A shikonin composition for inhibiting non-specific inflammation of the reproductive system

Abstract

This invention relates to the field of chemical pharmaceutical formulation manufacturing technology, and discloses a shikonin composition for inhibiting non-specific inflammation of the reproductive system, comprising: nanomicelles formed by encapsulating shikonin with tocopherol polyethylene glycol 1000 succinate, phenylboronic acid-modified sodium hyaluronate, and a lactic acid buffer system. The phenylboronic acid-modified sodium hyaluronate constructs a gel anchoring layer by complexing the phenylboronic acid groups with the ortho-dihydroxy structure on the action surface. This invention utilizes hydrogen peroxide to induce an oxidative deboronization reaction at the inflammatory lesion, causing an in-situ phase transition from a gel state to a sol state in the gel anchoring layer, eliminating drug diffusion resistance, and resolving the mismatch between mucosal clearance and drug release rate. The composition enhances the chemical stability of shikonin while achieving feedback regulation of drug dosage and inflammation degree, effectively repairing the mucosal barrier and maintaining local microecological balance.

Description

Technical Field

[0001] This invention relates to a shikonin composition for inhibiting non-specific inflammation of the reproductive system, belonging to the field of chemical pharmaceutical manufacturing technology. Background Technology

[0002] As a traditional and precious Chinese medicinal herb, Xinjiang Lithospermum erythrorhizon contains naphthoquinone compounds (such as shikonin and its derivatives) in its extracts, which have significant anti-inflammatory, antibacterial, and wound-healing bioactivities. In the field of reproductive system mucosal protection, Lithospermum erythrorhizon extract is often used to inhibit the growth of harmful bacteria and accelerate the healing of damaged tissues. However, natural shikonin has extremely strong hydrophobicity and chemical instability, and the direct use of Lithospermum erythrorhizon extract often faces technical bottlenecks such as low bioavailability and poor mucosal permeability. Currently, the local treatment of non-specific inflammation of the reproductive system uses shikonin as the core drug source. Usually, cellulose derivatives or synthetic polymer materials are used to construct a gel matrix to carry hydrophobic active ingredients and form a physical reservoir on the mucosal surface. The cavity environment is in a dynamic cycle of body fluid flushing and physiological secretion. The residence time of the formulation depends on the rheological properties of the matrix and the interfacial adhesion strength. In order to counteract the mechanical stripping effect of physiological fluids, the formulation design establishes a highly dense polymer topological network, and prolongs the drug action period by improving the energy modulus and mechanical strength of the matrix.

[0003] This logic based on static physical locking creates a constraint between formulation retention capacity and drug diffusion kinetics. While the dense polymer network provides erosion resistance, the compression of intermolecular spaces creates mass transfer resistance, making it difficult for shikonin molecules encapsulated within the matrix to overcome cohesive forces and migrate to the lesion tissue. The actual release rate is lower than the effective anti-inflammatory concentration. To compensate for this release resistance, the initial drug loading is increased, resulting in overloaded chemical stimulation in non-lesion areas and disrupting the lactobacillus-dominated microecological balance, leading to a cycle of recurrent inflammation. Besides the limitations of the physical carrier morphology, the intelligent release logic of the formulation for complex pathological environments also has shortcomings. For example, the Chinese patent with authorization announcement number CN112237635B... The patent discloses an active targeted antitumor nanomicelle, its preparation method, and its application. By modifying the surface of the nanocomposite with specific antibodies, active drug targeting is achieved through antigen-antibody biorecognition. This type of technology relies on the overexpression of specific diseased cell surface antigens, which requires the existence of clear and stable biomolecular targets. Under non-specific inflammatory conditions in the reproductive system, lesions are characterized by the widespread presence of oxidative stress signals. The aforementioned scheme cannot identify non-specific inflammatory chemical signals, and the static antibody recognition mechanism is difficult to adjust the dosage in response to the dynamic evolution of the degree of inflammation. This scheme lacks the ability to simultaneously intervene in mucosal barrier damage and microecological pH imbalance, and there is a logical mismatch when treating mucosal inflammation with high flushing and non-specific characteristics.

[0004] Therefore, how to achieve precise in-situ drug delivery to lesions through multi-mechanism coupling is the technical problem to be solved by this invention. Summary of the Invention

[0005] To address the problems mentioned in the background art, the technical solution of the present invention is as follows: a shikonin composition for inhibiting non-specific inflammation of the reproductive system, the composition comprising:

[0006] Shikonin micelle particles, phenylboronic acid-modified sodium hyaluronate, and lactate-sodium lactate buffer; based on the total mass of the composition (100%), the mass percentage of shikonin micelle particles is 5% to 12%, the mass percentage of phenylboronic acid-modified sodium hyaluronate is 1.5% to 3.5%, and the balance is lactate-sodium lactate buffer.

[0007] Shikonin micelles are nanoparticles assembled from tocopherol polyethylene glycol 1000 succinate and shikonin, wherein the hydrophilic-lipophilic balance value of tocopherol polyethylene glycol 1000 succinate is 13 to 13.5, and the mass ratio of tocopherol polyethylene glycol 1000 succinate to shikonin is 10:1 to 15:1.

[0008] Phenylated boric acid modified sodium hyaluronate is sodium hyaluronate grafted with phenylboronic acid groups. The weight-average molecular weight of the sodium hyaluronate is 800 kDa to 1200 kDa, and the grafting degree of phenylboronic acid groups on the sodium hyaluronate backbone is 18 mol% to 22 mol%.

[0009] The composition The value is 3.8 to 4.2; wherein, the phenylboronic acid group has the chemical activity to form a borate ester complexation reaction with the ortho-dihydroxy structure to form a gel anchoring layer at the interface of the composition; in an environment with a hydrogen peroxide concentration of 0.1 mmol / L to 1.0 mmol / L, the phenylboronic acid group undergoes an oxidative deboronization reaction and is converted into phenolic hydroxyl groups, causing the gel anchoring layer to undergo a phase transition from a gel state to a sol state to release shikonin.

[0010] Preferably, the average particle size of the shikonin micelles is 35 nm to 60 nm; wherein, the hydrophobic core formed by tocopherol polyethylene glycol 1000 succinate physically encapsulates the shikonin, thereby spatially isolating the naphthoquinone core of the shikonin from the aqueous environment in the lactate-sodium lactate buffer solution.

[0011] Preferably, the shear yield stress of the phenylboronic acid-modified sodium hyaluronate in the composition system is greater than 50 Pa; when hydrogen peroxide permeates into the gel anchoring layer, the oxidative deboronization reaction causes the crosslinking points in the gel anchoring layer to break.

[0012] Preferably, the lactate-sodium lactate buffer solution has a lactate concentration of 0.1 mol / L to 0.3 mol / L, used for initial preparation of the composition system. The value remained between 3.8 and 4.2, and after the crosslinking point broke, it maintained the hydrogen ion concentration in the local microenvironment.

[0013] Preferably, the shikonin micelle particles include an active component that inhibits cell membrane efflux pumps; after the cross-linking points break, the shikonin micelle particles interact with the cell membrane via their tocopherol polyethylene glycol 1000 succinate component. - Glycoprotein binding sites are physically occupied.

[0014] Preferably, the phenylboronic acid group is a chemical structure formed by an amidation reaction between aminophenylboronic acid and the carboxyl group on the main chain of sodium hyaluronate; the molar ratio of the condensing agent to the carboxyl group in the amidation reaction is 1.2:1 to 1.5:1.

[0015] Preferably, the composition has a dynamic viscosity of 1500 mPa·s to 3500 mPa·s at 25°C; the phenylboronic acid-modified sodium hyaluronate forms physical crosslinking points composed of phenylboronic acid groups in the composition system through the coupling of weight-average molecular weight and grafting degree.

[0016] Preferably, the method for preparing shikonin micelle particles includes: step S91: dissolving shikonin and tocopherol polyethylene glycol 1000 succinate in anhydrous ethanol, and removing the anhydrous ethanol by rotary evaporation under vacuum at 40°C to 45°C to form a film; step S92: adding lactate-sodium lactate buffer to the film formed in step S91, and hydrating it at 55°C to 60°C.

[0017] Preferably, the dosage form of the composition is a gel or suppository; the composition is used to repair the damaged physical barrier by physically covering the action surface with sodium hyaluronate in response to changes in hydrogen peroxide concentration.

[0018] Compared with the prior art, the beneficial effects of the present invention are:

[0019] 1. In the shikonin composition for inhibiting non-specific inflammation of the reproductive system, the responsive anchoring matrix resolves the spatiotemporal mismatch between mucosal retention and drug release during reproductive system administration by establishing a dynamic responsive network of phenylboronic acid grafted with hyaluronic acid. The phenylboronic acid groups form reversible borate ester bonds with mucosal glycoproteins, providing chemical anchoring force to resist physiological fluid flushing and replacing the physical thickening of traditional high-concentration polymers. When hydrogen peroxide in the inflammatory lesion area penetrates into the matrix, the oxidative deboronization reaction induces the breakage of polymer crosslinking points, causing the formulation to undergo a phase transition from gel to sol in situ at the lesion, eliminating the spatial resistance to drug diffusion and achieving feedback regulation of drug dosage and pathological signal intensity.

[0020] 2. The synergistic effect of solubilizing micronuclei and dynamic response cross-linking networks establishes a pathway for ensuring the chemical stability and enhancing the penetration of shikonin in complex physiological environments. The hydrophobic core composed of tocopherol polyethylene glycol 1000 succinate isolates shikonin from the external aqueous environment, blocking the oxidative degradation of naphthoquinone nucleus in the body fluid environment. When pathological signals trigger the topological collapse of the anchoring matrix, the freed micronuclei utilize their inhibitory properties on the cell membrane efflux pump to guide shikonin to overcome the transmission resistance of mucosal basal cells and increase the effective drug dosage in the deep lesion.

[0021] 3. The interaction logic between pH maintenance solution and responsive anchoring matrix constructs an adaptive physical blocking loop at the edge of the lesion. Utilizing the acid-base dependence of the binding constant of phenylboronic acid and diol, the formulation releases the drug through oxidative degradation in the core area of ​​the lesion, while maintaining a high cross-linking density in the peripheral area where the pH value rises due to the influence of pathogenic bacteria. This spatially differentiated response logic blocks the lateral spread of non-specific inflammation while maintaining the stability of the lactobacillus microecology.

[0022] 4. This invention utilizes the synergistic effect of Xinjiang Lithospermum extract with intelligent responsive nanomicelles and dynamic gel networks to achieve precise matching between the active components of Lithospermum extract and pathological environmental signals. Based on leveraging the antibacterial and anti-inflammatory effects of Lithospermum, it effectively enhances the biological performance of the formulation in repairing the mucosal barrier through physical coverage of the damaged mucosal surface and dosage feedback regulation, providing a systematic technical path for non-specific inflammation of the reproductive system based on the integration of authentic medicinal materials and modern drug delivery technology. Attached Figure Description

[0023] Figure 1 This is a diagram illustrating the mechanism of hydrogen peroxide-triggered gel phase transition and precise drug release of shikonin in this invention.

[0024] Figure 2 This is a technical logic diagram of the multi-effect synergistic inhibition of reproductive system inflammation according to the present invention.

[0025] The objectives, features, and advantages of this invention will be further explained in conjunction with the embodiments and with reference to the accompanying drawings. Detailed Implementation

[0026] The technical solutions of the embodiments of this application will be clearly described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of this application, not all embodiments. All other embodiments obtained by those skilled in the art based on the embodiments of this application are within the scope of protection of this application.

[0027] A shikonin composition for inhibiting non-specific inflammation of the reproductive system, the composition comprising:

[0028] Shikonin micelle particles, phenylboronic acid-modified sodium hyaluronate, and lactate-sodium lactate buffer; based on the total mass of the composition (100%), the mass percentage of shikonin micelle particles is 5% to 12%, the mass percentage of phenylboronic acid-modified sodium hyaluronate is 1.5% to 3.5%, and the balance is lactate-sodium lactate buffer.

[0029] Shikonin micelles are nanoparticles assembled from tocopherol polyethylene glycol 1000 succinate and shikonin, wherein the hydrophilic-lipophilic balance value of tocopherol polyethylene glycol 1000 succinate is 13 to 13.5, and the mass ratio of tocopherol polyethylene glycol 1000 succinate to shikonin is 10:1 to 15:1.

[0030] Phenylated boric acid modified sodium hyaluronate is sodium hyaluronate grafted with phenylboronic acid groups. The weight-average molecular weight of the sodium hyaluronate is 800 kDa to 1200 kDa, and the grafting degree of phenylboronic acid groups on the sodium hyaluronate backbone is 18 mol% to 22 mol%.

[0031] The composition The value is 3.8 to 4.2; wherein, the phenylboronic acid group has the chemical activity to form a borate ester complexation reaction with the ortho-dihydroxy structure to form a gel anchoring layer at the interface of the composition; in an environment with a hydrogen peroxide concentration of 0.1 mmol / L to 1.0 mmol / L, the phenylboronic acid group undergoes an oxidative deboronization reaction and is converted into phenolic hydroxyl groups, causing the gel anchoring layer to undergo a phase transition from a gel state to a sol state to release shikonin.

[0032] Preferably, the average particle size of the shikonin micelles is 35 nm to 60 nm; wherein, the hydrophobic core formed by tocopherol polyethylene glycol 1000 succinate physically encapsulates the shikonin, thereby spatially isolating the naphthoquinone core of the shikonin from the aqueous environment in the lactate-sodium lactate buffer solution.

[0033] Preferably, the shear yield stress of the phenylboronic acid-modified sodium hyaluronate in the composition system is greater than 50 Pa; when hydrogen peroxide permeates into the gel anchoring layer, the oxidative deboronization reaction causes the crosslinking points in the gel anchoring layer to break.

[0034] Preferably, the lactate-sodium lactate buffer solution has a lactate concentration of 0.1 mol / L to 0.3 mol / L, used for initial preparation of the composition system. The value remained between 3.8 and 4.2, and after the crosslinking point broke, it maintained the hydrogen ion concentration in the local microenvironment.

[0035] Preferably, the shikonin micelle particles include an active component that inhibits cell membrane efflux pumps; after the cross-linking points break, the shikonin micelle particles interact with the cell membrane via their tocopherol polyethylene glycol 1000 succinate component. - Glycoprotein binding sites are physically occupied.

[0036] Preferably, the phenylboronic acid group is a chemical structure formed by an amidation reaction between aminophenylboronic acid and the carboxyl group on the main chain of sodium hyaluronate; the molar ratio of the condensing agent to the carboxyl group in the amidation reaction is 1.2:1 to 1.5:1.

[0037] Preferably, the grafting degree of the phenylboronic acid groups Satisfy the following formula: ,in, This refers to the amount of phenylboronic acid groups grafted onto the sodium hyaluronate backbone. The amount of substance of repeating units in sodium hyaluronate; grafting degree. The values ​​range from 18% to 22%.

[0038] Preferably, the composition has a dynamic viscosity of 1500 mPa·s to 3500 mPa·s at 25°C; the phenylboronic acid-modified sodium hyaluronate forms physical crosslinking points composed of phenylboronic acid groups in the composition system through the coupling of weight-average molecular weight and grafting degree.

[0039] Preferably, the method for preparing shikonin micelle particles includes: step S91: dissolving shikonin and tocopherol polyethylene glycol 1000 succinate in anhydrous ethanol, and removing the anhydrous ethanol by rotary evaporation under vacuum at 40°C to 45°C to form a film; step S92: adding lactate-sodium lactate buffer to the film formed in step S91, and hydrating it at 55°C to 60°C.

[0040] Preferably, the dosage form of the composition is a gel or suppository; the composition is used to repair the damaged physical barrier by physically covering the action surface with sodium hyaluronate in response to changes in hydrogen peroxide concentration.

[0041] Example 1: When the system faces dynamic fluid flushing of the reproductive system mucosa, the local microenvironment exhibits a weakly acidic state with a pH of 3.8 to 4.2 corresponding to the basement hydrogen ion concentration; non-specific inflammatory lesions show hydrogen peroxide overexpression at concentrations of 0.1 mmol / L to 1.0 mmol / L; conventional formulations construct a dense polymer topological network to provide resistance to scouring shear stress. This topological network increases adhesion and retention capacity while compressing the internal free volume, leading to a decrease in the diffusion coefficient of drug molecules within the matrix and causing mass transfer hindrance between adhesion retention and release pathways. A shikonin composition for inhibiting non-specific inflammation of the reproductive system includes shikonin micelle particles, phenylboronic acid-modified sodium hyaluronate, and lactate-sodium lactate buffer. The composition reconstructs the mass transfer process through a specific component ratio and a response phase transition mechanism. Sodium hyaluronate with a weight-average molecular weight of 800 kDa to 1200 kDa is selected as the main chain, and phenylboronic acid-modified sodium hyaluronate is constructed by grafting aminophenylboronic acid via an amidation reaction. The molar ratio of the condensing agent to the carboxyl groups on the sodium hyaluronate main chain is controlled to be 1.2:1 to 1.5:1 during the amidation reaction, so that the grafting degree of the phenylboronic acid groups follows a mathematical relationship. The calculation formula is as follows: ;in, This refers to the amount of phenylboronic acid groups grafted onto the sodium hyaluronate backbone. The amount of substance of the repeating unit in sodium hyaluronate, and the grafting degree The value is limited to 18% to 22%; tocopherol polyethylene glycol 1000 succinate with a hydrophilic-lipophilic balance of 13 to 13.5 and shikonin are selected and assembled at a mass ratio of 10:1 to 15:1 to form shikonin micelles with an average particle size of 35nm to 60nm; in the preferred embodiment of the present invention, shikonin is preferably obtained from high-quality Xinjiang shikonin extract, which is obtained by ultrasonic-assisted extraction and recrystallization purification to ensure its high purity and bioactivity in the subsequent micelle assembly process. The core component of this shikonin extract not only has significant anti-inflammatory and broad-spectrum antibacterial abilities, effectively inhibiting pathogenic bacteria such as Staphylococcus aureus, Escherichia coli, and Candida albicans, but also works synergistically with sodium hyaluronate in the complex inflammatory environment of the reproductive system to repair mucosa and promote epithelial tissue remodeling; lactate-sodium lactate buffer is prepared with lactate at a concentration of 0.1mol / L to 0.3mol / L; on this basis, lactate-sodium lactate buffer is prepared. Sodium hyaluronate buffer is added as the balance to a mixture containing 5% to 12% by mass of shikonin micelle particles and 1.5% to 3.5% by mass of phenylboronic acid-modified sodium hyaluronate. The initial pH of the composition is maintained at 3.8 to 4.2. Under this weakly acidic constraint, the phenylboronic acid groups maintain an electrically neutral configuration. The shikonin micelle particles are uniformly dispersed and physically trapped in the interstices of the polymer network, forming a pristine gel with a dynamic viscosity of 1500 mPa·s to 3500 mPa·s at 25°C. In this system, the active molecules derived from Xinjiang shikonin are locked in the nanospace. Through physical and chemical dual anchoring, the problem of easy loss of traditional shikonin extracts on the mucosal surface is solved. This design allows the formulation to exhibit the anti-inflammatory efficacy of shikonin while significantly improving the ability to repair damaged mucosa by utilizing the biocompatibility of sodium hyaluronate, achieving a three-in-one therapeutic effect of antibacterial, anti-inflammatory, and repair.

[0042] After the composition is applied to the mucosal surface, the phenylboronic acid groups on the side chains of sodium hyaluronate modified with phenylboronic acid exhibit chemical activity, recognizing the exposed cis-diol structure of the mucosa glycoprotein molecules. The two undergo dehydration condensation to form a borate ester covalent bond, constructing a gel anchoring layer at the composition interface with a shear yield stress greater than 50 Pa. The hydrophobic core formed by tocopherol polyethylene glycol 1000 succinate physically encapsulates shikonin, spatially isolating the naphthoquinone nucleus of shikonin from the aqueous environment in the lactate-sodium lactate buffer, blocking the oxidative degradation pathway. When the gel anchoring layer covers the area... In the presence of hydrogen peroxide molecules at concentrations ranging from 0.1 mmol / L to 1.0 mmol / L, hydrogen peroxide permeates into the gel anchoring layer and triggers an oxidative deboronization reaction, oxidizing and trunculating the carbon-boron bonds and converting them into phenolic hydroxyl groups. The breaking of these chemical bonds leads to the rupture of physical cross-linking points composed of phenylboronic acid groups, causing the in-situ gel anchoring layer to undergo a phase transition from a gel state to a sol state. This phase transition process essentially manifests as a unidirectional, progressive physical erosion mechanism advancing from the mucosal contact interface into the matrix interior. The trace amounts of hydrogen peroxide released from the mucosal substrate come into contact with the bottommost phenylboronic acid ester bonds. This triggers rapid oxidative cleavage, causing the local polymeric topological network at the micro-region interface to untangle first, generating a sol-gel-like transitional aqueous phase thin layer with a thickness on the order of micrometers in situ. At this point, the outer gel matrix, which has not yet been touched by the chemical signal of hydrogen peroxide, still maintains an intact cross-linked network and a shear yield stress greater than 50 Pa, continuously resisting the outward scouring shear force of body fluid in the overall environment; while the free and detached nano-shikonin micelles in the inner transition layer are unaffected by the external scouring flow field, and thus diffuse and penetrate into the deep inflammatory base layer by virtue of the concentration gradient, thereby affecting the entire... A physically self-consistent kinetic equilibrium loop is established between the high shear fluid stasis and the targeted and precise release from superficial lesions. The shear yield stress of the composition system is determined by standardized rheological testing procedures. A rotational rheometer with a 40 mm cone-plate clamp is used, and the temperature control module is turned on to maintain a constant test platform temperature of 37°C. An appropriate amount of composition sample is placed in the clamp gap and allowed to stand for 3 minutes to equilibrate. The instrument is set to the working stress scanning mode, and the output shear stress is continuously scanned from 0.1 Pa to 200 Pa in a logarithmic increment. The test platform simultaneously collects and records the storage modulus. and loss modulus The energy storage modulus was extracted from the response curve of shear stress variation. Curve and loss modulus The shear stress coordinate value corresponding to the intersection and coincidence point of the curves is calibrated as the shear yield stress of the system. As the cross-linking point breaks and the topological structure collapses, the shikonin micelle particles that are freed from the steric hindrance are released into the body fluid and release shikonin. The free shikonin micelle particles utilize the tocopherol polyethylene glycol 1000 succinate component to physically occupy the P-glycoprotein binding sites on the basal cell membrane, thereby eliminating the steric hindrance of drug diffusion.

[0043] Simultaneously, the lactate-sodium lactate buffer solution maintains the hydrogen ion concentration in the local microenvironment after the cross-linking points break, keeping the surrounding non-lesion areas unaffected by pathological signals in a weakly acidic state and maintaining the cross-linking density of the physical blocking rings. The composition responds to changes in hydrogen peroxide concentration by releasing the active component in situ, and the surrounding sodium hyaluronate physically covers the action surface, repairing the damaged physical barrier. During the release phase, the shikonin micelles detach from the gel topological network and rely on the tocopherol polyethylene glycol 1000 succinate component arranged in the outer layer to exert transmembrane transport effects. The tocopherol polyethylene glycol 1000 succinate component, as an active component inhibiting cell membrane efflux pumps, directly occupies the P-glycoprotein binding sites on the cell membrane surface after contacting basal cells. Specifically, the nanomicelles dissociate... Subsequently, the hydrophilic polyethylene glycol segment of the tocopherol polyethylene glycol 1000 succinate amphiphilic molecule exhibits a highly flexible coiled conformation in an aqueous physiological environment. This allows it to form a multi-layered, tight hydrogen bond network with amino acid residues surrounding the ATP-binding site in the transmembrane domain of the P-glycoprotein, which is responsible for capturing substrates. Simultaneously, the rigid hydrophobic tail of the tocopherol is spontaneously driven by hydrophobic forces to deeply embed and anchor itself within the core substrate-binding lumen of the efflux pump protein. This highly matched and fused hydrophilic and hydrophobic dual domains in the surface spatial conformation directly seals the drug efflux channel of the P-glycoprotein at the molecular topological level, constructing a rigid steric barrier that blocks the energy-dependent pumping pathway of the P-glycoprotein to maintain the concentration of shikonin within the target cell.

[0044] Example 2: In verifying the pathological condition of dynamic fluid flushing on the surface of the reproductive system mucosa intertwined with local reactive oxygen species overexpression, fluid shear force testing and drug release kinetic characterization constitute the test path for evaluating the mass transfer properties of the composition. A microfluidic mucosal permeation and release test platform was used to construct the test environment. The platform included a thermostatic supply pump, a simulated mucosal substrate, and a multi-channel collection end. The output temperature of the thermostatic supply pump was set to 37°C. The basic perfusion fluid was set to a lactate-sodium lactate buffer solution with a pH value of 4.0 corresponding to the hydrogen ion concentration. The setting of fluid flow rate parameters needed to balance the engineering contradiction between simulating physiological clearance rate and maintaining the stability of the observation window. When the perfusion fluid flow rate was too low, effective shear stress could not be generated, and when the flow rate was too high, the drug detection concentration would be lower than the instrument's lower limit of quantitation. Based on the physiological indicators of mucosal secretion, the basic flow rate was set to 2.5 mL / h. At the same time, a random flow rate fluctuation signal with an amplitude of 5% was superimposed in the control program to introduce fluid disturbance noise caused by physiological muscle group contraction.

[0045] Multiple sets of composite samples were prepared to form a multi-dimensional control system. A phenylboronic acid-modified sodium hyaluronate with a grafting degree of 20% was mixed with tocopherol polyethylene glycol 1000 succinate and shikonin micelle particles at a mass ratio of 12:1 to form the sample group of this invention with complete technical features. A control group exceeding the range was constructed with a corresponding component having a grafting degree of 10%. A control group exceeding the range was constructed with a corresponding component having a grafting degree of 30%. A partially missing control group was constructed by replacing the core component with sodium hyaluronate without grafted phenylboronic acid groups. A conventional control group was constructed by loading an equivalent dose of shikonin micelle particles onto poloxamer thermosensitive gel. Each sample group was then coated with... The sample was applied to the surface of a simulated mucosal substrate; a constant-temperature supply pump was started to inject the basic perfusion fluid; dynamic flushing was applied for 120 minutes; experimental data showed that the mass retention rate of the partially missing control group decreased to 28.5% after 60 minutes of flushing, while the final mass retention rate of the out-of-range control group was 45.2%; this data indicates that the lack of sufficient phenylboronic acid ester complex bonds makes the system difficult to resist fluid shear noise; the mass retention rate of the sample group of this invention was maintained at 89.4%, the mass retention rate of the conventional control group was 88.7%, and the mass retention rate of the out-of-range control group was 91.3%. The experimental data proved that phenylboronic acid groups with a grafting degree greater than 18% provide a stable gel interface anchoring effect.

[0046] Maintaining a dynamic flushing state, hydrogen peroxide was injected into the basal perfusion fluid to a concentration of 0.5 mmol / L to provide oxidative stress signals to the inflammatory lesions. The cumulative release of shikonin was recorded over the following 180 minutes. After the introduction of hydrogen peroxide, the cumulative release in the sample group of this invention showed a stepwise increase, reaching 76.8% at 120 minutes. The conventional control sample group, constrained by the three-dimensional space of the dense network, showed a linear and slow increase in cumulative release, with the endpoint value maintained at 18.5%. The cumulative release in the out-of-range control group II showed a non-linear slowdown and flattening after reaching 42.4%. This inflection point indicates that excessively high grafting degree produces an overly dense cross-linked network. Although some carbon-boron bonds undergo oxidative deboration, the remaining physical crosslinking points maintain the original gel state, hindering phase transition. The out-of-range control group had a detection rate of less than 15% due to excessive erosion in the early stage. The above dynamic measurement data confirms that the grafting degree is defined within a numerical window of 18% to 22%, which constitutes a technically balanced working range. The composition relies on the component characteristics within this range to maintain high-strength interfacial anchorage in a physical erosion environment with fluid disturbance noise, and triggers a phase transition mechanism that promotes structural disintegration when a specific concentration of hydrogen peroxide chemical signal is detected. The scheme eliminates the spatial resistance of drug diffusion by conditionally breaking chemical bonds, overcoming the mutual exclusion between physical adhesion and drug release pathway.

[0047] Example 3: When preparing a shikonin composition to inhibit non-specific inflammation of the reproductive system, maintaining the batch stability of the grafting degree of phenylboronic acid-modified sodium hyaluronate and the uniformity of the particle size of shikonin micelles affects the phase transition response rate of the system; the conventional synthetic route does not control the lifetime of the intermediate state of the condensation reaction, and the steric hindrance of the polymer side chain increases, hindering the subsequent borate complexation reaction; a morpholine ethanesulfonic acid buffer with a concentration of 0.1 mol / L and a pH of 5.5 is prepared; sodium hyaluronate with a weight-average molecular weight of 800 kDa to 1200 kDa is dissolved in the morpholine ethanesulfonic acid buffer to form a matrix solution with a mass concentration of 1%; a carbodiimide condensing agent and hydroxyl group are added to the matrix solution. Succinimide; the molar ratio of the condensing agent to the carboxyl groups on the sodium hyaluronate backbone was controlled at 1.2:1; the reaction was activated by magnetic stirring for 2 hours at 25°C in the dark to generate a succinimide ester intermediate; aminophenylboronic acid was added, and the reaction system was stirred at 25°C for 24 hours to initiate the amidation reaction; at the end of the reaction, unreacted small molecule impurities were removed by dialyzing in deionized water for 72 hours using a dialysis bag with a molecular weight cutoff of 14 kDa, and phenylboronic acid-modified sodium hyaluronate solid powder was obtained by freeze-drying; shikonin has hydrophobicity and chemical instability, and the aggregation phenomenon caused by direct dispersion changes the local mass transfer distribution of the composition; in order to overcome this problem, this embodiment uses a new Using Lithospermum erythrorhizon extract as the active source, the naturally occurring lipid components and synthetic tocopherol polyethylene glycol 1000 succinate produce a synergistic effect. The Lithospermum erythrorhizon extract, precisely obtained through an ultrasound-assisted process, maintains its antibacterial activity while allowing its molecular framework to be better embedded in the hydrophobic core of nanomicelles. This significantly enhances the physical adhesion of the formulation to the mucosa repairing the lesion surface, while maintaining the anti-inflammatory effect of Lithospermum erythrorhizon. Tocopherol polyethylene glycol 1000 succinate, with a hydrophilic-lipophilic balance value of 13 to 13.5, is selected as an amphiphilic carrier. It is dissolved in anhydrous ethanol with shikonin at a mass ratio of 15:1 to form a homogeneous solution. The homogeneous solution is then placed in a rotating... In an evaporator, anhydrous ethanol was removed by rotary evaporation under a water bath temperature of 40℃ and a vacuum of 0.08MPa, causing the solute to deposit on the inner wall of the flask to form a solid film. The solid film was then hydrated and eluted using a 0.2mol / L lactate-sodium lactate buffer solution. The hydration temperature was controlled at 37℃, and ultrasonic oscillation with a power of 100W was applied for 30 minutes. The hydrophobic ends of tocopherol polyethylene glycol 1000 succinate were driven to encapsulate shikonin molecules inward while the hydrophilic ends extended outward, self-assembling to form shikonin micelle particles with an average particle size of 45nm. The hydrated solution was filtered using a microporous membrane with a pore size of 0.22μm to remove free drug and aggregates, obtaining a shikonin micelle particle dispersion.

[0048] The composition is formed by the spatial physical interlocking of macromolecular networks and nanomicelles. Phenylated boric acid-modified sodium hyaluronate powder is added to a dispersion of shikonin micelle particles, and the mixture is stirred at 300 rpm at room temperature for 4 hours using a mechanical stirrer to allow the polymer segments to swell and expand in the buffer system. Lactic acid or sodium lactate solution is added dropwise to the mixture to adjust the pH to 4.0. The electroneutrally neutral configuration of the phenylboronic acid groups is maintained under weak acid constraints to inhibit cross-linking and aggregation. The mass of the added components is controlled to ensure that the mass percentage of shikonin micelle particles in the composition reaches 8%, and the mass percentage of phenylboronic acid-modified sodium hyaluronate reaches [missing value]. 2.5%; the preparation procedure controls the grafting degree of the polymer grafting reaction and the particle size during the drug self-assembly process; the resulting composition exhibits a primary gel structure in which micelle particles are distributed within a weakly acidic polymer network, undergoing an oxidative deboronization phase transition when exposed to hydrogen peroxide stimulation from the lesion. To address the limitation of terminal sterilization filtration technology due to the system's dynamic viscosity being between 1500 mPa·s and 3500 mPa·s, a combined aseptic dispensing and assembly process is used to shape the composition. A dispersion of shikonin micelle particles with an average particle size of 35 nm to 60 nm and a lactate-sodium lactate buffer solution are respectively passed through a 0.22 μm polyethersulfone filter membrane for sterilization filtration. After the synthesis of phenylboronic acid-modified sodium hyaluronate solid powder, an electron beam dose of 10 kGy to 15 kGy was applied to sterilize the monomer and control the degradation rate of the polymer backbone. During this energy input and conversion process, the high-energy penetrating electron beam directly bombarded and targeted the β-1,4-glycosidic bond structure in the hyaluronic acid backbone. Based on the dose-effect mathematical correlation curve of excited chain scission of polymers obtained in the previous stage, the controlled low-dose radiation window of 10 kGy to 15 kGy can accurately excite and generate only a limited number of active free radicals, so that the polymer backbone only undergoes sporadic breakage and ultimately the weight-average molecular weight of the material is 5. With a controlled buoyancy of 8% to 10%, this precise micro-degradation at this level successfully delivers sufficient energy to destroy the potential microbial nucleic acid structure load within the powder, while avoiding the physicochemical risks of large-scale depolymerization and fragmentation of the polymer molecular skeleton. This solidly ensures that the polymer powder can still maintain a robust mechanical strength baseline during the subsequent hydration and reconstruction stage. In a sterile operating area that meets the Class A cleanliness standard, the irradiated sterilized polymer powder is put into a sterile homogenizing reactor, and the sterile filtered liquid phase component is injected using a sterile pumping system. The high-shear mixing device is started and stirred and hydrated at a low temperature of 10°C to 15°C to complete the aseptic preparation assembly.

[0049] Example 4: When facing fluctuations in the molecular weight distribution of sodium hyaluronate raw materials and batch-to-batch differences in the activity of condensing agents, the grafting degree synthesis route tends to deviate from the set range. The reaction kinetic parameters of the sodium hyaluronate backbone and aminophenylboronic acid were quantified. A pre-synthesized phenylboronic acid-modified sodium hyaluronate intermediate sample was extracted and dissolved in a heavy water solvent containing deuterated hydrochloric acid to form a 5 mg / mL NMR test solution. Proton NMR spectra were acquired using a pulsed Fourier transform NMR spectrometer at a working frequency of 500 MHz. The integrated area of ​​the benzene ring proton absorption peak with a chemical shift of 7.4 to 7.8 was extracted from the spectral data as the first characteristic parameter, and the integrated area of ​​the sodium acetaminomethyl proton absorption peak with a chemical shift of 1.9 was extracted as the second characteristic parameter. The quotient of the first characteristic parameter to the fourth and the quotient of the second characteristic parameter to the third were calculated, and the ratio of these two quotients was extracted as the amount of phenylboronic acid groups grafted onto the sodium hyaluronate backbone. The amount of substance of repeating units in sodium hyaluronate Based on the proportion data, calculate the initial grafting degree of the intermediate samples. .

[0050] Based on the initial grafting degree data, the formulation fine-tuning and compensation operation is initiated; the measured initial grafting degree is compared with the quantitative range of 18% to 22%; if the initial grafting degree is lower than 18%, an equivalent amount of aminophenylboronic acid and carbodiimide condensing agent is added to the main reactor, and the stirring time is extended by 2 hours; if the initial grafting degree is higher than 22%, a set mass ratio of free sodium hyaluronate solution is added to reorganize the polymer network space; this reorganization mechanism does not refer to forcibly driving the reverse bond breaking and stripping reaction on the irreversibly solidified covalent bond nodes, but rather fully utilizes the spontaneous swelling, extension, and physical entanglement characteristics of long polymer chains in the liquid phase environment. The newly added free unmodified sodium hyaluronate macromolecular chain segments spatially interpenetrate and deeply penetrate the original over-grafted polymer chains through dense intermolecular hydrogen bonding. Mixing effectively dilutes the relative molar spatial distribution density of the highly chemically active phenylboronic acid side groups at the overall fluid volume level. This three-dimensional dilution effect in the physical dimension allows the average physical spacing between the cross-linking points of the borate ester bonds to be proportionally stretched and amplified when the system finally cross-links and solidifies on the contact mucosa surface. Thus, without destroying the physical structure of the main chain, the apparent average grafting degree of the entire matrix network is effectively reduced. After parameter compensation, proton nuclear magnetic resonance spectra are collected again and the integrated area is compared until the measured grafting degree falls within the target range. Spectral analysis and mass compensation suppress the interference of raw material property fluctuations on the steric hindrance of polymer side chains, enabling the composition to maintain a consistent borate ester complexation reaction rate and the physical state of hydrogen peroxide-triggered phase transition response between batches.

[0051] Example 5: When the preparation process faces fluctuations in the hydrophilic and lipophilic properties of the carrier material and individual differences in the buffering capacity of the lesion exudate, the particle size distribution of the shikonin micelles and the maintenance capacity of the buffer system fluctuate. An online dynamic light scattering feedback loop is used to calibrate the ultrasonic parameters of the hydration self-assembly. During the initial stage of hydration ultrasonic oscillation, the mixture is continuously extracted and guided into the sample cell of the dynamic light scattering instrument. A laser light source is used to irradiate the sample cell, and the autocorrelation function of the scattered light intensity is collected. The decay rate of the autocorrelation function is analyzed using the cumulative method, and the hydrodynamic radius is extracted as the average particle size of the micelles. The warning particle size threshold is set to 60 nm and the polydispersity index threshold is set to 0.2; a comparison model is preset in the controller, and the average particle size of three consecutive sampling periods is used as the warning threshold. When the wavelength is greater than 60 nm and the polydispersity index is less than 0.2, the controller outputs a duty cycle adjustment signal to increase the output power of the ultrasonic generator and increase the physical shear strength of the ultrasonic cavitation effect. When the polydispersity index is detected to be greater than or equal to 0.2, the controller outputs an abnormal electrical signal to trigger the shut-off valve to close and isolate the corresponding batch of materials. This feedback loop corrects the ultrasonic energy input based on the change in light scattering signal to suppress the interference of raw material batch differences on the drug self-assembly process.

[0052] Before preparing the lactate-sodium lactate buffer solution, a baseline calibration method for acid-base buffer capacity was used. A sample of exudate from the reproductive system mucosa simulating a pathological condition was extracted and placed in a constant-temperature titration vessel. A sodium hydroxide standard solution of known molar concentration was added dropwise to the exudate sample at a uniform rate using an automatic potentiometric titrator. The pH response curve of the exudate sample as a function of the added volume was recorded simultaneously. The volume of sodium hydroxide standard solution consumed when the pH increased by one unit from the initial state was extracted. The product of the consumed volume and the standard concentration was calculated and divided by the initial volume of the exudate sample to obtain the exudate buffer capacity. Set the buffer maintenance factor The value is 3; the exudate buffer capacity is... With buffer maintenance coefficient The product of the values ​​is the set value of the target buffer capacity. Based on this set value and the weak acid dissociation constant of lactic acid, the molar ratio and absolute concentration of lactic acid and sodium lactate are calculated. The lactic acid raw material and sodium lactate raw material are weighed according to the calculated molar ratio to prepare the buffer solution. This calibration method calculates the buffer ratio based on the acid-base titration response data so that the prepared buffer solution maintains the pH value of the microenvironment in the range of 3.8 to 4.2 during physiological flushing.

[0053] It will be apparent to those skilled in the art that the present invention is not limited to the details of the exemplary embodiments described above, and that the present invention can be implemented in other specific forms without departing from the spirit or essential characteristics of the present invention.

[0054] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention and are not intended to limit it. Although the present invention has been described in detail with reference to preferred embodiments, those skilled in the art should understand that modifications or equivalent substitutions can be made to the technical solutions of the present invention without departing from the spirit and scope of the technical solutions of the present invention.

Claims

1. A shikonin composition for inhibiting non-specific inflammation of the reproductive system, characterized in that the composition... include: Shikonin micelle particles, phenylboronic acid-modified sodium hyaluronate, and lactate-sodium lactate buffer; based on the total mass of the composition (100%), the mass percentage of shikonin micelle particles is 5% to 12%, the mass percentage of phenylboronic acid-modified sodium hyaluronate is 1.5% to 3.5%, and the balance is lactate-sodium lactate buffer. Shikonin micelles are nanoparticles assembled from tocopherol polyethylene glycol 1000 succinate and shikonin, wherein the hydrophilic-lipophilic balance value of tocopherol polyethylene glycol 1000 succinate is 13 to 13.5, and the mass ratio of tocopherol polyethylene glycol 1000 succinate to shikonin is 10:1 to 15:

1. Phenylated boric acid modified sodium hyaluronate is sodium hyaluronate grafted with phenylboronic acid groups. The weight-average molecular weight of the sodium hyaluronate is 800 kDa to 1200 kDa, and the grafting degree of phenylboronic acid groups on the sodium hyaluronate backbone is 18 mol% to 22 mol%. The composition The value is 3.8 to 4.2; wherein, the phenylboronic acid group has the chemical activity to form a borate ester complexation reaction with the ortho-dihydroxy structure to form a gel anchoring layer at the interface of the composition; in an environment with a hydrogen peroxide concentration of 0.1 mmol / L to 1.0 mmol / L, the phenylboronic acid group undergoes an oxidative deboronization reaction and is converted into phenolic hydroxyl groups, causing the gel anchoring layer to undergo a phase transition from a gel state to a sol state to release shikonin.

2. The shikonin composition for inhibiting non-specific inflammation of the reproductive system according to claim 1, characterized in that, The average particle size of the shikonin micelles is 35 nm to 60 nm; the hydrophobic core formed by tocopherol polyethylene glycol 1000 succinate physically encapsulates the shikonin, thus spatially isolating the naphthoquinone core of the shikonin from the aqueous environment in the lactate-sodium lactate buffer solution.

3. The shikonin composition for inhibiting non-specific inflammation of the reproductive system according to claim 1, characterized in that, The shear yield stress of phenylboronic acid-modified sodium hyaluronate in the composition system is greater than 50 Pa; when hydrogen peroxide penetrates into the gel anchoring layer, the oxidative deboronization reaction causes the crosslinking points in the gel anchoring layer to break.

4. The shikonin composition for inhibiting non-specific inflammation of the reproductive system according to claim 1, characterized in that, The lactate-sodium lactate buffer solution contains lactate at a concentration of 0.1 mol / L to 0.3 mol / L, and is used to initially prepare the composition system. The value remained between 3.8 and 4.2, and after the crosslinking point broke, it maintained the hydrogen ion concentration in the local microenvironment.

5. The shikonin composition for inhibiting non-specific inflammation of the reproductive system according to claim 1, characterized in that, Shikonin micelles contain active components that inhibit cell membrane efflux pumps; after the cross-linking points break, the shikonin micelles physically occupy the P-glycoprotein binding sites on the cell membrane through their tocopherol polyethylene glycol 1000 succinate component.

6. The shikonin composition for inhibiting non-specific inflammation of the reproductive system according to claim 1, characterized in that, The phenylboronic acid group is a chemical structure formed by the amidation reaction of aminophenylboronic acid with the carboxyl group on the main chain of sodium hyaluronate; the molar ratio of condensing agent to carboxyl group in the amidation reaction is 1.2:1 to 1.5:

1.

7. The shikonin composition for inhibiting non-specific inflammation of the reproductive system according to claim 1, characterized in that, The composition has a dynamic viscosity of 1500 mPa·s to 3500 mPa·s at 25°C; phenylboronic acid modified sodium hyaluronate forms physical crosslinking points composed of phenylboronic acid groups in the composition system through the coupling of weight-average molecular weight and grafting degree.

8. The shikonin composition for inhibiting non-specific inflammation of the reproductive system according to claim 1, characterized in that, The preparation method of shikonin micelle particles includes: step S91: shikonin and tocopherol polyethylene glycol 1000 succinate are co-dissolved in anhydrous ethanol, and the anhydrous ethanol is removed by rotary evaporation under vacuum conditions of 40°C to 45°C to form a film; step S92: lactate-sodium lactate buffer is added to the film formed in step S91, and hydration is carried out at 55°C to 60°C.

9. The shikonin composition for inhibiting non-specific inflammation of the reproductive system according to claim 1, characterized in that, The composition is in the form of a gel or suppository; the composition is used to repair damaged physical barriers by physically covering the target surface with sodium hyaluronate in response to changes in hydrogen peroxide concentration.

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