Polycyclic compound as well as intermediate, composition and application thereof

Abstract

The invention provides a polycyclic compound as well as an intermediate, a composition and application thereof. Specifically disclosed is a compound represented by formula (I) or a pharmaceutically acceptable salt thereof. The compound provided by the invention meets one or more of the following effects and advantages: (1) the structure is novel; (2) the compound has a good inhibition effect on fungi; and (3) the pharmaceutical safety is good. # imgabs0 #

Description

[0001] This application claims the priority of Chinese patent application No. 2023115662136 filed on November 22, 2023. This application cites the entire text of the above Chinese patent application. Technical Field

[0002] The present invention relates to a polycyclic compound, an intermediate thereof, a composition and use thereof. Background Art

[0003] Dihydroorotate dehydrogenase (DHODH) is an iron-containing, flavin-dependent mitochondrial enzyme that is a key enzyme in pyrimidine synthesis in nucleic acid catalysis, catalyzing the fourth step in the de novo pyrimidine biosynthesis pathway. As an enzyme associated with the electron transport chain, DHODH links mitochondrial energy, cell proliferation, ROS production, and apoptosis in certain cell types. DHODH depletion also leads to increased ROS production, decreased membrane potential, and slowed cell growth.

[0004] DHODH is a proven therapeutic target that is often used to treat autoimmune diseases, including rheumatoid arthritis and multiple sclerosis. For example, leflunomide is the first FDA-approved DHODH inhibitor. It is a synthetic isoxazole compound developed by Sanofi. In 1998, the FDA approved the marketing application of leflunomide for the treatment of rheumatoid arthritis, lupus nephritis, and psoriatic arthritis. The results of the Phase III trial of leflunomide showed that leflunomide had comparable clinical efficacy and tolerability to methotrexate and sulfasalazine, and had better efficacy and tolerability than placebo. Among 482 first-time patients, the response rate was 52%.

[0005] Teriflunomide is also a DHODH inhibitor developed by Sanofi. It was approved by the FDA in 2012. Teriflunomide inhibits the rapid differentiation of T cells, thereby slowing down the disease progression of multiple sclerosis. Based on the TEMSO study of thousands of clinical Phase III trials worldwide, the annual relapse rate of teriflunomide in the treatment of multiple sclerosis decreased by 31% compared with placebo. In addition, teriflunomide also has antiviral effects on a variety of viruses including CMV, HSV1 and BK virus. It inhibits viral replication by interfering with nucleocapsid formation and viral particle assembly.

[0006] In addition to its application in the treatment of autoimmune diseases and antiviral treatment, DHODH inhibitors can also be used in the antifungal field. Since fungi are eukaryotic organisms and have a closer relationship with humans than other microorganisms such as bacteria, it is very difficult to develop effective and non-toxic antifungal drugs. Currently, there are three main types of antifungal drugs on the market: polyenes, azoles, and echinocandins. This limits the treatment options for patients with systemic fungal infections such as invasive aspergillosis or candidiasis.

[0007] The most commonly used of the polyenes is amphotericin B. Although it radically improved patient options after its approval in the late 1950s, it is associated with severe, sometimes life-threatening side effects, including fever, heart inflammation, and kidney problems, which limits its applicability. Azoles are by far the most widely used antifungal drugs, but this class of drugs is also an inhibitor of cytochrome p450, which can cause adverse events such as liver toxicity in patients receiving other drugs. Echinocandins target glucan synthase, which means they are less toxic than the other two classes of drugs. However, echinocandins have the disadvantage of being injected, administered once a day, and each infusion takes up to an hour. Given that some patients may need months of treatment, this class of drugs is not conducive to patient compliance. Since the first echinocandin antifungal drug was approved in 2004, innovation in the antifungal field has been quite slow.

[0008] Different from the existing antifungal drug categories, Olorofim developed by F2G is the first new oral antifungal drug that exerts bactericidal activity by inhibiting the DHODH mechanism. Olorofim has an inhibitory effect on Aspergillus fumigatus DHODH (IC50: 44nM) and almost no inhibitory effect on human DHODH (>100uM). It is the only antifungal drug that has been granted breakthrough therapy designation by the FDA for multiple indications. Olorofim is currently in the open-label phase 2b clinical trial. Patients participating in the trial include patients with invasive fungal infections, aspergillosis, and refractory patients who do not respond well to existing antifungal infection drugs. Olorofim showed good tolerability during treatment, with a median treatment time of 12 weeks.

[0009] Currently, the treatment options for patients with fungal infections are very limited, and there is an urgent need for the development and market of antifungal drugs. Therefore, new antifungal drugs still need to be developed to provide more effective therapies that may save lives for patients around the world, thereby addressing the unmet medical needs caused by invasive aspergillosis and other rare fungal diseases around the world. Summary of the invention

[0010] The technical problem to be solved by the present invention is to overcome the defect of limited types of antifungal drugs in the prior art, and to this end, a polycyclic compound, an intermediate thereof, a composition and use thereof are provided. The compound of the present invention meets one or more of the following effect advantages: (1) novel structure; (2) good inhibitory effect on fungi; and (3) good pharmaceutical safety.

[0011] The present invention provides a compound represented by formula (I) or a pharmaceutically acceptable salt thereof:

[0012]

[0013] Wherein, Y is oxygen, sulfur or NR 1 ;

[0014] R 1 For hydrogen, C 1 -C 10 Alkyl or one or more R 1-1 Substituted C 1 -C 10 alkyl;

[0015] Each R 1-1 each independently is deuterium or a halogen;

[0016] R 2 is hydrogen or halogen;

[0017] X is oxygen or N-OR 3 ;

[0018] R 3 Hydrogen, -C(O)C 1 -C 10 Alkyl, C 1 -C 10 Alkyl, C 2- C 10 Alkenyl or C substituted by one or more halogen 1 -C 10 alkyl;

[0019] R 4 is a halogen;

[0020] When X is oxygen, R 1 is hydrogen or is replaced by one or more R 1-1 Substituted C 1 -C 10 alkyl.

[0021] In certain preferred embodiments of the present invention, certain groups in the compound of formula (I) or a pharmaceutically acceptable salt thereof are defined as follows, and the unmentioned groups are the same as those described in any embodiment of the present invention (referred to as "in certain embodiments of the present invention").

[0022] In some embodiments, R 1 In the C 1 -C 10 Alkyl and said one or more R 1-1 Substituted C 1 -C 10 C in the alkyl group 1 -C 10 The alkyl groups are each independently C 1 -C 6 Alkyl; the C 1 -C 6 The alkyl group is preferably a methyl group, an ethyl group, an n-propyl group, an isopropyl group, an n-butyl group, an isobutyl group or a tert-butyl group, and more preferably a methyl group.

[0023] In some embodiments, R 1-1 In the above, the halogen is fluorine, chlorine, bromine or iodine, preferably fluorine.

[0024] In some embodiments, R 2 In the above, the halogen is fluorine, chlorine, bromine or iodine, preferably fluorine.

[0025] In some embodiments, R 3 In the -C(O)C 1 -C 10 C in the alkyl group 1 -C 10 Alkyl, the C 1 -C 10 Alkyl and the C 1 -C 10 C in the alkyl group 1 -C 10 The alkyl groups are each independently C 1 -C 6 Alkyl; the C 1 -C 6 The alkyl group is preferably a methyl group, an ethyl group, an n-propyl group, an isopropyl group, an n-butyl group, an isobutyl group or a tert-butyl group, and more preferably a methyl group, an ethyl group, an isopropyl group or a tert-butyl group.

[0026] In some embodiments, R 3 In the C 2- C 10 Alkenyl is C 2- C 6 Alkenyl, the C 2- C 6 The alkenyl group is preferably a vinyl group, More preferably

[0027] In some embodiments, R 3In the case of C substituted by one or more halogens 1 -C 10 The halogen in the alkyl group is fluorine, chlorine, bromine or iodine, preferably fluorine.

[0028] In some embodiments, R 4 In the above, the halogen is fluorine, chlorine, bromine or iodine, preferably fluorine.

[0029] In some embodiments, the compound of formula (I) is a compound of formula (IA) or (IB):

[0030]

[0031] Y, R 2 and R 3 The definitions are as mentioned above.

[0032] In some embodiments, in Formula (IA), Y is oxygen, sulfur or NR 1 ;

[0033] R 1 For hydrogen, C 1 -C 6 Alkyl or one or more R 1-1 Substituted C 1 -C 6 alkyl;

[0034] Each R 1-1 each independently is deuterium or a halogen;

[0035] R 2 is hydrogen or halogen;

[0036] R 3 Hydrogen, -C(O)C 1 -C 6 Alkyl, C 1 -C 6 Alkyl or C 2- C 6 Alkenyl.

[0037] In some embodiments, in Formula (IB), Y is oxygen, sulfur or NR 1 ;

[0038] R 1 is hydrogen or is replaced by one or more R 1-1 Substituted C 1 -C 6 alkyl;

[0039] Each R 1-1 Each independently is deuterium;

[0040] R 2 is hydrogen or halogen.

[0041] In some embodiments, in formula (I-A), Y is oxygen, sulfur or N-CH 3 .

[0042] In some embodiments, in formula (I-A), R 2 is hydrogen or -F.

[0043] In some embodiments, in formula (I-A), R 3 is hydrogen, -CH 3 、-CH 2 CH 3 、

[0044] In some embodiments, in formula (I-B), Y is oxygen, sulfur or N-CD 3 .

[0045] In some embodiments, in formula (I-B), R 2 is hydrogen or -F.

[0046] In some embodiments, the compound represented by formula (I) is any of the following compounds:

[0047]

[0048]

[0049] The present invention also provides a compound represented by formula (II);

[0050]

[0051] wherein the definitions of X and Y are as described above.

[0052] In some embodiments, the compound represented by formula (II) is any of the following compounds:

[0053]

[0054] The present invention also provides a pharmaceutical composition, which comprises: the above-mentioned compound represented by formula (I) or a pharmaceutically acceptable salt thereof (effective therapeutic amount), and a pharmaceutically acceptable excipient.

[0055] The present invention also provides the use of a substance A in the preparation of a drug for preventing and / or treating a disease caused by fungal infection; the substance A is the above-mentioned compound represented by formula (I) or a pharmaceutically acceptable salt thereof (effective therapeutic amount), or the above-mentioned pharmaceutical composition.

[0056] In some embodiments, the fungus is preferably Aspergillus and / or Candida albicans (C.albicans), and the Aspergillus is preferably Aspergillus flavus (A.flavus), Aspergillus fumigatus (A.fumigatus), Aspergillus niger (A.niger) or Aspergillus terreus (A.terreus) and the like.

[0057] Terminology

[0058] In the present invention, the term "pharmaceutically acceptable salt" refers to a salt obtained by reacting a compound with a pharmaceutically acceptable acid or base. When the compound contains a relatively acidic functional group, a base addition salt can be obtained by contacting the compound with a sufficient amount of a pharmaceutically acceptable base in a suitable inert solvent. When the compound contains a relatively basic functional group, an acid addition salt can be obtained by contacting the compound with a sufficient amount of a pharmaceutically acceptable acid in a suitable inert solvent. For details, please refer to Handbook of Pharmaceutical Salts: Properties, Selection, and Use (P. Heinrich Stahl, Camille G. Wermuth, 2011, 2nd Revised Edition).

[0059] In the present invention, the structural fragment It means that the structural fragment is connected to the rest of the molecule through this bond. For example, It refers to isopropyl.

[0060] In the present invention, the "-" at the end of a group means that the group is connected to the rest of the molecule through this site. For example, -OH refers to a hydroxyl group.

[0061] In the present invention, the term "one or more" refers to 1, 2, 3, 4 or 5, such as 1, 2 or 3.

[0062] In the present invention, the term "halogen" refers to fluorine, chlorine, bromine or iodine.

[0063] In the present invention, the term "alkyl" refers to a group having a specified number of carbon atoms (e.g., C 1 -C 10 , C 1 -C 6 ) is a linear or branched, saturated, monovalent hydrocarbon group. The alkyl group includes, but is not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, n-hexyl, etc.

[0064] In the present invention, the term "alkenyl" refers to a group having a specified number of carbon atoms (e.g., C 2- C 10 , C 2~C 6 ) is a linear or branched, unsaturated, monovalent hydrocarbon group having one or more (e.g., 1, 2, or 3) carbon-carbon sp 2 Double bond. Alkenyl groups include but are not limited to: vinyl, wait.

[0065] In the present invention, the term "pharmaceutically acceptable excipients" refers to all substances contained in pharmaceutical preparations other than active pharmaceutical ingredients, which are generally divided into two categories: excipients and additives. For details, please refer to the Pharmacopoeia of the People's Republic of China (2020 Edition) and Handbook of Pharmaceutical Excipients (Paul J Sheskey, Bruno C Hancock, Gary P Moss, David J Goldfarb, 2020, 9th Edition).

[0066] Without violating the common sense in the art, the above-mentioned preferred conditions can be arbitrarily combined to obtain the preferred embodiments of the present invention.

[0067] The reagents and raw materials used in the present invention are commercially available.

[0068] The positive and progressive effects of the present invention are that the compounds of the present invention satisfy one or more of the following advantages: novel structure, good inhibitory effect on fungi, and good pharmaceutical safety. DETAILED DESCRIPTION

[0069] The present invention is further described below by way of examples, but the present invention is not limited to the scope of the examples. The experimental methods in the following examples without specifying specific conditions are carried out according to conventional methods and conditions, or selected according to the product specifications.

[0070] Example 1 Synthesis of methyl 2-(1,5-dimethyl-3-phenyl-1H-pyrrol-2-yl)-2-oxoacetate (Intermediate IIa)

[0071]

[0072] Step 1: 4-Bromo-1-methyl-1H-pyrrole-2-carbaldehyde (IIa-1)

[0073] 4-Bromo-1H-pyrrole-2-carboxaldehyde (10 g, 57.47 mmol) was added to N,N-dimethylformamide (150 mL), cooled to 0°C, 60% sodium hydride (4.6 g, 114.94 mmol) was added and reacted for 1 h, then iodomethane (12.2 g, 86.21 mmol) was added and reacted for 3 h, then the reaction solution was quenched with saturated sodium carbonate solution (200 mL), and then 200 mL of water was added to dilute the reaction solution, and extracted with ethyl acetate (100 mL x 4) to obtain an organic phase, which was washed with saturated brine (100 mL x 2), dried, spin-dried, mixed, and purified by silica gel column chromatography to obtain compound IIa-1 (8.78 g, yield 81.3%). LC-MS (ESI, m / z) 188.0 [M+H] + ; 1 H NMR (400 MHz, DMSO-d 6 )δ9.49(d,J=1.2Hz,1H),7.40-7.41(m,1H),7.09(d,J=2.0Hz,1H),3.87(s,3H).

[0074] Step 2: 1-Methyl-4-phenyl-1H-pyrrole-2-carbaldehyde (IIa-2)

[0075] Under nitrogen protection, compound IIa-1 (15 g, 79.78 mmol) and phenylboronic acid (11.7 g, 95.73 mmol) were dissolved in a mixed solvent of 1,4-dioxane and water (350 mL, v / v = 10:1), cesium carbonate (57.2 g, 175.51 mmol) and 1,1-bis(diphenylphosphino)ferrocenepalladium dichloride (3.5 g, 4.79 mmol) were added, and the temperature was raised to 95°C and stirred for 5 hours. The reaction was monitored by TLC. The reaction solution was filtered through diatomaceous earth, and the filter cake was washed with ethyl acetate. The filtrate was concentrated, the sample was mixed, and the compound IIa-2 (10.38 g, yield 70.3%) was obtained by purification and separation by silica gel column chromatography. LC-MS (ESI, m / z) 186.1 [M+H] + ; 1 H NMR (400 MHz, DMSO-d 6 )δ9.58(d,J=0.8Hz,1H),7.74(s,1H),7.59-7.61(m,2H),7.40(d,J=2.0Hz,1H),7.35-7.38(m,2H),7.18-7.23(m,1H),3.92(s,3H).

[0076] Step 3: 1,2-Dimethyl-4-phenyl-1H-pyrrole (IIa-3)

[0077] Compound IIa-2 (10.0 g, 53.99 mmol), potassium hydroxide (9.1 g, 161.96 mmol), and hydrazine hydrate (9.5 g, 161.96 mmol, 85% (wt)) were dissolved in diethylene glycol (120 mL), heated to 130 ° C and stirred for 3 h, cooled to room temperature, diluted with water (200 mL), and extracted with ethyl acetate (200 mL*4). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated, and purified by silica gel column chromatography (PE / EA=20:1) to obtain compound IIa-3 (6.7 g, yield 72.5%), LC-MS (ESI, m / z) 172.1 [M+H] + ; 1 H NMR (300 MHz, DMSO-d 6 )δ7.44-7.47(m,2H),7.29(t,J=7.5Hz,2H),7.06-7.11(m,2H),6.18(d,J=0.6Hz,1H),3.54(s,3H),2.21(s,3H).

[0078] Step 4: Methyl 2-(1,5-dimethyl-3-phenyl-1H-pyrrol-2-yl)-2-oxoacetate (Intermediate IIa)

[0079] Dissolve oxalyl chloride (16.7 g, 116.79 mmol) in dichloromethane (100 mL) and keep it at -10 ° C. Then dissolve the intermediate IIa-3 (20 g, 116.79 mmol) in dichloromethane (200 mL) and slowly add it to the above solution. Keep the reaction liquid temperature at -10 ° C to 0 ° C. After stirring at 0 ° C for 1 hour, slowly add the reaction liquid dropwise to the sodium bicarbonate aqueous solution cooled in an ice bath, continue stirring for 30 minutes, and separate the liquids. The aqueous phase is extracted twice with dichloromethane (250 mL), and then the pH value is adjusted to 2-3 with 20% sulfuric acid. Solids precipitate, stir in an ice bath for 30 minutes, filter, and wash with water. The filter cake is dried under vacuum to obtain the target compound IIa (17.83 g, yield 62.8%). LC-MS (ESI, m / z) 244.1 [M+H] + .

[0080] Example 2 Synthesis of 2-(5-methyl-1-(methyl-d3)-3-phenyl-1H-pyrrol-2-yl)-2-oxoacetic acid (IIb)

[0081]

[0082] Step 1: 4-Bromo-1-methyl-d3-1H-pyrrole-2-carbaldehyde (IIb-1)

[0083] Referring to step 1 of Example 1, 4-bromo-1H-pyrrole-2-carboxaldehyde (3.5 g, 20.1 mmol) was subjected to alkylation reaction with deuterated iodomethane (3.21 g, 22.1 mmol) to obtain compound IIb-1 (3.2 g, white solid, yield 83.7%). LC-MS (ESI, m / z): 191.0 / 192.0 [M+H] + ; 1 H NMR (300 MHz, DMSO-d 6 )δ9.55(d,J=0.6Hz,1H),7.46(d,J=0.9Hz,1H),7.15(d,J=1.8Hz,1H).

[0084] Step 2: 1-(Methyl-d3)-4-phenyl-1H-pyrrole-2-carbaldehyde (IIb-2)

[0085] Referring to step 2 of the example, IIa-1 (1.0 g, 90%, 4.8 mmol) and phenylboronic acid (0.88 g, 7.2 mmol) were subjected to Suzuki coupling reaction to obtain compound IIb-2 (0.48 g, yellow solid, yield 62.8%). LC-MS (ESI, m / z): 189.1 [M+H] + ; 1 H NMR (400 MHz, DMSO-d 6 )δ9.58(s,1H),7.74(dd,J=2.0,0.8Hz,1H),7.58-7.61(m,2H),7.41(d,J=2.0Hz,1H),7.34-7.38(m,2H),7.18-7.22(m,1H).

[0086] Step 3: 2-Methyl-1-(methyl-d3)-4-phenyl-1H-pyrrole (IIb-3)

[0087] Referring to the operation of step 3 of Example 1, compound IIb-2 (480 mg, 2.55 mmol) and hydrazine hydrate (637 mg, 7.65 mmol) were subjected to Wolf-Kisler reduction reaction to obtain compound IIb-4 (200 mg, yellow solid, yield 42.8%). LC-MS (ESI, m / z) 175.1 [M+H] + ; 1 H NMR (400 MHz, DMSO-d 6 )δ7.39-7.41(m,2H),7.22-7.27(m,2H),7.02-7.06(m,1H),6.96-6.97(m,1H),6.11-6.12(m,1H),2.20(s,3H).

[0088] Step 4 2-(5-methyl-1-(methyl-d3)-3-phenyl-1H-pyrrol-2-yl)-2-oxoacetic acid (IIb)

[0089] The synthetic operation was obtained by referring to step 4 of Example 1, and compound IIb-4 was subjected to Friedel-Crafts acylation reaction with oxalyl chloride to obtain compound IIb. LC-MS (ESI, m / z) 247.1 [M+H ]+ .

[0090] Example 3 Synthesis of 2-(1,5-dimethyl-3-phenyl-1H-pyrrol-2-yl)-2-(hydroxyimino)acetic acid (IIc)

[0091]

[0092] Step 1: Methyl 2-(1,5-dimethyl-3-phenyl-1H-pyrrol-2-yl)-2-oxoacetate (IIc-1)

[0093] Compound IIa-3 (5.0 g, 29.20 mmol) was dissolved in dichloromethane (100 mL) and cooled to 0°C. Under nitrogen protection, methyl 2-chloro-2-oxoacetate (3.9 g, 32.12 mmol) was slowly added, and then the temperature was slowly raised to room temperature and the reaction was allowed to proceed overnight. TLC monitored the completion of the reaction, and the reaction mixture was added to a sodium bicarbonate solution, extracted with dichloromethane, and the dichloromethane phase was concentrated to dryness and purified by silica gel column chromatography to obtain compound IIc-1 (6.48 g, yield 86.3%). LC-MS (ESI, m / z) 258.1 [M+H] + .

[0094] Step 2: 2-(1,5-dimethyl-3-phenyl-1H-pyrrol-2-yl)-2-(hydroxyimino)acetic acid methyl ester (IIc-2)

[0095] In a 50mL round-bottom flask, compound IIc-1 (3.0g, 11.66mmol), methoxyamine hydrochloride (2.14g, 25.65mmol), potassium acetate (5.1g, 51.30mmol), ethanol (30mL) and water (5mL) were added. The reaction solution was heated to 85°C and reacted for 3.5 hours. After the reaction was completed, ethanol was removed by vacuum distillation, and the remaining aqueous solution was extracted with dichloromethane. The organic phase was dried with sodium sulfate, filtered and concentrated, and purified by silica gel column chromatography to obtain compound IIc-2 (1.89g, yield 59.5%). LC-MS (ESI, m / z) 287.1 [M+H] + .

[0096] Step 3: 2-(1,5-dimethyl-3-phenyl-1H-pyrrol-2-yl)-2-(hydroxyimino)acetic acid (IIc)

[0097] Compound IIc-2 (1.5 g, 5.51 mmol), lithium hydroxide (396 mg, 16.53 mmol), methanol (15 mL) and water (3 mL) were added to a 100 mL three-necked flask. After dissolution, the system was reacted at room temperature for 3 hours and the reaction was monitored by TLC. After the reaction was completed, the methanol was removed by concentration under reduced pressure, and the pH was adjusted to 3-4 with dilute hydrochloric acid, extracted with ethyl acetate, and the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the target compound IIc (1.1 g, yield 77.3%). LC-MS (ESI, m / z) 273.1 [M+H] + .

[0098] Example 4 Synthesis of 2-(5-methyl-3-phenylthiophen-2-yl)-2-oxoacetic acid (IId)

[0099]

[0100] Step 1: 2-Methyl-4-phenylthiophene (IId-1)

[0101] Under nitrogen protection, 4-bromo-2-methylthiophene (15 g, 84.75 mmol) and phenylboronic acid (11.7 g, 95.73 mmol) were dissolved in a mixed solvent of 1,4-dioxane / water (350 mL, v / v = 10:1), and then cesium carbonate (57.2 g, 175.51 mmol) and 1,1-bis(diphenylphosphino)ferrocenepalladium dichloride (3.5 g, 4.79 mmol) were added, and the temperature was raised to 95°C for 5 hours. The reaction was monitored by TLC. The reaction solution was filtered through diatomaceous earth, and the filter cake was washed with ethyl acetate. The filtrate was dried, the sample was mixed, and the compound IId-1 (10.41 g, yield 70.2%) was isolated by column purification (PE / EA = 10 / 1). LC-MS (ESI, m / z) 175.1 [M+H] + ; 1 H NMR (400 MHz, CDCl 3 )δ7.52-7.45(m,2H),7.33-7.26(m,2H),7.23-7.16(m,1H),7.12(d,J=1.5Hz,1H),6.98(p,J=1.2Hz,1H),2.45(d,J=1.1Hz,3H).

[0102] Step 2: Synthesis of methyl 2-(5-methyl-3-phenylthiophen-2-yl)-2-oxyacetate (IId-2)

[0103] Compound IId-1 (5.0 g, 28.73 mmol) was dissolved in dichloromethane (100 mL), cooled to 0 ° C, and methyl 2-chloro-2-oxoacetate (4.86 g, 39.63 mmol) was slowly added under nitrogen protection, and then the temperature was slowly raised to room temperature and reacted overnight. TLC monitored the end of the reaction, the reaction mixture was added to a sodium bicarbonate solution, extracted with dichloromethane, the dichloromethane phase was concentrated to dryness, and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 20: 1 to 1: 1) to obtain compound IIc-2 (5.27 g, yield 70.5%), LCMS (m / z): 261.1 [M+H] + ; 1 HNMR (400 MHz, DMSO-d 6 )δ7.46-7.44(m,3H),7.36-7.33(m,2H),7.11(d,J=0.8Hz,1H),2.25(d,J=0.8Hz,3H).

[0104] Step 3: Synthesis of 2-(5-methyl-3-phenylthiophen-2-yl)-2-oxoacetic acid (IId)

[0105] Compound IId-2 (2.0 g, 7.69 mmol), lithium hydroxide (348 mg, 14.53 mmol), methanol (15 mL) and water (3 mL) were added to a 50 mL three-necked flask. After dissolution, the system was reacted at room temperature for 3 hours, and the reaction was monitored by TLC. After the reaction was completed, the methanol was removed by concentration under reduced pressure, and the pH was adjusted to 3-4 with dilute hydrochloric acid, extracted with ethyl acetate, and the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the target compound IId (1.1 g, yield 58.2%), LCMS (m / z): 247.0 [M+H] + ; 1 HNMR (400 MHz, DMSO-d 6 )δ14.12(s,1H),7.42 -7.38(m,5H),2.56(s,3H).

[0106] Example 5 Synthesis of 2-(5-methyl-3-phenylfuran-2-yl)-2-oxoacetic acid (IIe)

[0107]

[0108] Step 1: 4-phenylfuran-2-carbaldehyde (IIe-1)

[0109] The experimental operation was the same as step 2 of Example 1. 4-bromofuran-2-carboxaldehyde (15 g, 79.78 mmol) and phenylboronic acid (11.7 g, 95.73 mmol) were subjected to Suzuki coupling reaction to obtain compound IIe-1 (10.38 g, yield 70.3%). LC-MS (ESI, m / z) 173.1 [M+H] + .

[0110] Step 2: 2-Methyl-4-phenylfuran (IIe-2)

[0111] The experimental operation was the same as step 3 of Example 1. The aldehyde group of compound IIe-1 (10.0 g, 53.99 mmol) was reduced with hydrazine hydrate to obtain compound IIe-2 (6.7 g, yield 72.5%). LC-MS (ESI, m / z) 159.1 [M+H] + . 1 H NMR (300MHz, Chloroform-d) δ7.64(d,J=1.0Hz,1H),7.56-7.47(m,2H),7.47-7.36(m,2H),7.36–7.25(m,1H),6.37(q,J=1.1Hz,1H),2.40(d,J=1.1Hz,3H).

[0112] Step 3: Methyl 2-(5-methyl-3-phenylfuran-2-yl)-2-oxoacetate (IIe-3)

[0113] Referring to the operation of step 2 of Example 4, compound IIe-2 was subjected to Friedel-Crafts acylation reaction with methyl 2-chloro-2-oxoacetate to obtain compound IIe-3. LC-MS (ESI, m / z) 245.1 [M+H] + ; 1 H-NMR (300MHz, CDCl 3 )δ7.46-7.48(m,2H),7.33-7.37(m,3H),6.32(d,J=0.6Hz,1H),3.55(s,3H),2.39(s,3H).

[0114] Step 4: 2-(5-methyl-3-phenylfuran-2-yl)-2-oxoacetic acid (IIe)

[0115] Referring to step 2 of Example 4, compound IIe-2 was hydrolyzed with lithium hydroxide to obtain IIe (17.83 g, yield 62.8%). LC-MS (ESI, m / z) 231.1 [M+H] + ; 1 H-NMR (300 MHz, DMSO-d 6)δ7.73-7.75(m,2H),7.38-7.44(m,3H),6.68(d,J=0.6Hz,1H),2.37(s,3H).

[0116] Example 6 Synthesis of 4-(4-(5-fluoropyrimidin-2-yl)piperazin-1-yl)aniline (IIIa)

[0117]

[0118] Step 1: tert-Butyl 4-(5-fluoropyrimidin-2-yl)piperazine-1-carboxylate (IIIa-1)

[0119] 2-Chloro-5-fluoropyrimidine (10.0 g, 75.17 mmol) and tert-butyl piperazine-1-carboxylate (10.0 g, 53.69 mmol) were dissolved in ethanol (120 mL), diisopropylethylamine (41.6 g, 322.14 mmol) was added, and the temperature was raised to 80°C for 16 hours. The reaction solution was filtered, and the filtrate was concentrated to dryness and purified by silica gel column chromatography to obtain compound IIIa-1 (10.76 g, yield 71.0%). LC-MS (ESI, m / z) 283.1 [M+H] + . 1 H NMR (300 MHz, CDCl 3 )δ8.25(s,2H),3.79(d,J=5.1Hz,4H),3.54(d,J=5.4Hz,1H),1.53(s,9H).

[0120] Step 2: 5-Fluoro-2-(piperazin-1-yl)pyrimidine hydrochloride (IIIa-2)

[0121] Compound IIIa-1 (10.0 g, 25.2 mmol), a dioxane solution of hydrogen chloride (4 M, 90 mL) and dichloromethane (20 mL) were added to a 250 mL single-mouth bottle, and then reacted at room temperature for 2 hours and monitored by TLC. After the reaction was completed, the reaction solution was concentrated to dryness, neutralized with sodium bicarbonate solution, extracted with dichloromethane, and then concentrated to obtain compound IIIa-2 (5.95 g, yield 92.2%). LC-MS (ESI, m / z) 183.2 [M+H] + .

[0122] Step 3: 5-Fluoro-2-(4-(4-nitrophenyl)piperazin-1-yl)pyrimidine (IIIa-3)

[0123] Compound IIIa-2 (10.0 g, 54.88 mmol) and 1-fluoro-4-nitrobenzene (7.7 g, 54.88 mmol) were dissolved in ethylene glycol dimethyl ether (100 mL), potassium carbonate (15.2 g, 109.77 mmol) was added, and the temperature was raised to 100 ° C. and stirred for 16 hours. The reaction solution was added to water, and the solid matter was precipitated and filtered. The obtained filter cake was centrifuged and dried to obtain compound IIIa-3 (12.7 g, yield 76.3%). LC-MS (ESI, m / z) 304.1 [M+H] + ; 1 H NMR (400 MHz, DMSO-d 6 )δ8.50(d,J=0.8Hz,2H),8.07(d,J=9.2Hz,2H),7.04(d,J=9.6Hz,2H),3.83-3.86(m,4H),3.59-3.62(m,4H).

[0124] Step 4: 4-(4-(5-fluoropyrimidin-2-yl)piperazin-1-yl)aniline (IIIa)

[0125] Compound IIIa-3 (10.0 g, 32.97 mmol) was added to a reaction flask and dissolved in isopropanol (100 mL). Then 10% palladium on carbon (2.1 g, 1.98 mmol) was added. The reaction system was replaced with hydrogen. The reaction solution was stirred at room temperature overnight under a hydrogen (15 Psi) atmosphere. TLC monitoring showed that the reaction was complete. The reaction filtrate was concentrated under reduced pressure by suction filtration to obtain a crude product, which was purified by silica gel column chromatography to obtain a compound (8.21 g, yield 91.0%). LC-MS (ESI, m / z) 274.1 [M+H ]+ ; 1 H NMR (300 MHz, DMSO-d 6 )δ8.48(s,2H),6.76(d,J=8.7Hz,2H),6.54(d,J=8.7Hz,2H),4.63(s,2H),3.80-3.84(m,4H),2.96-2.99(m,4H).

[0126] Example 7 Synthesis of 3-fluoro-4-(4-(5-fluoropyrimidin-2-yl)piperazin-1-yl)aniline (IIIb)

[0127]

[0128] Step 1: 5-Fluoro-2-(4-(2-fluoro-4-nitrophenyl)piperazin-1-yl)pyrimidine (IIIb-1)

[0129] Compound IIIa-3 (10.0 g, 54.88 mmol) was dissolved in tetrahydrofuran (100 mL), diisopropylethylamine (39.4 mL, 219.54 mmol) and 3,4-difluoronitrobenzene (8.7 g, 54.88 mmol) were added, and the temperature was raised to 60 ° C and stirred for 12 hours. The reaction solution was poured into water and extracted with ethyl acetate (150 mL x 3 times). The extract was washed three times with saturated sodium chloride solution (150 mL x 3 times), dried with anhydrous magnesium sulfate, filtered and concentrated to obtain an orange-yellow solid. Recrystallization was performed with acetone: water (v / v = 9:1) to obtain the target compound (15.78 g, yield 89.5%). LC-MS (ESI, m / z) 322.1 [M+H] + ; 1 HNMR (400 MHz, DMSO-d 6 )δ8.50(d,J=0.8Hz,2H),8.01-8.06(m,2H),7.19-7.23(m,1H),3.85-3.87(m,4H),3.38-3.41(m,4H).

[0130] Step 2: 3-Fluoro-4-(4-(5-fluoropyrimidin-2-yl)piperazin-1-yl)aniline (IIIb)

[0131] Compound IIIb-1 (10.0 g, 31.12 mmol) was dissolved in isopropanol (100 mL), and then 10% palladium carbon (2.0 g, 1.87 mmol) was added. The reaction system was replaced with hydrogen, and the reaction solution was stirred at room temperature overnight under a hydrogen (1 atm) atmosphere. TLC monitoring showed that the reaction was complete. The reaction solution was filtered and the filtrate was concentrated to obtain compound IIIb (8.11 g, yield 89.5%). LC-MS (ESI, m / z) 292.1 [M+H] + ; 1 H NMR (400 MHz, DMSO-d 6 )δ8.46(s,2H),6.75 -6.81(m,1H),5.00(s,2H),6.29-6.38(m,2H),3.85-3.87(m,4H),2.86 -2.90(m,4H).

[0132] Example 8 Synthesis of 2-(1,5-dimethyl-3-phenyl-1H-pyrrol-2-yl)-N-(4-(4-(5-fluoropyrimidin-2-yl)piperazin-1-yl)phenyl)-2-oxoacetamide (I-1, comparative compound Olorofim)

[0133]

[0134] Compound IIa (2.0 g, 7.32 mmol) was dissolved in N,N-dimethylformamide (20 mL), HATU (3.3 g, 8.78 mmol) and diisopropylethylamine (2.8 g; 21.95 mmol) were added, and the mixture was stirred at room temperature for 10 min, and then compound IIIa (1.78 g, 7.32 mmol) was added and stirred at room temperature for 2 hours. Water (80 mL) was added to quench the reaction, and the aqueous phase was taken with dichloromethane (50 mL*3), the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated, and the residue was purified by silica gel column chromatography (dichloromethane: ethyl acetate = 50:1 to 1:1) to obtain compound I-1 (2.85 g, yield 78.1%). LC-MS (ESI, m / z) 499.2 [M+H] + ; 1 HNMR (400 MHz, DMSO-d 6 )δ10.21(s,1H),8.48(d,J=0.4Hz,2H),7.22-7.25(m,2H),7.09-7.14(m,3H),7.03-7.06( m,2H),6.80-6.83(m,2H),6.10(s,2H),3.79-3.82(m,7H),3.11-3.14(m,4H),2.32(s,3H).

[0135] Example 9 Synthesis of N-(4-(4-(5-fluoropyrimidin-2-yl)piperazin-1-yl)phenyl)-2-(5-methyl-1-(methyl-d3)-3-phenyl-1H-pyrrol-2-yl)-2-oxoacetamide (I-2)

[0136]

[0137] Referring to the synthesis operation of Example 8, compound IId (2.0 g, 7.66 mmol) was condensed with compound IIIa (2.1 g, 7.66 mmol) to obtain compound I-2 (2.85 g, yield 78.1%). LC-MS (ESI, m / z) 502.2 [M+H] + ; 1 H NMR (400 MHz, DMSO-d 6 )δ10.21(s,1H),8.48(s,2H),7.24(dd,J=7.6,1.6Hz,2H),7.09-7.14(m,3H),7.05(d,J=8.8H z,2H),6.82(d,J=9.2Hz,2H),6.10(s,1H),3.79-3.82(m,4H),3.11-3.14(m,4H),2.32(s,3H).

[0138] Example 10 Synthesis of N-(4-(4-(5-fluoropyrimidin-2-yl)piperazin-1-yl)phenyl)-2-(5-methyl-3-phenylthiophen-2-yl)-2-oxoacetamide (I-3)

[0139]

[0140] Referring to the synthesis operation of Example 8, compound IId (2.0 g, 7.66 mmol) was condensed with compound IIIa (2.1 g, 7.66 mmol) to obtain compound I-3 (2.85 g, yield 78.1%). LC-MS (ESI, m / z) 502.2 [M+H] + ; 1 HNMR (400 MHz, DMSO-d 6 )δ10.52(s,1H),8.49(d,J=0.4Hz,2H),7.47-7.45(m,2H),7.43-7.41(m,2H),7.35-7.31(m,3H ),7.04(d,J=0.8Hz,1H),6.95-6.93(m,2H),3.84-3.81(m,4H),3.19-3.16(m,4H),2.57(s,2H).

[0141] Example 11 Synthesis of N-(3-fluoro-4-(4-(5-fluoropyrimidin-2-yl)piperazin-1-yl)phenyl)-2-(5-methyl-3-phenylthiophen-2-yl)-2-oxoacetamide (I-4)

[0142]

[0143] Referring to the synthesis operation of Example 8, compound IId (2.0 g, 7.66 mmol) was condensed with compound IIIb (2.1 g, 7.66 mmol) to obtain compound I-4 (2.85 g, yield 78.1%). LC-MS (ESI, m / z) 520.2 [M+H] + .

[0144] Example 12 Synthesis of N-(4-(4-(5-fluoropyrimidin-2-yl)piperazin-1-yl)phenyl)-2-(5-methyl-3-phenylfuran-2-yl)-2-oxoacetamide (I-5)

[0145]

[0146] Referring to the synthesis operation of Example 8, compound IIe (2.0 g, 7.66 mmol) was condensed with compound IIIa (2.1 g, 7.66 mmol) to obtain compound I-5 (2.85 g, yield 78.1%). LC-MS (ESI, m / z) 486.2 [M+H] + ; 1 H-NMR (400MHz, CDCl 3 )δ8.65(s,1H),8.22(s,2H),7.52-7.57(m,3H),7.39-7.42(m,2H),6.97(brs,2H),6.40(s,1H),3.94(br s,4H),3.23-3.24(m,4H),2.50(s,3H).

[0147] Example 13 Synthesis of 2-(1,5-dimethyl-3-phenyl-1H-pyrrol-2-yl)-N-(4-(4-(5-fluoropyrimidin-2-yl)piperazin-1-yl)phenyl)-2-(hydroxyimino)acetamide (I-6)

[0148]

[0149] Compound I-1 (1.0 g, 2.01 mmol) was dissolved in a mixed solvent of ethanol (15 mL) and water (3 mL), hydroxylamine hydrochloric acid (306 mg, 4.41 mmol) and potassium acetate (866 mg, 8.83 mmol) were added, and the temperature was heated to 85 ° C and stirred for 3.5 hours. After the reaction was completed, ethanol was removed by distillation under reduced pressure, and the remaining aqueous solution was extracted with dichloromethane. The organic phase was dried with sodium sulfate, filtered and concentrated, and purified by silica gel column chromatography (dichloromethane: ethyl acetate = 50: 1 to 1: 1) to obtain compound I-6 (788 mg, yield 76.5%). LC-MS (ESI, m / z) 514.2 [M+H] + ; 1 H NMR (400 MHz, DMSO-d 6 )δ12.21(s,1H),9.98(s,1H),8.47(d,J=0.8Hz,2H),7.48(d,J=9.2Hz,2H),7.29(dd,J=8.4,1.2Hz,2H),7.20-7.24(m,2H),7. 04-7.08(m,1H),6.91(d,J=9.2Hz,2H),6.14(d,J=0.8Hz,1H),3.80-3.82(m,4H),3.31(s,3H),3.12-3.15(m,4H),2.25(s,3H).

[0150] Example 14 Synthesis of 2-(acetoxyimino)-2-(1,5-dimethyl-3-phenyl-1H-pyrrol-2-yl)-N-(4-(4-(5-fluoropyrimidin-2-yl)piperazin-1-yl)phenyl)acetamide (I-7)

[0151]

[0152] Compound I-6 (1.0 g, 1.95 mmol), pyridine (462 mg; 5.84 mmol) and N, N-dimethylpyridin-4-amine (48 mg; 0.389 mmol) were dissolved in dichloromethane (12 mL), stirred at room temperature for 10 min, and then acetic anhydride (219 mg, 2.14 mmol) was added and stirred at room temperature for 2 hours. Water (80 mL) was added to quench the reaction, and the aqueous phase was taken with dichloromethane (50 mL*3), the organic phases were combined, washed with saturated brine, dried with anhydrous sodium sulfate, and purified by silica gel column chromatography (dichloromethane: ethyl acetate = 50: 1 to 1: 1) to obtain compound I-7 (789 mg, yield 72.9%). LC-MS (ESI, m / z) 556.2 [M+H] + ; 1 HNMR (400 MHz, DMSO-d 6 )δ10.62(s,1H),8.48 -8.49(m,2H),7.49-7.52(m,2H),7.26-7.27(m,4H),7.15-7.19(m,1H),6.96-6.98(m,1H) ,6.18(s,2H),3.81-3.83(m,4H),3.37(s,3H),3.17-3.19(m,4H),2.26(s,3H),1.78(s,3H)

[0153] Example 15 Synthesis of 2-(1,5-dimethyl-3-phenyl-1H-pyrrol-2-yl)-N-(4-(4-(5-fluoropyrimidin-2-yl)piperazin-1-yl)phenyl)-2-(methoxyimino)acetamide (I-8)

[0154]

[0155] Compound I-1 (1.0 g, 2.01 mmol) and O-methylhydroxylamine hydrochloride (368 mg, 4.41 mmol) were dissolved in a mixed solvent of ethanol (15 mL) and water (3 mL), potassium acetate (866 mg, 8.83 mmol) was added, and the temperature was raised to 85 ° C and stirred for 3.5 hours. After the reaction was completed, ethanol was removed by distillation under reduced pressure, and the remaining aqueous solution was extracted with dichloromethane, the organic phase was dried with sodium sulfate, filtered and concentrated, and purified by silica gel column chromatography (dichloromethane: ethyl acetate = 50: 1 to 1: 1) to obtain compound I-8 (815 mg, yield 77.1%). LC-MS (ESI, m / z) 528.2 [M+H] + ; 1 H NMR (400 MHz, DMSO-d 6 )δ10.11(s,0.5H),10.07(s,0.5H),8.48(t,J=1.1Hz,2H),7.47-7.40(m,1H),7. 39-7.32(m,1H),7.31-7.14(m,3H),7.13-7.03(m,1H),6.95-6.85(m,2H),6.13( d,J=1.0Hz,0.5H),6.01(d,J=0.9Hz,0.5H),3.96(s,1H),3.86(s,1.5H),3.81(t ,J=5.2Hz,4H),3.57(s,1.5H),3.32(s,3H),3.14(q,J=4.8Hz,4H),2.24(s,3H).

[0156] Example 16 Synthesis of 2-(1,5-dimethyl-3-phenyl-1H-pyrrol-2-yl)-N-(4-(4-(5-fluoropyrimidin-2-yl)piperazin-1-yl)phenyl)-2-(methoxyimino)acetamide (I-8)

[0157]

[0158] Compound IIc (2.g, 7.74mmol), HATU (3.5g, 9.29mmol) and diisopropylethylamine (3.0g; 23.23mmol) were dissolved in N,N-dimethylformamide (20mL), stirred at room temperature for 10min, and then compound IIIa (2.1g, 7.74mmol) was added and stirred at room temperature for 2 hours. Water (80mL) was added to quench the reaction, and the aqueous phase was taken with dichloromethane (50mL*3), the organic phases were combined, washed with saturated brine, dried with anhydrous sodium sulfate, and purified by silica gel column chromatography (dichloromethane: ethyl acetate = 50:1 to 1:1) to obtain compound I-8 (2.72g, yield 68.4%). LC-MS (ESI, m / z) 528.2 [M+H] + ; 1 HNMR (400 MHz, DMSO-d 6 )δ10.11(s,0.5H),10.07(s,0.5H),8.48(t,J=1.1Hz,2H),7.47-7.40(m,1H),7. 39-7.32(m,1H),7.31-7.14(m,3H),7.13-7.03(m,1H),6.95-6.85(m,2H),6.13( d,J=1.0Hz,0.5H),6.01(d,J=0.9Hz,0.5H),3.96(s,1H),3.86(s,1.5H),3.81(t ,J=5.2Hz,4H),3.57(s,1.5H),3.32(s,3H),3.14(q,J=4.8Hz,4H),2.24(s,3H).

[0159] Example 17 Synthesis of 2-(1,5-dimethyl-3-phenyl-1H-pyrrol-2-yl)-N-(3-fluoro-4-(4-(5-fluoropyrimidin-2-yl)piperazin-1-yl)phenyl)-2-(methoxyimino)acetamide (I-9)

[0160]

[0161] Referring to the synthesis operation of Example 16, compound IIc (2.g, 7.74mmol) was condensed with compound IIIb (2.3g, 7.56mmol) to obtain compound I-9 (2.95g, yield 73.1%). LC-MS (ESI, m / z) 546.2 [M+H] + ; 1 H NMR (400 MHz, DMSO-d 6)δ10.32(s,1H),8.48(s,2H),7.50-7.28(m,2H),7.26-7.18(m,2H),7.15(t,J=7.6Hz,1H),7.11-6.90(m,2H),6.14(s,0.5H),5. 99(s,0.5H),3.98(s,1H),3.87(s,1.5H),3.83(t,J=5.0Hz,4H),3.59(s,1.5H),3.31(s,3H),3.00(q,J=5.0Hz,4H),2.25(s,3H).

[0162] Example 18 Synthesis of 2-(1,5-dimethyl-3-phenyl-1H-pyrrol-2-yl)-2-(ethoxyimino)-N-(4-(4-(5-fluoropyrimidin-2-yl)piperazin-1-yl)phenyl)acetamide (I-10)

[0163]

[0164] Referring to the synthesis operation of Example 15, compound I-1 (1.0 g, 2.01 mmol) was reacted with O-ethylhydroxylamine hydrochloride (431 mg, 4.41 mmol) to obtain compound I-10 (695 mg, yield 64.0%), LC-MS (ESI, m / z) 542.3 [M+H] + ; 1 HNMR (400 MHz, DMSO-d 6 )δ10.07(s,0.2H),10.01(s,0.8H),8.48(d,J=0.9Hz,2H),7.49–7.42(m,2H),7.39–7.15(m,4H), 7.09(tt,J=6.3,1.8Hz,1H),6.96–6.86(m,2H),6.13(d,J=1.1Hz,0.8H),6.01(d,J=1.1Hz,0.2H), 4.21(q,J=7.0Hz,1.6H),4.11(q,J=7.0Hz,0.4H),3.81(t,J=5.2Hz,4H),3.55(s,0.6H),3.32(s,2 .4H),3.14(q,J=4.9Hz,4H),2.26–2.22(s,3H),1.15(t,J=7.1Hz,0.6H),1.13(t,J=7.1Hz,2.4H).

[0165] Example 19 Synthesis of 2-(1,5-dimethyl-3-phenyl-1H-pyrrol-2-yl)-2-(allylimino)-N-(4-(4-(5-fluoropyrimidin-2-yl)piperazin-1-yl)phenyl)acetamide (I-11)

[0166]

[0167] Referring to the synthesis operation of Example 15, compound I-1 (1.0 g, 2.01 mmol) was reacted with O-ethylhydroxylamine hydrochloride (431 mg, 4.41 mmol) to obtain compound I-11 (695 mg, yield 64.0%). LC-MS (ESI, m / z) 554.3 [M+H] + ; 1 HNMR (400MHz, CDCl 3 )δ8.37(s,0.5H),8.22(s,2H),7.95(s,0.5H),7.36-7.42(m,2H),7.26-7.30(m,2H),7.2 2-7.24(m,1H),7.11-7.17(m,2H),6.91(s,1H),6.18(s,0.5H),6.06(s,0.5H),5.98-6.06 (m,0.5H),5.74-5.84(m,0.5H),5.17-5.37(m,2H),4.74(d,J=6.0Hz,1H),4.67(s,1H),3. 95(s,4H),3.57-3.59(m,1.5H),3.38-3.40(m,1.5H),3.21(s,4H),2.27(d,J=3.6Hz,3H).

[0168] Example 20 Synthesis of N-(4-(4-(5-fluoropyrimidin-2-yl)piperazin-1-yl)phenyl)-2-(hydroxyimino)-2-(5-methyl-3-phenylthiophen-2-yl)acetamide (I-12)

[0169]

[0170] Referring to the synthesis operation of Example 13, compound I-3 (1.0 g, 2.0 mmol) was reacted with hydroxylamine hydrochloride (401 mg, 4.12 mmol) to obtain compound I-12 (561 mg, yield 53.2%). LC-MS (ESI, m / z) 517.2 [M+H] + ; 1HNMR(400MHz,DMSO-d6)δ12.26(s,0.77H),11.62(s,0.23H),10.11(s,0.20H),9.96(s,0.79H),8.48(s,0.2H),7.35-7.32(m,2H),7.28-7.12(m ,5.3H),6.98(d,J=1.2Hz,0.8H),6.86-6.82(m,2.2H),6.69-6.64(m,0. 2H),3.82-3.79(m,4H),3.13-3.11(m,4H),2.49(s,2.2H),2.45(s,0.8H)

[0171] Example 21 Synthesis of N-(4-(4-(5-fluoropyrimidin-2-yl)piperazin-1-yl)phenyl)-2-(methoxyimino)-2-(5-methyl-3-phenylthiophen-2-yl)acetamide (I-13)

[0172]

[0173] Referring to the synthesis operation of Example 15, compound I-3 (1.0 g, 2.0 mmol) was reacted with O-methylhydroxylamine hydrochloride (401 mg, 4.12 mmol) to obtain compound I-13 (561 mg, yield 53.2%), LC-MS (ESI, m / z) 531.2 [M+H] + ; 1 HNMR (400 MHz, DMSO-d 6 )δ10.16(s,0.33H),10.6(s,0.59H),8.48(s,1.8H),7.33-7.07(m,7.2H),6.98(d,J=0.8Hz,0.6H),6.86-6.8 0(m,2.3H),3.95(s,1.88H),3.82(s,1.14H),3.82-3.79(m4H),3.14-3.12(m,4H),2.49(s,2H),2.46(s,1H).

[0174] Example 22 Synthesis of N-(3-fluoro-4-(4-(5-fluoropyrimidin-2-yl)piperazin-1-yl)phenyl)-2-(hydroxyimino)-2-(5-methyl-3-phenylthiophen-2-yl)acetamide (I-14)

[0175]

[0176] Referring to the synthesis operation of Example 13, compound I-4 was reacted with hydroxylamine hydrochloride to obtain compound I-14. LC-MS (ESI, m / z) 535.2 [M+H] +; 1 HNMR(400MHz,DMSO-d6)δ12.26(s,0.77H),11.62(s,0.23H),10.11(s,0.20H),9.96(s,0.79H),8.48(s,0.2H),7.35-7.32(m,2H),7.28-7.12(m, 5.3H),6.98(d,J=1.2Hz,0.8H),6.86-6.82(m,2.2H),6.69-6.64(m,0.2 H),3.82-3.79(m,4H),3.13-3.11(m,4H),2.49(s,2.2H),2.45(s,0.8H).

[0177] Example 23 Synthesis of N-(3-fluoro-4-(4-(5-fluoropyrimidin-2-yl)piperazin-1-yl)phenyl)-2-(methoxyimino)-2-(5-methyl-3-phenylthiophen-2-yl)acetamide (I-15)

[0178]

[0179] Referring to the synthesis operation of Example 15, compound I-4 (1.0 g, 1.93 mmol) was reacted with O-methylhydroxylamine hydrochloride (401 mg, 4.12 mmol) to obtain compound I-15 (846 mg, yield 80.0%), LC-MS (ESI, m / z) 549.2 [M+H] + ; 1 HNMR(400MHz,DMSO-d6)δ10.16(s,0.33H),10.6(s,0.59H),8.48(s,1.8H),7.33-7.07(m,7.2H),6.98(d,J=0.8Hz,0.6H) ,6.86-6.80(m,2.3H),3.95(s,1.88H),3.82(s,1.14H),3.82-3.79(m,4H),3.14-3.12(m,4H),2.49(s,2H),2.46(s,1H).

[0180] Example 24 Synthesis of N-(3-fluoro-4-(4-(5-fluoropyrimidin-2-yl)piperazin-1-yl)phenyl)-2-(hydroxyimino)-2-(5-methyl-3-phenylfuran-2-yl)acetamide (I-16)

[0181]

[0182] Referring to the synthesis operation of Example 13, compound I-5 (0.97 g, 2.00 mmol) was reacted with hydroxylamine hydrochloride (306 mg, 4.41 mmol) to obtain compound I-16 (782 mg, yield 78.0%). LC-MS (ESI, m / z) 501.2 [M+H] + ; 1 H-NMR (400MHz, CDCl 3 ): δ12.25(s,0.7H),11.57(s,0.3H),10.33(s,0.3H),10.10(s,0.7H),8. 48(s,2H),7.50-7.55(m,2H),7.28-7.43(m,4H),7.21-7.25(m,1H),6.93 -6.95(m,2H),6.55(d,J=0.8Hz,0.7H),6.41(d,J=0.8Hz,0.3H),3.81-3.84(m,4H),3.15-3.17(m,4H),2.07(s,2.1H),1.99(s,0.9H).

[0183] Example 25 Synthesis of N-(3-fluoro-4-(4-(5-fluoropyrimidin-2-yl)piperazin-1-yl)phenyl)-2-(methoxyimino)-2-(5-methyl-3-phenylfuran-2-yl)acetamide (I-17)

[0184]

[0185] Referring to the synthesis operation of Example 15, compound I-5 (1.0 g, 1.93 mmol) was reacted with O-methylhydroxylamine hydrochloride (401 mg, 4.12 mmol) to obtain compound I-17 (846 mg, yield 80.0%), LC-MS (ESI, m / z) 515.2 [M+H] + ; 1 HNMR (400 MHz, DMSO-d 6 )δ10.35(s,0.6H),10.22(s,0.4H),8.48(s,2H),7.52-7.46(m,2H),7.34-7.22(m,5H),6.96-6.89(m,2H),6.5 9(d,J=0.8Hz,0.4H),6.44(d,J=0.8Hz,0.6H),3.84(s,3H),3.83-3.78(m,4H),3.18-3.13(m,4H),2.33(s,3H).

[0186] Effect Example 1 Evaluation of in vitro antifungal activity of some compounds of the present invention

[0187] 1. Strains. The strains used in this experiment were provided by WuXi AppTec, and their information is shown in Table 1.

[0188] Table 1. Experimental test strains

[0189] Serial number Latin name Chinese name serial number 1 Aspergillus flavus Aspergillus flavus ATCC MYA-1004 2 Aspergillus fumigatus Aspergillus fumigatus ATCC-MYA-4609 3 Aspergillus Niger Aspergillus niger CMCC(B)98003 4 Aspergillus terreus Aspergillus terreus ATCC MYA-3633 5 C.albicans Candida albicans ATCC 2876

[0190] 2. Experimental method (refer to CLSI standard test)

[0191] 2.1 Compound test plate preparation

[0192] Accurately weigh 1 to 5 mg of the compound into a sterile Eppendorf tube. Dissolve the compound in DMSO to prepare a high-concentration stock solution, such as 6.4 mg / mL. Depending on the storage requirements of the compound, it is generally stored at -20°C for a long time.

[0193] On the day of the test, thaw and vortex to ensure homogeneity. Perform a 2-fold gradient dilution in DMSO to prepare a 100X compound gradient dilution solution, with a total of 11 gradients. Take 2ul and add it to a 96-well round-bottom plate to obtain a compound test plate.

[0194] 2.2 Preparation of inoculum

[0195] 2.2.1 Candida albicans: Glycerol culture was plated on SDA or PDA plates and cultured overnight. On the day of the test, a single colony was picked from the plate and the turbidity was adjusted to 0.5 McFarland in sterile saline, and then diluted to 5×10 in RPMI 1640 (0.165 M MOPS, pH 7.0) test medium. 2 ~2.5×10 3 CFU / ml, and obtain the inoculum.

[0196] 2.2.1 Aspergillus: Glycerol culture was placed on SDA or PDA plates and cultured for 2 to 7 days to produce spores. On the day of the test, Aspergillus spores were collected from the plates and counted, suspended in 0.1% Tween 20 sterile saline, and the actual number of spores was manually counted using a cell counter. Then, the spores were diluted to 0.2 to 2.5 x 10 using RPMI 1640 (0.165 M MOPS, pH 7.0) test medium. 4 spores / ml to obtain the inoculum.

[0197] 3.3MIC test

[0198] 198 μL of inoculum was added to each well of the compound testing plate.

[0199] 1) Candida albicans: After incubation for 24 hours at 35°C, growth was assessed by monitoring the optical density at 485 nm of each well. The MIC was defined as the lowest drug concentration that inhibited 100% growth compared to the growth control.

[0200] 2) Aspergillus: After incubation for 48 hours at 35°C, growth was assessed by monitoring the optical density at 485 nm of each well. The MIC was defined as the lowest drug concentration that inhibited 100% growth compared to the growth control.

[0201] The experimental results are shown in Table 2.

[0202] Table 2 Minimum inhibitory concentration (μg / mL) of some compounds of the present invention against Aspergillus fumigatus and Candida albicans

[0203]

[0204] The results showed that under the existing test conditions, by comparing the MIC values, some of the compounds of the present invention and the positive control olorofim had the same or better antibacterial activity.

[0205] Effect Example 2 Human Liver Microsome Stability Experiment

[0206] 1. Preparation of test compound and control working solutions:

[0207] 1.1. Working solution: 5 μL of compound and control standard solutions (10 mM, dissolved in dimethyl sulfoxide (DMSO)) were diluted with 495 μL of acetonitrile (ACN) (intermediate solution concentration: 100 μM, 99% ACN).

[0208] 2. Preparation of NADPH coenzyme:

[0209] 2.1. Materials: NADPH powder: β-nicotinamide adenine dinucleotide phosphate reduced form, tetrasodium salt; NADPH·4Na (supplier: BONTAC, catalog number: BT04)

[0210] 2.2. Preparation steps: Weigh an appropriate amount of NADPH powder and dilute it into 2 mM MgCl2 solution (working solution concentration: 2 mM; final concentration in the reaction system: 1 mM).

[0211] 3. Preparation of liver microsomes:

[0212] 2.3.1. Materials:

[0213]

[0214] 2.3.2. Preparation steps: Prepare the microsome working solution of appropriate concentration in 100 mM potassium phosphate buffer. 4. Preparation of stop solution:

[0215] Cold (4°C) acetonitrile (ACN) containing 250 nM tolbutamide and 250 nM labetalol as internal standard (IS) was used as stop solution.

[0216] 5. Test steps:

[0217] Using the Apricot automated workstation, 2 μL / well of compound working solution was added to all 96-well reaction plates, except blank wells (T0, T60, and NCF60). Using the Apricot automated workstation, 100 μL / well of microsome solution was added to all reaction plates (blank, T0, T60, and NCF60). All reaction plates containing compound and microsome mixtures were preincubated at 37°C for 10 minutes. Using the Apricot automated workstation, 98 μL / well of 100 mM potassium phosphate buffer was added to the reaction plate NCF60. The reaction plate NCF60 was incubated at 37°C and timer 1 was started. After preincubation, 98 μL / well of NADPH was added to each reaction plate (blank, T0, and T60) except NCF60 using the Apricot automated workstation to start the reaction. The reaction plates were incubated at 37°C and timer 2 was started.

[0218] At the appropriate stop time point, 600 μL / well of stop solution was added to each reaction plate using the Apricot automated workstation to stop the reaction. Each reaction plate was sealed and shaken for 10 minutes. After shaking, each reaction plate was centrifuged at 4000 rpm and 4°C for 20 minutes. After centrifugation, 300 μL of supernatant was transferred from each reaction plate to eight new 96-well plates using the Apricot automated workstation for LC-MS / MS analysis.

[0219] 6. Data Analysis

[0220] The half-life (T 1 / 2 ) and intrinsic clearance (CL int(mic) )(μL / min / mg):

[0221] Table 3 Stability data of example compounds in human liver microsomes

[0222]

[0223] The results showed that under the existing test conditions, compared with the positive control olorofim (I-1), the example compound I-2 of the present invention had a prolonged human liver microsome metabolic half-life and a slower clearance rate.

[0224] The above embodiments of the present invention are merely examples for clearly illustrating the present invention, and are not intended to limit the embodiments of the present invention. For those skilled in the art, other different forms of changes or modifications can be made based on the above description. It is not necessary and impossible to list all the embodiments here. Any modifications, equivalent substitutions and improvements made within the spirit and principles of the present invention should be included in the protection scope of the claims of the present invention.

Claims

1. A compound represented by formula (I) or a pharmaceutically acceptable salt thereof: in, Y is oxygen, sulfur or NR 1 ; R 1 is hydrogen, C1-C 10 Alkyl or one or more R 1-1 Substituted C1-C 10 alkyl; Each R 1-1 each independently is deuterium or a halogen; R 2 is hydrogen or halogen; X is oxygen or N-OR 3 ; R 3 is hydrogen, -C(O)C1-C 10 Alkyl, C1-C 10 Alkyl, C 2- C 10 Alkenyl or C1-C 10 alkyl; R 4 is a halogen; When X is oxygen, R 1 is hydrogen or is replaced by one or more R 1-1 Substituted C1-C 10 alkyl.

2. The compound of formula (I) or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that: It meets one or more of the following conditions: (1)R 1 In the above, C1-C 10 Alkyl and said one or more R 1-1 Substituted C1-C 10 C1-C 10 The alkyl groups are each independently C1-C6 alkyl groups; the C1-C6 alkyl groups are preferably methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl or tert-butyl groups, and more preferably methyl groups; (2)R 1-1 wherein the halogen is fluorine, chlorine, bromine or iodine, preferably fluorine; (3)R 2 wherein the halogen is fluorine, chlorine, bromine or iodine, preferably fluorine; (4)R 3 In the above-C(O)C1-C 10 C1-C 10 Alkyl, the C1-C 10 The alkyl group and the C1-C 10 C1-C 10 The alkyl groups are each independently C1-C6 alkyl groups; the C1-C6 alkyl groups are preferably methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl or tert-butyl groups, more preferably methyl, ethyl, isopropyl or tert-butyl groups; (5)R 3 In the C 2- C 10 Alkenyl is C 2- C6 alkenyl, the C 2- The C6 alkenyl group is preferably a vinyl group, More preferably (6)R 3 In the case of C1-C 10 The halogen in the alkyl group is fluorine, chlorine, bromine or iodine, preferably fluorine; (7)R 4 In the above, the halogen is fluorine, chlorine, bromine or iodine, preferably fluorine.

3. The compound of formula (I) or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that: The compound represented by formula (I) is a compound represented by formula (IA) or (IB): Y, R 2 and R 3 The definition as in claim 1.

4. The compound of formula (I) or a pharmaceutically acceptable salt thereof according to claim 3, characterized in that: It satisfies one or both of the following conditions: (1) In formula (IA), Y is oxygen, sulfur or NR 1 ; R 1 is hydrogen, C1-C6 alkyl or is replaced by one or more R 1-1 Substituted C1-C6 alkyl; Each R 1-1 each independently is deuterium or a halogen; R 2 is hydrogen or halogen; R 3 is hydrogen, -C(O)C1-C6 alkyl, C1-C6 alkyl or C 2- C6 alkenyl; (2) In formula (IB), Y is oxygen, sulfur or NR 1 ; R 1 is hydrogen or is replaced by one or more R 1-1 Substituted C1-C6 alkyl; Each R 1-1 Each independently is deuterium; R 2 is hydrogen or halogen.

5. The compound of formula (I) or a pharmaceutically acceptable salt thereof according to claim 4, characterized in that: It satisfies one or both of the following conditions: (1) In formula (IA), Y is oxygen, sulfur or N-CH3; R 2 is hydrogen or -F; R 3 For hydrogen, -CH3, -CH2CH3, (2) In formula (IB), Y is oxygen, sulfur or N-CD3; R 2 It is hydrogen or -F.

6. The compound of formula (I) or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 5, characterized in that: The compound represented by formula (I) is any of the following compounds:

7. A compound as shown in formula (II); in, X and Y are as defined in any one of claims 1-6.

8. The compound as shown in formula (II) according to claim 7, characterized in that The compound represented by formula (II) is any of the following compounds:

9. A pharmaceutical composition, characterized in that It comprises a compound as shown in formula (I) according to any one of claims 1 to 6 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

10. Use of a substance A in the preparation of a drug for preventing and / or treating a disease caused by a fungal infection, wherein the fungus is preferably Aspergillus and / or Candida albicans; the Aspergillus is preferably Aspergillus flavus, Aspergillus fumigatus, Aspergillus niger or Aspergillus terreus; The substance A is a compound as shown in formula (I) or a pharmaceutically acceptable salt thereof as described in any one of claims 1 to 6, or a pharmaceutical composition as described in claim 9.

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