A pyrrolsulfonyl derivative, and its preparation method and application

Abstract

The present invention relates to a new type of pyrrole sulfonyl derivatives, a preparation method thereof, a pharmaceutical composition containing the derivatives, and the use thereof as a therapeutic agent, especially as a gastric acid secretion inhibitor and a potassium ion competitive acid blocker (P-CABs). Specifically, the general structural formula of the pyrrole sulfonyl derivatives is shown in formula (I).

Description

Technical Field

[0001] The present invention relates to a new type of pyrrole sulfonyl derivatives, a preparation method thereof, a pharmaceutical composition containing the derivatives, and the use of the derivatives as therapeutic agents, especially as gastric acid secretion inhibitors and potassium ion competitive acid blockers (P-CABs). Background Art

[0002] In 1987, the first proton pump inhibitor (PPI) omeprazole was successfully developed by AstraZenaca and first launched in Sweden. It is the world's first proton pump inhibitor used in clinical practice and has the strongest effect in inhibiting gastric acid. It is used to treat duodenal ulcers, Zollinger-Ellison syndrome, gastric ulcers and reflux esophagitis. Subsequently, several PPIs products were launched around the world. After years of clinical application, PPIs have become the first choice for the treatment of gastric acid-related diseases. Proton Pump is also called gastric acid pump. Its essence is H + / K + -ATPase (H + / K + -ATPase), which is the gastric secretion of H + The final common pathway is present on the cell membrane of the secretory tubules of the gastric parietal cells, which uses ATP degradation to provide energy for H and K + The first generation of PPIs has a significant inhibitory effect on basal, nocturnal gastric acid, gastric acid secretion stimulated by pentagastrin, and test meals. However, due to limitations in pharmacokinetics and pharmacodynamics, including bioavailability, the effect of administration time on drug efficacy, slow nocturnal acid breakthrough (NAB), instability under acidic conditions (often needing to be prepared into enteral preparations, which take several hours to show effects), and dependence on CYP450 enzymes (significant individual differences in the inhibitory effect of gastric acid secretion), the therapeutic effect and clinical application are affected. Compared with the first generation of PPIs, the new generation of PPIs has obvious advantages in the treatment of gastroesophageal reflux disease (GERD) and other acid-related diseases.

[0003] Potassium-Competitive Acid Blockers (P-CABs) are a new type of acid inhibitors that competitively and reversibly bind to H + Inhibit H + / K +-ATPase activity, and its mechanism of action is significantly different from that of traditional PPIs, so it can be called an acid pump blocker. P-CABs are lipophilic, weakly alkaline, have a high dissociation constant, a long half-life, are stable in acidic conditions, are not primarily metabolized by CYP2C19, and have good tolerance and dependence. In an acidic environment, P-CABs immediately ionize, and the ionized form inhibits H + / K + -ATPase, inhibits H + Transport and acid secretion into the gastric cavity, without the need for microcapsules and microtubules concentrated in the gastric parietal cells and acid activation, can quickly increase the pH value in the stomach, and the enzyme activity is restored after dissociation. After oral administration, humans and animals can absorb it rapidly and reach the peak plasma concentration. Clinical and animal experiments have also shown that P-CABs are faster than PPIs or histamine receptor 2 (Histamine receptor 2, H2) blockers, have a stronger effect on increasing pH, and the blood drug concentration is linearly related to the oral dosage, indicating that this type of drug can achieve the optimal acid suppression state relatively easily and has obvious advantages. Takeda's Vonoprazan Fumarate was approved in Japan in December 2014; it was approved for import in China in December 2019, and some P-CABs preparations have entered clinical research.

[0004] A series of patent applications for P-CABs have been disclosed, including W02005041961, W02006134460, W02009041447 or W02010021149, etc.

[0005] Although a series of P-CABs drugs or compounds have been disclosed, there is still a need to develop new compounds with richer structural types, better efficacy and greater safety. After continuous efforts, the present invention designs compounds having a structure shown in general formula (I), and finds that compounds having such a structure exhibit excellent effects and good safety. Summary of the invention

[0006] In order to solve the above problems in the prior art, the object of the present invention is to provide a new pyrrole sulfonyl derivative, its tautomer or its stereoisomer, and a pharmaceutically acceptable salt thereof, so as to screen out compounds having excellent performance in terms of effectiveness, safety and selectivity for use as gastric acid secretion inhibitors and potassium ion competitive acid blockers (P-CABs).

[0007] Another object of the present invention is to provide a method for preparing the derivative, its tautomer or stereoisomer, and its pharmaceutically acceptable salt.

[0008] In order to achieve the purpose of the invention, the present invention adopts the following technical solutions:

[0009] In a first aspect, the present invention provides a pyrrole sulfonyl derivative, its tautomer or stereoisomer, and a pharmaceutically acceptable salt thereof. The structure of the pyrrole sulfonyl derivative is shown in formula (I):

[0010]

[0011] in:

[0012] A is phenyl or pyridyl;

[0013] Dashed lines represent optional bonds;

[0014] R 1 Selected from H, C 1-6 Alkyl, C 3-12 cycloalkyl, 3-12 membered heterocyclyl, 5-12 membered aryl or 5-12 membered heteroaryl, C 1-6 Alkyl-NH-C 1-6 Alkyl, C 1-6 Alkyl-OC 1-6 Alkyl, -C(=O)NHR a 、-C(=O)R b 、-S(=O) m -C 1-6 alkyl;

[0015] Wherein, the alkyl, cycloalkyl, heterocyclic, aryl or heteroaryl group may be further optionally substituted with 1-3 hydroxyl, halogen, C 1-6 Alkyl, C 1-6 Alkoxy substituted;

[0016] R 2 Selected from H, C 1-6 Alkyl, C 1-6 Alkoxy, halogen, cyano, C 2-6 Alkynyl, C 2-6 Alkenyl, -OC 1-6 Alkyl-OC 1-6 Alkyl, -NR c R d ;

[0017] R 3 is selected from halogen, 5-8 membered aryl, 5-8 membered heteroaryl, cyano, C 2-6 Alkynyl, C 2-6 Alkenyl, -OC 1-6 Alkyl-OC 1-6 Alkyl, -NR c R d ;

[0018] Or two R 3 Together with the carbon atoms to which they are attached, they form a fused ring aromatic group, wherein the aromatic group, heteroaromatic group and fused ring aromatic group may be further optionally substituted by 1-3 C 1-6 Alkyl, C 1-6 Alkoxy, halogen, C 2-6 Alkynyl, C 2-6 Alkenyl, cyano, -OC 1-6 Alkyl-OC 1-6 Alkyl, -NR c R d replaced by;

[0019] R 4 Selected from C 1-6 wherein the alkyl group may be optionally further substituted by 1-3 deuterium or halogen;

[0020] n is selected from 0, 1, 2, 3;

[0021] m is selected from 0, 1, 2;

[0022] R a , R b , R c , R d Each independently selected from C 1-6 Alkyl, C 1-6 Alkoxy, C 3-7 of a cycloalkyl group.

[0023] In some embodiments: R 1 Selected from H, C 1-6 Alkyl, C 3-12 Cycloalkyl, C 1-6 Alkyl-NH-C 1-6 Alkyl, C 1-6 Alkyl-OC 1-6 Alkyl, -C(=O)NHR a 、-C(=O)OR b ;

[0024] Wherein, the alkyl, cycloalkyl, heterocyclic group, may be further optionally substituted with halogen, C 1-6 Alkyl substituted.

[0025] In some embodiments: Ring A is selected from phenyl;

[0026] R 1 Selected from C 1-6 Alkyl-NH-C 1-6 Alkyl, C 1-6 Alkyl-OC 1-6 Alkyl, C 3-12 Cycloalkyl or -C(=O)NHRa .

[0027] In some embodiments, the pyrrole sulfonyl derivative has a general structural formula as shown in formula (II):

[0028]

[0029] In some preferred embodiments, the R 2 Selected from H, C 1-6 Alkoxy, halogen.

[0030] In some preferred embodiments, the R 3 is selected from halogen; R 4 Selected from C 1-6 of alkyl.

[0031] In some preferred embodiments, the pyrrole sulfonyl derivative has a general structural formula as shown in formula (III):

[0032]

[0033] The present invention provides a pharmaceutically acceptable salt of the pyrrole sulfonyl derivatives, wherein the salt may be a hydrochloride or a trifluoroacetate.

[0034] Furthermore, the pyrrole sulfonyl derivatives of the present invention can be selected from any one of the following structures:

[0035]

[0036]

[0037] In another aspect, the present invention provides a pharmaceutical composition, comprising the pyrrole sulfonyl derivatives, their tautomers or stereoisomers, and pharmaceutically acceptable salts thereof, and pharmaceutically acceptable carriers and / or excipients as described above.

[0038] In another aspect, the present invention provides a pyrrole sulfonyl derivative, its tautomer or stereoisomer, and a pharmaceutically acceptable salt thereof as described in the first aspect, and a pharmaceutical composition as described in the third aspect for use in the preparation of a gastric acid secretion inhibitor, H + / K + - Use in ATPase inhibitors or potassium ion competitive acid blockers.

[0039] The present invention provides a use of the above-mentioned pyrrole sulfonyl derivative, its tautomer or its stereoisomer, and its pharmaceutically acceptable salt or the above-mentioned pharmaceutical composition in the preparation of drugs for treating or preventing the following diseases: peptic ulcer, Zollinger-Ellison syndrome, gastric erosive esophagitis, reflux esophagitis, symptomatic gastroesophageal reflux disease, Barrett's esophagitis, functional dyspepsia, Helicobacter pylori infection, gastric cancer, gastric MALT lymphoma, ulcers caused by non-steroidal anti-inflammatory drugs, hyperacidity caused by postoperative stress or ulcers caused by postoperative stress; or use in the preparation of drugs for inhibiting upper gastrointestinal bleeding caused by peptic ulcer, acute stress ulcer, hemorrhagic gastritis or invasive stress. The pyrrole sulfonyl derivative provided by the present invention has small toxic and side effects and excellent safety; at the same time, it has good pharmacokinetic properties, a longer half-life, and a more sustained acid suppression effect, and is expected to have a better improvement effect on the nocturnal acid breakthrough phenomenon.

[0040] Explanation of terms

[0041] Unless otherwise stated, some of the terms used in the specification and claims of the present invention are defined as follows:

[0042] "Cycloalkyl" refers to a saturated or partially unsaturated monocyclic or polycyclic cyclic hydrocarbon substituent, the cycloalkyl ring includes 3 to 20 carbon atoms, preferably includes 3 to 12 carbon atoms, more preferably includes 3 to 6 carbon atoms. Non-limiting examples of monocyclic cycloalkyl include, but are not limited to cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, cycloheptyl, cycloheptatrienyl, cyclooctyl, etc.; polycyclic cycloalkyl includes cycloalkyl of spiro ring, condensed ring and bridge ring. Cycloalkyl can be optionally substituted or unsubstituted.

[0043] "Heterocyclyl", "heterocycle" or "heterocyclic" are used interchangeably in this application and refer to a saturated or partially unsaturated monocyclic, bicyclic or tricyclic non-aromatic heterocyclic group containing 3-12 ring atoms, wherein at least one ring atom is a heteroatom, such as an oxygen, nitrogen, sulfur atom, etc. Preferably, it has a 5-7 membered monocyclic ring or a 7-10 membered bi- or tricyclic ring, which may contain 1, 2 or 3 atoms selected from nitrogen, oxygen and / or sulfur. Examples of "heterocyclyl" include, but are not limited to, morpholinyl, oxetanyl, thiomorpholinyl, tetrahydropyranyl, 1,1-dioxo-thiomorpholinyl, piperidinyl, 2-oxo-piperidinyl, pyrrolidinyl, 2-oxo-pyrrolidinyl, piperazin-2-one, 8-oxa-3-aza-bicyclo[3.2.1]octyl and piperazinyl. The heterocyclyl ring may be fused to an aryl, heteroaryl or cycloalkyl ring, wherein the ring attached to the parent structure is the heterocyclyl. The heterocyclyl may be optionally substituted or unsubstituted.

[0044] "Aryl" refers to a carbocyclic aromatic system containing one or two rings, wherein the rings may be fused together. The term "aryl" includes aromatic groups such as phenyl, naphthyl, tetrahydronaphthyl. Preferably, aryl is C 6-10 Aryl, more preferably phenyl and naphthyl, most preferably phenyl. Aryl can be substituted or unsubstituted. The "aryl" can be fused with heteroaryl, heterocyclic or cycloalkyl, wherein the aryl ring is connected to the parent structure, non-limiting examples include but are not limited to:

[0045]

[0046] "Heteroaryl" refers to an aromatic 5 to 6-membered monocyclic or 9 to 10-membered bicyclic ring, which may contain 1 to 4 atoms selected from nitrogen, oxygen and / or sulfur. The embodiment of "heteroaryl" includes, but is not limited to, furyl, pyridyl, 2-oxo-1,2-dihydropyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, thienyl, isoxazolyl, oxazolyl, oxadiazolyl, imidazolyl, pyrrolyl, pyrazolyl, triazolyl, tetrazolyl, thiazolyl, isothiazolyl, 1,2,3-thiadiazolyl, benzodioxolyl, benzimidazolyl, indolyl, isoindolyl, 1,3-dioxo-isoindolyl, quinolyl, indazolyl, benzisothiazolyl, benzoxazolyl and benzisoxazolyl. Heteroaryl may be optionally substituted or unsubstituted. The heteroaryl ring may be fused to an aryl, heterocyclyl or cycloalkyl ring, wherein the ring connected to the parent structure is a heteroaryl ring, non-limiting examples include but are not limited to:

[0047]

[0048] "Optional" means that the event it describes may but need not occur. For example, "A is optionally replaced by 1 to more R 2 The term "substituted" means that the group A can be replaced by 1 to more than one R 2 Replaced or not by R 2 Replacement situation.

[0049] The present invention provides a pyrrole sulfonyl derivative with a new structure. The test results show that the pyrrole sulfonyl derivative exhibits excellent gastric acid secretion inhibition and potassium ion competitive acid blocker (P-CABs) activity, and can be used to prepare drugs for treating or preventing peptic ulcer, Zollinger-Ellison syndrome, gastric erosive esophagitis, reflux esophagitis, symptomatic gastroesophageal reflux disease, Barrett's esophagitis, functional dyspepsia, Helicobacter pylori infection, gastric cancer, gastric MALT lymphoma, ulcers caused by non-steroidal anti-inflammatory drugs, hyperacidity caused by postoperative stress or ulcers caused by postoperative stress; or in the preparation of drugs for inhibiting upper gastrointestinal bleeding caused by septic ulcers, acute stress ulcers, hemorrhagic gastritis or invasive stress. The pyrrole sulfonyl derivative provided by the present invention has small toxic and side effects and excellent safety; at the same time, it has good pharmacokinetic properties, a long half-life, and a more sustained acid suppression effect, and is expected to have a better improvement effect on the nocturnal acid breakthrough phenomenon. BRIEF DESCRIPTION OF THE DRAWINGS

[0050] Figure 1 Instruct voltage program for manual patch clamp hERG current testing. DETAILED DESCRIPTION

[0051] The method of the present invention is described below by means of specific embodiments to make the technical solution of the present invention easier to understand and grasp, but the present invention is not limited thereto. 1 HNMR spectra were measured using a Bruker instrument (400 MHz), and chemical shifts were expressed in ppm using tetramethylsilane as an internal standard (0.00 ppm). 1 HNMR notation: s = singlet, d = doublet, t = triplet, q = quartet, m = multiplet, br = broadened, dd = doublet of a doublet, dt = doublet of a triplet. Coupling constants, when given, are given in Hz.

[0052] The mass spectrum was obtained by LC / MS using ESI as the ionization method.

[0053] In the following examples, unless otherwise indicated, all temperatures are in degrees Celsius. Unless otherwise indicated, various starting materials and reagents are commercially available or synthesized according to known methods, and commercially available materials and reagents are used directly without further purification.

[0054] CD3OD: deuterated methanol;

[0055] CDCl3: deuterated chloroform;

[0056] DMSO-d6: deuterated dimethyl sulfoxide;

[0057] Hydrogen atmosphere means that the reaction bottle is connected to a hydrogen balloon with a volume of about 1L.

[0058] Unless otherwise specified in the examples, the solution in the reaction refers to an aqueous solution.

[0059] Unless otherwise specified in the examples, the reaction temperature is room temperature, 20°C-30°C.

[0060] Preparation of intermediates

[0061] Synthesis of intermediate Int 1

[0062]

[0063] Step 1: Synthesis of 1-(5-(2-fluorophenyl)-1H-pyrrol-3-yl)-N-methylmethanamine (Int 1-2)

[0064] At room temperature, compound Int 1-1 (3.8 g, 20.1 mmol) was dissolved in 30% methylamine alcohol solution (20 mL), stirred for 1 hour, and then NaBH4 (2.3 g, 60.3 mmol) was added in batches, and stirring was continued for 1 hour. Water (20 mL) was added, and the mixture was extracted with ethyl acetate (100 mL). The organic phase was washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain 4.1 g of a yellow solid, with a yield of 97.6%.

[0065] Step 2: Synthesis of tert-butyl ((5-(2-fluorophenyl)-1H-pyrrol-3-yl)methyl)(methyl)carbamate (Int1-3)

[0066] At room temperature, compound Int 1-2 (4.1 g, 20.6 mmol) was dissolved in dichloromethane (50 mL), and triethylamine (11 mL, 78.9 mmol) and Boc2O (9.0 g, 41.2 mmol) were added respectively. The mixture was reacted at room temperature for 2 hours, concentrated under reduced pressure, and the crude product was separated and purified by column chromatography (n-hexane / ethyl acetate) to obtain 5.2 g of a white solid with a yield of 82.5%.

[0067] Step 3: Synthesis of tert-butyl ((1-((3-bromophenyl)sulfonyl)-5-(2-fluorophenyl)-1H-pyrrol-3-yl)methyl)(methyl)carbamate (Int 1)

[0068] At room temperature, compound Int 1-3 (3 g, 9.86 mmol) was dissolved in DMF (50 mL), cooled to 0°C, sodium hydride (600 mg, 14.8 mmol) was added, and after stirring at room temperature for 10 minutes, 3-bromobenzenesulfonyl chloride (3 g, 11.8 mmol) was added. The reaction was continued for 1 hour, water (50 mL) was added to quench the reaction, and the mixture was extracted with ethyl acetate (100 mL). The organic phase was washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was separated and purified by column chromatography (n-hexane / ethyl acetate) to obtain 4.8 g of a white solid with a yield of 93.0%.

[0069] Synthesis of intermediate Int 2

[0070]

[0071] Synthesis of tert-butyl ((1-((5-bromopyridin-3-yl)sulfonyl)-5-(2-fluorophenyl)-1H-pyrrol-3-yl)methyl)(methyl)carbamate (Int 2)

[0072] In an ice bath, compound Int 1-3 (3.0 g, 9.87 mmol) was dissolved in DMF (40 ml), and then NaH (789 mg, 19.71 mmol) was added, stirred for half an hour, and then compound Int 2-1 (3.03 g, 11.83 mmol) was added, and stirring was continued for 2 hours. After the reaction was complete, ethyl acetate (150 mL) was added for dilution, and the mixture was washed twice with saturated brine (100 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. Purification by column chromatography (PE / EA=3:1) gave 4.5 g of a yellow oily compound with a yield of 87.2%.

[0073] Synthesis of intermediate Int 3

[0074]

[0075] Step 1: Synthesis of 1-(5-bromo-1H-pyrrol-3-yl)-N-methylmethanamine (Int 3-2)

[0076] At room temperature, compound Int 3-1 (5.0 g, 28.74 mmol) was dissolved in a methanol solution of methylamine (50 mL), stirred at room temperature for 2 h, then cooled in an ice-water bath, sodium borohydride (2.2 g, 57.47 mmol) was added, and stirring was continued for 2 h. After the reaction was completed, a saturated aqueous solution of ammonium chloride was added to quench the reaction, extracted with dichloromethane (100 mL × 2), the combined organic phases were washed with saturated brine (150 mL × 1), the organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain 5.43 g of a yellow oily compound, which was directly used for the next step without purification, [M+H] + :189.1.

[0077] Step 2: Synthesis of tert-butyl ((5-bromo-1H-pyrrol-3-yl)methyl)(methyl)carbamate (Int 3-3)

[0078] At room temperature, compound Int 3-2 (15.43 g, 28.72 mmol) and triethylamine (7.98 mL, 57.44 mmol) were dissolved in dichloromethane (50 mL), and then Boc2O (7.52 g, 34.47 mmol) was slowly added and stirred at room temperature for 3 h. After the reaction was completed, the reaction solution was concentrated under reduced pressure and purified by column chromatography (PE / EA=5:1) to obtain 8.2 g of a colorless oily compound with a yield of 98.60%, [M+H] + :289.1.

[0079] Step 3: Synthesis of tert-butyl ((5-(2,4-difluorophenyl)-1H-pyrrol-3-yl)methyl)(methyl)carbamate (Int 3-5)

[0080] Compound Int 3-3 (8.0 g, 27.67 mmol), Int 3-4 (8.0 g, 33.20 mmol), Pd(dppf)Cl2 (2.1 g, 2.77 mmol), and potassium carbonate (9.6 g, 69.16 mmol) were dissolved in 1,4-dioxane and water (80 / 16 mL), and stirred at 115 ° C for 4 h under nitrogen protection. After the reaction was completed, it was extracted with dichloromethane (100 mL×2), and the combined organic phases were washed with saturated brine (150 mL×1), and the organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. Purification by column chromatography (PE / EA=3:1) gave 6.2 g of a brown oily compound with a yield of 69.66%, [M+H] + :323.2.

[0081] Step 4: Synthesis of tert-butyl ((1-((3-bromophenyl)sulfonyl)-5-(2,4-difluorophenyl)-1H-pyrrol-3-yl)methyl)(methyl)carbamate (Int 3)

[0082] In an ice bath, compound Int 3-5 (2.0 g, 6.20 mmol) was dissolved in DMF (15 mL), sodium hydride (373 mg, 9.31 mmol) was added, and the mixture was stirred for 30 minutes. Compound Int 3-6 (1.9 g, 7.45 mmol) was then added to the reaction system, and the mixture was stirred for 2 h. After the reaction was completed, a saturated aqueous solution of ammonium chloride was added to quench the reaction, and the mixture was extracted with ethyl acetate (30 mL × 2). The combined organic phases were washed with saturated brine (50 mL × 1), and the organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. 3.0 g of a brown oily compound was obtained by column chromatography (PE / EA = 3:1) with a yield of 89.55%.

[0083] Synthesis of intermediate Int 4

[0084]

[0085] Step 1: Synthesis of 1,3-dibromo-5-(3-methoxypropoxy)benzene (Int 4-3)

[0086] At room temperature, compound Int 4-1 (3.0 g, 11.90 mmol), Int 4-2 (1.55 g, 14.28 mmol) and potassium carbonate (4.11 g, 29.78 mmol) were dissolved in DMF (30 mL) and stirred overnight at 100° C. After the reaction was complete, water (100 mL) was added, and the mixture was extracted with ethyl acetate (50 mL×2). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure and purified by column chromatography (PE / EA=5:1) to obtain 3.5 g of a colorless oily compound with a yield of 92.11%.

[0087] Step 2: Synthesis of benzyl(3-bromo-5-(3-methoxypropoxy)phenyl)sulfane (Int 4-5)

[0088] At room temperature, compound Int 4-3 (3.5 g, 10.80 mmol), Int 4-4 (1.34 g, 10.80 mmol), Pd2(dba)3, (495 mg, 0.54 mmol), xantphos (313 mg, 0.54 mmol) and DIEA (3.5 g, 27.13 mmol) were dissolved in toluene (40 mL) and stirred at 100° C. for 4 h. After the reaction was complete, the mixture was concentrated under reduced pressure and purified by column chromatography (PE / EA=10:1) to obtain 3.0 g of a colorless oily compound with a yield of 75.02%.

[0089] Step 3: Synthesis of 3-bromo-5-(3-methoxypropoxy)benzenesulfonyl chloride (Int 4-7)

[0090] At room temperature, compound Int 4-5 (2.5 g, 6.81 mmol) and Int 4-6 (2.68 g, 13.61 mmol) were dissolved in a solution of acetonitrile (69 mL), water (4.15 mL) and acetic acid (4.15 mL), and the mixture was reacted at room temperature for 1 h. After the reaction was complete, water (100 mL) was added, and the mixture was extracted with ethyl acetate (50 mL×2). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The filtrate was separated and purified by column chromatography (PE / EA=5:1) to obtain 1.1 g of a yellow oil with a yield of 47.03%. [M+H] + :344.9.

[0091] Step 4: Synthesis of tert-butyl ((1-((3-bromo-5-(3-methoxypropoxy)phenyl)sulfonyl)-5-(2-fluorophenyl)-1H-pyrrol-3-yl)methyl)(methyl)carbamate (Int 4)

[0092] At room temperature, sodium hydride (160.4 mg, 4.00 mmol) was added to a DMF (20 mL) solution. Compound Int 1-3 (811.93 mg, 2.67 mmol) was added to the above solution and stirred at room temperature for 0.5 h. Compound Int 4-7 (1.1 g, 3.20 mmol) was slowly added to the reaction solution, and reacted at room temperature for 2 h under nitrogen protection. After the reaction was complete, water (100 mL) was added, and the mixture was extracted with ethyl acetate (30 mL × 2). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure and purified by column chromatography (PE / EA = 3:1) to obtain 600 mg of a yellow viscous solid with a yield of 36.87%, [M+H] + :613.1.

[0093] Example 1

[0094] Synthesis of 1-(5-(2-fluorophenyl)-1-((3-(3-methoxyprop-1-yn-1-yl)phenyl)sulfonyl)-1H-pyrrol-3-yl)-N-methylmethanamine hydrochloride (Ⅱ-1)

[0095]

[0096] Step 1: Synthesis of tert-butyl ((5-(2-fluorophenyl)-1-((3-(3-methoxyprop-1-yn-1-yl)phenyl)sulfonyl)-1H-pyrrol-3-yl)methyl)(methyl)carbamate (1-2)

[0097] Add compound Int 1 (260 mg, 0.5 mmol), compound 1-1 (70 mg, 1 mmol), Pd(PPh3)2Cl2 (35 mg, 0.05 mmol), CuI (19 mg, 0.1 mmol), TEA (150 mg, 1.5 mmol), DMF (3 mL) into a sealed tube, stir, replace with nitrogen 3 times, heat to 80 ° C and stir for 4 h. TLC spot plate monitoring, the reaction is complete, and new spots are generated (PE / EA = 3 / 1, Rf = 0.3). Add 30 mL of water, extract with ethyl acetate (30 mL × 2), combine the organic phases, wash once with saturated brine (30 mL), dry the organic phase with anhydrous sodium sulfate, control the temperature at 45 ° C and concentrate under reduced pressure, separate by column chromatography (PE / EA = 3:1 elution) to obtain 220 mg of colorless oil.

[0098] Step 2: Synthesis of 1-(5-(2-fluorophenyl)-1-((3-(3-methoxypropyl-1-yn-1-yl)phenyl)sulfonyl)-1H-pyrrol-3-yl)-N-methylmethanamine hydrochloride (II-1)

[0099] At room temperature, compound 1-2 (220 mg) and dichloromethane (10 mL) were added to the reaction bottle, stirred to dissolve, 4M hydrochloric acid / 1,4-dioxane (4 mL) was added, and the reaction was carried out at room temperature for 1 h. The reaction was completed and a new spot was generated (DCM / MeOH=10:1, Rf=0.2) after TLC spot plate monitoring. The mixture was dried by rotation, and n-hexane was added. Solid precipitated, which was filtered and dried to obtain 100 mg of off-white solid.

[0100] 1 HNMR: (400MHz, DMSO-d6) δ9.15 (s, 2H), 7.85-7.78 (m, 2H), 7.60 (t, J=8.0Hz, 1H), 7.56-7.47 (m, 2H), 7.36 (s, 1 H), 7.28-7.19 (m, 2H), 7.08 (t, J=1.6Hz, 1H), 6.57 (s, 1H), 4.36 (s, 2H), 3.99 (s, 2H), 3.36 (s, 3H), 3.34 (s, 3H).

[0101] Example 2

[0102] Synthesis of 1-(5-(2-fluorophenyl)-1-((5-(3-methoxyprop-1-yn-1-yl)pyridin-3-yl)sulfonyl)-1H-pyrrol-3-yl)-N-methylamine hydrochloride (Ⅱ-2)

[0103]

[0104] Step 1: Synthesis of tert-butyl ((5-(2-fluorophenyl)-1-((5-(3-methoxyprop-1-yn-1-yl)pyridin-3-yl)sulfonyl)-1H-pyrrol-3-yl)methyl))(methyl)carbamate (2-2)

[0105] Add compound Int 2 (260 mg, 0.5 mmol), 1-1 (70 mg, 1 mmol), Pd(PPh3)2Cl2 (35 mg, 0.05 mmol), CuI (19 mg, 0.1 mmol), TEA (150 mg, 1.5 mmol), DMF (3 mL) into a sealed tube, stir, replace with nitrogen 3 times, heat to 80 ° C and stir for 4 h. TLC spot plate monitoring, the reaction is complete, and new spots are generated (PE / EA = 3 / 1, Rf = 0.3). Add 30 mL of water, extract with ethyl acetate (30 mL × 2), combine the organic phases, wash once with saturated brine (30 mL), dry the organic phase with anhydrous sodium sulfate, control the temperature at 45 ° C and concentrate under reduced pressure, separate by column chromatography (PE / EA = 3:1 elution) to obtain 220 mg of brown oil.

[0106] Step 2: Synthesis of 1-(5-(2-fluorophenyl)-1-((5-(3-methoxyprop-1-yn-1-yl)pyridin-3-yl)sulfonyl)-1H-pyrrol-3-yl)-N-methylamine hydrochloride (II-2)

[0107] At room temperature, compound 2-2 (220 mg) and dichloromethane (10 mL) were added to the reaction bottle, stirred to dissolve, 4M hydrochloric acid / 1,4-dioxane (4 mL) was added, and the reaction was carried out at room temperature for 1 h. The reaction was completed by TLC spot plate monitoring, and a new spot was generated (DCM / MeOH=10 / 1, Rf=0.2). The mixture was spin-dried, ethyl acetate (2 mL) was added to dissolve, and n-hexane (10 mL) was added. Solid precipitated, filtered, and dried to obtain 130 mg of brown solid.

[0108] 1 HNMR: (400MHz, DMSO-d6) δ9.17 (s, 2H), 8.99 (s, 1H), 8.55 (s, 1H), 7.87 (d, J=1.6Hz, 2H), 7.60-7.53 (m, 1H) , 7.32-7.22(m, 2H), 7.13(t, J=7.2, 1H), 6.62(s, 1H), 4.40(s, 2H), 4.00(s, 2H), 3.57(s, 3H), 3.37(s, 3H).

[0109] Example 3

[0110] Synthesis of 4-(5-((2-(2-fluorophenyl)-4-((methylamino)methyl)-1H-pyrrol-1-yl)sulfonyl)pyridin-3-yl)-2-methylbut-3-yn-2-ol (Ⅱ-3)

[0111]

[0112] Step 1: Synthesis of tert-butyl ((5-(2-fluorophenyl)-1-((5-(3-hydroxy-3-methylbut-1-yn-1-yl)pyridin-3-yl)sulfonyl)-1H-pyrrol-3-yl)methyl)(methyl)carbamate (3-2)

[0113] Add compound Int 2 (210 mg, 0.4 mmol), compound 3-1 (68 mg, 0.8 mmol), Pd(PPh3)2Cl2 (28 mg, 0.04 mmol), CuI (16 mg, 0.08 mmol), TEA (122 mg, 1.2 mmol), DMF (3 mL) to the sealed tube, stir, replace with nitrogen 3 times, heat to 80 ° C and stir for 4 h. TLC spot plate monitoring, the reaction is complete, and new spots are generated (PE / EA = 3: 1, Rf = 0.3). Add 30 mL of water, extract with ethyl acetate (30 mL × 2), combine the organic phases, wash once with saturated brine (30 mL), dry the organic phase with anhydrous sodium sulfate, control the temperature at 45 ° C and concentrate under reduced pressure, separate by column chromatography (PE / EA = 3: 1) to obtain 200 mg of colorless oil.

[0114] Step 2: Synthesis of 4-(5-((2-(2-fluorophenyl)-4-((methylamino)methyl)-1H-pyrrol-1-yl)sulfonyl)pyridin-3-yl)-2-methylbut-3-yn-2-ol (II-3)

[0115] At room temperature, compound 3-2 (200 mg) and dichloromethane (4 mL) were added to the reaction bottle, stirred to dissolve, 4M hydrochloric acid / 1,4-dioxane (2 mL) was added, and the reaction was carried out at room temperature for 1 h. The reaction was completed and a new spot was generated (DCM / MeOH=10:1, Rf=0.2) by TLC spot plate monitoring. Sodium bicarbonate aqueous solution was added to adjust the pH to 8, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine twice, dried over anhydrous sodium sulfate, and separated on a TLC plate to obtain 50 mg of a brown solid.

[0116] 1HNMR: (400MHz, CD3OD) δ8.78 (s, 1H), 8.49 (s, 1H), 7.70 (s, 1H), 7.57 (s, 1H), 7.55-7.49 (m, 1H ), 7.28-7.21(m, 1H), 7.18-7.10(m, 2H), 6.39(s, 1H), 3.70(s, 2H), 2.44(s, 3H), 1.60(s, 6H).

[0117] Example 4

[0118] Synthesis of 1-(1-((5-(cyclopropylethynyl)pyridin-3-yl)sulfonyl)-5-(2-fluorophenyl)-1H-pyrrol-3-yl)-N-methylmethanamine (Ⅱ-4)

[0119]

[0120] Step 1: Synthesis of tert-butyl ((1-((5-(cyclopropylethynyl)pyridin-3-yl)sulfonyl)-5-(2-fluorophenyl)-1H-pyrrol-3-yl)methyl)(methyl)carbamate (4-2)

[0121] Add compound Int 2 (210 mg, 0.4 mmol), compound 4-1 (53 mg, 0.8 mmol), Pd(PPh3)2Cl2 (28 mg, 0.04 mmol), CuI (16 mg, 0.08 mmol), TEA (122 mg, 1.2 mmol), DMF (3 mL) into a sealed tube, stir, replace with nitrogen 3 times, heat to 80 ° C and stir for 4 h. TLC spot plate monitoring, the reaction is complete, and new spots are generated (PE / EA = 3: 1, Rf = 0.3). Add 30 mL of water, extract with ethyl acetate (30 mL × 2), combine the organic phases, wash once with saturated brine (30 mL), dry the organic phase with anhydrous sodium sulfate, control the temperature at 45 ° C and concentrate under reduced pressure, separate by column chromatography (PE / EA = 3: 1) to obtain 200 mg of brown oil.

[0122] Step 2: Synthesis of 1-(1-((5-(cyclopropylethynyl)pyridin-3-yl)sulfonyl)-5-(2-fluorophenyl)-1H-pyrrol-3-yl)-N-methylmethanamine (II-4)

[0123] At room temperature, compound 4-2 (200 mg) and dichloromethane (4 mL) were added to the reaction bottle, stirred to dissolve, 4M hydrochloric acid / 1,4-dioxane (2 mL) was added, and the reaction was carried out at room temperature for 1 h. The reaction was completed and a new spot was generated (DCM / MeOH=10:1, Rf=0.2) by TLC spot plate monitoring. Sodium bicarbonate aqueous solution was added to adjust the pH to 8, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine twice, dried over anhydrous sodium sulfate, and separated on a TLC plate to obtain 80 mg of brown oil.

[0124] 1 HNMR: (400MHz, CD3OD) δ8.71 (s, 1H), 8.41 (s, 1H), 7.66 (s, 1H), 7.60 (s, 1H), 7.54-7.48 (m, 1H), 7.22-7.0 9(m, 3H), 6.38(s, 1H), 3.70(s, 2H), 2.44(s, 3H), 1.60-1.50(m, 1H), 1.03-0.95(m, 2H), 0.88-0.81(m, 2H).

[0125] Example 5

[0126] Synthesis of 1-(5-(2-fluorophenyl)-1-((3-(3-methylbut-1-yn-1-yl)phenyl)sulfonyl)-1H-pyrrol-3-yl)-N-methylmethanamine (Ⅱ-5)

[0127]

[0128] Step 1: Synthesis of tert-butyl ((5-(2-fluorophenyl)-1-((3-(3-methylbut-1-yn-1-yl)phenyl)sulfonyl)-1H-pyrrol-3-yl)methyl)(methyl)carbamate (5-1)

[0129] Under nitrogen protection at room temperature, compound Int 1 (500 mg, 0.960 mmol) was dissolved in DMF (5 mL), and Pd(PPh3)2Cl2 (67 mg, 0.096 mmol), CuI (19 mg, 0.096 mmol), triethylamine (0.5 mL, 3.84 mmol) and 3-methylbut-1-yne (260 mg, 3.84 mmol) were added respectively, and the mixture was sealed, heated to 80°C, and stirred overnight. After the reaction was complete, the reaction solution was concentrated under reduced pressure, and the crude product was separated and purified by column chromatography (n-hexane / ethyl acetate) to obtain 400 mg of a yellow oil with a yield of 81.6%.

[0130] Step 2: Synthesis of 1-(5-(2-fluorophenyl)-1-((3-(3-methylbut-1-yn-1-yl)phenyl)sulfonyl)-1H-pyrrol-3-yl)-N-methylmethanamine (II-5)

[0131] At room temperature, compound 5-1 (400 mg, 0.783 mmol) was dissolved in dichloromethane (4 mL), 4M hydrochloric acid / 1,4-dioxane (2 mL) was added, and the mixture was reacted at room temperature for 1 hour. The mixture was concentrated under reduced pressure, water (5 mL) was added, pH=7-8 was adjusted with saturated NaHCO3 solution, and the mixture was extracted with ethyl acetate (20 mL). The organic phase was washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was separated and purified by column chromatography (DCM / MeOH) to obtain 100 mg of a yellow oil with a yield of 31.2%.

[0132] 1 HNMR: (400MHz, CDCl3) δ7.51-7.49(m, 1H), 7.42-7.30(m, 3H), 7.29-7.28(m, 1H), 7.26-7.24(m, 1H), 7.17-7.1 0 (m, 2H), 7.07-7.03 (m, 1H), 6.22 (s, 1H), 3.62 (s, 2H), 2.80-2.73 (m, 1H), 2.45 (s, 3H), 1.26 (d, J=6.8Hz, 6H).

[0133] Example 6

[0134] Synthesis of 4-(3-((2-(2-fluorophenyl)-4-((methylamino)methyl)-1H-pyrrol-1-yl)sulfonyl)phenyl)-2-methylbut-3-yn-2-ol (Ⅱ-6)

[0135]

[0136] Step 1: Synthesis of tert-butyl ((5-(2-fluorophenyl)-1-((3-(3-hydroxy-3-methylbut-1-yn-1-yl)phenyl)sulfonyl)-1H-pyrrol-3-yl)methyl)(methyl)carbamate (6-1)

[0137] Compound Int 1 (210 mg, 0.4 mmol), compound 3-1 (68 mg, 0.8 mmol), Pd(PPh3)2Cl2 (28 mg, 0.04 mmol), CuI (16 mg, 0.08 mmol), TEA (122 mg, 1.2 mmol), DMF (3 mL) were added to the sealed tube, stirred, replaced with nitrogen 3 times, heated to 80 ° C and stirred for 4 h. TLC spot plate monitoring showed that the reaction was complete and new spots were generated (PE / EA = 3: 1, Rf = 0.3). 30 mL of water was added, and ethyl acetate (30 mL × 2) was used for extraction. The organic phases were combined and washed once with saturated brine (30 mL). The organic phase was dried over anhydrous sodium sulfate, concentrated under reduced pressure at 45 ° C, and separated by column chromatography (PE / EA = 3: 1) to obtain 200 mg of colorless oil.

[0138] Step 2: Synthesis of 4-(3-((2-(2-fluorophenyl)-4-((methylamino)methyl)-1H-pyrrol-1-yl)sulfonyl)phenyl)-2-methylbut-3-yn-2-ol (II-6)

[0139] At room temperature, compound 6-1 (200 mg) and dichloromethane (4 mL) were added to the reaction bottle, stirred to dissolve, 4M hydrochloric acid / 1,4-dioxane (2 mL) was added, and the reaction was carried out at room temperature for 1 h. The reaction was completed and a new spot was generated (DCM / MeOH=10:1, Rf=0.2) by TLC spot plate monitoring. Sodium bicarbonate aqueous solution was added to adjust the pH to 8, and ethyl acetate (20 mL×2) was used for extraction. The organic phases were combined and washed with saturated brine (20 mL×2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated and separated by TLC plate to obtain 50 mg of brown solid.

[0140] 1 HNMR: (400MHz, CD3OD) δ7.69 (s, 1H), 7.68-7.63 (m, 1H), 7.55-7.44 (m, 3H), 7.33 (s, 1H), 7. 21(t, J=8.0Hz, 1H), 7.16-7.06(m, 2H), 6.39(s, 1H), 3.95(s, 2H), 2.61(s, 3H), 1.59(s, 6H).

[0141] Example 7

[0142] Synthesis of 1-(5-(2-fluorophenyl)-1-((3-(3-methoxyprop-1-en-1-yl)phenyl)sulfonyl)-1H-pyrrol-3-yl)-N-methylmethanamine (Ⅱ-7)

[0143]

[0144] Step 1: Synthesis of tert-butyl ((5-(2-fluorophenyl)-1-((3-(3-methoxyprop-1-en-1-yl)phenyl)sulfonyl)-1H-pyrrol-3-yl)methyl)(methyl)carbamate (7-1)

[0145] At room temperature, compound 1-2 (500 mg, 0.98 mmol) was dissolved in ethyl acetate (8 mL), and then Pd / C (50 mg) was added. Under H2 system, the mixture was stirred at room temperature overnight. After the reaction was completed, it was filtered, the filtrate was concentrated, and 400 mg of colorless oily compound was obtained by reverse phase preparation purification with a yield of 79.68%.

[0146] Step 2: Synthesis of 1-(5-(2-fluorophenyl)-1-((3-(3-methoxyprop-1-en-1-yl)phenyl)sulfonyl)-1H-pyrrol-3-yl)-N-methylmethanamine (II-7)

[0147] At room temperature, compound 7-1 (400 mg, 0.78 mmol) was dissolved in dichloromethane (5 mL) solution, 4M hydrochloric acid / 1,4-dioxane (5 mL) was added, and stirred for two hours. After the reaction was complete, it was concentrated and purified by reverse phase preparation to obtain 92 mg of yellow oily compound with a yield of 28.57% and a purity of >95%. [M+H] + :415.1.

[0148] 1 HNMR: (400MHz, CDCl3) δ7.49 (s, 1H), 7.40-7.33 (m, 4H), 7.23 (s, 1H), 7.16-7.09 (m, 2H), 7.01 (t, J=9.2Hz, 1H), 6.4 8 (d, J=11.6Hz, 1H), 6.34 (s, 1H), 5.96-5.92 (m, 1H), 4.03 (d, J=6.4Hz, 2H), 3.75 (s, 2H), 3.35 (s, 3H), 2.50 (s, 3H).

[0149] Example 8

[0150] Synthesis of (E)-1-(1-((3-(2-cyclopropylvinyl)phenyl)sulfonyl)-5-(2-fluorophenyl)-1H-pyrrol-3-yl)-N-methylmethanamine hydrochloride (II-8)

[0151]

[0152] Step 1: Synthesis of tert-butyl (E)-((1-((3-(2-cyclopropylvinyl)phenyl)sulfonyl)-5-(2-fluorophenyl)-1H-pyrrol-3-yl)methyl)(methyl)carbamate (8-2)

[0153] Compound Int 1 (300 mg, 0.57 mmol), 8-1 (223 mg, 1.15 mmol), Pd(dppf)Cl2 (42 mg, 0.06 mmol) and potassium carbonate (198 mg, 1.44 mmol) were dissolved in a mixture of 1,4-dioxane and water (10 / 2 mL) and stirred at 100° C. overnight under nitrogen protection. After the reaction was complete, ethyl acetate (20 mL) was added for dilution, and the mixture was washed with saturated brine (20 mL×2). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. 250 mg of a yellow oily compound was obtained by column chromatography (PE / EA=3:1) with a yield of 85.62%.

[0154] Step 2: Synthesis of (E)-1-(1-((3-(2-cyclopropylvinyl)phenyl)sulfonyl)-5-(2-fluorophenyl)-1H-pyrrol-3-yl)-N-methylmethanamine hydrochloride (II-8)

[0155] At room temperature, compound 8-2 (250 mg, 0.49 mmol) was dissolved in dichloromethane (3 mL) solution, 4M hydrochloric acid / 1,4-dioxane (3 mL) was added, and stirred for two hours. After the reaction was complete, 140 mg of brown solid compound was obtained by filtration, with a yield of 70.01%, a purity of >95%, [M+H] + :411.1.

[0156] 1 HNMR: (400MHz, CD3OD) δ7.77 (s, 1H), 7.59 (d, J=7.6Hz, 1H), 7.52-7.48 (m, 1H), 7.38 (t, J=8.0Hz, 1H), 7.32-7.29 (m, 1H), 7.26 (s, 1H), 7.17 (t, J=7.6Hz, 1H), 7. 12-7.06 (m, 2H), 6.40 (d, J=15.6Hz, 1H), 6.39 (s, 1H), 5.76 (q, J=9.2Hz, 1H), 4.1 0(s, 2H), 2.70(s, 3H), 1.63-1.59(m, 1H), 0.92-0.87(m, 2H), 0.60-0.56(m, 2H).

[0157] Example 9

[0158] Synthesis of (E)-3-(3-((2-(2-fluorophenyl)-4-((methylamino)methyl)-1H-pyrrol-1-yl)sulfonyl)phenyl)-N-methylacrylamide 2,2,2-trifluoroacetate (II-9)

[0159]

[0160] Step 1: Synthesis of ethyl (E)-3-(3-((4-(((tert-butoxycarbonyl)(methyl)amino)methyl)-2-(2-fluorophenyl)-1H-pyrrol-1-yl)sulfonyl)phenyl)acrylate (9-2)

[0161] Compound Int 1 (200 mg, 0.38 mmol), 9-1 (90 mg, 0.57 mmol), Pd(dppf)Cl2 (28 mg, 0.04 mmol) and potassium carbonate (132 mg, 0.96 mmol) were dissolved in a solution of dioxane and water (10 / 2 mL), and stirred at 100° C. overnight under nitrogen protection. After the reaction was complete, ethyl acetate (20 mL) was added for dilution, and the mixture was washed twice with saturated brine (20 mL×2). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. Purification by column chromatography (PE / EA=1:1) gave 180 mg of a yellow oily compound with a yield of 86.96%.

[0162] Step 2: Synthesis of (E)-3-(3-((4-(((tert-butoxycarbonyl)(methyl)amino)methyl)-2-(2-fluorophenyl)-1H-pyrrol-1-yl)sulfonyl)phenyl)acrylic acid (9-3)

[0163] At room temperature, compound 9-2 (500 mg, 0.92 mmol) was dissolved in methanol (10 mL) solution, and then LiOH (45 mg, 1.84 mmol) aqueous solution (5 mL) was added and stirred at room temperature overnight. The methanol solvent was removed by concentration, and the solution pH was adjusted to 6, and extracted with dichloromethane (20 mL×3). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain 450 mg of a yellow solid with a yield of 95.14%, [M+H] + :515.2.

[0164] Step 3: Synthesis of tert-butyl (E)-((5-(2-fluorophenyl)-1-((3-(3-(methylamino)-3-oxoprop-1-en-1-yl)phenyl)sulfonyl)-1H-pyrrol-3-yl)methyl)(methyl)carbamate (9-5)

[0165] At room temperature, compound 9-3 (450 mg, 0.87 mmol), 9-4 (90 mg, 1.32 mmol), HATU (399 mg, 1.05 mmol) and DIEA (337 mg, 2.61 mmol) were dissolved in DMF (6 mL) solution and stirred at room temperature overnight. Water (20 mL) was added, and the mixture was extracted with ethyl acetate (15 mL × 2). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure and purified by column chromatography (ethyl acetate) to obtain 300 mg of a yellow solid with a yield of 65.22%, [M+H]+ :528.2.

[0166] Step 4: Synthesis of (E)-3-(3-((2-(2-fluorophenyl)-4-((methylamino)methyl)-1H-pyrrol-1-yl)sulfonyl)phenyl)-N-methylacrylamide 2,2,2-trifluoroacetate (II-9)

[0167] At room temperature, compound 9-5 (250 mg, 0.47 mmol) was dissolved in dichloromethane (2 mL) solution, and then 4M hydrochloric acid / 1,4-dioxane (2 mL) was added and stirred at room temperature for 2 h. Water (5 mL) was added to dilute, pH was adjusted to 8, and dichloromethane (15 mL×2) was used for extraction. The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure and purified by reverse phase preparation (5-100% acetonitrile / water (0.1% TFA)) to obtain 60 mg of a white solid with a yield of 23.44%, [M+H] + :428.2.

[0168] 1 HNMR: (400MHz, CD3OD) δ7.87 (d, J=8.0Hz, 1H), 7.80 (s, 1H), 7.56-7.48 (m, 4H), 7.42 (d, J=15.6Hz, 1H), 7.18 (t , J=7.6Hz, 1H), 7.11-7.06 (m, 2H), 6.54 (d, J=16.0Hz, 1H), 6.40 (s, 1H), 4.11 (s, 2H), 2.88 (s, 3H), 2.73 (s, 3H).

[0169] Example 10

[0170] Synthesis of 3-(3-((2-(2-fluorophenyl)-4-((methylamino)methyl)-1H-pyrrol-1-yl)sulfonyl)phenyl)-N-methylprop-2-yn-1-amine (Ⅱ-10)

[0171]

[0172] Step 1: Synthesis of tert-butyl ((5-(2-fluorophenyl)-1-((3-(3-(methylamino)prop-1-yn-1-yl)phenyl)sulfonyl)-1H-pyrrol-3-yl)methyl)(methyl)carbamate (10-1)

[0173] At room temperature, compound Int 1 (200 mg, 0.382 mmol) was dissolved in THF (4 mL), and N-methylprop-2-yn-1-amine (106 mg, 1.53 mmol), Pd(PPh3)2Cl2 (27 mg, 0.038 mmol), CuI (15 mg, 0.076 mmol) and triethylamine (0.5 mL) were added in sequence. After N2 replacement, the reaction solution was sealed, heated to 80 ° C and stirred overnight. After the reaction was complete, the solvent was concentrated, and column chromatography was separated and purified (petroleum ether / ethyl acetate = 10: 1-1: 1) to obtain 150 mg of a yellow solid with a yield of 76.70%, [M+H] + :512.7.

[0174] Step 2: Synthesis of 3-(3-((2-(2-fluorophenyl)-4-((methylamino)methyl)-1H-pyrrol-1-yl)sulfonyl)phenyl)-N-methylprop-2-yn-1-amine (II-10)

[0175] At room temperature, compound 10-1 (140 mg, 0.274 mmol) was dissolved in 1,4-dioxane (3 mL), and 4M hydrochloric acid / 1,4-dioxane (3 mL) was added. After reacting at room temperature for 1 hour, the solvent was concentrated, and the crude product was purified by slurrying with n-hexane / ethyl acetate to obtain 100 mg of a gray solid with a yield of 81.39%, [M+H] + :412.6.

[0176] 1 HNMR: (400MHz, CD3OD) δ7.80-7.77(m, 2H), 7.54-7.51(m, 3H), 7.48(s, 1H), 7.22-7.18(m, 1H), 7.11-7.07 (m, 2H), 6.42 (d, J=2.0Hz, 1H), 4.20 (s, 2H), 4.11 (s, 2H), 2.83 (s, 3H), 2.71 (s, 3H).

[0177] Embodiment 11

[0178] Synthesis of 1-(5-(2,4-difluorophenyl)-1-((3-(3-methoxyprop-1-yn-1-yl)phenyl)sulfonyl)-1H-pyrrol-3-yl)-N-methylmethanamine (Ⅱ-11)

[0179]

[0180] Step 1: Synthesis of tert-butyl ((5-(2,4-difluorophenyl)-1-((3-(3-methoxyprop-1-yn-1-yl)phenyl)sulfonyl)-1H-pyrrol-3-yl)methyl)(methyl)carbamate (11-1)

[0181] Compound Int 3 (20 mg, 0.37 mmol), 1-1 (78 mg, 1.11 mmol), bistriphenylphosphine palladium dichloride (26 mg, 0.04 mmol), cuprous iodide (7 mg, 0.04 mmol) and triethylamine (187 mg, 1.85 mmol) were dissolved in DMF (4 mL). The mixture was reacted overnight at 80 ° C in a sealed tube. After the reaction was completed, water (10 mL) was added, and ethyl acetate (15 mL × 2) was extracted. The combined organic phases were washed with saturated brine (20 mL × 1), and the organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. Purification by column chromatography (PE / EA = 3: 1) gave 150 mg of a yellow oily compound with a yield of 76.92%.

[0182] Step 2: Synthesis of 1-(5-(2,4-difluorophenyl)-1-((3-(3-methoxyprop-1-yn-1-yl)phenyl)sulfonyl)-1H-pyrrol-3-yl)-N-methylmethanamine (II-11)

[0183] At room temperature, compound 11-1 (150 mg, 0.28 mmol) was dissolved in dichloromethane (2 mL), and then 4M hydrochloric acid / 1,4-dioxane (2 mL) was added and stirred at room temperature for 2 h. The mixture was concentrated under reduced pressure to obtain 75 mg of a white solid with a yield of 57.25%.

[0184] 1 HNMR: (400MHz, DMSO-d6) δ8.76 (br, 2H), 7.83-7.80 (m, 2H), 7.61 (t, J=8.0Hz, 1H), 7.49 (d, J=8.0Hz, 1H), 7.39 (s , 1H), 7.28 (t, J=8.0Hz, 1H), 7.15-7.13 (m, 2H), 6.49 (s, 1H), 4.36 (s, 2H), 4.01 (s, 2H), 3.36 (s, 3H), 2.55 (s, 3H).

[0185] Example 12

[0186] Synthesis of 1-(1-((3-(cyclopropylethynyl)phenyl)sulfonyl)-5-(2-fluorophenyl)-1H-pyrrol-3-yl)-N-methylmethanamine hydrochloride (Ⅱ-12)

[0187]

[0188] Step 1: Synthesis of tert-butyl ((1-((3-(cyclopropylethynyl)phenyl)sulfonyl)-5-(2-fluorophenyl)-1H-pyrrol-3-yl)methyl)(methyl)carbamate (12-1)

[0189] At room temperature, compound Int 1 (200 mg, 0.382 mmol) was dissolved in DMF (2 mL), and Pd(PPh3)2Cl2 (27 mg, 0.038 mmol), CuI (8 mg, 0.038 mmol), triethylamine (0.5 mL, 3.59 mmol), 4-1 (102 mg, 1.54 mmol) were added respectively. After N2 replacement, the tube was sealed and heated to 80 ° C and stirred overnight. The mixture was concentrated under reduced pressure, and the crude product was separated and purified by column chromatography (n-hexane / ethyl acetate) to obtain 80 mg of a yellow oil with a yield of 41.2%.

[0190] Step 2: Synthesis of (1-((3-(cyclopropylethynyl)phenyl)sulfonyl)-5-(2-fluorophenyl)-1H-pyrrol-3-yl)-N-methylmethanamine hydrochloride (II-12)

[0191] At room temperature, compound 12-1 (80 mg, 0.197 mmol) was dissolved in 4M hydrochloric acid / 1,4-dioxane (2 mL) and reacted at room temperature for 1 hour. The mixture was concentrated under reduced pressure and the crude product was slurried with DCM / MeOH to obtain 30 mg of a white solid with a yield of 34.1%.

[0192] 1 HNMR: (400MHz, CDCl3) δ9.82 (s, 2H), 7.66 (s, 1H), 7.49-7.47 (m, 1H), 7.41-7.36 (m, 1H), 7.31-7.24 (m, 3H), 7.11-7. 05 (m, 2H), 6.99 (t, J=8.8Hz, 1H), 6.59 (s, 1H), 3.99 (s, 2H), 1.46-1.40 (m, 1H), 0.93-0.86 (m, 2H), 0.83-0.79 (m, 2H).

[0193] Example 13

[0194] Synthesis of Ethyl (E)-3-(3-((2-(2-fluorophenyl)-4-((methylamino)methyl)-1H-pyrrol-1-yl)sulfonyl)phenyl)acrylate (II-13)

[0195] At room temperature, compound 9-2 (180 mg, 0.33 mmol) was dissolved in dichloromethane (3 mL) solution, 4M hydrochloric acid / 1,4-dioxane (3 mL) was added, and stirred for two hours. After the reaction was complete, 140 mg of pink solid compound was obtained by filtration, with a yield of 95.89%, a purity of >95%, and [M+H] + :443.1.

[0196] 1HNMR: (400MHz, DMSO-d6) δ8.94 (s, 2H), 8.11 (d, J=8.0Hz, 1H), 7.82 (s, 1H), 7.72 (s, 1H), 7.65-7.59 (m, 2H), 7.55-7.44 (m, 2H), 7.24-7.78 ( m, 2H), 7.06 (t, J=6.0Hz, 1H), 6.64 (d, J=16.0Hz, 1H), 6.53 (s, 1H), 4.22 (q, J=6.8Hz, 2H), 3.97 (s, 2H), 3.35 (s, 3H), 1.28 (t, J=7.2Hz, 3H).

[0197] Embodiment 14

[0198] Synthesis of 1-(5-(2-fluorophenyl)-1-((3-(3-methoxyprop-1-yn-1-yl)-5-(3-methoxypropoxy)phenyl)sulfonyl)-1H-pyrrol-3-yl)-N-methylmethanamine (Ⅱ-14)

[0199]

[0200] Step 1: Synthesis of tert-butyl ((5-(2-fluorophenyl)-1-((3-(3-methoxyprop-1-yn-1-yl)-5-(3-methoxypropoxy)phenyl)sulfonyl)-1H-pyrrol-3)yl)methyl)(methyl)carbamate (14-1)

[0201] At room temperature, compound Int 4 (200 mg, 327.05 μmol), 1-1 (68.77 mg, 981.16 μmol), Pd(PPh3)2Cl2 (11.86 mg, 32.71 μmol), cuprous iodide (6.23 mg, 32.71 μmol) and triethylamine (99.28 mg, 981.16 μmol) were dissolved in tetrahydrofuran (5 mL) solution and reacted overnight at 80 ° C under nitrogen protection. After the reaction was complete, water (8 mL) was added and extracted with ethyl acetate (5 mL×2). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure and purified by column chromatography (PE / EA=3:1) to obtain 70 mg of a yellow viscous solid with a yield of 36.63%, [M+H] + :601.2.

[0202] Step 2: Synthesis of 1-(5-(2-fluorophenyl)-1-((3-(3-methoxyprop-1-yn-1-yl)-5-(3-methoxypropoxy)phenyl)sulfonyl)-1H-pyrrol-3-yl)-N-methylmethanamine (II-14)

[0203] At room temperature, compound 14-1 (80 mg, 151.91 μmol), 4M hydrochloric acid / 1,4-dioxane (1 mL), and dichloromethane (2 mL) were added to a 10 mL single-mouth bottle. The reaction was stirred at room temperature for 2 h. After the reaction was complete, the solvent was removed by concentration under reduced pressure. After adding water (2 mL) to dissolve, the pH was adjusted to 9 with saturated sodium bicarbonate solution. Ethyl acetate (3 mL × 2), the organic phases were combined, washed once with saturated brine (5 mL), dried over anhydrous sodium sulfate, filtered, and dried to obtain 20 mg of an oily substance with a yield of 35.27%, [M+H] + :487.2.

[0204] 1 HNMR: (400 MHz, CDCl3) δ 7.50 (s, 1H), 7.40-7.35 (m, 1H), 7.14-7.08 (m, 3H), 7.04-6.99 (m, 2H), 6.82 (s, 1H), 6.43 (s, 1H), 4.27 (s, 2H), 3.92 (t, J = 6.4 Hz, 2H), 3.86 (s, 2H), 3.50 (t, J = 6 Hz, 2H), 3.43 (s, 3H), 3.32 (s, 3H), 2.53 (s, 3H), 2.09-1.94 (m, 2H). Example 15

[0205] Synthesis of (E)-3-(5-((2-(2-fluorophenyl)-4-((methylamino)methyl)-1H-pyrrol-1-yl)sulfonyl)pyridin-3-yl)-N-methylacrylamide hydrochloride (II-15)

[0206]

[0207] Step 1: Synthesis of ethyl (E)-3-(5-((4-(((tert-butoxycarbonyl)(methyl)amino)methyl)-2-(2-fluorophenyl)-1H-pyrrol-1-yl)sulfonyl)pyridine-3-acrylate (15-2)

[0208] At room temperature, compound Int 2 (500 mg, 0.95 mmol), 9-1 (180 mg, 1.24 mmol), Pd (dppf) Cl2 (63 mg, 0.095 mmol), Na2CO3 (170 mg, 1.9 mmol) were added to 1,4-dioxane (10 mL), water (2 mL) was added, nitrogen was replaced three times, and the temperature was raised to 95 ° C under nitrogen protection, and stirred overnight. After the reaction was completed, the reaction system was concentrated under reduced pressure, and column chromatography was separated and purified (PE / EA = 2: 1) to obtain 170 mg of an oily substance with a yield of 31.95%, [M+H] + :488.2.

[0209] Step 2: Synthesis of (E)-3-(5-((4-(((tert-butoxycarbonyl)(methyl)amino)methyl)-2-(2-fluorophenyl)-1H-pyrrol-1-yl)sulfonyl)pyridin-3-yl)acrylic acid (15-3)

[0210] At room temperature, NaOH (35 mg, 0.88 mmol) was added to 2 mL of water, stirred to dissolve, ethanol (8 mL) was added, compound 15-2 (120 mg, 0.22 mmol) was added, and stirred at room temperature for 30 min. After the reaction was completed, dilute hydrochloric acid was added to adjust the pH to 7, the reaction system was concentrated under reduced pressure, ethyl acetate (15 mL × 2) was added for extraction, and the organic phase was concentrated under reduced pressure to obtain 100 mg of a yellow oil with a yield of 88.49%, [M+H] + :460.2.

[0211] Step 3: Synthesis of tert-butyl (E)-((5-(2-fluorophenyl)-1-((5-(3-(methylamino)-3-oxoprop-1-en-1-yl)pyridin-3-yl)sulfonyl)-1H-pyrrol-3-yl)methyl)(methyl)carbamate (15-4)

[0212] At room temperature, compound 15-3 (100 mg, 0.19 mmol) was added to 5 mL of DMF, HATU (88.5 mg, 0.23 mmol) was added, and the mixture was stirred at room temperature for 30 min. Methylamine hydrochloride (26.2 mg, 0.39 mmol) and DIPEA (100 mg, 0.78 mmol) were added. After the reaction was completed, 15 mL of ethyl acetate and 10 mL of water were added, and the organic phase was separated. The aqueous phase was treated with ethyl acetate (15 mL×2). The organic phases were combined, and the reaction system was concentrated under reduced pressure. Column chromatography (PE / EA=2:1) ​​was performed to obtain 60 mg of a yellow oil with a yield of 59.74%. [M+H] + :473.2.

[0213] Step 4: Synthesis of (E)-3-(5-((2-(2-fluorophenyl)-4-((methylamino)methyl)-1H-pyrrol-1-yl)sulfonyl)pyridin-3-yl)-N-methylacrylamide hydrochloride (II-15)

[0214] At room temperature, compound 15-4 (60 mg, 0.11 mmol) was dissolved in dichloromethane (5 mL) solution, and then 4M hydrochloric acid / 1,4-dioxane (1.0 mL) was added and stirred at room temperature for 2 h. Concentration gave 40 mg of a brown compound with a yield of 75.80%, [M+H] + :429.2.

[0215] 1HNMR: (400MHz, CD3OD) δ9.00 (s, 1H), 8.57 (s, 1H), 7.90 (s, 2H), 7.56-7.51 (m, 1H), 7.47 (s, 1H), 7.23 (t, J=7.6, 1H), 7 .15 (t, J=7.6, 1H), 7.10 (t, J=9.2Hz, 1H), 6.68 (d, J=15.8Hz, 1H), 6.47 (s, 1H), 4.14 (s, 2H), 2.89 (s, 3H), 2.74 (s, 3H).

[0216] H + / K + -ATPase biological evaluation

[0217] The following in vitro screening assay was used to determine the effects of the compounds of the present invention on H + / K + -Inhibition of ATPase activity.

[0218] Experimental materials and instruments:

[0219] ATP, malachite green, valinomycin, ammonium molybdate

[0220] No K + Buffer: 50 mM Tris-HCl pH 6.5, 5 mM magnesium chloride, 10 μM valinomycin

[0221] Contains K + Buffer: 50 mM Tris-HCl pH 6.5, 5 mM magnesium chloride, 10 μM valinomycin, 20 mM KCl MLG colorimetric solution: 0.1% w / v malachite green, 1.5% w / v ammonium molybdate, 0.2% v / v Tween-20 rabbit gastric mucosal microsomes (rich in H + / K +-ATPase), the extraction method is sucrose gradient centrifugation: the rabbit stomach is washed with tap water and 3MNaCl solution respectively, and then the surface water is removed with filter paper. Pre-cooled homogenization buffer (4ml / g tissue) is added, and homogenization is carried out in a tissue homogenizer for 2-5 minutes. After homogenization, if there are larger tissue particles, they can be removed by centrifugation (600g, 10min), and then the supernatant is transferred to a clean centrifuge tube. After centrifugation at 20000g for 30min, the supernatant is transferred to a clean centrifuge tube, and further centrifuged at 100000g for 90min, and the precipitate is collected; the precipitate is suspended with the homogenate, blown evenly, and the protein concentration is measured by Bradford method, and the concentration is adjusted to 10mg / ml; 7.5% Ficoll layering solution is added in equal proportions, and after centrifugation at 100000g for 60min, the middle layer (H + / K + -ATPaseenriched gastric membranes) in a clean centrifuge tube, dilute 4-5 times with homogenate, continue centrifugation at 100000g for 90 min, collect the precipitate; suspend the precipitate with homogenate, homogenize evenly, measure the protein concentration with Bradford method, adjust the concentration to 22.5 mg / ml, and freeze at -80℃ for later use.

[0222] Experimental process:

[0223] 45 μL buffer (containing K + 5 μL of gastric mucosal microsomes (H + / K + -ATPase), then add 5 μL of compound solution, then add 5 μL of 5 mM ATP to start the reaction, pre-react at 37°C for 30 min. Add 15 μL of malachite green solution to terminate the reaction, balance at room temperature for 20 min, and read the absorbance value at 620 nm.

[0224] At the same time, the reaction with the same volume but without KCl was performed as background and subtracted when calculating the enzyme activity.

[0225] Compound IC 50 The values ​​were calculated by the inhibition rate at different concentrations. + / K + -ATPase has obvious inhibitory activity, IC 50The concentration of the active ingredient is 20 to 100 nM, preferably 20 to 50 nM. The control group 1 is selected from Vonoprazan, the preparation method of which is based on patent CN101300229A; the control group 2 is selected from the following structural formula.

[0226]

[0227] IC of some compounds of the present invention 50 Value (H + / K + ATP) are shown in Table 1 below:

[0228] Table 1

[0229]

[0230] Conclusion: The compounds of the present invention have an + / K + -ATPase has obvious inhibitory activity.

[0231] In vitro cytotoxicity assay

[0232] Using Dulbecco's modified Eagle's medium (DMEM; Invitrogen), the cell line HepG2 derived from human liver cancer was cultured and passaged at 5% and 37°C. Cells in the logarithmic growth phase were collected, counted, and resuspended with complete medium. The cell concentration was adjusted to the appropriate concentration (determined according to the results of the cell density optimization test), inoculated into a 96-well plate, and 75μl / well of cell suspension was added according to the following platemap. The test compound was diluted to the corresponding action concentration set with the culture medium, and added to the cells according to the platemap 25μl / well. The action concentration of the test compound started from 100μM, with a 4-fold gradient dilution, a total of 9 concentrations, and 2 replicates. The cells were placed in a 37°C, 100% relative humidity, 5% CO2 incubator and incubated for 24h. 50μL / well CellTiter Glo RT was added and incubated in the dark for 30min. After gentle shaking, the detection was performed in Envision and the inhibition rate was calculated.

[0233] The inhibition rate of the drug on the growth of each cell was calculated according to the following formula: Cell growth inhibition rate % = (1-As / Ac)×100.

[0234] As: OA of sample (cells + CTG + test compound),

[0235] Ac: OA of normal growing cell control (cells+CTG+DMSO).

[0236] IC was calculated using the software Graphpad Prism 6 and the formula XY-analysis / Nonlinearregression (curve fit) / Dose response-Inhibition / log (inhibitor) vs. response-Variable slope (four parameters). 50 Curve fitting and calculation of IC 50 The IC value of the compound of the present invention was found to be 50 Greater than 20μM, with low cytotoxicity.

[0237] The results of in vitro cytotoxicity tests of some compounds of the present invention are shown in the following table (Table 2):

[0238] Table 2:

[0239] serial number <![CDATA[IC 50 ]]> II-2 +++ II-7 +++ II-9 +++ II-11 +++ Control group 1 +++

[0240] Note: IC 50 >20μM is +++, 20μM>IC 50 >10μM is ++, 10μM>IC 50 is +.

[0241] Liver microparticle stability test

[0242] Prepare liver microsomes of the desired species (e.g., mouse, rat, dog, monkey, or human). Prepare 10 mM sample and positive control stock solutions using DMSO as the diluent. Then dilute all stock solutions to a working concentration of 0.25 mM with 70% acetonitrile. The cofactor used in this study is the NADPH regeneration system, which consists of 6.5 mM NADP, 16.5 mM G-6-P, and 3 U / mL G-6-PD. The quenching reagent is an acetonitrile solution containing tolbutamide and propranolol.

[0243] The buffer used in this study was 100 mM potassium phosphate buffer. A mixture containing 0.2 mg / mL liver microsomal protein and 1 μM test article / positive control was incubated in 100 mM potassium phosphate buffer.

[0244] 80 μL of each incubation solution was added to 300 μL of quenching reagent to precipitate protein and prepare a 0-minute sample. After vortexing the sample, 20 μL of NADPH regeneration system was added. 520 μL of each incubation solution was added to 130 μL of NADPH regeneration system to initiate the reaction. The final incubation conditions for 650 μL were: 0.2 mg / mL microsomal protein, 1 μM band / positive control, 1.3 mM NADP, 3.3 mM glucose 6-phosphate, 0.6 U / mL glucose 6-phosphate dehydrogenase. The mixture was placed in a 37°C water bath and gently shaken. 100 μL of the mixture was taken at 0, 5, 10, 30, and 60 minutes, respectively, placed on a 96-well plate containing 300 μL of quenching reagent to precipitate protein, and centrifuged (5000×g, 10 minutes). 80 μL of the supernatant was added to a 96-well detection plate pre-added with 160 μL of ultrapure water and analyzed by LC-MS / MS. Data processing to obtain the elimination half-life (T 1 / 2 , T 1 / 2 =0.693 / K) and in vitro clearance (Cl int ).

[0245] The stability results of some compounds of the present invention in SD rat liver microsomes are shown in the following table (Table 3):

[0246] Table 3:

[0247]

[0248] The experiment found that the compound of the present invention in liver microsomes T 1 / 2 (min)>80min, preferably T 1 / 2 (min)>100min, with good liver microparticle metabolic stability.

[0249] Pharmacokinetic experiments

[0250] The compound under study is administered orally or intravenously (solvent 5% DMSO + 10% Solutol (HS-15) + 85% saline) to animals (such as mice, rats, dogs or monkeys) once, and blood is collected at fixed time points. Immediately after blood sampling, gently invert the test tube at least 5 times to ensure adequate mixing and then place on ice. The blood is anticoagulated with heparin and then centrifuged at 8000pm for 5 minutes to separate the serum from the red blood cells. The serum is aspirated with a pipette and transferred to a 2mL polypropylene tube, labeled with the name of the compound and the time point, and stored in a -40°C refrigerator before LC-MS analysis for testing. High-concentration samples are diluted with blank plasma for determination. After sample processing, the substances in the plasma are quantitatively analyzed by LCMS / MS. The plasma concentration / time curve obtained in this way is used to calculate pharmacokinetic parameters using a validated pharmacokinetic computer program. The experiment found that the compounds of the present invention all have good pharmacokinetic properties.

[0251] After oral administration of the drugs in the groups shown in Table 4 to male SD rats (each group was administered with an equimolar dose, the solvent was 5% DMSO + 10% Solutol (HS-15) + 85% saline, 3 rats in each group), blood samples were collected at fixed time points for testing. The pharmacokinetic parameters of the prototype free molecules of some compounds of the present invention in rat plasma are shown in Table 4 below;

[0252] Table 4:

[0253]

[0254]

[0255] As can be seen from Table 4, the experiment found that the AUC of the prototype free molecule of the compound of the present invention in rat plasma had no significant difference compared with the AUC of the prototype free molecule in plasma of the control group, but the half-life was significantly prolonged, and it had good pharmacokinetic properties.

[0256] Study on the function of hERG potassium channel

[0257] Test system

[0258] Cells: Chinese hamster ovary (CHO) cell line, CHO-hERG cells were used in this experiment.

[0259] Cell culture medium and culture conditions: The complete culture medium is F12 medium supplemented with 10% fetal bovine serum and 1% Selective antibiotics (G418), 89 μg / mL hygromycin B (HB). The recovery medium is F12 medium supplemented with 10% fetal bovine serum. CHO-hERG cells are grown in a high humidity incubator at 37°C (±2°C) and 5% CO2 (4% to 8%). Cells are revived with recovery medium and passaged with complete medium. Cells used for patch clamp experiments are replaced with recovery medium at the last passage.

[0260] Extracellular and intracellular fluid components:

[0261] Reagents External solution (mM) Internal fluid (mM) CaCl2 2 5.37 MgCl2 1 1.75 KCl 4 120 NaCl 145 - Glucose 10 - HEPES 10 10 EGTA - 5 Na2ATP - 4 pH 7.3-7.4 7.2-7.3

[0262] Test methods

[0263] (1) Collect CHO-hERG cells in the exponential growth phase and resuspend them in ECS for later use.

[0264] (2) Manual patch clamp test

[0265] hERG currents were recorded under whole-cell patch clamp technique at room temperature. The output signal of the patch clamp amplifier was converted to digital and filtered at 2.9 kHz. Data were collected using Patchmaster Pro software.

[0266] The cells were placed in a cell recording tank on the stage of an inverted microscope, and a cell in the recording tank was randomly selected for testing. The perfusion system was fixed on the stage of an inverted microscope and the cells were continuously perfused with ECS.

[0267] Manual patch clamp recording microelectrodes were prepared using capillary glass tubes filled with intracellular fluid. On the day of the patch clamp experiment, electrodes were prepared using borosilicate glass tubes (BF150-117-10, SUTTER INSTRUMENT USA). The resistance of the electrodes after filling with ICS was between 2 and 5 MΩ.

[0268] The clamping voltage is -80mV. The first step is to depolarize to +60mV and maintain for 850ms to open the hERG channel. Then, the voltage is set to -50mV and maintained for 1275ms to generate a rebound current or tail current. The peak value of the tail current will be measured and used for analysis. Finally, the voltage is restored to the clamping voltage (-80mV). During the experiment, this command voltage program is repeated every 15s. The manual patch clamp hERG current test command voltage program is as follows Figure 1 shown.

[0269] At the beginning of recording of the perfusion of the solvent control working solution, monitor the tail current peak until more than 3 scanning curves are stabilized, then the test sample / positive control working solution to be tested can be perfused until the inhibitory effect of the test sample / positive control working solution on the hERG current peak reaches a stable state. Generally, the basic coincidence of the peak values ​​of the three most recent consecutive current curves is used as the criterion for judging whether it is in a stable state. After reaching a stable state, continue to perfuse the next concentration of the test sample. One or more test samples / positive controls, or multiple concentrations of the same drug, can be tested on one cell. Different test samples / positive controls need to be rinsed with solvent control working solution until the hERG current returns to more than 80% of the size before the addition of the drug. The standard deviation of the inhibition rate of each recorded cell at the same concentration shall not exceed 15%.

[0270] The positive control cisapride was tested at a concentration of 0.1 μM, and the cells were assayed in duplicate. According to scientific literature reports, 0.1 μM cisapride inhibits hERG current by more than 50%. (Milnes, JT, et al.).

[0271] (3) Manual patch clamp data acceptance criteria

[0272] Sealing standard: After the whole-cell mode is formed, the clamping voltage (-80mV) is applied, and the cell membrane related parameters (Cm, Rm and Ra) can be recorded. A good whole-cell recording should meet the following conditions: path resistance (Rs) is less than 10MΩ; membrane resistance (Rm) is greater than 500MΩ and membrane capacitance (Cm) is less than 100pF.

[0273] Current size: The peak current amplitude before the test article / positive control is between 400pA and 5000pA. Otherwise, the cell is discarded.

[0274] Leakage current: At a clamp voltage of -80mV, the absolute value of the leakage current should be less than 200pA. The current amplitude will be corrected using the leakage current at -80mV. Scans with leakage currents greater than 200pA cannot be used for analysis.

[0275] Data analysis

[0276] For each cell, the inhibition percentage of each concentration of the test article and positive control was calculated from the recorded current response using the following formula: (1-peak tail current recorded after perfusion of the test article / positive control / peak tail current (starting current) recorded after perfusion of the vehicle control) × 100%.

[0277] For each concentration, all cell inhibition percentages were recorded and averaged, IC 50 The values ​​were obtained from the concentration-effect curves by the Hill fitting method.

[0278] Test results

[0279] The results of the inhibition of hERG current by some compounds of the present invention are shown in the following table (Table 5)

[0280] Table 5:

[0281] Compound No. <![CDATA[hERG IC 50 ]]> II-1 +++ II-7 ++ II-9 ++ II-10 +++ II-11 ++ II-12 +++ II-13 + Control group 1 +

[0282] Note: IC 50 >20μM is +++, 20μM>IC 50 >10μM is ++, 10μM>IC 50 >1μM is +.

[0283] The experiment found that the compound of the present invention has a higher hERG IC 50 This indicates that the compound of the present invention has a low risk of cardiotoxicity.

[0284] The preferred embodiments of the present invention are described in detail above. However, the present invention is not limited to the specific details in the above embodiments. Within the technical concept of the present invention, a variety of simple modifications can be made to the technical solution of the present invention, and these simple modifications all belong to the protection scope of the present invention.

[0285] It should also be noted that the various specific technical features described in the above specific embodiments can be combined in any suitable manner without contradiction. In order to avoid unnecessary repetition, the present invention will not further describe various possible combinations.

Claims

1. A pyrrolsulfonyl derivative and a pharmaceutically acceptable salt thereof, characterized in that: The general structural formula of the pyrrole sulfonyl derivatives is shown in formula (I): （I） in: A is phenyl; Dashed lines represent optional bonds; R 1 Selected from C 3-12 Cycloalkyl, C 1-6 Alkyl-NH-C 1-6 Alkyl, C 1-6 Alkyl-OC 1-6 Alkyl, -C(=O)NHR a ; R 2 Selected from H; R 3 is selected from halogen; R 4 Selected from C 1-6 wherein the alkyl group may be optionally further substituted with 1 to 3 deuterium or halogen; n is selected from 0, 1, 2, 3; R a Independently selected from C 1-6 Alkyl, C 1-6 Alkoxy, C 3-7 of a cycloalkyl group.

2. The pyrrole sulfonyl derivative and the pharmaceutically acceptable salt thereof according to claim 1, characterized in that: The structure of the pyrrole sulfonyl derivative is shown in formula (II): （Ⅱ）。 3. The pyrrole sulfonyl derivative and the pharmaceutically acceptable salt thereof according to claim 1, characterized in that: The pyrrole sulfonyl derivative is selected from any one of the following structures: 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 。 4. A pharmaceutical composition, characterized in that The pharmaceutical composition comprises the pyrrole sulfonyl derivative and a pharmaceutically acceptable salt thereof as described in any one of claims 1 to 3 and a pharmaceutically acceptable carrier and / or excipient.

5. The pyrrole sulfonyl derivatives according to any one of claims 1 to 3, the pharmaceutically acceptable salts thereof or the pharmaceutical composition according to claim 4 in the preparation of a gastric acid secretion inhibitor, H + / K + - Use in ATPase inhibitors or potassium ion competitive acid blockers.

6. Use of the pyrrole sulfonyl derivative according to any one of claims 1 to 3, a pharmaceutically acceptable salt thereof, or the pharmaceutical composition according to claim 4 in the preparation of a medicament for treating or preventing the following diseases: peptic ulcer, Zollinger-Ellison syndrome, erosive esophagitis, reflux esophagitis, symptomatic gastroesophageal reflux disease, Barrett's esophagitis, functional dyspepsia, Helicobacter pylori infection, gastric cancer, gastric MALT lymphoma, ulcer caused by nonsteroidal anti-inflammatory drugs, hyperacidity caused by postoperative stress, or ulcer caused by postoperative stress.

7. Use of the pyrrole sulfonyl derivative according to any one of claims 1 to 3, a pharmaceutically acceptable salt thereof, or the pharmaceutical composition according to claim 4 in the preparation of a medicament for inhibiting upper gastrointestinal bleeding caused by peptic ulcer, acute stress ulcer, hemorrhagic gastritis or invasive stress.

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