Nitrobenzene-containing urea compound as well as preparation method and application thereof
By designing urea compounds containing nitrobenzene, the problem of insufficient anti-allergic activity of existing compounds is solved, effective antagonism of drug allergic reactions is achieved, and more effective treatment plans are provided.
Patent Information
- Application Number
- CN202410830270.9
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-06-25
- Publication Date
- 2024-12-17
- Estimated Expiration
- Not applicable · inactive patent
AI Technical Summary
Existing compounds have insufficient anti-allergic activity when antagonizing drug allergic reactions and cannot effectively inhibit drug allergic reactions.
A urea compound containing nitrobenzene was designed to prepare the compound by specific synthetic methods and applied to antagonize the swelling of the soles of mice and plasma extravasation, skin vasodilation and mast cell degranulation to improve the MRGPRX2 receptor Body binding force to enhance antiallergic activity.
This compound can effectively alleviate the swelling of the soles of mice caused by C48/80, plasma extravasation and skin vasodilation, inhibit mast cell degranulation, thereby significantly inhibiting the occurrence of drug allergic reactions, and providing a more effective treatment plan.
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Abstract
Description
Technical Field
[0001] The invention belongs to the technical field of biomedicine, and specifically relates to a urea compound containing nitrobenzene and a preparation method and application thereof. Background Art
[0002] Drug allergic reaction refers to a special systemic reaction caused by drugs entering the body through oral, injection, enema or other routes, which is often manifested as skin flushing, itching, rash, and difficulty breathing. In severe cases, shock or death may occur. Drug allergic reaction is a non-immune allergic reaction. The drug directly activates mast cells and stimulates the body to produce allergic symptoms without the mediation of immunoglobulins such as IgE. Studies have shown that MRGPRX2 is a key receptor that mediates drug allergic reactions. Drugs act on MRGPRX2 on mast cells, thereby activating mast cells and releasing allergic mediators and inflammatory factors, causing symptoms such as skin swelling, plasma extravasation, and skin vasodilation. Antagonizing the activation of MRGPRX2 by drugs can significantly inhibit and treat drug allergic reactions. However, there are currently no marketed drugs targeting this target in clinical practice, so the research and development of compounds targeting MRGPRX2 can provide a new way to treat drug allergic reactions.
[0003] MRGPRB2 is a homologous receptor of MRGPRX2 distributed on mouse mast cells. Drugs acting on MRGPRB2 receptors can induce allergic reactions in mice, including skin swelling, plasma extravasation, and skin vasodilation. Therefore, the laboratory often uses mouse paw swelling and plasma extravasation experiments and mouse paw skin section staining experiments to evaluate drug allergic reactions. Therefore, the preparation of compounds that can antagonize mouse paw swelling and plasma extravasation, relieve mouse paw skin vasodilation and mast cell degranulation is of great significance for the treatment of drug-like allergic reactions.
[0004] However, existing compounds have the problem of insufficient anti-allergic activity. Summary of the invention
[0005] In order to overcome the problem that the compounds in the above-mentioned prior art have insufficient anti-allergic activity, the purpose of the present invention is to provide a urea compound containing nitrobenzene and a preparation method and application thereof.
[0006] In order to achieve the above object, the present invention adopts the following technical solutions:
[0007] A urea compound containing nitrobenzene, the structural formula of the urea compound containing nitrobenzene is as follows:
[0008]
[0009] A method for preparing a urea compound containing nitrobenzene comprises the following steps:
[0010] Dissolve bis(trichloromethyl) carbonate in anhydrous tetrahydrofuran, add a tetrahydrofuran solution of 4-nitroaniline, stir evenly after the addition is complete, then add triethylamine, stir evenly, evaporate the solvent, add anhydrous tetrahydrofuran solution of 4-aminotrifluorotoluene, react at 45°C for 3h to obtain a urea compound containing nitrobenzene.
[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-aminotrifluorotoluene is 2:5.
[0014] The invention discloses an application of a urea compound containing nitrobenzene in preparing a drug for antagonizing C48 / 80-induced swelling of the soles of mice and plasma extravasation.
[0015] The invention discloses an application of a urea compound containing nitrobenzene in the preparation of a medicine for antagonizing vasodilation of mouse paw skin and mast cell degranulation caused by C48 / 80.
[0016] The invention discloses a drug for preventing drug allergic reaction, which is prepared by adding pharmaceutically acceptable pharmaceutical excipients to a urea compound containing nitrobenzene.
[0017] Furthermore, the anti-allergic drug is a clinically acceptable pharmaceutical preparation.
[0018] Furthermore, the pharmaceutical preparation is a tablet, capsule, granule or injection.
[0019] Compared with the prior art, the present invention has the following beneficial effects:
[0020] The present invention discloses for the first time that urea compounds containing nitrobenzene can effectively alleviate the swelling and plasma extravasation of mouse paws caused by C48 / 80, vasodilation of mouse paw skin and mast cell degranulation, thereby inhibiting the occurrence of anaphylactic-like reactions. Therefore, urea compounds containing nitrobenzene are used as anti-drug allergic reaction preparations, providing more possible treatment options for the clinical treatment of drug allergic reactions. BRIEF DESCRIPTION OF THE DRAWINGS
[0021] Figure 1 The effects of urea compounds containing nitrobenzene on mouse paw swelling and plasma extravasation. (a) Representative images of paw swelling and Evans blue extravasation; (b) Paw swelling rate; (c) Evans blue extravasation rate.
[0022] Figure 2H&E staining and toluidine blue staining of mouse local skin. (a) H&E staining of mouse local skin; (b) Toluidine blue staining of mouse local skin. DETAILED DESCRIPTION
[0023] In order to enable those skilled in the art to better understand the scheme of the present invention, the technical scheme in the embodiments of the present invention will be clearly and completely described below in conjunction with the drawings in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work should fall within the scope of protection of the present invention.
[0024] It should be noted that the terms "first", "second", etc. in the specification and claims of the present invention and the above-mentioned drawings are used to distinguish similar objects, and are not necessarily used to describe a specific order or sequence. It should be understood that the data used in this way can be interchanged where appropriate, so that the embodiments of the present invention described herein can be implemented in an order other than those illustrated or described herein. In addition, the terms "including" and "having" and any variations thereof are intended to cover non-exclusive inclusions, for example, a process, method, system, product or device that includes a series of steps or units is not necessarily limited to those steps or units that are clearly listed, but may include other steps or units that are not clearly listed or inherent to these processes, methods, products or devices.
[0025] Urea compounds have both proton acceptors and proton donors, and can form hydrogen bond interactions with drug targets. They can also act as anion receptors and anion transporters, and can form strong hydrogen bonds with different anions. The strong electron-withdrawing effect of the nitro group can enhance the binding force between the drug and the negatively charged region of the active pocket of the allergic target MRGPRX2. Therefore, the compound molecule has both urea and nitro structures, which is expected to enhance the activity against drug allergic reactions and improve the shortcomings of insufficient activity of existing compounds.
[0026] Urea compounds are widely used as herbicides and plant growth regulators in agriculture; they are often used as important intermediates for anticancer drugs, antibacterial drugs, etc. in the medical field, but there are few reports on their anti-allergic effects. The present invention uses urea as the parent nucleus and introduces a nitro electron-withdrawing group to design and synthesize compound B-1, which has good anti-allergic activity.
[0027] The present invention is further described in detail below with reference to the accompanying drawings.
[0028] The urea compound containing nitrobenzene in the present invention is 1-(4-nitrophenyl)-3-(4-(trifluoromethyl)phenyl)urea diaryl urea compound B-1, and the structural formula is as follows:
[0029]
[0030] The present invention uses an in vivo pharmacological experiment and a mouse model to investigate the inhibitory effect of a urea compound B-1 containing nitrobenzene on mouse paw swelling and plasma extravasation caused by C48 / 80, mouse paw skin vasodilation and mast cell degranulation.
[0031] The research results show that the urea compound B-1 containing nitrobenzene can effectively relieve the allergic-like symptoms of mouse paw swelling and plasma extravasation caused by C48 / 80, weaken the vasodilation of mouse paw skin and inhibit mast cell degranulation, indicating that the urea compound B-1 containing nitrobenzene has potential anti-drug allergic reaction activity and has the prospect of being made into anti-allergic reaction drugs.
[0032] The anti-drug allergic reaction drug is a clinically acceptable drug preparation.
[0033] Furthermore, the pharmaceutical preparation is a tablet, capsule, granule or injection.
[0034] Example 1
[0035] The synthesis steps and structural analysis NMR data of nitrobenzene-containing urea compound B-1 are as follows:
[0036] Urea compound B-1 containing nitrobenzene: At room temperature, bis(trichloromethyl) carbonate (BTC) (1.19 g, 4 mmol) was dissolved in 10 mL of anhydrous tetrahydrofuran, and a tetrahydrofuran solution of 4-nitroaniline (1.38 g, 10 mmol) was slowly added dropwise through a constant pressure dropping funnel. After the addition was completed, the mixture was stirred for 15 min. Triethylamine (1.21 g, 12 mmol) was continued to be added to the turbid solution. After the addition was completed, the mixture was stirred for 15 min. The solvent was evaporated, and a solution of 4-aminotrifluorotoluene (1.61 g, 10 mmol) dissolved in 20 mL of anhydrous tetrahydrofuran was added dropwise to the reaction solution. After the addition was completed, the temperature was raised to 45°C and the reaction was reacted for 3 h. The reaction solution was extracted with dichloromethane and water three times, dried over anhydrous Na2SO4, and dried by spin drying. The mixture was purified by chromatography to obtain compound B-1 as a light yellow solid powder with a yield of 2.42 g and a yield of 74.41%. 1 HNMR(400MHz, DMSO)δ9.58(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] - .
[0037] Example 2
[0038] 1. Experimental Materials
[0039] Instruments: Vernier calipers were purchased from Menite, Germany; 10 μL microinjector was purchased from Shanghai Gaoge Industry and Trade Co., Ltd.; FL-4019 high-speed refrigerated centrifuge was purchased from Anhui Fangling Medical Equipment Co., Ltd.; SB-5200DT ultrasonic cleaner was purchased from Jining Keyuan Ultrasonic Equipment Co., Ltd.; 800TS microplate reader was purchased from BioTek Instrument Co., Ltd., USA; and electric blast drying oven was purchased from Tianjin Test Instrument Co., Ltd.
[0040] Experimental animals: 18-22 g, healthy adult male Kunming mice were purchased from the Experimental Animal Center of Xi'an Jiaotong Medical College.
[0041] Main reagents:
[0042] 1) 6% chloral hydrate solution
[0043] Weigh an appropriate amount of chloral hydrate powder and dissolve it in physiological saline to obtain a chloral hydrate solution with a mass concentration of 6%.
[0044] 2) 0.4% Evans blue solution
[0045] Weigh an appropriate amount of Evans blue powder and dissolve it in physiological saline to obtain an Evans blue solution with a mass concentration of 0.4%.
[0046] 3) C48 / 80 solution
[0047] Weigh an appropriate amount of positive control drug C48 / 80 powder, dissolve it in dimethyl sulfoxide to make the final concentration of C48 / 80 powder 30 mg / mL, dilute it to 60 μg / mL with normal saline, prepare it before each use, and use it immediately.
[0048] 4) 70% acetone-saline solution
[0049] Acetone and physiological saline were mixed at a volume ratio of 7:3 (V / V) before use to obtain an acetone-physiological saline solution with a volume concentration of 70%.
[0050] 5) Sample solution
[0051] Weigh an appropriate amount of sample, dissolve it in dimethyl sulfoxide (DMSO), and add anhydrous ethanol, polyethylene glycol 400 (PEG400), and triple distilled water to dilute to the required concentration. The solvent composition is DMSO-C2H5OH-PEG400-H2O with a volume ratio of 1:9:50:40.
[0052] 2. Experimental Methods
[0053] Adult mice weighing 18-22g were randomly divided into a dosing group (1, 5, 10mg / kg) and a positive group (C48 / 80 group), with 5 mice in each group. The positive control group was gavaged with 200μL of mixed solvent, and the dosing group was gavaged with 200μL of compound B-1 at a concentration of 0.1, 0.5, and 1mg / mL. After 2h, 200μL of 6% chloral hydrate solution was intraperitoneally injected into the mouse. Usually 15min later, the mouse was anesthetized, and 200μL of 0.4% Evans blue solution was injected into the tail vein to measure the thickness of the left and right soles of the mouse (La, Ra). After 15min, 5μL C48 / 80 (concentration of 60μg / mL) and saline were injected into the left and right soles of the mouse, respectively. After 15min, the mouse was killed by dislocation of the neck, and the thickness of the left and right soles (Lb, Rb) was measured again, and the left and right soles of its hind limbs were photographed and recorded. Cut the toes along the elbow joint, put them in an EP tube, mark them with numbers, dry them, and weigh them (W). Add 400 μL of 70% acetone-normal saline to the EP tube, cut the sole tissue into pieces and sonicate, centrifuge, absorb the supernatant, and measure the OD value of each well at 630 nm.
[0054] Data processing:
[0055] A: Foot swelling rate = (Lb-La) / La (left); (Rb-Ra)Ra (right);
[0056] B: Unit Evans blue leakage = absorbance value (OD) / toe weight (W).
[0057] 3. Experimental results
[0058] The data are expressed as mean ± SEM, and the graphs were drawn using Graphpad Prism 9.0 software.
[0059] The experimental results are shown in Figure 1 Among them, 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 swelling of the mouse paw and plasma extravasation caused by the sensitizer C48 / 80; different concentrations represent different experimental groups. Compound B-1 antagonizes the swelling of the mouse paw and tissue extravasation in the presence of C48 / 80. The experimental results show that compound B-1 can antagonize the swelling of the mouse paw and plasma extravasation caused by C48 / 80, and this antagonistic effect is positively correlated with the dosage.
[0060] Example 3
[0061] 1. Experimental Materials
[0062] Instruments: 10 μL microinjector was purchased from Shanghai Gaoge Industry and Trade Co., Ltd., and ECLIPSE Ci-L upright imaging microscope was purchased from Nikon Corporation of Japan.
[0063] Experimental animals: 18-22 g, healthy adult male Kunming mice were purchased from the Experimental Animal Center of Xi'an Jiaotong Medical College.
[0064] Main reagents:
[0065] 1) 6% chloral hydrate solution
[0066] Weigh an appropriate amount of chloral hydrate powder and dissolve it in physiological saline to obtain a chloral hydrate solution with a mass concentration of 6%.
[0067] 2) C48 / 80 solution
[0068] Weigh an appropriate amount of C48 / 80 powder, dissolve it in dimethyl sulfoxide to a final concentration of 30 mg / mL, dilute it to 60 μg / mL with normal saline, prepare it before each use, and use it immediately after preparation.
[0069] 3) Sample solution
[0070] Weigh an appropriate amount of sample, dissolve it in dimethyl sulfoxide (DMSO), and add anhydrous ethanol, polyethylene glycol 400 (PEG400), and triple distilled water to dilute to the required concentration. The solvent composition is DMSO-C2H5OH-PEG400-H2O with a volume ratio of 1:9:50:40.
[0071] 2. Experimental Methods
[0072] Adult mice weighing 18-22g were randomly divided into a dosing group (10mg / kg), a blank control group (normal saline group) and a positive control group (C48 / 80 group), with 5 mice in each group. The positive group was gavaged with 200μL of mixed solvent, and the dosing group was gavaged with 200μL of compound B-1 at a concentration of 1mg / mL. After 2h, 200μL of 6% chloral hydrate solution was intraperitoneally injected into the mice to anesthetize them. The blank group and the dosing group were injected with 5μL of normal saline and 60μg / mL C48 / 80 into the left sole of the mouse, respectively. After 15min, the mice were killed, the skin tissue of the sole was cut off, and it was fixed by immersion in 4% paraformaldehyde. After fixation, the skin tissue was embedded, sliced, deparaffinized, antigen repaired and blocked; H&E staining and toluidine blue staining were performed, the slides were sealed and photographed.
[0073] 3. Experimental results
[0074] The experimental results are shown in Figure 2In the experiment, C48 / 80 was the model group, representing the vasodilation and mast cell degranulation of the mouse paw skin caused by the sensitizer C48 / 80; compound B-1 antagonized the vasodilation and mast cell degranulation of the mouse paw skin in the presence of C48 / 80. The experimental results showed that compound B-1 could significantly antagonize the vasodilation and mast cell degranulation of the mouse paw skin caused by C48 / 80.
[0075] The urea compounds containing nitrobenzene of the present invention and their use in the preparation of antiallergic drugs can effectively antagonize the swelling of the soles of mice and plasma extravasation, vasodilation of the skin of the soles of mice and mast cell degranulation caused by C48 / 80, reduce the occurrence of allergic-like symptoms, and thus inhibit the occurrence of drug allergic reactions. Therefore, the urea compounds containing nitrobenzene are used as anti-drug allergic reaction preparations, providing more possible treatment options for the clinical treatment of drug allergic reactions.
[0076] The present invention discloses for the first time that a urea compound B-1 containing nitrobenzene can effectively alleviate the swelling and plasma extravasation of the soles of mice caused by C48 / 80, vasodilation of the skin blood vessels of the soles of mice and degranulation of mast cells, thereby inhibiting the occurrence of anaphylactic-like reactions. Therefore, the urea compound B-1 containing nitrobenzene is used as an anti-drug allergic reaction preparation, providing more possible treatment options for the clinical treatment of drug allergic reactions.
[0077] The above contents are only for explaining the technical idea of the present invention and cannot be used to limit the protection scope of the present invention. Any changes made on the basis of the technical solution in accordance with the technical idea proposed by the present invention shall fall within the protection scope of the claims of the present invention.
Claims
1. A urea compound containing nitrobenzene, characterized in that: The structural formula of the urea compound containing nitrobenzene is as follows:
2. A method for preparing a urea compound containing nitrobenzene, characterized in that: The following steps are involved: Dissolve bis(trichloromethyl) carbonate in anhydrous tetrahydrofuran, add a tetrahydrofuran solution of 4-nitroaniline, stir evenly after the addition is complete, then add triethylamine, stir evenly, evaporate the solvent, add anhydrous tetrahydrofuran solution of 4-aminotrifluorotoluene, react at 45°C for 3h to obtain a urea compound 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 was 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-aminobenzotrifluoride is 2:
5.
6. Use of the nitrobenzene-containing urea compound as claimed in claim 1 in the preparation of a drug for antagonizing C48 / 80-induced swelling of the paws and plasma extravasation in mice.
7. Use of the nitrobenzene-containing urea compound according to claim 1 in the preparation of a drug for antagonizing C48 / 80-induced vasodilation in the skin of the soles of mice and degranulation of mast cells.
8. A drug for preventing drug allergic reactions, characterized in that: The urea compound containing nitrobenzene according to claim 1 is prepared by adding pharmaceutically acceptable excipients.
9. The anti-drug allergy drug according to claim 8, characterized in that: Anti-allergic drugs are clinically acceptable pharmaceutical preparations.
10. The drug for preventing drug allergic reaction according to claim 8, characterized in that: The pharmaceutical preparations are tablets, capsules, granules or injections.