Fibroblast activating protein inhibitor as well as preparation method and application thereof

Abstract

Disclosed is a compound or a pharmaceutically acceptable salt thereof, the compound comprising a moiety of Formula (I). The invention discloses a preparation method of the compound and application of the compound in preparation of a fibroblast activating protein inhibitor. # imgabs0 #

Description

Technical Field

[0001] The present invention relates to the field of pharmaceutical chemistry, and in particular to a small molecule fibroblast activation protein (FAP) inhibitor, a preparation method and application thereof. Background Art

[0002] Fibroblast activation protein is a type II membrane-bound serine protease with dipeptidyl peptidase activity. It often exists in the form of a dimer and plays a vital role in the metabolism of a variety of endogenous peptides and peptide drugs. It is generally hardly expressed in normal human tissues. Studies have shown that FAP is significantly expressed in diseases such as cellular fibrosis and inflammation, and is particularly highly selectively expressed on the surface of stromal fibroblasts of epithelial malignancies, including breast cancer, gastric cancer, lung cancer, colorectal cancer, and ovarian cancer. Given the widespread expression of FAP in tumors, it has now become a molecular target for tumor imaging and treatment, and can be used in positron emission tomography (PET) imaging and targeted therapy of a variety of malignant tumors.

[0003] At present, fibroblast activation protein inhibitors (FAPI), represented by quinoline carboxylic acid derivatives combined with radionuclide labeling, have been successfully applied in the field of tumor imaging and have made important progress. Reported high-affinity FAP small molecule inhibitors, such as FAPI-02 and FAPI-04, can achieve various types of tumor imaging and do not increase the signal in normal tissue cells, especially in the kidney, liver and thyroid tumor cells with a higher detection rate. However, since FAPI is fast in the blood circulation and can be quickly cleared, it also means that it can be quickly eluted at the tumor site, which is very unfavorable for tumor treatment. Higher doses or frequent administration are required to meet the treatment effect, which increases the probability of adverse reactions and limits its wide application in tumor treatment. Summary of the invention

[0004] In one aspect, the present application provides a compound or a pharmaceutically acceptable salt thereof, in particular a FAP inhibitor, the compound comprising a moiety of formula (I),

[0005]

[0006] in,

[0007] Said Freedom to choose The group composed of.

[0008] In one aspect, there is provided a process for preparing a compound comprising a moiety of formula (I), particularly a compound of formula (II), more particularly a compound of formula (III), or a pharmaceutically acceptable salt thereof, which comprises contacting a compound comprising a moiety of formula (7) or a protected derivative thereof, particularly a compound of formula (7a) or a protected derivative thereof, more particularly a compound of formula (7b) or a protected derivative thereof, with a compound of formula (7x).

[0009]

[0010] In one aspect, there is provided the use of a compound of the present application or a pharmaceutically acceptable salt thereof in the preparation of a fibroblast activation protein inhibitor.

[0011] The compound of the present application or a pharmaceutically acceptable salt thereof, through its moiety of formula (I), exhibits good affinity for the FAP protein and can be used as a small molecule inhibitor of FAP to improve the therapeutic and imaging effects of small molecule inhibitors of FAP, having the potential for clinical application. Detailed Description of the Invention

[0012] To further illustrate the technical means and effects adopted by the present application to achieve the intended purpose, the following, in conjunction with preferred embodiments, details the specific embodiments, structures, features and effects according to the present application.

[0013] Terms and Definitions

[0014] As used herein, the singular terms refer to one or more than one. For example, "element" or "an element" both refer to one element or more than one element. As used herein, the term "plurality" means at least two.

[0015] As used herein, the term "about" means approximate, within the range of about or near. When the term "about" is used in combination with a numerical range, it modifies the range by expanding the boundaries above or below the provided value. Generally, the term "about" as used herein causes the value to vary up and down by 10% from the provided value. In one aspect, the term "about" means plus or minus 20% of the value of the number it modifies. For example, "about 50%" means within the range of 45% - 55%. Numerical ranges mentioned herein by endpoints include all integers and fractions included within that range (e.g., "1 to 5" includes 1, 1.5, 2, 2.75, 3, 3.90, 4 and 5). It should also be understood that all integers and fractions are considered to be modified by the term "about".

[0016] As used herein, the terms "comprising", "including", or "containing", being non-exclusive or open-ended terms, are intended to mean that a combination (such as an apparatus, a composition, a method, etc.) includes the recited elements (such as each unit of the apparatus, each component of the composition, the substantial steps of the method, etc.), but does not exclude other elements. As used herein, the term "consisting essentially of", when used to define compositions and methods, means excluding other elements that have any substantial effect on the combination for the stated purpose, but does not exclude other elements that do not substantially affect the basic and novel features of the present invention. As used herein, the term "consisting of" means excluding other combinations of elements (units, components, substantial steps, etc.), but does not, unless otherwise stated, mean excluding trace amounts of unavoidable impurities. Embodiments defined by each of these connecting terms are within the scope of the present invention. As a specific embodiment thereof, a technical solution disclosed including the terms "comprising", "including", or "containing" should also be regarded as simultaneously disclosing the corresponding technical solutions including the terms "consisting essentially of" and "consisting of".

[0017] As used herein, the term "and / or" means and encompasses any and all possible combinations of one or more of the associated listed items. When used in a list of two or more items, the term "and / or" means that any one of the listed items can be included alone, or any combination of two or more of the listed items can be included. For example, if a group, combination, or composition, etc., is described as including (or containing) components A, B, C, and / or D, then the composition can include A alone; include B alone; include C alone; include D alone; include a combination of A and B; include a combination of A and C; include a combination of A and D; include a combination of B and C; include a combination of B and D; include a combination of C and D; include a combination of A, B, and C; include a combination of A, B, and D; include a combination of A, C, and D; include a combination of B, C, and D; or include a combination of A, B, C, and D.

[0018] As described herein, the term "pharmaceutical composition" means a composition comprising at least one active ingredient, which is acceptable for studying specific, effective results in mammals (such as but not limited to humans). Based on the needs of those skilled in the art, those of ordinary skill in the art will understand and know the techniques suitable for determining whether the active ingredient has the desired effective effect.

[0019] As used herein, the term "pharmaceutically acceptable salt" refers to salts derived from the compounds of the present invention, retaining the biological effectiveness and properties of the compounds of the present invention and generally not being biologically or otherwise undesirable salts. Due to the presence of amino and / or carboxyl groups or groups similar thereto, the compounds of the present invention are capable of forming acid salts and / or base salts. The term "pharmaceutically acceptable salt" may refer to pharmaceutically acceptable addition salts prepared from pharmaceutically acceptable non-toxic acids or bases (including inorganic acids and bases and organic acids and bases).

[0020] The term "protected derivative" means a derivative of a compound (such as a compound specified herein), in which one or more functional groups of the compound (such as functional groups specified herein) are protected or blocked to avoid unwanted reactions. Functional groups that can be protected are, for example, carboxyl groups, amino groups, hydroxyl groups, thiol groups, carbonyl groups, etc., but are not limited thereto. Representative protecting groups for carboxyl groups are, for example, esters (such as p-methoxybenzyl esters), amides, and hydrazides, but are not limited thereto; representative protecting groups for amino groups are, for example, carbamates (such as tert-butoxycarbonyl) and amides, but are not limited thereto; representative protecting groups for hydroxyl groups are, for example, ethers and esters, but are not limited thereto; representative protecting groups for thiol groups are, for example, thioethers and thioesters, but are not limited thereto; representative protecting groups for carbonyl groups are, for example, acetals and ketals, but are not limited thereto.

[0021] Embodiments of the present application

[0022] In one aspect, the present application provides a compound or a pharmaceutically acceptable salt thereof, particularly a FAP inhibitor, the compound comprising a moiety of formula (I),

[0023]

[0024] wherein,

[0025] the is selected from the group consisting of .

[0026] In some embodiments, the compound is a compound of formula (II),

[0027]

[0028] wherein,

[0029] the is selected from the group consisting of and

[0030] the –L is –H, a chelating group or a fluorescent group.

[0031] In some embodiments, the compound is a compound of formula (III),

[0032]

[0033] wherein,

[0034] the is selected from the group consisting of and the group.

[0035] In some embodiments, the chelating group or fluorescent group is selected from 1,4,7,10-tetraazacyclododecane-N,N′,N″,N″′-tetraacetic acid (DOTA), 1,4,7-triazacyclononane-N,N′,N″-triacetic acid (NOTA), diethylenetriamine-N,N,N′,N″,N″-pentaacetic acid (DTPA), 1,4,8,11-tetraazacyclotetradecane-1,4,8,11-tetraacetic acid (TETA), 2,2′-((6-amino-1-(4,7-bis(carboxymethyl)-1,4,7-triazonan-1-yl)hexan-2-yl)azanediyl)diacetic acid (NETA), 1,4,7,10-tetraazacyclododecane-1,4,7-triacetic acid (DO3A), ethylenebis(o-hydroxyphenyl)glycine (EHPG), N,N′-bis(2-hydroxybenzyl)ethylenediamine-N,N′-diacetic acid (HBED), 1,4,7,10-tetraazacyclododecane-α,α′,α″,α″′-tetramethyl-N,N′,N″,N″′-tetraacetic acid (DOTMA), 1,4,8,11-tetraazacyclotetradecane-1,4,8,11-(methyltetraacetic acid)11-(methyltetraacetic acid), TETMA), ethylenediamine tetraacetic acid (EDTA), 1,3-propylenediaminetetraacetic acid (PDTA), triethylenetetraaminehexaacetic acid (TTHA), 1,5,10-N,N′,N″-tris(2,3-dihydroxybenzoyl)-tricatecholate (LICAM), 1,3,5-N,N′,N″-tris(2,3-dihydroxybenzoyl)aminomethylbenzene (MECAM), and 6-hydrazinonicotinic acid (HYNIC), and groups consisting of derivatives or moieties formed by subtracting one or more hydrogens therefrom.

[0036] In some embodiments, L is H, or is selected from the group consisting of 1,4,7,10-tetraazacyclododecane-N,N′,N″,N″′-tetraacetic acid, 1,4,7-triazacyclononane-N,N′,N″-triacetic acid, diethylenetriamine-N,N,N′,N″,N″-pentaacetic acid, 1,4,8,11-tetraazacyclotetradecane-1,4,8,11-tetraacetic acid, 2,2′-((6-amino-1-(4,7-bis(carboxymethyl)-1,4,7-triazanon-1-yl)hexane-2-yl)azanediyl)diacetic acid, 1,4,7,10-tetraazacyclododecane-1,4,7-triacetic acid, ethylenebis(o-hydroxyphenyl)glycine, N,N′-bis(2-hydroxybenzyl)ethylenediamine-N,N′-diacetic acid, 1,4,7,10-tetraazacyclododecane-α,α′,α″,α″′-tetramethyl-N,N′,N″,N″′-tetraacetic acid, 1,4,8,11-tetraazacyclotetradecane-1,4,8,11-(methyltetraacetic acid), ethylenediamine tetraacetic acid, 1,3-propylenediaminetetraacetic acid, triethylenetetraaminehexaacetic acid, 1,5,10-N,N′,N″-tris(2,3-dihydroxybenzoyl)-tricatecholate, 1,3,5-N,N′,N″-tris(2,3-dihydroxybenzoyl)aminomethylbenzene, and 6-hydrazinonicotinic acid, and groups consisting of derivatives or moieties formed by subtracting one or more hydrogens therefrom.

[0037] In some embodiments, L is H.

[0038] In one aspect, there is provided a process for preparing a compound comprising a moiety of formula (I), particularly a compound of formula (II), more particularly a compound of formula (III), or a pharmaceutically acceptable salt thereof, which comprises contacting a compound comprising a moiety of formula (7) or a protected derivative thereof, particularly a compound of formula (7a) or a protected derivative thereof, more particularly a compound of formula (7b) or a protected derivative thereof, with a compound of formula (7x).

[0039]

[0040] In some embodiments, the method further comprises preparing a compound of formula (2) or a protected derivative thereof from a compound of formula (1) or a protected derivative thereof, particularly by contacting a compound of formula (1) or a protected derivative thereof with a compound of formula (1x).

[0041]

[0042] In some embodiments, the method further comprises preparing a compound of formula (3) or a protected derivative thereof from a compound of formula (2) or a protected derivative thereof.

[0043]

[0044] In some embodiments, the method further comprises preparing a compound of formula (4) or a protected derivative thereof from a compound of formula (3) or a protected derivative thereof.

[0045]

[0046] In some embodiments, the method further comprises preparing a compound of formula (5) or a protected derivative thereof from a compound of formula (4) or a protected derivative thereof, particularly by contacting a compound of formula (4) or a protected derivative thereof with a compound of formula (4x) or a protected derivative thereof.

[0047]

[0048] In some embodiments, the method further comprises preparing a compound of formula (6) or a protected derivative thereof from a compound of formula (5) or a protected derivative thereof, particularly by contacting a compound of formula (5) or a protected derivative thereof with a compound of formula (5x).

[0049]

[0050] In some embodiments, the method further comprises preparing a compound comprising a moiety of formula (7) or a protected derivative thereof from a compound of formula (6) or a protected derivative thereof, in particular by contacting a compound of formula (6) or a protected derivative thereof with a compound of formula (6x) or a protected derivative thereof, wherein the is selected from the group consisting of In some embodiments, the method further comprises preparing a compound of formula (7a) or a protected derivative thereof from a compound of formula (6) or a protected derivative thereof, in particular by contacting a compound of formula (6) or a protected derivative thereof with a compound of formula (6xa) or a protected derivative thereof, wherein the is selected from the group consisting of In some embodiments, the method further comprises preparing a compound of formula (7b) or a protected derivative thereof from a compound of formula (6) or a protected derivative thereof, in particular by contacting a compound of formula (6) or a protected derivative thereof with a compound of formula (6xb) or a protected derivative thereof, wherein the is selected from the group consisting of In one aspect, there is provided the use of a compound of the present application or a pharmaceutically acceptable salt thereof in the preparation of a fibroblast activation protein inhibitor. In one aspect, there is provided the use of a compound of the present application or a pharmaceutically acceptable salt thereof as a fibroblast activation protein inhibitor. In one aspect, there is provided a fibroblast activation protein inhibitor comprising or consisting of a compound of the present application or a pharmaceutically acceptable salt thereof. In one aspect, there is provided the use of a compound of the present application or a pharmaceutically acceptable salt thereof in inhibiting fibroblast activation protein. In one aspect, there is provided a method of inhibiting fibroblast activation protein, comprising administering a compound of the present application or a pharmaceutically acceptable salt thereof.

[0051]

[0052] In some embodiments, the fibroblast activation protein inhibitor is a drug targeted at tumors, fibrotic cells, or inflammation, particularly tumors, more particularly stromal fibroblasts of tumors. In some embodiments, the fibroblast activation protein inhibitor is a drug for diagnosing or treating, particularly for radio-diagnosing or treating, particularly for imaging, tumors, fibrotic cells, or inflammation, particularly tumors, more particularly stromal fibroblasts of tumors.

[0053]

[0054] ​In one aspect, there is provided the use of a compound of the present application or a pharmaceutically acceptable salt thereof in the diagnosis or treatment, particularly in the radio-diagnosis or treatment, particularly in imaging, of tumors, fibrotic cells, or inflammation, particularly tumors, more particularly stromal fibroblasts of tumors. In one aspect, there is provided the use of a compound of the present application or a pharmaceutically acceptable salt thereof in the diagnosis or treatment, particularly in imaging, of tumors, fibrotic cells, or inflammation, particularly tumors, more particularly stromal fibroblasts of tumors.

[0055] In one aspect, there is provided a method for targeting tumors, fibrotic cells, or inflammation, particularly tumors, more particularly stromal fibroblasts of tumors, comprising administering a compound of the present application or a pharmaceutically acceptable salt thereof. In one aspect, there is provided a method for diagnosing or treating, particularly in the radio-diagnosis or treatment, particularly in imaging, of tumors, fibrotic cells, or inflammation, particularly tumors, more particularly stromal fibroblasts of tumors, comprising administering a compound of the present application or a pharmaceutically acceptable salt thereof.

[0056] In some embodiments, the tumor is an epithelial-derived tumor. In some embodiments, the tumor is a malignant tumor. In some embodiments, the tumor is breast cancer, gastric cancer, lung cancer, colorectal cancer, or ovarian cancer.

[0057] Examples

[0058] Example 1: Preparation of Compound HX07

[0059] In this example, compound HX07 was prepared according to the following synthetic route.

[0060]

[0061] Step 1: Preparation of 6-methoxyquinoline-2,4-dicarboxylic acid (Compound 2)

[0062] Take a 250 mL three-necked round-bottomed glass bottle, weigh 5-methoxyisatin (2.0 g, 11.3 mmol) and potassium hydroxide (6.3 g, 113 mmol), add them to the reaction flask, measure 50 mL of distilled water with a measuring cylinder and add it to the reaction flask, stir to dissolve, then slowly add pyruvic acid (2.0 g, 11.3 mmol) to the reaction solution with a syringe. Place the reaction flask in an oil bath at 40 °C and react overnight. After the reaction is completed, place the reaction flask in an ice-water bath to cool, and adjust the pH to 3 with concentrated hydrochloric acid. A large amount of yellow solid precipitates. Filter under reduced pressure, and wash the solid 3 times with distilled water. Place the product in a vacuum drying oven to dry to obtain a yellow solid compound 2 (2.3 g, yield 82.4%).

[0063] Step 2: Preparation of 6-methoxyquinoline-4-carboxylic acid (Compound 3)

[0064] Take a 250 mL three-necked round-bottomed glass flask, weigh compound 2 (2.0 g, 8.1 mmol) and place it in the reaction flask. Measure 40 mL of nitrobenzene with a measuring cylinder and transfer it to the reaction flask. Stir to dissolve. Place the reaction flask in a heating jacket and heat it up to 220 °C, and heat the reaction for 6 hours. After the reaction is completed, place the reaction flask in an ice-water bath to cool it down. Add 100 mL of petroleum ether. A large amount of solid precipitates. Filter it under reduced pressure, and wash the solid with petroleum ether 3 times. Place the product in a vacuum drying oven to dry it, and obtain brown solid compound 3 (1.41 g, yield 85.7%).

[0065] Step 3: Preparation of 6-hydroxyquinoline-4-carboxylic acid (Compound 4)

[0066] Take a 100 mL three-necked round-bottomed glass flask, weigh compound 3 (1.5 g, 7.4 mmol) and place it in the reaction flask. Measure 30 mL of 48% aqueous hydrobromic acid solution with a measuring cylinder and transfer it to the reaction flask. Stir to dissolve under nitrogen protection. Place the reaction flask in an oil bath at 130 °C and react overnight. After the reaction is completed, place the reaction flask in an ice-water bath to cool it down. Use 30% sodium hydroxide solution to adjust the pH to 6. A large amount of solid precipitates. Filter it under reduced pressure, and wash the solid with distilled water 3 times. Place the product in a vacuum drying oven to dry it, and obtain brownish-black solid compound 4 (0.98 g, yield 70.1%).

[0067] Step 4: Preparation of tert-butyl (6-hydroxyquinoline-4-carbonyl)glycinate (Compound 5)

[0068] Take a 100 mL three-necked round-bottomed glass flask, weigh compound 4 (600 mg, 3.2 mmol), 2-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate HATU (1.3 g, 3.5 mmol), and N,N-diisopropylethylamine DIPEA (1.1 mL, 6.4 mmol) and place them in the reaction flask. Protect it with nitrogen. Add 10 mL of N,N-dimethylformamide to the reaction flask and stir at room temperature for 0.5 hour. Then dissolve tert-butyl glycinate (42 mg, 3.2 mmol) in 5 mL of N,N-dimethylformamide and transfer it to the reaction flask. React at room temperature overnight. Monitor the reaction progress by TLC plate spotting. After the reaction is complete, stop the reaction. Add saturated ammonium chloride solution to the reaction flask, extract it with dichloromethane (3×20 mL), collect the organic phase, wash it with water 2 - 3 times, collect the organic phase and dry it with anhydrous Na2SO4. Remove the solvent under reduced pressure, and separate and purify it by column chromatography (dichloromethane∶methanol = 20∶1) to obtain yellow oily compound 5 (412 mg, yield 42.6%). LC / MS: 303.3[M+H] + 。

[0069] Step 5: Preparation of tert-butyl (6-(3-chloropropoxy)quinoline-4-carbonyl)glycinate (Compound 6)

[0070] Take a 100 mL three-necked round-bottomed flask, weigh compound 5 (400 mg, 1.3 mmol) and potassium carbonate (366 mg, 2.6 mmol) and place them in the reaction flask. Under nitrogen protection, add 20 mL of N,N-dimethylformamide to the reaction flask. Subsequently, dissolve 1-bromo-3-chloropropane (408 mg, 2.6 mmol) in 5 mL of N,N-dimethylformamide and transfer it to the reaction flask. Place the reaction flask in an oil bath at 60 °C and react overnight. Add saturated ammonium chloride solution to the reaction flask, extract with dichloromethane (3 × 20 mL), collect the organic phase, wash it with water 2 - 3 times, collect the organic phase and dry it with anhydrous Na2SO4. Remove the solvent under reduced pressure and purify by column chromatography (dichloromethane∶methanol = 50∶1) to obtain a yellow oily compound 6 (365 mg, yield 73.2%). LC / MS: 379.6 [M+H] + 。

[0071] Step 6: Preparation of tert-butyl 4-(3-((4-((2-(tert-butoxy)-2-oxoethyl)carbamoyl)quinolin-6-yl)oxy) propyl)piperazine-1-carboxylate (Compound 7)

[0072] Take a 100 mL three-necked round-bottomed flask, weigh compound 6 (300 mg, 0.8 mmol), 1-tert-butoxycarbonylpiperazine (298 mg, 1.6 mmol) and potassium iodide (266 mg, 1.6 mmol) and place them in the reaction flask. Under nitrogen protection, add 30 mL of N,N-dimethylformamide to the reaction flask. Place the reaction flask in an oil bath at 60 °C and react overnight. After the reaction is completed, add saturated ammonium chloride solution to the reaction flask, extract with dichloromethane (3 × 20 mL), collect the organic phase, wash it with water 2 - 3 times, collect the organic phase and dry it. Remove the solvent under reduced pressure and purify by column chromatography (dichloromethane∶methanol = 30∶1) to obtain a yellow oily compound 7 (251 mg, yield 59.4%). LC / MS: 529.7 [M+H] + 。

[0073] Step 7: Preparation of (6-(3-(4-(tert-butoxycarbonyl)piperazin-1-yl)propoxy)quinoline-4-carbonyl)glycine (Compound 8)

[0074] Take a 100 mL three-necked round-bottomed flask, weigh compound 7 (250 mg, 0.47 mmol) and transfer it to the reaction flask. Add 15 mL of tetrahydrofuran to the reaction flask and stir to dissolve. Subsequently, add 5 mL of 20% sodium hydroxide solution to the reaction flask and stir at room temperature overnight. After the reaction is completed, extract with dichloromethane (3 × 20 mL), collect the organic phase, remove the solvent under reduced pressure and purify by column chromatography (dichloromethane∶methanol = 10∶1) to obtain a yellow oily compound 8 (160 mg, yield 72.1%). LC / MS: 473.7 [M+H] + 。

[0075] Step 8: Preparation of tert-butyl 4-(3-((4-((2-((1-cyanocyclopropyl)amino)-2-oxoethyl)carbamoyl)quinolin-6-yl)oxy)propyl)piperazine-1-carboxylate (Compound 9)

[0076] Take a 50 mL three-necked round-bottomed flask, weigh compound 8 (150 mg, 0.32 mmol), 2-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate HATU (182 mg, 0.48 mmol), and N,N-diisopropylethylamine DIPEA (0.11 mL, 0.64 mmol) and place them in the reaction flask. Under nitrogen protection, add 10 mL of N,N-dimethylformamide to the reaction flask and stir at room temperature for 0.5 h. Then dissolve 1-amino-cyclopropyl cyanide hydrochloride (57 mg, 0.48 mmol) in 5 mL of N,N-dimethylformamide and transfer it to the reaction flask. React at room temperature overnight. After the reaction is completed, add saturated ammonium chloride solution to the reaction flask, extract with dichloromethane (3 × 20 mL), collect the organic phase, wash it with water 2 - 3 times, collect the organic phase and dry it, remove the solvent under reduced pressure, and separate and purify by column chromatography (dichloromethane∶methanol = 10∶1) to obtain yellow solid compound 9 (108 mg, yield 62.9%). LC / MS: 537.6 [M + H] + 。

[0077] Step 9: Preparation of N-(2-((1-cyanocyclopropyl)amino)-2-oxoethyl)-6-(3-(piperazin-1-yl)propoxy) quinoline-4-carboxamide (Compound HX07)

[0078] Take a 50 mL three-necked round-bottomed flask, weigh compound 9 (100 mg, 0.19 mmol) and transfer it to the reaction flask. Add 20 mL of ethyl acetate to the reaction flask and stir to dissolve. Then add 4M hydrochloric acid / ethyl acetate solution (0.24 mL, 0.95 mmol) to the reaction flask and stir at room temperature for 5 h. After the reaction is completed, add saturated sodium bicarbonate solution to the reaction flask, extract with dichloromethane (3 × 20 mL), collect the organic phase, wash it with water 2 - 3 times, collect the organic phase and dry it, remove the solvent under reduced pressure, and separate and purify by column chromatography (dichloromethane∶methanol = 8∶1) to obtain yellow solid compound HX07 (41 mg, yield 49.5%). LC / MS: 437.5 [M + H] + 。

[0079] Example 2: Preparation of Compound HX11

[0080] In this example, compound HX11 was prepared according to the following synthetic route.

[0081]

[0082] Step 10: 4-(3-((4-((2-(2-cyanopiperidin-1-yl)-2-oxoethyl)carbamoyl)quinolin-6-yl) oxy)propyl)piperazine-1-carboxylate (Compound 10)

[0083] Take a 50 mL three-necked round-bottomed flask, weigh compound 8 (150 mg, 0.32 mmol), 2-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate HATU (182 mg, 0.48 mmol), and N,N-diisopropylethylamine DIPEA (0.11 mL, 0.64 mmol) and place them in the reaction flask. Under nitrogen protection, add 10 mL of N,N-dimethylformamide to the reaction flask and stir at room temperature for 0.5 hour. Subsequently, dissolve 2-cyanopiperidine hydrochloride (53 mg, 0.48 mmol) in 5 mL of N,N-dimethylformamide and transfer it to the reaction flask. React at room temperature overnight. After the reaction is completed, add saturated ammonium chloride solution to the reaction flask, extract with dichloromethane (3 × 20 mL), collect the organic phase, wash it with water 2 - 3 times, collect the organic phase and dry it, remove the solvent under reduced pressure, and separate and purify by column chromatography (dichloromethane∶methanol = 15∶1) to obtain yellow solid compound 9 (103 mg, yield 57.1%). LC / MS: 565.6 [M + H] + 。

[0084] Step 11: Preparation of N-(2-(2-cyanopiperidin-1-yl)-2-oxoethyl)-6-(3-(piperazin-1-yl)propoxy)quin oline-4-carboxamide (Compound HX11)

[0085] Take a 50 mL three-necked round-bottomed flask, weigh compound 10 (100 mg, 0.18 mmol) and transfer it to the reaction flask. Add 20 mL of ethyl acetate to the reaction flask and stir to dissolve. Subsequently, add 4M hydrochloric acid / ethyl acetate solution (0.23 mL, 0.90 mmol) to the reaction flask and stir at room temperature for 4 hours. After the reaction is completed, add saturated sodium bicarbonate solution to the reaction flask, extract with dichloromethane (3 × 20 mL), collect the organic phase, wash it with water 2 - 3 times, collect the organic phase and dry it, remove the solvent under reduced pressure, and separate and purify by column chromatography (dichloromethane∶methanol = 10∶1) to obtain yellow solid compound HX11 (29 mg, yield 34.7%). LC / MS: 464.5 [M + H] + 。

[0086] Example 3: Preparation of Compound HX16

[0087] In this example, compound HX16 was prepared according to the following synthetic route.

[0088]

[0089] Step 12: 4-(3-((4-((2-(1H-indol-1-yl)-2-oxoethyl)carbamoyl)quinolin-6-yl)oxy propyl)piperazine-1-carboxylate (Compound 11)

[0090] Take a 50 mL three-necked round-bottomed flask, weigh compound 8 (150 mg, 0.32 mmol), 2-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate HATU (182 mg, 0.48 mmol), and N,N-diisopropylethylamine DIPEA (0.11 mL, 0.64 mmol) and place them in the reaction flask. Under nitrogen protection, add 10 mL of N,N-dimethylformamide to the reaction flask and stir at room temperature for 0.5 hour. Subsequently, dissolve indole (56 mg, 0.48 mmol) in 5 mL of N,N-dimethylformamide and transfer it to the reaction flask, and react at room temperature overnight. After the reaction is completed, add saturated ammonium chloride solution to the reaction flask, extract with dichloromethane (3 × 20 mL), collect the organic phase, wash it with water 2 - 3 times, collect the organic phase and dry it, remove the solvent under reduced pressure, and separate and purify by column chromatography (dichloromethane∶methanol = 10∶1) to obtain yellow solid compound 9 (116 mg, yield 63.4%). LC / MS: 572.6 [M+H] + 。

[0091] Step 13: Preparation of N-(2-(1H-indol-1-yl)-2-oxoethyl)-6-(3-(piperazin-1-yl)propoxy)quinoline- 4-carboxamide (Compound HX16)

[0092] Take a 50 mL three-necked round-bottomed flask, weigh compound 11 (100 mg, 0.21 mmol) and transfer it to the reaction flask. Add 20 mL of ethyl acetate to the reaction flask and stir to dissolve. Subsequently, add 4M hydrochloric acid / ethyl acetate solution (0.27 mL, 1.06 mmol) to the reaction flask and stir at room temperature for 5 hours. After the reaction is completed, add saturated sodium bicarbonate solution to the reaction flask, extract with dichloromethane (3 × 20 mL), collect the organic phase, wash it with water 2 - 3 times, collect the organic phase and dry it, remove the solvent under reduced pressure, and separate and purify by column chromatography (dichloromethane∶methanol = 10∶1) to obtain yellow solid compound HX16 (43 mg, yield 43.1%). LC / MS: 471.5 [M+H] + 。

[0093] Biological activity test of FAP inhibitors

[0094] Using the MicroScale Thermophoresis (MST) technique, the affinity between the FAP inhibitor and FAP protein was measured, and the K d value was automatically analyzed and calculated by software to reflect the affinity between the substrate molecule and FAP protein. The smaller the K d value, the higher the affinity.

[0095] The specific experimental steps are as follows.

[0096] Configure buffer solution

[0097] Add 25 μL of Tween-20 to 50 mL of PBS solution and mix well to obtain PBS-T.

[0098] Protein buffer replacement

[0099] The buffer system of FAP protein is tris buffer, so a desalting column is needed to replace the buffer, and the buffer system is replaced with PBS-T buffer system.

[0100] Configure test compound solution

[0101] Dissolve the compound to be tested with DMSO to prepare a solution with a concentration of 50 mM.

[0102] Affinity test

[0103] Dilute FAP protein to 200 nM with PBS-T, and also dilute RED-tris-NTA dye to 5 μM with PBS-T; take 2 μL of the dye (5 μM) and mix it with 98 μL of PBS-T to obtain 100 μL of the dye (100 nM); mix 90 μL of the protein (200 nM) with 90 μL of the dye (100 nM), incubate in the dark for 30 min, after incubation, centrifuge at 4 °C and 15000 g for 10 min, and take the supernatant to a new clean centrifuge tube.

[0104] Dilute the compound with PBS-T, and at the same time prepare a PBS-T buffer containing DMSO, so that the DMSO content in the PBS-T buffer is the same as that in the compound dilution. Prepare two rows of eight-connected tubes, add 10 μL of the prepared PBS-T buffer containing DMSO to tubes 2 - 16, and add 20 μL of the compound solution to be tested to the first tube. Perform serial dilution, take 10 μL of the compound solution to be tested from tube 1 and add it to tube 2, mix well by pipetting repeatedly, then take 10 μL and add it to tube 3 to mix well, and then complete the dilution of tubes 4 - 16 in the same way, and finally remove the excess 10 μL from tube 16. Add 10 μL of the labeled FAP protein (the final concentration of the labeled protein becomes 50 nM) to tubes 1 - 16 respectively and mix well by pipetting, and incubate in the dark at room temperature for 10 min. After sucking the sample with a capillary tube, perform detection on the machine, and calculate the K d value.

[0105] Through the FAP protein affinity test, it can be known that the K dThe values are 33.8 nm, 615 nm, and 649 nm respectively. The results show that HX11 and HX16 molecules have no obvious affinity for FAP protein; while HX07 shows good affinity with FAP protein, and is expected to be used as a small molecule inhibitor of FAP, improving the treatment and imaging effects of small molecule inhibitors of FAP, and having the potential for clinical application.

[0106] The above embodiments are only the preferred embodiments of the present application, and the scope of protection of the present application cannot be limited thereby. Any non-substantial changes and substitutions made by those skilled in the art on the basis of the present application belong to the scope of protection required by the present application.

Claims

1. A compound or a pharmaceutically acceptable salt thereof, said compound comprising a moiety of formula (I), wherein, The said selected from the group consisting of group.

2. The compound or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that, said compound is a compound of formula (II), wherein, The said selected from the group consisting of and said –L is –H, a chelating group or a fluorescent group.

3. The compound or a pharmaceutically acceptable salt thereof according to claim 2, wherein, said L is H, or is selected from the group consisting of 1,4,7,10-tetraazacyclododecane-N,N′,N″,N″′-tetraacetic acid, 1,4,7-triazacyclononane-N,N′,N″-triacetic acid, diethylenetriamine-N,N,N′,N″,N″-pentaacetic acid, 1,4,8,11-tetraazacyclotetradecane-1,4,8,11-tetraacetic acid, 2,2′-((6-amino-1-(4,7-bis(carboxymethyl)-1,4,7-triazanyl)hexane-2-yl)azanediyl)diacetic acid, 1,4,7,10-tetraazacyclododecane-1,4,7-triacetic acid, ethylenebis(o-hydroxyphenyl)glycine, N,N′-bis(2-hydroxybenzyl)ethylenediamine-N,N′-diacetic acid, 1,4,7,10-tetraazacyclododecane-α,α′,α″,α″′-tetramethyl-N,N′,N″,N″′-tetraacetic acid, 1,4,8,11-tetraazacyclotetradecane-1,4,8,11-(methyltetraacetic acid), ethylenediaminetetraacetic acid, 1,3-propanediaminetetraacetic acid, triethylenetetraminehexaacetic acid, 1,5,10-N,N′,N″-tris(2,3-dihydroxybenzoyl)-tris(orthocatechol), 1,3,5-N,N′,N″-tris(2,3-dihydroxybenzoyl)aminomethylbenzene, and 6-hydrazinonicotinic acid, and derivatives or moieties formed by subtracting one or more hydrogens therefrom.

4. The compound or a pharmaceutically acceptable salt thereof according to claim 1, wherein said compound is a compound of formula (III), wherein, The said selected from the group consisting of constitute a group.

5. A process for preparing the compound according to any one of claims 1 to 4 or a pharmaceutically acceptable salt thereof, characterized in that, comprising: contacting a compound comprising a moiety of formula (7) or a protected derivative thereof with a compound of formula (7x).

6. The preparation method according to claim 5, wherein, said method further comprises preparing a compound of formula (2) or a protected derivative thereof from a compound of formula (1) or a protected derivative thereof, and / or said method further comprises preparing a compound of formula (3) or a protected derivative thereof from a compound of formula (2) or a protected derivative thereof, and / or said method further comprises preparing a compound of formula (4) or a protected derivative thereof from a compound of formula (3) or a protected derivative thereof, and / or said method further comprises preparing a compound of formula (5) or a protected derivative thereof from a compound of formula (4) or a protected derivative thereof, and / or said method further comprises preparing a compound of formula (6) or a protected derivative thereof from a compound of formula (5) or a protected derivative thereof, and / or said method further comprises preparing a compound of formula (7) or a protected derivative thereof from a compound of formula (6) or a protected derivative thereof, 7. The preparation method according to claim 6, wherein, said preparation of the compound of formula (2) or a protected derivative thereof is by contacting a compound of formula (1) or a protected derivative thereof with a compound of formula (1x), and / or The preparation of the compound of formula (5) or its protected derivative comprises contacting the compound of formula (4) or its protected derivative with the compound of formula (4x) or its protected derivative. and / or The preparation of the compound of formula (6) or its protected derivative comprises contacting the compound of formula (5) or its protected derivative with the compound of formula (5x). and / or The preparation of the compound or its protected derivative comprising the moiety of formula (7) comprises contacting the compound of formula (6) or its protected derivative with the compound or its protected derivative comprising the moiety of formula (6x). wherein said is selected from the group consisting of .

8. Use of the compound or its pharmaceutically acceptable salt according to any one of claims 1 to 4 in the preparation of a fibroblast activation protein inhibitor.

9. The use according to claim 8, characterized in that, The fibroblast activation protein inhibitor is a drug targeting tumors, fibrotic cells, or inflammation, particularly tumors, and more particularly the stromal fibroblasts of tumors.

10. The use according to claim 9, characterized in that, The tumor is an epithelial-derived tumor, particularly a malignant tumor, and more particularly breast cancer, gastric cancer, lung cancer, colorectal cancer, or ovarian cancer.

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