CN115109058B - Medicine for treating gastric cancer and preparation method thereof - Google Patents

Abstract

The invention provides a medicine for treating gastric cancer and a preparation method thereof. The drug is a beta-carboline alkaloid derivative and has a structure shown in a formula (I). The beta-carboline alkaloid derivative has good gastric cancer cell proliferation inhibition activity, and therefore, can be used as a potential drug for treating gastric cancer.

Description

A kind of medicine for treating gastric cancer and preparation method thereof

technical field

The invention relates to the field of medicine, in particular, the invention relates to a medicine for treating gastric cancer and a preparation method thereof.

Background technique

Gastric cancer is one of the most common malignant tumors in the world. Although its morbidity and mortality have declined in recent years, among all malignant tumors, gastric cancer still ranks fifth in incidence and third in mortality. Surgery is the only radical method for gastric cancer, but more than 80% of gastric cancer patients are diagnosed at an advanced stage, most of which are accompanied by extensive invasion and distant metastasis, and have lost the chance of radical surgery. Treatment, including chemotherapy, radiotherapy, targeted therapy, immunotherapy, etc. Chemotherapy drugs for gastric cancer include platinum compounds, fluoropyrimidines, taxanes, anthracyclines and irinotecan, etc., all of which are effective in the treatment of gastric cancer. Combination chemotherapy can prolong the overall survival of patients more than single-agent chemotherapy.

Carbolines (carbolines) alkaloids are a large class of indole alkaloids widely distributed in nature, which belong to a kind of non-monoterpene indole alkaloids derived from tryptophan. The tricyclic system composed of pyridine includes α-carboline, β-carboline, γ-carboline, and δ-carboline, among which β-carbolines (β-carbolines) alkaloids are the most widely distributed in nature, and their structures are relatively Simple, numerous, and the most intensively studied class of carboline alkaloids. Hundreds of natural β-carboline alkaloids have been reported, which are widely distributed in nature, including plants, marine organisms, cyanobacteria, mushrooms and even animals. Methyl Harman base etc. Different β-carboline alkaloids have different biological activities, including anti-inflammatory, antibacterial, antiviral, antitumor, antimalarial, soap killing, GABAA, 5-HT2serotonin, imidazoline receptor binding activity, PDE5 receptor inhibitory activity, etc. wait.

Contents of the invention

Based on the existing β-carboline alkaloids, the present invention provides a novel β-carboline alkaloid derivative, which has good gastric cancer cell growth inhibitory activity and can be used for the treatment of gastric cancer.

The present invention provides a compound represented by formula (I), its pharmaceutically acceptable salt, stereoisomer, tautomer, solvate and prodrug:

Wherein, R _O is independently selected from hydrogen, deuterium, halogen, cyano, nitro, amino, hydroxyl, C1-6 alkyl, C1-C6 alkoxy, halogenated C1-C6 alkyl, substituted or unsubstituted C6-C14 aryl;

R ₁ and R ₂ are independently selected from hydrogen, C1-C6 alkyl, C3-C10 cycloalkyl, halogenated C1-C6 alkyl, and at least one of R ₁ and R ₂ is not hydrogen; or R ₁ , R ₂ together form -(CH ₂ ) _n -;

R ₃ is selected from C1-C6 alkyl, substituted or unsubstituted C6-C14 aryl, substituted or unsubstituted C6-C14 aryl-C1-2 alkylene;

L is selected from C1-C6 alkylene;

Het is selected from substituted or unsubstituted 5-10 membered heteroaryl;

m is selected from 1, 2, 3, 4 or 5;

n is selected from 2, 3, 4, 5 or 6.

In some preferred embodiments, R _is independently selected from hydrogen, deuterium, fluorine, chlorine, bromine, cyano, nitro, amino, hydroxyl, methyl, ethyl, isopropyl, tert-butyl, methoxy radical, ethoxy, trifluoromethyl, phenyl.

Preferably, R ₀ is selected from hydrogen.

In some preferred embodiments, R ₁ and R ₂ are independently selected from hydrogen, methyl, ethyl, isopropyl, n-propyl, tert-butyl, trifluoromethyl, cyclopropyl, cyclobutyl, Cyclopentyl, cyclohexyl, and at least one of R ₁ and R ₂ is not hydrogen; or R ₁ and R ₂ together form -CH ₂ CH ₂ -, -CH ₂ CH ₂ CH ₂ -, -CH ₂ CH _{2 CH2CH2- , -CH2CH2CH2CH2CH2- , -CH2CH2CH2CH2CH2CH2- . _ _ _ _ _ _ _}

Preferably, R ₁ is selected from hydrogen and methyl, and R ₂ is selected from methyl, isopropyl and cyclopropyl; or R ₁ and R ₂ together form -CH ₂ CH ₂ CH ₂ CH ₂ -, -CH ₂ CH _{2CH2CH2CH2- . _ _}

Preferably, R ₁ is selected from hydrogen, R ₂ is selected from isopropyl and cyclopropyl; or R ₁ is selected from methyl, R ₂ is selected from methyl; or R ₁ and R ₂ together form -CH ₂ CH ₂ CH _{2CH2CH2- . _}

In some preferred embodiments, R _is selected from methyl, ethyl, phenyl, benzyl.

Preferably, R ₃ is selected from methyl and ethyl.

_In some preferred _{embodiments ,} L is selected _{from -CH2-} , _{-CH2CH2- , -CH2CH2CH2-} , _{-CH2CH2CH2CH2- .}

Preferably, L is selected from -CH ₂ -, -CH ₂ CH ₂ -.

In some preferred embodiments, Het is selected from substituted or unsubstituted 5-7 membered heteroaryl, and the "substituted or unsubstituted" means unsubstituted, or is selected from deuterium, halogen, cyano, Substituted by nitro, amino, hydroxyl, C1-4 alkyl, C1-C4 alkoxy, halogenated C1-C4 alkyl.

Preferably, Het is selected from substituted or unsubstituted: oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, pyridyl, pyrimidinyl, the "substituted or unsubstituted" means unsubstituted, or 1-3 selected from deuterium, fluorine, chlorine, bromine, cyano, nitro, amino, hydroxyl, methyl, ethyl, isopropyl, n-propyl, methoxy, ethoxy, trifluoromethane The group substitution of the group.

More preferably, Het is selected from substituted or unsubstituted: isoxazolyl, isothiazolyl, pyridyl, pyrimidinyl, the "substituted or unsubstituted" means unsubstituted, or is selected from 1-3 A group selected from fluorine, chlorine, methyl, ethyl, isopropyl, n-propyl, trifluoromethyl is substituted.

In the present invention, the "substituted or unsubstituted" means unsubstituted or substituted by a group selected from: deuterium, halogen, cyano, nitro, amino, hydroxyl, C1-C4 alkyl, C1-C4 Alkoxy, halogenated C1-C4 alkyl.

Preferably, the "substituted or unsubstituted" means unsubstituted or substituted by a group selected from: deuterium, fluorine, chlorine, bromine, cyano, nitro, amino, hydroxyl, methyl, ethyl, Isopropyl, n-propyl, methoxy, ethoxy, trifluoromethyl.

In some preferred embodiments, the compound shown in the formula (I) has the structure shown in the following formula (II):

Wherein, R ₁ and R ₂ are independently selected from hydrogen, methyl, ethyl, isopropyl, n-propyl, tert-butyl, trifluoromethyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl , and at least one of R ₁ and R ₂ is not hydrogen; or R ₁ and R ₂ together form -CH ₂ CH ₂ -, -CH ₂ CH ₂ CH ₂ -, -CH ₂ CH ₂ CH ₂ CH ₂ -, _{-CH2CH2CH2CH2CH2- , -CH2CH2CH2CH2CH2CH2- ; _ _ _ _ _ _ _}

_R is selected from methyl, ethyl, phenyl, benzyl;

L is selected from -CH ₂ -, -CH ₂ CH ₂ -, -CH ₂ CH ₂ CH ₂ -, -CH ₂ CH ₂ CH ₂ CH ₂ -;

Het is selected from substituted or unsubstituted: oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, pyridyl, pyrimidinyl, said "substituted or unsubstituted" means unsubstituted, or replaced by 1- 3 groups selected from deuterium, fluorine, chlorine, bromine, cyano, nitro, amino, hydroxyl, methyl, ethyl, isopropyl, n-propyl, methoxy, ethoxy, trifluoromethyl group replaced.

In some preferred embodiments, R ₁ is selected from hydrogen, methyl, R ₂ is selected from methyl, isopropyl, cyclopropyl; or R ₁ and R ₂ together form -CH ₂ CH ₂ CH ₂ CH ₂ - _{, -CH2CH2CH2CH2CH2- ; _ _}

R ₃ is selected from methyl, ethyl;

L is selected from -CH ₂ -, -CH ₂ CH ₂ -;

Het is selected from substituted or unsubstituted: isoxazolyl, isothiazolyl, pyridyl, pyrimidinyl, the "substituted or unsubstituted" means unsubstituted, or is selected from 1-3 selected from fluorine , chlorine, methyl, ethyl, isopropyl, n-propyl, trifluoromethyl group substitution.

In some preferred embodiments, the compound represented by the formula (I) is selected from:

In the present invention, the term "halogen" refers to fluorine, chlorine, bromine and iodine. The term "halo" refers to substitution by fluorine, chlorine, bromine or iodine.

In the present invention, the term "heteroaryl" refers to a group containing at least 1 heteroatom selected from O, N and S, optionally containing 1-3 additional heteroatoms independently selected from O, N and S Any aromatic ring structure. The heteroaryl can be monocyclic (for example, 5-6 membered monocyclic heteroaryl), or bicyclic (for example, 9-10 membered bicyclic heteroaryl), and the heteroaryl can be connected to any N atom of the ring or C atoms in order to form a stable structure. Examples of suitable heteroaryl groups include, but are not limited to, pyrrolyl, furyl, thienyl, oxazolyl, imidazolyl, pyrazolyl, isoxazolyl, thiazolyl, isothiazolyl, triazolyl, oxadioxyl, Azolyl, thiadiazolyl, pyridyl, pyrimidinyl, pyridazinyl, pyrazinyl, pyranyl, indolizyl, indolyl, isoindolinyl, indazolyl, benzofuryl, Benzothienyl, benzimidazolyl, benzothiazolyl, purinyl, quinazinyl, quinolinyl, isoquinolyl, cinnolinyl, phthalazinyl, quinazolinyl, quinoxalinyl, naphthalene Pyridyl, pteridyl, etc.

For use in medicine, in the present invention, the term "pharmaceutically acceptable salt" includes acid addition salts obtained, for example, by reacting a solution of the compound with a pharmaceutically acceptable acid such as hydrochloric acid, sulfuric acid, phosphoric acid, transbutyl A solution of olefinic acid, maleic acid, succinic acid, acetic acid, benzoic acid, citric acid, and tartaric acid is formed by mixing. Representative pharmaceutically acceptable salts include the following: hydrochloride, dihydrochloride, hydrobromide, nitrate, bisulfate, sulfate, phosphate, hydrogen phosphate, dihydrogen phosphate, carbonic acid Salt, bicarbonate, borate, acetate, besylate, benzoate, bitartrate, camphorsulfonate, clavulanate, citrate, edetate, ethanesulfonic acid Salt, fumarate, glucoheptonate, gluconate, glutamate, hydroxynaphthoate, isothiosulfate, lactate, lactobionate, laurate, malate, cis Butenoate, fumarate, succinate, mandelate, methanesulfonate, naphthalenesulfonate, oleate, palmitate, pantothenate, polygalacturonate , Salicylate, Stearate, Tanninate, Tosylate and Valerate.

When the compounds of the present invention possess at least 1 chiral center, they may exist in enantiomeric forms accordingly. When compounds possess 2 or more chiral centers, they may exist in diastereoisomeric forms accordingly. It is to be understood that all such stereoisomers and mixtures thereof are included within the scope of the present invention.

In the present invention, the term "tautomer" refers to structural isomers of different energies that interconvert via a low energy barrier. The compounds of the present invention may exist in different tautomeric forms, all of which are included within the scope of the present invention.

In the present invention, the term "solvate" refers to an association of one or more solvent molecules with a compound of the present invention. Solvents that form solvates include, but are not limited to, water, isopropanol, ethanol, methanol, dimethyl sulfoxide, ethyl acetate, acetic acid.

In the present invention, the term "prodrug" refers to a functional derivative of the compound of the present invention, which is easily converted into the desired compound in vivo. General procedures for selecting and preparing suitable prodrug derivatives are described, for example, in "Design of Prodrugs", H. Bundgaard, ed. Elsevier, 1985.

Another aspect of the present invention is to provide a pharmaceutical composition comprising the compound represented by formula (I), its pharmaceutically acceptable salt, stereoisomer, tautomer, solvate and prodrug.

In some embodiments, the pharmaceutical composition includes a pharmaceutically acceptable excipient.

Typical pharmaceutical compositions are: powder, tablet, granule, capsule, solution, emulsion, suspension, injection, spray, aerosol, powder mist, lotion, liniment, ointment, hard Ointments, pastes, patches, etc.

The term "excipient" refers to: anti-adhesive agents, antioxidants, binders, coating agents, tableting aids, disintegrants, lubricants, emulsifiers, etc. Typical excipients include: starch, calcium carbonate, calcium phosphate, calcium stearate, croscarmellose, crospovidone, citric acid, crospovidone, cysteine, ethyl cellulose Vegan, Gelatin, Hydroxypropyl Cellulose, Hydroxypropyl Methyl Cellulose, Lactose, Magnesium Stearate, Maltitol, Mannitol, Methionine, Methyl Cellulose, Methylparaben, Polyethylene Glycol, Sodium citrate, sorbitol, stearic acid, sucrose, talc, titanium dioxide, etc.

The daily dose of the compound of the present invention can vary widely, for example 0.01-1000 mg per adult per day. For oral administration, the composition is preferably provided in the form of tablets containing 0.01, 0.05, 0.1, 0.5, 1.0, 2.5, 5.0, 10.0, 15.0, 25.0, 50.0, 100, 150, 200, 250 and 500 mg of active ingredient, for symptomatic dose adjustment for the patient to be treated. An effective amount of the drug will generally be provided at a dosage level of from about 0.01 mg/kg to about 300 mg/kg body weight per day. Preferably, the range is from about 0.5 to about 5.0 mg/kg body weight per day, most preferably, from about 1.0 to about 3.0 mg/kg body weight per day. According to the dosage regimen, the compound can be administered 1-4 times per day.

Optimal dosages to be administered can be readily determined by those skilled in the art and will vary with the particular compound used, the mode of administration, the strength of the formulation, the mode of administration and the progression of the disease condition. In addition, factors associated with the particular patient being treated, including patient age, weight, diet and frequency of administration, will result in the need to adjust dosages.

The present invention also provides a pharmaceutical combination, which is a pharmaceutical product, which may include the compound represented by the formula (I) of the present invention, its pharmaceutically acceptable salt, stereoisomer, tautomer, solvate and prodrug Drugs and other agents that may be used to treat or be helpful in the treatment of cancer.

The present invention finds that the compound of the present invention has very excellent inhibitory activity on the proliferation of tumor cells, especially gastric cancer cells, such as SGC-7901, NCI-N87, AGS, etc., which is even stronger than the positive control drug.

Therefore, the present invention also provides the use of the compound represented by formula (I) of the present invention, its pharmaceutically acceptable salt, stereoisomer, tautomer, solvate and prodrug in the preparation of medicine, said Drugs are used against tumors.

Preferably, the medicament is used for treating gastric cancer.

The present invention also provides a method for preparing the compound shown in formula (I) of the present invention, which comprises the following steps:

Compound a and compound b are condensed in glacial acetic acid to generate intermediate c, intermediate c and compound d undergo an esterification reaction to generate intermediate e, and intermediate e and intermediate f react to generate a compound shown in formula (I);

Wherein, the definitions of R ₀ -R ₃ , L, Het, and m are as described herein.

Beneficial effect

The invention provides a medicine for treating gastric cancer, and also provides a preparation method of the medicine. The medicine of the invention is a beta-carboline alkaloid derivative, which has good growth inhibitory activity of gastric cancer cells (SGC-7901, NCI-N87, AGS), and thus can be used as a potential medicine for treating gastric cancer.

Detailed ways

Hereinafter, the present invention is described in more detail to facilitate understanding of the present invention.

Those skilled in the art will recognize that the chemical reactions described herein can be used to suitably prepare many other compounds of the invention and that other methods for preparing the compounds of the invention are considered to be within the scope of the invention Inside. For example, the synthesis of those non-exemplified compounds according to the present invention can be successfully accomplished by those skilled in the art through modification methods, such as appropriate protection of interfering groups, by using other known reagents in addition to those described in the present invention, or by incorporating Reaction conditions with some routine modifications. In addition, reactions disclosed herein or known reaction conditions are also recognized to be applicable to the preparation of other compounds of this invention.

The experimental methods in the following examples are conventional methods unless otherwise specified. If no specific technique or condition is indicated in the examples, it shall be carried out according to the technique or condition described in the literature in this field, or according to the product specification.

Embodiment 1: the preparation of compound 1

Add compound 1-a (4.08g, 20mmol) in the round-bottomed flask, then add 30ml of glacial acetic acid, stir to dissolve, heat to reflux, then slowly add compound 1-b (15ml) dropwise under stirring, and reflux reaction until The reaction was complete (monitored by TLC). It was concentrated to dryness under reduced pressure, and the residue was recrystallized from methanol-cyclohexane (5:1) to obtain intermediate 1-c (4.56 g, 89%).

Add intermediate 1-c (2.56g, 10mmol) to the round bottom flask, then add 100ml of absolute ethanol, stir to dissolve, ice-bath to 0-5°C, slowly add SOCl ₂ (1.43g, 12 mmol), after the dropwise addition, the ice bath was removed and the reaction was vigorously stirred for 4 h, the solvent and excess SOCl ₂ were removed in vacuo, and the obtained intermediate 1-e was directly put into the next step.

Add 65ml of anhydrous dichloromethane to intermediate 1-e, then add 2ml of triethylamine, add compound 1-f (1.93g, 10mmol) in batches under stirring, and stir the reaction mixture at room temperature for 15h. Dilute with 50 mL of dichloromethane and wash with 120 mL of saturated aqueous sodium bicarbonate. The organic layer was separated, dried over anhydrous magnesium sulfate, the solvent was removed in vacuo, and the residue was chromatographed on silica gel eluting with ethyl acetate:petroleum ether with a volume gradient of 0:100 to 30:80 to give a white solid Compound 1 (3.70 g, 84%).

HR-ESI-MS m/z 441.1580 [M+H] ⁺ (theoretical 441.1597).

Elemental analysis: C ₂₂ H ₂₄ N ₄ O ₄ S Found: C, 59.92; H, 5.47; N, 12.70; O, 14.43; S, 7.38 (Theoretical: C, 59.98; H, 5.49; N, 12.72; O, 14.53; S, 7.28).

¹ H NMR (400MHz, DMSO-d ₆ )δ9.61(s, 1H), 8.63(d, J=7.0Hz, 2H), 7.51(d, J=7.1Hz, 1H), 7.35(d, J= 7.2Hz, 1H), 7.19(t, J=7.0Hz, 1H), 7.14–7.02(m, 2H), 5.19(d, J=6.8Hz, 1H), 4.85–4.72(m, 3H), 4.12( q,J=6.4Hz,2H),3.19–3.05(m,2H),2.22(dp,J=6.8,5.6Hz,1H),1.23(t,J=6.4Hz,3H),0.63–0.42(m ,4H).

Embodiment 2: the preparation of compound 2

Add compound 1-a (4.08g, 20mmol) in the round bottom flask, then add 30ml of glacial acetic acid, stir to dissolve, heat to reflux, then slowly add compound 2-b (20ml) dropwise under stirring, reflux reaction until The reaction was complete (monitored by TLC). It was concentrated to dryness under reduced pressure, and the residue was recrystallized from methanol to obtain intermediate 2-c (4.4 g, 77%).

Add intermediate 2-c (2.84g, 10mmol) to the round bottom flask, then add 120ml of absolute ethanol, stir to dissolve, ice-bath to 0-5°C, slowly add SOCl ₂ (1.43g, 12 mmol), after the dropwise addition, the ice bath was removed and the reaction was vigorously stirred for 4 h, the solvent and excess SOCl ₂ were removed in vacuo, and the obtained intermediate 2-e was directly put into the next step.

Add 80ml of anhydrous dichloromethane to intermediate 2-e, then add 2ml of triethylamine, add compound 1-f (1.93g, 10mmol) in batches under stirring, and stir the reaction mixture at room temperature for 18h. Dilute with 60 mL of dichloromethane and wash with 120 mL of saturated aqueous sodium bicarbonate. The organic layer was separated, dried over anhydrous magnesium sulfate, the solvent was removed in vacuo, and the residue was chromatographed on silica gel eluting with ethyl acetate:petroleum ether with a volume gradient of 0:100 to 35:75 to give a light yellow Solid compound 2 (3.75 g, 80%).

HR-ESI-MS m/z 469.1922 [M+H] ⁺ (theoretical 469.1910).

Elemental analysis: C ₂₄ H ₂₈ N ₄ O ₄ S Found: C, 61.58; H, 6.05; N, 11.99; O, 13.76; S, 6.69 (Theoretical: C, 61.52; H, 6.02; N, 11.96; O, 13.66; S, 6.84).

¹ H NMR (400MHz, DMSO-d ₆ )δ10.25(s, 1H), 8.63(d, J=7.0Hz, 2H), 7.47–7.38(m, 2H), 7.19(t, J=7.1Hz, 1H), 7.15–7.03(m, 2H), 4.76(s, 2H), 4.71(t, J＝8.6Hz, 1H), 4.12(q, J＝6.4Hz, 2H), 3.17–3.02(m, 2H ), 2.20 (m, 2H), 2.02–1.91 (m, 2H), 1.88–1.74 (m, 4H), 1.55–1.38 (m, 2H), 1.23 (t, J=6.4Hz, 3H).

Embodiment 3: the preparation of compound 3

Following the same method as in Example 1, except that 1-b was replaced by 3-b and 1-f was replaced by 3-f, Compound 3 was obtained.

HR-ESI-MS m/z 463.1225 [M+H] ⁺ (theoretical 463.1207).

Elemental analysis: C ₂₁ H ₂₃ ClN ₄ O ₄ S Measured value: C, 54.39; H, 5.05; Cl, 7.60; N, 12.14; O, 13.93; S, 6.90 (theoretical value: C, 54.48; H, 5.01; Cl, 7.66; N, 12.10; O, 13.82; S, 6.93).

¹ H NMR (400MHz,DMSO-d ₆ )δ10.47(s,1H),8.07(s,2H),7.49–7.40(m,2H),7.17–7.05(m,2H),4.80(s,2H ),4.73(t,J=8.6Hz,1H),4.12(q,J=6.4Hz,2H),3.17–3.02(m,2H),1.78(s,3H),1.73(s,3H),1.23 (t, J=6.4Hz, 3H).

Embodiment 4: the preparation of compound 4

Following the same method as in Example 1, except that 1-b was replaced by 3-b and 1-f was replaced by 4-f, compound 4 was obtained.

HR-ESI-MS m/z 463.1225 [M+H] ⁺ (theoretical 442.1801).

Elemental analysis: C ₂₃ H ₂₇ N ₃ O ₄ S Measured value: C, 62.63; H, 6.19; N, 9.54; O, 14.38; S, 7.20 (theoretical value: C, 62.56; H, 6.16; N, 9.52; O, 14.49; S, 7.26).

¹ H NMR (400MHz,DMSO-d ₆ )δ10.47(s,1H),8.43-8.41(m,2H),7.54(d,J=7.7,1H),7.49–7.40(m,2H),7.29 –7.22(m,1H),7.17–7.05(m,2H),4.72(t,J=8.6Hz,1H),4.12(q,J=6.4Hz,2H),3.60–3.42(m,2H), 3.16–2.94 (m, 4H), 1.78 (s, 3H), 1.73 (s, 3H), 1.23 (t, J=6.4Hz, 3H).

Embodiment 5: the preparation of compound 5

Following the same method as in Example 1, except that 1-b was replaced by 2-b and 1-f was replaced by 5-f, compound 5 was obtained.

HR-ESI-MS m/z 458.1770 [M+H] ⁺ (theoretical 458.1750).

Elemental analysis: C ₂₃ H ₂₇ N ₃ O ₅ S Found: C, 60.35; H, 5.96; N, 9.23; O, 17.44; S, 7.06 (Theoretical: C, 60.38; H, 5.95; N, 9.18; O, 17.48; S, 7.01).

¹ H NMR (400MHz,DMSO-d ₆ )δ10.25(s,1H),8.61(d,J＝7.2Hz,1H),7.47–7.38(m,2H),7.15–7.03(m,2H), 6.53(d, J=7.2Hz, 1H), 4.76(t, J=8.6Hz, 1H), 4.65–4.56(m, 2H), 4.12(q, J=6.4Hz, 2H), 3.17–3.02(m ,2H), 2.20(m,2H), 2.02–1.91(m,2H), 1.88–1.74(m,4H), 1.56–1.37(m,2H), 1.23(t,J=6.4Hz,3H).

Embodiment 6: the preparation of compound 6

Following the same method as in Example 1, except that 1-b was replaced by 6-b and 1-f was replaced by 6-f, compound 6 was obtained.

HR-ESI-MS m/z 491.1535 [M+H] ⁺ (theoretical 491.1520).

Elemental analysis: C ₂₃ H ₂₇ ClN ₄ O ₄ S Found: C, 56.28; H, 5.55; Cl, 7.17; N, 11.40; O, 13.09; S, 6.43 (Theoretical: C, 56.26; H, 5.54; Cl, 7.22; N, 11.41; O, 13.03; S, 6.53).

¹ H NMR (400MHz, DMSO-d ₆ )δ9.29(s, 1H), 7.48(d, J=7.2Hz, 1H), 7.31(d, J=7.2Hz, 1H), 7.12–7.00(m, 2H), 6.84(s, 1H), 5.26(d, J=7.4Hz, 1H), 4.79(s, 2H), 4.74(t, J=8.6Hz, 1H), 4.12(q, J=6.4Hz, 2H), 3.19–3.05(m, 2H), 2.38(s, 3H), 2.13(septet, J=6.6Hz, 1H), 1.23(d, J=8.6Hz, 3H), 0.97(d, J=6.6 Hz, 6H).

Drug efficacy example: MTT method to test the activity of inhibiting gastric cancer cells

SGC-7901, NCI-N87, and AGS cells (from the cell bank of Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences) were returned to liquid and passaged according to conventional tumor cell culture methods, and cell lines in logarithmic growth phase were collected and seeded in 96-well culture plates respectively. , the number of cells per well was 1.0×10 ⁵ /100 μL, and then placed in a 5% CO ₂ incubator for culture, the medium was removed the next day, and 100 μL of drugs with different concentrations were added (the drug concentration was double-diluted), and 3 cells were set for each drug. Parallel sample, no drug was added to the negative control group, 5-fluorouracil (5-FU) was used as the positive control, after 48 hours, 10 μL of MTT was added to each well, and the culture was continued for 4 hours, and then 100 μL of DMSO was added to each well to terminate the reaction, left at room temperature for 1 hour, and enzyme-labeled The absorbance OD value of each well at 570nm was detected by an instrument, and the cell growth inhibition rate was calculated (growth inhibition rate (%)=[(OD _control -OD _experiment )/(OD _control -OD _blank )]×100%). The obtained cell growth inhibition rate was fitted by SPSS 22.0, and the half inhibitory rate IC ₅₀ of different drugs on SGC-7901, NCI-N87, AGS cell growth was obtained. The results are shown in Table 1 below:

Table 1: Proliferation inhibitory activity of gastric cancer cells

As shown in Table 1, the compound of the present invention has good inhibitory activity on the proliferation of gastric cancer cells (SGC- ₇₉₀₁ , NCI-N87, AGS), especially the compound ₂ and 5 has better inhibitory activity.

The above is only a preferred embodiment of the present invention, it should be pointed out that for those of ordinary skill in the art, without departing from the method of the present invention, some improvements and supplements can also be made, and these improvements and supplements should also be considered Be the protection scope of the present invention.

Claims (8)

1. A compound of formula (I), pharmaceutically acceptable salts, stereoisomers and tautomers thereof:

wherein R is ₀ Independently selected from hydrogen;

R ₁ 、R ₂ independently selected from hydrogen, methyl, ethyl, isopropyl, n-propyl, trifluoromethyl, cyclopropyl, cyclobutyl, and R ₁ 、R ₂ At least one of which is not hydrogen; or R ₁ 、R ₂ Together form-CH ₂ CH ₂ CH ₂ CH ₂ -、-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ -；

R ₃ Selected from methyl, ethyl;

l is selected from-CH ₂ -、-CH ₂ CH ₂ -；

Het is selected from substituted or unsubstituted: isoxazolyl, isothiazolyl, pyridyl, pyrimidinyl, said "substituted or unsubstituted" meaning unsubstituted or substituted with 1 to 3 groups selected from deuterium, fluorine, chlorine, bromine, methyl, ethyl, isopropyl, n-propyl, methoxy, ethoxy, trifluoromethyl;

m is selected from 1, 2, 3 or 4.

2. The compound of formula (I), pharmaceutically acceptable salts, stereoisomers and tautomers thereof according to claim 1, wherein R is ₁ Selected from hydrogen, methyl, R ₂ Selected from methyl, isopropyl, cyclopropyl.

3. The compound of formula (I), pharmaceutically acceptable salts, stereoisomers and tautomers thereof according to claim 1, wherein said compound of formula (I) has the structure represented by the following formula (II):

wherein R is ₁ 、R ₂ Independently selected from hydrogen, methyl, isopropyl, cyclopropyl, and R ₁ 、R ₂ At least one of which is not hydrogen; or R ₁ 、R ₂ Together form-CH ₂ CH ₂ CH ₂ CH ₂ -、-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ -；

R ₃ Selected from methyl, ethyl;

l is selected from-CH ₂ -、-CH ₂ CH ₂ -；

Het is selected from substituted or unsubstituted: isoxazolyl, pyridyl, pyrimidinyl, said "substituted or unsubstituted" meaning unsubstituted or substituted by 1 to 3 groups selected from deuterium, fluoro, chloro, bromo, methyl, ethyl, trifluoromethyl.

4. The compound of formula (I), pharmaceutically acceptable salts, stereoisomers and tautomers thereof according to claim 1, wherein said compound of formula (I) is selected from the group consisting of:

5. a pharmaceutical composition comprising a compound according to any one of claims 1-4, pharmaceutically acceptable salts, stereoisomers and tautomers thereof.

6. Use of a compound according to any one of claims 1-4, pharmaceutically acceptable salts, stereoisomers and tautomers thereof, for the preparation of a medicament for anti-tumor.

7. The use according to claim 6, wherein the medicament is for the treatment of gastric cancer.

8. A process for the preparation of a compound of formula (I) as claimed in claim 1, comprising the steps of:

condensing the compound a and the compound b in glacial acetic acid to generate an intermediate c, carrying out esterification reaction on the intermediate c and the compound d to generate an intermediate e, and reacting the intermediate e with the intermediate f to generate the compound shown in the formula (I);

wherein R is ₀ -R ₃ L, het, m are as defined in claim 1.