A carrier drug conjugate, its preparation method and application

Abstract

This application belongs to the fields of biomedical technology, nanomedicine, and drug delivery technology, and specifically relates to a non-absorbable carrier drug conjugate, which includes at least the structure shown in Figure 20: wherein component A is a non-absorbable carrier unit; component B is a linker; and component C is a receptor binding unit. The non-absorbable carrier drug conjugate exhibits a hydration kinetic size change of no more than 5% within 24 hours in the physiological environment of the digestive tract, and the proportion absorbed within 24 hours in the physiological environment of the digestive tract is no more than 1%. When administered through the digestive tract, the carrier drug conjugate of this invention remains stable in the digestive tract environment and is not absorbed into the systemic circulation. Therefore, it can specifically bind to target sites on the surface of digestive tract cells, avoiding the limitations of traditional drug administration that targets various parts of the body and reducing side effects.

Description

Technical Field

[0001] This application belongs to the fields of biomedical technology, drug conjugation technology, and drug delivery technology, and specifically relates to a drug conjugate carrier, its preparation method, and its application. Background Technology

[0002] Numerous receptors expressed in the human digestive tract play crucial roles in cell signaling and physiological regulation. For example, activation of free fatty acid receptors (FFA1-4) and glucose-dependent insulinotropic receptors (GPR119) in the intestine can promote the secretion of cholecystokinin and glucose-dependent insulinotropic peptides (Zhao, YF, Front Endocrinol, 2022); activation of bile acid membrane receptors (TGR5) can promote the secretion of glucagon-like peptide-1 (GLP-1) (Lun, W. et al., Acta Pharm Sin B, 2024). Therefore, these targets have gradually become a research hotspot for the treatment of various diseases, including digestive tract diseases, metabolic diseases, and cancer.

[0003] Despite the significant therapeutic potential of drug design targeting these targets, research remains challenging. Because these targets are not specifically distributed in the digestive tract but are widely expressed throughout the body, drugs can enter the bloodstream via passive diffusion or transporter-mediated pathways, modulating targets across the body. This redistribution in vivo can lead to off-target effects and potential side effects, impacting clinical application (Gimeno, RE et al., Cell Metabolism, 2020). For example, studies have shown that long-term use of TGR5 small molecule agonists may lead to excessive accumulation of bile acids in the body, causing toxic side effects such as gallbladder enlargement, liver inflammation, skin itching, nerve inhibition, and even cancer (Portincasa, P et al., Nutrients, 2020; Bertolini, A. et al., Seminars in Immunopathology, 2022; Fuchs, CD and M. Trauner, Nature Reviews Gastroenterology & Hepatology, 2022). In addition, drug redistribution can reduce its effective accumulation at the target site in the digestive tract. Even if some drugs can reach the target site, they may still dissociate after binding to the target site, making it difficult to maintain a lasting effect.

[0004] To overcome these challenges, researchers have increasingly turned to developing drugs with lower absorption rates. By limiting drug absorption and systemic distribution, systemic adverse reactions can be reduced, allowing the drug to act only at its target site. Simultaneously, this can increase drug accumulation at the target site, reduce the dosage, and improve patient tolerability. For example, Cao et al. increased the molecular weight and positive charge of the TGR5 small molecule agonist by introducing a quaternary ammonium group, thereby reducing intestinal absorption and systemic exposure (Cao, H et al., Scientific Reports, 2016). Shen et al. designed the TGR5-DPP4 bifunctional molecule using a strategy of increasing molecular weight, similarly reducing systemic drug exposure (Han, FH et al., European Journal of Medicinal Chemistry, 2022). Furthermore, A. Sloan Devlin et al. found that 7-sulfocholic acid, due to the presence of its polar group, can be restricted to the intestine, reducing off-target effects caused by entering systemic circulation (Chaudhari, SN et al., Nature Chemical Biology, 2021). Ma et al. incorporated linagliptin as a hydrophilic side chain into MN6 ([4-(2,5-dichlorophenoxy)pyridin-3-yl]-(4-cyclopropyl-3,4-dihydro-2H-quinoxolin-1-yl)methane) to develop a poorly absorbed drug OL3 (Ma, SY et al., Acta Pharmacologica Sinica, 2016).

[0005] However, the development of poorly absorbed drugs also faces a series of bottlenecks. First, achieving a balance between low absorption and high agonistic efficacy is a complex challenge; simply reducing absorption often overlooks effectiveness. Phase II clinical trials revealed that the in vivo exposure of GSK's TGR5 agonist SB756050 was not linearly related to dose, exhibiting poor activity with a half-maximal effective concentration (MCD) of only 1.3 μM for the human TGR5 receptor (Hodge, RJ et al., Clin Pharmacol Drug Dev, 2013). Second, it is necessary to consider whether poorly absorbed drugs can maintain their non-absorbable form. Once a drug molecule binds to a receptor, receptor internalization may lead to the bound drug being endocytosed by the cell, entering the cell. Ensuring that all drug molecules are completely unabsorbed during long-term treatment is not easy. Furthermore, in vivo enzyme metabolism may convert the drug into other easily absorbed substances.

[0006] Therefore, developing drugs that have a strong affinity for the target and can continuously interact with the target without being absorbed into the systemic circulation is of great significance for achieving the best therapeutic effect. Summary of the Invention

[0007] This invention aims to provide a class of non-absorbable oral carrier drug conjugates with three significant advantages. First, the non-absorbable nature of these carrier drug conjugates means they are primarily distributed in the digestive tract after administration, making them difficult to absorb into the bloodstream. Therefore, they can specifically bind to target sites on the cell membrane surface of the digestive tract. Compared to traditional receptor binders, these carrier drug conjugates reduce potential systemic adverse reactions by decreasing systemic distribution. Second, upon contact with the cell membrane surface, the carrier drug conjugates induce sparsely distributed receptors on the membrane surface to aggregate towards the conjugate, increasing local receptor concentration and thus enhancing the effect of agonist or inhibitory signaling. Third, the carrier drug conjugates can deliver receptor binders to the cell surface and increase their affinity for the target site.

[0008] According to one aspect of the present invention, a non-absorbable carrier drug conjugate is provided, comprising at least... Figure 20 The structure shown:

[0010] Component A is a non-absorbable carrier unit; component B is a linker; and component C is a receptor binding unit.

[0011] The non-absorbable carrier drug conjugate exhibits a hydration kinetic size change of no more than 5% within 24 hours in the physiological environment of the digestive tract, and the proportion absorbed within 24 hours in the physiological environment of the digestive tract is no more than 1%.

[0012] In some embodiments, the molar ratio of component B to component A is between 1,000 and 5,000,000, and the molar ratio of component C to component A is between 100 and 500,000; preferably, the molar ratio of component B to component A is between 5,000 and 4,000,000, and the molar ratio of component C to component A is between 300 and 400,000; more preferably, the molar ratio of component B to component A is between 10,000 and 4,000,000, and the molar ratio of component C to component A is between 500 and 400,000; most preferably, the molar ratio of component B to component A is between 20,000 and 3,000,000, and the molar ratio of component C to component A is between 2,000 and 300,000.

[0013] In some embodiments, the material of component A is selected from one or a combination of inorganic and organic materials; preferably, the material of component A is selected from one or a combination of silicon dioxide, graphene oxide, carbon quantum dots, carbon nanotubes, hydroxyapatite, titanium dioxide, metal-organic frameworks, covalent organic frameworks, and poly(lactic-co-glycolic acid); more preferably, the material of component A is selected from one or a combination of silicon dioxide, carbon nanotubes, and poly(lactic-co-glycolic acid); most preferably, the material of component A is silicon dioxide.

[0014] In some embodiments, the hydration kinetic dimension of component A is between 400 nm and 10000 nm; preferably, the hydration kinetic dimension is between 400 nm and 2000 nm.

[0015] In some embodiments, the aspect ratio of component A is ≥1.25; preferably, the aspect ratio is ≥1.5; more preferably, the aspect ratio is between 1.5 and 15.0; most preferably, the aspect ratio is between 6.0 and 12.0.

[0016] In some embodiments, the material of component B is selected from hydrophobic or hydrophilic polymers; preferably, the material of component B is selected from hydrophilic polymers; more preferably, the material of component B is selected from polyethylene glycol and its derivatives; most preferably, the material of component B is selected from polyethylene glycol derivatives, wherein one end of the polyethylene glycol molecule is modified to an amino group. In some embodiments, the chain length of component B can be in the range of 5–60 nm, such that after component B is attached to component A, the hydration kinetic size of component A increases by 5–60 nm. In some embodiments, the material of component B is polyethylene glycol and its derivatives containing 5 or more -CH2-CH2-O repeating units; preferably, the material of component B is polyethylene glycol and its derivatives containing 16–24 -CH2-CH2-O repeating units.

[0017] In some embodiments, the material of component C is selected from one or a combination of G protein-coupled bile acid receptor (TGR5) agonists, G protein-coupled receptor 40 (GPR40) agonists, G protein-coupled receptor 120 (GPR120) agonists, G protein-coupled receptor 41 (GPR41) agonists, G protein-coupled receptor 43 (GPR43) agonists, G protein-coupled receptor 119 (GPR119) agonists, calcium ion-sensitive receptor (CaSR) agonists, and G protein-coupled receptor 35 (GPR35) agonists; preferably, the material of component C is selected from TGR5 agonists and GPR 35. Agonist; more preferably, the material of component C is selected from bile acid compounds and their analogues, INT-777 (CAS: 1199796-29-6), zapust (CAS: 37762-06-4), pamoic acid (CAS: 130-85-8), lodusamide (CAS: 53882-12-5), YE120 (CAS: 383124-82-1); even more preferably, the material of component C is selected from cholic acid, deoxycholic acid, chenodeoxycholic acid, ursodeoxycholic acid, lithocholic acid, pamoic acid, etc.; most preferably, the material of component C is selected from deoxycholic acid and pamoic acid. In some embodiments, the dissociation constant of the material of component C in the dissociation reaction with the corresponding receptor is not higher than 10 μM.

[0018] In some embodiments, components A and C are not directly connected; components A and B are connected by covalent bonds or connecting functional groups, and components C and B are connected by covalent bonds or connecting functional groups. Preferably, the connecting functional group connecting components C and B is selected from one or a combination of amide bonds, ester bonds, thioether bonds, triazole bonds, and carbamate bonds. Most preferably, the connecting functional group connecting components C and B is an amide bond.

[0019] In some embodiments, the carrier-drug conjugate of the present invention comprises at least three components: component A, component B, and component C. The molar ratio of component B to component A is between 20,000 and 3,000,000, and the molar ratio of component C to component A is between 2,000 and 300,000. The material of component A is selected from silicon dioxide, and the size of component A is between 400 nm and 2,000 nm. The material of component B is polyethylene glycol and its derivatives, and the material of component C is selected from TGR5 agonists.

[0020] In some embodiments, the carrier-drug conjugate of the present invention has at least the first characteristic or at least two of the following characteristics:

[0021] (1) The carrier drug conjugate is non-absorbable; the non-absorbability means that it is not absorbed by the intestine and does not enter the systemic circulation.

[0022] (2) The carrier drug conjugate has target binding properties; the target includes one or more of the following: bile acid receptor (TGR5), free fatty acid receptor 1 (GPR40), free fatty acid receptor 2 (GPR43), free fatty acid receptor 3 (GPR41), free fatty acid receptor 4 (GPR120), G protein-coupled receptor 119 (GPR119), calcium ion sensitive receptor (CaSR), G protein-coupled receptor 35 (GPR35), and the dissociation constant with the corresponding target is not higher than 10 μM.

[0023] In some embodiments, in the carrier-drug conjugate of the present invention, component B is coupled to component A by covalent bonds or linking functional groups, and then component C is coupled to component B by covalent bonds or linking functional groups to obtain the carrier-drug conjugate; wherein the linking functional groups are selected from one or a combination of amide bonds, ester bonds, thioether bonds, triazole bonds and carbamate bonds.

[0024] According to another aspect of the present invention, a pharmaceutical composition is provided comprising: the carrier drug conjugate of the present invention described above; and optionally pharmaceutically acceptable excipients.

[0025] In some embodiments, the pharmaceutical composition can be formulated into oral preparations such as tablets, capsules, granules, dispersible tablets, effervescent tablets, sustained-release preparations, controlled-release preparations, and enteric-coated preparations.

[0026] According to another aspect of the present invention, the use of the above-described carrier drug conjugate or pharmaceutical composition in the preparation of a medicament for the prevention or treatment of a disease is provided. In some embodiments, the disease is selected from diabetes, obesity, intestinal inflammation, and inflammatory bowel disease. Preferably, the disease is selected from diabetes.

[0027] The above-described carrier-drug conjugates or pharmaceutical compositions of the present invention can be administered orally. The orally administered carrier-drug conjugates or pharmaceutical compositions are non-absorbable in the digestive tract, meaning they do not cross the intestines to enter the systemic circulation. Upon contact with the cell membrane surface, the carrier-drug conjugates or compositions can induce the aggregation of membrane surface receptors towards the conjugate, increasing the local receptor concentration and thereby enhancing the effect of agonist or antagonist signals. The carrier-drug conjugates or compositions can improve affinity for the target site. Specifically, the disease target is located on digestive tract cells, meaning the disease target is on the cell membrane surface of the digestive tract cells with its binding domain facing outwards; specifically, the receptor binder is a drug that can bind to the membrane surface target and stimulate the cell to secrete functional signaling molecules; specifically, the disease is at least one of diabetes, obesity, intestinal inflammation, and inflammatory bowel disease; specifically, the membrane surface target of the digestive tract cell membrane is at least one of bile acid receptor (TGR5), free fatty acid receptor 1 (GPR40), free fatty acid receptor 2 (GPR43), free fatty acid receptor 3 (GPR41), free fatty acid receptor 4 (GPR120), G protein-coupled receptor 119 (GPR119), calcium ion-sensitive receptor (CaSR), and G protein-coupled receptor 35 (GPR35).

[0028] Beneficial effects

[0029] (1) The carrier drug conjugate described in this invention can effectively bind to the target site on the surface of digestive tract cells and is not absorbed by the intestine into the blood circulation.

[0030] (2) The carrier drug conjugate of the present invention also has the ability to amplify receptor agonist or antagonist effects. When the carrier drug conjugate comes into contact with cells, it can promote the accumulation of receptors on the cells at the contact surface, thereby enhancing the effect of agonist or antagonist signals.

[0031] (3) The carrier drug conjugate described in this invention is transported in the digestive tract and does not enter the systemic blood circulation system. It can treat diseases such as diabetes, obesity, intestinal inflammation, and inflammatory bowel disease, while avoiding the toxic side effects that may be caused by a large number of receptor binders entering the body. Attached Figure Description

[0032] To more clearly illustrate the technical solutions in the specific embodiments of this application or the prior art, the drawings used in the description of the specific embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are some embodiments of this application. For those skilled in the art, other drawings can be obtained from these drawings without creative effort.

[0033] Figure 1Hydration kinetics of S500, S1000, R500, and R1000: particle size (A) and surface potential (B).

[0034] Figure 2 Representative carrier-drug conjugate (S500-10% PEG of Example 5) 1000 -DCA, S500-10% PEG of Example 10 2000 -DCA, S500-20% PEG of Example 15 1000 -DCA, S500-20% PEG of Example 19 2000 -DCA, S500-30% PEG of Example 25 1000 -DCA, S500-30% PEG of Example 30 2000 -DCA, S1000-10% PEG of Example 35 1000 -DCA, S1000-10% PEG of Example 40 2000 -DCA, S1000-20% PEG of Example 45 1000 -DCA, S1000-20% PEG of Example 50 2000 -DCA, S1000-30% PEG of Example 55 1000 -DCA, S1000-30% PEG of Example 60 2000 -DCA, R500-10% PEG of Example 65 1000 -DCA, R500-20% PEG of Example 70 1000 -DCA, R500-30% PEG of Example 75 1000 -DCA, R500-10% PEG of Example 80 2000 -DCA, R500-20% PEG of Example 85 2000 -DCA, R500-30% PEG of Example 90 2000 -DCA, R1000-10% PEG of Example 95 1000 -DCA, R1000-20% PEG of Example 96 1000 -DCA, R1000-30% PEG of Example 97 1000 -DCA, R1000-10% PEG of Example 98 2000 -DCA, R1000-20% PEG of Example 99 2000 -DCA, R1000-30% PEG of Example 100 2000 -DCA, R500-20% PEG of Example 105 1000-LCA) and the potentials of their respective carriers and carrier-PEG.

[0035] Figure 3 Representative carrier-drug conjugate (R500-30% PEG of Example 75) 1000 Infrared spectra of PEG(A), DCA(B), R500(C), and R500-30% PEG. 1000 (D) and R500-30% PEG 1000 -Infrared spectrum of DCA(E).

[0036] Figure 4 Representative carrier-drug conjugate (S500-10% PEG of Example 5) 1000 -DCA, S1000-10% PEG of Example 35 1000 -DCA, R500-10% PEG of Example 66 1000 -DCA, R1000-10% PEG of Example 95 1000 Electron micrograph of DCA.

[0037] Figure 5 Gastrointestinal stability of representative carrier-drug conjugates in Example 115. Particle size and potential measurements of the carrier-drug conjugates after incubation for 6 hours in simulated gastric juice (A), simulated small intestinal juice (B), and simulated colonic juice (C).

[0038] Figure 6 Example 116: In vitro nonabsorption validation of representative carrier drug conjugates. Cellular uptake and transcellular transport in Caco-2 cells (A) and E12 cells (B).

[0039] Figure 7 The amount of TGR5 adhering to the surface of a representative carrier drug conjugate in Example 117.

[0040] Figure 8 Dissociation constant (Kd) of the representative carrier drug conjugate and TGR5 protein in Example 118.

[0041] Figure 9 Example 119: In vitro efficacy validation of the representative carrier-drug conjugate. GLP-1 secretion levels in STC-1 cells after 2 hours of incubation with the carrier-drug conjugate.

[0042] Figure 10 In vivo distribution of representative carrier-drug conjugates from Example 120. R500-20% PEG 1000In vivo imaging (A) and semi-quantitative analysis (B) of gastrointestinal transport of DCA and R500 in db / db mice. Dynamic transport in the colon is shown in the upper right.

[0043] Figure 11 : In vivo aggregation validation of representative carrier-drug conjugates from Example 121. R500-20% PEG 1000 -Accumulation status of gold-labeled TGR5 in the colon of mice after DCA administration.

[0044] Figure 12 : In vivo efficacy validation of the representative carrier-drug conjugate in Example 122. R500-20% PEG 1000 Serum GLP-1 levels at 2, 6 and 10 h after DCA administration.

[0045] Figure 13 Pharmacodynamic evaluation of a single dose of the representative carrier drug conjugate in Example 123. Blood glucose changes over time in db / db mice during the oral glucose tolerance test (OGTT) experiment (A) and area under the curve (B).

[0046] Figure 14 Pharmacodynamic evaluation of long-term administration of representative carrier-drug conjugates in Example 124. R500-20% PEG 1000 - The long-term effect of DCA on improving type 2 diabetes in db / db mice. Long-term blood glucose in db / db mice (A), blood glucose changes over time as measured by OGTT on day 35 (B) and area under the curve (C).

[0047] Figure 15 In vivo distribution of representative carrier-drug conjugates from Example 125. Different concentrations of Gd-R500-20% PEG. 1000 -DCA signal intensity (A) and gavage administration of Gd-R500-20% PEG to Bama pigs 1000 - DCA followed by R500-20% PEG 1000 - DCA transport in the gastrointestinal tract (B). Li: liver; Ga: stomach; SB: small intestine; LB: large intestine; Ce: cecum; Co: colon; and Bl: bladder. Scale bar: 10cm.

[0048] Figure 16 Pharmacodynamic evaluation of a single dose of a representative carrier-drug conjugate in Example 126. Blood glucose levels over time in Bama pigs during the OGTT experiment (A) and area under the curve (B).

[0049] Figure 17Long-term pharmacodynamic evaluation of representative carrier-drug conjugates in Example 127. Blood glucose levels (A) and area under the curve (B) of 14-day OGTT in pigs. In (A), red arrows indicate administration, and blue arrows indicate administration of glucose solution.

[0050] Figure 18 : In vivo nonabsorption validation of representative carrier-drug conjugates from Example 128. R500 and R500-20% PEG 1000 - Distribution of DCA in the heart, liver, spleen, lungs and kidneys (A) and fluorescent sections in the liver and kidneys (B).

[0051] Figure 19 In vivo safety validation of the representative carrier drug conjugate in Example 129. In vivo safety validation of the pharmaceutical compositions of the present invention. Relative gallbladder volume (A), bile volume (B), relative liver weight (C), relative liver lipid droplet content (D), ALT level (E), AST level (F), IL-6 level (G), and TNF-α level (H). Figure 20 The diagram illustrates at least the structure contained in the non-absorbable carrier drug conjugate of the present invention. Detailed Implementation

[0052] (I) Introduction

[0053] This invention is partly based on a discovery that a non-absorbable carrier drug conjugate comprising component A with hydration kinetic dimensions between 400 nm and 10,000 nm (particularly between 400 nm and 2,000 nm) and aspect ratios between 1.5 and 15.0 (particularly between 6.0 and 12.0), component B in a molar ratio of said component A between 1,000 and 5,000,000 (particularly between 20,000 and 3,000,000), and component C in a molar ratio of said component A between 100 and 500,000 (particularly between 2,000 and 300,000), exhibits significant therapeutic advantages in treating diseases such as diabetes, obesity, intestinal inflammation, and inflammatory bowel disease. Specifically, as illustrated by the embodiments herein, the non-absorbable carrier drug conjugate provided by this invention advantageously possesses the characteristics of not being absorbed by the intestine into the systemic circulation, enhanced binding to the target site, and promotion of receptor aggregation on the membrane surface. In addition, the non-absorbable carrier drug conjugate of the present invention exhibits good stability in the physiological environment of the digestive tract and can effectively resist the influence of factors such as fluctuating pH, digestive enzymes, flora and intestinal peristalsis in the digestive tract.

[0054] As a non-limiting example, Example 116 shows that 24 embodiments of the non-absorbable carrier drug conjugates of the present invention are non-absorbable compared to previously described small molecule drugs (DCAs). Furthermore, Examples 117 and 118 show that 24 embodiments of the non-absorbable carrier drug conjugates of the present invention exhibit stronger binding affinity to receptor proteins (resulting in stronger activity) compared to previously described small molecule drugs (DCAs).

[0055] As a non-limiting example, Example 119 shows that the 24 non-absorbable carrier drug conjugate embodiments of the present invention have a stronger ability to stimulate target cells to secrete effector protein (GLP-1) compared with previously described small molecule drugs (DCAs) (resulting in stronger drug efficacy).

[0056] As non-limiting examples, Examples 120 and 125 show that a preferred embodiment of the non-absorbable carrier drug conjugate of the present invention can remain in the terminal ileum to the colon for more than 9 hours during gastrointestinal transport. Additionally, Example 121 shows that a non-absorbable carrier drug conjugate embodiment of the present invention can promote the aggregation of intestinal target receptors.

[0057] As a non-limiting example, Example 122 shows that one embodiment of the non-absorbable carrier drug conjugate of the present invention can stimulate the secretion of higher levels of GLP-1 in animals compared with the previously described small molecule drugs (DCAs).

[0058] As non-limiting examples, Examples 123 and 126 show that a non-absorbable carrier drug conjugate embodiment of the present invention exhibits stronger glycemic control in animals compared to previously described small molecule drugs (DCAs) in a single-dose study. Similarly, in long-term treatment, Examples 124 and 127 show that a non-absorbable carrier drug conjugate embodiment of the present invention has a more potent and sustained glycemic lowering effect compared to previously described small molecule drugs (DCAs).

[0059] As a non-limiting example, Example 128 shows that one embodiment of the non-absorbable carrier drug conjugate of the present invention is non-absorbable in mice and does not enter the systemic circulation. Furthermore, after long-term treatment, Example 129 shows that one embodiment of the non-absorbable carrier drug conjugate of the present invention exhibits fewer adverse reactions compared to previously described small molecule drugs (DCAs).

[0060] Importantly, the examples herein illustrate how the presence of a receptor binder and a non-absorbable carrier in a specific molar ratio can enhance the in vivo activity of the non-absorbable carrier drug conjugate of the present invention. For example, the R500-20% PEG described herein... 1000-DCA formulations are particularly advantageous exemplary formulations of the present invention because they exhibit good non-absorption and excellent hypoglycemic effects in vivo, and reduce systemic adverse reactions.

[0061] The non-absorbable carrier drug conjugates of the present invention can be used for a variety of purposes, including targeting and efficiently binding to target receptors in vitro and in vivo. Therefore, the present invention provides a method for treating diseases or functional impairments in subjects requiring treatment, which is achieved by contacting said subjects with the non-absorbable carrier drug conjugates described herein.

[0062] Different exemplary embodiments of the non-absorbable carrier drug conjugates of the present invention are described in further detail below.

[0063] (II) Definition

[0064] 1. Carrier-drug conjugates (CDCs): These are a class of targeted drugs formed by the coupling of a carrier and a receptor binder through specific linkers. They are a type of drug that is non-absorbable, highly targeted, and has relatively low toxicity.

[0065] 2. Non-absorbability in vivo: Non-absorbability refers to the property of a drug with an absorption fraction (Fabs) < 85%. Fabs refers to the proportion of drug dose absorbed from the gastrointestinal tract into the systemic circulation after oral administration. The calculation formula is as follows:

[0066]

[0067] 3. Cellular nonabsorbability: This refers to the phenomenon where substances cannot be absorbed or taken up by cells. This characteristic is usually determined by the physicochemical properties of the substance itself, the barrier function of the cell membrane, or the biological mechanisms of the cell. Assay methods include (DouTY, Wang J et al., International Journal of Biological Macromolecules, 2019): Caco-2 cells are seeded in 24-well plates and cultured to a suitable density. A reagent is added and incubated for 2 hours. The reagent is discarded, and the cells are washed with PBS. Cell lysates are collected and fluorescence is measured. The fluorescence intensity of non-absorbable reagents is 1% or less of that of other reagents. Alternatively, E12 cells are seeded in 24-well Transwell chambers and cultured until a dense cell monolayer forms. A reagent is then added to the chamber, and 200 μL samples are taken from the base medium at 2, 4, and 8 hours to measure fluorescence intensity. The fluorescence intensity of non-absorbable reagents is close to 0.

[0068] 4. Digestive Tract Physiological Environment: The digestive tract physiological environment refers to the biological and physicochemical characteristics of each part of the digestive tract from the mouth to the anus, as well as the internal environmental conditions related to functions such as digestion, absorption, and excretion. It includes the anatomical structure, pH value, enzyme activity, microbial community, transport mechanisms, and other characteristics of each segment of the digestive tract.

[0069] 5. TGR5 agonists: G protein-coupled bile acid receptor agonists

[0070] 6. GPR40 agonists: G protein-coupled receptor 40 agonists

[0071] 7. GPR120 agonists: G protein-coupled receptor 120 agonists

[0072] 8. GPR41 agonists: G protein-coupled receptor 41 agonists

[0073] 9. GPR43 agonists: G protein-coupled receptor 43 agonists

[0074] 10. GPR119 agonists: G protein-coupled receptor 119 agonists

[0075] 11. CaSR agonists: calcium ion-sensitive receptor agonists

[0076] 12. GPR35 agonists: G protein-coupled receptor 35 agonists

[0077] 13. Hydration kinetic dimensions: These refer to the effective dimensions of the carrier and its surrounding hydrated molecular layer in an aqueous solution. They emphasize the interaction between the carrier particle surface and water molecules and can be measured using techniques such as dynamic light scattering.

[0078] 14. Receptor binders: These are molecules that can specifically bind to and interact with receptors on the cell surface or inside the cell. They can regulate receptor function by binding to receptors and are commonly used in areas such as targeted drug therapy.

[0079] 15. Aspect Ratio: This is a parameter describing the shape characteristics of a particle. It is usually used to measure the size ratio of a particle in two perpendicular directions. Specifically, the aspect ratio is the ratio between the longest dimension (length) and the shortest dimension (width) of a particle.

[0080] 16. Dissociation constant: The dissociation constant Kd is the equilibrium dissociation constant between the antibody and its antigen, i.e., k off / k on The ratio of Kd to affinity. Kd is inversely proportional to affinity; the lower the Kd value, the higher the affinity of the antibody.

[0081] 17. Silica: An inorganic non-metallic material with good chemical stability and biocompatibility. It is commonly used in drug delivery systems as a mesoporous carrier material, and efficient drug loading and release can be controlled by adjusting its pore size, specific surface area, and surface functionalization.

[0082] 18. Graphene oxide: An oxidized derivative of graphene, containing abundant hydroxyl, carboxyl, and epoxy groups, possessing good water solubility, functionalization modification ability, and drug loading capacity, enabling targeted and responsive release in drug delivery.

[0083] 19. Carbon quantum dots: A type of carbon-based fluorescent material with nanoscale dimensions, possessing excellent optical properties, biocompatibility, and low toxicity, which can be used for imaging and drug delivery carriers in drug delivery systems.

[0084] 20. Hydroxyapatite: A calcium phosphate material with good biocompatibility with bone tissue, chemical formula Ca. 10 (PO4)6(OH)2. Due to its biodegradability and excellent drug loading capacity, it is widely used in bone repair and sustained-release drug delivery systems.

[0085] 21. Titanium dioxide: An inorganic metal oxide with high chemical stability and good biocompatibility. Its nanostructures, due to their photoresponsive properties and surface modification capabilities, can be applied to drug carriers and photosensitized drug delivery systems.

[0086] 22. Metal-organic frameworks: a class of porous crystalline materials composed of metal ions or clusters and organic ligands through coordination bonds. They have high specific surface area, tunable pore size and good drug loading capacity, and are widely used in controlled release drug delivery systems.

[0087] 23. Covalent organic framework: A porous crystalline organic polymer assembled by covalent bonds, with high chemical stability and tunable pore structure, used in drug delivery to load drugs and achieve efficient, targeted release.

[0088] 24. Poly(lactic acid-glycolic acid): A biodegradable copolymer composed of lactic acid and glycolic acid monomers. Its degradation rate can be controlled by adjusting the ratio of lactic acid to glycolic acid. It is widely used in drug delivery systems such as microspheres and nanoparticles.

[0089] 25. Carbon nanotubes: Hollow nanostructures formed by rolling up graphene sheets. They come in single-walled and multi-walled forms and have high specific surface area, good electrical conductivity and mechanical strength. They can be functionalized through surface modification for efficient drug loading and targeted delivery.

[0090] 26. Krebs Ringer Buffer (KR Buffer): KR buffer can mimic the physiological environment of the intestine. Its components are 135mM NaCl, 3.6mM KCl, 1.2mM MgCl2, 0.5mM NaH2PO4, 5mM NaHCO3, 1.5mM CaCl2 and 10mM 4-hydroxyethylpiperazine ethanesulfonic acid (HEPES).

[0091] (III) Description of the Implementation Plan

[0092] This invention provides a class of non-absorbable carrier drug conjugates, a method for preparing the non-absorbable carrier drug conjugates, and applications of the non-absorbable carrier drug conjugates (e.g., for the treatment of diseases or functional disorders).

[0093] In one aspect, the present invention provides a carrier-drug conjugate system comprising: component A, wherein the material of component A is selected from inorganic and organic materials; preferably, it is selected from one or a combination of silica, graphene oxide, carbon quantum dots, carbon nanotubes, hydroxyapatite, titanium dioxide, metal-organic frameworks, covalent organic frameworks, and poly(lactic-co-glycolic acid); more preferably, it is selected from one or a combination of silica, poly(lactic-co-glycolic acid), and carbon nanotubes; most preferably, the material of component A is silica. The hydration kinetic size of component A is greater than or equal to 400 nm to 10000 nm; more preferably, the hydration kinetic size is between 400 nm and 2000 nm. The aspect ratio of component A is ≥1.25; preferably, the aspect ratio is ≥1.5; more preferably, the aspect ratio is between 1.5 and 15.0; most preferably, the aspect ratio is between 6.0 and 12.0. Component B, after being attached to the surface of component A, increases its hydration kinetic size by 5 to 60 nm. The material of component B is selected from hydrophobic or hydrophilic polymers; preferably, from hydrophilic polymers; more preferably, from polyethylene glycol and its derivatives containing 5 or more -CH2-CH2-O repeating units; most preferably, from polyethylene glycol derivatives, characterized by containing 16 to 24 -CH2-CH2-O repeating units and having one end of the molecule modified to an amino group. The molar ratio of component B to component A is between 1,000 and 5,000,000; preferably, the molar ratio of component B to component A is between 5,000 and 4,000,000; more preferably, the molar ratio of component B to component A is between 10,000 and 4,000,000; most preferably, the molar ratio of component B to component A is between 20,000 and 3,000,000. Component C is derived from the material of component C, which is selected from one or a combination of TGR5 agonists, GPR40 agonists, GPR120 agonists, GPR41 agonists, GPR43 agonists, GPR119 agonists, CaSR agonists, and GPR35 agonists. Preferably, the material of component C is selected from TGR5 agonists and GPR35 agonists; more preferably, the material of component C is selected from bile acid compounds and their analogues, INT-777, zapustol, pamoic acid, lodusamide, and YE120; even more preferably, the material of component C is selected from cholic acid, deoxycholic acid, chenodeoxycholic acid, ursodeoxycholic acid, lithocholic acid, pamoic acid, etc.; most preferably, the material of component C is selected from deoxycholic acid and pamoic acid.The molar ratio of component C to component A is between 100 and 500,000; preferably, the molar ratio of component C to component A is between 300 and 400,000; more preferably, the molar ratio of component C to component A is between 500 and 400,000; and most preferably, the molar ratio of component C to component A is between 2,000 and 300,000.

[0094] Component A

[0095] In some embodiments, component A is non-absorbable and stable in the physiological environment of the digestive tract, and is intended for administration via the digestive route, capable of binding to specific targets and not absorbed into systemic circulation. It also remains stable in the digestive tract environment. In some other embodiments, component A is substantially non-toxic to mammals such as humans. In some embodiments, component A comprises inorganic materials, such as graphene oxide, carbon quantum dots, carbon nanotubes, hydroxyapatite, titanium dioxide, or a mixture of graphene oxide and carbon quantum dots. In other embodiments, component A comprises organic materials, such as metal-organic frameworks, covalent organic frameworks, poly(lactic-co-glycolic acid), or mixtures thereof. In the most preferred embodiment, component A is silicon dioxide.

[0096] Component A

[0097] In some embodiments of the non-absorbable carrier drug conjugate of the present invention, the hydration kinetic size of component A can be between 400 nm and 500 nm. In other embodiments, the hydration kinetic size of component A can be between 500 nm and 1100 nm. In other embodiments, the hydration kinetic size of component A can be between 1100 nm and 2000 nm. In other embodiments, the hydration kinetic size of component A can be between 2000 nm and 10,000 nm. In still other embodiments, the hydration kinetic size of component A can be 466, 1045, or 1500 nm (or any portion thereof or a range thereof). Typically, the aspect ratio of component A is between 6.0 and 12.0. In some cases, the aspect ratio of component A can range from 1.25 to 1.5, 1.5 to 6, 12 to 15, or >15. Component A of the present invention exhibits good stability and non-absorbability in the physiological environment of the digestive tract.

[0098] Component B material

[0099] Component B, as a linker, shows broad application potential in drug delivery systems. Its design can utilize hydrophobic or hydrophilic polymers, especially hydrophilic polyethylene glycol (PEG) and its derivatives, particularly PEG derivatives containing 16–24 repeating -CH2-CH2-O units with an amino group modified at one end of the molecule. The chain length of Component B can be controlled within the range of 5–60 nm. This design not only improves the hydration kinetics of the drug delivery carrier but also optimizes its stability and biocompatibility.

[0100] Specifically, the linkers of component B can be connected to component A via amide bonds, ester bonds, thioether bonds, triazole bonds, or carbamate bonds, and support different concentrations (e.g., 10%, 20%, or 30%) of PEG1000 or PEG2000. In the most preferred embodiment, component B is selected from a derivative of PEG1000, characterized in that one end of the molecule is modified to be amino.

[0101] Content of Component B

[0102] In some embodiments of the non-absorbable carrier drug conjugate of the present invention, the molar ratio of component B to component A must be within a specific range to ensure that components B and C have suitable surface densities. Density χ is determined by PEG modification. peg (mg / g), Avogadro's constant (NA), density ρ of component A measured by the bulk density method A (g / cm 3 ) and the calculated volume v of component A A (cm 3 The molar ratio (n) of component B to component A is calculated using the following formula:

[0103] n = χ peg ×NA×ρ A ×v A

[0104] The molar ratio calculated according to the above formula can be 1000–10,000, 10,000–20,000, 3,000,000–4,000,000, or 4,000,000–5,000,000. In another embodiment, the molar ratio of component B to component A is 146797, 362846, 464966, 1174383, 2902769, 3719731, 21423, 44473, or 61391 (or any part thereof or range thereof). Component B of the present invention can optimize the hydration characteristics of the delivery carrier and has a certain degree of flexibility and suitable length, which is beneficial to the interaction between component C and the target.

[0105] Component C material

[0106] In some other embodiments, component C is substantially non-toxic to mammals such as humans. In some embodiments, component C comprises a free fatty acid receptor agonist, such as a GPR40 agonist, GPR43 agonist, GPR120 agonist, and GPR41 agonist. In other embodiments, component C comprises a G protein-coupled receptor and a calcium ion-sensitive receptor, such as a GPR119 agonist, GPR35 agonist, and CaSR agonist. In other embodiments, component C comprises a TGR5 agonist, such as INT-777, deoxycholic acid, chenodeoxycholic acid, ursodeoxycholic acid, lithocholic acid, etc. In the most preferred embodiment, component C is deoxycholic acid (DCA).

[0107] Content of component C

[0108] In some embodiments of the non-absorbable carrier drug conjugate of the present invention, the molar ratio of component C to component A must be within a specific range to ensure that the drug has a suitable surface density, and the density χ is modified by DCA. DCA (mg / g), Avogadro's constant (NA), density ρ of component A measured by the bulk density method A (g / cm 3 ) and the calculated volume v of component A A (cm 3 The molar ratio (n) of component C to component A is calculated using the following formula:

[0109] n = χ DCA ×NA×ρ A ×v A

[0110] The molar ratio is calculated using the above formula, and the molar ratio of component C to component A can be 10. 3 ~10 4 10 4 ~10 5 10 5 ~10 6 Or 10 6 ~10 7 In other embodiments, the molar ratio of component C to component A can be 10. 3 ~3×10 3 3×10 3 ~5×10 3 Or 5×10 3 ~10 4In another embodiment, the molar ratio of component C to component A is 137736, 278016, 334107, 1101891, 2224134, 2672863, 15179, 38429, or 45982 (or any part thereof or a range thereof). Component C of the present invention has a suitable density distribution, which can improve the affinity with the target and promote the aggregation of receptors on the membrane surface.

[0111] In one embodiment of the non-absorbable carrier drug conjugate of the present invention, it comprises a non-absorbable mesoporous silica carrier R500 and PEG. 1000 When the carrier drug conjugate composed of (20%) and DCA (20%) was administered orally in a db / db diabetic mouse model, the non-absorbable carrier drug conjugate was distributed only in the digestive tract and was not absorbed into the blood or systemic circulation, thus exhibiting non-absorbability. This avoided adverse reactions such as gallbladder toxicity, liver toxicity, and inflammation. At the same time, the mice's anti-glycemic ability was improved, and blood glucose levels continued to decrease to a stable level, which is a significant advantage compared to the traditional small molecule agonist DCA.

[0112] Example

[0113] The present invention will be described in detail below by way of examples. However, the examples provided herein are for illustrative purposes only and are not intended to limit the invention.

[0114] Unless otherwise specified, the experimental methods used in the following examples are conventional methods.

[0115] Unless otherwise specified, all materials and reagents used in the following examples are commercially available.

[0116] Table 1 Experimental Materials and Suppliers

[0117]

[0118]

[0119]

[0120] Note: Other common reagents such as NaOH, KH2PO4, anhydrous ethanol, glycerol, methanol, ammonia, methanol and concentrated hydrochloric acid were purchased from Sinopharm Chemical Reagent Co., Ltd.

[0121] Table 2. Experimental Instruments and Manufacturers

[0122]

[0123]

[0124] Experimental cells: Caco-2 cells and HT29-MTX-E12 (E12) cells were purchased from the cell bank of the Shanghai Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences, and cultured in DMEM complete medium. HEK293T cells were purchased from the cell bank of the Shanghai Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences, and cultured in DMEM complete medium during the experiment. STC-1 cells were donated by the research group of Li Jingya at the Shanghai Institute of Materia Medica, Chinese Academy of Sciences, and cultured in DMEM complete medium supplemented with 1% L-glutamine during the experiment. NCI-H716 cells were purchased from the cell bank of the Shanghai Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences, and cultured in RPMI 1640 complete medium. All cell culture media contained 10% (v / v) fetal bovine serum and 1% penicillin and streptomycin, and were cultured in a constant temperature incubator at 37°C and 5% CO2.

[0125] Laboratory animals: Male 7-week-old db / db mice were purchased from Jiangsu Jicui Pharmaceutical Biotechnology Co., Ltd. and housed in an SPF-grade animal facility. 7-week-old male C57BL / 6 (C57) mice were purchased from the Animal Experiment Center of Shanghai Institute of Materia Medica and housed in an SPF-grade animal facility. Male Bama pigs weighing 20±2 kg were purchased from Shanghai Jiagan Biotechnology Co., Ltd. All animals were fed a normal diet and kept in a constant-temperature environment with a 12-hour light / dark cycle. All animals underwent at least one week of acclimatization before the experiments. The experimental protocols were approved by the International Association for the Use and Management of Laboratory Animals (IACUC) (IMM experiments: IACUC: 2021-08-GY-61; pig experiments: IACUC: PZSHUTCM2211070010). All experimental procedures were strictly performed in accordance with IACUC regulations.

[0126] Example 1: Preparation of non-absorbent mesoporous silica support S500

[0127] S1: Preparation of seed suspension. 25 mL of tetraethoxysilane (TEOS) was dispersed in 45 mL of methanol and added dropwise to a mixture of 210 mL of methanol and 45 mL of ammonia. The mixture was reacted at 500 rpm for 5 h at room temperature.

[0128] S2: Formation of micelle structure. Take 7.6 mL of the seed suspension prepared in S1, mix it with 154 mL of methanol and 78 mL of ammonia water, and stir at 500 rpm at room temperature.

[0129] S3: Formation of the support precursor. A mixture of 1.56 mL TEOS and 98 mL methanol was added dropwise to the micelle solution formed in S2, and the mixture was reacted at 500 rpm for 4 h at room temperature.

[0130] S4: Centrifuge the solution in S3 at 10,000 rpm, wash the precipitate with anhydrous ethanol, resuspend it in 20 mL of ethanol, add 3 mL of concentrated hydrochloric acid, and stir at 10,000 rpm at 65 °C for more than 8 hours.

[0131] S5: Deacidification. Centrifuge the carrier solution prepared in S4 at 10,000 rpm, wash twice with ultrapure water and anhydrous ethanol respectively, and finally resuspend in anhydrous ethanol for storage.

[0132] This embodiment successfully prepared a mesoporous silica support S500 with non-absorbent properties through the above five steps. The particle size and potential of S500 are as follows: Figure 1 As shown, its hydration kinetic particle size is approximately 500 nm, and its potential is below -20 mV.

[0133] Example 2: Preparation of non-absorbent mesoporous silica support S1000

[0134] S1: Preparation of seed suspension. 25 mL of LTEOS was dispersed in 45 mL of methanol and added dropwise to a mixture of 210 mL of methanol and 45 mL of ammonia. The mixture was reacted at 500 rpm for 5 h at room temperature.

[0135] S2: Formation of micelle structure. Take 43.5 mL of the seed suspension prepared in S1 and add it to a mixture of 47 mL of methanol and 29 mL of ammonia. Stir at 500 rpm at room temperature.

[0136] S3: Formation of the support precursor. A mixture of 0.78 mL TEOS and 49 mL methanol was added dropwise to the micelle solution formed in S2, and the mixture was reacted at 500 rpm for 24 h at room temperature.

[0137] S4: Centrifuge the solution in S3 at 8,000 rpm, wash the precipitate with anhydrous ethanol, resuspend it in 20 mL of ethanol, add 3 mL of concentrated hydrochloric acid, and stir at 8,000 rpm at 65 °C for more than 8 hours.

[0138] S5: Deacidification. Centrifuge the carrier solution prepared in S4 at 8,000 rpm, wash twice with ultrapure water and anhydrous ethanol respectively, and finally resuspend in anhydrous ethanol for storage.

[0139] This embodiment successfully prepared a mesoporous silica support S1000 with non-absorbent properties through the above five steps. The particle size and potential of S1000 are as follows: Figure 1 As shown, its hydration kinetic particle size is approximately 1000 nm, and its potential is below -20 mV.

[0140] Example 3: Preparation of non-absorbent mesoporous silica support R500

[0141] S1: Formation of micelle structure. 0.55 g cetyltrimethylammonium bromide (CTAB) was dissolved in 54 mL of ultrapure water, and then 0.87 mL of ammonia water was added. The mixture was stirred at 500 rpm for 1 h at room temperature.

[0142] S2: Formation of the carrier precursor. Add 0.52 mL of LTEOS dropwise to the micelle solution formed in S1 and continue stirring for 3 hours.

[0143] S3: Centrifuge the solution in S2 at 10,000 rpm, wash the precipitate with anhydrous ethanol, resuspend it in 20 mL of ethanol, add 3 mL of concentrated hydrochloric acid, and stir at 10,000 rpm at 65 °C for more than 8 hours.

[0144] S4: Deacidification. Centrifuge the carrier solution prepared in S3 at 10,000 rpm, wash twice with ultrapure water and anhydrous ethanol respectively, and finally resuspend in anhydrous ethanol for storage.

[0145] This embodiment successfully prepared a mesoporous silica support R500 with non-absorbent properties through the above four steps. The particle size and potential of R500 are as follows: Figure 1 As shown, its hydration kinetic particle size is approximately 500 nm, and its potential is below -20 mV.

[0146] Example 4: Preparation of non-absorbable poly(lactic-co-glycolic acid) carrier R1000

[0147] S1: Preparation of poly(lactic-co-glycolic acid) microspheres. 50 mg of poly(lactic-co-glycolic acid) (PLGA-COOH) was dissolved in 1.5 mL of chloroform and mixed with a 1% polyvinyl alcohol (PVA, w / v) solution. The mixture was then homogenized in an ice-water bath at 2000 rpm for 5 min using a homogenizer. Subsequently, the chloroform was removed by rotary evaporation to obtain poly(lactic-co-glycolic acid) microspheres.

[0148] S2: Preparation of PVA film-forming solution. Dissolve 8g of PVA in 40mL of deionized water at 90℃ and 500rpm until completely dissolved. Add 0.8mL of glycerol and continue stirring for 10min. Allow to cool naturally to room temperature to obtain the PVA film-forming solution.

[0149] S3: PVA film preparation. Take 2.5–5 mL of PVA film-forming solution and pour it onto a film-forming plate (25 cm²). 2 In the mixture, shake to disperse the film-forming solution evenly, and add 5 mg / mL of poly(lactic acid-glycolic acid) microsphere solution. Shake on a shaker at room temperature for 15 min, and dry in an oven at 37°C for more than 15 h to obtain a PVA film with a thickness of 150-300 μm.

[0150] S4: PVA film stretching. The PVA film was cut into 3cm × 8cm pieces, bathed in an oil bath at 70℃ for 10 minutes, and then stretched from 3cm × 8cm to approximately 2.5cm × 10cm using a custom-designed device at a speed of 0.5mm / s. It was then kept in cold mineral oil for 5 minutes to lower the temperature, and then washed with ether to remove residual oil.

[0151] S5: PVA removal. The stretched PVA film was dissolved in deionized water and centrifuged at 2000 rpm for 20 min to remove gel-like deposits; the supernatant was centrifuged at 15500 g for 30 min, resuspended in deionized water, and the particles were washed at least five times to remove PVA, yielding poly(lactic-co-glycolic acid) microrods.

[0152] This embodiment successfully prepared a non-absorbent poly(lactic-co-glycolic acid) support R1000 through the above five steps. The particle size and potential of R1000 are as follows: Figure 1 As shown, its hydration kinetic particle size is approximately 1000 nm, and its potential is below -20 mV.

[0153] Example 5: 10% PEG with amide bonds 1000 -DCA-modified S500 mesoporous silica (S500-10% PEG) 1000 -DCA)

[0154] S1: Mesoporous silica dispersion. 500 mg of the non-absorbent mesoporous silica support S500 prepared in Example 1 was dispersed in a mixture of 340 mL water and 200 mL ethanol, and the pH was adjusted to 4.0 with dilute hydrochloric acid.

[0155] S2: 10% PEG 1000 -NH2 modification. 10% silane-PEG was added to S1. 1000 The NH2 ethanol solution was added, and then ethanol solution was added to make the ethanol:water volume ratio of the reaction system 1:2. The reaction was carried out at 65℃ and 800rpm for 24h. The carrier was washed by centrifugation and resuspended in anhydrous ethanol to obtain 10% PEG. 1000- NH2-modified S500 solution.

[0156] S3: Deoxycholic acid (DCA) carboxyl activation. Dissolve 109.8 mg DCA in 11.0 mL N,N-dimethylformamide (DMF). Then add 21 mg N-hydroxysuccinimide (NHS) and 56 mg 1-ethyl-(-dimethylaminopropyl)carbodiimide (EDC) sequentially, activate at room temperature for 15 min, and adjust pH to 5.8.

[0157] S4: DCA-modified mesoporous silica. The activation solution of S3 was added to 50 mL of an ethanol solution of S2 (10 mg / mL). The reaction was carried out at room temperature and 500 rpm in the dark for 24 h. The mixture was centrifuged and washed to obtain 10% DCA-modified S500 mesoporous silica. The results are as follows... Figure 4 As shown, S500-10% PEG 1000 -DCA appears spherical under an electron microscope, with a diameter of approximately 500 nm. S500-10% PEG 1000 and S500-10% PEG 1000 - The particle size and potential of DCA are as follows Figure 2 As shown, the surface potential gradually approaches electroneutrality from electronegativity, and then becomes electronegative again after DCA grafting. This may be because the carboxyl groups of DCA bind to the amino groups on the PEG surface, weakening the charge shielding effect of PEG itself. Characterization of the potential demonstrates the successful modification with DCA.

[0158] Example 6: 10% PEG with ester bonds 1000 DCA-modified S500 mesoporous silicon

[0159] S1: Mesoporous silica dispersion. 500 mg of the non-absorbent mesoporous silica support S500 prepared in Example 1 was dispersed in a mixture of 340 mL water and 200 mL ethanol, and the pH was adjusted to 4.0 with dilute hydrochloric acid.

[0160] S2: 10% PEG 1000 -OH modification. Add 10% silane-PEG to S1. 1000 -OH ethanol solution was added, and then ethanol solution was added to make the ethanol:water volume ratio of the reaction system 1:2. The reaction was carried out at 65℃ and 800rpm for 24h. The carrier was washed by centrifugation and resuspended in anhydrous ethanol to obtain 10% PEG. 1000- OH-modified S500 solution.

[0161] S3: DCA carboxyl activation. 100 mg DCA was dissolved in 5 mL of LDM under a dry nitrogen atmosphere. 125 mg DCC was added, and the mixture was stirred at room temperature for 1 hour to form the activated intermediate O-acylurea.

[0162] S4: DCA-modified mesoporous silica. The intermediate from S3 was added to 5 mg of DPTS, followed by the addition of 10% PEG from S2. 1000 The modified mesoporous silica was stirred and reacted for 12-24 hours. The mixture was centrifuged and washed to obtain 10% DCA-modified S500 mesoporous silica.

[0163] Example 7: S500 mesoporous silica modified with 10% PEG1000-DCA and linked by thioether bonds.

[0164] S1: Mesoporous silica dispersion. 500 mg of the non-absorbent mesoporous silica support S500 prepared in Example 1 was dispersed in a mixture of 340 mL water and 200 mL ethanol, and the pH was adjusted to 4.0 with dilute hydrochloric acid.

[0165] S2: 10% PEG 1000 -Mal modification. Add 10% silane-PEG to S1. 1000 The ethanol solution was first prepared, and then ethanol was added to make the ethanol:water volume ratio of the reaction system 1:2. The reaction was carried out at 65°C and 800 rpm for 24 h. The carrier was washed by centrifugation and resuspended in DMF to obtain 10% PEG. 1000 -Mal-modified S500 solution.

[0166] S3: DCA-modified mesoporous silica. 100 mg of thiolized DCA was dissolved in 10 mL of LDM under a dry nitrogen atmosphere. 10% PEG from S2 was then added. 1000 The modified mesoporous silica was stirred and reacted for 12-24 hours. The mixture was centrifuged and washed to obtain 10% DCA-modified S500 mesoporous silica.

[0167] Example 8: 10% PEG with triazole linkage 1000 DCA-modified S500 mesoporous silicon

[0168] S1: Mesoporous silica dispersion. 500 mg of the non-absorbent mesoporous silica support S500 prepared in Example 1 was dispersed in a mixture of 340 mL water and 200 mL ethanol, and the pH was adjusted to 4.0 with dilute hydrochloric acid.

[0169] S2: 10% PEG 1000 -N3 modification. 10% silane-PEG was added to S1. 1000 -N3 ethanol solution was added, and then ethanol solution was added to make the ethanol:water volume ratio of the reaction system 1:2. The reaction was carried out at 65℃ and 800rpm for 24h. The carrier was washed by centrifugation and resuspended in DMF to obtain 10% PEG. 1000 -N3 modified ratio of S500 solution.

[0170] S3: DCA-modified mesoporous silica. Under a dry nitrogen atmosphere, 100 mg of alkynylated DCA, 5 mg of CuSO4·5H2O, and 10 mg of sodium ascorbate were dissolved in 5 mL of DMF. 10% PEG from S2 was then added. 1000 The modified mesoporous silica was reacted with stirring at room temperature for 12 h. The mixture was centrifuged and washed to obtain 10% DCA-modified S500 mesoporous silica.

[0171] Example 9: 10% PEG with carbamate bonds 1000 DCA-modified S500 mesoporous silicon

[0172] S1: Mesoporous silica dispersion. 500 mg of the non-absorbent mesoporous silica support S500 prepared in Example 1 was dispersed in a mixture of 340 mL water and 200 mL ethanol, and the pH was adjusted to 4.0 with dilute hydrochloric acid.

[0173] S2: 10% PEG 1000 -NCO modification. Add 10% silane-PEG to S1. 1000 -NCO ethanol solution was added, and then ethanol solution was added to make the ethanol:water volume ratio of the reaction system 1:2. The reaction was carried out at 65℃ and 800rpm for 24h. The carrier was washed by centrifugation and resuspended in DMF to obtain 10% PEG. 1000 -NCO modified S500 solution.

[0174] S3: DCA-modified mesoporous silica. 100 mg of isocyanated DCA was dissolved in 5 mL of LDM under a dry nitrogen atmosphere. 10 mg of DMAP was added as a catalyst, along with 10% PEG from S2. 1000 The modified mesoporous silica was reacted with stirring at room temperature for 6-12 hours. The mixture was centrifuged and washed to obtain 10% DCA-modified S500 mesoporous silica.

[0175] Example 10: 10% PEG with amide bonds 2000 -DCA-modified S500 mesoporous silica (S500-10% PEG) 2000 -DCA)

[0176] S1: Mesoporous silica dispersion. 500 mg of the non-absorbent mesoporous silica support S500 prepared in Example 1 was dispersed in a mixture of 340 mL water and 200 mL ethanol, and the pH was adjusted to 4.0 with dilute hydrochloric acid.

[0177] S2: 10% PEG 2000 -NH2 modification. 10% silane-PEG was added to S1. 2000 The NH2 ethanol solution was added, and then ethanol solution was added to make the ethanol:water volume ratio of the reaction system 1:2. The reaction was carried out at 65℃ and 800rpm for 24h. The carrier was washed by centrifugation and resuspended in anhydrous ethanol to obtain 10% PEG. 2000 S500 solution with -NH2 modification ratio.

[0178] S3: DCA carboxyl activation. Dissolve 109.8 mg DCA in 11.0 mL DMF. Then add 21 mg NHS and 56 mg EDC sequentially, activate at room temperature for 15 min, and adjust pH to 5.8.

[0179] S4: DCA-modified mesoporous silica. The activation solution of S3 was added to 50 mL of ethanol solution (10 mg / mL). The reaction was carried out at room temperature and 500 rpm in the dark for 24 h. The mixture was centrifuged and washed to obtain 10% DCA-modified S500 mesoporous silica. S500-10% PEG 2000 and S500-10% PEG 2000 - The particle size and potential of DCA are as follows Figure 2 As shown, the surface potential gradually approaches electroneutrality from electronegativity, and then becomes electronegative again after DCA grafting. This may be because the carboxyl groups of DCA bind to the amino groups on the PEG surface, weakening the charge shielding effect of PEG itself. Characterization of the potential demonstrates the successful modification with DCA.

[0180] Example 11: 10% PEG with ester bonds 2000 DCA-modified S500 mesoporous silicon

[0181] S1: Mesoporous silica dispersion. 500 mg of the non-absorbent mesoporous silica support S500 prepared in Example 1 was dispersed in a mixture of 340 mL water and 200 mL ethanol, and the pH was adjusted to 4.0 with dilute hydrochloric acid.

[0182] S2: 10% PEG 2000 -OH modification. Add 10% silane-PEG to S1. 2000 -OH ethanol solution was added, and then ethanol solution was added to make the ethanol:water volume ratio of the reaction system 1:2. The reaction was carried out at 65℃ and 800rpm for 24h. The carrier was washed by centrifugation and resuspended in anhydrous ethanol to obtain 10% PEG. 2000 S500 solution with -OH modification ratio.

[0183] S3: DCA carboxyl activation. 100 mg DCA was dissolved in 5 mL of LDM under a dry nitrogen atmosphere. 125 mg DCC was added, and the mixture was stirred at room temperature for 1 hour to form the activated intermediate O-acylurea.

[0184] S4: DCA-modified mesoporous silica. 5 mg of DPTS was added to the intermediate in S3, followed by the addition of 10% PEG from S2. 2000 The modified mesoporous silica was stirred and reacted for 12-24 hours. The mixture was centrifuged and washed to obtain 10% DCA-modified S500 mesoporous silica.

[0185] Example 12: 10% PEG with thioether linkage 2000 DCA-modified S500 mesoporous silicon

[0186] S1: Mesoporous silica dispersion. 500 mg of the non-absorbent mesoporous silica support S500 prepared in Example 1 was dispersed in a mixture of 340 mL water and 200 mL ethanol, and the pH was adjusted to 4.0 with dilute hydrochloric acid.

[0187] S2: 10% PEG 2000 -Mal modification. Add 10% silane-PEG to S1. 2000 The ethanol solution was first prepared, and then ethanol was added to make the ethanol:water volume ratio of the reaction system 1:2. The reaction was carried out at 65°C and 800 rpm for 24 h. The carrier was washed by centrifugation and resuspended in DMF to obtain 10% PEG. 2000 -Mal modified S500 solution.

[0188] S3: DCA-modified mesoporous silica. 100 mg of thiolized DCA was dissolved in 10 mL of LDM under a dry nitrogen atmosphere. 10% PEG from S2 was then added. 2000 The modified mesoporous silica was stirred and reacted for 12-24 hours. The mixture was centrifuged and washed to obtain 10% DCA-modified S500 mesoporous silica.

[0189] Example 13: 10% PEG with triazole linkage 2000 DCA-modified S500 mesoporous silicon

[0190] S1: Mesoporous silica dispersion. 500 mg of the non-absorbent mesoporous silica support S500 prepared in Example 1 was dispersed in a mixture of 340 mL water and 200 mL ethanol, and the pH was adjusted to 4.0 with dilute hydrochloric acid.

[0191] S2: 10% PEG 2000 -N3 modification. 10% silane-PEG was added to S1. 2000 -N3 ethanol solution was added, and then ethanol solution was added to make the ethanol:water volume ratio of the reaction system 1:2. The reaction was carried out at 65℃ and 800rpm for 24h. The carrier was washed by centrifugation and resuspended in DMF to obtain 10% PEG. 2000 -N3 modified S500 solution.

[0192] S3: DCA-modified mesoporous silica. Under a dry nitrogen atmosphere, 100 mg of alkynylated DCA, 5 mg of CuSO4·5H2O, and 10 mg of sodium ascorbate were dissolved in 5 mL of DMF. 10% PEG from S2 was then added. 2000The modified mesoporous silica was reacted with stirring at room temperature for 12 h. The mixture was centrifuged and washed to obtain 10% DCA-modified S500 mesoporous silica.

[0193] Example 14: 10% PEG with carbamate linkages 2000 DCA-modified S500 mesoporous silicon

[0194] S1: Mesoporous silica dispersion. 500 mg of the non-absorbent mesoporous silica support S500 prepared in Example 1 was dispersed in a mixture of 340 mL water and 200 mL ethanol, and the pH was adjusted to 4.0 with dilute hydrochloric acid.

[0195] S2: 10% PEG 2000 -NCO modification. Add 10% silane-PEG to S1. 2000 -NCO ethanol solution was added, and then ethanol solution was added to make the ethanol:water volume ratio of the reaction system 1:2. The reaction was carried out at 65℃ and 800rpm for 24h. The carrier was washed by centrifugation and resuspended in DMF to obtain 10% PEG. 2000 -NCO modified S500 solution.

[0196] S3: DCA-modified mesoporous silica. 100 mg of isocyanated DCA was dissolved in 5 mL of LDM under a dry nitrogen atmosphere. 10 mg of DMAP was added as a catalyst, along with 10% PEG from S2. 2000 The modified mesoporous silica was reacted with stirring at room temperature for 6-12 hours. The mixture was centrifuged and washed to obtain 10% DCA-modified S500 mesoporous silica.

[0197] Example 15: 20% PEG with amide bonds 1000 -DCA-modified S500 mesoporous silicon (S500-20% PEG) 1000 -DCA)

[0198] S1: Mesoporous silica dispersion. 500 mg of the non-absorbent mesoporous silica support S500 prepared in Example 1 was dispersed in a mixture of 340 mL water and 200 mL ethanol, and the pH was adjusted to 4.0 with dilute hydrochloric acid.

[0199] S2: 20% PEG 1000 -NH2 modification. 20% silane-PEG was added to S1. 1000 The NH2 ethanol solution was added, and then ethanol solution was added to make the ethanol:water volume ratio of the reaction system 1:2. The reaction was carried out at 65℃ and 800rpm for 24h. The carrier was washed by centrifugation and resuspended in anhydrous ethanol to obtain 20% PEG. 1000- NH2-modified S500 solution.

[0200] S3: DCA carboxyl activation. Dissolve 109.8 mg DCA in 11.0 mL DMF. Then add 21 mg NHS and 56 mg EDC sequentially, activate at room temperature for 15 min, and adjust pH to 5.8.

[0201] S4: DCA-modified mesoporous silica. The activation solution of S3 was added to 50 mL of ethanol solution (10 mg / mL). The reaction was carried out at room temperature and 500 rpm in the dark for 24 h. The mixture was centrifuged and washed to obtain 20% DCA-modified S500 mesoporous silica. S500-20% PEG 1000 and S500-20% PEG 1000 - The particle size and potential of DCA are as follows Figure 2 As shown, the surface potential gradually approaches electroneutrality from electronegativity, and then becomes electronegative again after DCA grafting. This may be because the carboxyl groups of DCA bind to the amino groups on the PEG surface, weakening the charge shielding effect of PEG itself. Characterization of the potential demonstrates the successful modification with DCA.

[0202] Example 16: 20% PEG with ester bonds 1000 DCA-modified S500 mesoporous silicon

[0203] S1: Mesoporous silica dispersion. 500 mg of the non-absorbent mesoporous silica support S500 prepared in Example 1 was dispersed in a mixture of 340 mL water and 200 mL ethanol, and the pH was adjusted to 4.0 with dilute hydrochloric acid.

[0204] S2: 20% PEG 1000 -OH modification. Add 20% silane-PEG to S1. 1000 -OH ethanol solution was added, and then ethanol solution was added to make the ethanol:water volume ratio of the reaction system 1:2. The reaction was carried out at 65℃ and 800rpm for 24h. The carrier was washed by centrifugation and resuspended in anhydrous ethanol to obtain 20% PEG. 1000- OH-modified S500 solution.

[0205] S3: DCA carboxyl activation. Under a dry nitrogen atmosphere, 100 mg DCA was dissolved in 5 mL DMF. 125 mg DCC was added, and the mixture was stirred at room temperature for 1 h to form the activated intermediate O-acylurea.

[0206] S4: DCA-modified mesoporous silica. The intermediate from S3 was added to 5 mg of DPTS, followed by the addition of 20% PEG from S2. 1000 The modified mesoporous silica was stirred and reacted for 12-24 hours. The mixture was centrifuged and washed to obtain 20% DCA-modified S500 mesoporous silica.

[0207] Example 17: 20% PEG1000-DCA modified S500 mesoporous silica with thioether bonds

[0208] S1: Mesoporous silica dispersion. 500 mg of the non-absorbent mesoporous silica support S500 prepared in Example 1 was dispersed in a mixture of 340 mL water and 200 mL ethanol, and the pH was adjusted to 4.0 with dilute hydrochloric acid.

[0209] S2: 20% PEG 1000 -Mal modification. Add 20% silane-PEG to S1. 1000 The ethanol solution was first prepared, and then ethanol was added to make the ethanol:water volume ratio of the reaction system 1:2. The reaction was carried out at 65°C and 800 rpm for 24 h. The carrier was washed by centrifugation and resuspended in DMF to obtain 20% PEG. 1000 -Mal-modified S500 solution.

[0210] S3: DCA-modified mesoporous silica. 100 mg of thiolized DCA was dissolved in 10 mL of LDM under a dry nitrogen atmosphere. 20% PEG from S2 was then added. 1000 The modified mesoporous silica was stirred and reacted for 12-24 hours. The mixture was centrifuged and washed to obtain 20% DCA-modified S500 mesoporous silica.

[0211] Example 18: 20% PEG with triazole linkage 1000 DCA-modified S500 mesoporous silicon

[0212] S1: Mesoporous silica dispersion. 500 mg of the non-absorbent mesoporous silica support S500 prepared in Example 1 was dispersed in a mixture of 340 mL water and 200 mL ethanol, and the pH was adjusted to 4.0 with dilute hydrochloric acid.

[0213] S2: 20% PEG 1000 -N3 modification. 20% silane-PEG was added to S1. 1000 -N3 ethanol solution was added, and then ethanol solution was added to make the ethanol:water volume ratio of the reaction system 1:2. The reaction was carried out at 65℃ and 800rpm for 24h. The carrier was washed by centrifugation and resuspended in DMF to obtain 20% PEG. 1000 -N3 modified ratio of S500 solution.

[0214] S3: DCA-modified mesoporous silica. Under a dry nitrogen atmosphere, 100 mg of alkynylated DCA, 5 mg of CuSO4·5H2O, and 10 mg of sodium ascorbate were dissolved in 5 mL of DMF. 20% of the PEG from S2 was then added. 1000The modified mesoporous silica was reacted with stirring at room temperature for 12 h. The mixture was centrifuged and washed to obtain 20% DCA-modified S500 mesoporous silica.

[0215] Example 19: 20% PEG with carbamate bonds 1000 -DCA-modified S500 mesoporous silica (S500-20% PEG) 2000 -DCA)

[0216] S1: Mesoporous silica dispersion. 500 mg of the non-absorbent mesoporous silica support S500 prepared in Example 1 was dispersed in a mixture of 340 mL water and 200 mL ethanol, and the pH was adjusted to 4.0 with dilute hydrochloric acid.

[0217] S2: 20% PEG 1000 -NCO modification. Add 20% silane-PEG to S1. 1000 -NCO ethanol solution was added, and then ethanol solution was added to make the ethanol:water volume ratio of the reaction system 1:2. The reaction was carried out at 65℃ and 800rpm for 24h. The carrier was washed by centrifugation and resuspended in DMF to obtain 20% PEG. 1000 -NCO modified S500 solution.

[0218] S3: DCA-modified mesoporous silica. 100 mg of isocyanated DCA was dissolved in 5 mL of LDM under a dry nitrogen atmosphere. 10 mg of DMAP was added as a catalyst, along with 20% PEG from S2. 1000 The modified mesoporous silica was reacted with stirring at room temperature for 6-12 h. The mixture was centrifuged and washed to obtain 20% DCA-modified S500 mesoporous silica.

[0219] Example 20: 20% PEG with amide bonds 2000 DCA-modified S500 mesoporous silicon

[0220] S1: Mesoporous silica dispersion. 500 mg of the non-absorbent mesoporous silica support S500 prepared in Example 1 was dispersed in a mixture of 340 mL water and 200 mL ethanol, and the pH was adjusted to 4.0 with dilute hydrochloric acid.

[0221] S2: 20% PEG 2000 -NH2 modification. 20% silane-PEG was added to S1. 2000 The NH2 ethanol solution was added, and then ethanol solution was added to make the ethanol:water volume ratio of the reaction system 1:2. The reaction was carried out at 65℃ and 800rpm for 24h. The carrier was washed by centrifugation and resuspended in anhydrous ethanol to obtain 20% PEG. 2000 S500 solution with -NH2 modification ratio.

[0222] S3: DCA carboxyl activation. Dissolve 109.8 mg DCA in 11.0 mL DMF. Then add 21 mg NHS and 56 mg EDC sequentially, activate at room temperature for 15 min, and adjust pH to 5.8.

[0223] S4: DCA-modified mesoporous silica. The activation solution of S3 was added to 50 mL of ethanol solution (10 mg / mL). The reaction was carried out at room temperature and 500 rpm in the dark for 24 h. The mixture was centrifuged and washed to obtain 20% DCA-modified S500 mesoporous silica. S500-20% PEG 2000 and S500-20% PEG 2000 - The particle size and potential of DCA are as follows Figure 2 As shown, the surface potential gradually approaches electroneutrality from electronegativity, and then becomes electronegative again after DCA grafting. This may be because the carboxyl groups of DCA bind to the amino groups on the PEG surface, weakening the charge shielding effect of PEG itself. Characterization of the potential demonstrates the successful modification with DCA.

[0224] Example 21: 20% PEG with ester bonds 2000 DCA-modified S500 mesoporous silicon

[0225] S1: Mesoporous silica dispersion. 500 mg of the non-absorbent mesoporous silica support S500 prepared in Example 1 was dispersed in a mixture of 340 mL water and 200 mL ethanol, and the pH was adjusted to 4.0 with dilute hydrochloric acid.

[0226] S2: 20% PEG 2000 -OH modification. Add 20% silane-PEG to S1. 2000 -OH ethanol solution was added, and then ethanol solution was added to make the ethanol:water volume ratio of the reaction system 1:2. The reaction was carried out at 65℃ and 800rpm for 24h. The carrier was washed by centrifugation and resuspended in anhydrous ethanol to obtain 20% PEG. 2000 S500 solution with -OH modification ratio.

[0227] S3: DCA carboxyl activation. Under a dry nitrogen atmosphere, 100 mg DCA was dissolved in 5 mL DMF. 125 mg DCC was added, and the mixture was stirred at room temperature for 1 h to form the activated intermediate O-acylurea.

[0228] S4: DCA-modified mesoporous silica. 5 mg of DPTS was added to the intermediate in S3, followed by the addition of 20% PEG from S2. 2000 The modified mesoporous silica was stirred and reacted for 12-24 hours. The mixture was centrifuged and washed to obtain 20% DCA-modified S500 mesoporous silica.

[0229] Example 22: 20% PEG with thioether linkages 2000 DCA-modified S500 mesoporous silicon

[0230] S1: Mesoporous silica dispersion. 500 mg of the non-absorbent mesoporous silica support S500 prepared in Example 1 was dispersed in a mixture of 340 mL water and 200 mL ethanol, and the pH was adjusted to 4.0 with dilute hydrochloric acid.

[0231] S2: 20% PEG 2000 -Mal modification. Add 20% silane-PEG to S1. 2000 The ethanol solution was first prepared, and then ethanol was added to make the ethanol:water volume ratio of the reaction system 1:2. The reaction was carried out at 65°C and 800 rpm for 24 h. The carrier was washed by centrifugation and resuspended in DMF to obtain 20% PEG. 2000 -Mal modified S500 solution.

[0232] S3: DCA-modified mesoporous silica. 100 mg of thiolized DCA was dissolved in 10 mL of LDM under a dry nitrogen atmosphere. 20% PEG from S2 was then added. 2000 The modified mesoporous silica was stirred and reacted for 12-24 hours. The mixture was centrifuged and washed to obtain 20% DCA-modified S500 mesoporous silica.

[0233] Example 23: 20% PEG with triazole linkage 2000 DCA-modified S500 mesoporous silicon

[0234] S1: Mesoporous silica dispersion. 500 mg of the non-absorbent mesoporous silica support S500 prepared in Example 1 was dispersed in a mixture of 340 mL water and 200 mL ethanol, and the pH was adjusted to 4.0 with dilute hydrochloric acid.

[0235] S2: 20% PEG 2000 -N3 modification. 20% silane-PEG was added to S1. 2000 -N3 ethanol solution was added, and then ethanol solution was added to make the ethanol:water volume ratio of the reaction system 1:2. The reaction was carried out at 65℃ and 800rpm for 24h. The carrier was washed by centrifugation and resuspended in DMF to obtain 20% PEG. 2000 -N3 modified S500 solution.

[0236] S3: DCA-modified mesoporous silica. Under a dry nitrogen atmosphere, 100 mg of alkynylated DCA, 5 mg of CuSO4·5H2O, and 10 mg of sodium ascorbate were dissolved in 5 mL of DMF. 20% of the PEG from S2 was then added. 2000The modified mesoporous silica was reacted with stirring at room temperature for 12 h. The mixture was centrifuged and washed to obtain 20% DCA-modified S500 mesoporous silica.

[0237] Example 24: 20% PEG with carbamate bonds 2000 DCA-modified S500 mesoporous silicon

[0238] S1: Mesoporous silica dispersion. 500 mg of the non-absorbent mesoporous silica support S500 prepared in Example 1 was dispersed in a mixture of 340 mL water and 200 mL ethanol, and the pH was adjusted to 4.0 with dilute hydrochloric acid.

[0239] S2: 20% PEG 2000 -NCO modification. Add 20% silane-PEG to S1. 2000 -NCO ethanol solution was added, and then ethanol solution was added to make the ethanol:water volume ratio of the reaction system 1:2. The reaction was carried out at 65℃ and 800rpm for 24h. The carrier was washed by centrifugation and resuspended in DMF to obtain 20% PEG. 2000 -NCO modified S500 solution.

[0240] S3: DCA-modified mesoporous silica. 100 mg of isocyanated DCA was dissolved in 5 mL of LDM under a dry nitrogen atmosphere. 10 mg of DMAP was added as a catalyst, along with 20% PEG from S2. 2000 The modified mesoporous silica was reacted with stirring at room temperature for 6-12 h. The mixture was centrifuged and washed to obtain 20% DCA-modified S500 mesoporous silica.

[0241] Example 25: 30% PEG with amide bonds 1000 -DCA-modified S500 mesoporous silica (S500-30% PEG) 1000 -DCA)

[0242] S1: Mesoporous silica dispersion. 500 mg of the non-absorbent mesoporous silica support S500 prepared in Example 1 was dispersed in a mixture of 340 mL water and 200 mL ethanol, and the pH was adjusted to 4.0 with dilute hydrochloric acid.

[0243] S2: 30% PEG 1000 -NH2 modification. 30% silane-PEG was added to S1. 1000 -NH2 ethanol solution was added, and then ethanol solution was added to make the ethanol:water volume ratio of the reaction system 1:2. The reaction was carried out at 65℃ and 800rpm for 24h. The carrier was washed by centrifugation and resuspended in anhydrous ethanol to obtain 30% PEG. 1000- NH2-modified S500 solution.

[0244] S3: DCA carboxyl activation. Dissolve 109.8 mg DCA in 11.0 mL DMF. Then add 21 mg NHS and 56 mg EDC sequentially, activate at room temperature for 15 min, and adjust pH to 5.8.

[0245] S4: DCA-modified mesoporous silica. The activation solution of S3 was added to 50 mL of an ethanol solution of S2 (10 mg / mL). The reaction was carried out at room temperature and 500 rpm in the dark for 24 h. The mixture was centrifuged and washed to obtain 30% DCA-modified S500 mesoporous silica. S500-30% PEG 1000 and S500-30% PEG 1000 - The particle size and potential of DCA are as follows Figure 2 As shown, the surface potential gradually approaches electroneutrality from electronegativity, and then becomes electronegative again after DCA grafting. This may be because the carboxyl groups of DCA bind to the amino groups on the PEG surface, weakening the charge shielding effect of PEG itself. Characterization of the potential demonstrates the successful modification with DCA.

[0246] Example 26: 30% PEG with ester bonds 1000 DCA-modified S500 mesoporous silicon

[0247] S1: Mesoporous silica dispersion. 500 mg of the non-absorbent mesoporous silica support S500 prepared in Example 1 was dispersed in a mixture of 340 mL water and 200 mL ethanol, and the pH was adjusted to 4.0 with dilute hydrochloric acid.

[0248] S2: 30% PEG 1000 -OH modification. Add 30% silane-PEG to S1. 1000 -OH ethanol solution was added, and then ethanol solution was added to make the ethanol:water volume ratio of the reaction system 1:2. The reaction was carried out at 65℃ and 800rpm for 24h. The carrier was washed by centrifugation and resuspended in anhydrous ethanol to obtain 30% PEG. 1000- OH-modified S500 solution.

[0249] S3: DCA carboxyl activation. 100 mg DCA was dissolved in 5 mL of LDM under a dry nitrogen atmosphere. 125 mg DCC was added, and the mixture was stirred at room temperature for 1 hour to form the activated intermediate O-acylurea.

[0250] S4: DCA-modified mesoporous silica. The intermediate from S3 was added to 5 mg of DPTS, followed by the addition of 30% PEG from S2. 1000 The modified mesoporous silica was stirred and reacted for 12-24 hours. The mixture was centrifuged and washed to obtain 30% DCA-modified S500 mesoporous silica.

[0251] Example 27: 30% PEG1000-DCA modified S500 mesoporous silica with thioether bonds

[0252] S1: Mesoporous silica dispersion. 500 mg of the non-absorbent mesoporous silica support S500 prepared in Example 1 was dispersed in a mixture of 340 mL water and 200 mL ethanol, and the pH was adjusted to 4.0 with dilute hydrochloric acid.

[0253] S2: 30% PEG 1000 -Mal modification. Add 30% silane-PEG to S1. 1000 The ethanol solution was first prepared, and then ethanol was added to make the ethanol:water volume ratio of the reaction system 1:2. The reaction was carried out at 65°C and 800 rpm for 24 h. The carrier was washed by centrifugation and resuspended in DMF to obtain 30% PEG. 1000 -Mal-modified S500 solution.

[0254] S3: DCA-modified mesoporous silica. 100 mg of thiolized DCA was dissolved in 10 mL of LDM under a dry nitrogen atmosphere. 30% of the PEG from S2 was then added. 1000 The modified mesoporous silica was stirred and reacted for 12-24 hours. The mixture was centrifuged and washed to obtain 30% DCA-modified S500 mesoporous silica.

[0255] Example 28: 30% PEG with triazole linkage 1000 DCA-modified S500 mesoporous silicon

[0256] S1: Mesoporous silica dispersion. 500 mg of the non-absorbent mesoporous silica support S500 prepared in Example 1 was dispersed in a mixture of 340 mL water and 200 mL ethanol, and the pH was adjusted to 4.0 with dilute hydrochloric acid.

[0257] S2: 30% PEG 1000 -N3 modification. 30% silane-PEG was added to S1. 1000 -N3 ethanol solution was added, and then ethanol solution was added to make the ethanol:water volume ratio of the reaction system 1:2. The reaction was carried out at 65℃ and 800rpm for 24h. The carrier was washed by centrifugation and resuspended in DMF to obtain 30% PEG. 1000 -N3 modified ratio of S500 solution.

[0258] S3: DCA-modified mesoporous silica. Under a dry nitrogen atmosphere, 100 mg of alkynylated DCA, 5 mg of CuSO4·5H2O, and 10 mg of sodium ascorbate were dissolved in 5 mL of DMF. 30% of the PEG from S2 was then added. 1000The modified mesoporous silica was reacted with stirring at room temperature for 12 h. The mixture was centrifuged and washed to obtain 30% DCA-modified S500 mesoporous silica.

[0259] Example 29: 30% PEG with carbamate bonds 1000 DCA-modified S500 mesoporous silicon

[0260] S1: Mesoporous silica dispersion. 500 mg of the non-absorbent mesoporous silica support S500 prepared in Example 1 was dispersed in a mixture of 340 mL water and 200 mL ethanol, and the pH was adjusted to 4.0 with dilute hydrochloric acid.

[0261] S2: 30% PEG 1000 -NCO modification. Add 30% silane-PEG to S1. 1000 -NCO ethanol solution was added, and then ethanol solution was added to make the ethanol:water volume ratio of the reaction system 1:2. The reaction was carried out at 65℃ and 800rpm for 24h. The carrier was washed by centrifugation and resuspended in DMF to obtain 30% PEG. 1000 -NCO modified S500 solution.

[0262] S3: DCA-modified mesoporous silica. 100 mg of isocyanated DCA was dissolved in 5 mL of LDM under a dry nitrogen atmosphere. 10 mg of DMAP was added as a catalyst, along with 30% PEG from S2. 1000 The modified mesoporous silica was reacted with stirring at room temperature for 6-12 h. The mixture was centrifuged and washed to obtain 30% DCA-modified S500 mesoporous silica.

[0263] Example 30: 30% PEG with amide bonds 2000 -DCA-modified S500 mesoporous silica (S500-30% PEG) 2000 -DCA)

[0264] S1: Mesoporous silica dispersion. 500 mg of the non-absorbent mesoporous silica support S500 prepared in Example 1 was dispersed in a mixture of 340 mL water and 200 mL ethanol, and the pH was adjusted to 4.0 with dilute hydrochloric acid.

[0265] S2: 30% PEG 2000 -NH2 modification. 30% silane-PEG was added to S1. 2000 -NH2 ethanol solution was added, and then ethanol solution was added to make the ethanol:water volume ratio of the reaction system 1:2. The reaction was carried out at 65℃ and 800rpm for 24h. The carrier was washed by centrifugation and resuspended in anhydrous ethanol to obtain 30% PEG. 2000 S500 solution with -NH2 modification ratio.

[0266] S3: DCA carboxyl activation. Dissolve 109.8 mg DCA in 11.0 mL DMF. Then add 21 mg NHS and 56 mg EDC sequentially, activate at room temperature for 15 min, and adjust pH to 5.8.

[0267] S4: DCA-modified mesoporous silica. The activation solution of S3 was added to 50 mL of ethanol solution (10 mg / mL). The reaction was carried out at room temperature and 500 rpm in the dark for 24 h. The mixture was centrifuged and washed to obtain 30% DCA-modified S500 mesoporous silica. S500-30% PEG 2000 and S500-30% PEG 2000 - The particle size and potential of DCA are as follows Figure 2 As shown, the surface potential gradually approaches electroneutrality from electronegativity, and then becomes electronegative again after DCA grafting. This may be because the carboxyl groups of DCA bind to the amino groups on the PEG surface, weakening the charge shielding effect of PEG itself. Characterization of the potential demonstrates the successful modification with DCA.

[0268] Example 31: 30% PEG with ester bonds 2000 DCA-modified S500 mesoporous silicon

[0269] S1: Mesoporous silica dispersion. 500 mg of the non-absorbent mesoporous silica support S500 prepared in Example 1 was dispersed in a mixture of 340 mL water and 200 mL ethanol, and the pH was adjusted to 4.0 with dilute hydrochloric acid.

[0270] S2: 30% PEG 2000 -OH modification. Add 30% silane-PEG to S1. 2000 -OH ethanol solution was added, and then ethanol solution was added to make the ethanol:water volume ratio of the reaction system 1:2. The reaction was carried out at 65℃ and 800rpm for 24h. The carrier was washed by centrifugation and resuspended in anhydrous ethanol to obtain 30% PEG. 2000 S500 solution with -OH modification ratio.

[0271] S3: DCA carboxyl activation. 100 mg DCA was dissolved in 5 mL of LDM under a dry nitrogen atmosphere. 125 mg DCC was added, and the mixture was stirred at room temperature for 1 hour to form the activated intermediate O-acylurea.

[0272] S4: DCA-modified mesoporous silica. 5 mg of DPTS was added to the intermediate in S3, followed by the addition of 30% PEG from S2. 2000 The modified mesoporous silica was stirred and reacted for 12-24 hours. The mixture was centrifuged and washed to obtain 30% DCA-modified S500 mesoporous silica.

[0273] Example 32: 30% PEG with thioether linkages 2000 DCA-modified S500 mesoporous silicon

[0274] S1: Mesoporous silica dispersion. 500 mg of the non-absorbent mesoporous silica support S500 prepared in Example 1 was dispersed in a mixture of 340 mL water and 200 mL ethanol, and the pH was adjusted to 4.0 with dilute hydrochloric acid.

[0275] S2: 30% PEG 2000 -Mal modification. Add 30% silane-PEG to S1. 2000 The ethanol solution was first prepared, and then ethanol was added to make the ethanol:water volume ratio of the reaction system 1:2. The reaction was carried out at 65°C and 800 rpm for 24 h. The carrier was washed by centrifugation and resuspended in DMF to obtain 30% PEG. 2000 -Mal modified S500 solution.

[0276] S3: DCA-modified mesoporous silica. 100 mg of thiolized DCA was dissolved in 10 mL of LDM under a dry nitrogen atmosphere. 30% of the PEG from S2 was then added. 2000 The modified mesoporous silica was stirred and reacted for 12-24 hours. The mixture was centrifuged and washed to obtain 30% DCA-modified S500 mesoporous silica.

[0277] Example 33: 30% PEG with triazole linkage 2000 DCA-modified S500 mesoporous silicon

[0278] S1: Mesoporous silica dispersion. 500 mg of the non-absorbent mesoporous silica support S500 prepared in Example 1 was dispersed in a mixture of 340 mL water and 200 mL ethanol, and the pH was adjusted to 4.0 with dilute hydrochloric acid.

[0279] S2: 30% PEG 2000 -N3 modification. 30% silane-PEG was added to S1. 2000 -N3 ethanol solution was added, and then ethanol solution was added to make the ethanol:water volume ratio of the reaction system 1:2. The reaction was carried out at 65℃ and 800rpm for 24h. The carrier was washed by centrifugation and resuspended in DMF to obtain 30% PEG. 2000 -N3 modified S500 solution.

[0280] S3: DCA-modified mesoporous silica. Under a dry nitrogen atmosphere, 100 mg of alkynylated DCA, 5 mg of CuSO4·5H2O, and 10 mg of sodium ascorbate were dissolved in 5 mL of DMF. 30% of the PEG from S2 was then added. 2000The modified mesoporous silica was reacted with stirring at room temperature for 12 h. The mixture was centrifuged and washed to obtain 30% DCA-modified S500 mesoporous silica.

[0281] Example 34: 30% PEG with carbamate bonds 2000 DCA-modified S500 mesoporous silicon

[0282] S1: Mesoporous silica dispersion. 500 mg of the non-absorbent mesoporous silica support S500 prepared in Example 1 was dispersed in a mixture of 340 mL water and 200 mL ethanol, and the pH was adjusted to 4.0 with dilute hydrochloric acid.

[0283] S2: 30% PEG 2000 -NCO modification. Add 30% silane-PEG to S1. 2000 -NCO ethanol solution was added, and then ethanol solution was added to make the ethanol:water volume ratio of the reaction system 1:2. The reaction was carried out at 65℃ and 800rpm for 24h. The carrier was washed by centrifugation and resuspended in DMF to obtain 30% PEG. 2000 -NCO modified S500 solution.

[0284] S3: DCA-modified mesoporous silica. 100 mg of isocyanated DCA was dissolved in 5 mL of LDM under a dry nitrogen atmosphere. 10 mg of DMAP was added as a catalyst, along with 30% PEG from S2. 2000 The modified mesoporous silica was reacted with stirring at room temperature for 6-12 h. The mixture was centrifuged and washed to obtain 30% DCA-modified S500 mesoporous silica.

[0285] Example 35: 10% PEG with amide bonds 1000 -DCA-modified S1000 mesoporous silica (S1000-10% PEG) 1000 -DCA)

[0286] S1: Mesoporous silica dispersion. 500 mg of the non-absorbent mesoporous silica carrier S1000 prepared in Example 2 was dispersed in a mixture of 340 mL water and 200 mL ethanol, and the pH was adjusted to 4.0 with dilute hydrochloric acid.

[0287] S2: 10% PEG 1000 -NH2 modification. 10% silane-PEG was added to S1. 1000 The NH2 ethanol solution was added, and then ethanol solution was added to make the ethanol:water volume ratio of the reaction system 1:2. The reaction was carried out at 65℃ and 800rpm for 24h. The carrier was washed by centrifugation and resuspended in anhydrous ethanol to obtain 10% PEG. 1000- NH2-modified S1000 solution.

[0288] S3: DCA carboxyl activation. Dissolve 109.8 mg DCA in 11.0 mL DMF. Then add 21 mg NHS and 56 mg EDC sequentially, activate at room temperature for 15 min, and adjust pH to 5.8.

[0289] S4: DCA-modified mesoporous silica. The activation solution of S3 was added to 50 mL of an ethanol solution of S2 (10 mg / mL). The reaction was carried out at room temperature and 500 rpm in the dark for 24 h. The mixture was centrifuged and washed to obtain 10% DCA-modified S1000 mesoporous silica. The results are as follows... Figure 4 As shown, S1000-10% PEG 1000 -DCA appears spherical under an electron microscope, with a diameter of approximately 1000 nm. S1000-10% PEG 1000 and S1000-10% PEG 1000 - The particle size and potential of DCA are as follows Figure 2 As shown, the surface potential gradually approaches electroneutrality from electronegativity, and then becomes electronegative again after DCA grafting. This may be because the carboxyl groups of DCA bind to the amino groups on the PEG surface, weakening the charge shielding effect of PEG itself. Characterization of the potential demonstrates the successful modification with DCA.

[0290] Example 36: 10% PEG with ester bonds 1000 DCA-modified S1000 mesoporous silicon

[0291] S1: Mesoporous silica dispersion. 500 mg of the non-absorbent mesoporous silica carrier S1000 prepared in Example 2 was dispersed in a mixture of 340 mL water and 200 mL ethanol, and the pH was adjusted to 4.0 with dilute hydrochloric acid.

[0292] S2: 10% PEG 1000 -OH modification. Add 10% silane-PEG to S1. 1000 -OH ethanol solution was added, and then ethanol solution was added to make the ethanol:water volume ratio of the reaction system 1:2. The reaction was carried out at 65℃ and 800rpm for 24h. The carrier was washed by centrifugation and resuspended in anhydrous ethanol to obtain 10% PEG. 1000- OH-modified S1000 solution.

[0293] S3: DCA carboxyl activation. Under a dry nitrogen atmosphere, 100 mg DCA was dissolved in 5 mL DMF. 125 mg DCC was added, and the mixture was stirred at room temperature for 1 h to form the activated intermediate O-acylurea.

[0294] S4: DCA-modified mesoporous silica. The intermediate from S3 was added to 5 mg of DPTS, followed by the addition of 10% PEG from S2.1000 The modified mesoporous silica was stirred and reacted for 12-24 hours. The mixture was centrifuged and washed to obtain 10% DCA-modified S1000 mesoporous silica.

[0295] Example 37: 10% PEG1000-DCA modified S1000 mesoporous silica with thioether linkages

[0296] S1: Mesoporous silica dispersion. 500 mg of the non-absorbent mesoporous silica carrier S1000 prepared in Example 2 was dispersed in a mixture of 340 mL water and 200 mL ethanol, and the pH was adjusted to 4.0 with dilute hydrochloric acid.

[0297] S2: 10% PEG 1000 -Mal modification. Add 10% silane-PEG to S1. 1000 The ethanol solution was first prepared, and then ethanol was added to make the ethanol:water volume ratio of the reaction system 1:2. The reaction was carried out at 65°C and 800 rpm for 24 h. The carrier was washed by centrifugation and resuspended in DMF to obtain 10% PEG. 1000 -Mal-modified S1000 solution.

[0298] S3: DCA-modified mesoporous silica. 100 mg of thiolized DCA was dissolved in 10 mL of LDM under a dry nitrogen atmosphere. 10% PEG from S2 was then added. 1000 The modified mesoporous silica was stirred and reacted for 12-24 hours. The mixture was centrifuged and washed to obtain 10% DCA-modified S1000 mesoporous silica.

[0299] Example 38: 10% PEG with triazole linkage 1000 DCA-modified S1000 mesoporous silicon

[0300] S1: Mesoporous silica dispersion. 500 mg of the non-absorbent mesoporous silica carrier S1000 prepared in Example 2 was dispersed in a mixture of 340 mL water and 200 mL ethanol, and the pH was adjusted to 4.0 with dilute hydrochloric acid.

[0301] S2: 10% PEG 1000 -N3 modification. 10% silane-PEG was added to S1. 1000 -N3 ethanol solution was added, and then ethanol solution was added to make the ethanol:water volume ratio of the reaction system 1:2. The reaction was carried out at 65℃ and 800rpm for 24h. The carrier was washed by centrifugation and resuspended in DMF to obtain 10% PEG. 1000 -N3 modified ratio of S1000 solution.

[0302] S3: DCA-modified mesoporous silica. Under a dry nitrogen atmosphere, 100 mg of alkynylated DCA, 5 mg of CuSO4·5H2O, and 10 mg of sodium ascorbate were dissolved in 5 mL of DMF. 10% PEG from S2 was then added. 1000 The modified mesoporous silica was reacted with stirring at room temperature for 12 h. The mixture was centrifuged and washed to obtain 10% DCA-modified S1000 mesoporous silica.

[0303] Example 39: 10% PEG with carbamate bonds 1000 DCA-modified S1000 mesoporous silicon

[0304] S1: Mesoporous silica dispersion. 500 mg of the non-absorbent mesoporous silica carrier S1000 prepared in Example 2 was dispersed in a mixture of 340 mL water and 200 mL ethanol, and the pH was adjusted to 4.0 with dilute hydrochloric acid.

[0305] S2: 10% PEG 1000 -NCO modification. Add 10% silane-PEG to S1. 1000 -NCO ethanol solution was added, and then ethanol solution was added to make the ethanol:water volume ratio of the reaction system 1:2. The reaction was carried out at 65℃ and 800rpm for 24h. The carrier was washed by centrifugation and resuspended in DMF to obtain 10% PEG. 1000 -NCO modified S1000 solution.

[0306] S3: DCA-modified mesoporous silica. 100 mg of isocyanated DCA was dissolved in 5 mL of LDM under a dry nitrogen atmosphere. 10 mg of DMAP was added as a catalyst, along with 10% PEG from S2. 1000 The modified mesoporous silica was reacted with stirring at room temperature for 6-12 hours. The mixture was centrifuged and washed to obtain 10% DCA-modified S1000 mesoporous silica.

[0307] Example 40: 10% PEG with amide bonds 2000 -DCA-modified S1000 mesoporous silica (S1000-10% PEG) 2000 -DCA)

[0308] S1: Mesoporous silica dispersion. 500 mg of the non-absorbent mesoporous silica carrier S1000 prepared in Example 2 was dispersed in a mixture of 340 mL water and 200 mL ethanol, and the pH was adjusted to 4.0 with dilute hydrochloric acid.

[0309] S2: 10% PEG 2000 -NH2 modification. 10% silane-PEG was added to S1. 2000The NH2 ethanol solution was added, and then ethanol solution was added to make the ethanol:water volume ratio of the reaction system 1:2. The reaction was carried out at 65℃ and 800rpm for 24h. The carrier was washed by centrifugation and resuspended in anhydrous ethanol to obtain 10% PEG. 2000 S1000 solution with -NH2 modification ratio.

[0310] S3: DCA carboxyl activation. Dissolve 109.8 mg DCA in 11.0 mL DMF. Then add 21 mg NHS and 56 mg EDC sequentially, activate at room temperature for 15 min, and adjust pH to 5.8.

[0311] S4: DCA-modified mesoporous silica. The activation solution of S3 was added to 50 mL of ethanol solution (10 mg / mL). The reaction was carried out at room temperature and 500 rpm in the dark for 24 h. The mixture was centrifuged and washed to obtain 10% DCA-modified S1000 mesoporous silica. S1000-10% PEG 2000 and S1000-10% PEG 2000 - The particle size and potential of DCA are as follows Figure 2 As shown, the surface potential gradually approaches electroneutrality from electronegativity, and then becomes electronegative again after DCA grafting. This may be because the carboxyl groups of DCA bind to the amino groups on the PEG surface, weakening the charge shielding effect of PEG itself. Characterization of the potential demonstrates the successful modification with DCA.

[0312] Example 41: 10% PEG with ester bonds 2000 DCA-modified S1000 mesoporous silicon

[0313] S1: Mesoporous silica dispersion. 500 mg of the non-absorbent mesoporous silica carrier S1000 prepared in Example 2 was dispersed in a mixture of 340 mL water and 200 mL ethanol, and the pH was adjusted to 4.0 with dilute hydrochloric acid.

[0314] S2: 10% PEG 2000 -OH modification. Add 10% silane-PEG to S1. 2000 -OH ethanol solution was added, and then ethanol solution was added to make the ethanol:water volume ratio of the reaction system 1:2. The reaction was carried out at 65℃ and 800rpm for 24h. The carrier was washed by centrifugation and resuspended in anhydrous ethanol to obtain 10% PEG. 2000 S1000 solution with -OH modification ratio.

[0315] S3: DCA carboxyl activation. Under a dry nitrogen atmosphere, 100 mg DCA was dissolved in 5 mL DMF. 125 mg DCC was added, and the mixture was stirred at room temperature for 1 h to form the activated intermediate O-acylurea.

[0316] S4: DCA-modified mesoporous silica. 5 mg of DPTS was added to the intermediate in S3, followed by the addition of 10% PEG from S2. 2000 The modified mesoporous silica was stirred and reacted for 12-24 hours. The mixture was centrifuged and washed to obtain 10% DCA-modified S1000 mesoporous silica.

[0317] Example 42: 10% PEG with thioether linkage 2000 DCA-modified S1000 mesoporous silicon

[0318] S1: Mesoporous silica dispersion. 500 mg of the non-absorbent mesoporous silica carrier S1000 prepared in Example 2 was dispersed in a mixture of 340 mL water and 200 mL ethanol, and the pH was adjusted to 4.0 with dilute hydrochloric acid.

[0319] S2: 10% PEG 2000 -Mal modification. Add 10% silane-PEG to S1. 2000 The ethanol solution was first prepared, and then ethanol was added to make the ethanol:water volume ratio of the reaction system 1:2. The reaction was carried out at 65°C and 800 rpm for 24 h. The carrier was washed by centrifugation and resuspended in DMF to obtain 10% PEG. 2000 -Mal modified ratio of S1000 solution.

[0320] S3: DCA-modified mesoporous silica. 100 mg of thiolized DCA was dissolved in 10 mL of LDM under a dry nitrogen atmosphere. 10% PEG from S2 was then added. 2000 The modified mesoporous silica was stirred and reacted for 12-24 hours. The mixture was centrifuged and washed to obtain 10% DCA-modified S1000 mesoporous silica.

[0321] Example 43: 10% PEG with triazole linkage 2000 DCA-modified S1000 mesoporous silicon

[0322] S1: Mesoporous silica dispersion. 500 mg of the non-absorbent mesoporous silica carrier S1000 prepared in Example 2 was dispersed in a mixture of 340 mL water and 200 mL ethanol, and the pH was adjusted to 4.0 with dilute hydrochloric acid.

[0323] S2: 10% PEG 2000 -N3 modification. 10% silane-PEG was added to S1. 2000 -N3 ethanol solution was added, and then ethanol solution was added to make the ethanol:water volume ratio of the reaction system 1:2. The reaction was carried out at 65℃ and 800rpm for 24h. The carrier was washed by centrifugation and resuspended in DMF to obtain 10% PEG. 2000-N3 modified S1000 solution.

[0324] S3: DCA-modified mesoporous silica. Under a dry nitrogen atmosphere, 100 mg of alkynylated DCA, 5 mg of CuSO4·5H2O, and 10 mg of sodium ascorbate were dissolved in 5 mL of DMF. 10% PEG from S2 was then added. 2000 The modified mesoporous silica was reacted with stirring at room temperature for 12 h. The mixture was centrifuged and washed to obtain 10% DCA-modified S1000 mesoporous silica.

[0325] Example 44: 10% PEG with carbamate linkages 2000 DCA-modified S1000 mesoporous silicon

[0326] S1: Mesoporous silica dispersion. 500 mg of the non-absorbent mesoporous silica carrier S1000 prepared in Example 2 was dispersed in a mixture of 340 mL water and 200 mL ethanol, and the pH was adjusted to 4.0 with dilute hydrochloric acid.

[0327] S2: 10% PEG 2000 -NCO modification. Add 10% silane-PEG to S1. 2000 -NCO ethanol solution was added, and then ethanol solution was added to make the ethanol:water volume ratio of the reaction system 1:2. The reaction was carried out at 65℃ and 800rpm for 24h. The carrier was washed by centrifugation and resuspended in DMF to obtain 10% PEG. 2000 -NCO modified S1000 solution.

[0328] S3: DCA-modified mesoporous silica. 100 mg of isocyanated DCA was dissolved in 5 mL of LDM under a dry nitrogen atmosphere. 10 mg of DMAP was added as a catalyst, along with 10% PEG from S2. 2000 The modified mesoporous silica was reacted with stirring at room temperature for 6-12 hours. The mixture was centrifuged and washed to obtain 10% DCA-modified S1000 mesoporous silica.

[0329] Example 45: 20% PEG with amide bonds 1000 -DCA-modified S1000 mesoporous silica (S1000-20% PEG) 1000 -DCA)

[0330] S1: Mesoporous silica dispersion. 500 mg of the non-absorbent mesoporous silica carrier S1000 prepared in Example 2 was dispersed in a mixture of 340 mL water and 200 mL ethanol, and the pH was adjusted to 4.0 with dilute hydrochloric acid.

[0331] S2: 20% PEG 1000-NH2 modification. 20% silane-PEG was added to S1. 1000 The NH2 ethanol solution was added, and then ethanol solution was added to make the ethanol:water volume ratio of the reaction system 1:2. The reaction was carried out at 65℃ and 800rpm for 24h. The carrier was washed by centrifugation and resuspended in anhydrous ethanol to obtain 20% PEG. 1000- NH2-modified S1000 solution.

[0332] S3: DCA carboxyl activation. Dissolve 109.8 mg DCA in 11.0 mL DMF. Then add 21 mg NHS and 56 mg EDC sequentially, activate at room temperature for 15 min, and adjust pH to 5.8.

[0333] S4: DCA-modified mesoporous silica. The activation solution of S3 was added to 50 mL of ethanol solution (10 mg / mL). The reaction was carried out at room temperature and 500 rpm in the dark for 24 h. The mixture was centrifuged and washed to obtain 20% DCA-modified S1000 mesoporous silica. S1000-20% PEG 1000 and S1000-20% PEG 1000 - The particle size and potential of DCA are as follows Figure 2 As shown, the surface potential gradually approaches electroneutrality from electronegativity, and then becomes electronegative again after DCA grafting. This may be because the carboxyl groups of DCA bind to the amino groups on the PEG surface, weakening the charge shielding effect of PEG itself. Characterization of the potential demonstrates the successful modification with DCA.

[0334] Example 46: 20% PEG with ester bonds 1000 DCA-modified S1000 mesoporous silicon

[0335] S1: Mesoporous silica dispersion. 500 mg of the non-absorbent mesoporous silica carrier S1000 prepared in Example 2 was dispersed in a mixture of 340 mL water and 200 mL ethanol, and the pH was adjusted to 4.0 with dilute hydrochloric acid.

[0336] S2: 20% PEG 1000 -OH modification. Add 20% silane-PEG to S1. 1000 -OH ethanol solution was added, and then ethanol solution was added to make the ethanol:water volume ratio of the reaction system 1:2. The reaction was carried out at 65℃ and 800rpm for 24h. The carrier was washed by centrifugation and resuspended in anhydrous ethanol to obtain 20% PEG. 1000- OH-modified S1000 solution.

[0337] S3: DCA carboxyl activation. Under a dry nitrogen atmosphere, 100 mg DCA was dissolved in 5 mL DMF. 125 mg DCC was added, and the mixture was stirred at room temperature for 1 h to form the activated intermediate O-acylurea.

[0338] S4: DCA-modified mesoporous silica. The intermediate from S3 was added to 5 mg of DPTS, followed by the addition of 20% PEG from S2. 1000 The modified mesoporous silica was stirred and reacted for 12-24 hours. The mixture was centrifuged and washed to obtain 20% DCA-modified S1000 mesoporous silica.

[0339] Example 47: 20% PEG1000-DCA modified S1000 mesoporous silica with thioether bonds

[0340] S1: Mesoporous silica dispersion. 500 mg of the non-absorbent mesoporous silica carrier S1000 prepared in Example 2 was dispersed in a mixture of 340 mL water and 200 mL ethanol, and the pH was adjusted to 4.0 with dilute hydrochloric acid.

[0341] S2: 20% PEG 1000 -Mal modification. Add 20% silane-PEG to S1. 1000 The ethanol solution was first prepared, and then ethanol was added to make the ethanol:water volume ratio of the reaction system 1:2. The reaction was carried out at 65°C and 800 rpm for 24 h. The carrier was washed by centrifugation and resuspended in DMF to obtain 20% PEG. 1000 -Mal-modified S1000 solution.

[0342] S3: DCA-modified mesoporous silica. 100 mg of thiolized DCA was dissolved in 10 mL of LDM under a dry nitrogen atmosphere. 20% PEG from S2 was then added. 1000 The modified mesoporous silica was stirred and reacted for 12-24 hours. The mixture was centrifuged and washed to obtain 20% DCA-modified S1000 mesoporous silica.

[0343] Example 48: 20% PEG with triazole linkage 1000 DCA-modified S1000 mesoporous silicon

[0344] S1: Mesoporous silica dispersion. 500 mg of the non-absorbent mesoporous silica carrier S1000 prepared in Example 2 was dispersed in a mixture of 340 mL water and 200 mL ethanol, and the pH was adjusted to 4.0 with dilute hydrochloric acid.

[0345] S2: 20% PEG 1000 -N3 modification. 20% silane-PEG was added to S1. 1000-N3 ethanol solution was added, and then ethanol solution was added to make the ethanol:water volume ratio of the reaction system 1:2. The reaction was carried out at 65℃ and 800rpm for 24h. The carrier was washed by centrifugation and resuspended in DMF to obtain 20% PEG. 1000 -N3 modified ratio of S1000 solution.

[0346] S3: DCA-modified mesoporous silica. Under a dry nitrogen atmosphere, 100 mg of alkynylated DCA, 5 mg of CuSO4·5H2O, and 10 mg of sodium ascorbate were dissolved in 5 mL of LDMF. 20% of the PEG from S2 was then added. 1000 The modified mesoporous silica was reacted with stirring at room temperature for 12 h. The mixture was centrifuged and washed to obtain 20% DCA-modified S1000 mesoporous silica.

[0347] Example 49: 20% PEG with carbamate bonds 1000 DCA-modified S1000 mesoporous silicon

[0348] S1: Mesoporous silica dispersion. 500 mg of the non-absorbent mesoporous silica carrier S1000 prepared in Example 2 was dispersed in a mixture of 340 mL water and 200 mL ethanol, and the pH was adjusted to 4.0 with dilute hydrochloric acid.

[0349] S2: 20% PEG 1000 -NCO modification. Add 20% silane-PEG to S1. 1000 -NCO ethanol solution was added, and then ethanol solution was added to make the ethanol:water volume ratio of the reaction system 1:2. The reaction was carried out at 65℃ and 800rpm for 24h. The carrier was washed by centrifugation and resuspended in DMF to obtain 20% PEG. 1000 -NCO modified S1000 solution.

[0350] S3: DCA-modified mesoporous silica. 100 mg of isocyanated DCA was dissolved in 5 mL of LDM under a dry nitrogen atmosphere. 10 mg of DMAP was added as a catalyst, along with 20% PEG from S2. 1000 The modified mesoporous silica was reacted with stirring at room temperature for 6-12 h. The mixture was centrifuged and washed to obtain 20% DCA-modified S1000 mesoporous silica.

[0351] Example 50: 20% PEG with amide bonds 2000 -DCA-modified S1000 mesoporous silica (S1000-20% PEG) 2000 -DCA)

[0352] S1: Mesoporous silica dispersion. 500 mg of the non-absorbent mesoporous silica carrier S1000 prepared in Example 2 was dispersed in a mixture of 340 mL water and 200 mL ethanol, and the pH was adjusted to 4.0 with dilute hydrochloric acid.

[0353] S2: 20% PEG 2000 -NH2 modification. 20% silane-PEG was added to S1. 2000 The NH2 ethanol solution was added, and then ethanol solution was added to make the ethanol:water volume ratio of the reaction system 1:2. The reaction was carried out at 65℃ and 800rpm for 24h. The carrier was washed by centrifugation and resuspended in anhydrous ethanol to obtain 20% PEG. 2000 S1000 solution with -NH2 modification ratio.

[0354] S3: DCA carboxyl activation. Dissolve 109.8 mg DCA in 11.0 mL DMF. Then add 21 mg NHS and 56 mg EDC sequentially, activate at room temperature for 15 min, and adjust pH to 5.8.

[0355] S4: DCA-modified mesoporous silica. The activation solution of S3 was added to 50 mL of ethanol solution (10 mg / mL). The reaction was carried out at room temperature and 500 rpm in the dark for 24 h. The mixture was centrifuged and washed to obtain 20% DCA-modified S1000 mesoporous silica. S1000-20% PEG 2000 and S1000-20% PEG 2000 - The particle size and potential of DCA are as follows Figure 2 As shown, the surface potential gradually approaches electroneutrality from electronegativity, and then becomes electronegative again after DCA grafting. This may be because the carboxyl groups of DCA bind to the amino groups on the PEG surface, weakening the charge shielding effect of PEG itself. Characterization of the potential demonstrates the successful modification with DCA.

[0356] Example 51: 20% PEG with ester bonds 2000 DCA-modified S1000 mesoporous silicon

[0357] S1: Mesoporous silica dispersion. 500 mg of the non-absorbent mesoporous silica carrier S1000 prepared in Example 2 was dispersed in a mixture of 340 mL water and 200 mL ethanol, and the pH was adjusted to 4.0 with dilute hydrochloric acid.

[0358] S2: 20% PEG 2000 -OH modification. Add 20% silane-PEG to S1. 2000-OH ethanol solution was added, and then ethanol solution was added to make the ethanol:water volume ratio of the reaction system 1:2. The reaction was carried out at 65℃ and 800rpm for 24h. The carrier was washed by centrifugation and resuspended in anhydrous ethanol to obtain 20% PEG. 2000 S1000 solution with -OH modification ratio.

[0359] S3: DCA carboxyl activation. Under a dry nitrogen atmosphere, 100 mg DCA was dissolved in 5 mL DMF. 125 mg DCC was added, and the mixture was stirred at room temperature for 1 h to form the activated intermediate O-acylurea.

[0360] S4: DCA-modified mesoporous silica. 5 mg of DPTS was added to the intermediate in S3, followed by the addition of 20% PEG from S2. 2000 The modified mesoporous silica was stirred and reacted for 12-24 hours. The mixture was centrifuged and washed to obtain 20% DCA-modified S1000 mesoporous silica.

[0361] Example 52: 20% PEG with thioether linkages 2000 DCA-modified S1000 mesoporous silicon

[0362] S1: Mesoporous silica dispersion. 500 mg of the non-absorbent mesoporous silica carrier S1000 prepared in Example 2 was dispersed in a mixture of 340 mL water and 200 mL ethanol, and the pH was adjusted to 4.0 with dilute hydrochloric acid.

[0363] S2: 20% PEG 2000 -Mal modification. Add 20% silane-PEG to S1. 2000 The ethanol solution was first prepared, and then ethanol was added to make the ethanol:water volume ratio of the reaction system 1:2. The reaction was carried out at 65°C and 800 rpm for 24 h. The carrier was washed by centrifugation and resuspended in DMF to obtain 20% PEG. 2000 -Mal modified ratio of S1000 solution.

[0364] S3: DCA-modified mesoporous silica. 100 mg of thiolized DCA was dissolved in 10 mL of LDM under a dry nitrogen atmosphere. 20% PEG from S2 was then added. 2000 The modified mesoporous silica was stirred and reacted for 12-24 hours. The mixture was centrifuged and washed to obtain 20% DCA-modified S1000 mesoporous silica.

[0365] Example 53: 20% PEG with triazole linkage 2000 DCA-modified S1000 mesoporous silicon

[0366] S1: Mesoporous silica dispersion. 500 mg of the non-absorbent mesoporous silica carrier S1000 prepared in Example 2 was dispersed in a mixture of 340 mL water and 200 mL ethanol, and the pH was adjusted to 4.0 with dilute hydrochloric acid.

[0367] S2: 20% PEG 2000 -N3 modification. 20% silane-PEG was added to S1. 2000 -N3 ethanol solution was added, and then ethanol solution was added to make the ethanol:water volume ratio of the reaction system 1:2. The reaction was carried out at 65℃ and 800rpm for 24h. The carrier was washed by centrifugation and resuspended in DMF to obtain 20% PEG. 2000 -N3 modified S1000 solution.

[0368] S3: DCA-modified mesoporous silica. Under a dry nitrogen atmosphere, 100 mg of alkynylated DCA, 5 mg of CuSO4·5H2O, and 10 mg of sodium ascorbate were dissolved in 5 mL of DMF. 20% of the PEG from S2 was then added. 2000 The modified mesoporous silica was reacted with stirring at room temperature for 12 h. The mixture was centrifuged and washed to obtain 20% DCA-modified S1000 mesoporous silica.

[0369] Example 54: 20% PEG with carbamate bonds 2000 DCA-modified S1000 mesoporous silicon

[0370] S1: Mesoporous silica dispersion. 500 mg of the non-absorbent mesoporous silica carrier S1000 prepared in Example 2 was dispersed in a mixture of 340 mL water and 200 mL ethanol, and the pH was adjusted to 4.0 with dilute hydrochloric acid.

[0371] S2: 20% PEG 2000 -NCO modification. Add 20% silane-PEG to S1. 2000 -NCO ethanol solution was added, and then ethanol solution was added to make the ethanol:water volume ratio of the reaction system 1:2. The reaction was carried out at 65℃ and 800rpm for 24h. The carrier was washed by centrifugation and resuspended in DMF to obtain 20% PEG. 2000 -NCO modified S1000 solution.

[0372] S3: DCA-modified mesoporous silica. 100 mg of isocyanated DCA was dissolved in 5 mL of LDM under a dry nitrogen atmosphere. 10 mg of DMAP was added as a catalyst, along with 20% PEG from S2. 2000 The modified mesoporous silica was reacted with stirring at room temperature for 6-12 h. The mixture was centrifuged and washed to obtain 20% DCA-modified S1000 mesoporous silica.

[0373] Example 55: 30% PEG with amide bonds 1000 -DCA-modified S1000 mesoporous silica (S1000-30% PEG) 1000 -DCA)

[0374] S1: Mesoporous silica dispersion. 500 mg of the non-absorbent mesoporous silica carrier S1000 prepared in Example 2 was dispersed in a mixture of 340 mL water and 200 mL ethanol, and the pH was adjusted to 4.0 with dilute hydrochloric acid.

[0375] S2: 30% PEG 1000 -NH2 modification. 30% silane-PEG was added to S1. 1000 -NH2 ethanol solution was added, and then ethanol solution was added to make the ethanol:water volume ratio of the reaction system 1:2. The reaction was carried out at 65℃ and 800rpm for 24h. The carrier was washed by centrifugation and resuspended in anhydrous ethanol to obtain 30% PEG. 1000- NH2-modified S1000 solution.

[0376] S3: DCA carboxyl activation. Dissolve 109.8 mg DCA in 11.0 mL DMF. Then add 21 mg NHS and 56 mg EDC sequentially, activate at room temperature for 15 min, and adjust pH to 5.8.

[0377] S4: DCA-modified mesoporous silica. The activation solution of S3 was added to 50 mL of an ethanol solution of S2 (10 mg / mL). The reaction was carried out at room temperature and 500 rpm in the dark for 24 h. The mixture was centrifuged and washed to obtain 30% DCA-modified S1000 mesoporous silica. S1000-30% PEG 1000 and S1000-30% PEG 1000 - The particle size and potential of DCA are as follows Figure 2 As shown, the surface potential gradually approaches electroneutrality from electronegativity. After DCA grafting, S1000-30% PEG 1000 -DCA becomes electronegative again, possibly because the carboxyl group of DCA binds to the amino group on the PEG surface, weakening the charge shielding effect of PEG itself. Characterization of the potential preliminarily confirms the successful modification of DCA.

[0378] Example 56: 30% PEG with ester bonds 1000 DCA-modified S1000 mesoporous silicon

[0379] S1: Mesoporous silica dispersion. 500 mg of the non-absorbent mesoporous silica carrier S1000 prepared in Example 2 was dispersed in a mixture of 340 mL water and 200 mL ethanol, and the pH was adjusted to 4.0 with dilute hydrochloric acid.

[0380] S2: 30% PEG 1000 -OH modification. Add 30% silane-PEG to S1. 1000 -OH ethanol solution was added, and then ethanol solution was added to make the ethanol:water volume ratio of the reaction system 1:2. The reaction was carried out at 65℃ and 800rpm for 24h. The carrier was washed by centrifugation and resuspended in anhydrous ethanol to obtain 30% PEG. 1000- OH-modified S1000 solution.

[0381] S3: DCA carboxyl activation. Under a dry nitrogen atmosphere, 100 mg DCA was dissolved in 5 mL DMF. 125 mg DCC was added, and the mixture was stirred at room temperature for 1 h to form the activated intermediate O-acylurea.

[0382] S4: DCA-modified mesoporous silica. The intermediate from S3 was added to 5 mg of DPTS, followed by the addition of 30% PEG from S2. 1000 The modified mesoporous silica was stirred and reacted for 12-24 hours. The mixture was centrifuged and washed to obtain 30% DCA-modified S1000 mesoporous silica.

[0383] Example 57: 30% PEG1000-DCA modified S1000 mesoporous silica with thioether bonds

[0384] S1: Mesoporous silica dispersion. 500 mg of the non-absorbent mesoporous silica carrier S1000 prepared in Example 2 was dispersed in a mixture of 340 mL water and 200 mL ethanol, and the pH was adjusted to 4.0 with dilute hydrochloric acid.

[0385] S2: 30% PEG 1000 -Mal modification. Add 30% silane-PEG to S1. 1000 The ethanol solution was first prepared, and then ethanol was added to make the ethanol:water volume ratio of the reaction system 1:2. The reaction was carried out at 65°C and 800 rpm for 24 h. The carrier was washed by centrifugation and resuspended in DMF to obtain 30% PEG. 1000 -Mal-modified S1000 solution.

[0386] S3: DCA-modified mesoporous silica. 100 mg of thiolized DCA was dissolved in 10 mL of LDM under a dry nitrogen atmosphere. 30% of the PEG from S2 was then added. 1000 The modified mesoporous silica was stirred and reacted for 12-24 hours. The mixture was centrifuged and washed to obtain 30% DCA-modified S1000 mesoporous silica.

[0387] Example 58: 30% PEG with triazole linkage 1000DCA-modified S1000 mesoporous silicon

[0388] S1: Mesoporous silica dispersion. 500 mg of the non-absorbent mesoporous silica carrier S1000 prepared in Example 2 was dispersed in a mixture of 340 mL water and 200 mL ethanol, and the pH was adjusted to 4.0 with dilute hydrochloric acid.

[0389] S2: 30% PEG 1000 -N3 modification. 30% silane-PEG was added to S1. 1000 -N3 ethanol solution was added, and then ethanol solution was added to make the ethanol:water volume ratio of the reaction system 1:2. The reaction was carried out at 65℃ and 800rpm for 24h. The carrier was washed by centrifugation and resuspended in DMF to obtain 30% PEG. 1000 -N3 modified ratio of S1000 solution.

[0390] S3: DCA-modified mesoporous silica. Under a dry nitrogen atmosphere, 100 mg of alkynylated DCA, 5 mg of CuSO4·5H2O, and 10 mg of sodium ascorbate were dissolved in 5 mL of DMF. 30% of the PEG from S2 was then added. 1000 The modified mesoporous silica was reacted with stirring at room temperature for 12 h. The mixture was centrifuged and washed to obtain 30% DCA-modified S1000 mesoporous silica.

[0391] Example 59: 30% PEG with carbamate bonds 1000 DCA-modified S1000 mesoporous silicon

[0392] S1: Mesoporous silica dispersion. 500 mg of the non-absorbent mesoporous silica carrier S1000 prepared in Example 2 was dispersed in a mixture of 340 mL water and 200 mL ethanol, and the pH was adjusted to 4.0 with dilute hydrochloric acid.

[0393] S2: 30% PEG 1000 -NCO modification. Add 30% silane-PEG to S1. 1000 -NCO ethanol solution was added, and then ethanol solution was added to make the ethanol:water volume ratio of the reaction system 1:2. The reaction was carried out at 65℃ and 800rpm for 24h. The carrier was washed by centrifugation and resuspended in DMF to obtain 30% PEG. 1000 -NCO modified S1000 solution.

[0394] S3: DCA-modified mesoporous silica. 100 mg of isocyanated DCA was dissolved in 5 mL of LDM under a dry nitrogen atmosphere. 10 mg of DMAP was added as a catalyst, along with 30% PEG from S2. 1000The modified mesoporous silica was reacted with stirring at room temperature for 6-12 h. The mixture was centrifuged and washed to obtain 30% DCA-modified S1000 mesoporous silica.

[0395] Example 60: 30% PEG with amide bonds 2000 -DCA-modified S1000 mesoporous silica (S1000-30% PEG) 2000 -DCA)

[0396] S1: Mesoporous silica dispersion. 500 mg of the non-absorbent mesoporous silica carrier S1000 prepared in Example 2 was dispersed in a mixture of 340 mL water and 200 mL ethanol, and the pH was adjusted to 4.0 with dilute hydrochloric acid.

[0397] S2: 30% PEG 2000 -NH2 modification. 30% silane-PEG was added to S1. 2000 -NH2 ethanol solution was added, and then ethanol solution was added to make the ethanol:water volume ratio of the reaction system 1:2. The reaction was carried out at 65℃ and 800rpm for 24h. The carrier was washed by centrifugation and resuspended in anhydrous ethanol to obtain 30% PEG. 2000 S1000 solution with -NH2 modification ratio.

[0398] S3: DCA carboxyl activation. Dissolve 109.8 mg DCA in 11.0 mL DMF. Then add 21 mg NHS and 56 mg EDC sequentially, activate at room temperature for 15 min, and adjust pH to 5.8.

[0399] S4: DCA-modified mesoporous silica. The activation solution of S3 was added to 50 mL of ethanol solution (10 mg / mL). The reaction was carried out at room temperature and 500 rpm in the dark for 24 h. The mixture was centrifuged and washed to obtain 30% DCA-modified S1000 mesoporous silica. S1000-30% PEG 2000 and S1000-30% PEG 2000 - The particle size and potential of DCA are as follows Figure 2 As shown, the surface potential gradually approaches electroneutrality from electronegativity, and then becomes electronegative again after DCA grafting. This may be because the carboxyl groups of DCA bind to the amino groups on the PEG surface, weakening the charge shielding effect of PEG itself. Characterization of the potential demonstrates the successful modification with DCA.

[0400] Example 61: 30% PEG with ester bonds 2000 DCA-modified S1000 mesoporous silicon

[0401] S1: Mesoporous silica dispersion. 500 mg of the non-absorbent mesoporous silica carrier S1000 prepared in Example 2 was dispersed in a mixture of 340 mL water and 200 mL ethanol, and the pH was adjusted to 4.0 with dilute hydrochloric acid.

[0402] S2: 30% PEG 2000 -OH modification. Add 30% silane-PEG to S1. 2000 -OH ethanol solution was added, and then ethanol solution was added to make the ethanol:water volume ratio of the reaction system 1:2. The reaction was carried out at 65℃ and 800rpm for 24h. The carrier was washed by centrifugation and resuspended in anhydrous ethanol to obtain 30% PEG. 2000 S1000 solution with -OH modification ratio.

[0403] S3: DCA carboxyl activation. Under a dry nitrogen atmosphere, 100 mg DCA was dissolved in 5 mL DMF. 125 mg DCC was added, and the mixture was stirred at room temperature for 1 h to form the activated intermediate O-acylurea.

[0404] S4: DCA-modified mesoporous silica. 5 mg of DPTS was added to the intermediate in S3, followed by the addition of 30% PEG from S2. 2000 The modified mesoporous silica was stirred and reacted for 12-24 hours. The mixture was centrifuged and washed to obtain 30% DCA-modified S1000 mesoporous silica.

[0405] Example 62: 30% PEG with thioether linkages 2000 DCA-modified S1000 mesoporous silicon

[0406] S1: Mesoporous silica dispersion. 500 mg of the non-absorbent mesoporous silica carrier S1000 prepared in Example 2 was dispersed in a mixture of 340 mL water and 200 mL ethanol, and the pH was adjusted to 4.0 with dilute hydrochloric acid.

[0407] S2: 30% PEG 2000 -Mal modification. Add 30% silane-PEG to S1. 2000 The ethanol solution was first prepared, and then ethanol was added to make the ethanol:water volume ratio of the reaction system 1:2. The reaction was carried out at 65°C and 800 rpm for 24 h. The carrier was washed by centrifugation and resuspended in DMF to obtain 30% PEG. 2000 -Mal modified ratio of S1000 solution.

[0408] S3: DCA-modified mesoporous silica. 100 mg of thiolized DCA was dissolved in 10 mL of LDM under a dry nitrogen atmosphere. 30% of the PEG from S2 was then added. 2000The modified mesoporous silica was stirred and reacted for 12-24 hours. The mixture was centrifuged and washed to obtain 30% DCA-modified S1000 mesoporous silica.

[0409] Example 63: 30% PEG with triazole linkage 2000 DCA-modified S1000 mesoporous silicon

[0410] S1: Mesoporous silica dispersion. 500 mg of the non-absorbent mesoporous silica carrier S1000 prepared in Example 2 was dispersed in a mixture of 340 mL water and 200 mL ethanol, and the pH was adjusted to 4.0 with dilute hydrochloric acid.

[0411] S2: 30% PEG 2000 -N3 modification. 30% silane-PEG was added to S1. 2000 -N3 ethanol solution was added, and then ethanol solution was added to make the ethanol:water volume ratio of the reaction system 1:2. The reaction was carried out at 65℃ and 800rpm for 24h. The carrier was washed by centrifugation and resuspended in DMF to obtain 30% PEG. 2000 -N3 modified S1000 solution.

[0412] S3: DCA-modified mesoporous silica. Under a dry nitrogen atmosphere, 100 mg of alkynylated DCA, 5 mg of CuSO4·5H2O, and 10 mg of sodium ascorbate were dissolved in 5 mL of DMF. 30% of the PEG from S2 was then added. 2000 The modified mesoporous silica was reacted with stirring at room temperature for 12 h. The mixture was centrifuged and washed to obtain 30% DCA-modified S1000 mesoporous silica.

[0413] Example 64: 30% PEG with carbamate bonds 2000 DCA-modified S1000 mesoporous silicon

[0414] S1: Mesoporous silica dispersion. 500 mg of the non-absorbent mesoporous silica carrier S1000 prepared in Example 2 was dispersed in a mixture of 340 mL water and 200 mL ethanol, and the pH was adjusted to 4.0 with dilute hydrochloric acid.

[0415] S2: 30% PEG 2000 -NCO modification. Add 30% silane-PEG to S1. 2000 -NCO ethanol solution was added, and then ethanol solution was added to make the ethanol:water volume ratio of the reaction system 1:2. The reaction was carried out at 65℃ and 800rpm for 24h. The carrier was washed by centrifugation and resuspended in DMF to obtain 30% PEG. 2000 -NCO modified S1000 solution.

[0416] S3: DCA-modified mesoporous silica. 100 mg of isocyanated DCA was dissolved in 5 mL of LDM under a dry nitrogen atmosphere. 10 mg of DMAP was added as a catalyst, along with 30% PEG from S2. 2000 The modified mesoporous silica was reacted with stirring at room temperature for 6-12 h. The mixture was centrifuged and washed to obtain 30% DCA-modified S1000 mesoporous silica.

[0417] Example 65: 10% PEG with amide bonds 1000 -DCA-modified R500 mesoporous silicon (R500-10% PEG) 1000 -DCA)

[0418] S1: Mesoporous silica dispersion. 500 mg of the non-absorbent mesoporous silica support R500 prepared in Example 3 was dispersed in a mixture of 340 mL water and 200 mL ethanol, and the pH was adjusted to 4.0 with dilute hydrochloric acid.

[0419] S2: 10% PEG 1000 -NH2 modification. 10% silane-PEG was added to S1. 1000 The NH2 ethanol solution was added, and then ethanol solution was added to make the ethanol:water volume ratio of the reaction system 1:2. The reaction was carried out at 65℃ and 800rpm for 24h. The carrier was washed by centrifugation and resuspended in anhydrous ethanol to obtain 10% PEG. 1000 -NH2 modified R500 solution.

[0420] S3: DCA carboxyl activation. Dissolve 109.8 mg DCA in 11.0 mL DMF. Then add 21 mg NHS and 56 mg EDC sequentially, activate at room temperature for 15 min, and adjust pH to 5.8.

[0421] S4: DCA-modified mesoporous silica. The activation solution of S3 was added to 50 mL of ethanol solution (10 mg / mL). The reaction was carried out at room temperature and 500 rpm in the dark for 24 h. The mixture was centrifuged and washed to obtain 10% DCA-modified R500 mesoporous silica. R500-10% PEG 1000 and R500-10% PEG 1000 - The particle size and potential of DCA are as follows Figure 2 As shown, the surface potential gradually approaches electroneutrality from electronegativity, and then becomes electronegative again after DCA grafting. This may be because the carboxyl groups of DCA bind to the amino groups on the PEG surface, weakening the charge shielding effect of PEG itself. Characterization of the potential demonstrates the successful modification with DCA.

[0422] Example 66: 10% PEG with ester bonds 1000-DCA-modified R500 mesoporous silica (R500-10% PEG) 1000 -DCA)

[0423] S1: Mesoporous silica dispersion. 500 mg of the non-absorbent mesoporous silica support R500 prepared in Example 3 was dispersed in a mixture of 340 mL water and 200 mL ethanol, and the pH was adjusted to 4.0 with dilute hydrochloric acid.

[0424] S2: 10% PEG 1000 -OH modification. Add 10% silane-PEG to S1. 1000 -OH ethanol solution was added, and then ethanol solution was added to make the ethanol:water volume ratio of the reaction system 1:2. The reaction was carried out at 65℃ and 800rpm for 24h. The carrier was washed by centrifugation and resuspended in anhydrous ethanol to obtain 10% PEG. 1000 R500 solution modified with -OH.

[0425] S3: DCA carboxyl activation. Under a dry nitrogen atmosphere, 100 mg DCA was dissolved in 5 mL DMF. 125 mg DCC was added, and the mixture was stirred at room temperature for 1 h to form the activated intermediate O-acylurea.

[0426] S4: DCA-modified mesoporous silica. 5 mg of DPTS was added to the intermediate from S3, followed by the addition of 10% PEG from S2. 1000 Modified mesoporous silica was reacted with stirring for 12-24 h. The mixture was centrifuged and washed to obtain 10% DCA-modified R500 mesoporous silica. Results are as follows: Figure 4 As shown, R500-10% PEG 1000 -The DCA appears as a long rod under an electron microscope, with a length and width of approximately 850 nm and 40 nm, respectively.

[0427] Example 67: 10% PEG with thioether linkages 1000 DCA-modified R500 mesoporous silicon

[0428] S1: Mesoporous silica dispersion. 500 mg of the non-absorbent mesoporous silica support R500 prepared in Example 3 was dispersed in a mixture of 340 mL water and 200 mL ethanol, and the pH was adjusted to 4.0 with dilute hydrochloric acid.

[0429] S2: 10% PEG 1000 -Mal modification. Add 10% silane-PEG to S1. 1000 The ethanol solution was first prepared, and then ethanol was added to make the ethanol:water volume ratio of the reaction system 1:2. The reaction was carried out at 65°C and 800 rpm for 24 h. The carrier was washed by centrifugation and resuspended in DMF to obtain 10% PEG. 1000-Mal-modified R500 solution.

[0430] S3: DCA-modified mesoporous silica. 100 mg of thiolized DCA was dissolved in 10 mL of LDM under a dry nitrogen atmosphere. 10% PEG from S2 was then added. 1000 The modified mesoporous silica was stirred and reacted for 12-24 hours. The mixture was centrifuged and washed to obtain 10% DCA-modified R500 mesoporous silica.

[0431] Example 68: 10% PEG with triazole linkage 1000 DCA-modified R500 mesoporous silicon

[0432] S1: Mesoporous silica dispersion. 500 mg of the non-absorbent mesoporous silica support R500 prepared in Example 3 was dispersed in a mixture of 340 mL water and 200 mL ethanol, and the pH was adjusted to 4.0 with dilute hydrochloric acid.

[0433] S2: 10% PEG 1000 -N3 modification. 10% silane-PEG was added to S1. 1000 -N3 ethanol solution was added, and then ethanol solution was added to make the ethanol:water volume ratio of the reaction system 1:2. The reaction was carried out at 65℃ and 800rpm for 24h. The carrier was washed by centrifugation and resuspended in DMF to obtain 10% PEG. 1000 -N3 modified R500 solution.

[0434] S3: DCA-modified mesoporous silica. Under a dry nitrogen atmosphere, 100 mg of alkynylated DCA, 5 mg of CuSO4·5H2O, and 10 mg of sodium ascorbate were dissolved in 5 mL of DMF. 10% PEG from S2 was then added. 1000 The modified mesoporous silica was reacted with stirring at room temperature for 12 h. The mixture was centrifuged and washed to obtain 10% DCA-modified R500 mesoporous silica.

[0435] Example 69: 10% PEG with carbamate bonds 1000 DCA-modified R500 mesoporous silicon

[0436] S1: Mesoporous silica dispersion. 500 mg of the non-absorbent mesoporous silica support R500 prepared in Example 3 was dispersed in a mixture of 340 mL water and 200 mL ethanol, and the pH was adjusted to 4.0 with dilute hydrochloric acid.

[0437] S2: 10% PEG 1000 -NCO modification. Add 10% silane-PEG to S1. 1000-NCO ethanol solution was added, and then ethanol solution was added to make the ethanol:water volume ratio of the reaction system 1:2. The reaction was carried out at 65℃ and 800rpm for 24h. The carrier was washed by centrifugation and resuspended in DMF to obtain 10% PEG. 1000 -NCO modified R500 solution.

[0438] S3: DCA-modified mesoporous silica. 100 mg of isocyanated DCA was dissolved in 5 mL of LDM under a dry nitrogen atmosphere. 10 mg of DMAP was added as a catalyst, along with 10% PEG from S2. 1000 The modified mesoporous silica was reacted with stirring at room temperature for 6-12 h. The mixture was centrifuged and washed to obtain 10% DCA-modified R500 mesoporous silica.

[0439] Example 70: 20% PEG with amide bonds 1000 -DCA-modified R500 mesoporous silica (R500-20% PEG) 1000 -DCA)

[0440] S1: Mesoporous silica dispersion. 500 mg of the non-absorbent mesoporous silica support R500 prepared in Example 3 was dispersed in a mixture of 340 mL water and 200 mL ethanol, and the pH was adjusted to 4.0 with dilute hydrochloric acid.

[0441] S2: 20% PEG 1000 -NH2 modification. 20% silane-PEG was added to S1. 1000 The NH2 ethanol solution was added, and then ethanol solution was added to make the ethanol:water volume ratio of the reaction system 1:2. The reaction was carried out at 65℃ and 800rpm for 24h. The carrier was washed by centrifugation and resuspended in anhydrous ethanol to obtain 20% PEG. 1000 -NH2 modified R500 solution.

[0442] S3: DCA carboxyl activation. Dissolve 109.8 mg DCA in 11.0 mL DMF. Then add 21 mg NHS and 56 mg EDC sequentially, activate at room temperature for 15 min, and adjust pH to 5.8.

[0443] S4: DCA-modified mesoporous silica. The activation solution of S3 was added to 50 mL of ethanol solution (10 mg / mL). The reaction was carried out at room temperature and 500 rpm in the dark for 24 h. The mixture was centrifuged and washed to obtain 20% DCA-modified R500 mesoporous silica. R500-20% PEG 1000 and R500-20% PEG 1000 - The particle size and potential of DCA are as follows Figure 2As shown, the surface potential gradually approaches electroneutrality from electronegativity, and then becomes electronegative again after DCA grafting. This may be because the carboxyl groups of DCA bind to the amino groups on the PEG surface, weakening the charge shielding effect of PEG itself. Characterization of the potential demonstrates the successful modification with DCA.

[0444] Example 71: 20% PEG with ester bonds 1000 DCA-modified R500 mesoporous silicon

[0445] S1: Mesoporous silica dispersion. 500 mg of the non-absorbent mesoporous silica support R500 prepared in Example 3 was dispersed in a mixture of 340 mL water and 200 mL ethanol, and the pH was adjusted to 4.0 with dilute hydrochloric acid.

[0446] S2: 20% PEG 1000 -OH modification. Add 20% silane-PEG to S1. 1000 -OH ethanol solution was added, and then ethanol solution was added to make the ethanol:water volume ratio of the reaction system 1:2. The reaction was carried out at 65℃ and 800rpm for 24h. The carrier was washed by centrifugation and resuspended in anhydrous ethanol to obtain 20% PEG. 1000 R500 solution modified with -OH.

[0447] S3: DCA carboxyl activation. Under a dry nitrogen atmosphere, 100 mg DCA was dissolved in 5 mL DMF. 125 mg DCC was added, and the mixture was stirred at room temperature for 1 h to form the activated intermediate O-acylurea.

[0448] S4: DCA-modified mesoporous silica. 5 mg of DPTS was added to the intermediate of S3, followed by the addition of 20% PEG from S2. 1000 The modified mesoporous silica was stirred and reacted for 12-24 hours. The mixture was centrifuged and washed to obtain 20% DCA-modified R500 mesoporous silica.

[0449] Example 72, 20% PEG with thioether linkage 1000 DCA-modified R500 mesoporous silicon

[0450] S1: Mesoporous silica dispersion. 500 mg of the non-absorbent mesoporous silica support R500 prepared in Example 3 was dispersed in a mixture of 340 mL water and 200 mL ethanol, and the pH was adjusted to 4.0 with dilute hydrochloric acid.

[0451] S2: 20% PEG 1000 -Mal modification. Add 20% silane-PEG to S1. 1000The ethanol solution was first prepared, and then ethanol was added to make the ethanol:water volume ratio of the reaction system 1:2. The reaction was carried out at 65°C and 800 rpm for 24 h. The carrier was washed by centrifugation and resuspended in DMF to obtain 20% PEG. 1000 -Mal-modified R500 solution.

[0452] S3: DCA-modified mesoporous silica. 100 mg of thiolized DCA was dissolved in 10 mL of LDM under a dry nitrogen atmosphere. 20% PEG from S2 was then added. 1000 The modified mesoporous silica was stirred and reacted for 12-24 hours. The mixture was centrifuged and washed to obtain 20% DCA-modified R500 mesoporous silica.

[0453] Example 73: 20% PEG with triazole linkage 1000 DCA-modified R500 mesoporous silicon

[0454] S1: Mesoporous silica dispersion. 500 mg of the non-absorbent mesoporous silica support R500 prepared in Example 3 was dispersed in a mixture of 340 mL water and 200 mL ethanol, and the pH was adjusted to 4.0 with dilute hydrochloric acid.

[0455] S2: 20% PEG 1000 -N3 modification. 20% silane-PEG was added to S1. 1000 -N3 ethanol solution was added, and then ethanol solution was added to make the ethanol:water volume ratio of the reaction system 1:2. The reaction was carried out at 65℃ and 800rpm for 24h. The carrier was washed by centrifugation and resuspended in DMF to obtain 20% PEG. 1000 -N3 modified R500 solution.

[0456] S3: DCA-modified mesoporous silica. Under a dry nitrogen atmosphere, 100 mg of alkynylated DCA, 5 mg of CuSO4·5H2O, and 10 mg of sodium ascorbate were dissolved in 5 mL of DMF. 20% of the PEG from S2 was then added. 1000 The modified mesoporous silica was reacted with stirring at room temperature for 12 h. The mixture was centrifuged and washed to obtain 20% DCA-modified R500 mesoporous silica.

[0457] Example 74: 20% PEG with carbamate bonds 1000 DCA-modified R500 mesoporous silicon

[0458] S1: Mesoporous silica dispersion. 500 mg of the non-absorbent mesoporous silica support R500 prepared in Example 3 was dispersed in a mixture of 340 mL water and 200 mL ethanol, and the pH was adjusted to 4.0 with dilute hydrochloric acid.

[0459] S2: 20% PEG 1000 -NCO modification. Add 20% silane-PEG to S1. 1000 -NCO ethanol solution was added, and then ethanol solution was added to make the ethanol:water volume ratio of the reaction system 1:2. The reaction was carried out at 65℃ and 800rpm for 24h. The carrier was washed by centrifugation and resuspended in DMF to obtain 20% PEG. 1000 -NCO modified R500 solution.

[0460] S3: DCA-modified mesoporous silica. 100 mg of isocyanated DCA was dissolved in 5 mL of LDM under a dry nitrogen atmosphere. 10 mg of DMAP was added as a catalyst, along with 20% PEG from S2. 1000 The modified mesoporous silica was reacted with stirring at room temperature for 6-12 h. The mixture was centrifuged and washed to obtain 20% DCA-modified R500 mesoporous silica.

[0461] Example 75: 30% PEG with amide bonds 1000 -DCA-modified R500 mesoporous silicon (R500-30% PEG) 1000 -DCA)

[0462] S1: Mesoporous silica dispersion. 500 mg of the non-absorbent mesoporous silica support R500 prepared in Example 3 was dispersed in a mixture of 340 mL water and 200 mL ethanol, and the pH was adjusted to 4.0 with dilute hydrochloric acid.

[0463] S2: 30% PEG 1000 -NH2 modification. 30% silane-PEG was added to S1. 1000 -NH2 ethanol solution was added, and then ethanol solution was added to make the ethanol:water volume ratio of the reaction system 1:2. The reaction was carried out at 65℃ and 800rpm for 24h. The carrier was washed by centrifugation and resuspended in anhydrous ethanol to obtain 30% PEG. 1000 -NH2 modified R500 solution.

[0464] S3: DCA carboxyl activation. Dissolve 109.8 mg DCA in 11.0 mL DMF. Then add 21 mg NHS and 56 mg EDC sequentially, activate at room temperature for 15 min, and adjust pH to 5.8.

[0465] S4: DCA-modified mesoporous silica. The activation solution of S3 was added to 50 mL of ethanol solution (10 mg / mL). The reaction was carried out at room temperature and 500 rpm in the dark for 24 h. The mixture was centrifuged and washed to obtain 30% DCA-modified R500 mesoporous silica. R500-30% PEG 1000 and R500-30% PEG1000 - The particle size and potential of DCA are as follows Figure 2 As shown, the surface potential gradually approaches electroneutrality from electronegativity. After DCA grafting, it becomes electronegative again, possibly because the carboxyl groups of DCA bind to the amino groups on the PEG surface, weakening the charge shielding effect of PEG itself. Characterization of the potential preliminarily confirms the successful modification with DCA. Figure 3 As shown, R500 exhibits the characteristic absorption peak of Si-O, at 1102 cm⁻¹. -1 and 801cm -1 R500-30% PEG 1000 and R500-30% PEG 1000 -DCA exhibits characteristic infrared absorption peaks for both PEG and DCA, including 2,958 cm⁻¹. -1 and 2,923cm -1 The CH extension vibration peak on the PEG at 1,460 cm⁻¹, and the peak at 1,460 cm⁻¹. -1 and 1,377cm -1 The CH bending vibration peak on the DCA at the specified location is consistent with the characteristic peak shapes of PEG and DCA, demonstrating the successful modification of DCA with PEG as a linker.

[0466] Example 76: 30% PEG with ester bonds 1000 DCA-modified R500 mesoporous silicon

[0467] S1: Mesoporous silica dispersion. 500 mg of the non-absorbent mesoporous silica support R500 prepared in Example 3 was dispersed in a mixture of 340 mL water and 200 mL ethanol, and the pH was adjusted to 4.0 with dilute hydrochloric acid.

[0468] S2: 30% PEG 1000 -OH modification. Add 30% silane-PEG to S1. 1000 -OH ethanol solution was added, and then ethanol solution was added to make the ethanol:water volume ratio of the reaction system 1:2. The reaction was carried out at 65℃ and 800rpm for 24h. The carrier was washed by centrifugation and resuspended in anhydrous ethanol to obtain 30% PEG. 1000 R500 solution modified with -OH.

[0469] S3: DCA carboxyl activation. 100 mg DCA was dissolved in 5 mL of LDM under a dry nitrogen atmosphere. 125 mg DCC was added, and the mixture was stirred at room temperature for 1 hour to form the activated intermediate O-acylurea.

[0470] S4: DCA-modified mesoporous silica. 5 mg of DPTS was added to the intermediate of S3, followed by the addition of 30% PEG from S2. 1000The modified mesoporous silica was stirred and reacted for 12-24 hours. The mixture was centrifuged and washed to obtain 30% DCA-modified R500 mesoporous silica.

[0471] Example 77: 30% PEG with thioether linkages 1000 DCA-modified R500 mesoporous silicon

[0472] S1: Mesoporous silica dispersion. 500 mg of the non-absorbent mesoporous silica support R500 prepared in Example 3 was dispersed in a mixture of 340 mL water and 200 mL ethanol, and the pH was adjusted to 4.0 with dilute hydrochloric acid.

[0473] S2: 30% PEG 1000 -Mal modification. Add 30% silane-PEG to S1. 1000 The ethanol solution was first prepared, and then ethanol was added to make the ethanol:water volume ratio of the reaction system 1:2. The reaction was carried out at 65°C and 800 rpm for 24 h. The carrier was washed by centrifugation and resuspended in DMF to obtain 30% PEG. 1000 -Mal-modified R500 solution.

[0474] S3: DCA-modified mesoporous silica. 100 mg of thiolized DCA was dissolved in 10 mL of LDM under a dry nitrogen atmosphere. 30% of the PEG from S2 was then added. 1000 The modified mesoporous silica was stirred and reacted for 12-24 hours. The mixture was centrifuged and washed to obtain 30% DCA-modified R500 mesoporous silica.

[0475] Example 78: 30% PEG with triazole linkage 1000 DCA-modified R500 mesoporous silicon

[0476] S1: Mesoporous silica dispersion. 500 mg of the non-absorbent mesoporous silica support R500 prepared in Example 3 was dispersed in a mixture of 340 mL water and 200 mL ethanol, and the pH was adjusted to 4.0 with dilute hydrochloric acid.

[0477] S2: 30% PEG 1000 -N3 modification. 30% silane-PEG was added to S1. 1000 -N3 ethanol solution was added, and then ethanol solution was added to make the ethanol:water volume ratio of the reaction system 1:2. The reaction was carried out at 65℃ and 800rpm for 24h. The carrier was washed by centrifugation and resuspended in DMF to obtain 30% PEG. 1000 -N3 modified R500 solution.

[0478] S3: DCA-modified mesoporous silica. Under a dry nitrogen atmosphere, 100 mg of alkynylated DCA, 5 mg of CuSO4·5H2O, and 10 mg of sodium ascorbate were dissolved in 5 mL of DMF. 30% of the PEG from S2 was then added. 1000 The modified mesoporous silica was reacted with stirring at room temperature for 12 h. The mixture was centrifuged and washed to obtain 30% DCA-modified R500 mesoporous silica.

[0479] Example 79: 30% PEG with carbamate bonds 1000 DCA-modified R500 mesoporous silicon

[0480] S1: Mesoporous silica dispersion. 500 mg of the non-absorbent mesoporous silica support R500 prepared in Example 3 was dispersed in a mixture of 340 mL water and 200 mL ethanol, and the pH was adjusted to 4.0 with dilute hydrochloric acid.

[0481] S2: 30% PEG 1000 -NCO modification. Add 30% silane-PEG to S1. 1000 -NCO ethanol solution was added, and then ethanol solution was added to make the ethanol:water volume ratio of the reaction system 1:2. The reaction was carried out at 65℃ and 800rpm for 24h. The carrier was washed by centrifugation and resuspended in DMF to obtain 30% PEG. 1000 -NCO modified R500 solution.

[0482] S3: DCA-modified mesoporous silica. 100 mg of isocyanated DCA was dissolved in 5 mL of LDM under a dry nitrogen atmosphere. 10 mg of DMAP was added as a catalyst, along with 30% PEG from S2. 1000 The modified mesoporous silica was reacted with stirring at room temperature for 6-12 h. The mixture was centrifuged and washed to obtain 30% DCA-modified R500 mesoporous silica.

[0483] Example 80: 10% PEG with amide bonds 2000 -DCA-modified R500 mesoporous silicon (R500-10% PEG) 2000 -DCA)

[0484] S1: Mesoporous silica dispersion. 500 mg of the non-absorbent mesoporous silica support R500 prepared in Example 3 was dispersed in a mixture of 340 mL water and 200 mL ethanol, and the pH was adjusted to 4.0 with dilute hydrochloric acid.

[0485] S2: 10% PEG 2000 -NH2 modification. 10% silane-PEG was added to S1. 2000The NH2 ethanol solution was added, and then ethanol solution was added to make the ethanol:water volume ratio of the reaction system 1:2. The reaction was carried out at 65℃ and 800rpm for 24h. The carrier was washed by centrifugation and resuspended in anhydrous ethanol to obtain 10% PEG. 2000 -NH2 modified R500 solution.

[0486] S3: DCA carboxyl activation. Dissolve 109.8 mg DCA in 11.0 mL DMF. Then add 21 mg NHS and 56 mg EDC sequentially, activate at room temperature for 15 min, and adjust pH to 5.8.

[0487] S4: DCA-modified mesoporous silica. The activation solution of S3 was added to 50 mL of ethanol solution (10 mg / mL). The reaction was carried out at room temperature and 500 rpm in the dark for 24 h. The mixture was centrifuged and washed to obtain 10% DCA-modified R500 mesoporous silica. R500-10% PEG 2000 and R500-10% PEG 2000 - The particle size and potential of DCA are as follows Figure 2 As shown, the surface potential gradually approaches electroneutrality from electronegativity, and then becomes electronegative again after DCA grafting. This may be because the carboxyl groups of DCA bind to the amino groups on the PEG surface, weakening the charge shielding effect of PEG itself. Characterization of the potential demonstrates the successful modification with DCA.

[0488] Example 81: 10% PEG with ester bonds 2000 DCA-modified R500 mesoporous silicon

[0489] S1: Mesoporous silica dispersion. 500 mg of the non-absorbent mesoporous silica support R500 prepared in Example 3 was dispersed in a mixture of 340 mL water and 200 mL ethanol, and the pH was adjusted to 4.0 with dilute hydrochloric acid.

[0490] S2: 10% PEG 2000 -OH modification. Add 10% silane-PEG to S1. 2000 -OH ethanol solution was added, and then ethanol solution was added to make the ethanol:water volume ratio of the reaction system 1:2. The reaction was carried out at 65℃ and 800rpm for 24h. The carrier was washed by centrifugation and resuspended in anhydrous ethanol to obtain 10% PEG. 2000 R500 solution modified with -OH.

[0491] S3: DCA carboxyl activation. Under a dry nitrogen atmosphere, 100 mg DCA was dissolved in 5 mL DMF. 125 mg DCC was added, and the mixture was stirred at room temperature for 1 h to form the activated intermediate O-acylurea.

[0492] S4: DCA-modified mesoporous silica. 5 mg of DPTS was added to the intermediate from S3, followed by the addition of 10% PEG from S2. 2000 The modified mesoporous silica was stirred and reacted for 12-24 hours. The mixture was centrifuged and washed to obtain 10% DCA-modified R500 mesoporous silica.

[0493] Example 82, 10% PEG with thioether linkage 2000 DCA-modified R500 mesoporous silicon

[0494] S1: Mesoporous silica dispersion. 500 mg of the non-absorbent mesoporous silica support R500 prepared in Example 3 was dispersed in a mixture of 340 mL water and 200 mL ethanol, and the pH was adjusted to 4.0 with dilute hydrochloric acid.

[0495] S2: 10% PEG 2000 -Mal modification. Add 10% silane-PEG to S1. 2000 The ethanol solution was first prepared, and then ethanol was added to make the ethanol:water volume ratio of the reaction system 1:2. The reaction was carried out at 65°C and 800 rpm for 24 h. The carrier was washed by centrifugation and resuspended in DMF to obtain 10% PEG. 2000 -Mal-modified R500 solution.

[0496] S3: DCA-modified mesoporous silica. 100 mg of thiolized DCA was dissolved in 10 mL of LDM under a dry nitrogen atmosphere. 10% PEG from S2 was then added. 2000 The modified mesoporous silica was stirred and reacted for 12-24 hours. The mixture was centrifuged and washed to obtain 10% DCA-modified R500 mesoporous silica.

[0497] Example 83: 10% PEG with triazole linkage 2000 DCA-modified R500 mesoporous silicon

[0498] S1: Mesoporous silica dispersion. 500 mg of the non-absorbent mesoporous silica support R500 prepared in Example 3 was dispersed in a mixture of 340 mL water and 200 mL ethanol, and the pH was adjusted to 4.0 with dilute hydrochloric acid.

[0499] S2: 10% PEG 2000 -N3 modification. 10% silane-PEG was added to S1. 2000 -N3 ethanol solution was added, and then ethanol solution was added to make the ethanol:water volume ratio of the reaction system 1:2. The reaction was carried out at 65℃ and 800rpm for 24h. The carrier was washed by centrifugation and resuspended in DMF to obtain 10% PEG. 2000 -N3 modified R500 solution.

[0500] S3: DCA-modified mesoporous silica. Under a dry nitrogen atmosphere, 100 mg of alkynylated DCA, 5 mg of CuSO4·5H2O, and 10 mg of sodium ascorbate were dissolved in 5 mL of DMF. 10% PEG from S2 was then added. 2000 The modified mesoporous silica was reacted with stirring at room temperature for 12 h. The mixture was centrifuged and washed to obtain 10% DCA-modified R500 mesoporous silica.

[0501] Example 84: 10% PEG with carbamate linkages 2000 DCA-modified R500 mesoporous silicon

[0502] S1: Mesoporous silica dispersion. 500 mg of the non-absorbent mesoporous silica support R500 prepared in Example 3 was dispersed in a mixture of 340 mL water and 200 mL ethanol, and the pH was adjusted to 4.0 with dilute hydrochloric acid.

[0503] S2: 10% PEG 2000 -NCO modification. Add 10% silane-PEG to S1. 2000 -NCO ethanol solution was added, and then ethanol solution was added to make the ethanol:water volume ratio of the reaction system 1:2. The reaction was carried out at 65℃ and 800rpm for 24h. The carrier was washed by centrifugation and resuspended in DMF to obtain 10% PEG. 2000 -NCO modified R500 solution.

[0504] S3: DCA-modified mesoporous silica. 100 mg of isocyanated DCA was dissolved in 5 mL of LDM under a dry nitrogen atmosphere. 10 mg of DMAP was added as a catalyst, along with 10% PEG from S2. 2000 The modified mesoporous silica was reacted with stirring at room temperature for 6-12 h. The mixture was centrifuged and washed to obtain 10% DCA-modified R500 mesoporous silica.

[0505] Example 85: 20% PEG with amide bonds 2000 -DCA-modified R500 mesoporous silica (R500-20% PEG) 2000 -DCA)

[0506] S1: Mesoporous silica dispersion. 500 mg of the non-absorbent mesoporous silica support R500 prepared in Example 3 was dispersed in a mixture of 340 mL water and 200 mL ethanol, and the pH was adjusted to 4.0 with dilute hydrochloric acid.

[0507] S2: 20% PEG 2000 -NH2 modification. 20% silane-PEG was added to S1. 2000The NH2 ethanol solution was added, and then ethanol solution was added to make the ethanol:water volume ratio of the reaction system 1:2. The reaction was carried out at 65℃ and 800rpm for 24h. The carrier was washed by centrifugation and resuspended in anhydrous ethanol to obtain 20% PEG. 2000 -NH2 modified R500 solution.

[0508] S3: DCA carboxyl activation. Dissolve 109.8 mg DCA in 11.0 mL DMF. Then add 21 mg NHS and 56 mg EDC sequentially, activate at room temperature for 15 min, and adjust pH to 5.8.

[0509] S4: DCA-modified mesoporous silica. The activation solution of S3 was added to 50 mL of ethanol solution (10 mg / mL). The reaction was carried out at room temperature and 500 rpm in the dark for 24 h. The mixture was centrifuged and washed to obtain 20% DCA-modified R500 mesoporous silica. R500-20% PEG 2000 and R500-20% PEG 2000 - The particle size and potential of DCA are as follows Figure 2 As shown, the surface potential gradually approaches electroneutrality from electronegativity, and then becomes electronegative again after DCA grafting. This may be because the carboxyl groups of DCA bind to the amino groups on the PEG surface, weakening the charge shielding effect of PEG itself. Characterization of the potential demonstrates the successful modification with DCA.

[0510] Example 86: 20% PEG with ester bonds 2000 DCA-modified R500 mesoporous silicon

[0511] S1: Mesoporous silica dispersion. 500 mg of the non-absorbent mesoporous silica support R500 prepared in Example 3 was dispersed in a mixture of 340 mL water and 200 mL ethanol, and the pH was adjusted to 4.0 with dilute hydrochloric acid.

[0512] S2: 20% PEG 2000 -OH modification. Add 20% silane-PEG to S1. 2000 -OH ethanol solution was added, and then ethanol solution was added to make the ethanol:water volume ratio of the reaction system 1:2. The reaction was carried out at 65℃ and 800rpm for 24h. The carrier was washed by centrifugation and resuspended in anhydrous ethanol to obtain 20% PEG. 2000 R500 solution modified with -OH.

[0513] S3: DCA carboxyl activation. Under a dry nitrogen atmosphere, 100 mg DCA was dissolved in 5 mL DMF. 125 mg DCC was added, and the mixture was stirred at room temperature for 1 h to form the activated intermediate O-acylurea.

[0514] S4: DCA-modified mesoporous silica. The intermediate from S3 was added to 5 mg of DPTS, followed by the addition of 20% PEG from S2. 2000 The modified mesoporous silica was stirred and reacted for 12-24 hours. The mixture was centrifuged and washed to obtain 20% DCA-modified R500 mesoporous silica.

[0515] Example 87: 20% PEG with thioether linkages 2000 DCA-modified R500 mesoporous silicon

[0516] S1: Mesoporous silica dispersion. 500 mg of the non-absorbent mesoporous silica support R500 prepared in Example 3 was dispersed in a mixture of 340 mL water and 200 mL ethanol, and the pH was adjusted to 4.0 with dilute hydrochloric acid.

[0517] S2: 20% PEG 2000 -Mal modification. Add 20% silane-PEG to S1. 2000 The ethanol solution was first prepared, and then ethanol was added to make the ethanol:water volume ratio of the reaction system 1:2. The reaction was carried out at 65°C and 800 rpm for 24 h. The carrier was washed by centrifugation and resuspended in DMF to obtain 20% PEG. 2000 -Mal-modified R500 solution.

[0518] S3: DCA-modified mesoporous silica. 100 mg of thiolized DCA was dissolved in 10 mL of LDM under a dry nitrogen atmosphere. 20% PEG from S2 was then added. 2000 The modified mesoporous silica was stirred and reacted for 12-24 hours. The mixture was centrifuged and washed to obtain 20% DCA-modified R500 mesoporous silica.

[0519] Example 88: 20% PEG with triazole linkage 2000 DCA-modified R500 mesoporous silicon

[0520] S1: Mesoporous silica dispersion. 500 mg of the non-absorbent mesoporous silica support R500 prepared in Example 3 was dispersed in a mixture of 340 mL water and 200 mL ethanol, and the pH was adjusted to 4.0 with dilute hydrochloric acid.

[0521] S2: 20% PEG 2000 -N3 modification. 20% silane-PEG was added to S1. 2000 -N3 ethanol solution was added, and then ethanol solution was added to make the ethanol:water volume ratio of the reaction system 1:2. The reaction was carried out at 65℃ and 800rpm for 24h. The carrier was washed by centrifugation and resuspended in DMF to obtain 20% PEG. 2000 -N3 modified R500 solution.

[0522] S3: DCA-modified mesoporous silica. Under a dry nitrogen atmosphere, 100 mg of alkynylated DCA, 5 mg of CuSO4·5H2O, and 10 mg of sodium ascorbate were dissolved in 5 mL of DMF. 20% of the PEG from S2 was then added. 2000 The modified mesoporous silica was reacted with stirring at room temperature for 12 h. The mixture was centrifuged and washed to obtain 20% DCA-modified R500 mesoporous silica.

[0523] Example 89: 20% PEG with carbamate bonds 2000 DCA-modified R500 mesoporous silicon

[0524] S1: Mesoporous silica dispersion. 500 mg of the non-absorbent mesoporous silica support R500 prepared in Example 3 was dispersed in a mixture of 340 mL water and 200 mL ethanol, and the pH was adjusted to 4.0 with dilute hydrochloric acid.

[0525] S2: 20% PEG 2000 -NCO modification. Add 20% silane-PEG to S1. 2000 -NCO ethanol solution was added, and then ethanol solution was added to make the ethanol:water volume ratio of the reaction system 1:2. The reaction was carried out at 65℃ and 800rpm for 24h. The carrier was washed by centrifugation and resuspended in DMF to obtain 20% PEG. 2000 -NCO modified R500 solution.

[0526] S3: DCA-modified mesoporous silica. 100 mg of isocyanated DCA was dissolved in 5 mL of LDM under a dry nitrogen atmosphere. 10 mg of DMAP was added as a catalyst, along with 20% PEG from S2. 2000 The modified mesoporous silica was reacted with stirring at room temperature for 6-12 h. The mixture was centrifuged and washed to obtain 20% DCA-modified R500 mesoporous silica.

[0527] Example 90: 30% PEG with amide bonds 2000 -DCA-modified R500 mesoporous silicon (R500-30% PEG) 2000 -DCA)

[0528] S1: Mesoporous silica dispersion. 500 mg of the non-absorbent mesoporous silica support R500 prepared in Example 3 was dispersed in a mixture of 340 mL water and 200 mL ethanol, and the pH was adjusted to 4.0 with dilute hydrochloric acid.

[0529] S2: 30% PEG 2000 -NH2 modification. 30% silane-PEG was added to S1. 2000-NH2 ethanol solution was added, and then ethanol solution was added to make the ethanol:water volume ratio of the reaction system 1:2. The reaction was carried out at 65℃ and 800rpm for 24h. The carrier was washed by centrifugation and resuspended in anhydrous ethanol to obtain 30% PEG. 2000 -NH2 modified R500 solution.

[0530] S3: DCA carboxyl activation. Dissolve 109.8 mg DCA in 11.0 mL DMF. Then add 21 mg NHS and 56 mg EDC sequentially, activate at room temperature for 15 min, and adjust pH to 5.8.

[0531] S4: DCA-modified mesoporous silica. The activation solution of S3 was added to 50 mL of ethanol solution (10 mg / mL). The reaction was carried out at room temperature and 500 rpm in the dark for 24 h. The mixture was centrifuged and washed to obtain 30% DCA-modified R500 mesoporous silica. R500-30% PEG 2000 and R500-30% PEG 2000 - The particle size and potential of DCA are as follows Figure 2 As shown, the surface potential gradually approaches electroneutrality from electronegativity, and then becomes electronegative again after DCA grafting. This may be because the carboxyl groups of DCA bind to the amino groups on the PEG surface, weakening the charge shielding effect of PEG itself. Characterization of the potential demonstrates the successful modification with DCA.

[0532] Example 91: 30% PEG with ester bonds 2000 DCA-modified R500 mesoporous silicon

[0533] S1: Mesoporous silica dispersion. 500 mg of the non-absorbent mesoporous silica support R500 prepared in Example 3 was dispersed in a mixture of 340 mL water and 200 mL ethanol, and the pH was adjusted to 4.0 with dilute hydrochloric acid.

[0534] S2: 30% PEG 2000 -OH modification. Add 30% silane-PEG to S1. 2000 -OH ethanol solution was added, and then ethanol solution was added to make the ethanol:water volume ratio of the reaction system 1:2. The reaction was carried out at 65℃ and 800rpm for 24h. The carrier was washed by centrifugation and resuspended in anhydrous ethanol to obtain 30% PEG. 2000 R500 solution modified with -OH.

[0535] S3: DCA carboxyl activation. 100 mg DCA was dissolved in 5 mL of LDM under a dry nitrogen atmosphere. 125 mg DCC was added, and the mixture was stirred at room temperature for 1 hour to form the activated intermediate O-acylurea.

[0536] S4: DCA-modified mesoporous silica. 5 mg of DPTS was added to the intermediate of S3, followed by the addition of 30% PEG from S2. 2000 The modified mesoporous silica was stirred and reacted for 12-24 hours. The mixture was centrifuged and washed to obtain 30% DCA-modified R500 mesoporous silica.

[0537] Example 92, 30% PEG with thioether linkage 2000 DCA-modified R500 mesoporous silicon

[0538] S1: Mesoporous silica dispersion. 500 mg of the non-absorbent mesoporous silica support R500 prepared in Example 3 was dispersed in a mixture of 340 mL water and 200 mL ethanol, and the pH was adjusted to 4.0 with dilute hydrochloric acid.

[0539] S2: 30% PEG 2000 -Mal modification. Add 30% silane-PEG to S1. 2000 The ethanol solution was first prepared, and then ethanol was added to make the ethanol:water volume ratio of the reaction system 1:2. The reaction was carried out at 65°C and 800 rpm for 24 h. The carrier was washed by centrifugation and resuspended in DMF to obtain 30% PEG. 2000 -Mal-modified R500 solution.

[0540] S3: DCA-modified mesoporous silica. 100 mg of thiolized DCA was dissolved in 10 mL of LDM under a dry nitrogen atmosphere. 30% of the PEG from S2 was then added. 2000 The modified mesoporous silica was stirred and reacted for 12-24 hours. The mixture was centrifuged and washed to obtain 30% DCA-modified R500 mesoporous silica.

[0541] Example 93: 30% PEG with triazole linkage 2000 DCA-modified R500 mesoporous silicon

[0542] S1: Mesoporous silica dispersion. 500 mg of the non-absorbent mesoporous silica support R500 prepared in Example 3 was dispersed in a mixture of 340 mL water and 200 mL ethanol, and the pH was adjusted to 4.0 with dilute hydrochloric acid.

[0543] S2: 30% PEG 2000 -N3 modification. 30% silane-PEG was added to S1. 2000 -N3 ethanol solution was added, and then ethanol solution was added to make the ethanol:water volume ratio of the reaction system 1:2. The reaction was carried out at 65℃ and 800rpm for 24h. The carrier was washed by centrifugation and resuspended in DMF to obtain 30% PEG. 2000 -N3 modified R500 solution.

[0544] S3: DCA-modified mesoporous silica. Under a dry nitrogen atmosphere, 100 mg of alkynylated DCA, 5 mg of CuSO4·5H2O, and 10 mg of sodium ascorbate were dissolved in 5 mL of DMF. 30% of the PEG from S2 was then added. 2000 The modified mesoporous silica was reacted with stirring at room temperature for 12 h. The mixture was centrifuged and washed to obtain 30% DCA-modified R500 mesoporous silica.

[0545] Example 94: 30% PEG with carbamate bonds 2000 DCA-modified R500 mesoporous silicon

[0546] S1: Mesoporous silica dispersion. 500 mg of the non-absorbent mesoporous silica support R500 prepared in Example 3 was dispersed in a mixture of 340 mL water and 200 mL ethanol, and the pH was adjusted to 4.0 with dilute hydrochloric acid.

[0547] S2: 30% PEG 2000 -NCO modification. Add 30% silane-PEG to S1. 2000 -NCO ethanol solution was added, and then ethanol solution was added to make the ethanol:water volume ratio of the reaction system 1:2. The reaction was carried out at 65℃ and 800rpm for 24h. The carrier was washed by centrifugation and resuspended in DMF to obtain 30% PEG. 2000 -NCO modified R500 solution.

[0548] S3: DCA-modified mesoporous silica. 100 mg of isocyanated DCA was dissolved in 5 mL of LDM under a dry nitrogen atmosphere. 10 mg of DMAP was added as a catalyst, along with 30% PEG from S2. 2000 The modified mesoporous silica was reacted with stirring at room temperature for 6-12 h. The mixture was centrifuged and washed to obtain 30% DCA-modified R500 mesoporous silica.

[0549] Example 95: 10% PEG with amide bonds 1000 -DCA-modified R1000PLGA carrier (R1000-10% PEG) 1000 -DCA)

[0550] S1: DCA carboxyl activation. 100 mg DCA was dissolved in 5 mL of LDM under a dry nitrogen atmosphere. Then, 21 mg NHS and 56 mg EDC were added sequentially, and the mixture was activated at room temperature for 15 min, adjusting the pH to 5.8.

[0551] S2: BOC-PEG 1000 Preparation of DCA. An equal mass of NH2-PEG was added to the intermediate of S1.1000 -BOC. The reaction was carried out at room temperature and 500 rpm in the dark for 24 h. After purification, BOC-PEG was obtained. 1000 -DCA.

[0552] S3: NH2-PEG 1000 Preparation of DCA. BOC-PEG obtained in S2 was treated with a 50% TFA / DCM solution. 1000 -DCA was used to remove the BOC protecting group. Subsequently, NH2-PEG was obtained by precipitation and purification with diethyl ether. 1000 -DCA.

[0553] S4: PLGA carrier carboxyl activation. 500 mg of the non-absorbable poly(lactic-co-glycolic acid) carrier R1000 prepared in Example 4 was dispersed in 500 mL of water. Then, 21 mg of NHS and 56 mg of EDC were added sequentially, and the mixture was activated at room temperature for 15 min, and the pH was adjusted to 5.8.

[0554] S5: DCA-modified PLGA support. NH2-PEG prepared in S3 with a 10% PLGA mass ratio. 1000 DCA was added to the activation solution of S4. The reaction was carried out at room temperature and 500 rpm in the dark for 24 h. The mixture was centrifuged and washed to obtain the 10% DCA-modified R1000PLGA support. Results are as follows... Figure 4 As shown, R1000-10% PEG 1000 -DCA appears as a rod-shaped structure under an electron microscope, with a length and width of approximately 2000 nm and 400 nm, respectively. R1000-10% PEG 1000 and R1000-10% PEG 1000 - The particle size and potential of DCA are as follows Figure 2 As shown, the surface potential gradually approaches electroneutrality from electronegativity, and then becomes electronegative again after DCA grafting. This may be because the carboxyl groups of DCA bind to the amino groups on the PEG surface, weakening the charge shielding effect of PEG itself. Characterization of the potential demonstrates the successful modification with DCA.

[0555] Example 96: 20% PEG with amide bonds 1000 -DCA-modified R1000PLGA carrier (R1000-20% PEG) 1000 -DCA)

[0556] S1: DCA carboxyl activation. Under a dry nitrogen atmosphere, 100 mg DCA was dissolved in 5 mL DMF. Then, 21 mg NHS and 56 mg EDC were added sequentially, and the mixture was activated at room temperature for 15 min, adjusting the pH to 5.8.

[0557] Preparation of S2: BOC-PEG1000-DCA. An equal mass of NH2-PEG1000-BOC was added to the intermediate in S1. The reaction was carried out at room temperature and 500 rpm in the dark for 24 h. After purification, BOC-PEG1000-DCA was obtained.

[0558] S3: Preparation of NH2-PEG1000-DCA. The BOC-PEG1000-DCA obtained in S2 was treated with 50% TFA / DCM solution to remove the BOC protecting groups. Subsequently, it was precipitated with diethyl ether and purified to obtain NH2-PEG1000-DCA.

[0559] S4: PLGA carrier carboxyl activation. 500 mg of the R1000PLGA carrier prepared in Example 4 was dispersed in 500 mL of water. Then, 21 mg of NHS and 56 mg of EDC were added sequentially, and the mixture was activated at room temperature for 15 min, and the pH was adjusted to 5.8.

[0560] S5: DCA-modified PLGA support. NH2-PEG1000-DCA (20% PLGA by mass ratio) prepared in S3 was added to the activation solution in S4. The reaction was carried out at room temperature and 500 rpm in the dark for 24 h. The mixture was centrifuged and washed to obtain the 20% DCA-modified R1000PLGA support. R1000-20% PEG 1000 and R1000-20% PEG 1000 - The particle size and potential of DCA are as follows Figure 2 As shown, the surface potential gradually approaches electroneutrality from electronegativity, and then becomes electronegative again after DCA grafting. This may be because the carboxyl groups of DCA bind to the amino groups on the PEG surface, weakening the charge shielding effect of PEG itself. Characterization of the potential demonstrates the successful modification with DCA.

[0561] Example 97: 30% PEG with amide bonds 1000 -DCA-modified R1000PLGA carrier (R1000-30% PEG) 1000 -DCA)

[0562] S1: DCA carboxyl activation. Under a dry nitrogen atmosphere, 100 mg DCA was dissolved in 5 mL DMF. Then, 21 mg NHS and 56 mg EDC were added sequentially, and the mixture was activated at room temperature for 15 min, adjusting the pH to 5.8.

[0563] S2: BOC-PEG 1000 Preparation of DCA. An equal mass of NH2-PEG was added to the intermediate of S1. 1000-BOC. The reaction was carried out at room temperature and 500 rpm in the dark for 24 h. After purification, BOC-PEG was obtained. 1000 -DCA.

[0564] S3: NH2-PEG 1000 Preparation of DCA. BOC-PEG obtained in S2 was treated with a 50% TFA / DCM solution. 1000 -DCA was used to remove the BOC protecting group. Subsequently, NH2-PEG was obtained by precipitation and purification with diethyl ether. 1000 -DCA.

[0565] S4: PLGA carrier carboxyl activation. 500 mg of the R1000PLGA carrier prepared in Example 4 was dispersed in 500 mL of water. Then, 21 mg of NHS and 56 mg of EDC were added sequentially, and the mixture was activated at room temperature for 15 min, and the pH was adjusted to 5.8.

[0566] S5: DCA-modified PLGA support. NH2-PEG prepared in S3 with a 30% PLGA mass ratio. 1000 DCA was added to the activation solution of S4. The reaction was carried out at room temperature and 500 rpm in the dark for 24 hours. The mixture was centrifuged and washed to obtain a 30% DCA-modified R1000PLGA support. R1000-30% PEG 1000 and R1000-30% PEG 1000 - The particle size and potential of DCA are as follows Figure 2 As shown, the surface potential gradually approaches electroneutrality from electronegativity, and then becomes electronegative again after DCA grafting. This may be because the carboxyl groups of DCA bind to the amino groups on the PEG surface, weakening the charge shielding effect of PEG itself. Characterization of the potential demonstrates the successful modification with DCA.

[0567] Example 98, 10% PEG with amide bonds 2000 -DCA-modified R1000PLGA carrier (R1000-10% PEG) 2000 -DCA)

[0568] S1: DCA carboxyl activation. Under a dry nitrogen atmosphere, 100 mg DCA was dissolved in 5 mL DMF. Then, 21 mg NHS and 56 mg EDC were added sequentially, and the mixture was activated at room temperature for 15 min, adjusting the pH to 5.8.

[0569] S2: BOC-PEG 2000 Preparation of DCA. An equal mass of NH2-PEG was added to the intermediate of S1. 2000-BOC. The reaction was carried out at room temperature and 500 rpm in the dark for 24 h. After purification, BOC-PEG was obtained. 2000 -DCA.

[0570] S3: NH2-PEG 2000 Preparation of -DCA. BOC-PEG2000-DCA obtained in S2 was treated with 50% TFA / DCM solution to remove the BOC protecting groups. Subsequently, it was precipitated with diethyl ether and purified to obtain NH2-PEG. 2000 -DCA.

[0571] S4: PLGA carrier carboxyl activation. 500 mg of the R1000PLGA carrier prepared in Example 4 was dispersed in 500 mL of water. Then, 21 mg of NHS and 56 mg of EDC were added sequentially, and the mixture was activated at room temperature for 15 min, and the pH was adjusted to 5.8.

[0572] S5: DCA-modified PLGA support. NH2-PEG prepared in S3 with a 10% PLGA mass ratio. 2000 DCA was added to the activation solution of S4. The reaction was carried out at room temperature and 500 rpm in the dark for 24 hours. The mixture was centrifuged and washed to obtain a 10% DCA-modified R1000PLGA support. R1000-10% PEG 2000 and R1000-10% PEG 2000 - The particle size and potential of DCA are as follows Figure 2 As shown, the surface potential gradually approaches electroneutrality from electronegativity, and then becomes electronegative again after DCA grafting. This may be because the carboxyl groups of DCA bind to the amino groups on the PEG surface, weakening the charge shielding effect of PEG itself. Characterization of the potential demonstrates the successful modification with DCA.

[0573] Example 99: 20% PEG with amide bonds 2000 -DCA-modified R1000PLGA carrier (R1000-20% PEG) 2000 -DCA)

[0574] S1: DCA carboxyl activation. 100 mg DCA was dissolved in 5 mL of LDM under a dry nitrogen atmosphere. Then, 21 mg NHS and 56 mg EDC were added sequentially, and the mixture was activated at room temperature for 15 min, adjusting the pH to 5.8.

[0575] Preparation of S2: BOC-PEG2000-DCA. An equal mass of NH2-PEG was added to the intermediate from S1. 2000 -BOC. The reaction was carried out at room temperature and 500 rpm in the dark for 24 h. After purification, BOC-PEG was obtained. 2000 -DCA.

[0576] S3: NH2-PEG 2000 Preparation of DCA. BOC-PEG obtained in S2 was treated with a 50% TFA / DCM solution. 2000 -DCA was used to remove the BOC protecting group. Subsequently, NH2-PEG was obtained by precipitation and purification with diethyl ether. 2000 -DCA.

[0577] S4: PLGA carrier carboxyl activation. 500 mg of the R1000PLGA carrier prepared in Example 4 was dispersed in 500 mL of water. Then, 21 mg of NHS and 56 mg of EDC were added sequentially, and the mixture was activated at room temperature for 15 min, and the pH was adjusted to 5.8.

[0578] S5: DCA-modified PLGA support. NH2-PEG prepared in S3 with a 20% PLGA mass ratio. 2000 DCA was added to the activation solution of S4. The reaction was carried out at room temperature and 500 rpm in the dark for 24 hours. The mixture was centrifuged and washed to obtain a 20% DCA-modified R1000PLGA support. R1000-20% PEG 2000 and R1000-20% PEG 2000 - The particle size and potential of DCA are as follows Figure 2 As shown, the surface potential gradually approaches electroneutrality from electronegativity, and then becomes electronegative again after DCA grafting. This may be because the carboxyl groups of DCA bind to the amino groups on the PEG surface, weakening the charge shielding effect of PEG itself. Characterization of the potential demonstrates the successful modification with DCA.

[0579] Example 100: 30% PEG with amide bonds 2000 -DCA-modified R1000PLGA carrier (R1000-30% PEG) 2000 -DCA)

[0580] S1: DCA carboxyl activation. Under a dry nitrogen atmosphere, 100 mg DCA was dissolved in 5 mL DMF. Then, 21 mg NHS and 56 mg EDC were added sequentially, and the mixture was activated at room temperature for 15 min, adjusting the pH to 5.8.

[0581] S2: BOC-PEG 2000 Preparation of DCA. An equal mass of NH2-PEG was added to the intermediate of S1. 2000 -BOC. The reaction was carried out at room temperature and 500 rpm in the dark for 24 h. After purification, BOC-PEG was obtained. 2000 -DCA.

[0582] S3: NH2-PEG2000 Preparation of DCA. BOC-PEG obtained in S2 was treated with a 50% TFA / DCM solution. 2000 -DCA was used to remove the BOC protecting group. Subsequently, NH2-PEG was obtained by precipitation and purification with diethyl ether. 2000 -DCA.

[0583] S4: PLGA carrier carboxyl activation. 500 mg of the R1000PLGA carrier prepared in Example 4 was dispersed in 500 mL of water. Then, 21 mg of NHS and 56 mg of EDC were added sequentially, and the mixture was activated at room temperature for 15 min, and the pH was adjusted to 5.8.

[0584] S5: DCA-modified PLGA support. NH2-PEG prepared in S3 with a 30% PLGA mass ratio. 2000 DCA was added to the activation solution of S4. The reaction was carried out at room temperature and 500 rpm in the dark for 24 hours. The mixture was centrifuged and washed to obtain a 30% DCA-modified R1000PLGA support. R1000-30% PEG 2000 and R1000-30% PEG 2000 - The particle size and potential of DCA are as follows Figure 2 As shown, the surface potential gradually approaches electroneutrality from electronegativity, and then becomes electronegative again after DCA grafting. This may be because the carboxyl groups of DCA bind to the amino groups on the PEG surface, weakening the charge shielding effect of PEG itself. Characterization of the potential demonstrates the successful modification with DCA.

[0585] Example 101: 10% PEG with ester bonds 1000 -DCA modified R1000PLGA carrier

[0586] S1: DCA carboxyl activation. Under a dry nitrogen atmosphere, 100 mg DCA was dissolved in 5 mL DMF. 125 mg DCC was added, and the mixture was stirred at room temperature for 1 h to form the activated intermediate O-acylurea.

[0587] S2: NH2-PEG 1000 Preparation of DCA: 5 mg of DPTS was added to the intermediate of S1, followed by an equal mass of NH2-PEG. 1000 -OH, stir the reaction for 12-24 h. Filter, wash and freeze-dry the mixture to obtain NH2-PEG. 1000 -DCA.

[0588] S3: PLGA carrier carboxyl activation. 500 mg of the non-absorbable poly(lactic-co-glycolic acid) carrier R1000 prepared in Example 4 was dispersed in 500 mL of water. Then, 21 mg of NHS and 56 mg of EDC were added sequentially, and the mixture was activated at room temperature for 15 min, and the pH was adjusted to 5.8.

[0589] S4: DCA-modified PLGA support. 10% PLGA prepared in S2 was then used in NH2-PEG. 1000 DCA was added to the activation solution of S3. The reaction was carried out at room temperature and 500 rpm in the dark for 24 h. The mixture was centrifuged and washed to obtain the R1000PLGA support modified with 10% DCA.

[0590] Example 102: 10% PEG with thioether linkage 1000 DCA-modified R1000 mesoporous silicon

[0591] S1: NH2-PEG 1000 Preparation of DCA: Under a dry nitrogen atmosphere, 100 mg of thiolized DCA was dissolved in 10 mL of DMF. An equal mass of NH2-PEG was then added. 1000 -Mal, stir the reaction for 12-24 h. Filter, wash and freeze-dry the mixture to obtain NH2-PEG. 1000 -DCA.

[0592] S2: PLGA carboxyl activation. 500 mg of the non-absorbable poly(lactic-co-glycolic acid) carrier R1000 prepared in Example 4 was dispersed in 500 mL of water. Then, 21 mg of NHS and 56 mg of EDC were added sequentially, and the mixture was activated at room temperature for 15 min, and the pH was adjusted to 5.8.

[0593] S3: DCA-modified PLGA support. 10% PLGA prepared in S1 was then used in NH2-PEG. 1000 DCA was added to the activation solution of S2. The reaction was carried out at room temperature and 500 rpm in the dark for 24 h. The mixture was centrifuged and washed to obtain the R1000PLGA support modified with 10% DCA.

[0594] Example 103: 10% PEG with triazole linkage 1000 DCA-modified R1000 mesoporous silicon

[0595] S1: NH2-PEG 1000 Preparation of DCA: Under a dry nitrogen atmosphere, 100 mg of alkynylated DCA, 5 mg of CuSO4·5H2O, and 10 mg of sodium ascorbate were dissolved in 5 mL of DMF. An equal mass of NH2-PEG was then added. 1000The reaction was carried out under N3 at room temperature with stirring for 12 h. The mixture was filtered, washed, and lyophilized to obtain NH2-PEG. 1000 -DCA.

[0596] S2: PLGA carboxyl activation. 500 mg of the non-absorbable poly(lactic-co-glycolic acid) carrier R1000 prepared in Example 4 was dispersed in 500 mL of water. Then, 21 mg of NHS and 56 mg of EDC were added sequentially, and the mixture was activated at room temperature for 15 min, and the pH was adjusted to 5.8.

[0597] S3: DCA-modified PLGA support. 10% PLGA prepared in S1 was then used in NH2-PEG. 1000 DCA was added to the activation solution of S2. The reaction was carried out at room temperature and 500 rpm in the dark for 24 h. The mixture was centrifuged and washed to obtain the R1000PLGA support modified with 10% DCA.

[0598] Example 104: 10% PEG with carbamate linkages 1000 DCA-modified R1000 mesoporous silicon

[0599] S1: NH2-PEG 1000 Preparation of DCA: Under a dry nitrogen atmosphere, 100 mg of isocyanated DCA was dissolved in 5 mL of DMF. 10 mg of DMAP was added as a catalyst, and an equal mass of NH2-PEG was added. 1000 -NCO was reacted at room temperature with stirring for 6-12 h. The mixture was then filtered, washed, and lyophilized to obtain NH2-PEG. 1000 -DCA.

[0600] S2: PLGA carboxyl activation. 500 mg of the non-absorbable poly(lactic-co-glycolic acid) carrier R1000 prepared in Example 4 was dispersed in 500 mL of water. Then, 21 mg of NHS and 56 mg of EDC were added sequentially, and the mixture was activated at room temperature for 15 min, and the pH was adjusted to 5.8.

[0601] S3: DCA-modified PLGA support. 10% PLGA prepared in S1 was then used in NH2-PEG. 1000 DCA was added to the activation solution of S2. The reaction was carried out at room temperature and 500 rpm in the dark for 24 h. The mixture was centrifuged and washed to obtain the R1000PLGA support modified with 10% DCA.

[0602] Example 105: 20% PEG with amide bonds 1000 -LCA-modified R500 mesoporous silica (R500-20% PEG) 1000 -LCA)

[0603] S1: Mesoporous silica dispersion. 500 mg of the non-absorbent mesoporous silica support R500 prepared in Example 3 was dispersed in a mixture of 340 mL water and 200 mL ethanol, and the pH was adjusted to 4.0 with dilute hydrochloric acid.

[0604] S2: 20% PEG 1000 -NH2 modification. 20% silane-PEG was added to S1. 1000 The NH2 ethanol solution was added, and then ethanol solution was added to make the ethanol:water volume ratio of the reaction system 1:2. The reaction was carried out at 65℃ and 800rpm for 24h. The carrier was washed by centrifugation and resuspended in anhydrous ethanol to obtain 20% PEG. 1000 -NH2 modified R500 solution.

[0605] S3: Lithocholic acid (LCA) carboxyl activation. 105.3 mg DCA was dissolved in 10.6 mL LDM. Then 20.2 mg NHS and 53.7 mg EDC were added sequentially, and the mixture was activated at room temperature for 15 min.

[0606] S4: LCA-modified mesoporous silica. The activation solution of S3 was added to 50 mL of L2 ethanol solution (10 mg / mL). The reaction was carried out at room temperature and 500 rpm in the dark for 24 h. The mixture was centrifuged and washed to obtain 20% LCA-modified R500 mesoporous silica. R500-20% PEG 1000 and R500-20% PEG 1000 -LCA particle size and potential are as follows Figure 2 As shown, the surface potential gradually approaches electroneutrality from electronegativity, and then becomes electronegative again after DCA grafting. This may be because the carboxyl groups of LCA bind to the amino groups on the PEG surface, weakening the charge shielding effect of PEG itself. Characterization of the potential demonstrates the successful modification with LCA.

[0607] Example 106, S500-10% PEG 1000 - Synthesis of pamoic acid

[0608] S1: Preparation of carboxylic acid stock solution. Dissolve 100 mg of pamoic acid in 10 mL of DMF.

[0609] S2: Carboxyl activation. NHS and EDC were added sequentially to the S1 solution, and the solution was activated at room temperature for 15 min.

[0610] S3: Pamoic acid-modified mesoporous silica. The activation solution prepared in S2 was added to the 10% PEG prepared in Example 5. 1000 The modified S500 was reacted in an ethanol solution at room temperature for 2 hours. The mixture was then centrifuged and washed to obtain pamoic acid-modified S500-10% PEG.1000 .

[0611] Example 107, S500-20% PEG 1000 - Synthesis of pamoic acid

[0612] S1: Preparation of carboxylic acid stock solution. Dissolve 100 mg of pamoic acid in 10 mL of DMF.

[0613] S2: Carboxyl activation. NHS and EDC were added sequentially to the S1 solution, and the solution was activated at room temperature for 15 min.

[0614] S3: Pamoic acid-modified mesoporous silica. The activating solution prepared in S2 was added to the 20% PEG prepared in Example 15. 1000 The modified S500 was reacted in an ethanol solution at room temperature for 2 hours. The mixture was then centrifuged and washed to obtain pamoic acid-modified S500-20% PEG. 1000 .

[0615] Example 108, S500-30% PEG 1000 - Synthesis of pamoic acid

[0616] S1: Preparation of carboxylic acid stock solution. Dissolve 100 mg of pamoic acid in 10 mL of DMF.

[0617] S2: Carboxyl activation. NHS and EDC were added sequentially to the S1 solution, and the solution was activated at room temperature for 15 min.

[0618] S3: Pamoic acid-modified mesoporous silica. The activating solution prepared in S2 was added to the 30% PEG prepared in Example 25. 1000 The modified S500 was reacted in an ethanol solution at room temperature for 2 hours. The mixture was then centrifuged and washed to obtain pamoic acid-modified S500-30% PEG. 1000 .

[0619] Example 109, S1000-10% PEG 1000 - Synthesis of pamoic acid

[0620] S1: Preparation of carboxylic acid stock solution. Dissolve 100 mg of pamoic acid in 10 mL of DMF.

[0621] S2: Carboxyl activation. NHS and EDC were added sequentially to the S1 solution, and the solution was activated at room temperature for 15 min.

[0622] S3: Pamoic acid-modified mesoporous silica. The activation solution prepared in S2 was added to the 10% PEG prepared in Example 35. 1000 The modified S1000 was reacted in an ethanol solution at room temperature for 2 hours. The mixture was then centrifuged and washed to obtain peramic acid-modified S1000-10% PEG.1000 .

[0623] Example 110, S1000-20% PEG 1000 - Synthesis of pamoic acid

[0624] S1: Preparation of carboxylic acid stock solution. Dissolve 100 mg of pamoic acid in 10 mL of DMF.

[0625] S2: Carboxyl activation. NHS and EDC were added sequentially to the S1 solution, and the solution was activated at room temperature for 15 min.

[0626] S3: Pamoic acid-modified mesoporous silica. The activating solution prepared in S2 was added to the 20% PEG prepared in Example 45. 1000 The modified S1000 was reacted in an ethanol solution at room temperature for 2 hours. The mixture was then centrifuged and washed to obtain pamoic acid-modified S1000-10% PEG. 1000 .

[0627] Example 111, S1000-30% PEG 1000 - Synthesis of pamoic acid

[0628] S1: Preparation of carboxylic acid stock solution. Dissolve 100 mg of pamoic acid in 10 mL of DMF.

[0629] S2: Carboxyl activation. NHS and EDC were added sequentially to the S1 solution, and the solution was activated at room temperature for 15 min.

[0630] S3: Pamoic acid-modified mesoporous silica. The activating solution prepared in S2 was added to the 30% PEG prepared in Example 55. 1000 The modified S1000 was reacted in an ethanol solution at room temperature for 2 hours. The mixture was then centrifuged and washed to obtain pamoic acid-modified S1000-10% PEG. 1000 .

[0631] Example 112, R500-10% PEG 1000 - Synthesis of pamoic acid

[0632] S1: Preparation of carboxylic acid stock solution. Dissolve 100 mg of pamoic acid in 10 mL of DMF.

[0633] S2: Carboxyl activation. NHS and EDC were added sequentially to the S1 solution, and the solution was activated at room temperature for 15 min.

[0634] S3: Pamoic acid-modified mesoporous silica. The activating solution prepared in S2 was added to the 10% PEG prepared in Example 65. 1000 The modified R500 was reacted in an ethanol solution at room temperature for 2 hours. The mixture was then centrifuged and washed to obtain pamucic acid-modified R500-10% PEG.1000 .

[0635] Example 113, R500-20% PEG 1000 - Synthesis of pamoic acid

[0636] S1: Preparation of carboxylic acid stock solution. Dissolve 100 mg of pamoic acid in 10 mL of DMF.

[0637] S2: Carboxyl activation. NHS and EDC were added sequentially to the S1 solution, and the solution was activated at room temperature for 15 min.

[0638] S3: Pamoic acid-modified mesoporous silica. The activation solution prepared in S2 was added to the 20% PEG prepared in Example 70. 1000 The modified R500 was reacted in an ethanol solution at room temperature for 2 hours. The mixture was then centrifuged and washed to obtain pamolic acid-modified R500-20% PEG. 1000 .

[0639] Example 114, R500-30% PEG 1000 - Synthesis of pamoic acid

[0640] S1: Preparation of carboxylic acid stock solution. Dissolve 100 mg of pamoic acid in 10 mL of DMF.

[0641] S2: Carboxyl activation. NHS and EDC were added sequentially to the S1 solution, and the solution was activated at room temperature for 15 min.

[0642] S3: Pamoic acid-modified mesoporous silica. The activating solution prepared in S2 was added to the 30% PEG prepared in Example 75. 1000 The modified R500 was reacted in an ethanol solution at room temperature for 2 hours. The mixture was then centrifuged and washed to obtain pamoic acid-modified R500-30% PEG. 1000 .

[0643] Example 115: Gastrointestinal stability of representative carrier drug conjugates

[0644] Take 3.84 mL of concentrated hydrochloric acid, add 800 mL of distilled water, add 10 g of pepsin, sonicate to mix thoroughly, dilute with water to 1 L, and filter through a 0.22 μm aqueous filter to prepare simulated gastric juice. Take 0.68 g of potassium dihydrogen phosphate, add 50 mL of water to dissolve it, and adjust the pH to 6.8 with 0.1 mol / L sodium hydroxide solution. Take 500 μL of the above solution, add 10 mg of trypsin, mix well, dilute to 1 mL, and finally filter through a 0.22 μm aqueous filter to obtain simulated intestinal juice.

[0645] The following carrier-drug conjugates were used in experiments: S500-10% PEG1000-DCA from Example 5, S500-20% PEG1000-DCA from Example 15, S500-30% PEG1000-DCA from Example 25, R500-10% PEG1000-DCA from Example 65, R500-20% PEG1000-DCA from Example 70, and R500-30% PEG1000-DCA from Example 75. DCA, S1000-10% PEG1000-DCA of Example 35, S1000-20% PEG1000-DCA of Example 45, S1000-30% PEG1000-DCA of Example 55, R1000-10% PEG1000-DCA of Example 95, R1000-20% PEG1000-DCA of Example 96, R1000-30% PEG1000-DCA of Example 97, etc. Example 10: S500-10% PEG2000-DCA; Example 20: S500-20% PEG2000-DCA; Example 30: S500-30% PEG2000-DCA; Example 80: R500-10% PEG2000-DCA; Example 85: R500-20% PEG2000-DCA; Example 90: R500-30% PEG2000-DCA; Example 40: S1000 -10% PEG2000-DCA, Example 50: S1000-20% PEG2000-DCA, Example 60: S1000-30% PEG2000-DCA, Example 98: R1000-10% PEG2000-DCA, Example 99: R1000-20% PEG2000-DCA, Example 100: R1000-30% PEG2000-DCA, Example 105: R500-20% PEG 1000 -LCA. The above-mentioned carrier drug conjugates were resuspended in simulated gastrointestinal fluid and incubated in a shaker at 37°C for 6 hours. The carrier solution was then diluted with purified water to 0.5 mg / mL. After being gently pipetted to ensure uniform dispersion, changes in particle size, potential, and polydispersity index (PDI) were monitored using a particle size analyzer.

[0646] The results are as follows Figure 5 As shown, the particle size and potential of these carrier-drug conjugates did not change significantly within 6 hours. This example demonstrates that the carrier-drug conjugates of the present invention possess gastrointestinal stability.

[0647] Example 116: Verification of the Cellular Non-Absorption of Representative Carrier-Drug Conjugates

[0648] The following carrier-drug conjugates were used in experiments: S500-10% PEG1000-DCA from Example 5, S500-20% PEG1000-DCA from Example 15, S500-30% PEG1000-DCA from Example 25, R500-10% PEG1000-DCA from Example 65, R500-20% PEG1000-DCA from Example 70, and R500-30% PEG1000-DCA from Example 75. DCA, S1000-10% PEG1000-DCA of Example 35, S1000-20% PEG1000-DCA of Example 45, S1000-30% PEG1000-DCA of Example 55, R1000-10% PEG1000-DCA of Example 95, R1000-20% PEG1000-DCA of Example 96, R1000-30% PEG1000-DCA of Example 97, etc. Example 10: S500-10% PEG2000-DCA; Example 20: S500-20% PEG2000-DCA; Example 30: S500-30% PEG2000-DCA; Example 80: R500-10% PEG2000-DCA; Example 85: R500-20% PEG2000-DCA; Example 90: R500-30% PEG2000-DCA; Example 40: S1000 -10% PEG2000-DCA, Example 50: S1000-20% PEG2000-DCA, Example 60: S1000-30% PEG2000-DCA, Example 98: R1000-10% PEG2000-DCA, Example 99: R1000-20% PEG2000-DCA, Example 100: R1000-30% PEG2000-DCA, Example 105: R500-20% PEG 1000 -LCA. In addition, a control group, S100-20% PEG1000-DCA, is required. Its preparation method is as follows: Weigh 110 mg of triethanolamine (TEA) and 0.4 g of N,N,N-trimethyl-1-hexadecylammonium bromide (CTAB), add 20 mL of deionized water, sonicate to dissolve, and incubate in an oil bath for 1 hour (95℃, above 1000 rpm). Add 1.5 mL of tetraethyl orthosilicate (TEOS), continue the reaction for 1 hour, and then centrifuge (20 minutes, 13000 rpm). Wash the precipitate three times with anhydrous ethanol, and finally resuspend it in 30 mL of ethanol. Add 1 mL of concentrated hydrochloric acid and incubate in an oil bath overnight (60℃, above 1000 rpm). Wash the precipitate sequentially with anhydrous ethanol and deionized water to obtain S100. Disperse 500 mg of S100 in a mixture of 340 mL of water and 200 mL of ethanol, and adjust the pH to 4.0 with dilute hydrochloric acid. Add 20% silane-PEG1000 The NH2 ethanol solution was added, and then ethanol solution was added to make the ethanol:water volume ratio of the reaction system 1:2. The reaction was carried out at 65℃ and 800rpm for 24h. The carrier was washed by centrifugation and resuspended in anhydrous ethanol to obtain 20% PEG. 1000 -NH2-modified S100 solution. Dissolve 109.8 mg DCA in 11.0 mL DMF. Then add 21 mg NHS and 56 mg EDC sequentially, activate at room temperature for 15 min, and adjust pH to 5.8. Add this activated solution to 50 mL of 20% PEG. 1000 The mixture was reacted with an NH2-modified S100 solution (10 mg / mL) at 500 rpm in the dark for 24 h at room temperature. The mixture was then centrifuged and washed to obtain S100-20% PEG1000-DCA.

[0649] Caco-2 cells were seeded in 24-well plates and cultured until adherent growth was achieved. 200 μL of blank culture medium was added, and the cells were incubated at 37°C for 20 min. The blank culture medium was then removed, and different carrier drug conjugates (300 μL, 50 μg / mL) were added to each well, followed by incubation for 2 h. After discarding the supernatant, the cells were washed with PBS, and RIPA lysis buffer was added, followed by lysis on a shaker at 37°C for 30 min. The cell lysates were collected and centrifuged at 4,000 rpm for 10 min. 200 μL of the supernatant was used to measure fluorescence in a 96-well black plate. Separately, 20 μL of the supernatant was added to 200 μL of BCA working solution, and the cells were incubated on a shaker at 37°C for 30 min. The absorbance of each well was measured at 562 nm using a microplate reader.

[0650] E12 cells were divided into 2×10 5 Cells were seeded at a density of cells / well in 24-well Transwell chambers and cultured for 18 days for in vitro differentiation. Cell monolayers were first equilibrated in HBSS for 20 min, followed by the addition of 0.5 mL of 50 μg / mL carrier-drug conjugate to the top side. At 8 h, 200 μL of sample was removed from the substrate, and fluorescence intensity was measured using a microplate reader.

[0651] The results are as follows Figure 6 As shown, compared to S100-20% PEG1000-DCA, the cellular uptake of various carrier drug conjugates is very low, and the transcellular transport in the E12 cell monolayer is very low, with the average fluorescence intensity detected being close to 0. This example demonstrates that the carrier drug conjugates of the present invention are difficult to absorb or transport through the intestinal monolayer cells, exhibiting non-absorbability.

[0652] Example 117: TGR5 protein levels adhering to the surface of representative carrier drug conjugates

[0653] First, a cell lysis buffer containing TGR5 protein was prepared. STC-1 cells were seeded in culture dishes and cultured at 37°C and 5% CO2 for 24 h. When the cells reached 80-90% confluence, they were washed with PBS to remove debris and unattached cells. 1 mL of RIPA cell lysis buffer containing 1% protease and phosphatase inhibitors was added to the culture dish, and the cells were lysed on ice for 30 min. The lysis buffer was collected and centrifuged at 14,500 rpm for 10 min at 4°C. The supernatant was collected, and 20 μL of the supernatant was used to determine the protein concentration using the BCA method. The remaining supernatant was used for subsequent protein carrier binding experiments.

[0654] The following drug conjugates were used to characterize the TGR5 protein-particle binding ability: S500-10% PEG1000-DCA (Example 5), S500-20% PEG1000-DCA (Example 15), S500-30% PEG1000-DCA (Example 25), R500-10% PEG1000-DCA (Example 65), R500-20% PEG1000-DCA (Example 70), and R500- 30% PEG1000-DCA, S1000-10% PEG1000-DCA of Example 35, S1000-20% PEG1000-DCA of Example 45, S1000-30% PEG1000-DCA of Example 55, R1000-10% PEG1000-DCA of Example 95, R1000-20% PEG1000-DCA of Example 96, R1000-30% PEG1000 of Example 97 0-DCA, S500-10% PEG2000-DCA of Example 10, S500-20% PEG2000-DCA of Example 20, S500-30% PEG2000-DCA of Example 30, R500-10% PEG2000-DCA of Example 80, R500-20% PEG2000-DCA of Example 85, R500-30% PEG2000-DCA of Example 90, S10 of Example 40 Example 50: S1000-10% PEG2000-DCA; Example 60: S1000-30% PEG2000-DCA; Example 98: R1000-10% PEG2000-DCA; Example 99: R1000-20% PEG2000-DCA; Example 100: R1000-30% PEG2000-DCA; Example 105: R500-20% PEG2000-DCA. 1000-LCA. Mix 0.1 mL of the carrier drug conjugate with 1 mL of the above-mentioned protein solution containing 1 mg / mL TGR5 and incubate at 37 °C for 2 h. Place the incubated mixture in 1 mL of 0.7 M sucrose solution and centrifuge at 4 °C and 20,000 g for 1 h to separate the carrier drug conjugate-protein complex precipitate from the mixture. Resuspend the precipitate in 50 μL of lysis buffer (containing 4% Triton × 100 and 2% SDS) and sonicate in an ice bath for 1 h to release the particle-bound protein. Then centrifuge the lysis buffer at 4 °C and 20,000 g for 0.5 h. Collect the supernatant, use 20 μL of the supernatant for BCA, and use the remaining supernatant for subsequent protein immunoblotting. First, boil the supernatant sample in boiling water for 10 min, and then separate the protein by 15% SDS polyacrylamide gel electrophoresis. Proteins were transferred to polyvinylidene fluoride (PVDF) membranes and blocked with blocking buffer. The membranes were then incubated overnight at 4°C with rabbit TGR5 or rabbit GAPDH primary antibody. The membranes were washed five times with Tris-buffered saline containing 1% Tween 20. Afterward, the membranes were stained with goat anti-rabbit IgG-HRP secondary antibody for 1 hour at room temperature, and then washed five times with Tris-buffered saline containing 1% Tween 20. The membranes were then removed, chemiluminescent buffer was added, and the membranes were imaged using a gel imaging system. Images were analyzed using ChemiScope Analysis software and GraphPad Prism 9.0 software.

[0655] The results are as follows Figure 7 As shown in the example, this embodiment demonstrates that the carrier drug conjugate of the present invention has good TGR5 binding ability.

[0656] Example 118: MST determination of the dissociation constant of representative carrier drug conjugates with TGR5 receptor protein.

[0657] The following representative carrier-drug conjugates were used in the experiments: S500-10% PEG1000-DCA from Example 5, S500-20% PEG1000-DCA from Example 15, S500-30% PEG1000-DCA from Example 25, R500-10% PEG1000-DCA from Example 65, R500-20% PEG1000-DCA from Example 70, and R500-30% PEG1000 from Example 75. 0-DCA, S1000-10% PEG1000-DCA of Example 35, S1000-20% PEG1000-DCA of Example 45, S1000-30% PEG1000-DCA of Example 55, R1000-10% PEG1000-DCA of Example 95, R1000-20% PEG1000-DCA of Example 96, and R1000-30% PEG1000-DCA of Example 97 Example 10: S500-10% PEG2000-DCA; Example 20: S500-20% PEG2000-DCA; Example 30: S500-30% PEG2000-DCA; Example 80: R500-10% PEG2000-DCA; Example 85: R500-20% PEG2000-DCA; Example 90: R500-30% PEG2000-DCA; Example 40: S100 0-10% PEG2000-DCA, S1000-20% PEG2000-DCA of Example 50, S1000-30% PEG2000-DCA of Example 60, R1000-10% PEG2000-DCA of Example 98, R1000-20% PEG2000-DCA of Example 99, R1000-30% PEG2000-DCA of Example 100, R500-20% PEG of Example 105 1000-LCA. First, 293T cells were seeded in 6-well plates. After 48 hours, the culture medium was replaced with antibiotic-free medium, and transfection began 24 hours later. 3.6 μL of TGR5 plasmid was mixed with 300 μL of blank medium, and 6 μL of liposome nucleic acid transfection reagent was mixed with 300 μL of blank medium. After standing for 4 min, this mixture was added to the plasmid solution and stood for 20 min before being added to each well of the 6-well plate. This was the volume per well. 18 h after transfection, the supernatant was discarded, and 300 μL of RIPA lysis buffer (containing 1% PMSF) was added to each well. Lysis was performed on ice for approximately 5 min. The cell lysates were collected and centrifuged for 15 min (14500 rpm, 4℃), retaining the supernatant. Protein expression levels were verified by detecting total fluorescence through a 488 nm excitation filter and a 509 nm emission filter. The protein supernatant was diluted 10-fold with PBS to provide optimal fluorescence levels. Each vector drug conjugate was stepwise diluted into 16 gradients using PBS. After incubating 10 μL of cell lysis buffer with 10 μL of different concentrations of carrier drug conjugates for 20 min, all samples were loaded into standard glass capillaries of the Temper mo-ko22 nanotube. Microscale thermoelectrophoresis was performed by scanning each capillary to analyze the binding affinity of different carrier drug conjugates to the TGR5 receptor protein. The binding affinity test method for R500-20% PEG1000-pamoate to GPR35 protein in Example 113 was similar to that described above.

[0658] The results are as follows Figure 8 As shown, the dissociation constants of the carrier-drug conjugates are all smaller than those of small molecules. This example demonstrates that the carrier-drug conjugates of the present invention have good TGR5 binding ability.

[0659] Example 119: Representative drug conjugate efficacy at the cellular level (GLP-1 level)

[0660] The following carrier-drug conjugates were used in experiments: S500-10% PEG1000-DCA from Example 5, S500-20% PEG1000-DCA from Example 15, S500-30% PEG1000-DCA from Example 25, R500-10% PEG1000-DCA from Example 65, R500-20% PEG1000-DCA from Example 70, and R500-30% PEG1000-DCA from Example 75. DCA, S1000-10% PEG1000-DCA of Example 35, S1000-20% PEG1000-DCA of Example 45, S1000-30% PEG1000-DCA of Example 55, R1000-10% PEG1000-DCA of Example 95, R1000-20% PEG1000-DCA of Example 96, R1000-30% PEG1000-DCA of Example 97, etc. Example 10: S500-10% PEG2000-DCA; Example 20: S500-20% PEG2000-DCA; Example 30: S500-30% PEG2000-DCA; Example 80: R500-10% PEG2000-DCA; Example 85: R500-20% PEG2000-DCA; Example 90: R500-30% PEG2000-DCA; Example 40: S1000 -10% PEG2000-DCA, Example 50: S1000-20% PEG2000-DCA, Example 60: S1000-30% PEG2000-DCA, Example 98: R1000-10% PEG2000-DCA, Example 99: R1000-20% PEG2000-DCA, Example 100: R1000-30% PEG2000-DCA, Example 105: R500-20% PEG 1000 -LCA.

[0661] STC-1 cells were divided into 8×10 5 Cells were seeded at a density of cells / well in 12-well plates and cultured for 48 h. Before drug administration, the culture medium was discarded, and the cells were washed with KR buffer. After incubating the cells with each drug conjugate for 120 min, the cell supernatant was collected, centrifuged at 2,000 g for 10 min, and the supernatant was collected. The GLP-1 secretion level was detected using a mouse GLP-1 ELISA kit.

[0662] The results are as follows Figure 9As shown, the rod-shaped formulations were superior to the spherical formulations; the R1000 formulations were slightly superior to the R500 formulations; and the 20% PEG-modified and 30% PEG-modified formulations were superior to the 10% PEG-modified formulations. This example demonstrates that the carrier-drug conjugate of the present invention has a good ability to promote GLP-1 secretion.

[0663] R500-20% PEG in Examples 120 and 70 1000 - DCA transport in the digestive tract of db / db mice

[0664] db / db mice were fasted for 18 hours beforehand, with free access to water. They were then orally administered RITC-labeled R500-20% PEG. 1000 -DCA, administered at a dose of 10 mg. Mice were euthanized at 1, 2, 3, 6, 9, and 12 hours post-administration. The digestive tract and major organs of the mice were removed and placed on a blackboard. In vivo imaging was used to image the digestive tract and major organs (including heart, liver, spleen, lungs, and kidneys). The R500-20% PEG ratio was statistically analyzed using the in vivo imaging analysis module and GraphPad Prism 9.0. 1000 - Distribution of DCA in the digestive tract and its retention in intestinal segments.

[0665] Among them, RITC-labeled R500-20% PEG 1000 -DCA was prepared as follows: 10 mg of red fluorescent powder RITC was dissolved in 2 mL of anhydrous ethanol, followed by the addition of 2 μL of 3-aminopropyltriethoxysilane (APTES) under stirring. The reaction was carried out at room temperature and 200 rpm in the dark for 8 h. The reaction was then carried out in 5 mL of 2 mg / mL R500-20% PEG. 1000 Add APTES-RITC reaction solution to an ethanol solution of DCA or R500 and react at 40°C and 200 rpm for 24 h. Then centrifuge at 12000 rpm for 15 min to obtain the precipitate, and wash three times with anhydrous ethanol to obtain the final product.

[0666] The results are as follows Figure 10 As shown, R500-20% PEG 1000 DCA has non-absorbable properties in the digestive tract, rapidly reaching the ileum within 2 hours and remaining in the ileum and colon for an extended period of approximately 9 hours. This example demonstrates R500-20% PEG. 1000 -DCA has the characteristics of non-absorbability and long-lasting intestinal retention.

[0667] R500-20% PEG from Examples 121 and 70 1000 -DCA promotes TGR5 accumulation in db / db mice

[0668] db / db mice were fasted for 18 hours prior to the procedure. Subsequently, the mice were anesthetized and kept warm on a temperature-controlled mat. The abdomen was then dissected, the colon exposed, and a 1 cm segment of the colon was ligated with surgical sutures. A local injection of 100 μL of 200 μg / mL R500-20% PEG was then administered. 1000 - A PBS suspension of DCA was applied, followed by suturing of the abdominal cavity. Two hours later, the ligated colonic segment was removed, and approximately 1 mm was excised. 3 Tissue samples were slowly washed with cold PBS and fixed with a fixative for immunoelectron microscopy. Samples were resin-embedded, ultrasliced, and loaded onto 150-mesh nickel grids. The grids were then washed three times with 30 μL PBS droplets and placed face-up in blocking buffer containing 0.1% Tween 20 and 1% skim milk in PBS and incubated at room temperature for 1 h. Next, the nickel grids were incubated overnight at 4°C with 25 μL of TGR5 antibody (1:500 dilution). The nickel grids were then washed three times with 25 μL of PBS buffer containing 0.1% Tween 20 each time. The grids were incubated with 25 μL of gold-labeled goat anti-rabbit secondary antibody (1:30 dilution) at room temperature for 1 h. The nickel grids were washed three times with PBS buffer and double-distilled water, respectively. Excess liquid was blotted from the grid edges with filter paper, then negatively stained and imaged under a transmission electron microscope. Finally, the images were analyzed using Fiji software.

[0669] The results are as follows Figure 11 As shown, compared with the control group, R500-20% PEG 1000 - DCA treatment significantly increased the aggregation of gold-labeled TGR5 in colonic tissue, indicating that R500-20% PEG 1000 -DCA still retains its ability to induce TGR5 aggregation in intestinal cells in vivo. This example demonstrates the effectiveness of the R500-20% PEG described in Example 70. 1000 -DCA has the ability to promote TGR5 aggregation.

[0670] R500-20% PEG in Examples 122 and 70 1000 DCA promotes the secretion of GLP-1 in db / db mice.

[0671] db / db mice were fasted overnight. Based on their fasting blood glucose levels and body weight, the mice were divided into four groups (n=5). Subsequently, these mice received subcutaneous injections of liraglutide or gavage with a blank solution, DCA, or R500-20% PEG. 1000-DCA. Blood samples were collected via orbital sampling at 2, 6, and 10 h post-administration and placed in centrifuge tubes containing 1 μg / mL DPP IV inhibitor and 0.1% heparin sodium. The samples were then immediately centrifuged at 3,000 rpm and 4°C for 10 min. Serum GLP-1 levels were measured using a mouse GLP-1 ELISA kit.

[0672] The results are as follows Figure 12 As shown, in the R500-20% PEG1000-DCA group, the blood GLP-1 level increased significantly compared to the DCA group or the liraglutide group from 2 to 6 hours after administration, and this increase remained relatively significant even after 10 hours. The blood GLP-1 levels in the R500-20% PEG1000-DCA group at 2, 6, and 10 hours were 3.0, 3.6, and 3.7 times that of the control group, respectively. This example demonstrates that the R500-20% PEG1000-DCA described in Example 70 can provide important mechanistic support for its sustained hypoglycemic effect by continuously promoting GLP-1 secretion.

[0673] R500-20% PEG in Examples 123 and 70 1000 -DCA in OGTT in db / db mice

[0674] Prior to the oral glucose tolerance test (OGTT), db / db mice and healthy C57 mice were fasted overnight, with free access to water. Based on their fasting blood glucose levels and body weight, db / db mice were divided into four groups (n=5): (1) blank solution (Control, 400 μL, ig); (2) DCA (50 mg / kg, ig); (3) liraglutide (LIR, 123 μg / kg, sc); and (4) R500-20% PEG. 1000 -DCA (963 mg / kg, ig). C57 healthy mice were administered a blank solution via gavage. Two hours prior to glucose administration, mice received either a subcutaneous injection of liraglutide or oral administration of different formulations. Subsequently, mice were administered 2 g / kg glucose via gavage. Blood samples were collected from the tail at 0, 15, 30, 60, 90, and 120 min after glucose administration, and blood glucose levels were measured using a glucometer. The blood glucose concentration-time curve and area under the curve (AUC) from 0 to 120 min were analyzed using GraphPad Prism 9.0. 0-120min ).

[0675] The results are as follows Figure 13 As shown, R500-20% PEG 1000- Blood glucose levels in the DCA group decreased rapidly after treatment and returned to normal levels approximately 1 hour after glucose gavage. DCA, liraglutide, and R500-20% PEG 1000 AUC of blood glucose in mice in the DCA group 0-120min The levels were reduced to 70.9%, 70.1%, and 63.7% in the Control group, respectively. This example illustrates that R500-20% PEG 1000 A single oral dose of DCA significantly reduced blood glucose and improved oral glucose tolerance in db / db mice, demonstrating a stronger and more sustained hypoglycemic effect than liraglutide or DCA.

[0676] R500-20% PEG in Examples 124 and 70 1000 -Long-term efficacy of DCA in db / db mice

[0677] Twenty db / db mice were randomly selected and divided into four groups, and treated for 30 days to evaluate R500-20% PEG. 1000 -Long-term therapeutic effects of DCA. Mice were administered subcutaneously with semaglutide (Sema, 102.5 μg / kg / week, equivalent to 0.5 mg / week in humans) or by gavage with the following formulations: control (blank solution), free DCA solution (50 mg / kg / day), or R500-20% PEG. 1000 -DCA solution (963 mg / kg / d, equivalent to 50 mg / kg / d DCA). Five healthy C57 mice were administered the blank solution via gavage. The day before administration was designated as Day 0, and the first administration was designated as Day 1, continuing for 30 days. During days 0-30, blood was collected daily at 3 PM via the tip of a lancet, and blood glucose levels were recorded once using a glucometer. To monitor continuous changes in blood glucose, all mice were observed for 5 days after discontinuation of the drug, and an oral glucose tolerance test (OGTT) was performed on day 35. Mice were fasted overnight beforehand, then orally administered 2 g / kg glucose, and blood glucose levels were measured at 15, 30, 60, and 120 minutes after glucose gavage.

[0678] The results are as follows Figure 14 As shown, R500-20% PEG 1000 Compared to the other three groups, the DCA group gradually reduced blood glucose levels in mice, reaching 11.1 mmol / L after 17 days of administration, demonstrating a more sustained hypoglycemic effect. OGTT results after the long-term administration period showed that R500-20% PEG... 1000 Mice in the DCA group exhibited a rapid decrease in blood glucose levels after glucose administration, returning to normal levels after approximately 2 hours. Compared to the control group, R500-20% PEG...1000 -DCA group's AUC 0-120min The glucose tolerance was reduced by 38.4%. Mice in other treatment groups showed similar glucose tolerance to the control group, with no significant improvement. These results highlight the effectiveness of R500-20% PEG. 1000 - DCA continues to improve type 2 diabetes even after long-term treatment, highlighting its potential advantages in the long-term management of type 2 diabetes.

[0679] This example demonstrates the effectiveness of the R500-20% PEG described in Example 70. 1000 DCA has significant and sustained therapeutic potential in long-term glycemic control of type 2 diabetes.

[0680] R500-20% PEG from Examples 125 and 70 1000 - DCA transit in the digestive tract of Bama pigs

[0681] To specifically track R500-20% PEG in live pigs 1000 -DCA, referring to the methods reported in the literature for selecting gadolinium element-labeled carriers (Hsiao, JK et al., Small, 2008; Bouchoucha, M. et al., Advanced Functional Materials, 2014; Laprise-Pelletier, M. et al., Journal of Materials Chemistry B, 2015; Ren, HH et al., Journal of Materials Chemistry B, 2017), prepared and characterized gadolinium-labeled non-absorbable carrier drug conjugates. First, 2g of R500-20% PEG was used. 1000 DCA was dispersed in 400 mL of ethanol, and 400 μL of APTES was added. The mixture was reacted at 800 rpm and 40 °C for 8 h. R500-20% PEG was obtained by centrifugation and washing. 1000 -DCA-APTES, then 1g DTPA was added to 340mL dimethyl sulfoxide and stirred for 2h to obtain R500-20% PEG. 1000 -DCA-DTPA. Next, add R500-20% PEG. 1000 DCA-DTPA was dispersed in 75% methanol containing 7.4 g GdCl3·6H2O and stirred at 40°C for 18 h. Gd-labeled R500-20% PEG was obtained by centrifugation and washing. 1000 -DCA (named Gd-R500-20% PEG) 1000-DCA). Finally, 3 mL suspensions were prepared at concentrations of 0.25 mg / mL, 0.5 mg / mL, 1 mg / mL, 5 mg / mL, and 10 mg / mL, added to 10 mL centrifuge tubes, and analyzed on a 3.0 T magnetic resonance high-field scanner. T1-weighted images were acquired using a fast phase-scrambled gradient echo sequence with parameters including TR = 7 ms, TE = 3.37 ms, flip angle of 15 degrees, flip time of 100 ms, field of view of 400 × 400 mm, matrix of 205 × 256, and pixel size of 0.98 × 0.78 × 1 mm.

[0682] To evaluate R500-20% PEG 1000 -Device endothelial (DCA) study investigated the digestive tract transport characteristics in a large animal model, using Bama pigs weighing approximately 20 kg. To minimize the impact of food and feces on imaging, the pigs were fasted for 24 hours, followed by gavage administration of 1,400 mL of PEG electrolyte solution, administered in eight divided doses. During the final gavage, 15 mL of dimethyl silicone oil was added to the PEG electrolyte solution to promote defecation. The pigs were fasted throughout the experiment but were allowed to drink water containing 5% glucose. To obtain initial images of the digestive tract, the pigs were subcutaneously injected with 3 mg / kg of styraxate, and imaging was performed after anesthesia. Two hours after anesthesia, when the pigs awoke, they were administered Gd-R500-20% PEG via gavage. 1000 -DCA (100 mg / kg). Pigs were pre-trained for MRI noise and imaging and remained awake during the experiment. Subsequently, MRI imaging was performed on pigs in a prone position at 1, 2, 3, 6, 9, 12, and 24 hours after administration. T1-weighted images of coronal sections were acquired using a fast breaking gradient echo sequence with the following imaging parameters: TR = 2.6 ms, TE = 1.11 ms, flip angle of 10 degrees, inversion time of 110 ms, and field of view of 360 × 320 mm. T2-weighted images of coronal sections were acquired using a fast breaking gradient echo sequence with the following imaging parameters: TR = 14,000 ms, TE = 97.2 ms, and field of view of 380 × 300 mm.

[0683] The results are as follows Figure 15 As shown, R500-20% PEG 1000 DCA reaches the colon approximately 6 hours after administration and remains in the gastrointestinal tract for at least 24 hours. This is similar to results observed in mice, indicating that R500-20% PEG 1000 -DCA also exhibits good ileal and colonic retention characteristics in large animal models.

[0684] This example demonstrates the R500-20% PEG described in Example 70. 1000DCA exhibits excellent retention performance in the ileum and colon, and its precise tracking and localization are achieved through MRI imaging, demonstrating its applicability in large animal models and its potential for clinical translation.

[0685] R500-20% PEG in Examples 126 and 70 1000 -DCA in OGTT of Bama Pigs

[0686] To verify R500-20% PEG 1000 The hypoglycemic effect of DCA in a large animal model was demonstrated in an oral glucose tolerance test (OGTT) in Bama pigs. All pigs were fasted overnight before treatment. Then, the pigs were randomly divided into three groups (n=3), receiving either a subcutaneous injection of liraglutide (LIR, 14 μg / kg, equivalent to 123 μg / kg in mice), or oral administration of R500-20% PEG. 1000 - Blood glucose levels in pigs were monitored using a continuous glucose monitoring system (CGM) with DCA solution (110 mg / kg, equivalent to 963 μg / kg in mice) or a blank solution (Control). Three hours later, pigs were orally administered 4 g / kg of glucose solution. Blood glucose levels were continuously monitored for 3 hours using a CGM. The blood glucose concentration-time curve and area under the curve (AUC) from 0 to 180 minutes after glucose administration were analyzed using GraphPad Prism 9.0. 0–180min ).

[0687] The results are as follows Figure 16 As shown, 60 minutes after glucose administration, the blood glucose level in the blank solution group approximately doubled. In contrast, the level in the group receiving R500-20% PEG... 1000 In pigs with DCA, the rise in blood glucose levels after the glucose challenge was effectively mitigated, and the levels returned to baseline within 90 minutes, with an AUC of [missing value]. 0-180min Comparable to liraglutide. Therefore, the results indicate that R500-20% PEG... 1000 DCA demonstrated a significant ability to improve hyperglycemia in both small animal mouse models and large animal Bama pig models. This is for R500-20% PEG. 1000 - The consistent effect of DCA in different animal models provides experimental evidence, further supporting its potential for clinical application.

[0688] This example demonstrates the effectiveness of the R500-20% PEG described in Example 70. 1000 DCA has shown consistent and significant hypoglycemic effects in different animal models, supporting the feasibility of its clinical application.

[0689] R500-20% PEG in Examples 127 and 701000 Long-term efficacy of DCA in Bama pigs

[0690] Blood glucose levels were measured daily at 9:00 AM, followed by a subcutaneous injection of liraglutide (LIR, 14 μg / kg, equivalent to 123 μg / kg in mice) and oral administration of R500-20% PEG. 1000 -DCA (110 mg / kg, equivalent to 963 μg / kg for mice) or blank solution (Control). Three hours after administration, administer 4 g / kg glucose orally. Record daily food intake and body weight on days 0 and 14. Monitor blood glucose levels using a continuous glucose monitoring system (CGMS, FreeStyle Libre H, Abbott) at 0, 3, 3.25, 3.5, 3.75, 4, 4.5, 5, 5.5, 6, 6.5, and 7 hours after administration. On days 1, 7, and 14, collect blood samples from the posterior ear vein before feeding, 3 hours after administration. Serum samples were analyzed using a porcine GLP-1 ELISA kit (CUSABIO Technology, Wuhan, China) and a porcine insulin ELISA kit (Solarbio Science & Technology, Beijing, China).

[0691] like Figure 17 As shown, R500-20% PEG 1000 -DCA effectively alleviated hyperglycemia induced by diet and glucose stimulation in a 14-day repeated-dose study, with an AUC similar to that of the liraglutide group.

[0692] This example demonstrates the effectiveness of the R500-20% PEG described in Example 70. 1000 DCA has significant and sustained therapeutic potential in long-term glycemic control of type 2 diabetes.

[0693] R500-20% PEG from Examples 128 and 70 1000 DCA is non-absorbable in mice.

[0694] db / db mice were fasted for 18 hours prior to the event, during which they were allowed free access to water. Subsequently, they were orally administered RITC-labeled R500-20% PEG. 1000R500, prepared by DCA or Example 3, was administered at 10 mg / animal, with a total suspension volume of 0.4 mL. Major organs (heart, liver, spleen, lungs, and kidneys) were removed at 6 h for in vivo imaging, and the liver and kidneys were collected. Tissue samples were fixed with 4% paraformaldehyde for 4 h, then transferred to 30% sucrose solution and dehydrated overnight at 4°C. Next, the samples were sectioned using a cryostat to a thickness of 20 μm. Each section was stained with 10 μg / mL DAPI for 10 min and then washed three times with PBS. Finally, R500-20% PEG was observed under a confocal laser scanning microscope. 1000 - Distribution of DCA in major metabolic organs (liver and kidneys).

[0695] Among them, RITC-labeled R500-20% PEG 1000 DCA and R500 were prepared as follows: 10 mg of red fluorescent powder RITC was dissolved in 2 mL of anhydrous ethanol, followed by the addition of 2 μL of 3-aminopropyltriethoxysilane (APTES) under stirring. The reaction was carried out at room temperature and 200 rpm in the dark for 8 h. Then, 5 mL of 2 mg / mL R500-20% PEG was added. 1000 Add APTES-RITC reaction solution to an ethanol solution of DCA or R500 and react at 40°C and 200 rpm for 24 h. Then centrifuge at 12000 rpm for 15 min to obtain the precipitate, and wash three times with anhydrous ethanol to obtain the final product.

[0696] like Figure 18 As shown, R500-20% PEG 1000 -DCA did not enter the body, and no accumulation of carrier fluorescence was observed in tissue fluorescence staining of the liver and kidneys, thus proving that R500-20% PEG 1000 -DCA still exhibits non-absorbable properties in vivo.

[0697] R500-20% PEG in Examples 129 and 70 1000 Safety evaluation of DCA in mice

[0698] Twenty db / db mice and five C57 mice were randomly selected and divided into four groups, and treated for 30 days to evaluate R500-20% PEG. 1000 -Long-term safety of DCA. db / db mice were administered subcutaneously with semaglutide (Sema, 102.5 μg / kg / week, equivalent to 0.5 mg / week in humans) or by gavage with the following formulations: control (blank solution), free DCA solution (50 mg / kg / day), or R500-20% PEG.1000 DCA solution (963 mg / kg / d, equivalent to 50 mg / kg / d of DCA) was administered as a blank solution to C57 mice via gavage. After 30 days, blood samples were collected from the mice via orbital sampling and centrifuged at 3,000 rpm for 10 min at 4°C to obtain serum. Serum parameters related to liver function, such as alanine transaminase (ALT) and aspartate transaminase (AST), were analyzed using an automated blood analyzer. TNF-α and IL-6 were analyzed using an ELISA kit.

[0699] use Mice were anesthetized with 50 mg / mL (0.06 mL / 10 g), their abdomens were opened, and the gallbladders were photographed on striped paper using a smartphone camera. The length and width of the gallbladder were then measured using calipers, and the volume was calculated. Bile acids were collected from the gallbladder using a sterile insulin syringe. Mice were euthanized, and their livers were removed. Liver photographs were taken on striped paper using a smartphone camera, and the livers were immediately weighed. Liver samples were then preserved in paraformaldehyde for HE staining to observe liver morphology and Oil Red staining to observe liver lipid content. Liver tissue sections were photographed using a pathological section scanner, and liver lipid content was analyzed using Fiji software.

[0700] like Figure 19 As shown, R500-20% PEG 1000 -After long-term DCA administration, no gallbladder enlargement, bile accumulation, or liver toxicity was observed in db / db mice. R500-20% PEG 1000 - The DCA group also reduced the content of lipid droplets in the liver of mice. In addition, R500-20% PEG 1000 - In the DCA group, IL-6 and TNF-α levels were normal, indicating that R500-20% PEG 1000 DCA does not induce an inflammatory response in the body. This example illustrates that R500-20% PEG 1000 -DCA exhibits good biocompatibility in db / db mice and does not cause systemic toxicity.

Claims

1. A non-absorbable carrier drug conjugate comprising at least the structure shown in Figure 20: in, Component A is a non-absorbable carrier unit; component B is a linker; component C is a receptor binding unit. The non-absorbable carrier drug conjugate exhibits a hydration kinetic size change of no more than 5% within 24 hours in the physiological environment of the digestive tract, and the proportion absorbed within 24 hours in the physiological environment of the digestive tract is no more than 1%.

2. The non-absorbable carrier drug conjugate according to claim 1, wherein, The molar ratio of component B to component A is between 1,000 and 5,000,000, and the molar ratio of component C to component A is between 100 and 500,000; preferably, the molar ratio of component B to component A is between 5,000 and 4,000,000, and the molar ratio of component C to component A is between 300 and 400,000; more preferably, the molar ratio of component B to component A is between 10,000 and 4,000,000, and the molar ratio of component C to component A is between 500 and 400,000; most preferably, the molar ratio of component B to component A is between 20,000 and 3,000,000, and the molar ratio of component C to component A is between 2,000 and 300,000.

3. The non-absorbable carrier drug conjugate according to claim 1 or 2, wherein, The material of component A is selected from one or a combination of inorganic and organic materials; preferably, the material of component A is selected from one or a combination of silicon dioxide, graphene oxide, carbon quantum dots, carbon nanotubes, hydroxyapatite, titanium dioxide, metal-organic frameworks, covalent organic frameworks, and poly(lactic-co-glycolic acid); more preferably, the material of component A is selected from one or a combination of silicon dioxide, carbon nanotubes, and poly(lactic-co-glycolic acid); most preferably, the material of component A is silicon dioxide.

4. The non-absorbable carrier drug conjugate according to any one of claims 1-3, wherein: The hydration kinetics dimension of component A is between 400 nm and 10000 nm; preferably, the hydration kinetics dimension is between 400 nm and 2000 nm; and / or The aspect ratio of component A is ≥1.25; preferably, the aspect ratio is ≥1.5; more preferably, the aspect ratio is between 1.5 and 15.0; most preferably, the aspect ratio is between 6.0 and 12.

0.

5. The non-absorbable carrier drug conjugate according to any one of claims 1-4, wherein: The material of component B is selected from hydrophobic or hydrophilic polymers; preferably, the material of component B is selected from hydrophilic polymers; more preferably, the material of component B is selected from polyethylene glycol and its derivatives; most preferably, the material of component B is selected from polyethylene glycol derivatives, wherein one end of the polyethylene glycol molecule is modified to be amino; and / or The chain length of component B is in the range of 5–60 nm, which increases the hydration kinetic size of component A by 5–60 nm after component B is connected to component A; and / or The material of component B is polyethylene glycol and its derivatives containing more than 5 -CH2-CH2-O repeating units; preferably, the material of component B is polyethylene glycol and its derivatives containing 16 to 24 -CH2-CH2-O repeating units.

6. The non-absorbable carrier drug conjugate according to any one of claims 1-5, wherein: The material of component C is selected from G protein-coupled bile acid receptor (TGR5) agonists, G protein-coupled receptor 40 (GPR40) agonists, G protein-coupled receptor 120 (GPR120) agonists, G protein-coupled receptor 41 (GPR41) agonists, G protein-coupled receptor 43 (GPR43) agonists, G protein-coupled receptor 119 (GPR119) agonists, calcium ion-sensitive receptor (CaSR) agonists, and G protein-coupled receptor 35 (GPR35) agonists. One or a combination of agents; preferably, the material of component C is selected from TGR5 agonists and GPR35 agonists; more preferably, the material of component C is selected from bile acid compounds and their analogues, INT-777, zapustol, pamoic acid, lodusamide, YE120; even more preferably, the material of component C is selected from cholic acid, deoxycholic acid, chenodeoxycholic acid, ursodeoxycholic acid, lithocholic acid, pamoic acid, etc.; most preferably, the material of component C is selected from deoxycholic acid and pamoic acid; and / or The dissociation constant of the material of component C that undergoes a dissociation reaction with the corresponding receptor is no higher than 10 μM.

7. The non-absorbable carrier drug conjugate according to any one of claims 1-6, wherein, Components A and C are not directly connected. Components A and B are connected by covalent bonds or connecting functional groups. Components C and B are connected by covalent bonds or connecting functional groups. Preferably, the connecting functional group connecting components C and B is selected from one or a combination of amide bonds, ester bonds, thioether bonds, triazole bonds and carbamate bonds. Most preferably, the connecting functional group connecting components C and B is an amide bond.

8. The non-absorbable carrier drug conjugate according to any one of claims 1-7, wherein, The molar ratio of component B to component A is between 20,000 and 3,000,000, and the molar ratio of component C to component A is between 2,000 and 300,000. The material of component A is selected from silicon dioxide, and the size of component A is between 400 nm and 2,000 nm. The material of component B is polyethylene glycol and its derivatives, and the material of component C is selected from TGR5 agonist.

9. A pharmaceutical composition comprising: a carrier drug conjugate as described in any one of claims 1-8; and optionally pharmaceutically acceptable excipients. Preferably, the pharmaceutical composition is formulated as an oral preparation, such as tablets, capsules, granules, dispersible tablets, effervescent tablets, sustained-release preparations, controlled-release preparations, or enteric-coated preparations.

10. The use of the carrier drug conjugate as described in any one of claims 1-8 or the pharmaceutical composition as described in claim 9 in the preparation of a medicament for the prevention or treatment of a disease. Preferably, the disease is selected from diabetes, obesity, intestinal inflammation, and inflammatory bowel disease; more preferably, the disease is selected from diabetes.

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