-2,4-diamine Preparation (LD4494)

The chemical structure of N ⁴ -(5-methyl-1H-pyrazole-3-substituted)-N ² -(4-methylbenzyl)thieno[3,2-d]pyrimidine-2,4-diamine for: Its synthesis steps are the same as in Example 1.

The characterization data of this compound are:

¹ HNMR (400MHz, d-DMSO), δ12.00(br, 1H), 9.74(br, 1H), 8.21(br, 1H), 7.89(br, 1H), 7.35(d, J=8Hz, 2H) , 7.23(d, J=6Hz, 2H), 7.04(br, 1H), 6.40(br, 1H), 4.49(d, J=6Hz, 2H), 2.25(s, 3H), 2.18(s, 3H) .

MS (ESI), m/z: 351 (M ⁺ +H ⁺ ).

Example 52 Preparation of N ² -(3-fluorobenzyl)-N ⁴ -(5-methyl-1H-pyrazole-3-substituted)thieno[3,2-d]pyrimidine-2,4-diamine (LD4497)

The chemical structure of N ² -(3-fluorobenzyl)-N ⁴ -(5-methyl-1H-pyrazole-3-substituted)thieno[3,2-d]pyrimidine-2,4-diamine is : Its synthesis steps are the same as in Example 1.

The characterization data of this compound are:

¹ HNMR (400MHz, d-DMSO), δ12.06(br, 1H), 9.58(br, 1H), 7.89(s, 1H), 7.35(m, J=39.2Hz, 1H), 7.07(m, J =64Hz, 4H), 6.61(br, 1H), 6.51(br, 1H), 3.54(d, J=6.4Hz, 2H), 2.20(s, 3H).

MS (ESI), m/z: 355 (M ⁺ +H ⁺ ).

Example 534-fluoro-N-(4-(4-(5-methyl-1H-pyrazole-3-substituted amine)thieno[3,2-d]pyrimidine-2-substituted amine)phenyl)benzene Formamide Preparation (LD4489)

4-fluoro-N-(4-(4-(5-methyl-1H-pyrazole-3-substituted amine)thieno[3,2-d]pyrimidine-2-substituted amine)phenyl)benzamide The synthetic route of is as follows:

Specifically include the following steps:

(1), N ² -(4-aminophenyl)-N ⁴ -(5-methyl-1H-pyrazole-3-substituted)thieno[3,2-d]pyrimidine-2,4-diamine , its chemical structure is: The synthesis steps are:

Add 2-chloro-N-(5-methyl-1H-pyrazole-3-substituted)thieno[3,2-d]pyrimidin-4-amine (0.3g, 1.13mmol) and benzene- 1,4-Diamine (0.367g, 3.39mmol), react at 145°C with the tube sealed until the reaction is complete. After the reaction was completed, the solvent was spin-dried, extracted with EtOAc and water, the organic phase was washed with saturated brine, dried over anhydrous Na ₂ SO ₄ , and separated by silica gel column chromatography to obtain a white solid which was N ² -(4-aminophenyl)- N ⁴ -(5-methyl-1H-pyrazole-3-substituted)thieno[3,2-d]pyrimidine-2,4-diamine (0.36 g, yield: 95%).

The characterization data of this compound are:

MS (ESI), m/z: 338 (M ⁺ +H ⁺ ).

(2), 4-fluoro-N-(4-(4-(5-methyl-1H-pyrazole-3-substituted amine)thieno[3,2-d]pyrimidine-2-substituted amine)phenyl ) Synthesis of benzamide, the chemical structural formula is: The synthesis steps are:

N ² -(4-aminophenyl)-N ⁴ -(5-methyl-1H-pyrazole-3-substituted)thieno[3,2-d]pyrimidine-2,4-diamine (150mg, 0.445mmol), 4-fluorobenzoic acid (68mg, 0.485mmol), HATU (183mg, 0.48mmol), DIPEA (1.85g, 0.25ml) were dissolved in DMF (1.0mL), stirred at room temperature until the reaction was complete. The reaction solution was extracted with DCM, washed with water, and the organic phases were combined and concentrated. The white solid obtained after column chromatography was 4-fluoro-N-(4-(4-(5-methyl-1H-pyrazole-3-substituted amine)thieno[3,2-d]pyrimidine-2-substituted amine )phenyl)benzamide (0.18g, 88% yield).

The characterization data of this compound are:

¹ HNMR (400MHz, d-DMSO), δ12.13(br, 1H), 10.13(br, 1H), 9.76(s, 1H), 9.02(s, 1H), 8.02(m, J=22.4Hz, 3H ), 7.80(d, J=8.8Hz, 2H), 7.63(s, 2H), 7.36(m, J=28.4Hz, 2H), 7.17(s, 1H), 6.56(br, 1H), 2.28(s , 3H).

MS (ESI), m/z: 460 (M ⁺ +H ⁺ ).

Example 54 Preparation of N-(5-methyl-1H-pyrazole-3-substituted)-2-(piperazine-1-substituted)thieno[3,2-d]pyrimidin-4-amine (LD4429)

The synthetic route of N-(5-methyl-1H-pyrazole-3-substituted)-2-(piperazine-1-substituted)thieno[3,2-d]pyrimidin-4-amine is as follows:

Specifically include the following steps:

(1), 4-(4-(5-methyl-1H-pyrazole-3-substituted amine)thieno[3,2-d]pyrimidine-2-substituted)piperazine-1-tert-butoxycarbonyl, Its chemical structural formula is: The synthesis steps are:

Add 2-chloro-N-(5-methyl-1H-pyrazole-3-substituted)thieno[3,2-d]pyrimidin-4-amine (0.3 g, 1.13 mmol) and piperazine to a reaction tube - 1-tert-butylformamide (0.633g, 3.39mmol), react at 140°C with the tube sealed until the reaction is complete. After the reaction was completed, the solvent was spin-dried, extracted with EtOAc and water, the organic phase was washed with saturated brine, dried over anhydrous Na ₂ SO ₄ , and separated by silica gel column chromatography to obtain a white solid which was 4-(4-(5-methyl- 1H-pyrazole-3-substituted amine)thieno[3,2-d]pyrimidine-2-substituted)piperazine-1-tert-butoxycarbonyl (0.32 g, yield: 90%).

The characterization data of this compound are:

MS(ESI), m/z:416(M ⁺ +H ⁺ ).

(2), Synthesis of N-(5-methyl-1H-pyrazole-3-substituted)-2-(piperazine-1-substituted)thieno[3,2-d]pyrimidin-4-amine, chemistry The structural formula is: The synthesis steps are:

4-(4-(5-methyl-1H-pyrazole-3-substituted amine)thieno[3,2-d]pyrimidine-2-substituted)piperazine-1-tert-butylformamide (0.32g, 0.77 mmol), TFA (0.7g, 0.5ml) was dissolved in dichloromethane (5ml). After stirring at room temperature for 2 hours, spin the solvent to dry, add saturated sodium bicarbonate solution to adjust the pH to alkaline, filter, and dry in vacuo to obtain a white solid that is N-(5-methyl-1H-pyrazole-3-substituted)-2 -(piperazine-1-substituted)thieno[3,2-d]pyrimidin-4-amine (0.22 g, yield: 92%).

The characterization data of this compound are:

¹ HNMR (400MHz, d-DMSO), δ12.10(brs, 1H), 9.64(brs, 1H), 7.89(t, J=16.4Hz, 1H), 7.05(m, J=20.4Hz, 1H), 6.28(s, 1H), 3.63(t, J=9.6Hz, 4H), 2.73(t, J=2.73Hz, 4H), 2.23(s, 3H).

MS (ESI), m/z: 316 (M ⁺ +H ⁺ ).

Example 55 2-(4-fluorophenyl)-1-(4-(4-(5-methyl-1H-pyrazole-3-substituted amine)thieno[3,2-d]pyrimidine-2-substituted ) Preparation of piperidine-1-substituted) ethyl ketone (LD4456)

2-(4-fluorophenyl)-1-(4-(4-(5-methyl-1H-pyrazole-3-substituted amine)thieno[3,2-d]pyrimidine-2-substituted)piper The synthetic route of pyridine-1-substituted) ethyl ketone is as follows:

Specifically include the following steps:

(1), Synthesis of N-(5-methyl-1H-pyrazole-3-substituted)-2-(piperazine-1-substituted)thieno[3,2-d]pyrimidin-4-amine, the steps are the same Example 54;

(2), N-(5-methyl-1H-pyrazole-3-substituted)-2-(piperazine-1-substituted)thieno[3,2-d]pyrimidin-4-amine (100mg, 0.317mmol), 2-(4-fluorophenyl) acetic acid (54mg, 0.35mmol), HATU (145mg, 0.38mmol), DIPEA (1.48g, 0.2ml) was dissolved in the mixed solvent of DMF/DCM=1 (2.0 mL), stirred at room temperature until the reaction was complete. The reaction solution was extracted with DCM, washed with water, and the organic phases were combined and concentrated. The white solid after column chromatography was 2-(4-fluorophenyl)-1-(4-(4-(5-methyl-1H-pyrazole-3-substituted amine)thieno[3,2-d] Pyrimidine-2-substituted)piperidine-1-substituted)ethanone (0.122g, 85% yield).

The characterization data of this compound are:

¹ HNMR (400MHz, d-DMSO), δ12.11(s, 1H), 9.71(s, 1H), 7.93(d, J=5.2Hz, 1H), 7.36(d, J=8.4Hz, 2H), 7.27(d, J=8.4Hz, 2H), 7.08(d, J=5.2Hz, 1H), 6.3(s, 1H), 3.78(s, 2H), 3.69(s, 4H), 3.58(t, J =12.8Hz, 4H), 2.25(s, 3H).

MS (ESI), m/z: 452 (M ⁺ +H ⁺ ).

Example 56 (3-chlorophenyl) (4-(4-(5-methyl-1H-pyrazole-3-substituted amine)thieno[3,2-d]pyrimidine-2-substituted)piperazine- 1-substituted) ketone preparation (LD4449)

(3-chlorophenyl)(4-(4-(5-methyl-1H-pyrazole-3-substituted amine)thieno[3,2-d]pyrimidine-2-substituted)piperazine-1-substituted ) The chemical structural formula of methyl ketone is: Its synthesis steps are the same as in Example 55.

The characterization data of this compound are:

¹ HNMR (400MHz, d-DMSO), δ12.10(s, 1H), 9.72(s, 1H), 7.94(d, J=5.2Hz, 1H), 7.51(m, J=30.8Hz, 3H), 7.41(d, J=7.6Hz, 1H), 7.09(d, J=5.2Hz, 1H), 6.29(s, 1H), 3.78(m, J=38.4Hz, 8H), 2.23(s, 3H).

MS (ESI), m/z: 454 (M ⁺ +H ⁺ ).

Example 57 (4-chlorophenyl)-(4-(4-(5-methyl-1H-pyrazole-3-substituted amine)thieno[3,2-d]pyrimidine-2-substituted)piperidine -1-substituted) ketone preparation (LD4455)

(4-chlorophenyl)-(4-(4-(5-methyl-1H-pyrazole-3-substituted amine)thieno[3,2-d]pyrimidine-2-substituted)piperidine-1- Substitution) The chemical structural formula of methyl ketone is: Its synthesis steps are the same as in Example 55.

The characterization data of this compound are:

¹ HNMR (400MHz, d-DMSO), δ12.09(s, 1H), 9.72(s, 1H), 7.94(d, J=1.2Hz, 1H), 7.51(m, J=28.8Hz, 4H), 7.09 (d, J=5.2Hz, 1H), 6.3 (s, 1H), 3.81 (s, 2H), 3.74 (s, 4H), 3.43 (s, 2H), 2.23 (s, 3H).

MS (ESI), m/z: 454 (M ⁺ +H ⁺ ).

Example 58 (5-chloro-2-fluorophenyl)-(4-(4-(5-methyl-1H-pyrazole-3-substituted amine)thieno[3,2-d]pyrimidine-2- Preparation of substituted) piperidine-1-substituted) ketones (LD4463)

(5-chloro-2-fluorophenyl)-(4-(4-(5-methyl-1H-pyrazole-3-substituted amine)thieno[3,2-d]pyrimidine-2-substituted)piper The chemical structural formula of pyridine-1-substituted)methanone is: Its synthesis steps are the same as in Example 55.

The characterization data of this compound are:

¹ HNMR (400MHz, d-DMSO), δ12.10(s, 1H), 9.74(s, 1H), 7.94(d, J=5.2Hz, 1H), 7.58(m, J=15.6Hz, 2H), 7.39(m, J=18Hz, 1H), 7.09(d, J=5.2Hz, 1H), 6.29(s, 1H), 3.82(d, J=5.6Hz, 2H), 3.73(s, 4H), 3.31 (s, 2H), 2.23 (s, 3H).

MS (ESI), m/z: 473 (M ⁺ +H ⁺ ).

Example 59 2-(2-chlorophenyl)-1-(4-(4-(5-methyl-1H-pyrazole-3-substituted amine)thieno[3,2-d]pyrimidine-2-substituted ) Preparation of piperidine-1-substituted) ethyl ketone (LD4469)

2-(2-chlorophenyl)-1-(4-(4-(5-methyl-1H-pyrazole-3-substituted amine)thieno[3,2-d]pyrimidine-2-substituted)piper The chemical structural formula of pyridine-1-substituted) ethyl ketone is: Its synthesis steps are the same as in Example 55.

The characterization data of this compound are:

¹ HNMR (400MHz, d-DMSO), δ12.11(s, 1H), 9.74(s, 1H), 7.94(d, J=5.2Hz, 1H), 7.42(m, J=8.8Hz, 1H), 7.30(m, J=26.8Hz, 3H), 7.1(d, J=5.2Hz, 1H), 6.31(s, 1H), 3.88(s, 2H), 3.76(d, J=21.2Hz, 4H), 3.62(d, J=29.2Hz, 4H), 2.25(s, 3H).

MS (ESI), m/z: 469 (M ⁺ +H ⁺ ).

Example 60 (4-(4-(5-methyl-1H-pyrazole-3-substituted amine)thieno[3,2-d]pyrimidine-2-substituted)piperidine-1-substituted)(1- Preparation of phenylcyclopropanyl) ketone (LD4474)

(4-(4-(5-methyl-1H-pyrazole-3-substituted amine)thieno[3,2-d]pyrimidine-2-substituted)piperidine-1-substituted)(1-phenyl ring The chemical structural formula of propanyl) ketone is: Its synthesis steps are the same as in Example 55.

The characterization data of this compound are:

¹ HNMR (400MHz, d-DMSO), δ12.12(s, 1H), 9.75(s, 1H), 8.16(s, 1H), 7.93(d, J=5.2Hz, 1H), 7.32(t, J =15.2Hz, 2H), 7.20(t, J=14.8Hz, 2H), 7.06(d, J=14.8Hz, 1H), 6.22(s, 1H), 3.63(m, J=36.4Hz, 4H), 3.32(s, 4H), 3.13(m, J=26.4Hz, 2H), 2.23(s, 3H), 1.25(m, J=36.4Hz, 2H).

MS (ESI), m/z: 460 (M ⁺ +H ⁺ ).

Example 61 (4-bromophenyl)-(4-(4-(5-methyl-1H-pyrazole-3-substituted amine)thieno[3,2-d]pyrimidine-2-substituted)piperidine -1-substituted) ketone preparation (LD4454)

(4-bromophenyl)-(4-(4-(5-methyl-1H-pyrazole-3-substituted amine)thieno[3,2-d]pyrimidine-2-substituted)piperidine-1- Substitution) The chemical structural formula of methyl ketone is: Its synthesis steps are the same as in Example 55.

The characterization data of this compound are:

¹ HNMR (400MHz, d-DMSO), δ12.09(s, 1H), 9.73(s, 1H), 7.94(d, J=5.2Hz, 1H), 7.67(d, J=8.4Hz, 1H), 7.41(d, J=8Hz, 2H), 7.09(d, J=5.6Hz, 1H), 6.29(s, 1H), 3.81(s, 2H), 3.80(s, 4H), 3.42(s, 2H) , 2.23(s, 3H).

MS (ESI), m/z: 499 (M ⁺ +H ⁺ ).

Example 62 (4-(4-(5-methyl-1H-pyrazole-3-substituted amine)thieno[3,2-d]pyrimidine-2-substituted)piperazine-1-substituted)(pyridine- Preparation of 2-substituted)methanones (LD4446)

(4-(4-(5-methyl-1H-pyrazole-3-substituted amine)thieno[3,2-d]pyrimidine-2-substituted)piperazine-1-substituted)(pyridine-2-substituted ) The chemical structural formula of methyl ketone is: Its synthesis steps are the same as in Example 55.

The characterization data of this compound are:

¹ HNMR (400MHz, d-DMSO), δ12.10(s, 1H), 9.72(s, 1H), 8.62(d, J=4.8Hz, 1H), 7.72(m, J=56.8Hz, 2H), 7.61(d, J=8Hz, 1H), 7.50(m, J=12.4Hz, 1H), 7.09(d, J=5.6Hz, 1H), 6.30(s, 1H), 3.84(s, 2H), 3.74 (s, 4H), 3.64 (s, 4H), 2.23 (s, 3H).

MS (ESI), m/z: 421 (M ⁺ +H ⁺ ).

Example 631-(4-(4-(5-methyl-1H-pyrazole-3-substituted amine)thieno[3,2-d]pyrimidine-2-substituted)piperazine-1-substituted)-3 - Preparation of phenyl-1-propanone (LD4448)

1-(4-(4-(5-methyl-1H-pyrazole-3-substituted amine)thieno[3,2-d]pyrimidine-2-substituted)piperazine-1-substituted)-3-benzene The chemical structural formula of base-1-propanone is: Its synthesis steps are the same as in Example 55.

The characterization data of this compound are:

¹ HNMR (400MHz, d-DMSO), δ12.13(s, 1H), 9.75(s, 1H), 7.94(d, J=5.2Hz, 1H), 7.27(m, J=19.6Hz, 4H), 7.17(m, J=17.2Hz, 1H), 7.09(d, J=5.2Hz, 1H), 6.29(s, 1H), 3.67(s, 4H), 3.52(d, J=16.4Hz, 4H), 2.84(t, J=15.6Hz, 2H), 22.68(t, J=15.6Hz, 2H), 2.25(s, 3H).

MS (ESI), m/z: 448 (M ⁺ +H ⁺ ).

Example 64 (4-(4-(5-methyl-1H-pyrazole-3-substituted amine)thieno[3,2-d]pyrimidine-2-substituted)piperazine-1-substituted)(pyridine- Preparation of 4-substituted) ketones (LD4450)

(4-(4-(5-methyl-1H-pyrazole-3-substituted amine)thieno[3,2-d]pyrimidine-2-substituted)piperazine-1-substituted)(pyridine-4-substituted ) The chemical structural formula of methyl ketone is: Its synthesis steps are the same as in Example 55.

The characterization data of this compound are:

¹ HNMR (400MHz, d-DMSO), δ12.10(s, 1H), 9.72(s, 1H), 8.68(m, J=5.6Hz, 2H), 7.96(d, J=5.6Hz, 2H), 7.43(m, J=6Hz, 2H), 7.09(d, J=5.2Hz, 1H), 6.28(s, 1H), 3.84(s, 1H), 3.73(s, 4H), 3.08(m, J= 22Hz, 4H), 2.22(s, 1H).

MS (ESI), m/z: 421 (M ⁺ +H ⁺ ).

Example 65 (4-(4-(5-methyl-1H-pyrazole-3-substituted amine)thieno[3,2-d]pyrimidine-2-substituted)piperazine-1-substituted)(pyrazine -2-substituted)methanone preparation (LD4447)

(4-(4-(5-methyl-1H-pyrazole-3-substituted amine)thieno[3,2-d]pyrimidine-2-substituted)piperazine-1-substituted)(pyrazine-2- The chemical structural formula of substituted) ketone is: Its synthesis steps are the same as in Example 55.

The characterization data of this compound are:

¹ HNMR (400MHz, d-DMSO), δ12.11(s, 1H), 9.73(s, 1H), 8.88(d, J=1.2Hz, 1H), 8.76(d, J=2.4Hz, 1H), 8.70(d, J=3.6Hz, 2H), 7.94(d, J=5.6Hz, 1H), 7.09(d, J=5.2Hz, 1H), 6.30(s, 1H), 3.86(d, J=5.2 Hz, 2H), 3.76(s, 4H), 3.53(s, 2H), 2.23(s, 3H).

MS (ESI), m/z: 422 (M ⁺ +H ⁺ ).

Example 661-(4-(4-(5-methyl-1H-pyrazole-3-substituted amine)thieno[3,2-d]pyrimidine-2-substituted)piperidine-1-substituted)propane- Preparation of 1-keto (LD4459)

1-(4-(4-(5-methyl-1H-pyrazole-3-substituted amine)thieno[3,2-d]pyrimidine-2-substituted)piperidine-1-substituted)propane-1- The chemical structural formula of a ketone is: Its synthesis steps are the same as in Example 55.

The characterization data of this compound are:

¹ HNMR (400MHz, d-DMSO), δ12.12(s, 1H), 9.73(s, 1H), 9.62(s, 1H), 7.94(d, J=5.2Hz, 1H), 7.09(d, J =5.2Hz, 1H), 6.31(s, 1H), 3.72(d, J=23.6Hz, 4H), 3.53(s, 4H), 2.37(m, J=22Hz, 2H), 2.25(s, 3H) , 0.93 (m, J=49.2Hz, 3H).

MS (ESI), m/z: 372 (M ⁺ +H ⁺ ).

Example 67N ^4- (5-methyl-1H-pyrazole-3-substituted)-N ² -(3-methylbenzyl)thieno[3,2-d]pyrimidine-2,4-diamine salt Preparation of acid salt (LD4484 hydrochloride)

N ⁴ -(5-methyl-1H-pyrazole-3-substituted)-N ² -(3-methylbenzyl)thieno[3,2-d]pyrimidine-2,4-diamine hydrochloride The chemical structure formula is: The synthesis steps are:

N ⁴ -(5-methyl-1H-pyrazole-3-substituted)-N ² -(3-methylbenzyl)thieno[3,2-d]pyrimidine-2,4-diamine (0.351 g, 1mmol) into a 100mL three-neck flask, add 20mL of absolute ethanol, add dropwise 6mol/L hydrochloric acid aqueous solution under stirring, wait until all the raw materials are dissolved, spin dry to obtain a white solid, namely N ⁴ -(5-methyl-1H- Pyrazole-3-substituted)-N ² -(3-methylbenzyl)thieno[3,2-d]pyrimidine-2,4-diamine hydrochloride (0.405 g, yield 90%).

The characterization data of this compound are:

¹ HNMR (400MHz, d-DMSO), δ8.00(br, 4H), 7.43(t, 1H), 7.20(d, 1H), 6.96～7.10(m, 6H), 5.00(d, 1H), 4.59 (s,2H), 2.34(s,3H), 1.71(d,3H).

MS (ESI), m/z: 351 (M ⁺ +H ⁺ ).

Example 68 (4-(4-(5-methyl-1H-pyrazole-3-substituted amine)thieno[3,2-d]pyrimidine-2-substituted)piperazine-1-substituted)(pyridine- Preparation of 4-substituted) ketone hydrochloride (LD4450 hydrochloride)

(4-(4-(5-methyl-1H-pyrazole-3-substituted amine)thieno[3,2-d]pyrimidine-2-substituted)piperazine-1-substituted)(pyridine-4-substituted ) The chemical structural formula of ketone hydrochloride is: Its synthesis steps are the same as in Example 67.

The characterization data of this compound are:

¹ HNMR (400MHz, d-DMSO), δ8.89(d, 2H), 8.0(br, 3H), 7.92(d, 2H), 7.20(d, 1H), 7.0(br, 3H), 6.96(d , 1H), 5.0 (s, 1H), 4.07 (s, 8H), 1.71 (d, 3H).

MS (ESI), m/z: 421 (M ⁺ +H ⁺ ).

Example 69 2-(4-fluorophenyl)-1-(4-(4-(5-methyl-1H-pyrazole-3-substituted amine)thieno[3,2-d]pyrimidine-2-substituted ) piperidine-1-substituted) ethyl ketone mesylate preparation (LD4456 mesylate)

2-(4-fluorophenyl)-1-(4-(4-(5-methyl-1H-pyrazole-3-substituted amine)thieno[3,2-d]pyrimidine-2-substituted)piper The chemical structural formula of pyridine-1-substituted) ethyl ketone mesylate is: The synthesis steps are:

2-(4-fluorophenyl)-1-(4-(4-(5-methyl-1H-pyrazole-3-substituted amine)thieno[3,2-d]pyrimidine-2-substituted) Piperidine-1-substituted) ethyl ketone (0.451g, 1mmol) was placed in a 100mL single-necked bottle, 30mL of absolute ethanol was added, 384mg of methanesulfonic acid (4mmol) was added dropwise under stirring, heated to boiling, the system became clear, and refluxed for 4h , cooled to room temperature, a white solid precipitated out, filtered, the filter residue was washed three times with ethanol, and dried in vacuo to obtain 0.750 g (90%) of a pale yellow solid.

The characterization data of this compound are:

¹ HNMR (400MHz, d-DMSO), δ8.00(br, 3H), 7.34(m, 2H), 7.20(d, 1H), 7.12(m, 2H), 7.0(br, 3H), 6.96(ds , 1H), 5.00 (d, 1H), 4.07 (s, 8H), 3.66 (s, 2H), 2.84 (s, 12H), 1.71 (d, 3H).

MS (ESI), m/z: 452 (M ⁺ +H ⁺ ).

Example 70N ^2- (3-bromobenzyl)-N ⁴ -(5-methyl-1H-pyrazole-3-substituted)thieno[3,2-d]pyrimidine-2,4-diaminomethanesulfonate Preparation of acid salt (LD4481 mesylate)

N ² -(3-bromobenzyl)-N ⁴ -(5-methyl-1H-pyrazole-3-substituted)thieno[3,2-d]pyrimidine-2,4-diamine methanesulfonate The chemical structure formula is: Its synthesis steps are the same as in Example 69.

The characterization data of this compound are:

H ¹ NMR (400MHz, d-DMSO), δ8.00(br, 4H), 7.22～7.41(m, 3H), 7.17～7.20(m, 2H), 7.00(br, 2H), 6.96(d, 1H ), 5.00 (d, 1H), 4.59 (s, 2H), 2.84 (s, 9H), 1.71 (d, 3H).

MS (ESI), m/z: 416 (M ⁺ +H ⁺ ).

Example 71 In vitro kinase activity detection

Kinase activity was detected by fluorescence resonance energy transfer (FRET) Z-LYTE method. The drug is serially diluted (3-fold dilution starting from 10 μM), and the kinase (about 0.1-0.01U/mL for AuroraA or about 0.1-0.01U/mL for AuroraB) and coumarin (fluorescence donor) and fluorophores ( Fluorescence acceptor) substrate (20μM) into the reaction system of 10ul, add the final concentration of 10uMATP and react at room temperature for 2 hours; then add 5ul of Development solution to the reaction system, react at room temperature for 1 hour, add 5ul of Then, detect it on a microplate reader. Under the excitation wavelength of 400nm, the ratios obtained by detecting the emission wavelengths at 460nm and 535nm of the emission light respectively were subjected to fitting analysis by GraphpadPrism5.0 to obtain the IC ₅₀ of the screened drug. The IC ₅₀ values of the half maximal inhibitory concentration of each compound against AuroraA or AuroraB kinase are described in Table 1. The compounds used were selected from the compounds prepared for Examples 1-66, respectively.

The compound of table 1. embodiment 1-66 is to AuroraA, B kinase activity IC ₅₀ (nM)

(ND: Not Detected, NR: Not Reported)

Combining the chemical structures of the compounds prepared in Examples 1-66 and the in vitro kinase activity data in Table 1, it can be analyzed that thieno-2,4-substituted pyrimidine compounds have a significant structure-activity relationship with AuroraA kinase and AuroraB kinase. Its characteristics are as follows: 1) When X ₂ is NH or NCH ₃ , when R ₂ is 5-methyl-1H-pyrazolyl, its activity is better than other molecules; 2) When X ₂ is NH or NCH ₃ , R ₂ is 5-methyl-1H-pyrazolyl, and when X ₁ is NH or NCH ₃ , when R ₁ is substituted phenyl or benzyl, its activity is better than other molecules; 3) phenyl or benzyl The size of the substituent affects the activity of the compound. The general rule is that the activity of fewer substituents is better than that of multiple substituents, and the activity of small substituents is better than that of large substituents; 4) When X ₁ is NH or NCH ₃ , R ₁ is 5-methyl 5) The longer the alkyl chain in the substituent, the worse the inhibitory activity of Aurora kinase; 6) The sequence of the four structural formula compounds inhibiting Aurora kinase activity Generally, the formula (I) > (III) > (II) >(IV); 7) When X ₃ and X ₄ are N, the compound can inhibit Aurora kinase activity better than the compound X ₃ and X ₄ are O and CH .

The structure of the compound with better activity is as follows:

The chemical structure of LD4484 is: The chemical structure of LD4481 is:

The chemical structure of LD4478 is: The chemical structure of LD4456 is:

Example 72 In Vitro Cell Viability Detection

The cytostatic activity of thieno 2,4 substituted pyrimidines was assessed using the method described in the CCK-8 kit. Cells (3000-10000/well) were seeded on 96-well cell culture plates for 24 hours, 100 μL of compound solutions of different concentrations were added to each culture well, incubated for 72 hours, 10 μL CCK-8 solution was added to each culture well, Incubate for another 2-3 hours, and measure the absorbance at 450 nm and 650 nm with a microplate reader. The raw data were processed in an Excel spreadsheet to obtain the cell viability of each well treated. _IC50 values were then calculated using non-linear regression models on GraphPad Prism software using the survival data. It was found that some thieno 2,4 substituted pyrimidine compounds could significantly inhibit Hela (human cervical tumor cells), HT1080 (human fibrosarcoma cells), MCF-7 (breast cancer cells), A431 (human skin basal cell carcinoma cells) ), H1975 (non-small cell lung cancer cells), BT474 (human breast ductal tumor cells), U937 (human macrophage leukemia tumor cells), MOLT-4 (human T lymphocyte line tumor cells), half of the The inhibitory concentration IC ₅₀ is positively correlated with the drug concentration. The compounds used were those prepared in Examples 1-66, and the results are shown in Table 2.

Table 2. IC ₅₀ (uM) of the compounds of Examples 1-66 on different tumor cells

Combining the chemical structures of the compounds prepared in Examples 1-66 and the in vitro cell activity data given in Table 2, it can be analyzed that the structure-activity relationship of thieno-2,4-substituted pyrimidine compounds on in vitro cell activity is basically consistent with that in Example 67 . It can be seen from Table 2 that some compounds have strong inhibitory activity on Hela (cervical tumor cells), U937 (human macrophage leukemia cells), and MoLT-4 (human T lymphocyte line cancer cells).

It can be deduced from Table 2 that the thieno-2,4-substituted pyrimidine compounds with the structure of formula I can treat human or other mammalian breast cancer, respiratory tract cancer, brain cancer, tumors of male and female reproductive organs and digestive tract Tumors, urethral tumors, liver cancer, skin cancer, head and neck cancer, lymphoma, sarcoma, and leukemia, including but not limited to breast cancer such as invasive ductal carcinoma, mucinous carcinoma, lobular invasive carcinoma, tubular carcinoma, cystic adenoid carcinoma, papillary Carcinoma, small cell bronchial carcinoma (oat cell carcinoma), non-small cell bronchial carcinoma such as plateepithelial carcinoma, adenocarcinoma, large cell bronchial carcinoma, pleuropulmonary blastoma, brainstem and subocular glioma, cerebellar and cerebral astrocytes Ependymal tumors, ependymal tumors and neuroectodermal and pineal tumors, prostate and testicular cancers such as seminoma, nonseminoma, endometrial cancer, cervical cancer, ovarian cancer such as mucinous, intrauterine Membranous, or serum cancer, vaginal cancer, vulvar cancer and intrauterine tumor, anal cancer, colon cancer, colorectal cancer, esophagus cancer, stomach cancer, pancreatic cancer rectal cancer, small intestine cancer or salivary gland cancer, bladder cancer, penile cancer , kidney, renal pelvis, ureter or urethra, intraocular melanoma and retinoblastoma, hepatoma (stem cell cancer with or without fibrolamellar changes), cholangiocarcinoma (intrahepatic bile duct cancer), and mixed hepatic Cellular cholangiocarcinoma, squamous cell carcinoma, Kaposi's sarcoma, malignant melanoma, Merkel's cell skin and non-melanoma cell carcinomas, larynx, hypopharynx, nasopharynx, oropharynx, and lip and oral cavity cancers , AIDS-related lymphoma, non-Hodgkin lymphoma, cutaneous T-cell lymphoma, Hodgson disease and central nervous system lymphoma, soft tissue sarcoma, osteosarcoma, malignant fibrous histiocytoma, lymphosarcoma and rhabdomyosarcoma, acute Myeloid leukemia, acute forest cell leukemia, chronic lymphocytic leukemia, chronic myelogenous leukemia, and hairy cell leukemia.

The above-mentioned embodiments only express several implementation modes of the present invention, and the descriptions thereof are relatively specific and detailed, but should not be construed as limiting the patent scope of the present invention. It should be noted that those skilled in the art can make several modifications and improvements without departing from the concept of the present invention, and these all belong to the protection scope of the present invention. Therefore, the protection scope of the patent for the present invention should be based on the appended claims.

Claims (6)

1. general formula is thieno-2,4 substituted pyrimidines compounds or its pharmacy acceptable salt of (I):

Wherein, X ₁, X ₂be O, S, S (O), S (O) simultaneously or independently ₂or NR ₅, wherein R ₅for H or C _1-6alkyl;

R ₁, R ₂simultaneously or be independently selected from following groups:

Wherein, f is the integer between 0-3, and dotted line represents and X ₁, X ₂the key connected;

R ₆₁, R ₆₂, R ₆₃be hydrogen, C simultaneously or independently _1-5alkyl, halogen, nitro, cyano group, amino or hydroxyl.

2. thieno-2,4 substituted pyrimidines compounds according to claim 1 or its pharmacy acceptable salt, is characterized in that, described X ₁with described X ₂be NH or NCH simultaneously or independently ₃.

3. thieno-2,4 substituted pyrimidines compounds according to claim 1 or its pharmacy acceptable salt, is characterized in that, described R ₁, R ₂simultaneously or be independently

4. thieno-2,4 substituted pyrimidines compounds according to claim 1 or its pharmacy acceptable salt, is characterized in that, be selected from one of following compound:

N ²-(the fluoro-3-of 4-(trifluoromethyl) benzyl)-N ⁴-(5-methyl isophthalic acid H-pyrazoles-3-replaces) thieno-[3,2-d] pyrimidine-2,4-diamines;

N ⁴-(5-methyl isophthalic acid H-pyrazoles-3-replaces)-N ²-(4-methyl-benzyl) thieno-[3,2-d] pyrimidine-2,4-diamines;

N ⁴-(5-methyl isophthalic acid H-pyrazoles-3-replaces)-N ²-(3-methyl-benzyl) thieno-[3,2-d] pyrimidine-2,4-diamines;

N ²-(3-luorobenzyl)-N ⁴-(5-methyl isophthalic acid H-pyrazoles-3-replaces) thieno-[3,2-d] pyrimidine-2,4-diamines;

N ²-(3-bromobenzyl)-N ⁴-(5-methyl isophthalic acid H-pyrazoles-3-replaces) thieno-[3,2-d] pyrimidine-2,4-diamines;

N ²-(the bromo-2-luorobenzyl of 4-)-N ⁴-(5-methyl isophthalic acid H-pyrazoles-3-replaces) thieno-[3,2-d] pyrimidine-2,4-diamines;

N ²-((2-chloropyridine-4-replaces) methyl)-N ⁴-(5-methyl isophthalic acid H-pyrazoles-3-replaces) thieno-[3,2-d] pyrimidine-2,4-diamines;

N ²-(2-luorobenzyl)-N ⁴-(5-methyl isophthalic acid H-pyrazoles-3-replaces) thieno-[3,2-d] pyrimidine-2,4-diamines;

N ²-(4-chlorobenzyl)-N ⁴-(5-methyl isophthalic acid H-pyrazoles-3-replaces) thieno-[3,2-d] pyrimidine-2,4-diamines;

N ²-(the chloro-6-luorobenzyl of 2-)-N ⁴-(5-methyl isophthalic acid H-pyrazoles-3-replaces) thieno-[3,2-d] pyrimidine-2,4-diamines;

N ²-(2,6-dichloro benzyl)-N ⁴-(5-methyl isophthalic acid H-pyrazoles-3-replaces) thieno-[3,2-d] pyrimidine-2,4-diamines;

N ⁴-(5-methyl isophthalic acid H-pyrazoles-3-replaces)-N ²-(4-(trifluoromethyl) benzyl) thieno-[3,2-d] pyrimidine-2,4-diamines;

N ²-(4-chlorobenzene ethyl)-N ⁴-(5-methyl isophthalic acid H-pyrazoles-3-replaces) thieno-[3,2-d] pyrimidine-2,4-diamines;

N ²-(2,4 difluorobenzene base)-N ⁴-(5-methyl isophthalic acid H-pyrazoles-3-replaces) thieno-[3,2-d] pyrimidine-2,4-diamines;

N ²-(3-bromophenyl)-N ⁴-(5-methyl isophthalic acid H-pyrazoles-3-replaces) thieno-[3,2-d] pyrimidine-2,4-diamines;

N ²-(xenyl-4-replaces)-N ⁴-(5-methyl isophthalic acid H-pyrazoles-3-replaces) thieno-[3,2-d] pyrimidine-2,4-diamines;

N ²-(the chloro-3-of 4-(trifluoromethyl) phenyl)-N ⁴-(5-methyl isophthalic acid H-pyrazoles-3-replaces) thieno-[3,2-d] pyrimidine-2,4-diamines;

N ²-(4-bromine-3-fluorophenyl)-N ⁴-(5-methyl isophthalic acid H-pyrazoles-3-replaces) thieno-[3,2-d] pyrimidine-2,4-diamines;

N ²-(the fluoro-5-of 3-(trifluoromethyl) phenyl)-N ⁴-(5-methyl isophthalic acid H-pyrazoles-3-replaces) thieno-[3,2-d] pyrimidine-2,4-diamines;

N ²-(2,6-diethyl phenyl)-N ⁴-(5-methyl isophthalic acid H-pyrazoles-3-replaces) thieno-[3,2-d] pyrimidine-2,4-diamines;

2-(2,6-Dimethoxyphenyl)-N-(5-methyl isophthalic acid H-pyrazoles-3-replaces) thieno-[3,2-d] pyrimidine-4-amine;

N ⁴-(5-methyl isophthalic acid H-pyrazoles-3-replaces)-N ²-(3-methyl-benzyl) thieno-[3,2-d] pyrimidine-2,4-diamine hydrochloride;

N ²-(3-bromobenzyl)-N ⁴-(5-methyl isophthalic acid H-pyrazoles-3-replaces) thieno-[3,2-d] pyrimidine-2,4-diamines mesylate.

5. be used for the treatment of a pharmaceutical composition for tumour, it is characterized in that, be made up of arbitrary described thieno-2, the 4 substituted pyrimidines compounds of claim 1-4 or its pharmacy acceptable salt and pharmaceutically acceptable carrier.

6. according to arbitrary described thieno-2, the 4 substituted pyrimidines compounds of claim 1-4 or its pharmacy acceptable salt preparing anti-tumor drugs targeting, prevent and/or treat cancer, infection, inflammation and autoimmune disease pharmaceutical composition in application.