CN100432071C - Substituted 1H-indol-2-one compound and its preparation method and use - Google Patents

Abstract

The present invention relates to a novel substituted 1H-indole-2-ketone compound and an application thereof. The structure of the compound is disclosed in the right formula. A pharmacological experiment indicates that the compound has certain pharmacology performance and especially has the inhibitory action on the tyrosine kinase of an epidermal growth factor receptor and the associated hyperplasia of various tumor cells thereof. The present invention also relates to a preparation method of the compound.

Description

Substituted 1H-indol-2-one compounds and their preparation methods and uses

technical field

The present invention relates to a class of substituted 1H-indol-2-one compounds, in particular to a class of substituted 1H-indol-2-one compounds that have an inhibitory effect on epidermal growth factor tyrosine kinase, and the present invention also relates to the Method for the preparation of compounds.

Background technique

The role of protein tyrosine kinases (PTKs) is to transfer the phosphate group of ATP to specific tyrosine residues in functional proteins, and this tyrosine phosphorylation process can regulate cell growth, proliferation and differentiation. The number of protein tyrosine kinases is estimated to exceed 1000, of which 100 have been sequenced. Protein tyrosine kinase plays a very important role in the process of tumorigenesis. It is known that more than half of the proto-oncogenes express proteins with PTK activity. The important role of protein tyrosine kinase in cancer cell proliferation makes it an extremely important Attractive target for cancer therapy.

The most widely studied protein tyrosine kinase is the epidermal growth factor receptor (EGFR). EGFR is a 170KDa transmembrane glycoprotein, and its structure includes an extramembrane ligand-binding domain, a transmembrane domain, and an intracellular domain (with PTK activity) and a C-terminal domain. The epidermal growth factor receptor family includes four members: EGFR (ErbB-1), ErbB-2 (HER2), ErbB-3 (HER3), ErbB-4 (HER4). Cancer clinical studies have shown that the receptor and its ligands are closely related to many tumors, and EGFR is overexpressed in 60% of cancers. This abnormality manifests in many forms, but each causes excess tyrosine phosphorylation signaling into the cell. In cellular systems, EGF or other ErbB family ligands often cause uncontrollable cell proliferation, thereby transforming cells from normal to typically abnormal forms. Binding of extramembrane ligands to EGFR causes EGFR receptors to form dimers (homodimers or heterodimers), activate tyrosine kinases, and transmit proliferative signals to cells. Using drugs to act on EGFR can stop the proliferation signal mediated by it, thereby preventing the excessive reproduction of cancer cells. There are two classes of drugs that can do this: monoclonal antibodies and small molecules. The mode of action of monoclonal antibody drugs is to occupy the extramembrane ligands of EGFR, so that endogenous ligands such as EGF cannot bind to EGFR, thereby preventing the signal from passing into cells. Small-molecule drugs block cell proliferation signals by blocking the catalytic domain of EGFR intracellular tyrosine kinase and preventing tyrosine phosphorylation.

Small molecule EGFR inhibitors include natural inhibitors and organic synthetic compounds. Natural inhibitors have good inhibitory activity on EGFR, but due to reasons such as cytotoxicity, enzyme selectivity (especially kinase selectivity) and mechanism of action, no natural inhibitor nucleus with promising drug development has been found yet. The research on natural inhibitors is mainly to find EGFR inhibitors by modifying the structure of natural products (Alexander J. Bridges, Chem. Rev. 2001, 101, 2541-2571).

Organic synthesis small molecule inhibitors can be divided into reversible inhibitors and irreversible inhibitors according to the mode of action with EGFR, and can be divided into 4-anilino (hetero)cyclopyrimidines and 4-anilino-3- Cyanoquinolines. 4-Anilino (hetero)cyclopyrimidines are the most widely studied class of EGFR inhibitors, including 4-anilinoquinazolines, 4-anilinopyridopyrimidines and 4-anilinopyrrolopyrimidines, etc. , of which 4-anilinoquinazolines are the most representative.

Zeneca company discloses a kind of inhibition EGFR IC ₅₀ value reaches 20-40nM, and has better selectivity 4-anilinoquinazoline compound (European Patent Publication No. 0520722A, 1992); Fry has reported a kind of inhibition The IC ₅₀ value of EGFR is 29pM, and it is a specific EGFR inhibitor compound. This compound is a reversible ATP-binding domain competitive inhibitor. It has shown good enzyme selectivity in cell level experiments and inhibited the process of EGFR autophosphorylation The _IC50 value of 14nM, but has the disadvantage of poor solubility. (Fry, Science, 1994, 265:1093-1095).

是于2003年5月获得FDA批准上市的EGFR抑制剂，属4-苯氨基喹唑啉类化合物，是可逆的ATP结合域结合的竞争抑制剂，它在酶水平的EGFR抑制活性IC₅₀值为23nM，对EGF刺激繁殖的口腔上皮鳞癌细胞KB的抑制作用的IC₅₀值为80nM，具备高度的EGFR酶专一性，对其他激酶的抑制能力很低。研究表明Iressa有很好的生物利用度和极低的毒性，对非小细胞肺癌和乳腺癌也有明显的治疗作用。但部分研究显示Iressa在联合化疗时并不能提高生存率或改善症状；另外，在日本的一年临床实践发现有小部分患者出现了严重的毒副反应，如间质性肺炎(ILD)，并有部分病人死亡。Iressa主要毒副反应包括：腹泻，恶心，呕吐，皮疹和痤疮等，另外可能使服用抗凝药物华法令的患者出现出血症状。

It is an EGFR inhibitor that was approved by the FDA in May 2003. It belongs to 4-anilinoquinazoline compounds and is a reversible competitive inhibitor of ATP-binding domain binding. Its EGFR inhibitory activity at the enzyme level has an IC ₅₀ value of 23nM, the IC ₅₀ value of the inhibitory effect on EGF-stimulated oral squamous cell carcinoma KB is 80nM, which has a high degree of EGFR enzyme specificity and low inhibitory ability to other kinases. Studies have shown that Iressa has good bioavailability and extremely low toxicity, and it also has obvious therapeutic effects on non-small cell lung cancer and breast cancer. However, some studies have shown that Iressa can not improve the survival rate or improve symptoms when combined with chemotherapy; in addition, a year of clinical practice in Japan found that a small number of patients had severe side effects, such as interstitial pneumonia (ILD), and Some patients died. The main toxic and side effects of Iressa include: diarrhea, nausea, vomiting, skin rash and acne, etc. In addition, it may cause bleeding symptoms in patients taking the anticoagulant drug warfarin.

According to reports, 6,7-dimethoxy-4-anilino-3-cyanoquinoline has an inhibitory activity _IC50 value of 190nM for EGFR enzyme levels; the formyl group is substituted on the 3-position of the mother nucleus, and its inhibition The activity reaches the μM level, but this class of inhibitors is slightly inferior in enzyme selectivity, and they also have c-SRC and MEK inhibitory activity. (Wang YD, Miller K., Boschelli DH et al., Bioorg. Med. Chem. Lett. 2000, 10: 2477-2480; Boschelli DH, Yang YD, Ye F. et al., J. Med. Chem. 2001, 44: 822 -846) In addition, 2,3-dihydro-2-thione-indole-3-alkylcarboxylic acid compounds and 2,2'-dithiobisindole-3-alkylcarboxylic acid compounds have been reported as EGFR (Zhang N., Wu B., Powell D., Wissner A. et al., Bioorg. Med. Chem. Lett. 2000, 10: 2825-2829).

The above-mentioned inhibitors are all reversible inhibitors that compete with ATP for binding. Although they are reversible inhibition, their dissociation speed from the enzyme is very slow. The researchers believe that the binding of these "quasi-irreversible inhibitors" to the receptor can cause the so-called "hydrophobic collapse" of the receptor, which leads to the closure of the kinase domain of the receptor and prevents the protein phosphorylation process. The concentration of ATP in cells is in mM level, so in the early clinical stage, the dosage of reversible inhibitors is very large, the purpose is to maintain the required blood concentration, and the longest time for blocking the combination of enzyme and ATP can reach 4 hours, and less soluble drug molecules are more effective than easily soluble drug molecules. In view of this, it is necessary to find irreversible inhibitors of EGFR.

The EGFR irreversible inhibitors that have entered the clinical trial stage include 4-anilinoquinazolines and 4-anilino 3-cyanoquinolines (Alexander J. Bridges, Chem.Rev.2001, 101, 2541 -2571).

There are still many problems in the development of EGFR inhibitors, such as the unknown toxicity of 4-anilinoquinazoline compounds caused by the failure of Iressa's clinical trials in Japan, and the problem of enzyme selectivity. To solve these problems and develop new EGFR inhibitors is a hot research topic in the world of medicine.

Contents of the invention

The purpose of the present invention is to provide a class of novel substituted 1H-indol-2-one compounds that can inhibit the expression of epidermal growth factor to achieve antitumor drug effect.

Another object of the present invention is to provide a process for the preparation of the compounds of the present invention.

Another object of the present invention is to provide the application of the compound of the present invention as an inhibitor of epidermal growth factor tyrosine kinase in the preparation of antitumor drugs.

The compound of the present invention is a substituted 1H-indol-2-one compound having the structure of the following general formula (I):

in:

_R is selected from hydrogen, C _1-5 alkyl, C _2-5 alkenyl, C _3-10 cycloalkyl, hydroxyl, C _1-5 alkoxy, acetyl, carboxyl, amido, thioamide group, sulfonyl group and trihalomethylsulfonyl group;

R ₂ , R ₃ , R ₄ , and R ₅ are each independently selected from hydrogen, C _1-5 alkyl, C _1-4 alkoxy, aryl, aryloxy, C _1-5 alkylaryl, C _1-5 alkyl aryloxy, C _3-10 cycloalkyl, mercapto, C _1-5 alkyl mercapto, aryl mercapto, sulfinyl, halogen, trihalomethyl, amino, cyano, nitro, carboxyl, Oxycarboxyl, amido, thioamido, sulfonamide, trihalomethylsulfonyl, N-sulfinamide, S-sulfinamide, trihalomethylsulfinyl, O-carbamoyl, O-thiocarbamoyl and -NR ₁₁ R ₁₂ , wherein R ₃ and R ₄ , R ₄ and R ₅ can combine to form a six-membered aromatic ring, methylenedioxy or ethylenedioxy;

R ₄ and R ₅ can also be selected from the following groups simultaneously or separately:

Wherein _R is selected from hydrogen; C _1-5 alkyl; C _1-5 alkoxy; aryl; aryloxy; C _1-5 alkylaryl; C _1-5 alkylaryloxy; trihalogen Methyl; pyridyl; pyrimidinyl; thienyl; each of the above groups may be substituted by: trifluoromethoxy; fluorine; chlorine; bromine; iodine; C _1-3 -alkoxy or substituted C _1-3 -alkoxy, wherein substituted C _1-3 -alkoxy means that the 2- or 3-position is replaced by amino, C _1-3 -alkylamino, 2-(C _1-3 -alk base)amino, phenyl-(C _1-3 -alkyl)amino, N-(C _1-3 -alkyl)-phenyl-(C _1-3 -alkyl)amino, pyrrolidinyl Or piperidinyl substitution; phenyl-C _1-3 -alkylamino-C _1-3 -alkyl, the substituent can also be trifluoromethyl, fluorine, chlorine, bromine, iodine, C _1-5 -Alkyl or C _1-3 -alkoxy can mono-substitute or di-substitute the phenyl nucleus. When di-substituted, the substituents can be the same or different;

R ₆ is selected from hydrogen, C _3-10 cycloalkyl, C _1-5 alkyl, halogen;

R and R are _each selected from hydrogen, C _1-5 alkyl _, C _1-4 alkoxy, aryloxy, C _1-5 alkylaryl, C _1-5 alkylaryloxy, trihalogen C _1-5 alkyl, C _3-10 cycloalkyl, mercapto, C _1-5 alkyl mercapto, aryl mercapto, sulfinyl, halogen, trihalomethyl, amino, cyano, nitro, carboxyl, oxygen Carboxyl, amide, thioamido, sulfonamide, trihalomethylsulfonyl, N-sulfinamide, S-sulfinamide, trihalomethylsulfinyl, O-carbamoyl, O -thiocarbamoyl and -NR ₁₁ R ₁₂ ;

_R is selected from hydrogen, C _1-5 alkyl, C _1-5 alkoxy, aryl, aryloxy, C _1-5 alkylaryl, C _1-5 alkylaryloxy, halogen, tri Halomethyl, mercapto, hydroxyl, thio C _1-5 alkyl and -NR ₁₁ R ₁₂ ;

R ₁₀ is selected from hydrogen, C _3-10 cycloalkyl, C _1-5 alkyl, C _2-5 alkenyl, aryl, heteroaryl, hydroxyl and heteroalicyclic;

R ₁₁ and R ₁₂ are independently selected from hydrogen, C _1-5 alkyl, C _2-5 alkenyl, C _3-10 cycloalkyl, acetyl, sulfonyl, trifluoromethanesulfonyl.

A preferred compound of the compound of the present invention is a 1,4-disubstituted benzene compound with a structure of general formula (I), wherein _R is selected from C _1-5 alkyl, halogen, sulfonamide, amino, cyano, nitrate Group, carboxyl and -NR ₁₁ R ₁₂ ; R ₁ , R ₂ , R ₃ , R ₅ , R ₆ , R ₇ , R ₈ , R ₉ , R ₁₀ , R ₁₁ , R ₁₂ are as defined in the aforementioned compounds.

Another preferred compound of the present invention is a 1,4-disubstituted benzene compound with a structure of general formula (I), wherein R ₄ and R ₅ are simultaneously or respectively selected from the following groups:

The definitions of R ₁ , R ₂ , R ₃ , R ₄ , R ₇ , R ₈ , R ₉ , R ₁₀ , R ₁₁ , R ₁₂ and R ₁₃ are the same as those for the aforementioned compounds.

In the present invention, aryl in aryl or aryl-containing groups refers to phenyl, naphthyl and biphenyl; heteroaryl refers to single ring and condensed ring (i.e. ring) groups having 1 to 3 atoms selected from nitrogen, oxygen and sulfur on the ring and additionally having a fully connected π-electron system, for example pyrrole, furan, thiophene, imidazole, thiazole, pyrazole, pyridine, Pyrimidine, quinoline, isoquinoline, purine and carbazole, etc.; heteroalicyclic group refers to a single ring or a condensed ring (C _3-15 ) having an atom selected from nitrogen, oxygen and sulfur atoms on the ring, said Rings may also contain 1 to 7 double bonds, however, said rings do not have a fully connected π-electron system, such as piperidine, tetrahydrofuran, tetrahydropyrrole, tetrahydrothiophene, cyclohexanopyrrole, and the like.

Preferred compounds of the present invention include: (E)-3-[(3-ethoxycarbonylmethyl-2-ethoxycarbonyl-5-methyl-1H-pyrrol-4-yl)methylene]-1H -indol-2-one; (E)-5-bromo-3-[(3-ethoxycarbonylmethyl-2-ethoxycarbonyl-5-methyl-1H-pyrrol-4-yl)ylidene Methyl]-1H-indol-2-one; (E)-3-[(3-methoxycarbonylmethyl-2-methoxycarbonyl-5-methyl-1H-pyrrol-4-yl) Methylene]-(1H)-indol-2-one; (E)-5-bromo-3-[(3-methoxycarbonylmethyl-2-methoxycarbonyl-5-methyl-1H -pyrrol-4-yl)methylene]-(1H)-indol-2-one; (E)3-[(3,5 dimethyl-2-ethoxycarbonyl-1H-pyrrole-4- Base) methylene]-(1H)-indol-2-one; (E)-5-bromo-3-[(3,5 dimethyl-2-ethoxycarbonyl-1H-pyrrole-4- Base) methylene]-(1H)-indol-2-one; (E)-5-nitro-3-[(3,5 dimethyl-2-ethoxycarbonyl-1H-pyrrole-4 -yl)methylene]-(1H)-indol-2-one; (E)3-[(3,5 dimethyl-2-methoxycarbonyl-1H-pyrrol-4-yl)methylene Base]-(1H)-indol-2-one; (E)-5-nitro-3-[(3,5 dimethyl-2-methoxycarbonyl-1H-pyrrol-4-yl)ylidene Methyl]-(1H)-indol-2-one; (E)-5-nitro-3-[(3-ethoxycarbonylmethyl-2-ethoxycarbonyl-5-methyl-1H -pyrrol-4-yl)methylene]-(1H)-indol-2-one; (E)-5-carboxy-3-[(3-ethoxycarbonylmethyl-2-ethoxycarbonyl -5-Methyl-1H-pyrrol-4-yl)methylene]-(1H)-indol-2-one; (E)-5-aminosulfonyl-3-[(3-ethoxycarbonyl Methyl-2-ethoxycarbonyl-5-methyl-1H-pyrrol-4-yl)methylene]-(1H)-indol-2-one; (E)-5-methylaminosulfonyl -3-[(3-ethoxycarbonylmethyl-2-ethoxycarbonyl-5-methyl-1H-pyrrol-4-yl)methylene]-(1H)-indol-2-one; (E)-5-(N-morpholinyl)aminosulfonyl-3-[(3-ethoxycarbonylmethyl-2-ethoxycarbonyl-5-methyl-1H-pyrrol-4-yl) Methylene]-(1H)-indol-2-one; (E)-5-(3-chlorophenyl)aminosulfonyl-3-[(3-ethoxycarbonylmethyl-2-ethoxy Cylcarbonyl-5-methyl-1H-pyrrol-4-yl)methylene]-(1H)-indol-2-one; (E)-5-(3-fluorophenyl)aminosulfonyl-3 -[(3-ethoxycarbonylmethyl-2-ethoxycarbonyl-5-methyl-1H-pyrrol-4-yl)methylene]-(1H)-indol-2-one; (E )-5-dimethylaminosulfonyl-3-[(3-ethoxycarbonylmethyl-2-ethoxycarbonyl-5-methyl-1H-pyrrol-4-yl)methylene]-( 1H)-indol-2-one; (E)-5-(3-fluoro-4-chlorophenyl)aminosulfonyl-3-[(3-ethoxycarbonylmethyl-2-ethoxycarbonyl -5-Methyl-1H-pyrrol-4-yl)methylene]-(1H)-indol-2-one; (E)-5-(hexahydropyridyl)aminosulfonyl-3-[( 3-Ethoxycarbonylmethyl-2-ethoxycarbonyl-5-methyl-1H-pyrrol-4-yl)methylene]-(1H)-indol-2-one; (E)-5 -(3-bromophenyl)aminosulfonyl-3-[(3-ethoxycarbonylmethyl-2-ethoxycarbonyl-5-methyl-1H-pyrrol-4-yl)methylene]- (1H)-Indol-2-one.

The preparation of the compounds of the present invention is carried out through the following steps:

The compound of the present invention (compound I) is prepared by condensation of substituted 1H-indol-2-one (compound II) and substituted 3-formyl 1H-pyrrole (compound III): under the catalysis of base, equimolar compound II and Compound III undergoes a condensation reaction in an organic solvent. According to the reaction of the specific compound, the reaction temperature is 65°C to 90°C; the reaction time is 3 to 6 hours. Column chromatography separation, etc. The final product (I) is detected and proved by nuclear magnetic resonance or mass spectrometry.

Among the present invention, an organic solvent refers to a protic solvent or an aprotic solvent, wherein the protic solvent is water or alcohols (such as ethanol, methanol, n-butanol); the aprotic solvent is a polar or nonpolar aprotic solvent, and the aprotic solvent The feature is that it does not contain acidic hydrogen, that is, it does not contain hydrogen that can be combined with the solute. The preferred non-polar aprotic solvent is pentane, hexane, benzene, toluene, methylene chloride or carbon tetrachloride. The preferred polar nature The sub-inert solvent is chloroform, tetrahydrofuran, dimethylsulfoxide or dimethylformamide. Alkali catalysts refer to organic nitrogen bases and inorganic bases, wherein organic nitrogen bases can be triethylamine, pyridine, hexahydropyridine, trimethylamine, aniline, diisopropylamine, 1,8-diazabicyclo[5.4.1 ] Undec-7-ene, pyrrolidine, piperidine etc., inorganic base can adopt ammonia, alkali metal hydroxide or alkaline earth metal hydroxide (such as lithium hydroxide, sodium hydroxide, potassium hydroxide, calcium hydroxide, Magnesium hydroxide, barium hydroxide) phosphate, carbonate, bicarbonate, bisulfate, etc. When the solvent is a protic solvent, the base catalyst is an alkali metal hydroxide or an alkaline earth metal hydroxide.

In the present invention, the synthesis method of compound II refers to the patent method (WO9961442);

In the present invention, the synthetic method of compound III refers to literature method (Alan R.Battersby., Eric Hunt et al., J.Chem.Soc.Perk.I 1976:1008-1035; Woodward, R.B.J.Am.Chem.Soc.1960 , 82, 3800-3892.).

Beneficial effect

The present invention designs and synthesizes a new class of substituted 1H-indol-2-one compounds, which are effective against human autosomal dominant polycystic kidney disease (ADPKD) cyst lining epithelial cells, A-549 human lung adenocarcinoma cells, A-431 human skin squamous cell carcinoma cells and MDA-MB-468 human breast cancer cells have inhibitory effects, and can be used to prepare medicines for treating diseases caused by excessive or abnormal cell proliferation. The compound of the invention has a relatively simple structure and is easy to prepare.

Detailed ways

The present invention will be further described below in conjunction with specific examples, but the present invention is not limited.

In the following preparation examples, the melting point is measured by the X-4 melting point apparatus produced by Shanghai Precision Scientific Instrument Co., Ltd., and the temperature is uncorrected; the nuclear magnetic resonance is measured by the Bruker AMX-400 type and INVOA-600 type nuclear magnetic resonance apparatus, and TMS is the internal Standard, chemical shift unit is ppm; mass spectrometry is measured by MAT-711 and MAT-95 mass spectrometer; silica gel 200-300 mesh for column chromatography, produced by Qingdao Ocean Chemical Factory; TLC silica gel plate is HSGF- 254 prefabricated plates for thin layer chromatography; the boiling range of petroleum ether is 60-90°C; the color is developed by ultraviolet lamp and iodine cylinder. If the operation method is not specified in the preparation examples, the concentration refers to evaporating the solvent in the solution of the prepared compound with a rotary evaporator; the drying refers to drying the prepared compound at 80° C. with a DHG-9240A constant temperature drying oven.

Example 1 (E)-3-[(3-ethoxycarbonylmethyl-2-ethoxycarbonyl-5-methyl-1H-pyrrol-4-yl)methylene]-1H-indole- Preparation of 2-keto (compound I-1)

Under an ice bath at 0-10°C, add aqueous sodium nitrite solution (sodium nitrite 29g, water 60ml) into a solution of 2-carbonyl-1,3-diethyl malonate (80g) in acetic acid (140ml) and stir 5 hours. A solution of benzyl acetoacetate (76g) in acetic acid (140ml) was added to the reaction solution, zinc powder (46g) and anhydrous sodium acetate (40g) were added, and stirring was continued for 15 minutes. The ice bath was removed, heated to 80°C with an oil bath, and stirred for 4 hours. The reaction solution was poured into crushed ice (1Kg), left overnight, filtered, and concentrated to obtain 60 g of a light yellow solid. Recrystallized from ethanol to obtain 51 g of white solid 4-benzyloxycarbonyl-3-ethoxycarbonylmethyl-2-ethoxycarbonyl-5-methyl-1H-pyrrole with a yield of 33%.

Dissolve 4-benzyloxycarbonyl-3-ethoxycarbonylmethyl-2-ethoxycarbonyl-5-methyl-1H-pyrrole (51g, 136mmol) in tetrahydrofuran (50ml), add 10% palladium carbon (10g), room temperature, feed hydrogen, after stirring for 1 hour, filter off the palladium carbon, wash the filter cake with tetrahydrofuran (50ml), concentrate and dry the filtrate to obtain 36g white solid 3-ethoxycarbonylmethyl-2-ethoxy Carbonyl-5-methyl-1H-pyrrole-4-carboxylic acid, yield 95%.

3-Ethoxycarbonylmethyl-2-ethoxycarbonyl-5-methyl-1H-pyrrole-4-carboxylic acid (36g) was dissolved in 1,2-dichloroethane (200ml), oil bath Heat to 50°C. Add aqueous sodium bicarbonate solution (sodium bicarbonate 40g: water 200ml), stir, and add iodine aqueous solution (iodine 240g: water 200ml) containing sodium iodide (20g) dropwise after bubbling. Continue to heat and stir for half an hour, and add sodium bisulfite to remove excess iodine after the TLC detection method is followed to determine that the reaction is complete. The organic layer was separated, dried and concentrated to give 42 g of crude product. Recrystallization from 1,2-dichloroethane afforded 40 g of pure 3-ethoxycarbonylmethyl-2-ethoxycarbonyl-4-iodo-5-methyl-1H-pyrrole as a white solid, yield 91%.

3-Ethoxycarbonylmethyl-2-ethoxycarbonyl-4-iodo-5-methyl-1H-pyrrole (40 g) was dissolved in methanol (200 ml), and the oil bath was heated to reflux. After adding an aqueous solution of sodium iodide (25 g of sodium iodide: 30 ml of water), concentrated hydrochloric acid (37%) (10 ml) was added, and after 15 minutes, sodium bisulfite (500 mg) was added to remove excess iodine. The methanol in the reaction solution was distilled off (75°C), extracted with 1,2-dichloroethane (100ml), the organic layer was concentrated and dried to obtain 29g of 3-ethoxycarbonylmethyl-2-ethoxy Carbonyl-5-methyl-1H-pyrrole, white solid, yield 87%.

Phosphorus oxychloride (15ml) was added dropwise to N,N-dimethylformamide (15ml) at 0-5°C. After stirring for 15 minutes, a solution of 3-ethoxycarbonylmethyl-2-ethoxycarbonyl-5-methyl-1H-pyrrole (29g) in 1,2-dichloroethane (100ml) was slowly added dropwise. Stirring was continued for 15 minutes under an ice bath at 0-5°C. Remove the ice bath and heat to 50°C with an oil bath. After half an hour, add an aqueous solution of sodium acetate (10 g of anhydrous sodium acetate: 50 ml of water). After half an hour, the reaction solution was cooled, and the organic layer was separated with a separatory funnel and concentrated, and separated on a silica gel column with an eluent of ethyl acetate:petroleum ether=1:3 to obtain 26g of 3-ethoxycarbonylmethyl -2-Ethoxycarbonyl-4-formyl-5-methyl-1H-pyrrole, white solid, yield 80%.

3-Ethoxycarbonylmethyl-2-ethoxycarbonyl-4-formyl-5-methyl-1H-pyrrole (1g) and 1H-indol-2-one (0.5g) in ethanol (20ml ), heated the oil bath to 80°C, and added hexahydropyridine (0.1g) dropwise. After reacting for 3 hours, the reaction solution was concentrated, dissolved in 1,2-dichloroethane (10ml), dried and washed with 20ml of water and 20ml of 2N hydrochloric acid, concentrated organic layer, and eluted with ethyl acetate:petroleum ether=1:4 The solution was purified on a silica gel column to obtain 1.2 g of (E)-3-[(3-ethoxycarbonylmethyl-2-ethoxycarbonyl-5-methyl-1H-pyrrol-4-yl)methylene ]-1H-indol-2-one, yellow solid, yield 83%.

H ¹ NMR (CDCl ₃ ) 9.20 (s, 1H, NH) 9.90 (s, 1H, NH) 7.82 (s, 1H, vinyl-H) 7.20 (t, 2H, Ph-H) 7.09 (t, 1H, Ph -H) 6.90 (m, 2H, Ph-H) 4.33 (q, 2H, CO ₂ CH ₂ CH ₃ ) 4.08 (q, 2H, CO ₂ CH ₂ CH ₃ ) 3.80 (s, 3H, CH ₂ CO ₂ CH ₂ CH ₃ ) 2.21 (s, 3H, CH ₃ ) 1.36 (t, 3H, CO ₂ CH ₂ CH ₃ ) 1.20 (t, 3H, CO ₂ CH ₂ CH ₃ ).

Example 2 (E)-5-bromo-3-[(3-ethoxycarbonylmethyl-2-ethoxycarbonyl-5-methyl-1H-pyrrol-4-yl)methylene]-1H - Preparation of indol-2-one (compound 1-2):

1H-Indol-2-one (1.3g) in acetonitrile (20ml) was cooled to -10°C and N-bromosuccinimide (2g) was added slowly with stirring. The reaction was stirred at -10°C for 1 hour and at 0°C for 2 hours. The precipitate was collected, washed with 20 ml of water and dried to give 1.9 g of 5-bromo-1H-indol-2-one, yield 90%.

3-Ethoxycarbonylmethyl-2-ethoxycarbonyl-4-formyl-5-methyl-1H-pyrrole (0.95g, 3.6mmol) and 5-bromo-(1H)-indole-2 - Ketone (0.8g, 3.6mmol) was mixed and stirred in 20ml ethanol, heated to 80°C in an oil bath, hexahydropyridine (0.1g) was added dropwise, and the reaction solution was concentrated after 2.5 hours of reaction, and 1,2-dichloroethane was added (10ml), dried and washed with 20ml of water and 20ml of 2N hydrochloric acid, the organic layer was concentrated, and purified on a silica gel column with an eluent of ethyl acetate:petroleum ether=1:4 ( _Rf =0.6) to obtain 1.56g of a yellow solid ( E)-5-bromo-3-[(3-ethoxycarbonylmethyl-2-ethoxycarbonyl-5-methyl-1H-pyrrol-4-yl)methylene]-(1H)-ind Indol-2-one, 90% yield.

H ¹ NMR (DMSO-d6) 7.48 (s, 1H, vinyl-H) 7.38 (d, 2H, Ph-H) 6.96 (s, 1H, Ph-H) 6.80 (q, 2H, Ph-H) 4.23 ( q, 2H, CO ₂ CH ₂ CH ₃ ) 3.98 (q, 2H, CO ₂ CH ₂ CH ₃ ) 3.75 (s, CH ₂ CO ₂ CH ₂ CH ₃ ) 2.09 (s, 3H, CH ₃ ) 1.36( _t , 3H _, CO2CH2CH3 ) 1.20(t _, 3H _{, CO2CH2CH3 )} . MS m/e 460 (M ⁺ ).

Example 3 (E)-3-[(3-methoxycarbonylmethyl-2-methoxycarbonyl-5-methyl-1H-pyrrol-4-yl)methylene]-(1H)-ind Preparation of indol-2-one (compound 1-3):

Under ice bath at 0-10°C, aqueous sodium nitrite solution (19.8 g of sodium nitrite: 100 ml of water) was added dropwise to acetic acid (150 ml) of dimethyl 2-carbonyl-1,3-malonate (50 g, 287 mmol) ) solution, the ice bath was removed, and stirred at room temperature for 5 hours, the solution was brownish red. In an ice bath, benzyl acetoacetate (59g, 287mmol) was added to the reaction solution, then acetic acid (50ml) was added, zinc powder (25g) and anhydrous sodium acetate (20g) were added, and stirring was continued for 15 minutes, then removed Ice bath, heated to 80°C with an oil bath, all the solids were dissolved, and after stirring for 4 hours, the reaction solution was poured into ice water solution, left overnight, filtered, and the filtrate was concentrated to obtain 35 g of light yellow solid. After recrystallization from ethanol, 34 g of 4-benzyloxycarbonyl-3-methoxycarbonylmethyl-2-methoxycarbonyl-5-methyl-1H-pyrrole were obtained, yield 33%. Mp148°C. H ¹ NMR (CDCl ₃ ): 9.0 (broad, 1-H), 7.3-7.4 (m, 5H, Ph-H), 5.25 (s, 2H, CH ₂ Ph), 4.2 (s, 2H, CH _{2 CO2Me} ), 2.5 ( _s , 3H _, CH3 ) , 1.35 ( s , 3H, _CO2CH3 ), 1.2 (s, 3H, _{CO2CH3 )} .

30g of 4-benzyloxycarbonyl-3-methoxycarbonylmethyl-2-methoxycarbonyl-5-methyl-1H-pyrrole (87mmol) was dissolved in tetrahydrofuran (50ml), and 10% palladium carbon ( 5g), room temperature, hydrogen flow, after stirring for 1 hour, filter off palladium carbon, wash the filter cake with tetrahydrofuran (50ml), the filtrate is concentrated and dried to obtain 22g of 3-methoxycarbonylmethyl-2-methoxycarbonyl- 5-Methyl-1H-pyrrole-4-carboxylic acid, 95% yield. Mp243°C. H ¹ NMR (CDCl ₃ ): 9.6 (br, 1-H, COOH ), 3.80 (s, 2H, CH ₂ CO ₂ Me), 2.3 (s, 3H, CH ₃ ), 1.3 (s, 3H, _CO2CH3 ) , 1.2 (s _, 3H , _CO2CH3 ) _.

Dissolve 20 g of 3-methoxycarbonylmethyl-2-methoxycarbonyl-5-methyl-1H-pyrrole-4-carboxylic acid (74 mmol) in 1,2-dichloroethane (100 ml), oil Heat the bath to 50°C, add sodium bicarbonate aqueous solution (sodium bicarbonate 24g: water 100ml), stir, after bubbles emerge, add dropwise iodine aqueous solution (iodine 23g: water 100ml) containing sodium iodide (20g), continue heating Stir for half an hour. Thin-layer chromatography detects that the reaction is complete. Add sodium persulfate to remove excess iodine. Separate the organic layer with a separatory funnel, dry and concentrate, and recrystallize with 1,2-dichloroethane to obtain 23 g of 3 -Methoxycarbonylmethyl-2-methoxycarbonyl-4-iodo-5-methyl-1H-pyrrole, yield 91%. Mp 102°C. EIMS m/e 336 (M ⁺ ).

Dissolve 3-methoxycarbonylmethyl-2-methoxycarbonyl-4-iodo-5-methyl-1H-pyrrole (23g, 65mmol) in methanol (100ml), heat the oil bath to reflux, stir After adding an aqueous solution of sodium iodide (sodium iodide 11g: water 10ml), concentrated hydrochloric acid (37%) (2ml) was added, iodine was precipitated, and the solution quickly turned dark red. After 15 minutes, sodium persulfate was added to remove Excess iodine, the methanol in the reaction solution was evaporated, extracted with 1,2-dichloroethane (50ml), dried, and the organic layer was concentrated to obtain 12g of 3-methoxycarbonylmethyl-2-methoxy Carbonyl-5-methyl-1H-pyrrole, 87% yield. Mp 238°C. H ¹ NMR (DMSO): 5.98 (s, 1H, 3-pyrrole-H), 3.80 (s, 2H _, CH ₂ CO ₂ Me), 2.3 (s, 3H, CH 3 ), 1.3 (s, 3H, _CO2CH3 ) , 1.2 (s _, 3H , _CO2CH3 ) _.

Phosphorus oxychloride (8ml) was added dropwise to N,N-dimethylformamide (8ml) under an ice bath at 0-5°C, and after stirring for 15 minutes at the same temperature, 12g of 3-methoxy Carbonylmethyl-2-methoxycarbonyl-5-methyl-1H-pyrrole (53mmol) in 1,2-dichloroethane (50ml) was stirred at 0-5°C for 15 minutes, then removed In an ice bath, heat to 50°C with an oil bath. After stirring for half an hour, add an aqueous solution of sodium acetate (5 g of anhydrous sodium acetate: 20 ml of water), stir for half an hour, cool the reaction solution, separate the organic layer with a separatory funnel and concentrate , separated on a silica gel column (R _f =0.5) with an eluent of ethyl acetate:petroleum ether=1:3 to obtain 10 g of 3-methoxycarbonylmethyl-2-methoxycarbonyl-4-methyl Acyl-5-methyl-1H-pyrrole, 80% yield. Mp 132°C. H ¹ NMR (CDCl ₃ ): 10.0 (s, 1H, CHO ), 9.1 (br, 1H, NH), 4.3 (q, 2H, CO ₂ CH ₂ CH ₃ ), 2.3 (s, 3H, C H ₃ ), 1.3(t, 3H, CO ₂ CH ₃ ), 1.2(t, 3H, CO ₂ CH ₃ ).

3-Methoxycarbonylmethyl-2-methoxycarbonyl-4-formyl-5-methyl-1H-pyrrole (0.86g, 3.6mmol) and 5-bromo-(1H)-indole-2 - Ketone (0.82g, 3.6mmol) was mixed and stirred in 20ml ethanol, and the oil bath was heated to 80°C. Add hexahydropyridine (0.1g) dropwise, react for 3 hours and then concentrate the reaction solution, add 1,2-dichloroethane (10ml), wash with 20ml of water and 20ml of 2N hydrochloric acid, concentrate the organic layer, and wash with ethyl acetate:petroleum Ether = 1:2 eluent was purified on a silica gel column ( _Rf = 0.5) to obtain 1.16 g of yellow solid (E)-3-[(3-methoxycarbonylmethyl-2-methoxycarbonyl-5 -Methyl-1H-pyrrol-4-yl)methylene]-(1H)-indol-2-one, yield 91%. Mp215°C.

H ¹ NMR (CDCl ₃ ): 9.10 (s, 1H, NH), 7.70 (s, 1H, vinyl-H), 7.38 (t, 2H, Ph-H), 7.19 (s, 1H, Ph-H), 6.78(t, 2H, Ph-H), 3.90( s , 3H, _{CO2CH2CH3 )} , 3.88(s, _{2H ,} CO2CH2CH3 ₎ , 3.67(s, _3H , _CO2C H3 ) _, 2.20 (s _, 3H, CH3 ). EIMS m/e 354 (M ⁺ ).

Example 4 (E)-5-bromo-3-[(3-methoxycarbonylmethyl-2-methoxycarbonyl-5-methyl-1H-pyrrol-4-yl)methylene]-( The preparation of 1H)-indol-2-one (compound I-4):

3-Methoxycarbonylmethyl-2-methoxycarbonyl-4-formyl-5-methyl-1H-pyrrole (0.95g, 3.6mmol) and 5-bromo-(1H)-indole-2 - Ketone (0.8g, 3.6mmol) was mixed and stirred in 20ml ethanol, heated to 80°C in an oil bath, hexahydropyridine (0.1g) was added dropwise, and the reaction solution was concentrated after 2.5 hours of reaction, and 1,2-dichloroethane was added (10ml), washed with 20ml of water and 20ml of 2N hydrochloric acid, the organic layer was concentrated, and purified on a silica gel column with an eluent of ethyl acetate:petroleum ether=1:4 ( _Rf =0.6) to obtain 1.4g of a yellow solid (E )-5-bromo-3-[(3-methoxycarbonylmethyl-2-methoxycarbonyl-5-methyl-1H-pyrrol-4-yl)methylene]-(1H)-indole -2-one, yield 91%. Mp 243°C.

H ¹ NMR (CDCl ₃ ): 9.10 (s, 1H, NH), 7.70 (s, 1H, vinyl-H), 7.38 (t, 2H, Ph-H), 7.19 (s, 1H, Ph-H), 6.78(t, 2H _, Ph-H), 3.90(s, 3H, _{CO2CH2CH3 )} , 3.88(s, _2H , CO2CH2CH3 ₎ , 3.67(s, _3H , _CO2C H3 ) _, 2.20 (s _, 3H, CH3 ). EMS m/e 432 (M ⁺ ).

Example 5 (E) 3-[(2,5 dimethyl-2-ethoxycarbonyl-1H-pyrrol-3-yl) methylene]-(1H)-indol-2-one (compound I -5) Preparation:

1H-Indol-2-one (6.5g) was dissolved in concentrated sulfuric acid (98%) (25ml) and fuming nitric acid (20ml) was added dropwise while maintaining the mixture at -10°C to -15°C. After addition of nitric acid (20 ml), the reaction mixture was stirred for half an hour at 0°C and poured into ice water, the precipitate was collected by filtration and recrystallized from 50% acetic acid and the crystalline product was filtered and dried under vacuum to give 6 g of 5 -Nitro-2-1H-indol-2-one, yield 70%.

Acetic acid (10ml) and sodium hydroxide (1.2g, 30mmol) were heated to 80°C. After the sodium hydroxide was completely dissolved, the reaction solution was heated to reflux. Ethyl acetoacetate (4.5 g, 34 mmol) was added to the reactor, and an aqueous solution of sodium nitrite (8 g, 0.11 mol, 10 ml of water) was slowly added dropwise. After the dropwise addition, the temperature was slowly lowered to room temperature, and white crystals precipitated after standing for four hours, which were filtered. The precipitate was dissolved in acetic acid (10ml), heated to 80°C, and acetylacetone (3.4g, 34mmol) and Zn powder (2g) were added. After two hours, the reaction solution was poured into ice water, and a white precipitate precipitated out. After filtration, the excess Zn powder was washed away with acetic acid (10 ml), and dried to obtain 3.6 g of white solid 3,5 dimethyl-2-ethoxycarbonyl-1H-pyrrole, with a yield of 65%. Mp126°C. H ¹ NMR (CDCl ₃ ): 8.76 (br, 1H, NH), 5.79 (s, 1H, CH ), 4.29 (q, 2H, CH ₂ CH ₃ ), 2.30 (s, 3H, CH ₃ ) , 2.24 ( _s , _3H , CH3 ) _, 1.350 (t, 3H, CH2CH3 ).

Phosphorus oxychloride (4ml) was added dropwise to N,N-dimethylformamide (4ml) under ice-cooling at 0-5°C. After stirring at the same temperature for 15 minutes, a solution of 3,5-dimethyl-2-ethoxycarbonyl-1H-pyrrole (3 g, 18 mmol) in 1,2-dichloroethane (20 ml) was slowly added dropwise. After continuing to stir at 0-5°C for 15 minutes, remove the ice bath, heat to 50°C with an oil bath, and continue stirring for half an hour, then add an aqueous solution of sodium acetate (1 g of anhydrous sodium acetate: 10 ml of water). After half an hour, the reaction solution was cooled, and the organic layer was separated with a separatory funnel and concentrated, and separated on a silica gel column with an eluent of ethyl acetate:petroleum ether=1:1 (R _f =0.5) to obtain 2.8 g of a white solid The yield of 3,5-dimethyl-4-formyl-2-ethoxycarbonyl-1H-pyrrole was 80%. Mp 182°C. H ¹ NMR (CDCl ₃ ): 10.0 (s, 1H, CHO ), 9.76 (br, 1H, NH), 4.29 (q, 2H, CH ₂ CH ₃ ), 2.30 (s, 3H, CH ₃ ) _, 2.24 (s, _3H , CH3 ), 1.350 ₍ t, 3H, CH2CH3 ).

3,5 Dimethyl-4-formyl-2-ethoxycarbonyl-1H-pyrrole (1 g, 5.1 mmol) and 2-(1H)-indol-2-one (0.6 g, 5.1 mmol) were mixed in Mix and stir in 20ml of ethanol, heat the oil bath to 80°C, add hexahydropyridine (0.1g) dropwise, react for 4 hours, concentrate the reaction solution, add 1,2-dichloroethane (10ml), add 20ml of water and 20ml of 2N After washing with hydrochloric acid, the organic layer was concentrated, and purified on a silica gel column with an eluent of ethyl acetate:petroleum ether=1:1 (R _f =0.35) to obtain 1.48 g of (E)3-[(3,5di Methyl-2-ethoxycarbonyl-1H-pyrrol-4-yl)methylene]-(1H)-indol-2-one, yield 94%. Mp 185°C.

H ¹ NMR (DMCO): 7.55 (s, 1H, vinyl-H), 6.83-7.20 (m, 4H, Ph-H), 4.35 (q, 2H, CO ₂ CH ₂ CH ₃ ), 2.19 (s, 3H, CH3 ₎ , 2.06 (s, 3H, CH3 ) _, 1.36 ₍ t, 3H _, CO2CH2CHH3 ₎ . EIMS m/e 310 (M ⁺ ).

Example 6 (E)-5-bromo-3-[(3,5 dimethyl-2-ethoxycarbonyl-1H-pyrrol-4-yl)methylene]-(1H)-indole-2 - Preparation of ketone (compound 1-6):

3,5 Dimethyl-4-formyl-2-ethoxycarbonyl-1H-pyrrole (1 g, 5.1 mmol) and 5-bromo-2-(1H)-indol-2-one (1 g, 5.1 mmol) were mixed and stirred in 20ml ethanol, the oil bath was heated to 80°C, hexahydropyridine (0.1g) was added dropwise, the reaction solution was concentrated after 3 hours of reaction, 1,2-dichloroethane (10ml) was added, 20ml water and 20 ml of 2N hydrochloric acid was dried and washed, the organic layer was concentrated, and purified on a silica gel column with an eluent of ethyl acetate:petroleum ether=1:2 (R _f =0.4) to obtain 1.8 g of (E)-5-bromo- 3-[(3,5Dimethyl-2-ethoxycarbonyl-1H-pyrrol-4-yl)methylene]-(1H)-indol-2-one, yield 95%. Mp 213°C.

H ¹ NMR (CDCl ₃ ): 9.08 (s, 1H, NH), 7.81 (s, 1H, NH), 7.72 (s, 1H, vinyl-H), 7.31 (d, 2H, Ph-H), 7.17 ( s, 1H, Ph-H), 6.75 (d, 2H, Ph-H), 4.35 (q, 2H, CO ₂ CH ₂ CH ₃ ), 2.19 (s, 3H, CH ₃ ), 2.06 (s, 3H, CH3 ₎ , 1.36 (t, 3H _{, CO2CH2CH3 )} . EIMS m/e 388 (M ⁺ ).

Example 7 (E)-5-nitro-3-[(3,5 dimethyl-2-ethoxycarbonyl-1H-pyrrol-4-yl)methylene]-(1H)-indole- Preparation of 2-keto (compound I-7):

3,5 Dimethyl-4-formyl-2-ethoxycarbonyl-1H-pyrrole (1g, 5.1mmol) and 5-nitro-2-(1H)-indol-2-one (0.9g , 5.1mmol) mixed and stirred in 20ml ethanol, heated to 80°C in an oil bath, added dropwise hexahydropyridine (0.1g), reacted for 3 hours and concentrated the reaction solution, added 1,2-dichloroethane (10ml), 20ml Water and 20ml of 2N hydrochloric acid were dried and washed, the organic layer was concentrated, and purified on a silica gel column with an eluent of ethyl acetate:petroleum ether=2:1 ( _Rf =0.4) to obtain 1.8g of (E)-5- as a yellow solid Nitro-3-[(3,5 dimethyl-2-ethoxycarbonyl-1H-pyrrol-4-yl)methylene]-(1H)-indol-2-one, yield 95%. Mp 286°C.

H ¹ NMR (DMSO-d6): 8.18 (d, 2H, Ph-H), 7.70 (s, 1H, vinyl-H), 7.65 (s, 1H, Ph-H), 7.05 (d, 2H, Ph-H) H), 4.22 (q, 2H, CO ₂ CH ₂ CH ₃ ), 3.98 (q, 2H, CO ₂ CH ₂ CH ₃ ), 3.77 (s, 2H, CH ₂ CO ₂ CH ₂ CH ₃ ), 2.09 (s, 3H _, CH3 ₎ , 1.36 ₍ t, 3H _, CO2CH2CH3 ), 1.20 ( _t , 3H _, CO2CH2CH3 ₎ . EIMS m/e 355 (M ⁺ ).

Example 8 (E) 3-[(3,5 dimethyl-2-methoxycarbonyl-1H-pyrrol-4-yl)methylene]-(1H)-indol-2-one (compound I -8) Preparation:

Acetic acid (10ml) and sodium hydroxide (1.2g, 30mmol) were heated to 80°C. After the sodium hydroxide was completely dissolved, the reaction solution was heated to reflux. Methyl acetoacetate (4.7 g, 34 mmol) was added to the reactor, and an aqueous solution of sodium nitrite (8 g, 0.11 mol, 10 ml of water) was slowly added dropwise. After the dropwise addition, the temperature was slowly lowered to room temperature, and white crystals precipitated after standing for four hours, which were filtered. The precipitate was dissolved in acetic acid (10ml), heated to 80°C, and acetylacetone (3.4g, 34mmol) and Zn powder (2g) were added. After two hours, the reaction solution was poured into ice water, and a white precipitate precipitated out. The precipitate was filtered, the excess Zn powder was washed away with acetic acid (10 ml), and dried to obtain 3.8 g of white solid 3,5 dimethyl-2-ethoxycarbonyl-1H-pyrrole with a yield of 68%. Mp 76°C. H ¹ NMR (CDCl ₃ ): 8.76 (br, 1H, NH), 5.80 (s, 1H, CH ), 3.82 (s, 3H, CH ₃ ), 2.30 (s, 3H, CH ₃ ), 2.24 ₍ s, 3H, CH3 ).

Phosphorus oxychloride (4ml) was added dropwise into N,N-dimethylformamide (4ml) under ice-cooling at 0-5°C. After stirring at the same temperature for 15 minutes, a solution of 3,5-dimethyl-2-methoxycarbonyl-1H-pyrrole (3.4 g, 18 mmol) in 1,2-dichloroethane (20 ml) was slowly added dropwise. After continuing to stir at 0-5°C for 15 minutes, remove the ice bath, heat to 50°C with an oil bath, and continue stirring for half an hour, then add an aqueous solution of sodium acetate (1 g of anhydrous sodium acetate: 10 ml of water). After half an hour, the reaction solution was cooled, and the organic layer was separated with a separatory funnel and concentrated, and separated on a silica gel column with an eluent of ethyl acetate:petroleum ether=1:1 (R _f =0.6) to obtain 3.1 g of a white solid The yield of 3,5-dimethyl-4-formyl-2-methoxycarbonyl-1H-pyrrole was 82%. H ¹ NMR (CDCl ₃ ): 9.76 (s _{, 1H, CHO), 3.82 (s, 3H, CH 3 ), 2.30 (s, 3H, CH 3} ) , _2.24 ( s, 3H, CH ₃ ).

3,5 Dimethyl-4-formyl-2-methoxycarbonyl-1H-pyrrole (1g, 5.5mmol) and 2-(1H)-indol-2-one (0.7g, 5.1mmol) in 20ml of ethanol was mixed and stirred, and the oil bath was heated to 80°C. Hexahydropyridine (0.1g) was added dropwise, and after reacting for 4 hours, the reaction solution was concentrated, dissolved in 1,2-dichloroethane (10ml), washed with 20ml of water and 20ml of 2N hydrochloric acid, and the organic layer was concentrated. Purification on a silica gel column (R _f =0.55) with ethyl acetate:petroleum ether=1:1 eluent gave 1.46 g of (E)3-[(3,5-dimethyl-2-methoxy) as a yellow solid ylcarbonyl-1H-pyrrol-4-yl)methylene]-(1H)-indol-2-one, yield 94%. Mp213°C.

H ¹ NMR (DMCO): 7.55 (s, 1H, vinyl-H), 6.83-7.20 (m, 4H, Ph- H ), 3.75 (s, CO ₂ CH ₃ ), 2.20 (s, 3H, CH ₃ ), 2.06 (s _, 3H, CH3 ). EIMS m/e 296 (M ⁺ ).

Example 9 (E)-5-nitro-3-[(3,5 dimethyl-2-methoxycarbonyl-1H-pyrrol-4-yl)methylene]-(1H)-indole- Preparation of 2-keto (compound I-9):

3,5 Dimethyl-4-formyl-2-methoxycarbonyl-1H-pyrrole (1g, 5.5mmol) and 5-nitro-2-(1H)-indol-2-one (0.98g , 5.5mmol) were mixed and stirred in 20ml ethanol, and the oil bath was heated to 80°C. Add hexahydropyridine (0.1g) dropwise, react for 3 hours and then concentrate the reaction solution, add 1,2-dichloroethane (10ml), wash with 20ml of water and 20ml of 2N hydrochloric acid, concentrate the organic layer, and wash with ethyl acetate:petroleum Ether = 2:1 eluent was purified on a silica gel column (R _f =0.3) to obtain 1.9 g of (E)-5-nitro-3-[(3,5-dimethyl-2-methoxy) as a yellow solid ylcarbonyl-1H-pyrrol-4-yl)methylene]-(1H)-indol-2-one, yield 95%. Mp 218°C.

H ¹ NMR (DMSO): 8.15 (d, 2H, Ph-H), 7.71 (s, 1H, vinyl-H), 7.66 (s, 1H, Ph-H), 7.00 (d, 2H, Ph-H) , 3.75 (s, 3H, CO ₂ CH ₃ ), 2.20 (s, 3H, CH ₃ ), 2.06 (s, 3H, CH ₃ ). EIMS m/e 341 (M ⁺ ).

Example 10 (E)-5-nitro-3-[(3-ethoxycarbonylmethyl-2-ethoxycarbonyl-5-methyl-1H-pyrrol-4-yl)methylene]- Preparation of (1H)-indol-2-one (compound 1-10):

3-Ethoxycarbonylmethyl-2-ethoxycarbonyl-4-formyl-5-methyl-1H-pyrrole (1g, 3.7mmol) and 5-nitro-2-(1H)-indole -2-Kone (0.78g, 3.7mmol) was mixed and stirred in 20ml of ethanol, and the oil bath was heated to 80°C. Hexahydropyridine (0.1g) was added dropwise, and after 5 hours of reaction, the concentrated reaction solution was added with 1,2-dichloroethane (10ml), 20ml of water and 20ml of 2N hydrochloric acid for drying and washing, and the organic layer was concentrated and washed with ethyl acetate:petroleum ether = 1:1 eluent was purified on a silica gel column (R _f =0.5) to give 1.43 g of (E)-5-nitro-3-[(3-ethoxycarbonylmethyl-2-ethane) as a yellow solid Oxycarbonyl-5-methyl-1H-pyrrol-4-yl)methylene]-(1H)-indol-2-one, yield 91%. Mp 341°C.

H ¹ NMR (DMSO-d6): 8.18 (d, 2H, Ph-H), 7.70 (s, 1H, vinyl-H), 7.65 (s, 1H, Ph-H), 7.05 (d, 2H, Ph-H) H), 4.22 (q, 2H, CO ₂ CH ₂ CH ₃ ), 3.98 (q, 2H, CO ₂ CH ₂ CH ₃ ), 3.77 (s, CH 2 CO _{2 CH 2 CH 3} ), 2.09 ( s, 3H _, CH3 ₎ , 1.36 (t, _3H , CO2CH2CH3 ₎ , 1.20 (t, 3H _, CO2CH2CH3 _{) .} EIMS m/e 427 (M ⁺ ).

Example 11 (E)-5-carboxy-3-[(3-ethoxycarbonylmethyl-2-ethoxycarbonyl-5-methyl-1H-pyrrol-4-yl)methylene]-( The preparation of 1H)-indol-2-one (compound I-11):

(1H)-Indol-2-one (6.7g, 50mmol) was added to a suspension of 23g of aluminum trichloride in 30ml of 1,2-dichloroethane, and chloroacetyl chloride (11.3g) was added slowly And emit hydrogen chloride gas. After stirring for 10 minutes, the reaction solution was heated to 40 to 50°C. Thin layer chromatography (ethyl acetate, silica gel) showed no remaining starting material. The mixture was cooled to room temperature and poured into ice water The precipitate was collected by vacuum filtration, washed with water (80ml) and dried under vacuum to give 10.3g of 5-chloroacetyl-(1H)-indol-2-one as a white solid , yield 98%.

A suspension of 5-chloroacetyl-(lH)-indol-2-one (9.3 g, 49 mmol) in pyridine (9 g) was stirred at 80 to 90°C for 3 hours and then cooled to room temperature. The precipitate was collected by vacuum filtration and washed with ethanol (20ml). The solid was dissolved in 90 ml of 2.5N sodium hydroxide and stirred at 70 to 80°C for 3 hours. The mixture was cooled to room temperature and acidified to pH=2 with 0.5N hydrochloric acid. The precipitate was collected by vacuum filtration and washed thoroughly with water (100ml) to give 5-carboxy-2-(lH)-indol-2-one as a dark brown solid. After standing the filtrate overnight, 2 g of 5-carboxy-(1H)-indol-2-one were obtained as a yellow solid. The dark brown product was dissolved in hot ethanol (20ml), the insolubles were removed by filtration and the filtrate was concentrated to give 5-carboxy-2-(1H)-indol-2-one as 5.6g of a brown solid. The combined yield was 97%. Mp 286°C. H ¹ NMR (DMSO): 12.56 (br, 1H, COO H ), 10.70 (s, 1H, NH-1), 7.81 (dd, 1H, Ph-H), 7.74 (s, 1H, Ph-H), 3.53 (s _, 2H, CH2 ).

3-Ethoxycarbonylmethyl-2-ethoxycarbonyl-4-formyl-5-methyl-1H-pyrrole (1g, 3.7mmol) and 5-carboxy-2-(1H)-indole- 2-Kone (0.65g, 3.7mmol) was mixed and stirred in 20ml of ethanol, and the oil bath was heated to 80°C. Add hexahydropyridine (0.1g) dropwise, react for 5 hours, concentrate the reaction solution, dissolve it with 1,2-dichloroethane (10ml), wash with 20ml of water and 20ml of 2N hydrochloric acid, concentrate the organic layer, wash with ethyl acetate : Petroleum ether = 4: 1 eluent was purified on a silica gel column (R _f =0.25) to obtain 1.49 g of (E)-5-carboxy-3-[(3-ethoxycarbonylmethyl-2) as a red solid -Ethoxycarbonyl-5-methyl-1H-pyrrol-4-yl)methylene]-(1H)-indol-2-one, yield 95%. Mp 214°C.

H ¹ NMR (DMSO-d6): 7.82 (d, 2H, Ph-H), 7.56 (s, 1H, vinyl-H), 7.54 (s, 1H, Ph-H), 6.98 (q, 2H, Ph-H) H), 4.21 (q, 2H, CO ₂ CH ₂ CH ₃ ), 3.98 (q, 2H, CO ₂ CH ₂ CH ₃ ), 3.77 (s, 2H, CH ₂ CO ₂ CH ₂ CH ₃ ), 2.09 (s, 3H _, CH3 _{) ,} 1.36 (t _, 3H, CO2CH2CH3 ), _1.20 (t, 3H _, CO2CH2CH3 ₎ . EIMS m/e 426 (M ⁺ ).

Example 12 (E)-5-aminosulfonyl-3-[(3-ethoxycarbonylmethyl-2-ethoxycarbonyl-5-methyl-1H-pyrrol-4-yl)methylene] Preparation of -(1H)-indol-2-one (compound I-12):

To a 100ml flask containing 27ml of chlorosulfonic acid was slowly added 13.3g of (1H)-indol-2-one. The reaction temperature was maintained below 30°C during the addition. After the addition, the reaction mixture was stirred at room temperature for 1.5 hours, heated to 68 °C, reacted for 1 hour, cooled, and poured into water, the precipitate was washed with water (50 ml) and dried in a vacuum oven to obtain 11 g of 5-chlorosulfur Acyl-(1H)-indol-2-one, 50% yield.

5-Chlorosulfonyl-(1H)-indol-2-one (2.1 g) was added to aqueous ammonia (10 ml) in 10 mL of ethanol and stirred overnight at room temperature. The mixture was concentrated and the solid was collected by vacuum filtration to afford 0.4 g of 5-aminosulfonyl-(lH)-indol-2-one as an off-white solid, 20% yield. Mp 125°C. H ¹ NMR (DMSO): 10.67 (s, 1H, NH -1), 7.63-7.66 (m, 2H, Ph-H), 7.13 (s, 2H, H ₂ N SO ₂ -5), 6.91 (d, 1H, Ph-H), 3.56 ( _s , 2H, CH2 ). MS m/e 341 (M ⁺ ).

3-Ethoxycarbonylmethyl-2-ethoxycarbonyl-4-formyl-5-methyl-1H-pyrrole (1g, 3.7mmol) and 5-aminosulfonyl-(1H)-indole- 2-Kone (0.543g, 3.7mmol), mixed and stirred in 20ml ethanol, heated to 80°C in an oil bath, added hexahydropyridine (0.1g) dropwise, reacted for 5 hours, concentrated the reaction solution, and used ethyl acetate:petroleum ether =1:3 eluent was purified on a silica gel column (R _f =0.85) to obtain 1.2 g of (E)-5-aminosulfonyl-3-[(3-ethoxycarbonylmethyl-2) as an orange-yellow solid -Ethoxycarbonyl-5-methyl-1H-pyrrol-4-yl)methylene]-(1H)-indol-2-one, yield 70%. Mp 148-150°C.

H ¹ NMR (CDCl ₃ ): 7.60(s, 1H, vinyl-H), 7.59(d, 1H, Ph-H), 7.22(s, 1H, Ph-H), 7.06(d, 1H, Ph-H ), 4.25(q, 2H, CO ₂ CH ₂ CH ₃ ), 3.98(q, 2H, CO ₂ CH ₂ CH ₃ ), 3.80(s, 2H, CH 2 CO _{2 CH 2 CH 3} ), 2.10( s, 3H, CH3 ₎ , 1.28 (t _{, 3H ,} CO2CH2CH3 ), 1.05 (t, 3H _, CO2CH2CH3 _{) .} EIMS m/e 461 (M ⁺ ).

Example 13 (E)-5-methylaminosulfonyl-3-[(3-ethoxycarbonylmethyl-2-ethoxycarbonyl-5-methyl-1H-pyrrol-4-yl)methylene Base]-(1H)-indol-2-one (compound I-13) preparation:

A suspension of 5-chlorosulfonyl-2-(1H)-indol-2-one (2.3g, 10mmol) in methylamine (10ml) in methanol (10ml) was stirred at room temperature for 4 hours during which time white solid. The precipitate was collected by filtration, washed with sodium hydroxide (5ml, 1N) and hydrochloric acid (5ml, 1N) and dried in vacuo overnight to give 2.2g of an orange solid 5-methylaminosulfonyl-2-(1H)-indole-2- Ketone, yield 82%. EIMS m/e 225 (M ⁺ -1).

3-Ethoxycarbonylmethyl-2-ethoxycarbonyl-4-formyl-5-methyl-1H-pyrrole (1g, 3.7mmol) and 5-methylaminosulfonyl-2-(1H) -Indol-2-one (0.9g, 3.7mmol) was mixed and stirred in 20ml of ethanol, and the oil bath was heated to 80°C. Hexahydropyridine (0.01g) was added dropwise, and after reacting for 6 hours, the reaction solution was concentrated and purified on a silica gel column with an eluent of ethyl acetate:petroleum ether=1:4 ( _Rf =0.55) to obtain 1.08g of a yellow solid (E)-5-methylaminosulfonyl-3-[(3-ethoxycarbonylmethyl-2-ethoxycarbonyl-5-methyl-1H-pyrrol-4-yl)methylene]- (1H)-Indol-2-one, yield 67%. Mp126°C.

H ¹ NMR (CDCl ₃ ): 9.31 (s, 1H, NH), 8.41 (s, 1H, NH), 7.8 (s, 1H, vinyl-H), 7.63 (d, 1H, Ph-H), 7.42 ( s, 1H, Ph-H), 7.00 (d, 1H, Ph-H), 4.32 (q, 2H, CO ₂ CH ₂ CH ₃ ), 4.10 (q, 2H, CO ₂ CH ₂ CH ₃ ), 3.80(s, 2H _{, CH2CO2CH2CH3} ), 2.42 ₍ s, 3H, NCH3 ) , 2.10 ₍ s, 3H _, CH3 ) , 1.36(t, _{3H , CO2CH2CH 3} ) _, 1.20 ( t , 2H, _CO2CH2CH3 ) _. EIMS m/e 475 (M ⁺ ).

Example 14 (E)-5-(N-morpholinyl)aminosulfonyl-3-[(3-ethoxycarbonylmethyl-2-ethoxycarbonyl-5-methyl-1H-pyrrole-4 Preparation of -(yl)methylene]-(1H)-indol-2-one (compound I-14):

Dissolve 5-chlorosulfonyl-2-(1H)-indol-2-one (2.3g, 10mmol) in tetrahydrofuran (10ml) at room temperature, add morpholine (750mg, 10mmol), stir for 4 hours, A white solid formed during this time. The precipitate was collected by filtration, washed with sodium hydroxide (5ml, 1N) and hydrochloric acid (5ml, IN) and dried under vacuum overnight to give 2.53g of 5-(N-morpholino)sulfamoyl-(1H)-indole as a white solid Indol-2-one, the yield is 90%. EIMS m/e 282 (M ⁺ ).

3-Ethoxycarbonylmethyl-2-ethoxycarbonyl-4-formyl-5-methyl-1H-pyrrole (1g, 3.7mmol) and 5-(N-morpholinyl)aminosulfonyl- (1H)-Indol-2-one (1.03g, 3.7mmol) was mixed and stirred in 20ml of ethanol, and the oil bath was heated to 80°C. Hexahydropyridine (0.01g) was added dropwise, and after reacting for 6 hours, the reaction solution was concentrated and purified on a silica gel column with an eluent of ethyl acetate:petroleum ether=1:4 ( _Rf =0.75) to obtain 1.21g of a yellow solid (E)-5-(N-morpholinyl)aminosulfonyl-3-[(3-ethoxycarbonylmethyl-2-ethoxycarbonyl-5-methyl-1H-pyrrol-4-yl) Methylene]-(1H)-indol-2-one, 62% yield. Mp185°C.

H ¹ NMR (CDCl ₃ ): 9.31 (s, 1H, NH), 8.41 (s, 1H, NH), 7.80 (s, 1H, vinyl-H), 7.63 (d, 1H, Ph-H), 7.42 ( s, 1H, Ph-H), 7.00 (d, 1H, Ph-H), 4.32 (q, 2H, CO ₂ CH ₂ CH ₃ ), 4.10 (q, 2H, CO ₂ CH ₂ CH ₃ ), 3.80 (s, 2H, CH ₂ CO ₂ CH ₂ CH ₃ ), 3.67 (t, 2H, 1.4-morpholinyl), 2.90 (t, 2H, 2.3-morpholinyl), 2.10 (s, 3H, CH ₃ ), 1.36 (t, 3H _, CO2CH2CH3 ), 1.20 ₍ t _, 2H _{, CO2CH2CH3 )} . EIMS m/e 531 (M ⁺ ).

Example 15 (E)-5-(3-chlorophenyl)aminosulfonyl-3-[(3-ethoxycarbonylmethyl-2-ethoxycarbonyl-5-methyl-1H-pyrrole-4 Preparation of -(yl)methylene]-(1H)-indol-2-one (compound 1-15):

5-Chlorosulfonyl-(1H)-indol-2-one (2.1 g, 9 mmol), 3-chloroaniline (1.8 g, 14 mmol) and pyridine (1.4 g) were dissolved in 20 mL of dichloromethane at room temperature The suspension was stirred for 4 hours. The formed precipitate was collected by vacuum filtration, washed twice with 15 ml of water and dried overnight at 40°C under vacuum to give 2.5 g of 5-(3-chlorophenyl)aminosulfonyl-(1H)-indole- 2-Kone, yield 86%. EIMS m/e 321 (M ⁺ -1).

3-Ethoxycarbonylmethyl-2-ethoxycarbonyl-4-formyl-5-methyl-1H-pyrrole (1g, 3.7mmol) and 5-(3-chlorophenyl)aminosulfonyl- (1H)-Indol-2-one (1.19g, 3.7mmol) was mixed and stirred in 20ml of ethanol, and the oil bath was heated to 80°C. Hexahydropyridine (0.01g) was added dropwise, and after reacting for 6 hours, the reaction solution was concentrated and purified on a silica gel column with an eluent of ethyl acetate:petroleum ether=1:5 ( _Rf =0.8) to obtain 1.52g of a yellow solid (E)-5-(3-chlorophenyl)aminosulfonyl-3-[(3-ethoxycarbonylmethyl-2-ethoxycarbonyl-5-methyl-1H-pyrrol-4-yl) Methylene]-(1H)-indol-2-one, 72% yield. Mp 118-120°C.

H ¹ NMR (DMSO): 7.56 (m, 2H, Ph-H), 7.38 (s, 1H, vinyl-H), 6.80-6.98 (m, 5H, Ph-H), 4.21 (q, 2H, CO ₂ CH ₂ CH ₃ ), 3.98 (q, 2H, CO ₂ CH ₂ CH ₃ ), 3.77 (s _{, 2H, CH 2 CO 2 CH 2} CH _{3 ) ,} 2.00 (s, 3H, CH ₃ ), _1.36 (t , 3H _, CO2CH2CH3 ), 1.20(t _, 3H _{, CO2CH2CH3 )} . EIMS m/e 571 (M ⁺ ).

Example 16 (E)-5-(3-fluorophenyl)aminosulfonyl-3-[(3-ethoxycarbonylmethyl-2-ethoxycarbonyl-5-methyl-1H-pyrrole-4 Preparation of -(yl)methylene]-(1H)-indol-2-one (compound I-16):

3-Ethoxycarbonylmethyl-2-ethoxycarbonyl-4-formyl-5-methyl-1H-pyrrole (1g, 3.7mmol) and 5-(3-fluorophenyl)aminosulfonyl- (1H)-Indol-2-one (1.13g, 3.7mmol) was mixed and stirred in 20ml of ethanol, and the oil bath was heated to 80°C. Hexahydropyridine (0.01g) was added dropwise, and after reacting for 6 hours, the reaction solution was concentrated and purified on a silica gel column with an eluent of ethyl acetate:petroleum ether=1:4 ( _Rf =0.85) to obtain 1.49g of a yellow solid (E)-5-(3-fluorophenyl)aminosulfonyl-3-[(3-ethoxycarbonylmethyl-2-ethoxycarbonyl-5-methyl-1H-pyrrol-4-yl) Methylene]-(1H)-indol-2-one, yield 73%. Decompose at Mp>130°C.

H ¹ NMR (DMSO): 7.55 (m, 2H, Ph-H), 7.39 (s, 1H, vinyl-H), 6.80-6.98 (m, 5H, Ph-H), 4.21 (q, 2H, CO ₂ CH ₂ CH ₃ ), 3.98 (q, 2H, CO ₂ CH ₂ CH ₃ ), 3.77 (s _{, 2H, CH 2 CO 2 CH 2} CH _{3 ) ,} 2.00 (s, 3H, CH ₃ ), 1.36 ₍ t _, CO2CH2CH3 ) _, 1.20( _t , 3H _, CO2CH2CH3 ₎ . EIMS m/e 555 (M ⁺ ).

Example 17 (E)-5-dimethylaminosulfonyl-3-[(3-ethoxycarbonylmethyl-2-ethoxycarbonyl-5-methyl-1H-pyrrol-4-yl)ylidene Preparation of methyl]-(1H)-indol-2-one (compound I-17):

2.3 g of 5-chlorosulfonyl-2-(1H)-indol-2-one were added to a suspension of dimethylamine in methanol (10 mL, 2M) at room temperature and stirred for 4 hours, during which time a white solid formed . The precipitate was collected by vacuum filtration, washed with sodium hydroxide (5ml, 1N) and hydrochloric acid (5ml, IN) and dried under vacuum overnight to give 1.8g of 5-dimethylamino-sulfonyl-2-(1H)-indole -2-one, yield 77%. EIMS m/e 240 (M ⁺ ).

3-Ethoxycarbonylmethyl-2-ethoxycarbonyl-4-formyl-5-methyl-1H-pyrrole (1 g, 3.7 mmol) and 5-dimethylamino-sulfonyl-2-( 1H)-Indol-2-one (0.94g, 3.7mmol) was mixed and stirred in 20ml of ethanol, and the oil bath was heated to 80°C. Hexahydropyridine (0.01g) was added dropwise, and after reacting for 6 hours, the reaction solution was concentrated and purified on a silica gel column with an eluent of ethyl acetate:petroleum ether=1:4 ( _Rf =0.60) to obtain 1.17g of a yellow solid (E)-5-Dimethylaminosulfonyl-3-[(3-ethoxycarbonylmethyl-2-ethoxycarbonyl-5-methyl-1H-pyrrol-4-yl)methylene] -(1H)-indol-2-one, yield 65%. Mp145°C.

H ¹ NMR (CDCl ₃ ): 9.32 (s, 1H, NH), 8.43 (s, 1H, NH), 7.80 (s, 1H, vinyl-H), 7.63 (d, 1H, Ph-H), 7.42 ( s, 1H, Ph-H), 7.00 (d, 1H, Ph-H), 4.32 (q, 2H, CO ₂ CH ₂ CH ₃ ), 4.10 (q, 2H, CO ₂ CH ₂ CH ₃ ), 3.80 (s, 2H, CH 2 CO ₂ CH ₂ CH ₃ ), 2.43 (s, _6H , N( CH ₃ ) ₂ ), 2.10 (s, 3H, CH ₃ ), 1.36 (t, 3H, CO _{2 CH2CH3 )} , _1.20 (t _, 2H, CO2CH2CH3 ₎ . EIMS m/e 489 (M ⁺ ).

Example 18 (E)-5-(3-fluoro-4-chlorophenyl)aminosulfonyl-3-[(3-ethoxycarbonylmethyl-2-ethoxycarbonyl-5-methyl-1H Preparation of -pyrrol-4-yl)methylene]-(1H)-indol-2-one (compound 1-18):

At room temperature, 5-chlorosulfonyl-(1H)-indol-2-one (2.1g, 9mmol), 3-fluoro-4-chloroaniline (2.03g, 14mmol) and pyridine (1.4g) were dissolved in 20ml Stir in dichloromethane for 4 hours. Filtration, the precipitate was washed twice with 15 ml of water and dried under vacuum at 40° C. overnight to obtain 2.5 g of pink solid 5-(3-fluoro-4-chlorophenyl)aminosulfonyl-(1H)-indole-2- Ketone, 80% yield. EIMS m/e 340 (M ⁺ ).

3-Ethoxycarbonylmethyl-2-ethoxycarbonyl-4-formyl-5-methyl-1H-pyrrole (1g, 3.7mmol) and 5-(3-fluoro-4-chlorophenyl) Aminosulfonyl-(1H)-indol-2-one (1.25g, 3.7mmol) was mixed and stirred in 20ml of ethanol, and the oil bath was heated to 80°C. Hexahydropyridine (0.01g) was added dropwise, and after reacting for 6 hours, the reaction solution was concentrated and purified on a silica gel column with an eluent of ethyl acetate:petroleum ether=1:4 ( _Rf =0.75) to obtain 1.52g of a yellow solid ( E)-5-(3-fluoro-4-chlorophenyl)aminosulfonyl-3-[(3-ethoxycarbonylmethyl-2-ethoxycarbonyl-5-methyl-1H-pyrrole-4 -yl)methylene]-(1H)-indol-2-one, yield 70%. Decompose at Mp>130°C.

H ¹ NMR (DMSO): 7.56 (m, 2H, Ph-H), 7.38 (s, 1H, vinyl-H), 6.87-6.98 (m, 4H, Ph-H), 4.21 (q, 2H, CO ₂ CH ₂ CH ₃ ), 3.98 (q, 2H, CO ₂ CH ₂ CH ₃ ), 3.77 (s _{, 2H, CH 2 CO 2 CH 2} CH _{3 ) ,} 2.00 (s, 3H, CH ₃ ), _{1.36(t, 3H, CO2CH2CH3 ) ,} 1.20 ( _t , CO2CH2CH3 _{) .} EIMS m/e 589 (M ⁺ ).

Example 19 (E)-5-(hexahydropyridyl)aminosulfonyl-3-[(3-ethoxycarbonylmethyl-2-ethoxycarbonyl-5-methyl-1H-pyrrole-4- Base) methylene]-(1H)-indol-2-one (compound I-19) preparation:

Dissolve 5-chlorosulfonyl-2-(1H)-indol-2-one (2.3g, 10mmol) in tetrahydrofuran (10ml) at room temperature, add hexahydropyridine (750mg, 10mmol), and stir for 4 hours , during which a white solid was formed. The precipitate was collected by filtration, washed with sodium hydroxide (5ml, 1N) and hydrochloric acid (5ml, 1N) and dried under vacuum overnight to give 2.53g of white solid 5-(N-hexahydropyridyl)sulfamoyl-(1H)-ind Indol-2-one, the yield is 90%. EIMS m/e 282 (M ⁺ ).

3-Ethoxycarbonylmethyl-2-ethoxycarbonyl-4-formyl-5-methyl-1H-pyrrole (1g, 3.7mmol) and 5-(N-hexahydropyridyl)aminosulfonyl -(1H)-Indol-2-one (1.03g, 3.7mmol) was mixed and stirred in 20ml of ethanol, and the oil bath was heated to 80°C. Hexahydropyridine (0.01g) was added dropwise, and after reacting for 6 hours, the reaction solution was concentrated and purified on a silica gel column with an eluent of ethyl acetate:petroleum ether=1:1 ( _Rf =0.70) to obtain 1.21g of a yellow solid ( E)-5-(hexahydropyridyl)aminosulfonyl-3-[(3-ethoxycarbonylmethyl-2-ethoxycarbonyl-5-methyl-1H-pyrrol-4-yl)methylene Base]-(1H)-indol-2-one, yield 60%. Mp145°C.

H ¹ NMR (DMCO): 11.20 (s, 1H, NH), 9.99 (s, 1H, NH), 7.70 (s, 1H, vinyl-H), 7.62 (d, 2H, Ph-H), 6.42 (s , 1H, Ph-H), 7.17 (q, 2H, Ph-H), 4.28 (q, 2H, CO ₂ CH ₂ CH ₃ ), 4.00 (q, 2H, CO ₂ CH ₂ CH ₃ ), 3.95 (s, 2H, CH ₂ CO ₂ CH ₂ CH ₃ ), 2.11 (s, 3H, CH ₃ ), 1.60 (m, 6H, 4.5-(CH 2 _{) 3} 3-hexapyridinyl), 1.42 (m, 4H, 1.2-( CH2 _{) 2} -hexapyridinyl), 1.30 ( _t , 3H _, CO2CH2CH3 _{) ,} 1.15(t, 3H _, CO2CH2CH3 ₎ . EIMS m/e 530 (M ⁺ +1).

Example 20 (E)-5-(3-bromophenyl)aminosulfonyl-3-[(3-ethoxycarbonylmethyl-2-ethoxycarbonyl-5-methyl-1H-pyrrole-4 Preparation of -(yl)methylene]-(1H)-indol-2-one (compound 1-20):

At room temperature, 5-chlorosulfonyl-(1H)-indol-2-one (2.1g, 9mmol), 3-bromoaniline (2.3g, 14mmol) and pyridine (1.4g) were dissolved in 20ml of dichloromethane Stir for 4 hours. After filtration, the precipitate was washed twice with 15 ml of water and dried under vacuum at 40°C overnight to obtain 2.9 g of 5-(3-bromophenyl)sulfamoyl-(1H)-indol-2-one as a pink solid, producing Yield 82%, EIMS m/e 365 (M ⁺ ).

3-Ethoxycarbonylmethyl-2-ethoxycarbonyl-4-formyl-5-methyl-1H-pyrrole (1g, 3.7mmol) and 5-(3-bromophenyl)aminosulfonyl- (1H)-Indol-2-one (1.39g, 3.7mmol) was mixed and stirred in 20ml of ethanol, and the oil bath was heated to 80°C. Hexahydropyridine (0.01g) was added dropwise, and after reacting for 6 hours, the reaction solution was concentrated and purified on a silica gel column with an eluent of ethyl acetate:petroleum ether=1:5 ( _Rf =0.65) to obtain 1.72g of a yellow solid ( E)-5-(3-bromophenyl)aminosulfonyl-3-[(3-ethoxycarbonylmethyl-2-ethoxycarbonyl-5-methyl-1H-pyrrol-4-yl)ylidene Methyl]-(1H)-indol-2-one, yield 73%. Mp 134-138°C.

H ¹ NMR (DMSO): 7.56 (m, 2H, Ph-H), 7.38 (s, 1H, vinyl-H), 6.80-6.98 (m, 5H, Ph-H), 4.21 (q, 2H, CO ₂ CH ₂ CH ₃ ), 3.98 (q, 2H, CO ₂ CH ₂ CH ₃ ), 3.77 (s _{, 2H, CH 2 CO 2 CH 2} CH _{3 ) ,} 2.00 (s, 3H, CH ₃ ), _1.36 (t , 3H _, CO2CH2CH3 ), 1.20(t _, 3H _{, CO2CH2CH3 )} . EIMS m/e 615 (M ⁺ ).

Example 21 Experiment of the inhibitory effect of the compounds of the present invention on cyst lining epithelial cells of human autosomal dominant polycystic kidney disease (ADPKD):

1. Reagents and instruments

Human polycystic kidney disease cyst lining epithelial cell line: established and preserved by the Nephrology Laboratory of Shanghai Changzheng Hospital;

DMEM: F12 (1:1) culture medium (GIBCO company);

Calf serum (Changzhou Sijiqing Company);

Recombinant human epidermal growth factor (rhEGF) (Calbiochem Company);

Microplate reader (LabsysteEIMS MK3)

Compounds to be tested: compounds I-1, I-2, I-3, I-4, I-5, I-6, I-7, I-8, I-9, I prepared according to Example 1-16 -10, I-11, I-12, I-13, I-14, I-15, I-16.

The compound to be tested is diluted with culture medium, and the concentration gradient is 50μM, 5μM, 500nM, 50nM, 5nM, 0.5nM, 0.05nM

2. Experimental method

Cell proliferation was measured by MTT assay. Cyst lining epithelial cells in the logarithmic growth phase were seeded in 96-well plates (1×103/well). After 24 hours, the serum-free medium was changed for 24 hours to make the cells rest in the resting phase. Discard the supernatant and replace with drug solutions of various concentrations containing 2% calf serum. Set up 9 duplicate wells for each dose group. After incubation at 37°C for 72 hours, each dose group was stimulated with 1ng/ml rhEGF for 24 hours (determined according to the preliminary experiment). optimal stimulating concentration). A blank control group with no drug treatment and no EGF stimulation was set up, and only culture solution (2% calf serum) was added all the time. Take 6 duplicate wells, add 10 μl MTT to each well, discard the supernatant after incubation for 4 hours, add 100 μl DMSO to each well to dissolve the crystals, read the absorbance value at 490 nm with a microplate reader, and repeat the experiment 5 times.

Inhibition rate=(A490 control well-A490 administration well)/A490 control well×100%

According to the inhibition rate of each concentration, the logarithm of the concentration of the competitor was linearly regressed with Logit [I] to obtain the IC ₅₀ of the concentration of the competitor when 50% of the conjugate was inhibited. Each of the above experiments was repeated twice, and the average IC ₅₀ value of the two experiments was obtained as the final index of the inhibitory ability, and the inhibition percentage of the test compound was measured.

3. Experimental results

The experimental results are shown in Table 1.

Table 1. Compound of the present invention is to the inhibitory percentage (%) of human polycystic kidney disease cyst lining epithelial cell at each concentration

Example 22 Experiment of the inhibitory effect of the compounds of the present invention on A-549 human lung adenocarcinoma cells, A-431 human skin squamous cell carcinoma cells and MDA-MB-468 human breast cancer cells

1. Reagents and instruments

RPMII640 was purchased from Gibco; sulforhodamine B was purchased from Sigma; trichloroacetic acid (TCA), acetic acid (HAC) and Tris base unbuffer were all of domestic analytical grade; microplate reader (VERSAmax).

Compound to be tested: compounds I-1, I-2, I-3, I-4, I-5, I-6, I-7, I-8, I-9, I prepared according to Example 1-20 -10, I-11, I-12, I-13, I-14, I-15, I-16, I-17, I-18, I-19, I-20.

The compound to be tested is diluted with DMSO, the concentration gradient is 10 ^-4 M, 10 ^-5 M, 10 ^-6 M

2. Experimental method (sulfonylrhodamine B (SRB) protein staining method)

According to the cell growth rate, A-549 human lung adenocarcinoma cells, A-431 human skin squamous cell carcinoma cells and MDA-MB-468 human breast cancer cells in the logarithmic growth phase were seeded in 96-well culture plates at 90 μl/well , grow adherently for 24 hours, and then add 10 μl/well of drug. Three replicate wells were set up for each concentration. And set the corresponding concentration of normal saline vehicle control and no cell apoptosis well. Tumor cells were cultured at 37°C and 5% CO ₂ for 72 hours, then the culture medium was discarded, the cells were fixed with 10% cold TCA, and after 1 hour at 4°C, they were washed 5 times with distilled water and dried naturally in the air. Then add 100 μl/well of SRB (Sigma) 4 mg/ml solution prepared by 1% glacial acetic acid, stain at room temperature for 15 minutes, remove the supernatant, wash 5 times with 1% acetic acid, air-dry, and finally add 150 μl/well Tris solution, the A value was measured at a wavelength of 540nm with a microplate reader.

The inhibition rate of tumor cell growth was calculated by the following formula:

Inhibition rate=(A540 control well-A540 administration well)/A540 control well×100%

3. Experimental results

The experimental results are shown in Table 2-4.

Table 2. The inhibitory rate (%) of compound of the present invention to A-549 human lung adenocarcinoma cell growth

Table 3. The inhibitory rate (%) of compound of the present invention to the growth of A-431 human skin squamous cell carcinoma cells

Table 4. Compound of the present invention is to the inhibitory rate (%) of MDA-MB-468 human breast cancer cell growth

It can be seen from Tables 1-4 that the compounds of the present invention have higher inhibitory activity on EGFR-rich tumor cells. IC of eight compounds (compounds I-4, I-5, I-15, I-6, I-12, I-3, I-9 and I-11) in the growth inhibition assay of ADPKD cyst lining epithelial cells ₅₀ reached 10 ^-5 M, the IC ₅₀ of two compounds reached 10 ^-6 M, and the IC ₅₀ of compound I-4 reached 10 ^-7 M. The effects of two compounds (I-7 and I-8) on A-549 human lung adenocarcinoma cells, A-431 human skin squamous cell carcinoma cells and MDA-MB-468 human breast cancer cells at a concentration of 10 ^-6 M The percent inhibition was 86%, showing a potent inhibitor.

Claims (10)

1. Substituted 1H-indol-2-one compounds having the following general formula (I):

in: _R is selected from hydrogen, C _1-5 alkyl, C _2-5 alkenyl, C _3-10 cycloalkyl, hydroxyl, C _1-5 alkoxy, acetyl, carboxyl, amido, thioamide group, sulfonyl group and trihalomethylsulfonyl group; R ₂ , R ₃ , R ₄ , and R ₅ are each independently selected from hydrogen, C _1-5 alkyl, C _1-4 alkoxy, aryl, aryloxy, C _1-5 alkylaryl, C _1-5 alkyl aryloxy, C _3-10 cycloalkyl, mercapto, C _1-5 alkyl mercapto, aryl mercapto, sulfinyl, halogen, trihalomethyl, amino, cyano, nitro, carboxyl, Oxycarboxyl, amido, thioamido, sulfonamide, trihalomethylsulfonyl, N-sulfinamide, S-sulfinamide, trihalomethylsulfinyl, O-carbamoyl, O-thiocarbamoyl and -NR ₁₁ R ₁₂ , wherein R ₃ and R ₄ , R ₄ and R ₅ can combine to form a six-membered aromatic ring, methylenedioxy or ethylenedioxy; R ₄ and R ₅ can also be selected from the following groups simultaneously or separately:

Wherein _R is selected from hydrogen; C _1-5 alkyl; C _1-5 alkoxy; aryl; aryloxy; C _1-5 alkylaryl; C _1-5 alkylaryloxy; trihalogen Methyl; pyridyl; pyrimidinyl; thienyl; each of the above groups may be substituted by: trifluoromethoxy; fluorine; chlorine; bromine; iodine; C _1-3 -alkoxy or substituted C _1-3 -alkoxy, wherein substituted C _1-3 -alkoxy means that the 2- or 3-position is replaced by amino, C _1-3 -alkylamino, 2-(C _1-3 -alk base)amino, phenyl-(C _1-3 -alkyl)amino, N-(C _1-3 -alkyl)-phenyl-(C _1-3 -alkyl)amino, pyrrolidinyl Or piperidinyl substitution; phenyl-C _1-3 -alkylamino-C _1-3 -alkyl, the substituent can also be trifluoromethyl, fluorine, chlorine, bromine, iodine, C _1-5 -Alkyl or C _1-3 -alkoxy is monosubstituted or disubstituted to the phenyl nucleus, and when disubstituted, the substituents can be the same or different; R ₆ is selected from hydrogen, C _3-10 cycloalkyl, C _1-5 alkyl, halogen; R and R are _each selected from hydrogen, C _1-5 alkyl _, C _1-4 alkoxy, aryloxy, C _1-5 alkylaryl, C _1-5 alkylaryloxy, trihalogen C _1-5 alkyl, C _3-10 cycloalkyl, mercapto, C _1-5 alkyl mercapto, aryl mercapto, sulfinyl, halogen, trihalomethyl, amino, cyano, nitro, carboxyl, oxygen Carboxyl, amide, thioamido, sulfonamide, trihalomethylsulfonyl, N-sulfinamide, S-sulfinamide, trihalomethylsulfinyl, O-carbamoyl, O -thiocarbamoyl and -NR ₁₁ R ₁₂ ; _R is selected from hydrogen, C _1-5 alkyl, C _1-5 alkoxy, aryl, aryloxy, C _1-5 alkylaryl, C _1-5 alkylaryloxy, halogen, tri Halomethyl, mercapto, hydroxyl, sulfur C _1-5 alkyl and -NR ₁₁ R ₁₂ ; R ₁₀ is selected from hydrogen, C _3-10 cycloalkyl, C _1-5 alkyl, C _2-5 alkenyl, aryl, heteroaryl, hydroxyl and heteroalicyclic; R ₁₁ and R ₁₂ are independently selected from hydrogen, C _1-5 alkyl, C _2-5 alkenyl, C _3-10 cycloalkyl, acetyl, sulfonyl, trifluoromethanesulfonyl. 2. The substituted 1H-indol-2-one compound according to claim 1, wherein R ₄ is selected from C _1-5 alkyl, halogen, sulfonamide, amino, cyano, nitro, Carboxyl and -NR ₁₁ R ₁₂ . 3. The substituted 1H-indol-2-one compound according to claim 1, wherein R ₄ and R ₅ are simultaneously or respectively selected from the following structures:

4. The substituted 1H-indol-2-one compound according to claim 1, characterized in that it comprises the following compound: (E)-3-[(3-ethoxycarbonylmethyl-2-ethoxy Carbonyl-5-methyl-1H-pyrrol-4-yl)methylene]-1H-indol-2-one; (E)-5-bromo-3-[(3-ethoxycarbonylmethyl- 2-Ethoxycarbonyl-5-methyl-1H-pyrrol-4-yl)methylene]-1H-indol-2-one; (E)-3-[(3-methoxycarbonylmethyl -2-methoxycarbonyl-5-methyl-1H-pyrrol-4-yl)methylene]-(1H)-indol-2-one; (E)-5-bromo-3-[(3 -Methoxycarbonylmethyl-2-methoxycarbonyl-5-methyl-1H-pyrrol-4-yl)methylene]-(1H)-indol-2-one; (E)-3- [(3,5 Dimethyl-2-ethoxycarbonyl-1H-pyrrol-4-yl)methylene]-(1H)-indol-2-one; (E)-5-bromo-3- [(3,5 Dimethyl-2-ethoxycarbonyl-1H-pyrrol-4-yl)methylene]-(1H)-indol-2-one; (E)-5-nitro-3 -[(3,5 Dimethyl-2-ethoxycarbonyl-1H-pyrrol-4-yl)methylene]-(1H)-indol-2-one; (E)-3-[(3 , 5 dimethyl-2-methoxycarbonyl-1H-pyrrol-4-yl) methylene]-(1H)-indol-2-one; (E)-5-nitro-3-[( 3,5 Dimethyl-2-methoxycarbonyl-1H-pyrrol-4-yl)methylene]-(1H)-indol-2-one; (E)-5-nitro-3-[ (3-Ethoxycarbonylmethyl-2-ethoxycarbonyl-5-methyl-1H-pyrrol-4-yl)methylene]-(1H)-indol-2-one; (E)- 5-carboxy-3-[(3-ethoxycarbonylmethyl-2-ethoxycarbonyl-5-methyl-1H-pyrrol-4-yl)methylene]-(1H)-indole-2 -ketone; (E)-5-aminosulfonyl-3-[(3-ethoxycarbonylmethyl-2-ethoxycarbonyl-5-methyl-1H-pyrrol-4-yl)methylene] -(1H)-indol-2-one; (E)-5-methylaminosulfonyl-3-[(3-ethoxycarbonylmethyl-2-ethoxycarbonyl-5-methyl-1H -pyrrol-4-yl)methylene]-(1H)-indol-2-one; (E)-5-(N-morpholinyl)aminosulfonyl-3-[(3-ethoxycarbonyl Methyl-2-ethoxycarbonyl-5-methyl-1H-pyrrol-4-yl)methylene]-(1H)-indol-2-one; (E)-5-(3-chlorobenzene Base) aminosulfonyl-3-[(3-ethoxycarbonylmethyl-2-ethoxycarbonyl-5-methyl-1H-pyrrol-4-yl)methylene]-(1H)-indole -2-one; (E)-5-(3-fluorophenyl)aminosulfonyl-3-[(3-ethoxycarbonylmethyl-2-ethoxycarbonyl-5-methyl-1H-pyrrole -4-yl)methylene]-(1H)-indol-2-one; (E)-5-dimethylaminosulfonyl-3-[(3-ethoxycarbonylmethyl-2-ethane Oxycarbonyl-5-methyl-1H-pyrrol-4-yl)methylene]-(1H)-indol-2-one; (E)-5-(3-fluoro-4-chlorophenyl) Aminosulfonyl-3-[(3-ethoxycarbonylmethyl-2-ethoxycarbonyl-5-methyl-1H-pyrrol-4-yl)methylene]-(1H)-indole-2 - Ketone; (E)-5-(hexahydropyridyl)aminosulfonyl-3-[(3-ethoxycarbonylmethyl-2-ethoxycarbonyl-5-methyl-1H-pyrrole-4- Base) methylene]-(1H)-indol-2-one; (E)-5-(3-bromophenyl)aminosulfonyl-3-[(3-ethoxycarbonylmethyl-2- Ethoxycarbonyl-5-methyl-1H-pyrrol-4-yl)methylene]-(1H)-indol-2-one. 5. The preparation method of the substituted 1H-indol-2-one compound according to claim 1, characterized in that under the catalysis of the base, the compound (I) is prepared by equimolar compound II and compound III in an organic solvent A condensation reaction is carried out to obtain:

6. The method for preparing substituted 1H-indol-2-one compounds according to claim 5, characterized in that the base catalyst is an organic nitrogen base or an inorganic base, and the organic solvent is a protic solvent or an aprotic solvent, The reaction temperature is 65°C-90°C; the reaction time is 3-6 hours. 7. The preparation method of substituted 1H-indol-2-one compounds according to claim 6, characterized in that the organic nitrogen base can be triethylamine, pyridine, hexahydropyridine, trimethylamine, aniline, diisopropyl amine, 1,8-diazabicyclo[5.4.1]undec-7-ene, pyrrolidine, piperidine, ammonia, alkali metal hydroxide, alkaline earth metal hydroxide, phosphate , carbonates, bicarbonates or bisulfates. 8. The method for preparing substituted 1H-indol-2-one compounds according to claim 6, characterized in that the protic solvent is water or alcohols, and the aprotic solvent is a polar or non-polar aprotic solvent , and does not contain acidic hydrogen, that is, does not contain hydrogen that can be combined with the solute, wherein the non-polar aprotic solvent can be pentane, hexane, benzene, toluene, methylene chloride or carbon tetrachloride, and the polar aprotic solvent Chloroform, tetrahydrofuran, dimethylsulfoxide or dimethylformamide can be used. 9. The method for preparing substituted 1H-indol-2-one compounds according to claim 6 or 7, characterized in that when the solvent is a protic solvent, the base catalyst is alkali metal hydroxide or alkaline earth metal hydrogen oxide. 10. The use of the substituted 1H-indol-2-one compound according to claim 1 as an epidermal growth factor receptor inhibitor in the preparation of antitumor drugs.