A water-soluble expanded biphenyl[3]arene derivative, a preparation method and application thereof

Abstract

The application provides a water-soluble expanded biphenyl[3]arene derivative and a preparation method and application thereof, and belongs to the technical field of chemical medicine manufacturing. The water-soluble (expanded) biphenyl[3]arene derivative is prepared by a nucleophilic substitution reaction of full-hydroxyl (expanded) biphenyl[3]arene as raw material and propane sulfonic acid lactone in a dioxane / water mixed solution. The (expanded) biphenyl[3]arene is suitable for capsaicin and is easy to be post-modified as a macrocyclic skeleton, the flexible alkyl side chain is introduced to prolong the cavity depth, expand the area of hydrophobic interaction and enhance the complexing capacity, and the terminal sodium sulfonate group is arranged to improve the water solubility and biocompatibility of the molecule, so that the capsaicin can be wiped and removed by the recognition and solubilization, and the stimulation of the capsaicin on cells can be inhibited by the smearing and blending, the cytotoxicity of the capsaicin on human immortalized epidermal cells is reduced, and the scratching behavior caused by the capsaicin stimulation is relieved.

Description

Technical Field

[0001] This invention relates to the field of chemical manufacturing technology, and in particular to a water-soluble extended biphenyl[3] aromatic derivative, its preparation method and application. Background Technology

[0002] Capsaicin, with the structure trans-8-methyl-N-vanillyl-6-nonenamide, is the main active ingredient in chili peppers and is widely used in various industries as a food additive, pharmaceutical molecule, antifouling coating, and insect and rodent repellent. Capsaicin is highly irritating and can cause symptoms such as eye pain, tearing, coughing, sneezing, and chest pain at low thresholds. It is also used as an irritant by activating the transient receptor potential vanillin receptor 1 (TRPV1) in the human body, triggering an influx of calcium ions, thereby producing a burning sensation and pain signals.

[0003] In daily life, emergency treatment for small amounts of capsaicin contact with the skin mainly relies on physical removal (such as wiping with oil or alcohol) and symptom relief (cold compresses, anti-inflammatory drugs). In cases of large exposure, medications can be used to block the binding of capsaicin to TRPV1, inhibit the inflammatory response, or local anesthesia can be used to alleviate the irritation. However, all of the above methods have some drawbacks. For example, capsaicin, as a fat-soluble alkaloid, easily binds to lipids in the stratum corneum of the skin to form stable complexes. Although alcohol or oil can dissolve surface capsaicin, it cannot penetrate deep into the epidermis to remove molecules embedded in the dermis. Studies have shown that untreated capsaicin can remain in the skin for more than 48 hours, continuously activating TRPV1 receptors and causing neurogenic inflammation. Drug treatment mainly focuses on the "symptoms" caused by capsaicin, but does not address the "root cause" of capsaicin molecules.

[0004] Currently, supramolecular isolation strategies hold promise for solving the aforementioned challenges. This method primarily relies on molecular recognition, using pharmacokinetic mechanisms to reduce the free concentration of the complexed compound or inhibit its interaction with target cells. The most representative isostatic agent is sugammadextrin sodium, a γ-cyclodextrin derivative that specifically recognizes the muscle relaxant rocuronium bromide, thereby reversing muscle relaxation. Furthermore, this strategy has been successfully applied to various drugs or toxic substances with severe side effects. However, traditional macrocyclic compounds have relatively fixed sizes, making precise adaptation to the guest's structure difficult, thus limiting performance modulation. Therefore, there is an urgent need to develop a macrocyclic compound adapted to capsaicin to efficiently recognize and alleviate capsaicin-induced irritation symptoms. Summary of the Invention

[0005] The purpose of this invention is to provide a water-soluble (extended) biphenyl[3] aromatic derivative, its preparation method and application. This water-soluble (extended) biphenyl[3] aromatic derivative can increase the solubility of capsaicin in water through host-guest recognition, thereby reducing the concentration of free capsaicin or inhibiting the interaction between complexed capsaicin and receptors in a pharmacokinetic manner to alleviate related irritation symptoms.

[0006] To achieve the above-mentioned objectives, the present invention provides the following technical solution:

[0007] This invention provides a water-soluble (extended)biphenyl[3] aromatic derivative, wherein the water-soluble extended biphenyl[3] aromatic derivative is an (extended)biphenyl[3] aromatic sulfonate, and the structure of the (extended)biphenyl[3] aromatic sulfonate is shown in Formula I:

[0008]

[0009] The present invention also provides a method for preparing a water-soluble (extended) biphenyl[3] aromatic derivative, comprising the following steps:

[0010] The all-hydroxy (extended) biphenyl[3] aryl hydrocarbon was subjected to a nucleophilic substitution reaction with propane sulfonate lactone in a mixture of 1,4-dioxane and water in the presence of sodium hydride. The solution after the nucleation substitution reaction was dialyzed and dried to obtain (extended) biphenyl[3] aryl sulfonate.

[0011] Preferably, the molar and volume ratio of the mixture of the fully hydroxyl extended biphenyl[3] aromatic hydrocarbon, sodium hydride, propane sulfonate lactone, 1,4-dioxane and water is 0.1-0.2 mmol: 4-6 mmol: 4-6 mmol: 8-10 mL.

[0012] Preferably, in the mixture of 1,4-dioxane and water, the volume ratio of 1,4-dioxane to water is 1-2:1-2.

[0013] Preferably, the nucleophilic substitution reaction is carried out at a temperature of 85–95°C for a time of 12–36 h.

[0014] Preferably, the dialysis method is as follows:

[0015] Place the solution in a dialysis bag, place the dialysis bag in water and stir. After dialysis, take the solution from the dialysis bag for the next step of processing.

[0016] Preferably, the molecular weight cutoff of the dialysis bag is 900-1100; and the dialysis time is 1.5-3 days.

[0017] The present invention also provides an application of the above-mentioned water-soluble (extended) biphenyl[3] aromatic derivative in alleviating the irritation caused by capsaicin.

[0018] Preferably, the application in relieving the irritation caused by capsaicin includes applying a (extended) biphenyl[3] aromatic hydrocarbon derivative solution to the exposed area of ​​capsaicin to inhibit its irritation in a pharmacokinetic manner; or, using (extended) biphenyl[3] aromatic hydrocarbon derivative as a decontaminant to remove capsaicin adhering to the skin by solubilization and wiping.

[0019] The benign solvent for the (extended) biphenyl[3] aromatic derivative is one or a mixture of water, phosphate buffer, dimethyl sulfoxide, and methanol.

[0020] The present invention also provides an application of the above-mentioned method for preparing water-soluble (extended) biphenyl[3] aromatic derivatives in the preparation of products that alleviate capsaicin irritation.

[0021] The beneficial effects of this invention compared to the prior art are as follows:

[0022] (1) This invention prepares a water-soluble (extended) biphenyl[3] aromatic derivative by reacting a fully hydroxyl (extended) biphenyl[3] aromatic hydrocarbon with propane sulfonate lactone in a dioxane / water mixture via a nucleophilic substitution reaction. The (extended) biphenyl[3] aromatic hydrocarbon serves as a macrocyclic skeleton, is compatible with capsaicin, and is easily modified. The introduction of flexible alkyl side chains aims to extend the cavity depth and expand the hydrophobic interaction area to enhance the complexing ability. The sodium sulfonate group at the end is used to improve the water solubility and biocompatibility of the molecule. Capsaicin can be removed by wiping with a solubilizing agent and inhibited by applying a blend. Experimental results show that the complexation by the water-soluble (extended) biphenyl[3] aromatic derivative can reduce the cytotoxicity of capsaicin on human immortalized epidermal cells and can also alleviate the scratching behavior caused by capsaicin stimulation in a Kunming mouse model.

[0023] (2) The water-soluble (extended) biphenyl[3] aromatic derivative of the present invention can increase the solubility of capsaicin in water through host-guest recognition, reduce the concentration of free capsaicin or inhibit the interaction between complexed capsaicin and receptor in a pharmacokinetic manner to relieve related irritation symptoms, and can act as a decontaminant to remove capsaicin attached to the epidermis by solubilizing and wiping.

[0024] (3) The water-soluble (extended) biphenyl[3] aromatic derivative of the present invention has mild and efficient reaction conditions, and is suitable for industrial production; it has good water solubility and biocompatibility, and is expected to be used in biomedicine; the method of use includes applying or wiping on the exposed part, which is convenient and easy to do, and is of great significance for the prevention and control of capsaicin-like substances. Attached Figure Description

[0025] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0026] Figure 1 This is a chemical reaction diagram of the synthesis of water-soluble (extended) biphenyl[3] aromatic derivatives in Example 1 of the present invention;

[0027] Figure 2 The 1H NMR spectrum of the water-soluble (extended) biphenyl[3] aromatic derivative in Example 1 of this invention;

[0028] Figure 3 The above is the hydrogen NMR spectrum of the water-soluble (extended) biphenyl[3] aromatic derivative and capsaicin complex in Experiment Example 1 of this invention;

[0029] Figure 4 The cytotoxicity of water-soluble (extended) biphenyl[3] aromatic derivatives to human immortalized epidermal cells in Experiment Example 2 of this invention;

[0030] Figure 5 The cytotoxicity of capsaicin and capsaicin / water-soluble (extended) biphenyl[3] aromatic derivative complex on human immortalized epidermal cells in Experiment Example 2 of this invention;

[0031] Figure 6 This is a graph showing the trend of body weight change in Kunming mice after treatment with water-soluble (extended) biphenyl[3] aromatic derivatives in Experiment Example 3 of this invention;

[0032] Figure 7 This is an organ index diagram of Kunming mice 14 days after treatment with water-soluble (extended) biphenyl[3] aromatic derivatives in Experiment Example 3 of this invention;

[0033] Figure 8 This is a statistical graph showing the scratching behavior of mice after different treatments in Experiment Example 4 of this invention. Detailed Implementation

[0034] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as a limitation of the present invention, but rather as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0035] It should be understood that the terminology used in this invention is merely for describing particular embodiments and is not intended to limit the invention. Furthermore, with respect to numerical ranges in this invention, it should be understood that each intermediate value between the upper and lower limits of the range is also specifically disclosed. Every smaller range between any stated value or intermediate value within a stated range, and any other stated value or intermediate value within said range, is also included in this invention. The upper and lower limits of these smaller ranges may be independently included or excluded from the range.

[0036] Unless otherwise stated, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. While only preferred methods and materials have been described herein, any methods and materials similar or equivalent to those described herein may be used in the implementation or testing of this invention. All references to this specification are incorporated by way of citation to disclose and describe methods and / or materials associated with those references. In the event of any conflict with any incorporated reference, the content of this specification shall prevail.

[0037] Various modifications and variations can be made to the specific embodiments described in this specification without departing from the scope or spirit of the invention, as will be apparent to those skilled in the art. Other embodiments derived from this specification will also be obvious to those skilled in the art. This specification and embodiments are merely exemplary.

[0038] The terms “include,” “including,” “have,” “contain,” etc., used in this article are all open-ended terms, meaning that they include but are not limited to.

[0039] Where specific techniques or conditions are not specified in the embodiments, they shall be performed in accordance with conventional techniques or conditions described in the literature in this field and the techniques or conditions recommended by the manufacturer. Reagents or instruments whose manufacturers are not specified are all commercially available conventional products. The source, trade name, and, where necessary, composition of the reagents used shall be indicated upon their first appearance; thereafter, unless otherwise specified, the same information shall apply to the same reagents used. The synthesis method of the all-hydroxy (extended) biphenyl[3] aromatic hydrocarbon can be found in the published article Fang, Z.-H.; Zhang, Z.-L.; Wang, R.-T.; Li, S.-H.; Lin, S.-J.; Zhou, Y.-R.; Chen, J.-Y.; Li, C.-J.; Meng, Q.-B. Efficient Reversal of Neuromuscular Blocking Agent-Induced Biological Functions and Side Effects by an Extended Biphen[3]arene Carboxylate. J. Med. Chem., 2024, 67, 21568-21576.

[0040] Example 1

[0041] Example 1 of this invention provides a method for synthesizing water-soluble (extended) biphenyl[3] aromatic derivatives, the specific steps of which are as follows:

[0042] Under nitrogen protection, 100 mg of perhydroxy(extended)biphenyl[3] aromatic hydrocarbon (0.11 mmol), 120 mg of sodium hydride (5.23 mmol), and 638 mg of propane sulfonate lactone (5.23 mmol) were placed in a round-bottom flask. 10 mL of a mixture of 1,4-dioxane and water (v:v = 1:1) was added for nucleophilic substitution reaction. After stirring under reflux at 90 °C for 24 hours, the organic solvent was removed by vacuum distillation. The remaining solution was placed in a dialysis bag with a molecular weight cutoff of 1000. The solution was stirred at room temperature in a beaker filled with water and the external solution was changed at fixed points every day. After 2 days, the solution in the dialysis bag was freeze-dried to obtain a water-soluble (extended)biphenyl[3] aromatic hydrocarbon derivative (yield: 42.83%). The product was verified by 1H NMR spectroscopy. The results are as follows: Figure 2 As shown.

[0043] Experimental Example 1

[0044] Experimental Example 1 of this invention uses the water-soluble (extended) biphenyl[3] aromatic derivative prepared in Example 1 to study the recognition behavior of capsaicin. The specific steps and results are as follows:

[0045] (1) Experimental equipment and reagents

[0046] The recognition behavior determines the ability of macrocyclic shielding capsaicin irritation. The nuclear magnetic resonance hydrogen spectrum (Bruker Ascend 600MHz) shows that the water-soluble (extended) biphenyl[3] aromatic derivative was synthesized in Example 1, and the tris(trimethylsilane) phosphate was purchased from Beijing Innocare Technology Co., Ltd.

[0047] (2) Experimental methods

[0048] Accurately weigh tris(trimethylsilane)phosphate to prepare a 5 mM heavy aqueous solution. Then, dissolve excess capsaicin, 1 mmol of water-soluble (extended) biphenyl[3] aromatic hydrocarbon, and 1 mmol of water-soluble (extended) biphenyl[3] aromatic hydrocarbon derivative containing excess capsaicin in 0.5 mL of the above-prepared heavy aqueous solution. Place the solution in a small centrifuge tube, heat and sonicate, filter using a filter head, and transfer to an NMR tube for analysis. The results are as follows: Figure 3 As shown.

[0049] (3) Experimental Results

[0050] like Figure 3 The NMR spectrum of the host-guest complex shown shows that, compared with the NMR signal of capsaicin alone, the chemical shifts of all hydrogen atoms of capsaicin after complexing with the water-soluble (extended) biphenyl[3] aromatic derivative shift to higher fields and exhibit a broadening effect. At the same time, due to the deshielding effect, the NMR signals of the water-soluble (extended) biphenyl[3] aromatic derivative shift to lower fields, indicating that there is a host-guest interaction between the two. In addition, using tris(trimethylsilane) phosphate as an internal standard, the solubility of capsaicin alone was calculated to be about 0.045 mM. When mixed with 2 mM of water-soluble (extended) biphenyl[3] aromatic derivative, the solubility reached 1.87 mM, which is nearly 50 times larger. This result indicates that the water-soluble (extended) biphenyl[3] aromatic derivative has a good solubilizing effect on capsaicin.

[0051] Experimental Example 2

[0052] In Experiment 2 of this invention, the cytotoxicity of the water-soluble (extended) biphenyl[3] aromatic derivative, capsaicin, and their complex prepared in Example 1 was evaluated. The specific steps and results are as follows:

[0053] (1) Experimental Samples

[0054] The water-soluble (extended) biphenyl[3] aromatic derivative was synthesized in Example 1. The human immortalized epidermal cells HaCaT were purchased from Beijing Zhong Sheng Aobang Biotechnology Co., Ltd., and the cell proliferation detection kit CCK-8 was purchased from Shanghai Dongren Chemical Technology Co., Ltd.

[0055] (2) Experimental methods

[0056] Human immortalized epidermal cells (HaCaT, Beijing Zhong Sheng Aobang Biotechnology Co., Ltd.) were cultured in DMEM medium containing 10% bovine serum peptide, 1% penicillin, and 1% streptomycin until stable passage. Cells in the logarithmic growth phase were seeded into 96-well plates (8000 cells / well) and incubated for 24 hours in an incubator (5% CO2, 37℃). Afterward, each well was replaced with 90 μL and 10 μL of fresh medium containing different concentrations of the test sample solution (640, 320, 160, 80, 40, 20, and 10 μM), with each concentration in 5 replicates. A control group was also included. After thorough mixing, the plates were incubated for another 24 hours. Cells were then stained with 10% CCK-8 medium (CCK-8 cell proliferation assay kit provided by Shanghai Dongren Chemical Technology Co., Ltd.). After a second 0.5 hours of incubation, the OD values ​​of each well were measured at 450 nm using an automated microplate reader. The results are shown below. Figure 4 , 5 As shown.

[0057] (3) Experimental Results

[0058] like Figure 4 As shown, the cytotoxicity of water-soluble (extended) biphenyl[3] aromatic derivatives on human immortalized epidermal cells HaCaT showed that even after co-incubation with the macrocycle at the highest concentration of 640 μM for 24 hours, the cell survival rate was still ≥98%, indicating that the macrocycle has low cytotoxicity. The cytotoxicity of capsaicin was then determined, and the results are as follows: Figure 5 As shown, HaCaT cells exhibit concentration-dependent cytotoxicity; however, when capsaicin is blended with an equivalent amount of water-soluble (extended) biphenyl[3] aromatic derivatives, the resulting cytotoxicity is significantly reduced. For example, at a dose of 640 μM, the cell survival rate increased from 2.15% to 84.62% after using macrocyclic compounds, demonstrating that the complexation with water-soluble (extended) biphenyl[3] aromatic derivatives can effectively inhibit the stimulation of capsaicin on cells.

[0059] Experimental Example 3

[0060] In Experiment 3 of this invention, the safety of the water-soluble (extended) biphenyl[3] aromatic derivative prepared in Example 1 was evaluated. The specific steps and results are as follows:

[0061] (1) Experimental Samples

[0062] Water-soluble (extended) biphenyl[3] aromatic hydrocarbons were synthesized in Example 1. Six-week-old Kunming mice were purchased from Beijing Spaford Biotechnology Co., Ltd.

[0063] (2) Experimental methods

[0064] Twelve six-week-old Kunming mice were purchased and housed normally for one week to acclimatize. They were then randomly divided into two groups (n=6 per group). The backs of the mice were shaved, and a 1cm diameter circular area was marked with a marker. One group received 20μL of double-distilled water, while the other group received 20μL of a 5mM macrocyclic aqueous solution evenly added to the marked area. Weight changes in each mouse were continuously observed and recorded over two weeks. After 14 days, the mice were euthanized by cervical dislocation, and their major organs were removed and weighed. Relevant organ indices were calculated. The results are as follows: Figure 6 , 7 As shown.

[0065] (3) Experimental Results

[0066] like Figure 6 The results showed changes in mouse weight. Compared with mice treated with double-distilled water, the experimental group mice showed a similar weight gain curve, and no obvious behavioral abnormalities were observed during this period. Organ index, as another indicator, reflects the systemic toxicity of water-soluble (extended) biphenyl[3] aromatic derivatives, such as Figure 7 As shown, there was no statistically significant difference in organ index between the experimental group mice and the blank control group after 14 days. The above results preliminarily indicate that water-soluble (extended) biphenyl[3] aromatic derivatives have good safety.

[0067] Test Example 4

[0068] Test Example 4 of this invention tested the water-soluble (extended) biphenyl[3] aromatic derivative prepared in Example 1 to alleviate capsaicin-induced scratching behavior in mice. The specific steps and results are as follows:

[0069] (1) Experimental Samples

[0070] Water-soluble (extended) biphenyl[3] aromatic hydrocarbons were synthesized in Example 1. Six-week-old Kunming mice were purchased from Beijing Spaford Biotechnology Co., Ltd., and dimethyl sulfoxide was purchased from Sinopharm Chemical Reagent Co., Ltd.

[0071] (2) Experimental methods

[0072] Twenty-four six-week-old Kunming mice were purchased and fed normally for one week to adapt to the environment. They were then randomly divided into four groups (n=6 per group). The backs of the mice were shaved and a circular area with a diameter of 1 cm was drawn with a marker. A 5 mM capsaicin sample was prepared using an aqueous solution containing 5% dimethyl sulfoxide. Then, 20 μL of capsaicin was evenly applied to the backs of each mouse in three of the groups. After air drying for 30 seconds, 20 μL of (extended) biphenyl[3] aromatic hydrocarbon aqueous solution was added to one group, and the other group was wiped with a cotton ball soaked in a macrocyclic aqueous solution. The number of scratches by each group of mice was counted in the next 5 minutes. The results are as follows: Figure 8 As shown.

[0073] (3) Experimental Results

[0074] Figure 8 The results of the test on the relief of capsaicin-induced scratching behavior in mice by water-soluble (extended) biphenyl[3] aromatic derivatives were shown. After applying capsaicin, mice exhibited high-frequency scratching behavior within 5 minutes, with an average of 22.7 times. In the group with an equivalent amount of macrocyclic aqueous solution, the number of scratches decreased to 8 times, indicating that subject-object recognition can effectively inhibit the irritation of capsaicin to the skin. After wiping with cotton balls soaked in macrocyclic aqueous solution, the average number of scratches in mice was only 5 times, indicating that water-soluble (extended) biphenyl[3] aromatic derivatives can be used as a scavenger to enrich capsaicin attached to the skin surface by solubilization.

[0075] The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the principle of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. A water-soluble extended biphenyl[3] aromatic derivative, characterized in that, The water-soluble extended biphenyl[3] aromatic derivative is an extended biphenyl[3] aromatic sulfonate, and the structure of the extended biphenyl[3] aromatic sulfonate is shown in Formula I: Formula I; The R is -CH2-CH2-CH2-SO3Na.

2. A method for preparing the water-soluble extended biphenyl[3] aromatic derivative as described in claim 1, characterized in that, Includes the following steps: The extended hydroxyl biphenyl[3] aromatic hydrocarbon was subjected to a nucleophilic substitution reaction with propane sulfonate lactone in a mixture of 1,4-dioxane and water in the presence of sodium hydride. The solution after the nucleation substitution reaction was completed was dialyzed and dried to obtain extended biphenyl[3] aromatic hydrocarbon sulfonate.

3. The preparation method according to claim 2, characterized in that, The molar and volume ratio of the mixture of the fully hydroxyl extended biphenyl[3] aromatic hydrocarbon, sodium hydride, propane sulfonate lactone, 1,4-dioxane and water is 0.1~0.2 mmol:4~6 mmol:4~6 mmol:8~10 mL.

4. The preparation method according to claim 3, characterized in that, In the mixture of 1,4-dioxane and water, the volume ratio of 1,4-dioxane to water is 1~2:1~2.

5. The preparation method according to claim 2, characterized in that, The nucleophilic substitution reaction is carried out at a temperature of 85-95°C for a time of 12-36 hours.

6. The preparation method according to claim 2, characterized in that, The dialysis method is as follows: Place the solution in a dialysis bag, place the dialysis bag in water and stir. After dialysis, take the solution from the dialysis bag for the next step of processing.

7. The preparation method according to claim 6, characterized in that, The molecular weight cutoff of the dialysis bag is 900-1100; the dialysis time is 1.5-3 days.

8. The use of a water-soluble extended biphenyl[3] aromatic derivative as described in claim 1 in the preparation of a drug to relieve the irritation caused by capsaicin.

9. The application in the preparation of a medicament for relieving irritation caused by capsaicin according to claim 8, characterized in that, This includes applying a solution of extended biphenyl[3] aromatic hydrocarbon derivatives to the exposed areas of capsaicin to inhibit its irritation through pharmacokinetic means; or using extended biphenyl[3] aromatic hydrocarbon derivatives as a decontaminant to remove capsaicin adhering to the skin by solubilization and wiping. The benign solvent for the extended biphenyl[3] aromatic derivative is one or a mixture of water, phosphate buffer, dimethyl sulfoxide, and methanol.

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