MPC inhibitor, and preparation method therefor and use thereof

Abstract

Provided are an MPC inhibitor, and a preparation method therefor and a use thereof. Specifically disclosed is a compound represented by formula (I) or a pharmaceutically acceptable salt thereof. The compound has good physical and chemical properties, has a good inhibitory effect on MPC, has good in vivo and in vitro treatment effects on hair loss, and has low potential toxic and side effects.

Description

An MPC inhibitor, its preparation method and its application

[0001] This application claims priority to Chinese patent application 2024119925277, filed on December 31, 2024. The entire contents of the aforementioned Chinese patent application are incorporated herein by reference. Technical Field

[0002] This invention relates to an MPC inhibitor, its preparation method, and its application. Background Technology

[0003] Hair loss, also known as alopecia or baldness, is a common health problem. Studies have found that by age 50, 50% of men and 25% of women will experience hair loss. Common types of hair loss include androgenetic alopecia (AGA), female pattern hair loss (FPHL), alopecia areata (AA), and thinning hair known as the telogen phase. While hair loss is not a life-threatening disease, it can cause feelings of inferiority, frustration, and decreased well-being, and may even lead to suicide. Currently, three drugs—minoxidil, finasteride, and baricitinib—are approved by the FDA for hair regrowth. All three drugs have been reported to be effective in only 30%-40% of patients after repeated treatment, and have significant toxic side effects, such as increased heart rate, arrhythmia, and weight gain (J Eur Acad Dermatol Venereol. 2022 Feb; 36(2):286-294; J Am Acad Dermatol. 2021 Mar; 84(3):737-746). Therefore, there is an urgent clinical need for highly effective, low-toxicity, and novel mechanisms of action drugs for treating hair loss.

[0004] Hair follicle growth is cyclical. Each cycle consists of a long growth phase (anagen), a short transition phase (catagen), and a short resting phase (telogen). The ability of a hair follicle to maintain this cycle depends on the presence of hair follicle stem cells, or HFSCs. HFSCs are usually inactive, but they can quickly "wake up" and actively divide when a new hair growth cycle begins. Hair loss occurs when HFSCs fail to activate. Literature reports that HFSC activation can be achieved by stimulating the activity of lactate dehydrogenase (LDH), an enzyme that catalyzes the reduction of pyruvate to lactate. Studies have found that inhibition of mitochondrial pyruvate carriers (MPCs) can lead to increased LDH activity in HFSCs. When MPCs are knocked out or inhibited by drugs, pyruvate in the cytoplasm cannot enter the mitochondria, but is instead converted into other metabolites, such as lactate, by lactate dehydrogenase (LDH), thereby increasing LDH activity and activating HFSCs, which can ultimately promote hair growth (Exp Dermatol. 2021 Apr; 30(4):448-456). Currently, MPCs have been shown to be a very promising therapeutic target for treating hair loss.

[0005] UK-5099 was the first MPC inhibitor to be proven to have hair growth activity (J Med Chem. 2021 Feb 25; 64(4):2046-2063). Its mechanism of action is similar to Mpc1 gene knockout (Mpc1 is a transport protein required for pyruvate to enter mitochondria, and its loss of function increases the conversion of pyruvate to lactate). It can block pyruvate from entering mitochondria, thereby promoting lactate production. Increasing lactate production during the metabolism of hair follicle stem cells can activate hair follicle stem cells in the resting phase, thereby promoting hair follicles to enter a new growth cycle.

[0006] In 2024, Pelage Pharmaceuticals (https: / / pelagepharma.com / ) announced that its Phase 2a clinical trial of the MPC inhibitor small molecule PP405 had completed dosing of the first patients, aiming to evaluate the safety and efficacy of PP405 in treating androgenetic alopecia. Early results from the Phase 1 clinical trial showed that PP405 was well-tolerated, and the activation of hair follicle stem cells was statistically significant. As of now, the specific structure of PP405 has not been disclosed.

[0007] Given the limited number of structural types of MPC inhibitors with therapeutic effects on hair loss, with only PP405 currently in clinical trials, there is an urgent need to discover novel inhibitor molecules that possess novel structures, favorable physicochemical properties, good in vitro and in vivo therapeutic effects on hair loss, and low potential toxicity. Summary of the Invention

[0008] The technical problem this invention aims to solve is to overcome the limited variety of MPC inhibitors in the existing technology, and to provide an MPC inhibitor, its preparation method, and its application. The MPC inhibitor provided by this invention has one or more of the following advantages: novel structure, simple preparation, good physicochemical properties (e.g., good compound solubility), good inhibitory effect on MPC, good in vitro and in vivo hair loss treatment effect, and low potential toxicity.

[0009] This invention provides a compound of formula (I) or a pharmaceutically acceptable salt thereof:

[0010] in,

[0011] X is -C(=O)- or -S(=O)2-;

[0012] Y is either N or CH;

[0013] R is hydrogen, halogen, cyano, C1-C6 alkyl, C1-C6 alkoxy, or is surrounded by one or more R. 0-1 Substituted C1-C6 alkyl groups, or those with one or more R 0-2 Substituted C1-C6 alkoxy groups; when there are multiple substituents, they may be the same or different;

[0014] R 0-1 and R 0-2 It is independently C1-C6 alkyl, halogen, or amino;

[0015] R 1 It is a 5-10 membered heterocyclic alkyl, 6-10 membered aryl, 5-10 membered heteroaryl, and is bound by one or more R 1-1 The 6-10 aryl group is replaced, or is replaced by one or more R groups. 1-2 Substituted 5-10 heteroaryl groups; the 5-10 heteroaryl group, substituted with one or more R groups 1-2 In the substituted 5-10 membered heteroaryl and 5-10 membered heterocyclic alkyl groups, the heteroatoms are independently selected from one, two, or three of N, O, and S, and the number of heteroatoms is one, two, or three; when there are multiple substituents, they may be the same or different;

[0016] R 1-1 and R 1-2 It is independently a halogen, amino, C1-C6 alkyl, C1-C6 alkoxy, or a C1-C6 alkyl substituted with one or more halogens; when there are multiple substituents, they may be the same or different;

[0017] R 2 -COOR 2-1 ;

[0018] R 2-1It is hydrogen, C1-C6 alkyl, or composed of one or more R 2-1-1 Substituted C1-C6 alkyl groups;

[0019] R 2-1-1 It is independently C1-C6 alkyl, hydroxyl, halogen, or amino;

[0020] R 3 It is a cyano group.

[0021] In some embodiments, certain groups in the compound of formula (I) or its pharmaceutically acceptable salt are defined as follows, and groups not mentioned are as described in any embodiment of the invention (hereinafter referred to as "in some embodiments"), R, R 0-1 R 1-1 and R 1-2 In this context, the halogen is independently fluorine, chlorine, bromine, or iodine, preferably fluorine, chlorine, or bromine.

[0022] In some implementations, R 1-1 and R 1-2 In this context, the C1-C6 alkoxy groups are independently methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, or tert-butoxy, preferably methoxy, ethoxy, or n-butoxy.

[0023] In some implementations, R, R 1-1 R 1-2 and R 2-1 In this context, the C1-C6 alkyl group, being formed by one or more R... 0-1 The C1-C6 alkyl group substituted with substituted C1-C6 alkyl group and the C1-C6 alkyl group substituted with one or more halogens are independently methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl or tert-butyl, preferably methyl or ethyl.

[0024] In some implementations, R 1 In the context, the 6-10 aryl group and one or more R 1-1 The 6-10 aryl group in the substituted aryl group is independently phenyl or naphthyl, preferably phenyl.

[0025] In some implementations, R 1 In the context, the 5-10 membered heteroaryl group and one or more R groups 1-2 The 5-10-membered heteroaryl group in the substituted 5-10-membered heteroaryl group is independently a 5-6-membered heteroaryl group, wherein the heteroatom in the 5-6-membered heteroaryl group is N, and the number of heteroatoms is 1 or 2; preferably.

[0026] In some embodiments, R is independently a halogen, a C1-C6 alkyl group, or is affected by one or more R groups. 0-1Substituted C1-C6 alkyl groups; R 0-1 Halogens are independent of each other.

[0027] In some implementations, R 1 It is a 6-10 aryl, 5-10 heteroaryl, or surrounded by one or more R 1-1 The 6-10 aryl group is replaced, or is replaced by one or more R groups. 1-2 Substituted 5-10 heteroaryl groups.

[0028] In some implementations, R 1-1 and R 1-2 It is independently a halogen, a C1-C6 alkyl, a C1-C6 alkoxy, or a C1-C6 alkyl substituted with one or more halogens.

[0029] In some implementations, R 2 -COOR 2-1 R 2-1 It is hydrogen or C1-C6 alkyl.

[0030] In some implementations, X is -C (=O)- or -S (=O)2-;

[0031] Y is either N or CH;

[0032] R is a halogen, a C1-C6 alkyl group, or is formed by one or more R. 0-1 Substituted C1-C6 alkyl groups; when there are multiple substituents, they may be the same or different;

[0033] R 0-1 Halogens are independent of each other;

[0034] R 1 It is a 6-10 aryl, 5-10 heteroaryl, or surrounded by one or more R 1-1 The 6-10 aryl group is replaced, or is replaced by one or more R groups. 1-2 Substituted 5-10 heteroaryl groups; the 5-10 heteroaryl group is reacted with one or more R groups. 1-2 In the substituted 5-10 membered heteroaryl groups, the heteroatoms are independently selected from one, two, or three of N, O, and S, and the number of heteroatoms is one, two, or three; when there are multiple substituents, they may be the same or different;

[0035] R 1-1 and R 1-2 It is independently a halogen, a C1-C6 alkyl, a C1-C6 alkoxy, or a C1-C6 alkyl substituted with one or more halogens; when there are multiple substituents, they may be the same or different;

[0036] R 2 -COOR 2-1 ;

[0037] R 2-1 It is hydrogen or C1-C6 alkyl;

[0038] R 3 It is a cyano group.

[0039] In some implementations, X is -C (=O)- or -S (=O)2-;

[0040] Y is either N or CH;

[0041] R is a halogen, a C1-C6 alkyl group, or is formed by one or more R. 0-1 Substituted C1-C6 alkyl groups; when there are multiple substituents, they may be the same or different;

[0042] R 0-1 Halogens are independent of each other;

[0043] R 1 It is pyridyl or is composed of one or more R groups. 1-2 Substituted pyridinyl group;

[0044] R 1-2 It is a halogen on its own; when there are multiple substituents, they may be the same or different;

[0045] R 2 -COOR 2-1 ;

[0046] R 2-1 It is hydrogen or C1-C6 alkyl;

[0047] R 3 It is a cyano group.

[0048] In some implementations, R is fluorine or -CF3.

[0049] In some implementations, R 1 for

[0050] In some implementations... Independently

[0051] In some embodiments, the compound represented by formula (I) is a compound represented by formula (IA) or a compound represented by formula (IB):

[0052] Among them, R, R 1 R 2 The definitions of Y are as described above.

[0053] In some embodiments, the compound represented by formula (I) is any of the following compounds:

[0054] Table 1

[0055] The present invention provides a method for preparing a compound as shown in formula (I) as described above, wherein the preparation method is either Scheme 1 or Scheme 2 as follows:

[0056] Option 1: In an organic solvent and in the presence of an organic base, the compound shown in formula (II) is subjected to a substitution reaction with the compound shown in formula (Int-I) to obtain the compound shown in formula (I):

[0057] Among them, R 2 It is a -COOC1-C6 alkyl group, M is a halogen, and X, Y, R, R 1 and R 3 The definition is as stated above;

[0058] Option 2: In a solvent, in the presence of an inorganic base, the compound shown in formula (III) is hydrolyzed to obtain the compound shown in formula (I):

[0059] Among them, R 2 For -COOH, X, Y, R, R 1 and R 3 The definition is as described above.

[0060] The present invention provides a pharmaceutical composition comprising a compound as shown in formula (I) as described above, or a pharmaceutically acceptable salt thereof, and at least one pharmaceutical excipient.

[0061] The present invention provides the use of a compound of formula (I) as described above, or a pharmaceutically acceptable salt thereof, in the preparation of an MPC inhibitor.

[0062] In some embodiments, the MPC inhibitor can be used in mammalian organisms or in vitro (e.g., for experimental purposes, as a standard or control sample for comparison, or to prepare a kit according to conventional methods in the art to provide rapid detection of the effect of MPC inhibition).

[0063] The present invention provides the use of a compound of formula (I) as described above or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition as described above, in the preparation of a medicament for treating or preventing MPC-mediated diseases.

[0064] In some implementations, the MPC-mediated disease is hair loss, diabetes, metabolic disease, or inflammation.

[0065] The present invention provides the use of the compound shown in formula (I) above or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition thereof, in the preparation of a medicament for promoting lactate release.

[0066] The present invention provides the use of the compound shown in formula (I) above or a pharmaceutically acceptable salt thereof, or the use of the above pharmaceutical composition in the preparation of a medicament for treating hair loss.

[0067] Terminology Explanation

[0068] In this invention, the term "pharmaceutically acceptable salt" refers to a salt obtained by reacting a compound with a pharmaceutically acceptable acid or base. When a 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 a 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. See Handbook of Pharmaceutical Salts: Properties, Selection, and Use (P. Heinrich Stahl, Camille G. Wermuth, 2011, 2nd Revised Edition) for details.

[0069] The term "ester" includes physiologically hydrolyzable esters (compounds of the present invention that can be hydrolyzed under physiological conditions to release free acids or alcohols). Alternatively, the compounds of the present invention may themselves be esters.

[0070] In this invention, the structural segments This refers to the structural segment being connected to the rest of the molecule via this bond. For example, This refers to the pyridinyl group through It is connected to the rest of the molecule.

[0071] In this invention, the term "one or more" refers to 1, 2, 3, 4, 5 or 6, for example, 1, 2 or 3.

[0072] In this invention, the term "halogen" refers to fluorine, chlorine, bromine, or iodine.

[0073] In this invention, the term "alkyl" refers to a saturated monovalent hydrocarbon group that has a specified number of carbon atoms (e.g., C1-C6), is straight-chain or branched. Alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, n-hexyl, etc.

[0074] In this invention, the term "alkylene" refers to a saturated divalent hydrocarbon group obtained by removing two hydrogen atoms from a saturated straight-chain or branched hydrocarbon group. Alkylenes include, but are not limited to: methylene (-CH2-), ethylene {including -CH2CH2- or -CH(CH3)-}, isopropylene {including -CH(CH3)CH2- or -C(CH3)2-}, etc.

[0075] In this invention, the term "alkoxy" refers to the group R. Y -O-,R Y The definition is the same as the term "alkyl". Alkoxy groups include, but are not limited to: methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, sec-butoxy, tert-butoxy, n-pentoxy, n-hexoxy, etc.

[0076] In this invention, the term "aryl" refers to an aryl group having a specified number of carbon atoms (e.g., C6-C). 10 Aromatic groups are cyclic, unsaturated monovalent hydrocarbon groups, which can be monocyclic or polycyclic (e.g., two). In the case of polycyclic groups, the monocyclic rings share two atoms and one bond, and each ring is aromatic. The aryl group is connected to the rest of the molecule through an aromatic ring. Aromatic groups include, but are not limited to, phenyl and naphthyl groups.

[0077] In this invention, the term "heteroaryl" refers to a cyclic, unsaturated group having a specified number of ring atoms (e.g., 5-6), a specified number of heteroatoms (e.g., 0, 1, 2, or 3), and a specified type of heteroatom (one or more of N, O, and S), and possessing aromaticity; the heteroaryl group is attached to the rest of the molecule via a carbon atom or a heteroatom. Heteroaryl groups include, but are not limited to, those with aromaticity. wait.

[0078] In this invention, the term "heterocyclic alkyl" refers to a cyclic, saturated monovalent group having a specified number of ring atoms (e.g., 3-12, 4-8, or 5-6), a specified number of heteroatoms (e.g., 1, 2, or 3), and a specified type of heteroatom (one or more of N, O, and S), wherein the heterocyclic alkyl group is attached to the rest of the molecule by a carbon atom or a heteroatom.

[0079] Without violating common sense in the field, the above-mentioned preferred conditions can be combined arbitrarily to obtain various preferred embodiments of the present invention.

[0080] The reagents and raw materials used in this invention are all commercially available.

[0081] The positive and progressive effects of this invention are as follows: Compared with the prior art, the compounds disclosed in this invention have one or more of the following advantages:

[0082] (1) Novel structure;

[0083] (2) Good physicochemical properties (e.g., good solubility of the compound);

[0084] (3) It has a good inhibitory effect on MPC;

[0085] (4) Most of them showed good lactate release-promoting effects and had good in vivo and in vitro hair loss treatment effects;

[0086] (5) Low potential toxic side effects.

[0087] Furthermore, the MPC inhibitor preparation method provided by this invention has simple conditions, readily available raw materials, low preparation cost, and high product yield. Detailed Implementation

[0088] The present invention is further illustrated below by way of embodiments, but the invention is not limited to the scope of the embodiments described herein. Experimental methods in the following embodiments that do not specify specific conditions were performed according to conventional methods and conditions, or as selected according to the product instructions.

[0089] Example 1: (E)-3-(1-(6-chloronicotinyl)-4-fluoro-1H-indol-3-yl)-2-cyanoacrylate (Compound I-1)

[0090] Add the starting material Int.I-1-a (0.2 g, 0.774 mmol) to a 50 mL round-bottom flask and dissolve it in 8 mL of dichloromethane. Add DMAP (9.5 mg, 0.077 mmol) and triethylamine (0.118 g, 1.16 mmol), and add a 5 mL dichloromethane solution of the starting material Int.I-1-b (0.163 g, 0.929 mmol) at 0 °C. After 30 min, allow the mixture to return to room temperature and react for 2 h at room temperature. Quench the reaction with 1 N hydrochloric acid, extract with dichloromethane (3 x 20 mL), collect the organic phase and evaporate to dryness to obtain the crude product. Recrystallize the crude product to give compound I-1.

[0091] ESI-MS (m / z): 398.06 [M+H] +

[0092] 1H NMR (400MHz, DMSO-d6) δ9.02(d,J=2.0Hz,1H),8.45(s,1H),8.03(dd,J=7.8,2.0Hz,1H),7.89(s,1H),7.71(dd,J=6.4,1.2Hz, 1H),7.49(ddd,J=7.3,6.4,5.0Hz,1H),7.40(d,J=7.8Hz,1H),7.23–7.14(m,1H),4.30(q,J=7.1Hz,2H),1.31(t,J=7.1Hz,3H).

[0093] Example 2: (E)-2-cyano-3-(4-fluoro-1-(2-methoxynicotinyl)-1H-indol-3-yl)ethyl acrylate (compound I-2)

[0094] Compound I-2 was prepared using the same synthetic method as compound I-1.

[0095] ESI-MS (m / z): 394.11 [M+H] +

[0096] 1 H NMR (400MHz, DMSO-d6) δ8.47(s,1H),8.28(dd,J=4.5,1.8Hz,1H),8.07(dd,J=7.1,1.9Hz,1H),7.81(s,1H),7.73(dd,J=6.5, 1.1Hz, 1H), 7.50 (ddd, J=7.5, 6.4, 5.1Hz, 1H), 7.22–7.16 (m, 2H), 4.30 (q, J=7.1Hz, 2H), 3.99 (s, 3H), 1.31 (t, J=7.1Hz, 3H).

[0097] Example 3: (E)-3-(1-(5-bromonicotinyl)-4-fluoro-1H-indol-3-yl)-2-cyanoacrylate (Compound I-3)

[0098] Compound I-3 was prepared using the same synthetic method as compound I-1.

[0099] ESI-MS (m / z): 442.01 [M+H] +

[0100] 1H NMR (400MHz, DMSO-d6) δ9.07(t,J=1.7Hz,1H),8.66(t,J=1.7Hz,1H),8.47(s,1H),8.40(t,J=1.7Hz,1H),7.82(s,1H),7.71(dd,J= 6.4, 1.3Hz, 1H), 7.49 (ddd, J=7.3, 6.4, 5.0Hz, 1H), 7.19 (ddd, J=8.3, 7.4, 1.1Hz, 1H), 4.30 (q, J=7.1Hz, 2H), 1.31 (t, J=7.1Hz, 3H).

[0101] Example 4: (E)-3-(1-(2-chloronicotinyl)-4-fluoro-1H-indol-3-yl)-2-cyanoacrylate (Compound I-4)

[0102] Compound I-4 was prepared using the same synthetic method as compound I-1.

[0103] ESI-MS (m / z): 398.06 [M+H] +

[0104] 1 H NMR (400MHz, DMSO-d6) δ8.47(dd,J=4.4,1.9Hz,1H),8.41(s,1H),8.05(dd,J=6.8,1.9Hz,1H),7.88(s,1H),7.72(dd,J=6.3,1.2Hz,1H),7.5 7(dd,J=6.9,4.3Hz,1H), 7.50(ddd,J=7.5,6.3,5.0Hz,1H), 7.19(ddd,J=8.3,7.4,1.2Hz,1H), 4.30(q,J=7.1Hz,2H), 1.31(t,J=7.1Hz,3H).

[0105] Example 5: (E)-3-(1-benzoyl-4-fluoro-1H-indol-3-yl)-2-cyanoacrylate (Compound I-5)

[0106] Compound I-5 was prepared using the same synthetic method as compound I-1.

[0107] ESI-MS (m / z): 363.11 [M+H] +

[0108] 1H NMR (400MHz, DMSO-d6) δ8.45(s,1H),8.14(s,1H),7.95(dd,J=6.3,2.2Hz,2H),7.71(dd,J=6.4, 1.3Hz,1H),7.53–7.46(m,4H),7.22–7.15(m,1H),4.30(q,J=7.1Hz,2H),1.31(t,J=7.1Hz,3H).

[0109] Example 6: (E)-2-cyano-3-(1-(4-ethoxybenzoyl)-4-fluoro-1H-indol-3-yl)ethyl acrylate (compound I-6)

[0110] Compound I-6 was prepared using the same synthetic method as compound I-1.

[0111] ESI-MS (m / z): 407.13 [M+H] +

[0112] 1 H NMR (400MHz, DMSO-d6) δ8.46(s,1H),8.11(s,1H),7.94–7.89(m,2H),7.71(dd,J=6.4,1.3Hz,1H),7.49(ddd,J=7.3,6.4,5.0Hz,1H),7.19( ddd,J=8.3,7.4,1.1Hz,1H),7.03–6.99(m,2H),4.30(q,J=7.1Hz,2H),4.03(q,J=6.6Hz,2H),1.39(t,J=6.7Hz,3H),1.31(t,J=7.1Hz,3H).

[0113] Example 7: (E)-3-(1-(4-butoxybenzoyl)-4-fluoro-1H-indol-3-yl)-2-cyanoacrylate (Compound I-7)

[0114] Compound I-7 was prepared using the same synthetic method as compound I-1.

[0115] ESI-MS (m / z): 435.16 [M+H] +

[0116] 1H NMR(400MHz,DMSO-d6)δ8.49(s,1H),8.11(s,1H),7.93–7.89(m,2H),7.71(dd,J =6.4,1.3Hz,1H),7.49(ddd,J=7.3,6.4,5.0Hz,1H),7.19(ddd,J=8.3,7.4,1.1Hz ,1H),7.02–6.96(m,2H),4.30(q,J=7.1Hz,2H),3.97(t,J=6.3Hz,2H),1.77(p,J =6.6Hz, 2H), 1.50 (h, J = 6.9Hz, 2H), 1.31 (t, J = 7.1Hz, 3H), 0.97 (t, J = 6.9Hz, 3H).

[0117] Example 8: (E)-2-cyano-3-(1-(4-ethylbenzoyl)-4-fluoro-1H-indole-3-yl)ethyl acrylate (Compound I-8)

[0118] Compound I-8 was prepared using the same synthetic method as compound I-1.

[0119] ESI-MS (m / z): 391.14 [M+H] +

[0120] 1 H NMR (400MHz, DMSO-d6) δ8.50(s,1H),8.11(s,1H),7.88–7.81(m,2H),7.71(dd,J=6.4,1.3Hz,1H),7.49(ddd,J=7.3,6.4,5.0Hz,1H),7.32(dt,J=7 .9,1.1Hz,2H),7.19(ddd,J=8.3,7.4,1.1Hz,1H),4.30(q,J=7.1Hz,2H), 2.69(qt,J=7.3,1.0Hz,2H), 1.31(t,J=7.1Hz,3H), 1.23(t,J=7.3Hz,3H).

[0121] Example 9: (E)-2-cyano-3-(4-fluoro-1-pyridyl-1H-indol-3-yl)ethyl acrylate (compound I-9)

[0122] Compound I-9 was prepared using the same synthetic method as compound I-1.

[0123] ESI-MS (m / z): 364.10 [M+H] +

[0124] 1 H NMR(400MHz, DMSO-d6)δ8.74(dd,J=4.3,1.7Hz,1H),8.41(s,1H),8.02(dd,J= 6.7,1.5Hz,1H),7.94(td,J=7.1,1.7Hz,1H),7.74(dd,J=6.4,1.3Hz,1H),7.62 (ddd,J=7.3,4.3,1.6Hz,1H),7.51(ddd,J=7.5,6.3,5.0Hz,1H),7.47(s,1H),7 .19(ddd,J=8.3,7.4,1.2Hz,1H),4.30(q,J=7.1Hz,2H),1.31(t,J=7.1Hz,3H).

[0125] Example 10: (E)-2-cyano-3-(1-pyridyl-4-(trifluoromethyl)-1H-pyrrolo[2,3-b]pyridin-3-yl)ethyl acrylate (Compound I-10)

[0126] Compound I-10 was prepared using the same synthetic method as compound I-1.

[0127] ESI-MS (m / z): 415.09 [M+H] +

[0128] 1 H NMR (400MHz, DMSO-d6) δ8.75(dd,J=4.3,1.7Hz,1H),8.61(d,J=2.7Hz,1H),8.46(s,1H),8.39(s,1H),8.04(dd,J=6.7,1.6Hz,1H), 7.97(td,J=7.0,1.7Hz,1H), 7.77(d,J=2.9Hz,1H), 7.62(ddd,J=7.3,4.2,1.6Hz,1H), 4.30(q,J=7.1Hz,2H), 1.31(t,J=7.1Hz,3H).

[0129] Example 11: (E)-3-(1-(3,5-bis(trifluoromethyl)benzoyl)-4-fluoro-1H-pyrrolo[2,3-b]pyridin-3-yl)-2-cyanoacrylate (Compound I-11)

[0130] Compound I-11 was prepared using the same synthetic method as compound I-1.

[0131] ESI-MS (m / z): 500.08 [M+H] +

[0132] 1 H NMR(400MHz, DMSO-d6)δ8.52(t,J=4.8Hz,1H),8.47(s,1H),8.43(s,1H),8.35(d,J=2.1Hz,2H), 8.20(t,J=2.2Hz,1H),7.25(dd,J=8.0,4.5Hz,1H),4.30(q,J=7.1Hz,2H),1.31(t,J=7.1Hz,3H).

[0133] Example 12: (E)-2-cyano-3-(4-fluoro-1-pyridyl-1H-indole-3-yl)acrylic acid (compound I-12)

[0134] I-9 (1.2 mmol) was added to a 50 mL round-bottom flask, dissolved in 8 mL of tetrahydrofuran, and then excess lithium hydroxide monohydrate (0.50 g, 12.0 mmol) was added. The mixture was hydrolyzed at room temperature for 2 h. After the reaction was completed by TLC monitoring, tetrahydrofuran was removed by vacuum distillation, and about 20 mL of ethyl acetate was added. The mixture was stirred for 30 min, separated, and the pH of the aqueous phase was adjusted to <6. The precipitated solid was filtered and dried to obtain the target compound I-12.

[0135] ESI-MS (m / z): 336.07 [M+H] +

[0136] 1 H NMR (400MHz, DMSO-d6) δ12.59(s,1H),8.74(dd,J=4.3,1.6Hz,1H),8.43(s,1H),8.02(dd,J=6.7,1.5Hz,1H),7.94(td,J=7.1,1.7Hz,1H),7.7 4(dd,J=6.4,1.3Hz,1H),7.62(ddd,J=7.3,4.2,1.6Hz,1H),7.51(ddd,J=7.5,6.3,5.0Hz,1H),7.47(s,1H),7.19(ddd,J=8.3,7.4,1.2Hz,1H).

[0137] Example 13: (E)-3-(1-(6-chloronicotinyl)-4-fluoro-1H-indol-3-yl)-2-cyanoacrylic acid (compound I-13)

[0138] Compound I-13 was prepared using the same synthetic method as compound I-12.

[0139] ESI-MS (m / z): 370.03 [M+H] +

[0140] 1 H NMR (400MHz, DMSO-d6) δ12.53(s,1H),9.02(d,J=2.0Hz,1H),8.43(s,1H),8.03(dd,J=7.8,2.0Hz,1H),7.90(s,1H),7. 71(dd,J=6.4,1.2Hz,1H),7.49(ddd,J=7.3,6.4,5.0Hz,1H),7.40(d,J=7.8Hz,1H),7.19(ddd,J=8.3,7.4,1.1Hz,1H).

[0141] Example 14: (E)-3-(1-(2-chloronicotinyl)-4-fluoro-1H-indol-3-yl)-2-cyanoacrylic acid (compound I-14)

[0142] Compound I-14 was prepared using the same method as compound I-12.

[0143] ESI-MS (m / z): 370.03 [M+H] +

[0144] 1 H NMR (400MHz, DMSO-d6) δ12.58(s,1H),8.47(dd,J=4.4,1.9Hz,1H),8.43(s,1H),8.05(dd,J=6.9,1.9Hz,1H),7.86(s,1H),7. 72(dd,J=6.3,1.2Hz,1H), 7.57(dd,J=7.0,4.3Hz,1H), 7.50(ddd,J=7.5,6.3,5.0Hz,1H), 7.19(ddd,J=8.3,7.4,1.2Hz,1H).

[0145] Example 15: (E)-2-cyano-3-(4-fluoro-1-(2-methoxynicotinyl)-1H-indol-3-yl)acrylic acid (compound I-15)

[0146] Compound I-15 was prepared using the same synthetic method as compound I-12.

[0147] ESI-MS (m / z): 366.08 [M+H] +

[0148] 1H NMR (400MHz, DMSO-d6) δ12.57(s,1H),8.43(s,1H),8.28(dd,J=4.5,1.8Hz,1H),8.07(dd,J=7.1,1.9Hz,1H), 7.83(s,1H),7.73(dd,J=6.5,1.1Hz,1H),7.50(ddd,J=7.5,6.4,5.1Hz,1H),7.22–7.16(m,2H),3.99(s,3H).

[0149] Example 16: (E)-3-(1-(5-bromonicotinyl)-4-fluoro-1H-indol-3-yl)-2-cyanoacrylic acid (compound I-16)

[0150] Compound I-16 was prepared using the same synthetic method as compound I-12.

[0151] ESI-MS (m / z): 413.98 [M+H] +

[0152] 1 H NMR (400MHz, DMSO-d6) δ12.57(s,1H),9.07(t,J=1.7Hz,1H),8.66(t,J=1.7Hz,1H),8.43(s,1H),8.40(t,J=1.7Hz, 1H), 7.89 (s, 1H), 7.71 (dd, J=6.4, 1.2Hz, 1H), 7.49 (ddd, J=7.3, 6.4, 5.0Hz, 1H), 7.19 (ddd, J=8.3, 7.4, 1.1Hz, 1H).

[0153] Example 17: (E)-2-cyano-3-(1-(4-ethoxybenzoyl)-4-fluoro-1H-indol-3-yl)acrylic acid (compound I-17)

[0154] Compound I-17 was prepared using the same synthetic method as compound I-12.

[0155] ESI-MS (m / z): 413.98 [M+H] +

[0156] 1H NMR (400MHz, DMSO-d6) δ12.57(s,1H),8.43(s,1H),8.15(s,1H),7.93–7.89(m,2H),7.71(dd,J=6.4,1.3Hz,1H),7.49(ddd, J=7.3,6.4,5.0Hz,1H),7.19(ddd,J=8.3,7.4,1.1Hz,1H),7.04–6.97(m,2H),4.03(q,J=6.6Hz,2H),1.39(t,J=6.7Hz,3H).

[0157] Example 18: (E)-3-(1-(4-butoxybenzoyl)-4-fluoro-1H-indol-3-yl)-2-cyanoacrylic acid (compound I-18)

[0158] Compound I-18 was prepared using the same method as compound I-12.

[0159] ESI-MS (m / z): 407.13 [M+H] +

[0160] 1 H NMR (400MHz, DMSO-d6) δ12.57(s,1H),8.43(s,1H),8.11(s,1H),7.94–7.89(m,2H),7.71(dd,J=6.4,1.3Hz,1H),7.49(ddd,J=7.3,6.4,5.0Hz,1H) ,7.19(ddd,J=8.3,7.4,1.1Hz,1H),7.02–6.96(m,2H),3.97(t,J=6.3Hz, 2H), 1.77 (p, J = 6.6Hz, 2H), 1.50 (h, J = 6.9Hz, 2H), 0.97 (t, J = 6.9Hz, 3H).

[0161] Example 19: (E)-2-cyano-3-(1-pyridyl-4-(trifluoromethyl)-1H-pyrrolo[2,3-b]pyridin-3-yl)acrylic acid (compound I-19)

[0162] Compound I-19 was prepared using the same synthetic method as compound I-12.

[0163] ESI-MS (m / z): 387.06 [M+H] +

[0164] 1H NMR (400MHz, DMSO-d6) δ12.60(s,1H),8.75(dd,J=4.3,1.7Hz,1H),8.61(d,J=2.7Hz,1H),8.43(s,1H),8.38(s,1H) ,8.04(dd,J=6.7,1.6Hz,1H),7.97(td,J=7.0,1.7Hz,1H),7.77(d,J=2.9Hz,1H),7.62(ddd,J=7.3,4.3,1.6Hz,1H).

[0165] Example 20: (E)-3-(1-(3,5-bis(trifluoromethyl)benzoyl)-4-fluoro-1H-pyrrolo[2,3-b]pyridin-3-yl)-2-cyanoacrylic acid (compound I-20)

[0166] Compound I-20 was prepared using the same method as compound I-12.

[0167] ESI-MS (m / z): 472.05 [M+H] +

[0168] 1 H NMR (400MHz, DMSO-d6) δ12.59(s,1H),8.52(t,J=4.8Hz,1H),8.44(s,1H),8.43( s,1H),8.35(d,J=2.1Hz,2H),8.20(t,J=2.2Hz,1H),7.25(dd,J=8.0,4.5Hz,1H).

[0169] Example 21: (E)-3-(1-((6-chloropyridin-3-yl)sulfonyl)-4-fluoro-1H-indol-3-yl)-2-cyanoacrylate (Compound I-21)

[0170] Add 50 mg (0.194 mmol) of raw material Int.I-1-a to a round-bottom flask, dissolve in 1 mL of acetonitrile, add DMAP (1.18 mg, 0.0097 mmol) and DIPEA (25 mg, 0.194 mmol), and after 10 min, add 0.25 mL of an acetonitrile solution of raw material Int.I-1-c (0.290 mmol). Heat in an oil bath at 40 °C for 4 h. Reduce the acetonitrile concentration under reduced pressure, extract with saturated brine and dichloromethane, collect the organic phase and evaporate to dryness to obtain the crude product. Dissolve the crude product in dichloromethane, remove water with anhydrous sodium sulfate, and recrystallize to give 40 mg of a yellow-green solid.

[0171] ESI-MS (m / z): 434.03 [M+H]+

[0172] 1 H NMR (400MHz, DMSO-d6) δ8.88(d,J=1.9Hz,1H),8.47(d,J=0.6Hz,2H),8.09(dd,J=8.6,1.9Hz,1H),7.92(dd,J=6.3,1.1Hz,1H) ,7.50(dd,J=7.5,6.3Hz,1H),7.41(d,J=8.6Hz,1H),7.22(dd,J=7.5,1.1Hz,1H),4.28(q,J=7.1Hz,2H),1.31(t,J=7.1Hz,3H).

[0173] Example 22: (E)-2-cyano-3-(4-fluoro-1-((2-methoxypyridin-3-yl)sulfonyl)-1H-indol-3-yl)ethyl acrylate (compound I-22)

[0174] Compound I-22 was prepared using the same method as compound I-21.

[0175] ESI-MS (m / z): 430.08 [M+H] +

[0176] 1 H NMR (400MHz, DMSO-d6) δ8.52(d,J=0.6Hz,1H),8.47(s,1H),8.25–8.15(m,2H),7.92(dd,J=6.3,1.1Hz,1H),7.50(dd,J= 7.5, 6.3Hz, 1H), 7.41 (dd, J=7.6, 4.1Hz, 1H), 7.22 (dd, J=7.5, 1.2Hz, 1H), 4.28 (q, J=7.1Hz, 2H), 1.31 (t, J=7.1Hz, 3H).

[0177] Example 23: (E)-3-(1-((5-bromopyridin-3-yl)sulfonyl)-4-fluoro-1H-indol-3-yl)-2-cyanoacrylate (Compound I-23)

[0178] Compound I-23 was prepared using the same synthetic method as compound I-21.

[0179] ESI-MS (m / z): 477.98 [M+H] +

[0180] 1H NMR (400MHz, DMSO-d6) δ8.89(t,J=1.7Hz,1H),8.56(t,J=1.7Hz,1H),8.50–8.45(m,2H),8.11(t,J=1.7Hz,1H),7.92(dd ,J=6.3,1.1Hz,1H),7.50(dd,J=7.5,6.3Hz,1H),7.22(dd,J=7.5,1.1Hz,1H),4.33–4.25(m,2H),1.31(t,J=7.1Hz,3H).

[0181] Example 24: (E)-3-(1-((2-chloropyridin-3-yl)sulfonyl)-4-fluoro-1H-indol-3-yl)-2-cyanoacrylate (Compound I-24)

[0182] Compound I-24 was prepared using the same synthetic method as compound I-21.

[0183] ESI-MS (m / z): 434.03 [M+H] +

[0184] 1 H NMR (400MHz, DMSO-d6) δ8.52(d,J=0.6Hz,1H),8.47(s,1H),8.38(dd,J=4.0,1.8Hz,1H),8.14(dd,J=7.5,1.9Hz,1H), 7.92(dd,J=6.3,1.2Hz,1H),7.54–7.42(m,2H),7.22(dd,J=7.5,1.2Hz,1H),4.33–4.25(m,2H),1.31(t,J=7.1Hz,3H).

[0185] Example 25: (E)-2-cyano-3-(4-fluoro-1-(benzenesulfonyl)-1H-indol-3-yl)ethyl acrylate (Compound I-25)

[0186] Compound I-25 was prepared using the same synthetic method as compound I-21.

[0187] ESI-MS (m / z): 399.07 [M+H] +

[0188] 1H NMR (400MHz, DMSO-d6) δ8.51–8.45(m,2H),7.94(dd,J=6.3,1.1Hz,1H),7.84–7.76(m,2H),7.69–7. 60(m,1H),7.54–7.44(m,3H),7.22(dd,J=7.5,1.1Hz,1H),4.33–4.25(m,2H),1.31(t,J=7.1Hz,3H).

[0189] Example 26: (E)-2-cyano-3-(1-((4-ethoxyphenyl)sulfonyl)-4-fluoro-1H-indol-3-yl)ethyl acrylate (Compound I-26)

[0190] Compound I-26 was prepared using the same synthetic method as compound I-21.

[0191] ESI-MS (m / z): 443.10 [M+H] +

[0192] 1 H NMR (400MHz, DMSO-d6) δ8.47(d,J=3.2Hz,2H),7.94(dd,J=6.3,1.2Hz,1H),7.80–7.72(m,2H),7.50(dd,J=7.5,6.3Hz,1H),7.22(d d,J=7.5,1.2Hz,1H),7.04–6.96(m,2H),4.28(q,J=7.1Hz,2H),4.03(t,J=6.7Hz,2H),1.40(t,J=6.7Hz,3H),1.31(t,J=7.1Hz,3H).

[0193] Example 27: (E)-3-(1-((4-butoxyphenyl)sulfonyl)-4-fluoro-1H-indol-3-yl)-2-cyanoacrylate (Compound I-27)

[0194] Compound I-27 was prepared using the same synthetic method as compound I-21.

[0195] ESI-MS (m / z): 471.13 [M+H] +

[0196] 1H NMR(400MHz,DMSO-d6)δ8.47(d,J=3.2Hz,2H),7.94(dd,J=6.3,1.2Hz,1H),7 .80–7.72(m,2H),7.50(dd,J=7.5,6.3Hz,1H),7.22(dd,J=7.5,1.2Hz,1H),7. 04–6.96(m,2H),4.28(q,J=7.1Hz,2H),3.97(d,J=12.7Hz,1H),1.77(p,J=6. 7Hz, 2H), 1.49 (h, J = 6.9Hz, 2H), 1.31 (t, J = 7.1Hz, 3H), 0.97 (t, J = 7.0Hz, 3H).

[0197] Example 28: (E)-2-cyano-3-(1-((4-ethylphenyl)sulfonyl)-4-fluoro-1H-indol-3-yl)ethyl acrylate (Compound I-28)

[0198] Compound I-28 was prepared using the same synthetic method as compound I-21.

[0199] ESI-MS (m / z): 427.10 [M+H] +

[0200] 1 H NMR (400MHz, DMSO-d6) δ8.51–8.45(m,2H),7.94(dd,J=6.3,1.1Hz,1H),7.78–7.71(m,2H),7.50(dd,J=7.5,6.3Hz,1H),7.35–7 .27(m,2H),7.22(dd,J=7.5,1.2Hz,1H),4.28(q,J=7.1Hz,2H),2.73–2.63(m,2H),1.31(t,J=7.1Hz,3H),1.23(t,J=7.2Hz,3H).

[0201] Example 29: (E)-2-cyano-3-(4-fluoro-1-(pyridin-2-ylsulfonyl)-1H-indol-3-yl)ethyl acrylate (compound I-29)

[0202] Compound I-29 was prepared using the same synthetic method as compound I-21.

[0203] ESI-MS (m / z): 400.07 [M+H] +

[0204] 1H NMR (400MHz, DMSO-d6) δ8.72(s,1H),8.49–8.41(m,2H),7.95(dd,J=6.3,1.2Hz,1H),7.93–7.85(m,2H),7.50(dd,J=7. 5,6.3Hz,1H),7.33(ddd,J=5.4,4.2,3.3Hz,1H),7.22(dd,J=7.5,1.1Hz,1H),4.33–4.25(m,2H),1.31(t,J=7.1Hz,3H).

[0205] Example 30: (E)-2-cyano-3-(1-(pyridin-2-ylsulfonyl)-4-(trifluoromethyl)-1H-indol-3-yl)ethyl acrylate (Compound I-30)

[0206] Compound I-30 was prepared using the same synthetic method as compound I-21.

[0207] ESI-MS (m / z): 450.07 [M+H] +

[0208] 1 H NMR (400MHz, DMSO-d6) δ8.77(d,J=0.6Hz,1H),8.50(d,J=0.6Hz,1H),8.47–8.41(m,1H),8.02(dd,J=6.1,1.3Hz,1H),7.94–7.85(m,2H), 7.78(dd,J=10.1,6.1Hz,1H),7.66(dd,J=10.1,1.2Hz,1H),7.33(ddd,J=5.4,4.2,3.3Hz,1H),4.33–4.25(m,2H),1.31(t,J=7.1Hz,3H).

[0209] Example 31: (E)-3-(1-((3,5-bis(trifluoromethyl)phenyl)sulfonyl)-4-fluoro-1H-indol-3-yl)-2-cyanoacrylate (Compound I-31)

[0210] Compound I-31 was prepared using the same synthetic method as compound I-21.

[0211] ESI-MS (m / z): 535.05 [M+H] +

[0212] 1H NMR (400MHz, DMSO-d6) δ8.47(d,J=3.2Hz,2H),8.16(d,J=2.2Hz,2H),7.94(dd,J=6.3,1.2Hz,1H),7.83(t,J=2 .2Hz,1H),7.50(dd,J=7.5,6.3Hz,1H),7.22(dd,J=7.5,1.2Hz,1H),4.33–4.25(m,2H),1.31(t,J=7.1Hz,3H).

[0213] Example 32: (E)-2-cyano-3-(4-fluoro-1-(pyridin-2-ylsulfonyl)-1H-indol-3-yl)ethyl acrylate (compound I-32)

[0214] Compound I-32 was prepared using the same method as compound I-12.

[0215] ESI-MS (m / z): 372.04 [M+H] +

[0216] 1 H NMR(400MHz,DMSO-d6)δ8.72(d,J=0.6Hz,1H),8.54(s,1H),8.47–8.41(m,1H),7.95(dd,J=6.3,1.2Hz,1H),7 .93–7.85(m,2H),7.50(dd,J=7.5,6.3Hz,1H),7.33(ddd,J=5.4,4.2,3.3Hz,1H),7.22(dd,J=7.5,1.1Hz,1H).

[0217] Example 33: (E)-3-(1-((6-chloropyridin-3-yl)sulfonyl)-4-fluoro-1H-indol-3-yl)-2-cyanoacrylic acid (Compound I-33)

[0218] Compound I-33 was prepared using the same synthetic method as compound I-12.

[0219] ESI-MS (m / z): 406.00 [M+H] +

[0220] 1H NMR (400MHz, DMSO-d6) δ8.88(d,J=1.9Hz,2H),8.54(s,2H),8.47(d,J=0.6Hz,2H),8.09(dd,J=8.6,1.9Hz,2H) ,7.92(dd,J=6.3,1.2Hz,2H),7.50(dd,J=7.5,6.3Hz,2H),7.41(d,J=8.6Hz,2H),7.22(dd,J=7.5,1.1Hz,2H).

[0221] Example 34: (E)-3-(1-((2-chloropyridin-3-yl)sulfonyl)-4-fluoro-1H-indol-3-yl)-2-cyanoacrylic acid (compound I-34)

[0222] Compound I-34 was prepared using the same synthetic method as compound I-12.

[0223] ESI-MS (m / z): 400.07 [M+H] +

[0224] 1 H NMR(400MHz, DMSO-d6)δ8.56–8.50(m,2H),8.38(dd,J=4.0,1.8Hz,1H),8.14(dd,J=7.5, 1.9Hz,1H),7.92(dd,J=6.3,1.1Hz,1H),7.54–7.42(m,2H),7.22(dd,J=7.5,1.1Hz,1H).

[0225] Example 35: (E)-2-cyano-3-(4-fluoro-1-((2-methoxypyridin-3-yl)sulfonyl)-1H-indol-3-yl)acrylic acid (compound I-35)

[0226] Compound I-35 was prepared using the same method as compound I-12.

[0227] ESI-MS (m / z): 400.07 [M+H] +

[0228] 1 H NMR(400MHz,DMSO-d6)δ8.53(d,J=8.4Hz,2H),8.25–8.15(m,2H),7.92(dd,J=6.3,1.1Hz, 1H), 7.50 (dd, J=7.5, 6.3Hz, 1H), 7.41 (dd, J=7.6, 4.1Hz, 1H), 7.22 (dd, J=7.5, 1.2Hz, 1H).

[0229] Example 36: (E)-3-(1-((5-bromopyridin-3-yl)sulfonyl)-4-fluoro-1H-indol-3-yl)-2-cyanoacrylic acid (Compound I-36)

[0230] Compound I-36 was prepared using the same synthetic method as compound I-12.

[0231] ESI-MS (m / z): 449.95 [M+H] +

[0232] 1 H NMR(400MHz, DMSO-d6)δ8.89(t,J=1.7Hz,1H),8.59–8.52(m,2H),8.48(s,1H),8.11(t,J=1.7 Hz,1H),7.92(dd,J=6.3,1.2Hz,1H),7.50(dd,J=7.5,6.3Hz,1H),7.22(dd,J=7.5,1.1Hz,1H).

[0233] Example 37: (E)-2-cyano-3-(1-((4-ethoxyphenyl)sulfonyl)-4-fluoro-1H-indole-3-yl)acrylic acid (compound I-37)

[0234] Compound I-37 was prepared using the same synthetic method as compound I-12.

[0235] ESI-MS (m / z): 415.07 [M+H] +

[0236] 1 H NMR (400MHz, DMSO-d6) δ8.54(s,1H),8.49(d,J=0.6Hz,1H),7.94(dd,J=6.3,1.1Hz,1H),7.80–7.72(m,2H),7.50(d d,J=7.5,6.3Hz,1H),7.22(dd,J=7.5,1.2Hz,1H),7.04–6.96(m,2H),4.03(t,J=6.7Hz,2H),1.40(t,J=6.7Hz,3H).

[0237] Example 38: (E)-3-(1-((4-butoxyphenyl)sulfonyl)-4-fluoro-1H-indol-3-yl)-2-cyanoacrylic acid (compound I-38)

[0238] Compound I-38 was prepared using the same synthetic method as compound I-12.

[0239] ESI-MS (m / z): 443.10 [M+H] +

[0240] 1 H NMR(400MHz, DMSO-d6)δ8.54(s,1H),8.49(d,J=0.6Hz,1H),7.94(dd,J=6.3,1.1Hz,1H),7.80–7.72(m,2H),7.50(dd,J=7.5,6.3Hz,1H),7 .22(dd,J=7.5,1.1Hz,1H),7.04–6.96(m,2H),3.97(d,J=12.7Hz,1H),1.77(p,J=6.7Hz,2H),1.49(h,J=6.9Hz,2H),0.97(t,J=7.0Hz,3H).

[0241] Example 39: (E)-2-cyano-3-(1-(pyridin-2-ylsulfonyl)-4-(trifluoromethyl)-1H-indol-3-yl)acrylic acid (compound I-39)

[0242] Compound I-39 was prepared using the same synthetic method as compound I-12.

[0243] ESI-MS (m / z): 422.03 [M+H] +

[0244] 1 H NMR(400MHz, DMSO-d6)δ8.77(s,1H),8.48–8.41(m,2H),8.02(dd,J=6.1,1.3Hz,1H),7.94–7.85(m, 2H), 7.78 (dd, J=10.1, 6.1Hz, 1H), 7.66 (dd, J=10.1, 1.1Hz, 1H), 7.33 (ddd, J=5.4, 4.2, 3.3Hz, 1H).

[0245] Example 40: (E)-3-(1-((3,5-bis(trifluoromethyl)phenyl)sulfonyl)-4-fluoro-1H-indol-3-yl)-2-cyanoacrylic acid (Compound I-40)

[0246] Compound I-40 was prepared using the same method as compound I-12.

[0247] ESI-MS (m / z): 507.02 [M+H] +

[0248] 1 H NMR (400MHz, DMSO-d6) δ8.54(s,1H),8.48(d,J=0.6Hz,1H),8.16(d,J=2.2Hz,2H),7.94(dd,J=6 .3,1.1Hz,1H),7.83(t,J=2.2Hz,1H),7.50(dd,J=7.5,6.3Hz,1H),7.22(dd,J=7.5,1.1Hz,1H).

[0249] Example 1: In vitro lactate release experiment of the compound

[0250] 1) Instruments and reagents

[0251] The experimental equipment and main reagents for the in vitro lactic acid release experiment are as follows (see Table 2):

[0252] Table 2

[0253] The main experimental reagents for the in vitro lactic acid release experiment are shown in Table 3:

[0254] Table 3

[0255] 2) Experimental Operation

[0256] Step 1: MCF-10A cell passage

[0257] (1) Remove the original culture medium;

[0258] (2) Add about 2 mL of PBS, gently shake the culture flask to rinse the cells, and then aspirate and discard the PBS.

[0259] (3) Add about 1 mL of trypsin and gently shake the culture flask to infiltrate all cells;

[0260] Note: MCF-10A cells are relatively difficult to digest. Please extend the digestion time until the cells shrink and become round. Gently tap the side of the culture flask and the cells can slide off. This digestion can be stopped at this time. It usually takes 10 to 15 minutes.

[0261] (4) Place the culture flask in an incubator for digestion. When you see the cells in the middle of the cell block become obviously rounded and have gaps under a microscope, you can stop the process. Do not tap the culture flask throughout the process.

[0262] (5) Add 3 mL of serum-containing culture medium to stop digestion, pipette the cells to detach them from the cell wall, and repeatedly pipette the cells in the liquid to make them as much as possible a single-cell suspension.

[0263] (6) Collect the cell suspension and centrifuge at 1200 rpm / min for 3 minutes. After centrifugation, aspirate and discard the supernatant.

[0264] (7) Add fresh culture medium, blow a few times to mix the cells, inoculate into new culture flasks according to the ratio, replenish the culture medium, loosen the bottle cap or use a breathable bottle cap for culture.

[0265] Step 2: Determining the maximum lactate release rate concentration of UK-5099

[0266] (1) Cells were treated with the positive control drug UK-5099, and the amount of LA released was detected. Different concentrations were set: 0, 2.5, 5, 10, 20, 50, and 100 μM.

[0267] (2) MCF-10A cells were cultured and, after reaching confluence, were digested and collected. The cells were then cultured at a concentration of 1×10⁻⁶ cells / cells. 6 Cells were seeded at a rate of 100 cells / dish in 6cm culture dishes and cultured overnight for 12 hours. Then, different concentrations of UK-5099 were added, with each concentration repeated twice, and the mixture was cultured for 36 hours.

[0268] (3) Take out the cells to be tested, add 0.8 mL of extraction solution one, and lyse at room temperature for 30 min. Centrifuge at 12000 g for 10 min at 4℃, take 0.8 mL of supernatant, add 0.15 mL of extraction solution two, centrifuge at 12000 g for 10 min at 4℃, and take the supernatant for testing.

[0269] (4) Sample addition table (see Table 4):

[0270] Table 4

[0271] (5) Calculated based on standard products

[0272] LA content (mM / 10) 6 cell) = (A 测定 -A 空白 ) / (A 标准 -A 空白 )×C 标准

[0273] C 标准 =Concentration of the standard, 3mM

[0274] The obtained data were processed, and the concentration at which the positive control drug UK-5099 achieved the maximum lactic acid release rate was calculated to be 10 μM.

[0275] Step 3: Lactic acid (LA) content detection

[0276] (1) The concentration of 10 μM at which the maximum lactic acid release rate of the positive drug UK-5099 is achieved.

[0277] (2) MCF-10A cells were cultured until they reached confluence. The cells were then digested and collected at a concentration of 1×10⁻⁶.6 Cells were seeded at a rate of 100 cells / dish in 6cm culture dishes and cultured overnight for 12 hours. Then, 10μM of different compounds were added and the cells were cultured for 36 hours.

[0278] (3) Take out the cells to be tested, add 0.8 mL of extraction solution one, and lyse at room temperature for 30 min. Centrifuge at 12000 g for 10 min at 4℃, take 0.8 mL of supernatant, add 0.15 mL of extraction solution two, centrifuge at 12000 g for 10 min at 4℃, and take the supernatant for testing.

[0279] (4) Sample addition table (see Table 5):

[0280] Table 5

[0281] (5) Calculate the relative expression level of LA in cells after the addition of the compound: take the absorbance of the DMSO group as a unit and calculate the ratio of each group to the DMSO group.

[0282] 3) Results of the compound's lactate release experiment (see Table 6)

[0283] Table 6

[0284] The experimental results shown in Table 6 above indicate that the compound of the present invention has a better or comparable lactate release-promoting effect compared with the positive control (UK-5099).

[0285] Example 2: Compound Solubility Test

[0286] DMSO solutions of some compounds of this invention (I-1, I-4, I-9, I-10, I-16, I-19, I-21, I-24, I-29), compound I-49 from patent CN118459439A, and UK-5099 were successively diluted with methanol to 128, 64, 32, 16, 8, 4, 2, 1.0, 0.5, and 0.25 μg / mL, respectively. Standard curves were plotted using the peak areas of the corresponding concentrations in HPLC. Excess compound was added to an EP tube containing PBS buffer (pH 7.4), sonicated at room temperature for 30 min, allowed to stand for 10 min, and the supernatant was filtered through a filter membrane. The peak area was measured by HPLC and substituted into the standard curve to obtain the solubility values. The solubility test results of the compounds are shown in Table 7.

[0287] Table 7

[0288] * indicates compound I-49 in patent CN118459439A.

[0289] The experimental results shown in Table 7 above indicate that the compound of the present invention has better solubility compared with the positive control (UK-5099) and the compound of patent CN118459439A.

[0290] Example 3: In vitro inhibitory activity test of the compound on normal human cells

[0291] The antiproliferative activity of some compounds of this invention at a single concentration (10 μM) against MRC-5 human normal embryonic lung fibroblasts (cell source: ATCC) was determined, with UK-5099 as a control. Specific results are shown in the table (unit: in h%in 10 μM):

[0292] Test method: Take cells in good logarithmic growth phase and test them at a density of 4 × 10⁻⁶ cells per well. 3 Cells were seeded into 96-well plates, with 8 replicates per group, and cultured in a cell culture incubator. After cell adhesion, the cells were transfected with the corresponding plasmid. After 48 hours of culture, the drug-containing medium was discarded, and 10 μL of freshly prepared CCK8 detection solution (brand: Engreen) was added to each well. The plates were then incubated for another 4 hours, and the OD value was measured at 450 nm using a microplate reader. The experiment was repeated three times, and the average value was taken as the final result. The inhibition rate (%) was calculated. The results are shown in Table 8.

[0293] Table 8

[0294] As can be seen from Table 8, the compounds of the present invention have weaker inhibitory activity against MRC-5 normal human embryonic lung fibroblasts compared to UK-5099, and have lower toxic side effects, revealing that the compounds of the present invention may have lower toxic side effects when used as anti-tumor drugs.

[0295] Example 4: Hair growth experiment of C57BL / 6J mice

[0296] 1) Experimental conditions

[0297] (1) Laboratory animals

[0298] Species: C57BL / 6J mouse, grade: SPF, weight: male mouse 18g (purchased from Shanghai Slack Laboratory Animal Co., Ltd.);

[0299] (2) Environmental adaptation

[0300] Environmental acclimatization for 7 days prior to the experiment;

[0301] (3) Feeding conditions

[0302] Stocking density: 10 individuals / cage (1 compound), cage space displacement frequency: 1 time / week;

[0303] (4) Rearing environment conditions

[0304] Standards for breeding environment conditions: National Standard of the People's Republic of China GB14925-2010;

[0305] (5) Feed

[0306] Feeding method: Free access;

[0307] Standard nutritional components: crude protein, crude fat, crude fiber, crude ash, moisture, calcium, and phosphorus.

[0308] (6) Drinking water

[0309] Type: Drinking water for laboratory animals (autoclaved tap water);

[0310] Water supply method: Water is provided in bottles and can be consumed freely;

[0311] 2) Experimental methods

[0312] (1) Route of administration: Topical application; Volume of administration: 20 μM 300 μL; Dosage: Distilled water, anhydrous ethanol and polyethylene 400 (5:3:2).

[0313] (2) Trial period: The drug was administered once a day and observed for 16 days after administration.

[0314] (3) Observation indicators:

[0315] General symptoms:

[0316] Observe the growth of the mouse's skin and fur daily, record the time it takes for the skin of each mouse's shaved area to change from pink to gray, and from gray to full fur growth, take photos, record the data, and score the data.

[0317] The scoring criteria are as follows (see Table 9):

[0318] Table 9

[0319] System autopsy:

[0320] After 16 days of drug administration, the animals were euthanized by anesthesia and bloodletting with 3% sodium pentobarbital, and their hair growth was examined.

[0321] 3) The experimental results are as follows (see Table 10):

[0322] Table 10

[0323] The experimental results shown in Table 10 above indicate that, compared with the positive control (UK-5099), the compounds of the present invention all exhibit superior hair growth-promoting effects and are all superior to the marketed drug minoxidil.

[0324] Example 5: Experiment on the effect of compounds on hERG potassium channels

[0325] The potential cardiotoxic side effects of some compounds of this invention were preliminarily investigated in vitro using the hERG potassium channel inhibition assay. The experimental procedure is as follows:

[0326] 1) Cell preparation

[0327] CHO-hERG cells (Millipore PrecisION™ hERG-CHO Recombinant Cell Line, Cat#CYL3038) were cultured at 175 cm⁻¹. 2 In the culture flask, when the cell density grows to 60-80%, remove the culture medium, wash once with 7 mL PBS, and then add 3 mL Detachin (Yaji Biotechnology, A1110501) for digestion.

[0328] After complete digestion, add 7 mL of culture medium to neutralize, then centrifuge, aspirate the supernatant, and resuspend in 5 mL of culture medium to ensure a cell density of 2–5 × 10⁻⁶ cells / mL. 6 / mL.

[0329] 2) Electrophysiological recording process

[0330] The single-cell high-impedance sealing and whole-cell pattern formation processes were all automated by the Qpatch instrument. After obtaining the whole-cell recording pattern, the cells were clamped at -80 mV. Before applying a 5-second +20 mV depolarization stimulus, a 50-millisecond -50 mV pre-voltage was applied, followed by repolarization to -50 mV for 5 seconds, and then back to -80 mV. This voltage stimulus was applied every 15 seconds. After recording for 2 minutes, extracellular fluid was applied for another 2 minutes of recording, and then the drug administration process began. The compound concentration started from the lowest test concentration of 0.1 μM, and each test concentration was administered for 2 minutes. After all concentrations were administered, a positive control compound of 10 μM UK-5099 was administered. At least 3 cells (n≥3) were tested for each concentration.

[0331] 3) Compound preparation

[0332] The stock solution of the compound was diluted with extracellular fluid. 2 μL of the stock solution was added to 998 μL of extracellular fluid, and then the solution was serially diluted 5-fold in extracellular fluid containing 0.2% DMSO to obtain the final concentration to be tested. Experimental data were analyzed using XLFit software.

[0333] The experimental results are shown in Table 11:

[0334] Table 11

[0335] The experimental results shown in Table 11 above demonstrate that the compound hERG IC of this invention... 50 The values ​​were all greater than 10 μM, indicating relatively low potential cardiotoxicity.

Claims

1. A compound of formula (I) or a pharmaceutically acceptable salt thereof: in, X is -C(=O)- or -S(=O)2-; Y is either N or CH; R is hydrogen, halogen, cyano, C1-C6 alkyl, C1-C6 alkoxy, or is surrounded by one or more R. 0-1 Substituted C1-C6 alkyl groups, or those with one or more R 0-2 Substituted C1-C6 alkoxy groups; when there are multiple substituents, they may be the same or different; R 0-1 and R 0-2 It is independently C1-C6 alkyl, halogen, or amino; R 1 It is a 5-10 membered heterocyclic alkyl, 6-10 membered aryl, 5-10 membered heteroaryl, and is bound by one or more R 1-1 The 6-10 aryl group is replaced, or is replaced by one or more R groups. 1-2 Substituted 5-10 heteroaryl groups; the 5-10 heteroaryl group, substituted with one or more R groups 1-2 In the substituted 5-10 membered heteroaryl and 5-10 membered heterocyclic alkyl groups, the heteroatoms are independently selected from one, two, or three of N, O, and S, and the number of heteroatoms is one, two, or three; when there are multiple substituents, they may be the same or different; R 1-1 and R 1-2 It is independently a halogen, amino, C1-C6 alkyl, C1-C6 alkoxy, or a C1-C6 alkyl substituted with one or more halogens; when there are multiple substituents, they may be the same or different; R 2 -COOR 2-1 ; R 2-1 It is hydrogen, C1-C6 alkyl, or composed of one or more R 2-1-1 Substituted C1-C6 alkyl groups; R 2-1-1 It is independently C1-C6 alkyl, hydroxyl, halogen, or amino; R 3 It is a cyano group.

2. The compound of formula (I) as claimed in claim 1, characterized in that, The compound of formula (I) or a pharmaceutically acceptable salt thereof satisfies one or more of the following conditions: (1) R, R 0-1 R 1-1 and R 1-2 In this context, the halogen is independently fluorine, chlorine, or bromine; (2)R 1-1 and R 1-2 In this context, the C1-C6 alkoxy groups are independently methoxy, ethoxy, or n-butoxy; (3) R, R 1-1 R 1-2 and R 2-1 In this context, the C1-C6 alkyl group, being formed by one or more R... 0-1 The C1-C6 alkyl group substituted with substituted C1-C6 alkyl group and the C1-C6 alkyl group substituted with one or more halogens are independently methyl or ethyl; (4)R 1 In the context, the 6-10 aryl group and one or more R 1-1 In the substituted 6-10 aryl group, the 6-10 aryl group is independently a phenyl group; (5)R 1 In the context, the 5-10 membered heteroaryl group and one or more R groups 1-2 The 5-10 heteroaryl group in the substituted 5-10 heteroaryl group is independently a 5-6 heteroaryl group, wherein the heteroatom in the 5-6 heteroaryl group is N, and the number of heteroatoms is 1 or 2.

3. The compound of formula (I) as claimed in claim 1, characterized in that, The compound of formula (I) or a pharmaceutically acceptable salt thereof satisfies one or more of the following conditions: (1) R is independently a halogen, a C1-C6 alkyl group, or is contained in one or more R groups. 0-1 Substituted C1-C6 alkyl groups; R 0-1 Halogens are independent of each other; (2)R 1 It is a 6-10 aryl, 5-10 heteroaryl, or surrounded by one or more R 1-1 The 6-10 aryl group is replaced, or is replaced by one or more R groups. 1-2 Substituted 5-10 heteroaryl groups; (3)R 1-1 and R 1-2 It is independently a halogen, a C1-C6 alkyl, a C1-C6 alkoxy, or a C1-C6 alkyl substituted with one or more halogens; (4)R 2 -COOR 2-1 R 2-1 It is hydrogen or C1-C6 alkyl.

4. The compound of formula (I) as claimed in claim 1, characterized in that, X is -C(=O)- or -S(=O)2-; Y is either N or CH; R is a halogen, a C1-C6 alkyl group, or is formed by one or more R. 0-1 Substituted C1-C6 alkyl groups; when there are multiple substituents, they may be the same or different; R 0-1 Halogens are independent of each other; R 1 It is a 6-10 aryl, 5-10 heteroaryl, or surrounded by one or more R 1-1 The 6-10 aryl group is replaced, or is replaced by one or more R groups. 1-2 Substituted 5-10 heteroaryl groups; the 5-10 heteroaryl group is reacted with one or more R groups. 1-2 In the substituted 5-10 membered heteroaryl groups, the heteroatoms are independently selected from one, two, or three of N, O, and S, and the number of heteroatoms is one, two, or three; when there are multiple substituents, they may be the same or different; R 1-1 and R 1-2 It is independently a halogen, a C1-C6 alkyl, a C1-C6 alkoxy, or a C1-C6 alkyl substituted with one or more halogens; when there are multiple substituents, they may be the same or different; R 2 -COOR 2-1 ; R 2-1 It is hydrogen or C1-C6 alkyl; R 3 It is a cyano group.

5. The compound of formula (I) as claimed in claim 1, characterized in that, The compound of formula (I) or a pharmaceutically acceptable salt thereof satisfies one or more of the following conditions: (1) R is fluorine or -CF3; (2)R 1 for (3) Independently 6. The compound of formula (I) as claimed in any one of claims 1-5, or a pharmaceutically acceptable salt thereof, characterized in that, The compound represented by formula (I) is either the compound represented by formula (IA) or the compound represented by formula (IB): Among them, R, R 1 R 2 The definition of Y is as described in any one of claims 1-5.

7. The compound of formula (I) as claimed in claim 1, characterized in that, The compound represented by formula (I) is any of the following compounds:

8. A method for preparing a compound of formula (I) as described in any one of claims 1-7, characterized in that, The preparation method is either Scheme 1 or Scheme 2 as follows: Option 1: In an organic solvent and in the presence of an organic base, the compound shown in formula (II) is subjected to a substitution reaction with the compound shown in formula (Int-I) to obtain the compound shown in formula (I): Among them, R 2 It is a -COOC1-C6 alkyl group, M is a halogen, and X, Y, R, R 1 and R 3 The definition is as described in any one of claims 1-7; Option 2: In a solvent, in the presence of an inorganic base, the compound shown in formula (III) is hydrolyzed to obtain the compound shown in formula (I): Among them, R 2 For -COOH, X, Y, R, R 1 and R 3 The definition is as described in any one of claims 1-7.

9. A pharmaceutical composition comprising a compound of formula (I) as described in any one of claims 1-7 or a pharmaceutically acceptable salt thereof, and at least one pharmaceutical excipient.

10. The use of a compound of formula (I) as described in any one of claims 1-7, or a pharmaceutically acceptable salt thereof, or the use of a pharmaceutical composition as described in claim 9 in the preparation of a medicament for treating or preventing MPC-mediated diseases; said diseases preferably alopecia, diabetes, metabolic diseases, or inflammation.

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