A chiral imitated virus nanomedicine carrier for efficiently overcoming gastrointestinal physiological barrier and a construction method thereof

Abstract

This invention discloses a highly efficient chiral viral-like nanomedicine carrier for overcoming the gastrointestinal physiological barrier and its construction method, wherein the chiral viral-like nanomedicine carrier is a chiral viral-like mesoporous silica drug-loaded carrier. The construction method of the highly efficient chiral viral-like nanomedicine carrier for overcoming the gastrointestinal physiological barrier includes: using indomethacin as a model drug, and loading indomethacin IMC into a series of mesoporous silica spherical MSNs, viral mesoporous silica VSNs, and chiral viral-like mesoporous silica CVSNs. The chiral viral-like mesoporous silica of this invention has good degradability and biosafety. Its viral topology and chiral modification can enhance the surface and interfacial activity of the carrier and exhibit certain nanoscale chiral recognition capabilities, showing significant advantages in various processes of crossing the intestinal barrier. It can enhance its functionality as an oral drug carrier, has important research value and clinical translation prospects, and provides a reference for the surface and interfacial design of future drug carriers.

Description

Technical Field

[0001] This invention relates to the field of biomedical technology, and in particular to a chiral viral nanomedicine carrier that efficiently overcomes the physiological barrier of the gastrointestinal tract and its construction method. Background Technology

[0002] Oral administration is currently the preferred method of drug delivery due to its advantages such as good patient compliance, safety, non-invasiveness, and convenience. For nanoparticles to be fully utilized in the body, it is crucial to avoid rapid excretion. For oral formulations, when drugs or nanoparticles reach the gastrointestinal tract, they face numerous obstacles, including the acidic environment of the stomach, intestinal mucus, the epithelial cell barrier, and the downward air and fluid flow caused by intestinal peristalsis. These barriers must be overcome before absorption into the bloodstream, resulting in low bioavailability and poor therapeutic efficacy. The mucus barrier and the epithelial cell barrier are the main reasons hindering the absorption of orally administered nanoparticles into the bloodstream.

[0003] The intestinal mucosal epithelium contains numerous goblet cells. Intestinal mucus, composed of water, mucin, carbohydrates, lipids, and other substances secreted by these goblet cells, forms the intestine's first line of defense against pathogens and endogenous stimuli. The mucus layer, a viscoelastic 3D network structure, can effectively capture and adsorb nanoparticles. However, these nanoparticles are quickly expelled along with the rapid turnover of mucus, resulting in a short retention time for them in the body, rendering them ineffective. This makes the mucus layer a significant obstacle to the absorption of nanoparticles in the gastrointestinal tract.

[0004] The intestinal epithelial cell barrier is formed by the tight junctions between intestinal epithelial cells, goblet cells, M cells, and columnar epithelial cells. In addition to nutrient absorption and metabolism, intestinal epithelial cells also restrict the invasion of harmful substances and infection, making them a major barrier affecting the absorption of orally administered drugs. Intestinal epithelial cells constitute the majority of the intestinal epithelial cell barrier. The intestinal epithelial cell membrane is composed of a lipid bilayer; this unique structure prevents many drugs from directly crossing the cell membrane. For example, large molecules must enter the epithelial cells via receptor-mediated endocytosis. Summary of the Invention

[0005] In view of this, the present invention discloses a chiral viral nanoparticle drug carrier that efficiently overcomes the physiological barrier of the gastrointestinal tract and its construction method, so as to overcome the problem that the intestine, the main barrier to oral drug absorption, greatly limits the oral transport efficiency of drugs and nanoparticles while maintaining the balance of the intestinal environment, resulting in low bioavailability.

[0006] The technical solution provided by the present invention is as follows: Firstly, the present invention provides a chiral viral-like nanomedicine carrier that efficiently overcomes the physiological barrier of the gastrointestinal tract, wherein the carrier is a chiral viral-like mesoporous silica drug-carrying carrier.

[0007] On the other hand, the present invention also provides a method for constructing a chiral viral nanomedicine carrier that efficiently overcomes the physiological barrier of the gastrointestinal tract, comprising: selecting indomethacin as a model drug and loading indomethacin IMC into a series of mesoporous silica spherical mesoporous silica MSNs, viral mesoporous silica VSNs, and chiral viral mesoporous silica CVSNs.

[0008] Further, specifically including: preparing drug-loaded carriers using a solvent evaporation method: first, prepare an IMC acetone solution with a concentration of 10 mg / ml; weigh 60 mg of MSN, VSN, and CVSN samples and add them to a brown bottle; then add 2 ml of a 10 mg / ml indomethacin acetone solution to make the drug / carrier ratio 1 / 3 w / w; seal, and after sonication to homogenize, stir at 300 rpm for 24 h at room temperature; then place the bottle open in a 40°C vacuum oven to evaporate the solvent; after drying, obtain IMC@MSN, IMC@VSN, and IMC@CVSN drug-loaded carriers.

[0009] Further, the synthesis method of the MSNs is as follows: In a solution containing 100 mL of deionized water and 30 mL of ethanol, 1 g of CTAB and 1 mL of ammonia water are added sequentially. After the CTAB is completely dissolved, 3 mL of TEOS is added dropwise. The mixture is stirred at room temperature for 4 hours, then allowed to stand for 24 hours. The product is collected by centrifugation and washed multiple times with water and ethanol. The sample is then dried in a vacuum at 45 °C for 8 hours. Finally, it is calcined in a muffle furnace at 550 °C for 6 hours to remove the CTAB, thus obtaining the MSNs.

[0010] Further, the synthesis method of the viral mesoporous silica VSNs is as follows: First, add 40 ml of deionized water, 1 g of CTAB and 0.18 g of TEA to a round-bottom flask. After stirring the flask in a 60°C water bath for 2 h, add dropwise 20 ml of a cyclohexane-TEOS mixture, 16 ml of cyclohexane and 4 ml of TEOS. After stirring in a 60°C water bath for 48 h, transfer the mixture to a 98°C oil bath and continue stirring for 24 h. Centrifuge to collect the product and wash it several times with water and ethanol. Then, dry the sample in a vacuum at 45°C for 8 h, and then calcine it in a muffle furnace at 550°C for 6 h to remove CTAB, thus obtaining VSNs.

[0011] Furthermore, the synthesis method of chiral viral mesoporous silica CVSNs is as follows:

[0012] Synthesis of AVSNs: VSNs were modified by surface amination with APTES. 100 mg of VSNs after template removal was weighed and ultrasonically dispersed in anhydrous ethanol. 300 μl of APTES was added and the mixture was stirred in a water bath at 50 °C for 24 h. The mixture was washed with anhydrous ethanol several times, centrifuged, dried and the sample was collected to obtain amino-functionalized VSNs, i.e., AVSNs.

[0013] Chiral surface modification was achieved by grafting chiral L / D-alanine via acylation. 20 mg of AVSNs were dispersed in 5 ml of anhydrous DMSO. In the presence of EDCI / HOBT, 81.27 mg of Boc-L / D-alanine was added to the mixture, stirred at room temperature for 48 h, centrifuged, washed alternately with water and alcohol, dried, and the sample was collected to obtain Boc-L / D-AVSNs.

[0014] To remove the Boc protecting group, trifluoroacetic acid (TFA) was used for treatment. 15 mg of Boc-L / D-AVSNs was weighed and sonicated in 15 ml of DMSO until homogeneous. 6.53 ml of TFA was added, and the mixture was reacted at room temperature for 3 h. After centrifugation, the mixture was washed with anhydrous ethanol and dried to obtain L / DVSNs, where LVSN is CVSN.

[0015] This invention provides a chiral, virus-like nanomedicine carrier and its construction method that efficiently overcomes the gastrointestinal physiological barrier. By modifying L-alanine with an amide bond, viral-like mesoporous silica is endowed with chirality, increasing its adhesion to organisms and its potential for chiral-specific recognition. This successfully constructs a viral-like chiral mesoporous silica nanomedicine delivery system capable of efficiently crossing the intestinal barrier.

[0016] Chiral viral mesoporous silica exhibits good degradability and biocompatibility. Its viral topology and chiral modification can enhance the surface and interfacial activity of the carrier and demonstrate certain nanoscale chiral recognition capabilities. It shows significant advantages in various processes of crossing the intestinal barrier, which can enhance its functionality as an oral drug carrier. It has important research value and clinical translation prospects, and provides a reference for the surface and interfacial design of future drug carriers.

[0017] VSNs and CVSNs, as carriers, leverage their topological advantages and chiral properties to alter drug distribution in vivo, particularly prolonging drug persistence. Compared to traditional spherical mesoporous silica, they exhibit higher bioavailability, thus demonstrating superior anti-inflammatory pharmacodynamics. This provides a new approach to addressing common problems of poorly soluble drugs, such as low dissolution, poor absorption, and low bioavailability.

[0018] It should be understood that the above general description and the following detailed description are exemplary and explanatory only, and do not limit the disclosure of the present invention. Attached Figure Description

[0019] The accompanying drawings, which are incorporated in and form part of this specification, illustrate embodiments consistent with the invention and, together with the description, serve to explain the principles of the invention.

[0020] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, for those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0021] Figure 1 These are the series of TEM, SEM, and particle size analysis images of silicon nanoparticles as described in the embodiments of this invention;

[0022] Figure 2 (A) Infrared spectrum, (B) Circular dichroism chromatogram and (C) Thermogravimetric analysis curve of the series of silicon nanoparticles provided in the embodiments of the present invention;

[0023] Figure 3 (A) Nitrogen adsorption curve, (B) BJH pore size distribution curve and (C) SAXS spectrum of a series of silicon nanoparticles provided for embodiments of the present invention;

[0024] Figure 4 The series of mesoporous silicon surface and interface characteristics provided in the embodiments of the present invention include (A) initial contact angle, (B) contact angle variation over time, and (C) Zeta potential.

[0025] Figure 5 The amino acid adsorption characteristics provided in the embodiments of the present invention are shown in (A) amino adsorption CD spectrum and (B) schematic diagram of amino acid adsorption amount.

[0026] Figure 6 The degradation rates of a series of carriers provided in the embodiments of the present invention in different simulated liquids are shown in (A) simulated gastric juice, (B) simulated intestinal juice, and (C) simulated body fluid.

[0027] Figure 7 The series of carrier hemolytic test results provided in the embodiments of the present invention include (A) hemolysis photographs and (B) hemolysis rate;

[0028] Figure 8 The series of carriers provided in the embodiments of this invention adsorb BSA protein;

[0029] Figure 9 The series of vector cytotoxicities provided in the embodiments disclosed in this invention;

[0030] Figure 10 The following are weight gain curves after oral administration of a series of carriers provided in the embodiments of the present invention;

[0031] Figure 11 In vivo toxicity blood biochemical tests of a series of carriers provided in the embodiments of the present invention;

[0032] Figure 12In vivo toxicity blood routine indicators of a series of carriers provided in the embodiments of the present invention;

[0033] Figure 13 These are tissue sections of major organs from control and orally administered vector mice provided in the embodiments of the present invention.

[0034] Figure 14 The series of carriers disclosed in this invention provide intestinal adhesion (A) in vivo imaging and ROI intensity (B) intestinal adhesion rate;

[0035] Figure 15 A three-dimensional scanning image of mucus penetration provided in an embodiment of the present invention;

[0036] Figure 16 The series of carriers disclosed in the embodiments of the present invention provide the following gastrointestinal retention capacity: (A) in vivo imaging of intestinal retention; (B) relative fluorescence intensity of the stomach; and (C) relative fluorescence intensity of the small intestine.

[0037] Figure 17 In vivo imaging distribution and relative fluorescence intensity of a series of carriers provided in the embodiments of the present invention in mouse heart, liver, spleen, lung, kidney and brain;

[0038] Figure 18 The absorption of nanoparticles by intestinal tissue 2 hours after oral administration, as provided in the embodiments of the present invention;

[0039] Figure 19 The ultraviolet absorption spectra of IMC in different media provided in the embodiments of the present invention;

[0040] Figure 20 (A) Nitrogen adsorption-desorption curve and (B) pore size distribution of the drug-loaded carrier provided in the embodiments of the present invention;

[0041] Figure 21 Infrared spectra of the carrier before and after drug loading provided in the embodiments of the present invention;

[0042] Figure 22 XRD patterns of IMC, blank carrier, drug-loaded carrier and IMC carrier mixture provided in the embodiments of the present invention;

[0043] Figure 23 DSC spectra of IMC, blank carrier, drug-loaded carrier, and IMC carrier mixture provided in the embodiments of the present invention;

[0044] Figure 24 The wettability of the drug-loaded carrier provided in the embodiments of the present invention is shown in (A) initial contact angle diagram and (B) contact angle variation trend over time.

[0045] Figure 25Release curves of IMC in different dissolution media provided for embodiments of the present invention: (A) pH 6.5, (B) pH 6.8, (C) pH 7.4;

[0046] Figure 26 The time-drug concentration curves of IMC, IMC@MSN, IMC@VSN, and IMC@CVSN provided in the embodiments of the present invention;

[0047] Figure 27 The in vivo distribution of IMC, IMC@MSN, IMC@VSN and IMC@CVSN provided in the embodiments of the present invention at different times (A) 1h (B) 3h (C) 6h;

[0048] Figure 28 The swelling rate-time curves of physiological saline, IMC raw material, IMC@MSN, IMC@VSN, and IMC@CVSN for treating ankle joint swelling in rats are provided in the embodiments of the present invention.

[0049] Figure 29 The appearance of the rat's foot and ankle swelling and the pathological section of the skin tissue are provided in the embodiments of the present invention. Detailed Implementation

[0050] Exemplary embodiments will now be described in detail, examples of which are illustrated in the accompanying drawings. When the following description relates to the drawings, unless otherwise indicated, the same numerals in different drawings denote the same or similar elements. The embodiments described in the following exemplary embodiments do not represent all embodiments consistent with the present invention. Rather, they are merely examples of systems consistent with some aspects of the invention as detailed in the appended claims.

[0051] The intestinal barrier is a major obstacle to the oral absorption of drugs. While maintaining the homeostasis of the intestinal environment, it also greatly limits the oral transport efficiency of drugs and nanoparticles, which is the main reason for their low bioavailability. Viruses have a high degree of invasiveness and have certain advantages in processes such as penetration, adhesion, and invasion. This implementation plan utilizes the advantages of viral morphology combined with the physicochemical properties of mesoporous silica to construct a drug delivery system that leverages its structural advantages to efficiently cross physiological barriers. Furthermore, since endogenous substances such as DNA and amino acids that constitute living organisms often have chiral properties, the biological behavior of mesoporous silica in vivo is improved by modifying the chiral small molecule L-alanine, increasing its adhesion to biological interfaces and its potential for chiral-specific recognition, thus constructing a virus-like chiral mesoporous silica imaging drug delivery nanosystem aimed at overcoming the intestinal barrier.

[0052] Example 1

[0053] To verify the advantages of viral mesoporous silica morphology as a drug carrier, this embodiment simultaneously synthesized smooth solid silica nanoparticles (SSNs), spherical mesoporous silica (MSNs), viral mesoporous silica (VSNs), and chiral viral mesoporous silica (CVSNs). The synthesis methods are as follows:

[0054] Synthesis of SSNs: 2 mL of LTEOS was added dropwise to a solution containing 40 mL of ethanol, 3 mL of deionized water, and 0.8 mL of ammonia. The mixture was stirred at room temperature for 12 h, and the product was collected by centrifugation. The product was washed several times with water and ethanol. Then, the sample was dried in a vacuum at 45 °C for 8 h.

[0055] Synthesis of MSNs: In a solution containing 100 mL of deionized water and 30 mL of ethanol, 1 g of CTAB and 1 mL of ammonia were added sequentially. After the CTAB was completely dissolved, 3 mL of TEOS was added dropwise. The mixture was stirred at room temperature for 4 h, then allowed to stand for 24 h. The product was collected by centrifugation and washed several times with water and ethanol. The sample was then dried in a vacuum at 45 °C for 8 h. Finally, it was calcined in a muffle furnace at 550 °C for 6 h to remove the CTAB, yielding the final sample.

[0056] Synthesis of VSNs: First, 40 ml of deionized water, 1 g of CTAB, and 0.18 g of TEA were added to a round-bottom flask. The flask was placed in a 60°C water bath and stirred for 2 h. Then, a mixed solution of 20 ml of cyclohexane and TEOS (16 ml cyclohexane, 4 ml TEOS) was added dropwise. After stirring in a 60°C water bath for 48 h, the mixture was transferred to a 98°C oil bath and stirred for another 24 h. The product was collected by centrifugation and washed several times with water and ethanol. The sample was then dried in a vacuum at 45°C for 8 h. Finally, it was calcined in a muffle furnace at 550°C for 6 h to remove CTAB, yielding the final sample.

[0057] Synthesis of AVSNs: The VSNs prepared above were modified by surface amination using APTES. 100 mg of VSNs (after removing the template) was weighed and ultrasonically dispersed in anhydrous ethanol. After adding 300 μl of APTES, the mixture was stirred in a water bath at 50 °C for 24 h, washed repeatedly with anhydrous ethanol, centrifuged, dried, and the sample was collected to obtain amino-functionalized VSNs, i.e., AVSNs.

[0058] Synthesis of L / DVSNs: Chiral surface modification was achieved by grafting chiral L / D-alanine via acylation. 20 mg of AVSNs were dispersed in 5 mL of anhydrous DMSO. In the presence of EDCI / HOBT, 81.27 mg of Boc-L / D-alanine was added to the mixture, stirred at room temperature for 48 h, centrifuged, washed alternately with water and alcohol, dried, and the sample was collected to obtain Boc-L / D-AVSNs.

[0059] To remove the Boc protecting group, trifluoroacetic acid (TFA) was used for treatment. 15 mg of Boc-L / D-AVSNs was weighed and sonicated in 15 ml of DMSO until homogeneous. 6.53 ml of TFA was added, and the mixture was reacted at room temperature for 3 h. After centrifugation, the mixture was washed with anhydrous ethanol and dried to obtain L / DVSNs, where LVSN is CVSN.

[0060] Appropriate amounts of SSNs, MSNs, VSNs, and CVSNs powder samples were ultrasonically dispersed in anhydrous ethanol, coated onto a carbon-coated microgrid copper sheet, dried in an oven, and analyzed by transmission electron microscopy (TEM). The carrier powder was then sputter-coated with gold and analyzed by scanning electron microscopy (SEM). Separately, appropriate amounts of SSNs, MSNs, VSNs, and CVSNs powder were ultrasonically dispersed in anhydrous ethanol, and particle size distribution and particle size were analyzed using DLS.

[0061] The TEM, SEM, and particle size analysis results of the above series of silicon nanoparticles are as follows: Figure 1 As shown.

[0062] SSNs are monodisperse, smooth, non-porous nanospheres; MSNs are monodisperse, rough, mesoporous spheres; and VSNs are monodisperse, virus-like mesoporous carriers with a large number of uniformly distributed vertical nanotubes on their surface. CVSNs also retain their virus-like morphology after grafting with the chiral molecule L-alanine. All four types exhibit uniform morphology and are approximately the same size, with particle size statistically ranging from 90 to 100 nm. However, due to the influence of hydration, the hydrated particle size measured by DLS is slightly larger than the true value, ranging from 130 to 140 nm.

[0063] A series of infrared spectra, circular dichroism chromatograms, and thermogravimetric analysis curves of silicon nanoparticles are shown below. Figure 2 As shown.

[0064] CVSNs infrared spectrum at 1681 cm⁻¹ -1 The retention of the carbonyl stretching vibration peak of the amide group at this location indicates successful grafting of the chiral L-alanine molecule. Furthermore, the infrared spectrum of CVSNs shows a peak at 462 cm⁻¹. -1 The peak of Si-O-Si bending vibration is shown at 1087 cm⁻¹. -1 The peaks show Si-O-Si asymmetric stretching vibrations. L-alanine-grafted VSNs and D-alanine-grafted VSNs exhibit positive and negative Cotton effects in the 210–250 nm range, respectively, while no obvious chiral response signal was observed in VSNs, suggesting successful alanine chiral modification. The weight losses of SSNs, MSNs, VSNs, and CVSNs in the 25–700 °C range were 1.20%, 4.98%, 9.76%, and 24.07%, respectively. This suggests that the grafting amount of the chiral small molecule L-alanine was approximately 14.31%, indicating successful grafting of the chiral small molecule specific to CVSNs.

[0065] Nitrogen adsorption curves, BJH pore size distribution curves, and SAXS spectra of MSNs and VSNs before and after template removal are shown in the figure. Figure 3 As shown.

[0066] MSNs, VSNs, and CVSNs all exhibited distinct Type IV adsorption / desorption isotherms and contained hysteresis loops, indicating the presence of a uniform mesoporous structure. Pore size measurements showed that all three possessed mesoporous channels, with specific values ​​shown in Table 1. The pore sizes were 3.2 nm, 2.9 nm, and 2.5 nm, respectively. Solid, non-porous SSNs did not show a Type IV adsorption / desorption isotherm and lacked mesoporous pore size distribution. SAXS results within the 0.1°–5° (2θ) testing range indicated that, compared to before template removal, VSNs and MSNs exhibited higher diffraction peaks after template removal, suggesting a higher degree of pore structure order.

[0067] Table 1. Specific surface area, pore volume, and pore size of a series of silicon nanoparticles.

[0068]

[0069]

[0070] Surface and interface properties of a series of silicon nanoparticles: The Zeta potentials of SSNs, MSNs, VSNs, AVSNs, and CVSNs were 2.67±1.11, -2.36±0.19, -13.67±4.06, 35.15±3.6, and 21.16±1.36 mV, respectively. SSNs and MSNs exhibited similar surface characteristics and were electrically neutral; VSNs contained a weak negative charge; the potential of AVSNs modified with amino groups increased significantly, showing positive charge, indicating successful amino modification; the surface potential of CVSNs increased compared to VSNs, proving successful chiral molecule grafting. The initial contact angles of SSNs, MSNs, VSNs, and CVSNs were 43.53°, 30.56°, 19.70°, and 14.45°, respectively, and the contact angles of VSNs and CVSNs dropped to 0° in a very short time, proving that both viral topology and chiral grafting can significantly increase the roughness of the vector surface and interface.

[0071] Amino acid adsorption characteristics of chiral viral silicon: The results of amino acid adsorption characteristics are as follows Figure 5 As shown, the CD spectra of viral mesoporous silica with different configurations and chiral modifications exhibit corresponding symmetrical Cotton effects in the adsorption experiments of L / D alanine. It can be concluded that LVSNs have a greater adsorption capacity for L-alanine than for D-alanine; similarly, DVSNs have a greater adsorption capacity for D-alanine than for L-alanine.

[0072] In this embodiment, the rough, spinous interface of VSNs has more contact sites and a larger external surface area compared to spherical carriers, resulting in higher interfacial reactivity. Compared to SSNs and MSNs, VSNs, due to their outer clusters being assembled from monodisperse micelles, have smaller pore sizes. According to the Kelvin equation, the capillary action within these pores is stronger, generating greater additional pressure on the liquid and significantly reducing the water contact angle. Secondly, VSNs exhibit high interfacial affinity and a multi-site rough topology, facilitating wetting and promoting mass transfer of the liquid into the carrier. The adsorption capacity of the carrier interface is also significantly enhanced, further promoting cellular uptake or signal transduction and enhancing its contact, adhesion, and other biological behaviors, thus benefiting the in vivo application of VSNs as drug carriers. Furthermore, amino acid chiral modification enhances the hydrophilicity of the carrier surface, further reducing the contact angle of CVSNs compared to VSNs. CVSNs exhibit selective adsorption properties, showing significantly stronger adsorption capacity for L-amino acids than D-amino acids, demonstrating a certain degree of chiral recognition ability at the nanoscale interface.

[0073] Example 2

[0074] Degradation of the above series of carriers

[0075] The degradation trends of the series of vectors in SGF, SIF and SBF are as follows: Figure 6 As shown.

[0076] As shown in the figure, the series of carriers underwent slow degradation in all three simulated liquids. The degradation rate was relatively fast in the first week, and then slowed down in the following 2-12 weeks. Different pH values ​​of the simulated degradation liquids significantly affected the degradation process, with the degradation rate in the following order: SBF > SIF > SGF. Furthermore, VSN exhibited a faster degradation rate than SSN and MSN; indicating that the chiral-modified CVSN showed a further improvement in degradation rate compared to VSN.

[0077] Example 3

[0078] Hemolytic activity of a series of vectors

[0079] Hemolytic images and hemolysis rate even if the results are as follows Figure 7 As shown.

[0080] The results showed that the hemolysis rates of the series of silicon nanocarriers were concentration-dependent, increasing with increasing concentration. At a concentration of 100 μg / ml, the hemolysis rate of MSN exceeded 10%. In contrast, the hemolysis rates of SSN, VSN, and CVSN only increased slightly with increasing concentration, remaining below 7% in the concentration range of 5–100 μg / ml, exhibiting low hemolysis rates and good blood compatibility, all meeting the national standards for hemolysis of biomaterials. Furthermore, L-alanine chiral modification slightly reduced the hemolysis rate.

[0081] Example 4

[0082] The adsorption results of BSA protein in the series of vectors are as follows: Figure 8 As shown. The weaker the adsorption of protein by the material, the less impact it has on disrupting the balance of anticoagulant / procoagulant factors in the blood, and the higher its safety. The adsorption of BSA by the above carriers showed that the adsorption amounts of SSNs (1.6%), MSNs (8.1%), VSNs (6.7%), and CVSNs (9.5%) were all below 10%, indicating that the carriers are not sensitive to proteins in the blood and have good biocompatibility. Compared to MSNs (8.1%), the adsorption amount of BSA by VSNs was significantly lower. This is because the negatively charged surface of VSNs electrostatically repels the negative charge carried by the protein. The increased protein adsorption by CVSNs compared to VSNs may be due to an increased surface potential or the increased affinity for BSA after surface modification with L-alanine, resulting in higher protein adsorption.

[0083] Example 5

[0084] Cytotoxicity of a series of vectors, such as Figure 9 As shown, the results indicated that after culturing cells with the vectors for 24, 48, and 72 hours, respectively, the SSNs, MSNs, VSNs, and CVSNs vectors at concentrations ranging from 0.1 to 100 μg / ml had no significant effect on the survival rate of Caco-2 cells. Even at concentrations far exceeding the clinically used dose of 100 μg / ml, cell survival remained above 80%. Within the study dosage range, cell survival was not significantly dependent on the vector dosage. These results demonstrate that the aforementioned vectors exhibit good biocompatibility, do not produce significant cytotoxicity to Caco-2 cells, and have no significant difference in their effect on cell survival.

[0085] Example 6

[0086] In vivo toxicity experiments of a series of vectors—weight gain: The weight gain curve of mice after oral administration is shown in the figure below. Figure 10 As shown in the figure. Experimental results indicate that throughout the entire experimental period (including the initial week of adaptation), none of the mice exhibited any abnormal clinical behavior. The weight gain in the blank control group was 36.4%, while the weight gains in the SSN, MSN, VSN, and CVSN groups were 39.1%, 32.1%, 39.3%, and 38.2%, respectively. These results demonstrate that the carrier materials in each group did not affect the weight gain of the mice, and there was no significant difference in weight gain between the experimental groups and the blank control group.

[0087] Example 7

[0088] The in vivo toxicity tests of the series of vectors—blood biochemical indicators and blood routine results are as follows: Figure 11 and Figure 12 As shown. After 15 days of continuous administration, all blood routine indicators were within the reference range of ICR mice, and there were no significant differences in the indicators between the experimental group and the control group, indicating that SSNs, MSNs, VSNs and CVSNs carriers do not cause abnormalities in blood components within the test range. Figure 2-9 The results showed that blood biochemical parameters after administration were all within the reference range for ICR mice, and there were no significant differences in any indicators between the experimental and control groups, indicating that the carriers did not cause damage to liver or kidney function within the test range. In vivo safety experiments showed that SSNs, MSNs, VSNs, and CVSNs carriers have good biocompatibility and low toxicity, meeting the basic requirements for the clinical application of biomaterials.

[0089] Example 8

[0090] In vivo toxicity experiments of a series of vectors—organ-to-body ratio and tissue sections. The results of the organ-to-body ratio are shown in Table 2. The organ and tissue section results of the series of vectors are as follows. Figure 13 .

[0091] Table 2 shows the ratio of in vivo toxicity to organoid toxicity in a series of carriers.

[0092]

[0093] After organ harvesting, no organ enlargement or abnormal coloration was found. The organ-to-body weight ratio of mice in the series of vector sample groups and the blank group showed no significant difference and remained within the normal range for ICR mice. Histological sections also revealed no local inflammation, damage, or necrosis. This indicates that SSNs, MSNs, VSNs, and CVSNs all possess good biocompatibility.

[0094] The above embodiments respectively relate to the degradability, cytotoxicity, and in vitro and in vivo toxicity of a series of carriers. In the physiological environment, the carrier is affected by the action of water molecules, as well as conditions such as ions, enzymes, and temperature, resulting in phenomena such as loss of integrity, change in properties, or reduction in performance. At the same time, due to the abundant silanol groups on the surface of nanosilicon forming hydrogen bonds with water molecules in the degradation solution, it promotes the fracture of the framework, dissolution of the material, and the weight loss gradually increases with the degradation time, and the carrier material degrades. The results of the degradation experiment show that the degradation rate is SSNs < MSNs < VSNs. This may be due to the influence of the morphological characteristics of VSNs. The rough spines on the surface of VSNs endow it with a large number of contact sites, an active interface property. And according to the above description, VSNs have better wettability, a smaller contact angle when contacting the degradation medium, a larger contact area with the liquid, and a faster degradation rate, but it can still maintain the functionality of the carrier in various simulated media. In addition, VSNs have smaller pore sizes, a stronger Kelvin effect, and generate a greater additional pressure on the wetting liquid, which promotes the mass transfer of the degradation solution into the interior of the carrier. In addition to the structure of the material itself, the degradation process is also affected by the pH of the degradation medium. Whether it is SSNs, MSNs or VSNs, the degradation rate increases with the increase of pH. The degradation rate in gastric juice is slower, which does not damage the rigid structure of the carrier and is conducive to protecting the drug for oral absorption in the small intestine part.

[0095] In summary, viral silica degrades slowly in body fluids, providing effective physical protection for the loaded drugs during transport and release. After completing its mission, the carrier can completely degrade, avoiding the safety risks caused by prolonged carrier retention in the body. Regardless of the administration method (oral or injectable), nanocarriers inevitably come into contact with blood; therefore, blood safety is a crucial indicator for evaluating nanomaterials. In hemolytic experiments, despite the rough, spiked surface of VSNs, they exhibited low hemolytic activity (<5%) at clinical concentrations (5-100 μg / mL). Furthermore, L-alanine modification enhanced the carrier's hydrophilicity, thus shielding its surface from reactive oxygen species (ROS) and reducing hemolysis. The hydrophilic silanol groups on the surface of mesoporous silica carriers can bind to albumin through hydrogen bonding, affecting albumin's anticoagulant effect and potentially leading to coagulation or thrombosis. In the protein adsorption experiment, the protein adsorption amounts of SSNs, MSNs, VSNs, and CVSNs were all below 10%, with CVSNs showing a slightly higher adsorption amount than VSNs. This may be because surface modification with L-alanine increased their affinity for proteins. In the cytotoxicity experiment, the cell viability after co-culturing the series of carriers with cells was not significantly dependent on culture time or carrier concentration, and no cytotoxicity was observed. In vivo toxicity studies showed that no abnormal clinical manifestations occurred in the animals throughout the experiment. There was no significant difference in body weight gain among different carriers. After organ collection (heart, liver, spleen, lung, and kidney), no obvious swelling or inflammation was observed, and the organ-to-body ratio was within the normal range. Histological examination showed that the structural characteristics of each organ were clear, the tissue morphology was clear, and no structural changes, local inflammation, or necrosis were observed. There were no significant differences between the groups. There were no significant differences in blood routine test indicators and liver and kidney function biochemical test results among the groups, and the measured values ​​were all within the normal reference range.

[0096] The in vitro toxicity of the vectors was assessed through blood safety and cytotoxicity tests, while in vivo toxicity was assessed by recording clinical manifestations, body weight and organ-to-body ratio, complete blood count, blood biochemical indicators, and tissue sections of important organs. The results indicate that VSNs and CVSNs have good safety profiles as drug carriers both in vitro and in vivo, and can be safely used in the construction of drug delivery systems.

[0097] Example 8

[0098] In vitro intestinal bioadhesion: Results of in vitro intestinal bioadhesion of a series of vectors are as follows... Figure 14As shown in the figure, sufficient contact between the drug carrier and the intestine is a prerequisite for oral absorption. Theoretically, a larger contact area and longer contact time between the carrier and the small intestinal mucosa are beneficial for oral drug absorption. The results in the image show that the adhesion rate of SSN was only about 40% after 5 minutes of elution, while the adhesion rate of MSN was slightly higher than that of SSN, reaching 55% after the elution process. Among the three carriers with different morphologies, VSNs showed significantly enhanced retention on the intestinal mucosa compared to MSNs and SSNs, with over 80% of the carriers adhering to the intestinal tissue, demonstrating good bioadhesion. Figure B shows that the bioadhesion ability of the carrier is closely related to its morphology, with the adhesion ability of the three carriers in the order of VSNs > MSNs > SSNs. Furthermore, due to better wettability and smaller pore size, CVSNs showed a slightly enhanced adhesion rate compared to VSNs, suggesting that the L-alanine modified on its surface generates a chiral response for recognition.

[0099] Example 9

[0100] Intestinal mucus permeability, the results of intestinal mucus permeability of a series of vectors are as follows: Figure 15 As shown in the figure. The green fluorescence represents the FITC-labeled mucus layer, and the red fluorescence represents the RITC-labeled silicon nanoparticles.

[0101] Example 10

[0102] Intestinal retention, the results of intestinal retention over time for a series of vectors are as follows: Figure 16 As shown in the figure, for SSNs, a weak fluorescence signal was observed in the stomach 1 hour after oral administration, while a strong fluorescence signal was observed in the small intestine. At 2 hours, the fluorescence intensity in the small intestine was significantly reduced compared to 1 hour, and almost completely disappeared at 6 hours. These results indicate that the smooth, non-porous SSNs have a small contact area and low friction in the gastrointestinal tract, resulting in extremely rapid excretion and a short retention time in the intestine. For MSNs, fluorescence signals were detected in both the stomach and small intestine 1 hour after administration; after 2 hours, fluorescence signals were only observed in the intestine, mainly concentrated in the middle and lower parts of the small intestine and the large intestine. At 6 hours, fluorescence signals were only observed in the large intestine, i.e., the intestinal excretion site, rather than the absorption site in the small intestine. At 12 hours, they were basically completely metabolized in the intestine, and no strong fluorescence was observed. In contrast, the fluorescence intensity of VSNs in the small intestine at the corresponding time points was significantly stronger than that of SSNs and MSNs, and a strong fluorescence signal could still be captured 12 hours after administration, indicating that VSNs have a longer retention time in the intestine. For CVSNs, at 1 hour post-drug administration, the fluorescence signal was mainly concentrated in the stomach, with only a weak fluorescence signal in the intestine. As time progressed, CVSNs exhibited stronger fluorescence signal intensity than VSNs at each time point. These results correspond to the bioadhesion experiments of isolated intestinal segments.

[0103] Example 11

[0104] The bulk distribution of a series of silicon nanoparticles is shown in the following figures. Figure 17 As shown in the results, the fluorescence signals of VSNs and CVSNs were higher than those of SSNs and MSNs in almost all organs. For VSNs and CVSNs, the distribution intensity in the heart, lungs, and brain was significantly higher than that in the liver, spleen, and kidneys over time. Combining imaging images and ROI fluorescence intensity analysis, the fluorescence intensity in the lungs and brain was CVSN>VSN>MSN>SSN, suggesting that VSNs and CVSNs have the potential to target the brain and lungs. For CVSNs, the fluorescence intensity in each organ reached its maximum value 1 hour after oral administration, and the fluorescence intensity decreased rapidly in the 1-2 hour range. In summary, VSNs and CVSNs have a significant advantage in in vivo distribution compared to SSNs and MSNs.

[0105] Example 12

[0106] The absorption of the series of carriers by the small intestinal villi, such as Figure 18 As shown in the figure. By observing the distribution of fluorescently labeled nanoparticles in small intestine slices of ICR mice, the ability of nanoparticles to overcome both mucus and small intestinal epithelial cell barriers can be visually reflected: CVSNs>VSNs>MSNs>SSNs.

[0107] The mucus layer and the small intestinal epithelial cell layer constitute the intestinal mucosa of the digestive tract. Images show that SSNs exhibit extremely weak red fluorescence signals, mainly concentrated in the center of the intestinal lumen, but almost no red fluorescence is observed near the small intestinal villi, indicating that SSNs are difficult to retain in the intestine and penetrate the intestinal mucosal barrier. MSNs show slightly enhanced fluorescence intensity and wider distribution at the small intestinal mucosa compared to SSNs, but very few red fluorescently labeled nanoparticles enter the interior of the small intestinal villi, mainly existing in the intervillous spaces. VSNs, compared to SSNs and MSNs, show high fluorescence intensity at the small intestinal mucosa, mainly distributed around the small intestinal villi and capable of entering them. Furthermore, L-alanine-modified CVSNs show uniform red fluorescence distribution around the small intestinal villi with significantly enhanced fluorescence signals, and also exhibit extremely strong fluorescence signals inside the small intestinal villi. These results indicate that VSNs and CVSNs can penetrate the intestinal barrier and be absorbed into the bloodstream through the small intestinal villi.

[0108] For nanocarriers to be fully utilized in vivo, it is crucial to avoid rapid clearance by the organism. Therefore, extending the residence time of drug-carriers at the absorption site is essential. In intestinal adhesion experiments, VSNs and CVSNs exhibited good bioadhesion. The rough spiked surface of viral silica is highly reactive and can provide numerous contact sites to "anchor" biological tissue when it comes into contact with the tissue interface.

[0109] The intestinal mucus layer poses a significant physiological and mechanical barrier for orally administered nanoscale drug carriers. Furthermore, goblet cells in the small intestine continuously secrete new mucus, and intestinal peristalsis promotes its timely renewal and expulsion. Drug carriers that cannot rapidly penetrate to the epithelial cells at the bottom of the mucus layer face elimination. In mucus penetration experiments, SSNs were mainly concentrated in the upper mucus layer near the intestinal lumen, with almost no particles crossing the mucus barrier, indicating that the mucus layer is a crucial barrier limiting the absorption of nanoparticles via oral administration. MSNs showed slightly enhanced distribution in the mucus layer compared to SSNs, and some particles penetrated the mucus to reach the epithelial cell layer, but still failed to achieve effective longitudinal penetration along the z-axis. In contrast, the virus-like morphology of VSNs, with its rough, spiked surface, allows for multi-site contact with the mucus layer, enabling deep diffusion and widespread distribution within the mucus layer. Compared to SSNs and MSNs, VSNs exhibit superior mucus penetration and effectively overcome this barrier. Rough, spiny surfaces act as interfaces for breaking through biological membranes, generating numerous curved recognition sites at the biomembrane junctions. These geometric centers attract the aggregation of corresponding proteins and lipids, further evolving into bioactive centers that mediate cell recognition or signal transduction, positively promoting biological behaviors such as mucosal adhesion and cellular uptake. This also provides greater potential for improving the bioavailability of loaded drugs. Furthermore, VSNs and CVSNs exhibit strong bioadhesion capabilities and longer retention times in vivo. After absorption into the bloodstream via the intestine, different carriers show varying distribution patterns, suggesting that differences in morphology and chiral modifications may lead to different affinities for different organs. VSNs and CVSNs significantly improve the distribution of carriers in the lungs, brain, and kidneys of animals, demonstrating their potential application value in delivering drugs for the treatment of lung, brain, and kidney diseases, and also providing insights into organ-targeted drug release.

[0110] In the small intestinal villus absorption experiment, SSNs were mainly concentrated on the intestinal lumen side. Due to their weak mucus permeability, there was almost no red fluorescence distribution near the villi, indicating that the small intestinal villi have extremely poor absorption of smooth, solid silica spheres. MSNs, compared to SSNs, have a rougher porous surface and are more widely distributed in the intestinal lumen, but mostly distributed in the intervillous spaces; no fluorescence distribution was observed inside the small intestinal villi. VSNs, compared to SSNs and MSNs, showed high fluorescence intensity at the small intestinal mucosa, mainly distributed around the small intestinal villi and capable of penetrating them. These results indicate that the surface roughness of nanoparticles is an important factor affecting their penetration of the intestinal mucosal barrier; that is, virus-like multi-contact nanoparticles have better intestinal mucosal penetration. Furthermore, L-alanine-modified CVSNs showed significant fluorescence distribution both on the small intestinal villus side and inside the villi. CVSNs possess a unique virus-like rough spiked surface and chiral modification, increasing their absorption capacity by the small intestinal villi.

[0111] Example 13

[0112] The drug loading capacity and drug loading rate of the drug-loaded carriers are shown in Table 3.

[0113] Table 3. Drug loading capacity and drug loading efficiency of different drug delivery carriers

[0114]

[0115] Example 14

[0116] State and properties of drugs in carriers

[0117] After being loaded into IMC, the specific surface area, pore volume, and pore size of MSNs, VSNs, and CVSNs all change. Therefore, the specific surface area, pore volume, and pore size of the drug-loaded carrier were tested, and the results are shown in […]. Figure 20 And Table 4.

[0118] The results show that after loading IMC into the carrier, the specific surface area and pore volume of the drug-loaded carrier are significantly reduced compared to the original blank carrier, and the pore size is also significantly reduced. This fully demonstrates that the drug is effectively loaded into the mesoporous channels of the carrier.

[0119] Table 4 Changes in specific surface area, pore volume, and pore size before and after drug loading.

[0120]

[0121] FTIR results of drug delivery carriers are as follows Figure 19 As shown.

[0122] Structural analysis based on peak positions revealed that, compared to the active pharmaceutical ingredient (API) IMC, the drug-loaded carrier exhibited the group characteristic peaks of the IMC API after drug loading, further demonstrating that the IMC was successfully loaded into the pores.

[0123] XRD analysis was performed on a series of samples, including IMC, drug delivery carriers, blank carriers, and physical mixtures of IMC. The results are as follows: Figure 22 As shown.

[0124] The results showed that no crystal diffraction peaks were observed in the empty carrier in the spectra, and all peaks were typically broad, indicating that the carriers were in an amorphous state. In contrast, the IMC spectra showed numerous sharp diffraction peaks, indicating that the drug had a highly crystalline structure. Physical mixtures of IMC with MSNs, VSNs, and CVSNs also exhibited several strong drug diffraction peaks. In contrast, the IMC@MSN, IMC@VSN, and IMC@CVSN drug-loaded carriers showed almost no drug diffraction peaks, indicating that the drug underwent a crystal transformation and existed in an amorphous state after loading.

[0125] DSC analysis was performed on a series of samples, including physical mixtures of IMC, drug delivery carriers, blank carriers, and IMC. The results are as follows: Figure 23 As shown in the figure, the results indicate that IMC exhibits a significant endothermic peak at around 160℃, suggesting that the IMC active pharmaceutical ingredient is in a crystalline state. The physical mixture of IMC and the carrier shows a relatively small endothermic peak at around 160℃, further confirming their crystalline state. No endothermic peaks were observed in the empty carriers MSNs, VSNs, and CVSNs, indicating that they are all in an amorphous state. However, after IMC is loaded into the carrier, it shows almost no endothermic peak at 160℃, indicating that the drug undergoes a crystalline change after being loaded into the carrier and exists in an amorphous state.

[0126] Example 15

[0127] Drug carrier wettability

[0128] The wettability of the drug delivery carrier, such as Figure 24 As shown. Figure 24 The wettability of the drug-loaded carriers is shown in Figures A and B. (A) Initial contact angle and (B) Contact angle variation over time. The initial contact angle of the IMC drug substance decreased significantly after loading into the carrier. Within 30 seconds, the contact angle of the IMC drug substance decreased from 66.09° to 41.57°, the contact angle of IMC@MSN decreased from 35.26° to 14.65°, and the contact angle of IMC@VSN decreased from 27.28° to 5.62°. The contact angle of IMC@CVSN decreased from 23.75° to 0° within 20 seconds. This indicates that loading into the mesoporous silica-constructed drug delivery system can significantly improve the wettability of the drug substance IMC. VSNs and CVSNs, leveraging their surface roughness advantages, showed the strongest improvement in IMC wettability.

[0129] Example 16

[0130] In vitro drug dissolution, the results of drug dissolution and release in vitro are as follows: Figure 25 As shown in the figure, the results indicate that in three different PBS media (pH 6.5, 6.8, and 7.4), the drug-loaded carriers significantly improved the dissolution rate and amount of IMC due to the reduction in particle size of IMC within the nanoscale channels and the change in the crystallization state of the drug (from crystalline to amorphous). In pH 6.5 PBS, the dissolution rate of the active pharmaceutical ingredient (API) IMC reached 77% at 120 min, IMC@MSN reached 94% at 120 min, while IMC@VSN and IMC@CVSN reached 100% at 90 min, consistent with their wettability results. The high interfacial activity of VSNs and CVSNs enhances their interaction with the dissolution medium, promoting their entry into the internal channels of the carrier for drug dissolution. Furthermore, the cumulative release amount and release rate of both the IMC API and the drug-loaded carrier increased with increasing pH.

[0131] Example 17

[0132] The specificity, standard curve, precision, and extraction recovery of the in vivo pharmacokinetic method for IMC were investigated. Results showed that IMC and the internal standard exhibited good peak shapes and complete separation, and endogenous substances in blank plasma did not affect the determination of IMC and the internal standard. The retention time of IMC was 7.94 min, and the retention time of the internal standard was 4.12 min. Therefore, this method has good specificity for the determination of IMC content.

[0133] The standard curve and precision recovery rates are shown in the table. In the equation, y represents the IMC drug concentration, and x represents the peak area ratio of IMC to the internal standard. The results in the table show that the peak area ratio of IMC to the internal standard has a good linear relationship with the concentration, R0. 2 The RSD was >0.99, and the standard curve met the methodological requirements. The RSDs for intra-day and inter-day precision at all concentrations were less than 15%, indicating that the precision of this method met the requirements for biological sample analysis. Furthermore, the extraction recoveries of NMS and IMC also met the requirements for biological sample determination.

[0134] Table 5. Standard curve, accuracy, and extraction recovery rate of IMC in plasma samples.

[0135]

[0136] Example 18

[0137] The results of drug-carrier pharmacokinetic studies, including blood drug concentration-time curves, are as follows: Figure 26 As shown in Table 6, the pharmacokinetic parameters obtained using DAS software are presented.

[0138] Table 6. Pharmacokinetic parameters of the active pharmaceutical ingredient and carrier formulation after oral administration.

[0139]

[0140] From the perspective of time to peak concentration, the time to peak concentration of IMC was significantly shortened after being loaded into a vector. IMC@MSN, IMC@VSN, and IMC@CVSN could shorten the time to peak concentration of raw material IMC from 12 hours to 6, 4, and 8 hours, respectively. From the perspective of peak plasma concentration (C0), the peak concentration of IMC was significantly reduced. max As can be seen, the order of peak plasma concentrations is: IMC@VSN > IMC@CVSN > IMC@MSN > IMC. Under the same dosage conditions, VSN can significantly increase the peak plasma concentration of indomethacin. The relative bioavailability of IMC@VSN is approximately 10 times higher than that of IMC and twice that of IMC@MSN. In addition, due to the presence of chiral response, the bioavailability of IMC@CVSN is still slightly higher than that of IMC@VSN.

[0141] Example 19

[0142] In vivo drug distribution in IMC drug delivery systems, the results of which are as follows: Figure 27 As shown, for the IMC active pharmaceutical ingredient (API), the distribution in the liver consistently increased, while the distribution in other organs decreased to varying degrees after 6 hours. This indicates that the drug distributed to each organ is metabolized relatively quickly. For IMC@MSN, the maximum drug concentration in each organ occurred at 3 hours, with varying degrees of metabolism occurring at 6 hours. After the drug was loaded into the carrier, the drug distribution in each organ increased compared to the API. For IMC@VSN, the drug concentration in each organ almost continuously increased, and the elimination rate was slow. In contrast, the maximum drug concentration in each organ for IMC@CVSN occurred at 3 hours. Compared to IMC@VSN, the concentration in each organ increased rapidly from 1 to 3 hours, after which metabolism in organs such as the heart was extremely rapid. Notably, IMC@VSN was metabolized more slowly, maintaining a high drug concentration in each organ even after 6 hours, and the concentration continued to increase. At 6 hours, each carrier showed the highest distribution among all tested organs.

[0143] Example 20

[0144] The efficacy of the IMC drug delivery system and the trend of rat paw edema rate over time are as follows: Figure 28 As shown in the figure, after 0.5 hours of drug administration, the swelling rate in all groups was above 30%, indicating that the paw edema model was successfully established. The negative saline group showed severe swelling, while the paw edema levels in the positive IMC raw material control group and the drug-loaded formulation group decreased. The paw edema rates of IMC@MSN, IMC@VSN, and IMC@CVSN rats were all lower than those in the positive control group. The results of the decrease in paw edema rate show that the paw edema rates of IMC@VSN and IMC@CVSN decreased to below 10% within 3 hours, and continued to decrease compared to IMC@MSN from 3 to 6 hours, reaching below 5% at 6 hours.

[0145] Morphology and pathological tissue sections of the inflamed area, as shown in the following figures Figure 27 As shown.

[0146] The results showed that, visually, the saline group still exhibited severe ankle swelling at 6 hours, while the swelling in the other groups decreased to varying degrees due to the anti-inflammatory effects of the medication. The saline group showed subcutaneous hemorrhage and edema, with extensive infiltration of inflammatory cells (including neutrophils and lymphocytes). The IMC raw material group showed subcutaneous and muscle tissue hemorrhage, mild edema, and inflammatory cell infiltration. The IMC@MSN group showed subcutaneous inflammatory cell infiltration and mild edema; while the IMC@VSN and IMC@CVSN groups showed only a small amount of mild inflammatory cell infiltration. The severity of the lesion was assessed based on the degree of inflammatory cell infiltration: Saline > IMC > IMC@MSN > IMC@VSN > IMC@CVSN.

[0147] The above embodiments used indomethacin, a poorly soluble BCS class II nonsteroidal drug with simple structure and well-defined efficacy, as a model drug. IMC@MSN, IMC@VSN, and IMC@CVSN drug delivery systems were constructed using a solvent evaporation method with an IMC-carrier mass ratio of 1:3. To study the drug loading capacity and in vitro dissolution of the drug delivery carriers, an in vitro analytical method for IMC was developed. This included standard curves of drug concentration and UV absorbance in methanol and PBS solutions at pH 6.5, 6.8, and 7.4.

[0148] The drug loading capacity of the IMC@MSN, IMC@VSN, and IMC@CVSN drug-loaded systems remained stable at around 20%. Compared with the unloaded empty carrier, the specific surface area, pore volume, and pore size of the drug-loaded carriers all decreased significantly. Due to the presence of the mesoscopic structure, the drug carrier can change the form in which the drug exists, transforming crystalline drugs into an amorphous state. FTIR results showed that the peak positions of the drug-loaded carrier were consistent with those of the blank carrier, and the characteristic peaks of the IMC active pharmaceutical ingredient were hidden. The IMC was loaded into the pores, effectively improving the drug dispersion. In the XRD analysis results, no crystal diffraction peaks were observed in the spectra, and all peaks showed typical broad peaks, indicating that these carriers were in an amorphous state. The IMC spectra of the active pharmaceutical ingredient showed a large number of sharp diffraction peaks, indicating a highly crystalline structure. The physical mixture of IMC and blank carrier both showed several strong drug diffraction peaks. In contrast, the drug-loaded carriers IMC@MSN, IMC@VSN, and IMC@CVSN showed almost no drug diffraction peaks, indicating that the drug had undergone a crystal transformation and existed in an amorphous state after being loaded. DSC analysis results showed that the drug-loaded carrier did not show obvious endothermic peaks compared to the drug and physical mixture. The results of XRD and DSC together proved that the drug underwent a crystal transformation after being loaded into the pores of the carrier

[60] . The carrier has a mesoscopic structure, and the limited nanoscale pore size will restrict the crystallization of the drug and prevent the crystallization conditions, thus dispersing the poorly soluble drug at the molecular or nanocrystalline level.

[0149] The contact angle of the drug-loaded carrier is significantly reduced compared to the active pharmaceutical ingredient (API). This is because, on the one hand, the carrier alters the form in which the drug exists; on the other hand, the carrier surface and pores contain a large number of hydrophilic hydroxyl groups, and the pores generate a capillary effect with the liquid, improving the wetting effect of the drug. Furthermore, the contact angle trends of IMC@MSN, IMC@VSN, and IMC@CVSN are consistent with those of the unloaded carrier.

[0150] In in vitro dissolution experiments, all drug-loaded carriers significantly improved the release rate and extent of drugs compared to IMC. The release rates of the three drug-loaded carriers were generally similar, but IMC@VSN and IMC@CVSN still showed a slight advantage over IMC@MSN. This is because the nanotube structure on the surface further improved the state of the drug, resulting in smaller drug particles and better dispersion.

[0151] In in vivo experiments on laboratory animals, the advantages of VSNs and CVSNs as drug delivery carriers became apparent: Firstly, in pharmacokinetic experiments, with similar in vitro dissolution results for the drug delivery systems, the relative bioavailability of IMC@VSN and IMC@CVSN was approximately 10 times higher than that of IMC and twice that of IMC@MSN. In vivo drug distribution results showed significant differences in drug distribution between IMC and the drug delivery carrier groups in mice, especially with IMC@VSN exhibiting slower metabolism. Drug concentrations in various organs continued to increase from 1 to 6 hours, and at 6 hours, all carriers showed the highest distribution in all tested organs. Pharmacodynamic results matched the pharmacokinetic bioavailability, with the final paw edema rate and tissue inflammation trend showing the order: IMC > IMC@MSN > IMC@VSN > IMC@CVSN. The reasons are as follows: First, the IMC loaded into the vector is in an amorphous state, which has a higher dissolution rate than crystalline IMC; second, the rough outer cluster structure of VSNs may significantly enhance bioadhesion, increase drug absorption, and maintain a high blood drug concentration for a long time, thus fully exerting a long-lasting effect. This preliminary demonstration shows that viral vectors relying on topological structures have a strong delivery advantage.

[0152] This invention successfully synthesizes L-alanine-modified chiral virus-like mesoporous silica with uniform particle size and good dispersibility. While retaining the excellent physicochemical properties of traditional mesoporous silica, it also has excellent wettability and chiral selective adsorption capacity, which provides a reference for the preparation of mesoporous silica for drug delivery.

[0153] The biosafety of a series of silicon-based carriers was evaluated in the above embodiments, including blood compatibility, cytotoxicity, and in vivo toxicity after oral administration. All indicators met the basic requirements for the application of biomaterials. However, hemolytic experiments on high-concentration carriers are lacking. In vivo toxicity should be investigated by administration methods such as tail vein injection to conduct a systematic and comprehensive evaluation of biosafety.

[0154] Chiral viral-like mesoporous silica exhibits excellent intestinal retention and intestinal barrier crossing capabilities. Based on these results, further research should focus on expanding its applications in the biomedical field, such as encapsulating or loading contrast agents like gadolinium nanoparticles to enhance bioimaging.

[0155] Using IMC as a model drug, a chiral virus-like drug delivery system was constructed, significantly improving the in vivo bioavailability of the drug. Given its relatively long in vivo retention time, it can be subsequently loaded with anticancer drugs such as doxorubicin to construct a sustained-release drug system.

[0156] Other embodiments of the invention will readily occur to those skilled in the art upon consideration of the specification and practice of the invention disclosed herein. This application is intended to cover any variations, uses, or adaptations of the invention that follow the general principles of the invention and include common knowledge or customary techniques in the art not disclosed herein. The specification and examples are to be considered exemplary only, and the true scope and spirit of the invention are indicated by the claims.

Claims

1. A method for constructing a chiral viral-mimicking nanomedicine carrier that efficiently overcomes the physiological barrier of the gastrointestinal tract, characterized in that, The carrier is a chiral viral-like mesoporous silica drug-carrying carrier; The construction method includes: selecting indomethacin as a model drug and loading indomethacin IMC into chiral viral mesoporous silica CVSNs; Specifically, the preparation of the drug-loaded carrier using a solvent evaporation method includes: first, preparing a 10 mg / ml IMC acetone solution; weighing 60 mg of CVSN sample and adding it to a brown bottle; then adding 2 ml of a 10 mg / ml indomethacin acetone solution to make the drug / carrier ratio 1 / 3 w / w; sealing the bottle; after ultrasonic homogenization, stirring at 300 rpm for 24 h at room temperature; then placing the bottle open in a 40 ℃ vacuum oven to evaporate the solvent; and finally obtaining the IMC@CVSN drug-loaded carrier. The synthesis method of chiral viral mesoporous silica CVSNs is as follows: Synthesis of AVSNs: VSNs were modified by surface amination with APTES. 100 mg of VSNs after template removal was weighed and ultrasonically dispersed in anhydrous ethanol. 300 μl of APTES was added and the mixture was stirred in a water bath at 50 °C for 24 h. The mixture was washed with anhydrous ethanol several times, centrifuged, dried and the sample was collected to obtain amino-functionalized VSNs, i.e., AVSNs. Chiral modification of the surface was achieved by grafting the chiral molecule L-alanine via an acylation reaction. 20 mg of AVSNs were dispersed in 5 ml of anhydrous DMSO. In the presence of EDCI / HOBT, 81.27 mg of Boc-L-alanine was added to the above mixture. The mixture was stirred at room temperature for 48 h, centrifuged, washed with water / alcohol alternately, dried, and the sample was collected to obtain Boc-L-AVSNs. To remove the Boc protecting group, the product was treated with trifluoroacetic acid (TFA). 15 mg of Boc-L-AVSNs was weighed and sonicated in 15 ml of DMSO until homogeneous. 6.53 ml of TFA was added, and the mixture was reacted at room temperature for 3 h. After centrifugation, the product was washed with anhydrous ethanol and dried to obtain LVSNs, of which LVSN is CVSN.

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