Nanometer delivery system modified by antihistamine component as well as preparation method and application of nanometer delivery system

Abstract

The invention discloses an antihistamine component modified nano delivery system as well as a preparation method and application thereof. The nano delivery system comprises a nano capsule modified by an antihistamine component. The nanocapsule comprises nanoparticles formed by an acrylic acid derivative polymeric monomer, a cross-linking agent and an initiator. Specific binding of the antihistamine component and a histamine receptor in a brain area is utilized, and specific receptor mediated cross-blood brain barrier drug delivery is achieved.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of biological medicine, and particularly relates to an anti-histamine component modified nano delivery system and a preparation method and application thereof. BACKGROUND

[0002] The blood-brain barrier is composed of tightly connected brain endothelial cells, pericytes, astrocytes and neuronal cells. The blood-brain barrier precisely controls the brain microenvironment and neural activity by regulating the entry and exit of substances into the brain, and blocks almost 100% of macromolecular drugs and more than 98% of small molecular drugs from entering the brain lesion area, which brings great difficulties to the treatment of brain diseases.

[0003] In recent years, a lot of work has been done around the development of efficient brain-targeting drug delivery systems, such as adsorption-mediated transcytosis strategy, which is achieved by electrostatic interaction between positively charged drug-loaded microparticles and negatively charged blood-brain barrier biomembrane, and relies on clathrin-mediated endocytosis to transport from blood to brain unidirectionally. However, when the surface of the drug-loaded microparticles is positively charged, it may cause damage to the biomembrane of normal cells. In addition, this electrostatic interaction is non-specific, and the drug-loaded microparticles are randomly distributed in the body and are easily captured by the endothelial system in other areas, resulting in a decrease in the accumulation of drugs in the brain. The blood-brain barrier can also be temporarily opened by ultrasound to increase its permeability to drug-loaded particles, but it is highly dependent on the accuracy of equipment parameters and the proficiency of personnel. At present, although there are several physical or chemical methods to promote drug-loaded microparticles to enter the blood-brain barrier, these methods all have considerable risk of damage to the blood-brain barrier.

[0004] Histamine is an active amine substance formed by decarboxylation of histidine in the body. Mast cells, basophils and histaminergic neurons are the main places for endogenous histamine synthesis. Both endogenous and exogenous histamine can regulate various physiological processes, including contraction of intestinal smooth muscle, secretion of gastric acid, vasodilation and regulation of immune response. In the brain tissue, histamine, as a neurotransmitter, mainly comes from histaminergic neurons, which originate from the dorsal and ventral tuberomammillary nucleus of the hypothalamus, and their nerve fibers are widely distributed in the whole brain, cerebellum, posterior pituitary and spinal cord, forming a central histaminergic system to regulate the activity of central neurons and various brain functions, including pain, sleep and wakefulness, biological rhythm and cognitive function regulation. Four kinds of histamine receptors have been found: H1R, H2R, H3R and H4R, which have different distributions and physiological functions. Histamine binds to different receptors, activates G protein to convert extracellular signals to intracellular second signal system, triggers calcium release, and activates protein kinase cascade, and plays a potential role in the transmission of nociception and neurogenic inflammation. Histamine and histamine receptors can help nanoparticles enter the blood-brain barrier through the corresponding receptor highly expressed in the brain via a transporter-mediated pathway. SUMMARY

[0005] To overcome the deficiencies in the prior art, the present application provides an antihistamine component modified nano delivery system for delivering drugs to the brain, and a preparation method and application thereof.

[0006] In a first aspect of the present application, a nano delivery system is provided, which comprises an antihistamine component modified nano capsule, wherein the antihistamine component is an antihistamine drug with a carboxyl structure in the molecule.

[0007] Preferably, the modification is a covalent modification.

[0008] Preferably, the nano capsule comprises a nano particle formed by an acrylic derivative polymerization monomer, a crosslinking agent and an initiator.

[0009] Preferably, the acrylic derivative polymerization monomer is composed of one or more of acrylate or acrylamide monomers, and further preferably, the acrylic derivative polymerization monomer comprises one or more of acrylic acid salt, acrylamide, methacryloyloxy ethyl trimethyl ammonium chloride, poly(ethylene glycol) methyl ether acrylate, 2-methacryloyloxy ethyl phosphoryl choline (MPC), N-(3-aminopropyl) methacrylamide hydrochloride (APM), vinyl pyrrolidone, acryloyloxy ethyl trimethyl ammonium chloride, [2-(methacryloyloxy) ethyl] dimethyl-(3-sulfopropyl) ammonium hydroxide, 2-carboxyethyl acrylate, 3-[[2-(methacryloyloxy) ethyl] dimethylammonium] propionate, and 3-[(3-acrylamidopropyl) dimethylammonium] propionate.

[0010] More preferably, the acrylic derivative polymerization monomer comprises a combination of 2-methacryloyloxy ethyl phosphoryl choline and N-(3-aminopropyl) methacrylamide hydrochloride.

[0011] Preferably, the crosslinking agent is a biodegradable crosslinking agent, and further preferably, the crosslinking agent is selected from N,N'-methylene bis-acrylamide (BIS), ethylene glycol dimethacrylate, triethylene glycol dimethacrylate, or N,N'-bis(acryloyl) cystamine, poly(lactic acid)-poly(ethylene glycol)-poly(lactic acid) co-block diacrylate.

[0012] Preferably, the initiator includes, but is not limited to, one or more of ammonium persulfate, alkali metal persulfate, N,N,N',N'-tetramethylethylenediamine, dimethylaminopropionitrile, ferrous salt, benzoyl peroxide, 2,2'-azobisisobutyronitrile, sodium metabisulfite, azobisisobutyronitrile, cerium ammonium nitrate, benzpinacol, benzyl peroxide, or 1,2'-azobis(2,4-dimethylvaleronitrile).

[0013] In one embodiment, the initiator is ammonium persulphate (APS) and N,N,N',N'-tetramethylethylenediamine (TEMED).

[0014] Preferably, the antihistamine component includes, but is not limited to, levocetirizine, fexofenadine, bepotastine, or other components having histamine antagonistic effect and containing carboxyl structure.

[0015] Preferably, the nanocapsule has a particle size ranging from 10 to 40 (e.g., 10, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 30, 35, 40) nm.

[0016] Preferably, the molar ratio of 2-methacryloyloxyethyl phosphorylcholine, N-(3-aminopropyl) methacrylamide hydrochloride, and crosslinking agent is (5000-7000):(150-300):(500-700), which can be any of the above ratios, such as (5000, 5500, 6000, 6500, 6600, 6643, 6700, 7000):(150, 200, 250, 260, 265.75, 270, 300):(500, 550, 600, 650, 660, 664.3, 670, 700).

[0017] In a second aspect, the present application provides a method for preparing the nanodelivery system described above, which comprises:

[0018] (1) mixing an acrylic derivative polymer monomer and a crosslinking agent, and constructing a nanocapsule via free radical polymerization under the initiation of an initiator;

[0019] (2) selecting an antihistamine component containing a carboxyl structure;

[0020] (3) coupling the antihistamine component selected in step (2) with the nanocapsule of step (1) to obtain a nanodelivery system modified with the antihistamine component on the surface.

[0021] Preferably, the antihistamine component selected in step (2) is directly condensed and coupled with the nanocapsule of step (1).

[0022] Preferably, the acrylic derivative polymerization monomer is a combination of 2-methacryloyloxyethylphosphocholine and N-(3-aminopropyl) methacrylamide hydrochloride.

[0023] Preferably, the molar ratio of 2-methacryloyloxyethylphosphocholine, N-(3-aminopropyl) methacrylamide hydrochloride and crosslinking agent is (5000-7000):(150-300):(500-700), which can be any of the above ratios, for example (5000, 5500, 6000, 6500, 6600, 6643, 6700, 7000):(150, 200, 250, 260, 265.75, 270, 300):(500, 550, 600, 650, 660, 664.3, 670, 700).

[0024] Preferably, in step (3), the anti-histamine component is coupled with the nanocapsule by adding a coupling agent to synthesize N-hydroxysuccinimide active ester, and further preferably, the coupling agent includes but is not limited to 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDCI) and N-hydroxysuccinimide (NHS).

[0025] Preferably, the molar ratio of the nanocapsule to N-hydroxysuccinimide active ester is 1:(20-50), for example 1:(20, 25, 30, 35, 40, 45, 50).

[0026] In one specific embodiment, the molar ratio of 2-methacryloyloxyethylphosphocholine, N-(3-aminopropyl) methacrylamide hydrochloride and N,N'-methylenebisacrylamide is 6643:265.75:664.3, which are sequentially added to a centrifuge tube, and then N,N,N,N-tetramethyl ethylenediamine and ammonium persulfate are added, and the reaction is carried out on a rotary shaker overnight to obtain nanocapsules.

[0027] In one specific embodiment, in step (3), the anti-histamine drug with carboxyl structure is dissolved in N,N-dimethylformamide (N,N-Dimethylformamide, DMF), and an equal amount of N-hydroxysuccinimide and an equal amount of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1-(3-dimethylaminopropyl)-3-ethylcarbodiimide, EDCI) are added, and the nanocapsule is mixed with the corresponding succinimide active ester at a molar ratio of 1:40.

[0028] In a third aspect, the present application provides a use of any of the above-mentioned nano-delivery system in the preparation of a pharmaceutical composition for treating and / or preventing brain diseases.

[0029] Preferably, the pharmaceutical composition comprises the above-mentioned nano-delivery system and a drug for treating and / or preventing brain diseases, wherein the nano-delivery system delivers the drug for treating and / or preventing brain diseases.

[0030] Preferably, the drug can be a biological macromolecular drug such as a protein, a nucleic acid or a polysaccharide.

[0031] Preferably, the protein comprises a monoclonal antibody, an enzyme, an albumin, an immunoglobulin, a casein, a lipoprotein, a hemoglobin, a lysozyme, an alpha-2-macroglobulin, a fibronectin, a vitronectin, a fibrinogen, an antibody protein or a lipase.

[0032] In a specific embodiment, the albumin is serum albumin, and more preferably, the serum albumin is bovine serum albumin.

[0033] In a specific embodiment, the protein comprises a modification, and preferably, the modification reagent comprises but is not limited to 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDCI), N-hydroxysuccinimide (NHS) or N-acryloyloxysuccinimide (2-Propenoic acid, 2,5-dioxo-1-pyrrolidinyl ester, NAS).

[0034] Preferably, the molar ratio of the protein to the modification reagent is 1:(20-40), such as 1:(20, 25, 30, 35, 40).

[0035] Preferably, the pharmaceutical composition further comprises a pharmaceutically acceptable excipient and adjuvant.

[0036] Preferably, the pharmaceutical composition can be a vaccine.

[0037] In a fourth aspect, the present application provides a pharmaceutical composition comprising the above-mentioned nano-delivery system and a drug for treating and / or preventing brain diseases.

[0038] Preferably, the nano-delivery system delivers the drug for treating and / or preventing brain diseases.

[0039] Preferably, the drug can be a biological macromolecular drug such as a protein, a nucleic acid or a polysaccharide.

[0040] Preferably, the protein includes monoclonal antibody, enzyme, albumin, immunoglobulin, casein, lipoprotein, hemoglobin, lysozyme, α-2-macroglobulin, fibronectin, vitronectin, fibrinogen, antibody protein or lipase.

[0041] In one embodiment, the albumin is serum albumin, and more preferably, the serum albumin is bovine serum albumin.

[0042] In one embodiment, the protein includes a modification, and preferably, the modification reagent includes but is not limited to 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDCI), N-hydroxysuccinimide (NHS) or N-acryloyloxysuccinimide (2-Propenoic acid, 2,5-dioxo-1-pyrrolidinyl ester, NAS).

[0043] Preferably, the molar ratio of the protein to the modification reagent is 1:(20-40), such as 1:(20, 25, 30, 35, 40).

[0044] Preferably, the pharmaceutical composition further includes pharmaceutically acceptable excipients and adjuvants.

[0045] More preferably, the dosage form of the pharmaceutical composition is any one or several of liquid preparation, powder and injection, tablet; further preferably, the pharmaceutical composition is subcutaneous injection, intramuscular injection or microneedle injection.

[0046] Preferably, the pharmaceutical composition can be a vaccine.

[0047] In the fifth aspect of the present application, a preparation method of the above-mentioned pharmaceutical composition is provided, and the preparation method includes:

[0048] (1) mixing a drug, an acrylic acid derivative polymerization monomer and a crosslinking agent, and constructing a drug nanocapsule via free radical polymerization under the initiation of an initiator;

[0049] (2) selecting an antihistamine component with a carboxyl structure;

[0050] (3) coupling the antihistamine component selected in step (2) with the drug nanocapsule in step (1) to obtain a pharmaceutical composition modified with the antihistamine component on the surface.

[0051] Preferably, the drug can be a biological macromolecular drug such as protein, nucleic acid or polysaccharide.

[0052] Preferably, the drug is a protein, such as monoclonal antibody or enzyme.

[0053] More preferably, the protein is modified before step (1) starts, and further preferably, the modifying agent includes but is not limited to 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDCI), N-hydroxysuccinimide (NHS) or N-acryloyloxysuccinimide (2-Propenoic acid, 2,5-dioxo-1-pyrrolidinyl ester, NAS).

[0054] Preferably, the molar ratio of the protein to the modifying agent is 1:(20-40), such as 1:(20, 25, 30, 35, 40).

[0055] Preferably, the acrylic acid derivative polymerization monomer is a combination of 2-methacryloyloxyethyl phosphorylcholine and N-(3-aminopropyl) methacrylamide hydrochloride.

[0056] Preferably, the molar ratio of the modified protein, 2-methacryloyloxyethyl phosphorylcholine, N-(3-aminopropyl) methacrylamide hydrochloride and the crosslinking agent is 1:(5000-7000):(150-300):(500-700), which can be any of the above ratios, such as 1:(5000, 5500, 6000, 6500, 6600, 6643, 6700, 7000):(150, 200, 250, 260, 265.75, 270, 300):(500, 550, 600, 650, 660, 664.3, 670, 700).

[0057] Preferably, in step (3), the antihistamine component is synthesized by adding a coupling agent to form an N-hydroxysuccinimide active ester and then coupled with the nanocapsule, and further preferably, the coupling agent includes but is not limited to 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDCI) and N-hydroxysuccinimide (NHS).

[0058] Preferably, the molar ratio of the nanocapsule to the N-hydroxysuccinimide active ester is 1:(20-50), such as 1:(20, 25, 30, 35, 40, 45, 50).

[0059] In one specific embodiment, the molar ratio of the protein, 2-methacryloyloxyethyl phosphorylcholine, N-(3-aminopropyl) methacrylamide hydrochloride and N,N'-methylenebisacrylamide is 1:6643:265.75:664.3, which are sequentially added to a centrifuge tube, and then N,N,N,N-tetramethylethylenediamine and ammonium persulfate are added thereto, and the reaction is carried out on a rotary shaker overnight, and then the unreacted residual protein is removed by CL-4B type hydrophobic column purification to obtain the drug nanocapsule.

[0060] In one embodiment, the anti-histamine drug with carboxyl structure is dissolved in N,N- dimethylformamide (DMF) in step (3), and an equal amount of N-hydroxysuccinimide and an equal amount of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDCI) are added to mix the drug nanocapsule and the corresponding succinimide active ester at a molar ratio of 1:40.

[0061] Preferably, the concentration of the drug nanocapsule directly corresponds to the concentration of the protein inside.

[0062] The protein nanocapsule is one of the drug nanocapsules.

[0063] Preferably, the protein includes a monoclonal antibody, an enzyme, an albumin, an immunoglobulin, a casein, a lipoprotein, a hemoglobin, a lysozyme, an α-2-macroglobulin, a fibronectin, a vitronectin, a fibrinogen, an antibody protein, or a lipase.

[0064] In one embodiment, the albumin is serum albumin, and more preferably, the serum albumin is bovine serum albumin.

[0065] In one embodiment, the protein includes a modification, and preferably, the modification reagent includes but is not limited to 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDCI), N-hydroxysuccinimide (NHS), or N-acryloyloxy succinimide (2-Propenoic acid, 2,5-dioxo-1-pyrrolidinyl ester, NAS).

[0066] Preferably, the molar ratio of the protein to the modification reagent is 1:(20-40), such as 1:(20, 25, 30, 35, 40).

[0067] In a sixth aspect, the present application provides a method for treating and / or preventing a brain disease, the treatment method comprising applying the above-mentioned drug composition to a subject.

[0068] Preferably, the subject can be a human or a non-human animal.

[0069] Further, the non-human animal can be a non-human mammal.

[0070] The non-human mammal can be any one of, but not limited to, a mouse, a rat, a guinea pig, a hamster, a pig, a dog, a sheep, a monkey, a rabbit, a cat, a cow, and a horse.

[0071] The administration described herein includes, but is not limited to, intramuscular injection, subcutaneous injection, intradermal injection, intravenous injection, intra-arterial injection, intratumoral injection, intrathecal injection, intraperitoneal injection, microneedle injection, mucosal administration, oral administration, oral-nasal spray or aerosol inhalation.

[0072] The "brain disease" described in the present application includes, but is not limited to, cerebrovascular disease, inflammatory disease, intracranial space-occupying lesion, hydrocephalus, brain parasitic disease, extrapyramidal system disease, epilepsy, craniocerebral trauma, brain tumor, genetic disease or toxic and metabolic encephalopathy, etc.

[0073] Further preferably, the cerebrovascular disease includes, but is not limited to, cerebral arteriosclerosis, cerebral infarction, cerebral hemorrhage and subarachnoid hemorrhage; the inflammatory disease includes, but is not limited to, encephalitis, meningitis or meningoencephalitis; the brain parasitic disease includes, but is not limited to, cerebral schistosomiasis, cerebral echinococcosis, cerebral cysticercosis; the extrapyramidal system disease includes, but is not limited to, Parkinson's syndrome, chorea, athetosis, torsion spasm; the craniocerebral trauma includes, but is not limited to, concussion, contusion and laceration of brain; the brain tumor includes, but is not limited to, meningioma, glioma, pituitary tumor; the genetic disease includes, but is not limited to, cerebral leukodystrophy, mitochondrial encephalomyopathy; the brain degenerative disease includes, but is not limited to, Alzheimer's disease, Parkinson's disease; the toxic and metabolic encephalopathy includes, but is not limited to, ischemic-hypoxic encephalopathy, pulmonary encephalopathy, hepatic encephalopathy, renal encephalopathy, alcoholic encephalopathy.

[0074] The "treatment" described in the present application means slowing down, interrupting, stopping, controlling, reducing, or reversing the progression or severity of a sign, symptom, disorder, condition, or disease after the disease has begun to develop, but does not necessarily involve the complete elimination of all disease-related signs, symptoms, conditions, or disorders.

[0075] The "effective amount" described in the present application refers to the amount or dose of the drug of the present application that provides the desired treatment or prevention after being administered to an individual or an organ in a single or multiple doses.

[0076] The term "comprising" or "including" in the present application is an open description containing the specified components or steps, and other specified components or steps that do not materially affect.

[0077] The "individual" described in the present application can be a human or a non-human animal, and the non-human animal can be a mouse, a cow, a sheep, a rabbit, a pig, a monkey, and the like non-human mammal.

[0078] The beneficial effects of the present application:

[0079] The application provides a nano delivery system for brain drug delivery, which can be used for treating brain diseases by biological macromolecular drugs such as proteins, nucleic acids and polysaccharides. It is found through research that the nano delivery system can effectively transport drugs such as model macromolecules (bovine serum albumin) to the brain region. Compared with ordinary bovine serum albumin without being encapsulated and modified by capsules, the enrichment amount of BSA in the brain is obviously increased after being encapsulated into nano capsules modified by antihistamine drugs.

[0080] Meanwhile, the application is based on the high affinity of antihistamine drugs to the highly expressed histamine receptors on the central nervous system, and the combination of the antihistamine drugs with nano capsules capable of improving the stability of the drugs forms a new drug carrier for efficiently crossing the blood-brain barrier mediated by histamine receptors. The delivery strategy provides a new solution and selection for brain disease treatment. BRIEF DESCRIPTION OF DRAWINGS

[0081] Figure 1 : Levocetirizine modified protein nano capsule steps.

[0082] Figure 2 : Fexofenadine modified protein nano capsule steps.

[0083] Figure 3 : Betahistine modified protein nano capsule steps.

[0084] Figure 4 : DLS particle size determination results of BSA nano capsules and BSA.

[0085] Figure 5 : Zeta potential determination results of BSA nano capsules and BSA.

[0086] Figure 6 : Fluorescence imaging diagram of a mouse administered through a tail vein. DETAILED DESCRIPTION

[0087] In order for those skilled in the art to better understand the technical solutions in the application, the technical solutions in the embodiments of the application will be clearly and completely described below. Obviously, the described embodiments are only some of the embodiments of the application, but not all the embodiments. Based on the embodiments in the application, all other embodiments obtained by those skilled in the art without creative labor should belong to the protection scope of the application.

[0088] The materials, reagents and instruments used in the following embodiments can be obtained from commercial channels if not otherwise specified. Embodiment 1: Synthesis of protein nano capsules

[0089] In this embodiment, bovine serum albumin is used as a model protein to prepare drug nano capsules as an example of active drugs: 1. Synthesis of protein nano capsules

[0090] Preparation of bovine serum albumin nanocapsules (nBSA):

[0091] Weigh 40 mg of BSA and dissolve it in 2 mL of 1×PBS solution at pH 7.4 to prepare a 20 mg / mL BSA solution. Place the solution in a dialysis bag with a molecular weight cutoff of 10 kDa and dialyze it repeatedly in 1500 mL of 1×PBS solution at pH 7.4. Measure the absorbance of the BSA solution at 280 nm and calculate the protein concentration using the Lambert-Beer law. Prepare a 10 mg / mL N-acryloyloxysuccinimide (NAS) solution in dimethyl sulfoxide (DMSO). Slowly add the NAS solution at a BSA:NAS molar ratio of 1:30, stir thoroughly, and react at 4°C in the dark for 4 hours. Place the solution in a dialysis bag with a molecular weight cutoff of 10 kDa and dialyze it overnight in 1×PBS solution at pH 7.4 to remove unreacted NSA. Collect the double-bonded BSA, denoted as BSA-NAS.

[0092] The concentration of BSA-NAS was determined using the BCA (Bicinchonininc Acid) method. First, a 5 mg / mL BSA solution was prepared, followed by serial dilutions. After adding the prepared BCA reagent, the solution was incubated in a metal bath at 37°C for 30 min, and then immediately placed in an ice bath to terminate the reaction. The absorbance at 562 nm was measured, and a standard curve of concentration versus absorbance was plotted, yielding the linear regression equation y = kx + b. Similarly, BSA-NAS was diluted to a fixed factor, incubated in a metal bath at 37°C for 30 min, and then immediately placed in an ice bath to terminate the reaction. The absorbance at 562 nm was measured, and the measured absorbance of BSA-NAS was substituted into the linear regression equation to calculate x. Multiplying x by the dilution factor gives the concentration of BSA-NAS.

[0093] BSA-NAS was mixed with MPC, APM and BIS in a molar ratio of 1:6643:265.75:664.3, and the volume was adjusted to 1 mL with 1×PBS at pH=7.4. Then, TEMED and APS were added sequentially to initiate in situ free radical polymerization. The reaction was carried out at 4°C for 4 h to obtain unpurified nBSA.

[0094] Unpurified nBSA was placed in a dialysis bag with a molecular weight cutoff of 30 kDa and repeatedly dialyzed in 1×PBS solution at pH 7.4 to remove unreacted monomers, cross-linking agents, and initiators. Unencapsulated BSA was removed using a phenyl-agarose CL-4B hydrophilic-hydrophobic protein column. Successfully encapsulated nanocapsules were collected and concentrated using Millipore ultrafiltration centrifuge tubes to obtain purified nBSA.

[0095] 2. Characterization of nBSA

[0096] Particle size testing:

[0097] BSA and nBSA were diluted to 0.4 mg / mL with 1xPBS solution of pH = 7.4, 1 mL was added to disposable sample cell, parameters were adjusted, and the particle size was tested by nanoparticle size and zeta potential analyzer, and the test temperature was 25°C.

[0098] Zeta potential test:

[0099] BSA and nBSA were diluted to 0.4 mg / mL with 1xPBS solution of pH = 7.4, 800 μL was added to a Malvern potential cell, parameters were adjusted, and the surface potential was tested by nanoparticle size and zeta potential analyzer, and the test temperature was 25°C.

[0100] The particle size results of dynamic light scattering are shown in Figure 4 , and the results show that the particle size of nBSA is about 20 nm, which is significantly larger than that of natural BSA (<10 nm), and the change of zeta potential on the surface of the protein microparticle is shown in Figure 5 , which jointly proves the successful synthesis of nBSA.

[0101] Preparation and application of pharmaceutical composition

[0102] Coupling of antihistamine component and protein nanocapsule:

[0103] 1 equivalent of antihistamine component, 1 equivalent of N-hydroxysuccinimide, and 1 equivalent of EDCI were added to a certain amount of DMF, and stirred in the dark for 6 hours. According to the ratio of 40:1 of succinimidyl active ester to protein nanocapsule in Example 1, they were added, and reacted in the dark at 4°C for 4 hours. Desalting purification was performed using an ultrafiltration centrifuge tube with a molecular weight cutoff of 50 KDa. The antihistamine component modified protein nanocapsule was obtained. Exemplarily, levocetirizine, fexofenadine, and bepotastine were selected as antihistamine components, and the processes of obtaining protein nanosystems (one kind of pharmaceutical composition) are shown in Figures 1-3 .

[0104] Cy5.5 labeling

[0105] Under light shielding conditions, Cy5.5 succinimidyl ester FMSO solution was added according to the ratio of 1:3 of protein and fluorescent dye molecule, and after shaking reaction for 2 hours, purification was performed by using an ultrafiltration centrifuge tube with a molecular weight cutoff of 50 KDa. After ultrafiltration purification, the container of the protein nanocapsule was wrapped with tin paper, and stored at -20°C for later use.

[0106] Verification of brain targeting effect of nanodelivery system

[0107] The Cy5.5-labeled drug composition was administered to mice via tail vein injection to verify whether the nanodelivery system has a brain-targeting effect. After administration, in vivo imaging of mice was performed at 30 min, 1 h, 2 h, 4 h, 6 h, 8 h, 16 h, and 24 h, and the targeting effect was verified by the fluorescence effect in the brain.

[0108] Experimental results:

[0109] like Figure 6 As shown, in vivo imaging in mice verified that the synthesized drug composition could reach the brain 30 minutes after administration, indicating that the nanodelivery system of the present invention can achieve the effect of delivering drugs to the brain, further demonstrating the brain-targeting ability of the drug-delivering nanocapsules.

[0110] Although the present invention has been described in detail by way of preferred embodiments, the present invention is not limited thereto. Various equivalent modifications or substitutions can be made to the embodiments of the present invention by those skilled in the art without departing from the spirit and essence of the invention, and such modifications or substitutions should all be within the scope of the present invention. Any variations or substitutions that can be easily conceived by those skilled in the art within the technical scope disclosed in the present invention should also be covered within the protection scope of the present invention.

Claims

1. A nanodelivery system, characterized in that, The nano delivery system comprises a nano capsule modified by an antihistamine component, wherein the antihistamine component is an antihistamine drug derivative with a carboxyl structure in the molecule.

2. The nanodelivery system of claim 1, wherein, The nano capsule comprises nanoparticles formed by acrylic derivative polymer monomers, a crosslinking agent and an initiator.

3. The nanodelivery system according to any one of claims 1-2, wherein, The acrylic derivative polymer monomers are composed of one or more of acrylate or acrylamide monomers, preferably, the acrylic derivative polymer monomers comprise one or more of acrylic acid salt, acrylamide, methacryloyloxyethyl trimethylammonium chloride, poly(ethylene glycol) methyl ether acrylate, 2-methacryloyloxyethyl phosphorylcholine, N-(3-aminopropyl) methacrylamide hydrochloride, vinyl pyrrolidone, acryloyloxyethyl trimethylammonium chloride, [2-(methacryloyloxy)ethyl]dimethyl-(3-sulfopropyl) ammonium hydroxide, 2-carboxyethyl acrylate, 3-[[2-(methacryloyloxy)ethyl]dimethylammonium] propionate, 3-[(3-acrylamidopropyl)dimethylammonium] propionate, and further preferably, the acrylic derivative polymer monomers comprise a combination of 2-methacryloyloxyethyl phosphorylcholine and N-(3-aminopropyl) methacrylamide hydrochloride. Preferably, the crosslinking agent is a biodegradable crosslinking agent, and further preferably, the crosslinking agent is selected from N,N'-methylenebisacrylamide, ethylene glycol dimethacrylate, triethylene glycol dimethacrylate, or N,N'-bis(acryloyl)cystamine, acrylamide-polypeptide structure-acrylamide, poly(lactic acid)-poly(ethylene glycol)-poly(lactic acid) co-block diacrylate. Preferably, the initiator comprises one or more of ammonium persulfate, alkali metal persulfate, N,N,N',N'-tetramethylethylenediamine, dimethylaminopropionitrile, ferrous salt, benzoyl peroxide, 2,2'-azobisisobutyronitrile, sodium metabisulfite, azobisisobutyronitrile, cerium ammonium nitrate, benzpinacol, benzyl peroxide, or 1,2'-azobis(2,4-dimethylvaleronitrile).

4. A method of preparing the nanodelivery system according to any one of claims 1 to 3, characterized in that, The method comprises: (1) mixing the acrylic derivative polymer monomers and the crosslinking agent, and constructing the nano capsule through radical polymerization under the initiation of a chemical initiator or a physical initiation condition; (2) selecting an antihistamine component with a carboxyl structure; (3) coupling the antihistamine component selected in step (2) with the nano capsule in step (1) to obtain the nano delivery system modified by the antihistamine component on the surface.

5. The production method according to claim 4, characterized by, The acrylic derivative polymer monomers comprise a combination of 2-methacryloyloxyethyl phosphorylcholine and N-(3-aminopropyl) methacrylamide hydrochloride.

6. The production method according to claim 5, wherein The molar ratio of the 2-methacryloyloxyethyl phosphorylcholine, the N-(3-aminopropyl) methacrylamide hydrochloride and the crosslinking agent is (5000-7000):(150-300):(500-700).

7. The method of any one of claims 4-6, wherein, In the step (3), the anti-histamine component is coupled with the nanocapsule by adding a coupling agent to synthesize N-hydroxysuccinimide active ester, preferably, the coupling agent includes 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride and N-hydroxysuccinimide. Further preferably, the molar ratio of the nanocapsule to N-hydroxysuccinimide active ester is 1: (20-50).

8. Use of the nanodelivery system according to any one of claims 1-3 in the preparation of a pharmaceutical composition for treating and / or preventing brain diseases.

9. A pharmaceutical composition, characterized by, The pharmaceutical composition comprises the nanodelivery system according to any one of claims 1-3 and a drug for treating and / or preventing brain diseases; Preferably, the nanodelivery system delivers the drug for treating and / or preventing brain diseases; Preferably, the drug can be a biological macromolecular drug such as a protein, nucleic acid or polysaccharide.

10. The use of claim 8 or the pharmaceutical composition of claim 9, wherein, The brain diseases include cerebrovascular diseases, inflammatory diseases, intracranial space-occupying lesions, hydrocephalus, brain parasitic diseases, extrapyramidal diseases, epilepsy, craniocerebral trauma, brain tumors, genetic diseases or poisoning and metabolic encephalopathy.

11. A process for the preparation of a pharmaceutical composition as claimed in claim 9 or 10, characterized in that, The preparation method comprises: (1) mixing a drug, an acrylic derivative polymerization monomer and a crosslinking agent, and initiating under a chemical initiator or a physical initiation condition to construct a drug nanocapsule via free radical polymerization; (2) selecting an anti-histamine component with a carboxyl structure; (3) coupling the anti-histamine component selected in the step (2) with the drug nanocapsule in the step (1) to obtain a drug composition modified with the anti-histamine component on the surface.

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