Ferucic acid hydrazide compounds and their uses

Abstract

This invention belongs to the field of medicinal chemistry and relates to ferulate hydrazide compounds and their uses. Specifically, this invention provides compounds of formula I or pharmaceutically acceptable salts, stereoisomers, tautomers, polymorphs, solvates, racemates, prodrugs, or metabolites thereof, which exhibit good antitumor activity.

Description

Technical Field

[0001] This invention belongs to the field of medicinal chemistry and relates to a ferulic acid hydrazide compound with antitumor activity, a pharmaceutical composition containing the same, and its uses. Background Technology

[0002] Cancer is a malignant disease with extremely high incidence and mortality rates worldwide, and relatively poor treatment outcomes. According to research by the World Health Organization, there were 19.29 million new cancer cases globally in 2020, with 45.73 million in China, accounting for 23.7% of the global total, ranking first and approximately twice that of the United States, which ranked second in terms of new cancer cases. In 2020, breast cancer became the most common cancer globally, followed by lung cancer, but lung cancer is the leading cause of cancer death worldwide. Colorectal cancer is the second most common, ranking third in incidence, and its mortality rate is second only to lung cancer. Globally, anti-tumor drugs have become the largest therapeutic area in the pharmaceutical market. Currently, commonly used anti-tumor drugs include cytotoxic drugs, hormones, molecularly targeted therapies, biological response modifiers, tumor differentiation inducers, tumor angiogenesis inhibitors, and adjuvant therapies for tumors. Therefore, the discovery and development of anti-tumor drugs has always been a key focus of research for scientists worldwide and a major health and survival problem urgently needing to be solved.

[0003] Abnormalities in intracellular apoptosis signaling pathways in tumor cells often allow them to escape apoptosis induced by endogenous factors or exogenous drugs, leading to tumor development or drug resistance. Procaspase-3 is an important downstream executive protein in the intracellular apoptosis signaling pathway and is expressed at elevated levels in various malignant tumors. In 2006, foreign researchers first reported that PAC-1 (procaspase activating compound 1) can exert anti-tumor effects by activating procaspase-3, exhibiting growth inhibition against tumor cell lines with high procaspase-3 expression and in vivo xenografts. Direct activation of procaspase-3 can kill tumor cells directly without being affected by abnormalities in upstream apoptosis signaling pathways. Further development of more efficient and selective procaspase-3 activators as potential drugs is of great significance for tumor treatment. Summary of the Invention

[0004] This invention, based on the perspective of procaspase-3 activators, establishes a predictive model for activators targeting this target. It screens and designs approximately 1000 compounds from a series of natural products. Through extensive research, it optimizes and synthesizes a series of novel small molecule compounds with structures as shown in Formula I. Surprisingly, these compounds exhibit high antitumor activity, good selectivity, and low toxicity. These compounds, or their pharmaceutically acceptable salts, stereoisomers, tautomers, polymorphs, solvates, racemates, prodrugs, or metabolites, and their pharmaceutical compositions, can be used to treat and / or prevent cancers, such as lung cancer.

[0005] Therefore, the present invention provides a compound having the structure of Formula I or a pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, racemate, prodrug, or metabolite thereof:

[0006]

[0007] in,

[0008] R1, R2, R3, R4, and R5 are each independently selected from: -R6, -O-R6;

[0009] R6 is selected from: -H, -C 1-8 Alkyl, -C 2-8 Alkenyl; preferably, R6 is selected from: -C 3-6 Alkyl, -C 3-6 Alkenyl group.

[0010] The term "alkyl" refers to a saturated aliphatic hydrocarbon group, which can be a straight-chain or branched group.

[0011] The term "alkenyl" refers to an alkyl group as defined above that contains at least one carbon-carbon double bond, and can be a straight-chain or branched group.

[0012] Preferably, in Formula I, at least one of R1, R2, R3, R4, and R5 is selected from: -R6, -O-R6; wherein R6 is selected from: -C 3-6 Alkyl, -C 3-6 Alkenyl group.

[0013] Preferably, in Formula I, when R5 is -OH, R1, R2, R3 and R4 are not simultaneously -H; or, when R3 is -OH, R1, R2, R4 and R5 are not simultaneously -H; or, when R3 is -OH, R4 is not -O-CH3.

[0014] Preferably, in formula I, R5 is -OH; R4 is -C 3-8 Alkenyl group, preferably -C 3-6 Alkenyl group, more preferably -CH2CH=CH2.

[0015] Preferably, in formula I,

[0016] R3 is -OC 5-8 Alkyl or -OC 5-8 Alkenyl, preferably -O-CH2CH=C(CH3)2 or -O-hexyl; or,

[0017] R3 represents -OH or -OC. 5-8 Alkyl or -OC 5-8 Alkenyl group; R2 is -C 1-8 Alkyl, -OC 1-8 Alkyl, -C 2-8 alkenyl, -OC 2-8 alkenyl, more preferably -OC 3-8 Alkyl or -OC 3-8 Alkenyl group; R4 is -H.

[0018] Preferably, in formula I,

[0019] R4 is -O-CH3; R3 is -OC 3-8 Alkyl or -OC 3-8 Alkenyl group, preferably -O-CH2CH=C(CH3)2; or,

[0020] R4 is -O-CH3; R3 is -OH; R2 is -C 2-8 Alkenyl group, preferably -C 3-6 Alkenyl group, more preferably -CH2CH=CH2 or -CH2CH=C(CH3)2, and even more preferably -CH2CH=C(CH3)2.

[0021] In some specific embodiments, the compounds of Formula I and their pharmaceutically acceptable salts, stereoisomers, tautomers, polymorphs, solvates, racemates, prodrugs, or metabolites described in this invention are selected from:

[0022]

[0023]

[0024]

[0025] Preferably, the compound of formula I is selected from:

[0026]

[0027] This invention also provides a method for preparing a compound of formula I or a pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, racemic mixture, prodrug, or metabolite thereof, comprising the following steps:

[0028]

[0029] Compound A reacts with hydrazine hydrate to give compound B, and compound B undergoes a condensation reaction with compound C to give compound I. The definitions of R1, R2, R3, R4, and R5 are the same as described above.

[0030] Compound A is commercially available. Compound C is obtained from commercially available benzaldehyde compounds via substitution and optional rearrangement reactions.

[0031] In another aspect, the present invention also provides a pharmaceutical composition comprising at least one compound of formula I of the present invention or a pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, racemate, prodrug, or metabolite thereof, and one or more pharmaceutically acceptable carriers.

[0032] In another aspect, the present invention proposes the use of a compound of Formula I or a pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, racemic mixture, prodrug, or metabolite thereof, or a pharmaceutical composition thereof, in a medicament for the prevention and / or treatment of cancer. Preferably, the cancer is lung cancer.

[0033] The compound of Formula I of this invention exhibits strong tumor cell inhibitory activity, while demonstrating significant selectivity, low cytotoxicity towards normal cells, and low toxicity. This compound possesses antitumor activity, low toxicity, high selectivity, a mild and simple synthesis method, and is suitable for industrial production. Attached Figure Description

[0034] Figure 1 The values ​​represent the inhibition rates of the compounds of this invention against A549 cells. Note: The concentrations of each compound, from bottom to top along the Y-axis, are 10 μM, 30 μM, and 80 μM.

[0035] Figure 2 This is a prediction diagram of the blood-brain barrier model and intestinal absorption model for the compounds of this invention, namely, the diagrams of ADMET-PSA-2D and AlogP98. Note: Ellipses corresponding to the 95% and 99% confidence limits of the blood-brain barrier permeability (BBB) ​​and intestinal absorption models for the compounds of this invention. Detailed Implementation

[0036] The present invention will be further illustrated below with reference to specific embodiments. It should be understood that these embodiments are for illustrative purposes only and are not intended to limit the scope of the invention. Unless otherwise specified in the following embodiments, test methods performed under conventional conditions for such reactions are generally performed. The terminology used in this invention is for descriptive purposes only and not for limiting specific embodiments. Unless otherwise specified in the context, all technical and scientific terms used in this invention are intended to have the same meaning as commonly understood by those skilled in the art. The technical intent used in this invention refers to technology commonly understood in the art, including variations or equivalent substitutions of technology that are obvious to those skilled in the art.

[0037] Unless otherwise specified, all experimental materials and reagents used in the following examples are available from commercially available sources.

[0038] The structures of the compounds described in the following examples were determined by nuclear magnetic resonance. 1 H-NMR and 13 The determination was made by C-NMR. The NMR spectrometer used was an Agilent DD2 600MH NMR spectrometer. The solvents used for determination were deuterated methanol (CD3OD), deuterated chloroform (CDCl3), and hexadeuterated dimethyl sulfoxide (DMSO-d6). The internal standard was tetramethylsilane (TMS).

[0039] The abbreviations used in the nuclear magnetic resonance (NMR) data in the following examples have the following meanings:

[0040] s: singlet, d: doublet, t: triplet, q: quartet, dd: double doublet, qd: quartet doublet, ddd: double double doublet, ddt: double double triplet, dddd: double double double doublet, m: multiplet, br: broad, J: coupling constant, Hz: Hertz, δ: chemical shift, chemical shift (δ) values ​​are given in parts per million (ppm).

[0041] Example 1: Preparation of Compound 6

[0042]

[0043] Step 1: Synthesis of Compound 6-1

[0044] 3.84 g (19.77 mmol, 1.0 eq) of compound SM-1 was weighed into a reaction flask and dissolved in 40 mL of acetonitrile with stirring. 7.17 g (37.40 mmol, 1.9 eq) of EDCI and 8.55 g (63.27 mmol, 3.2 eq) of HOBT were added. After stirring at room temperature for 30 min, 6.5 mL (113.7 mmol, 5.7 eq) of 85% hydrazine hydrate was added, and the reaction was allowed to proceed for 4 h. The reaction progress was monitored by TLC. When the starting material disappeared, the reaction solution was cooled to -20 °C overnight, and the layers separated. The upper layer was poured off, 60 mL of water was added, and the mixture was stirred to dissolve. The solid precipitated by standing, filtered, and dried to obtain 3.50 g of white solid, with a yield of 85.02%. ESI-MS (m / z): 209.09 [M+H]+.

[0045] Step 2: Synthesis of Compound 6

[0046] 253.18 mg (1.22 mmol, 1.0 eq) of compound 6-1 was weighed into a reaction flask, 2.5 mL of anhydrous ethanol was added, and the mixture was stirred to dissolve. 0.25 g (1.54 mmol, 1.3 eq) of compound SM-2 (2-hydroxy-3-allylbenzaldehyde) was added, and the mixture was heated to 50 °C and reacted for 4 h. The reaction progress was monitored by TLC, and the starting material disappeared. The reaction solution was allowed to stand and cool, and a solid precipitated. The solid was filtered, and the filter cake was dried at 50 °C to give 0.22 g of a white solid, with a yield of 51.36%, HPLC purity of 98.29%, and ESI-MS (m / z): 353.15 [M+H]. + .

[0047] 1 H NMR(600MHz,DMSO-d6)δ11.99(s,1H),11.94(s,1H),9.58(s,1H),8.37(s,1H),7. 59(d,J=15.6Hz,1H),7.32(d,J=7.6Hz,1H),7.23(s,1H),7.18(d,J=7.4Hz,1H),7. 11(d,J=8.1Hz,1H),6.89(t,J=7.5Hz,1H),6.84(d,J=8.1Hz,1H),6.53(d,J=15.7 Hz,1H),6.06–5.92(m,1H),5.14–4.98(m,2H),3.84(s,3H),3.39(d,J=6.7Hz,2H).

[0048] 13C NMR(150MHz,DMSO-d6)δ162.16,155.93,149.42,148.85,148.36,142.20,137.06,131.94,12 9.55,127.54,126.48,122.63,119.50,118.02,116.34,116.16,111.56,56.01,33.82,19.03.

[0049] Example 2 Preparation of Compound 12

[0050]

[0051] Step 1: Synthesis of Compound 12-1

[0052] The synthesis steps are the same as those for compound 6-1.

[0053] Step 2: Synthesis of Compound 12-2

[0054] Weigh 1.50 g (9.86 mmol, 1.0 eq) of compound SM-2, add 20 ml of acetonitrile, weigh 2.72 g (19.68 mmol, 2.0 eq) of potassium carbonate, and weigh 2.20 g (14.76 mmol, 1.5 eq) of isopentenyl bromide. Reflux for 1.5 h, monitor the reaction progress by TLC, and the starting material disappears. Filter and wash the filter cake 2-3 times with acetonitrile, 5-10 ml each time. Collect the filtrate and concentrate under reduced pressure to obtain 2.40 g of brown oily substance.

[0055] Step 3: Synthesis of Compound 12

[0056] 179.84 mg (0.86 mmol, 1.0 eq) of compound 12-1 was weighed into a reaction flask, 2.5 mL of anhydrous ethanol was added, and the mixture was stirred to dissolve. Then, 0.19 g (0.86 mmol, 1.0 eq) of compound 12-2 (3-methoxy-4-O-isopentenylbenzaldehyde) was added, and the mixture was heated to 50 °C and reacted for 4 h. The reaction progress was monitored by TLC, and the starting material disappeared. The reaction solution was allowed to stand and cool, 2 mL of n-hexane was added, and the mixture was sonicated to precipitate a solid. The solid was filtered and dried in a 50 °C oven to obtain 0.24 g of a pale yellow solid, with a yield of 68.07%, HPLC purity of 97.32%, and ESI-MS (m / z): 411.19 [M+H]. + .

[0057] 1H NMR(600MHz,MeOD)δ8.04(s,1H),7.68(d,J=1.9Hz,1H),7.66(d,J=15.6Hz,1H ),7.16(d,J=2.0Hz,1H),7.13(dd,J=8.3,1.9Hz,1H),7.09(dd,J=8.2,2.0Hz, 1H),6.94(d,J=8.3Hz,1H),6.82(d,J=8.1Hz,1H),6.47(d,J=15.6Hz,1H),5.4 9–5.46(m,1H),4.58(d,J=6.8Hz,2H),3.90(s,6H),1.78(s,3H),1.75(s,3H).

[0058] 13 C NMR(150MHz,MeOD)δ165.70,152.00,151.35,150.35,149.35,144.05,139.18,128.64,128.13,124 .06,123.54,120.84,116.58,116.33,113.94,111.77,110.06,66.77,56.42,56.40,25.87,18.20.

[0059] The inventors also synthesized the following compounds using a similar synthesis method to that of Examples 1 and 2.

[0060] Table 1. Structural formulas and data analysis of the compounds of this invention.

[0061]

[0062]

[0063]

[0064]

[0065]

[0066]

[0067]

[0068] Example 3 In vitro activity study

[0069] The inhibitory activity of the compounds of this invention on human non-small cell lung cancer cells (A549) was investigated.

[0070] (1) Test method: The MTT assay was used to study the inhibitory effect of the compound on cancer cell proliferation.

[0071] (2) Instruments and reagents: see Table 2 and Table 3.

[0072] Table 2 Reagent List

[0073]

[0074] Table 3. List of Instruments

[0075]

[0076] (3) Compound preparation

[0077] The compound was dissolved in DSMO or anhydrous ethanol and stored at -20°C for later use. The sample in DSMO or anhydrous ethanol was then serially diluted with culture medium to the required concentration.

[0078] (4) Test methods

[0079] The specific steps are as follows: human non-small cell lung cancer cells (A549) are cultured in DMEM medium containing 10% PBS (penicillin and streptomycin are both 100kU / L) in a 37°C constant temperature incubator with 5% CO2. When the cells grow to 80-90% confluence, they are passaged, and the medium is replaced with fresh medium every 24 hours.

[0080] A549 cells were cultured at 37°C in a 5% CO2 incubator until the logarithmic growth phase. Cells were seeded into 96-well plates at a density of 5000 cells / well. After 24 hours, the plates were divided into a blank control group (NC), a positive control group (10, 30, 50 μM), and a test compound group (10, 30, 50 μM). After another 24 hours of culture, 10 μL of CCK-8 solution was added to each well. The 96-well plates were then returned to a 37°C, 5% CO2 incubator for another 2 hours. The absorbance (OD) of each well was measured at 450 nm using a microplate reader, and the cell inhibition rate was calculated.

[0081] Inhibition rate = [(Ac-As) / (Ac-Ab)] × 100%

[0082] As: Absorbance of experimental wells (including cells, culture medium, CCK-8 solution, and compound solution);

[0083] Ac: Absorbance of control wells (including cells, culture medium, and CCK-8 solution, excluding compound solutions);

[0084] Ab: Absorbance of blank wells (containing culture medium and CCK-8 solution, but excluding cells and compounds).

[0085] IC50 targeting compounds with strong cell-inhibiting effects 50Value detection. A549 cells were cultured in a 37℃, 5% CO2 incubator until the logarithmic growth phase, and then cultured at a cell density of 5×10⁻⁶. 4 100 μL of each compound was seeded at a concentration of 1 / mL into a 96-well plate. After 24 h, the plates were divided into a blank control group (NC) and test compound groups (0.1, 1, 5, 10, 30, and 80 μM). After further incubation for 24 h, the absorbance (OD) of each well was measured at 450 nm using a microplate reader, and the IC50 of each effective compound was calculated. 50 value.

[0086] In this embodiment, the half-maximal inhibitory activity of the compound of the present invention against cancer cells was determined, as shown in Table 4 below.

[0087] Table 4. Data on the inhibitory activity of the compounds of this invention against cancer cells.

[0088]

[0089] The results in Table 4 show that, in the study of A549 cell inhibition, the introduction of other groups (such as allyloxy, isopentenyloxy, and hexyloxy) at position 14 of the compounds of Formula II decreased the inhibitory activity against A549 cells. Conversely, the introduction of an allyl group at position 15 improved the inhibitory activity caused by the introduction of other groups at position 14. Therefore, it can be determined that the hydroxyl group at position 14 is the key group for inhibiting A549 cell proliferation. The introduction of an allyl group at position 15 enhances its inhibitory activity against cell proliferation. The presence of both an allyl group at position 15 and a hydroxyl group at position 14, similar to PAC-1, indicates good inhibitory activity against tumor cell proliferation.

[0090]

[0091] Introducing an allyloxy group at the 16-position of compound III did not improve its inhibitory activity against A549 cell proliferation, while introducing isopentenyloxy or hexyloxy significantly enhanced its inhibitory activity against A549 cells. Furthermore, introducing allyl, isopentenyl, or hexyl groups at the 15- or 17-positions improved its inhibitory activity. Retaining the 16-position hydroxyl group and introducing allyl, isopentenyl, or hexyl groups at the 15- or 17-positions also improved its inhibitory activity, but introducing allyl groups at both the 15- and 17-positions did not significantly improve its activity. Therefore, appropriate modifications to the structure of compound III are necessary and beneficial for inhibiting A549 cell proliferation.

[0092]

[0093] Introducing allyloxy, isopentenyloxy, and hexyloxy groups at the 16-position of compound IV enhances its inhibitory effect on A549 cell proliferation, with the introduction of isopentenyloxy showing the strongest inhibitory effect. However, introducing an allyl group at the 17-position does not show any improvement in inhibitory activity. Retaining the hydroxyl group at 16 and introducing an allyl or isopentenyl group at 17 significantly enhances the inhibitory activity, with the introduction of isopentenyl showing a more significant enhancement than that of allyl. Therefore, appropriate modifications to the structure of formula IV are necessary and beneficial for inhibiting A549 cell proliferation, and retaining the hydroxyl group at 16 and introducing a group at 17 further enhances the anti-cell proliferation effect.

[0094]

[0095] It is evident that the compounds of this invention have the effect of inhibiting the proliferation of cancer cells. Among them, except for compounds 1, 5, 14, 19, 21 and 23, all showed good inhibitory effects on cell viability, and were superior to the positive control drug PAC-1. In particular, compounds 4, 6, 12 and 20 had more obvious effects, among which compounds 6 and 12 had the best anti-lung cancer tumor effects.

[0096] Example 4: Study on the Prediction of ADMET Properties

[0097] The ADMET properties of the compounds of this invention were studied using Discovery Studio 2020 software.

[0098] ADMET properties refer to the absorption, distribution, metabolism, excretion, and toxicity of molecules within an organism. Predicting and selectively optimizing compounds based on their ADMET properties in the early stages of drug development is necessary and indispensable for improving the success rate of drug development and minimizing financial waste caused by ADMET properties in later stages.

[0099] The compounds of this invention were processed as small molecule compounds using the Prepare ligands, ADMETDescriptors, and Toxicity Predication modules in Discovery Studio 2020 software for pharmacokinetic and toxicity prediction. In the small molecule processing, Change Ionizatin, Generate Tautomers, and Generate Isomers were set to False, with other parameters left as default. All ADMET property prediction parameters were set to default. The toxicity prediction model was set to Rat Oral LD. 50 Other parameters are default.

[0100] In this embodiment, the ADMET properties of the compounds of the present invention were determined, as shown in Table 5 below.

[0101] Table 5 Results of the property prediction study of the compound ADMET of the present invention

[0102]

[0103]

[0104] As can be seen, all compounds of this invention have a certain degree of solubility. Except for compounds 1, 3, 5, 6, 11, 13, 19, 21, and 23, which have moderate blood-brain barrier permeability, the other compounds have poor permeability. It is presumed that poorly permeable compounds will not cause damage to the central nervous system. Furthermore, compared to the PAC-1 project, most compounds are safe for the central nervous system. None of the predicted compounds are CYP 2D6 inhibitors; therefore, it is presumed that these compounds will not cause side effects such as liver dysfunction. Compounds 4, 9, 14, and 24 are predicted to have hepatotoxicity, while compounds 9 and 24, after modification, do not have hepatotoxicity. Except for compound 29, all other compounds have good intestinal absorption levels. All predicted compounds can bind to plasma proteins. The compounds of this invention have lower oral toxicity compared to PAC-1.

[0105] Based on the above description of the invention, those skilled in the art can fully apply the present invention, and all modifications based on the same principles or similar modifications should be considered to be included within the scope of the present invention.

Claims

1. The following compounds or their pharmaceutically acceptable salts: 、 、 or 。 2. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein, Selected from the following compounds or their pharmaceutically acceptable salts: or 。 3. A pharmaceutical composition comprising at least one compound of claim 1 or a pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable carriers.

4. Use of the compound of claim 1 or a pharmaceutically acceptable salt thereof or the pharmaceutical composition of claim 3 in the preparation of a medicament for the treatment of non-small cell lung cancer.

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