An antitumor azacyclic hydrazide compound

Abstract

The present application belongs to the field of pharmaceutical chemistry, and relates to an anti-tumor azacyclic hydrazide compound. Specifically, the present application provides an anti-tumor azacyclic hydrazide compound of a structure of formula I or a pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, racemate, prodrug or metabolite thereof, which exhibits good anti-tumor activity.

Description

TECHNICAL FIELD

[0001] The present application belongs to the field of pharmaceutical chemistry, and relates to an anti-tumor nitrogen heterocyclic hydrazide compound or a pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, racemate, prodrug or metabolite thereof, a pharmaceutical composition containing the same, and a use thereof. BACKGROUND

[0002] Cancer is a malignant disease with extremely high morbidity and mortality and relatively poor treatment effect worldwide. In the global scope, anti-tumor drugs have become the first treatment area in the drug market. The commonly used anti-tumor drugs currently include cytotoxic drugs, hormone drugs, molecular targeted therapy drugs, biological response modifiers, tumor differentiation inducers, tumor angiogenesis inhibitors, and drugs for assisting tumor treatment, etc.

[0003] According to the research of the World Health Organization, in 2020, there were 1929 million new cancer cases worldwide, of which 4573 million new cancer cases in China accounted for 23.7% of the world, ranking first, which is about twice the number of new cancer cases in the United States, which ranks second. In 2020, breast cancer has become the cancer with the highest incidence worldwide, and lung cancer ranks second, but lung cancer is the cancer with the highest mortality rate worldwide. The second is colorectal cancer, which ranks third in the number of cases, and the cancer mortality rate is only second to lung cancer. Therefore, the discovery and development of anti-tumor drugs have always been the focus of global scientists and the key issues of major health and survival problems faced by mankind.

[0004] At present, anti-tumor drugs have the characteristics of high toxicity and low selectivity. Therefore, it is of great significance to discover and develop new compounds with higher activity, better selectivity and lower toxicity. SUMMARY

[0005] Based on the scientific hypothesis that "cancer is a body ulcer" and "cancer treatment should be preceded by ulcer treatment", about 30000 compounds are designed and synthesized from a series of anti-inflammatory drugs or lead compounds. A series of novel small molecule compounds with structures as shown in Formula I are synthesized by a large number of researches and optimized screening. It is surprisingly found that they all have high anti-tumor 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 for treating or / and preventing cancer, such as lung cancer, breast cancer, colon cancer and melanoma.

[0006] The present application provides an anti-tumor nitrogen heterocyclic hydrazide compound having a structure of Formula I or a pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, racemate, prodrug or metabolite thereof:

[0007]

[0008] wherein,

[0009] R1, R2, R3, R4and R5are each independently selected from: -R8, -O-R8;

[0010] R6is selected from: -R8, and is optionally substituted with one or more halogen;

[0011] R7is selected from: -R8, halogen;

[0012] R8is selected from: -H, -C 1-8 alkyl, -C 2-8 alkenyl, -C 3-8 cycloalkyl, -C 3-8 cycloalkenyl.

[0013] The "alkyl" group refers to a saturated aliphatic hydrocarbon radical which can be straight chain or branched chain.

[0014] The "alkenyl" group refers to an alkyl radical as defined above containing at least one carbon-carbon double bond, which can be straight chain or branched chain.

[0015] The "cycloalkyl" group refers to a saturated, monocyclic or polycyclic cyclic hydrocarbon substituent.

[0016] The "cycloalkenyl" group refers to a partially unsaturated, monocyclic or polycyclic cyclic hydrocarbon substituent.

[0017] The "halogen" refers to fluorine, chlorine, bromine or iodine.

[0018] Preferably, in formula I, R6is selected from: -C 1-4 alkyl substituted with one or more halogen, and R6is para (4-) substituted; more preferably from: para (4-) trifluoromethyl (-CF3).

[0019] Preferably, in formula I, R7is selected from: halogen; more preferably from: -F, -Cl, -Br; further preferably from: -F.

[0020] Preferably, in formula I, R8is selected from: -H, -C 1-6 alkyl, -C 2-6 alkenyl, -C 3-6 cycloalkyl, -C 3-6 cycloalkenyl; more preferably from: -H, -C 1-6 alkyl, -C 2-6 alkenyl.

[0021] Preferably, in formula I, at least one of R1, R2, R3, R4and R5is selected from: -R8, -O-R8; said R8is selected from: -C 1-8 alkyl, -C 2-8 alkenyl, -C3-8 cycloalkyl, -C 3-8 cycloalkenyl; more preferably selected from the group consisting of: -C 1-6 alkyl, -C 2-6 alkenyl, -C 3-6 cycloalkyl, -C 3-6 cycloalkenyl.

[0022] Preferably, in Formula I, R1, R3, and R5 are each independently selected from the group consisting of: -H, -OH, -O-C 1-6 alkyl, -O-C 2-6 alkenyl.

[0023] Preferably, in Formula I, R2 and R4 are each independently selected from the group consisting of: -H, -O-C 1-3 alkyl, -C 2-6 alkenyl.

[0024] Preferably, in Formula I, R1, R3, and R5 are each independently selected from the group consisting of: -H, -OH, -O-CH3, -O-CH2CH=CH2, -O-CH2CH=C(CH3)2, -O-(CH2)5CH3, -O-(CH2)3CH3.

[0025] Preferably, in Formula I, R2 and R4 are each independently selected from the group consisting of: -H, -OCH3, -CH2CH=CH2, -CH2CH=C(CH3)2.

[0026] Preferably, in Formula I, R2 is selected from the group consisting of: -CH2CH=CH2, -CH2CH=C(CH3)2.

[0027] In certain embodiments, the compounds of Formula I, and pharmaceutically acceptable salts, stereoisomers, tautomers, polymorphs, solvates, racemates, prodrugs, or metabolites thereof, are selected from the group consisting of:

[0028]

[0029]

[0030] The present application also provides a method of making a compound of Formula I, or a pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, racemate, prodrug, or metabolite thereof, comprising the steps of:

[0031]

[0032] Compound A is reacted with alcohol to form ester, i.e. compound B; compound B is reacted with R6-substituted benzyl bromide to form compound C; compound C is reacted with hydrazine hydrate to form compound D, and compound D is condensed with compound E to form compound of formula I. In the formula, R1, R2, R3, R4, R5, R6, R7 and R8 are the same as defined above.

[0033] Compound A can be directly obtained from the market. Compound C is obtained from commercially available benzaldehyde compounds through substitution reaction and optional rearrangement reaction.

[0034] In another aspect of the present application, the present application further provides a pharmaceutical composition comprising at least one compound of formula I or a pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, racemate, prodrug or metabolite thereof, and one or more pharmaceutically acceptable carriers.

[0035] In another aspect of the present application, the present application provides the use of a compound of formula I or a pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, racemate, prodrug or metabolite thereof, or a pharmaceutical composition for preventing and / or treating cancer, wherein the cancer is mainly lung cancer, breast cancer, colon cancer, melanoma.

[0036] The compound of the present application has strong tumor cell inhibitory activity, and shows obvious selectivity, low killing effect on normal cells and low toxicity. The compound of the present application has anti-tumor activity, low toxicity, strong selectivity, mild synthesis method, simple operation, easy synthesis and suitability for industrial production. DETAILED DESCRIPTION

[0037] The present application will be further described in conjunction with specific examples. It should be understood that these examples are only used to illustrate the present application and not used to limit the scope of the present application. In the following examples, if the specific conditions are not indicated, the test method is generally in accordance with the conventional conditions of such reactions. The terms used in the present application are only used for description and not for limiting the specific embodiments. Unless otherwise specified in the context, the meanings of all technical terms and scientific terms used in the present application are intended to be the same as generally understood by those skilled in the art. The technology used in the present application is intended to refer to the technology generally understood in the art, including changes or replacement of equivalent technology which are obvious to those skilled in the art.

[0038] The experimental materials and reagents used in the following examples are commercially available unless otherwise specified.

[0039] The structures of the compounds described in the following examples are determined by nuclear magnetic resonance 1 H-NMR and 13C-NMR was determined. The determination instrument of nuclear magnetic resonance used Agilent DD2 600MH nuclear magnetic resonance spectrometer, the determination solvent was deuterated methanol (CD3OD), deuterated chloroform (CDCl3), hexadeuterated dimethyl sulfoxide (DMSO-d6), and the internal standard substance was tetramethylsilane (TMS).

[0040] The meaning of the abbreviations represented in the nuclear magnetic resonance (NMR) data in the following examples is as follows:

[0041] s: singlet, d: doublet, t: triplet, q: quartet, dd: double doublet, qd: quartet doublet, ddd: double double doublet, ddq: double double triplet, ddd: double double double doublet, m: multiplet, br: broad, J: coupling constant, Hz: hertz, δ: chemical shift, the chemical shift (δ) value is given in units of parts per million (ppm).

[0042] Preparation of compound 22 of example 1

[0043]

[0044] Step one: synthesis of intermediate 1

[0045] SM1 25.0 g was weighed, stirred and dissolved in anhydrous ethanol, 37.5 g of concentrated sulfuric acid was added dropwise, and the reaction was refluxed. After the reaction was completed, the reaction was cooled to room temperature, 50 ml of water was added, about 80 g of potassium carbonate was added in batches, stirred for 1 hour, the pH was controlled to be greater than 8, the ethanol was removed under reduced pressure, 150 ml of water was added, mixed uniformly, then 100 ml of ethyl acetate was added for extraction, the water phase was collected, 200 ml of dichloromethane was added for extraction, and the operation was repeated once. The organic phase was collected, dried over anhydrous sodium sulfate, filtered, and the filtrate was evaporated under reduced pressure to obtain a white solid of about 4.03 g, with a yield of 15.10%.

[0046] Step two: synthesis of intermediate 2

[0047] Intermediate 1 2.01 g was weighed, stirred and dissolved in dichloromethane 15 ml, 760.5 mg of triethylamine was added, 1.62 g of p-trifluoromethyl benzyl bromide was added, and the reaction was stirred at room temperature. After the reaction was completed, dichloromethane was added, washed with water twice, dried over anhydrous sodium sulfate, filtered, and the filtrate was evaporated under reduced pressure to obtain a white solid of about 2.11 g, with a yield of 68.20%.

[0048] Step three: synthesis of intermediate 3

[0049] Take intermediate 2 2.0 g, add 30 ml of anhydrous ethanol, stir and dissolve, add hydrazine hydrate 4 ml, reflux reaction, after the reaction is completed, reduce to room temperature, filter, wash with appropriate amount of ethanol, and dry to obtain 1.51 g of white solid, yield: 75.30%.

[0050] Step four: synthesis of compound 22

[0051] Take intermediate 3 (108.8 mg, 1.0 eq), 3-allyl-4-allyloxybenzaldehyde (87.5 mg, 1.5 eq), add appropriate amount of anhydrous ethanol, reflux reaction, after the reaction is completed, reduce to room temperature, filter and wash with anhydrous ethanol, and dry the filter cake to obtain 120.5 mg of solid, yield: 80.56%.

[0052] 1 H NMR (600 MHz, CDC13) δ 13.17 (s, 1H), 8.81 (s, 1H), 8.15 (s, 1H), 8.08 (d, J = 13.1 Hz, 1H), 7.70 (d, J = 2.2 Hz, 1H), 7.60 (d, J = 8.0 Hz, 2H), 7.56 (dd, J = 8.5, 2.2 Hz, 1H), 7.49 (d, J = 7.9 Hz, 2H), 6.85 (d, J = 8.5 Hz, 1H), 6.81 (d, J = 6.8 Hz, 1H), 6.09 - 5.96 (m, 2H), 5.46 - 5.26 (m, 2H), 5.12 - 5.01 (m, 2H), 4.58 (dt, J = 5.0, 1.6 Hz, 2H), 4.31 (q, J = 7.3 Hz, 2H), 3.65 (s, 2H), 3.44 (dd, J = 6.7, 1.6 Hz, 2H), 3.31 (t, J = 4.8 Hz, 4H), 2.68 (t, J = 4.8 Hz, 4H), 1.56 (t, J = 7.2 Hz, 3H).

[0053] 13C NMR (150 MHz, CDC13) δ 175.24 (175.23), 161.78, 158.27, 154.32 (152.67), 148.57, 147.08, 145.52 (145.45), 142.24, 136.61, 133.19, 129.85 (129.63), 129.55, 129.36, 129.28, 128.01, 126.78, 125.48 (125.46, 125.43, 125.40), 125.22 (123.42), 122.38 (122.33), 117.36, 115.85, 113.22 (113.07), 111.45, 111.06, 103.86, (103.84), 68.91, 62.45, 52.92, 50.16, 50.13, 49.50, 34.64, 14.61.

[0054] Synthesis of compound 27 of example 2

[0055]

[0056] Step one: synthesis of intermediate 1

[0057] The synthesis step of intermediate 1 is the same as in example 1.

[0058] Step two: synthesis of intermediate 2

[0059] The synthesis step of intermediate 2 is the same as in example 1.

[0060] Step three: synthesis of intermediate 3

[0061] The synthesis step of intermediate 3 is the same as in example 1.

[0062] Step four: synthesis of compound 27

[0063] Intermediate 3 (108.1 mg, 1.0 eq), 4-hydroxybenzaldehyde (47.1 mg, 1.5 eq) were weighed, and an appropriate amount of anhydrous ethanol was added, and the reaction was refluxed. After the reaction was completed, it was naturally cooled to room temperature, filtered and washed with anhydrous ethanol, and oven-dried to obtain 109.1 mg of solid, with a yield of 83.28%.

[0064] 1H NMR (600 MHz, DMSO-d6) δ 13.13 (s, 1H), 9.91 (s, 1H), 8.88 (s, 1H), 8.27 (s, 1H), 7.91 (d, J = 13.3 Hz, 1H), 7.72 (d, J = 7.8 Hz, 2H), 7.59 (d, J = 8.1 Hz, 4H), 7.14 (d, J = 7.2 Hz, 1H), 6.83 (d, J = 8.2 Hz, 2H), 4.55 (q, J = 7.2 Hz, 2H), 3.67 (s, 2H), 3.30 (d, J = 5.6 Hz, 4H), 2.61 (t, J = 4.7 Hz, 4H), 1.40 (t, J = 7.1 Hz, 3H).

[0065] 13 C NMR (150 MHz, DMSO-d6) δ 173.91, 160.63, 159.34, 153.45 (151.81), 147.81, 147.58, 144.44, 143.07, 136.47, 129.40, 128.91, 125.32, 125.05, 125.03, 121.08, 115.59, 111.51 (111.36), 109.78, 105.71, 61.04, 52.21, 49.61, 48.55, 14.33..

[0066] The present inventors have also synthesized the following compounds using a synthetic method similar to that of Example 1 and Example 2.

[0067] Table 1 Structural formulae and data analysis of compounds of the present application

[0068]

[0069]

[0070]

[0071]

[0072]

[0073]

[0074]

[0075]

[0076]

[0077]

[0078]

[0079]

[0080]

[0081]

[0082]

[0083]

[0084]

[0085]

[0086] Example 3 In vitro activity study

[0087] The compounds of this invention exhibit inhibitory activity against human non-small cell lung cancer cells (A549), human breast cancer cells (MCF-7), human melanoma cells (A375), and human colon cancer cells (HCT-116).

[0088] 1. Testing method: The MTT assay was used to study the inhibitory effect of the compound on cancer cell proliferation.

[0089] 2. Instruments and reagents are listed in Tables 2 and 3.

[0090] Table 2 Reagent List

[0091]

[0092] Table 3. List of Instruments

[0093]

[0094] 3. Compound preparation

[0095] The compound was dissolved in DSMO and stored at room temperature under nitrogen atmosphere for later use. The sample in DSMO was then serially diluted with culture medium to the required concentration.

[0096] 4. Test methods

[0097] The specific steps are as follows: human non-small cell lung cancer cells (A549), human breast cancer cells (MCF-7), human melanoma cells (A375), and human colon cancer cells (HCT-116) are cultured in DMEM medium containing 10% FBS (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 on the plate, they are passaged, and the medium is replaced with fresh medium every 24 hours.

[0098] A549, MCF-7, HCT-116, A375 cells were cultured in a 37℃, 5% CO2 incubator to the logarithmic growth phase, and were inoculated in a 96-well plate at a cell density of 4000 cells / well. After 24 h, they were divided into a blank control group (NC), a positive drug group (2 μM), and a test compound group (2 μM). After 48 h of culture, 20 μL of MTT solution (5 mg·mL -1 ) was added to each well, and the culture was continued for 4 h. The culture solution was carefully aspirated, 150 μL of DMSO solution was added to each well, and oscillation was performed for 10 min to completely dissolve it. The absorbance value (OD) of each well was determined by an enzyme-labeled instrument at 490 nm, and the cell inhibition rate was calculated.

[0099] In this example, the inhibitory activity of the compounds of the present application on cancer cells was determined, as shown in Table 4 below.

[0100] Table 4 Data of the inhibitory activity of the compounds of the present application on cancer cells (48 h)

[0101]

[0102]

[0103] Note: The inhibition rate is negative.

[0104] As can be seen from the results in Table 4: in the study of the inhibitory activity on A549 cells, the compounds 1, 7, 17, 21 and 27 of the present application showed good inhibitory effects.

[0105] In the study of the inhibitory activity on MCF-7 cells, the compounds 1-3, 5-14, 17-22, 27 and 28 of the present application all showed good inhibitory effects on the activity of MCF-7 cells, especially the effects of compounds 18, 19, 21, 22, 27 and 28 were more obvious, and the anti-breast cancer effect of compound 27 was the best.

[0106] In the study of the inhibitory activity on HCT116 cells, the compounds 1, 6, 15, 17, 21, 26 and 27 of the present application had obvious inhibitory activity.

[0107] In the study of the inhibitory activity on A375 cells, among the compounds of the present application, compounds 1, 7, 11, 13, 17, 21 and 27 had obvious inhibitory activity.

[0108] Based on the foregoing description of the application, those skilled in the art can fully apply the application, and all identical principles or similar modifications should be considered as included in the scope of the application.

Claims

1. A compound of Formula I, or a pharmaceutically acceptable salt thereof, ###0001### Formula I wherein, R6 is selected from the group consisting of: para-trifluoromethyl; R1, R2, R3, R4and R5are each independently selected from the group consisting of: -R8, -O-R8; R8is selected from the group consisting of: -H, -C 1-8 alkyl, -C 2-8 alkenyl; R7 is selected from the group consisting of: -F.

2. The compound of Formula I according to claim 1, or a pharmaceutically acceptable salt thereof, wherein, 3. The compound of Formula I according to claim 1, or a pharmaceutically acceptable salt thereof, wherein, R8is selected from: -H, -C 1-6 alkyl, -C 2-6 alkenyl.

4. The compound of Formula I according to claim 3, or a pharmaceutically acceptable salt thereof, wherein, at least one of R1, R2, R3, R4and R5is selected from: -R8, -O-R8; R8is selected from: -C 1-8 alkyl, -C 2-8 alkenyl.

5. The compound of Formula I according to claim 1, or a pharmaceutically acceptable salt thereof, wherein, R8is selected from: -C 1-6 alkyl, -C 2-6 alkenyl.

6. The compound of Formula I according to claim 1, or a pharmaceutically acceptable salt thereof, wherein, R1, R3and R5are each independently selected from the group consisting of: -H, -OH, -O-C 1-6 alkyl, -O-C 2-6 alkenyl; R2and R4are each independently selected from the group consisting of: -H, -O-C 1-3 alkyl, -C 2-6 alkenyl. R1, R3 and R5 are each independently selected from the group consisting of: -H, -OH, -O-CH3, -O-CH2CH=CH2, -O-CH2CH=C(CH3)2, -O-(CH2)5CH3, -O-(CH2)3CH3; R2 and R4 are each independently selected from the group consisting of: -H, -OCH3, -CH2CH=CH2, -CH2CH=C(CH3)2.

7. The compound of Formula I according to claim 1, or a pharmaceutically acceptable salt thereof, wherein, R2 is selected from the group consisting of: -CH2CH=CH2, -CH2CH=C(CH3)2. The compound of Formula I is selected from the group consisting of:

8. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein 9. A pharmaceutical composition comprising at least one compound of Formula I according to any one of claims 1-8, or a pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable carriers.

10. Use of a compound of Formula I according to any one of claims 1-8, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 9, for the manufacture of a medicament for the prevention and / or treatment of cancer. The cancer is lung cancer, breast cancer, colon cancer or melanoma.

11. Use according to claim 10, wherein, ​

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