阅读说明：本技术 2，3，4-三取代四氢吡咯烷类化合物及其制备方法和应用 (2,3, 4-trisubstituted pyrrolidine compound and preparation method and application thereof ) 是由 刘波 姚庆强 杨皓然 矢倉隆之 李莹 喻琨 陈海蛟 于 2020-06-12 设计创作，主要内容包括：本发明公开了2,3,4-三取代四氢吡咯烷类化合物化合物及其制备方法和应用,具体公开了一类2,3,4-三取代四氢吡咯烷类化合物(I),药理试验证明,本发明的化合物对肺腺癌(A549)、人黑色素瘤细胞(A375)、人结肠癌(LOVO)、人肝癌(HepG2)具有较强的抑制作用,化合物P6对人食管癌(TE-1)具有较强的抑制作用。本发明的化合物对人类风湿性关节炎滑膜细胞具有较强的抑制作用。式(I)<Image he="225" wi="265" file="DDA0002536172130000011.GIF" imgContent="drawing" imgFormat="GIF" orientation="portrait" inline="no"></Image>(The invention discloses 2,3, 4-trisubstituted pyrrolidine compounds and a preparation method and application thereof, and particularly discloses a 2,3, 4-trisubstituted pyrrolidine compound (I). pharmacological tests prove that the compound has a strong inhibitory effect on lung adenocarcinoma (A549), human melanoma cells (A375), human colon cancer (LOVO) and human liver cancer (HepG2), and the compound P6 has a strong inhibitory effect on human esophageal cancer (TE-1). The compound has stronger inhibiting effect on human rheumatoid arthritis synovial cells. Formula (I))

1. A compound shown in a general formula (I) or a pharmaceutically acceptable salt thereof,

r is:

2. the compound or pharmaceutically acceptable salt thereof according to claim 1, wherein R is

3. The compound or pharmaceutically acceptable salt thereof according to claim 2, wherein R is

4. A compound or pharmaceutically acceptable salt thereof according to claim 3, wherein R is

5. A compound or pharmaceutically acceptable salt thereof according to claim 3, wherein R is

6. A process for the preparation of a compound according to any one of claims 1 to 5, or a pharmaceutically acceptable salt thereof, comprising the steps of:

dissolving the compound II in dichloromethane, adding olefin, Grubbs under the protection of nitrogen^2ndHeating and refluxing for 7h, and detecting the disappearance of reaction raw materials by TLC; cooling the reaction liquid to room temperature, filtering the reaction liquid through a short silica gel column, and eluting the reaction liquid with EtOAc to obtain a crude compound (III); dissolving the crude compound (III) in acetonitrile, adding cesium carbonate and thiophenol under the protection of nitrogen, reacting for 3h at 50 ℃, and detecting the disappearance of reaction raw materials by TLC; cooling the reaction liquid to room temperature, filtering the reaction liquid through a short silica gel column, and eluting the reaction liquid with dichloromethane to obtain a crude compound (IV); dissolving the crude compound (IV) in methanol, adding 20% palladium carbon hydroxide and 10% palladium carbon, reacting for 24h under hydrogen condition, filtering the reaction solution with diatomite, and spin-drying the solvent to obtain crude compound (V); and (3) dissolving the crude compound (V) in acetonitrile/water, adding ammonium ceric nitrate, reacting at room temperature for 3 hours, and directly carrying out chromatographic separation on the reaction liquid by using a silica gel column to obtain the target compound.

7. A pharmaceutical composition comprising a compound according to any one of claims 1 to 6, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier or excipient.

8. Use of a compound according to any one of claims 1 to 5 or a pharmaceutically acceptable salt thereof or a pharmaceutical composition according to claim 7 in the manufacture of a medicament for the treatment of cancer.

9. Use according to claim 8, wherein said cancer comprises colon cancer, lung cancer, breast cancer, liver cancer, stomach cancer, lung adenocarcinoma, melanoma, oesophageal cancer.

10. Use of a compound according to any one of claims 1 to 5 or a pharmaceutically acceptable salt thereof or a pharmaceutical composition according to claim 7 in the manufacture of a medicament for the treatment of inflammatory diseases including inflammatory bowel disease, hepatitis, asthma, chronic obstructive pulmonary disease, rheumatoid arthritis or multiple sclerosis.

Technical Field

The invention relates to the field of medicinal chemistry, in particular to a 2,3, 4-trisubstituted pyrrolidine compound, a preparation method thereof, a medicinal preparation containing the compound and a medicinal application of the compound.

Background

The malignant tumor (cancer) has become one of the main public health problems seriously threatening the health of Chinese population, according to the latest statistical data, the death of the malignant tumor accounts for 23.91 percent of the total death causes of residents, and the morbidity and the mortality of the malignant tumor are in a continuously rising state in recent ten years, the medical cost caused by the malignant tumor exceeds 2200 hundred million every year, and the prevention and control situation is severe.

In 2019, month 1, the national cancer center released the latest national cancer statistics of the first phase showing: in 2015, about 392.9 million people with malignant tumor and 233.8 million people with death. On average, over 1 million people per day were diagnosed with cancer, and 7.5 people per minute were diagnosed with cancer. The cancer burden is in a continuously rising state compared to historical data. Over the last 10 years, the incidence of malignancy has remained on the order of 3.9% and mortality has remained 2.5% per year. Lung cancer, liver cancer, upper digestive system tumor, colorectal cancer, female breast cancer and the like are still main malignant tumors in China. Lung cancer is the first disease in men and breast cancer is the first disease in women.

With the increasing progress of urbanization and aging and the change of life style, inflammatory diseases become one of the important threats affecting the global population health, including a plurality of infectious diseases and autoimmune diseases, malignant tumors, cardiovascular diseases, diabetes and other chronic non-infectious major diseases belong to the category of inflammatory diseases, the diseases are spreading in the global scope and become important public health problems seriously harming human health and sustainable development of social economy, and the research is carried out by grasping the common mechanism of a plurality of diseases of inflammation occurrence and development, so that a new strategy is provided for the prevention, diagnosis and treatment of diseases, and the new strategy is more and more paid high attention by various governments and society.

Since the incidence of cancer and inflammatory diseases tends to increase year by year, and the pathological mechanism is very complicated, the discovery of structurally diverse drugs for cancer alleviation is imminent.

Disclosure of Invention

The invention aims to provide a 2,3, 4-trisubstituted pyrrolidine compound, a pharmaceutically acceptable salt thereof, a preparation method thereof, a pharmaceutical composition and application thereof in preparation of medicines for preventing and/or treating diseases related to Sphk1 dysfunction.

In order to solve the technical problem, the invention provides the following technical scheme:

the first aspect of the technical scheme of the invention provides a compound shown as a general formula (I) or a pharmaceutically acceptable salt thereof:

wherein R is

Preferably, the above compound or a pharmaceutically acceptable salt thereof, wherein R is

More preferably, the above compound or a pharmaceutically acceptable salt thereof, wherein R is

Further preferably, the above compound or a pharmaceutically acceptable salt thereof, wherein R is

Still further preferred is the above compound or a pharmaceutically acceptable salt thereof, wherein R is

The pharmaceutically acceptable salt of any one of the compounds is an organic acid salt or an inorganic acid salt, wherein the organic acid comprises acetic acid, trifluoroacetic acid, methanesulfonic acid, toluenesulfonic acid, maleic acid, succinic acid, tartaric acid, citric acid and fumaric acid; the inorganic acid includes hydrochloric acid, hydrobromic acid, nitric acid, sulfuric acid, and phosphoric acid.

In a second aspect of the present invention, there is provided a process for the preparation of a compound of the first aspect, wherein the compound of the general formula (I) of the present invention is prepared by:

R’＝

r is

Dissolving the compound II in dichloromethane, adding the corresponding olefin, Grubbs under the protection of nitrogen^2ndHeating and refluxing for 7h, and detecting the disappearance of reaction raw materials by TLC; cooling the reaction liquid to room temperature, filtering the reaction liquid through a short silica gel column, and eluting the reaction liquid with EtOAc to obtain a crude compound (III); dissolving the crude compound (III) in acetonitrile, adding cesium carbonate and thiophenol under the protection of nitrogen, reacting for 3h at 50 ℃, and detecting the disappearance of reaction raw materials by TLC; cooling the reaction liquid to room temperature, filtering the reaction liquid through a short silica gel column, and eluting the reaction liquid with dichloromethane to obtain a crude compound (IV); dissolving the crude compound (IV) in methanol, adding 20% palladium carbon hydroxide (30 wt%) and 10% palladium carbon (10 wt%), reacting for 24h under hydrogen condition, filtering the reaction solution with diatomite, and spin-drying the solvent to obtain crude compound (V); dissolving the crude compound (V) in acetonitrile/water (volume ratio of 1:4), adding ceric ammonium nitrate, reacting at room temperature for 3h, and directly separating the reaction solution by silica gel column chromatography (eluent is methanol-ethanol-dichloromethane-ammonia water-6: 12:77:5) to obtain the target compound.

The third aspect of the technical scheme of the present invention is to provide a pharmaceutical composition comprising the compound of the first aspect, a pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable carriers and/or diluents, wherein the pharmaceutical composition is any clinically or pharmaceutically acceptable dosage form, preferably an oral preparation or an injection. The physiologically effective amount of the compound represented by the general formula (I) is 0.01g to 10g, and may be 0.01g, 0.015g, 0.02g, 0.025g, 0.03g, 0.04g, 0.05g, 0.1g, 0.125g, 0.2g, 0.25g, 0.3g, 0.4g, 0.5g, 0.6g, 0.75g, 1g, 1.25g, 1.5g, 1.75g, 2g, 2.5g, 3g, 4g, 5g, 6g, 7g, 8g, 9g, 10g, or the like.

Any of the compounds of the present invention, or a pharmaceutically acceptable salt thereof, may be administered orally or parenterally, etc., to a patient in need of such treatment.

For parenteral administration, it can be made into injection. The injection can be prepared by conventional method in pharmaceutical field, and can be aqueous solvent or non-aqueous solvent. The most commonly used aqueous solvent is water for injection, and 0.9% sodium chloride solution or other suitable aqueous solution can also be used; the common non-aqueous solvent is vegetable oil, which is mainly soybean oil for injection, and other aqueous solutions of ethanol, propylene glycol, polyethylene glycol and the like. When preparing the injection, the additive can be not added, and the proper additives can be added according to the property of the medicine, such as osmotic pressure regulator, pH value regulator, solubilizer, filler, antioxidant, bacteriostatic agent, emulsifier, suspending agent and the like. For oral administration, it can be made into conventional solid preparations such as tablet, capsule, pill, granule, etc.; it can also be made into oral liquid, such as oral solution, oral suspension, syrup, etc. When the composition is formulated into oral preparations, appropriate filler, binder, disintegrating agent, lubricant, etc. can be added.

The compound can be added with pharmaceutically acceptable carriers to prepare common medicinal preparations, such as tablets, capsules, powder, syrup, liquid, suspending agents and injection, and common medicinal auxiliary materials such as spices, sweeteners, liquid or solid fillers or diluents and the like can be added.

The clinical administration mode of the compound of the invention can adopt oral administration, injection and other modes.

The compound of the present invention is used in the clinical dosage of 0.01-1000 mg/day, and the dosage may be varied according to the severity of the disease or the dosage form.

The fourth aspect of the present invention provides a use of a compound according to the first aspect and a pharmaceutical composition according to the third aspect for the manufacture of a medicament for the treatment of cancer. The cancer comprises colon cancer, lung cancer, breast cancer, liver cancer, gastric cancer, lung adenocarcinoma, melanoma and esophageal cancer.

In a fourth aspect the present invention provides a use of a compound according to the first aspect and a pharmaceutical composition according to the third aspect in the manufacture of a medicament for the treatment of an inflammatory disease, including inflammatory bowel disease, hepatitis, asthma, chronic obstructive pulmonary disease, rheumatoid arthritis or multiple sclerosis.

The beneficial technical effects are as follows:

pharmacological experiment results show that the compound has a strong inhibition effect on lung adenocarcinoma (A549), human melanoma cells (A375), human colon cancer (LOVO) and human liver cancer (HepG2), and the compound P6 has a strong inhibition effect on human esophageal cancer (TE-1). The compound has stronger inhibiting effect on human rheumatoid arthritis synovial cells.

In addition, the preparation method provided by the invention has the characteristics of no need of intermediate separation, mild reaction conditions, simple operation and the like.

Detailed Description

The present invention will be further described with reference to specific embodiments so that those skilled in the art may better understand the present invention, but the present invention is not limited thereto.

Compound II (0.186mmol) was dissolved in dichloromethane (6mL) and under nitrogen, olefin (0.744mmol), Grubbs, was added^2nd(0.037mmol) and the reaction was heated to reflux for 7h and the disappearance of starting material was detected by TLC. The reaction was cooled to room temperature and filtered through a short silica gel column eluting with EtOAc to give crude compound (III). The crude product (III) was dissolved in acetonitrile (6mL), cesium carbonate (0.372mmol), thiophenol (0.28mmol) were added under nitrogen, and the reaction was carried out at 50 ℃ for 3h, followed by TLC for disappearance of starting material. The reaction was cooled to room temperature and filtered through a short silica gel column eluting with dichloromethane to give crude compound (IV). Dissolving the obtained crude product (IV) in methanol (6mL), adding 20% palladium carbon hydroxide (30 wt%) and 10% palladium carbon (10 wt%), reacting for 24h under hydrogen condition, filtering the reaction solution with diatomite, and spin-drying the solvent to obtain the crude compound (V). The crude product (V) was dissolved in acetonitrile/water (1:4), and cerium ammonium nitrate (0.372mmol) was added to the solution to react at room temperature for 3 hours. Direct use of reaction solutionSeparating with silica gel column chromatography (eluent is methanol-ethanol-dichloromethane-ammonia water-6: 12:77:5) to obtain P series pyrrolidine target compounds. Compound II is structurally illustrated below: