Glucose substituted pyrazole compound and borneol composition and preparation method thereof

Abstract

The invention discloses a glucose-substituted pyrazole compound and borneol composition and a preparation method and application thereof, and relates to the technical field of pharmaceutical compositions.The composition is endowed with highly-accurate tumor targeted release characteristics through a dynamic boric acid ester bond cross-linked structure; specific high-concentration cis-vicinal diol substances in tumor cells can trigger boric acid ester bond exchange reaction to promote the composition structure to disintegrate and release the medicine, and the medicine release amount in a normal cell environment is extremely small, so that the medicine can act on tumor tissues in a concentrated manner, the medicine concentration of tumor parts is remarkably improved, and the curative effect of the medicine is improved. Meanwhile, the distribution of the medicine in normal tissues is greatly reduced, the toxic and side effects on the normal cells are reduced, the safety and tolerance in the treatment process are improved, and the improvement of the overall effect of tumor treatment and the prognosis of patients is assisted.

Description

Technical Field

[0001] The present invention relates to the technical field of pharmaceutical compositions, in particular to a composition of a glucose-substituted pyrazole compound and borneol, and a preparation method and application thereof. Background Art

[0002] Tumors pose a serious threat to human health. Seeking efficient and safe tumor treatment drugs has always been a research focus in the medical field. The precise release and targeting of drugs at the tumor site are key links to improving treatment effects and reducing toxic side effects. Therefore, there is an urgent clinical need to develop drug compositions with special response mechanisms that can adapt to the tumor microenvironment.

[0003] However, existing tumor therapeutic drug compositions generally lack a specific response mechanism to the tumor microenvironment. Regarding the research on achieving controlled drug release through chemical bonding, existing technologies mostly focus on simple combinations of conventional chemical bonds and lack designs that use dynamic covalent bonds to construct responsive structures. In addition, most drug release relies on passive diffusion, which not only leads to ineffective distribution of drugs in non-tumor sites and increases toxic side effects on normal cells, but also affects efficacy due to insufficient drug concentration in tumor sites. It also fails to fully utilize the high concentration of cis-vicinal diols in tumor cells, resulting in poor accuracy and controllability of drug release.

[0004] In summary, it is urgent to develop a composition that can accurately respond to the tumor microenvironment and achieve drug-specific release. The present invention is based on a dynamic boronate bond cross-linking structure, which gives the composition the characteristic of "tumor microenvironment-triggered release", which is used to solve the problems of poor drug targeting and large toxic side effects in the existing technology, and provides new ideas for the development of tumor treatment drugs. Summary of the Invention

[0005] The purpose of the present invention is to make up for the shortcomings of the existing technology and provide a composition of a glucose-substituted pyrazole compound and borneol, as well as its preparation method and application. It can construct a tumor microenvironment-specific response system through dynamic boronate bonds. The high concentration of cis-vicinal diol substances in tumor cells can trigger the boronate bond exchange reaction, causing the structure of the composition to disintegrate and release drugs. Normal cells have a low content of cis-vicinal diols and release very little drug. This design greatly improves the accuracy of the drug's effect on tumor cells and reduces interference with normal cells. It can not only increase the drug concentration in the tumor site and enhance the therapeutic effect, but also reduce systemic toxicity, so that patients can better tolerate treatment.

[0006] In order to solve the above technical problems, the present invention provides the following technical solutions: a composition of a glucose-substituted pyrazole compound and borneol, a preparation method and application thereof, wherein the composition is an injection solution composed of a supramolecular inclusion compound, an injection carrier and basic excipients;

[0007] The supramolecular inclusion compound is formed by combining the compound of formula I' and the borneol derivative of formula II through stereo complementarity and intermolecular forces;

[0008] The injection carrier comprises a supramolecular inclusion compound, a cosolvent, and a stabilizer;

[0009] The compound of formula I' is composed of 1-phenyl-3-methyl-5-OD-glucoside-pyrazole having a structure of formula I, wherein the C4 position thereof is modified with a two-photon absorption group naphthalene imide connected via an amide bond. The structural formula I of the 1-phenyl-3-methyl-5-OD-glucoside-pyrazole is:

[0010]

[0011] Furthermore, the I' compound is C6H5-N(CH3)-NC(glucose)-NH-CO-naphthalimide, wherein the β-D configuration of the glucose group is covalently linked to the naphthalimide group via an amide bond, and the 3-hydroxyl group of the β-D of the glucose group retains reactivity. The naphthalimide group also has a two-photon absorption function, providing a structural basis for deep tissue targeted drug release. The structural formula I' of the I' compound is:

[0012]

[0013] Furthermore, the hydroxyl group of the borneol molecule in the borneol derivative of formula II is coupled to the 3-hydroxyl group of the glucose group in the compound of formula I' through an o-nitrobenzyl photosensitive linker to form a conjugate of formula II. The o-nitrobenzyl group acts as a photocleavable group, and the nitrobenzyl ring fragmentation reaction occurs under two-photon excitation, specifically releasing the borneol molecule. The structural formula of formula II is:

[0014]

[0015] Furthermore, the stabilizer in the injection carrier is 2% of a lyoprotectant and 0.08% of an osmotic pressure regulator, with a content of 1-3%, and the cosolvent is propylene glycol;

[0016] The stabilizer and propylene glycol work together to ensure the stability and biocompatibility of the preparation, and release borneol through two-photon excitation to act on targeted therapy.

[0017] Furthermore, the concentration of the supramolecular inclusion complex in the injection carrier is 0.1-5.0 mg / mL.

[0018] Furthermore, the basic auxiliary materials include: pH regulator, antioxidant, eluent, catalyst;

[0019] The pH regulator is sodium hydroxide, which is used to adjust the pH value of the injection to 7.0-7.8;

[0020] The antioxidant is sodium bisulfite with a concentration of 0.05-0.1%;

[0021] The eluent is chloroform and methanol in a molar ratio of 8:1;

[0022] The catalyst is copper sulfate CuSO4 and sodium ascorbate in a molar ratio of 1:2.

[0023] Furthermore, the injection of the composition is used in the following fields:

[0024] Treatment of brain diseases: Borneol is delivered through the blood-brain barrier, and the inclusion complex is released at the site of cerebral ischemia and neuroinflammation by triggering near-infrared light, which is used to treat stroke and Alzheimer's disease;

[0025] Precision tumor treatment: Two-photon excitation enables light-controlled drug release in deep tumor tissues, suitable for gliomas and breast cancer brain metastases;

[0026] Real-time therapeutic monitoring: The naphthalene imide group provides fluorescent tracing function, which can monitor drug distribution and release kinetics through in vivo imaging.

[0027] On the other hand, a method for preparing a composition of a glucose-substituted pyrazole compound and borneol is provided, wherein the specific steps of the method are:

[0028] S100, Synthesis of the Compound of Formula I': Dissolve the compound of Formula I, 1-phenyl-3-methyl-5-OD-glucoside-pyrazole, and naphthalene imide carboxylic acid in anhydrous DMF at a molar ratio of 1:1.2, add 1.5% EDC·HCl and 1.2% NHS, and react at 25°C under nitrogen for 12 hours. After purification by adding an eluent, collect the eluate to obtain a white solid, which is the compound of Formula I';

[0029] S200, preparation of a borneol derivative of formula II: dissolving borneol and o-nitrobenzyl bromide in acetone at a molar ratio of 1:1.5, adding potassium carbonate, and reacting at reflux at 60°C for 6 hours to produce an o-nitrobenzyl-protected borneol derivative, mixing the derivative with a compound of formula I' at a molar ratio of 1:1, adding a catalyst, stirring and reacting for 12 hours, collecting and filtering the precipitated solid, which is the borneol derivative of formula II;

[0030] S300, supramolecular inclusion: Formula I' and Formula II were dissolved in propylene glycol solvent at a molar ratio of 1:1.2, and stirred under ultrasound assistance for 6 hours to induce inclusion complex formation. The solution was transferred to a dialysis bag and dialyzed in deionized water for 24 hours to remove unincluded substances, and the solution was freeze-dried to obtain a supramolecular inclusion complex as a white powder;

[0031] S400, preparation of injection: adding a stabilizer to the inclusion compound solution, adjusting the pH to 7.5±0.2 with sodium hydroxide, adding physiological saline to the final volume, sterilizing and filtering through a filter membrane, and then filling into an ampoule to obtain the injection of the composition of the glucose-substituted pyrazole compound and borneol;

[0032] S500, verification of light-controlled release: The injection solution was pre-frozen to -80°C and then freeze-dried to obtain a white freeze-dried powder, which was reconstituted with water for injection before use. The borneol release efficiency was verified by two-photon laser.

[0033] Compared with the prior art, the composition of a glucose-substituted pyrazole compound and borneol, its preparation method and application have the following beneficial effects:

[0034] 1. The present invention imparts highly precise tumor-targeted release properties to the composition through a dynamic boronate cross-linking structure. The high concentration of cis-vicinal diol substances unique to tumor cells can trigger a boronate exchange reaction, causing the composition structure to disintegrate and release the drug, while the amount of drug released in a normal cell environment is extremely small. This property enables the drug to act concentratedly on tumor tissue, significantly increasing the drug concentration at the tumor site and enhancing the killing effect on tumor cells. At the same time, it greatly reduces the distribution of the drug in normal tissues, reduces the toxic side effects on normal cells, improves the safety and tolerability of the treatment process, and helps improve the overall effect of tumor treatment and patient prognosis.

[0035] 2. The dynamic boronate cross-linking design of the present invention provides a flexible and controllable regulation mechanism for drug release. The concentration difference of cis-vicinal diol substances in the tumor microenvironment can regulate the drug release rate in real time, so that the drug release is matched with the tumor development stage, facilitating personalized treatment. In addition, this dynamic regulation avoids the sudden release of the drug, ensures that the drug concentration remains in the effective range for a long time, prolongs the drug action time, reduces the number of dosing times, and improves the convenience and compliance of patients.

[0036] Other advantages, objects and features of the present invention will be described in part in the following description and, in part, will be apparent to those skilled in the art based on an examination of the following or may be learned from the practice of the invention. BRIEF DESCRIPTION OF THE DRAWINGS

[0037] To more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the following briefly introduces the drawings required for use in the embodiments or the description of the prior art. Obviously, the drawings described below are only some embodiments of the present invention. Those skilled in the art can also derive other drawings based on these drawings without inventive effort.

[0038] Figure 1 The present invention is a flow chart of a method for preparing a composition of a glucose-substituted pyrazole compound and borneol. DETAILED DESCRIPTION

[0039] In order to further illustrate the technical means and effects adopted by the present invention to achieve the predetermined purpose of the invention, the specific implementation methods, structures, features and effects of the present invention are described in detail below in conjunction with the accompanying drawings and preferred embodiments.

[0040] The present invention provides a composition of a glucose-substituted pyrazole compound and borneol, which is an injection solution consisting of a supramolecular inclusion compound, an injection carrier and basic auxiliary materials;

[0041] The supramolecular inclusion compound is formed by combining the compound of formula I' and the borneol derivative of formula II through stereo complementarity and intermolecular forces;

[0042] The injection carrier comprises a supramolecular inclusion compound, a cosolvent, and a stabilizer, wherein the stabilizer is 2% of a lyoprotectant and 0.08% of an osmotic pressure regulator, with a content of 1-3%. The cosolvent is propylene glycol. The stabilizer and propylene glycol synergistically ensure the stability and biocompatibility of the preparation, release borneol through two-photon excitation, and act on targeted therapy. The concentration of the supramolecular inclusion compound is 0.1-5.0 mg / mL.

[0043] The compound of formula I' is composed of 1-phenyl-3-methyl-5-OD-glucoside-pyrazole having a structure of formula I, wherein the C4 position thereof is modified with a two-photon absorption group naphthalene imide connected via an amide bond. In addition, the compound I' is C6H5-N(CH3)-NC(glucose)-NH-CO-naphthalene imide, wherein the β-D configuration of the glucose group is covalently linked to the naphthalene imide group via an amide bond, and the 3-position hydroxyl group of the β-D of the glucose group retains reaction activity. The naphthalene imide group also has a two-photon absorption function, providing a structural basis for deep tissue targeted drug release.

[0044] The hydroxyl group of the borneol molecule in the borneol derivative of the formula II is coupled to the 3-hydroxyl group of the glucose group in the compound of the formula I' via an o-nitrobenzyl photosensitive linker to form a conjugate of the formula II. The o-nitrobenzyl group acts as a photocleavable group, and the nitrobenzyl ring undergoes a fragmentation reaction under two-photon excitation, specifically releasing the borneol molecule.

[0045] The basic auxiliary materials include: pH regulator, antioxidant, eluent, catalyst;

[0046] The pH regulator is sodium hydroxide, which is used to adjust the pH value of the injection to 7.0-7.8;

[0047] The antioxidant is sodium bisulfite with a concentration of 0.05-0.1%;

[0048] The eluent is chloroform and methanol in a molar ratio of 8:1;

[0049] The catalyst is copper sulfate (CuSO4) and sodium ascorbate in a molar ratio of 1:2.

[0050] The preparation method of the composition based on the glucose-substituted pyrazole compound and borneol is as follows: Figure 1 As shown, the specific steps of the preparation method of the composition are:

[0051] S100, Synthesis of the Compound of Formula I': Dissolve the compound of Formula I, 1-phenyl-3-methyl-5-OD-glucoside-pyrazole, and naphthalene imide carboxylic acid in anhydrous DMF at a molar ratio of 1:1.2, add 1.5% EDC·HCl and 1.2% NHS, and react at 25°C under nitrogen for 12 hours. After purification by adding an eluent, collect the eluate to obtain a white solid, which is the compound of Formula I';

[0052] S200, preparation of a borneol derivative of formula II: dissolving borneol and o-nitrobenzyl bromide in acetone at a molar ratio of 1:1.5, adding potassium carbonate, and reacting at reflux at 60°C for 6 hours to produce an o-nitrobenzyl-protected borneol derivative, mixing the derivative with a compound of formula I' at a molar ratio of 1:1, adding a catalyst, stirring and reacting for 12 hours, collecting and filtering the precipitated solid, which is the borneol derivative of formula II;

[0053] S300, supramolecular inclusion: dissolving Formula I′ and Formula II in a propylene glycol solvent at a molar ratio of 1:1.2, stirring under ultrasound assistance for 6 hours to induce inclusion complex formation, transferring the solution to a dialysis bag, dialyzing in deionized water for 24 hours to remove unincluded substances, and freeze-drying the solution to obtain a supramolecular inclusion complex;

[0054] S400, preparation of injection: add a stabilizer to the inclusion compound solution, adjust the pH to 7.5±0.2 with sodium hydroxide, add physiological saline to the final volume, sterilize and filter through a filter membrane, and then fill into an ampoule to obtain the injection of the glucose-substituted pyrazole compound and borneol combination:

[0055] S500, verification of light-controlled release: The injection solution was pre-frozen to -80°C and then freeze-dried to obtain a white freeze-dried powder, which was reconstituted with water for injection before use. The borneol release efficiency was verified by two-photon laser.

[0056] Example 1

[0057] Synthesis of the compound of formula I': Accurately weigh 1.0 g of 1-phenyl-3-methyl-5-OD-glucoside-pyrazole and 1.5 g of naphthalene imide carboxylic acid, dissolve the two in 10 mL of anhydrous DMF, and in a fume hood, add 1.5% EDC·HCl and 1.2% NHS in sequence. Purify the mixture by stirring in an oil bath at 25°C for 12 hours. After the reaction, chloroform and methanol in a molar ratio of 8:1 are added as eluents for purification by silica gel column chromatography. The eluate is collected and concentrated under reduced pressure to obtain a white solid, namely the compound of formula I', weighing 1.3 g, with a yield of approximately 72%.

[0058] Preparation of borneol derivatives of formula II: Weigh 1.2 g of borneol and 1.8 g of o-nitrobenzyl bromide, dissolve them in 15 mL of acetone, add 1.1 g of potassium carbonate, and reflux in an oil bath at 60°C for 6 hours. After the reaction is completed, cool to room temperature, filter out the potassium carbonate solid, and mix the obtained o-nitrobenzyl-protected borneol derivative with 1.3 g of the compound of formula I′ in a 1:1 molar ratio. Add copper sulfate CuSO4 and sodium ascorbate in a molar ratio of 1:2 as a catalyst, and stir the reaction for 12 hours. After the reaction is completed, filter the precipitated solid, wash it with a small amount of acetone, and dry it to obtain 1.5 g of a borneol derivative of formula II with a yield of about 75%.

[0059] Supramolecular inclusion: 1.0 g of the compound of formula I′ and 1.2 g of the compound of formula II were dissolved in 10 mL of propylene glycol solvent. The reaction vessel was placed in an ultrasonic cleaner and stirred under ultrasonic assistance for 6 hours to induce inclusion complex formation. The ultrasonic power was set to 300 W and the frequency was 40 kHz. After the reaction, the solution was transferred to a dialysis bag with a molecular weight cutoff of 3500 Da and dialyzed in deionized water for 24 hours. The deionized water was replaced every 4 hours to remove unincluded substances. After the dialysis, the solution was freeze-dried to obtain 1.4 g of a white powdery supramolecular inclusion complex with a yield of approximately 70%.

[0060] Preparation of injection: Weigh 0.5 g of the supramolecular inclusion complex, add an appropriate amount of water for injection to dissolve it, then add 2% of a lyoprotectant and 0.08% of an osmotic pressure regulator as stabilizers, carefully adjust the pH to 7.5 with sodium hydroxide solution, add physiological saline to a final volume of 50 mL, stir evenly, sterile filter through a 0.22 μm filter membrane, and aseptically fill into ampoules, 2 mL per bottle, to obtain the injection of the composition of the glucose-substituted pyrazole compound and borneol.

[0061] Verification of light-controlled release: Take 1 mL of the above injection solution in a penicillin bottle, pre-freeze it to -80°C and then freeze-dry it to obtain a white freeze-dried powder. Before use, reconstitute it with 1 mL of water for injection, use two-photon laser irradiation, and use high-performance liquid chromatography to determine the release amount of borneol at different time points. The results showed that after 10 minutes of irradiation, the borneol release efficiency reached 30%, after 30 minutes of irradiation, the release efficiency was 60%, and after 60 minutes of irradiation, the release efficiency stabilized at about 85%.

[0062] Implementation effect: In a cell model experiment simulating cerebral ischemia, after the cells were treated with the injection of the composition for 24 hours, the cell survival rate reached 80%, and the secretion of the inflammatory factor IL-6 was reduced by 40% compared with the untreated group. In the killing experiment of glioma cells, the proliferation inhibition rate of tumor cells reached 70% after 48 hours of drug action. Through in vivo imaging observation, the fluorescence signal of the naphthaleneimide group was stable, which can accurately mark the distribution location of the drug in the body, and the fluorescence intensity at the tumor site maintained a steady increase within 6 hours after administration, providing a reliable basis for real-time monitoring of the drug treatment process.

[0063] Example 2

[0064] Synthesis of the compound of formula I': 1.2 g of 1-phenyl-3-methyl-5-OD-glucoside-pyrazole and 1.8 g of naphthalene imide carboxylic acid were weighed and dissolved in 12 mL of anhydrous DMF. 1.5% EDC·HCl and 1.2% NHS were added, and nitrogen was passed through for 5 minutes. The reaction was carried out at 25°C for 12 hours. After the reaction, the mixture was purified with an eluent, and the eluate was collected and concentrated under reduced pressure to obtain 1.6 g of the compound of formula I' as a white solid in a yield of approximately 75%.

[0065] Preparation of borneol derivatives of formula II: Weigh 1.5 g of borneol and 2.2 g of o-nitrobenzyl bromide, dissolve them in 18 mL of acetone, add 1.3 g of potassium carbonate, reflux at 60°C for 6 hours, cool and filter, mix the o-nitrobenzyl-protected borneol derivative with 1.6 g of the compound of formula I′, add a catalyst, stir and react for 12 hours, filter, wash, and dry to obtain 1.9 g of a borneol derivative of formula II with a yield of about 78%.

[0066] Supramolecular inclusion complex: 1.2 g of the compound of formula I′ and 1.4 g of the compound of formula II were dissolved in 12 mL of propylene glycol and stirred with ultrasonic power of 350 W and frequency of 45 kHz for 6 hours. The mixture was dialyzed and freeze-dried to obtain 1.7 g of supramolecular inclusion complex with a yield of about 72%.

[0067] Preparation of injection: Weigh 0.6 g of supramolecular inclusion complex, dissolve it in water for injection, add stabilizer, adjust the pH to 7.5 with sodium hydroxide, add normal saline to 60 mL, filter and sterilize, and then fill into ampoule bottles, 2 mL per bottle.

[0068] Light-controlled release verification: Light-controlled release verification was carried out according to the method of Example 1. Under two-photon laser irradiation, the borneol release efficiency was 32% after 10 minutes of irradiation; 63% after 30 minutes; and stabilized at about 88% after 60 minutes.

[0069] Results: In a cell model simulating cerebral ischemia, 24 hours after treatment with the combination, the cell survival rate reached 82%, and the secretion of the inflammatory factor IL-6 was reduced by 42% compared to the untreated group. In a glioma cell killing experiment, the proliferation inhibition rate reached 72% after 48 hours of drug exposure. In vivo imaging showed that the fluorescence signal of the naphthaleneimide group was stable, clearly demonstrating the drug's distribution path in vivo. The fluorescence intensity at the tumor site showed a significant increase within 6 hours of administration, facilitating precise monitoring of the progress of drug treatment.

[0070] Example 3

[0071] Synthesis of the compound of formula I': 1.5 g of 1-phenyl-3-methyl-5-OD-glucoside-pyrazole and 2.2 g of naphthalene imide carboxylic acid were dissolved in 15 mL of anhydrous DMF, and 1.5% EDC·HCl and 1.2% NHS were added. The mixture was purged with nitrogen and reacted at 25°C for 12 hours. The mixture was purified with an eluent and concentrated under reduced pressure to obtain 2.0 g of the compound of formula I' in a yield of approximately 77%.

[0072] Preparation of borneol derivatives of formula II: Weigh 1.8 g of borneol and 2.7 g of o-nitrobenzyl bromide, dissolve in 20 mL of acetone, add 1.6 g of potassium carbonate, and reflux at 60°C for 6 hours. Subsequently, react and treat with the compound of formula I′ to obtain 2.3 g of the borneol derivative of formula II with a yield of approximately 80%.

[0073] Supramolecular inclusion complex: 1.5 g of the compound of formula I′ and 1.8 g of the compound of formula II were dissolved in 15 mL of propylene glycol and stirred with ultrasonic power of 400 W and frequency of 50 kHz for 6 hours. The mixture was dialyzed and freeze-dried to obtain 2.1 g of supramolecular inclusion complex with a yield of about 73%.

[0074] Preparation of injection: Weigh 0.8 g of supramolecular inclusion complex, dissolve in water for injection, add stabilizer, adjust pH to 7.5, add normal saline to 80 mL, filter and sterilize, and bottle, 2 mL per bottle.

[0075] Light-controlled release verification: The light-controlled release verification method was the same as that in Example 1. Under the same two-photon laser irradiation conditions, the borneol release efficiency was 35% at 10 minutes, 65% at 30 minutes, and about 90% at 60 minutes.

[0076] Implementation effect: In a cell model experiment simulating cerebral ischemia, after 24 hours of treatment with the injection of the composition, the cell survival rate reached 84%, and the secretion of the inflammatory factor IL-6 was reduced by 45% compared with the untreated group. The killing experiment on glioma cells showed that after 48 hours of drug action, the proliferation inhibition rate of tumor cells reached 75%. In in vivo imaging, the fluorescence signal of the naphthaleneimide group was strong and stable, which could clearly show the dynamic distribution of the drug in the body. The fluorescence intensity at the tumor site increased significantly within 6 hours after administration, providing intuitive data for the evaluation of drug efficacy.

[0077] Example 4

[0078] Synthesis of the compound of formula I': Weigh 1.8 g of 1-phenyl-3-methyl-5-OD-glucoside-pyrazole, 2.6 g of naphthalene imide carboxylic acid, and 0.12 g of NHS, purge with nitrogen, and react at 25°C for 12 hours. After purification, 2.4 g of the compound of formula I' was obtained with a yield of approximately 79%.

[0079] Preparation of Borneol Derivative of Formula II: Weigh 2.1 g of borneol and 3.1 g of o-nitrobenzyl bromide, dissolve in 22 mL of acetone, add 1.9 g of potassium carbonate, and reflux at 60°C for 6 hours. Subsequent operation yields 2.7 g of Borneol Derivative of Formula II, with a yield of approximately 82%.

[0080] Supramolecular inclusion complex: 1.8 g of the compound of formula I′ and 2.2 g of the compound of formula II were dissolved in 18 mL of propylene glycol, and stirred with ultrasonic power of 450 W and frequency of 55 kHz for 6 hours. The mixture was dialyzed and freeze-dried to obtain 2.5 g of supramolecular inclusion complex with a yield of about 75%.

[0081] Preparation of injection: Weigh 1.0 g of supramolecular inclusion complex, dissolve in water for injection, add stabilizer, adjust pH to 7.5, add normal saline to 100 mL, filter and sterilize, and bottle, 2 mL per bottle.

[0082] Light-controlled release verification: According to the light-controlled release verification process of Example 1, under the same laser conditions, the borneol release efficiency is 38% in 10 minutes, 68% in 30 minutes, and about 92% in 60 minutes.

[0083] Results: In a cell model simulating cerebral ischemia, 24 hours after treatment with the combination, the cell survival rate reached 86%, and the secretion of the inflammatory factor IL-6 was reduced by 48% compared to the untreated group. In a glioma cell killing experiment, the tumor cell proliferation inhibition rate reached 78% after 48 hours of drug exposure. In vivo imaging revealed that the naphthaleneimide group's fluorescence signal was strong, clear, and stable, enabling accurate tracking of the drug's distribution within the body. The fluorescence intensity at the tumor site continued to increase steadily within 6 hours after administration, facilitating timely monitoring of the drug's therapeutic effects.

[0084] Example 5

[0085] Synthesis of the compound of formula I': 2.0 g of 1-phenyl-3-methyl-5-OD-glucoside-pyrazole and 2.9 g of naphthalene imide carboxylic acid were dissolved in 20 mL of anhydrous DMF, and 1.5% EDC·HCl and 1.2% NHS were added. The mixture was purged with nitrogen and reacted at 25°C for 12 hours. After treatment, 2.7 g of the compound of formula I' was obtained with a yield of approximately 80%.

[0086] Preparation of borneol derivatives of formula II: Weigh 2.4 g of borneol and 3.6 g of o-nitrobenzyl bromide, dissolve in 25 mL of acetone, add 2.2 g of potassium carbonate, and reflux at 60°C for 6 hours. Subsequent treatment gives 3.1 g of borneol derivatives of formula II with a yield of about 83%.

[0087] Supramolecular inclusion complex: 2.0 g of the compound of formula I′ and 2.4 g of the compound of formula II were dissolved in 20 mL of propylene glycol, and stirred with ultrasonic power of 500 W and frequency of 60 kHz for 6 hours. The mixture was dialyzed and freeze-dried to obtain 2.8 g of supramolecular inclusion complex with a yield of about 76%.

[0088] Preparation of injection: Weigh 1.2 g of supramolecular inclusion complex, dissolve in water for injection, add stabilizer, adjust pH to 7.5, add normal saline to 120 mL, filter and sterilize, and bottle, 2 mL per bottle.

[0089] Light-controlled release verification: Using the same light-controlled release verification method as in Example 1, under the same two-photon laser irradiation, the borneol release efficiency was 40% at 10 minutes, 70% at 30 minutes, and about 93% at 60 minutes.

[0090] Implementation results: In a cell model simulating cerebral ischemia, 24 hours after treatment with the compound, the cell survival rate reached 88%, and the secretion of the inflammatory factor IL-6 was reduced by 50% compared to the untreated group. In a glioma cell killing experiment, the drug inhibited tumor cell proliferation by 80% after 48 hours of treatment. In vivo imaging results showed that the fluorescence signal of the naphthaleneimide group was stable, clearly showing the distribution details of the drug in the body. The fluorescence intensity at the tumor site increased significantly within 6 hours after administration, providing strong support for the accurate assessment of drug efficacy.

[0091] Comparative Example 1

[0092] Preparation of a composition without a dynamic borate ester bond: Prepare the compound of formula I′ using the same method as in Example 1, directly mixing borneol with the compound of formula I′ in a molar ratio of 1:1.2 without coupling the o-nitrobenzyl photosensitive linker. Dissolve the mixture in propylene glycol, ultrasonically stir for 6 hours, dialyze, and lyophilize to obtain a powder.

[0093] Preparation and verification of the injection solution: Weigh 0.5 g of the powder, dissolve it in water for injection, add a stabilizer, adjust the pH to 7.5, add physiological saline to 50 mL, filter and sterilize, and bottle it for light-controlled release verification. After 60 minutes of two-photon laser irradiation, almost no borneol is released. In the tumor cell experiment, compared with the composition of Example 1, the killing effect on tumor cells is significantly reduced, but there is no significant difference in toxicity to normal cells.

[0094] Results: In a cell model simulating cerebral ischemia, 24 hours after treatment with the composition, cell survival was reduced to just 50%, and secretion of the inflammatory factor IL-6 decreased by 10% compared to the untreated group. In a glioma cell killing experiment, 48 hours after treatment, tumor cell proliferation inhibition was only 30%. During in vivo imaging, the lack of an effective fluorescent tracing mechanism prevented clear visualization of the drug's distribution within the body, making it difficult to effectively monitor the therapeutic process.

[0095] Comparative Example 2

[0096] Preparation of a composition replacing key raw materials: a common fluorescent group was used to replace the naphthalene imide group to modify the compound of formula I, and a similar compound of formula I′ was synthesized. Subsequently, the borneol derivative, supramolecular inclusion compound and injection of formula II were prepared according to the method of Example 1.

[0097] Performance Verification: Light-controlled release verification was performed, and the release efficiency was similar to that of Example 1. However, in the real-time treatment monitoring experiment, it was found through in vivo imaging that the fluorescent tracing effect was far inferior to the composition in Example 1, and it was impossible to clearly monitor the drug distribution and release kinetics, which affected the evaluation of the treatment effect.

[0098] Implementation results: In a cell model simulating cerebral ischemia, 24 hours after treatment with the composition, the cell survival rate was 70%, and the secretion of the inflammatory factor IL-6 was reduced by 30% compared to the untreated group. In a glioma cell killing experiment, the drug inhibited tumor cell proliferation by 60% after 48 hours of treatment. Although the approximate distribution of the drug can be observed through fluorescence, the unstable and low-resolution fluorescence signal makes it difficult to accurately determine the drug's accumulation and release dynamics at the tumor site, resulting in significant errors in the assessment of therapeutic efficacy.

[0099] The implementation effects of the above embodiments and comparative examples are shown in the following table:

[0100]

[0101] In summary, through comparative analysis of the experimental data of multiple examples and comparative examples, it can be seen that the composition of the glucose-substituted pyrazole compound and borneol of the present invention exhibits significant advantages in various performance indicators, among which:

[0102] In terms of cell protection and inflammation regulation, the survival rate of simulated cerebral ischemia cells in the examples gradually increased with experimental optimization, from 80% in Example 1 to 88% in Example 5, and the reduction in the secretion of the inflammatory factor IL-6 also increased from 40% to 50%, indicating that the composition has a good protective effect on cerebral ischemia damage and can effectively inhibit the inflammatory response. In terms of tumor treatment effect, the glioma cell proliferation inhibition rate of the examples continued to increase, reaching 80% in Example 5, which strongly demonstrated its efficient killing ability against tumor cells.

[0103] The in vivo imaging fluorescence effect and the change in fluorescence intensity at the tumor site are the highlights of the present invention. The fluorescent signal provided by the naphthalene imide group in the embodiment is clear, stable and bright, which can not only accurately mark and track drug distribution, but also show the details and dynamic changes of drug distribution. The fluorescence intensity at the tumor site continues to grow steadily within 6 hours after administration, providing a reliable basis for real-time monitoring of drug treatment progress and evaluation of efficacy.

[0104] In comparison, Comparative Example 1 lacks a dynamic boronate bond, so borneol cannot be effectively released, and performs extremely poorly in terms of cell protection, tumor killing, and fluorescence tracing; although Comparative Example 2 has the ability to release drugs under light control, the ordinary fluorescent group results in poor fluorescence tracing effect, making it difficult to accurately evaluate the therapeutic effect.

[0105] Overall, the dynamic boronate cross-linking design of the present invention provides a flexible and controllable regulatory mechanism for drug release. The concentration difference of cis-vicinal diol substances in the tumor microenvironment can regulate the drug release rate in real time, so that the drug release is matched with the tumor development stage, facilitating personalized treatment.

[0106] The above description is merely a preferred embodiment of the present invention and does not constitute any form of limitation to the present invention. Although the present invention has been disclosed as above in terms of a preferred embodiment, it is not intended to limit the present invention. Any person skilled in the art can, without departing from the scope of the technical solution of the present invention, make some changes or modifications to equivalent embodiments using the technical contents disclosed above. However, any brief modifications, equivalent changes and modifications made to the above embodiments based on the technical essence of the present invention without departing from the content of the technical solution of the present invention are still within the scope of the technical solution of the present invention.

Claims

1. A composition of a glucose-substituted pyrazole compound and borneol, characterized in that: The composition is an injection liquid consisting of a supramolecular inclusion compound, an injection carrier and basic auxiliary materials; The supramolecular inclusion compound is formed by combining the compound of formula I' and the borneol derivative of formula II through stereo complementarity and intermolecular forces; The injection carrier comprises a supramolecular inclusion compound, a cosolvent, and a stabilizer; The compound of formula I′ is composed of 1-phenyl-3-methyl-5-OD-glucoside-pyrazole having a structure of formula I, wherein the C4 position thereof is modified with a two-photon absorption group naphthalene imide connected via an amide bond. The structural formula I of the 1-phenyl-3-methyl-5-OD-glucoside-pyrazole is:

2. The composition of a glucose-substituted pyrazole compound and borneol according to claim 1, characterized in that: The compound I' is C6H5-N(CH3)-NC(glucose)-NH-CO-naphthalimide, wherein the β-D configuration of the glucose group is covalently linked to the naphthalimide group via an amide bond, and the 3-hydroxyl group of the β-D of the glucose group retains reactivity. The naphthalimide group also has a two-photon absorption function, providing a structural basis for deep tissue targeted drug release. The structural formula I' of the compound I' is:

3. The composition of a glucose-substituted pyrazole compound and borneol according to claim 1, characterized in that: The hydroxyl group of the borneol molecule in the borneol derivative of formula II is coupled to the 3-hydroxyl group of the glucose group in the compound of formula I' through an o-nitrobenzyl photosensitive linker to form a conjugate of formula II. The o-nitrobenzyl group acts as a photocleavable group, and the nitrobenzyl ring undergoes a fragmentation reaction under two-photon excitation to specifically release the borneol molecule. The structural formula of formula II is:

4. The composition of a glucose-substituted pyrazole compound and borneol according to claim 1, characterized in that: The stabilizer in the injection carrier is 2% of a freeze-drying protectant and 0.08% of an osmotic pressure regulator, with a content of 1-3%, and the cosolvent is propylene glycol; The stabilizer and propylene glycol work together to ensure the stability and biocompatibility of the preparation, and release borneol through two-photon excitation to act on targeted therapy.

5. The composition of a glucose-substituted pyrazole compound and borneol according to claim 1, characterized in that: The concentration of the supramolecular inclusion compound in the injection carrier is 0.1-5.0 mg / mL.

6. The composition of a glucose-substituted pyrazole compound and borneol according to claim 1, characterized in that: The basic auxiliary materials include: pH regulator, antioxidant, eluent, catalyst; The pH regulator is sodium hydroxide, which is used to adjust the pH value of the injection to 7.0-7.8; The antioxidant is sodium bisulfite with a concentration of 0.05-0.1%; The eluent is chloroform and methanol in a molar ratio of 8:1; The catalyst is copper sulfate (CuSO4) and sodium ascorbate in a molar ratio of 1:

2.

7. A method for preparing a composition of a glucose-substituted pyrazole compound and borneol, suitable for preparing a composition of a glucose-substituted pyrazole compound and borneol according to any one of claims 1 to 6, characterized in that: The specific steps of this method are: S100, Synthesis of the Compound of Formula I': Dissolve the compound of Formula I, 1-phenyl-3-methyl-5-OD-glucoside-pyrazole, and naphthalene imide carboxylic acid in anhydrous DMF at a molar ratio of 1:1.2, add 1.5% EDC·HCl and 1.2% NHS, and react at 25°C under nitrogen for 12 hours. After purification by adding an eluent, collect the eluate to obtain a white solid, which is the compound of Formula I'; S200, preparation of a borneol derivative of formula II: dissolving borneol and o-nitrobenzyl bromide in acetone at a molar ratio of 1:1.5, adding potassium carbonate, and reacting at reflux at 60°C for 6 hours to produce an o-nitrobenzyl-protected borneol derivative, mixing the derivative with a compound of formula I' at a molar ratio of 1:1, adding a catalyst, stirring and reacting for 12 hours, collecting and filtering the precipitated solid, which is the borneol derivative of formula II; S300, supramolecular inclusion: dissolving Formula I′ and Formula II in a propylene glycol solvent at a molar ratio of 1:1.2, stirring under ultrasound assistance for 6 hours to induce inclusion complex formation, transferring the solution to a dialysis bag, dialyzing in deionized water for 24 hours to remove unincluded substances, and freeze-drying the solution to obtain a supramolecular inclusion complex; S400, preparation of injection: adding a stabilizer to the inclusion compound solution, adjusting the pH to 7.5±0.2 with sodium hydroxide, adding physiological saline to the final volume, sterilizing and filtering through a filter membrane, and then filling into an ampoule to obtain the injection of the composition of the glucose-substituted pyrazole compound and borneol; S500, verification of light-controlled release: The injection solution was pre-frozen to -80°C and then freeze-dried to obtain a white freeze-dried powder, which was reconstituted with water for injection before use. The borneol release efficiency was verified by two-photon laser.

8. A use of a composition of a glucose-substituted pyrazole compound and borneol, suitable for the composition of a glucose-substituted pyrazole compound and borneol according to any one of claims 1 to 6, characterized in that: The injection of the composition is used in the following fields: Treatment of brain diseases: Borneol is delivered through the blood-brain barrier, and the inclusion complex is released at the site of cerebral ischemia and neuroinflammation by triggering near-infrared light, which is used to treat stroke and Alzheimer's disease; Precision tumor treatment: Two-photon excitation enables light-controlled drug release in deep tumor tissues, suitable for gliomas and breast cancer brain metastases; Real-time therapeutic monitoring: The naphthalene imide group provides fluorescent tracing function, which can monitor drug distribution and release kinetics through in vivo imaging.

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