WO2013170757A1

WO2013170757A1 - 4-aminoquinazoline hydroxamic acid compound and application as antineoplastic medicament

Info

Publication number: WO2013170757A1
Application number: PCT/CN2013/075687
Authority: WO
Inventors: 李建其; 张庆伟; 贾志丹; 周斌; 崔宁; 梁晓东
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: CN103420923B; CN103420923A

Abstract

Provided is a 4-aminoquinazoline hydroxamic acid compound as represented by formula (V) or a pharmaceutically acceptable salt thereof. The compound provides great histone deacetylase inhibitory activity, great inhibitory activities against various tumor cells of the human body, weak inhibition against normal cells, and reduced toxicity, and is applicable for development as an antineoplastic medicament. In addition, the compound also provides great efficacy in the treatment of diseases induced by abnormal gene expression.

Description

4-aminoquinazoline hydroxamic acid compound and application as antitumor drug

The present invention relates to a class of 4-aminoquinazoline hydroxamic acid compounds, and the use of such compounds as antitumor drugs. Background technique

Tumors are major diseases that threaten human health, and the treatment of tumors has been closely watched by the whole world. Traditional chemotherapeutic drugs non-specifically block cell division and cause cell death. While killing tumor cells, they also destroy normal human cells. And many cytotoxic drugs have a limited range of treatments, which can easily lead to treatment-related adverse reactions. In recent years, with the continuous improvement of molecular biology technology and the further understanding of the pathogenesis of tumors from the cellular and molecular levels, as well as the rapid development of technologies such as combinatorial chemistry, structure-based drug design and computer science, tumor biotherapy has Great progress has entered the era of molecular targeted therapy. Targeted anticancer drugs can target specific pathways, prevent tumor growth and reduce toxicity to normal cells. Their common features are: non-cytotoxicity and tumor cell targeting; cell regulation and stabilization; dose-limiting toxicity and maximum tolerated dose are generally not achieved in clinical studies; can kill chemotherapy insensitive or resistant Tumor cells have better effects in combination with conventional treatment (radiotherapy, chemotherapy). At present, many anti-tumor targets have been discovered, and histone deacetylase is an important target for the development of new anticancer drugs. The factors that lead to abnormal tumor gene expression and gene expression product activity come from two major changes, namely genetics and epigenetics. Among them, epigenetics refers to the regulation of a type of gene expression that affects the transcriptional activity of genes and does not involve changes in DNA sequences. The molecular basis is mainly related to two aspects: one is for methylation modification of DNA, and the other is Acetylation of chromatin histones. Chromatin group Protein acetylation and deacetylation are one of the key links regulating gene expression, and two types of enzymes determine the degree of acetylation of histones, namely Histone acetyltransferases (HAT) and histones. 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 influence of histone and non-histone acetylation processes, and the abnormality of this regulatory mechanism is closely related to the occurrence and development of tumors. The development of small molecule drugs, such as HDAC, which is an important molecular target that affects epigenetics, has become a hot spot in the field of international cancer targeted therapy. Histone deacetylase inhibitors can be classified into four classes according to their structure: benzamides, hydroxamic acids, fatty acids and cyclic peptides. SAHA (also known as Vorinostat) is a hydroxamic acid and is the first listed histone deacetylase inhibitor for the treatment of cutaneous T-cell lymphoma. Its application in the treatment of solid tumors is also in the clinical trial stage, which marks the end of the proof-of-concept research phase of HDAC as a novel drug target, and also indicates that HDAC inhibitors have broad development prospects as anti-tumor drugs. Hydroxamic acid HDAC inhibitors are composed of an aromatic ring, a fatty chain and a hydroxamic acid, which are an enzyme surface recognition region, a linking region and a metal binding region (zinc ion binding region). In order to find compounds that are more active than SAHA, the researchers have done a lot of research on this type of compound, in which the hydroxamic acid groups in the metal-binding region are kept unchanged, and the surface recognition and connection regions of the enzyme are optimized. In order to find more active, selective and safe derivatives, the main research direction of hydroxamic acid inhibitors. While increasing the anti-tumor activity, reducing the impact on normal tissues or cells is a subject of great concern. The invention aims to obtain a series of compounds by means of drug design and synthesis, and carry out compound inhibition and anti-tumor test in vitro to find that it is more developmental than the marketed drug SAHA. Think of anti-tumor drugs. Summary of the invention

It is an object of the present invention to disclose a 4-aminoquinazoline hydroxamic acid compound and its use as an antitumor drug for clinical applications. The 4-aminoquinazoline hydroxamic acid compound of the present invention has a compound as shown in (V)

Wherein, R ₂ , R ₃ , R ₄ or R ₅ is hydrogen, halogen, amino, nitro, hydroxyamino, —C ₄ decyloxycarbonyl, —aminodecyl, —indenyl, —C ₄ Acylamino, decyloxy, decyl, ureido, trifluoromethyl, d-C ₄ sulfonyl, arylsulfonyl, substituted phenyl, phenyl or heterocyclic;

n= 0~5 integer;

The substituted phenyl group has 1 to 4 substituents on the benzene ring, and the substituent is halogen, hydroxy, nitro, cyano, decyloxy, fluorenyl or amino group of 1 to 4 carbon atoms;

The heterocyclic ring refers to a saturated or unsaturated five-membered heterocyclic or six-membered heterocyclic ring containing one or more hetero atoms; the hetero atom is nitrogen, oxygen or sulfur;

The halogen is fluorine, chlorine, bromine or iodine;

The fluorenyl group refers to a branched, unbranched, and cyclic saturated hydrocarbon chain containing a specified number of carbon atoms, and the fluorenyl group of -C ₄ is preferably a methyl group, an ethyl group, a propyl group, an isopropyl group, or a ring. Propyl, butyl, isobutyl, sec-butyl, tert-butyl or cyclobutyl;

The oximeoxy group is a -C ₄ decyloxy group, a substituted benzyloxy group, a pyrrolidin-1-yl-(C ₂ -C ₄ )decyloxy group, a morpholin-1-yl-(C ₂ -C ₄ ) anthraceneoxy, piperazin-1-yl-(C ₂ -C ₄ )decyloxy, N-methylpiperazin-1-yl-(C ₂ -C ₄ )decyloxy or piperidine-1 -yl-(C ₂ -C ₄ )decyloxy;

The amino group means -NH _{2 ;} The -C ₄ aminoguanidino group is preferably an aminoethyl group, a 1-aminopropyl group or a 2-aminopropyl group; the -C ₄ decylamino group is preferably N-methylamino, N-ethylamino or N-isopropyl an amino group; amide group refers to a -C (O) NH -, " CC 4 amide group" as "C _r C ₄ alkyl with a" connected "amido", preferably acetylamino (C¾C (O) NH -), propionylamino (CH ₃ CH ₂ C(O)NH -), butyrylamino or isobutyrylamino;

The sulfonyl group means -SO ₂ - _; "sulfonyl group of -C4" means "mercapto group of -C ₄ " is bonded to "sulfonyl group", preferably methylsulfonyl group, ethylsulfonyl group. , propylsulfonyl, isopropylsulfonyl, n-butanesulfonyl, isobutylsulfonyl, sec-butylsulfonyl;

The salt is a hydrochloride, a hydrobromide, a sulfate, an acetate, a lactate, a tartrate, a citrate, a citrate, a trifluoroacetate, a malate, a maleic acid. Salt, 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 present invention is not limited to the following compounds:

V-l N-hydroxy-7-(quinazolin-4-amino)heptanamide,

V-2 N-hydroxy-7-(6,7-dimethoxyethoxyquinazolin-4-amino)heptanamide,

V-3 N-hydroxy-7-(6,7-dimethoxyquinazolin-4-amino)heptanamide,

V-4 N-hydroxy-6-(quinazolin-4-amino)hexanamide,

V-5 N-hydroxy-6-(6,7-dimethoxyquinazolin-4-amino)hexanamide,

V-6 N-hydroxy-7-(6-methylquinazolin-4-amino)heptanamide,

V-7 N-hydroxy-7-(6-chloroquinazolin-4-amino)heptanamide,

V-8 N-hydroxy-7-(6-fluoroquinazolin-4-amino)heptanamide,

V-9 N-hydroxy-7-(7-chloroquinazolin-4-amino)heptanamide,

V-10 N-hydroxy-7-(6-nitroquinazolin-4-amino)heptanamide,

V-l l N-hydroxy-7-(6-aminoquinazolin-4-amino)heptanamide,

V-12 N-hydroxy-7-(6-cyanoquinazolin-4-amino)heptanamide,

V-13 N-hydroxy-7-(6-acetamidoquinazolin-4-amino)heptanamide,

V-14 N-hydroxy-7-(8-chloroquinazolin-4-amino)heptanamide,

V-15 N-hydroxy-7-(5-fluoroquinazolin-4-amino)heptanamide or

V-16 N-hydroxy-7-(2-chloroquinazolin-4-amino)heptanamide.

The compound of the formula (V) may 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, or an acetic acid. Salt, lactate, tartrate, citrate, citrate, trifluoroacetate, malate, maleate, succinate, p-toluenesulfonic acid or methanesulfonate.

General method for synthesis of class V compounds:

Substituting 4-chloroquinazoline a (2.5 mmol), b (2.5 mmol), and 2 mL of triethylamine were added to isopropanol for 90 h at 90 °C. Evaporate the solvent under reduced pressure, and extract the solid with ethyl acetate/water. Set the organic layer. The solvent was removed to give an oil which was directly charged without purification. 10 mL of methanol was added to the oil and stirred in an ice-bath, and then KOH (0.42 g, 7.5 mmol), hydroxylamine hydrochloride (0.35 g, 7.5 mmol) and 1 mL of water were added. The reaction was carried out for 3 h in an ice bath, and the reaction mixture was adjusted to pH 4-5 with hydrochloric acid. The solid is recrystallized from acetonitrile or ethyl acetate/methanol (2: 1-5: 1) to give the title compound. The above preparation method may further comprise reacting a compound of the formula V with a mineral acid (or an inorganic base), an organic acid (or an organic base), and cooling the salt of the compound of the structure of the formula V. Ri, R ₂ , R ₃ , R ₄ , R ₅ , n in the above reaction method are the same as described above;

In the above preparation method, the raw materials involved, compounds a, b and hydroxylamine hydrochloride can be purchased through commercial channels;

Pharmacological tests have shown that the compounds of the present invention have a strong inhibitory effect on histone deacetylase (HD ACs) (Example 17), wherein compounds V-3, V-6, V-7, V-9 The inhibitory activity of V-10 and V-13 on HDACs was better than that of the positive control drug SAHA. Pharmacological tests have shown that the compounds of the present invention have strong differentiation and anti-proliferative activity against a plurality of tumor cells (Example 18). Some compounds are inhibiting Jurkat E6-1 (human T cell lymphoma), Hut78 (T lymphocytic leukemia cells), Colo320 (human rectal cancer cell line), Caki-1 (human renal cell carcinoma cells) relative to the positive control drug SAHA. Strain)

MDA-MB-435 S (human breast cancer cells), A549 (human lung cancer cells), PANC-1 (human pancreatic cancer cells) and the like have better activity in tumor cell proliferation, indicating the 4-aminoquinazoline isoform of the present invention Hydroxamic acids have good inhibitory activity against a variety of tumor cells. Pharmacological tests showed that the compound of the present invention has weak inhibitory activity against normal cells and has lower toxic side effects (Example 19), indicating that the 4-aminoquinazoline hydroxamic acid compound of the present invention and a positive control Compared with the drug SAHA, it has better selectivity for inhibiting proliferation of tumor cells and normal cells, indicating that it has lower toxic side effects when used as an antitumor drug. Pharmacological tests show that the compounds of the present invention have low acute toxicity in mice (Examples)

20). Compound V-2, V-7, V-9, V-16 mice were given LD ₅ in a single dose. They were 1.6 g/kg, l. lg/kg, 1.9 g/kg and 1.3 g/kg, respectively. Pharmacological experiments show that the compounds of the invention have the following beneficial effects:

1) The compounds of the present invention have good histone deacetylase (HDACs) inhibitory activity and have excellent inhibitory activity against various tumor cells in humans.

2) Compared with the hydroxamic acid market-based drug SAHA, the compound of the present invention has a weak inhibitory effect on normal cells while inhibiting tumor cells, exhibits good selective inhibitory activity, and has excellent antitumor activity. Prospects for clinical application.

3) Acute toxicity preliminary experiments show that the compounds of the present invention have higher safety and less toxicity in mice. 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 the 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 arts. The content of the compound having the active ingredient of the formula V is 0.1% by weight of the composition~

99.5% by weight. The compounds of the formula V according to the present invention can be administered clinically to mammals (including humans) 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 present invention combines the structural characteristics of SAHA, adopts amide bond exchange, and skeleton migration to transform the mother nucleus, and introduces an antitumor drug-forming group 4-aminoquinazoline knot in the surface recognition region of the compound.

It is well known that targeted drugs have certain specificities. They cannot directly obtain another series of compounds with corresponding activities or activities due to simple modification of a structure in a compound. It is necessary to consider the difference between the structure of the compound and the spatial conformation of the target protein and the key. Binding sites, design, synthesis and biological screening to obtain a series of active new derivatives. The invention obtains a series of new compounds of 4-aminoquinazoline hydroxamic acid by a large number of experimental designs and active screening, and finds active compounds with better antitumor activity and low toxicity, and has intensive research. value. It is also an advantage of the present invention 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. The tumor is a solid tumor and a hematoma, preferably liver cancer, lung cancer, breast cancer, esophageal cancer, gastric cancer, nasopharyngeal cancer, ovarian cancer, bladder cancer, rectal cancer, skin cancer, and lymphoma. In summary, the compound of the present invention has less toxic side effects as an antitumor drug, and is more easily used as an antitumor drug, which is equivalent to the prior art, and the present invention has novel, creative and scientific advances. 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. Example 1

Synthesis of V -l N-hydroxy-7-(quinazolin-4-amino)heptanamide

4-chloroquinazoline (0.164 g, 1 mmol), 7-aminoheptanoic acid methyl ester hydrochloride (0.21 g, 1 mmol), KOH (0.14 g, 2.5 mmol), hydroxylamine hydrochloride (0.7 g, 2.5 mmol) Raw materials, according to

The V-type compound was synthesized and synthesized by V-l, and the yield was 55.9%.

^SI-MS [M+H] ⁺ : m/z 289.1695

1H hire R (400MHz, DMSO-d ₆ ) 5ppm:

1.30 (m, 4H), 1.48 (m, 2H), 1.60 (m, 2H), 1.70- 1.80

(m, 3H), 1.95 (t, lH, J = 8.0 Hz), 3.52 (m, 2H), 7.49 (t, lH, J = 8.0 Hz), 7.66 (d, l H, J = 8.0 Hz), 7.75 (t, lH, J = 8.0 Hz), 8.27 (d, lH, J = 8.0 Hz), 8.34 (s, lH), 8.43 (s, lH) Example 2

Synthesis of V -2 N-hydroxy-7-(6,7-dimethoxyethoxyquinazolin-4-amino)heptanamide

6,7-Dimethoxyethoxy-4-chloroquinazoline (0.312 g, 1 mmol), 7-aminoheptanoic acid methyl ester hydrochloride (0.21 g, 1 mmol), KOH (0.14 g, 2.5 mmol Hydroxylamine hydrochloride (0.7 g, 2.5 mmol) was used as a raw material to synthesize V-2 according to the synthesis of compound V, with a yield of 65.3%. ^SI-MS [M+H] 十: m/z 437.2449

1H NMR (400MHz, DMSO-d ₆ ) 5ppm:

1.32(m,4H), 1.42- 1.63(m,4H), 1.96(t,2H,J=8.0

Hz), 2.21 (t, lH, J = 8.0 Hz), 3.34 - 3.51 (m, 7H), 3.73 (s, 6H), 4.25 (m, 4H), 7.1 5 (S, 1H), 7.91 (S, 1H), 8.38 (S, 1H), 8.64 (S, 1H) Example 3

Synthesis of V -3 N-Hydroxy-7-(6,7-dimethoxyquinazolin-4-amino)heptanamide

6,7-Dimethoxy-4-chloroquinazoline (0.224 g, 1 mmol), 7-aminoheptanoic acid methyl ester hydrochloride (0.21 g, 1 mmol), KOH (0.14 g, 2.5 mmol) Hydroxylamine (0.7 g, 2.5 mmol) was used as a raw material, and V-3 was synthesized according to the synthesis of a compound of the V type, and the yield was 66.0%.

^SI-MS [M+H] ⁺ : m/z 349.32

1H NMR (400MHz, DMSO-d ₆ ) 5ppm:

1.32(m,4H), 1.49(m,2H), 1.61 (m,2H), 1.69(s,

lH), 1.96 (t, 2H, J = 8.0 Hz), 3.48-3.57 (m, 3H), 3.88 (s, 6H), 7.06 (s, lH), 7.66 (S, 1H), 8.06 (S, 1H) ), 8.32 (S, 1H) Example 4

Synthesis of V -4 N-Hydroxy-6-(quinazolin-4-amino)hexanamide

4-chloroquinazoline (0.164 g, 1 mmol), 6-aminohexanoic acid methyl ester hydrochloride (0.196 g, 1 mmol), KOH (0.14 g, 2.5 mmol), hydroxylamine hydrochloride (0.7 g, 2.5 mmol) Raw materials, according to

V-type compound synthesis synthesis V -4, yield 50%.

^SI-MS [M+H] ⁺ : m/z 275.1547

1H NMR (400MHz, DMSO-d ₆ ) 5ppm:

1.33(m,2H), 1.4- 1.69(m,4H), 1.96(t,2H,J=8.0

Hz), 3.39 (s, lH), 3.52 (m, 2H), 7.50 (t, lH, J = 8.0 Hz), 8.23-8.28 (m, 2H), 8.4 5 (s, lH) Example 5

Synthesis of V -5 N-Hydroxy-6-(6,7-dimethoxyquinazolin-4-amino)hexanamide

6,7-Dimethoxy-4-chloroquinazoline (0.224 g, 1 mmol), 6-aminohexanoic acid methyl ester hydrochloride (0.196 g, 1 mmol), KOH (0.14 g, 2.5 mmol), hydrochloric acid Hydroxylamine (0.7 g, 2.5 mmol) was used as a raw material, and V-5 was synthesized according to the synthesis of a compound of the V type, and the yield was 66.3%.

^SI-MS [M+H] ⁺ : m/z 335.32

1H hire R (400MHz, DMSO-d ₆ ) 5ppm:

1.34(m,2H), 1.52-1.70(m,4H), 1.97(t,2H,J=8.0

Hz), 3.37 (s, 2H), 3.50 (m, 2H), 3.89 (s, 6H), 7.08 (s, lH), 7.59 (s, lH), 7.91 (t, l

H, J = 4.0 Hz), 8.33 (s, lH) Example 6

Synthesis of V -6 N-Hydroxy-7-(6-methylquinazolin-4-amino)heptanamide

6-Methyl-4-chloroquinazoline (0.178 g, 1 mmol), 7-aminoheptanoic acid methyl ester hydrochloride (0.21 g, 1 mmol), KOH (0.14 g, 2.5 mmol), hydroxylamine hydrochloride (0.7 g , 2.5 mmol) as raw material, V-6 was synthesized according to the synthesis of compound V, with a yield of 59.3%.

^SI-MS [M+H] ⁺ : m/z 303.31

1H hire R (400MHz, DMSO-d ₆ ) 5ppm:

I.32(m,4H), 1.50(m,2H),l.61 (m,2H),l.97(m,

2H), 2.46 (s, 3H), 3.51 (m, 2H), 7.57 (t, 2H, J = 8.0 Hz), 8.04 (s, lH), 8.12 (t, lH, J = 8.0 Hz), 8.40 ( s,lH) Example 7

Synthesis of V -7 N-Hydroxy-7-(6-chloroquinazolin-4-amino)heptanamide

6-Chloro-4-chloroquinazoline (0.197 g, 1 mmol), 7-aminoheptanoic acid methyl ester hydrochloride (0.21 g, 1 mmol), KOH (0.14 g, 2.5 mmol), hydroxylamine hydrochloride (0.7 g, 2.5 mmol) as raw material, according to

The V compound was synthesized by the synthesis of V-7, and the yield was 44.9%. ^SI-MS [M+H] 十: m/z 323.28

1H NMR (400MHz, DMSO-d ₆ ) 5ppm:

1.33(m,4H), 1.49(m,2H), 1.61 (m,2H), 1.95(m,2H

), 3.39 (m, lH), 3.51 (m, 2H), 7.68 (d, lH, J = 8.0 Hz), 7.77 (d, lH, J = 8.0 Hz), 8. 34 (t, lH, J = 8.0 Hz), 8.42 (s, lH), 8.47 (s, lH) Example 8

Synthesis of V -8 N-Hydroxy-7-(6-fluoroquinazolin-4-amino)heptanamide

6-fluoro-4-chloroquinazoline (0.182 g, 1 mmol), 7-aminoheptanoic acid methyl ester hydrochloride (0.21 g, 1 mmol), KOH (0.14 g, 2.5 mmol), hydroxyamine hydrochloride (0.7 g, 2.5 mmol) was used as a raw material to synthesize V -8 according to the synthesis of compound V, with a yield of 63.0%.

^SI-MS [M+H] ⁺ : m/z 307.31

1H NMR (400MHz, DMSO-d ₆ ) 5ppm:

1.33(m,4H), 1.50(m,2H), 1.61 (m,2H), 1.90(s,

lH), 1.95 (d, 2H, J = 8.0 Hz), 3.35 (m, lH), 3.50 (m, 2H), 7.66 (m, lH), 7.75 (m, lH), 8.1 1 (m, lH) , 8.20 (t, lH, J = 4.0 Hz), 8.46 (s, lH) Example 9

Synthesis of V -9 N-hydroxy-7-(7-chloroquinazolin-4-amino)heptanamide

7-Chloro-4-chloroquinazoline (0.197 g, 1 mmol), 7-aminoheptanoic acid methyl ester hydrochloride (0.21 g, 1 mmol), KOH (0.14 g, 2.5 mmol), hydroxylamine hydrochloride (0.7 g, 2.5 mmol) was used as a raw material to synthesize V-9 according to the synthesis of compound V, with a yield of 48.6%.

^SI-MS [M+H] ⁺ : m/z 323.28

1H NMR (400MHz, DMSO-d ₆ ) 5ppm:

1.32(m,4H), 1.50(m,2H), 1.62(m,2H), 1.94(t,2H,

J = 8.0 Hz), 3.35 (m, 2H), 3.50 (m, 2H), 7.55 (m, lH), 7.70 (m, lH), 8.28 (d, lH, J = 12.0 Hz), 8.41 (t, lH, J=4.0Hz), 8.46(s,lH) Example 10

Synthesis of V -10 N-Hydroxy-7-(6-nitroquinazolin-4-amino)heptanamide

6-Nitro-4-chloroquinazoline (0.209 g, 1 mmol), 7-aminoheptanoic acid methyl ester hydrochloride (0.21 g, 1 mmol), KOH (0.14 g, 2.5 mmol), hydroxylamine hydrochloride (0.7 g , 2.5 mmol) as raw material, V-10 was synthesized according to the synthesis of compound V, with a yield of 51.2%.

^SI-MS [M+H] ⁺ : m/z 335.13

1H hire R (400MHz, DMSO-d ₆ ) 5ppm:

1.33(m,4H), 1.51 (m,2H), 1.66(m,2H), 1.95(t,2H,

J = 8.0 Hz), 3.17 (d, lH, J = 8.0 Hz), 3.35 (m, 2H), 3.55 (m, 2H), 7.82 (d, lH, J = l 2.0 Hz), 8.47 (m, lH) ), 8.59 (s, lH), 8.68 (s, lH) Example 1 1

Synthesis of V -l l N-Hydroxy-7-(6-aminoquinazolin-4-amino)heptanamide

V -10 (0.333 g, 1 mmol) was dissolved in methanol, 0.04 g of 5% palladium on carbon was added, hydrogen was introduced under normal temperature and normal pressure, and stirred for 3 hours. After completion of the reaction, the palladium on carbon was removed by filtration, and the filtrate was concentrated to obtain a product.

V -11 , yield 95.0%.

^SI-MS [M+H] 十: m/z 304.16

1H hire R (400MHz, DMSO-d ₆ ) 5ppm:

1.30(m,4H), 1.48(m,2H), 1.60(m,2H), 1.70- 1.79

(m, 3H), 1.95 (t, lH, J = 8.0 Hz), 3.52 (m, 2H), 6.27 (s, 2H), 7.49 (t, lH, J = 8.0 Hz), 7.66 (d, lH) , J = 8.0 Hz), 7.75 (t, lH, J = 8.0 Hz), 8.27 (d, lH, J = 8.0 Hz), 8.34 (s, lH), 8.43 (s, lH) Example 12

Synthesis of V -12 Ν-hydroxy-7-(6-cyanoquinazolin-4-amino)heptanamide

6-Cyano-4-chloroquinazoline (0.189 g, 1 mmol), 7-aminoheptanoic acid methyl ester hydrochloride (0.21 g, 1 mmol), KOH (0.14 g, 2.5 mmol), hydroxylamine hydrochloride (0.7 g , 2.5 mmol) as raw material, V-12 was synthesized according to the synthesis of compound V, with a yield of 50.6%. ^SI-MS [M+H] 十: m/z 314.16

1H NMR (400MHz, DMSO-d ₆ ) 5ppm:

1.32(m,4H), 1.51 (m,2H), 1.66(m,2H), 1.97

(t, 2H, J = 8.0 Hz), 3.17 (d, lH, J = 8.0 Hz), 3.35 (m, 2H), 3.55 (m, 2H), 7.82 (d, l H, J = 12.0 Hz), 8.47 (m, lH), 8.59 (s, lH), 8.71 (s, lH) Example 13

Synthesis of V -13 N-Hydroxy-7-(6-acetamidoquinazolin-4-amino)heptanamide

6-Acetyl-4-chloroquinazoline (0.221 g, 1 mmol), 7-aminoheptanoic acid methyl ester hydrochloride (0.21 g, 1 mmol), KOH (0.14 g, 2.5 mmol), hydroxylamine hydrochloride (0.7 g , 2.5 mmol) as raw material, V-13 was synthesized according to the synthesis of compound V, with a yield of 69.2%.

^SI-MS [M+H] ⁺ : m/z 346.17

1H NMR (400MHz, DMSO-d ₆ ) 5ppm:

1.30 (m, 4H), 1.51 (m, 2H), 1.66 (m, 2H), 1.97

(t, 2H, J = 8.0 Hz), 2.14 (s, 3H), 3.17 (d, lH, J = 8.0 Hz), 3.35 (m, 2H), 3.55 (m, 2 H), 7.23 (s, lH) ), 7.82 (d, lH, J = 12.0 Hz), 8.47 (m, lH), 8.59 (s, lH), 8.71 (s, lH) Example 14

Synthesis of V -14 N-Hydroxy-7-(8-chloroquinazolin-4-amino)heptanamide

4,8-Dichloroquinazoline (0.221 g, 1 mmol), 7-aminoheptanoic acid methyl ester hydrochloride (0.21 g, 1 mmol), KOH (0.14 g, 2.5 mmol), hydroxylamine hydrochloride (0.7 g, 2.5 Mmmol) as raw material, according to

The V-type compound was synthesized and synthesized by V-13, and the yield was 69.2%.

^SI-MS [M+H] ⁺ : m/z 323.27

1H NMR (400MHz, DMSO-d ₆ ) 5ppm:

1.31 (m, 4H), 1.50 (m, 2H), 1.64 (m, 2H), 1.94 (t, 2H,

J = 8.0 Hz), 3.36 (m, 2H), 3.50 (m, 2H), 7.54 (m, lH), 7.70 (m, lH), 8.27 (d, lH, J = 12.0 Hz), 8.41 (t, lH, J=4.0Hz), 8.49(s,lH) Example 15

Synthesis of V -15 N-Hydroxy-7-(5-fluoroquinazolin-4-amino)heptanamide

5-Fluoro-4-chloroquinazoline (0.182 g, 1 mmol), 7-aminoheptanoic acid methyl ester hydrochloride (0.21 g, 1 mmol), KOH (0.14 g, 2.5 mmol), hydroxylamine hydrochloride (0.7 g, 2.5 mmol) as raw material, according to

Synthesis of V -15 by V compound, the yield was 45.9%.

^SI-MS [M+H] ⁺ : m/z 307.16

1H NMR (400MHz, DMSO-d ₆ ) 5ppm:

1.34(m,4H), 1.50(m,2H), 1.64(m,2H), 1.94(t,2H,

J = 8.0 Hz), 3.33 (m, 2H), 3.50 (m, 2H), 7.54 (m, lH), 7.70 (m, lH), 8.29 (d, lH, J = 12.0 Hz), 8.48 (t, lH, J = 4.0 Hz), 8.66 (s, lH) Example 16

Synthesis of V -16 N-Hydroxy-7-(2-chloroquinazolin-4-amino)heptanamide

2,4-Dichloroquinazoline (0.221 g, 1 mmol), 7-aminoheptanoic acid methyl ester hydrochloride (0.21 g, 1 mmol), KOH (0.14 g, 2.5 mmol), hydroxylamine hydrochloride (0.7 g, 2.5 Mmmol) as raw material, according to

The V-type compound was synthesized and synthesized by V-16, and the yield was 69.3%. ^SI-MS [M+H] 十: m/z 323.27

1H NMR (400MHz, DMSO-d ₆ ) 5ppm:

1.35 (m, 4H), 1.51 (m, 2H), 1.76 (m, 2H), 1.94 (t, 2H,

J = 8.0 Hz), 3.36 (m, 2H), 3.53 (m, 2H), 7.44 (m, lH), 7.70 (m, lH), 8.21 (d, lH, J = 12.0 Hz), 8.41 (t, lH, J = 4.0 Hz), 8.55 (s, lH) Example 17

Inhibitory activity of histone deacetylase in vitro

The inhibitory activity of the compound on HDACs was tested using the K340-100 kit from Biovision. The experimental procedure was carried out according to the kit instructions. First test Ι μΜ and Ο. Ι μΜ at two concentrations, the compound inhibits the percentage of the enzyme, the compound with better activity continues the inhibitory IC _{5 ()} test. The experimental results are shown in Table 2. Table 2 Experimental results of in vitro inhibitory activity of compounds on HDACs

HDACs % Inhibition Rate HDACs 1 μΜ 0.1 μΜ IC ₅₀ (nM)

SAHA 42.96

V-1 81.03 37.88 228.7

V-2 88.92 58.36 86.60

V-3 95.89 79.52 25.44

V-4 92.76 63.80 NT

V-5 93.78 70.75 NT

V-6 96.78 70.75 10.29

V-7 96.63 83.90 3.08

V-8 92.91 63.59 43.02

V-9 95.56 77.52 21.98

V-10 94.84 74.60 29.83

V-11 89.97 63.21 NT

V-12 68.90 20.83 NT

V-13 92.18 75.75 22.89

V-14 90.08 71.99 79.63

V-15 89.63 53.90 NT

V-16 92.91 61.59 55.96 As can be seen from Table 2 above, the compounds of the present invention have strong inhibitory activity against HDACs, and some compounds such as V-3, V-6, V-7, V-9, V-10, V -13 HDACs inhibitory activity is superior to the positive control drug SAHA. Example 18

In vitro inhibitory activity assay of tumor cells of the compounds of the invention:

Determination of the compounds of the invention against Jurkat E6-1 (human T cell lymphoma), Hut78 (T lymph Baine cell leukemia cells), Colo320 (human rectal cancer cell line), Caki-1 (human renal cell carcinoma cell line), MDA-MB-435S (human breast cancer cell), A549 (human lung cancer cell), PANC-1 ( The activity of human pancreatic cancer cells, IC _5Q value was measured by CCK-8 method (Cat# CK04-13, Dojindo), and SAHA was selected as a control drug. The specific results are shown in Table 3 (unit: μΜ): Table 3 In vitro inhibitory activity of the compound of the present invention on tumor cells

Num Jurkat E6-1 Hut-78 Colo320 Caki-1 435s A549 PANC-1

SAHA 0.08 0.19 2.89 4.56 6.76 0.98 10.6

V-l 6.66 5.43 52.0 3.72 20.3 26.5 29.8

V-2 2.25 2.86 35.7 2.53 26.9 3.72 6.73

V-3 0.25 0.46 30.7 0.28 24.8 2.39 2.78

V-4 0.06 0.04 32.5 0.32 20.2 0.89 0.62

V-5 3.33 3.04 30.78 3.44 39.7 39.7 30.9

V-6 0.93 3.43 3.76 0.35 0.36 0.98 3.44

V-7 0.04 0.05 7.63 0.30 8.99 0.53 6.54

V-8 0.33 0.05 3.33 0.37 3.37 3.43 7.63

V-9 0.08 0.33 3.04 0.50 0.35 0.35 3.78

V-10 0.07 0.05 3.33 0.66 30.7 0.37 50

V-ll 0.13 0.07 3.63 0.45 33.5 1.44 7.89

V-12 0.05 0.13 1.78 0.76 0.07 0.57 1.78

V-13 0.04 0.09 4.61 0.03 50 0.43 5.43

V-14 0.04 0.05 1.88 0.13 1.71 0.46 0.88

V-15 0.09 0.01 0.66 0.22 50 1.77 0.43

V-16 0.07 0.10 6.70 0.09 50 0.82 2.53 As can be seen from Table 3 above, some of the compounds of the present invention tested were inhibited against Jurkat E6-1 (human T cell lymphoma), Hut78 (T lymph) relative to the positive control drug SAHA. Cell leukemia cells), Colo320 (human rectal cancer cell line), Caki-1 (human renal cell carcinoma cell line), MDA-MB-435S (human breast cancer cell), A549 (human lung cancer cell), PANC-1 (human) Pancreatic cancer cells) have better activity in tumor cell proliferation, indicating the 4-amino group of the present invention The quinazoline hydroxamic acid compounds have good inhibitory activity against various tumor cells. Example 19

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 measured, and the IC ₅ o value was measured by the CCK-8 method (Cat# CK04-13, Dojindo). The S AHA was selected as a control drug for the in vitro inhibitory activity 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

SAHA 1.25

V-2 9.9

V-3 6.9

V-4 2.5

V-5 5.9

V-6 7.9

V-7 2.2

V-8 9.7

V-9 4.3

V-10 8.6

V-12 4.6

V-13 7.7

V-14 6.9

V-15 8.4

V-16 6.4 It can be seen from Table 4 that the compounds of the present invention V-1 to V-30 are relative to the control drug SAHA has weak inhibitory activity against normal cells and has lower toxic side effects, indicating that the 4-aminoquinazoline hydroxamic acid compounds of the present invention have better selectivity for inhibiting proliferation of tumor cells and normal cells. It indicates that it has lower toxic side effects when used as an anti-tumor drug, and is easy to use as a tumor drug. Example 20

Acute toxicity test: The method reported by Zhang Juntian, "Modern Pharmacological Experimental Method" (Beijing Medical University, China Union Medical University, United Press, 1998), preliminary screening, using Bliss method statistics ("Practical pharmaceutical preparations" Technology, People's Health Publishing House, published in 1999), the LD _{50 of} single V4, V-7, V-9, V-16 mice was 1.6g/kg, l.lg/kg 1.9g/kg and 1.3g/kg. Example 21

Tablets: Any compound selected from V-1 to V-16 100 mg sucrose 150 mg corn starch 38 mg calcium stearate 2 mg Preparation method: Mix any compound of active ingredients V-1 to V-16 with sucrose, corn starch, Add water to the wet, stir evenly, dry, smash and sieve, add calcium stearate, mix well, and compress. Each tablet weighs 290 mg and has an active ingredient content of 100 mg. Example 22

Injection: 15mg of any compound selected from V-1 to V-16 80mg of water for injection Preparation method: Dissolve any compound of active ingredient V-1 to V-16 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 limited by the above embodiments, and any other embodiments do not depart from the spirit and principle of the present invention. All changes, modifications, substitutions, combinations, and simplifications that are made are equivalent substitutions, and are included in the scope of the present invention.

Claims

Rights request

1. 4-aminoquinazoline hydroxamic acid compounds, characterized in that they have a chemical formula shown in (V):

(V)

Among them: , R ₂ , R ₃ , R ₄ or R ₅ are respectively hydrogen, halogen, amino, nitro, hydroxylamino, — C ₄ alkoxycarbonyl, — C 4 alkyl, — C ₄ alkyl, — amido group, alkyloxy group, guanidino group, ureido group, trifluoromethyl group, — sulfonyl group, arylsulfonyl group, substituted phenyl group, phenyl group or heterocycle; n= an integer from 0 to 5.

2. The 4-aminoquinazoline hydroxamic acid compound according to claim 1, characterized in that the substituted phenyl group contains 1 to 4 substituents on the benzene ring, and the substituents are halogen, Hydroxy, nitro, cyano, alkyloxy, alkyl or amino groups of 1 to 4 carbon atoms.

3. The 4-aminoquinazoline hydroxamic acid compound according to claim 2, characterized in that the halogen is fluorine, chlorine, bromine or iodine.

4. The 4-aminoquinazoline hydroxamic acid compound according to claim 1, characterized in that the alkyl group of C _r C ₄ is methyl, ethyl, propyl, isopropyl, cyclo Propyl, butyl, isobutyl, sec-butyl, tert-butyl or cyclobutyl.

5. The 4-aminoquinazoline hydroxamic acid compound according to claim 1, characterized in that the alkoxy group is -C ₄ alkyloxy group, substituted benzyloxy group, pyrrole alkyl-1 -base-(C ₂ -C ₄ )alkyloxy, morpholin-1-yl-(C ₂ -C ₄ )alkyloxy, piperazin-1-yl-(C ₂ -C ₄ )alkyloxy,

N-methylpiperazin-1-yl-(C ₂ -C ₄ )alkoxy or piperidin-1-yl-(C ₂ -C ₄ )alkoxy.

6. The 4-aminoquinazoline hydroxamic acid compound according to claim 1, characterized in that the aminoalkyl group is aminoethyl, 1-aminopropyl or 2-aminopropyl.

7. The 4-aminoquinazoline hydroxamic acid compound according to claim 1, characterized in that the alkyl amino group is N-methylamino group, N-ethylamino group or N-isopropylamino group.

8. The 4-aminoquinazoline hydroxamic acid compound according to claim 1, wherein the amide group is acetylamino (C¾C(O)NH-), propionylamino (CH ₃ CH ₂ C(O)NH -), butylamino or isobutyrylamino.

9. The 4-aminoquinazoline hydroxamic acid compound according to claim 1, wherein the sulfonyl group is methanesulfonyl, ethylsulfonyl, propylsulfonyl, isopropylsulfonyl, n-butyl Sulfonyl, isobutylsulfonyl or sec-butylsulfonyl.

10. The 4-aminoquinazoline hydroxamic acid compound according to claim 1, characterized in that:

The substituted phenyl group contains 1 to 4 substituents on the benzene ring, and the substituents are halogen, hydroxyl, nitro, cyano, alkyloxy, alkyl or amino groups of 1 to 4 carbon atoms;

The halogen is fluorine, chlorine, bromine or iodine;

The alkyl group of C _r C ₄ is methyl, ethyl, propyl, isopropyl, cyclopropyl, butyl, isobutyl, sec-butyl, tert-butyl or cyclobutyl;

The alkyloxy group is -C ₄ alkyloxy group, substituted benzyloxy group, pyrrole alkyl-1-yl-(C ₂ -C ₄ ) alkyloxy group, morpholin-1-yl-(C ₂ -C ₄ ) Alkyloxy, piperazin-1-yl-(C ₂ -C ₄ ) alkyloxy, N-methylpiperazin-1-yl-(C ₂ -C ₄ ) alkyloxy or piperidine-1 -base-(C ₂ -C ₄ )alkyloxy group;

The aminoalkyl group is aminoethyl, 1-aminopropyl or 2-aminopropyl;

The alkyl amino group is N-methylamino, N-ethylamino or N-isopropylamino;

The amide group is acetamido (CH ₃ C(O)NH-), propionylamino (CH ₃ CH ₂ C(O)NH -), butylamino or isobutyrylamino;

The sulfonyl group is methanesulfonyl, ethylsulfonyl, propylsulfonyl, isopropylsulfonyl, n-butylsulfonyl, isobutylsulfonyl or sec-butylsulfonyl.

11. 4-aminoquinazoline hydroxamic acid compounds, characterized in that they are selected from:

V-l N-hydroxy-7-(quinazoline-4-amino)heptanamide,

V-2 N-hydroxy-7-(6,7-dimethoxyethoxyquinazolin-4-amino)heptanamide, V-3 N-hydroxy-7-(6,7-dimethoxy Quinazoline-4-amino)heptylamide,

V-4 N-hydroxy-6-(quinazoline-4-amino)hexanamide,

V-5 N-hydroxy-6-(6,7-dimethoxyquinazoline-4-amino)hexanamide,

V-6 N-Hydroxy-7-(6-methylquinazoline-4-amino)heptanamide,

V-7 N-Hydroxy-7-(6-chloroquinazoline-4-amino)heptanamide,

V-8 N-Hydroxy-7-(6-fluoroquinazoline-4-amino)heptanamide,

V-9 N-Hydroxy-7-(7-chloroquinazoline-4-amino)heptanamide,

V-10 N-Hydroxy-7-(6-nitroquinazoline-4-amino)heptanamide,

V-l l N-hydroxy-7-(6-aminoquinazoline-4-amino)heptanamide,

V-12 N-Hydroxy-7-(6-cyanoquinazoline-4-amino)heptanamide,

V-13 N-Hydroxy-7-(6-acetamidoquinazoline-4-amino)heptylamide,

V-14 N-Hydroxy-7-(8-chloroquinazoline-4-amino)heptanamide,

V-15 N-Hydroxy-7-(5-fluoroquinazoline-4-amino)heptanamide or

V-16 N-Hydroxy-7-(2-chloroquinazoline-4-amino)heptanamide,

or a pharmaceutically acceptable salt thereof.

12. The 4-aminoquinazoline hydroxamic acid compound according to any one of claims 1 to 11, characterized in that the salt is hydrochloride, hydrobromide, sulfate, acetic acid Salt, lactate, tartrate, tannate, citrate, trifluoroacetate, malate, maleate, succinate, p-toluenesulfonic acid or mesylate.

13. Application of the 4-aminoquinazoline hydroxamic acid compound described in any one of claims 1 to 12 in the preparation of drugs for treating tumors.

14. The application according to claim 13, wherein the tumor includes a solid tumor or a hematological tumor.

15. A pharmaceutical composition, characterized by comprising a therapeutically effective amount of the 4-aminoquinazoline hydroxamic acid compound described in any one of claims 1 to 12 and a pharmaceutically acceptable carrier.