WO2013170758A1

WO2013170758A1 - Fused ring-structured benzamide compound and application thereof as antineoplastic medicament

Info

Publication number: WO2013170758A1
Application number: PCT/CN2013/075688
Authority: WO
Inventors: 李建其; 张庆伟; 贾志丹; 周斌; 黄道伟; 王淑娟
Original assignee: 国药一心制药有限公司; 上海医药工业研究院
Priority date: 2012-05-18
Filing date: 2013-05-16
Publication date: 2013-11-21
Also published as: CN103420917A; CN103420917B

Abstract

Disclosed are a fused ring-structured benzamide compound and an application thereof. Pharmacological experiments show that the compound of the present invention provides great histone deacetylase 1 (HDAC1) inhibitory activity, weak inhibition against normal cells, reduced toxicity, should provide further reduced toxic side effects when applied as an antineoplastic medicament, and further facilitates use as the antineoplastic medicament. The present invention provides great efficacy in the treatment of diseases induced by abnormal gene expression, such as: tumors, endocrine disorders, immune system diseases, genetic diseases, and neurological diseases. The fused ring-structured benzamide compound is a compound as represented by formula (V) or a pharmaceutically acceptable salt thereof.

Description

Benzoamides containing fused ring structure and their application as antitumor drugs

The present invention relates to a method for synthesizing benzamides containing a fused ring structure and the use of such compounds for histone deacetylase inhibitory activity and for treating malignant tumors and diseases associated with differentiation and proliferation. Background technique

The causes of tumorigenesis and development are very complicated, and in the final analysis, they are closely related to gene expression and abnormal activity of gene expression products. The factors that lead to abnormal tumor gene expression and gene expression product activity come from two major changes, namely genetics and epigenetics. Genetic alteration refers to changes in gene sequences, such as gene mutations, deletions, and recombination, which play an important role in certain types of tumorigenesis. The results of recent studies have become increasingly clear that epigenetic changes, such as the effects of environmental pollution on the human body, have a more general significance in the occurrence and development of tumors, and the occurrence, development, prognosis and outcome of tumors are not only Depending on genetic factors, it is also affected by epigenetic modifications. Epigenetics refers to the regulation of a type of gene expression that affects the transcriptional activity of a gene without involving DNA sequence changes. Its molecular basis mainly involves two aspects: one is for methylation modification of DNA, and the other is for staining. Acetylation of histones. Histone acetylation and deacetylation of chromatin is one of the key links regulating gene expression, and two types of enzymes determine the degree of histone acetylation, namely Histone acetyltransferases (HAT) and sputum group. Protein deacetylase (Histone deacetylases, HDAC). Acetylation of histones activates the transcription of specific genes, while HDAC inhibits the transcriptional expression of genes. At the same time, HDAC also plays an important role in the acetylation-deacetylation process of non-histone proteins, including transcription factors, signaling proteins, DNA repair enzymes, etc., and these target proteins play a decisive role in the regulation of gene expression. In conclusion, HDAC plays an extremely important role in epigenetic regulation through the effects of histone and non-histone acetylation processes, and abnormalities in this regulatory mechanism are associated with tumors. The occurrence and development of the drug are closely related. The research and development of small molecule drugs based on HDAC, an important molecular target that affects epigenetics, has become a hot spot in the field of international cancer targeted therapy. HDAC is a large family of enzymes. Its members are currently known in four major classes of 18 different subtypes, of which class I includes HDAC 1, 2, 3, and 8 subtypes, and class II includes HDAC4, 5, 6, 7, 9 and 10 subtypes (4, 5, 7, 9 belong to IIa, 6, 10 belong to lib); Class IV only has HDAC 1 1 subtype, which has certain same with the first two Source; Class III includes seven subtypes of Sirtl -7, with no structural homology to the former class III. HDACs have many biological functions, and different subtypes of HDACs play different biological roles in cells. Among them, Class I and Class II are most closely related to the occurrence and development of tumors. Recent studies have shown that HDAC overexpression or activity abnormalities play an important role in the occurrence and development of leukemia and solid tumors, while inhibition of HDAC functional activity shows significant anti-tumor effects in vitro and in vivo. Clinically studied HDAC inhibitors can be classified into four categories according to their chemical structure, 艮P:

(1) Hydroxamic acids, such as trichostatin (TSA), SAHA;

(2) a cyclic tetrapeptide such as Apicidin;

(3) short-chain or aromatic fatty acids, such as sodium butyrate;

(4) benzamides, such as MS-275;

There are currently two HDAC inhibitors, the hydroxamic acid Vorinostat and the cyclic peptide Depsipeptide, which were approved by the US FDA in 2007 and 2009 for the application of cutaneous T lymphoma (CTCL) as an indication for the treatment of solid tumors. The application is also in clinical trials, which marks the end of the conceptual validation phase of HDAC as a novel drug target, and also indicates that HDAC inhibitors have broad development as antitumor drugs.

j? , . Due to the similarity of histone deacetylase subtype structures, most of the existing histone deacetylase inhibitors such as Vorinostat and Depsipeptide and other candidate compounds in clinical studies are often pan-inhibitors, usually inhibited simultaneously. Multiple subtypes, affecting swollen While tumor growth and metastasis, it often interferes with normal physiological functions and has potential toxic side effects. Therefore, the development of HDAC enzyme subtype inhibitors can not only reduce toxic side effects, but also provide probes for the biological function of each subtype, which has far-reaching significance. Studies have shown that benzamide HDAC inhibitors can selectively act on histone deacetylase subtypes, especially the tumor-associated subtype HDAC1, inhibit proliferation of various tumor cells, induce tumor cell differentiation and/or apoptosis. With low in vivo toxicity, its representative drug MS-275 has entered the lib phase of clinical trials. Therefore, the design and synthesis of highly selective histone deacetylase inhibitors, especially benzamide histone deacetylase inhibitors, to obtain a new anti-malignant agent with good efficacy, low toxicity and high safety It is currently a research hotspot and technical difficulty in this field. Scientific practice has found that due to the specificity of targeted drugs, it is not possible to directly obtain another series of compounds with corresponding activities or activities due to simple modification of a structure in a compound, and it is necessary to consider the difference between the structure of the compound and the spatial conformation of the target protein. And key binding sites, design, synthesis and biological screening to obtain a series of active new derivatives. In view of the important influence of the enzyme recognition region on the activity of the benzamide HDAC inhibitor structure, in the substrate structure bound to the enzyme recognition region, it is possible to obtain a series of new compounds with better activity if certain substituent changes are made. Furthermore, in view of the diversity of tumor cells and related cancers, compounds having the same enzyme inhibitory activity may have specific or selective antiproliferative effects on different tumor cells due to structural differences. Therefore, the design of compounds of structural diversity of benzamide HDAC inhibitors has important scientific value. A Chinese patent application (Application No. 201 110175233.1) filed by the inventor of the present invention discloses a class of benzamide histone deacetylase inhibitors, and it is found that the selectivity of HDAC enzyme needs further improvement, and according to clinical application. There is a need to design and synthesize more active, different structural types of benzamide histone deacetylase inhibitors to maintain or enhance the inhibitory activity of HDACs and HDAC1 enzymes. To find compounds that not only have better histone deacetylase subtype inhibitory activity, but also produce specific or selective antiproliferative effects on different tumor cells, and look forward to the development of an ideal antitumor drug. Summary of the invention

SUMMARY OF THE INVENTION The object of the present invention is to disclose a benzamide compound having a fused ring structure and its use as an antitumor drug to overcome the drawbacks of the prior art and to meet the needs of clinical applications. The benzoic acid compound containing a fused ring structure according to the present invention has the formula (V):

(V)

among them:

! ^ represents hydrogen, halogen, amino, hydroxy, nitro, cyano, fluorenyl of 1 to 4 carbon atoms, decyloxy of 1 to 4 carbon atoms, amino fluorenyl of 1 to 4 carbon atoms, 1 to a mercaptoamino group of 4 carbon atoms, an acyl group of 2 to 4 carbon atoms, an acylamino group of 2 to 4 carbon atoms, a thioindenyl group of 1 to 4 carbon atoms, a trifluoromethyl group, 1 to 4 carbons a carboxyl group of an atom, a decyloxycarbonyl group of 1 to 4 carbon atoms, a phenyl group or a heterocyclic ring;

Y or M is N or C;

X is O, S or N;

Xi is gas, halogen;

The term "heterocyclic ring" refers to a saturated or unsaturated heterocyclic ring containing one or more heteroatoms (nitrogen, oxygen, sulfur);

The halogen is preferably fluorine, chlorine, bromine or iodine;

The mercapto group of 1 to 4 carbon atoms, preferably methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl or tert-butyl;

The methoxy group of 1 to 4 carbon atoms is preferably a methoxy group, an ethoxy group or a n-propoxy group. Isopropoxy, n-butoxy or isobutoxy;

The aminoguanidino group of 1 to 4 carbon atoms is preferably an aminoethyl group, a 1-aminopropyl group or a 2-aminopropyl group;

The mercaptoamino group of 1 to 4 carbon atoms is preferably N-methylamino, N-ethylamino or N-isopropylamino;

The acyl group of 2 to 4 carbon atoms is preferably an acetyl group, a propionyl group or an isobutyryl group; and the acylamino group having 2 to 4 carbon atoms is preferably an acetylamino group, a propionylamino group, a butyrylamino group or an isobutyryl group. Amino group;

The thioindenyl group of 1 to 4 carbon atoms is preferably a methylthio group, an ethylthio group or a propylthio group;

The compound of the formula V can be salted with a mineral acid or an organic acid to obtain a salt form of a compound of the formula V, which is a hydrochloride, a hydrobromide, a sulfate, an acetate, Lactate, tartrate, citrate, citrate, trifluoroacetate, malate, maleate, succinate, p-toluenesulfonic acid or methanesulfonate. In order to facilitate the understanding of the present invention, the following specific compounds are preferred from the compounds of the formula V, but the benzoic acid compound containing a fused ring structure of the present invention is not limited to the following compounds:

V-l N-(2-aminophenyl)-4-((2-methyl-1H-benzimidazole-6-carboxamido)methyl)benzenecarboxamide,

V-2 N-(2-aminophenyl)-4-((1Η-benzimidazole-6-formylamino)methyl)benzamide,

V-3 N-(2-amino-5-fluorophenyl)-4-((2-methyl-1H-benzimidazole-6-carboxamido)methyl)benzamide,

V-4 N-(2-amino-5-fluorophenyl)-4-((2-(3-pyridyl)-1H-benzimidazole-6-formylamino)methyl)benzamide,

V-5 N-(2-aminophenyl)-4-((2-(3-pyridyl)-1H-benzimidazole-6-carboxamido)methyl)benzamide,

V-6 N-(2-aminophenyl)-4-((2-(isopropyl)-1H-benzimidazole-6-carboxamido)methyl)benzamide,

V-7 N-(2-Amino-5-fluorophenyl)-4-((1Η-benzimidazole-6-formylamino)methyl)benzamide, V-8 N-(2-Amino-5-fluorophenyl)-4-((2-(isopropyl)-1H-benzimidazole-6-carboxamido)methyl)benzamide,

V-9 N-(2-aminophenyl)-4-((2-(6-fluoro-3-pyridyl)-1H-benzimidazole-6-carboxamido)methyl)benzamide,

V-10 N-(2-Amino-5-fluorophenyl)-4-((2-(6-fluoro-3-hydroxypyridyl)-1H-benzimidazole-6-formylamino)methyl Benzoylamide,

V-l l N-(2-aminophenyl)-4-((2-(4-fluorophenyl)-1H-benzimidazole-6-carboxamido)methyl)benzamide,

V-12 N-(2-Amino-5-fluorophenyl)-4-((2-(4-fluorophenyl)-1H-benzimidazole-6-carboxamido)methyl)benzamide,

V-13 N-(2-aminophenyl)-4-((2-oxo-1,3-dihydro-1H-benzimidazole-6-formylamino)methyl)benzamide,

V-14 N-(2-Amino-5-fluorophenyl)-4-((2-oxo-1,3-dihydro-1H-benzimidazole-6-formylamino)methyl)benzene Amide,

V-15 N-(2-aminophenyl)-4-((2-(phenyl)-1H-benzimidazole-6-carboxamido)methyl)benzamide,

V-16 N-(2-Aminophenyl)-4-((1Η-吲哚-5-formylamino)methyl)benzamide, V-17 N-(2-amino-4-pyridyl) -4-((1Η-吲哚-5-formylamino)methyl)benzamide,

V-18 Ν-(2-amino-5-fluorophenyl)-4-((1Η-吲哚-5-formylamino)methyl)benzoamide,

V-19 Ν-(2-aminophenyl)-4-((1Η-benzofuran-5-formylamino)methyl)benzoylamine,

V-20 Ν-(2-amino-4-pyridyl)-4-((1Η-benzimidazole-6-formylamino)methyl)benzenecarboxamide,

Or a pharmaceutically acceptable salt of any of V-1 to V-20. The structural formula of the above compounds is shown in Table 1. Table 1 Preferred compound numbers and corresponding structural formulas

General Method 1: Synthesis of Intermediate III

Will replace acid I (50 mmol), O-(IH-benzotriazol-1-yl)-oxime, oxime, Ν', Ν'-tetramethylisourea hexafluoride (HBTU) (18.95g) , 50 mmol) was added to DMF, and triethylamine (10.1 g, 100 mmol) was slowly added dropwise under an ice bath, and stirred at room temperature for 1 hour to obtain a homogeneous reaction mixture. The aminomethylbenzoic acid II (50 mmol) was dissolved in 50 ml of a lmol/L aqueous solution of sodium hydroxide, and the above homogeneous reaction solution was added dropwise at the reaction temperature, and the mixture was stirred for 6 hours. The reaction solution was adjusted to pH 5-7 with hydrochloric acid to precipitate a large solid, which was filtered and dried to give Intermediate III. General Method 2: Preparation of Target Compound V

Intermediate 111 (1 mmol), compound IV (1 mmol), HBTU (0.379 g, 1 mmol) were sequentially added to 10 ml of N,N-dimethylformamide, and the mixture was kept in an ice bath and triethylamine was added dropwise.

(2 mmol), stirring was continued for another 4 hours at room temperature. The reaction solution was poured into 50 mL of water, and a solid was precipitated and filtered. The filter cake was washed successively with saturated sodium hydrogencarbonate, saturated brine and water for 1 to 2 times, and the residue was recrystallized from anhydrous ethanol to give the object compound V. The above preparation method may further comprise reacting a compound of the formula V with a mineral acid or an organic acid to cool a salt of the compound of the structure of the formula V. Ri, X, Y, Xi, and M in the above reaction method are as described above. Compounds I, II, IV and HBTU can be purchased commercially. Pharmacological tests have shown that the compounds of the present invention have a strong inhibitory effect on histone deacetylases (HDACs), especially histone deacetylase tumor-associated subtype HDAC1 (Example 21), some compounds such as V -2, V -3, V -8, V -10, V -12, V-16, V -17, V-18 and V-19 were superior to the positive control drug MS-275 in inhibiting HDACs and HDAC1. For example, compound V-16 has an inhibitory activity against HDAC1 of IC _5Q = 99 nM, which is significantly better than that of MS-275 (IC _5Q = 668 nM), indicating better histone deacetylase subtype targeting. Pharmacological tests showed that the compound of the present invention had strong differentiation and anti-proliferative activity against a plurality of tumor cells as compared with the positive control drug MS-275 (Example 22). Some compounds have better anti-tumor cell proliferation activity than the positive control drug MS-275, such as compounds.

V-2, V-3, V-8, V-10, V-16, V-19, V-20 have higher inhibitory activity against HCT116 human colon cancer cells, which is superior to MS-275. Among them, compounds V-2, V-3, V-10 and V-16 also showed superior antiproliferative activity against K562 human chronic myeloid leukemia cells and Jurkat E6-1 human T cell lymphoma. Pharmacological tests showed that the compound of the present invention had weaker inhibitory activity against normal cells than the control drug MS-275, and had lower toxic side effects (Example 23), indicating that the benzamide compound of the present invention was Tumor cells and normal cells have better selectivity in inhibiting proliferation, indicating that they have lower toxic side effects when used as antitumor drugs. Pharmacological tests showed that the compound of the present invention had a low acute toxicity in mice relative to the control drug MS-275 (Example 24). The LD50 of single-dose in Compound V-2, V-8, V-16, V-19 and V-20 mice was 2.63g/kg, 2.12g/kg, 1.95g/kg, 1.88g/kg, respectively. Mouth 2.14g/kg (reported in the literature MS-275 LD50=506 mg/kg, Current Medicinal Chemistry, 2003, 10(22), pp. 2343-2350). Pharmacological experiments show that the compounds of the invention have the following beneficial effects:

1) The compound of the present invention has good histone deacetylase (HDACs), especially histone deacetylase tumor-associated subtype HDAC1 inhibitory activity, and has good inhibitory activity against various tumor cells in human body.

2) Compared with the existing positive control drugs, the compounds of the present invention have a weak inhibitory effect on normal cells while inhibiting tumor cells, and exhibit better selective inhibitory activity, and have good anti-tumor clinical application. prospect.

3) Preliminary experiments on acute toxicity showed that the compound of the present invention has higher safety and less toxicity than the positive control drug MS-275. In summary, the compound of the present invention should have less toxic side effects and selectivity when used as an antitumor drug, and is more easily used as an antitumor drug. The tumor is a solid tumor and a hematoma, preferably colon cancer, liver cancer, lung cancer, breast cancer, esophageal cancer, gastric cancer, nasopharyngeal cancer, ovarian cancer, bladder cancer, rectal cancer, skin cancer, and lymphoma. The compounds of the present invention can be administered to mammals (including humans) in need of tumor treatment by oral, injection or the like in the form of a composition. The composition comprises a therapeutically effective amount of a compound of formula (V), or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier. The carrier refers to a carrier conventional in the pharmaceutical field, for example: a diluent, an excipient such as water, etc.; a binder such as a cellulose derivative, gelatin, polyvinylpyrrolidone or the like; a filler such as starch or the like; a cracking agent such as Calcium carbonate, sodium bicarbonate; in addition, other adjuvants such as flavoring agents and sweeteners may also be added to the composition. The composition of the present invention can be prepared into a conventional solid preparation such as a tablet, a capsule or the like for oral administration; it can also be prepared into an injection preparation or the like for injection. The various dosage forms of the compositions of the present invention can be prepared by conventional methods in the pharmaceutical art, wherein the compound of the active ingredient of the formula V is present in an amount of from 0.1% to 99.5% by weight based on the total weight of the composition. The compounds of the formula V according to the present invention can be administered to mammals (including humans) clinically by oral or injection means, particularly preferably orally. The dosage is 0.0001 mg/kg to 200 mg/kg body weight per day. The optimal dose will depend on the individual, usually at the beginning of the dose, and then gradually increase the amount. The invention is characterized in that the enzymatic surface recognition region of the benzamide histone deacetylase inhibitor is diversified to increase the spatial matching of the compound with the surface recognition region of the enzyme, especially to increase the binding ability to the enzyme subtype. After screening for HDAC1 enzyme subtype inhibition, it was found that this compound has strong inhibitory activity against HDAC1 enzyme, suggesting that the key amino acid binding residues in the surface recognition region of HDAC1 enzyme can be combined with the fused heterocyclic group such as -NH- , -O-, etc. for specific recognition and binding, indicating that the surface recognition region of the enzyme is replaced by a structure with different substituents on different heterocycles or heterocycles, and the benzamide histone deacetylation with stronger inhibitory activity of the enzyme subtype can be obtained. An enzyme inhibitor. In addition, the structural changes in the structure of the compound and the surface recognition region of the enzyme enable the novel structural compound to exhibit specificity and selectivity to tumor cells and normal cells while having enzyme inhibitory activity, thereby finding a good therapeutic effect and toxic side effects. Low, safe new anti-tumor drugs provide theoretical guidance and the feasibility of drug-making. An advantage of the present invention is that the compound and its medicinal preparation have a good therapeutic effect for treating diseases caused by abnormal gene expression, such as tumors, endocrine disorders, immune system diseases, genetic diseases and nervous system diseases. detailed description

The present invention will be further described in detail below with reference to the embodiments, but the embodiments of the invention are not limited thereto. i:/:/ O 889s/-i>l£ 8s/-0/J20iAV

赃 ^ ^3⁄4*3⁄4 ϋ 蝴蝴蝴蝴 ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ------ ------ ------ ------ ------ ------ ------ ------ ------ ------ ------ ------ (9ΐS inch Η -- - - --

Οΐ

o

Oo

Example 5

Synthesis of V-5 N-(2-aminophenyl)-4-((2-(3-pyridyl)-1H-benzimidazole-6-formylamino)methyl)benzamide

4-((2-(3-Pyridyl)-1H-benzimidazole-6-carboxamido)methyl)benzoic acid (0.372 g, 1 mmol), o-phenylenediamine (0.108 g, 1 mmol), HBTU (0.455g, 1.2 mmol), triethylamine

(0.404 g, 4 mmol) was used as a raw material, and V-5 was synthesized according to the general method, and the yield was 55.3%. 0.21 g of the above compound V-5 was dissolved in hot ethanol, and 4 mol/L of hydrobromic acid/ethyl acetate (2 ml) was added dropwise thereto, followed by cooling to precipitate a hydrobromide salt of V-5.

ESI-MS [M+H] ⁺ : m/z 463.30

1H NMR (400MHz, DMSO-d ₆ ) 5ppm:

4.58 (d, 2H, J = 8.0 Hz), 4.97 (s, 2H), 6.60 (t, lH,

J = 8.0 Hz), 6.78 (d, lH, J = 8.0 Hz), 6.97 (t, lH, J = 8.0 Hz), 7.17 (d, lH, J = 8.0 Hz), 7.48 (d, 2H, J = 8.0 Hz), 7.64 (m, 2H), 7.83 (m, 2H), 7.97 (d, 2H, J = 8.0 Hz), 8.14 (s, lH), 8.35 (d, lH, J = 8.0 Hz), 8.53 (t, lH, J = 4.0 Hz), 8.73 (t, lH, J = 4.0 Hz), 9.19 (m, lH), 9.65 (s, lH) Example 6

Synthesis of V-6 N-(2-aminophenyl)-4-((2-(isopropyl)-1H-benzimidazole-6-carboxamido)methyl)benzamide

4-((2-(isopropyl)-1H-benzimidazole-6-carboxamido)methyl)benzoic acid (0.337 g, 1 mmol) (prepared according to general procedure), o-phenylenediamine (0.108) g, l mmol), HBTU (0.455 g, 1.2 mmol), triethylamine (0.404 g, 4 mmol) were used as starting materials, and V-6 was synthesized according to the general method 2. The yield was 29.3%. 0.20 g of the above V-6 was dissolved in hot ethanol, 98% sulfuric acid (1 ml) was slowly added dropwise, and the sulfate of V-6 was precipitated by cooling.

ESI-MS [M+H] 十: m/z 428.2108 91

.激 ώ 3⁄4 8-Λ^3⁄4Ψ1⁄2咩^ 'uiraoe οε

°%Γ9

ⁱ 0V0)m

2 three-(lorara

ΠΙΗΗ I rara 1'§9 '0) two 3⁄4 ^ book, (iorara

Γ3⁄4εε·ο) ώ ⁄ 3⁄4 (requires ώ (requires 3⁄4 classes ώ 3⁄4 ^*-ΗΪ-(3⁄4 Η)Η ςζ

8 m^

(Hl's)Z.S'6'( ^z H0'8=r'Hl'l)^r6'(Hl'ra)i7£'8'(Hl's)^r8'( ^z H0'8

=Γ3⁄4'Ρ)ς6'Ζ.'(Ηΐ'^)6Ζ.·Ζ.'(ζΗ0=Γ'Ηΐ'Ρ)εΖ.'Ζ.'( ^ζ '0'8=Γ'Ηΐ'Ρ)09'Ζ.'( ^ζ Η0'8

Raddg ( ⁹ p-oswa ' noop) ring κ Η _Τ

niaH I rara Γ§9 ·0) two 3⁄4

Iorara ΐ'§60ε'0)^ώ3⁄4(3⁄4ώ(3⁄4ϋ3⁄4ώ-9-^3⁄4^3⁄4-Ηΐ)Η

L m^

(Hi's)

L9'6'(Hl's)60'6'(Hl's)lr8'( ^z H0'8=r'H^P)S6'Z.'( ^z H0'9I=r'Hrp)lZ.- (zH ς 0 ^=Γ'ΗΓΡ)Ζ^·/ ^ζ Η0·8=Γ'Ηε'Ρ)ΐ7ΐ7·ζ;(ζΗ0·8=Γ'ΗΓΡ) ·Ζ ^ζ Η0·8=Γ'ΗΓΐ)

86·9'( ^ζ Η0·8=Γ'ΗΓΡ)8Ζ/9'( ^ζ Η0·8=Γ'ΗΓΐ)09·9'(ΙΚ'δ)06ν( ^ζ Η0·8=Γ

'H^'P)9S '(Hl'ra)0^-£'( ^z H0'8=r'H9'l)S£'I

Raddg ( ⁹ p-oswa ' noop) 3⁄4WM H _T

889S .0/CT0ZN3/X3d 8SL0LI/£10Z OAV ESI-MS [M+H] 十: m/z 446.32

1H NMR (400MHz, DMSO-d ₆ ) 5ppm:

1.36 (t, 6H, J = 8.0 Hz), 3.19 (m, lH), 4.57 (d, 2H,

J = 8.0 Hz), 5.24 (s, 2H), 6.36 (t, lH, J = 8.0 Hz), 6.54 (dd, lH, J = 4.0, 12.0 Hz), 7.11 (t, lH, J = 8.0 Hz) , 7.45 (d, 2H, J = 8.0 Hz), 7.55 (d, lH, J = 8.0 Hz), 7.79 (m, 2H), 7.95 (d, 2H, J = 8.0 Hz), 8.14 (s, lH) , 9.12 (t, lH, J = 8.0 Hz), 9.61 (s, lH) Example 9

Synthesis of V-9 N-(2-Aminophenyl)-4-((2-(6-fluoro-3-pyridyl)-1H-benzimidazole-6-formylamino)methyl)benzamide

4-((2-(6-fluoro-3-pyridyl)-1H-benzimidazole-6-carboxamido)methyl)benzoic acid (0.390 g, 1 mmol) (prepared according to General Procedure) O-phenylenediamine (0.108 g, 1 mmol), HBTU (0.455 g, 1.2 mmol), triethylamine (0.404 g, 4 mmol) was used as a starting material, and V-9 was synthesized according to General Method 2, yield 43.0%.

ESI-MS [M+H] ⁺ : m/z 481.30

1H NMR (400MHz, DMSO-d ₆ ) 5ppm:

4.59 (d, 2H, J = 8.0 Hz), 4.91 (s, 2H), 6.54 (d, lH,

J = 12.0 Hz), 6.60 (t, lH, J = 8.0 Hz), 6.79 (d, lH, J = 8.0 Hz), 6.98 (t, lH, J = 8.0 Hz), 7.17 (d, lH, J = 8.0 Hz), 7.45 (d, 2H, J = 12.0 Hz), 7.55 (d, lH, J = 8.0 Hz), 7.65 (d, lH, J = 4.0 Hz), 7.80 (t, lH, J = 8.0 Hz) ), 7.96 (d, 2H, J = 8.0 Hz), 8.11 (s, lH), 8.20 (dd, lH, J = 4.0, 8.0 Hz), 8.27 (d, lH, J = 8.0 Hz), 9.15 (t , lH, J = 8.0 Hz), 9.66 (s, lH) Example 10

V-10 N-(2-Amino-5-fluorophenyl)-4-((2-(6-fluoro-3-pyridyl)-1H-benzimidazole-6-carboxamido)methyl)benzene Synthesis of formamide

4-((2-(6-fluoro-3-pyridyl)-1H-benzimidazole-6-carboxamido)methyl)benzoic acid (0.390 g, 1 mmol), 4-fluoro-phenylenediamine (0.126g, 1 mmol), HBTU (0.455g, 1.2mmol), triethylamine (0.404g, 4mmol) as raw material, V-10 according to the general method, yield 81

2 three-(lorara

ΠΙΗΗ , (I rara ι'§9 ·0) two 3⁄4 ^ book, (iorara

Ώ-9-^3⁄4 3⁄4-HI- (requires 3⁄4 books)-B))卞 (required) -B) -N -Λ

(ΗΙ'δ)ς9·6'(ΗΓδ) Γ6'( ^ζ Η0·8=Γ'Η^ 9 8'(ΗΙ'δ)

0·8=Γ'ΗΓΡ) ΓΖ ^Ζ Η0·8=Γ'ΗΙ'1) 6·9'( ^Ζ Η0·8=Γ'ΗΙ'Ρ)8Ζ/9'( ^Ζ Η0·8=Γ

'Hl'l)09'9'te's)^6 '( ^z H0 =f'H^'P)09

Raddg ( ⁹ p-oswa ' noop) ring κ Η _τ

^8Γ081 z/ra :+[H+W] SW—IS3

Π^ςςνθ) niaH I rara Γ§80Γ0) 2⁄4^, ^ Array 3⁄43⁄4®) (I ⁰

'(ζΗ0·8=Γ'Η 'Ρ)ς6·Ζ ^ζ Η0·8=Γ'ΗΓΐ)8Ζ/Ζ ^ζ Η0·8=Γ'ΗΓΡ)ς9·/ ^ζ Η0·8=Γ'ΗΙ

'Ρ)ΐς·Ζ.'(ζΗ0'8=Γ'Η^Ρ)9ΐ·Ζ.'( ^ζ Η0'8=Γ'ΗΐΌΐΐ'Ζ.'(Η^)ες·9'( ^ζ Η0'8 =Γ

'Hl'l)9£'9'te's)6^'S'(zH0 =r'H^'P)6S ξ raddg ( ⁹ p-oswa ' noo ) ring κ Η _τ

968ΓΖ.617 z/ra H+W] S —IS3

°%0'0S

889S.0/CT0ZN3/X3d 8S.0.l/CT0Z OAV 61

°% ££ οε

, (Iorara

Π^ςςνθ) niaH, (Iorara Γ§9 ·0) 2⁄4 ^ book, (iorara ΐ'§Π£'θ) w^ mm ώ *(3⁄4 ώ ( n

( ^ζ ΐ0 ΐ=Γ'Η^Ρ)06Όΐ'(Ηΐ'δ)ΐ79'6'( ^ζ Η0'8=Γ'ΗΐΊ)

Ϊ́ΐ7·Ζ/(ζΗ0·8=Γ'ΗΓΡ)9ΓΖ (Η^ι 86·9'( ^ζ Η0·8=Γ'ΗΓΡ)8Ζ;9'( ^ζ Η0·8=Γ

'Hl'l)09'9'te's)06 '( ^z H0 =r'H^'P)£S

: raddg ( ⁹ P"OSWa 'ZH 001) Ring K Η _τ

8ΖΎ0Ρ z/ra: +[H+W] Sn-ISH

. %ε·% ^ji 'ει-Λ ζ\

ΠΙΗΗ , (Iorara Γ§80Γ0) 2⁄4 , (^ 阵 3 3⁄43⁄43⁄4) (I ^orara ΐ'§Π£'θ)

(Hl's)8S'6'(Hl'ra)sr6'(zH0'8=r'H^l)^'8'(Hl's)0r8'( ^z H0'8

=Γ'Η 'Ρ'96)/ ^ζ Η0·8=Γ'ΗΓΐ)ε8·Ζ ^ζ Η0·8=Γ'ΗΓΡ)εΖ/Ζ ^ζ Η0·8=Γ'ΗΓΡ)09·

'Hl'l)9£'9'te's)1⁄2-S'(zH0 =r'H^'P)6S

: raddg ( ⁹ P"OSWa 'ZH 001) Ring K Η _τ

889S.0/CT0ZN3/X3d 8S.0.l/CT0Z OAV

+/∞3ζ inch [sSHI3+i : z - one gOs inch) wa circle H :idd- ΡΡΡΓ)ΠΛ0∞Γ)669(0ΤΓ0Γ)19Χ0∞ί3⁄4ΖΗί3⁄4Ηί3⁄4Ηί SΪΖΖΖ ====....

Pp(0∞r)∞(r)0(0∞(0∞i3⁄4z/HfHsz/Hz; Hi3⁄4z/HsZZZ ===......

p (0)060r(r)6X0∞n)s6X0∞Hi3⁄4Hz^Hi3⁄4HsZZZ==......

赃无^3⁄4*3⁄4 w ϋ 蝴蝴蝴 ΐ ΐ ΐ ΐ ΐ ΐ ΐ ΐ ΐ ΐ ΐ ΐ ΐ ΐ ΐ ΐ ΐ ΐ ΐ ΐ ΐ ΐ ΐ ΐ ΐ ΐ ΐ ΐ ΐ ΐ ΐ ΐ ΐ ΐ ΐ ΐ ΐ ΐ ΐ ΐ ΐ ΐ ΐ ΐ ΐ ΐ ΐ ((9ΐ ()s inch Η Η ώ ------.

3⁄4^显3⁄4 ΐίl Γΐ inch 0) Γ∞0Γ0) xlςς maHΗ-οιπιπ§-otrnu ,.

A gOs inch) wa circle H: idd- pr) 099X6 inch XO inch) 09 inch HHHiHlsZ =.... pp(£K6(0∞r)£∞(0r)99((YHz/HrHz; HrHz/Hz/ HSZZZ==.....

o m

赃不 * * ϋ 蝴蝴蝴 ΐ ( (ΐ)) inch (ά 9ΐ ΗΜ---,

3⁄43⁄4 feed 赃 i i butterfly 3⁄4^ ώ ώϋf inch 6Γ0) ( (ΐ)) inch Η ----

+/ 9∞£[sSHI3+111 : N - a gp (Os inch) wa circle H :idd9-

Ppn)£9X)9I(9) HsHHsi3⁄4z^sZ=.-. r)0r∞0∞HHi3⁄4zz.s^Z= ...

ΐΐH Bu

o o m

*城ϋ蝴蝶Λΐ)) inch (ά∞ΐ ΗςΜ-- ---- ώ ώϋ ((ΐ)) inch Η--- Zz

: raddg ( ⁹ p-oswa 'mnoo ) 3⁄4WM H _T

Ϊ́ΐ9ΐ'98£ z/ra: +[H+W] sw-isa ^ OVO)MV≡ ^iorara Z'l^iiVO) ΠΙΗΗ , (I.rara Γ§60Γ0)3⁄4 Required) II. (Iorara ΐ'§ 60ε'0)^ώ3⁄4(3⁄4ώ(3⁄4ϋ3⁄4ώ-9-^3⁄4^3⁄4-Ηΐ)Η

(Hi's)

^{9ΓΠ '(ζ Η0 · 9 =} Γ'ΗΓΐ) 06 · 8'(ΗΙ'δ) 6Γ8 '(ζ Ηΐ7 · 8 = Γ'Η' Ρ) ε6 · Ζ ζ Ηΐ7 · 8 'ζ Η9 · Ι = Γ' Η 'ΡΡ)89·Ζ (Ηΐ'ΐΐι)εΐ7·ζ;(ζΗ9·=Γ'ΗΓΡ) ΓΖ ζΗΐ·8' ^ζ Η Ι=Γ'ΗΓΐΡ) 6·9

Hl'P)lS-9'(H^s)£8 '( ^z H9'S=r'H^P)lS : raddg ( ⁹ P"OSWa 'ZH 001) Ring K Η _τ

9P9VS8E z/ra: +[H+W] SW-IS3

°%0·" ^ι '6Ι-Λ ^ : 'U^ (l ^omm ^ OVO three ^(lorara ΖΊ^ςςνθ) ΠΙΗΗ , (Iorara I'ggorO) two 3⁄4^ ' (^ array one

(HI's) 9 n

'(HI 's)89'6'( ^z H0'9=r'HI'l 6·8'(Η I's)6 Γ8'(Ηΐ'δ)θ Γ8'( ^Ζ Η0'8=Γ'Η^ 'Ρ)

'(HI's)W9'(H s rS'(zH9'S=r'H P)

: raddg ( ⁹ p-oswa ' noop) 3⁄4WM H _T

889S .0/CT0ZN3/X3d 8SL0LI/£10Z OAV

o

As can be seen from the above Table 2, the compounds of the present invention which were randomly selected for testing have strong inhibitory activities against HDACs and HDAC1, and the inhibitory activity against HDAC1 is remarkable, and some compounds such as V-2, V-3, V-8 V-10, V-12, V-16, V-17, V-18 and V-19 were superior to the positive control drug MS-275 in inhibiting HDACs and HDAC1. Example 22

In vitro inhibitory activity test of compound tumor cells

Determination of some compounds against PC-3 human prostate cancer cells, A549 human lung cancer cells, Colo320 human rectal cancer cells, Hep3B2.1-7 human liver cancer cells, HCT116 human colon cancer cells, K562 human chronic myeloid leukemia cells, Jurkat E6-1 Inhibition of human T cell lymphoma. The IC ₅ o value was measured by the CCK-8 method (Cat# CK04-13, Dojindo). The specific results are as follows: Table 3. Inhibition of in vitro proliferation of compounds on tumor cell lines IC _5Q assay

IC ₅₀ (μΜ)

Compound

PC-3 A549 Colo320 He 3B HCT116 K562 Jurkat E6-1

MS-276 0.6020 2.278 0.6222 6.816 0.6281 4.662 0.6614

V-2 0.242 2.626 2.460 2.461 0.0194 1.212 0.179

V-3 0.270 1.248 2.122 2.068 0.0888 2.092 0.128

V-8 2.226 21.22 3.896 16.89 2.462 17.11 2.122

V-10 10.896 2.126 1.089 1.622 0.2667 0.4269 0.4678

V-12 2.620 0.6244 9.242 2.760 2.778 2.884 1.269

V-16 0.1763 1.203 6.02 2.262 0.090 1.080 0.897

V-17 0.1914 1.109 5.987 10.22 1.188 1.908 0.998

V-18 0.969 0.2962 3.697 8.956 0.877 3.568 9.685

V-19 12.17 29.92 88.406 62.172 0.2211 9.061 2.262

V-20 2.668 6.089 26.997 16.997 0.0668 2.811 1.292 As can be seen from Table 3 above, some of the compounds of the present invention were tested before PC-3 Seven tumor cells, such as adenocarcinoma, have better anti-proliferative activity. Some compounds have superior anti-tumor cell proliferation activity than the positive control drug MS-275, such as compounds V-2, V-3, V-8, V-10, V-16, V-19, V-20 to HCT116 human colon cancer. The cells have higher inhibitory activity and are superior to MS-275. Among them, compounds V-2, V-3, V-10 and V-16 also showed superior antiproliferative activity against K562 human chronic myeloid leukemia cells and Jurkat E6-1 human T cell lymphoma. Example 23

Determination of the normal cell in vitro inhibitory activity of the compound:

The activity of the compound of the present invention against MCF10A (human normal mammary epithelial cell line) was determined.

The IC ₅₀ value was measured by the CCK-8 method (Cat# CK04-13, Dojindo). MS-275 was selected as the control drug for the in vitro inhibitory activity IC _5Q test of normal cell lines. The specific results are as follows (unit: μΜ): Table 4 In vitro inhibitory activity of the compound of the present invention and a control drug on normal cells

Num MCF10A

V-1 10.4

V-4 9.8

V-3 6.9

V-4 4.5

V-5 5.8

V-6 7.9

V-7 4.4

V-8 9.7

V-9 4.3

V-10 8.5

V-ll 6.6

V-14 4.5

V-13 7.7

V-14 6.9

V-15 8.4

V-16 5.4

V-17 6.4

V-18 7.7

V-19 18.3

MS-275 1.25 As can be seen from the above Table 4, the compounds V-1 to V-19 of the present invention tested were less active against normal cells than the control drug MS-275, and had lower toxic side effects, indicating the benzene of the present invention. Formamide compounds have better selectivity for inhibiting proliferation of tumor cells and normal cells, indicating that they have lower toxic side effects when used as antitumor drugs, and are easy to be used as tumor drugs. Example 24

Acute toxicity test: using the method reported by Zhang Juntian (Modern Pharmacological Experimental Method) (Beijing Medical University, China Union Medical University Joint Press, published in 1998), preliminary screening, using Bliss method statistics ("Practical pharmaceutical preparations" Technology, People's Health Publishing House, published in 1999), the LD _{50 of} single-dose in compound V-2, V-8, V-16, V-19 and V-20 mice was 2.63g/kg, 2.12, respectively. g/kg, 1.95 g/kg, 1.88 g/kg and 2.14 g/kg. Example 25

Tablet: Any compound selected from V-1 to V-20 lOOmg sucrose 150mg

Corn Starch 38mg

Calcium stearate 2mg

Preparation method: The active ingredient V-1 to V-20 is mixed with sucrose and corn starch, moistened with water, stirred uniformly, dried, pulverized and sieved, added with calcium stearate, uniformly mixed, and compressed. Each tablet weighs 290 mg and has an active ingredient content of 100 mg. Example 26

Injection: 15mg of any compound selected from V-1 to V-20 80mg of water for injection Preparation method: Dissolve any compound of active ingredient V-1 to V-20 in water for injection, mix well, filter, and obtain the solution Dispense under sterile conditions in ampoules, 95 mg per bottle, with an active ingredient content of 15 mg/bottle. The above embodiments are preferred embodiments of the present invention, but the embodiments of the present invention are not All changes, modifications, substitutions, combinations, and simplifications that are made without departing from the spirit and scope of the present invention are to be construed as equivalents, and are included in the scope of the present invention. Inside.

Claims

Claim

A benzamide compound having a fused ring structure, which is characterized by having a compound represented by the formula (V) or a pharmaceutically acceptable salt thereof:

(V)

among them:

Y or M is N or C; X is O, S or N; hydrogen or halogen.

The styrene-containing compound having a fused ring structure according to claim 1, wherein the halogen is fluorine, chlorine, bromine or iodine.

The benzoic acid compound containing a fused ring structure according to claim 1, wherein the fluorenyl group having 1 to 4 carbon atoms is a methyl group, an ethyl group, a n-propyl group or an isopropyl group. , n-butyl, isobutyl or tert-butyl.

The benzoic acid compound containing a fused ring structure according to claim 1, wherein the methoxy group of 1 to 4 carbon atoms is a methoxy group, an ethoxy group or a n-propoxy group. , isopropoxy, n-butoxy or isobutoxy.

The benzoic acid compound containing a fused ring structure according to claim 1, wherein the aminoguanidino group having 1 to 4 carbon atoms is an aminoethyl group, a 1-aminopropyl group or a 2- Aminopropyl.

The benzoic acid compound containing a fused ring structure according to claim 1, wherein the mercaptoamino group having 1 to 4 carbon atoms is N-methylamino, N-ethylamino or N- Isopropylamino.

The benzoic acid compound containing a fused ring structure according to claim 1, wherein the acyl group having 2 to 4 carbon atoms is an acetyl group, a propionyl group or an isobutyryl group.

The benzoic acid compound containing a fused ring structure according to claim 1, wherein the acylamino group having 2 to 4 carbon atoms is an acetylamino group, a propionylamino group, a butyrylamino group or an isobutyl group. Amido group.

The benzoic acid compound containing a fused ring structure according to claim 1, wherein the thioindenyl group having 1 to 4 carbon atoms is a methylthio group, an ethylthio group or a propylthio group. .

10. The benzoic acid compound having a fused ring structure according to claim 1, wherein the halogen is fluorine, chlorine, bromine or iodine;

The mercapto group of 1 to 4 carbon atoms is methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl or tert-butyl;

The methoxy group of 1 to 4 carbon atoms is a methoxy group, an ethoxy group, a n-propoxy group, an isopropoxy group, a n-butoxy group or an isobutoxy group;

The aminoguanidino group of 1 to 4 carbon atoms is an aminoethyl group, a 1-aminopropyl group or a 2-aminopropyl group;

The mercaptoamino group of 1 to 4 carbon atoms is N-methylamino, N-ethylamino or N-isopropylamino;

The acyl group of 2 to 4 carbon atoms is acetyl, propionyl or isobutyryl; the acylamino group of 2 to 4 carbon atoms is acetylamino, propionylamino, butyrylamino or isobutyrylamino ;

The thioindenyl group of 1 to 4 carbon atoms is a methylthio group, an ethylthio group or a propylthio group.

11. A benzamide compound having a fused ring structure, characterized in that it is selected from the group consisting of

V-2 N-(2-aminophenyl)-4-((1Η-benzimidazole-6-formylamino)methyl)benzamide,

V-7 N-(2-amino-5-fluorophenyl)-4-((1Η-benzimidazole-6-formylamino)methyl)benzamide,

V-8 N-(2-amino-5-fluorophenyl)-4-((2-(isopropyl)-1H-benzimidazole-6-carboxamido)methyl)benzamide,

V-15 N-(2-Aminophenyl)-4-((2-(phenyl)-1H-benzimidazole-6-carboxamido)methyl) Benzoylamide,

V-19 N-(2-Aminophenyl)-4-((1Η-benzofuran-5-formylamino)methyl)benzamide or

V-20 N-(2-Amino-4-pyridyl)-4-((1Η-benzimidazole-6-carboxamido)methyl)benzenecarboxamide.

Or a pharmaceutically acceptable salt of any of V-1 to V-20.

The benzamide compound having a fused ring structure according to any one of claims 1 to 11, wherein the salt is a hydrochloride, a hydrobromide, a sulfate or an acetate. , lactate, tartrate, citrate, citrate, trifluoroacetate, malate, maleate, succinate, p-toluenesulfonic acid or methanesulfonate.

The use of a benzoic acid compound containing a fused ring structure according to any one of claims 1 to 12 for the preparation of a medicament for treating tumors.

The application according to claim 13, wherein the tumor is a solid tumor and a hematoma, preferably colon cancer, liver cancer, lung cancer, breast cancer, esophageal cancer, gastric cancer, nasopharyngeal cancer, ovarian cancer, bladder cancer. , rectal cancer, skin cancer or lymphoma. A composition comprising a therapeutically effective amount of the benzoic acid compound having a fused ring structure according to any one of claims 1 to 12, and a pharmaceutically acceptable carrier.