CN101440086B - Halogenated pyrrole substituted indolinones, and intermediate, preparation and use thereof - Google Patents

Abstract

式I 式IIThe invention discloses a class of halogenated pyrrole substituted 2-indolinones as shown in formula I; wherein R ₁ , R ₂ , R ₃ , and R ₄ are independently hydrogen, C ₁ -C ₁₂ alkyl, C ₁ -C ₁₂ alkoxy, C ₁ -C ₁₂ alkylthio, C ₁ -C ₄ haloalkyl, C ₁ -C ₄ haloalkoxy, C ₅ -C ₁₂ cycloalkyl, C ₅ -C ₁₂ heterocycle Base, C ₆ -C ₁₂ aryl, C ₆ -C ₁₂ aryloxy, hydroxyl, trihalomethyl or halogen; R ₅ and R ₆ are independently hydrogen, C ₁ -C ₁₂ alkyl or halogen; R ₇ is NR ₈ (CH ₂ ) _m R ₉ or NR ₁₀ R ₁₁ ; R ₈ is hydrogen or C ₁ -C ₁₂ alkyl; R ₉ is hydrogen, halogen, C ₁ -C ₁₂ alkyl, NR ₁₀ R ₁₁ , C(O)R ₁₂ , C(OH)R ₁₂ , C(CH ₂ OH)R ₁₂ , C ₅ -C ₁₂ heterocyclyl or C ₆ -C ₁₂ aryl; R ₁₀ , R ₁₁ , R ₁₂ alone is hydrogen, C ₁ -C ₁₂ alkyl, C ₁ -C ₁₂ alkoxy, C ₁ -C ₁₂ alkylthio, C ₁ -C ₄ haloalkyl, C ₁ -C ₄ haloalkoxy, C ₅ -C ₁₂ cycloalkyl, C ₅ -C ₁₂ heterocyclyl, C ₆ -C ₁₂ aryl or C ₆ -C ₁₂ aryloxy. The invention also discloses the preparation method of the compound and its intermediate represented by formula II, and its application in the preparation of antitumor drugs.

Formula I Formula II

Description

Halogenated pyrrole-substituted indolinones and their intermediates, preparation methods and applications

technical field

The present invention relates to a class of novel compounds and their intermediates, as well as their preparation methods and applications, in particular to halopyrrole-substituted indolinones and their intermediates, their preparation methods and applications.

Background technique

Anticancer drug research is a major topic at present. Traditional cytotoxic drugs mainly act on DNA, RNA, tubulin, etc., making these drugs poor in selectivity and high in toxicity. In recent years, a series of major advances have been made in the basic theoretical research of tumors, such as: the signal transduction in tumor cells, the regulation of cell cycle, the induction of apoptosis, the formation of tumor angiogenesis, and the role of cells and extracellular matrix have been elucidated. . As a result, some new anti-tumor drugs with specific targets on the molecules (especially key enzymes) of cell signaling pathways related to the proliferation and differentiation of tumor cells have been developed.

In recent years, the US FDA has successively approved the listing of tyrosine kinase inhibitors such as Gleevec, Iressa, and Tarceva, and achieved remarkable results. With the deepening of clinical research, some shortcomings of this class of drugs are found. Therefore, in recent years, multi-target specific drugs have emerged, hoping to inhibit tumor growth more effectively by inhibiting two or more signal transduction pathways. In recent years, the US FDA has approved sorafenib, Vandetanib, and sunitinib for marketing, and achieved good clinical results.

Contents of the invention

The technical problem to be solved by the present invention is to provide a new class of antitumor drug active substance with multi-target specificity, its intermediate and preparation method, and its application in the field of medicine.

The compound halopyrrole substituted 2-indolinone of the present invention is shown in formula I:

Formula I

Wherein, R ₁ , R ₂ , R ₃ , and R ₄ are independently: hydrogen, C ₁ -C ₁₂ alkyl, C ₁ -C ₁₂ alkoxy, C ₁ -C ₁₂ alkylthio, C ₁ -C ₄ Haloalkyl, C ₁ -C ₄ haloalkoxy, C ₅ -C ₁₂ cycloalkyl, C ₅ -C ₁₂ heterocyclyl, C ₆ -C ₁₂ aryl, C ₆ -C ₁₂ aryloxy, hydroxyl, tri Halomethyl or halogen;

R ₅ and R ₆ are independently: hydrogen, C ₁ -C ₁₂ alkyl or halogen, at least one of which is halogen;

R ₇ is: NR ₈ (CH ₂ ) _m R ₉ or NR ₁₀ R ₁₁ , m=1-10;

R ₈ is: hydrogen or C ₁ -C ₁₂ alkyl;

R ₉ is: hydrogen, halogen, C ₁ -C ₁₂ alkyl, NR ₁₀ R ₁₁ , C(O)R ₁₂ , C(OH)R ₁₂ , C(CH ₂ OH)R ₁₂ , C ₅ -C ₁₂ hetero Cyclic or C ₆ -C ₁₂ aryl;

R ₁₀ , R ₁₁ , and R ₁₂ are independently: hydrogen, C ₁ -C ₁₂ alkyl, C ₁ -C ₁₂ alkoxy, C ₁ -C ₁₂ alkylthio, C ₁ -C ₄ haloalkyl, C ₁ -C ₄ haloalkoxy, C ₅ -C ₁₂ cycloalkyl, C ₅ -C ₁₂ heterocyclyl, C ₆ -C ₁₂ aryl or C ₆ -C ₁₂ aryloxy.

Unless otherwise stated, the following terms appearing in the present specification and claims have the following meanings:

The term "alkyl" denotes branched and straight chain saturated aliphatic hydrocarbon groups having the indicated number of carbon atoms. For example, C ₁ -C ₁₀ , as defined in "C ₁ -C ₁₀ alkyl" includes those having 1, 2, 3, 4, 5, 6, 7, 8, 9 or A group of 10 carbon atoms. For example, "C ₁ -C ₁₀ alkyl" specifically includes methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, isobutyl, pentyl, hexyl, heptyl, octyl, Nonyl and decyl and so on.

The term "cycloalkyl" refers to a saturated or partially unsaturated monocyclic, polycyclic or bridged carbocyclic substituent. A ring with 3-20 carbon atoms can be represented as C _3-20 cycloalkyl; a ring with 5-15 carbon atoms can be represented as C _5-15 cycloalkyl; a ring with 3-8 carbon atoms can be Expressed as C _3-8 cycloalkyl, etc. The term includes, but is not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, 1H-indenyl, 2,3-dihydroindenyl, 1,2,3,4 -Tetrahydro-naphthyl, 5,6,7,8-tetrahydro-naphthyl, 8,9-dihydro-7H-benzocyclohepten-6-yl, 6,7,8,9-tetrahydro -5H-benzocycloheptenyl, 5,6,7,8,9,10-hexahydro-benzocyclooctenyl, fluorenyl, bicyclo[2.2.1]heptyl, bicyclo[2.2. 1] Heptenyl, bicyclo [2.2.2] octyl, bicyclo [3.1.1] heptyl, bicyclo [3.2.1] octyl, bicyclo [2.2.2] octenyl, bicyclo [ 3.2.1] Octenyl, adamantyl, octahydro-4,7-methylene-1H-indenyl, octahydro-2,5-methylene-pentalenyl and the like. Cycloalkyl groups can be attached to the center molecule via any carbon atom and can be further substituted.

The term "alkoxy" denotes a cyclic or acyclic alkyl group having the stated number of carbon atoms attached through an oxygen bridge. Thus, "alkoxy" includes the above definitions of alkyl and cycloalkyl.

The term "alkylthio" denotes a cyclic or acyclic alkyl group having the stated number of carbon atoms attached through a sulfur bridge. "Alkylthio" thus includes the above definitions of alkyl and cycloalkyl.

The term "halogen" means fluorine, chlorine, bromine, iodine, astatine.

The term "haloalkyl" means an alkyl group substituted in any position with a halogen. Thus, "haloalkyl" includes the definitions of halo and alkyl above.

The term "haloalkoxy" means an alkoxy group optionally substituted with a halogen. Thus, "haloalkoxy" includes the above definitions for halo and alkoxy.

The term "aryl" denotes any stable monocyclic or bicyclic carbocyclic ring of up to 7 atoms in each ring, at least one of which is aromatic. Examples of the aforementioned aryl units include phenyl, naphthyl, tetrahydronaphthyl, indanyl, biphenyl, phenanthrenyl, anthracenyl or acenaphthyl. In cases where the aryl substituent is a bicyclic substituent and one of the rings is non-aromatic, the attachment to the central molecule is through the aromatic ring.

The term "aryloxy" denotes an aryl group having the stated number of carbon atoms attached through an oxygen bridge. "Aryloxy" thus includes the above definition of aryl.

The term "heterocyclic group" means an aromatic or non-aromatic heterocyclic ring containing a heteroatom selected from O, N and S, and includes bicyclic groups. Thus, "heterocyclyl" includes the aforementioned heteroaryl groups as well as dihydro or tetrahydro analogs thereof. Other examples of "heterocyclyl" include, but are not limited to, the following: benzimidazolyl, benzofuranyl, benzofurazanyl, benzopyrazolyl, benzotriazolyl, benzothienyl, benzooxa Azolyl, carbazolyl, carbolinyl, cinnolinyl, furyl, imidazolyl, indolinyl, indolyl, indazolyl, isobenzofuryl, isozaindenyl, isoquinoline Base, isothiazolyl, isoxazolyl, naphthyridinyl, oxadiazolyl, oxazolyl, oxazoline, isoxazoline, oxetanyl, pyranyl, pyrazinyl, pyrazolyl, Pyridazinyl, pyridopyridyl, pyridazinyl, pyridyl, pyrimidinyl, pyrrolyl, quinazolinyl, quinolinyl, quinoxalinyl, tetrahydropyranyl, tetrazolyl, tetrazolopyridine Base, thiadiazolyl, thiazolyl, thienyl, triazolyl, azetidinyl, 1,4-dioxanyl, hexahydroazepinyl, piperazinyl, piperidinyl, pyrrolidine base, morpholinyl, thiomorpholinyl, dihydrobenzimidazolyl, dihydrobenzofuranyl, dihydrobenzothienyl, dihydrobenzoxazolyl, dihydrofuranyl, dihydroimidazolyl , Dihydroindolyl, Dihydroisoxazolyl, Dihydroisothiazolyl, Dihydrooxadiazolyl, Dihydrooxazolyl, Dihydropyrazinyl, Dihydropyrazolyl, Dihydropyridyl, Dihydropyrimidinyl, dihydropyrrolyl, dihydroquinolyl, dihydrotetrazolyl, dihydrothiadiazolyl, dihydrothiazolyl, dihydrothienyl, dihydrotriazolyl, dihydroazacyclic Butyl, methylenedioxybenzoyl, tetrahydrofuranyl and tetrahydrothienyl and their N-oxides. The heterocyclic group can be linked to the central molecule via a carbon atom or a heteroatom.

The term "hydroxy" means -OH.

The term "trihalomethyl" means a methyl group substituted with three halogen atoms. Thus, "trihalomethyl" includes the above definition of halo.

The present invention also relates to the preparation method of the halogenated pyrrole substituted 2-indolinone shown in formula I, which comprises the following steps:

(1) Combine the compound shown in formula II with HR ₇ , 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC), 1-hydroxyl-benzo-triazole (HOBt) reacts with a base; wherein, R ₇ is as previously described.

Formula II

Wherein, R ₅ and R ₆ are as previously described.

Wherein, the pH of the reaction system is preferably controlled at 9-14. The temperature of the reaction is preferably 0-40°C. The reaction time is preferably 10-30 hours. The base is preferably one or more of diethylamine, triethylamine, pyridine, sodium hydroxide and potassium hydroxide. The amount of HR ₇ , EDC, HOBt or base is preferably 0.8-1.2 times the molar amount of the compound shown in formula II. When the reactants are all solid, an aprotic solvent can be added to dissolve the reactants, and the dosage can be about 5 ml per 1 g of the reactants.

(2) Add the compound shown in formula III and a base to the mixed solution prepared in step (1) to react to prepare the halogenated pyrrole-substituted 2-indolinone shown in formula I.

Formula III

Wherein, R ₁ , R ₂ , R ₃ and R ₄ are as described above.

Wherein, the pH of the reaction system is preferably controlled between 10-14. The temperature of the reaction is preferably 50-90°C. The reaction time is preferably 2-4 hours. The amount of the compound represented by formula III is preferably 0.8-1.2 times the molar amount of the compound represented by formula II. The base is preferably one or more of diethylamine, triethylamine, pyridine, sodium hydroxide and potassium hydroxide. The amount of the base is a catalytic amount.

The present invention also relates to the intermediate compound halogenated pyrrole for the preparation of the above compound, as shown in formula II:

Formula II

Wherein, R ₅ and R ₆ are as mentioned above.

This intermediate compound can be made by following preparation method:

(1) In a mixed solvent of a protic organic solvent and water, the compound represented by formula IV is reacted with a base.

Formula IV

Wherein, the alkali is preferably sodium hydroxide and/or potassium hydroxide. The molar ratio of the compound represented by formula IV to the base is preferably 1:4˜1:9. The reaction temperature is preferably 60-80°C, more preferably reflux temperature. The reaction time is preferably 1-5 hours. The protic organic solvent is preferably one or more of methanol, ethanol, tetrahydrofuran, N,N-dimethylformamide and dimethyl sulfoxide. In the mixed solvent of the protic organic solvent and water, the ratio of the protic organic solvent and water can be 1:2-1:5; the amount of the mixed solvent of the protic organic solvent and water can be Reactant about 20ml solvent.

(2) Adjusting the pH value of the mixed solution prepared in step (1) to 1-6, the intermediate compound halogenated pyrrole represented by formula II can be prepared.

Wherein, the reagent for adjusting pH is preferably 10% hydrochloric acid.

The present invention further relates to the application of said halogenated pyrrole substituted 2-indolinone in the preparation of antitumor drugs. The halogenated pyrrole-substituted 2-indolinones of the present invention have significant antitumor activity, see the effect examples for details.

The halogenated pyrrole-substituted 2-indolinone compound of the present invention can be prepared into a pharmaceutical composition with various commonly used pharmaceutical additives (such as diluents and excipients, etc.). According to the purpose of treatment, the pharmaceutical composition can be made into various types of administration unit dosage forms, such as tablets, pills, powders, liquids, suspensions, emulsions, granules, capsules, suppositories and injections (solutions and suspensions), etc. .

For shaping pharmaceutical compositions in tablet form, any excipients known and widely used in the art may be used. For example, carriers such as lactose, white sugar, sodium chloride, glucose, urea, starch, calcium carbonate, kaolin, crystalline cellulose and silicic acid, etc.; binders such as water, ethanol, propanol, common syrup, glucose solution, starch solution, Mingjiao solution, carboxymethylcellulose, shellac, methylcellulose and potassium phosphate, polyvinylpyrrolidone, etc.; disintegrants, such as dry starch, sodium alginate, agar powder and kelp powder, sodium bicarbonate, carbonic acid Calcium, fatty acid esters of polyethylene sorbitan, sodium lauryl sulfate, monoglyceride stearate, starch and lactose, etc.; disintegration inhibitors such as white sugar, glycerol tristearate, coconut oil and hydrogenated oil; adsorption promoters, such as quaternary ammonium base and sodium lauryl sulfate, etc.; wetting agents, such as glycerin, starch, etc.; adsorbents, such as starch, lactose, kaolin, bentonite and colloidal silicic acid, etc.; and lubricants, Such as pure talc, stearate, boric acid powder and polyethylene glycol. Tablets can also be prepared as sugar-coated tablets, gelatin film-coated tablets, enteric-coated tablets, film-coated tablets, double film tablets and multi-layer tablets with usual coating materials, if desired.

To shape pharmaceutical compositions in the form of pills, any known and widely used excipients in the art may be used, for example, carriers such as lactose, starch, coconut oil, hardened vegetable oil, kaolin and talc, etc.; binders such as Gum arabic powder, tragacanth powder, gelatin and ethanol, etc.; disintegrants, such as agar and kelp powder, etc.

To shape the pharmaceutical composition in suppository form, any known and widely used excipients in the art may be used, for example, polyethylene glycol, coconut oil, higher alcohols, esters of higher alcohols, gelatin and semi-synthetic glycerides, etc. .

In order to prepare the pharmaceutical composition in the form of injection, the solution and suspension can be sterilized, and an appropriate amount of sodium chloride, glucose or glycerin is preferably added to make an injection with blood isotonic pressure. When preparing injections, any carrier commonly used in the art can also be used. For example, water, ethanol, propylene glycol, ethoxylated isostearyl alcohol, polyoxylated isostearyl alcohol, fatty acid esters of polyethylene sorbitan, and the like. In addition, common dissolving agents, buffering agents, analgesics and the like can also be added. According to needs, during the treatment of schizophrenia, coloring agents, preservatives, spices, flavoring agents, flavoring agents and other drugs may also be added.

The content of the halogenated pyrrole-substituted 2-indolinone represented by formula I of the present invention in the pharmaceutical composition can usually be 25% by mass.

In the present invention, the administration method of the pharmaceutical composition is not particularly limited. According to the patient's age, gender and other conditions and symptoms, various dosage forms of preparations can be selected for administration. For example, tablets, pills, solutions, suspensions, emulsions, granules, and capsules are administered orally; injections can be administered alone, or mixed with injection delivery fluids (such as glucose solution and amino acid solution) for intravenous injection, if available If necessary, injections can be used simply for intramuscular, intradermal, subcutaneous or intraperitoneal injection; suppositories are administered to the rectum.

In the present invention, the dosage can be appropriately selected according to the administration method, patient's age, sex and other conditions and symptoms. The usual dosage can be: about 200-800 mg active ingredient/person/day.

The reagents and raw materials used in the present invention are all commercially available.

The positive progress effect of the present invention is that: the halogenated pyrrole-substituted 2-indolinone of the present invention has remarkable antitumor activity.

Detailed ways

The present invention is further illustrated below by means of examples, but the present invention is not limited to the scope of the examples.

Example 1 2-methyl-4-chloro-5-formaldehyde-3-pyrrolecarboxylic acid

Mix ethyl 2-methyl-4-chloro-5-formyl-3-pyrrolecarboxylate (3g, 14mmol), methanol 4ml, water 9ml and sodium hydroxide 4g, heat at 60°C for 2 hours, cool, and use di Chloromethane was extracted twice, and 10% hydrochloric acid solution was added dropwise to the aqueous phase until the pH value was about 3. Brown-black precipitates were precipitated, and filtered by suction to obtain (2.1 g) 2-methyl-4-chloride with a yield of 81%. - 5-Formaldehyde-3-pyrrolecarboxylic acid solid.

¹ H NMR (CDCl ₃ ): δ 12.12 (brs, 1H); 9.66 (s, 1H); 2.6 (s, 3H).

Example 2 N-[2-(diethylamino)ethyl]-5-[(Z)-(5-fluoro-2-oxo-1,2-dihydro-3H-indole-3-ylidene ) Methyl-2-methyl-4-chloro-1H-pyrrole-3-carboxamide (26)

2-Methyl-4-chloro-5-formyl-3-pyrrole carboxylic acid (1g, 5.3mmol), EDC (5.3mmol), HOBt (5.3mmol), triethylamine (5.3mmol), N,N- Diethylethylenediamine (5.3mmol) was mixed, adjusted to pH 12, reacted at 20°C for 20h, added saturated brine 3mL, saturated sodium bicarbonate 5mL, extracted three times with 10% methanol in dichloromethane, dried and evaporated under reduced pressure. Remove the solvent, add 5-fluorooxindole (0.6g, 2.6mmol) and 0.001mol triethylamine to the remaining liquid, adjust the pH to 12, heat to 70°C, keep the reaction for 3h, cool to room temperature, and filter with suction. Wash with cold ethanol to obtain (1.03g) yield 47% N-[2-(diethylamino)ethyl]-5-[(Z)-(5-fluoro-2-oxo-1,2-dihydro -3H-indol-3-ylidene)methyl-2-methyl-4-chloro-1H-pyrrole-3-carboxamide.

¹ HNMR (DMSO-d ₆ ): δ13.85(s, 1H); 11.06(s, 1H); 6.83-7.73(m, 5H); 3.27(m, 2H); 2.45-2.54(m, 9H); 0.94(t,6H).

Example 3

Use (2S)-1-amino-3-morpholine-2-propanol instead of N,N-diethylethylenediamine to react as in Example 2.

Example 4

Use (2S)-1-amino-3-(N-methylpiperazine)-2-propanol instead of N,N-diethylethylenediamine to react as in Example 2.

Example 5 2-methyl-4-chloro-5-formaldehyde-3-pyrrolecarboxylic acid

Mix ethyl 2-methyl-4-chloro-5-formyl-3-pyrrolecarboxylate (3g, 14mmol), 4ml of ethanol, 9ml of water and potassium hydroxide (7.056g, 126mmol), and heat at 80°C for 1 hour , cooled, extracted twice with dichloromethane, 10% hydrochloric acid solution was added dropwise to the aqueous phase until the pH value was about 6, brownish-black precipitates were precipitated, filtered by suction to obtain 2-methyl-4-chloro-5-formaldehyde -3-Pyrrolecarboxylic acid solid.

Example 6 N-[2-(diethylamino)ethyl]-5-[(Z)-(5-fluoro-2-oxo-1,2-dihydro-3H-indole-3-ylidene ) Methyl-2-methyl-4-chloro-1H-pyrrole-3-carboxamide (26)

2-Methyl-4-chloro-5-formyl-3-pyrrolecarboxylic acid (1g, 5.3mmol), EDC (4.24mmol), HOBt (4.24mmol), pyridine (4.24mmol), N, N-diethyl ethylenediamine (4.24mmol), adjust the pH to 9, react at 40°C for 10h, add saturated brine 3mL, saturated sodium bicarbonate 5mL, extract three times with 10% methanol in dichloromethane, dry, and evaporate the solvent under reduced pressure , add 5-fluorooxindole (0.6g, 2.6mmol) and 0.001mol pyridine to the remaining liquid, adjust the pH to 10, heat to 50°C, and keep the reaction for 3h, cool to room temperature, suction filter, wash with cold ethanol N-[2-(diethylamino)ethyl]-5-[(Z)-(5-fluoro-2-oxo-1,2-dihydro-3H-indol-3-ylidene)methanol was obtained yl-2-methyl-4-chloro-1H-pyrrole-3-carboxamide.

Example 7 2-methyl-4-chloro-5-formaldehyde-3-pyrrolecarboxylic acid

Mix ethyl 2-methyl-4-chloro-5-formaldehyde-3-pyrrolecarboxylate (3g, 14mmol), tetrahydrofuran 4ml, water 9ml and sodium hydroxide (2.24g, 56mmol), heat at 70°C, and react 5 hours, cooled, extracted twice with dichloromethane, added dropwise 10% hydrochloric acid solution to the water phase, until the pH value was about 1, a brownish black precipitate was precipitated, filtered with suction to obtain 2-methyl-4-chloro-5-formaldehyde Base-3-pyrrolecarboxylic acid solid.

Example 8 N-[2-(diethylamino)ethyl]-5-[(Z)-(5-fluoro-2-oxo-1,2-dihydro-3H-indole-3-ylidene ) Methyl-2-methyl-4-chloro-1H-pyrrole-3-carboxamide (26)

2-Methyl-4-chloro-5-formyl-3-pyrrole carboxylic acid (1g, 5.3mmol), EDC (6.36mmol), HOBt (6.36mmol), sodium hydroxide (6.36mmol), N,N- Diethylethylenediamine (6.36mmol) was mixed, adjusted to pH 14, reacted at 0°C for 30h, added 3mL of saturated brine and 5mL of saturated sodium bicarbonate, extracted three times with 10% methanol in dichloromethane, dried, and evaporated under reduced pressure. Remove the solvent, add 5-fluorooxindole (0.6g, 2.6mmol) and 0.001mol sodium hydroxide to the remaining liquid, adjust the pH to 14, heat to 90°C, keep the reaction for 3h, cool to room temperature, and filter with suction. N-[2-(diethylamino)ethyl]-5-[(Z)-(5-fluoro-2-oxo-1,2-dihydro-3H-indole-3- subunit) methyl-2-methyl-4-chloro-1H-pyrrole-3-carboxamide.

Effect Example 1 No. 26 Oral Mice Inhibition Test——S180

1. Purpose of the test

Using mouse S180 as a model, the anti-tumor effect of No. 26 was studied.

2. Test content

Growth inhibitory effect of oral administration of No. 26 on S180 in Kunming mice.

1. Test sample

Sample: No. 26, dark brown powder.

Reference substance: SU11248,.

Reference substance: cyclophosphamide for injection (CTX), Jiangsu Hengrui Medicine Co., Ltd., batch number: 06042921.

2. Preparation method

Sample: No. 26 was dissolved with Tween-80, and then prepared to the required concentration with 0.5% CMC Na.

Reference substance: SU11248 was dissolved with Tween-80, and then prepared to the required concentration with 0.5% CMC Na. Control substance: CTX for injection, prepared to the required concentration with physiological saline during the test.

3. Animals and tumor strains

50 Kunming mice, female, weighing 18±1g, were provided by Slack Experimental Animal Co., Ltd. Certificate of Conformity: SCXK (Shanghai) 2003-0003.

Tumor strain: mouse S180 ascites type, maintained by the Pharmacology Department of Shanghai Pharmaceutical Industry Research Institute.

4. Test method

S180 ascites tumors of mice in the vigorous growth period were taken, extracted under aseptic conditions, diluted 1:5 with normal saline, and inoculated subcutaneously in the armpit of mice at 0.2 ml/only. The next day, the mice were randomly divided into 5 groups, 10 in each group.

According to the results of the preliminary experiment, three dosage groups of 160, 80, and 40 mg/kg were set for oral administration on No. 26. Administration according to body weight, oral dose 0.5ml/20g;

The dose of SU11248 in the positive control group was 80mg/kg, and the oral dose was 0.5ml/20g;

The dose of CTX in the positive control group was 30mg/kg, and the injection volume was 0.5ml/20g.

A blank control group was also set up.

Animals were dosed by body weight starting the day after inoculation.

After 9 consecutive days of administration, they were killed on the 10th day after inoculation, the tumor mass was taken and weighed, and the tumor inhibition rate was calculated according to the following formula:

5. Results

During the experiment, death occurred in the high-dose group, but no death was observed in other groups of animals. See Table 2-2 for details

Table 2-1, the inhibitory effect of No. 26 on Kunming mice S180 (X±SD)

Note: The day of inoculation is regarded as day 0;

^** indicates P value < 0.01 compared with Control.

6 Conclusion

No. 26 has an obvious inhibitory effect on the growth of S180 tumors, and its curative effect is dose-dependent. Compared with SU11248, it has better tumor inhibitory effect.

Effect Example 2 No. 26 Oral Mice Antitumor Test——Colon26

1. Purpose of the test

Taking F1 mouse colon cancer Colon26 as a model, the anti-tumor effect of No. 26 was studied.

2. Test content

Growth inhibitory effect of oral administration of No.26 on Colon26 in F1 mice.

1. Test sample

Sample: No. 26, dark brown powder.

Reference substance: SU11248,.

Reference substance: cyclophosphamide for injection (CTX), Jiangsu Hengrui Medicine Co., Ltd., batch number: 06042921.

2. Preparation method

Sample: No. 26 was dissolved with Tween-80, and then prepared to the required concentration with 0.5% CMC Na.

Reference substance: SU11248 was dissolved with Tween-80, and then prepared to the required concentration with 0.5% CMC Na. Control substance: CTX for injection, prepared to the required concentration with physiological saline during the test.

3. Animals and tumor strains

42 F1 mice, female, weighing 18±1g, provided by Slack Experimental Animal Co., Ltd., certificate of conformity: SCXK (Shanghai) 2003-0003.

Tumor strain: 1 Colon26 solid tumor mouse, which was maintained by the Pharmacology Department of Shanghai Pharmaceutical Industry Research.

4. Test method

One Colon26 solid tumor mouse in the vigorous growth stage was taken, and the tumor mass was dissected under aseptic conditions, and physiological saline was added at a ratio of 1:5, ground evenly, and 0.2ml/mouse was inoculated subcutaneously in the armpit of the mouse. The next day, the mice were randomly divided into 6 groups with 7 mice in each group.

According to the previous experimental results, the oral administration doses of No. 26 were set to three dose groups of 160, 80, and 40 mg/kg. Administration according to body weight, oral dose 0.5ml/20g;

The dose of SU11248 in the positive control group was 80mg/kg, and the oral dose was 0.5ml/20g;

The dose of CTX in the positive control group was 30mg/kg, and the injection volume was 0.5ml/20g.

A blank control group was also set up.

Animals were dosed by body weight starting the day after inoculation.

After 9 consecutive days of administration, they were killed on the 10th day after inoculation, the tumor mass was taken and weighed, and the tumor inhibition rate was calculated according to the following formula:

5. Results

Table 2-2, the inhibitory effect of No. 26 on F1 mouse Colon26 (X±SD)

Note: The day of inoculation is regarded as day 0; ^** indicates that the P value is <0.01 compared with Control.

6 Conclusion

No. 26 has obvious inhibitory effect on the growth of Colon26 tumor, and its curative effect is dose-dependent.

Effect Example 3 No. 26 nude mice tumor inhibition test NCI-H460

1. Purpose of the test

Using human lung cancer NCI-H460 BALB/CA-nu mice as a model, the anti-tumor effect of No. 26 was studied.

2. Test content

Oral administration of No. 26 to study its growth inhibitory effect on human lung cancer NCI-H460 BALB/CA-nu mouse model

1. Test sample

Sample: No. 26, dark brown powder.

Reference substance: SU11248,.

Reference substance: cyclophosphamide for injection (CTX), Jiangsu Hengrui Medicine Co., Ltd., batch number: 06042921.

2. Preparation method

Sample: No. 26 was dissolved with Tween-80, and then prepared to the required concentration with 0.5% CMC Na.

Reference substance: SU11248 was dissolved with Tween-80, and then prepared to the required concentration with 0.5% CMC Na. Control substance: CTX for injection, prepared to the required concentration with physiological saline during the test.

3. Animals and tumor strains

36 BALB/CA-nu mice, female, weighing 18±1g, were provided by Slack Experimental Animal Co., Ltd. Certificate of Conformity: SCXK (Shanghai) 2003-0003.

Tumor strain: NCI-H460 solid type of human lung cancer, maintained by the Pharmacology Department of Shanghai Pharmaceutical Industry Research Institute.

4. Test method

The human lung cancer NCI-H460 BALB/CA-nu mouse solid tumor in the vigorous growth stage was taken, and the tumor mass was dissected under sterile conditions, and the tumor mass was cut into small pieces of rice grains, and the BALB/CA-nu Mice were inoculated subcutaneously in the armpit.

After the tumor mass grew to 50-100 mm ³ , the BALB/CA-nu mice were randomly divided into 6 groups, 6 mice in each group.

According to the previous experimental results, three dosage groups of 160, 80, and 40 mg/kg were set up for the oral administration of No. 26. Administration according to body weight, oral dose 0.5ml/20g;

The dose of SU11248 in the positive control group was 80mg/kg, and the oral dose was 0.5ml/20g;

The dose of CTX in the positive control group was 30mg/kg, and the injection volume was 0.5ml/20g.

A blank control group was also set up.

Animals were grouped into groups and administered the next day, and for 9 consecutive days, the length and width of the tumor mass were measured every 3 days, and the tumor volume was calculated using the following formula:

Volume = (length × width ² )/2

On the 18th day after inoculation, the body weight was measured, the animals were sacrificed, and the tumor mass was taken and weighed.

The tumor inhibition rate was calculated according to the following formula.

5. Results

Table 2-3, the inhibitory effect of No. 26 on BALB/CA-nu mice NCI-H460 (X ± SD)

Note: The day of inoculation is regarded as day 0; ^** indicates that the P value is <0.01 compared with Control.

6 Conclusion

Experiments have shown that No. 26 has a significant inhibitory effect on the growth of NIC-H460 tumors, and its curative effect is dose-dependent.

Claims (18)

1. a class is suc as formula the 2-dihydroindole ketone of the replacement of the halogenated pyrrole shown in the I;

Formula I

Wherein, R ₁, R ₃, R ₄For: hydrogen; R ₂Be fluorine;

R ₅For: chlorine; R ₆Be methyl;

R ₇For: NR ₈(CH ₂) _mR ₉, m=2;

R ₈For: hydrogen;

R ₉For: NR ₁₀R ₁₁

R ₁₀, R ₁₁For: C ₂Alkyl.

2. the preparation method such as the described 2-dihydroindole ketone that replaces suc as formula the halogenated pyrrole shown in the I of claims is characterized in that comprising the steps:

(1) will be suc as formula the compound shown in the II and HR ₇, 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide, 1-hydroxyl-benzo-triazole and alkali reacts and gets final product;

(2) with in the prepared mixed solution of step (1), the compound and the alkali that add shown in formula III react the 2-dihydroindole ketone that can make suc as formula the replacement of the halogenated pyrrole shown in the I;

Formula II formula III

Wherein, R ₁, R ₂, R ₃, R ₄, R ₅, R ₆And R ₇As previously mentioned.

3. method as claimed in claim 2, it is characterized in that: in the step (1), the pH of the system of described reaction is controlled to be 9～14.

4. method as claimed in claim 2, it is characterized in that: in the step (1), the temperature of described reaction is 0～40 ℃.

5. method as claimed in claim 2, it is characterized in that: in the step (1), the time of described reaction is 10～30 hours.

6. method as claimed in claim 2, it is characterized in that: in step (1) and (2), described alkali is one or more in diethylamine, triethylamine, pyridine, sodium hydroxide and the potassium hydroxide.

7. method as claimed in claim 2, it is characterized in that: in the step (2), the pH of the system of described reaction is controlled to be 10～14.

8. method as claimed in claim 2, it is characterized in that: in the step (2), the temperature of described reaction is 50～90 ℃.

9. method as claimed in claim 2 is characterized in that: in step (1) and (2), and described HR ₇, 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide, 1-hydroxyl-benzo-triazole, alkali or the compound shown in formula III consumption be suc as formula the compound molar weight shown in the II 0.8～1.2 times.

10. one kind suc as formula the midbody compound halogenated pyrrole shown in the II

Formula II

Wherein, R ₅And R ₆As aforementioned.

11. the preparation method suc as formula the midbody compound halogenated pyrrole shown in the II as claimed in claim 10 is characterized in that comprising the steps:

(1) in the mixed solvent of protic organic solvent and water, will be suc as formula the compound shown in the IV and alkali reaction;

(2) the pH value to 1 of the mixed solution that makes of regulating step (1)～6 can make the midbody compound halogenated pyrrole shown in the formula II;

Formula IV

Wherein, R ₅And R ₆As aforementioned.

12. method as claimed in claim 11 is characterized in that: in the step (1), described alkali is sodium hydroxide and/or potassium hydroxide.

13. method as claimed in claim 11 is characterized in that: in the step (1), described mol ratio suc as formula the compound shown in the IV and alkali is 1: 4～1: 9.

14. method as claimed in claim 11 is characterized in that: in the step (1), the temperature of described reaction is 60～80 ℃.

15. method as claimed in claim 11 is characterized in that: in the step (1), the time of described reaction is 1～5 hour.

16. method as claimed in claim 11 is characterized in that: in the step (1), described protic organic solvent is methyl alcohol and/or ethanol.

17. the application of the 2-dihydroindole ketone that halogenated pyrrole as claimed in claim 1 replaces in the preparation antitumor drug.

18. the preparation method suc as formula the midbody compound halogenated pyrrole shown in the II as claimed in claim 10 is characterized in that comprising the steps:

In the mixed solvent of (1) one or more in tetrahydrofuran (THF), DMF and methyl-sulphoxide, and water, will be suc as formula the compound shown in the IV and alkali reaction;

(2) the pH value to 1 of the mixed solution that makes of regulating step (1)～6 can make the midbody compound halogenated pyrrole shown in the formula II;

Formula IV

Wherein, R ₅And R ₆As aforementioned.