Intelligent drug delivery system mediated by strong acting force and preparation method

Abstract

The invention discloses a strong acting force mediated intelligent drug delivery system and a preparation method. The system comprises a core-shell structure carrier matrix, a drug active component, a'main acting force + auxiliary acting force 'strong acting force unit and a surface functional layer, the preparation method comprises the following steps: preparing a mesoporous core layer and a polymer shell layer to form a core-shell carrier, mediating drug loading through a strong acting force unit, and finally modifying a targeting ligand and PEG (Polyethylene Glycol). The system triggers'pulse-slow release 'dual-mode release of a drug through double physiological microenvironment stimulation, a strong acting force unit improves the load stability (the leakage rate in 24 hours is smaller than or equal to 5%, and the encapsulation efficiency is larger than or equal to 75%), and targeting-anti-clearance modification enhances the targeting property (the target site enrichment amount is larger than or equal to 30%) and the circulation time (the half-life period is larger than or equal to 12 h). The preparation process is controllable, the quality control is clear, the requirements of GMP (good manufacturing practice) are met, the drug effect can be improved, the toxic and side effects can be reduced, and the traditional Chinese medicine composition is suitable for clinical transformation and industrialization.

Description

TECHNICAL FIELD

[0001] The present application belongs to the field of nanomedicine, and simultaneously intersects with the branch of novel drug delivery systems in the field of pharmaceutical preparation technology, and specifically relates to a strong force mediated intelligent drug delivery system and a preparation method, which is suitable for precise drug delivery for tumors, inflammatory diseases, metabolic diseases and the like. BACKGROUND

[0002] Current drug delivery technology faces multiple bottlenecks: traditional systems mostly rely on single hydrogen bond, physical adsorption and other weak force to load drugs, with low stability constant (usually <10 5 L / mol), leading to easy leakage of drugs during in vivo transport. For example, the drug leakage rate of liposome Doxil (reference: Nature Nanotechnology, 2020, 15(3): 206-217) based on physical adsorption is over 40% in 24 hours, with an encapsulation rate of only 50%-60%, which not only wastes drugs, but also increases the risk of non-target tissue toxicity. In addition, such systems have poor adaptability to different types of drugs such as small molecules, antibodies and nucleic acids, with an encapsulation rate of mostly 50%-70%, which is difficult to meet the clinical demand for efficient delivery.

[0003] In terms of drug release, existing systems are mostly single stimulus response (such as only pH response), which is easily disturbed by normal tissue microenvironment. For example, pH-sensitive polymer micelles (reference: Biomaterials, 2022, 280: 121302) release part of the drug at normal physiological pH (7.4), leading to an effective concentration at the target site maintained for ≤8h, which is difficult to precisely match the complex physiological conditions at the disease target site (such as acidity + high glutathione in tumors, acidity + high enzyme in inflammatory intestinal tract). At the same time, the release mode is mostly "burst release followed by failure" or "slow release with slow onset", which cannot maintain the drug concentration at the target site within the effective therapeutic window, with short treatment duration (usually ≤8h).

[0004] There are also deficiencies in targeting and bioavailability: most systems lack specific targeting design, or are only single-targeted, which are easily cleared by the mononuclear phagocyte system. For example, unmodified nanoparticles have an in vivo circulation half-life of ≤4h, and the enrichment amount at the target site is usually less than 15% (reference: Advanced Drug Delivery Reviews, 2019, 144: 103-118), with high distribution ratio in non-target organs, increasing the burden on liver and kidney and toxic side effects. At the same time, existing technologies are mostly designed for a single disease, with poor adaptability, and the preparation process parameters are ambiguous, and the quality control standards are not clear, which is difficult to meet the requirements of industrialization and clinical GMP, restricting clinical translation application.

[0005] The present application solves the above technical bottlenecks by introducing a strong force mediated intelligent drug delivery system, combining a core-shell carrier matrix, a dual stimulus response mechanism and surface functionalization modification, and provides a new strategy for efficient and safe drug delivery. SUMMARY

[0006] The technical problem to be solved by the present application is to overcome the defects of the above-mentioned technologies, and to provide a strong force mediated intelligent drug delivery system and a preparation method.

[0007] To solve the above technical problems, the technical solution provided by the present application is a strong force mediated intelligent drug delivery system and a preparation method: including a core-shell structure carrier matrix, a drug active ingredient, a strong force unit mediating the specific combination of the two, and a surface functionalization layer; the core layer of the core-shell structure carrier matrix is a mesoporous material (pore size 2-10 nm), the shell layer is a biocompatible polymer coating (thickness 10-50 nm), and the overall particle size is 80-400 nm and the dispersion coefficient PDI is ≤0.25; the strong force unit is composed of "main force + auxiliary force", the main force is selected from one of coordination bond and dynamic covalent bond, the auxiliary force is selected from at least one of hydrogen bond and π-π stacking interaction, and the main force can reversibly break / weaken in response to dual physiological microenvironment stimuli, and the auxiliary force cooperatively controls the drug release kinetics; under the dual physiological microenvironment stimuli, the main force breaks to make the drug-carrier binding force decrease by ≥60%, the auxiliary force maintains the stability of the carrier structure, and realizes the "pulse-slow release" dual-mode release of the drug active ingredient at the target site (24h cumulative release rate: 30%-50% released within 1h in the pulse phase, and 40%-60% released within 23h in the slow release phase).

[0008] As an improvement, the following steps are included:

[0009] (1) Core-shell carrier matrix preparation: ① Mesoporous core layer preparation: using tetraethyl orthosilicate / metal organic framework material as raw material, cetyltrimethylammonium bromide / polyetheramine as template agent, sol-gel reaction at pH 8.5-10.0 and temperature 45-60℃ for 8-12h, calcination (500-600℃, 4-6h) or solvent extraction to remove the template agent, to obtain the mesoporous core layer; ② Shell coating: dispersing the mesoporous core layer in a polymer solution (concentration 1-5mg / mL), under nitrogen protection and stirring speed 300-500rpm, adding crosslinking agent (mass ratio of polymer 1:5-1:10), reacting at 30-40℃ for 6-8h, centrifuging (8000-12000rpm, 10-15min) to collect the core-shell carrier matrix;

[0010] (2) Strong force unit construction and drug loading: dissolve the drug active ingredient (concentration 0.5-2 mg / mL), the main force reagent (molar ratio 1:1-1:3 with the drug), in the buffer (pH 5.5-7.5), add the core-shell carrier matrix (mass ratio of drug to carrier 1:5-1:20), react at 25-35℃ for 4-6h, then add the auxiliary force reagent (concentration 0.1-1 mg / mL), continue to react for 2-3h, to form a drug-carrier-strong force complex;

[0011] (3) Surface functional modification: add the targeting ligand (mass ratio 1:10-1:20 with the carrier) and the PEGylation reagent (molecular weight 2000-10000 Da, modification density 0.5-2 μg / cm 2 ) to the above complex, react at 37℃ for 3-5h, to achieve "targeting-anti-clearance" dual functional modification;

[0012] (4) Purification and quality control: purify by gel filtration chromatography (eluent PBS, flow rate 1-2 mL / min), collect the target component, measure the particle size (80-400 nm), PDI (≤0.25), drug encapsulation efficiency (≥70%) and stimulus-responsive release efficiency (24h cumulative release rate ≥85% under double stimuli), to obtain the intelligent drug delivery system.

[0013] As an improvement, the mesoporous core layer of the core-shell structure carrier matrix is selected from one of mesoporous silica, mesoporous hydroxyapatite, metal organic framework material (MOFs, such as ZIF-8, UiO-66); the shell polymer is selected from at least one of polylactic acid-glycolic acid copolymer (PLGA, lactic acid / glycolic acid molar ratio 50:50-75:25), polyethylene glycol-poly (ε-caprolactone) block copolymer (PEG-PCL, PEG content 10%-30%), chitosan quaternary ammonium salt (substitution degree ≥60%), and the shell polymer needs to be grafted with active groups (amino, carboxyl or mercapto) capable of reacting with the main force reagent.

[0014] As an improvement, in the strong force unit:

[0015] When the main force is coordination bond, it is formed by metal ions (Fe 3+ , Zn 2+ or Cu 2+ , concentration 0.1-1 mmol / L) and coordination groups (hydroxyl, amino, carboxyl or o-phenanthroline groups) in the carrier shell / drug active ingredient, with a stability constant Kf of 10 8 -10 12 L / mol, and Kf decreases to 10 3 -10 5 L / mol in an acidic environment at pH 4.0-5.0;

[0016] when the main force is dynamic covalent bond, selected from Schiff base bond (formed by reaction of aldehyde-modified carrier with amino-modified drug, broken at pH 4.0-5.0 or glutathione concentration ≥10 mmol / L), disulfide bond (formed by reaction of thiol-modified drug with disulfide succinimide ester modified carrier, broken at glutathione concentration ≥5 mmol / L or in the presence of matrix metalloproteinase-2 / 9);

[0017] when the auxiliary force is hydrogen bond, formed by hydroxyl / amino of carrier shell and carbonyl / hydroxyl of drug, bond energy is 2-8 kJ / mol; when the auxiliary force is π-π stacking, formed by aromatic ring (such as benzene ring, naphthalene ring) of carrier surface and aromatic ring of drug, bond energy is 1-5 kJ / mol.

[0018] As an improvement, the dual physiological microenvironment stimuli are selected from one of the following combinations: ① pH 4.5-5.5 acidic environment + glutathione concentration 10-20 mmol / L; ② temperature 39-42℃ + matrix metalloproteinase-2 / 9 (concentration ≥10 ng / mL); ③ pH 4.0-5.0 acidic environment + cysteine protease (concentration ≥5 ng / mL); and the trigger delay time of stimuli response is ≤30 min, and the response completion time is ≤2 h.

[0019] As an improvement, the surface functionalization layer comprises a targeting ligand and an anti-clearance modification; the targeting ligand is selected from at least one of folate (modification density 20-40 / μm 2 , binding constant for folate receptor positive cells ≥10 9 L / mol), RGD peptide (sequence Arg-Gly-Asp, modification density 15-35 / μm 2 , targeting efficiency for integrin αvβ3 ≥80%), transferrin (modification density 5-15 / μm 2 , targeting efficiency for transferrin receptor ≥75%); the anti-clearance modification is PEGylation modification (molecular weight 5000-8000 Da, modification density 1-1.5 μg / cm 2 ), and the PEG terminal can be grafted with a fluorescent probe (such as FITC, Cy5) for in vivo tracking.

[0020] As an improvement, the pharmaceutically active ingredient is selected from one of the following combinations: ① small molecule chemotherapeutic drugs (doxorubicin, paclitaxel or cisplatin, loading capacity 8%-15% mass fraction) + photosensitizer (hematoporphyrin monomethyl ether, loading capacity 2%-5% mass fraction); ② antibody drugs (trastuzumab, bevacizumab, loading capacity 3%-8% mass fraction) + small molecule anti-inflammatory drugs (ibuprofen, loading capacity 5%-10% mass fraction); ③ nucleic acid drugs (siRNA, miRNA or plasmid DNA, loading capacity 1%-3% mass fraction) + small molecule chemotherapeutic drugs (cisplatin, loading capacity 5%-12% mass fraction); and the dispersion uniformity of the pharmaceutically active ingredient in the delivery system is ≥ 90%, without obvious agglomeration (agglomerated particles ≤ 5%).

[0021] As an improvement, in step (1), if the mesoporous core layer is metal organic framework material ZIF-8, the preparation process is: mixing zinc nitrate (concentration 0.1-0.5 mol / L) and 2-methyl imidazole (concentration 0.4-2 mol / L) at a molar ratio of 1:4-1:8, stirring at 25-30℃ in methanol solvent for 2-4 h, washing 3-5 times with methanol after centrifugation (10000 rpm, 10 min), and vacuum drying (60℃, 12 h) to obtain ZIF-8 mesoporous core layer; if the shell layer is PLGA, the polymer solution is prepared using dichloromethane / acetone mixed solvent (volume ratio 3:1-5:1), and the crosslinking agent is selected from a mixture of 1-ethyl-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC) and N-hydroxysuccinimide (NHS) (molar ratio 1:1).

[0022] As an improvement, in step (2), if the main force is Schiff base bond, the pharmaceutically active ingredient is aminyl doxorubicin (prepared by reacting doxorubicin with ethylenediamine at a molar ratio of 1:2-1:5), and the carrier shell layer is aldehyde PLGA (prepared by reacting PLGA with glutaraldehyde at a mass ratio of 10:1-20:1), and the reaction is carried out at pH 6.8-7.2 and temperature 32-35℃ for 5-6 h, and then an auxiliary force reagent (such as catechol, concentration 0.5 mg / mL) is added and reacted for 2.5 h to form a Schiff base bond-hydrogen bond synergistically mediated complex; in step (3), the PEGylation reagent is selected from methoxypolyethylene glycol-succinimidyl carbonate (mPEG-SC), and the reaction with the carrier is carried out in PBS buffer at pH 7.2-7.4, and after the reaction is completed, the particle size change is monitored in real time by dynamic light scattering instrument to ensure that the particle size increase after PEGylation modification is ≤ 20 nm.

[0023] As an improvement, in the preparation of drugs for the treatment of specific subtype diseases, specifically: ① for HER2 positive breast cancer, select RGD peptide-PEG double modification, Schiff base bond-π-π stacking mediated delivery system, load trastuzumab and paclitaxel, and release by double stimulus response of pH5.0-5.5+ glutathione 15mmol / L; ② for ulcerative colitis, select folic acid-PEG double modification, disulfide bond-hydrogen bond mediated delivery system, load ibuprofen and siRNA (targeting TNF-α gene), and release by double stimulus response of pH4.5-5.0+ cysteine protease; ③ for non-alcoholic fatty liver, select galactose-PEG double modification, coordination bond (Zn 2+ )-hydrogen bond mediated delivery system, load plasmid DNA (targeting PPAR-γ gene) and curcumin, and release by double stimulus response of temperature 40-42℃+pH5.0-5.5; and the enrichment amount of the delivery system in the target organ is ≥30% of the injection dose, and the distribution amount in the non-target organ is ≤10% of the injection dose.

[0024] The advantages of the present application compared with the prior art are that the technical solution has the following advantages: first, the strong force unit of "main force + auxiliary force" is combined with the mesoporous structure of the core-shell carrier to make the drug loading more stable, the 24h leakage rate is only 5%, the encapsulation rate is more than 75%, and the drug is more economical and less toxic than the traditional system. Second, the double stimulus response mechanism is designed to match the environment of different disease target sites to achieve "pulse-slow release", 30%-50% of the drug is released within 1h to reach the treatment threshold, and the subsequent 23h slow release is 40%-60%, which prolongs the efficacy. Third, the carrier surface is modified with targeting ligands and PEG, which has strong targeting ability, high enrichment in tumor, colon and other target sites, and less distribution in non-target organs, and PEG can prolong the circulation time in the body and improve the bioavailability. Fourth, the adjustable components can be adapted to various diseases, the carrier is made of safe materials, and the preparation has no toxic residues, so that the drug is safer. Fifth, the preparation process parameters are clear, common equipment is used, quality control meets the standards, easy to scale up production, and meets the requirements of clinical industrialization. BRIEF DESCRIPTION OF DRAWINGS

[0025] Figure 1 is the system core structure and release mechanism flow chart of the strong force mediated intelligent drug delivery system and preparation method of the present application.

[0026] Figure 2 is the preparation process flow of the strong force mediated intelligent drug delivery system and preparation method of the present application. DETAILED DESCRIPTION

[0027] For the purpose of promoting an understanding of the application, the application will now be described in greater detail with reference to the figures. The embodiments shown in the figures are intended to explain the disclosure and to provide a basis for a working of the application, but they do not limit the scope of the application. Various embodiments of the application will be presented in the following description with reference to the attached drawings. The figures are provided to illustrate the application and to provide a basis for a working of the application, but they do not limit the scope of the application. Various embodiments of the application will be presented in the following description with reference to the attached drawings.

[0028] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs. The terminology used in the description of the application herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the application.

[0029] It will be understood that the spatially relative terms "beneath", "below", "lower", "under", "above", "upper" and the like, can be used herein for ease of description to describe one element or feature's relationship to another element(s) or feature(s) as illustrated in the figures. It will be understood that the spatially relative terms are intended to encompass different orientations of the device in use or operation in addition to the orientations depicted in the figures. For example, if the device in the figures is turned over, elements described as "below" or "beneath" other elements or features would then be oriented "above" the other elements or features. Thus, the exemplary term "below" can encompass both an orientation of above and below. The device can also be oriented in the other directions, such as rotated 90 degrees or at other orientations, and the spatially relative terms used herein interpreted accordingly.

[0030] It is to be noted that when an element is referred to as being "connected" to another element, it can be directly connected to the other element, or connected to the other element with intervening elements. "Connected" in the following embodiments should be understood as "electrically connected", "communicatively connected" and the like if the circuits, modules, units and the like connected to each other have transmission of electrical signals or data.

[0031] As used herein, the singular forms "a", "an" and "the" include plural referents unless the context clearly dictates otherwise. It will be further understood that the terms "comprises", "comprising", "includes" and / or "including", or the like, when used in this specification, specify the presence of stated features, integers, steps, operations, elements, components, or combinations thereof, but do not preclude the presence or addition of one or more other features, integers, steps, operations, elements, components, or combinations thereof.

[0032] In combination with the drawings, a strong force mediated intelligent drug delivery system and a preparation method, comprising a core-shell structure carrier matrix, a drug active ingredient, a strong force unit mediating specific binding of the two, and a surface functionalization layer; the core layer of the core-shell structure carrier matrix is a mesoporous material (pore size 2-10 nm), the shell layer is a biocompatible polymer coating (thickness 10-50 nm), the overall particle size is 80-400 nm and the dispersion coefficient PDI is ≤0.25; the strong force unit is composed of "main force + auxiliary force", the main force is selected from one of coordination bond and dynamic covalent bond, the auxiliary force is selected from at least one of hydrogen bond and π-π stacking interaction, and the main force can reversibly break / weaken in response to double physiological microenvironment stimuli, and the auxiliary force cooperatively controls the drug release kinetics; under the stimulation of the double physiological microenvironment, the main force breaks to make the drug-carrier binding force decrease by ≥60%, the auxiliary force maintains the stability of the carrier structure, and realizes the "pulse-slow release" dual-mode release of the drug active ingredient at the target site (24h cumulative release rate: 30%-50% released within 1h in the pulse stage, and 40%-60% released within 23h in the slow release stage).

[0033] As an improvement, the following steps are included:

[0034] (1) Core-shell carrier matrix preparation: ① Mesoporous core layer preparation: using tetraethyl orthosilicate / metal organic framework material as raw material, cetyltrimethylammonium bromide / polyetheramine as template agent, sol-gel reaction at pH 8.5-10.0 and temperature 45-60℃ for 8-12h, calcination (500-600℃, 4-6h) or solvent extraction to remove the template agent, to obtain the mesoporous core layer; ② Shell coating: dispersing the mesoporous core layer in a polymer solution (concentration 1-5mg / mL), under nitrogen protection and stirring rate 300-500rpm, adding crosslinking agent (mass ratio to polymer 1:5-1:10), reacting at 30-40℃ for 6-8h, centrifuging (8000-12000rpm, 10-15min) to collect the core-shell carrier matrix;

[0035] (2) Strong force unit construction and drug loading: dissolving the drug active ingredient (concentration 0.5-2mg / mL) and the main force reagent (molar ratio to drug 1:1-1:3) in buffer (pH 5.5-7.5), adding the core-shell carrier matrix (mass ratio of drug to carrier 1:5-1:20), reacting at 25-35℃ for 4-6h, then adding the auxiliary force reagent (concentration 0.1-1mg / mL), continuing to react for 2-3h, to form a drug-carrier-strong force complex;

[0036] (3) Surface functionalization modification: adding targeting ligand (1:10-1:20 with the mass of carrier) and PEGylation reagent (molecular weight 2000-10000 Da, modification density 0.5-2 μg / cm 2 ) to the above complex, reacting at 37℃ for 3-5 h to achieve "targeting-anti-clearance" bifunctional modification;

[0037] (4) Purification and quality control: purification by gel filtration chromatography (eluent PBS, flow rate 1-2 mL / min), collection of target components, determination of particle size (80-400 nm), PDI (≤0.25), drug encapsulation efficiency (≥70%) and stimulus-responsive release efficiency (24 h cumulative release rate ≥85% under double stimuli) to obtain the intelligent drug delivery system.

[0038] As an improvement, the mesoporous core layer of the core-shell structure carrier matrix is selected from one of mesoporous silica, mesoporous hydroxyapatite, metal organic framework material (MOFs, such as ZIF-8, UiO-66); the shell polymer is selected from at least one of polylactic acid-glycolic acid copolymer (PLGA, lactic acid / hydroxyacetic acid molar ratio 50:50-75:25), polyethylene glycol-poly (ε-caprolactone) block copolymer (PEG-PCL, PEG content 10%-30%), chitosan quaternary ammonium salt (substitution degree ≥60%), and the shell polymer needs to be grafted with active groups (amino, carboxyl or sulfhydryl) capable of reacting with the main force reagent.

[0039] As an improvement, in the strong force unit:

[0040] When the main force is coordination bond, it is formed by metal ions (Fe 3+ , Zn 2+ or Cu 2+ , concentration 0.1-1 mmol / L) and coordination groups (hydroxyl, amino, carboxyl or o-phenanthroline groups) in the carrier shell / drug active ingredient, with a stability constant Kf of 10 8 -10 12 L / mol, and Kf decreases to 10 3 -10 5 L / mol in a pH 4.0-5.0 acidic environment;

[0041] When the main force is dynamic covalent bond, it is selected from Schiff base bond (formed by reacting aldehyde-modified carrier with amino-modified drug, broken at pH 4.0-5.0 or glutathione concentration ≥10 mmol / L), disulfide bond (formed by reacting sulfhydryl-modified drug with disulfide succinimide ester-modified carrier, broken at glutathione concentration ≥5 mmol / L or in the presence of matrix metalloproteinase-2 / 9);

[0042] When the auxiliary force is hydrogen bond, it is formed by the hydroxyl / amino group of the carrier shell and the carbonyl / hydroxyl group of the drug, and the bond energy is 2-8 kJ / mol; when the auxiliary force is π-π stacking, it is formed by the aromatic ring (such as benzene ring, naphthalene ring) on the surface of the carrier and the aromatic ring of the drug, and the interaction energy is 1-5 kJ / mol.

[0043] As an improvement, the double physiological microenvironment stimulation is selected from one of the following combinations: ① pH 4.5-5.5 acidic environment + glutathione concentration 10-20 mmol / L; ② temperature 39-42℃ + matrix metalloproteinase-2 / 9 (concentration ≥10 ng / mL); ③ pH 4.0-5.0 acidic environment + cysteine protease (concentration ≥5 ng / mL); and the trigger delay time of the stimulation response is ≤30 min, and the response completion time is ≤2 h.

[0044] As an improvement, the surface functionalization layer comprises a targeting ligand and an anti-clearance modification; the targeting ligand is selected from at least one of folate (modification density 20-40 / μm 2 , binding constant to folate receptor positive cells ≥10 9 L / mol), RGD peptide (sequence Arg-Gly-Asp, modification density 15-35 / μm 2 , binding rate to integrin αvβ3 ≥80%), transferrin (modification density 5-15 / μm 2 , targeting efficiency to transferrin receptor ≥75%); the anti-clearance modification is PEGylation modification (molecular weight 5000-8000 Da, modification density 1-1.5 μg / cm 2 ), and the PEG terminal can be grafted with a fluorescent probe (such as FITC, Cy5) for in vivo tracing.

[0045] As an improvement, the pharmaceutically active ingredient is selected from one of the following combinations: ① small molecule chemotherapeutic drug (doxorubicin, paclitaxel or cisplatin, loading amount 8%-15% mass fraction) + photosensitizer (monomethyl ether of hematoporphyrin, loading amount 2%-5% mass fraction); ② antibody drug (trastuzumab, bevacizumab, loading amount 3%-8% mass fraction) + small molecule anti-inflammatory drug (ibuprofen, loading amount 5%-10% mass fraction); ③ nucleic acid drug (siRNA, miRNA or plasmid DNA, loading amount 1%-3% mass fraction) + small molecule chemotherapeutic drug (cisplatin, loading amount 5%-12% mass fraction); and the dispersion uniformity of the pharmaceutically active ingredient in the delivery system is ≥90%, and there is no obvious aggregation (aggregated particles account for ≤5%).

[0046] As an improvement, in step (1), if the mesoporous core layer is metal-organic framework material ZIF-8, the preparation process is as follows: zinc nitrate (concentration 0.1-0.5 mol / L) and 2-methyl imidazole (concentration 0.4-2 mol / L) are mixed in a molar ratio of 1:4-1:8, stirred at 25-30℃ in methanol solvent for 2-4 h, washed with methanol 3-5 times after centrifugation (10000 rpm, 10 min), and vacuum dried (60℃, 12 h) to obtain ZIF-8 mesoporous core layer; if the shell layer is PLGA, the polymer solution is prepared using dichloromethane / acetone mixed solvent (volume ratio 3:1-5:1), and the crosslinking agent is a mixture of 1-ethyl-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC) and N-hydroxysuccinimide (NHS) (molar ratio 1:1).

[0047] As an improvement, in step (2), if the main force is Schiff base bond, the active pharmaceutical ingredient is aminyl doxorubicin (prepared by reacting doxorubicin with ethylenediamine in a molar ratio of 1:2-1:5), and the carrier shell layer is aldehyde PLGA (prepared by reacting PLGA with glutaraldehyde in a mass ratio of 10:1-20:1), the reaction is carried out at pH 6.8-7.2 and temperature 32-35℃ for 5-6 h, then an auxiliary force reagent (such as catechol, concentration 0.5 mg / mL) is added and reacted for 2.5 h to form a Schiff base bond-hydrogen bond synergistically mediated complex; in step (3), the PEGylation reagent is selected to be methoxy polyethylene glycol-succinimidyl carbonate (mPEG-SC), and the reaction with the carrier is carried out in PBS buffer at pH 7.2-7.4, and after the reaction is completed, the particle size change is monitored in real time by dynamic light scattering instrument to ensure that the particle size increase after PEGylation modification is ≤20 nm.

[0048] As an improvement, in the application for preparing a drug for treating a specific subtype disease, specifically: ① for HER2-positive breast cancer, a RGD peptide-PEG double-modified, Schiff base bond-π-π stacking mediated delivery system is selected, loading trastuzumab and paclitaxel, and releasing by double stimulus response of pH 5.0-5.5+ glutathione 15 mmol / L; ② for ulcerative colitis, a folic acid-PEG double-modified, disulfide bond-hydrogen bond mediated delivery system is selected, loading ibuprofen and siRNA (targeting TNF-α gene), and releasing by double stimulus response of pH 4.5-5.0+ cysteine protease; ③ for non-alcoholic fatty liver, a galactose-PEG double-modified, coordination bond (Zn 2+ )-hydrogen bond mediated delivery system is selected, loading plasmid DNA (targeting PPAR-γ gene) and curcumin, and releasing by double stimulus response of temperature 40-42℃+ pH 5.0-5.5; and the enrichment amount of the delivery system in the target organ is ≥30% of the injection dose, and the distribution amount in the non-target organ is ≤10% of the injection dose.

[0049] I. Example 1: Intelligent drug delivery system for the treatment of HER2-positive breast cancer:

[0050] (I) Preparation of raw materials:

[0051] Mesoporous core layer raw materials: tetraethyl orthosilicate (analytical pure, Sinopharm Group), cetyltrimethylammonium bromide (CTAB, purity ≥ 99%, Sigma-Aldrich), ammonia water (25% by mass, Sinopharm Group);

[0052] Shell polymer raw materials: poly(lactic-co-glycolic acid) (PLGA, lactic acid / glycolic acid molar ratio 75:25, molecular weight 50000 Da, Sigma-Aldrich), glutaraldehyde (50% by mass, Sinopharm Group), 1-ethyl-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC, purity ≥ 98%, Aladdin), N-hydroxysuccinimide (NHS, purity ≥ 98%, Aladdin);

[0053] Pharmaceutically active ingredients: doxorubicin (purity ≥ 99%, Sigma-Aldrich), ethylenediamine (analytical pure, Sinopharm Group), trastuzumab (purity ≥ 95%, Roche), hematoporphyrin monomethyl ether (purity ≥ 98%, Sigma-Aldrich);

[0054] Strong force unit reagent: catechol (purity ≥ 99%, Sigma-Aldrich);

[0055] Surface functionalization reagent: RGD peptide (sequence Arg-Gly-Asp, purity ≥ 98%, Jier Biochemical), methoxy polyethylene glycol-succinimidyl carbonate (mPEG-SC, molecular weight 5000 Da, purity ≥ 95%, Sigma-Aldrich);

[0056] Buffer and solvent: phosphate buffer (PBS, pH 7.4, 0.01 mol / L), dichloromethane (analytical pure, Sinopharm Group), acetone (analytical pure, Sinopharm Group), methanol (analytical pure, Sinopharm Group).

[0057] (II) Preparation steps:

[0058] Preparation of aminated doxorubicin: dissolve doxorubicin (0.1 g) in methanol (10 mL), add ethylenediamine (0.05 mL, doxorubicin to ethylenediamine molar ratio 1:3), stir at 30°C for 4 h, remove methanol by rotary evaporation, vacuum dry (60°C, 8 h) to obtain aminated doxorubicin;

[0059] Mesoporous silica core layer preparation: add deionized water (50 mL), CTAB (0.5 g) into a three-necked flask, stir at 50 °C until completely dissolved, add ammonia water (2 mL) to adjust pH to 9.0, slowly drop tetraethyl orthosilicate (2 mL), incubate at 50 °C for 10 h; after the reaction is completed, centrifuge (10000 rpm, 15 min) to collect the precipitate, wash with deionized water for 3 times, calcine at 550 °C for 5 h to remove CTAB, and obtain mesoporous silica core layer (pore size 5-8 nm);

[0060] Aldehyde-modified PLGA shell layer coating: dissolve PLGA (1 g) in a dichloromethane / acetone mixed solvent (volume ratio 4:1, 20 mL), add glutaraldehyde (0.1 g, PLGA to glutaraldehyde mass ratio 10:1), stir at 35 °C for 3 h to obtain an aldehyde-modified PLGA solution; disperse the mesoporous silica core layer (0.2 g) in the above solution, add an EDC / NHS mixture (molar ratio 1:1, total mass 0.1 g) under nitrogen protection, stir at 35 °C for 7 h; centrifuge (10000 rpm, 12 min) to collect the precipitate, wash with dichloromethane for 2 times, and vacuum dry (40 °C, 12 h) to obtain an aldehyde-modified PLGA-coated core-shell carrier matrix (particle size 120-150 nm, PDI=0.18);

[0061] Strong force unit construction and drug loading: dissolve aminated doxorubicin (0.02 g, concentration 1 mg / mL), trastuzumab (0.01 g), and hematoporphyrin monomethyl ether (0.005 g) in PBS (pH 6.8, 20 mL), add the core-shell carrier matrix (0.2 g, drug to carrier mass ratio 1:10), stir at 32 °C for 5 h; add catechol (0.005 g, concentration 0.25 mg / mL), continue to react for 2.5 h to form a Schiff base bond-hydrogen bond synergistically mediated drug-carrier complex;

[0062] Surface functionalization modification: add RGD peptide (0.02 g, mass ratio to carrier 1:10), mPEG-SC (0.03 g, modification density 1.2 μg / cm 2 ), stir at 37 °C for 4 h;

[0063] Purification and quality control: purify by gel filtration chromatography (eluent PBS, flow rate 1.5 mL / min), and collect the target component; the particle size is 130-160 nm (the particle size increases by ≤15 nm after PEGylation modification), PDI=0.20, the drug encapsulation rate is 82%, the cumulative release rate under double stimulation (pH 5.2+glutathione 15 mmol / L) for 24 h is 90% (42% released in the pulse stage for 1 h, and 48% released in the slow-release stage for 23 h), and an intelligent drug delivery system for HER2-positive breast cancer treatment is obtained.

[0064] (III) Performance test and application effect:

[0065] Targeting test: flow cytometry was used to detect the uptake rate of the delivery system by HER2-positive breast cancer cells (SK-BR-3 cells), and the results showed that the uptake rate reached 85%, which was significantly higher than that of the non-targeted delivery system (30%);

[0066] In vivo distribution test: the fluorescently labeled delivery system was injected into SK-BR-3 tumor-bearing mice through the tail vein, and the fluorescence intensity at the tumor site was 3.5 times that of the liver after 24 hours of dissection detection, and the tumor enrichment amount reached 35% of the injection dose, and the distribution amount in non-target organs (heart, kidney) was ≤8%;

[0067] Tumor inhibition effect test: SK-BR-3 tumor-bearing mice were injected with drugs through the tail vein (dose 10 mg / kg, once every 3 days, a total of 5 times), and after 21 days of treatment, the tumor volume inhibition rate reached 78%, and the body weight of the mice did not decrease significantly (body weight change rate ≤5%).

[0068] II. Example 2: Intelligent drug delivery system for the treatment of ulcerative colitis:

[0069] (I) Raw material preparation:

[0070] Mesoporous core layer raw material: metal organic framework material ZIF-8 precursor (zinc nitrate, purity ≥ 99%; 2-methylimidazole, purity ≥ 99%, both from Sigma-Aldrich);

[0071] Shell polymer raw material: chitosan quaternary ammonium salt (degree of substitution 70%, molecular weight 30000 Da, Sigma-Aldrich), EDC, NHS;

[0072] Pharmaceutically active ingredients: ibuprofen (purity ≥ 99%, Sigma-Aldrich), siRNA (targeting TNF-α gene, purity ≥ 98%, Genomed);

[0073] Strong force unit reagent: dithiothreitol ester (purity ≥ 98%, Aladdin), mercaptoacetic acid (analytical pure, Sinopharm Group);

[0074] Surface functionalization reagent: folic acid (purity ≥ 99%, Sigma-Aldrich), mPEG-SC (molecular weight 8000 Da, Sigma-Aldrich);

[0075] Buffer and solvent: PBS (pH 7.2, 0.01 mol / L), methanol, deionized water.

[0076] (II) Preparation steps:

[0077] Preparation of sulfhydrylated ibuprofen: ibuprofen (0.1 g) was dissolved in methanol (10 mL), and mercaptoacetic acid (0.03 mL, ibuprofen to mercaptoacetic acid molar ratio 1:2) was added. The reaction was stirred at 35 °C for 5 h. Methanol was removed by rotary evaporation, and the product was dried in vacuum (50 °C, 6 h) to obtain sulfhydrylated ibuprofen;

[0078] Preparation of ZIF-8 mesoporous core layer: zinc nitrate (0.2 g, concentration 0.3 mol / L) was dissolved in methanol (10 mL), and 2-methylimidazole (0.6 g, concentration 1.2 mol / L) was dissolved in methanol (10 mL). The two solutions were mixed (zinc nitrate to 2-methylimidazole molar ratio 1:6), and the reaction was stirred at 28 °C for 3 h. The precipitate was collected by centrifugation (10000 rpm, 10 min), washed with methanol 4 times, and dried in vacuum (60 °C, 12 h) to obtain ZIF-8 mesoporous core layer (pore size 3-5 nm);

[0079] Coating of dithio succinimide ester modified chitosan quaternary ammonium salt shell layer: chitosan quaternary ammonium salt (0.8 g) was dissolved in PBS (pH 7.2, 20 mL), and dithio succinimide ester (0.08 g, polymer to dithio succinimide ester mass ratio 1:10) was added. The reaction was stirred at 30 °C for 4 h. ZIF-8 core layer (0.16 g) was added, and EDC / NHS mixture (molar ratio 1:1, total mass 0.08 g) was added. The reaction was stirred at 30 °C for 6 h. The precipitate was collected by centrifugation (9000 rpm, 10 min), washed with PBS 2 times, and dithio succinimide ester modified core-shell carrier matrix (particle size 90-120 nm, PDI = 0.15) was obtained;

[0080] Construction of strong force unit and drug loading: sulfhydrylated ibuprofen (0.016 g, concentration 0.8 mg / mL) and siRNA (0.003 g, concentration 0.15 mg / mL) were dissolved in PBS (pH 7.0, 20 mL), and core-shell carrier matrix (0.16 g, drug to carrier mass ratio 1:8) was added. The reaction was stirred at 30 °C for 5 h to form dithio bond-hydrogen bond synergistically mediated drug-carrier complex;

[0081] Surface functionalization modification: folic acid (0.016 g, carrier to folic acid mass ratio 1:10) and mPEG-SC (0.024 g, modification density 1.5 μg / cm 2 ) were added, and the reaction was stirred at 37 °C for 3.5 h;

[0082] Purification and quality control: gel filtration chromatography purification (eluent PBS, flow rate 1.2 mL / min), target components were collected; particle size 100-130 nm, PDI = 0.18, drug encapsulation rate 78%, 24h cumulative release rate 88% under double stimulation (pH 4.8 + cysteine protease 5ng / mL) (pulse phase 1h release 38%, slow release phase 23h release 50%), to obtain an intelligent drug delivery system for the treatment of ulcerative colitis.

[0083] (III) Performance test and application effect:

[0084] Intestinal targeting test: ulcerative colitis model rats were given intragastrically, and the drug concentration in different parts of the intestine was detected after 24h. The drug concentration in the colon was 8 times that in the stomach and 4 times that in the small intestine, and the colon targeting efficiency reached 82%;

[0085] Anti-inflammatory effect test: after 7 days of administration, the expression level of TNF-α mRNA in the colon tissue of rats was detected, and the results showed that the expression amount decreased by 65% compared with the model group, and the colon mucosa damage score decreased from 4 points (severe damage) to 1 point (mild damage);

[0086] Safety test: after 14 days of continuous administration, there was no obvious abnormality in the liver and kidney function indicators (ALT, AST, BUN, Cr) of rats, and there was no obvious irritation reaction in the colon mucosa.

[0087] III. Example 3: Intelligent drug delivery system for the treatment of non-alcoholic fatty liver disease:

[0088] (I) Raw material preparation:

[0089] Mesoporous core layer raw material: mesoporous hydroxyapatite precursor (calcium nitrate, diammonium hydrogen phosphate, both analytical pure, Sinopharm Group);

[0090] Shell polymer raw material: polyethylene glycol-poly (caprolactone) block copolymer (PEG-PCL, PEG content 20%, molecular weight 40000 Da, Sigma-Aldrich), EDC, NHS;

[0091] Drug active ingredient: curcumin (purity ≥98%, Sigma-Aldrich), plasmid DNA (targeting PPAR-γ gene, purity ≥95%, Genmark);

[0092] Strong force unit reagent: zinc nitrate (purity ≥99%, Sigma-Aldrich);

[0093] Surface functionalization reagent: galactose (purity ≥99%, Sigma-Aldrich), mPEG-SC (molecular weight 6000 Da, Sigma-Aldrich);

[0094] Buffer and solvent: PBS (pH 7.4, 0.01 mol / L), deionized water, ethanol (analytical pure, National Pharmaceutical Group).

[0095] (ii) Preparation step:

[0096] Preparation of mesoporous hydroxyapatite core layer: calcium nitrate (0.5 g) was dissolved in deionized water (20 mL), diammonium hydrogen phosphate (0.3 g) was dissolved in deionized water (20 mL), the two solutions were mixed (Ca / P molar ratio 1.67), the pH was adjusted to 10.0 with ammonia water, and the mixture was stirred at 60°C for 12 h; the precipitate was collected by centrifugation (8000 rpm, 15 min) and washed with deionized water 3 times, and then calcined at 800°C for 4 h to obtain mesoporous hydroxyapatite core layer (pore size 4-6 nm);

[0097] PEG-PCL shell coating: PEG-PCL (1 g) was dissolved in ethanol (20 mL), mesoporous hydroxyapatite core layer (0.2 g) was added, EDC / NHS mixture (molar ratio 1:1, total mass 0.1 g) was added, and the mixture was stirred at 35°C for 8 h; the precipitate was collected by centrifugation (9000 rpm, 12 min) and washed with ethanol 2 times, and then vacuum dried (50°C, 10 h) to obtain PEG-PCL coated core-shell carrier matrix (particle size 150-180 nm, PDI = 0.22);

[0098] Strong force unit construction and drug loading: curcumin (0.02 g, concentration 1 mg / mL) and plasmid DNA (0.004 g, concentration 0.2 mg / mL) were dissolved in PBS (pH 7.4, 20 mL), zinc nitrate solution (0.005 g, concentration 0.25 mmol / L) was added, and the mixture was stirred for 30 min; core-shell carrier matrix (0.2 g, drug to carrier mass ratio 1:10) was added, and the mixture was stirred at 35°C for 6 h to form Zn 2+ coordination bond-hydrogen bond synergistically mediated drug-carrier complex;

[0099] Surface functionalization modification: galactose (0.02 g, mass ratio to carrier 1:10) and mPEG-SC (0.03 g, modification density 1.3 μg / cm 2 ) were added, and the mixture was stirred at 37°C for 4.5 h;

[0100] Purification and quality control: gel filtration chromatography purification (eluent PBS, flow rate 1.8 mL / min), and the target component was collected; the particle size was 160-190 nm, PDI = 0.23, drug encapsulation efficiency was 75%, and the cumulative release rate under double stimulation (temperature 41°C + pH 5.2) for 24 h was 86% (pulse phase 1 h release 35%, slow release phase 23 h release 51%), thus obtaining an intelligent drug delivery system for the treatment of non-alcoholic fatty liver.

[0101] (III) Performance testing and application effect:

[0102] Hepatocyte targeting test: fluorescence microscopy was used to observe the uptake of the delivery system by human hepatocytes (HepG2 cells), and the results showed that the uptake rate was 80%, which was significantly higher than that of the non-targeting group (25%);

[0103] In vivo efficacy test: high-fat diet-induced non-alcoholic fatty liver mice were injected with drugs through the tail vein (dose 8 mg / kg, twice a week, for a total of 4 weeks), and liver function indicators were detected. The levels of ALT and AST were reduced by 45% and 40%, respectively, and the liver triglyceride content was reduced by 50%;

[0104] Histopathological examination: after 4 weeks of administration, liver tissue sections of mice showed that the degree of hepatocyte steatosis was significantly reduced, the infiltration of inflammatory cells was reduced, and the liver lobule structure was basically restored to normal.

[0105] Four, common test method explanation:

[0106] Particle size and PDI determination: dynamic light scattering instrument (Malvern Zetasizer NanoZS90) was used, PBS was used as the dispersion medium, and the determination was carried out at room temperature, 3 times for each sample, and the average value was taken;

[0107] Nucleic acid drug encapsulation rate determination: high salt solution (such as 1M NaCl) or nuclease-assisted demulsification was used, and the amount of free nucleic acid was quantified by agarose gel electrophoresis, and the encapsulation rate was calculated.

[0108] Stimulus-responsive release test: dialysis bag method (molecular weight cutoff 10000 Da) was used, and samples were taken at different times under simulated normal physiological environment (pH 7.4, 37℃) and double stimulation environment, respectively. HPLC was used to determine the drug release amount, and the cumulative release rate was calculated;

[0109] In vivo distribution test: the delivery system was labeled with FITC fluorescence, and was injected into animal models through the tail vein. Organs were dissected at different time points, and the fluorescence intensity of each organ was measured by fluorescence spectrophotometer (Shimadzu RF-6000), and the drug distribution amount was calculated.

[0110] Low utilization degree and other technical bottlenecks, which have significant advantages in drug delivery efficiency, treatment safety, clinical applicability and industrialization application, as follows:

[0111] Strong force mediated to improve drug loading stability and delivery efficiency: dual force synergistically enhances loading stability: compared with traditional systems with single force (such as pure hydrogen bond, physical adsorption), the present scheme uses strong force units (such as Schiff base bond-hydrogen bond, disulfide bond-hydrogen bond, Zn2+ Coordination bond-hydrogen bond), main acting force (stability constant 10 8 -10 12 L / mol) ensure high strength binding of drugs and carriers, auxiliary force (bond energy 1-8 kJ / mol) further optimize the stability of the binding, effectively solve the problem of "leakage in advance" of the drug in the process of in vivo transport. As in Example 1, the Schiff base bond-hydrogen bond mediated system has only 5% drug leakage rate in 24h under normal physiological environment (pH 7.4, 37℃), which is much lower than that of traditional physical adsorption system (leakage rate ≥20%).

[0112] High encapsulation efficiency reduces drug waste and side effects: through the specific binding of strong force units and the mesoporous structure of the core-shell carrier matrix (pore size 2-10 nm), efficient loading of different types of drugs such as small molecule chemotherapeutic drugs, antibody drugs, and nucleic acid drugs is achieved, with drug encapsulation efficiency generally ≥75% (Example 1 82%, Example 2 78%, Example 3 75%), which is significantly higher than that of traditional liposome systems (encapsulation efficiency usually 50%-70%). High encapsulation efficiency not only reduces drug raw material waste, but also reduces the toxic damage of non-loaded free drugs to non-target tissues, making low-dose high-efficiency treatment possible in clinical practice.

[0113] Intelligent dual response for precise and controllable drug release: Dual stimulus response adapts to complex physiological microenvironment: In view of the specific physiological microenvironment of different disease target sites (such as the acidity + high glutathione of tumor tissue, the acidity + high enzyme concentration of inflammatory intestinal tract, and the warm + acidity of fatty liver tissue), the scheme designs "pH-glutathione" "pH-enzyme" "temperature-pH" and other dual stimulus response mechanisms to trigger conditions that precisely match pathological environments (such as tumor pH 4.5-5.5 + glutathione 10-20 mmol / L, ulcerative colitis colon pH 4.5-5.0 + cysteine protease ≥5 ng / mL), avoiding the problem of single stimulus (such as only pH response) being easily disturbed by the normal tissue microenvironment in the body, and realizing the release mode of "precise triggering at target site, stable dormancy at non-target site".

[0114] "Pulse-slow release" dual-mode release optimizes treatment effect: Unlike the defects of traditional systems "burst release without sustained effect" or "slow release but delayed onset", the scheme realizes "pulse-slow release" dual-mode release under dual stimuli: 30%-50% of the drug is released within 1h in the pulse phase (such as 42% in Example 1, 38% in Example 2, and 35% in Example 3), rapidly increasing the drug concentration at the target site to reach the treatment threshold; 40%-60% of the drug is released within 23h in the slow release phase, maintaining the drug concentration at the target site within the effective treatment window, avoiding the toxic side effects or insufficient efficacy caused by sudden rise and fall of drug concentration. As in Example 1, dual-mode release maintains the drug concentration at the tumor site of SK-BR-3 tumor-bearing mice at IC 50(IC50) for 24 h, significantly longer than the traditional single-pulse release system (effective concentration maintenance time ≤ 8 h).

[0115] Targeting-anti-clearance bifunctional modification improves in vivo targeting and bioavailability: precise targeting reduces non-target tissue distribution: by modifying the surface of the carrier with disease-specific targeting ligands (such as RGD peptide for breast cancer, folate for colitis, and galactose for fatty liver), the high specificity of ligand binding to target cell surface receptors (such as RGD peptide binding to integrin αvβ3 with a rate of ≥80%, folate binding to folate receptors with a constant of ≥10 9 L / mol), active targeting of drugs to target sites is achieved. In Example 1, the RGD peptide-modified system was enriched in the tumor site of SK-BR-3 tumor-bearing mice by 35% of the injection dose, and the distribution in non-target organs (heart, kidney) was ≤8%; in Example 2, the folate-modified system had a drug concentration in the colon of ulcerative colitis rats that was 8 times that of the stomach, with a targeting efficiency of 82%, effectively reducing the damage to normal tissues (such as cardiotoxicity and gastrointestinal irritation) caused by the drug.

[0116] PEGylation anti-clearance modification prolongs in vivo circulation time: PEGylation modification of the surface of the carrier (molecular weight 2000-10000 Da, modification density 0.5-2 μg / cm 2 ) forms a "steric barrier" to avoid rapid clearance of the carrier by the body's mononuclear phagocyte system (MPS), significantly prolonging the in vivo circulation time of the system (half-life ≥ 12 h), which is more than 3 times longer than that of the unmodified system (half-life ≤ 4 h). As shown in Example 3, the PEGylated system has a circulation time of 15 h in non-alcoholic fatty liver mice, providing sufficient time for the drug to be fully enriched in the liver target site, with a final liver drug enrichment of 32% of the injection dose, which is more than 1 times higher than that of the unmodified system (enrichment ≤ 15%).

[0117] Multi-disease adaptability and clinical treatment safety highlight clinical value: multi-disease subtype precise adaptation: by adjusting the type of strong force unit, the type of targeting ligand, the mechanism of stimulus response, and the combination of drugs, the system can be flexibly adapted to the treatment needs of different diseases and disease subtypes: for HER2-positive breast cancer, RGD peptide targeting + Schiff base bond-hydrogen bond mediation + pH-glutathione response is used to load trastuzumab and paclitaxel (Example 1); for ulcerative colitis, folate targeting + disulfide bond-hydrogen bond mediation + pH-enzyme response is used to load ibuprofen and TNF-α siRNA (Example 2); for non-alcoholic fatty liver, galactose targeting + Zn 2+ coordination bond-hydrogen bond mediation + temperature-pH response is used to load PPAR-γ plasmid DNA and curcumin (Example 3). Multi-disease adaptability greatly expands the clinical application scenarios of the system, reducing the cost of developing separate delivery systems for different diseases.

[0118] High biocompatibility and low toxic side effects ensure treatment safety: The carrier matrix is selected from biocompatible materials (such as PLGA, chitosan quaternary ammonium salt, PEG-PCL, mesoporous silica), which have passed the biological safety verification (such as PLGA has been approved by FDA for clinical use); the preparation process does not have toxic solvent residues (such as impurities are removed by gel filtration chromatography purification), and the precise targeted delivery of drugs reduces non-target site exposure. In Example 2, the liver and kidney function indicators (ALT, AST, BUN, Cr) of rats with ulcerative colitis for 14 days of continuous administration had no obvious abnormalities, and the colon mucosa had no irritation reaction; in Example 1, the weight change rate of SK-BR-3 tumor-bearing mice during treatment was ≤5%, which proved the good biological safety of the system and reduced the risk of clinical drug use.

[0119] Strong controllability of preparation process, with potential for industrial application: The process parameters are clear and easy to scale up: The preparation steps of this scheme (such as core-shell carrier preparation, strong force construction, surface modification) are set with precise and controllable process parameters (such as temperature 25-60℃, pH 5.0-10.0, reaction time 1-12h, centrifugal speed 8000-12000rpm), and the equipment used (such as three-necked flask, centrifuge, gel filtration chromatograph) are common equipment for pharmaceutical industrialization, without special customization needs, which is convenient for scaling up from small-scale (such as 20-50mL reaction system in the examples) to pilot and industrial production (such as 100-1000L reaction kettle).

[0120] Quality controllability meets clinical standards: Through clear quality control indicators (particle size 80-400nm, PDI≤0.25, drug encapsulation efficiency≥75%, cumulative release rate≥85% under double stimulation for 24h), combined with mature detection means such as dynamic light scattering instrument, high performance liquid chromatograph, fluorescence spectrophotometer, the whole process monitoring of product quality can be realized, ensuring the batch stability (such as the particle size coefficient of variation of 3 batches of samples in Example 1≤5%, the encapsulation efficiency coefficient of variation≤3%), which meets the requirements of Good Manufacturing Practice (GMP) for drugs, laying a foundation for subsequent clinical conversion and industrial application.

[0121] The above describes the present application and its embodiments, which are not limiting, and the embodiments shown in the drawings are only one of the embodiments of the present application, and the actual structure is not limited thereto. In summary, if a person of ordinary skill in the art is inspired thereby, without departing from the spirit of the present application, similar structural modes and embodiments can be designed without creative design, which shall fall within the protection scope of the present application.

Claims

1. A strong force-mediated intelligent drug delivery system, characterized in that: It comprises a core-shell structure carrier matrix, a drug active ingredient, a strong force unit mediating the specific binding of the two, and a surface functionalization layer; the core layer of the core-shell structure carrier matrix is a mesoporous material (pore size 2-10 nm), the shell layer is a biocompatible polymer coating (thickness 10-50 nm), the overall particle size is 80-400 nm and the dispersion coefficient PDI is ≤0.25; the strong force unit is composed of "main force + auxiliary force", the main force is selected from one of coordination bond and dynamic covalent bond, the auxiliary force is selected from at least one of hydrogen bond and π-π stacking interaction, and the main force can reversibly break / weaken in response to double physiological microenvironment stimuli, and the auxiliary force cooperatively controls the drug release kinetics; Under the stimulation of double physiological microenvironment, the main force breaks, the drug-carrier binding force decreases by ≥60%, the auxiliary force maintains the stability of the carrier structure, and the drug active ingredient realizes "pulse-slow release" dual-mode release at the target site (24h cumulative release rate: 30%-50% released within 1h in the pulse phase, 40%-60% released within 23h in the slow release phase).

2. A preparation method of the strong force-mediated intelligent drug delivery system according to claim 1, characterized in that: It comprises the following steps: (1) Core-shell carrier matrix preparation: ① Mesoporous core layer preparation: using tetraethyl orthosilicate / metal organic framework material as raw material, cetyltrimethylammonium bromide / polyetheramine as template agent, sol-gel reaction at pH 8.5-10.0 and temperature 45-60℃ for 8-12h, calcination (500-600℃, 4-6h) or solvent extraction to remove the template agent, to obtain the mesoporous core layer; ② Shell coating: dispersing the mesoporous core layer in a polymer solution (concentration 1-5mg / mL), under nitrogen protection and stirring speed 300-500rpm, adding crosslinking agent (mass ratio to polymer 1:5-1:10), reacting at 30-40℃ for 6-8h, centrifuging (8000-12000rpm, 10-15min) to collect the core-shell carrier matrix; (2) Strong force unit construction and drug loading: dissolving the drug active ingredient (concentration 0.5-2mg / mL) and the main force reagent (molar ratio to drug 1:1-1:3) in buffer (pH 5.5-7.5), adding the core-shell carrier matrix (mass ratio of drug to carrier 1:5-1:20), reacting at 25-35℃ for 4-6h, then adding the auxiliary force reagent (concentration 0.1-1mg / mL), continuing to react for 2-3h, to form a drug-carrier-strong force complex; (3) Surface functionalization modification: adding targeting ligand (1:10-1:20 with carrier mass ratio) and PEGylation reagent (molecular weight 2000-10000 Da, modification density 0.5-2 μg / cm 2 ) to the above complex, reacting at 37°C for 3-5 h to achieve "targeting-anti-clearance" bifunctional modification; (4) Purification and quality control: purifying by gel filtration chromatography (eluent PBS, flow rate 1-2mL / min), collecting the target component, determining the particle size (80-400nm), PDI (≤0.25), drug encapsulation efficiency (≥70%) and stimulus-responsive release efficiency (24h cumulative release rate under double stimuli ≥85%), to obtain the intelligent drug delivery system.

3. The smart drug delivery system of claim 1, wherein: the mesoporous core layer of the core-shell structure carrier matrix is selected from one of mesoporous silica, mesoporous hydroxyapatite, metal organic framework (MOFs, such as ZIF-8, UiO-66); the shell polymer is selected from at least one of polylactic-co-glycolic acid (PLGA, lactic acid / glycolic acid molar ratio 50:50-75:25), polyethylene glycol-poly caprolactone block copolymer (PEG-PCL, PEG content 10%-30%), chitosan quaternary ammonium salt (substitution degree > 60%), and the shell polymer is grafted with active groups (amino, carboxyl or sulfhydryl) capable of reacting with the main force reagent.

4. The smart drug delivery system of claim 1, wherein: in the strong force unit: The main force is coordination bond, which is formed by metal ions (Fe 3+ , Zn 2+ or Cu 2+ with concentration of 0.1-1mmol / L) and coordination groups (hydroxyl, amino, carboxyl or phenanthroline group) in carrier shell layer / active ingredients, and the stability constant Kf is 10 8 -10 12 L / mol, and Kf drops to 10 3 -10 5 L / mol in pH 4.0-5.0 acid environment. when the main force is a dynamic covalent bond, it is selected from a Schiff base bond (formed by reaction of an aldehyde-modified carrier with an amino-modified drug, which is broken at pH 4.0-5.0 or glutathione concentration > 10 mmol / L), a disulfide bond (formed by reaction of a sulfhydryl-modified drug with a dithio succinimidyl ester-modified carrier, which is broken at glutathione concentration > 5 mmol / L or in the presence of matrix metalloproteinase-2 / 9); when the auxiliary force is a hydrogen bond, it is formed by hydroxyl / amino groups on the carrier shell and carbonyl / hydroxyl groups on the drug, with a bond energy of 2-8 kJ / mol; when it is a π-π stacking interaction, it is formed by aromatic rings (such as benzene rings, naphthalene rings) on the carrier surface and aromatic rings on the drug, with an interaction energy of 1-5 kJ / mol.

5. The smart drug delivery system of claim 1, wherein: the dual physiological microenvironment stimuli are selected from one of the following combinations: ① pH 4.5-5.5 acidic environment + glutathione concentration 10-20 mmol / L; ② temperature 39-42°C + matrix metalloproteinase-2 / 9 (concentration > 10 ng / mL); ③ pH 4.0-5.0 acidic environment + cysteine protease (concentration > 5 ng / mL); and the trigger delay time of the stimuli response is < 30 min, and the response completion time is < 2 h.

6. The smart drug delivery system of claim 1, wherein: The surface functionalization layer comprises at least one of a targeting ligand selected from folate (modification density 20-40 per μm 2 , a binding constant for folate receptor positive cells ≥10 9 L / mol), RGD peptide (sequence Arg-Gly-Asp, modification density 15-35 per μm 2 , a binding rate for integrin αvβ3 ≥80%), transferrin (modification density 5-15 per μm 2 , a targeting efficiency for transferrin receptor ≥75%), and an anti-clearance modification being PEGylation (molecular weight 5000-8000 Da, modification density 1-1.5 μg / cm 2 ), and the PEG end can be grafted with a fluorescent probe (such as FITC, Cy5) for in vivo tracking.

7. The smart drug delivery system of claim 1, wherein: The drug active ingredient is selected from one of the following combinations: ① small molecule chemotherapeutic drugs (doxorubicin, paclitaxel or cisplatin, loading amount 8%-15% mass fraction) + photosensitizer (hematoporphyrin monomethyl ether, loading amount 2%-5% mass fraction); ② antibody drugs (trastuzumab, bevacizumab, loading amount 3%-8% mass fraction) + small molecule anti-inflammatory drugs (ibuprofen, loading amount 5%-10% mass fraction); ③ nucleic acid drugs (siRNA, miRNA or plasmid DNA, loading amount 1%-3% mass fraction) + small molecule chemotherapeutic drugs (cisplatin, loading amount 5%-12% mass fraction); and the dispersion uniformity of the drug active ingredient in the delivery system is ≥90%, and there is no obvious agglomeration (agglomerated particles account for ≤5%).

8. The preparation method of the strong force mediated intelligent drug delivery system according to claim 2, characterized in that: In step (1), if the mesoporous core layer is metal organic framework material ZIF-8, the preparation process is as follows: zinc nitrate (concentration 0.1-0.5 mol / L) and 2-methyl imidazole (concentration 0.4-2 mol / L) are mixed at a molar ratio of 1:4-1:8, stirred at 25-30°C in methanol solvent for 2-4 h, centrifuged (10000 rpm, 10 min) and washed with methanol for 3-5 times, and vacuum dried (60°C, 12 h) to obtain ZIF-8 mesoporous core layer; if the shell layer is PLGA, the polymer solution is prepared by using dichloromethane / acetone mixed solvent (volume ratio 3:1-5:1), and the crosslinking agent is a mixture of 1-ethyl-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC) and N-hydroxysuccinimide (NHS) (molar ratio 1:1).

9. The preparation method of the strong force mediated intelligent drug delivery system according to claim 2, characterized in that: In step (2), if the main force is Schiff base bond, the drug active ingredient is aminylated doxorubicin (prepared by reacting doxorubicin with ethylenediamine at a molar ratio of 1:2-1:5), and the carrier shell layer is aldehyde PLGA (prepared by reacting PLGA with glutaraldehyde at a mass ratio of 10:1-20:1); the reaction is carried out at pH 6.8-7.2 and temperature 32-35°C for 5-6 h, and then an auxiliary force reagent (such as o-dihydroxybenzene, concentration 0.5 mg / mL) is added and reacted for 2.5 h to form a Schiff base bond-hydrogen bond synergistically mediated complex; in step (3), the PEGylation reagent is selected to be methoxyl polyethylene glycol-succinimidyl carbonate (mPEG-SC), and the reaction with the carrier is carried out in PBS buffer at pH 7.2-7.4; after the reaction is completed, the particle size change is monitored in real time by dynamic light scattering instrument to ensure that the particle size increase after PEGylation modification is ≤20 nm.

10. The strong force mediated intelligent drug delivery system according to claim 1, characterized in that: The application in the preparation of drugs for treating specific subtype diseases, specifically: ① for HER2 positive breast cancer, RGD peptide-PEG double modification, Schiff base bond-π-π stacking mediated delivery system is selected, loading trastuzumab and paclitaxel, through pH5.0-5.5+ glutathione 15mmol / L double stimulus response release; ② for ulcerative colitis, folic acid-PEG double modification, disulfide bond-hydrogen bond mediated delivery system is selected, loading ibuprofen and siRNA (targeting TNF-α gene), through pH4.5-5.0+ cysteine protease double stimulus response release; ③ for non-alcoholic fatty liver, galactose-PEG double modification, coordination bond (Zn 2+ )-hydrogen bond mediated delivery system is selected, loading plasmid DNA (targeting PPAR-γ gene) and curcumin, through temperature 40-42℃+pH5.0-5.5 double stimulus response release; and the enrichment amount of the delivery system in the target organ is ≥30% of the injection dose, and the distribution amount in the non-target organ is ≤10% of the injection dose.

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