Aniline derivatives, processes for their preparation and pharmaceutical uses thereof

Abstract

The application relates to the technical field of cancer treatment drugs, and discloses an aniline derivative and a preparation method and pharmaceutical use thereof, as shown in a formula, wherein the substituents on a benzene ring of the compound are optimized, the compound has very strong proliferation inhibition activity on triple-negative breast cancer cells, and has great application potential.

Description

Technical Field

[0001] This invention relates to cancer treatment drugs, specifically to an aniline derivative, its preparation method, and its pharmaceutical uses. Background Technology

[0002] The statements in this section provide only background information relevant to the disclosure of this application and may not constitute prior art.

[0003] Breast cancer is the most common malignant tumor among women worldwide, seriously threatening their lives, health, and quality of life. In recent years, its incidence and mortality rates have been increasing year by year and showing a trend towards affecting younger women. Based on the molecular subtyping characteristics of breast cancer, it can be divided into subtypes such as luminal breast cancer, HER2-overexpressing breast cancer, and triple-negative breast cancer. Among them, triple-negative breast cancer (TNBC) is the most malignant and has the worst prognosis because it is negative for estrogen receptor (ER), progesterone receptor (PR), and human epidermal growth factor receptor 2 (HER2).

[0004] Compared to other breast cancer subtypes, triple-negative breast cancer is characterized by its high invasiveness, rapid proliferation, high rate of distant metastasis, high recurrence rate, and short survival. Furthermore, due to the lack of three major targeted therapeutic targets—ER, PR, and HER2—current mainstream clinical anti-tumor regimens such as endocrine therapy and HER2-targeted therapy are ineffective, significantly limiting the choice of clinical treatment methods. Currently, the core treatment regimens for triple-negative breast cancer still primarily consist of traditional chemotherapy, surgical resection, and radiotherapy. However, surgery is only suitable for early-stage localized lesions and cannot address tumor micrometastasis and the potential risk of recurrence. Radiotherapy and chemotherapy have drawbacks such as poor specificity, strong toxic side effects, and a high risk of tumor drug resistance. Many patients experience disease recurrence and distant metastasis after staged treatment, and long-term chemotherapy can lead to a series of complications such as immune damage and organ dysfunction, severely impacting patient prognosis.

[0005] Currently, there is a severe shortage of specific treatments for triple-negative breast cancer (TNBC) globally, and no highly effective, low-toxicity, and targeted therapy has yet achieved widespread clinical application. Existing candidate drugs generally suffer from weak antitumor activity, poor inhibition of TNBC tumor cell proliferation, single mechanisms of action, and high rates of drug resistance, failing to meet the core clinical needs for efficient treatment and effective prevention of TNBC. Therefore, developing novel active compounds that can efficiently inhibit the proliferation of TNBC tumor cells, possess high safety, and have both therapeutic and preventative potential is a key focus and hot topic in antitumor drug development in this field, with significant clinical value and broad application prospects.

[0006] MDA-MB-231 cells are a classic human triple-negative breast cancer cell line with stable TNBC malignant biological characteristics. They are currently the core model for screening and evaluating anti-tumor active compounds for triple-negative breast cancer in the laboratory, and can truly reflect the intervention effect of candidate drugs on triple-negative breast cancer.

[0007] Existing publicly available research shows that most anti-TNBC small molecule compounds suffer from drawbacks such as weak inhibitory activity, poor targeting, significant side effects, and a single mechanism of action. Most candidate compounds can only mildly inhibit tumor cell proliferation and cannot effectively block tumor cell invasion and metastasis. Furthermore, very few compounds offer both preventative and therapeutic applications. Therefore, developing a new class of small molecule compounds with the potential to both prevent and treat triple-negative breast cancer can overcome the shortcomings of existing technologies, providing novel drug candidates for prevention and intervention in high-risk groups and clinical treatment of diagnosed patients, thus possessing significant clinical application value and scientific research significance. Summary of the Invention

[0008] The purpose of this invention is to address the problems of low activity and limited variety of existing small molecule compounds by providing an aniline derivative, its preparation method, and its pharmaceutical uses. Through substituent optimization, a novel aniline derivative with excellent proliferative inhibitory activity against MDA-MB-231 triple-negative breast cancer cells was obtained. This derivative can effectively inhibit the proliferation of triple-negative breast cancer cells and suppress the malignant progression of tumors. It is expected to break through the technical bottleneck of existing therapeutic drugs and can be widely used in the prevention and clinical treatment of triple-negative breast cancer, providing a new candidate material basis for the development of targeted drugs for triple-negative breast cancer.

[0009] The technical solution of the present invention is as follows: In one aspect, the present invention provides an aniline derivative of Formula I or thereof, a stereoisomer, a tautomer, a pharmaceutically acceptable salt, a polymorph, a cocrystal, a solvate, a metabolite, a prodrug, or any mixture of two or more thereof:

[0010] In the formula: R1, R2, R3, and R4 are each independently selected from hydrogen, deuterium, halogen, hydroxyl group, CN, NO2, and C(O)R. a CO2R a C(O)NR c R d S(O)R b SO2R b SO2NR c R d NR c R d C1-C6 alkyl, C3-C7 cycloalkyl, C1-C6 alkoxy, C6-C14 aryl and 5-14 heteroaryl; optionally C1-C6 alkyl, C3-C7 cycloalkyl, C1-C6 alkoxy, C6-C 14 The aryl group and the 5-14 heteroaryl group are each separated by one or more R groups. e replace; R5 and R6 are each independently selected from hydrogen, C1-C6 alkyl, C3-C7 cycloalkyl, C6-C 14 Aryl, 3-8 membered heterocyclic and 5-14 membered heteroaryl, -C(O)R b SO2R b The C1-C6 alkyl, C3-C7 cycloalkyl, C6-C 14 Aryl, 3-8 membered heterocyclic and 5-14 membered heteroaryl are each optionally bounded by one or more R e replace; Alternatively, R5 and R6 together with the nitrogen atom they are attached to form a 4-7 membered heterocyclic group; R a Selected from hydrogen, C1-C6 alkyl, C3-C7 cycloalkyl, C6-C 14 Aryl and 5-14 heteroaryl groups; R b Selected from hydrogen, C1-C6 alkyl, C3-C7 cycloalkyl, C6-C 14 Aryl and 5-14 heteroaryl groups; R c and R d Each is independently selected from hydrogen, C1-C6 alkyl, C3-C7 cycloalkyl, and 4-7 membered heterocyclic groups; or R c and R d Together with the nitrogen atom it is attached to, it forms a 4-7 membered heterocyclic group; R e Selected from hydrogen, halogen, hydroxyl, CN, C(O)R g NR h R i C(O)NR h R i NR h C(O)R g C1-C6 alkyl, C1-C6 alkoxy, C3-C7 cycloalkyl or 3-8 membered heterocyclic groups; R g Selected from hydrogen, C1-C6 alkyl, C3-C7 cycloalkyl, C6-C 14 Aryl and 5-14 heteroaryl groups; R h and R i Each is independently selected from hydrogen, C1-C6 alkyl, C3-C7 cycloalkyl, C6-C 14 Aryl and 5-14 heteroaryl compounds.

[0011] According to a preferred embodiment, R1, R2, R3, and R4 are each independently selected from hydrogen, deuterium, halogen, hydroxyl, CN, NO2, and C(O)R. a CO2R a C(O)NR c R d NR c R d C1-C6 alkyl, C3-C7 cycloalkyl, C1-C6 alkoxy, C6-C 14 aryl and 5-14 heteroaryl groups, wherein the C1-C6 alkyl, C3-C7 cycloalkyl, C1-C6 alkoxy, C6-C 14 The aryl and 5-14 heteroaryl groups are each optionally bounded by one or more R groups. e replace; Preferably, R1, R2, R3, and R4 are each independently selected from hydrogen, deuterium, halogen, hydroxyl, CN, C1-C6 alkyl, C3-C7 cycloalkyl, C1-C6 alkoxy, C6-C 14 aryl and 5-14 heteroaryl groups, wherein the C1-C6 alkyl, C3-C7 cycloalkyl, C1-C6 alkoxy, C6-C 14 The aryl and 5-14 heteroaryl groups are each optionally bounded by one or more R groups. e replace; More preferably, R1, R2, R3, and R4 are each independently selected from hydrogen, deuterium, halogen, C1-C6 alkyl, C3-C7 cycloalkyl, and C1-C6 alkoxy; wherein the C1-C6 alkyl, C3-C7 cycloalkyl, and C1-C6 alkoxy are each optionally converted by one or more R e replace; More preferably, R1, R2, R3 and R4 are each independently selected from hydrogen, deuterium, halogen, C1-C6 alkyl, C3-C7 cycloalkyl, and C1-C6 alkoxy. More preferably, R1, R2, R3 and R4 are each independently selected from hydrogen and halogen; According to a preferred embodiment, R5 and R6 are each independently selected from hydrogen, C1-C6 alkyl, C3-C7 cycloalkyl, C6-C6 alkyl, C7 ... 14 aryl, 3-8 membered heterocyclic and 5-14 membered heteroaryl; the C1-C6 alkyl, C3-C7 cycloalkyl, C6-C 14 Aryl, 3-8 membered heterocyclic and 5-14 membered heteroaryl are each optionally bounded by one or more R e replace; Preferably, R5 and R6 are each independently selected from hydrogen, C1-C6 alkyl, C3-C7 cycloalkyl, C6-C6 alkyl, C7 ... 14 Aryl; the C1-C6 alkyl, C3-C7 cycloalkyl, C6-C14 Each aryl group is optionally bounded by one or more R e replace; More preferably, R5 and R6 are each independently selected from hydrogen, C1-C6 alkyl, C3-C7 cycloalkyl, C6-C6 alkyl, C7 ... 14 Aryl; the C1-C6 alkyl, C3-C7 cycloalkyl, C6-C 14 Each aryl group is optionally bounded by one or more R e replace; More preferably, R5 and R6 are each independently selected from hydrogen and C1-C6 alkyl groups; the C1-C6 alkyl group is affected by one or more R... e replace; Alternatively, R5 and R6 together with the nitrogen atom they are attached to form a 4-7 membered heterocyclic group; According to a preferred embodiment, R a Selected from hydrogen, C1-C6 alkyl, C3-C7 cycloalkyl, C6-C 14 Aryl and 5-14 heteroaryl groups; Preferably, R a Independently selected from hydrogen and C1-C6 alkyl groups; According to a preferred embodiment, R b Selected from C1-C6 alkyl, C3-C7 cycloalkyl, C6-C 14 Aryl and 5-14 heteroaryl groups; Preferably, R b Selected from hydrogen and C1-C6 alkyl groups; According to a preferred embodiment, R c and R d Each is independently selected from hydrogen, C1-C6 alkyl, C3-C7 cycloalkyl, and 4-7 membered heterocyclic groups; Preferably, R c and R d Each is independently selected from hydrogen and C1-C6 alkyl groups; Or R c and R d Together with the nitrogen atom it is attached to, it forms a 4-7 membered heterocyclic group; According to a preferred embodiment, R e Independently selected from hydrogen, C(O)R g NR h R i C(O)NR h R i NR h C(O)R g C1-C6 alkyl, C1-C6 alkoxy, C3-C7 cycloalkyl, 3-8 membered heterocyclic groups; Preferably, R eIndependently selected from hydrogen, NR h R i C1-C6 alkyl, C1-C6 alkoxy, C3-C7 cycloalkyl, 3-8 membered heterocyclic groups.

[0012] More preferably, R e Independently selected from hydrogen, NR h R i C1-C6 alkyl, C1-C6 alkoxy.

[0013] According to a preferred embodiment, R g Independently selected from hydrogen, C1-C6 alkyl, C3-C7 cycloalkyl, C6-C 14 Aryl and 5-14 heteroaryl groups; Preferably, R g It is independently selected from hydrogen and C1-C6 alkyl groups.

[0014] According to a preferred embodiment, R h and R i Each is independently selected from hydrogen, C1-C6 alkyl, C3-C7 cycloalkyl, C6-C 14 Aryl and 5-14 heteroaryl groups; Preferably, R h and R i Each is independently selected from hydrogen and C1-C6 alkyl groups.

[0015] According to a preferred embodiment, R1, R2, R3 and R4 are each independently selected from hydrogen and halogens; R5 and R6 are each independently selected from hydrogen and C1-C6 alkyl groups; the C1-C6 alkyl groups are separated by one or more R5 and R6. e replace; Alternatively, R5 and R6 together with the nitrogen atom they are attached to form a 4-7 membered heterocyclic group; R e Independently selected from hydrogen, NR h R i C1-C6 alkyl, C1-C6 alkoxy; R h and R i Each is independently selected from hydrogen and C1-C6 alkyl groups.

[0016] According to a preferred embodiment, an aniline derivative is selected from the following compounds: ; Compounds 1-8 are respectively: (1)(S)-2-[(R)-3-(3,5-dichloro-4-(piperidin-1-yl)phenyl)-2-[(2S,3S)-3-methyl-2-(methylamino)pentamido]propamido]-3-methyl-3-((triethylsilyl)oxy)methyl butyrate; (2)(S)-2-[(R)-3-(3-chloro-4-(piperidin-1-yl)phenyl)-2-[(2S,3S)-3-methyl-2-(methylamino)pentamido]propamido]-3-methyl-3-((triethylsilyl)oxy)methyl butyrate; (3)(S)-2-[(R)-3-(4-(dimethylamino)phenyl)-2-[(2S,3S)-3-methyl-2-(methylamino)pentamido]propamido]-3-methyl-3-((triethylsilyl)oxy)methyl butyrate; (4)(S)-2-[(R)-3-(4-dimethylamino-2-fluorophenyl)-2-[(2S,3S)-3-methyl-2-(methylamino)pentamido]propamido]-3-methyl-3-(triethylsiloxy)butyrate; (5)(S)-2-[(R)-3-(3,5-dichloro-4-dimethylaminophenyl)-2-[(2S,3S)-3-methyl-2-(methylamino)pentamido]propamido]-3-methyl-3-(triethylsiloxy)butyrate; (6) (S)-2-[(R)-3-(3,5-dichloro-4-(pyrrolidin-1-yl)phenyl)-2-[(2S,3S)-3-methyl-2-(methylamino)pentamido]propamido]-3-methyl-3-(triethylsiloxy)butyrate; (7)(S)-2-[(R)-3-(3,5-dichloro-4-morpholinylphenyl)-2-[(2S,3S)-3-methyl-2-(methylamino)pentamido]propamido]-3-methyl-3-(triethylsiloxy)butyrate; (8)(S)-2-[(R)-3-(3,5-dichloro-4-(4-methylpiperazin-1-yl)phenyl)-2-[(2S,3S)-3-methyl-2-(methylamino)pentamido]propamido]-3-methyl-3-(triethylsiloxy)butyrate.

[0017] According to a preferred embodiment, the steps include: Step 1: SM-1 is converted into IM-1 through N-alkylation and other reactions. Step 2: IM-1 reacts with zinc reagent to generate IM-2 under the action of Pd catalyst. Step 3: Hydrolysis of IM-2 to generate IM-3 Step 4: IM-3 and SM-3 react with a condensing agent to form IM-4. Step 5: IM-4 loses its Boc protecting group to generate IM-5. Step 6: IM-5 and SM-4 react with a condensing agent to form IM-6. Step 7: IM-6 undergoes TESOTf and 2,6-lutidine treatment to add a TES group to the hydroxyl group and simultaneously remove the Boc protecting group to obtain the aniline derivative of formula I; R1, R2, R3, R4, R5, and R6 are as described above; .

[0018] Another aspect of the present invention provides a pharmaceutical composition for treating cancer, comprising the tripeptide compound as described above.

[0019] Preferably, it also includes one or more pharmaceutically acceptable carriers.

[0020] Another aspect of the present invention provides a pharmaceutical formulation for treating cancer, comprising the tripeptide compound as described above. The prodrug comprises an ester compound; the stereoisomer comprises a racemic compound.

[0021] Preferably, the pharmaceutical formulation further includes pharmaceutically acceptable excipients for preparing solid, semi-solid, liquid, or gaseous formulations.

[0022] Another aspect of the present invention provides the use of the aniline derivatives as described above and the pharmaceutical compositions for treating cancer as described above in the preparation of medicaments for improving, preventing, and treating triple-negative breast cancer.

[0023] Compared with existing technologies, the advantages of this invention are: 1. An aniline derivative and its pharmaceutical use. Cell experiments have confirmed that the aniline derivatives 1, 3, 4, 5, and 6 of this application have an inhibitory effect on the proliferation of MDA-MB-231 cells at a drug concentration of 2 μM, especially compounds 3 and 4; the cell survival rate is only 20% of that of the control group, showing a significant inhibitory effect; confirming that the compounds of this application can inhibit the development of triple-negative breast cancer.

[0024] 2. An aniline derivative and its preparation method, wherein the inhibitory activity of the original compound against triple-negative breast cancer is enhanced by modifying and optimizing the substituents of the original structural formula; the IC50 values ​​of compounds 3 and 4 are... 50 The value is below 2 μM; using the preparation method and substituent settings of this application, novel small molecule drugs with the potential to improve, prevent or treat triple-negative breast cancer can be prepared. Detailed Implementation

[0025] The specific embodiments listed in this invention are merely examples, and the invention is not limited to the specific embodiments described below. For those skilled in the art, any equivalent modifications and substitutions to the embodiments described below are also within the scope of this invention. Therefore, all equivalent transformations and modifications made without departing from the spirit and scope of this invention should be covered within its scope. Where specific conditions are not specified in the embodiments, conventional conditions or conditions recommended by the manufacturer shall apply. All reagents or instruments whose manufacturers are not specified are commercially available conventional products. To better illustrate this invention, numerous specific details are provided in the following detailed embodiments. Those skilled in the art should understand that this invention can be practiced even without certain specific details. In other embodiments, methods, means, equipment, and steps well known to those skilled in the art are not described in detail in order to highlight the main points of this invention.

[0026] Unless otherwise defined, all technical and scientific terms used herein have the same meanings as commonly understood by one of ordinary skill in the art. Unless otherwise specified, all units used in this specification are International Standard Units (SI), and all numerical values ​​and ranges appearing in this invention should be understood to include systematic errors unavoidable in industrial production.

[0027] General terms and definitions Unless otherwise defined, the terms used herein have the same meaning as commonly understood by those skilled in the art. The technical intent used herein refers to technology commonly understood in the art, including variations or equivalent substitutions that are obvious to those skilled in the art. While the following terms are readily understood by those skilled in the art, they are set forth below to better explain the invention.

[0028] The term "isomer" refers to a compound that has the same molecular weight due to having the same number and type of atoms, but with different spatial arrangements or configurations of atoms.

[0029] The term "stereoisomer" (or "optical isomer") refers to a stable isomer that, due to having at least one chiral element (including a chiral center, chiral axis, chiral plane, etc.), has a perpendicular asymmetric plane, thereby enabling the rotation of plane-polarized light. Since the compounds of the present invention (or pharmaceutically acceptable salts thereof) comprise asymmetric carbon atoms, they can exist as a single stereoisomer, a racemic mixture, or a mixture of enantiomers and diastereomers. These compounds can be monomorphic compounds, mixtures of racemic mixtures, or mixtures of a single stereoisomer enriched with other enantiomers and / or diastereomers.

[0030] The term "tautomer" (or "tautomer form") refers to structural isomers with different energies that can interconvert through a low energy barrier. If tautomerism is possible (e.g., in solution), chemical equilibrium can be achieved in the tautomer. For example, proton tautomers (or proton transfer tautomers) include (but are not limited to) interconversions via proton transfer, such as keto-enol isomerization, imine-enamine isomerization, amide-imine alcohol isomerization, etc. Unless otherwise stated, all tautomer forms of the compounds of this invention are within the scope of this invention.

[0031] The term "pharmaceutically acceptable salt" refers to a salt of the compound of the present invention that is substantially non-toxic to organisms. Pharmaceutically acceptable salts generally include (but are not limited to) salts formed by the reaction of the compound of the present invention with a pharmaceutically acceptable inorganic acid / organic acid / acidic amino acid or inorganic base / organic base / basic amino acid; such salts are also known as acid addition salts or base addition salts.

[0032] The term "polymorph" (or "polymorphic form") refers to the solid crystalline form of a compound or complex. Those skilled in the art can obtain polymorphs of molecules using many known methods. These methods include (but are not limited to) melt recrystallization, melt cooling, solvent recrystallization, desolvation, rapid evaporation, rapid cooling, slow cooling, vapor diffusion, and sublimation. Furthermore, well-known techniques can be used to detect, classify, and identify polymorphs, including (but not limited to) differential scanning calorimetry (DSC), thermogravimetric analysis (TGA), X-ray powder diffraction (XRPD), single-crystal X-ray diffraction (SCXRD), solid-state nuclear magnetic resonance (NMR), infrared spectroscopy (IR), Raman spectroscopy, and scanning electron microscopy (SEM). This invention covers all possible crystalline forms or polymorphs of the compounds of this invention, which can be a single polymorph or a mixture of multiple polymorphs in any proportion.

[0033] The term "eutectic" refers to a multi-component molecular crystal complex formed in the solid state by spontaneous association of two or more chemically independent pure components through non-covalent intermolecular interactions (including hydrogen bonds, van der Waals forces, π-π stacking interactions, dipole interactions, etc.) without the breaking or formation of ionic or covalent bonds. Each component of the complex retains its original chemical molecular structure, without proton transfer or salt bond formation, and the components are regularly arranged in the same lattice system with a fixed stoichiometric ratio. This is distinct from physical mixtures, solvates, and polymorphs, and possesses inherent characteristics such as melting point, solubility, dissolution characteristics, physicochemical stability, and powder properties that differ from those of the individual components.

[0034] The term "solvent" refers to a substance formed by the combination of a compound of the present invention (or a pharmaceutically acceptable salt thereof) with at least one solvent molecule through non-covalent intermolecular forces. Common solvates include (but are not limited to) hydrates (including hemihydrates, monohydrates, dihydrates, trihydrates, etc.), ethanol compounds, acetone compounds, etc. The compounds of the present invention may exist in the form of solvates (preferably hydrates), containing a polar solvent (particularly water, methanol, or ethanol) as a lattice structure element. The amount of polar solvent (particularly water) may be present in stoichiometric or non-stoichiometric form.

[0035] The term "metabolite" refers to a derivative compound formed after the compounds of the present invention are metabolized, for example, by reactions such as oxidation, reduction, hydrolysis, amidation, deamidation, esterification, or enzymatic hydrolysis. The present invention covers all possible metabolite forms of the compounds of the present invention, i.e., substances formed in the body of an individual administering the compounds of the present invention. Metabolites of the compounds can be identified using techniques known in the art, and their activity can be characterized experimentally.

[0036] The term "prodrug" refers to a derived compound that, upon administration to an individual, can directly or indirectly provide the compounds of the present invention. Particularly preferred derived compounds or prodrugs are those that, upon administration to an individual, can improve the bioavailability of the compounds of the present invention (e.g., facilitate absorption into the bloodstream) or promote the delivery of the parent compound to its site of action (e.g., the lymphatic system). Unless otherwise indicated, all prodrug forms of the compounds of the present invention are within the scope of the present invention. Furthermore, the present invention also covers compounds of the present invention containing protecting groups. In any process of preparing the compounds of the present invention, protection of sensitive or reactive groups on any relevant molecule may be necessary and / or desired, thereby forming a form of chemical protection for the compounds of the present invention. This can be achieved by conventional protecting groups. These protecting groups can be removed at appropriate subsequent stages using methods known in the art.

[0037] The terms “including,” “comprising,” “having,” “containing,” or “involving,” and their other variations herein, are inclusive or open-ended and do not exclude other unlisted elements or method steps.

[0038] The term "halogenated" or "halogenated" is defined as including F, Cl, Br, or I.

[0039] The term "alkyl" is defined as a straight-chain or branched saturated aliphatic hydrocarbon group. In some embodiments, the alkyl group has 1 to 12 carbon atoms, preferably 1 to 6 carbon atoms, and more preferably 1 to 4 carbon atoms. For example, as used herein, the term "C1-C6 alkyl" refers to a linear or branched group with 1 to 6 carbon atoms (e.g., methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, or n-hexyl). In this invention, the "alkyl" group is optionally substituted with one or more (such as 1 to 3) suitable substituents such as halogens (in which case the group is referred to as "haloalkyl") (e.g., -CF3, -C2F5, -CHF2, -CH2F, -CH2CF3, -CH2Cl, or -CH2CH2CF3, etc.).

[0040] The term “cycloalkyl” refers to a monocyclic or polycyclic (e.g., bicyclic) saturated or partially unsaturated aliphatic monovalent hydrocarbon group (e.g., monocyclic cycloalkyl, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl, etc.; or bicyclic cycloalkyl, including fused ring, bridged ring or spiro ring, such as decahydronaphthyl, bicyclic [2.2.1]heptyl, spiro [4.5]decyl, etc.).

[0041] The term "heterocyclic alkyl" refers to a monocyclic or polycyclic (e.g., bicyclic) saturated or partially unsaturated aliphatic monovalent hydrocarbon group having one or more carbon atoms (e.g., 1, 2, 3, 4, 5, 6, 7, 8, or 9) and one or more (e.g., 1, 2, 3, or 4) segments each independently selected from O, S, S(=O), S(=O)₂, and NR (R represents a hydrogen atom or substituent, such as alkyl or cycloalkyl). The ring system in a heterocyclic alkyl group can be a fused ring, a bridged ring, or a spirocyclic system. If the valence requirements are met, a heterocyclic alkyl group can be linked to other groups (or segments) through any carbon atom or heteroatom in the ring.

[0042] The term "alkenyl" refers to a hydrocarbon group containing at least one carbon-carbon double bond. Alkenyl groups can be straight-chain or branched.

[0043] The term "alkynyl" refers to a hydrocarbon group containing at least one carbon-carbon triple bond. Alynyl groups can be straight-chain or branched.

[0044] The term "aryl" refers to a monocyclic or fused-ring aromatic group with a conjugated π-electron system that is entirely carbon. For example, as used herein, the term "C6-C" is used in the following context.14 "Aryl" refers to an aromatic group containing 6 to 14 carbon atoms, such as phenyl or naphthyl. The aryl group may optionally be replaced by one or more (such as 1 to 3) suitable substituents (e.g., halogen, -OH, -CN, C1-C6 alkyl, etc.).

[0045] The term "heteroaryl" refers to a monocyclic or polycyclic aromatic ring system containing at least one heteroatom, which may be the same or different (the heteroatom is, for example, oxygen, nitrogen, or sulfur), and, in some cases, may be benzo-fused. In some embodiments, the heteroaryl group has 3 to 18 cyclic atoms, preferably 5 to 10. In particular, the heteroaryl group is selected from thiophene, furanyl, pyrrole, oxazolyl, thiazolyl, imidazolyl, pyrazolyl, isoxazolyl, isothiazolyl, oxadiazolyl, triazolyl, thiazolyl, etc., and their benzo-derived derivatives; or pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, triazinyl, etc., and their benzo-derived derivatives.

[0046] The term "substitution" refers to the replacement of one or more (e.g., 1, 2, 3, or 4) hydrogen atoms on a specified atom by a selected group, provided that the substitution does not exceed the normal valence of the specified atom in its current state and the substitution forms a stable compound. The number of substituted groups selected is permitted only when such a combination forms a stable compound.

[0047] If a substituent is described as “each independently selected”, then each substituent may be the same as or different from another (other) substituent. The term “one or more” means one or more under reasonable conditions, such as two, three, four, five, or ten.

[0048] This invention also covers compounds of the invention containing protecting groups. In any process of preparing the compounds of the invention, protection of sensitive or reactive groups on any relevant molecule may be necessary and / or desired, thereby forming a form of chemical protection for the compounds of the invention.

[0049] The term "protecting group" refers to a chemical group that has the following characteristics: 1) Can react with corresponding functional groups to form protected groups. 2) The protected groups will be stable under the reaction conditions. 3) Functional groups can be released by desorption from protected groups. In addition, groups not defined herein follow their usual definitions. Pharmaceutically acceptable salts of the compounds of this invention include their acid addition salts and base addition salts. Examples include salts formed from alkali metals, alkaline earth metals, ammonium, alkylammonium, etc., and salts formed with inorganic or organic acids. These salts can be listed as sodium salts, potassium salts, calcium salts, ammonium salts, aluminum salts, triethylammonium salts, acetates, propionates, butyrates, formates, trifluoroacetates, maleates, tartrates, citrates, stearates, succinates, ethylsuccinates, lactobionates, gluconates, glucohepanoates, benzoates, methanesulfonates, ethanesulfonates, 2-hydroxyethanesulfonates, benzenesulfonates, p-toluenesulfonates, lauryl sulfates, malates, aspartate salts, glutamates, adipates, tris(hydroxymethyl)aminomethane salts, salts formed with cysteine, salts formed with N-acetylcysteine, hydrochlorides, hydrobromide salts, phosphates, sulfates, hydroiodates, nicotinate salts, oxalates, picrates, thiocyanates, undecanoates, salts formed with acrylic acid polymers, and salts formed with carboxyl vinyl polymers, etc.

[0050] The compounds of the present invention can exist as solvates (preferably hydrates), wherein the compounds of the present invention contain a polar solvent, particularly, for example, water, methanol, or ethanol, as a structural element of the lattice of the compound. The amount of the polar solvent, particularly water, can be stoichiometric or non-stoichiometric.

[0051] The scope of this invention also includes metabolites of the compounds of this invention, i.e., substances formed in the body when the compounds of this invention are administered. Such products can be generated, for example, by oxidation, reduction, hydrolysis, amidation, deamidation, esterification, defatting, enzymatic hydrolysis, etc., of the administered compound. Therefore, this invention includes metabolites of the compounds of this invention, including compounds obtained by methods that expose the compounds of this invention to mammals for a time sufficient to produce their metabolites.

[0052] The present invention further includes, within its scope, prodrugs of the compounds of the present invention, which are certain derivatives of the compounds of the present invention that may themselves have little or no pharmacological activity, and which, when administered to or onto the body, can be converted, for example, by hydrolysis and cleavage into the compounds of the present invention having the desired activity. Typically, such prodrugs are functional group derivatives of the compounds that are readily converted in vivo into the desired therapeutically active compounds.

[0053] The features and performance of the present invention will be further described in detail below with reference to embodiments.

[0054] Example 1: Preparation of an aniline derivative The preparation of compound 1 includes the following steps: Step 1: 2,6-Dichloro-4-iodoaniline (1-2) Compound 1-1 (9.72 g, 60.0 mmol) was dissolved in 100 mL of acetonitrile, and then NIS (16.2 g, 72 mmol) and 100 mL of acetic acid were added. The mixture was heated to 70 °C and reacted for 2 hours. The reaction solution was concentrated to remove most of the acetonitrile and acetic acid. Then 200 mL of water was added, and the pH was adjusted to about 8 with saturated sodium bicarbonate solution. A large amount of off-white solid precipitated out. The mixture was stirred thoroughly, filtered, and the filter cake was washed with a small amount of water and dried to obtain compound 1-2 (17.5 g, yield: 98.3%).

[0055] MS m / z (ESI): 287.9 ​​[M+H] + .

[0056] Step 2: 1-(2,6-dichloro-4-iodophenyl)piperidine (1-3) Compounds 1-2 (2.88 g, 10.0 mmol) were dissolved in 30 mL of DMSO and cooled to 10 °C. 60% NaH (1.6 g, 40.0 mmol) was added, and the mixture was stirred for 20 minutes. Then, 1,5-dibromopentane (2.76 g, 12.0 mmol) was added, and the mixture was stirred at 10 °C for 4 hours. The reaction mixture was quenched with 30 mL of water and extracted three times with 50 mL of EA solution. The organic phases were combined and washed twice with 50 mL of saturated sodium chloride solution. The organic phase was concentrated, and 30 mL of water was added to the residue. The mixture was stirred and slurried in an ice bath for 2 hours. The mixture was filtered, the filter cake was washed with water, and dried to give a yellow solid product 1-3 (2.6 g, yield: 73%).

[0057] MS m / z (ESI): 355.9 [M+H] + .

[0058] Step 3: (R)-2-[(tert-Butoxycarbonyl)amino]-3-[3,5-Dichloro-4-(piperidin-1-yl)phenyl]methyl propionate (1-4) Under nitrogen protection, 1-3 (2.16 g, 6.1 mmol) was dissolved in 25 mL of DMF, followed by Pd2(dba)3 (280 mg, 0.3 mmol) and tris(2-methylphenyl)phosphine (182 mg, 0.6 mmol), and finally 1-4a (2.87 g, 7.3 mmol) in 20 mL of DMF. The mixture was heated to 60 °C and stirred overnight. The reaction solution was cooled to room temperature, 50 mL of water was added, and the mixture was stirred thoroughly. The solution was extracted twice with 50 mL of EA, and the organic phases were combined and washed twice with saturated sodium chloride. The organic phase was concentrated and purified using a reversed-phase preparative column to give a white solid product 1-4 (680 mg, yield: 26.1%).

[0059] MS m / z (ESI): 430.1 [M+H] + .

[0060] Step 4: (R)-2-[(tert-Butoxycarbonyl)amino]-3-[3,5-Dichloro-4-(piperidin-1-yl)phenyl]propionic acid (1-5) Compounds 1-4 (680 mg, 1.57 mmol) were dissolved in a mixed solvent of 7.5 mL THF and 2.5 mL water, and then LiOH·H2O (130 mg, 3.15 mmol) was added. The mixture was stirred overnight at room temperature. The pH of the reaction solution was adjusted to about 5 with 1 N HCl, and 20 mL water and 20 mL EA were added. After thorough stirring, the mixture was separated. The aqueous phase was extracted once with EA. The organic phases were combined and concentrated. The residue was purified by reversed-phase preparative column chromatography to obtain a white solid product 1-5 (516 mg, 78.3%).

[0061] MS m / z (ESI): 416.1 [M+H] + .

[0062] Step 5: (S)-2-[(R)-2-(tert-butoxycarbonylamino)-3-(3,5-dichloro-4-(piperidin-1-yl)phenyl)propionamido]-3-hydroxy-3-methylbutyrate (1-6) Compounds 1-5 (80 mg, 0.19 mmol) were dissolved in 5 mL of DCM, and HATU (88 mg, 0.23 mmol) and DIPEA (50 mg, 0.38 mmol) were added, followed by 1-6a (30 mg, 0.23 mmol). The mixture was stirred at room temperature for 2 hours. The reaction solution was quenched with 20 mL of water, extracted twice with 20 mL of DCM, and the organic phases were combined and concentrated. The residue was purified by reversed-phase preparative column chromatography to give a white solid product 1-6 (92 mg, 84.2%).

[0063] MS m / z (ESI): 546.2 [M+H] + .

[0064] Step 6: (S)-2-[(R)-2-amino-3-(3,5-dichloro-4-(piperidin-1-yl)phenyl)propamido]-3-hydroxy-3-methylbutyrate (1-7) Compounds 1-6 (46 mg, 0.084 mmol) were dissolved in 10 mL of DCM, and then 5 mL of TFA was added. The mixture was stirred at room temperature for 2 hours. The reaction solution was then concentrated and used directly in the next step.

[0065] MS m / z (ESI): 446.1 [M+H] + .

[0066] Step 7: (6S,9R,12S)-6-((S)-sec-butyl)-9-(3,5-dichloro-4-(piperidin-1-yl)benzyl)-12-(2-hydroxypropyl-2-yl)-2,2,5-trimethyl-4,7,10-trioxo-3-oxa-5,8,11-triazatridecane-13-methyl ester (1-8) Dissolve 1-8a (20 mg, 0.084 mmol) in 5 mL of DCM, then add HATU (38 mg, 0.1 mmol) and DIPEA (22 mg, 0.168 mmol). Subsequently, dissolve the crude product from the previous step in approximately 5 mL of DCM and add it to the reaction solution. Stir at room temperature for 2 hours. Quench the reaction solution with saturated ammonium chloride solution. Extract twice with DCM, combine the organic phases and concentrate. Purify the residue using a reverse-phase preparative column to obtain a white solid product 1-8 (45 mg, combined yield of two steps: 79.7%).

[0067] MS m / z (ESI): 673.3 [M+H] + .

[0068] Step 8: (S)-2-[(R)-3-(3,5-dichloro-4-(piperidin-1-yl)phenyl)-2-[(2S,3S)-3-methyl-2-(methylamino)pentamido]propamido]-3-methyl-3-((triethylsilyl)oxy)methyl butyrate (1) Dissolve 1-8 (45 mg, 0.067 mmol) in DCM (5 mL), cool to 0–5 °C under nitrogen protection, then add 2,6-lutidine (0.5 mL) and TESOTf (0.5 mL) sequentially, and stir at 0–5 °C for 6 hours. Quench the reaction solution with 10 mL of saturated ammonium chloride solution, stir thoroughly, and separate the layers. Extract the aqueous phase twice with 10 mL of DCM solution, combine the organic phases and concentrate. Purify the residue using a reversed-phase preparative column to give a white solid product 1 (23 mg, yield: 50.1%).

[0069]

[0070] 1 H NMR (400 MHz, CDCl3) δ 7.63 (d, J = 8.0 Hz, 1H), 7.13 (s, 2H), 6.87 (d, J = 8.8 Hz, 1H), 4.69 – 4.54 (m, 1H), 4.36 (d, J = 8.8 Hz, 1H), 3.66(s, 3H), 3.17 – 2.99 (m, 5H), 2.89 (dd, J = 13.9, 7.6 Hz, 1H), 2.80 (d, J =4.5 Hz, 1H), 2.19 (s, 3H), 1.79 – 1.71 (m, 1H), 1.70 – 1.61 (m, 4H), 1.59 –1.54 (m, 2H), 1.46 – 1.39 (m, 1H), 1.32 (s, 3H), 1.17 – 1.05 (m, 4H), 0.94 –0.89 (m, 15H), 0.57 – 0.51 (m, 6H). MS m / z (ESI): 687.3 [M+H] + .

[0071] Example 2 Example 2 is a further improvement on Example 1; the difference between Example 2 and Example 1 is that, depending on the different substituents on the benzene ring of the product, SM-2 with different substituents is added in the third step; the remaining steps are the same, and compound 2-8 is prepared. The name of compound 2-8 and its NMR and mass spectrometry characterization data are shown in Table 1 below.

[0072] Table 1. Names and characterization data of some compounds

[0073] Test Example 1: In vitro screening experiment for single-concentration compound against breast cancer cell proliferation activity This experiment used the CCK-8 cell proliferation assay to test the in vitro inhibitory effects of a series of small molecule compounds on the proliferation of human breast cancer MDA-MB-231 cells under fixed drug concentration conditions, and to detect cell viability after compound treatment for preliminary activity screening of the compounds.

[0074] 1. Experimental Reagents and Instruments 1.1 Experimental cells: Human breast cancer cell line MDA-MB-231, purchased from Wuhan Pronosei Life Science Technology Co., Ltd.; 1.2 Detection kit: CCK-8 cell proliferation assay kit (Selleck, catalog number: B34302); 1.3 Experimental instruments: SpectraMax M5e multi-functional microplate reader (Molecular Devices).

[0075] 2. Cell Culture Conditions MDA-MB-231 cells are adherent cells, cultured in L-15 complete medium containing 10% fetal bovine serum (FBS) under CO2-free incubator conditions. After routine subculturing, cells in the logarithmic growth phase and in good growth condition were selected for experiments.

[0076] 3. Experimental Procedures 3.1 MDA-MB-231 cells in the logarithmic growth phase were seeded into 96-well cell culture plates and cultured overnight in an incubator. When the cell confluence reached 70% to 80%, the cells were treated with the drug.

[0077] 3.2 Prepare working solutions of the test compounds, with a uniform final concentration of 2 μM, using complete culture medium for dilution; set up blank control group (containing only culture medium, no cells) and negative control group (containing cells, no compound added), and set up 3 replicates for all test compound groups, using complete culture medium throughout the process.

[0078] 3.3 Discard the original culture medium in the wells, add the corresponding working solution of the test compound, and incubate continuously in a CO2-free incubator at 37 ℃ for 48 h.

[0079] 3.4 After incubation: Mix the CCK-8 stock solution with serum-free L-15 medium at a volume ratio of 1:9.

[0080] 3.5 Discard the drug solution in each well, add 100 μL of freshly prepared CCK-8 working solution to each well, and incubate at a constant temperature of 37 ℃ in the dark for 1~4 h.

[0081] 3.6 When the negative control solution turns a uniform orange-yellow color, remove the culture plate and use an ELISA reader to detect the absorbance (OD) value of each well at a wavelength of 450 nm.

[0082] 4. Data Processing and Result Determination 4.1 Formula for Calculating Cell Viability Formula for calculating cell viability: Cell viability (%) = [A (Drug+) – A (Black)] / [A (Drug-) – A (Black)]x 100% In the formula: A (drug administration well): OD value of the experimental well containing cells, 2 μM of the test compound, and CCK-8 working solution; A (Control Well): Negative control well containing cells, no compounds, and only CCK-8 working solution added; OD value. A (Blank Well): OD value of blank wells containing only culture medium and CCK-8 working solution and no cells.

[0083] 4.2 Data Analysis The cell viability of each test compound at a concentration of 2 μM was calculated using Excel. The cell viability was used to evaluate the inhibitory activity of the compounds on the proliferation of MDA-MB-231 cells. The lower the viability, the stronger the in vitro antitumor proliferative activity of the compounds.

[0084] 5. Test Results Table 2. Results of the inhibitory effect of the compound on the proliferation of MDA-MB-231 cells at a drug concentration of 2 μM.

[0085] As shown in Table 2, compounds 1, 3, 4, 5, and 6 of this application all exhibit good inhibitory effects on the proliferation of triple-negative breast cancer cells. In particular, compounds 3 and 4 of this application achieved an inhibition rate of approximately 80% against triple-negative breast cancer cells at a concentration of 2 μM, with an IC50 value of [missing information].50 The value is significantly lower than 2 μM, exhibiting extremely high MDA-MB-231 cell proliferation inhibitory activity, which can be used to prepare small molecule drugs for triple-negative breast cancer.

[0086] The embodiments described above merely illustrate specific implementation methods of this application, and while the descriptions are detailed and specific, they should not be construed as limiting the scope of protection of this application. It should be noted that those skilled in the art can make various modifications and improvements without departing from the concept of the technical solution of this application, and these modifications and improvements all fall within the scope of protection of this application.

Claims

1. An aniline derivative of formula I or a pharmaceutically acceptable salt: Specifically selected from the following compounds: 。 2. The method for preparing an aniline derivative according to claim 1, characterized in that, Includes the following steps: Step 1: SM-1 is converted into IM-1 through N-alkylation and other reactions. Step 2: IM-1 reacts with zinc reagent to generate IM-2 under the action of Pd catalyst. Step 3: Hydrolysis of IM-2 to generate IM-3 Step 4: IM-3 and SM-3 react with a condensing agent to form IM-4. Step 5: IM-4 loses its Boc protecting group to generate IM-5. Step 6: IM-5 and SM-4 react with a condensing agent to form IM-6. Step 7: IM-6 undergoes TESOTf and 2,6-lutidine treatment to add a TES group to the hydroxyl group and simultaneously remove the Boc protecting group to obtain the aniline derivative of formula I; Wherein, R1, R2, R3, R4, R5 and R6 are as described in claim 1; 。 3. A pharmaceutical composition for treating triple-negative breast cancer, characterized in that, Includes aniline derivatives as described in claim 1 or pharmaceutically acceptable salts.

4. The use of the aniline derivative or pharmaceutically acceptable salt of claim 1, or the pharmaceutical composition of claim 3 for treating triple-negative breast cancer, in the preparation of a medicament for treating triple-negative breast cancer.

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