A naphthylphenyl ether compound, preparation method and application thereof

Abstract

The present invention provides a naphthylphenyl ether compound, a preparation method and application thereof, wherein the biologically active naphthylphenyl ether parent nucleus group contained therein is further chemically modified to produce a number of compounds with higher biological activity, thus expanding the wide application of such compounds in biomedicine and the development prospects of pharmaceutical preparations. Such compounds can significantly inhibit the proliferation of cells such as breast cancer, liver cancer, esophageal cancer and glioma, and inhibit the growth of tumor cells in the body and tumor formation, indicating that such compounds have the prospect of being developed into anti-tumor drugs.

Description

Technical Field

[0001] The present invention belongs to the field of tumor targeted therapy, and specifically relates to a naphthylphenyl ether compound, a preparation method and application thereof. Background Art

[0002] According to a report released by the International Agency for Research on Cancer in 2020, the global cancer burden continues to grow, with 18,094,716 new cases and 9,894,402 deaths from cancer in 2020. Among them, breast cancer patients have the highest new incidence rate, and lung cancer causes the most deaths. In addition, the incidence and mortality rates of colorectal cancer, gastric cancer, and liver cancer are also high. Traditional chemotherapy drugs such as paclitaxel, cisplatin, and doxorubicin can effectively inhibit tumor growth in the early stages, but after a period of treatment, drug resistance often occurs and the therapeutic effect is lost. The treatment response rate and efficacy of targeted anticancer drugs such as trastuzumab, gefitinib, and soratinib need to be improved. Therefore, there is an urgent need to develop new anticancer drugs.

[0003] STAT3 (Signal Transducer and Activator of Transcription 3) has been shown to be a key transcriptional regulator in tumor development. Continuous activation of STAT3 at tyrosine 705 is closely related to poor prognosis in breast cancer, liver cancer, and esophageal cancer. STAT3 protein can regulate the expression of a wide range of oncogenes, regulating malignant phenotypes such as proliferation, drug resistance, angiogenesis, and immune escape (Kamran MZ, Patil P, Gude RP. Role of STAT3 in cancer metastasis and translational advances. Biomed Res Int. 2013; 2013: 421821; Daniel E, Rachel A, Jennifer R. Targeting the IL-6 / JAK / STAT3 signalling axis in cancer. Nature Reviews Clinical Oncology, 2018, (6): 1-15.). STAT3 has played an important role in promoting the process of "inflammatory cancer transformation" in cells. Smoking, radiation, infection and drug stimulation can activate STAT3 signaling, promote the expression of pro-inflammatory cytokines such as IL-1β, IL-6, IL-8 and TNF-α, aggravate the inflammatory response of the local microenvironment, and communicate the core chain of inflammation-cancer transformation (Hua Yu, Drew Pardoll and Richard Jove. STATs in cancer inflammation and immunity: a leading role for STAT3. Cancer. 2009 (9): 798-809.). The development of drugs targeting STAT3 has become a hot topic of research at home and abroad.A variety of STAT3 inhibitors have entered the clinical testing stage (Hong, D., et al., AZD9150, a next-generation antisense oligonucleotide inhibitor of STAT3 with early evidence of clinical activity in lymphoma and lung cancer. Sci Transl Med, 2015, 7 (314): 314ra185; Ogura, M., et al., Phase I study of OPB-51602, an oral inhibitor of signal transducer and activator of transcription 3, in patients with relapsed / refractory hematological malignancies. Cancer Sci, 2015. 106 (7): 896-901.). However, these compounds are often limited in development due to their lack of efficacy, high toxicity or poor drugability. No drug targeting STAT3 has been successfully developed so far.

[0004] In view of the urgent need for clinical anticancer drugs and the lack of STAT3 inhibitors, the present invention aims to reveal the anticancer effects of a new class of naphthylphenyl ether compounds and their derivatives and their effects on STAT3 signals. Summary of the invention

[0005] The purpose of the present invention is to provide a naphthylphenyl ether compound, a preparation method and application thereof.

[0006] A naphthylphenyl ether compound, the structural formula of which is shown in general formula I:

[0007]

[0008] wherein R is selected from hydrogen, C1-C5 straight chain alkyl, C1-C5 straight chain alkyl substituted with halogen at the end, 5-8 membered cycloalkyl,

[0009] R1, R2, are each independently selected from H, F, Cl, Br, -CN, -CH3, -CF3, -OCH3, -OCF3,

[0010] E, G are carbon or nitrogen;

[0011] m represents the number of CH2 substituents, m is 0, 1, 2, 3;

[0012] n represents the number of CH2 substituents, n is 0, 1, 2, 3;

[0013] A is Where p represents the number of CH2 substituents, p is 1, 2, 3; Y is

[0014] Specifically, the compound has the following structure:

[0015]

[0016]

[0017] A biologically acceptable salt formed by the above naphthylphenyl ether compound and at least one of methanesulfonic acid, acetic acid, dihydrofolic acid, benzoic acid, citric acid, sorbic acid, propionic acid, oxalic acid, fumaric acid, maleic acid, hydrochloric acid, malic acid, phosphoric acid, sulfurous acid, sulfuric acid, vanillic acid, tartaric acid, ascorbic acid, boric acid, lactic acid and ethylenediaminetetraacetic acid.

[0018] The preparation method of the naphthylphenyl ether compound comprises the following steps:

[0019] (1) Potassium carbonate is dissolved in DMF, reacted at 100-150°C until complete, and then post-treated to obtain

[0020] (2) Potassium carbonate is dissolved in DMF and reacted at 100-150°C until complete. After post-treatment,

[0021] (3) Compound Dissolve in a mixed solution of ethanol and saturated ammonium chloride aqueous solution, add reducing agent iron powder under stirring at 40-50°C, react until complete, and obtain by column chromatography

[0022] (4) Dissolve triphosgene in DCM and add the compound dropwise at 0±5℃ and DIEA, react at room temperature for 1.5-2.5 hours, then add amine RNH2, add toluene, heat to 110-120°C for complete reaction, and obtain the target compound by column chromatography.

[0023] Furthermore, in step (1) The molar ratio with potassium carbonate is 1:3:3;

[0024] In step (2) The molar ratio of potassium carbonate is 1:1.2:2.2;

[0025] In step (3), The molar ratio of ethanol to iron powder is 1:5, and the volume ratio of ethanol to saturated ammonium chloride aqueous solution is 1:1;

[0026] In step (4) The molar ratio of triphosgene, DIEA, and amine RNH2 is 3:1:6:3.

[0027] The naphthylphenyl ether compounds and biologically acceptable salts thereof are used in the preparation of anti-tumor drugs, wherein the anti-tumor drugs refer to drugs for treating solid tumors, sarcomas, nervous system tumors and blood system cancers.

[0028] The use of the above-mentioned naphthylphenyl ether compounds and biologically acceptable salts thereof in the preparation of drugs for immune diseases related to JAKs-STAT3 cell signaling transduction, wherein the drugs for immune diseases related to JAKs-STAT3 cell signaling transduction refer to drugs for treating rheumatism, myelofibrosis, and psoriasis.

[0029] The naphthylphenyl ether compounds and biologically acceptable salts thereof are used in the preparation of drugs for treating immune reactions caused by bone marrow transplantation and immune factor storms caused by CAR-T immune cell therapy.

[0030] Another object of the present invention is to provide a class of small molecule compounds with anti-tumor activity.

[0031] The tumor may specifically be a tumor with high expression or constitutive activation of STAT3, including but not limited to breast cancer, liver cancer, esophageal cancer, glioma, etc.

[0032] Specifically, the present invention synthesized a class of naphthylphenyl ether compounds IY220924A-1 etc. with a completely new structure. The MTT method was used to detect the proliferation inhibition effect of the compounds on various cancer cells and the effect on the growth of subcutaneous transplanted tumors in mice.

[0033] The results show that the compound IY220924A-1 and the like of the present invention can effectively inhibit the proliferation and growth of breast cancer, liver cancer, esophageal cancer and glioma cells.

[0034] In summary, the present invention provides a new naphthylphenyl ether compound and its derivatives for use in tumor treatment and potential molecular mechanism. BRIEF DESCRIPTION OF THE DRAWINGS

[0035] Figure 1 To detect the effect of IY220924A-1 on the expression levels of target genes such as STAT3, p-STAT3, Cyclin D1, and C-myc in MDA-MB-468 cells by Western blot;

[0036] Figure 2The inhibitory effects of IY220924A-1 and IY220926A-1 on the growth of subcutaneous transplanted tumors in mice. DETAILED DESCRIPTION

[0037] In order to make the technical purpose, technical solution and beneficial effects of the present invention more clear, the technical solution of the present invention is further described below in conjunction with the accompanying drawings and specific embodiments.

[0038] In the method for synthesizing the compound of formula I of the present invention, various raw materials used in the reaction can be prepared by those skilled in the art according to prior knowledge, or can be prepared by methods known in the literature, or can be purchased commercially. The intermediates, raw materials, reagents, reaction conditions, etc. used in the above reaction schemes can be appropriately changed according to the prior knowledge of those skilled in the art.

[0039] In the present invention, unless otherwise specified: (i) the temperature is expressed in degrees Celsius (°C), and the operation is carried out at room temperature; more specifically, the room temperature refers to 20-30°C; (ii) the organic solvent is dried by a common drying method, and the solvent is evaporated using a rotary evaporator under reduced pressure, and the bath temperature is not higher than 50°C; the developing agent and the eluent are both in volume ratio; (iii) the reaction process is tracked by thin layer chromatography (TLC); (iv) the final product has a satisfactory proton nuclear magnetic resonance (1H-NMR).

[0040] Example 1: Synthesis of Compounds

[0041]

[0042] IY220924A-1: n=2,

[0043] IY220926A-1: n=2,

[0044] IY220925A-1: n=2,

[0045] IY220927A-1: n=2,

[0046] ID220925A-1: n=2,

[0047] ID220927A-1: n=2,

[0048] ID220926A-1: n=2,

[0049] ID220928A-1: n=2,

[0050] IY220927B-1: n=2,

[0051] IY220928A-1: n=2,

[0052] IY220929A-1: n=2,

[0053] IY220930A-1: n=2,

[0054] ID220928B-1: n=2,

[0055] ID220929A-1: n=2,

[0056] ID220930A-1: n=2,

[0057] ID220923A-1: n=2,

[0058] IY221001A-1: n=2,

[0059] IY221002A-1: n=2,

[0060] IY221003A-1: n=2,

[0061] IY221004A-1: n=2,

[0062] ID221001A-1: n=2,

[0063] ID221002A-1: n=2,

[0064] ID221003A-1: n=2,

[0065] ID221004A-1: n=2, IY221005A-1: n=2, IY221006A-1: n=2, ID221005A-1: n=2, ID221006A-1: n=2, The specific synthesis method takes compound IY220924A-1 as an example, and the structural formula is as follows:

[0066]

[0067] The name of compound IY220924A-1 is 1-(4-(4-(2-(piperidin-1-yl)ethoxy)phenoxy)naphthalen-1-yl)-3-(pyridin-4-ylmethyl)urea, and its synthesis route is as follows:

[0068]

[0069] Step 1. 4-((4-nitronaphthalen-1-yl)oxy)phenol (Compound 2)

[0070] 1-Fluoro-4-nitronaphthalene (compound 1, 2.0 g, 10.46 mmol, 1.0 eq), potassium carbonate (4.34 g, 31.39 mmol, 3.0 eq) and 1,4-benzenediol (compound 1a, 3.46 g, 31.39 mmol, 3.0 eq) were dissolved in 30 mL DMF and reacted at 120° C. for 16 hours. After the reaction was completed by TLC monitoring, potassium carbonate was filtered out, and DMF was removed by concentration under reduced pressure. Column chromatography (DCM / PE=1 / 5 to 1 / 1) was performed to obtain 2.5 g of 4-((4-nitronaphthalen-1-yl)oxy)phenol (compound 2) as a yellow solid with a yield of 85.0%.

[0071] Step 2. 1-(2-(4-((4-nitronaphthalen-1-yl)oxy)phenoxy)ethyl)piperidine (Compound 3)

[0072] Compound 2 (2.50 g, 8.89 mmol, 1.0 eq), compound 2a (1.96 g, 10.67 mmol, 1.2 eq), potassium carbonate (2.70 g, 19.55 mmol, 2.2 eq) were dissolved in 40 mL DMF and reacted at 120° C. for 16 hours. After the reaction was completed as monitored by TLC, potassium carbonate was filtered out, and DMF was removed by concentration under reduced pressure. Column chromatography (DCM / MeOH=1 / 0 to 40 / 1) was performed to obtain 3.12 g of 1-(2-(4-((4-nitronaphthalen-1-yl)oxy)phenoxy)ethyl)piperidine (compound 3), a brown oil, with a yield of 89.4%.

[0073] Step 3. 4-(4-(2-(piperidin-1-yl)ethoxy)phenoxy)naphthalen-1-amine (Compound 4)

[0074] Compound (3) (2.50 g, 6.37 mmol, 1.0 eq) was dissolved in 25 mL of ethanol and 25 mL of saturated aqueous ammonium chloride solution, the temperature was raised to 45°C, iron powder (1.78 g, 31.85 mmol, 5.0 eq) was slowly added in batches, and the temperature was raised to 55°C for reaction for 0.5 hours. After the reaction was completed by TLC monitoring, the mixture was filtered through diatomaceous earth, and the filtrate was extracted with dichloromethane 3 times (100 mL*3). The organic phases were combined, dried over anhydrous sodium sulfate, and dried by column (dichloromethane: methanol = 60:1 to 20:1) to obtain 1.42 g of 4-(4-(2-(piperidin-1-yl)ethoxy)phenoxy)naphthalen-1-amine (compound 4), a purple-red oil, with a yield of 61.5%.

[0075] Step 4.1-(4-(4-(2-(piperidin-1-yl)ethoxy)phenoxy)naphthalen-1-yl)-3-(pyridin-4-ylmethyl)urea(IY220924A-1)

[0076] Dissolve triphosgene (55 mg, 0.19 mmol, 1.0 eq) in DCM, cool to 0°C, slowly add compound 4 (201 mg, 0.56 mmol, 3.0 eq) and DIEA (144 mg, 1.1 mmol, 6.0 eq) dropwise, react at room temperature for 2 hours, then add compound 4a (60 mg, 0.56 mmol, 3.0 eq), add toluene and heat to 110°C for 12 hours. After the reaction is completed by TLC monitoring, directly spin dry and pass through a column (dichloromethane: methanol = 50:1 to 15:1) to obtain 65 mg 1-(4-(4-(2-(piperidin-1-yl)ethoxy)phenoxy)naphthalen-1-yl)-3-(pyridin-4-ylmethyl)urea (IY220924A-1), brown solid, yield 24.8%.

[0077] 1 H NMR(DMSO-d6,300MHz)δ:8.63(s,1H),8.53(d,J=6.0Hz,2H),8.11(d,J=9.0Hz,2H),7.74(d,J=9.0Hz,1H),7.58-7.50(m,2H),7.33 d,J=6.0Hz,2H),6.96-6.89(m,5H),4.34(d,J=4.0Hz,2H),4.11(m,2H),2.52(m,2H),1.53(m,4H),1.40(m,2H),1.39-1.20(m,2H).

[0078] The synthesis methods of other compounds refer to Example 1, except that the corresponding alkyl chloride is used in step 2, and the corresponding amine is used in step 4.

[0079] Example 2: Inhibitory effects of IY220924A-1 and its derivatives on proliferation of breast cancer, liver cancer, esophageal cancer and glioma cells

[0080] MDA-MB-468, BT-549, SUM159PT, HepG2, MHCC97H, HuH-7, SK-Hep1, KYSE-450, KYSE-30 and U251 cells in the logarithmic growth phase were collected and counted, and the cell suspension concentration was adjusted to 5 × 10 4 / mL, added to 96-well cell culture plates, 100μl per well. DMSO was used as solvent control, WP1066 (Chinese name: (2E)-3-(6-bromo-2-pyridinyl)-2-cyano-N-[(1S)-1-phenylethyl]-2-acrylamide, CAS: 857064-38-1, structure ) was used as a positive control. The naphthurea phenyl ether compound IY220924A-1 and its derivatives described in the present invention were diluted with DMSO and added to the culture wells, so that the final concentrations of the compounds in the system were 0.1, 0.3, 1, 3, 10, 30, 100 and 300 (μmol / L), respectively. After continuing to culture for 48 hours, 10 μL of MTT solvent (5 mg / ml) was added to each well, incubated at 37°C for 4 hours, the culture supernatant was discarded, 150 μl of DMSO was added to each well, and the cells were shaken and decolorized for 10 minutes. The OD value at an absorption wavelength of 490 nm was measured with an enzyme marker, and the results were recorded. The cell growth curve was drawn with the dosage of the compound as the horizontal axis and the absorbance value as the vertical axis. The half inhibition rate (IC 50 The statistical results of the values ​​are shown in the following table:

[0081] Table 1. MTT results of IY220924A-1 and its derivatives on breast cancer, liver cancer, esophageal cancer and glioma cells

[0082]

[0083]

[0084]

[0085]

[0086]

[0087]

[0088]

[0089]

[0090] As shown in Table 1, IY220924A-1 and its derivatives IY220926A-1, IY220925A-1 and IY220927A-1 have good proliferation inhibition effects on breast cancer, liver cancer, esophageal cancer and glioma cells. In particular, IY220924A-1 has stronger tumor inhibition activity in breast cancer and liver cancer cells. We focused on further studying the anti-tumor effect of IY220924A-1.

[0091] Example 3: Effect of IY220924A-1 on STAT3 signaling

[0092] MDA-MB-468 cells in the logarithmic growth phase were obtained and digested with trypsin. The cells were then cultured at a density of 3 × 10 5Single cell suspension of 100 μg / mL was prepared. 2 ml of cell suspension was inoculated into small dishes per well. The cells were incubated at 37°C and 5% CO2 overnight, and IY220924A-1 was added at concentrations of 0, 0.25 μM, 0.5 μM, 1 μM, and 2 μM. After 8 hours of continuous culture, the cells were lysed with RIPA lysis buffer, and the proteins were collected, quantified, and analyzed by Western blot.

[0093] like Figure 1 The results showed that as the concentration of IY220924A-1 increased, the protein expression of GAPDH and T-STAT3 remained unchanged, while the expression of p-STAT3 (Tyr705), Cyclin D1 and C-MYC showed a gradient downward trend. Therefore, IY220924A-1 can inhibit the activation of STAT3 and the protein expression of downstream target genes related to cell proliferation.

[0094] Example 4: Tumor-suppressing effect of IY220924A-1 in animals

[0095] The experimental animals were 5-week-old female Balb / c-Nude mice purchased from Beijing Weitonglihua Biotechnology Co., Ltd. and kept in an SPF animal laboratory. 6 MDA-MB-468 cells were inoculated in the armpits of mice. After about 10 days, obvious tumor nodules were formed in the armpits of mice. The mice were randomly divided into a solvent blank control group (blank suspension), an IY220924A-1 (1 mg / kg, 2.5 mg / kg, 5 mg / kg and 10 mg / kg) group and an IY220926A-1 (5 mg / kg) group, with 10 mice in each group. The drug was administered orally twice a day. The body weight and tumor size of mice in each group were monitored every other day.

[0096] Figure 2 The results showed that compared with the blank control group, the tumor volume of the IY220924A-1 (10 mg / kg) group and the IY220926A-1 (5 mg / kg) group was smaller, and multiple tumors disappeared in the IY220924A-1 and IY220926A-1 administration groups. Therefore, IY220924A-1 and its derivatives can not only inhibit tumor growth, but also inhibit tumor formation.

[0097] The above results show that IY220924A-1 and its derivatives can significantly inhibit the proliferation of breast cancer, liver cancer, esophageal cancer and glioma cells, inhibit the activation of STAT3 and the expression of downstream target genes, and show good anti-cancer effects in animals.

[0098] According to the general approach of drug development (conventional anti-tumor in vitro screening first, then targeted research), the compounds of the present invention can be applied to cancer treatment drugs related to abnormal cell proliferation, and can be prepared as anti-tumor drugs by mixing with human-acceptable salts or with pharmaceutical carriers.

[0099] Finally, it should be noted that the above embodiments are only used to illustrate but not limit the technical solutions of the present invention, and any equivalent substitutions of the present invention and any modifications or partial substitutions that do not depart from the spirit and scope of the present invention should be included in the scope of protection of the claims of the present invention.

Claims

1. A naphthylphenyl ether compound, characterized in that: The structural formula is shown in general formula I: Among them, R is R1 and R2 are each independently selected from H, F, Cl, Br, -CN, -CH3, -CF3, -OCH3, -OCF3, E, G are carbon or nitrogen; m represents the number of CH2 substituents, m is 0, 1, 2, 3; n represents the number of CH2 substituents, n is 0, 1, 2, 3; A is Where p represents the number of CH2 substituents, p is 1, 2, 3; Y is 2. A naphthylphenyl ether compound, characterized in that: Specifically, the compound has the following structure:

3. A biologically acceptable salt formed by the naphthylphenyl ether compound according to claim 1 or 2 and at least one of methanesulfonic acid, acetic acid, dihydrofolic acid, benzoic acid, citric acid, sorbic acid, propionic acid, oxalic acid, fumaric acid, maleic acid, hydrochloric acid, malic acid, phosphoric acid, sulfurous acid, sulfuric acid, vanillic acid, tartaric acid, ascorbic acid, boric acid, lactic acid and ethylenediaminetetraacetic acid.

4. The method for preparing the naphthylphenyl ether compound according to claim 1, characterized in that: The following steps are involved: (1) Potassium carbonate is dissolved in DMF, reacted at 100-150°C until complete, and then post-treated to obtain (2) Potassium carbonate is dissolved in DMF and reacted at 100-150°C until complete. After post-treatment, (3) Compound Dissolve in a mixed solution of ethanol and saturated ammonium chloride aqueous solution, add reducing agent iron powder under stirring at 40-50°C, react until complete, and obtain by column chromatography (4) Dissolve triphosgene in DCM and add the compound dropwise at 0±5℃ and DIEA, react at room temperature for 1.5-2.5 hours, then add amine RNH2, add toluene, heat to 110-120°C for complete reaction, and obtain the target compound by column chromatography.

5. The method for preparing naphthylphenyl ether compounds according to claim 4, characterized in that: In the step (1) The molar ratio with potassium carbonate is 1:3:3; In step (2) The molar ratio of potassium carbonate is 1:1.2:2.2; In step (3), The molar ratio of ethanol to iron powder is 1:5, and the volume ratio of ethanol to saturated ammonium chloride aqueous solution is 1:1; In step (4) The molar ratio of triphosgene, DIEA, and amine RNH2 is 3:1:6:

3.

6. Use of the naphthylphenyl ether compound according to claim 1 or 2 or the biologically acceptable salt of the naphthylphenyl ether compound according to claim 3 in the preparation of an anti-tumor drug, characterized in that: The anti-tumor drug refers to a drug for treating breast cancer, liver cancer, esophageal cancer or glioma.

7. Use of the naphthylphenyl ether compound according to claim 1 or 2 or the biologically acceptable salt of the naphthylphenyl ether compound according to claim 3 in the preparation of a drug for immune diseases associated with JAKs-STAT3 cell signal transduction, characterized in that: The immune disease drug related to JAKs-STAT3 cell signal transduction refers to a drug for treating rheumatism, myelofibrosis, and psoriasis.

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