A diaryl urea PI3K / mTOR / HDAC multi-target inhibitor and its pharmaceutical composition and application

Abstract

The present invention belongs to the field of medical technology, and specifically relates to a diaryl urea PI3K / mTOR / HDAC multi-target inhibitor and its pharmaceutical composition and application. The diaryl urea PI3K / mTOR / HDAC multi-target inhibitor of the present invention belongs to a single-molecule multi-target inhibitor, has multiple mechanisms of action, and has the characteristics of good efficacy and low toxicity. This type of compound has the characteristics of effectively inhibiting multiple PI3K subtype kinases, mTOR kinases and HDAC proteins. The structure of the diaryl urea PI3K / mTOR / HDAC multi-target inhibitor is as follows:

Description

Technical Field

[0001] The present invention belongs to the field of medical technology, and specifically relates to a diaryl urea PI3K / mTOR / HDAC multi-target inhibitor and a pharmaceutical composition and application thereof. Background Art

[0002] The occurrence and development of tumors are closely related to cell signal transduction pathways, and their abnormal activation usually leads to the formation of tumors. Drugs designed based on key regulatory proteins in cell signal transduction pathways have good in vivo safety due to their high targeting and selectivity. Therefore, anti-tumor drugs targeting key kinases in signaling pathways have become an important direction of drug research in recent years.

[0003] The phosphatidylinositol-3 kinase (PI3K)-target of rapamycin (mTOR) signaling pathway is widely present in mammalian cells and regulates a variety of cellular life activities such as cell growth, proliferation, differentiation and survival. It is also one of the most frequently activated pathways in human cancer, affecting nearly 50% of malignant tumors. Among them, PI3K is a member of the serine / threonine lipid kinase family and can be activated by G protein-coupled receptors and tyrosine kinase receptors, as well as by Ras proteins. According to its sequence homology, structural characteristics and substrate molecular specificity, PI3K is divided into type I, type II and type III PI3K, among which type I PI3K is the most studied. According to the type of cell surface receptor and catalytic domain, type I PI3K is further subdivided into IA (PI3Kα, β and δ) and IB (PI3Kγ). Type I PI3K can regulate cell metabolism and growth by phosphorylating phosphatidylinositol at position 3 to produce phosphatidylinositol triphosphate (PIP3). The lipid's second messenger PIP3 can pair P13K with downstream effectors (especially Akt), leading to membrane recruitment and phosphorylation, as well as cell proliferation and activation. mTOR is also a serine / threonine protein kinase and a member of the PI3K-related kinase family (PIKK). It exists in the body in the form of two complexes, namely mTORC1 and mTORC2. Both complexes can regulate cell growth, but play different roles in cells. mTORC1 is located downstream of Akt kinase and can directly phosphorylate ribosomal protein S6 kinase 1 and eukaryotic translation initiation factor 4E binding protein 1, thereby regulating gene transcription, mRNA splicing and protein synthesis. mTORC2 upregulates the PI3K / AKt / mTOR (PAM) signaling pathway by phosphorylating Akt's Ser473 site, thereby promoting the growth of tumor cells. As a key kinase in this pathway, the overactivation of mTOR is closely related to the poor prognosis of cancer.

[0004] The PI3K / mTOR signaling pathway is involved in regulating various processes of cell life activities. It not only promotes the occurrence and development of tumors, invasion and metastasis, and tumor angiogenesis, but also promotes glycogen synthesis and glycolysis, and increases glucose uptake. Mutation or overexpression of tyrosine kinases and functional loss of the tumor suppressor PTEN will lead to abnormal activation of the PAM signaling pathway and induce tumors. Therefore, organic small molecule kinase inhibitors targeting kinases such as PI3K and mTOR in this pathway have become a hot spot in the development of molecular targeted anti-tumor drugs.

[0005] The research on PI3K inhibitors has a history of 20 years. Nearly 30 candidate drugs have entered clinical trials, half of which have entered Phase II clinical trials. Given the close relationship between PI3K inhibitors and malignant tumors and kinase systems, their inhibitors have become potential anti-cancer drugs. The following six PI3K inhibitors have been approved for marketing by the U.S. Food and Drug Administration (FDA).

[0006]

[0007] The first generation of mTOR inhibitors can bind to the FRB region by forming a complex with FKBP12, and regulate the function of the target enzyme through an allosteric mechanism. Among them, rapamycin and its structural analogs Everolimus and Temsirolimus have been approved for the treatment of breast cancer and advanced renal cancer, which fully confirms the application prospects of mTOR targeted therapy in anti-tumor. However, the above mTOR inhibitors can only inhibit mTORC1, and their anti-tumor spectrum is narrow; and they activate the S6K / IRS1 / PI3K negative feedback pathway, which leads to the weakening of their own anti-tumor effect. The second generation of mTOR inhibitors are ATP competitive inhibitors, which inhibit mTORC1 and mTORC2 at the same time, and can effectively avoid the release of negative feedback pathways such as S6K / IRS1 / PI3K. Several compounds have entered clinical research. Studies have found that tumor cells can mediate their resistance to first-generation and second-generation drugs through mutations in the FRB region and ATP binding pocket of the mTOR protein. At the same time, PAM pathway inhibitors including mTOR inhibitors can activate related signal bypasses, which in turn lead to the occurrence of drug resistance.

[0008] Histone deacetylase (HDAC) can catalyze the deacetylation of histones and non-histone substrates, and regulate the acetylation balance in cells together with histone acetyltransferase (HAT). As an important anti-tumor target in epigenetics, HDAC can inhibit the transcription of tumor suppressor genes such as P53 and PTEN, and can regulate intracellular signal transduction, thereby affecting the proliferation, apoptosis and angiogenesis of tumor cells. The HDAC inhibitors (HDACis) that have been approved for marketing, Vorinostat (SAHA), Belinostat, Panobinostat and Chidamide, are used to treat cutaneous T-lymphocytic neoplasms, peripheral T-lymphocytic neoplasms and multiple myeloma, which fully confirms the application value of HDAC as an anti-tumor target.

[0009]

[0010] Based on the closely related carcinogenic mechanisms, multi-pathway and multi-way intervention in the occurrence and development of tumors through combination therapy or the use of single-molecule multi-target drugs is a feasible strategy to obtain synergistic efficacy and weaken drug resistance. In 2020, Filip Janku's research group explored the efficacy of the HDAC inhibitor vorinostat combined with the mTOR inhibitor everolimus (V+E). The results showed that the combined inhibition of HDAC and mTOR had an encouraging effect on patients with relapsed / refractory Hodgkin's lymphoma, which provided a theoretical basis for the research and development of mTOR / HDAC multi-target inhibitors. Compared with combination therapy, the use of single-molecule multi-target drugs can not only produce synergistic effects and enhance the therapeutic effect, but also circumvent drug-drug interactions, simplify pharmacokinetic properties, reduce off-target effects, and improve patient compliance. At present, PI3Ks / HDAC single-molecule dual-target inhibitors include CUDC-907 and 14w. Among them, the pro-apoptotic activity and anti-tumor growth of CUDC-907 have been shown to be more effective than single-target PI3K or HDAC inhibitors in various implanted and cultured cancer cell lines of T-cell lymphoma and leukemia. Phase I clinical studies have shown that dual HDAC / PI3K inhibition of CUDC-907 is well tolerated with a good toxicity profile, which is slightly better than or equivalent to FDA-approved single-target PI3K and HDAC drugs.

[0011]

[0012] PI3Ks / mTOR / HDAC multi-target inhibitors are not only expected to achieve better efficacy than mTOR inhibitors and weaken their drug resistance, but also to circumvent the defects of the combination therapy of PI3K inhibitors, mTOR inhibitors, and HDAC inhibitors. Therefore, it is of great significance to develop single-molecule multi-target inhibitors of PI3Ks / mTOR / HDAC. Summary of the invention

[0013] In view of the deficiencies of the prior art, the present invention proposes a diaryl urea PI3K / mTOR / HDAC multi-target inhibitor and a pharmaceutical composition and application thereof, so as to screen out compounds with better effectiveness and selectivity for the preparation of drugs for treating cancer.

[0014] This is achieved specifically through the following technical solutions:

[0015] The first object of the present invention is to provide a diaryl urea PI3K / mTOR / HDAC multi-target inhibitor, which includes a substituted triazine compound having a general formula (I), or a stereoisomer, a hydrate or a pharmaceutically acceptable salt thereof, wherein the general formula (I) has the following structure:

[0016]

[0017] Wherein, in the general formula (I), R1 and R2 are selected from H, C1-C4 alkyl, C1-C4 alkyl containing one or more substituents, C1-C4 alkoxy, C1-C4 alkoxy containing one or more substituents, C1-C4 alkyl acyl, C1-C4 alkyl sulfonyl, C3-C6 heterocyclic group, C3-C6 heterocyclic group containing one or more substituents, C4-C8 fused heterobicyclic group or C4-C8 fused heterobicyclic group containing one or more substituents; X is selected from the following structures:

[0018] n=1 or 0.

[0019] Further, the substituents on R1 or R2 are selected from fluorine, chlorine, bromine, iodine, hydroxyl, amino, cyano, C1-C4 alkyl, halogenated C1-C4 alkyl, hydroxyl C1-C4 alkyl, C1-C4 alkoxy, halogenated C1-C4 alkoxy, hydroxyl C1-C4 alkoxy or C1-C4 alkoxy C1-C4 alkyl.

[0020] Furthermore, in the general formula (I), R1 and R2 are independently selected from the following structures:

[0021]

[0022] Furthermore, the hydrogen attached to the carbon in the general formula (I) can be replaced by deuterium, an isotope of hydrogen; for example, the alkyl group can be replaced by a deuterated alkyl group, the alkoxy group can be replaced by a deuterated epoxy group, the benzene ring can be replaced by a deuterated benzene ring, and the aromatic ring can be replaced by a deuterated aromatic ring.

[0023] A pharmaceutically acceptable salt refers to a salt form in which the basic group in the parent compound is converted. Pharmaceutically acceptable salts include, but are not limited to, inorganic or organic acid salts of basic groups such as amine (amino) groups. The pharmaceutically acceptable salts of the present invention can be synthesized from the parent compound, i.e., the basic group in the parent compound is reacted with 1-4 equivalents of acid in a solvent system. Suitable salts are listed in Remington's Pharmaceutical Sciences, 17th ed., Mack Publishing Company, Easton, Pa., 1985, p. 1418 and Journal of Pharmaceutical Science, 66, 2 (1977).

[0024] The basic groups of the compounds of the present invention can form salts with acids, and examples of these acid salts include: salts formed with inorganic acids, especially hydrohalic acids (such as hydrochloric acid, hydrobromic acid, hydroiodic acid), nitric acid, sulfuric acid, phosphoric acid, carbonic acid, etc.; salts formed with lower alkyl sulfonic acids, such as methanesulfonic acid and trifluoromethanesulfonic acid; salts formed with aryl sulfonic acids, such as benzenesulfonic acid or p-toluenesulfonic acid; salts formed with organic acids, such as acetic acid, fumaric acid, tartaric acid, oxalic acid, citric acid, maleic acid, malic acid or succinic acid; and salts formed with amino acids, such as aspartic acid or glutamic acid.

[0025] The compounds of the present invention and pharmaceutically acceptable salts also include solvates or hydrates. In general, the solvates or hydrates are equivalent to non-solvated or non-hydrated forms and are included within the scope of the present invention. Some compounds of the present invention may exist in polycrystalline or amorphous forms. In general, all physical forms have equivalent uses and are included within the scope of the present invention.

[0026] In addition, unless otherwise indicated, the structural formula of the substituted triazine compound in the diaryl urea PI3Ks / mTOR / HDAC multi-target inhibitor of the present invention includes all isomeric forms (such as enantiomers, diastereomers, and geometric isomers (or conformational isomers)): for example, R, S configurations containing asymmetric centers, (Z), (E) isomers of double bonds, and (Z), (E) conformational isomers. Therefore, single stereochemical isomers of the compounds of the present invention or mixtures of their enantiomers, diastereomers, or geometric isomers (or conformational isomers) all fall within the scope of the present invention.

[0027] Unless otherwise indicated, all tautomeric forms of the pyrimidine compounds in the diaryl urea PI3Ks / mTOR / HDAC multi-target inhibitors of the present invention are included within the scope of the present invention. In addition, unless otherwise indicated, the structural formula of the compounds described in the present invention includes one or more enriched isotopes of different atoms.

[0028] The diaryl urea PI3K / mTOR / HDAC multi-target inhibitor comprises one of the following structures:

[0029] (1) 4-(3-(4-(4,6-dimorpholinyl-1,3,5-triazine-2-yl))phenyl)ureido)-N-hydroxybenzamide;

[0030] (2) (S)-N-hydroxy-4-(3-(4-(4-(3-methylmorpholinyl))-6-morpholin-1,3,5-triazinyl-2-yl)phenyl)ureido))benzamide;

[0031] (3) 4-(3-(4-(4-((1R,5S))-3-oxa-azabicyclo[3.2.1]octan-8-yl)-6-morpholin-1,3,5-triazin-2-yl)phenyl)ureido)-N-hydroxybenzamide;

[0032] (4) 4-(3-(4-(4,6-((1R,5S))-3-oxa-azabicyclo[3.2.1]octan-8-yl)-1,3,5-triazin-2-yl)phenyl)ureido)-N-hydroxybenzamide;

[0033] (5) 4-(3-(4-(4-(3S,5R)-3,5-dimethylmorpholine)-6-morpholine-1,3,5-triazine-2-yl)phenyl)ureido)-N-hydroxybenzamide;

[0034] (6) N-hydroxy-4-(3-(4-(4-(R)-3-methylmorpholine ring)-6-((S)-3-methylmorpholine ring)-1,3,5-triazine ring-2-yl)phenyl)ureido)benzamide;

[0035] (7) (R)-N-hydroxy-4-((3-(4-(4-(3-methylmorpholinyl))-6-morpholinyl-1,3,5-triazinyl)phenyl)ureido)methyl)benzamide;

[0036] (8) (R)-N-(2-amino-4-fluorophenyl)-4-(3-(4-(4-(3-methylmorpholinyl))-6-morpholino-1,3,5-triazin-2-yl)phenyl)ureido)benzamide;

[0037] (9) N-hydroxy-4-(3-(4-(4-(2-methylmorpholine)))-6-morpholine-1,3,5-triazine-2-yl)phenyl)ureido)benzamide;

[0038] (10) (R,E)-N-hydroxy-3-(4-(3-(4-(3-methylmorpholine))-6-morpholine-1,3,5-triazine-2-yl)phenyl)ureido)phenyl)acrylamide;

[0039] (11) (R)-N-hydroxy-4-(3-(4-(4-(3-methylmorpholinyl))-6-morpholino-1,3,5-triazin-2-yl)phenyl)ureido)benzamide;

[0040] (12) (E)-3-(4-(3-(4-(4,6-dimorpholinyl-1,3,5-triazinyl))phenyl)ureido)phenyl)-N-hydroxymethyl acrylamide;

[0041] (13) 4-(3-(4-(4-(2,6-dimethylmorpholinyl))-6-morpholinyl-1,3,5-triazine-2-yl)phenyl)ureido)-N-hydroxybenzamide;

[0042] (14) 4-(3-(4-(4-dimethylamino)piperidin-1-yl)-6-morpholino-1,3,5-triazine-2-yl)phenyl)ureido)-N-hydroxybenzamide;

[0043] (15) N-(2-amino-4-fluorophenyl)-4-(3-(4-(4,6-dimorpholinyl-1,3,5-triazinyl))phenyl)ureido)benzamide.

[0044] The preparation method of the diaryl urea PI3K / mTOR / HDAC multi-target inhibitor is as follows: cyanuric chloride (a) is used as a raw material, and the two chlorine atoms of the raw material (a) are subjected to a substitution reaction with morpholine or other nitrogen-containing heterocycles to introduce the R1 group to obtain an intermediate (b); on the basis of the intermediate (b), the R2 group is introduced through a nucleophilic substitution reaction to obtain an intermediate (c); the intermediate (c) and 4-aminophenylboronic acid pinacol ester undergo a Suzuki coupling reaction to obtain an intermediate (d); the amino end of the intermediate (d) is connected with 4-aminobenzoic acid methyl ester through a urea group to obtain an intermediate (e); the intermediate (e) undergoes a substitution reaction with hydroxylamine to obtain the target compound (f). The preparation route is as follows:

[0045]

[0046] In the above preparation steps, the substituents R1 and R2 in formulae a to f have the same meanings as the substituents R1 and R2 in the general formula (I).

[0047] The preparation method of the diaryl urea PI3K / mTOR / HDAC multi-target inhibitor is to hydrolyze the intermediate (e) obtained above to obtain the intermediate (f); the intermediate (f) is subjected to amide condensation to obtain the target compound (g); the preparation route is as follows:

[0048]

[0049] In the above preparation steps, the substituents R1 and R2 in formulae a to g have the same definitions as the substituents R1 and R2 in the general formula (I).

[0050] The preparation method of the diaryl urea PI3K / mTOR / HDAC multi-target inhibitor is to condense the intermediate (d) obtained above with 4-aminophenyl acrylate in the presence of triphosgene to obtain the intermediate (e); the intermediate (e) is subjected to a substitution reaction with hydroxylamine to obtain the target compound (f); the preparation route is as follows:

[0051]

[0052] In the above preparation steps, the substituents R1 and R2 in formulae a to g have the same definitions as the substituents R1 and R2 in the general formula (I).

[0053] The second object of the present invention is to provide a pharmaceutical composition comprising at least one pharmaceutically acceptable excipient, adjuvant or carrier, and an effective therapeutic dose of at least one of the aforementioned diaryl urea PI3Ks / mTOR / HDAC multi-target inhibitors.

[0054] The third object of the present invention is to provide a use of the aforementioned diaryl urea PI3Ks / mTOR / HDAC multi-target inhibitor or the aforementioned pharmaceutical composition in the preparation of a drug for preventing and / or treating and / or assisting in the treatment of proliferative diseases, metabolic diseases, nervous system diseases and tuberous sclerosis caused by overactivation of mTOR kinase and HDAC.

[0055] The proliferative diseases include colorectal cancer, gastric cancer, breast cancer, lung cancer, liver cancer, prostate cancer, pancreatic cancer, thyroid cancer, bladder cancer, kidney cancer, brain cancer, cervical cancer, cancer of the CNS, malignant glioma, myeloproliferative disease, blood cancer or lymphoma.

[0056] The fourth object of the present invention is to provide a use of the aforementioned diaryl urea PI3Ks / mTOR / HDAC multi-target inhibitor or the aforementioned pharmaceutical composition in the preparation of a drug for inhibiting cancer cell growth in vitro.

[0057] The terms used herein have the following meanings:

[0058] The term "alkyl" refers to a saturated straight or branched monovalent hydrocarbon group containing 1 to 20 carbon atoms, wherein the alkyl group may be independently optionally substituted with one or more substituents described herein. In some embodiments, the alkyl group contains 1 to 10 carbon atoms, in other embodiments, the alkyl group contains 1 to 8 carbon atoms, in other embodiments, the alkyl group contains 1 to 6 carbon atoms, and in other embodiments, the alkyl group contains 1 to 4 carbon atoms. Further examples of alkyl groups include, but are not limited to, methyl (Me, -CH3), ethyl (Et, -CH2CH3), n-propyl (n-Pr, -CH2CH2CH3), isopropyl (i-Pr, -CH(CH3)2), n-butyl (n-Bu, -CH2CH2CH2CH3), isobutyl (i-Bu, -CH2CH(CH3)2), sec-butyl (s-Bu, -CH(CH3)CH2CH3), tert-butyl (t-Bu, -C(CH3)3), and the like. The term "alkyl" and its prefix "alkane" as used herein include both straight and branched saturated carbon chains.

[0059] The term "alkoxy" refers to an alkyl group having the same meaning as the aforementioned "alkyl" and is formed by an oxygen atom attached to the main carbon chain of the "alkyl" group.

[0060] The term "haloalkyl" or "haloalkoxy" refers to a situation where an "alkyl" or "alkoxy" group can be substituted by one or more identical or different halogen atoms. The alkyl and alkoxy groups have the meanings as described above in the present invention, and such examples include but are not limited to trifluoromethyl, trifluoromethoxy, etc.

[0061] The term "hydroxyalkyl" or "hydroxyalkoxy" refers to the case where an "alkyl" or "alkoxy" group may be substituted by one or more hydroxyl groups. The "alkyl" and "alkoxy" groups have the meanings as described above in the present invention, and such examples include but are not limited to hydroxymethyl, 1-hydroxyethyl, hydroxypropyl, 1,2-dihydroxypropyl, hydroxymethoxy, 1-hydroxyethoxy, and the like.

[0062] The term "halogen", "halogen atom" or "halogen atom" includes fluorine, chlorine, bromine, iodine.

[0063] The term "heterocyclyl" may be a carbon group or a heteroatom group. "Heterocyclyl" also includes groups formed by the combination of a heterocyclic group with a saturated or partially unsaturated ring or heterocyclic ring. Examples of heterocyclic rings include, but are not limited to, pyrrolidinyl, tetrahydrofuranyl, dihydrofuranyl, tetrahydrothiophenyl, tetrahydropyranyl, dihydropyranyl, tetrahydrothiopyranyl, piperidinyl, thioxanyl, azetidinyl, oxetanyl, thietanyl, piperidinyl, homopiperidinyl, glycidyl, azepanyl, oxetanyl, thietanyl, N-morpholinyl, 2-morpholinyl, 3-morpholinyl, thiomorpholinyl, N-piperazinyl, 2-piperazinyl, 3-piperazinyl, homopiperazinyl, 4-methoxy-piperidin-1-yl, 1,2 , 3,6-tetrahydropyridin-1-yl, oxazepinyl, diazepinyl, thiazepinyl, pyrrolin-1-yl, 2-pyrrolinyl, 3-pyrrolinyl, indolinyl, 2H-pyranyl, 4H-pyranyl, dioxane, 1,3-dioxolane, pyrazolinyl, dithianyl, dithiolanyl, dihydrothiophenyl, pyrazolidinylimidazolinyl, imidazolidinyl, 1,2,3,4-tetrahydroisoquinolinyl, 1,2,6-thiadiazinane, 1,1-dioxo-2-yl, quinolizinyl and N-pyridylurea. And the heterocyclic group may be substituted or unsubstituted, wherein the substituent may be but is not limited to oxo (=O), hydroxyl, amino, halogen, cyano, heteroaryl, alkoxy, alkylamino, alkyl, alkenyl, alkynyl, heterocyclic group, thiol, nitro, aryloxy, hydroxy-substituted alkoxy, hydroxy-substituted alkyl-C (=O), alkyl-C (=O), carboxyalkoxy, etc.

[0064] The term "fused bicyclic ring", "fused ring", "fused bicyclic radical" or "fused cyclic radical" refers to a saturated or unsaturated fused ring system, and refers to a non-aromatic bicyclic ring system. Such a system may contain independent or conjugated unsaturation, but its core structure does not contain aromatic rings or heteroaromatic rings (although aromatic rings can be substituted thereon). Each ring in the fused bicyclic ring is either carbocyclic or heteroalicyclic, and such examples include but are not limited to 2,3,3a,4,7,7a-hexahydro-1H-indenyl, 7-azabicyclo[2.2.1]heptyl, fused bicyclo[3.3.0]octyl, fused bicyclo[3.1.0]hexyl, 1,2,3,4,4a,5,8,8a-octahydronaphthyl, all of which are included in the fused bicyclic ring system. And the fused bicyclic group can be substituted or unsubstituted, wherein the substituent can be but is not limited to halogen, hydroxyl, amino, cyano, aryl, heteroaryl, alkoxy, alkyl, alkenyl, alkynyl, heterocyclic group, thiol, nitro, aryloxy and the like.

[0065] The term "fused heterobicyclic group" refers to a saturated or unsaturated fused ring system, and relates to a non-aromatic bicyclic ring system. Such a system may contain independent or conjugated unsaturated states, but its core structure does not contain aromatic rings or aromatic heterocyclic rings (but aromatics can be substituents thereon). And at least one ring system contains one or more heteroatoms, wherein each ring system contains a 3-7-membered ring, that is, 1-6 carbon atoms and 1-3 heteroatoms selected from N, O, P, S, where S or P is optionally substituted by one or more oxygen atoms to obtain groups such as SO, SO2, PO, PO2, such examples include but are not limited to hexahydro-2H-[1,4]dioxo[2,3-c]pyrrolyl, etc. And the fused heterobicyclic group may be substituted or unsubstituted, wherein the substituents may be but are not limited to halogen, hydroxyl, amino, cyano, aryl, heteroaryl, alkoxy, alkyl, alkenyl, alkynyl, heterocyclyl, thiol, nitro, aryloxy, etc.

[0066] The term "therapeutically effective dose" refers to an amount of a compound of the formula sufficient to be effective for treatment when administered to a mammal in need of such treatment. The therapeutically effective amount will vary depending on the specific activity of the therapeutic agent used, the age, physiological condition, presence of other disease states, and nutritional status of the patient. In addition, other drug treatments that the patient may be receiving will affect the determination of the therapeutically effective amount of the therapeutic agent to be administered.

[0067] The term "treat" means any treatment of a disease in a mammal, including: (i) preventing the disease, i.e., causing clinical symptoms of the disease not to develop; (ii) inhibiting the disease, i.e., arresting the development of clinical symptoms; and / or (iii) palliating the disease, i.e., causing regression of clinical symptoms.

[0068] The term "pharmaceutically acceptable excipient, adjuvant or carrier" includes any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like. The use of such media and agents for pharmaceutically active substances is well known in the art. Unless any conventional media or agent is incompatible with the active ingredient, its use in the therapeutic composition is contemplated. Supplementary active ingredients may also be incorporated into the composition.

[0069] The composition is preferably formulated in a unit dosage form. The term "unit dosage form" refers to a physically discrete unit suitable for use as a single dose for administration to human subjects and other mammals, each unit containing a predetermined amount of active substance calculated to produce the required therapeutically effective amount and a related suitable pharmaceutical excipient (such as a tablet, capsule, ampoule). The pyrimidine compound in the diaryl urea PI3Ks / mTOR / HDAC multi-target inhibitor is effective over a wide range of doses and is generally administered in an effective amount of drug. Preferably, for oral administration, each dosage unit contains 10 mg to 2 g of the pyrimidine compound in the diaryl urea PI3Ks / mTOR / HDAC multi-target inhibitor, more preferably 10 to 700 mg, and for parenteral administration, preferably 10 to 700 mg of the pyrimidine compound in the diaryl urea PI3K / mTOR / HDAC multi-target inhibitor, more preferably about 50 to 200 mg. However, it should be understood that the amount of pyrimidine compound in the diaryl urea PI3K / mTOR / HDAC multi-target inhibitor actually administered will be determined by the physician based on relevant circumstances, including the condition to be treated, the selected route of administration, the actual compound administered and its relative activity, the age, weight, and response of each patient, the severity of the patient's symptoms, etc.

[0070] In order to prepare solid compositions such as tablets, the main active ingredient is mixed with a pharmaceutical excipient (or carrier) to form a solid preformulation composition, which contains a uniform mixture of the compound of the present invention. When these preformulation compositions are referred to as uniform, it means that the active ingredient is evenly dispersed throughout the composition, so that the composition can be easily subdivided into equally effective unit dosage forms such as tablets, pills and capsules.

[0071] Tablet or pill of the present invention can be coated or otherwise compounded to provide a dosage form with a prolonged effect advantage, or protect tablet or pill from the effect of acidic conditions in the stomach. For example, tablet or pill can include inner dosage and outer dosage components, the latter having the form of the outer skin on the former. Two kinds of components can be separated with an enteric layer, wherein the enteric layer is used to prevent the disintegration in the stomach and allows the complete entry of the inner component into the duodenum or delayed release. Various materials can be used for such enteric layers or coatings, and the above-mentioned materials include many macromolecular acids and macromolecular acids and such materials as shellac, hexadecanol and cellulose acetate mixture.

[0072] Compositions for inhalation or insufflation include solutions and suspensions in pharmaceutically acceptable aqueous or organic solvents, or mixtures thereof, and powders. Liquid or solid compositions may contain suitable pharmaceutical excipients as described above. Preferably, these compositions are administered by oral or nasal respiratory route to obtain local or systemic effects. Compositions in preferred pharmaceutically acceptable solvents may be atomized by the use of inert gases. Atomized solutions may be inhaled directly from an atomizing device, or the atomizing device may be connected to a mask tent, or an intermittent positive pressure breathing machine. Solutions, suspensions, or powder compositions may be administered by a device that delivers the dosage form in an appropriate manner, preferably by oral or nasal route.

[0073] Beneficial effects:

[0074] The compound of the present invention is a single-molecule multi-target inhibitor, which has the characteristics of low cost, good efficacy and low toxicity, and the intermediate product yield in the synthesis process is high, which reduces resource waste and is conducive to reducing costs.

[0075] The compound of the present invention has the characteristics of effectively inhibiting the PI3K / mTOR signaling pathway and HDAC protein, and has excellent mTOR inhibitory activity.

[0076] The compound of the present invention has a novel structure, significant anti-tumor activity, and a small dosage, and can be potentially used in the treatment of related tumors.

[0077] The compounds of the present invention have positive and predictable clinical application value in anti-proliferative diseases, especially anti-tumor, and have good development prospects. DETAILED DESCRIPTION

[0078] The technical solution of the present invention is further defined below in conjunction with specific implementation methods, but the scope of protection required is not limited to the description.

[0079] In the examples described below, all temperatures are in degrees Celsius unless otherwise indicated. Reagents were purchased from commercial suppliers such as AlfaAesar Chemical Company, Bailingwei Technology Co., Ltd., Aladdin Reagent Co., Ltd., Beijing Coupling Technology Co., Ltd., etc., and were used without further purification unless otherwise indicated. General reagents were purchased from Shantou Xilong Chemical Plant, Guangzhou Chemical Reagent Factory, Tianjin Zhiyuan Chemical Reagent Co., Ltd., and Qingdao Ocean Chemical Plant, etc.

[0080] In the embodiments described below, the chromatographic column uses a silica gel column, and the silica gel (200-300 mesh) is purchased from Qingdao Ocean Chemical Plant. The nuclear magnetic resonance spectrum uses CDCl3 or DMSO-d6 as solvent (in ppm), and TMS (0 ppm) or chloroform (7.26 ppm) is used as a reference standard. When multiple peaks appear, the following abbreviations will be used: s (singlet, single peak), d (doublet, double peak), t (triplet, triple peak), m (multiplet, multiple peak), br (broadened, broad peak), dd (doublet of doublets, quartet), dt (doublet of triplets, double triplet). Coupling constants are expressed in Hertz (Hz).

[0081] The low-resolution mass spectrometry (MS) data in the examples described below were measured by an Agilent 6120 series LC-MS spectrometer equipped with a G1311B quaternary pump and a G1316BTCC (column temperature maintained at 30°C), a G1329B autosampler and a G1315C DAD detector were used for analysis, and an ESI source was applied to the LC-MS spectrometer.

[0082] In the examples described below, the injection volume was determined by the sample concentration; the flow rate was 0.5 mL / min; the HPLC peaks were recorded and read by UV-Vis wavelengths at 210 nm and 254 nm. The mobile phase was isopropanol / n-hexane (40:60).

[0083] For the convenience of description, some raw materials in the embodiments described below will be described by their abbreviations, which are compared with their full names as follows: DCM is CH2Cl2, i.e. dichloromethane; CDC13 is deuterated chloroform; PE is petroleum ether; EtOAc and EA are both ethyl acetate; MeOH and CH3OH are both methanol; Pd(PPh3)4 is tetrakistriphenylphosphine palladium; DIPEA is N,N-diisopropylethylamine; DMSO-d6 is hexadeuterated dimethyl sulfoxide; DME is ethylene glycol dimethyl ether; Na2SO4 is sodium sulfate.

[0084] Example 1: (S)-N-hydroxy 4-(3-(4-(4-(3-methylmorpholinyl))-6-morpholin-1,3,5-triazine ring-2-yl)phenyl)urea))benzamide, the synthesis steps are as follows:

[0085] Step 1: Synthesis of 4-(4,6-dichloro-1,3,5-triazine-2-yl)morpholine, structural formula:

[0086] Weigh the raw material cyanuric chloride (10.84mmol) and put it into a 100mL double-necked bottle, add DCM to dissolve, then add DIPEA (10.84mmol), circulate the vacuum nitrogen three times, cool at low temperature -78℃ for 10min, slowly add morpholine (10.84mmol) through a constant pressure dropping funnel and keep the low temperature for 1h, monitor the reaction completion by TLC, directly vacuum concentrate the reaction solution, mix with silica gel, and purify by silica gel column chromatography (PE:EA=5:1) to obtain the product. White powder, yield: 78.13%. HRMS (ESI) calcd.forC7H8C l2 N4O[M+H] + :235.0153,found:235.0159;

[0087] Step 2: Synthesis of (S)-4-(4-chloro-6-morpholino-1,3,5-triazine-2-yl)-3-methylmorpholine, structural formula:

[0088]

[0089] 4-(4,6-dichloro-1,3,5-triazine-2-yl)morpholine (1.08mmol), DIPEA (1.13mmol), and morpholine (1.08mmol) were added to a 50mL double-necked bottle in sequence, dissolved with DCM, replaced with nitrogen, and reacted at 0°C for 3h. After the reaction of the raw materials was completed by TLC monitoring, the nitrogen was removed, water and DCM (3×20mL) were added to the reaction mixture for extraction, the organic layers were combined, and then the organic phase was dried with anhydrous Na2SO4, the solvent was evaporated, and purified by silica gel column chromatography (PE:EA=10:1). White solid, yield: 87.5%. NMR data is 1 H NMR (400MHz, CDCl3) δ4.67(s,1H),4.33(d,J=14.8Hz,1H),3.94(dd,J=11.5,3.7Hz,1H),3.79(t,J=5.7Hz,4H),3.71(t,J=4.7Hz,5 H),3.63(dd,J=11.5,3.3Hz,1H),3.48(td,J=11.9,3.0Hz,1H),3.25(ddd,J=13.7,12.3,3.8Hz,1H),1.30(d,J=6.9Hz,3H).HRMS(ES I)calcd.for C 12 H 18 ClN5O2[M+H] + :300.1227,found:300.1221;

[0090] Step 3: Synthesis of (S)-4-(4-(3-methylmorpholino)-6-morpholino-1,3,5-triazine-2-yl)aniline, structural formula:

[0091] To a 50mL single-necked bottle, (S)-4-(4-chloro-6-morpholino-1,3,5-triazine-2-yl)-3-methylmorpholine (2.13mmol), 4-aminophenylboronic acid pinacol ester (2.34mmol), tetrakis(triphenylphosphine)palladium (0.1mmol), potassium carbonate (5.32mmol) were added in sequence, and dissolved with DME (10mL) and water (2mL). The reaction was replaced with nitrogen, refluxed at 100°C for 12h, and the reaction was completed by TLC tracking. It was cooled to room temperature, EA (50mL×3) and water were added for extraction, the organic phases were combined, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by silica gel column chromatography (DCM:MeOH=50:1). Yellow solid, yield: 88.6%. NMR data is 1 H NMR (600MHz, DMSO-d6) δ8.05(d,J=8.6Hz,2H),6.58(d,J=8.6Hz,2H),5.71(s,2H),4.75(s,1H),4.37(s,1H),3.90(d ,J=11.0Hz,1H),3.71(s,5H),3.64(s,4H),3.57(s,1H),3.41(d,J=11.1Hz,1H),3.15(s,1H),1.21(d,J=6.8Hz,3H); 13 C NMR(151MHz,DMSO-d6)δ169.9,165.1,164.7,152.7,130.2,123.9,113.2,70.8,66.7,66.5,46.0,14.4.HRMS(ESI)calcd.for C 18 H 24 N6O2[M+H] + :357.2039,found:357.2039.

[0092] Step 4: Synthesis of methyl (S)-4-(3-(4-(4-(3-methylmorpholinyl))-6-morpholin-1,3,5-triazine-2-yl)phenyl)ureido)benzoate, the structural formula is as follows;

[0093]

[0094] (S)-4-(4-(3-methylmorpholino)-6-morpholino-1,3,5-triazine-2-yl)aniline (0.66mmol) was added to a 50mL single-necked bottle and dissolved with 1,4-dioxane (5mL). The mixture was cooled to 0-4°C and triphosgene (0.66mmol) was added. The mixture was refluxed at 110°C for 12h. The reaction was completed by TLC. The solvent was removed from the reaction solution to obtain an isocyanate intermediate. The residue was dissolved in DCM (5mL), and the intermediate 4-aminobenzoic acid methyl ester (0.66mmol) was added and reacted at room temperature for 12h. The reaction was completed by TLC tracking and purified by silica gel column chromatography (DCM / MeOH=20:1). White solid, yield: 24%. HRMS (ESI) calcd.for C 27 H 31 N7O5[M+H] + :533.2387,found:532.2380.

[0095] Step 5: Synthesis of (S)-N-hydroxy 4-(3-(4-(4-(3-methylmorpholinyl))-6-morpholino-1,3,5-triazine-2-yl)phenyl)ureido)benzamide, structural formula;

[0096]

[0097] Add (S)-methyl 4-(3-(4-(4-(3-methylmorpholinyl))-6-morpholin-1,3,5-triazine ring-2-yl)phenyl)ureido)benzoate (0.66mmol) to a 50mL single-necked bottle, dissolve with methanol / water (10mL), add sodium hydroxide (6.6mmol) and hydroxylamine hydrochloride at room temperature, reflux at 40°C for 12h, and complete the reaction by TLC tracking. Remove the solvent from the reaction solution and purify it by silica gel column chromatography (DCM / MeOH=20:1). White solid, yield: 23%. 1 H NMR (400MHz, DMSO-d6) δ11.09(s,1H),9.12(s,1H),9.02(s,1H),8.92(s,1H),8.28(d,J= 8.3Hz,2H),7.71(d,J=8.3Hz,2H),7.55(dd,J=14.6,8.3Hz,4H),4.78(s,1H),4.40(s,1H) ,3.92(d,J=11.1Hz,2H),3.81(s,3H),3.72(d,J=11.4Hz,1H),3.66(t,J=5.6Hz,4H),3.58 (d,J=11.8Hz,1H),3.44(d,J=11.7Hz,1H),3.19(t,J=12.8Hz,1H),1.24(d,J=6.8Hz,3H); 13C NMR(100MHz,DMSO-d6)δ169.4,165.1,164.7,164.5,152.5,143.1,142.6,130.6,129.5,1 28.3,126.4,117.9,117.8,74.0,70.7,66.7,46.2,43.7,38.6,14.5.HRMS(ESI)m / z:calcd for C 26 H 31 N8O5[M+H] + 535.2417; Found 535.2421.

[0098] Example 2: 4-(3-(4-(4,6-dimorpholinyl-1,3,5-triazine ring-2-yl))phenyl)urea)-N-hydroxybenzamide, the structural formula is:

[0099] Synthesis method: replace the substituted heterocyclic fragment in step 2 of Example 1 with morpholine, and the other steps and operations are the same as in Example 1; white solid, yield: 17%. NMR data are 1 H NMR(400MHz,DMSO-d6)δ11.05(s,1H),9.18(d,J=18.1Hz,1H),9.08(s,1H),8.82(s,1H ),8.01-7.94(m,2H),7.75-7.67(m,4H),7.71-7.62(m,2H),3.77(m,8H),3.68(m,8H); 13 C NMR(100MHz,DMSO-d6)δ169.5,165.7,164.3,162.3,151.0,143.6,142.4,130.0,129.2,126.4,125.1,118.1,117.4,66.0,44.5.HRMS(ESI)m / z:calcd for C 26 H 31 N8O5[M+H] + 521.2261; Found 521.2259..

[0100] Example 3: 4-(3-(4-(4-((1R,5S))-3-oxa-azabicyclo[3.2.1]octane-8-yl)-6-morpholine-1,3,5-triazine-2-yl)phenyl)ureido)-N-hydroxybenzamide, the structural formula is as follows:

[0101]

[0102] Synthesis method: replace the substituted heterocyclic fragments in steps 1 and 2 of Example 1 with (1R,5S)-3-oxa-8-azabicyclo[3.2.1]octane, and the other steps and operations are the same as in Example 1; white solid, yield: 16%. NMR data are 1 H NMR (400MHz, DMSO-d6) δ11.06(s,1H),9.91(d,J=42.0Hz,2H),8.21(d,J=8.3Hz,2H),7.73(d,J=8.2Hz, 2H),7.64(d,J=8.3Hz,2H),7.54(d,J=8.2Hz,2H),4.87(s,2H),4.69(s,2H),3.65(s,8H),1.98(s,8H); 13 C NMR(100MHz,DMSO-d6)δ164.5,152.6,142.6,130.2,128.3,126.3,117.6,117.5,71.8,71.3,55.2,51.1,26.7,26.5.HRMS(ESI)m / z:calcd forC 29 H 33 N8O5[M+H] + 573.2574; Found 573.2570.

[0103] Example 4: 4-(3-(4-(4,6-((1R,5S))-3-oxa-azabicyclo[3.2.1]octane-8-yl)-1,3,5-triazine-2-yl)phenyl)urea)-N-hydroxybenzamide, the structural formula is as follows:

[0104]

[0105] Synthesis method: replace the substituted heterocyclic fragment in step 2 of Example 1 with (1R,5S)-3-oxa-8-azabicyclo[3.2.1]octane, and the other steps and operations are the same as in Example 1; white solid, yield: 14%. NMR data are 1 H NMR (400MHz, DMSO-d6) δ11.07(s,1H),9.75(d,J=18.1Hz,2H),8.29-8.21(m,2H),7.72(d,J=8.8Hz,2H),7.57(d d,J=22.0,8.8Hz,4H),4.84(s,1H),4.67(s,1H),3.85(d,J=15.9Hz,4H),3.70-3.59(m,8H),2.01-1.85(m,4H); 13C NMR (100MHz, DMSO-d6) δ168.4,164.5,164.0,152.7,143.7,142.7,129.8,128.3,126. 3,117.6,117.5,71.5.71.2,66.4,55.1,54.8,43.9,26.8,26.7.HRMS(ESI)m / z:calcd for C 26 H 31 N8O5[M+H] + 547.2417; Found 547.2410.

[0106] Example 5: (R)-N-hydroxy 4-(3-(4-(4-(3-methylmorpholinyl))-6-morpholinyl-1,3,5-triazine-2-yl)phenyl)ureido)benzamide, the structural formula is as follows:

[0107]

[0108] Synthesis method: replace the substituted heterocyclic fragment in step 2 of Example 1 with (R)-3-methylmorpholine, and the other steps and operations are the same as in Example 1; white solid, yield: 15%. NMR data are 1 H NMR (400MHz, DMSO-d6) δ11.09(s,1H),9.12(s,1H),9.02(s,1H),8.92(s,1H),8.28(d,J= 8.3Hz,2H),7.71(d,J=8.3Hz,2H),7.55(dd,J=14.6,8.3Hz,4H),4.78(s,1H),4.40(s,1H) ,3.92(d,J=11.1Hz,2H),3.81(s,3H),3.72(d,J=11.4Hz,1H),3.66(t,J=5.6Hz,4H),3.58 (d,J=11.8Hz,1H),3.44(d,J=11.7Hz,1H),3.19(t,J=12.8Hz,1H),1.24(d,J=6.8Hz,3H); 13 C NMR(100MHz,DMSO-d6)δ169.4,165.1,164.7,164.5,152.5,143.1,142.6,130.6,129.5,1 28.3,126.4,117.9,117.8,74.0,70.7,66.7,46.2,43.7,38.6,14.5.HRMS(ESI)m / z:calcd for C 26 H 31 N8O5[M+H] +535.2417; Found 535.2421.

[0109] Example 6: 4-(3-(4-(4-(3S,5R)-3,5-dimethylmorpholine)-6-morpholine ring-1,3,5-triazine ring-2-yl)phenyl)ureido)-N-hydroxybenzamide, the structural formula is as follows:

[0110]

[0111] Synthesis method: replace the substituted heterocyclic fragment in step 2 of Example 1 with 3,5-dimethylmorpholine, and the other steps and operations are the same as in Example 1; white solid, yield: 18%. NMR data is 1 HNMR(400MHz,DMSO-d6)δ11.04(s,1H),9.10(s,1H),9.01(s,1H),8.82(s,1H),8.01-7.94(m,2H),7.75-7.67( m,2H),7.71-7.62(m,4H),4.05(s,2H),3.88(s,1H),3.81-3.74(m,7H),3.69(s,2H),3.55(s,2H),1.21(s,6H); 13 C NMR(100MHz,DMSO-d6)δ168.9,166.9,164.6,164.1,152.3,143.0,139.6,132.4,13 0.4,128.4,125.1,119.1,118.5,72.6,65.8,54.2,46.5,17.7.HRMS(ESI)m / z:calcd for C 26 H 31 N8O5[M+H] + 549.2574; Found 549.2570.

[0112] Example 7: N-hydroxy-4-(3-(4-(4-(R)-3-methylmorpholine ring)-6-((S)-3-methylmorpholine ring)-1,3,5-triazine ring-2-yl)phenyl)ureido)benzamide, the structural formula is as follows:

[0113]

[0114] Synthesis method: replace the substituted heterocyclic fragments in steps 1 and 2 of Example 1 with (S)-3-methylmorpholine, and the other steps and operations are the same as in Example 1; white solid, yield: 19%. NMR data are 1H NMR(400MHz,DMSO-d6)δ11.06(s,1H),9.08(s,1H),9.03(s,1H),8.82(s, 1H),,8.01-7.94(m,2H),7.75-7.71(m,2H),7.71-7.62(m,4H),4.75-4.5 5(m,2H),4.41-4.21(m,2H),3.93(dd,J=11,3.7Hz,2H),3.73(d,J=12Hz, 2H),3.63(dd,J=12,3.3Hz,2H),3.48(td,J=12.3Hz,2H),3.29-3.18(m,2

[0115] H), 1.30 (d, J = 6.9 Hz, 6H); 13 C NMR(100MHz,DMSO-d6)δ169.2,16

[0116] 5.3,164.7,164.5,152.6,143.2,142.7,130.6,129.6,128.4,126.5,118.1,117.8,72.2,67.2,50.2,41.5,15.7.HRMS(ESI)m / z:calcd for C 26 H 31 N8O5[M+H] + 549.2574; Found 549.2582.

[0117] Example 8: N-hydroxy 4-(3-(4-(4-(2-methylmorpholine)))-6-morpholine ring-1,3,5-triazine ring-2-yl)phenyl)urea)benzamide, the structural formula is as follows:

[0118]

[0119] Synthesis method: replace the substituted heterocyclic fragment in step 2 of Example 1 with 2-methylmorpholine, and the other steps and operations are the same as in Example 1; white solid, yield: 14%. NMR data are 1H NMR(400MHz,DMSO-d6)δ11.06(s,1H),9.10(s,1H),9.02(s,1H),8.90(s,1H),8.22(d,J=8.3Hz,2 H),7.79(d,J=8.3Hz,2H),7.59(dd,J=14.6,8.3Hz,4H),4.01(ddd,J=11.6,3.2,1.2Hz,1H),3.86 -3.71(m,2H),3.66(t,J=5.6Hz,4H),3.47(d,J=11.6Hz,1H),3.41(dd,J=12.0,1.6Hz,1H),2.96- 2.83(m,4H),2.82(td,J=11.6,3.6Hz,1H),2.48(dd,J=11.6,10.4Hz,1H),1.25(d,J=6.4Hz,3H); 13 CNMR(100MHz,DMSO-d6)δ169.4,165.2,164.7,164.5,152.6,143.1,142.6,130.7,129.5,12 8.3,126.4,117.9,117.8,73.8,70.8,66.8,46.4,43.7,38.7,14.7.HRMS(ESI)m / z:calcdfor C 26 H 31 N8O5[M+H] + 535.2417; Found 535.2422.

[0120] Example 9: 4-(3-(4-(4-(2,6-dimethylmorpholinyl))-6-morpholinyl-1,3,5-triazine-2-yl)phenyl)ureido)-N-hydroxybenzamide, the structural formula is as follows:

[0121]

[0122] Synthesis method: replace the substituted heterocyclic fragment in step 2 of Example 1 with 2,6-dimethylmorpholine, and the other steps and operations are the same as in Example 1; white solid, yield: 21%. NMR data is 1H NMR (400MHz, DMSO-d6) δ11.02(s,1H),9.10(s,1H),9.01(s,1H),8.94(s,1H),8.26(d,J=8.2Hz,2H),7.70(d,J=8.1Hz,2H ),7.50(dd,J=14.3,8.0Hz,4H),4.44-4.26(m,2H),3.75-3.69(m,10H),2.67(t,J=12Hz2H),3.57(s,2H),1.25(d,J=6.0Hz 6H); 13 C NMR(100MHz,DMSO-d6)δ169.5,165.1,164.8,164.5,152.6,143.2,142.7,130.7,12 9.5,128.2,126.4,117.9,117.8,71.4,65.8,54.4,46.5,18.8.HRMS(ESI)m / z:calcd for C 27 H 33 N8O5[M+H] + 549.2574; Found 549.2578.

[0123] Example 10: 4-(3-(4-(4-dimethylamino)piperidin-1-yl)-6-morpholine ring-1,3,5-triazine ring-2-yl)phenyl)urea)-N-hydroxybenzamide, the structural formula is as follows:

[0124]

[0125] Synthesis method: replace the substituted heterocyclic fragment in step 2 of Example 1 with N,N-dimethylpiperidin-4-amine, and the other steps and operations are the same as in Example 1; white solid, yield: 20%. NMR data is 1 H NMR(400MHz,DMSO-d6)δ10.94(s,1H),9.12(s,1H),9.05(s,1H),8.84(s,1H) ,8.26(d,J=8.6Hz,2H),7.74(d,J=8.3Hz,2H),7.50(dd,J=14.5,8.0Hz,4H),3 .82(m,4H),3.70(m,4H).3.68-3.77(m,2H),2.71(td,J=12.3,2.6Hz,2H),2.3 2(s,6H),2.24-2.31(m,1H),1.89-1.97(m,2H),1.64(qd,J=12.1,3.9Hz,2H); 13C NMR(100MHz,DMSO-d6)δ169.3,164.9,164.6,164.5,152.8,143.4,142.8,131.0,129.6 ,128.3,126.6,117.8,117.8,66.1,59.0,47.3,46.5,41.4,30.8.HRMS(ESI)m / z:calcd for C 27 H 33 N8O5[M+H] + 562.2890; Found 562.2894.

[0126] Example 11: (R,E)-N-hydroxy-3-(4-(3-(4-(3-methylmorpholine))-6-morpholine-1,3,5-triazine-2-yl)phenyl)urea)phenyl)acrylamide, the structural formula is as follows:

[0127]

[0128] The synthesis method comprises the following steps:

[0129] The substituted heterocyclic fragment in step 2 of Example 1 is replaced with (S)-3-methylmorpholine, and the other steps and operations are the same as step 3 of Example 1;

[0130] Step 4: Synthesis of methyl (R,E)-3-(4-(3-(4-(3-methylmorpholine))-6-morpholine-1,3,5-triazine-2-yl)phenyl)urea)phenyl)acrylate, structural formula:

[0131] Add the raw material (0.66mmol) to a 50mL single-necked bottle, dissolve it with 1,4-dioxane (5mL), cool the mixture to 0-4℃, add triphosgene (0.66mmol), reflux at 110℃ for 12h, and follow the reaction by TLC. Remove the solvent from the reaction solution to obtain the isocyanate intermediate, dissolve the residue with DCM (5mL), add the intermediate (E)-3-(4-aminophenyl) methyl acrylate (0.66mmol), and react at room temperature for 12h. Follow the reaction by TLC, and purify by silica gel column chromatography (DCM / MeOH=20:1). White solid, yield: 22%. White solid, yield: 20%. HRMS (ESI) m / z:calcd for C 29 H 34 N7O5[M+H] + 560.2621; Found 560.2619.

[0132] Step 5: Synthesis of (R,E)-N-hydroxy-3-(4-(3-(4-(3-methylmorpholine))-6-morpholine-1,3,5-triazine-2-yl)phenyl)ureido)phenyl)acrylamide, the structural formula is as follows;

[0133]

[0134] Synthesis method: refer to step 4 in Example 1. White solid, yield: 20%. NMR data is 1 H NMR (400MHz, DMSO-d6) δ10.67(s,1H),9.00(s,1H),8.82(s,1H),8.55(s,1H),8.22(d,J=8.2Hz,2H ),7.82(d,J=8.4Hz,2H),7.61-7.26(m,3H),6.89(d,J=8.8Hz,2H),6.36(d,J=15.8Hz,1H),4.80(s ,1H),4.44(s,1H),3.95(d,J=11.1Hz,2H),3.86(s,3H),3.78(d,J=11.5Hz,1H),3.70(t,J=5.4Hz, 4H), 3.61 (d, J = 11.5Hz, 1H), 3.45 (d, J = 11.5Hz, 1H), 3.19 (t, J = 12.7Hz, 1H), 1.24 (d, J = 6.6Hz, 3H); 13 C NMR(75MHz,DMSO-d6)δ169.4,164.6,164.3,163.2,154.6,152.5,141.2,138.2,132.5,128.3,12 8.1,120.2,118.0,116.5,114.0,74.2,70.2,65.8,46.5,43.8,39.0,16.45.HRMS(ESI)m / z:calcd for C 28 H 33 N8O5[M+H] + 561.2574; Found 561.2571.

[0135] Example 12: (E)-3-(4-(3-(4-(4,6-dimorpholinyl-1,3,5-triazine-2-yl))phenyl)urea)phenyl)-N-hydroxymethyl acrylamide, the structural formula is as follows:

[0136]

[0137] Synthesis method: replace the substituted heterocyclic fragment in step 2 of Example 1 with a morpholine ring, and the other steps and operations are the same as in Example 11, white solid, yield: 23%. NMR data are1 H NMR (400MHz, DMSO-d6) δ10.77(s,1H),9.05(s,1H),8.86(s,1H),8.50(s,1H),8.24(d,J=8.0Hz,2H),7.80(d ,J=8.1Hz,2H),7.64-7.24(m,3H),6.88(d,J=8.6Hz,2H),6.36(d,J=15.4Hz,1H),3.82(m,8H),3.70(m,8H); 13 CNMR(100MHz,DMSO-d6)δ169.3,164.4,164.2,163.2,154.5,152.5,141.1,138.3, 132.5,128.3,128.1,120.2,118.0,116.5,114.0,66.1,44.5.HRMS(ESI)m / z:calcd for C 27 H 31 N8O5[M+H] + 547.2417; Found 547.2410.

[0138] Example 13: N-(2-amino-4-fluorophenyl)-4-(3-(4-(4,6-dimorpholinyl-1,3,5-triazine-2-yl))phenyl)ureido)benzamide, the structural formula is as follows:

[0139]

[0140] The synthesis method comprises the following steps:

[0141] The substituted heterocyclic fragment in step 2 of Example 1 is replaced with a morpholine ring, and the other steps and operations are the same as steps 3 and 4 of Example 1;

[0142] Step 5: Synthesis of 4-(3-(4-(4,6-dimorpholinyl-1,3,5-triazinyl))phenyl)ureido)benzoic acid;

[0143]

[0144] Add the raw material (0.66mmol) to a 50mL single-necked bottle, dissolve it with methanol / tetrahydrofuran / water (10mL), add lithium hydroxide (6.6mmol) to the mixture at room temperature, reflux at 40℃ for 12h, and follow the reaction by TLC. Remove the solvent from the reaction solution, adjust the pH to acidic, and filter to obtain a white solid with a yield of 82%. NMR data are HRMS (ESI) m / z:calcd for C 27 H 31N8O5[MH]+504.1995; Found 504.1990.

[0145] Step 6: Synthesis of N-(2-amino-4-fluorophenyl)-4-(3-(4-(4,6-dimorpholinyl-1,3,5-triazinyl))phenyl)ureido)benzamide;

[0146]

[0147] Add the raw material (0.66 mmol) and 4-fluoro-o-phenylenediamine (0.66 mmol) to a 50 mL single-necked bottle, dissolve with DMF (10 mL), add DIPEA, HOBT, EDC to the mixture at room temperature, react overnight at room temperature, and follow the completion of the reaction by TLC. Remove the solvent from the reaction solution and purify by silica gel column chromatography (DCM / MeOH=20:1). Yield: 22%. NMR data is 1 H NMR (400MHz, DMSO-d6) δ9.47(s,1H),9.10(s,1H),9.02(s,1H),8.22(d,J=8.3Hz,2H),7.82(d,J=8.5Hz,2H),7.78(d,J=8.5Hz,2H),7.59(d d,J=14.6,8.3Hz,2H),7.12-7.08(m,1H),6.53(dd,J=3.0,11.5Hz,1H),6.35(dt,J=3.0,8.5Hz,1H),5.18(s,2H),3.82(m,8H),3.70(m,8H); 13 C NMR(100MHz,DMSO-d6)δ169.3,164.5,164.4,162.4,154.4,151.6,141.0,140.2,137.1,136.2,1 31.2,129.9,127.7,124.9,124.7,120.8,119.4,117.2,112.4,66.2,44.6.HRMS(ESI)m / z:calcd for C 31 H 33 FN9O4[M+H] + 614.2640; Found 614.2636.

[0148] Example 14: (R)-N-(2-amino-4-fluorophenyl)-4-(3-(4-(4-(3-methylmorpholinyl))-6-morpholinyl-1,3,5-triazine-2-yl)phenyl)ureido)benzamide, the structural formula is as follows:

[0149]

[0150] Synthesis method: replace the substituted heterocyclic fragment in step 2 of Example 1 with (S)-3-methylmorpholine, and the other steps and operations are the same as those of Example 13; white solid, yield: 21%. NMR data are 1 HNMR(400MHz,DMSO-d6)δ9.47(s,1H),9.10(s,1H),9.02(s,1H),8.22(d,J=8.3Hz,2H),7.82(d,J=8.5Hz,2H),7.7 8(d,J=8.5Hz,2H),7.59(dd,J=14.6,8.3Hz,2H),7.12-7.08(m,1H),6.53(dd,J=3.0,11.5Hz,1H),6.35(dt,J=3.0, 8.5Hz,1H),5.18(s,2H),4.78(s,1H),4.40(s,1H),3.92(d,J=11.3Hz,2H),3.86(s,3H),3.78(d,J=11.6Hz,1H),3. 70(t,J=5.8Hz,4H),3.61(d,J=11.7Hz,1H),3.45(d,J=11.7Hz,1H),3.19(t,J=12.8Hz,1H),1.24(d,J=6.8Hz,3H); 13 CNMR(100MHz,DMSO-d6)δ169.4,164.6,164.3,162.4,154.3,151.5,141.0,140.1,137.1,136.1,131.2,129.8,1 27.6,124.9,124.7,120.8,119.3,117.1,112.1,74.2,70.2,65.7,46.5,43.7,39.0,16.5.HRMS(ESI)m / z:calcd for C 32 H 35 FN9O4[M+H] + 628.2796; Found 628.2790.

[0151] Example 15: (R)-N-hydroxy-4-((3-(4-(4-(3-methylmorpholine))-6-morpholine-1,3,5-triazine-2-yl)phenyl)ureido)methyl)benzamide, the structural formula is as follows:

[0152]

[0153] The synthesis method comprises the following steps:

[0154] Steps 1 and 2: Step 1 and operation are the same as in Example 1, except that the substituted heterocyclic fragment in Step 2 of Example 1 is replaced by (S)-3-methylmorpholine;

[0155] Step 3: Step 3 and operation are the same as Step 3 of Example 1;

[0156] Step 4: Synthesis of (R)-methyl 4-((3-(4-(4-(3-methylmorpholine))-6-morpholinyl-1,3,5-triazine-2-yl)phenyl)ureido)methyl)benzoate, the structural formula is as follows:

[0157]

[0158] Add the raw material (0.66mmol) to a 50mL single-necked bottle, dissolve it with 1,4-dioxane (5mL), cool the mixture to 0-4℃, add triphosgene (0.66mmol), reflux at 110℃ for 12h, and follow the reaction by TLC. Remove the solvent from the reaction solution to obtain the isocyanate intermediate, dissolve the residue with DCM (5mL), add the intermediate 4-(aminomethyl)benzoic acid methyl ester (0.66mmol), and react at room temperature for 12h. Follow the reaction by TLC, and purify by silica gel column chromatography (DCM / MeOH=20:1). White solid, yield: 22%. White solid, yield: 20%.

[0159] Step 5: Synthesis of (R)-N-hydroxy-4-((3-(4-(4-(3-methylmorpholine))-6-morpholine-1,3,5-triazine-2-yl)phenyl)ureido)methyl)benzamide, the structural formula is as follows:

[0160]

[0161] Step 5 and operation are the same as Step 5 of Example 1, white solid, yield: 24%. NMR data are 1H NMR(400MHz,DMSO-d6)δ11.11(s,1H),9.14(s,1H),9.11(s,1H),8.77(s,1H),7.97-7.90(m,2H),7.75-7.64(m,4H),7 .32-7.25(m,2H),6.50(s,1H),4.78(s,1H),4.40(s,1H),4.32-4.27(m,2H),3.92(d,J=11.1Hz,2H),3.81(s,3H),3.7 2(d,J=11.4Hz,1H),3.66(t,J=5.6Hz,4H),3.58(d,J=11.8Hz,1H),3.44(d,J=11.7Hz,1H),3.19(t,J=12.8Hz,1H),1. 24(d,J=6.8Hz,3H).4.05(s,1H),3.98(s,1H),3.85(s,1H),3.83-3.74(m,9H),3.69(s,2H),3.55(s,1H),1.18(s,3H); 13 C NMR(100MHz,DMSO-d6)δ168.6,166.2,159.7,157.5,157.3,152.3,141.4,136.5,133.0,128.4,128 .1,128.0,125.1,120.1,74.0,70.7,66.7,66.5,46.2,43.7,41.8,38.6,14.5.HRMS(ESI)m / z:calcd for C 26 H 31 N8O5[M+H] + 549.2574; Found 549.2580.

[0162] The following experiments are used to illustrate the beneficial effects and applications of the compounds represented by the general formula (I) of the present invention.

[0163] Experimental Example 1 In vitro PI3Ks and mTOR kinase inhibition test:

[0164] The inhibitory activity of each kinase subtype (PI3Kα, β, γ and δ) of the compounds of the present invention was tested using the ADP-GloLuminescent Kinase Assay method. The specific experiments refer to the references (Eur. J. Med. Chem., 2020, 204, 112637; Eur. J. Med. Chem., 2022, 229, 114055). The reagents used in the kinase reaction are as follows: HEPES (50mM) pH 7.5 with NaCl (100mM), EGTA (1.0mM), MgCl2 (3.0mM), DTT (2.0mM) and CHAPS (0.03%). During the reaction, 50μM PIP2 and 25μM ATP were added to each 10mL of the test compound containing different concentrations (0.05nM-1.0μM). The reaction system was incubated at room temperature for 1h, and then 10μL of the reagent ADP-Glo ​​was added to terminate the enzyme reaction. Data were collected using Envision software, and GraphpadPrism 5 was used to analyze and fit the IC values ​​of the compounds. 50 value.

[0165] The Lance Ultra method was used to test the mTOR kinase inhibitory activity of the compound. The same reference method (Eur. J. Med. Chem., 2020, 204, 112637; Eur. J. Med. Chem., 2022, 229, 114055) was used. The reagents used in the kinase reaction were as follows: HEPES (50mM), NaCl (10mM), EGTA (1.0mM), MnCl2 (3.0mM), DTT (2.0mM) and Tween-20 (0.01%). Then, 50nMULight-4E-BP1 peptide and 8μM ATP were added to the compound test system of different concentrations (0.05nM-1.0μM) and diluted to a final volume of 10μL. The reaction system was incubated at room temperature for 45min, and then 10μL of detection leaves were added to terminate the enzyme reaction. Data collection and analysis were the same as the above PI3Ks kinase test.

[0166] The PI3Ks and mTOR kinase inhibitory activities of all examples are shown in Table 1. As can be seen from the table, most of the compounds have sodium molar level inhibitory activity against the above kinases, and are comparable to the positive controls PF-05212384 and PI-103. In particular, Example 2 has significantly better inhibitory activity against each kinase than the positive controls PF-05212384 and PI-103. This shows that the compounds of the examples of the present invention are highly effective pan-PI3K / mTOR inhibitors.

[0167] Table 1. PI3Ks and mTOR enzyme inhibitory activity of the example compounds (IC50, nM)

[0168]

[0169]

[0170] Experimental Example 2 In vitro HDAC enzyme inhibition activity test:

[0171] The HDACs enzyme inhibitory activity of the compounds of the present invention was evaluated by the Fluorescent-based HDACActivity Assay method, using SAHA, a broad-spectrum HDAC inhibitor on the market, as a positive control.

[0172] The test steps of enzyme inhibition activity are as follows: prepare DMSO solution of the compound to be tested, and prepare buffer solution, enzyme solution and corresponding substrate / trypsin mixed solution in sequence according to the instructions of the kit; add compound solution, enzyme solution and substrate / trypsin mixed solution of gradient concentrations into 384-well plate to prepare catalytic reaction system (with no compound control and no enzyme control wells); after incubation at room temperature for a certain period of time, use Synergy microplate reader to continuously read the fluorescence signal value, select the linear reaction segment to obtain the slope, and then calculate the inhibition rate at each concentration, IC 50 The values ​​were obtained by fitting with GraphPad Prism5 software.

[0173] The inhibitory activity of the compounds on different HDAC subtypes is shown in Table 2, from which it can be seen that the tested example compounds have submicromolar inhibitory activity on different HDAC subtypes. In particular, the tested example compounds have sodium molar inhibitory activity on the HDAC6 subtype, which is comparable to the positive control SAHA, and shows significant subtype selectivity.

[0174] Table 2. HDAC enzyme inhibitory activity (IC 50 , nM)

[0175] Example HDAC1 HDAC2 HDAC3 HDAC6 HDAC8 Example 1 730 1231 562 50 565 Example 2 763 831 530 28 364 Example 3 561 1013 720 45 461 Example 4 740 665 782 27 532 Example 5 465 813 604 36 395 SAHA 20 25 50 21 475

[0176] Experimental Example 3 Tumor cell line antiproliferative activity experiment:

[0177] The CCK-8 method was used to evaluate the inhibitory activity of the compounds on cell proliferation, and the half inhibitory concentration IC was determined by single-concentration activity screening and multiple-concentration determination. 50Value. The detection principle is as follows: Cytotoxicity (CCK-8 method) Detection principle: CCK-8 reagent contains WST-8, which is reduced to a highly water-soluble yellow formazan product (Formazan) by dehydrogenase in the cell mitochondria under the action of the electron carrier 1-methoxy-5-methylphenazine dimethyl sulfate (1-Methoxy PMS). The amount of formazan generated is proportional to the number of living cells.

[0178] The experimental method is as follows:

[0179] (1) Inoculation of cells: Prepare cells into a single cell suspension using culture medium containing 10% fetal bovine serum, inoculate 90 μL of 5×104 / mL adherent cells and 9×104 / mL suspension cells per well of a 96-well plate, and pre-culture at 5% CO2 and 37°C for 24 h.

[0180] (2) Add the sample solution to be tested: add 10 μL of sample solution to each well. Set one concentration for each sample in the initial activity screening, and set up 3 replicate wells; IC 50 Eight concentrations (including 0 concentration) were measured, and three replicate wells were set for each concentration; the cells were placed in an incubator and cultured for 48 hours. The experiment set up a blank group (Blank), a control group (Control), and a drug group (Drug).

[0181] (3) Color development: Aspirate the old culture medium and drug solution from adherent cells (add 10 μL CCK-8 stock solution directly to suspended cells), add 100 μL CCK-8 solution diluted ten times to each well, and continue culturing at 37°C, 5% CO2 for 1-4 h (keep away from light and observe in real time).

[0182] (4) Detection: Measure the absorbance at 450 nm using an enzyme-labeled instrument and record the raw data results.

[0183] (5) The original data were standardized using Excel software, and the cell proliferation inhibition rate was calculated by the OD value of each well in the initial screening (formula = (ODControl-ODDrug) / (ODControl-ODBlank)×100%), and the inhibition rate was statistically calculated. 50 The data were calculated by GraphPad Prism 8, and the experimental results are expressed as ± SD.

[0184] Table 3. Tumor cell line proliferation inhibition activity (IC 50 , μM)

[0185] Example Molm-13 K562 THP1 MKN-45 HCT116 Example 1 0.65 1.15 0.78 5.32 1.23 Example 2 0.81 1.20 0.90 2.51 3.35 Example 3 1.61 2.75 2.28 7.85 5.62 Example 4 1.31 2.30 8.75 4.52 3.20 Example 5 0.016 0.32 0.44 5.60 1.23 SAHA 1.50 2.10 5.40 6.30 12.10

[0186] Table 3 shows the antiproliferative activity of some of the example compounds on various tumor cell lines. It can be seen that the example compounds have micromolar level of proliferation inhibition activity on different tumor strains, especially high sensitivity to leukemia cell lines Molm-13, K562 and THP1. Among them, Example 5 showed sodium molar level inhibitory activity on the above three cell lines, which was significantly better than the positive control vorinostat (SAHA); Example 1 had submicromolar level inhibitory activity on tumor lines Molm-13 and THP1, which was significantly better than the positive control SAHA. It can be seen that the example compounds of the present invention can be potentially used in the clinical treatment of the above tumors.

[0187] In summary, the compounds protected by the present invention are multi-target inhibitors of PI3Ks / mTOR / HDACs, which can inhibit the proliferation of various solid tumors and blood tumor cells through multiple molecular mechanisms, have high sensitivity to blood tumors, especially acute myeloid leukemia, and can be potentially used in the clinical treatment of them.

[0188] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. For those skilled in the art, the present invention may have various modifications and variations. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included in the protection scope of the present invention.

Claims

1. A diaryl urea PI3K / mTOR / HDAC multi-target inhibitor, characterized in that: A structure containing one of the following: (1) R1 is R2 is X is n is 0; (2) R1 is R2 is X is n is 0; (3) R1 is R2 is X is n is 0; (4) R1 is R2 is X is n is 0; (5) R1 is R2 is X is n is 0.

2. The method for preparing the diaryl urea PI3K / mTOR / HDAC multi-target inhibitor according to claim 1, characterized in that: Using cyanuric chloride (a) as a raw material, two chlorine atoms of the raw material (a) react with morpholine or other nitrogen-containing heterocyclic rings through substitution reaction to introduce R1 group to obtain intermediate (b); on the basis of intermediate (b), R2 group is introduced through nucleophilic substitution reaction to obtain intermediate (c); intermediate (c) and 4-aminophenylboronic acid pinacol ester undergo Suzuki coupling reaction to obtain intermediate (d); the amino end of intermediate (d) is connected with 4-aminobenzoic acid methyl ester through urea group to obtain intermediate (e); intermediate (e) and hydroxylamine undergo substitution reaction to obtain target compound (f); the preparation route is as follows:

3. A pharmaceutical composition comprising at least one pharmaceutically acceptable excipient and an effective therapeutic dose of at least one diaryl urea PI3K / mTOR / HDAC multi-target inhibitor as claimed in claim 1.

4. Use of the diaryl urea PI3K / mTOR / HDAC multi-target inhibitor according to claim 1 or the pharmaceutical composition according to claim 3 in the preparation of a medicament for preventing and / or treating and / or adjuvant treating proliferative diseases, metabolic diseases, nervous system diseases and tuberous sclerosis caused by overactivation of mTOR kinase and HDAC.

5. The use according to claim 4, characterized in that The proliferative diseases are blood cancer cells, colon cancer cells, and gastric cancer cells.

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