A mononitroisosorbide derivative and its preparation method and application

Abstract

The present invention relates to a mononitroisosorbide derivative, its preparation method, and application. The derivative has the following structure: #imgabs0#, wherein R is one of the following structures: #imgabs1#. The mononitroisosorbide derivative is prepared by reacting 5-isosorbide mononitrate with an acyl chloride (sulfonyl chloride) compound in the presence of triethylamine and a catalyst, 4-dimethylaminopyridine. The mononitroisosorbide derivative exhibits excellent anti-tumor activity. Compared to existing technologies, the present invention designs and synthesizes multiple compounds with novel structures, among which ZM533 exhibits moderate anti-tumor activity against the A549 cell line and is expected to be used in the preparation of anti-lung cancer drugs.

Description

Technical Field

[0001] The present invention relates to the technical field of drug synthesis, in particular to a mononitroisosorbide derivative and a preparation method and application thereof. Background Art

[0002] Targeted covalent modification has become an effective drug discovery approach. These drugs bind to their targets through irreversible or irreversible Michael addition reactions with unique cysteine residues, such as those on specific proteins. This strategy can enhance ligand binding selectivity for proteins within the same family, increase affinity for target proteins with shallow binding sites, and improve the success rate of drug discovery. The CD47-SIRPα pathway is one of the pathways that regulates the immune system and promotes the phagocytic function of phagocytes to achieve anti-tumor effects.

[0003] RRx-001, a small molecule inhibitor of the CD47-SIRPα pathway, covalently binds to cysteine residues on the CD47 target on phagocytes and has been shown to effectively activate autoimmunity. It is currently in Phase III clinical trials. RRx-001's chemical structure contains two nitro groups. Studies have suggested that one possible mechanism of RRx-001's anti-tumor activity is through its role as a nitric oxide donor. Literature research has shown that nitric oxide can regulate signaling pathways involved in tumor progression. High concentrations of nitric oxide can mediate macrophage phagocytosis of tumor cells, generate N / O free radicals within the cells, activate the expression of p53 and other proteins, and induce apoptosis. Isosorbide mononitrate is an effective drug for relieving angina pectoris, and its mechanism of action is as a nitric oxide donor. Therefore, using isosorbide mononitrate as a lead compound and introducing a covalently bound fragment is expected to yield new advances in the synthesis of anti-tumor drugs. Summary of the Invention

[0004] The purpose of the present invention is to provide a mononitroisosorbide derivative and its preparation method and application.

[0005] The purpose of the present invention can be achieved by the following technical solution: A mononitroisosorbide derivative has the following structure:

[0006]

[0007] Wherein, R is one of the following structures:

[0008]

[0009] Preferably, the R is the following structure:

[0010]

[0011] A method for preparing the above-mentioned mononitroisosorbide derivatives comprises reacting 5-isosorbide mononitrate with an acyl chloride or sulfonyl chloride compound in the presence of a base triethylamine (Et3N) and a catalyst 4-dimethylaminopyridine (DMAP) to produce the mononitroisosorbide derivatives ZM532-ZM536.

[0012]

[0013] (a)Acyl chloride, DCM, Et3N, DMAP

[0014] Preferably, the acyl chloride or sulfonyl chloride compound includes aminosulfonyl chloride, vinylsulfonyl chloride, p-toluenesulfonyl chloride, acryloyl chloride, and methacryloyl chloride.

[0015] Preferably, the reaction temperature is 0-50° C., and the reaction time is 1-8 h.

[0016] More preferably, the reaction temperature is 20-25° C., and the reaction time is 3-4 h.

[0017] Preferably, the molar ratio of the acyl chloride or sulfonyl chloride compound to 5-isosorbide mononitrate is (1-5):1.

[0018] More preferably, the molar ratio of the acyl chloride or sulfonyl chloride compound to 5-isosorbide mononitrate is (3-4):1.

[0019] Preferably, dichloromethane (DCM), triethylamine and 4-dimethylaminopyridine are added to 5-isosorbide mononitrate, and finally an acyl chloride or sulfonyl chloride compound is added to carry out the reaction.

[0020] More preferably, the molar ratio of triethylamine, 4-dimethylaminopyridine and 5-isosorbide mononitrate is (1-5): (0.1-1):1.

[0021] More preferably, the molar ratio of triethylamine, 4-dimethylaminopyridine and 5-isosorbide mononitrate is (2-4): (0.5-0.8):1.

[0022] Preferably, after the reaction, the mixture is filtered and the filtrate is dried by spin drying and passed through a column to obtain a white solid product, which is the mononitroisosorbide derivative.

[0023] An application of the mononitroisosorbide derivative is to use the mononitroisosorbide derivative in preparing antitumor drugs.

[0024] Compared with the prior art, the present invention has the following beneficial effects:

[0025] 1. This invention, based on the design concept of covalent inhibitors, has designed and synthesized a number of novel structural compounds, which is beneficial to the research and development of anti-tumor drugs;

[0026] 2. ZM533 of the present invention has good anti-tumor activity against A549 cell lines and is expected to be used in the preparation of anti-lung cancer drugs;

[0027] 3. The preparation method of the present invention is simple, the raw materials are readily available, and it is easy to scale up production. BRIEF DESCRIPTION OF THE DRAWINGS

[0028] Figure 1 This is the hydrogen nuclear magnetic resonance spectrum of the product of Example 1 of the present invention;

[0029] Figure 2 This is the hydrogen nuclear magnetic resonance spectrum of the product of Example 2 of the present invention;

[0030] Figure 3 This is the hydrogen nuclear magnetic resonance spectrum of the product of Example 3 of the present invention;

[0031] Figure 4 This is the hydrogen nuclear magnetic resonance spectrum of the product of Example 4 of the present invention;

[0032] Figure 5 This is the hydrogen nuclear magnetic resonance spectrum of the product of Example 5 of the present invention. DETAILED DESCRIPTION

[0033] The present invention is described in detail below with reference to the accompanying drawings and specific embodiments. This embodiment is implemented based on the technical solution of the present invention, and provides a detailed implementation method and specific operation process, but the protection scope of the present invention is not limited to the following embodiments.

[0034] The reagents used in the experiment were purchased from chemical reagent companies such as Titan, Bid, and Leyan. The nuclear magnetic resonance used was 600MHz and 500MHz models from Bruke, Germany, with TMS as the internal standard and DMSO- d6 , CDCl3 or D2O are used as solvents. The coupling constant (J) and chemical shift (δ) are expressed in Hz and ppm, respectively.

[0035] The raw materials and specifications used in the following examples are shown in Table 1:

[0036] Table 1 Experimental reagents, specifications and production units

[0037] name Specification Production Unit Acryloyl chloride AR Shanghai Titan Pharmaceutical Technology Co., Ltd. Sulfamyl chloride AR Shanghai Titan Pharmaceutical Technology Co., Ltd. Methacryloyl chloride AR Shanghai Titan Pharmaceutical Technology Co., Ltd. 5-Isosorbide mononitrate AR Shanghai Titan Pharmaceutical Technology Co., Ltd. Triethylamine AR McLean Biochemical Technology Co., Ltd. dichloromethane AR Greagent 4-Dimethylaminopyridine AR Shanghai Titan Pharmaceutical Technology Co., Ltd. Vinylsulfonyl chloride AR Shanghai Titan Pharmaceutical Technology Co., Ltd. p-Toluenesulfonyl chloride AR Shanghai Titan Pharmaceutical Technology Co., Ltd.

[0038] Example 1

[0039] Synthesis of ZM532:

[0040]

[0041] Place 100 mg of isosorbide 5-mononitrate in a 25 mL single-necked bottle, dissolve in 5 mL of dry DCM, and stir. Then, add 216 μL of triethylamine and 50 mg of DMAP. Finally, add 198 mg of aminosulfonyl chloride and allow to react at room temperature for 3 hours. A white solid precipitates during the reaction, and TLC (PE:EA = 1:1) indicates complete reaction. Filter the filtrate, spin dry, and pass it through a column to yield 42 mg of a white solid product (30% yield).

[0042] 1 H NMR (300MHz, DMSO) δ7.77(s,2H),5.52(t,J=4.4Hz,1H),5.00(t,J=5.3Hz,1H),4.89(d,J=2.7Hz,1H),4.58(d,J=4.9 Hz,1H),4.08(d,J=10.9Hz,1H),4.00(d,J=11.4Hz,1H),3.88(dd,J=11.5,5.2Hz,1H),3.78(dd,J=10.9,3.0Hz,1H).

[0043] Example 2

[0044] Synthesis of ZM533:

[0045]

[0046] Place 100mg of isosorbide 5-mononitrate in a 25mL single-necked flask, dissolve in 5mL of dry DCM, and stir. Add 216µL of triethylamine and 50mg of DMAP, and finally, 217mg of vinylsulfonyl chloride. Allow to react at room temperature for 3 hours. A white solid precipitates during the reaction, and TLC (PE:EA = 1:1) indicates complete reaction. Filter, and the filtrate is dried and passed through a column to yield 65mg of a pale yellow solid (45% yield).

[0047] 1 H NMR(600MHz, CDCl3) δ6.58(dd,J=16.6,9.8Hz,1H),6.48(dd,J=16.6,0.4Hz,1H),6.22–6.17(m,1H),5.37(td,J=5.5,2.7Hz,1H),5.03(t,J=5.2 Hz,1H),5.00(d,J=3.1Hz,1H),4.66(d,J=4.9Hz,1H),4.19(d,J=11.3Hz,1H),4.01(ddd,J=21.5,11.3,2.9Hz,2H),3.92(dd,J=11.3,5.6Hz,1H).

[0048] Example 3

[0049] Synthesis of ZM534:

[0050]

[0051] Place 100mg of isosorbide 5-mononitrate in a 25mL single-necked flask, dissolve in 5mL of dry DCM, and stir. Then, add 216µL of triethylamine and 50mg of DMAP. Finally, add 327mg of p-toluenesulfonyl chloride and allow to react at room temperature for 3 hours. A white solid precipitates during the reaction, and TLC (PE:EA = 1:1) indicates complete reaction. Filter the filtrate, spin dry, and pass it through a column to obtain 74mg of a white solid, with a 41% yield.

[0052] 1 H NMR (600MHz, CDCl3) δ7.82–7.77(m,2H),7.37(d,J=8.0Hz,2H),5.35–5.31(m,1H),5.00–4.92 (m,2H),4.56(d,J=4.9Hz,1H),3.98(dd,J=11.3,2.7Hz,2H),3.89–3.85(m,2H),2.47(s,3H).

[0053] Example 4

[0054] Synthesis of ZM535:

[0055]

[0056] Place 100mg of isosorbide 5-mononitrate in a 25mL single-necked flask, dissolve in 5mL of dry DCM, and stir. Then, add 216µL of triethylamine and 50mg of DMAP. Finally, add 155mg of acryloyl chloride and allow to react at room temperature for 3 hours. A white solid precipitates during the reaction, and TLC (PE:EA = 1:1) indicates complete reaction. Filter the filtrate, spin dry, and pass it through a column to obtain 46mg of a white solid, a 36% yield.

[0057] 1 H NMR (600MHz, CDCl3) δ6.45(dd,J=17.3,1.2Hz,1H),6.12(dd,J=17.3,10.5Hz,1H),5.90(dd,J=10.5,1.3Hz,1H),5.37(td,J=5.6, 2.9Hz,1H),5.31(d,J=3.1Hz,1H),5.01(t,J=5.2Hz,1H),4.53(d,J=4.9Hz,1H),4.09–4.02(m,3H),3.92(dd,J=11.3,5.6Hz,1H).

[0058] Example 5

[0059] Synthesis of ZM536:

[0060]

[0061] Place 100mg of isosorbide 5-mononitrate in a 25mL single-necked flask, dissolve in 5mL of dry DCM, and stir. Then, add 216µL of triethylamine and 50mg of DMAP. Finally, add 179mg of methacryloyl chloride and allow to react at room temperature for 3 hours. A white solid precipitates during the reaction, and TLC (PE:EA = 1:1) indicates complete reaction. Filter the filtrate, spin dry, and pass it through a column to obtain 67mg of a white solid, with a yield of 67%.

[0062] 1 H NMR (600MHz, CDCl3) δ6.13–6.11(m,1H),5.63–5.61(m,1H),5.36(td,J=5.6,2.9Hz,1H),5.29(d,J=2.9Hz,1H),5.01( t,J=5.2Hz,1H),4.54(d,J=4.9Hz,1H),4.09–4.03(m,3H),3.92(dd,J=11.3,5.6Hz,1H),1.94(dd,J=1.5,1.0Hz,3H).

[0063] (1) Evaluation of antitumor activity in vitro

[0064] A549 (human non-small cell lung cancer cell line) was selected for in vitro antitumor activity test using the CCK-8 method, with doxorubicin (DOX) as the positive control drug.

[0065] 1.1 Cell experiment reagents and instruments

[0066] The reagents and specifications used are shown in Table 2:

[0067] Table 2 Experimental reagents, specifications and production units

[0068] Reagents Specification Production Unit DMEM / F-12 medium 500mL Hyclone Fetal bovine serum 500mL GIbco PBS buffer 500mL Hyclone pancreatic enzymes 500mL Hyclone DMSO 500μL Hyclone Dual antibody 20mL GIbco CCK-8 reagent 5mL Promega

[0069] The instruments used are shown in Table 3:

[0070] Table 3 Experimental instruments, models and production units

[0071]

[0072] 1.2 In vitro cell experiments

[0073] In vitro cell experiments specifically include the following steps:

[0074] (1) Sample configuration

[0075] Preparation of test compound sample solution: Dissolve in DMSO (Merck), add culture medium containing FBS to make a 100 μL solution or uniform suspension, then dilute with culture medium containing 0.1% DMSO to the corresponding concentration. Prepare the control solution of the positive drug under the same conditions.

[0076] (2) Test method

[0077] A concentration of 7 × 10 3 Add 100 μL of a cell suspension containing 100 cells / mL to the plate and place in a 37°C, 5% CO2 incubator. After 24 hours, discard the culture medium from the 96-well plate and add 200 μL / well of the test compound sample and control solution, respectively, in triplicate. Place the 96-well plate in a 37°C, 5% CO2 incubator for 72 hours.

[0078] (3) Detection method

[0079] The CCK-8 assay was used. A mixture of 10% CCK-8 reagent was added to basal culture medium. The remaining culture medium in the 96-well plate was discarded, and the mixture was added at 100 μL / well. The 96-well plate was incubated in a 37°C, 5% CO2 incubator for 2-4 hours. Fluorescence OD values were measured using a microplate reader (Biotech) at wavelengths of 530 / 40 and 590 / 35 nm.

[0080] Cell growth inhibition rate IC% = (OD value of blank control well - OD value of drug-treated well) / OD value of blank control well × 100%. Based on the IC% values of each concentration, linear regression was performed using GraphPad software to calculate the drug concentration that inhibited cell growth by 50%, i.e., IC 50 .

[0081] The results are shown in Table 4:

[0082] Table 4 In vitro antitumor activity

[0083]

[0084] It can be seen from the data in the table that ZM533 has good anti-tumor activity against A549 cell lines and is expected to be used in the preparation of anti-lung cancer drugs.

[0085] The above description of the embodiments is intended to facilitate understanding and use of the invention by those skilled in the art. It will be apparent that those skilled in the art can readily make various modifications to these embodiments and apply the general principles described herein to other embodiments without requiring inventive effort. Therefore, the present invention is not limited to the above-described embodiments. Improvements and modifications made by those skilled in the art based on the disclosure of the present invention, without departing from the scope of the present invention, should be within the scope of protection of the present invention.

Claims

1. A mononitroisosorbide derivative, characterized in that Has the following structure: ； Wherein, R is the following structure: 。 2. The method for preparing a mononitroisosorbide derivative according to claim 1, wherein Under the action of triethylamine and catalyst 4-dimethylaminopyridine, 5-isosorbide mononitrate is reacted with a sulfonyl chloride compound to prepare the mononitroisosorbide derivative; The sulfonyl chloride compound is vinylsulfonyl chloride.

3. The method for preparing a mononitroisosorbide derivative according to claim 2, wherein: The reaction temperature is 0~50℃ and the reaction time is 1~8h.

4. The method for preparing a mononitroisosorbide derivative according to claim 2, wherein: The molar ratio of the 5-isosorbide mononitrate to the sulfonyl chloride compound is 1:(1-5).

5. The method for preparing a mononitroisosorbide derivative according to claim 2, wherein: Dichloromethane, triethylamine and 4-dimethylaminopyridine are added to 5-isosorbide mononitrate, and finally a sulfonyl chloride compound is added to carry out a reaction.

6. The method for preparing a mononitroisosorbide derivative according to claim 5, wherein: The molar ratio of triethylamine, 4-dimethylaminopyridine and 5-isosorbide mononitrate is (1-5): (0.1-1):

1.

7. The method for preparing a mononitroisosorbide derivative according to claim 2, wherein: After the reaction, the mixture was filtered, and the filtrate was dried by rotary column to obtain a white solid product, which was the mononitroisosorbide derivative.

8. A use of the mononitroisosorbide derivative according to claim 1, characterized in that: The mononitroisosorbide derivative is used for preparing an anti-non-small cell lung cancer drug.

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