CN111704613A - Imidazole derivatives and their use as TRPV4 inhibitors - Google Patents
Imidazole derivatives and their use as TRPV4 inhibitors Download PDFInfo
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- C07D471/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00 in which the condensed system contains two hetero rings
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Abstract
The invention relates to imidazole derivatives of formula (I) and pharmaceutically acceptable salts, isomers, prodrugs and pharmaceutical compositions thereof, which are inhibitors of vanilloid 4 transient receptor potential channel (TRPV 4). The invention also discloses a preparation method of the compound and application of the compound as a medicine, in particular application of the compound as a medicine for treating acute lung injury.
Description
Technical Field
The invention relates to a compound capable of inhibiting vanilloid 4 type transient receptor potential channel, pharmaceutically acceptable salts, isomers, prodrugs and pharmaceutical compositions thereof, and also relates to a preparation method thereof and an application thereof as a medicament, in particular to an application thereof as a medicament for acute lung injury.
Background
Transient receptor potential channels (TRP channels) are an important class of cation channels located on cell membranes. It was first discovered in the visual system of drosophila, and mutant drosophila produce only transient, rather than sustained, spikes to sustained light stimuli, hence the name transient receptor potential channel. Many TRP channels have been discovered in many organisms such as drosophila, helminths and mammals. In mammals, 28 TRP channel subtypes have been found. The 28 mammalian TRP channels now found are divided into 6 subfamilies based on amino acid sequence homology: TRPC, TRPV, TRPM, TRPA, TRPP and TRPML. The TRP channel is used as an important receptor of cells, transmits information inside and outside the cells, and is also regulated by messenger molecules and compounds from inside and outside the cells and changes of temperature, osmotic pressure and the like. The different TRP channel regulatory mechanisms differ and many TRP channels can be regulated by multiple stimuli.
TRP channels are widely distributed, in addition to the central nervous system, in the peripheral nervous system, the skin, the cardiovascular system, the respiratory system, the gastrointestinal system, the urogenital system, the immune and endocrine systems, and the like. And unlike the classical voltage-dependent cation channels which participate in relatively specialized functions, the TRP channels are involved in multiple functions such as heat sensation, pain sensation and airway regulation; mediating Ca2+Influx of cells and maintenance of cellsIon homeostasis, regulating vascular permeability and tone; participate in cell growth regulation and the like.
The TRP channel consists of 6 transmembrane (S1-S6) domains, with an intracellular N-and C-terminus. The fragment between S5 and S6 of the TRP channel is embedded in and constitutes an ion passage channel. Whereas the S4 fragment lacks the positively charged amino acid residues normally present in the S4 fragment of the voltage-dependent cation channel, so that the TRP channel is voltage-independent. Furthermore, there are several ankyrin-like repeat structures at the N-terminus of many TRP subtypes. Some TRP isoforms contain a highly conserved TRP domain at the C-terminus, consisting of 25 amino acids with the "EWKFAR" or "VWKYQR" TRP cassette, and some isoforms also have a kinase or phosphorylase domain at the C-terminus.
TRPV4 is one of the members of the TRPV subfamily, also known as VR-OAC, OTRPC4, TRP12 or VRL-2, the first cloning and expression in heterologous systems was based on homology to nematode OSM-9. The TRPV4 channel chromosome is located at 12q 24.1. The human TRPV4 is a homotetramer, each monomer consisting of 871 amino acids. The TRPV4 channel is expressed in nerve, heart, liver, kidney, lung, spleen, testis and other tissues. TRPV4 is activated by hypotonic stress, shear stress, hyperthermia, arachidonic acid and its metabolites, and artificially synthesized small molecules such as GSK1016790A, causing intracellular Ca2+The concentration is increased, which in turn leads to various physiological and pathological processes. Studies have shown that TRPV4 expression is elevated in acute lung injury models, leading to uncontrolled inflammatory responses in the lungs. Whereas TRPV4 inhibitors effectively reduce the inflammatory response. Therefore, it is of great interest to study specific and highly active inhibitors of TRPV 4.
In recent years, there have been some reports in the literature of substances that can inhibit TRPV4 activity, such as: ruthenium red, citral, berberine, and various selective small molecules such as RN-1734, RN-9893, HC-067047, GSK-205, etc. Furthermore, small molecule compounds as TRPV4 inhibitors are described in WO2009111680, WO2009146177, WO2010011912, WO2011091407, WO2011119693, WO2012144661, WO2012174342, WO2013012500, WO2013152109, WO2014008477, WO2016028325, WO2017177200, US 2011130400. The compounds have high in vitro activity in inhibiting TRPV 4.
The TRPV4 inhibitor with the structural general formula is developed according to the characteristics of medicines and the requirements of clinical medicines.
Disclosure of Invention
The compound used in the invention is a potent TRPV4 inhibitor and can be used as a medicament for treating acute lung injury. The compound has an imidazole structure-containing compound as described in claim 1, and the novel imidazole derivative is characterized in that the 1-position of the imidazole ring is substituted by an azetidine or piperidine derivative, and the 2-position is substituted by an alkyl or alkylamino group; in addition, 4 and 5 positions of the imidazole ring are combined with a benzene ring or a hexatomic aromatic heterocycle to form a condensed ring aromatic hydrocarbon structure.
The invention provides a compound shown as a formula I and pharmaceutically acceptable salts, isomers and solvates thereof.
Wherein:
ring T is phenyl and 5-6 membered heteroaryl,
R1is cyano, halogen, AR3、AR3R4，
R2Is AR3，
A is-H, -O, -S, N, -SO2、-C(＝O)、-SO2N、-CO2、-C(＝O)N，
R3Is H, C1～C6The linear or branched alkyl group of (a),
R4is H, C1～C6The linear or branched alkyl group of (a),
x is a substituted or unsubstituted cycloalkyl group,
y is
a) A substituted or unsubstituted aryl group, or a substituted or unsubstituted aryl group,
b) a substituted or unsubstituted heterocyclic group,
r is selected from the following groups:
the formula I according to the invention can also be in the form of its salts, generally with organic or inorganic bases or acids.
Physiologically acceptable salts are preferred in the present invention. Physiologically acceptable salts of the compounds of the invention can be salts of the substances of the invention with inorganic acids, carboxylic acids or sulfonic acids, particularly preferably with hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, nitric acid, perchloric acid, fumaric acid, acetic acid, propionic acid, succinic acid, glycolic acid, formic acid, lactic acid, maleic acid, tartaric acid, citric acid, pamoic acid, malonic acid, hydroxymaleic acid, phenylacetic acid, glutamic acid, benzoic acid, salicylic acid, fumaric acid, p-toluenesulfonic acid, methanesulfonic acid, ethanesulfonic acid, naphthalene-2-sulfonic acid, benzenesulfonic acid, hydroxynaphthoic acid, hydroiodic acid, malic acid, tannic acid, for example. Other acids, such as oxalic, while not in themselves pharmaceutically acceptable, may be used to prepare salts useful as intermediates to obtain the compounds of the invention and pharmaceutically acceptable salts thereof.
The compounds of the invention may exist in tautomeric forms and the invention likewise encompasses such forms.
The compounds of the present invention may also be possible solvates thereof.
The alkyl group is usually a branched or branched alkyl group having 1 to 6, preferably 1 to 4, carbon atoms, particularly preferably 1 to 3 carbon atoms, and preferable examples thereof are methyl, ethyl, n-propyl, isopropyl, tert-butyl, n-pentyl and n-hexyl.
Halogen (halogen for the purposes of the present invention) is fluorine, chlorine, bromine and iodine.
The present invention also relates to synthetic methods for preparing the compounds of formula I of the present invention, comprising:
1) phenyl or 5-6 membered heteroarylo 1- (1-Boc piperidin-4-yl) -imidazole (IIa) is reacted with 4M HCl (III) to give the intermediate 4- (phenyl or 5-6 membered heteroaryloimidazol-1-yl) piperidine hydrochloride (IVa).
Wherein: r1And R2Is as defined in claim 1.
2) Phenyl or 5-6 membered heteroarylo 1- (1-Boc azetidin-3-yl) -imidazole (IIb) is reacted with 4M HCl (III) to give the intermediate 3- (phenyl or 5-6 membered heteroaryloimidazol-1-yl) azetidine hydrochloride (IVb).
Wherein: r1And R2Is as defined in claim 1.
3) Reaction of 4- (phenyl or 5-6 membered heteroarylimidazol-1-yl) piperidine hydrochloride (IVa) with 10- (4-bromophenyl) -9, 9-dimethyl-9, 10-dihydroacridine (V) affords the final product 10- (4- (4- (phenyl or 5-6 membered heteroarylimidazol-1-yl) -piperidin-1-yl) phen-1-yl) -9, 9-dimethyl-9, 10-dihydroacridine (Ii).
Wherein: r1And R2Is as defined in claim 1.
4) Reaction of 3- (phenyl or 5-6 membered heteroarylbenzimidazol-1-yl) azetidine hydrochloride (IVb) with 10- (4-bromophenyl) -9, 9-dimethyl-9, 10-dihydroacridine (V) affords the final product 10- (4- (3- (phenyl or 5-6 membered heteroarylbenzimidazol-1-yl) -azetidin-1-yl) phen-1-yl) -9, 9-dimethyl-9, 10-dihydroacridine (Iii).
Wherein: r1And R2Is as defined in claim 1.
The compounds of formula IIa can be prepared by the following reaction scheme:
Compounds of formula IIb may be prepared by the following reaction scheme:
The invention also relates to a medicament comprising at least one compound according to the invention, preferably together with one or more pharmacologically acceptable excipients or carriers, and to the use thereof for the above-mentioned purposes. Pharmaceutically acceptable carriers herein include, but are not limited to: ion exchangers, aluminum oxide, aluminum stearate, lecithin, serum proteins such as human serum albumin, buffer substances such as phosphates, glycerol, sorbic acid, potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids, water, salts or electrolytes, such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinylpyrrolidone, cellulosic substances, polyethylene glycol, sodium carboxymethylcellulose, polyacrylates, beeswax, lanolin.
The active ingredient may have a systemic and/or topical effect and may therefore be administered by a suitable route, such as orally, parenterally, pulmonarily, nasally, sublingually, lingually, buccally, rectally, transdermally, conjunctivally, topically or in the form of an implant.
The active ingredient may also be administered in a form suitable for such administration.
Suitable for oral administration are the known administration forms which deliver the active ingredient rapidly and/or in a modified manner, such as tablets (uncoated or coated, e.g. with enteric or uncoated tablets), capsules, dragees, granules, pellets, powders, emulsions, suspensions and aerosols.
The use of parenteral administration may avoid the absorption step (intravenous, intra-arterial, intracardiac, intraspinal or intralumbar) or involve absorption (intramuscular, subcutaneous, intradermal, transdermal or intraperitoneal). Administration forms suitable for parenteral administration are, in particular, solutions for injection and infusion, suspensions, emulsions, lyophilisates and preparations in the form of sterile powders.
Suitable for other routes of administration are e.g. inhaled (especially powder inhalation, spray), nasal drops/solutions, sprays; tablets or capsules for lingual, sublingual or buccal administration, suppositories, preparations for the ear and eye, vaginal capsules, aqueous suspensions (lotions, shaking mixtures), lipophilic suspensions, ointments, creams, emulsions, pastes, dusting powders or implants, such as stents.
The active ingredient can be converted into the stated administration forms by methods known per se. This can be achieved with suitable pharmaceutical excipients which are inert and non-toxic. These include, in particular, carriers (for example microcrystalline cellulose), solvents (for example liquid polyethylene glycol), emulsifiers (for example sodium lauryl sulfate), dispersants (for example polyvinylpyrrolidone), synthetic and natural biopolymers (for example proteins), stabilizers (for example antioxidants and ascorbic acid), colorants (for example inorganic pigments such as iron oxide) or flavoring and/or taste masking agents. Where appropriate, the active ingredient may be presented in microencapsulated form in one or more of the above-mentioned carriers.
The above-mentioned pharmaceutical preparations may contain, in addition to the compounds of formula I according to the invention, other pharmaceutically active ingredients.
Drawings
Figure 1 titration curve of the inhibitory activity of the compound of example 1 on TRPV 4.
Detailed Description
The following examples are illustrative of preferred embodiments of the present invention and are not intended to limit the invention in any way.
Firstly, synthesis of related intermediates:
1. intermediate 1: 6-chloro-N4- (1-Boc-piperidin-4-yl) -pyrimidine-4, 5-diamine (intermediate of scheme 1)
20g (122mmol) of 4, 6-dichloro-5-aminopyrimidine and 24.4g (122mmol) of 1-Boc-4-aminopiperidine were dissolved in 244mL of t-butanol, 42.4mL of DIPEA was added, the mixture was heated to reflux, stirred for 16 hours, and the solvent was distilled off under reduced pressure, 100mL of dichloromethane and 100mL of saturated brine were added to the residue, the layers were separated, the aqueous layer was extracted with dichloromethane (2 × 80mL), the organic layers were combined, dried over anhydrous magnesium sulfate, and the solvent was distilled off under reduced pressure, and column chromatography was performed with PE: EA: 2: 1 to obtain 10.0g of a white solid (yield 25.0%).1H-NMR(CHLOROFORM-D，400MHz)：8.07(s，1H)，4.17-4.05(m，2H)，2.92(m，3H)，2.10- 2.04(m，4H)，1.46(s，9H)；ESI-MS(m/z)：328.13[M+H]+
2. Key intermediate 2: 6-chloro-8-isopropylamino-9- (1-Boc-piperidin-4-yl) -9H-purine (intermediate of scheme 1)
The apparatus was dried, 9.1g (27.7mmol) 6-chloro-N4- (1-Boc-piperidin-4-yl) -pyrimidine-4, 5-diamine and 1.1g (27.7mmol) NaH (60%) were placed in a 1000mL three-necked flask under nitrogen, and after addition, 181.6mL of anhydrous pyridine was added with stirring, and 4.4mL (31.3mmol) of isopropyl alcohol was slowly added dropwiseIsopropyl thiocyanate, heated to 80 ℃ and stirred for 1 hour, then 20.7g (38.0mmol) of 1-cyclohexyl-2-morpholinoethylcarbodiimide p-toluenesulfonate was added, the temperature was raised to 90 ℃ and stirred for 2 hours to stop the reaction, the solvent was distilled off under reduced pressure, 100mL of dichloromethane and 100mL of saturated saline were added to the residue to separate the layers, the aqueous layer was extracted with dichloromethane (2 × 80mL), the organic layers were combined, dried over anhydrous magnesium sulfate, the solvent was distilled off under reduced pressure, and DCM: THF 20: 1 was separated by column chromatography to obtain 5.7g of a white solid (yield 52.0%).1H-NMR(CHLOROFORM-D，400MHz)：8.39(s， 1H)，4.54(br，1H)，4.47-4.17(m，4H)，2.94-2.77(m，2H)，2.63-2.45(m，2H)， 1.84(d，J＝11.0Hz，2H)，1.50(s，9H)，1.36(d，J＝6.4Hz，6H).；ESI-MS(m/z)：395.17 [M+H]+
3. Intermediate 3: 6-cyano-8-isopropylamino-9- (1-Boc-piperidin-4-yl) -9H-purine (intermediate of scheme 2)
The apparatus was dried by sequentially adding 3g (7.6mmol) of 6-chloro-8-isopropylamino-9- (1-Boc-piperidin-4-yl) -9H-purine, 1.8g (15.2mmol) of zinc cyanide, 7.1g (7.6mmol) of tris (dibenzylideneacetone) dipalladium, 4.4g (7.6mmol) of 1, 1' -bis (diphenylphosphino) ferrocene into a 250mL three-necked flask, stirring under nitrogen, adding 38.2mL of anhydrous DMF, heating to 100 deg.C, stirring for 12 hours, stopping the reaction, filtering with celite to remove solid impurities, adding 300mL of water, extracting with ethyl acetate (4 × 100mL), washing with 50mL of saturated saline, drying without water, distilling off the solvent under reduced pressure, separating with PE: EA at 1: 1 to obtain 2.2g of a white solid (yield 75.0%).1H-NMR(CHLOROFORM-D，400MHz)：8.58(s，1H)， 4.97(br，1H)，4.50-4.18(m，4H)，2.88(t，J＝12.5Hz，2H)，2.56(dt，J＝12.5， 8.1Hz，2H)，1.89-1.80(m，2H)，1.50(s，9H)，1.38(d，J＝6.5Hz，6H)；ESI-MS(m/z)： 386.22[M+H]+
4. Key intermediate 4: n, N-dimethyl-8-isopropylamino-9- (1-Boc-piperidin-4-yl) -9H-purin-6-yl-carboxamide (intermediate of scheme 2)
2.1g (5.4mmol) 6-cyano-8-isopropylamino-9- (1-Boc-piperidin-4-yl) -9H-purine was dissolved in 62.0mL ethanol and 15.4mL deionized water, 2.6g (46.4mmol) potassium hydroxide was added,heating to 80 deg.C, reacting overnight, distilling off solvent under reduced pressure, adding 50mL methanol to the residue, filtering off insoluble matter, washing the insoluble matter with methanol (4 × 20mL), distilling off methanol under reduced pressure, drying, adding into 250mL three-necked flask directly, forming suspension with 41.8mL dichloromethane, adding 0.46mL DIPEA at 0 deg.C, slowly adding 10.4mL T3P dropwise, controlling the temperature of the reaction system below 1 deg.C, stirring at 0 deg.C for 1 hr, slowly adding 13.1mL (26.2mmol) dimethylamine dropwise, controlling the temperature of the reaction system below 1 deg.C, slowly raising to room temperature, reacting for 3 days, quenching with 50mL saturated sodium bicarbonate solution, layering, extracting the aqueous layer with dichloromethane (2 × 50mL), combining the organic layers, drying with anhydrous magnesium sulfate, distilling off solvent under reduced pressure, DCM: methanol: 20: 1, separating to obtain light yellow solid 0.75g (yield 32. ESI-MS (M/z 432.28): M + H25): column chromatography)]+
5. Intermediate 5: 1-Boc-4- (2-amino-4-methanesulfonylanilino) -piperidine (intermediate of scheme 3)
In a 250mL round bottom flask were added 2g (9.1mmol) of 4-fluoro-3-nitrophenylmethylsulfone and 1.5g (14.1mmol) of sodium carbonate, dissolved in 30.4mL of acetonitrile, 1.8g (9.1mmol) of 1-Boc-4-aminopiperidine was added to the flask with stirring, reacted at room temperature for 2 days, and the solvent was distilled off under reduced pressure, 100mL of ethyl acetate and 100mL of saturated saline were added to the residue, and the layers were separated, the aqueous layer was extracted with ethyl acetate (2 × 80mL), the organic layers were combined, dried over anhydrous magnesium sulfate, and the solvent was distilled off under reduced pressure, and PE: EA: 5: 1 column chromatography was performed to obtain 3.2g of a yellow solid (yield 87.8%).1H-NMR(CHLOROFORM-D，400MHz)：8.77(d，J＝2.2Hz，1H)，8.46(d，J＝7.0Hz，1H)，7.88(dd，J＝9.1，2.2Hz，1H)，7.00(d，J＝9.2 Hz，1H)，4.05(br，2H)，3.82-3.69(m，1H)，3.04(d，J＝10.8Hz，5H)，2.07(d， J＝10.7Hz，2H)，1.77-1.50(m，2H)，1.47(s，9H)；ESI-MS(m/z)：344.10[M-tBu+H]+
A100 mL round-bottomed flask was charged with 3.2g (8.0mmol) of 1-Boc-4- (2-nitro-4-methanesulfonylanilino) piperidine, 2.1g (37.9mmol) of iron powder, and 0.21g (4.0mmol) of ammonium chloride, dissolved in 15.0mL of ethanol and 5.0mL of deionized water, heated to 70 ℃ for 6 hours, filtered through celite, and the filtrate was distilled off under reduced pressure. After drying, 2.4g of crude product was obtained (yield 82%) and used as starting material for the next reaction without further purification.
6. Key intermediate 6: 1- (1-Boc piperidin-4-yl) -2-isopropylamino-5-methanesulfonyl-1H-benzimidazole (intermediate of scheme 3)
The apparatus was dried, 2g (5.4mmol) of 1-Boc-4- (2-amino-4-methanesulfonylanilino) -piperidine was dissolved in 10.8mL of anhydrous pyridine in a 100mL three-necked flask, and under nitrogen protection, 0.62mL (5.4mmol) of isopropyl isothiocyanate was slowly added dropwise, the mixture was heated to 80 ℃ and stirred for 2 hours, cooled to room temperature, 1.3g (6.5mmol) of 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride was added, the mixture was stirred for 18 hours, the reaction was stopped, the solvent was distilled off under reduced pressure, 50mL of dichloromethane and 50mL of saturated saline were added to the residue, the layers were separated, the aqueous layer was extracted with dichloromethane (2 × 40mL), the organic layers were combined, dried over anhydrous magnesium sulfate, the solvent was distilled off under reduced pressure, and column chromatography of DCM: THF ═ 20: 1 was carried out to obtain 1.1g of a white solid (yield 47.3%).1H-NMR(CHLOROFORM-D，400MHz)：8.01(d，J＝1.7Hz，1H)，7.59(dd，J＝8.4，1.3Hz，1H)，7.30(d，J＝8.4Hz， 1H)，4.44-4.22(m，4H)，4.11-4.03(m，1H)，3.03(s，3H)，2.96-2.81(m，2H)， 2.36-2.21(m，2H)，1.89(d，J＝11.2Hz，2H)，1.52(s，9H)，1.36(d，J＝6.3Hz， 6H)；ESI-MS(m/z)：427.18[M+H]+
7. Intermediate 7: 1-Boc-4- (2-amino-5-cyanoanilino) -piperidine (intermediate of scheme 3)
A250 mL round-bottom flask was charged with 2g (12.0mmol) of 3-fluoro-4-nitrobenzonitrile and 2g (18.6mmol) of sodium carbonate, dissolved in 40.1mL of acetonitrile, 2.4g (12.0mmol) of 1-Boc-4-aminopiperidine was added to the reaction flask with stirring, reacted at room temperature for 2 days, and the solvent was distilled off under reduced pressure. To the residue were added 100mL of ethyl acetate and 100mL of saturated brine, the layers were separated, the aqueous layer was extracted with ethyl acetate (2 × 80mL), the organic layers were combined, dried over anhydrous magnesium sulfate, the solvent was distilled off under reduced pressure, and column chromatography was performed using PE: EA ═ 10: 1 to give 4.0g of a yellow solid (yield 95.9%). ESI-MS (m/z): 291.11[ M-tBu + H ] +
A100 mL round-bottomed flask was charged with 4g (11.5mmol) of 1-Boc-4- (2-nitro-5-cyanoanilino) piperidine, 3.1g (54.5mmol) of iron powder, and 0.31g (5.8mmol) of ammonium chloride, dissolved in 21.7mL of ethanol and 7.2mL of deionized water, heated to 70 ℃ for 6 hours, filtered through celite, and the filtrate was distilled off under reduced pressure. After drying, 3.5g of crude product was obtained (yield 96.3%) and used as starting material for the next reaction without further purification.
8. Key intermediate 8: 1- (1-Boc piperidin-4-yl) -2-isopropylamino-6-cyano-1H-benzimidazole (intermediate of scheme 3)
The apparatus was dried, 3.5g (10.9mmol) of 1-Boc-4- (2-amino-5-cyanoanilino) piperidine was dissolved in 21.9mL of anhydrous pyridine in a 100mL three-necked flask, and 1.2mL (10.9mmol) of isopropyl isothiocyanate was slowly added dropwise under nitrogen protection, the mixture was heated to 80 ℃ and stirred for 2 hours, and cooled to room temperature, 2.5g (13.1mmol) of 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride was added thereto, the mixture was stirred for 18 hours, the reaction was stopped, the solvent was distilled off under reduced pressure, 100mL of dichloromethane and 100mL of saturated saline were added to the residue, the layers were separated, the aqueous layer was extracted with dichloromethane (2 × 80mL), the organic layers were combined, dried over anhydrous magnesium sulfate, the solvent was distilled off under reduced pressure, and DCM: THF column chromatography was performed at 50: 1 to obtain 2.4g of a pale brown solid (yield 57.0%).1H-NMR(CHLOROFORM-D，400MHz)：7.45(m，2H)，7.37(dd，J＝8.2，1.1Hz，1H)，4.50-4.18(m，4H)，3.99(t，J ＝12.0Hz，1H)，2.87(t，J＝11.8Hz，2H)，2.24(qd，J＝12.7，4.7Hz，2H)，1.87(d， J＝11.4Hz，2H)，1.52(s，9H)，1.35(d，J＝6.3Hz，6H)；ESI-MS(m/z)：384.24[M-H]+
9. Intermediate 9: 6-chloro-N4- (1-Boc-azetidin-3-yl) -pyrimidine-4, 5-diamine (intermediate of scheme 4)
10g (61mmol) of 4, 6-dichloro-5-aminopyrimidine and 10.5g (61mmol) of 1-Boc-3-aminoazetidine were dissolved in 122mL of t-butanol, 21.2mL of DIPEA was added, the mixture was heated to reflux, stirred for 16 hours, and the solvent was distilled off under reduced pressure, 100mL of dichloromethane and 100mL of saturated saline were added to the residue, the layers were separated, the aqueous layer was extracted with dichloromethane (2 × 80mL), the organic layers were combined, dried over anhydrous magnesium sulfate, and the solvent was distilled off under reduced pressure, and column chromatography was performed with PE: EA: 2: 1 to obtain 5.6g of a white solid (yield 30.6%).1H-NMR(CHLOROFORM-D，400MHz)：8.01(s，1H)，5.85(d，J＝5.7Hz，1H)，4.75-4.66(m，1H)，4.35(dd，J＝9.2，7.5Hz，2H)，3.82(dd，J＝9.0，4.5Hz，2H)，1.44(s， 9H)；ESI-MS(m/z)：300.12[M+H]+
10. Key intermediate 10: 6-chloro-8-isopropylamino-9- (1-Boc azetidin-3-yl) -9H-purine (intermediate of scheme 4)
The apparatus was dried, 1.2g (4.1mmol) of 6-chloro-N4- (1-Boc-azetidin-3-yl) -pyrimidine-4, 5-diamine and 0.16g (4.1mmol) NaH (60%) were put in a 100mL three-necked flask under nitrogen protection, and after addition, 27.0mL of anhydrous pyridine was added under stirring, 0.7mL (4.7mmol) of isopropyl isothiocyanate was slowly added dropwise, the mixture was heated to 80 ℃ and stirred for 1 hour, and then 2.4g (5.7mmol) of 1-cyclohexyl-2-morpholinoethylcarbodiimide p-toluenesulfonate was added, the mixture was heated to 90 ℃ and stirred for 2 hours to stop the reaction, and the solvent was distilled off under reduced pressure, 50mL of dichloromethane and 50mL of saturated saline were added to the residue to separate the layers, and then extracted with dichloromethane (2 × mL), the organic layers were combined, dried over anhydrous magnesium sulfate and the solvent was distilled off under reduced pressure, and DCM: THF was separated by column chromatography at 20: 1 to obtain 0.5g of a white solid (yield 33.0%).1H-NMR(CHLOROFORM-D，400MHz)：8.40(s， 1H)，5.36(m，1H)，4.55-4.40(m，3H)，4.31(dd，J＝10.3，4.0Hz，2H)，1.50(s， 9H)，1.37(d，J＝6.5Hz，6H)；ESI-MS(m/z)：367.16[M+H]+
11. Intermediate 11: 1-Boc-3- (2-amino-4-methanesulfonylanilino) -azetidine (intermediate of scheme 5)
In a 250mL round bottom flask were added 2g (9.1mmol) of 4-fluoro-3-nitrophenylmethylsulfone and 1.5g (14.1mmol) of sodium carbonate, dissolved in 30.4mL of acetonitrile, 1.6g (9.1mmol) of 1-Boc-3-aminoazetidine was added to a reaction flask with stirring, reacted at room temperature for 2 days, the solvent was distilled off under reduced pressure, 100mL of ethyl acetate and 100mL of saturated saline were added to the residue, the layers were separated, the aqueous layer was extracted with ethyl acetate (2 × 80mL), the organic layers were combined, anhydrous magnesium sulfate was dried, the solvent was distilled off under reduced pressure, and PE: EA ═ 5: 1 column chromatography was performed to obtain 3.1g of a yellow solid (yield 91.5%).1H-NMR(CHLOROFORM-D，400MHz)：8.80(d，J＝2.2Hz，1H)，8.62(s，1H)，7.94(dd，J＝9.0，2.1Hz，1H)，6.67(d，J＝9.0Hz，1H)，4.47-4.37(m，3H)，3.96-3.87(m，2H)，3.06(s，3H)，1.45(s，9H)；ESI-MS(m/z)： 316.07[M-tBu+H]+，272.08[M-Boc+H]+
A100 mL round bottom flask was charged with 3g (8.1mmol) of 1-Boc-3- (2-nitro-4-methanesulfonylanilino) azetidine, 2.1g (37.9mmol) of iron powder, 0.21g (4.0mmol) of ammonium chloride, dissolved in 15.0mL of ethanol and 5.0mL of deionized water, heated to 70 ℃ for 6 hours, filtered through celite, and the filtrate was evaporated under reduced pressure. After drying, 2.4g of crude product (yield 87.0%) was obtained and used as starting material for the next reaction without further purification.
12. Key intermediate 12: 1- (1-Boc azetidin-3-yl) -2-isopropylamino-5-methanesulfonyl-1H-benzimidazole (intermediate of scheme 5)
The apparatus was dried, 1.5g (4.4mmol) of 1-Boc-3- (2-amino-4-methanesulfonylanilino) azetidine was dissolved in 8.8mL of anhydrous pyridine in a 100mL three-necked flask, stirred under nitrogen, 0.47mL (4.4mmol) of isopropyl isothiocyanate was slowly added dropwise, heated to 80 ℃ and stirred for 2 hours, cooled to room temperature, 1.0g (5.3mmol) of 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride was added, stirred for 18 hours, the reaction was stopped, the solvent was distilled off under reduced pressure, 50mL of dichloromethane and 50mL of saturated saline were added to the residue, the layers were separated, the aqueous layer was extracted with dichloromethane (2 × 40mL), the organic layers were combined, dried over anhydrous magnesium sulfate, the solvent was distilled off under reduced pressure, and DCM: THF column chromatography was performed at 20: 1 to obtain 1.0g of a white solid (yield 55.4%).1H-NMR(CHLOROFORM-D，400MHz)：8.00(s，1H)，7.64(d，J＝8.4Hz，1H)，7.35(d，J ＝6.6Hz，1H)，7.26(s，1H)，5.03(m，1H)，4.52(t，J＝9.1Hz，2H)，4.37(dd，J＝ 10.0，3.8Hz，2H)，4.25(m，1H)，3.04(s，3H)，1.50(s，9H)，1.35(d，J＝5.6Hz， 6H)；ESI-MS(m/z)：409.90[M+H]+
13. Intermediate 13: 1-Boc-3- (2-amino-5-cyanoanilino) -azetidine (intermediate of scheme 6)
A250 mL round-bottom flask was charged with 2g (12.0mmol) of 3-fluoro-4-nitrobenzonitrile and 2g (18.6mmol) of sodium carbonate, dissolved in 40.1mL of acetonitrile, 2.1g (12.0mmol) of 1-Boc-3-aminoazetidine was added to the reaction flask with stirring, reacted at room temperature for 2 days, and the solvent was distilled off under reduced pressure. To the residue were added 100mL of ethyl acetate and 100mL of saturated brine, and the layers were separated, waterThe layers were extracted with ethyl acetate (2 × 80mL), the organic layers were combined, dried over anhydrous magnesium sulfate, and the solvent was evaporated under reduced pressure, and column chromatography was performed with PE: EA of 10: 1 to give 3.7g (yield 96.5%) of a yellow solid.1H-NMR(CHLOROFORM-D，400MHz)：8.30(d， J＝8.7Hz，1H)，8.22(d，J＝4.3Hz，1H)，7.00(dd，J＝8.7，1.5Hz，1H)，6.82(d，J＝1.1Hz，1H)，4.50-4.39(m，2H)，4.33(m，1H)，3.88(dd，J＝9.3，4.5Hz，2H)， 1.46(s，9H)；ESI-MS(m/z)：263.08[M-tBu+H]+，219.09[M-Boc+H]+
A100 mL round bottom flask was charged with 3.7g (11.6mmol) of 1-Boc-3- (2-nitro-5-cyanoanilino) azetidine, 3.1g (54.5mmol) of iron powder, 0.31g (5.8mmol) of ammonium chloride, dissolved in 21.8mL of ethanol and 7.3mL of deionized water, heated to 70 ℃ for 6 hours, filtered through celite, and the filtrate was evaporated under reduced pressure. After drying, 3.0g of crude product was obtained (yield 89.5%) and used as starting material for the next reaction without further purification.
14. Key intermediate 14: 1- (1-Boc azetidin-3-yl) -2-isopropylamino-6-cyano-1H-benzimidazole (intermediate of scheme 6)
The apparatus was dried, 2.5g (8.7mmol) of 1-Boc-3- (2-amino-5-cyanoanilino) azetidine was dissolved in 17.6mL of anhydrous pyridine in a 100mL three-necked flask, stirred under nitrogen, 0.94mL (8.7mmol) of isopropyl isothiocyanate was slowly added dropwise, heated to 80 ℃ and stirred for 2 hours, cooled to room temperature, 2.0g (10.4mmol) of 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride was added, stirred for 18 hours, the reaction was stopped, the solvent was distilled off under reduced pressure, 50mL of dichloromethane and 50mL of saturated saline were added to the residue, the layers were separated, the aqueous layer was extracted with dichloromethane (2 × 40mL), the organic layers were combined, dried over anhydrous magnesium sulfate, the solvent was distilled off under reduced pressure, and DCM: THF column chromatography was performed at 20: 1 to obtain 1.9g of a white solid (yield 61.7%).1H-NMR(CHLOROFORM-D，400MHz)：7.45(m，3H)，4.98-4.87(m，1H)，4.57-4.48(m， 2H)，4.39-4.22(m，3H)，1.51(s，9H)，1.35(d，J＝6.5Hz，6H)；ESI-MS(m/z)：356.21 [M+H]+
Second, synthesis example of target compound:
example 1: 10- (4- (4- (6-N, N-Dimethylcarboxamido-8-isopropylamino-9H-purin-9-yl) -piperidin-1-yl) -phenyl) -9, 9-dimethyl-9, 10-dihydroacridine
51mg (0.14mmol) of 4- (6-N, N-dimethylformamido-8-isopropylamino-9H-purin-9-yl) piperidine hydrochloride, 51mg (0.14mmol) of 10- (4-bromophenyl) -9, 9-dimethyl-9, 10-dihydroacridine, 3.1mg (0.01mmol) of palladium acetate, 8.2mg (0.03mmol) of 2- (di-tert-butylphosphino) biphenyl and 135mg (0.42mmol) of cesium carbonate are dissolved in 1ml of 1, 4-dioxane, the mixture is heated to 100 ℃ under nitrogen protection, the reaction is stopped after stirring for 18 hours, celite is filtered, the filtrate is collected, the solvent is evaporated under reduced pressure, and the product is isolated by column chromatography to obtain 60mg (yield 70.4%).1H-NMR(CHLOROFORM-D，400MHz)：8.62 (s，1H)，7.44(dd，J＝7.6，1.6Hz，2H)，7.20(q，J＝8.9Hz，4H)，6.94(dtd，J＝ 25.4，7.3，1.5Hz，4H)，6.33(dd，J＝8.1，1.1Hz，2H)，4.58(br，1H)，4.39(m，2H)， 3.99(d，J＝12.1Hz，2H)，3.20(s，3H)，3.11-2.97(m，5H)，2.80(dd，J＝22.5， 11.5Hz，2H)，2.01(d，J＝10.6Hz，2H)，1.68(s，6H)，1.32(d，J＝6.5Hz，6H)；ESI-MS(m/z)：615.39[M-H]+
Example 2: 10- (4- (4- (6-chloro-8-isopropylamino-9H-purin-9-yl) -piperidin-1-yl) -phenyl) -9, 9-dimethyl-9, 10-dihydroacridine
The same procedures used in example 1, 45mg (0.14mmol) of 4- (6-chloro-8-isopropylamino-9H-purin-9-yl) piperidine hydrochloride were reacted with 51mg (0.14mmol) of 10- (4-bromophenyl) -9, 9-dimethyl-9, 10-dihydroacridine to give 42mg (yield 53.7%) of a white solid.1H-NMR(d-DMSO，400MHz)：8.27(s，1H)，7.44(dd，J＝7.8，1.5Hz，2H)， 7.36(d，J＝7.5Hz，1H)，7.25(d，J＝9.0Hz，2H)，7.15(d，J＝9.0Hz，2H)，6.99 -6.93(m，2H)，6.89-6.83(m，2H)，6.22(dd，J＝8.2，1.2Hz，2H)，4.54(m，1H)， 4.23-4.14(m，1H)，4.00(d，J＝12.4Hz，2H)，2.96-2.73(m，4H)，1.87(d，J ＝13.3Hz，2H)，1.58(s，6H)，1.27(d，J＝6.6Hz，6H)；ESI-MS(m/z)：578.28[M+H]+
Example 3: 10- (4- (4- (2-isopropylamino-5-methanesulfonyl-1H-benzimidazol-1-yl) -piperidin-1-yl) -phenyl) -9, 9-dimethyl-9, 10-dihydroacridine
The same procedures used in example 1, 80mg (0.21mmol) of 4- (2-isopropylamino-5-methanesulfonyl-1H-benzimidazol-1-yl) piperidine hydrochloride were reacted with 78mg (0.21mmol) of 10- (4-bromophenyl) -9, 9-dimethyl-9, 10-dihydroacridine to give 76mg (yield 57.4%) of a white solid.1H-NMR(d-DMSO，400MHz)：8.01(d，J＝1.7Hz，1H)，7.57(dd， J＝8.4，1.7Hz，1H)，7.38(d，J＝8.7Hz，2H)，7.34(d，J＝8.4Hz，1H)，6.97(d， J＝8.8Hz，2H)，4.41(br，1H)，4.28(dq，J＝13.2，6.5Hz，1H)，4.11(t，J＝12.2 Hz，1H)，3.87(d，J＝12.7Hz，2H)，3.04(s，3H)，2.92(t，J＝11.7Hz，2H)，2.54 (qd，J＝12.4，3.8Hz，2H)，1.98(d，J＝10.3Hz，2H)，1.71(s，6H)，1.35(d，J＝ 6.4Hz，6H)；ESI-MS(m/z)：620.31[M+H]+
Example 4: 10- (4- (4- (2-isopropylamino-6-cyano-1H-benzimidazol-1-yl) -piperidin-1-yl) -phenyl) -9, 9-dimethyl-9, 10-dihydroacridine
The same procedures as in example 1, 65mg (0.20mmol) of 4- (2-isopropylamino-5-cyano-1H-benzimidazol-1-yl) piperidine hydrochloride and 74mg (0.20mmol) of 10- (4-bromophenyl) -9, 9-dimethyl-9, 10-dihydroacridine are reacted to give 50mg (yield 43.6%) of a white solid.1H-NMR(CHLOROFORM-D，400MHz)：7.58(s，1H)，7.44(ddd，J＝15.7，9.6，4.8Hz，4H)，7.23(q，J＝8.7Hz，4H)，6.94(dt，J＝26.9，7.0Hz，4H)，6.33 (d，J＝7.9Hz，2H)，4.38-4.27(m，1H)，4.18-3.94(m，3H)，3.04(t，J＝12.0 Hz，2H)，2.59(dd，J＝21.1，12.1Hz，2H)，2.08(d，J＝11.7Hz，2H)，1.68(s，6H)，1.38(d，J＝6.3Hz，6H)；ESI-MS(m/z)：567.33[M+H]+
Example 5: 10- (4- (4- (2-methyl-1H-imidazo [4, 5-b ] -pyridin-1-yl) -piperidin-1-yl) -phenyl) -9, 9-dimethyl-9, 10-dihydroacridine
The procedure is as in example 1, 64mg (0.25mmol) of 4- (2-methyl-1H-imidazo [4, 5-b)]Pyridin-1-yl) piperidine hydrochloride was reacted with 93mg (0.25mmol) of 10- (4-bromophenyl) -9, 9-dimethyl-9, 10-dihydroacridine to give 70mg (yield 55.6%) of a white solid.1H-NMR(CHLOROFORM-D，400MHz)：8.50(dd，J＝4.8，1.3Hz，1H)，7.84(d，J＝7.7Hz，1H)，7.46(dd，J＝7.6，1.6Hz，2H)，7.24(q，J＝9.0Hz，4H)，7.14 (dd，J＝8.1，4.8Hz，1H)，6.96(dtd，J＝25.5，7.3，1.4Hz，4H)，6.35(dd，J＝8.1， 1.2Hz，2H)，4.45(t，J＝12.2Hz，1H)，4.03(d，J＝12.7Hz，2H)，3.08(t，J＝12.0 Hz，2H)，2.77(s，3H)，2.74-2.61(m，2H)，2.10(dd，J＝12.4，2.4Hz，2H)，1.69 (s，6H)；ESI-MS(m/z)：500.28[M+H]+
Example 6: 10- (4- (3- (6-chloro-8-isopropylamino-9H-purin-9-yl) -azetidin-1-yl) -phenyl) -9, 9-dimethyl-9, 10-dihydroacridine
The procedure is as in example 1, 50mg (0.16mmol) of 3- (6-chloro-8-isopropylamino-9H-purin-9-yl) azetidine hydrochloride with 60mg (0.16mmol) of 10- (4-bromophenyl) -9, 9-dimethyl-9, 10-dihydroacridine to give 35mg (yield 38.6%) of a white solid.1H-NMR(CHLOROFORM-D，400MHz)：8.40(s，1H)，7.70(d，J＝7.6Hz，1H)，7.46(dd，J＝7.6，1.5Hz，2H)，7.28(d，J＝8.6Hz，2H)，7.03-6.84(m，6H)， 6.30(dd，J＝8.1，1.2Hz，2H)，5.63(m，1H)，4.49(m，3H)，4.32(dd，J＝9.8，2.4 Hz，2H)，1.69(s，6H)，1.37(d，J＝6.5Hz，6H)；ESI-MS(m/z)：550.25[M+H]+
Example 7: 10- (4- (3- (2-isopropylamino-5-methanesulfonyl-1H-benzimidazol-1-yl) -azetidin-1-yl) -phenyl) -9, 9-dimethyl-9, 10-dihydroacridine
The same procedures as in example 1, 70mg (0.20mmol) of 3- (2-isopropylamino-5-methanesulfonyl-1H-benzimidazol-1-yl) azetidine hydrochloride and 74mg (0.20mmol) of 10- (4-bromophenyl) -9, 9-dimethyl-9, 10-dihydroacridine were reacted to give 45mg (yield 37.6%) of a white solid.1H-NMR(CHLOROFORM-D，400MHz)：8.00(d，J＝1.8Hz，1H)， 7.59(dd，J＝8.3，1.8Hz，1H)，7.45(dd，J＝7.6，1.6Hz，2H)，7.28-7.22(m，3H)， 6.96(dtd，J＝26.7，7.3，1.4Hz，4H)，6.87-6.73(m，3H)，6.3l(dd，J＝8.1，1.3 Hz，2H)，5.21-5.12(m，1H)，4.49(t，J＝8.7Hz，2H)，4.40(dd，J＝9.4，2.9Hz， 2H)，4.33(dq，J＝13.2，6.6Hz，1H)，3.05(s，3H)，1.69(s，6H)，1.36(d，J＝6.5 Hz，6H)；ESI-MS(m/z)：592.27[M+H]+
Example 8: 10- (4- (3- (2-isopropylamino-6-cyano-1H-benzimidazol-1-yl) -azetidin-1-yl) -phenyl) -9, 9-dimethyl-9, 10-dihydroacridine
The procedure is as in example 1, 60mg (0.20mmol) of 3- (2-isopropylamino-5-cyano-1H-benzimidazol-1-yl) azetidine hydrochloride are reacted with 74mg (0.20mmol) of 10- (4-bromophenyl) -9, 9-dimethyl-9, 10-dihydroacridine to give 60mg (yield 67.6%) of a white solid.1H-NMR(CHLOROFORM-D，400MHz)：7.58-7.40(m，5H)，7.28(d，J＝8.6Hz，2H)，6.97(dddd，J＝14.8，8.7，7.3，1.4Hz，4H)，6.88-6.82(m， 2H)，6.32(dd，J＝8.1，1.3Hz，2H)，5.12(br，1H)，4.52(t，J＝8.6Hz，2H)，4.47 -4.33(m，3H)，1.69(s，6H)，1.38(d，J＝6.5Hz，6H).；ESI-MS(m/z)：539.29[M+H]+
Third, example of biological Activity test
One) measurement method
1. Gene synthesis
The human TRPV4 gene containing a Kozak sequence, Afl II and Xho I restriction sites was synthesized by GENEWIZ and then inserted into the pcDNA5/FRT/TO vector.
2. Development of CHO-TRPV4 Stable cell line
1X106 Flp-In-CHO cells were placed In 5ml complete medium without any antibiotics and cultured overnight In T-25 flasks at 37 ℃ under 5% CO2
Complete medium: ham's F12K + 10% FBS + 1% Penicillin-Streptomyces + 100. mu.g/mLzeocin
On the day of transfection, media and transfection reagents were pre-warmed to RT prior to the experiment
Mu.l Lipofectamine 3000 was diluted in 250. mu.L Opti-MEM I Reduced Serum Medium
Mu.g of DNA (pOG 44: pcDNA5/FRT-TO-TRPV 4: 9: 1) was diluted in 250. mu.L of Opti-MEM IRED Serum Medium, gently mixed, then 16. mu. L P3000 TO the diluted DNA was added, gently mixed
The diluted DNA was gently mixed with the diluted Lipofectamine 3000, and incubated at room temperature for 5-10 minutes
The complex was added to a T-25 flask containing cells and 5mL of fresh whole medium without any antibiotics, mixed gently, and incubated at 37 ℃ under 5% CO2 for 24 hours
24h after transfection, cells were trypsinized, resuspended in zeocin-free complete medium and then all cells were transferred into T-225 flasks. Cell fusion was about 30% after overnight incubation at 37 ℃ under 5% CO2
The medium was changed to selective medium and incubated at 37 ℃ with 5% CO2
Selective culture medium: ham's F-12K + 10% FBS +1X Penicilin-Streptomyces + 800. mu.g/mLHygromycin B.
Culturing transfected CHO-TRPV4 cells with replacement of selection medium every 3-4 days until colony formation (about 10-14 days)
CHO-TRPV4 positive clones will be trypsinized and cultured in selective media, further amplified and cryopreserved
3. Inhibitor IC50Titration function test
Flp-In-CHO TRPV4 stable cells were cultured In selective medium. Maintain cell density near confluency
mu.L of cell seeding medium (Ham's F-12K + 10% FBS) was added to 384-well cell culture plates (Corning, 3712) per well, 6.5K cells/well, incubated overnight at 37 ℃ and 5% CO2
The 20X fraction A was thawed at room temperature, diluted to 2X with 5mM probenecid in buffer (1X HBSS +20mM HEPES) and left at room temperature.
The 384 well plates were removed from the incubator and allowed to equilibrate at room temperature for 10 min. The medium was changed to buffer by an Apricot pipetting device, 20. mu.L of buffer was retained in each well after the last wash step, then 20. mu.l 2X Component A containing 5mM probenecid was added to each well, centrifuged at 200g for 3-5s, and incubated at room temperature for 2h
Starting from 10mM, 3-fold serial dilutions were performed and used as reference or test compounds, 240nL of diluted compounds were transferred to 384-well plates (Corning, 3657) using an Echo pipetting system, centrifuged at 200g for 1min, and then 40. mu.L of buffer solution was added to #3657 plates to form a 6X compound plate. Shaking and mixing at room temperature for 20min
Preparation of 6X GSK1016790A (180nM, 120nM, 60nM and 30nM)
Adding 10 μ L of 6X compound prepared in step 5 to 384-well cell culture plate, centrifuging at 200g for 3-5s, and incubating at RT for 15-30min
10 μ L of GSK1016790A to 384 well cell culture plates prepared in step 6 were added by FLIPR Tetra and data were collected by FLIPR Tetra.
II) measurement results
The compound of example 1 in the synthesis example of the target compound was tested for its inhibitory activity against TRVP4 in vitro according to the method described above, and its inhibitory IC50 value was 205.9nM, which is comparable to the activity of the positive control HC 067047.
Activity assay titration curves are shown in figure 1. titration curves for TRPV4 inhibition activity of the compound of example 1.
Claims (9)
1. A compound shown as a formula I and pharmaceutically acceptable salts, isomers and solvates thereof.
Wherein:
ring T is phenyl and 5-6 membered heteroaryl,
R1is cyano, halogen, AR3、AR3R4，
R2Is AR3，
A is-H, -O, -S, -N, -SO2、-C(＝O)、-SO2N、-CO2、-C(＝O)N，
R3Is H, C1～C6The linear or branched alkyl group of (a),
R4is H, C1～C6The linear or branched alkyl group of (a),
x is a substituted or unsubstituted cycloalkyl group,
y is
a) A substituted or unsubstituted aryl group, or a substituted or unsubstituted aryl group,
b) a substituted or unsubstituted heterocyclic group,
r is selected from the following groups:
2. the compound of claim 1, and pharmaceutically acceptable salts, isomers, solvates thereof. Wherein:
ring T is phenyl and 6-membered heteroaryl,
R1is cyano, -Cl, AR3、AR3R4，
R2Is AR3，
A is-H, -N, -SO2、-C(＝O)N，
R3Is H, C1～C4The linear or branched alkyl group of (a),
R4is H, C1～C4The linear or branched alkyl group of (a),
x is a substituted or unsubstituted cycloalkyl group,
y is
a) A substituted or unsubstituted aryl group, or a substituted or unsubstituted aryl group,
b) substituted or unsubstituted heterocyclyl.
3. The compound of claim 1, and pharmaceutically acceptable salts, isomers, solvates thereof.
Wherein:
ring T is phenyl, pyridyl or pyrimidinyl,
R1is cyano, -Cl, AR3、AR3R4，
R2Is AR3，
A is-H, N, -SO2、-C(＝O)N，
R3Is H, methyl, ethyl or isopropyl,
R4is H, methyl, ethyl orThe isopropyl group is a group selected from the group consisting of isopropyl,
x is piperidine or azetidine,
y is a benzene ring or a pyridine ring.
4. The compound of claim 1, wherein the compound has the structure, and pharmaceutically acceptable salts, isomers, solvates thereof.
5. A process for preparing a compound according to any one of claims 1 to 4, which comprises:
1) phenyl or 5-6 membered heteroarylo 1- (1-Boc piperidin-4-yl) -imidazole (II a) is reacted with 4M HCl (III) to afford intermediate 4- (phenyl or 5-6 membered heteroaryloimidazol-1-yl) piperidine hydrochloride salt (IV a).
Wherein: r1、R2And ring T is as defined in claim 1.
2) Phenyl or 5-6 membered heteroarylo 1- (1-Boc azetidin-3-yl) -imidazole (II b) is reacted with 4M HCl (III) to give the intermediate 3- (phenyl or 5-6 membered heteroaryloimidazol-1-yl) azetidine hydrochloride (IV b).
Wherein: r1、R2And ring T is as defined in claim 1.
3) Reaction of 4- (phenyl or 5-6 membered heteroarylimidazol-1-yl) piperidine hydrochloride (IV a) with 10- (4-bromophenyl) -9, 9-dimethyl-9, 10-dihydroacridine (V) affords the final product 10- (4- (4- (phenyl or 5-6 membered heteroarylimidazol-1-yl) -piperidin-1-yl) phen-1-yl) -9, 9-dimethyl-9, 10-dihydroacridine (I i).
Wherein: r1、R2And ring T is as defined in claim 1.
4) Reaction of 3- (phenyl or 5-6 membered heteroarylbenzimidazol-1-yl) azetidine hydrochloride (IV b) with 10- (4-bromophenyl) -9, 9-dimethyl-9, 10-dihydroacridine (V) affords the final product 10- (4- (3- (phenyl or 5-6 membered heteroarylbenzimidazol-1-yl) -azetidin-1-yl) phen-1-yl) -9, 9-dimethyl-9, 10-dihydroacridine (Iii).
Wherein: r1、R2And ring T is as defined in claim 1.
6. A medicament comprising at least one compound of formula I as defined in claim 1 or a pharmaceutically acceptable salt, isomer, solvate thereof and at least one further excipient.
7. Use of a compound of formula I as defined in claim 1 or a pharmaceutically acceptable salt, isomer, solvate thereof for the manufacture of a medicament for the treatment of acute lung injury.
8. The use of a compound of formula I as defined in claim 1, or a pharmaceutically acceptable salt, isomer, solvate thereof, for the manufacture of a medicament for the treatment of acute respiratory distress syndrome.
9. The use as claimed in any one of claims 7 to 8, wherein the compound which inhibits TRPV4 activity is used with a compound of formula I as defined in claim 1 or a pharmaceutically acceptable salt, isomer, solvate thereof.
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