Substituted 3-thiazolidine-2,4-dione Mcl-1 protein inhibitors and preparation methods and applications thereof

Abstract

The present invention discloses a substituted 3-thiazolidine-2,4-dione Mcl-1 protein inhibitor, its preparation method and application. The compound has a structure represented by general formula (I): The compound of the present invention has strong inhibitory activity against Mcl-1 protein and can prevent or treat related mammalian diseases caused by abnormal expression of Mcl-1 protein. The present invention also relates to the pharmaceutical use of a composition containing a compound of general formula I.

Description

Technical Field

[0001] The present invention relates to a substituted 3-thiazolidine-2,4-dione Mcl-1 protein inhibitor, its preparation, pharmaceutical composition and medical use, belonging to the field of pharmaceutical technology. Background Art

[0002] Apoptosis, also known as programmed cell death (PCD), is a conserved and highly regulated process of cell death and an important protective mechanism in multicellular organisms. It can remove non-functional, harmful, mutated and damaged cells in animals, thus protecting them from the uncontrolled cell growth and proliferation caused by DNA mutations, and plays an important role in maintaining the body's homeostasis and embryonic growth and development. Evasion of apoptosis is a hallmark of malignant tumors and one of the reasons for tumorigenesis, development and resistance to various conventional anti-tumor drug treatments. It has been found that there are mainly two apoptotic pathways in apoptosis, namely the death receptor-mediated extrinsic apoptotic pathway and the mitochondria-mediated intrinsic apoptotic pathway. So far, restoring normal apoptosis of tumor cells by acting on key regulators in the apoptotic pathway has become a very promising anti-tumor strategy.

[0003] Studies have shown that the B-cell leukemia / lymphoma-2 (Bcl-2) protein family is a key regulator of apoptosis and plays an important role in the mitochondria-mediated intrinsic pathway. According to their different functions and structures, the Bcl-2 protein family can be divided into three categories: (1) anti-apoptotic proteins, such as Bcl-2, Bcl-xL, Mcl-1, etc.; (2) multi-domain pro-apoptotic proteins, such as Bax, Bak; (3) pro-apoptotic BH3-only proteins, such as Bad, Bim, Noxa, etc. When endogenous apoptosis occurs in cells, BH3-only proteins activate Bax and Bak, causing them to bind to the outer mitochondrial membrane and release pro-apoptotic factors such as cytochrome c and Smac protein into the cell by changing the permeability of the outer membrane, thus activating caspase and causing apoptosis. Anti-apoptotic proteins Bcl-2, Bcl-xL and Mcl-1 can form heterooligomers with pro-apoptotic proteins Bad, Bim, Bax and Bak to inhibit their activities, thus achieving the effect of evading apoptosis. It has been found that anti-apoptotic proteins Bcl-2, Bcl-xL and Mcl-1 are highly expressed in a variety of malignant tumor cells (such as breast cancer, prostate cancer, lymphoma and leukemia, etc.), and are closely related to the occurrence, recurrence and drug resistance of related tumors to chemotherapy drugs. Therefore, the development of small molecule inhibitors to antagonize the activity of anti-apoptotic Bcl-2 protein has become a hot spot in the current research and development of anti-tumor drugs.

[0004] In the past few decades, many small-molecule anti-apoptotic Bcl-2 protein inhibitors with novel structures have been reported successively and shown good anti-tumor activities. In particular, the first selective Bcl-2 inhibitor, Venetoclax / ABT-199, was approved by the US Food and Drug Administration (FDA) for the treatment of patients with chronic lymphocytic leukemia with deletion of chromosome 17p, which greatly encouraged drug researchers. However, ABT-199 has poor binding ability to Mcl-1 and is not ideal in inducing apoptosis of certain hematological diseases and solid tumor cells that rely on Mcl-1 for survival. In view of this clinical reality, searching for novel small-molecule inhibitors targeting Mcl-1 has become a research topic with great challenges and huge application value in the field of current tumor (especially malignant tumor) treatment. Summary of the Invention

[0005] Aiming at the deficiencies of the prior art, the present invention provides a substituted 3-thiazolidine-2,4-dione Mcl-1 protein inhibitor, and the present invention also provides a preparation method of this compound.

[0006] The present invention further provides a pharmaceutical composition and a medical use of this compound.

[0007] The technical solution of the present invention is as follows:

[0008] I. Substituted 3-thiazolidine-2,4-dione Mcl-1 protein inhibitor

[0009] A substituted 3-thiazolidine-2,4-dione Mcl-1 protein inhibitor is a compound having the structure of general formula (I) or a pharmaceutically acceptable salt thereof.

[0010]

[0011] In general formula (I), R1 is an alkyl group, an aryl group, a heteroaryl group; R1 is preferably an optionally substituted C1-C10 alkyl group, a C3-C10 cycloalkyl group, a C5-C15 aryl group, and a monocyclic heteroaryl group containing 5 or 6 ring atoms, or a bicyclic heteroaryl group having 8 to 15 ring atoms, and the heteroaryl group contains 1-4 heteroatoms, and the heteroatoms are independently selected from O, S, N or oxidized S or N; a carbon atom or a nitrogen atom is the connection point of the heteroaromatic ring structure to maintain a stable aromatic ring;

[0012] In general formula (I), R2 is an isopropyl group or -A-R4;

[0013] Among them, A is a CH2, NH, O, or S atom, preferably CH2; R4 is an optionally substituted aryl or heteroaryl; R4 is preferably an optionally substituted C5-C15 aryl, a monocyclic heteroaryl having 5 or 6 ring atoms, or a bicyclic heteroaryl having 8 to 15 ring atoms, and the heteroaryl contains 1-4 heteroatoms, and the heteroatoms are independently selected from O, S, N, or oxidized S or N; a carbon atom or a nitrogen atom is a connection point of the heteroaromatic ring structure, maintaining a stable aromatic ring;

[0014] In the general formula (I), R3 is an optionally substituted aryl or heteroaryl; R3 is preferably an optionally substituted C5-C15 aryl, a monocyclic heteroaryl having 5 or 6 ring atoms, or a bicyclic heteroaryl having 8 to 15 ring atoms, and the heteroaryl contains 1-4 heteroatoms, and the heteroatoms are independently selected from O, S, N, or oxidized S or N; a carbon atom or a nitrogen atom is a connection point of the heteroaromatic ring structure, maintaining a stable aromatic ring;

[0015] The group or substituent is selected from hydroxyl, halogen, nitro, cyano, guanidino, carboxyl, halo C1-C6 alkyl, C1-C6 alkoxy, C1-C6 alkyl, C3-C8 cycloalkyl, C6-C10 aryl, aralkyloxy, a heteroaryl having 5-10 ring atoms containing 1-2 heteroatoms, and 1-3 of the above groups or substituents are connected at any accessible position to produce a stable compound.

[0016] Preferably according to the present invention, in the general formula (I),

[0017] R1 is a halo C1-C6 alkyl, C1-C6 alkyl, C3-C8 cycloalkyl, an aromatic group Ar connected to a morpholine group or a piperazine group substituted or unsubstituted by 1-2 hydroxyl, halogen, nitro, or cyano substituents, -NH-R5; Ar is a phenyl, naphthyl, pyridyl, pyridazinyl, pyrazinyl, indolizinyl, quinazolinyl, purinyl, indolyl, quinolinyl, pyrimidinyl, pyrrolyl, pyrazolyl, thiazolyl, benzothiazolyl, thienyl, benzo[b]thienyl, isoxazolyl, oxadiazolyl, isothiazolyl, tetrazolyl, imidazolyl, triazinyl, furyl, benzofuryl, and indolyl with 1 substituent or without a substituent; R5 is a C1-C6 alkyl substituted or unsubstituted by 1-2 hydroxyl, halogen, nitro, or cyano substituents, and the above aromatic group Ar connected by a C1-C3 alkylene; the substituents are hydroxyl, halogen, nitro, cyano, guanidino, carboxyl, halo C1-C6 alkyl, C1-C6 alkoxy, C1-C6 alkyl, C3-C8 cycloalkyl, C5-C10 aryl, and a heteroaryl having 5-10 ring atoms containing 1-2 heteroatoms;

[0018] R2 is isopropyl or -CH2-R4; R4 is 3-indolyl and phenyl, naphthyl, pyridyl, pyridazinyl, pyrazinyl, indolizinyl, quinazolinyl, purinyl, indolyl, quinolinyl, pyrimidinyl, pyrrolyl, pyrazolyl, thiazolyl, benzothiazolyl, thienyl, benzo[b]thienyl, isoxazolyl, oxadiazolyl, isothiazolyl, tetrazolyl, imidazolyl, triazinyl, furyl, benzofuryl and indolyl which have 1-2 substituents or no substituents; the substituent is hydroxyl, halogen, nitro, cyano, guanidyl, carboxyl, halo C1-C6 alkyl, C1-C6 alkoxy, C1-C6 alkyl, C3-C8 cycloalkyl, C5-C10 aryl, substituted aralkyloxy, heteroaryl having 5-10 ring atoms and containing 1-2 heteroatoms;

[0019] R3 is an aromatic group Ar connected by a morpholine group or a piperazine group which is substituted or unsubstituted by 1-2 hydroxyl, halogen, nitro, cyano substituents, -NH-R5; Ar is phenyl, naphthyl, pyridyl, pyridazinyl, pyrazinyl, indolizinyl, quinazolinyl, purinyl, indolyl, quinolinyl, pyrimidinyl, pyrrolyl, pyrazolyl, thiazolyl, benzothiazolyl, thienyl, benzo[b]thienyl, isoxazolyl, oxadiazolyl, isothiazolyl, tetrazolyl, imidazolyl, triazinyl, furyl, benzofuryl and indolyl which have 1 substituent or no substituents; R4 is C1-C6 alkyl which is substituted or unsubstituted by 1-2 hydroxyl, halogen, nitro, cyano substituents, and the above aromatic group Ar connected by C1-C3 alkylene; the substituent is hydroxyl, halogen, nitro, cyano, guanidyl, carboxyl, halo C1-C6 alkyl, C1-C6 alkoxy, C1-C6 alkyl, C3-C8 cycloalkyl, C5-C10 aryl, heteroaryl having 5-10 ring atoms and containing 1-2 heteroatoms;

[0020] According to the present invention, further preferably, the compound of the above general formula (I) is one of the following:

[0021] (S)-N-(3-Nitro-4-chlorobenzenesulfonyl)-2-(2-(5-((4'-chloro-[1,1'-biphenyl]-4-yl)methylene)-thiazolidine-2,4-dione-3-yl)acetamido)-3-phenylpropanamide (A1)

[0022] (S)-N-(4-Methylbenzenesulfonyl)-2-(2-(5-((4'-chloro-[1,1'-biphenyl]-4-yl)methylene)-thiazolidine-2,4-dione-3-yl)acetamido)-3-phenylpropanamide (A2)

[0023] (S)-N-(4-chlorobenzenesulfonyl)-2-(2-(5-((4'-chloro-[1,1'-biphenyl]-4-yl)methylene)-thiazolidine-2,4-dione-3-yl)acetamido)-3-phenylpropanamide (A3)

[0024] (S)-N-(3-nitro-4-chlorobenzenesulfonyl)-2-(2-(5-((4'-methyl-[1,1'-biphenyl]-4-yl)methylene)-thiazolidine-2,4-dione-3-yl)acetamido)-3-phenylpropanamide (A4)

[0025] (S)-N-(4-methylbenzenesulfonyl)-2-(2-(5-((4'-methyl-[1,1'-biphenyl]-4-yl)methylene)-thiazolidine-2,4-dione-3-yl)acetamido)-3-phenylpropanamide (A5)

[0026] (S)-N-(4-chlorobenzenesulfonyl)-2-(2-(5-(([1,1'-biphenyl]-4-yl)methylene)-thiazolidine-2,4-dione-3-yl)acetamido)-3-phenylpropanamide (A6)

[0027] (S)-N-(3-nitro-4-chlorobenzenesulfonyl)-2-(2-(5-(([1,1'-biphenyl]-4-yl)methylene)-thiazolidine-2,4-dione-3-yl)acetamido)-3-phenylpropanamide (A7)

[0028] (S)-N-(4-nitrobenzenesulfonyl)-2-(2-(5-((4'-chloro-[1,1'-biphenyl]-4-yl)methylene)-thiazolidine-2,4-dione-3-yl)acetamido)-3-(4-bromophenyl)propanamide (A8)

[0029] (S)-N-(3-nitro-4-chlorobenzenesulfonyl)-2-(2-(5-((4'-chloro-[1,1'-biphenyl]-4-yl)methylene)-thiazolidine-2,4-dione-3-yl)acetamido)-3-(4-bromophenyl)propanamide (A9)

[0030] (S)-N-(4-nitrobenzenesulfonyl)-2-(2-(5-((4'-methyl-[1,1'-biphenyl]-4-yl)methylene)-thiazolidine-2,4-dione-3-yl)acetamido)-3-(4-bromophenyl)propanamide (A10)

[0031] (S)-N-(3-nitro-4-chlorobenzenesulfonyl)-2-(2-(5-(([1,1'-biphenyl]-4-yl)methylene)-thiazolidine-2,4-dione-3-yl)acetamido)-3-(4-bromophenyl)propanamide (A11)

[0032] (S)-N-(4-Nitrobenzenesulfonyl)-2-(2-(5-(([1,1'-biphenyl]-4-yl)methylene)-thiazolidine-2,4-dione-3-yl)acetamido)-3-(4-bromophenyl)propanamide (A12)

[0033] (S)-N-(4-Chlorobenzenesulfonyl)-2-(2-(5-(([1,1'-biphenyl]-4-yl)methylene)-thiazolidine-2,4-dione-3-yl)acetamido)-3-(4-bromophenyl)propanamide (A13)

[0034] (S)-N-(4-Methylbenzenesulfonyl)-2-(2-(5-(([1,1'-biphenyl]-4-yl)methylene)-thiazolidine-2,4-dione-3-yl)acetamido)-3-(4-bromophenyl)propanamide (A14)

[0035] (S)-N-(4-Nitrobenzenesulfonyl)-2-(2-(5-((4'-chloro-[1,1'-biphenyl]-4-yl)methylene)-thiazolidine-2,4-dione-3-yl)acetamido)-3-(4-(4-bromobenzyloxy)phenyl)propanamide (A15)

[0036] (S)-N-(4-Nitrobenzenesulfonyl)-2-(2-(5-((4'-methyl-[1,1'-biphenyl]-4-yl)methylene)-thiazolidine-2,4-dione-3-yl)acetamido)-3-(4-(4-bromobenzyloxy)phenyl)propanamide (A16)

[0037] (S)-N-(4-Nitrobenzenesulfonyl)-2-(2-(5-(([1,1'-biphenyl]-4-yl)methylene)-thiazolidine-2,4-dione-3-yl)acetamido)-3-(4-(4-bromobenzyloxy)phenyl)propanamide (A17)

[0038] (S)-N-(4-Nitrobenzenesulfonyl)-2-(2-(5-(([1,1'-biphenyl]-4-yl)methylene)-thiazolidine-2,4-dione-3-yl)acetamido)-3-(4-(benzyloxy)phenyl)propanamide (A18)

[0039] (S)-N-(3-Nitro-4-chlorobenzenesulfonyl)-2-(2-(5-(([1,1'-biphenyl]-4-yl)methylene)-thiazolidine-2,4-dione-3-yl)acetamido)-3-(4-(benzyloxy)phenyl)propanamide (A19)

[0040] (S)-N-(4-Nitrobenzenesulfonyl)-2-(2-(5-(([1,1'-biphenyl]-4-yl)methylene)-thiazolidine-2,4-dione-3-yl)acetamido)-3-(4-(4-nitrobenzyloxy)phenyl)propanamide (A20)

[0041] For the above preferred compounds, the numbers in the parentheses following are the numbers corresponding to the compound structures in the following reaction routes and Table 1. Detailed Description of the Invention

[0043] The terms and definitions used herein have the following meanings:

[0044] "Aryl" refers to an aromatic hydrocarbon containing a ring system, such as phenyl or naphthyl, which is optionally fused to a cycloalkyl, and the cycloalkyl preferably has 5-7 ring atoms, more preferably 5-6 ring atoms. Preferred aryl contains 5-15 carbon atoms.

[0045] "Heteroaryl" is an aromatic heterocycle, which can be a monocyclic or bicyclic group. They contain one or more, preferably 1-4, more preferably 1-3, even more preferably 1-2 heteroatoms, and the heteroatoms are independently selected from O, S, and N. Heteroaryl includes oxidized S or N, such as sulfinyl, sulfonyl, and N-oxide of tricyclic nitrogen. A carbon atom or a nitrogen atom is the connection point of the heteroaromatic ring structure, thereby maintaining a stable aromatic ring. Examples of heteroaryl include, but are not limited to, pyridyl, pyridazinyl, pyrazinyl, indolizinyl, benzothiophenyl, quinazolinyl, purinyl, indolyl, quinolinyl, pyrimidinyl, pyrrolyl, oxazolyl, thiazolyl, thiophenyl, isoxazolyl, oxadiazolyl, isothiazolyl, tetrazolyl, imidazolyl, triazinyl, furyl, benzofuryl, and indolyl.

[0046] "Arylalkyl" refers to an aryl linked by a C1-C6 alkylene.

[0047] "Heteroarylalkyl" refers to a heteroaryl linked by a C1-C6 alkylene.

[0048] "Arylalkenyl" refers to an aryl linked by a C2-C6 alkenyl.

[0049] "Heteroarylalkenyl" refers to a heteroaryl linked by a C2-C6 alkenyl.

[0050] "Alkyl", alone or in combination, refers to a group derived from an alkane and containing from 1 to 20 carbon atoms, preferably 1 to 12 carbon atoms (if not otherwise specified). It is a straight-chain or branched-chain alkyl group and includes a straight-chain or branched-chain alkyl group containing a cycloalkyl moiety or interrupted by a cycloalkyl moiety. The straight-chain or branched-chain alkyl group is connected at any available point to produce a stable compound. Examples thereof include, but are not limited to, 4-(isopropyl)-cyclohexylethyl or 2-methyl-cyclopropylpentyl. In many embodiments, the alkyl group is a straight-chain or branched-chain alkyl group containing 1 to 15 carbon atoms, 1 to 8 carbon atoms, 1 to 6 carbon atoms, 1 to 4 carbon atoms or 1 to 2 carbon atoms, such as methyl, ethyl, propyl, isopropyl, butyl, tert-butyl and similar alkyl groups.

[0051] "Alkylene" is a divalent alkane-derived carbon atom group that is straight-chain or branched-chain, in which two hydrogen atoms are removed from the same carbon atom or different carbon atoms. Examples of alkylene include, but are not limited to, -CH2-, -CH2CH2- and -CH2CH(CH3)-.

[0052] "Alkenyl", alone or in combination, as used herein refers to a straight-chain or branched-chain hydrocarbon containing 2-6, preferably 2-4 carbon atoms, and containing 1-2, preferably one carbon-carbon double bond. Examples of alkenyl include, but are not limited to, vinyl, propenyl, isopropenyl, butenyl.

[0053] "Cycloalkyl" is a substituted or unsubstituted, saturated or unsaturated cyclic group containing carbon atoms and / or one or more heteroatoms. The ring can be a monocyclic or fused-ring, bridged-ring or spiro-ring system. The number of ring atoms in each ring is 3-8, more preferably 3-6, such as cyclopropyl, cyclopentyl, cyclohexyl, adamantyl and similar groups.

[0054] "Alkoxy" represents the group -O-alkyl.

[0055] "Halogen", alone or in combination, refers to all halogens, namely chlorine (Cl), fluorine (F), bromine (Br) or iodine (I).

[0056] "Pharmaceutically acceptable salts" refers to salt forms of the compounds of formula (I) that are therapeutically effective and non-toxic. Anionic salts can be formed from any acidic group (such as a carboxyl group), or cationic salts can be formed from any basic group (such as an amino group). Many such salts are known in the art. Cationic salts are formed on any acidic group (such as a carboxyl group), or anionic salts are formed on any basic group (such as an amino group). Many of these salts are known in the art, such as cationic salts including salts of alkali metals (such as sodium and potassium) and alkaline earth metals (such as magnesium and calcium), as well as organic salts (such as ammonium salts). Anionic salts can also be conveniently obtained by treating the basic form of I with the corresponding acid, such acids including inorganic acids such as sulfuric acid, nitric acid, phosphoric acid, etc.; or organic acids such as acetic acid, propionic acid, glycolic acid, 2-hydroxypropionic acid, 2-oxopropionic acid, oxalic acid, malonic acid, succinic acid, maleic acid, fumaric acid, malic acid, tartaric acid, 2-hydroxy-1,2,3-propanetricarboxylic acid, methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, 4-methylbenzenesulfonic acid, cyclohexylsulfinic acid, 2-hydroxybenzoic acid, 4-amino-2-hydroxybenzoic acid, etc. These salts are well-known to those skilled in the art, and those skilled in the art can prepare any salt provided by the knowledge in the art. In addition, those skilled in the art can choose a certain salt over another based on factors such as solubility, stability, ease of formulation, etc. The determination and optimization of these salts are within the experience of those skilled in the art.

[0057] As used herein, "stereoisomers" defines all possible stereoisomeric forms of the compounds of the present invention or their physiological derivatives. Unless otherwise indicated, the chemical nomenclature of the compounds of the present invention includes mixtures of all possible stereochemical forms, said mixtures containing all diastereomers and enantiomers of the basic structural molecule, as well as individual isomeric forms of the compounds of the present invention that are substantially pure, i.e., containing less than 10%, preferably less than 5%, particularly less than 2%, and most preferably less than 1% of other isomers. All various stereoisomeric forms of the peptidomimetic compounds of the present invention are clearly included within the scope of the present invention.

[0058] The compounds of formula (I) can also exist in other protected forms or derivative forms, which are obvious to those skilled in the art and should all be included within the scope of the present invention.

[0059] The substituents as described above may themselves also be substituted by one or more substituents. Such substituents include those listed in C. Hansch and A. Leo, Substituent Constants for Correlation Analysis in Chemistry and Biology (1979). Preferred substituents include alkyl, alkenyl, alkoxy, hydroxy, oxo, nitro, amino, aminoalkyl (such as aminomethyl, etc.), cyano, halogen, carboxyl, carbonylalkoxy (such as carbonylethoxy, etc.), thio, aryl, cycloalkyl, heteroaryl, heterocycloalkyl (such as piperidinyl, morpholinyl, pyrrolyl, etc.), imino, hydroxyalkyl, aryloxy, arylalkyl and their combinations.

[0060] "Pharmaceutical composition" refers to a preparation containing a therapeutically significant amount of an active agent, which is prepared in a form suitable for administration to a patient. Thus, the preparation does not contain such an amount of any one or more components that a duly prudent medical practitioner would find the preparation unsuitable for administration to a normal subject. In many cases, such pharmaceutical composition is a sterile preparation.

[0061] Room temperature refers to the ambient temperature at which the experimental operation is carried out, which is controlled within the range of 10 - 30 °C.

[0062] II. Preparation method of substituted 3 - thiazolidine - 2,4 - dione Mcl - 1 protein inhibitors

[0063] The preparation method of substituted 3 - thiazolidine - 2,4 - dione Mcl - 1 protein inhibitors comprises the steps of: using thiazolidine - 2,4 - dione as the starting material, first carrying out Knoevenagel condensation reaction with differently substituted formaldehyde to generate intermediates 1a - 1c, then carrying out nucleophilic substitution reaction with methyl bromoacetate at the 3 - position N of thiazolidine and removing the methyl ester under acidic conditions to generate key intermediates 2a - 2c; in addition, carrying out amide condensation reaction with differently substituted amino acids 3a - 3e and differently substituted benzenesulfonamides to generate key intermediates 4a - 4k; finally, first removing the Boc protection of key intermediates 4a - 4k in ethyl acetate saturated with hydrogen chloride gas, and then using the mixed anhydride method, under the action of N - methylmorpholine and isobutyl chloroformate, carrying out amide condensation reaction with another key intermediate 2a - 2c to obtain target compounds A1 - A20.

[0064] The synthetic route is as follows:

[0065]

[0066] Wherein, the definitions of R1 - R3 are the same as those described in the above general formula (I);

[0067] Reagents and conditions: a) Different substituted formaldehydes, piperidine, ethanol, reflux; b) i. Methyl bromoacetate, potassium carbonate, acetone, reflux; ii. Glacial acetic acid, concentrated hydrochloric acid, reflux; c) 2-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU), N,N-diisopropylethylamine (DIEA), dichloromethane, ice bath - room temperature; d) i. Ethyl acetate solution saturated with hydrogen chloride, room temperature; ii. N-methylmorpholine, isobutyl chloroformate, tetrahydrofuran, -20 °C - room temperature.

[0068] The structural formula of the target compound in the synthesis route is shown in Table 1 below:

[0069]

[0070] Table 1 Structural formula of the target compound The specific operation steps of the said compound will be described in detail in the examples.

[0071]

[0072] Those skilled in the art can make changes to the above steps to improve the yield. They can determine the synthesis route based on the basic knowledge in this field, such as selecting reactants, solvents, and temperature, and can avoid side reactions by using various conventional protecting groups to improve the yield. These conventional protection methods can be found, for example, in T. Greene, Protecting Groups in Organic Synthesis.

[0073] III. Application of substituted 3-thiazolidine-2,4-dione Mcl-1 protein inhibitors

[0074] The present invention also provides the application of this series of compounds in the preparation of drugs for preventing or treating related mammalian diseases caused by abnormal expression of Mcl-1 protein. The mammalian diseases related to abnormal expression of Mcl-1 protein include cancer, neurodegenerative diseases, viral infections, inflammation, leukemia, malaria, and diabetes, etc.

[0075] In addition, the present invention also includes a pharmaceutical composition suitable for oral administration to mammals, comprising any compound of the general formula (I) above, and a pharmaceutically acceptable carrier, optionally comprising one or more pharmaceutically acceptable excipients.

[0076] In addition, the present invention also includes a pharmaceutical composition suitable for parenteral administration to mammals, comprising any compound of the general formula (I) and (II) above, and a pharmaceutically acceptable carrier, optionally comprising one or more pharmaceutically acceptable excipients.

[0077] Two aspects of enzyme activity inhibition and cell activity are tested to evaluate the in vitro biological activity of the compound.

[0078] In the in vitro enzyme inhibition experiment, the fluorescence polarization assay method was used. In the specific measurement system, the 5-FAM-labeled Bid-BH3 polypeptide was used as the fluorescent labeling molecule, which could specifically bind to the Mcl-1 protein. Its dissociation constant (K d ) was about 30 - 60 nM. After the two bound, a relatively high polarization value was generated. If the target compound to be tested could bind to the target protein, it would competitively inhibit the binding of Bid to this protein, resulting in a decrease in the polarization value. Then, the dose-effect curve of the competitive binding of the target compound was obtained, and finally the inhibition constant K i was calculated.

[0079] The MTT assay was used to test the cell activity of the compound. Tumor cell suspensions (myeloma cell line KM3, non-small cell lung cancer H1299) and normal liver cell line LO2 were respectively inoculated into 96-well plates. The culture medium containing different concentrations of the compound was added to each well. After incubation, MTT staining was performed. After continued incubation, the absorbance OD value of each well was measured at 570 nm on an enzyme-linked immunosorbent assay (ELISA) reader, and the cell growth inhibition rate was calculated to determine the activity of the compound.

[0080] The in vitro enzyme inhibition experiment showed that most of the compounds in the present invention had strong inhibitory activity against the Mcl-1 protein. Among them, the activities of compounds A1, A8, A9, etc. were comparable to those of the positive control drug UMI-77, and the inhibitory activity of compound A10 against the Mcl-1 protein was higher than that of the positive control drug UMI-77. At the same time, in the in vitro anti-tumor cell proliferation assay, compounds A1, A8, A9, and A10 had good inhibitory activity against the myeloma cell line KM3, which was comparable to that of the positive control drug AT-101, and had great development prospects and could be used to guide the discovery of new Mcl-1 protein inhibitors. Specific Embodiments

[0081] The present invention will be further described below in conjunction with the embodiments, but it is not limited thereto.

[0082] Example 1. Synthesis of 5-((4'-chloro-[1,1'-biphenyl]-4-yl)methylene)-thiazolidine-2,4-dione (1a)

[0083] Thiazolidine-2,4-dione (1.17 g, 10 mmol) was slowly added to anhydrous ethanol (80 mL). After dissolving with stirring at room temperature, piperidine (0.8 mL, 8 mmol) and 4'-chloro-biphenyl-4-carbaldehyde (2.27 g, 10.5 mmol) were added successively. Then the reaction was refluxed overnight in an oil bath. After cooling to room temperature, 200 mL of distilled water was first added, and then 2 mL of glacial acetic acid was slowly added dropwise. The mixture was stirred at room temperature for 30 min. The precipitate was collected by filtration, washed with water, dried in vacuo, and the resulting solid was recrystallized from methanol to obtain 2.23 g of a yellow crystalline solid 1a, with a yield of 71%, mp: 93 - 94 °C. 1 H NMR (500 MHz, DMSO-d6), δ 12.65 (s, 1H), 7.85 (d, J = 8.5 Hz, 3H), 7.77 (d, J = 8.5 Hz, 2H), 7.69 (d, J = 8.0 Hz, 2H), 7.55 (d, J = 8.5 Hz, 2H).

[0084] Synthesis of 2-(5-((4'-chloro-[1,1'-biphenyl]-4-yl)methylene)-thiazolidine-2,4-dione-3-yl)acetic acid (2a)

[0085] Compound 1a (2.21 g, 7 mmol) was dissolved in 100 mL of anhydrous acetone, and then potassium carbonate (1.93 g, 14 mmol) and methyl bromoacetate (1.3 mL, 14 mmol) were added. The reaction mixture was transferred to an oil bath and refluxed with stirring overnight. After cooling to room temperature, the filtrate was filtered and concentrated by rotary evaporation, and recrystallized from methanol to obtain 1.54 g of a yellow solid, with a yield of 59%. 1.24 g of it was dissolved in 18 mL of glacial acetic acid, and after dissolution, 4.5 mL of concentrated hydrochloric acid was added and the reaction mixture was transferred to an oil bath. After refluxing for 4 h, it was cooled overnight. The precipitate was collected by filtration, washed with water, dried in vacuo, to obtain 1.12 g of a yellow solid compound 2a, with a yield of 94%, mp: 100 - 101 °C. 1 H NMR (500 MHz, DMSO-d6), δ 13.43 (s, 1H), 8.04 (s, 1H), 7.89 (d, J = 8.5 Hz, 2H), 7.80 - 7.74 (m, 4H), 7.56 (d, J = 8.5 Hz, 2H), 4.41 (s, 2H).

[0086] (S)-tert-Butyl (1-(3-nitro-4-chlorobenzenesulfonamido)-1-oxo-3-phenylpropan-2-yl)carbamate (4a) synthesis

[0087] Under ice bath conditions, dissolve N-Boc-L-phenylalanine 3a (1.33 g, 5 mmol) in 30 mL of anhydrous dichloromethane, and then successively add DIEA (1.75 mL, 10 mmol) and HATU (2.29 g, 6 mmol). After stirring for 30 min, add 3-nitro-4-chlorobenzenesulfonamide (1.3 g, 1.1 mmol). Remove the ice bath and stir overnight at room temperature. Spin-dry the solvent. Then extract twice with ethyl acetate, combine the ethyl acetate layers, wash with saturated brine, and dry over anhydrous sodium sulfate. After filtration and spin-drying, purify by silica gel column chromatography (petroleum ether:ethyl acetate = 8:1 - 6:1) to obtain 1.96 g of white solid 4a with a yield of 81%, mp: 118 - 119 °C. 1 H NMR (500 MHz, CDCl3-d6), δ 10.22 (s, 1H), 8.45 (s, 1H), 8.17 - 8.25 (m, 1H), 7.72 (d, J = 8.5 Hz, 1H), 7.20 - 7.19 (m, 3H), 7.03 (d, J = 4.5 Hz, 2H), 5.16 (s, 1H), 4.38 (s, 1H), 3.04 - 3.00 (m, 1H), 2.93 - 2.89 (m, 1H), 1.38 (s, 9H).

[0088] (S)-N-(3-Nitro-4-chlorobenzenesulfonyl)-2-(2-(5-((4'-chloro-[1,1'-biphenyl]-4-yl)methylene)-thiazolidine-2,4-dione-3-yl)acetamido)-3-phenylpropanamide (LJB-II-20) (A1) Synthesis

[0089] Dissolve a key intermediate 4a (0.58 g, 1.2 mmol) in 20 mL of ethyl acetate solution saturated with hydrogen chloride gas, stir overnight at room temperature, filter and dry it, and collect this hydrochloride intermediate for later use. Dissolve another key intermediate 2a (0.37 g, 1 mmol) in 20 mL of anhydrous tetrahydrofuran and stir at -20 °C. Every 10 min, successively add N-methylmorpholine (0.24 mL, 2.2 mmol) and isobutyl chloroformate (0.2 mL, 1.5 mmol). Continue to stir for 1 h, add the reserved hydrochloride intermediate, slowly warm to room temperature, and stir overnight. Spin-dry the reaction solvent, then add an appropriate amount of ethyl acetate and transfer it to a separatory funnel. Wash twice with 1 mol / L hydrochloric acid solution and saturated brine respectively, dry over anhydrous sodium sulfate, filter and spin-dry, and then purify and separate by column chromatography (petroleum ether:ethyl acetate = 1:1 - 1:2) to obtain 0.11 g of light yellow solid A1, yield: 14%, mp: 208 - 209 °C. 11H NMR (500 MHz, DMSO-d6), δ 8.37 (s, 1H), 8.18 (s, 1H), 8.05 - 8.00 (m, 2H), 7.89 - 7.84 (m, 3H), 7.81 - 7.74 (m, 4H), 7.57 (d, J = 7.5 Hz, 2H), 7.12 (s, 3H), 6.99 (s, 2H), 4.32 - 4.27 (m, 3H), 2.97 (d, J = 8.5 Hz, 1H), 2.83 (d, J = 6.0 Hz, 1H). 13 13C NMR (125 MHz, DMSO-d6), δ 166.80, 165.18, 164.14, 146.47, 140.59, 137.56, 133.17, 132.62, 132.21, 131.52, 130.85, 129.30, 129.03, 128.58, 127.65, 127.42, 125.92, 124.48, 121.08, 56.17, 43.26, 37.61.

[0090] Example 2. Synthesis of (S)-N-(4-methylbenzenesulfonyl)-2-(2-(5-((4'-chloro-[1,1'-biphenyl]-4-yl)methylene)-thiazolidine-2,4-dione-3-yl)acetamido)-3-phenylpropanamide (A2)

[0091] The preparation methods of the intermediate and the target compound are as in Example 1. The yield is 20%, mp: 228 - 229 °C. 1 1H NMR (500 MHz, DMSO-d6), δ 12.44 (s, 1H), 8.66 (d, J = 7.0 Hz, 1H), 7.99 (s, 1H), 7.88 (d, J = 8.0 Hz, 2H), 7.79 - 7.73 (m, 6H), 7.56 (d, J = 8.0 Hz, 2H), 7.43 (d, J = 7.5 Hz, 2H), 7.21 - 7.11 (m, 5H), 4.52 (d, J = 5.5 Hz, 1H), 4.30 - 4.20 (m, 2H), 2.94 - 2.91 (m, 1H), 2.73 - 2.69 (m, 1H), 2.41 (s, 3H). 1313C NMR (125 MHz, DMSO-d6), δ 170.35, 167.22, 165.57, 165.53, 144.75, 141.15, 138.04, 136.78, 133.69, 133.24, 132.69, 131.36, 130.01, 129.66, 129.53, 129.07, 128.58, 128.04, 127.93, 127.02, 121.49, 54.94, 43.61, 37.15, 21.58.

[0092] Example 3. Synthesis of (S)-N-(4-chlorobenzenesulfonyl)-2-(2-(5-((4'-chloro-[1,1'-biphenyl]-4-yl)methylene)-thiazolidine-2,4-dione-3-yl)acetamido)-3-phenylpropanamide (A3)

[0093] The preparation methods of the intermediate and the target compound are as in Example 1. The yield is 26%, mp: 238 - 239 °C. 1 1H NMR (500 MHz, DMSO-d6), δ 12.63 (s, 1H), 8.70 (d, J = 7.5 Hz, 1H), 7.80 (s, 1H), 7.90 - 7.87 (m, 4H), 7.80 (d, J = 8.0 Hz, 2H), 7.75 - 7.69 (m, 4H), 7.57 (d, J = 8.5 Hz, 2H), 7.21 - 7.11 (m, 5H), 4.52 - 4.49 (m, 1H), 4.31 - 4.22 (m, 2H), 2.94 - 2.90 (m, 1H), 2.76 - 2.72 (m, 1H). 13 13C NMR (125 MHz, DMSO-d6), δ 170.59, 167.24, 165.61, 165.58, 141.15, 139.20, 138.25, 138.04, 136.68, 133.70, 133.27, 132.69, 131.37, 130.00, 129.75, 129.65, 129.53, 129.07, 128.58, 127.92, 127.06, 121.48, 55.03, 43.60, 37.02.

[0094] Example 4. Synthesis of (S)-N-(3-nitro-4-chlorobenzenesulfonyl)-2-(2-(5-((4'-methyl-[1,1'-biphenyl]-4-yl)methylene)-thiazolidine-2,4-dione-3-yl)acetamido)-3-phenylpropanamide (A4)

[0095] The preparation methods of the intermediate and the target compound are as in Example 1. The yield is 25%, mp: 195 - 196 °C.1 1H NMR (500 MHz, DMSO-d6), δ 8.40 (s, 1H), 8.33 (s, 1H), 8.07 (d, J = 7.5 Hz, 1H), 7.99 (s, 1H), 7.92 (d, J = 8.0 Hz, 1H)), 7.86 (d, J = 8.0 Hz, 2H), 7.73 (d, J = 8.6 Hz, 2H), 7.67 (d, J = 8.0 Hz, 2H)), 7.32 (d, J = 7.5 Hz, 2H), 7.14 - 7.03 (m, 5H), 4.31 - 4.23 (m, 3H), 2.97 - 2.93 (m, 1H), 2.82 - 2.78 (m, 1H), 2.36 (s, 3H). 13 13C NMR (125 MHz, DMSO-d6), δ 170.81, 170.21, 167.25, 165.58, 165.55, 144.61, 141.16, 139.77, 138.05, 136.27, 133.69, 133.24, 132.70, 131.90, 131.39, 129.95, 129.54, 129.10, 128.04, 127.95, 121.50, 120.29, 54.74, 43.65, 36.53, 21.59.

[0096] Example 5. Synthesis of (S)-N-(4-methylbenzenesulfonyl)-2-(2-(5-((4'-methyl-[1,1'-biphenyl]-4-yl)methylene)-thiazolidine-2,4-dione-3-yl)acetamido)-3-phenylpropanamide (A5)

[0097] The preparation methods of the intermediate and the target compound are as in Example 1. The yield is 45%, mp: 251 - 252 °C. 1 1H NMR (500 MHz, DMSO-d6), δ 12.46 (s, 1H), 8.66 (s, 1H), 7.99 (s, 1H), 7.85 - 7.66 (m, 8H)), 7.43 - 7.11 (m, 9H), 4.51 (s, 1H), 4.26 (d, J = 11.5 Hz, 2H), 2.93 - 2.91 (m, 1H), 2.73 - 2.70 (m, 1H), 2.41 (s, 3H), 2.36 (s, 3H). 1313C NMR (125 MHz, DMSO-d6), δ 170.35, 167.29, 165.61, 165.56, 144.77, 142.53, 138.33, 136.78, 136.67, 136.32, 133.45, 132.04, 131.36, 130.19, 130.02, 129.67, 128.59, 128.05, 127.66, 127.14, 127.04, 120.97, 54.93, 43.60, 37.14, 21.58, 21.21.

[0098] Example 6. Synthesis of (S)-N-(4-chlorobenzenesulfonyl)-2-(2-(5-(([1,1'-biphenyl]-4-yl)methylene)-thiazolidine-2,4-dione-3-yl)acetamido)-3-phenylpropanamide (A6)

[0099] The preparation methods of the intermediate and the target compound are as in Example 1. The yield is 26%, mp: 222 - 223 °C. 1 1H NMR (500 MHz, DMSO-d6), δ 12.61 (s, 1H), 8.50 (d, J = 5.5 Hz, 1H), 7.93 (s, 1H), 7.81 - 7.80 (m, 4H), 7.70 - 7.66 (m, 4H), 7.60 (d, J = 8.5 Hz, 2H), 7.45 - 7.42 (m, 2H), 7.37 - 7.34 (m, 1H), 7.12 - 7.01 (m, 5H), 4.41 - 4.37 (m, 1H), 4.24 - 4.15 (m, 2H), 2.88 - 2.84 (m, 1H), 2.71 - 2.66 (m, 1H). 13 13C NMR (125 MHz, DMSO-d6), δ 171.22, 167.28, 165.62, 165.41, 142.57, 139.26, 136.98, 133.37, 132.38, 131.34, 129.91, 129.68, 129.57, 129.47, 128.77, 128.50, 127.97, 127.31, 126.93, 121.26, 55.32, 43.65, 37.23.

[0100] Example 7. Synthesis of (S)-N-(3-nitro-4-chlorobenzenesulfonyl)-2-(2-(5-(([1,1'-biphenyl]-4-yl)methylene)-thiazolidine-2,4-dione-3-yl)acetamido)-3-phenylpropanamide (A7)

[0101] The preparation methods of the intermediate and the target compound are as described in Example 1. The yield is 22%, mp: 232 - 233 °C. 1H NMR (500 MHz, DMSO-d6), δ 8.70 (d, J = 6.5 Hz, 1H), 8.49 (d, J = 2.5 Hz, 1H), 8.12 (dd, J1 = 2.0 Hz, J2 = 8.5 Hz, 1H), 8.02 - 8.00 (m, 2H), 7.89 (d, J = 8.5 Hz, 2H), 7.77 - 7.74 (m, 4H), 7.52 (t, J = 7.5 Hz, 2H), 7.44 - 7.41 (m, 1H), 7.20 - 7.19 (m, 3H), 7.12 - 7.11 (m, 2H), 4.51 - 4.46 (m, 1H), 4.30 - 4.23 (m, 2H), 2.96 - 2.92 (m, 1H), 2.82 - 2.77 (m, 1H). 13 13C NMR (125 MHz, DMSO-d6), δ 171.22, 167.29, 165.67, 165.62, 147.51, 142.58, 139.54, 139.26, 136.65, 133.40, 132.87, 132.38, 131.37, 131.14, 129.68, 129.59, 128.79, 128.54, 128.00, 127.33, 127.05, 125.61, 121.23, 55.24, 43.57, 36.94.

[0102] Example 8. Synthesis of (S)-N-(4-Nitrobenzenesulfonyl)-2-(2-(5-((4'-Chloro-[1,1'-biphenyl]-4-yl)methylene)-thiazolidine-2,4-dione-3-yl)acetamido)-3-(4-bromophenyl)propanamide (A8)

[0103] The preparation methods of the intermediate and the target compound are as described in Example 1. The yield is 36%, mp: 198 - 199 °C. 1 1H NMR (500 MHz, DMSO-d6), δ 8.49 (s, 1H), 8.36 (d, J = 8.5 Hz, 2H), 8.08 (d, J = 9.0 Hz, 2H), 8.00 (s, 1H), 7.89 (d, J = 8.0 Hz, 2H), 7.80 (d, J = 8.5 Hz, 2H), 7.75 (d, J = 8.0 Hz, 2H), 7.57 (d, J = 8.5 Hz, 2H), 7.35 (d, J = 8.0 Hz, 2H), 7.06 (d, J = 8.0 Hz, 2H), 4.38 (d, J = 6.0 Hz, 1H), 4.26 (s, 2H), 2.95 - 2.91 (m, 1H), 2.79 - 2.75 (m, 1H). 1313C NMR (125 MHz, DMSO-d6), δ 167.27, 165.61, 165.23, 149.80, 141.13, 138.03, 136.84, 133.68, 133.21, 132.69, 131.96, 131.36, 131.26, 129.52, 129.38, 129.07, 127.91, 124.26, 121.49, 120.06, 55.66, 43.71, 36.85.

[0104] Example 9. Synthesis of (S)-N-(3-nitro-4-chlorobenzenesulfonyl)-2-(2-(5-((4'-chloro-[1,1'-biphenyl]-4-yl)methylene)-thiazolidine-2,4-dione-3-yl)acetamido)-3-(4-bromophenyl)propanamide (A9)

[0105] The preparation methods of the intermediate and the target compound are as in Example 1. The yield is 37%, mp: 232 - 233 °C. 1 1H NMR (500 MHz, DMSO-d6), δ 8.44 (s, 1H), 8.09 (d, J = 8.5 Hz, 1H), 8.00 (s, 1H), 7.94 - 7.88 (m, 4H), 7.80 (d, J = 8.5 Hz, 2H), 7.76 (d, J = 8.5 Hz, 2H), 7.57 (d, J = 8.5 Hz, 2H), 7.33 (d, J = 8.5 Hz, 2H), 7.01 (d, J = 8.0 Hz, 2H), 4.35 - 4.34 (m, 1H), 4.27 (s, 2H), 2.95 - 2.91 (m, 1H), 2.80 - 2.76 (m, 1H). 13 13C NMR (125 MHz, DMSO-d6), δ 166.72, 165.07, 164.77, 146.74, 140.60, 140.56, 137.51, 136.17, 133.15, 133.07, 132.67, 132.64, 132.26, 132.17, 131.43, 130.82, 130.68, 128.97, 128.51, 127.36, 124.83, 120.96, 119.64, 55.08, 43.20, 36.25.

[0106] Example 10. Synthesis of (S)-N-(4-nitrobenzenesulfonyl)-2-(2-(5-((4'-methyl-[1,1'-biphenyl]-4-yl)methylene)-thiazolidine-2,4-dione-3-yl)acetamido)-3-(4-bromophenyl)propanamide (A10)

[0107] The preparation methods of the intermediate and the target compound are as described in Example 1. The yield is 39%, mp: 211 - 212 °C. 1 H NMR (500 MHz, DMSO-d6), δ 12.87 (s, 1H), 8.72 (d, J = 7.0 Hz, 1H), 8.42 (d, J = 8.5 Hz, 2H), 8.13 (d, J = 8.5 Hz, 2H), 7.98 (s, 1H), 7.86 (d, J = 8.0 Hz, 2H), 7.73 (d, J = 8.0 Hz, 2H), 7.67 (d, J = 8.0 Hz, 2H), 7.40 (d, J = 8.0 Hz, 2H), 7.32 (d, J = 8.0 Hz, 2H), 7.11 (d, J = 8.0 Hz, 2H), 4.50 (dd, J1 = 7.5 Hz, J2 = 14.0 Hz, 1H), 4.25 (s, 2H), 2.93 - 2.90 (m, 1H), 2.77 - 2.73 (m, 1H), 2.36 (s, 3H). 13 C NMR (125 MHz, DMSO-d6), δ 170.93, 167.29, 165.69, 165.60, 150.59, 142.53, 138.32, 136.31, 136.22, 133.47, 132.01, 131.90, 131.45, 131.35, 130.18, 129.68, 127.63, 127.12, 124.78, 120.91, 120.36, 55.00, 43.57, 36.30, 21.21.

[0108] Example 11. Synthesis of (S)-N-(3-nitro-4-chlorobenzenesulfonyl)-2-(2-(5-(([1,1'-biphenyl]-4-yl)methylene)-thiazolidine-2,4-dione-3-yl)acetamido)-3-(4-bromophenyl)propanamide (A11)

[0109] The preparation methods of the intermediate and the target compound are as described in Example 1. The yield is 17%, mp: 178 - 179 °C. 1 H NMR (500 MHz, DMSO-d6), δ 8.40 (s, 1H), 8.24 (s, 1H), 8.06 - 8.01 (m, 2H), 7.94 - 7.87 (m, 3H), 7.77 - 7.74 (m, 4H), 7.52 - 7.50 (m, 2H), 7.44 - 7.41 (m, 1H), 7.30 (d, J = 7.0 Hz, 2H), 6.97 (d, J = 7.0 Hz, 2H), 4.28 (s, 3H), 2.95 - 2.92 (m, 1H), 2.81 - 2.77 (m, 1H). 1313C NMR (125 MHz, DMSO-d6), δ 173.59, 166.97, 165.17, 164.63, 146.51, 143.42, 142.02, 138.59, 136.48, 132.86, 132.17, 131.84, 131.69, 131.42, 130.82, 130.56, 129.09, 128.28, 128.11, 127.37, 126.71, 124.48, 120.64, 119.41, 55.68, 43.16, 36.64.

[0110] Example 12. Synthesis of (S)-N-(4-Nitrobenzenesulfonyl)-2-(2-(5-(([1,1'-Biphenyl]-4-yl)methylene)-thiazolidine-2,4-dione-3-yl)acetamido)-3-(4-bromophenyl)propanamide (A12)

[0111] The preparation methods of the intermediate and the target compound are as in Example 1. The yield is 22%, mp: 184 - 185 °C. 1 1H NMR (500 MHz, DMSO-d6), δ 12.79 (s, 1H), 8.73 (d, J = 7.5 Hz, 1H), 8.42 (d, J = 9.0 Hz, 2H), 8.13 (d, J = 8.5 Hz, 2H), 7.99 (s, 1H), 7.89 (d, J = 8.5 Hz, 2H), 7.77 - 7.74 (m, 4H), 7.53 (t, J = 7.5 Hz, 2H), 7.44 - 7.37 (m, 3H), 7.11 (d, J = 8.5 Hz, 2H), 4.50 - 4.46 (m, 1H), 4.25 (s, 2H), 2.93 - 2.90 (m, 1H), 2.77 - 2.73 (m, 1H). 13 13C NMR (125 MHz, DMSO-d6), δ 170.80, 167.27, 165.71, 165.59, 150.68, 142.61, 139.26, 136.18, 133.41, 132.36, 131.89, 131.46, 131.35, 129.69, 129.58, 128.77, 127.98, 127.32, 124.81, 121.22, 120.37, 54.95, 43.59, 36.27.

[0112] Example 13. Synthesis of (S)-N-(4-Chlorobenzenesulfonyl)-2-(2-(5-(([1,1'-Biphenyl]-4-yl)methylene)-thiazolidine-2,4-dione-3-yl)acetamido)-3-(4-bromophenyl)propanamide (A13)

[0113] The preparation methods of the intermediate and the target compound are as described in Example 1. The yield is 55%, mp: 174 - 175 °C. 1 H NMR (500 MHz, DMSO-d6), δ 12.62 (s, 1H), 8.70 (d, J = 7.5 Hz, 1H), 8.01 (s, 1H), 7.89 - 7.84 (m, 4H), 7.77 - 7.74 (m, 4H), 7.71 (d, J = 8.5 Hz, 2H) 7.53 (t, J = 7.5 Hz, 2H), 7.44 - 7.38 (m, 3H), 7.08 (d, J = 8.5 Hz, 2H), 4.51 (dd, J1 = 8.0, J2 = 13.5 Hz, 1H), 4.29 - 4.22 (m, 2H), 2.91 - 2.87 (m, 1H), 2.74 - 2.69 (m, 1H). 13 C NMR (125 MHz, DMSO-d6), δ 169.81, 166.80, 165.20, 165.10, 142.08, 138.80, 138.74, 137.60, 135.61, 132.91, 131.85, 131.38, 130.93, 130.87, 129.51, 129.27, 129.09, 128.28, 127.48, 126.82, 120.72, 119.89, 54.24, 43.10, 36.20.

[0114] Example 14. Synthesis of (S)-N-(4-methylbenzenesulfonyl)-2-(2-(5-(([1,1'-biphenyl]-4-yl)methylene)-thiazolidine-2,4-dione-3-yl)acetamido)-3-(4-bromophenyl)propanamide (A14)

[0115] The preparation methods of the intermediate and the target compound are as described in Example 1. The yield is 18%, mp: 182 - 183 °C. 1 H NMR (500 MHz, DMSO-d6), δ 12.43 (s, 1H), 8.68 (d, J = 6.5 Hz, 1H), 8.00 (s, 1H), 7.89 (d, J = 7.0 Hz, 2H), 7.77 - 7.75 (m, 6H), 7.51 - 7 - 37 (m, 7H), 7.07 (d, J = 7.0 Hz, 2H), 4.51 (s, 1H), 4.29 - 4.22 (m, 2H), 2.90 - 2.88 (m, 1H), 2.71 - 2.67 (m, 1H), 2.42 (s, 3H). 1313C NMR (125 MHz, DMSO-d6), δ 169.61, 166.79, 165.10, 144.26, 142.08, 138.74, 136.16, 135.71, 132.88, 131.85, 131.38, 130.91, 130.86, 129.49, 129.08, 128.28, 127.55, 127.48, 126.82, 120.73, 119.82, 54.18, 43.13, 36.00, 21.09.

[0116] Example 15. Synthesis of (S)-N-(4-Nitrobenzenesulfonyl)-2-(2-(5-((4'-Chloro-[1,1'-biphenyl]-4-yl)methylene)-thiazolidine-2,4-dione-3-yl)acetamido)-3-(4-(4-Bromobenzyloxy)phenyl)propanamide (A15)

[0117] The preparation methods of the intermediates and the target compound are as in Example 1. The yield is 15%, mp: 189 - 190 °C. 1 1H NMR (500 MHz, DMSO-d6), δ 8.67 (d, J = 6.5 Hz, 1H), 8.41 (d, J = 8.5 Hz, 1H), 8.14 - 8.10 (m, 1H), 8.00 - 7.97 (m, 1H), 7.89 - 7.85 (m, 2H), 7.80 - 7.73 (m, 4H), 7.60 - 7.55 (m, 4H), 7.41 - 7.37 (m, 2H), 7.16 (d, J = 8.0 Hz, 1H), 7.06 (d, J = 8.0 Hz, 2H), 6.94 - 6.81 (m, 3H), 5.06 - 5.01 (m, 2H), 4.44 (d, J = 7.0 Hz, 1H), 4.25 (s, 2H), 2.88 - 2.84 (m, 1H), 2.73 - 2.68 (m, 1H). 13 13C NMR (125 MHz, DMSO-d6), δ 166.74, 165.09, 164.91, 156.79, 140.64, 140.60, 137.53, 136.60, 136.56, 133.18, 132.76, 132.73, 132.17, 131.29, 130.86, 130.25, 129.66, 129.03, 128.57, 127.41, 124.06, 120.94, 120.84, 114.34, 68.24, 55.18, 43.13, 35.85.

[0118] Synthesis of Example 16. (S)-N-(4-Nitrobenzenesulfonyl)-2-(2-(5-((4'-Methyl-[1,1'-biphenyl]-4-yl)methylene)-thiazolidine-2,4-dione-3-yl)acetamido)-3-(4-(4-Bromobenzyloxy)phenyl)propanamide (A16)

[0119] The preparation methods of the intermediate and the target compound are as in Example 1. The yield is 36%, mp: 183 - 184 °C. 1 H NMR (400 MHz, DMSO-d6), δ 8.68 (d, J = 7.2 Hz, 1H), 8.39 (d, J = 8.8 Hz, 2H), 8.10 (d, J = 8.8 Hz, 2H), 7.95 (s, 1H), 7.84 (d, J = 8.0 Hz, 2H), 7.70 (d, J = 8.0 Hz, 2H), 7.65 (d, J = 8.0 Hz, 2H), 7.58 (d, J = 8.4 Hz, 2H), 7.39 (d, J = 8.4 Hz, 2H), 7.30 (d, J = 8.0 Hz, 2H), 7.04 (d, J = 8.4 Hz, 2H), 6.83 (d, J = 8.0 Hz, 2H), 5.02 (s, 2H), 4.44 - 4.38 (m, 1H), 4.22 (s, 2H), 2.86 - 2.81 (m, 1H), 2.70 - 2.64 (m, 1H), 2.33 (s, 3H). 13 C NMR (100 MHz, DMSO-d6), δ 170.58, 166.79, 165.16, 165.11, 156.87, 150.15, 148.34, 142.03, 137.83, 136.59, 135.81, 132.97, 131.51, 131.30, 130.86, 130.23, 129.69, 129.19, 128.42, 127.15, 126.64, 124.31, 120.85, 120.42, 114.42, 68.25, 54.87, 43.05, 35.60, 20.71.

[0120] Synthesis of Example 17. (S)-N-(4-Nitrobenzenesulfonyl)-2-(2-(5-(([1,1'-Biphenyl]-4-yl)methylene)-thiazolidine-2,4-dione-3-yl)acetamido)-3-(4-(4-Bromobenzyloxy)phenyl)propanamide (A17)

[0121] The preparation methods of the intermediate and the target compound are as in Example 1. The yield is 31%, mp: 155 - 156 °C. 11H NMR (400 MHz, DMSO-d6), δ 12.86 (s, 1H), 8.52 (s, 1H), 8.35 (d, J = 8.8 Hz, 2H), 8.06 (d, J = 8.4 Hz, 2H), 7.96 (s, 1H), 7.85 (d, J = 8.0 Hz, 2H), 7.74 - 7.69 (m, 4H), 7.57 (d, J = 8.4 Hz, 2H), 7.49 (t, J = 7.6 Hz, 2H), 7.41 - 7.36 (m, 3H), 7.00 (d, J = 8.4 Hz, 2H), 6.79 (d, J = 8.4 Hz, 2H), 5.00 (s, 2H), 4.34 - 4.31 (m, 1H), 4.22 (s, 2H), 2.86 - 2.81 (m, 1H), 2.70 - 2.64 (m, 1H). 13 13C NMR (100 MHz, DMSO-d6), δ 171.06, 167.23, 165.66, 165.58, 157.16, 150.68, 147.43, 145.61, 141.18, 138.04, 133.70, 133.27, 132.67, 131.38, 130.81, 129.70, 129.54, 129.22, 129.09, 128.61, 127.94, 124.84, 124.07, 121.44, 114.97, 68.41, 55.36, 43.57, 36.10.

[0122] Example 18. Synthesis of (S)-N-(4-Nitrobenzenesulfonyl)-2-(2-(5-(([1,1'-Biphenyl]-4-yl)methylene)-thiazolidine-2,4-dione-3-yl)acetamido)-3-(4-(Benzyloxy)phenyl)propanamide (A18)

[0123] The preparation methods of the intermediate and the target compound are as in Example 1. The yield is 34%, mp: 188 - 189 °C. 1 1H NMR (400 MHz, DMSO-d6), δ 12.86 (s, 1H), 8.72 (s, 1H), 8.42 (d, J = 7.2 Hz, 2H), 8.13 (d, J = 7.2 Hz, 2H), 8.00 (s, 1H), 7.89 - 7.75 (m, 6H), 7.51 - 7.34 (m, 8H), 7.07 (d, J = 6.4 Hz, 2H), 6.86 (d, J = 6.0 Hz, 2H), 5.06 (s, 2H), 4.45 (s, 1H), 4.26 (s, 2H), 2.87 - 2.85 (m, 1H), 2.73 - 2.68 (m, 1H). 13CNMR(125MHz, DMSO-d6), δ 167.26, 165.63, 165.24, 157.01, 147.41, 147.28, 145.66, 141.13, 138.04, 133.69, 133.20, 132.81, 132.71, 131.36, 130.86, 129.75, 129.53, 129.08, 128.57, 127.93, 125.40, 124.07, 121.52, 114.75, 68.39, 55.97, 43.71, 36.57.

[0124] Example 19. Synthesis of (S)-N-(3-Nitro-4-chlorobenzenesulfonyl)-2-(2-(5-(([1,1'-Biphenyl]-4-yl)methylene)-thiazolidine-2,4-dione-3-yl)acetamido)-3-(4-(benzyloxy)phenyl)propanamide (A19)

[0125] The preparation methods of the intermediates and the target compound are as in Example 1. The yield is 24%, mp: 216 - 217 °C. 1 H NMR(400MHz, DMSO-d6), δ 12.86(s, 1H), 8.73(d, J = 6.8Hz, 1H), 8.53(d, J = 2.0Hz, 1H), 8.14 - 8.12(m, 1H), 8.03 - 8.00(m, 2H), 7.89(d, J = 8.4Hz, 2H), 7.77 - 7.73(m, 4H), 7.53 - 7.34(m, 8H), 7.05(d, J = 8.4Hz, 2H), 6.86(d, J = 8.4Hz, 2H), 5.06(s, 2H), 4.48(dd, J1 = 7.2Hz, J2 = 13.6Hz, 1H), 4.28(s, 2H), 2.90 - 2.86(m, 1H), 2.75 - 2.70(m, 1H). 13 C NMR(100MHz, DMSO-d6), δ 171.10, 167.29, 165.71, 165.62, 157.64, 147.53, 142.59, 139.26, 137.60, 133.46, 133.40, 132.87, 132.38, 131.36, 131.32, 130.74, 129.59, 128.90, 128.78, 128.66, 128.27, 128.12, 127.99, 127.32, 125.69, 121.25, 114.84, 69.61, 55.40, 43.58, 36.10.

[0126] Synthesis of Example 20. (S)-N-(4-Nitrobenzenesulfonyl)-2-(2-(5-(([1,1'-Biphenyl]-4-yl)methylene)-thiazolidine-2,4-dione-3-yl)acetamido)-3-(4-(4-Nitrobenzyloxy)phenyl)propanamide (A20)

[0127] The preparation methods of the intermediate and the target compound are as in Example 1. The yield is 50%, mp: 205 - 206 °C. 1 H NMR (400 MHz, DMSO-d6), δ 12.88 (s, 1H), 8.47 (s, 1H), 8.37 (d, J = 8.8 Hz, 2H), 8.28 (d, J = 8.8 Hz, 2H), 8.09 (d, J = 8.8 Hz, 2H), 7.99 (s, 1H), 7.89 (d, J = 8.4 Hz, 2H), 7.77 - 7.70 (m, 6H), 7.53 - 7.49 (m, 2H), 7.45 - 7.41 (m, 1H), 7.04 (d, J = 8.4 Hz, 2H), 6.85 (d, J = 8.4 Hz, 2H), 5.23 (s, 2H), 4.36 - 4.21 (m, 3H), 2.92 - 2.87 (m, 1H), 2.75 - 2.70 (m, 1H). 13 C NMR (100 MHz, DMSO-d6), δ 167.30, 165.65, 165.20, 156.98, 149.83, 147.41, 145.67, 142.56, 139.24, 133.34, 132.36, 131.34, 130.85, 129.92, 129.58, 129.41, 128.78, 128.57, 127.96, 127.31, 124.31, 124.06, 121.23, 114.82, 68.39, 56.03, 43.70, 36.62.

[0128] Activity evaluation of the target compound

[0129] Experimental Example 1. Inhibition test of the target compound on Mcl-1 protein (In vitro)

[0130] Experimental reagents:

[0131] Bid-BH3 polypeptide fluorescently labeled with 5-FAM at the N-terminus

[0132] (5-FAM-QEDIIRNIARHLAQVGDSMDRSIPPG), dissolved in 1×PBS;

[0133] Test buffer: 1×PBS;

[0134] Calibration solution: 1 nM fluorescein, 10 mM NaOH;

[0135] Experimental instruments:

[0136] TECAN Genios Pro multi-functional microplate reader.

[0137] Experimental procedures:

[0138] (1) Add the target protein and the small molecule compound to be tested into the test buffer, mix well and incubate in the dark at room temperature for 30 min. Then add the fluorescently labeled Bid BH3 polypeptide to make the total volume of each solution 200 μL, mix well and incubate in the dark at room temperature for 20 min.

[0139] (2) Transfer 60 μL of each of the above solutions and the calibration solution to a black 384-well plate (in triplicate), and immediately perform fluorescence polarization detection on the microplate reader. Using 485 nm as the excitation wavelength and 535 nm as the emission wavelength, set the fluorescence polarization value of the calibration solution to 20 mP.

[0140] (3) All compounds are first screened at three typical concentrations (1 μM, 10 μM, 50 μM). Each compound is measured in triplicate on the same plate, and the average value of the polarization value measurements is taken. Calculate the inhibition rate based on the measurement results of the negative control, positive control, and the test compound. The target protein concentration usually used in the measurement is 300 - 500 nM, the fluorescently labeled polypeptide is 5-FAM-Bid-BH3 polypeptide, and the positive compound is AT-101. If the test results show that the inhibition rate of the compound is greater than 50% at a concentration of 50 μM and its inhibition rate shows an obvious dose-dependent relationship at the three tested concentrations, it is considered that the compound has specific binding to the target protein and further accurate IC 50 value needs to be measured.

[0141] (4) For the compounds showing obvious activity in the primary screening, measure the complete binding curve at 7 different concentrations (1 nM, 10 nM, 100 nM, 1 μM, 10 μM, 50 μM, 100 μM). Each compound is measured in triplicate on the same plate, and the average value of the polarization value measurements is taken. Use GraphPad Prism software to process the data and plot the graph to obtain the IC 50 value of the compound.

[0142] (5) Based on the total protein concentration, total fluorescent polypeptide concentration, dissociation constant of the protein-polypeptide complex, and the IC 50 value of the test compound used in the measurement, use the calculation method in the following literature to obtain the competitive inhibition constant K i(Nikolovska-Coleska, Z.; et al. Development and optimization of a binding assay for the XIAP BIR3 domain using fluorescence polarization. Anal Biochem. 2004, 332, 261-273).

[0143] The experimental results are shown in Table 2.

[0144] Table 2. Results of in vitro inhibition experiment of target compounds on Mcl-1 protein

[0145]

[0146] a The values in the table are the averages of the results of three trials. As can be seen from the table, most compounds showed inhibitory activity against Mcl-1 at the submicromolar level. When the amino acid was phenylalanine or 4-bromophenylalanine, and the sulfonamide moiety was 4-chloro-3-nitrobenzenesulfonamide or 4-nitrobenzenesulfonamide, the compounds showed good inhibitory activity against Mcl-1 protein, such as compounds A1, A4, A8, A9, A10, and A11. Among them, the inhibitory activities of compounds A1, A4, A8, A9, and A11 against Mcl-1 were comparable to those of the positive drug UMI-77. In addition, the compound A10 with the highest activity showed inhibitory activity higher than that of the positive control drug, which is of great significance for the further development of more active Mcl-1 protein inhibitors and the preparation of drugs for preventing and treating related mammalian diseases caused by abnormal expression of Mcl-1 protein.

[0147] Experimental Example 2. Activity test of target compounds for inhibiting cell proliferation (In vitro)

[0148] Four compounds with better enzyme activities were selected for the activity test of inhibiting cancer cell proliferation in vitro, and the results are shown in Table 3.

[0149] Term description:

[0150] Myeloma cell line KM3, non-small cell lung cancer H1299, and normal hepatocyte cell line LO2.

[0151] IC 50 : Half maximal inhibitory concentration.

[0152] Materials:

[0153] KM3, H1299, LO2, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), 10% fetal bovine serum, 96-well plates

[0154] Methods:

[0155] The cell lines used in cell culture were cultured conventionally. Cells in the logarithmic growth phase were used in all experiments.

[0156] Cell growth detection (MTT method): The cell suspension was adjusted to 5×10 4 / mL (suspension cells were adjusted to 10 5 / mL), and were inoculated into 96-well plates (100 μL / well), with 2000 - 5000 cells / well. After plating for 4 h, 100 μL of medium containing compounds at different concentrations was added to each well. Three replicate wells were set for each concentration. The wells without cells were used as blanks when reading, and the wells with cells but without compounds were used as compound blanks. UMI-77 was used as a compound positive control. They were incubated at 37 °C in 5% CO2 for 48 h. Then 10 μL of 0.5% MTT staining solution was added to each well and incubation continued. After 4 h, centrifugation was carried out at 2500 rpm for 30 min, and then the medium in the plate wells was discarded. 150 μL of DMSO was added to each well, and they were shaken at a constant temperature of 37 °C for 5 - 10 min. The absorbance OD value of each well was measured at 570 nm on an enzyme-linked immunosorbent assay (ELISA) reader. The cell growth inhibition rate was calculated according to the following formula:

[0157]

[0158] Table 3 Anti-tumor cell proliferation experimental results of compounds A1, A8, A9, and A10

[0159]

[0160] a The values in the table are the averages of three tests, and the values after "±" represent the standard deviation

[0161] In vitro anti-proliferation activity experiments of two tumor cell lines, KM3 and H1299, and normal liver cell line LO2 were carried out on 4 compounds with good enzyme activity. The test data showed that the 4 target compounds tested had inhibitory effects on both tumor cell lines, especially on KM3 cells. At the same time, these 4 compounds had almost no effect on the growth of normal cells. And their inhibitory activity on KM3 cells was comparable to that of the positive control UMI-77, indicating that substituted 3-thiazolidine-2,4-dione Mcl-1 protein inhibitors have good development prospects and can be further studied for their activity to develop more active compounds for the preparation of drugs for preventing and treating related mammalian diseases caused by abnormal Mcl-1 protein expression.

Claims

1. A substituted 3-thiazolidine-2,4-dione Mcl-1 protein inhibitor, which is a compound with the structure of general formula (I) or a pharmaceutically acceptable salt thereof: The general formula (I) is one of the following compounds: (S)-N-(3-Nitro-4-chlorobenzenesulfonyl)-2-(2-(5-((4'-chloro-[1,1'-biphenyl]-4-yl)methylene)-thiazolidine-2,4-dione-3-yl)acetamido)-3-phenylpropanamide (LJB-II-20) (A1); (S)-N-(3-Nitro-4-chlorobenzenesulfonyl)-2-(2-(5-((4'-methyl-[1,1'-biphenyl]-4-yl)methylene)-thiazolidine-2,4-dione-3-yl)acetamido)-3-phenylpropanamide (LJB-III-10) (A4); (S)-N-(4-Nitrobenzenesulfonyl)-2-(2-(5-((4'-chloro-[1,1'-biphenyl]-4-yl)methylene)-thiazolidine-2,4-dione-3-yl)acetamido)-3-(4-bromophenyl)propanamide (LJB-II-44) (A8); (S)-N-(3-Nitro-4-chlorobenzenesulfonyl)-2-(2-(5-((4'-chloro-[1,1'-biphenyl]-4-yl)methylene)-thiazolidine-2,4-dione-3-yl)acetamido)-3-(4-bromophenyl)propanamide (LJB-II-45) (A9); (S)-N-(4-Nitrobenzenesulfonyl)-2-(2-(5-((4'-methyl-[1,1'-biphenyl]-4-yl)methylene)-thiazolidine-2,4-dione-3-yl)acetamido)-3-(4-bromophenyl)propanamide (LJB-II-41) (A10); (S)-N-(3-Nitro-4-chlorobenzenesulfonyl)-2-(2-(5-(([1,1'-biphenyl]-4-yl)methylene)-thiazolidine-2,4-dione-3-yl)acetamido)-3-(4-bromophenyl)propanamide (DX-I-10) (A11).

2. The preparation method of the substituted 3-thiazolidine-2,4-dione Mcl-1 protein inhibitor according to claim 1, comprising the following steps: Step 1: Using thiazolidine-2,4-dione as the starting material, first performing a Knoevenagel condensation reaction with differently substituted formaldehydes to generate intermediates 1a-1c; Step 2: Performing a nucleophilic substitution reaction with methyl bromoacetate at the N position of the 3-position thiazolidine of the intermediate 1a-1c and removing the methyl ester under acidic conditions to generate key intermediates 2a-2c; Step 3: Performing an amide condensation reaction with differently substituted amino acids 3a-3e and differently substituted benzenesulfonamides to generate key intermediates 4a-4k; Step 4: The key intermediates 4a - 4k are first deprotected from Boc in ethyl acetate saturated with hydrogen chloride gas, and then, using the mixed anhydride method, undergo amide condensation reaction with another key intermediate 2a - 2c under the action of N - methylmorpholine and isobutyl chloroformate to obtain the target compound, which is a substituted 3 - thiazolidine - 2,4 - dione - type Mcl - 1 protein inhibitor.

3. The preparation method of the substituted 3-thiazolidine-2,4-dione Mcl-1 protein inhibitor according to claim 2, wherein: Reagents and conditions a: Different substituted formaldehydes, piperidine, ethanol, reflux.

4. The preparation method of the substituted 3-thiazolidine-2,4-dione Mcl-1 protein inhibitor according to claim 2, characterized in that: Reagents and conditions b: i. Methyl bromoacetate, potassium carbonate, acetone, reflux; ii. Glacial acetic acid, concentrated hydrochloric acid, reflux.

5. The preparation method of the substituted 3-thiazolidine-2,4-dione Mcl-1 protein inhibitor according to claim 2, wherein: Reagents and conditions c: 2 - (7 - azabenzotriazol - 1 - yl) - N,N,N',N' - tetramethyluronium hexafluorophosphate (HATU), N,N - diisopropylethylamine (DIEA), dichloromethane, ice bath - room temperature.

6. The preparation method of the substituted 3-thiazolidine-2,4-dione Mcl-1 protein inhibitor according to claim 2, characterized in that: Reagents and conditions d: i. Ethyl acetate solution saturated with hydrogen chloride, room temperature; ii. N - methylmorpholine, isobutyl chloroformate, tetrahydrofuran, - 20 °C - room temperature.

7. Use of the compound according to claim 1 in the preparation of a medicament for preventing or treating mammalian diseases caused by abnormal Mcl - 1 protein expression, wherein the mammalian diseases caused by abnormal Mcl - 1 protein expression are selected from myeloma and non - small cell lung cancer.

8. A pharmaceutical composition suitable for oral administration to mammals, comprising the compound according to claim 1 and one or more pharmaceutically acceptable carriers or excipients.

9. A pharmaceutical composition suitable for parenteral administration to mammals, comprising the compound according to claim 1 and one or more pharmaceutically acceptable carriers or excipients.

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