WO2005058875A1

WO2005058875A1 - Piperazine triazine compounds, preparation thereof and pharmaceutical compositions

Info

Publication number: WO2005058875A1
Application number: PCT/CN2004/001368
Authority: WO
Inventors: Hualiang Jiang; Hong Liu; Xu Shen; Xingzu Zu; Mingyue Zheng; Mingfang Zheng; Xinquan Ji; Haibin Luo; Hongxia Guo; Xiaomin Luo; Jianhua Shen; Kaixian Chen
Original assignee: Shanghai Institute Of Materia Medica, Chinese Academy Of Sciences; Shanghai Leaddiscovery Pharmaceutical Company
Priority date: 2003-12-19
Filing date: 2004-11-29
Publication date: 2005-06-30
Also published as: CN1629157A

Abstract

The invention provides piperazine triazine compounds of Formula (I) and They’re pharmaceutically acceptable salts: Formula (I) wherein: R1 is hydrogen, C1-C4alkyl, aryl, substituted aryl, aralkyl, C1-C4alkanoyl, aroyl or C3-C7cycloalkyl; R2 and R3 are respectively aryl, substituted aryl, aralkyl, C1-C4alkaryl or C3-C7cycloalkyl; X is NR7,O,S;R4 and R6 are respectively hydroxyl or C1-C6alkyl; R5 is hydroxyl, C1-C6alkyl,-C(O)R8,-(CH2)mR8,-CH2CH=CHR8,-C(O)OR8,-C(O)OR8 or S(O)2R8, m is 0,1,2 or 3; R7 is hydrogen, hydroxyl or C1-C6alkyl; R8 is hydrogen, hydroxyl, aryl, heteroaryl, heterocyclic radical or C2-C6alkenyl. It is approved that the piperazine triazine compounds of the present invention are epoxy synthase (COX-1 and COX-2) inhibitors with fine curative effect and prevention to clinic inflammation through computer virtuality screening, epoxy synthase(COX-1 and COX-2) combination experiment and pharmacological test. Preparation and pharmaceutical compositions of the compounds are also provided.

Description

Piperazine triazine compound, its preparation method and pharmaceutical composition

The present invention relates to the fields of medicinal chemistry and medicinal therapy, and particularly to piperazine triamidine compounds, a preparation method thereof, and a pharmaceutical composition containing such compounds. BACKGROUND OF THE INVENTION The pathogenesis of many diseases involves the involvement of inflammatory mediators. Some inflammatory diseases (such as rheumatoid arthritis, rheumatic fever, osteoarthritis), asthma, chronic obstructive pulmonary disease, trauma, burns, endotoxin shock, Alzheimer's disease and even heart failure are involved in inflammatory mediators. Considering the complexity and diversity of mediators, a single mediator is not the entire cause of the pathophysiology of inflammatory disorders. Therefore, compounds that can interfere with multiple mediators or enzymes at the same time may have a greater effect on inflammation than a single active compound. Among various inflammatory mediators, the metabolites of arachidonic acid (AA), prostaglandin (PGs) and leukotriene (LTs) are two important inflammatory mediators. The metabolism of AA has the following two pathways: (1) Cyclooxygenase (C0X) metabolic pathway, that is, AA is metabolized into PGs through a series of reactions under the catalysis of C0X; (2) Lipoxygenase (5-L0X) metabolism Pathway, that is, AA is metabolized into LTs under the catalysis of 5-LOX. Therefore, blocking the metabolism of M can inhibit the production of inflammatory mediators PGs and LTs, thereby achieving the purpose of treating or weakening inflammation.

Among them, the first metabolic pathway includes the following series of reactions: Membrane phospholipids produce free arachidonic acid (AA) via ttl phospholipase A ₂ (PLA ₂ ), and AA via cyclooxygenases (COX-1 and COX-2) The effect is converted into prostaglandin G ₂ (PG ₂ ), and its peroxidase activity then converts its product to prostaglandin H ₂ (PGH ₂ ). Tissue-specific isoenzymes metabolize PGH ₂ into other forms of prostaglandins Or thromboxane A ₂ (TxA ₂ ). At the same time, AA is metabolized to leukotrienes (LTs) by 5-lipoxygenase.

For the treatment of inflammation, early people used glucocorticoid anti-inflammatory drugs (SAID), but long-term use of these drugs can easily cause complications such as adrenal function decline. In 1952, Baotaisong was used in clinical practice, and the concept of non-body anti-inflammatory drug (NSAID) was first proposed in the world. In the following two or three decades, a large number of NSAIDs with anti-inflammatory and analgesic effects emerged, such as indomethacin, ibuprofen, etc., which are still widely used in clinical practice. These drugs have always led the world's pharmaceutical output. First status. However, long-term application of NSAID can cause damage to the gastrointestinal tract, especially the gastric mucosa, manifested as punctate bleeding, diffuse superficial mucosal erosion, deep local ulcers, major bleeding and even perforation.

In 1991, the discovery of COX-2 opened a new direction for NSAID research. Studies have shown that: COX-1 is expressed in normal tissues and is a constituent protein of normal cells; COX-2 is an induced form of enzymes, mainly in inflammatory cells, such as endothelial cells, macrophages, synovial fibroblasts, It is expressed in dendritic cells, chondrocytes and osteoblasts, and has similar active binding sites to COX-1 to AA or NSAID; COX-2 can be induced by various factors in inflammatory tissues, and its level will be 8-10 The rate increased sharply, causing the contents of PGE ₂ , PGI ₂ and PGE in the inflammation site to increase, which promoted the inflammatory response and tissue damage. In 1994, JR Vane et al. Pointed out that the effective therapeutic effect of NSAID on inflammation stems from its inhibition of COX-2, and the adverse reactions are attributed to the inhibition of COX-1. Therefore, since the 1990s, the search for COX-2 selective inhibitors has become an important way to discover new anti-inflammatory drugs. Many international pharmaceutical companies such as Seale, Merck, Pfizer, Glaxo-Welcome, and Tabacco have developed new COX-2 inhibitors, of which Nimesulide, Meloxicam, Celecoxib Vioxx, and ML3000 have been developed and marketed, and there are nearly 10 of them The compound is in clinical research. NSAID's new target has been proposed to speed up the discovery of anti-inflammatory drugs. Since the discovery of COX-2 in 1991, the first highly selective COX-2 inhibitor Celecoxib went on the market at the end of December 1998 and was sold in 1999. The amount actually reached 1.2 billion US dollars.

Studies have shown that there is a certain equilibrium relationship between the two metabolic pathways of AA, that is, when the activity of COX is inhibited, the activity of 5-LOX is enhanced, allowing more AA to enter the 5-LOX metabolic pathway to produce LTs. Therefore, simply inhibiting one of the metabolic pathways will cause a large number of AA to enter other metabolic pathways, which will lead to the further development of inflammation. Similarly, when the activity of 5-LOX is inhibited, more AA will enter the COX metabolic pathway to generate PG The increase, the results all make the inflammation worse, so the dual inhibitor of COX-2 / 5-LOX is designed to achieve the purpose of synergistic anti-inflammatory. Classic NSAIDs such as aspirin, indomethacin, and diclofenac are selective inhibitors of COX and generally do not affect 5-LOX metabolism. They not only affect the synthesis of PGs that have protective effects on the gastric mucosa, but also because A single inhibition of COX-2 resulted in increased LOX metabolic activity, caused an AA metabolic imbalance, promoted an increase in the synthesis of LTs substances, thereby promoting the chemotactic aggregation of white blood cells and increasing vascular permeability. Therefore, Dr. Vane proposed: A full-functioning anti-inflammatory drug with low toxicity and side effects, which should simultaneously inhibit COX and 5-LOX. In fact, the research on dual COX-2 / 5-LOX inhibitors has been the focus of research on anti-inflammatory drugs by medical workers at home and abroad in recent years.

In recent years, Computer-Aided Drug Design (CADD) has become an important method and tool for modern drug research and development. Computer-assisted drug design, especially computer-assisted combinatorial chemical library design, has been incorporated into new drug research. Recycling can shorten the cycle of new drug research, save research and development costs, and improve the hit rate of new drug screening. The combination of computer-aided drug design and other modern drug research methods will promote the rapid development of disciplines related to pharmacy and life sciences. The CADD method has been widely used in the discovery and optimization of lead compounds, and some large international pharmaceutical companies have given full attention to the application of this technology in the research of new drugs. After many years of efforts, a considerable number of computer-aided drug design drugs have been launched, such as the carbonic anhydride of Merck Sharp & Dohme (Harlow, UK). Enzyme inhibitors Dorzolamid, Roche (Welwyn, UK) company HIV protease inhibitor Saquinavir, Biota (Melbourne, Australia) company neuraminidase inhibitor Relenza and Roche (Basel, Switzerland) company thrombin inhibitor Ro466240, etc .; The highly selective COX-2 inhibitor Celecoxib developed by Searle is also based on the results of three-dimensional quantitative structure-activity relationship (3D-QSAR) research. "Combinatorial Chemistry" is a new technology that appeared in medicinal chemistry and synthetic chemistry in recent years. It can quickly generate a large number of molecular structures for high-throughput screening. Once the emergence of combinatorial chemistry methods, There is widespread concern. The discovery and optimization of lead compounds has always been a difficult and important point in medicinal chemistry research. Especially in the past one or two decades, great progress has been made in biology and other related high and new technologies, resulting in high-throughput screening technology based on cell and molecular level, which has also made the synthesis of a large number of compounds of new structure types a bottleneck in drug research. The advent of combinatorial chemistry offers huge possibilities to solve this problem. Therefore, "combined chemistry and high-throughput screening" has become the core technology of another drug research after CADD. "Structural biology" has also entered the applied research phase from the original basic research. One of the main application fields is to study the interaction between drugs and target proteins at the molecular and atomic structure level, and to determine the crystal structure of drug-protein complexes. The design of new compounds and the structural modification of lead compounds provide useful structural information. In recent years, research in this area has progressed rapidly, and structural biology techniques and methods have been successfully applied in the research of anti-AIDS drugs, anti-cold virus drugs and anti-cancer drugs. Human genome sequencing has been completed and combined with bioinformatics, structural biology will play an important role in the discovery of new targets for drug action.

The inventors comprehensively used the three-dimensional structures of COX-1, COX-2 and 5-LOX (including COX-1 and COX-2 as crystal structures, and the three-dimensional structure of 5-L0X using homologous protein modeling method). Computer-assisted drug molecular design, combinatorial chemistry, molecular biology, and structural biology methods to find lead compounds with dual inhibitors of COX-2 / 5-LOX and optimize their structure for their pharmacological effects. The inventors have found piperazine-triazine compounds having a new structure for a good combination C0X-2 enzymes, the dissociation constant of ^10-6 and mouse carrageenan paw edema model has a good anti-inflammatory effect, thus completing the this invention. Summary of the invention

An object of the present invention is to provide a piperazinetriazine compound having a dual inhibitory effect on COX-2 / 5-LOX.

Another object of the present invention is to provide a method for synthesizing piperazine triazine compounds by using trichloromethane as a raw material.

Another object of the present invention is to provide a pharmaceutical composition for treating inflammatory diseases containing a piperazinetriazine compound. The present invention provides a piperazine triazine compound or a pharmaceutically acceptable salt thereof having the structure of the following general formula (I):

Work,

R is H, dC ₄ fluorenyl, aryl, substituted aryl, arylfluorenyl, alkanoyl, aroyl, or C ₃ -C ₇ cyclofluorenyl;

Is aryl, substituted aryl, arylfluorenyl, C C4 alkylaryl or c ₃ -c ₇ cyclofluorenyl;

X is NR ₇ , 0 or S;

R ₃ is aryl, substituted aryl, arylfluorenyl, fluorenylaryl or C ₃ -C ₇ cycloalkyl;

R4 is hydroxy or ^-^ fluorenyl;

Is hydroxyl, Cr amidino, -C (0) R ₈ ,-(CH ₂ ) _m R ₈ , -CH ₂ CH = CHR ₈ , -C (0) OR ₈ or -S (0) ₂ R ₈ , m Is 0, 1, 2 or 3;

R ₆ is hydroxyl or -fluorenyl;

R ₇ is hydrogen, hydroxy or fluorenyl;

R ₈ is hydrogen, hydroxy, aryl, heteroaryl ring, heterocyclyl or C2-C6 alkenyl hydrocarbon.

Specific examples of the "pharmaceutically acceptable salts" in this specification include propionic acid, oxalic acid, malonic acid, succinic acid, fumaric acid, maleic acid, lactic acid, malic acid, tartaric acid, citric acid, and the like. Salts formed with organic acids and acidic amino acids such as aspartic acid and glutamic acid and then formed with inorganic bases, such as sodium, potassium, calcium, aluminum salts, and phosphonium salts, or salts formed with organic bases, such as methylamine salts , Ethylamine salts, ethanolamine salts, etc., or salts of inorganic acids such as hydrochloric acid, hydrobromic acid, hydrofluoric acid, sulfuric acid, nitric acid, and phosphoric acid after forming esters with basic amino acids such as lysine, arginine, and ornithine , Or salts with organic acids such as formic acid, acetic acid, picric acid, methanesulfonic acid, and ethanesulfonic acid.

A preferred embodiment of the compound of formula (I) of the present invention is the following piperazinetriazine compound or a pharmaceutically acceptable salt thereof:

Wherein, when X is NR ₇ and R ₇ is hydrogen,

Ri is H, d-Ct alkyl, aryl, substituted aryl, aralkyl, d-alkanoyl, aroyl or C ₃ -C ₇ cyclofluorenyl;

Is aryl, substituted aryl, arylfluorenyl, alkaryl or ¾ ₇ cycloalkyl;

Is aryl, substituted aryl, arylfluorenyl, alkylaryl or C ₃ -C ₇ cyclofluorenyl;

R4 is hydroxy or ^-^ fluorenyl; R ₅ is hydroxyl, CrQs alkyl, -C (0) R ₈ ,-(CH ₂ ) _m R ₈ , -CH ₂ CH = CHR ₈ , -C (0) OR ₈ or -S (0) ₂ R ₈ , m is 0, 1, 2 or 3;

R ₆ is hydroxy or-^ alkyl;

It is hydrogen, hydroxy, aryl, heteroaryl ring, heterocyclyl or C2-C6 alkenyl hydrocarbon.

Another preferred embodiment of the compound of formula (I) of the present invention is the following piperazine triazine compound or a pharmaceutically acceptable salt thereof: '

Wherein, when X is NR ₇ and R ₇ is a hydroxyl group,

, H, d-fluorenyl, aryl, substituted aryl, arylfluorenyl, d-fluorenyl, aroyl, or C ₃ -C ₇ cycloalkyl;

R ₂ is aryl, substituted aryl, aralkyl, C, -C ₄ alkylaryl or C ₃ -C ₇ cyclofluorenyl;

Is aryl, substituted aryl, arylfluorenyl, C fluorenyl or c ₃ -c ₇ cyclofluorenyl;

RA is hydroxy or ^-^ alkyl;

R ₅ is hydroxyl, Ci-fluorenyl, -C (0) R ₇ ,-(CH ₂ ) _m R ₇ , -CH ₂ CH = CHR ₇ , -C (0) OR ₇ or -S (0) ₂ R ₇ , m is 0, 1, 2 or 3;

R ₆ is hydroxy or ^-^-;

Yet another preferred embodiment of the compound of formula (I) of the present invention is the following piperazinetriazine compound or a pharmaceutically acceptable salt thereof:

Among them, when X is NR ₇ ,! ₇ for. _1- . ₆ hours

, H, -C4 alkyl, aryl, substituted aryl, arylfluorenyl, fluorenyl, aroyl, or C ₃ -C ₇ cycloalkyl;

R ₂ is aryl, substituted aryl, arylfluorenyl, C _r C ₄ alkylaryl, or C ₃ -C ₇ cyclofluorenyl;

¾ is an aryl group, a substituted aryl group, an arylfluorenyl group, a CC ₄ fluorenyl group, or a C ₃ -C ₇ cyclofluorenyl group;

Is hydroxy or CC ₆ alkyl;

R ₅ is hydroxyl, -C ₆ fluorenyl, -C (0) R ₈ ,-(CH ₂ ) _m R ₈ , -CH ₂ CH = CHR ₈ ,-C (0) OR ₈ or -S (0) ₂ R ₈ , m is 0, 1, 2 or 3;

R ₆ is hydroxy or CC ₆ alkyl;

Wherein, when X is 0,

R is H, C fluorenyl, aryl, substituted aryl, aralkyl, -fluorenyl, aroyl or ¾-. ₇ Huanyuanji; R ₂ is aryl, substituted aryl, arylfluorenyl, -alkaryl or C ₃ -C ₇ cyclofluorenyl;

Is aryl, substituted aryl, arylfluorenyl, -fluorenylaryl or C ₃ -C ₇ cycloalkyl;

RA is hydroxyl or ₆ -fluorenyl;

Is hydroxy, C, -C ₆ fluorenyl, -C (0) R ₈ ,-(C¾) _m R ₈ , -C¾CH = CHR ₈ , -C (0) OR ₈ or -S (0) ₂ R ₈ , m is 0, 1, 2 or 3;

R ₆ is hydroxy or-. ₆垸基;

Another preferred embodiment of the compound of formula (I) of the present invention is the following piperazinetriazine compound or a pharmaceutically acceptable salt thereof:

Where X is S,

R, is H, d- embankment group, an aryl group, a substituted aryl group, an aryl group embankment, dC ₄ embankment acyl, aroyl, or C ₃ -C ₇ cycloalkyl group homes;

Is aryl, substituted arylfluorene, aralkyl, -fluorenylaryl or C ₃ -C ₇ cyclofluorenyl;

R ₃ is aryl, substituted aryl, arylfluorenyl, C-alkaryl or C ₃ -C ₇ cyclofluorenyl;

F is hydroxy or ₆ alkyl;

R ₅ is hydroxyl, -fluorenyl, -C (0) R ₈ ,-(CH ₂ ) _ni R ₈ , -CH ₂ CH = CHR ₈ , -C (0) OR ₈ or -S (0) ₂ R ₈ , m is 0, 1, 2 or 3:

R ₆ is hydroxy or -¾ 垸;

The present invention also provides a triazine compound represented by the following formula (II), wherein the compound of the formula (II) is an intermediate for preparing the compound of the formula (I).

In the formula, R is fluorenyl, aryl, substituted aryl, arylfluorenyl, fluorenyl, aroyl or ^-. ₇ cycloalkyl;

R ₂ is aryl, substituted aryl, arylfluorenyl, CrC ₄ fluorenyl or C ₃ -C ₇ cycloalkyl;

X is NR ₇ , 0, S;

R ₃ is aryl, substituted aryl, arylfluorenyl, fluorenyl or C ₃ -C ₇ cyclofluorenyl;

Y is hydrogen, halogen or -C4 amide;

The present invention also provides a triazine compound represented by the following formula (III), wherein the compound of the formula (III) is an intermediate for preparing the compound of the formula (I).

In the formula, R is 11, Ci-C ₄ fluorenyl, aryl, substituted aryl, aralkyl, Cr C ₄ acyl, aroyl or C ₃ -C ₇ cycloalkyl;

2 is aryl, substituted aryl, arylfluorenyl, -alkaryl or C ₃ -C ₇ cyclofluorenyl;

Y is hydrogen, halogen, amino Cr fluorenyl;

Z is hydrogen, halogen, amino C, -C ₄ fluorenyl; The present invention provides the following preparation method of a piperazine triazine compound represented by formula (I) or a pharmaceutically acceptable salt thereof:

R is H, fluorenyl, aryl, substituted aryl, aralkyl, -fluorenyl, aroyl or C ₃ -C ₇ cyclofluorenyl;

R ₂ is aryl, substituted aryl, arylfluorenyl, dC ₄ alkylaryl or C ₃ -C ₇ cyclofluorenyl;

X is NR ₇ , 0 or S;

F is hydroxyl or ₆ -fluorenyl;

R ₅ is hydroxy, C, -C ₆ fluorenyl, -C (0) R ₈ ,-(CH ₂ ) _m R ₈ , -CH ₂ CH = CHR ₈ , -C (0) OR ₈ or -S (0 ) ₂ R ₈ , m is 0, 1, 2 or 3;

6 is hydroxy or ₆ fluorenyl;

R ₇ is hydrogen, hydroxy or CrQs alkyl;

Is hydrogen, hydroxyl, aryl, heteroaryl ring, heterocyclyl or C2-C6 alkenyl hydrocarbon;

The method mainly includes the following steps: (1) Combining the formula (IV)

(Where M is hydrogen, halogen, amino dC ₄ alkyl; Y, Z are the same as defined in general formula (1), (11), (III)), in a basic solvent at a temperature of 20-100 ° C The following reaction with a substituted amine gives the compound represented by the general formula (III):

(III)

(In the formula, R ₂ , R ₂ , Y, and Z are the same as defined in the general formulae (1) and (II));

(2) reacting a compound represented by the general formula (III) with an aromatic amine, an alcohol or a thiol at a temperature of 20-100 ° C in a basic solvent to obtain a compound represented by the general formula (II):

(II)

(In the formula, R ,, R ₂ , R ₃ , X, Y are the same as defined in the general formula (I));

(3) reacting a compound represented by the general formula (II) with a substituted piperazine under a basic condition at -40 to 40 ° C to obtain a compound represented by the formula (I);

(4) If necessary, a salt formation reaction is performed.

The reaction described in step (1) is usually carried out in an inert solvent at 20-100 ° C. The solvent used may be THF, Et ₂ 0, DMF, ethylene glycol dimethyl ether, ethylene glycol diethyl ether, dioxane and the like. The base used is selected from organic bases including pyridine, triethylamine, 4-dimethylaminopyridine (DMAP), diisopropylethylamine, and inorganic bases including sodium carbonate, potassium carbonate, sodium hydroxide, potassium hydroxide .

The reaction described in step (2) is also performed in an inert solvent at 20-10 CTC. The solvent used may be THF, Et ₂ 0, DMF, ethylene glycol dimethyl ether, ethylene glycol diethyl ether, dioxane and the like. The base used is selected from organic bases including pyridine, triethylamine, 4-dimethylaminopyridine (DMAP), diisopropylethylamine, and inorganic bases including sodium carbonate: potassium carbonate, sodium hydroxide, potassium hydroxide . The polar organic solvent used in step (3) is selected from the group consisting of ethanol, methanol, ethyl acetate and tetrahydrofuran, and the base used is selected from the group consisting of pyridine, triethylamine, 4-dimethylaminopyridine (DMAP), and diisopropyl Organic bases of ethylamine and inorganic bases including sodium carbonate, potassium carbonate, sodium hydroxide, potassium hydroxide.

The products obtained in steps (1), (2), and (3) can be purified by appropriate methods such as column chromatography and recrystallization, respectively, to obtain pure products. Another aspect of the present invention relates to a pharmaceutical composition for preventing and / or treating an inflammatory disease, the pharmaceutical composition comprising a piperazine triazine compound represented by formula (I) or a pharmaceutically acceptable salt thereof and at least one pharmacy It is an acceptable carrier which can be used for in vivo treatment and is biocompatible. The pharmaceutical composition can be prepared into various forms according to different routes of administration. The pharmaceutical composition of the present invention can be used for preventing and / or treating inflammatory diseases.

The pharmaceutical composition according to the present invention which can be used for preventing and / or treating inflammatory diseases refers to an effective dose of the compound of the formula I of the present invention or a pharmaceutically acceptable salt or hydrate thereof and one or more suitable pharmacological With carrier. This pharmaceutical carrier includes, but is not limited to: ion exchangers, alumina, aluminum stearate, lecithin, serum proteins such as human albumin, buffer substances such as phosphate, glycerol, sorbic acid, potassium sorbate Partial glyceride mixtures of saturated vegetable fatty acids, water, salts or electrolytes, such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyethylene Pyrrolidone, cellulose material, polyethylene glycol, sodium carboxymethyl cellulose, polyacrylate, beeswax, lanolin.

The pharmaceutical composition of the compound of the present invention can be administered in any of the following ways: oral, spray inhalation, rectal, nasal, buccal, topical, parenteral, such as subcutaneous, intravenous, intramuscular, intraperitoneal, intrathecal , Intraventricular, intrasternal, and intracranial injections or infusions, or with the aid of an explanted reservoir. Among them, oral or intramuscular injection, intraperitoneal or intravenous administration are preferred when treating inflammation.

In addition, it should be noted that the dosage and method of use of the compounds of the present invention depend on many factors, including the age, weight, sex, natural health, nutritional status, intensity of the compound's activity, time of administration, metabolic rate, severity of the condition, and Subjective judgment of the treating physician. If the recommended dose is 5mg ~ 10mg / kg daily, the maintenance amount can be reduced to 3mg / kg per R. Capsule: 0.25g / capsule. Injection 0.25g / 5ml. Oral solution: 5g / 50ml. BRIEF DESCRIPTION OF THE DRAWINGS

Figure 1 shows the binding kinetics of piperazine triazines and cyclooxygenases (COX-1 and COX-2). The instrument used was BIACORE3000, and the analysis software was Kinetic Analysis in the Application Wizard. The cyclooxygenase 2 protein was fixed on the CM5 chip. The concentration of piperazine triazine compounds tested was: 0.625, 1.25, 2.5, 5.0, and 10.0. μM.

detailed description

The following examples further illustrate the preparation of compounds of formula (I), but these examples in no way limit the invention. Nuclear magnetic resonance spectra were measured on Bruker AM-400, and mass spectra were performed on a MAT-95 mass spectrometer. Elemental analysis was performed by the Analysis Laboratory of Shanghai Institute of Materia Medica, Chinese Academy of Sciences. The melting point is measured on an electrothermal melting point tube or a b-type melting point tube. The thermometer is not calibrated. The layer chromatography (TLC) uses silica gel GF ₂₅₄ (produced by Qingdao Ocean Chemical Plant) and a 0.8% concentration carboxymethyl cellulose sodium distillate solution. Stir well, spread the plate, dry it, activate it at 100 ~ 110 ° C for 1 ~ 2 hours, and store it in a thousand desiccator for later use. UV lamp (λ: 254 ηη) develops color; use 100 mesh to 200 mesh for column chromatography. Column chromatography silica gel (produced by Qingdao Ocean Chemical Plant).

Example 1 Synthesis of aniline monosubstituted trichlorazine

In a 50ml eggplant-shaped bottle, dissolve 4.726 g of trichlorazine in 20 ml of 1,4-dioxane, and slowly drop in an aqueous solution of 2.337 ml of aniline and 1.03 g of NaOH under stirring, and stir the reaction for 6 hours; Filtration and washing with water gave the title compound as 3.259 g, yield 53%. EA / PE = 1: 10. Example 2 Synthesis of Aniline Disubstituted Trichlorazine

In a 50 ml eggplant-shaped bottle, 0.421 g of trichlorazine was dissolved in 5 ml of 1,4-dioxane, and 0.42 ml of an aniline and 0.2 g of NaOH aqueous solution were added under stirring at room temperature; the reaction was stirred at 20-100 ° C overnight. It was filtered with suction and washed with water to obtain an off-white solid (crude product). After drying, the title compound was 0.515 g in a yield of 76% (crude). The measured melting point is 197 ° F, which is consistent with literature values. EA / PE = 1: 10 (2-3 drops of triethylamine).

Example 3 Synthesis of 2-piperazine-4,6-diphenylamine-1,3,5-triazine

In a 100ml eggplant-shaped bottle, 2.915 g of piperazine hexahydrate was dissolved in methyl ethyl ketone, and 1.117 g of aniline disubstituted trichlorochlorazine was added, and the mixture was stirred for 5 hours under _-40 and ₄ (TC and alkaline conditions (or heated to reflux) ~ 3 hours is also possible.) Stop the reaction, add cold water to precipitate a white precipitate, and suction filter. The dried white crystals were separated by column chromatography to obtain the title compound 0.702 g, yield 54%. EA / PE = 4: 3 ( Triethylamine 2-3 drops). Melting point 189 ~ 192 ° C.

'HNM (CDC1 ₃ ) ppm: δ = 2.40 (1 H, br-s, 7); 2.91 (4H, t, l); 3.83 (4H, t, 2); 7.04 (2H, 3,3); 7.10 (2H, br-s, 4); 7.30 (4H, t, 5); 7.55 (4H, d, 6).

MS (EI, m / z): 347 (M +); 291 (50%); 279 (base :).

Example 4 Synthesis of 2-aniline-4- (4'-pyridylamino) -6-chloro-1,3,5-triazine

In a 50 ml eggplant-shaped bottle, 0.325 g of aniline monosubstituted tripolychlorazine was dissolved in an appropriate amount of ethyl acetate, 0.254 g of 4-aminopyridine was added, and the reaction was carried out under stirring at room temperature for 5 hours, and then 0.525 g of a pale yellow-green crude product was suction filtered.

Example 5 Synthesis of (2-piperazine-4-aniline-6- (4'-pyridylamino) -1,3,5-triazine)

The prepared 0.525 g of crude product was dissolved in an appropriate amount of methanol to obtain a clear solution; it was slowly dropped into a large excess of piperazine hexahydrate in a methanol solution, stirred at room temperature for 0.5 hours, and then refluxed for 5 hours. After evaporating most of the methanol, a sufficient amount of water was added to obtain an emulsion. A clear white precipitate appeared when quantitative sodium chloride was added, and the crude product was obtained by suction filtration.

Example 6 Synthesis of 4-aniline-6- (4'-sulfonamidophenylamino) -1,3,5-triazine

In a 25 ml eggplant-shaped bottle, 0.250 g of aniline monosubstituted trichlorochlorazine was dissolved in acetone, and 0.179 g of sulfanilamide and 0.087 g of NaHCO ₃ were added under stirring at room temperature _{; the} reaction was refluxed for 24 hours, and part of the acetone was distilled off to precipitate a white flocculent precipitate. Jingjia half-small After suction filtration, 0.277 g of the title compound was obtained in a yield of 71%.

Example 7 Synthesis of (2-piperazine-4-aniline-6- (4'-sulfonylaminophenylamino) -1,3,5-triazine)

Except for using 4-aniline-6- (4, -sulfonylaminophenylamino) -1,3,5-triazine instead of aniline disubstituted tripolychlorazine, the procedure was carried out in Example 5 to obtain the title compound. The rate is 88%.

'HNMR (CDC1 ₃ ) ppm: δ = 2.88 (4H, t, 2); 3.84 (4H, t, 3); 7.65 (2H, d, 5); 7.29 (2H, t, 6);

7.01 (1 H, t, 7); 7.78 (2H, d, 9); 7.88 (2H, d, 10). MS (EI, m / z): 426 (M +); 358 (base).

Example 8 Synthesis of 4-aniline-6- (4, -trifluoromethoxyphenylamino) -1,3,5-triazine

The title compound was obtained in the same manner as in Example 6 except that trifluoromethoxyaniline was used instead of sulfanilamide. The yield was 78% ·. Example 9 Synthesis of 2-piperazine-4-aniline-6- (4'-trifluoromethoxyphenylamino) -1,3,5-triazine

The title compound was obtained in the same manner as in Example 5 except that 4-aniline-6- (4, -trifluoromethoxyphenylamino) -1,3,5-triazine was used instead of aniline disubstituted trichlorazine. The yield is 88%.

'HNMR (CDC1 ₃ ) ppm: δ = 3.15 (4H, t, 2); 4.00 (4H, t, 3); 7.23 (2H, d, 5); 7.23 (2H, t, 6);

7.00 (lH, t, 7); 7.75 (2H, d, 9); 7.90 (2H, d, 10) .MS (EI, m / z): 431 (M +); 375 (80%); 363 (base ).

Example 10 Synthesis of 2,4,6-triphenylamine -1,3,5-triazine

The title compound was obtained in the same manner as in Example 5 except that aniline was used instead of piperazine.

'HNMR (CDC1 ₃ ) ppm: δ = 3.15 (4H, t, 2); 4.00 (4H, t, 3); 7.23 (2H, d, 5); 7.23 (2H, t, 6); 7.00 (lH, t, 7); 7.75 (2H, d, 9); 7.90 (2H, d, 10).

MS (EI, m / z): 431 (M +); 375 (80%); 363 (base).

Example 11 Synthesis of 4-aniline-6- (4, -methylaniline) -1,3,5-triazine

The title compound was obtained in the same manner as in Example 6 except that 4-methylaniline was used in place of sulfanilamide in a yield of 60% ·.

Example 12 Synthesis of 2-piperazine-4-aniline-6- (4'-methylaniline) -1,3,5-triazine

The title compound was obtained in the same manner as in Example 5 except that 4-aniline-6- (4'-methylaniline) -1,3,5-triazine was used instead of aniline disubstituted tripolychlorazine. The yield was 64 °. /. .

'HNMR (CDC) ppm: δ = 2.92 (4H, t, 2); 3.82 (4H, t, 3); 7.14 (2H, d, 5); 7.32 (2H, t, 6); 7.02 (1 H, t, 7); 7.44 (2H, d, 9); 7.58 (2H, d, 10); 2.32 (3H, s, ll

Example 13 Synthesis of 4-aniline-6- (4'-chloroaniline) -1,3,5-triazine

The title compound was obtained in the same manner as in Example 6 except that 4-chloroaniline was used instead of sulfanilamide. The yield was 38% ·.

Example 14 Synthesis of 2-piperazine-4-aniline-6- (4'-chloroaniline) -1,3,5-triazine

The title compound was obtained in the same manner as in Example 5 except that 4-aniline-6- (4'-chloroaniline) -1,3,5-triazine was used instead of aniline disubstituted tripolychlorazine. The yield was 44%. .

'H MR (CDC1 ₃ ) ppm: δ = 2.92 (4H, t, 2); 3.80 (4H, t, 3); 7.26 (2H, d, 5); 7.32 (2H, t, 6); 7.06 (lH , t, 7); 7.51 (2H, d, 9); 7.55 (2H, d, 10).

Example 15 Synthesis of (2-piperazine-4-aniline-6- (3, -fluoroaniline) -1,3,5-triazine)

Except that 4-aniline-6- (3'-fluoroaniline) -1,3,5-triazine was used instead of aniline di-substituted trichlorazine, operation was carried out as in Example 5. This gave the title compound.

'HNMR (CDC1 ₃ ) ppm: δ = 2.81 (4H, t, 2); 3.79 (4H, t, 3); 7.75 (2H, d, 5); 7.28 (2H, t, 6); 6.98 (1 H , t, 7); 7.43 (H, dd, 9); 6.71 (H, td, 10); 7.28 (lH, s, 12).

Example 16 Synthesis of 4-aniline-6- (3'-methoxyaniline) -1,3,5-triazine

The title compound was obtained in the same manner as in Example 6 except that 3'-methoxyaniline was used in place of sulfanilamide in a yield of 83 ° /. . Example 17 Synthesis of 2-piperazine-4-aniline-6- (3'-methoxyaniline) -1,3,5-triazine

The title compound was obtained in the same manner as in Example 5 except that 4-aniline-6- (3'-methoxyaniline) -1,3,5-triazine was used instead of aniline disubstituted tripolychlorazine. The yield was 75. % '.

'HNMR (CDC1 ₃ ) ppm: δ = 2.91 (4H, t, 2); 3.80 (4H, t, 3); 7.58 (2H, d, 5); 7.30 (2H, t, 6);

7.02 (lH, t, 7); 6.84 (2H, d, 9); 7.42 (2H, d, 10).

Example 18 Synthesis of 2- (N, -p-toluenesulfonylpiperazine) -4-aniline-6-p-methoxyaniline -1,3,5-triazine

In a 25 ml eggplant-shaped bottle, 0.050 g of 4-aniline-6-p-methoxyaniline-1,3,5-triazine was dissolved in an appropriate amount of pyridine, and 0.0274 g of p-toluenesulfonyl chloride was added under stirring in an ice bath. The reaction solution turned yellow-green, stirred for 3 hours, added a sufficient amount of cold water, and filtered with suction to obtain 0.058 g of a white solid with a yield of 81%.

'HNMR (CDC) ppm: δ = 2.40 (1 H, br-s, 7); 2.91 (4H, t, l); 3.83 (4H, t, 2); 7.04 (2H, 3,3);

7.10 (2H, br-s, 4); 7.30 (4H, t, 5); 7.55 (4H, d, 6).

Test example 1

Surface Plasmon Resonance (SP) biosensor technology Biacore 3000 was used to study the interaction of piperazine triazines with cyclooxygenases (COX-1 and COX-2)

experimental method: .

Using BIACORE3000 (BIACORE AB, Uppsala, Sweden) (Amersham), binding experiments of compounds with cyclooxygenase (COX-1 and COX-2) were performed at room temperature. The chip and buffer solution are as follows: CM5 chip, EDC, NHS, ethanolamine, HBS-EP (purchased from BIACORE AB (Uppsala, Sweden)) ₀

• Operation steps: Dissolve the piperazine triazine compounds in DMSO and dilute to the corresponding concentrations (0.625, 1.25: 2.5, 5.0 and 10.0 M) with HBS-EP, and the DMSO content is 0.4%. The purified protein was connected to the chip by the amino coupling method, the compound was dissolved in DMSO, and then diluted to the required concentration with HBS-EP, and the kinetics experiments were performed using the BIACORE 3000 kinetic analysis Wizard for data collection and analysis .

The experimental results are shown in Figure 1.

Analyze the data according to the 1: 1 Langmuir binding model, and get a series of results as shown in Table 1.

Table 1.Combination of piperazine triazines with cyclooxygenase (COX-1 and COX-2)

Analysis of kinetic constants

Compound number KDcox-l KDcox-2

la 2.10E-5 8.91E-6 Test example 2 mouse carrageenan foot swelling model

The test inflammatory agent was 0.2% Carrageenin. Fifty healthy mice weighing 22 to 25 g were selected and randomly divided into groups: piperazine triazine compounds 50 mg / kg, intragastrically or intraperitoneally; indomethacin 10 mg / kg, intragastrically; negative control Groups were given the same volume of saline. Thirty minutes after a single administration, the right foot plantar of mice was injected subcutaneously with 20 μ carrageenan. Four hours later, the left and right feet were cut along the ankle joint and weighed. The administration group, the control group, and the positive group were compared. Differences in the drug control group were calculated by statistical analysis to obtain the average, SD, p-value and percent inhibition. The experimental results are shown in Table 2.

Table 2.Inhibition rates of piperazine triazine compounds on mouse carrageenan foot swelling model

Possibility of industrial use

The preparation method of the piperazinetriazine compound of the present invention has the advantages of mild reaction conditions, abundant raw materials and easy availability, simple operation and post-treatment, and the like.

The piperazine triamidine compounds of the present invention have strong anti-inflammatory activity in computer virtual screening and cyclooxygenase (COX-1 and COX-2) binding experiments and various experimental animal models, and can be used as anti-inflammatory drugs. They have a highly selective blocking effect on COX-2.

In the Biacore 3000 biosensor technology research piperazine and triazines on cyclooxygenase (COX-1 and COX-2) interaction model, the compound was observed at all concentrations in the range of 10 ^{^M-7-10-5} Both have a role in this model.

The compound of the present invention can inhibit the mouse carrageenan foot swelling model, and the intensity of the action increases with the administered dose. Compound Ic was effective by intraperitoneal injection of 6.25 mg / kg, and effective by oral administration of 25 mg / kg.

The compounds of the present invention have very low toxicity.

Therefore, the compound of the present invention can be used for preparing a medicament for treating an inflammatory disease.

Claims

Rights request

1. Piperazine triazine compounds or their pharmaceutically acceptable salts represented by the following general formula (I):

(I)

Where

R is H, dC ₄ fluorenyl, aryl, substituted aryl, arylfluorenyl, C, -C ₄ acyl, aroyl, or C ₃ -C ₇ cycloalkyl;

Is aryl, substituted aryl, arylfluorenyl, C! -Ctfluorenyl or c ₃ -c ₇ cyclofluorenyl;

X is NR ₇ , 0 or S;

R ₃ is aryl, substituted aryl, arylfluorenyl, -C4 alkylaryl or C ₃ -C ₇ cycloalkyl;

F is hydroxyl or ^-^ 垸;

5 is hydroxyl, C _r C ₆ fluorenyl, -C (0) R ₈ ,-(CH ₂ ) _m R ₈ , -CH ₂ CH = CHR ₈ , -C (0) OR ₈ or -S (0) ₂ R ₈ , m is 0, 1, 2 or 3;

R ₆ is hydroxy or ^-^-;

R ₇ is hydrogen, hydroxyl or -fluorenyl;

2. The piperazine triazine compound or a pharmaceutically acceptable salt thereof according to claim 1, wherein X is NR ₇ and R ₇ is hydrogen.

3. The piperazine triazine compound or a pharmaceutically acceptable salt thereof according to claim 1, wherein X is NR ₇ and R ₇ is a hydroxyl group.

4. The piperazine triazine compound or a pharmaceutically acceptable salt thereof according to claim 1, wherein X is NR ₇ and R ₇ is a dC ₆ alkyl group.

5. The piperazine triazine compound or a pharmaceutically acceptable salt thereof according to claim 1, wherein X is 0.

6. The piperazine triazine compound or a pharmaceutically acceptable salt thereof according to claim 1, wherein

乂 is S.

7. A triazine compound represented by formula (II),

(II)

Wherein, for the 1, dC ₄ alkyl group, an aryl group, a substituted aryl group, an aryl group embankment, dC ₄ embankment acyl, aroyl, or C ₃ -C ₇ cycloalkyl group embankment;

R ₂ is aryl, substituted aryl, arylfluorenyl, fluorenylaryl or C ₃ -C ₇ cyclofluorenyl;

X is NR ₇ , 0 or S;

Aryl, substituted aryl, arylfluorenyl, fluorenyl or C ₃ -C ₇ cycloalkyl;

Y is hydrogen, halogen or -C4 alkylamine.

8; a triazine compound represented by formula (III),

(III)

In the formula, R is H, dC ₄ fluorenyl, aryl, substituted aryl, aryl fluorenyl, d-alkanoyl, aroyl or C ₃ -C ₇ cycloalkyl;

Is benzene, aryl, substituted aryl, arylfluorenyl, C fluorenyl or C ₃ -C ₇ cycloalkyl;

Y is hydrogen, halogen or amino-fluorenyl;

Z is hydrogen, halogen or aminoC fluorenyl.

Preparation method of piperazinetriazine compound represented by formula (I) or pharmaceutically acceptable salt thereof

(I)

Where

R is H, CC ₄ alkyl, aryl, substituted aryl, arylfluorenyl, cc ₄ alkanoyl, aroyl, or C ₃ -C ₇ ring

'j¾ 基;

R ₂ is aryl, substituted aryl, arylfluorenyl, dc ₄ fluorenyl or c ₃ -c ₇ cyclofluorenyl;

X is NR ₇ , 0 or S;

R ₃ is aryl, substituted aryl, arylfluorenyl, C _r C ₄ aryl or C ₃ -C ₇ cycloalkyl;

R4 is hydroxyl or -0; fluorenyl;

R ₅ is hydroxyl, C, -C ₆ fluorenyl, -C (0) R ₈ ,-(CH ₂ ) _m R ₈ , -CH ₂ CH = CHR ₈ , -C (0) OR ₈ or-S (0 ) ₂ R ₈ , m is 0, 1, 2 or 3;

R ₆ is hydroxyl or -C ₆ fluorenyl;

R ₇ is hydrogen, hydroxy or Cr alkyl;

Hydrogen, hydroxy, aryl, heteroaryl ring, heterocyclyl or C2-C6 alkenyl hydrocarbon

The method mainly includes the following steps:

(1) Formula (IV)

Compound

(IV)

In the formula, M is hydrogen, halogen or amino CC ₄ alkyl, and Y and Z are the same as defined in the general formulae (1), (11), and (III), and under the conditions of inert solvent and basic conditions, it is 20-10 (TC Reaction with a substituted amine to obtain a compound represented by the general formula (III):

(III)

In the formula, R ,, R ₂ , Y, and Z are the same as defined in the general formulae (1) and (II);

(2) The compound represented by the general formula (III) is reacted with an aromatic amine, alcohol or alcohol at 20-100 ° C under an inert solvent and basic conditions to obtain a compound represented by the general formula (Π):

In the formula, R,, R ₂ , R ₃ , X, Y are the same as defined in the general formula (I);

(3) reacting a compound represented by the general formula (II) with a substituted piperazine under -40-40 ° C and basic conditions to obtain a compound represented by the formula (I);

(4) If necessary, a salt formation reaction is performed.

10. The method according to claim 9, wherein the inert solvent used in step (2) and step (3) is selected from the group consisting of THF, Et ₂ 0, DMF, ethylene glycol dimethyl ether, ethylene glycol diethyl ether, and Dioxane, the base used is selected from organic bases including pyridine, triethylamine, 4-dimethylaminopyridine (DMAP), diisopropylethylamine and sodium carbonate, potassium carbonate, sodium hydroxide, hydrogen Inorganic base of potassium oxide. '·

1 1. The preparation method according to claim 9, wherein the polar organic solvent used in step (3) is selected from the group consisting of ethanol, methanol, ethyl acetate and tetrahydrofuran, and the base used is selected from the group consisting of pyridine, triethylamine, 4 -Organic bases of dimethylaminopyridine (DMAP), diisopropylethylamine and inorganic bases including sodium carbonate, potassium carbonate, sodium hydroxide, potassium hydroxide.

12. The preparation method according to claim 9, wherein the products obtained in steps (1), (2) and (3) can be purified separately to obtain pure products.

13. The method according to claim 12, wherein the purification method comprises column chromatography or recrystallization.

14. A pharmaceutical composition for the prevention and / or treatment of inflammatory diseases, comprising a therapeutically effective amount of a piperazinetriazine compound of the formula (I) or a pharmaceutically acceptable salt thereof and at least one pharmaceutically acceptable carrier.

15. The pharmaceutical composition for preventing and / or treating inflammatory diseases according to claim 14, wherein the pharmaceutically acceptable carrier comprises an ion exchanger, alumina, aluminum stearate, lecithin, serum protein, buffer Substances such as phosphate, glycerol, sorbic acid, potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids, water, salts or electrolytes, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, Magnesium trisilicate, polyvinylpyrrolidone, cellulose materials, polyethylene glycol, sodium carboxymethyl cellulose, polyacrylates, beeswax and lanolin. '