Urea compound containing nitrobenzene, preparation method therefor, and use thereof

Abstract

Disclosed are a urea compound containing nitrobenzene, a preparation method therefor, and use thereof. The preparation method comprises: dissolving bis(trichloromethyl)carbonate in anhydrous tetrahydrofuran, adding a solution of 4-nitroaniline in tetrahydrofuran, stirring uniformly after dropwise addition is completed, then adding triethylamine, stirring uniformly, evaporating the solvent to dryness, adding a solution of 4-aminobenzotrifluoride in anhydrous tetrahydrofuran, and reacting at 45 °C for 3 h to obtain the urea compound containing nitrobenzene. The urea compound containing nitrobenzene can effectively alleviate the swelling and plasma extravasation of the soles of mice, and skin vasodilation and mast cell degranulation of the soles of mice, which are caused by C48 / 80, thereby inhibiting the occurrence of anaphylactoid reactions. Therefore, using the urea compound containing nitrobenzene as an anti-drug allergy formulation provides more possible treatment regimens for the clinical treatment of drug allergies.

Description

A ure compound containing nitrobenzene and a preparation method and application thereof TECHNICAL FIELD

[0001] The present application belongs to the technical field of biological medicine, and particularly relates to a ure compound containing nitrobenzene and a preparation method and application thereof. BACKGROUND

[0002] Drug allergy reaction refers to a special systemic reaction caused by drugs entering the body through oral administration, injection, enema or other routes, which is often manifested as skin redness, itching, skin rash, difficulty breathing, and even shock or death in severe cases. Drug allergy reaction belongs to non-immune allergic reaction, which is caused by direct activation of mast cells by drugs without IgE and other immunoglobulin mediation. Studies have shown that MRGPRX2 is a key receptor for mediating drug allergy reaction. When drugs act on MRGPRX2 on mast cells, the mast cells are activated to release allergic mediators and inflammatory factors, etc., which can cause skin swelling, plasma extravasation, skin vasodilation, etc. Antagonizing the activation of MRGPRX2 by drugs can significantly inhibit and treat drug allergy reaction. However, there is no marketed drug targeting this target in the clinic at present. Therefore, the development of compounds targeting MRGPRX2 can provide a new way for the treatment of drug allergy reaction.

[0003] MRGPRB2 is a homologous receptor of MRGPRX2 distributed on mouse mast cells. After drugs act on MRGPRB2 receptor, the mouse can produce allergic reactions, including skin swelling, plasma extravasation, skin vasodilation, etc. Therefore, mouse paw swelling and plasma extravasation experiments and mouse paw skin section staining experiments are often used in the laboratory to evaluate drug allergy reaction. Therefore, it is of great significance to prepare compounds that can antagonize mouse paw swelling and plasma extravasation, relieve mouse paw skin vasodilation and mast cell degranulation for the treatment of drug allergy reaction. TECHNICAL PROBLEM

[0004] However, the existing compounds have the problem of insufficient anti-allergic activity.

[0005] In order to overcome the problem of insufficient anti-allergic activity of the existing compounds in the prior art, the present application aims to provide a ure compound containing nitrobenzene and a preparation method and application thereof. TECHNICAL SOLUTION

[0006] In order to achieve the above-mentioned purpose, the present application adopts the following technical solutions:

[0007] A ure compound containing nitrobenzene, the structure of the ure compound containing nitrobenzene is as follows:

[0008] .

[0009] A preparation method of a nitrobenzene-containing urea compound, comprising the following steps:

[0010] Dissolve bis (trichloromethyl) carbonate in anhydrous tetrahydrofuran, add a tetrahydrofuran solution of 4-nitroaniline, dropwise add and stir until uniform, then add triethylamine, stir until uniform, add an anhydrous tetrahydrofuran solution of 4-amino-trifluorotoluene after evaporating the solvent, and react at 45 DEG C for 3 h to obtain the nitrobenzene-containing urea compound.

[0011] Further, the molar ratio of bis (trichloromethyl) carbonate to 4-nitroaniline is 2:5.

[0012] Further, the molar ratio of bis (trichloromethyl) carbonate to triethylamine is 1:3 mmol.

[0013] Further, the molar ratio of bis (trichloromethyl) carbonate to 4-amino-trifluorotoluene is 2:5.

[0014] A nitrobenzene-containing urea compound in the preparation of a drug for antagonizing C48 / 80-induced mouse paw swelling and plasma extravasation.

[0015] A nitrobenzene-containing urea compound in the preparation of a drug for antagonizing C48 / 80-induced mouse paw skin vasodilation and mast cell degranulation.

[0016] An anti-drug allergic reaction drug made by adding a pharmaceutically acceptable pharmaceutical excipient to a nitrobenzene-containing urea compound.

[0017] Further, the anti-allergic drug is a clinically acceptable pharmaceutical preparation.

[0018] Further, the pharmaceutical preparation is a tablet, a capsule, a granule or an injection. Beneficial effects

[0019] The present application discloses for the first time that a nitrobenzene-containing urea compound can effectively relieve C48 / 80-induced mouse paw swelling and plasma extravasation, mouse paw skin vasodilation and mast cell degranulation, thereby inhibiting the occurrence of allergic reactions, so that the nitrobenzene-containing urea compound is used as an anti-drug allergic reaction preparation, providing more possible treatment options for clinical drug allergic reaction treatment. BRIEF DESCRIPTION OF DRAWINGS

[0020] Figure 1 is the effect of a nitrobenzene-containing urea compound on mouse paw swelling and plasma extravasation, wherein (a) is a representative picture of paw swelling and Evans blue extravasation; (b) is the paw swelling rate; and (c) is the Evans blue extravasation rate.

[0021] Figure 2 is a mouse local skin H&E staining and toluidine blue staining, wherein (a) is a mouse local skin H&E staining; (b) is a mouse local skin toluidine blue staining. Embodiments of the present application

[0022] In order to enable those skilled in the art to better understand the present application, the technical solutions in the embodiments of the present application will be described clearly and completely below with reference to the drawings in the embodiments of the present application. Obviously, the described embodiments are only a part of the embodiments of the present application, rather than all the embodiments. Based on the embodiments in the present application, all other embodiments obtained by those skilled in the art without creative efforts shall fall within the scope of the present application.

[0023] It should be noted that the terms "first", "second" and the like in the specification and claims of the present application and the above-described drawings are used to distinguish similar objects, and do not necessarily indicate a specific order or a chronological sequence. It should be understood that the data thus used can be interchanged under appropriate circumstances, so that the embodiments of the present application described herein can be implemented in an order other than that illustrated or described herein. In addition, the terms "include" and "have" and any variations thereof are intended to cover non-exclusive inclusion, for example, a process, method, system, product or device that includes a series of steps or units need not be limited to only those steps or units clearly listed, but can include other steps or units not clearly listed or inherent to these processes, methods, products or devices.

[0024] The urea compound has a proton acceptor and a proton donor, can form a hydrogen bond interaction with a drug target, can act as an anion acceptor and an anion transporter, and can form a strong hydrogen bond with different anions; the strong electron-withdrawing effect of the nitro group can improve the binding force of the drug with the negative charge region of the active pocket of the MRGPRX2 target, so that the compound molecule has both urea and nitro structures, which is expected to improve the anti-drug allergy activity and improve the shortcomings of insufficient activity of existing compounds.

[0025] The urea compound is widely used as a herbicide and a plant growth regulator in agriculture, and is often used as an important intermediate of anticancer drugs, antibacterial drugs and the like in the medical field, but there are few reports on its anti-allergy related research. The compound B-1 designed and synthesized by introducing a nitro electron-withdrawing group into urea as a mother nucleus has good anti-allergy activity.

[0026] The present application will be described in further detail below with reference to the accompanying drawings.

[0027] The urea compound containing nitrobenzene in the present application is 1-(4-nitrophenyl)-3-(4-(trifluoromethyl)phenyl)urea diarylurea compound B-1, and the structural formula is as follows:

[0028]

[0029] The present application uses in vivo pharmacological experiment, adopts mouse model, and investigates the inhibition effect of the urea compound B-1 containing nitrobenzene on the mouse paw swelling and plasma extravasation, the vasodilation of the mouse paw skin and the mast cell degranulation caused by C48 / 80.

[0030] The research results show that the urea compound B-1 containing nitrobenzene can effectively relieve the allergic symptoms of the mouse paw swelling and plasma extravasation caused by C48 / 80, can weaken the vasodilation of the mouse paw skin and inhibit the mast cell degranulation, and it is proved that the urea compound B-1 containing nitrobenzene has potential anti-drug allergic reaction activity and has the prospect of being made into an anti-allergic reaction drug.

[0031] The anti-drug allergic reaction drug is a clinically acceptable pharmaceutical preparation.

[0032] Further, the pharmaceutical preparation is a tablet, a capsule, a granule or an injection.

[0033] Example 1

[0034] The synthesis steps and structure analysis nuclear magnetic data of the urea compound B-1 containing nitrobenzene are as follows:

[0035] The urea compound B-1 containing nitrobenzene: at room temperature, bis(trichloromethyl) carbonate (BTC) (1.19 g, 4 mmol) is dissolved in 10 mL of anhydrous tetrahydrofuran, and a tetrahydrofuran solution of 4-nitroaniline (1.38 g, 10 mmol) is slowly added through a constant pressure dropping funnel, and after the addition is completed, it is stirred for 15 min, and then triethylamine (1.21 g, 12 mmol) is continuously added dropwise, and after the addition is completed, it is stirred for 15 min. The solvent is evaporated, a 4-amino-trifluorotoluene (1.61 g, 10 mmol) solution dissolved in 20 mL of anhydrous tetrahydrofuran is added dropwise to the reaction solution, and after the addition is completed, it is heated to 45 ℃ and reacted for 3 h. The reaction liquid is extracted with dichloromethane and water three times, dried with anhydrous Na2SO4, rotary evaporated, and purified by a chromatographic column to obtain the compound B-1 as a light yellow solid powder, with a yield of 2.42 g and a yield of 74.41%. 1 H NMR (400 MHz, DMSO) δ 9.58 (s, 1H), 9.36 (s, 1H), 8.24 – 8.17 (m, 2H), 7.75 – 7.63 (m, 6H); EI-MS (m / z): 324.2 [M] - .

[0036] Example 2

[0037] 1. Experimental materials

[0038] Instruments: Vernier caliper purchased from Mahr, Germany, 10 μL micro-sampler purchased from Shanghai Gaoge Trading Co., Ltd., FL-4019 high-speed refrigerated centrifuge purchased from Anhui Fangling Medical Equipment Co., Ltd., SB-5200DT ultrasonic cleaner purchased from Jining Keyuan Ultrasonic Equipment Co., Ltd., 800TS microplate reader purchased from BioTek Instruments, Inc., USA, and electric heating air drying oven purchased from Tianjin Test Instrument Co., Ltd.

[0039] Experimental animals: 18-22 g, healthy adult male Kunming mice, purchased from Xi'an Jiaotong Medical Department Experimental Animal Center.

[0040] Main reagents:

[0041] 1) 6% chloral hydrate solution

[0042] An appropriate amount of chloral hydrate powder was dissolved in physiological saline to obtain a 6% chloral hydrate solution.

[0043] 2) 0.4% Evans blue solution

[0044] An appropriate amount of Evans blue powder was dissolved in physiological saline to obtain a 0.4% Evans blue solution.

[0045] 3) C48 / 80 solution

[0046] An appropriate amount of positive control drug C48 / 80 powder was dissolved in dimethyl sulfoxide to obtain a final concentration of 30 mg / mL of C48 / 80 powder, and then diluted with physiological saline to 60 μg / mL. The solution was prepared before each use.

[0047] 4) 70% acetone-physiological saline solution

[0048] Acetone and physiological saline were mixed in a volume ratio of 7:3 (V / V) before use to obtain a 70% acetone-physiological saline solution.

[0049] 5) Sample solution

[0050] An appropriate amount of sample was dissolved in dimethyl sulfoxide (DMSO), and then anhydrous ethanol, polyethylene glycol 400 (PEG400), and triple distilled water were added in sequence to dilute to the required concentration. The volume ratio of the solvent composition DMSO-C2H5OH-PEG400-H2O was 1:9:50:40.

[0051] 2. Experimental method

[0052] 18-22 g adult mice were randomly divided into administration groups (1, 5, 10 mg / kg) and a positive group (C48 / 80 group), 5 mice in each group. Intragastric administration was performed, 200 μL of mixed solvent was intragastrically administered to the positive control group, and 200 μL of compound B-1 with concentrations of 0.1, 0.5, and 1 mg / mL was intragastrically administered to the administration groups, respectively. After 2 h, 200 μL of 6% chloral hydrate solution was intraperitoneally injected into the mice, and the mice were usually anesthetized after 15 min. 200 μL of 0.4% Evans blue solution was injected into the tail vein of the mice, and the thickness of the left and right footpads of the mice (La, Ra) was measured. After 15 min, 5 μL of C48 / 80 (with a concentration of 60 μg / mL) and normal saline were injected into the left and right footpads of the mice, respectively. After 15 min, the mice were killed by decapitation, and the thickness of the left and right footpads (Lb, Rb) was measured again. The left and right footpads of the hind limbs were photographed and recorded. The toes were cut along the elbow joint, placed in an EP tube, labeled, dried, and weighed (W). 400 μL of 70% acetone-saline was added to the EP tube, the footpad tissue was cut and ultrasonicated, centrifuged, and the supernatant was aspirated. The OD value of each well was measured at 630 nm.

[0053] Data processing:

[0054] A: Footpad swelling rate = (Lb-La) / La (left); (Rb-Ra) / Ra (right);

[0055] B: Unit Evans blue exudation = absorbance value (OD) / toe weight (W).

[0056] 3. Experimental results

[0057] The data are expressed as mean ± SEM, and the graphs are drawn using Graphpad Prism 9.0 software.

[0058] The experimental results are shown in FIG. 1. Compound 48 / 80 (C48 / 80) is a substance commonly used in biochemical research to promote mast cell degranulation. C48 / 80 is the model group, representing the mouse footpad swelling and plasma extravasation caused by the sensitizing substance C48 / 80. Different concentrations represent different experimental groups. Compound B-1 antagonizes mouse footpad swelling and tissue extravasation in the presence of C48 / 80. The experimental results show that compound B-1 can antagonize mouse footpad swelling and plasma extravasation caused by C48 / 80, and this antagonistic effect is positively correlated with the dosage.

[0059] Example 3

[0060] 1. Experimental materials

[0061] Instruments: 10 μL microsyringe purchased from Shanghai Gaoge Industry and Trade Co., Ltd., and ECLIPSE Ci-L upright imaging microscope purchased from Nikon Corporation, Japan.

[0062] Experimental animals: 18-22 g, healthy adult male Kunming mice, purchased from the Experimental Animal Center of Xi'an Jiaotong Medical College.

[0063] Main reagents:

[0064] 1) 6% chloral hydrate solution

[0065] An appropriate amount of chloral hydrate powder was dissolved in physiological saline to obtain a 6% chloral hydrate solution.

[0066] 2) C48 / 80 solution

[0067] An appropriate amount of C48 / 80 powder was dissolved in dimethyl sulfoxide to a final concentration of 30 mg / mL, and then diluted with physiological saline to 60 μg / mL. It was prepared and used immediately before each use.

[0068] 3) Sample solution

[0069] An appropriate amount of sample was dissolved in dimethyl sulfoxide (DMSO), and then anhydrous ethanol, polyethylene glycol 400 (PEG400), and triple distilled water were added in sequence to dilute it to the desired concentration. The volume ratio of the solvent composition DMSO-C2H5OH-PEG400-H2O was 1:9:50:40.

[0070] 2. Experimental method

[0071] 18-22 g adult mice were randomly divided into a drug administration group (10 mg / kg), a blank control group (normal saline group), and a positive control group (C48 / 80 group), with 5 mice in each group. Intragastric administration was performed, and the positive group was given 200 μL of mixed solvent, and the drug administration group was given 200 μL of compound B-1 at a concentration of 1 mg / mL. After 2 h, 200 μL of 6% chloral hydrate solution was injected intraperitoneally to anesthetize the mice, and the blank group and the drug administration group were injected with 5 μL of normal saline and 60 μg / mL of C48 / 80 into the left footpads of the mice, respectively. After 15 min, the mice were sacrificed, and the skin tissue of the footpads was cut off and soaked in 4% paraformaldehyde for fixation. After fixation, the skin tissue was embedded, sectioned, deparaffinized, antigen repaired and blocked; H&E staining and toluidine blue staining were performed, respectively, and the slides were sealed and photographed.

[0072] 3. Experimental results

[0073] The experimental results are shown in Figure 2, wherein C48 / 80 is a model group, representing the vasodilation of mouse paw skin and mast cell degranulation caused by the sensitizing substance C48 / 80; compound B-1 antagonizes the vasodilation of mouse paw skin and mast cell degranulation in the presence of C48 / 80. The experimental results show that compound B-1 can significantly antagonize the vasodilation of mouse paw skin and mast cell degranulation caused by C48 / 80.

[0074] The nitrobenzene-containing urea compound of the present application and its application in the preparation of an anti-allergic drug can effectively antagonize the mouse paw swelling and plasma extravasation caused by C48 / 80, the vasodilation of mouse paw skin and mast cell degranulation, reduce the occurrence of allergic symptoms, and thus inhibit the occurrence of drug allergic reactions. Therefore, the nitrobenzene-containing urea compound is used as an anti-drug allergic reaction preparation, providing more possible treatment options for clinical drug allergic reaction treatment.

[0075] The nitrobenzene-containing urea compound B-1 of the present application can effectively alleviate the mouse paw swelling and plasma extravasation caused by C48 / 80, the vasodilation of mouse paw skin and mast cell degranulation, and thus inhibit the occurrence of allergic reactions. Therefore, the nitrobenzene-containing urea compound B-1 is used as an anti-drug allergic reaction preparation, providing more possible treatment options for clinical drug allergic reaction treatment.

[0076] The above content only illustrates the technical idea of the present application and cannot limit the protection scope of the present application. Any modification made according to the technical idea of the present application on the basis of the technical solution falls within the protection scope of the claims of the present application.

Claims

1. A urea compound containing nitrobenzene, characterized in that, The structural formula of the nitrobenzene-containing urea compound is as follows: 。 2. A method for preparing a urea compound containing nitrobenzene, characterized in that, Includes the following steps: Bis(trichloromethyl) carbonate was dissolved in anhydrous tetrahydrofuran, and a tetrahydrofuran solution of 4-nitroaniline was added. After the addition was complete, the mixture was stirred until homogeneous. Then, triethylamine was added and stirred until homogeneous. After the solvent was evaporated, an anhydrous tetrahydrofuran solution of 4-aminotrifluorotoluene was added. The mixture was reacted at 45 °C for 3 h to obtain urea compounds containing nitrobenzene.

3. The method for preparing nitrobenzene-containing urea compounds according to claim 2, characterized in that, The molar ratio of bis(trichloromethyl) carbonate to 4-nitroaniline is 2:

5.

4. The method for preparing nitrobenzene-containing urea compounds according to claim 2, characterized in that, The molar ratio of bis(trichloromethyl) carbonate to triethylamine is 1:3 mmol.

5. The method for preparing nitrobenzene-containing urea compounds according to claim 2, characterized in that, The molar ratio of bis(trichloromethyl) carbonate to 4-aminotrifluorotoluene is 2:

5.

6. The use of a nitrobenzene-containing urea compound as described in claim 1 in the preparation of a drug for antagonizing C48 / 80-induced paw edema and plasma extravasation in mice.

7. The use of a nitrobenzene-containing urea compound as described in claim 1 in the preparation of a medicament for antagonizing C48 / 80-induced vasodilation and mast cell degranulation in mouse paw skin.

8. A drug for treating drug-induced allergic reactions, characterized in that, It is made from the nitrobenzene-containing urea compound of claim 1 with the addition of pharmaceutically acceptable excipients.

9. The anti-drug allergy medication according to claim 8, characterized in that, Anti-allergy medications are clinically acceptable pharmaceutical preparations.

10. The anti-drug allergy medication according to claim 8, characterized in that, Pharmaceutical preparations are tablets, capsules, granules, or injections.

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