CN118084903A - A β-carboline hydrogen sulfide donor derivative and its preparation method and application - Google Patents

Abstract

The invention relates to the technical field of medicines, in particular to a beta-carboline hydrogen sulfide donor derivative, a preparation method and application thereof. The synthesis method of the beta-carboline hydrogen sulfide donor derivative is simple, firstly hydroxylamine oxysulfonic acid is used for reacting with substituted thiobenzoic acid to obtain an intermediate, then the intermediate is condensed with different beta-carboline compounds to obtain the beta-carboline hydrogen sulfide donor with double effects, the compounds have remarkable activity of resisting myocardial cell oxidative damage, beta-carboline with myocardial protection and hydrogen sulfide gas with vascular protection can be released under the action of mercaptan, damaged cardiac muscle can be protected simultaneously from two aspects, the influence on normal cells is small, and a new thought and method are provided for treating ischemic heart diseases.

Description

A β-carboline hydrogen sulfide donor derivative and its preparation method and application

Technical Field

The invention relates to the field of medical technology, in particular to a β-carboline hydrogen sulfide donor derivative and a preparation method and application thereof.

Background Art

Studies have shown that cardiovascular diseases such as myocardial infarction and coronary heart disease can cause myocardial ischemia and lead to myocardial necrosis. Timely restoration of blood flow is the main treatment for improving myocardial ischemia, but reperfusion itself will further aggravate myocardial cell damage, induce apoptosis, increase infarct area, lead to arrhythmia and even death. Therefore, myocardial ischemia/reperfusion injury (MIRI) has become an important cause of death caused by myocardial ischemia.

Hydrogen sulfide (H ₂ S), a new gas signal molecule, plays a wide range of physiological and pathological roles in the body, such as vasodilation, lowering blood pressure, inhibiting oxidative stress, regulating inflammation and vascular protection. H ₂ S can dilate blood vessels, lower blood pressure, and promote the proliferation of endothelial cells. Studies have shown that H ₂ S can significantly improve a variety of cardiovascular diseases such as arrhythmias, cardiac hypertrophy, myocardial fibrosis and heart failure. In vitro studies have shown that H ₂ S donors can reduce myocardial cell death caused by hypoxia/reoxygenation injury; in MIRI, H ₂ S can protect myocardial cells and significantly reduce the area of infarction. With the continuous deepening of research on H ₂ S as an important endogenous gas signal molecule in the human body, the research on H ₂ S donors has also made great progress, but there is still no H ₂ S donor drug on the market. NaHS and Na ₂ S are the first typical H ₂ S donors, which can release a large amount of H ₂ S in a short time, but this type of inorganic donor has a pungent smell, is unstable in nature, and has a short half-life, making it difficult to effectively treat diseases. The chemical fragments of existing organic hydrogen sulfide donors after releasing H ₂ S have almost no myocardial protective effect, resulting in low efficiency of chemical structure utilization of donor molecules. In order to achieve the micromolar concentration of H ₂ S to exert biological effects, millimolar organic donors are required, which will cause problems such as toxicity, solubility, and interaction with other components of the organism. Therefore, how to improve the efficiency of chemical structure utilization of H ₂ S donors and reduce the dosage is an urgent problem to be solved in the design of new H ₂ S donors.

Carberine is a heterocyclic compound in which a pyridine ring is fused with the pyrrole ring of indole. Studies have shown that β-carberine has significant myocardial protective activity in vitro and in vivo, but its vascular protective activity has not been reported. Patent No. 202010151142.3 reports that this type of compound is stable in solution and has significant protective activity against myocardial hypoxia-reoxygenation injury, especially hypoxia-reoxygenation injury caused by myocardial ischemia-reperfusion injury. Coupling β-carberine, which has myocardial protective activity, with hydrogen sulfide, which has cardiovascular protective effects, can simultaneously exert dual myocardial and cardiovascular protective effects, reduce the dosage, and improve the efficiency of the use of the chemical structure, providing a new idea for the development of drugs for ischemic heart disease.

Summary of the invention

In view of the above technical problems, the technical problem to be solved by the present invention is to provide a β-carboline hydrogen sulfide donor derivative and its preparation method and application

This is achieved specifically through the following technical solutions:

The first object of the present invention is to provide a β-carboline hydrogen sulfide donor derivative, the structural formula of the derivative is as follows:

Among them, _R1 is a proton, an alkyl group, a phenyl group, or an aralkyl group; _R2 is a proton, an alkyl group, a benzyl group, or a substituted benzyl group; _R3 is a proton, an alkyl group, a methoxy group, a hydroxyl group, a nitro group, an amino group, or a halogen group; and _R4 is a proton, an alkyl group, a methoxy group, a hydroxyl group, a nitro group, an amino group, or a halogen group.

Preferably, the alkyl group in structure (1) refers to a straight-chain or branched saturated aliphatic group, such as methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, tert-butyl, n-pentyl, and n-hexyl.

Preferably, _R1 is a proton, a methyl group, or an ethyl group; _R2 is a proton, a methyl group, a benzyl group, an o-fluorobenzyl group, a p-fluorobenzyl group, an o-chlorobenzyl group, a p-chlorobenzyl group, a p-trifluoromethylbenzyl group, an o-trifluoromethylbenzyl group, an o-methylbenzyl group, a p-methylbenzyl group, an o-methoxybenzyl group, or a p-methoxybenzyl group; _R3 is a proton, a methyl group, a methoxy group, or a hydroxyl group; and _R4 is a proton, a methyl group, a methoxy group, or a hydroxyl group.

Preferably, the β-carboline hydrogen sulfide donor derivative is any one of the following compounds:

Compound 3: N-(Benzoylthio)-1-ethyl-9H-pyrido[3,4-b]indole-3-carboxamide;

Compound 4: N-(Benzoylthio)-1-ethyl-9-benzyl-pyridin[3,4-b]indole-3-carboxamide;

Compound 5: N-(Benzoylthio)-1-ethyl-9-(4-fluorobenzyl)-pyrido[3,4-b]indole-3-carboxamide;

Compound 6: N-(Benzoylthio)-1-ethyl-9-(4-chlorobenzyl)-pyridin[3,4-b]indole-3-carboxamide;

Compound 7: N-(Benzoylthio)-1-ethyl-9-(4-methoxybenzyl)-pyridin[3,4-b]indole-3-carboxamide;

Compound 8: N-(Benzoylthio)-1-methyl-9-benzyl-pyridin[3,4-b]indole-3-carboxamide;

Compound 9: N-(Benzoylthio)-1-methyl-9-(4-fluorobenzyl)-pyridin[3,4-b]indole-3-carboxamide;

Compound 10: N-(Benzoylthio)-1-methyl-9-(4-chlorobenzyl)-pyridin[3,4-b]indole-3-carboxamide;

Compound 11: N-(Benzoylthio)-1-ethyl-9-methyl-pyrido[3,4-b]indole-3-carboxamide;

Compound 12: N-(Benzoylthio)-1-ethyl-9-(4-methylbenzyl)-pyrido[3,4-b]indole-3-carboxamide;

Compound 13: N-(Benzoylthio)-1-ethyl-9-(2-methylbenzyl)-pyrido[3,4-b]indole-3-carboxamide;

Compound 14: N-(Benzoylthio)-1-ethyl-9-(2-methoxybenzyl)-pyrido[3,4-b]indole-3-carboxamide;

Compound 15: N-(Benzoylthio)-1-ethyl-9-(4-trifluoromethylbenzyl)-pyrido[3,4-b]indole-3-carboxamide;

Compound 16: N-(Benzoylthio)-1-ethyl-9-(2-trifluoromethylbenzyl)-pyrido[3,4-b]indole-3-carboxamide;

Compound 17: N-(Benzoylthio)-1-ethyl-9-(2-chlorobenzyl)-pyrido[3,4-b]indole-3-carboxamide;

Compound 18: N-(Benzoylthio)-1-ethyl-9-(4-fluorobenzyl)-pyrido[3,4-b]indole-3-carboxamide;

Compound 19: 1-ethyl-6-hydroxy-9-(4-fluorobenzyl)-N-(2-methoxybenzoylthio)-pyrido[3,4-b]indole-3-carboxamide;

Compound 20: 1-ethyl-6-methyl-9-(4-fluorobenzyl)-N-(2-fluorobenzoylthio)-pyrido[3,4-b]indole-3-carboxamide.

The second object of the present invention is to provide a method for preparing the aforementioned β-carboline hydrogen sulfide donor derivative, comprising the following steps:

Step 1: Weigh KOH and dissolve it in water, stir at room temperature, add compound 1 and hydroxylamine sulfonic acid, stir to react, extract with dichloromethane, and spin dry to obtain intermediate 2;

Step 2: Weigh intermediate 2 and dissolve it in dry DMF, then add compound 2, HATU and DIEA, stir the reaction at room temperature, add pure water, extract with ethyl acetate, combine the extracts and backwash with saturated brine, spin dry the organic layer to obtain a crude product, and purify it by normal phase silica gel column chromatography to obtain compound 3-20.

The compound 1 is a substituted thiobenzoic acid, and its structural formula is: Wherein, R ₄ is a proton, an alkyl group, a methoxy group, a hydroxyl group, a nitro group, an amino group, or a halogen group;

The compound 2 is 1-ethyl-9H-pyridine [3,4-b] indole-3-carboxylic acid, 1-ethyl-9-benzyl-pyridine [3,4-b] indole-3-carboxylic acid, 1-ethyl-9-(4-fluorobenzyl)-pyrido [3,4-b] indole-3-carboxylic acid, 1-ethyl-9-(4-chlorobenzyl)-pyridine [3,4-b] indole-3-carboxylic acid, 1-ethyl-9-(4-methoxybenzyl)-pyridine [3,4 -b] indole-3-carboxylic acid, 1-methyl-9-benzyl-pyrido[3,4-b] indole-3-carboxylic acid, 1-methyl-9-(4-fluorobenzyl)-pyrido[3,4-b] indole-3-carboxylic acid, 1-methyl-9-(4-chlorobenzyl)-pyrido[3,4-b] indole-3-carboxylic acid, 1-ethyl-9-methyl-pyrido[3,4-b] indole-3-carboxylic acid, 1-ethyl-9-(4-methylbenzyl)-pyrido[3,4-b] indole-3-carboxylic acid [3,4-b]indole-3-carboxylic acid, 1-ethyl-9-(2-methylbenzyl)-pyrido[3,4-b]indole-3-carboxylic acid, 1-ethyl-9-(2-methoxybenzyl)-pyrido[3,4-b]indole-3-carboxylic acid, 1-ethyl-9-(4-trifluoromethylbenzyl)-pyrido[3,4-b]indole-3-carboxylic acid, 1-ethyl-9-(2-trifluoromethylbenzyl)-pyrido[3,4-b]indole- 3-carboxylic acid, 1-ethyl-9-(2-chlorobenzyl)-pyrido[3,4-b]indole-3-carboxylic acid, 1-ethyl-9-(4-fluorobenzyl)-pyrido[3,4-b]indole-3-carboxylic acid, 1-ethyl-6-hydroxy-9-(4-fluorobenzyl)-pyrido[3,4-b]indole-3-carboxylic acid, 1-ethyl-6-methyl-9-(4-fluorobenzyl)-pyrido[3,4-b]indole-3-carboxylic acid.

Furthermore, the present invention provides a synthetic route for the β-carboline hydrogen sulfide donor compound, which is as follows:

Furthermore, the present invention also provides a synthetic route and a preparation method of compound 3, and the synthetic route is as follows:

The preparation method steps are as follows:

Step 1: Weigh KOH and dissolve it in water, stir at room temperature, add hydroxylamine sulfonic acid and thiobenzoic acid, stir to react, extract with dichloromethane, and spin dry to obtain intermediate 2;

Step 2: Weigh the intermediate 2 and dissolve it in dry DMF, add 1-ethyl-9H-pyridine [3,4-b] indole-3-carboxylic acid, HATU and DIEA, stir the reaction at room temperature, add pure water, extract with ethyl acetate, combine the extracts and backwash with saturated brine, spin dry the organic layer to obtain a crude product, and purify it by normal phase silica gel column chromatography to obtain compound 3.

Compound 4: Preparation of N-(benzoylthio)-1-ethyl-9-benzyl-pyridin[3,4-b]indole-3-carboxamide

Referring to the preparation of compound 3, 1-ethyl-9H-pyrido[3,4-b]indole-3-carboxylic acid was replaced with 1-ethyl-9-benzyl-pyrido[3,4-b]indole-3-carboxylic acid;

Compound 5: Preparation of N-(benzoylthio)-1-ethyl-9-(4-fluorobenzyl)-pyrido[3,4-b]indole-3-carboxamide

Referring to the preparation of compound 3, 1-ethyl-9H-pyrido[3,4-b]indole-3-carboxylic acid was replaced with 1-ethyl-9-(4-fluorobenzyl)-pyrido[3,4-b]indole-3-carboxylic acid;

Compound 6: Preparation of N-(benzoylthio)-1-ethyl-9-(4-chlorobenzyl)-pyridin[3,4-b]indole-3-carboxamide

Referring to the preparation of compound 3, 1-ethyl-9H-pyrido[3,4-b]indole-3-carboxylic acid was replaced with 1-ethyl-9-(4-chlorobenzyl)-pyrido[3,4-b]indole-3-carboxyl;

Compound 7: Preparation of N-(benzoylthio)-1-ethyl-9-(4-methoxybenzyl)-pyridin[3,4-b]indole-3-carboxamide

Referring to the preparation of compound 3, 1-ethyl-9H-pyrido[3,4-b]indole-3-carboxylic acid was replaced with 1-ethyl-9-(4-methoxybenzyl)-pyrido[3,4-b]indole-3-carboxylic acid;

Compound 8 - Preparation of N-(benzoylthio)-1-methyl-9-benzyl-pyridinyl[3,4-b]indole-3-carboxamide

Referring to the preparation of compound 3, 1-ethyl-9H-pyrido[3,4-b]indole-3-carboxylic acid was replaced with 1-methyl-9-benzyl-pyrido[3,4-b]indole-3-carboxylic acid;

Compound 9: Preparation of N-(benzoylthio)-1-methyl-9-(4-fluorobenzyl)-pyridin[3,4-b]indole-3-carboxamide

Referring to the preparation of compound 3, 1-ethyl-9H-pyrido[3,4-b]indole-3-carboxylic acid was replaced with 1-methyl-9-(4-fluorobenzyl)-pyrido[3,4-b]indole-3-carboxylic acid;

Compound 10: Preparation of N-(benzoylthio)-1-methyl-9-(4-chlorobenzyl)-pyridinyl[3,4-b]indole-3-carboxamide

Referring to the preparation of compound 3, 1-ethyl-9H-pyrido[3,4-b]indole-3-carboxylic acid was replaced with 1-methyl-9-(4-chlorobenzyl)-pyrido[3,4-b]indole-3-carboxylic acid;

Compound 11——Preparation of N-(benzoylthio)-1-ethyl-9-methyl-pyrido[3,4-b]indole-3-carboxamide

Referring to the preparation of compound 3, 1-ethyl-9H-pyrido[3,4-b]indole-3-carboxylic acid was replaced with 1-ethyl-9-methyl-pyrido[3,4-b]indole-3-carboxylic acid.

Compound 12: Preparation of N-(benzoylthio)-1-ethyl-9-(4-methylbenzyl)-pyrido[3,4-b]indole-3-carboxamide

Referring to the preparation of compound 3, 1-ethyl-9H-pyrido[3,4-b]indole-3-carboxylic acid was replaced with 1-ethyl-9-(4-methylbenzyl)-pyrido[3,4-b]indole-3-carboxylic acid.

Compound 13: Preparation of N-(benzoylthio)-1-ethyl-9-(2-methylbenzyl)-pyrido[3,4-b]indole-3-carboxamide

Referring to the preparation of compound 3, 1-ethyl-9H-pyrido[3,4-b]indole-3-carboxylic acid was replaced with 1-ethyl-9-(2-methylbenzyl)-pyrido[3,4-b]indole-3-carboxylic acid.

Compound 14: Preparation of N-(benzoylthio)-1-ethyl-9-(2-methoxybenzyl)-pyrido[3,4-b]indole-3-carboxamide

Referring to the preparation of compound 3, 1-ethyl-9H-pyrido[3,4-b]indole-3-carboxylic acid was replaced with 1-ethyl-9-(2-methoxybenzyl)-pyrido[3,4-b]indole-3-carboxylic acid.

Compound 15: Preparation of N-(benzoylthio)-1-ethyl-9-(4-trifluoromethylbenzyl)-pyrido[3,4-b]indole-3-carboxamide

Referring to the preparation of compound 3, 1-ethyl-9H-pyrido[3,4-b]indole-3-carboxylic acid was replaced with 1-ethyl-9-(4-trifluoromethylbenzyl)-pyrido[3,4-b]indole-3-carboxylic acid.

Compound 16: Preparation of N-(benzoylthio)-1-ethyl-9-(2-trifluoromethylbenzyl)-pyrido[3,4-b]indole-3-carboxamide

Referring to the preparation of compound 3, 1-ethyl-9H-pyrido[3,4-b]indole-3-carboxylic acid was replaced with 1-ethyl-9-(2-trifluoromethylbenzyl)-pyrido[3,4-b]indole-3-carboxylic acid.

Compound 17: Preparation of N-(benzoylthio)-1-ethyl-9-(2-chlorobenzyl)-pyrido[3,4-b]indole-3-carboxamide

Referring to the preparation of compound 3, 1-ethyl-9H-pyrido[3,4-b]indole-3-carboxylic acid was replaced with 1-ethyl-9-(2-chlorobenzyl)-pyrido[3,4-b]indole-3-carboxylic acid.

Compound 18: Preparation of N-(benzoylthio)-1-ethyl-9-(4-fluorobenzyl)-pyrido[3,4-b]indole-3-carboxamide

Referring to the preparation of compound 3, 1-ethyl-9H-pyrido[3,4-b]indole-3-carboxylic acid was replaced with 1-ethyl-9-(4-fluorobenzyl)-pyrido[3,4-b]indole-3-carboxylic acid.

Compound 19——Preparation of 1-ethyl-6-hydroxy-9-(4-fluorobenzyl)-N-(2-methoxybenzoylthio)-pyrido[3,4-b]indole-3-carboxamide

Referring to the preparation of compound 3, thiobenzoic acid was replaced by o-methoxythiobenzoic acid, and 1-ethyl-9H-pyrido[3,4-b]indole-3-carboxylic acid was replaced by 1-ethyl-6-hydroxy-9-(4-fluorobenzyl)-pyrido[3,4-b]indole-3-carboxylic acid.

Compound 20: Preparation of 1-ethyl-6-methyl-9-(4-fluorobenzyl)-N-(2-fluorobenzoylthio)-pyrido[3,4-b]indole-3-carboxamide

Referring to the preparation of compound 3, thiobenzoic acid was replaced by o-fluorothiobenzoic acid, and 1-ethyl-9H-pyrido[3,4-b]indole-3-carboxylic acid was replaced by 1-ethyl-6-methyl-9-(4-fluorobenzyl)-pyrido[3,4-b]indole-3-carboxylic acid.

The third object of the present invention is to provide the use of the aforementioned β-carboline hydrogen sulfide donor derivative in the preparation of drugs for resisting myocardial oxidative damage.

Furthermore, the present invention discloses the use of the aforementioned β-carboline hydrogen sulfide donor derivative in the preparation of ischemic heart disease drugs.

BRIEF DESCRIPTION OF THE DRAWINGS

Figure 1: Synthesis route of compound 3.

DETAILED DESCRIPTION

The specific embodiments of the present invention are further described in detail below, but the present invention is not limited to these embodiments, and any improvement or substitution based on the basic spirit of the present embodiment still falls within the scope of protection required by the claims of the present invention.

Example 1 Preparation of Compound 3 - N-(Benzoylthio)-1-ethyl-9H-pyrido[3,4-b]indole-3-carboxamide

a: Weigh 122 mg of 2.2 mmol of KOH, dissolve in 5 mL of water and place in a 20 mL eggplant-shaped bottle. Stir at room temperature for 5 min, then add 147 mg of 1.3 mmol of hydroxylamine sulfonic acid and 60 mg of 0.44 mmol of thiobenzoic acid. Continue the reaction for 10 min and monitor the reaction by TLC. After 2 h, terminate the reaction, extract with dichloromethane and spin dry to obtain a white solid, which is intermediate 2 (s-acylthiohydroxylamine), with a yield of 67%.

b: Weigh 0.4mmol of intermediate 2 as 100mg and place it in a 25mL eggplant-shaped bottle, dissolve it in 9mL of dry DMF, add 0.8mmol of 1-ethyl-9H-pyridine [3,4-b] indole-3-carboxylic acid 128mg, 0.45mmol of HATU 190mg, 0.8mmol of DIEA 108mg, and react at room temperature. Use TLC to detect the reaction, terminate the reaction after 4h, add 10mL of pure water, extract with ethyl acetate, combine the extracts and backwash with saturated brine, concentrate the organic layer to obtain a crude product, and purify it by normal phase silica gel column chromatography to obtain a white solid, namely compound 3 (N-(benzoylthio)-1-ethyl-9H-pyridine [3,4-b] indole-3-carboxamide), with a yield of 12%.

Example 2

Compound 4: Preparation of N-(benzoylthio)-1-ethyl-9-benzyl-pyridin[3,4-b]indole-3-carboxamide

Referring to the preparation of compound 3, 1-ethyl-9H-pyrido[3,4-b]indole-3-carboxylic acid was replaced with 1-ethyl-9-benzyl-pyrido[3,4-b]indole-3-carboxylic acid, and the yield was 15%.

Example 3

Compound 5 - Preparation of N-(benzoylthio)-1-ethyl-9-(4-fluorobenzyl)-pyrido[3,4-b]indole-3-carboxamide

Referring to the preparation of compound 3, 1-ethyl-9H-pyrido[3,4-b]indole-3-carboxylic acid was replaced with 1-ethyl-9-(4-fluorobenzyl)-pyrido[3,4-b]indole-3-carboxylic acid, and the yield was 36%.

Example 4

Compound 6: Preparation of N-(benzoylthio)-1-ethyl-9-(4-chlorobenzyl)-pyridin[3,4-b]indole-3-carboxamide

Referring to the preparation of compound 3, 1-ethyl-9H-pyrido[3,4-b]indole-3-carboxylic acid was replaced with 1-ethyl-9-(4-chlorobenzyl)-pyrido[3,4-b]indole-3-carboxyl, and the yield was 19%.

Example 5

Compound 7: Preparation of N-(benzoylthio)-1-ethyl-9-(4-methoxybenzyl)-pyridin[3,4-b]indole-3-carboxamide

Referring to the preparation of compound 3, 1-ethyl-9H-pyrido[3,4-b]indole-3-carboxylic acid was replaced with 1-ethyl-9-(4-methoxybenzyl)-pyrido[3,4-b]indole-3-carboxylic acid, and the yield was 24%.

Example 6

Compound 8 - Preparation of N-(benzoylthio)-1-methyl-9-benzyl-pyridinyl[3,4-b]indole-3-carboxamide

Referring to the preparation of compound 3, 1-ethyl-9H-pyrido[3,4-b]indole-3-carboxylic acid was replaced with 1-methyl-9-benzyl-pyrido[3,4-b]indole-3-carboxylic acid, and the yield was 21%.

Example 7

Compound 9: Preparation of N-(benzoylthio)-1-methyl-9-(4-fluorobenzyl)-pyridin[3,4-b]indole-3-carboxamide

Referring to the preparation of compound 3, 1-ethyl-9H-pyrido[3,4-b]indole-3-carboxylic acid was replaced with 1-methyl-9-(4-fluorobenzyl)-pyrido[3,4-b]indole-3-carboxylic acid, and the yield was 21%.

Example 8

Compound 10: Preparation of N-(benzoylthio)-1-methyl-9-(4-chlorobenzyl)-pyridinyl[3,4-b]indole-3-carboxamide

Referring to the preparation of compound 3, 1-ethyl-9H-pyrido[3,4-b]indole-3-carboxylic acid was replaced with 1-methyl-9-(4-chlorobenzyl)-pyrido[3,4-b]indole-3-carboxylic acid, and the yield was 19%.

Example 9

Compound 11——Preparation of N-(benzoylthio)-1-ethyl-9-methyl-pyrido[3,4-b]indole-3-carboxamide

Referring to the preparation of compound 3, 1-ethyl-9H-pyrido[3,4-b]indole-3-carboxylic acid was replaced with 1-ethyl-9-methyl-pyrido[3,4-b]indole-3-carboxylic acid, 31%.

Example 10

Compound 12: Preparation of N-(benzoylthio)-1-ethyl-9-(4-methylbenzyl)-pyrido[3,4-b]indole-3-carboxamide

Referring to the preparation of compound 3, 1-ethyl-9H-pyrido[3,4-b]indole-3-carboxylic acid was replaced with 1-ethyl-9-(4-methylbenzyl)-pyrido[3,4-b]indole-3-carboxylic acid, 24%.

Embodiment 11

Compound 13: Preparation of N-(benzoylthio)-1-ethyl-9-(2-methylbenzyl)-pyrido[3,4-b]indole-3-carboxamide

Referring to the preparation of compound 3, 1-ethyl-9H-pyrido[3,4-b]indole-3-carboxylic acid was replaced with 1-ethyl-9-(2-methylbenzyl)-pyrido[3,4-b]indole-3-carboxylic acid, 27%.

Example 12

Compound 14: Preparation of N-(benzoylthio)-1-ethyl-9-(2-methoxybenzyl)-pyrido[3,4-b]indole-3-carboxamide

Referring to the preparation of compound 3, 1-ethyl-9H-pyrido[3,4-b]indole-3-carboxylic acid was replaced with 1-ethyl-9-(2-methoxybenzyl)-pyrido[3,4-b]indole-3-carboxylic acid, 42%.

Example 13

Compound 15: Preparation of N-(benzoylthio)-1-ethyl-9-(4-trifluoromethylbenzyl)-pyrido[3,4-b]indole-3-carboxamide

Referring to the preparation of compound 3, 1-ethyl-9H-pyrido[3,4-b]indole-3-carboxylic acid was replaced with 1-ethyl-9-(4-trifluoromethylbenzyl)-pyrido[3,4-b]indole-3-carboxylic acid, 36%.

Embodiment 14

Compound 16: Preparation of N-(benzoylthio)-1-ethyl-9-(2-trifluoromethylbenzyl)-pyrido[3,4-b]indole-3-carboxamide

Referring to the preparation of compound 3, 1-ethyl-9H-pyrido[3,4-b]indole-3-carboxylic acid was replaced with 1-ethyl-9-(2-trifluoromethylbenzyl)-pyrido[3,4-b]indole-3-carboxylic acid, 11%.

Embodiment 15

Compound 17: Preparation of N-(benzoylthio)-1-ethyl-9-(2-chlorobenzyl)-pyrido[3,4-b]indole-3-carboxamide

Referring to the preparation of compound 3, 1-ethyl-9H-pyrido[3,4-b]indole-3-carboxylic acid was replaced with 1-ethyl-9-(2-chlorobenzyl)-pyrido[3,4-b]indole-3-carboxylic acid, 17%.

Example 16

Compound 18: Preparation of N-(benzoylthio)-1-ethyl-9-(4-fluorobenzyl)-pyrido[3,4-b]indole-3-carboxamide

Referring to the preparation of compound 3, 1-ethyl-9H-pyrido[3,4-b]indole-3-carboxylic acid was replaced with 1-ethyl-9-(4-fluorobenzyl)-pyrido[3,4-b]indole-3-carboxylic acid, 15%.

Embodiment 17

Compound 19——Preparation of 1-ethyl-6-hydroxy-9-(4-fluorobenzyl)-N-(2-methoxybenzoylthio)-pyrido[3,4-b]indole-3-carboxamide

Referring to the preparation of compound 3, thiobenzoic acid was replaced by o-methoxythiobenzoic acid, and 1-ethyl-9H-pyrido[3,4-b]indole-3-carboxylic acid was replaced by 1-ethyl-6-hydroxy-9-(4-fluorobenzyl)-pyrido[3,4-b]indole-3-carboxylic acid, 22%.

Embodiment 18

Compound 20: Preparation of 1-ethyl-6-methyl-9-(4-fluorobenzyl)-N-(2-fluorobenzoylthio)-pyrido[3,4-b]indole-3-carboxamide

Referring to the preparation of compound 3, thiobenzoic acid was replaced by o-fluorothiobenzoic acid, and 1-ethyl-9H-pyrido[3,4-b]indole-3-carboxylic acid was replaced by 1-ethyl-6-methyl-9-(4-fluorobenzyl)-pyrido[3,4-b]indole-3-carboxylic acid, 15%.

The structural formulas of the compounds of Examples 1-18 are shown in Table 1:

Table 1

Test Example 1

1. MTT assay to detect the toxicity of thiol-triggered hydrogen sulfide donor compounds based on β-carborine structure. The compounds were subjected to the following test:

The toxicity of the example compounds to H9c2 cells was studied by MTT method. Cells (10000 cells/well) were seeded in 96-well plates, and the test compounds (2.5, 5, 10, 20, 40 and 80 μmol/L) were added at 37°C. After incubation for 24 h, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT, Sigma) was added to make the final concentration of each well 0.5 mg/mL. After further incubation for 4 h, the culture medium was removed and 150 μL DMSO was added to each well. The absorbance was measured at 570 nm using a microplate reader, and all measurements were performed at least 3 times under the same conditions. The half inhibitory concentration value (IC ₅₀ ) was calculated using GraphPadPrism 8.

The experimental results are shown in Table 2;

Table 2. Effects of β-carboryl-based thiol-triggered hydrogen sulfide donor compounds (μmol/L) on the survival rate of cardiomyocytes (%) of H9c2 components

In the present application, the thiol-triggered hydrogen sulfide donor compounds 3-20 based on the β-carborine structure, except for compounds 3 and 13 which showed certain toxicity at concentrations higher than 10 μM, the cell survival rates of the remaining compounds were almost close to or greater than 100%, indicating good safety.

2. _{H2O2 -} induced oxidative stress test in cardiomyocytes

H9c2 cells were incubated in a 96-well plate, and the medium was removed after 24 hours. 100 μL of complete medium containing 0, 5, 10, 20, 40, 80 μmol/L of the test compound and 50 μL of H ₂ O ₂ (1.5 mM) were added to each well, with a final concentration of 0.3 mM. After the cells were incubated for another 30 minutes, the survival rate of H9c2 cells was detected by the MTT method.

The test results are shown in Table 3;

Table 3. Effects of pretreatment with β-carborine-based thiol-triggered hydrogen sulfide donor compounds (μmol/L) on the survival rate of H9c2 cardiomyocytes injured by H/R

From the experimental results, it can be concluded that the thiol-triggered hydrogen sulfide donor compounds 5, 9, 10, 11, 13 to 20 based on the β-carborine structure all have significant protective effects on _{H2O2 -} induced oxidative damage, among which compounds 5, 18, 19 and 20 are the most active, which can significantly increase cell activity from about 60% to more than 90%, providing new ideas for the development of drugs for ischemic heart disease.

Claims (6)

1. A β-carboline hydrogen sulfide donor derivative, characterized in that the structural formula of the β-carboline hydrogen sulfide donor derivative is as follows: Among them, _R1 is a proton, an alkyl group, a phenyl group, or an aralkyl group; _R2 is a proton, an alkyl group, a benzyl group, or a substituted benzyl group; _R3 is a proton, an alkyl group, a methoxy group, a hydroxyl group, a nitro group, an amino group, or a halogen group; and _R4 is a proton, an alkyl group, a methoxy group, a hydroxyl group, a nitro group, an amino group, or a halogen group. 2. A β-carboline hydrogen sulfide donor derivative according to claim 1, characterized in that R ₁ is a proton, a methyl group, or an ethyl group; R ₂ is a proton, a methyl group, a benzyl group, an o-fluorobenzyl group, a p-fluorobenzyl group, an o-chlorobenzyl group, a p-chlorobenzyl group, a p-trifluoromethylbenzyl group, an o-trifluoromethylbenzyl group, an o-methylbenzyl group, a p-methylbenzyl group, an o-methoxybenzyl group, or a p-methoxybenzyl group; R ₃ is a proton, a methyl group, a methoxy group, or a hydroxyl group; R ₄ is a proton, a methyl group, a methoxy group, or a hydroxyl group. 3. A β-carboline hydrogen sulfide donor derivative according to claim 1, characterized in that the β-carboline hydrogen sulfide donor derivative is N-(benzoylthio)-1-ethyl-9H-pyrido[3,4-b]indole-3-carboxamide, N-(benzoylthio)-1-ethyl-9-benzyl-pyrido[3,4-b]indole-3-carboxamide, N-(benzoylthio)-1-ethyl-9-(4-fluorobenzyl)-pyrido[3,4-b]indole- 3-carboxamide, N-(benzoylthio)-1-ethyl-9-(4-chlorobenzyl)-pyridin[3,4-b]indole-3-carboxamide, N-(benzoylthio)-1-ethyl-9-(4-methoxybenzyl)-pyridin[3,4-b]indole-3-carboxamide, N-(benzoylthio)-1-methyl-9-benzyl-pyridin[3,4-b]indole-3-carboxamide, N-(benzoylthio)-1-methyl-9-(4-fluorobenzyl)-pyridin[3,4 -b] indole-3-carboxamide, N-(benzoylthio)-1-methyl-9-(4-chlorobenzyl)-pyrido[3,4-b] indole-3-carboxamide, N-(benzoylthio)-1-ethyl-9-methyl-pyrido[3,4-b] indole-3-carboxamide, N-(benzoylthio)-1-ethyl-9-(4-methylbenzyl)-pyrido[3,4-b] indole-3-carboxamide, N-(benzoylthio)-1-ethyl-9-(2-methyl indole-3-carboxamide, N-(benzoylthio)-1-ethyl-9-(2-methoxybenzyl)-pyrido[3,4-b]indole-3-carboxamide, N-(benzoylthio)-1-ethyl-9-(4-trifluoromethylbenzyl)-pyrido[3,4-b]indole-3-carboxamide, N-(benzoylthio)-1-ethyl-9-(2-trifluoromethylbenzyl)-pyrido[3,4-b]indole-3-carboxamide, Any one of N-(benzoylthio)-1-ethyl-9-(2-chlorobenzyl)-pyrido[3,4-b]indole-3-carboxamide, N-(benzoylthio)-1-ethyl-9-(4-fluorobenzyl)-pyrido[3,4-b]indole-3-carboxamide, 1-ethyl-6-hydroxy-9-(4-fluorobenzyl)-N-((2-methoxybenzoyl)thio)-pyrido[3,4-b]indole-3-carboxamide, and 1-ethyl-6-methyl-9-(4-fluorobenzyl)-N-((2-fluorobenzoyl)thio)-pyrido[3,4-b]indole-3-carboxamide. 4. A method for preparing a β-carboline hydrogen sulfide donor derivative according to any one of claims 1 to 3, characterized in that it comprises the following steps: A. Weigh KOH and dissolve it in water, stir at room temperature, add hydroxylamine sulfonic acid and compound 1, stir to react, extract with dichloromethane, and spin dry to obtain intermediate 2; B. Dissolve intermediate 2 in dry DMF, add compound 2, HATU and DIEA, stir the reaction at room temperature, add pure water, extract with ethyl acetate, combine the extracts and backwash with saturated brine, spin dry the organic layer to obtain a crude product, and purify it by normal phase silica gel column chromatography to obtain the product; The compound 1 is a substituted thiobenzoic acid, and its structural formula is: Wherein, R ₄ is a proton, an alkyl group, a methoxy group, a hydroxyl group, a nitro group, an amino group, or a halogen group; The compound 2 is 1-ethyl-9H-pyridine [3,4-b] indole-3-carboxylic acid, 1-ethyl-9-benzyl-pyridine [3,4-b] indole-3-carboxylic acid, 1-ethyl-9-(4-fluorobenzyl)-pyrido [3,4-b] indole-3-carboxylic acid, 1-ethyl-9-(4-chlorobenzyl)-pyridine [3,4-b] indole-3-carboxylic acid, 1-ethyl-9-(4-methoxybenzyl)-pyridine [3,4 -b] indole-3-carboxylic acid, 1-methyl-9-benzyl-pyrido[3,4-b] indole-3-carboxylic acid, 1-methyl-9-(4-fluorobenzyl)-pyrido[3,4-b] indole-3-carboxylic acid, 1-methyl-9-(4-chlorobenzyl)-pyrido[3,4-b] indole-3-carboxylic acid, 1-ethyl-9-methyl-pyrido[3,4-b] indole-3-carboxylic acid, 1-ethyl-9-(4-methylbenzyl)-pyrido[3,4-b] indole-3-carboxylic acid [3,4-b]indole-3-carboxylic acid, 1-ethyl-9-(2-methylbenzyl)-pyrido[3,4-b]indole-3-carboxylic acid, 1-ethyl-9-(2-methoxybenzyl)-pyrido[3,4-b]indole-3-carboxylic acid, 1-ethyl-9-(4-trifluoromethylbenzyl)-pyrido[3,4-b]indole-3-carboxylic acid, 1-ethyl-9-(2-trifluoromethylbenzyl)-pyrido[3,4-b]indole- 3-carboxylic acid, 1-ethyl-9-(2-chlorobenzyl)-pyrido[3,4-b]indole-3-carboxylic acid, 1-ethyl-9-(4-fluorobenzyl)-pyrido[3,4-b]indole-3-carboxylic acid, 1-ethyl-6-hydroxy-9-(4-fluorobenzyl)-pyrido[3,4-b]indole-3-carboxylic acid, 1-ethyl-6-methyl-9-(4-fluorobenzyl)-pyrido[3,4-b]indole-3-carboxylic acid. 5. Use of a β-carboline hydrogen sulfide donor derivative according to any one of claims 1 to 3 in the preparation of a drug for preventing oxidative damage to myocardial cells. 6. Use of a β-carboline hydrogen sulfide donor derivative according to any one of claims 1 to 3 in the preparation of a drug for ischemic heart disease.