Caffeic acylhydrazone compounds and uses thereof

Abstract

The present application belongs to the field of pharmaceutical chemistry, and relates to caffeic acylhydrazone compounds and purposes thereof. Specifically, the present application provides a caffeic acylhydrazone compound of formula I or a pharmaceutically acceptable salt thereof, which exhibits good antitumor activity.

Description

Technical Field

[0001] This invention belongs to the field of medicinal chemistry and relates to a caffeoylhydrazone compound with antitumor activity, comprising a pharmaceutical composition thereof, 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. Breast cancer became the most common cancer worldwide in 2020, followed by lung cancer, which is the leading cause of cancer death globally. Colorectal cancer was the third most common cancer, with the second-highest mortality rate after 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 cancer. 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, or pharmaceutical compositions thereof, can be used to treat and / or prevent cancers such as lung cancer.

[0005] Therefore, the present invention provides a caffeic acid hydrazone compound having the structure of Formula I or a pharmaceutically acceptable salt 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, R5 is -OH; R4 is -C 3-8 Alkenyl group, preferably -C 3-6 Alkenyl group, more preferably -CH2CH=CH2.

[0014] Preferably, in formula I,

[0015] R3 is -OC 5-8 Alkyl or -OC 5-8 Alkenyl, preferably -O-CH2CH=C(CH3)2 or -O-hexyl; or,

[0016] 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.

[0017] Preferably, in formula I,

[0018] R4 is -O-CH3; R3 is -OC 3-8 Alkyl or -OC 3-8 Alkenyl group, preferably -O-CH2CH=C(CH3)2; or,

[0019] 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.

[0020] Preferably, the compound of formula I or a pharmaceutically acceptable salt thereof described in this invention is selected from the following compounds or pharmaceutically acceptable salts thereof:

[0021]

[0022]

[0023] More preferably, the compound of formula I or a pharmaceutically acceptable salt thereof described in this invention is selected from the following compounds or pharmaceutically acceptable salts thereof:

[0024]

[0025]

[0026] More preferably, the compound of formula I or a pharmaceutically acceptable salt thereof of the present invention is selected from the following compounds or pharmaceutically acceptable salts thereof:

[0027]

[0028] The present invention also provides a method for preparing a compound of formula I or a pharmaceutically acceptable salt thereof, comprising the following steps:

[0029]

[0030] 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.

[0031] Compound A is commercially available. Compound C is obtained from commercially available benzaldehyde compounds via substitution and optional rearrangement reactions.

[0032] 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 thereof, and one or more pharmaceutically acceptable carriers.

[0033] In another aspect, the present invention provides for the use of a compound of formula I or a pharmaceutically acceptable salt thereof, or the aforementioned pharmaceutical composition, in the preparation of a medicament for the prevention and / or treatment of cancer. Preferably, the cancer is lung cancer or melanoma.

[0034] 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 invention's compound possesses antitumor activity, low toxicity, high selectivity, a mild and simple synthesis method, and is suitable for industrial production. Attached Figure Description

[0035] Figure 1 The value represents the inhibition rate of the compound of this invention on A549 cells.

[0036] Figure 2 The value represents the inhibition rate of the compound of this invention on A375 cells. Detailed Implementation

[0037] 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.

[0038] Unless otherwise specified, all experimental materials and reagents used in the following examples are available from commercially available sources.

[0039] 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 600 MH 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).

[0040] The abbreviations used in the nuclear magnetic resonance (NMR) data in the following examples have the following meanings:

[0041] s: singlet, d: doublet, t: triplet, q: quartet, dd: doubledoublet, qd: quartet doublet, ddd: doubledouble 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).

[0042] Example 1: Preparation of Compound 5

[0043]

[0044] Step 1: Synthesis of Compound 5-1

[0045] 8.29 g (46.02 mmol, 1.0 eq) of caffeic acid (SM-1) was weighed into a reaction flask and dissolved in 90 mL of acetonitrile with stirring. 11.74 g (61.25 mmol, 1.3 eq) of EDCI and 12.30 g (91.02 mmol, 1.98 eq) of HOBt were added. The mixture was stirred at room temperature for 30 min, then 10 mL (174.89 mmol, 3.8 eq) of 85% hydrazine hydrate was added. The reaction was carried out at 5 °C for 6 h, and the reaction progress was monitored by TLC. When the starting material disappeared, the reaction solution was cooled to -20 °C overnight. After separation, the upper layer was poured off, 90 mL of water was added, and the mixture was stirred to dissolve. The solid precipitated by standing, filtered, and dried to obtain 4.83 g of white solid, with a yield of 54.10% and a purity of 95.04%.

[0046] Step 2: Synthesis of Compound 5

[0047] 124.16 mg (0.64 mmol, 1.0 eq) of compound 5-1 was weighed into a reaction flask, 2.5 mL of anhydrous ethanol was added, and the mixture was stirred to dissolve. 107.65 mg (0.66 mmol, 1.0 eq) of 2-hydroxy-3-allylbenzaldehyde (SM-2) was added, and the mixture was heated to 50 °C and reacted for 4 h. The reaction was monitored by TLC, and the reaction was terminated when the starting material disappeared. The reaction solution was allowed to stand and cool, crystals precipitated and filtered, and then dried in a 50 °C oven for 2 h to obtain 105.42 mg of a yellow solid, with a yield of 48.73% and a purity of 96.39%.

[0048] 1H NMR(600MHz,MeOD)δ8.30(s,1H),7.64(d,J=15.6Hz,1H),7.20(d,J=7.6Hz,2H),7.09(d,J=2.0Hz,1H),6.99(dd,J=8.1,2.1Hz,1H) ,6.89(t,J=7.5Hz,1H),6.82(d,J=8.1Hz,1H),6.45(d,J=15.6Hz,1H),6.10–5.97(m,1H),5.17–5.02(m,2H),3.45(d,J=6.7Hz,2H).

[0049] 13 C NMR(150MHz,MeOD)δ165.49,157.66,151.84,149.85,147.29,145.15,138.32,133.59,1 30.87,129.56,128.49,123.08,120.70,119.18,116.96,116.23,116.01,115.62,35.20.

[0050] Example 2: Preparation of compound 23

[0051]

[0052] Step 1: Synthesis of Compound 23-1

[0053] The synthesis steps are the same as those for compound 5-1.

[0054] Step 2: Synthesis of Compound 23

[0055] 204.95 mg (1.08 mmol, 1.0 eq) of compound 23-1 was weighed into a reaction flask, 2.5 mL of anhydrous ethanol was added, and the mixture was stirred to dissolve. 211.09 mg (1.11 mmol, 1.0 eq) of 3-isopentenyl-4-hydroxybenzaldehyde (SM-2) was added, and the mixture was heated to 50 °C and reacted for 4 h. The reaction was monitored by TLC, and the reaction ended when the starting material disappeared. The mixture was concentrated under reduced pressure and purified by column chromatography. After drying, 147.93 mg of a yellow solid was obtained, with a yield of 37.47% and a purity of 96.86%.

[0056] 1H NMR (600MHz, MeOD) δ8.04(s,1H),7.61(d,J=16.0Hz,1H),7.53(d,J=2.0Hz,1H),7.50(dd,J=8.3,2.2Hz,1H),7.08(d,J=1.9Hz,1H),6.98(dd, J=8.2,1.9Hz,1H),6.81(d,J=8.1Hz,2H),6.44(dd,J=15.6,1.4Hz,1H),5.37–5.34(m,1H),1.79(d,J=16.4Hz,2H),1.76(s,3H),1.75(s,3H).

[0057] 13 C NMR(150MHz,MeOD)δ165.77,159.16,150.31,149.34,146.96,144.13,133.33,130.59,130.13 ,128.24,126.90,123.77,122.61,116.65,116.14,115.29,58.47,29.38,26.12,18.50,18.01.

[0058] The inventors synthesized the following compounds using a synthesis method similar to that of Examples 1 and 2.

[0059] Table 1. Structural formulas and data analysis of the compounds of this invention.

[0060]

[0061]

[0062]

[0063]

[0064]

[0065]

[0066] Example 3: In vitro activity

[0067] The inhibitory activity of the compounds of this invention against human non-small cell lung cancer (A549) and melanoma (A375) was investigated.

[0068] (1) Test method: The CCK-8 method was used to study the inhibitory effect of the compound on cancer cell proliferation.

[0069] (2) Instruments and reagents: see Table 2 and Table 3.

[0070] Table 2 Reagent List

[0071]

[0072] Table 3. List of Instruments

[0073]

[0074] (3) Compound preparation

[0075] 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.

[0076] (4) Test methods

[0077] The specific steps are as follows: human non-small cell lung cancer cells (A549) and human malignant melanoma cells (A375) 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% of the plate size, they are passaged, and the medium is replaced with fresh medium every 24 hours.

[0078] A549 and A375 cells were cultured at 37°C in a 5% CO2 incubator until the logarithmic growth phase. They were then 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), positive control groups (10, 30, and 50 μM), and test compound groups (10, 30, and 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.

[0079] Inhibition rate = [(Ac-As) / (Ac-Ab)] × 100%

[0080] As: Absorbance of experimental wells (including cells, culture medium, CCK-8 solution, and compound solution);

[0081] Ac: Absorbance of control wells (including cells, culture medium, and CCK-8 solution, excluding compound solutions);

[0082] Ab: Absorbance of blank wells (containing culture medium and CCK-8 solution, but excluding cells and compounds).

[0083] IC50 targeting compounds with strong cell-inhibiting effects 50 Value detection. A549 and A375 cells were cultured at 37°C in a 5% CO2 incubator until the logarithmic growth phase, and then cultured at a cell density of 5 × 10⁻⁶ cells / year. 4100 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.

[0084] In this embodiment, the half-maximal inhibitory activity of the compounds of the present invention against cancer cells was determined, as shown in Tables 4-5 below. Figure 1-2 As shown. Figure 1 The value represents the inhibition rate of the compound of this invention on A549 cells. Figure 2 The value represents the inhibition rate of the compound of this invention on A375 cells.

[0085] Table 4. Inhibitory activity data of the compounds of the present invention against A549 cells.

[0086]

[0087] Note: "-" indicates that it was not measured.

[0088] Table 5. Inhibitory activity data of the compounds of the present invention against A375 cells.

[0089]

[0090] Note: "-" indicates that it was not measured.

[0091] From Table 4-5 and Figure 1-2 The results show that compounds 1-3, 5-7, 9-10, 12-13, 16-18, and 21-29 exhibit significant inhibitory activity against A549 cells. Among these, compounds 16, 23, 29, 22, and 18 show better effects (IC50). 50 The values ​​were 5.63 μM, 5.68 μM, 6.96 μM, 7.20 μM, and 7.49 μM, respectively. Compound 16 showed better performance, with an IC50 value of 5.63 μM, 5.68 μM, 6.96 μM, 7.20 μM, and 7.49 μM. 50 The value is 5.63 μM.

[0092] Compounds 1, 5-7, 10, 13, 16-18, 20, 25, and 27-29 showed significant inhibitory activity against A375 cells. Among them, compounds 5, 10, 20, and 28 exhibited the best inhibitory effects (IC50, IC50, etc.). 50 The values ​​were 6.74 μM, 8.00 μM, 7.20 μM, and 8.15 μM, respectively, all lower than the IC of PAC-1. 50 (9.87 μM). Compound 5 showed better results, with an IC50 of 9.87 μM. 50 The value is 6.74 μM.

[0093] The technical intent used in this invention refers to the technology commonly understood in the art, including any variations or equivalent substitutions of the technology that are obvious to those skilled in the art.

Claims

1. The following caffeic acid hydrazone compounds or their pharmaceutically acceptable salts:

2. The caffeoylhydrazone compound according to claim 1, or a pharmaceutically acceptable salt thereof, wherein, The caffeoylhydrazone compound or a pharmaceutically acceptable salt thereof is selected from the following compounds or pharmaceutically acceptable salts thereof:

3. A pharmaceutical composition comprising at least one caffeoylhydrazone compound or a pharmaceutically acceptable salt thereof as described in any one of claims 1-2, and one or more pharmaceutically acceptable carriers.

4. Use of the caffeoylhydrazone compound of any one of claims 1-2 or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of claim 3, in the preparation of a medicament for the prevention and / or treatment of cancer.

5. The use according to claim 4, wherein, The cancer in question is either lung cancer or melanoma.

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