Nano material for targeted regulation of intracellular pH value, and preparation method and application thereof

Abstract

The invention discloses a nano material for targeted regulation of intracellular pH value and a preparation method and application thereof. In order to solve the problem of cell dysfunction caused by pH value imbalance in cells, the nano vesicles based on biomimetic modification of the cell membranes are provided, pH buffer salt is entrapped by the cell membranes of target cells, and the target cells are specifically recognized. The target cells comprise related cells of a motor system, an endocrine system or a digestive system. The pH buffer salt is a phosphate compound, a carbonate compound, a citrate compound or a succinate compound. The preparation method comprises the following steps: extracting a targeted cell membrane through a differential centrifugation method, carrying out ultrasonic fusion on the targeted cell membrane and a buffer salt solution to prepare the nano material, applying the nano material to preparation of drugs for type 2 diabetes, type 2 inflammatory bowel disease or osteoporosis, specifically targeting corresponding target cells, releasing a regulator, and regulating the intracellular pH value of the target cells.

Description

Technical Field

[0001] The present invention belongs to the technical field of drug delivery and relates to the regulation of intracellular pH value, and in particular to a nanomaterial targeted for regulating intracellular pH value and a preparation method and application thereof. Background Art

[0002] The normal progress of biochemical reactions depends on a strictly regulated acid-base environment, and intracellular pH homeostasis is crucial for maintaining normal physiological functions of cells. Once the intracellular pH value fluctuates abnormally, whether it is acidification or alkalinization, it may lead to cellular metabolic disorders, abnormal signal transduction and functional disorders, thereby affecting the body's homeostasis and even inducing diseases. Diseases of the musculoskeletal system and the endocrine system are closely related to the imbalance of intracellular pH. Therefore, precise pH regulation for different cell states, that is, restoring the acid-base homeostasis of normal cells, or inducing apoptosis of diseased cells by artificially intervening in their intracellular pH, is an important strategy for treating related diseases.

[0003] At present, therapeutic strategies for intracellular pH regulation still have many limitations. Traditional acid-base regulating drugs lack targeting. After entering the body, they act on both normal cells and diseased cells. This not only affects the physiological pH balance of normal cells and causes a series of side effects, but may also reduce the therapeutic effect due to non-specific effects, making it difficult to achieve precise regulation. In addition, many drugs are easily metabolized or cleared before reaching the target cells, resulting in insufficient effective concentrations, further limiting the feasibility of treatment. Therefore, the development of a pH regulation strategy that is both highly targeted and controllable to achieve the restoration of normal cell function and the selective elimination of diseased cells has become a scientific problem that needs to be solved urgently. Summary of the Invention

[0004] In response to the shortcomings of the existing technology, the present invention proposes a nanomaterial for targeted regulation of intracellular pH value, its preparation method and application, and designs a nano cell membrane encapsulating a pH regulator. Through bionic modification, the targeted recognition ability is enhanced, so that it can accurately act on specific cell groups, achieve the effect of accurately regulating the intracellular pH value, intervene in the homeostasis of diseased cells, and reduce unnecessary interference with normal cells.

[0005] In a first aspect, the present invention provides a nanomaterial for targeted regulation of intracellular pH, wherein the nanomaterial is a cell membrane-targeted nanovesicle encapsulating a pH buffer salt.

[0006] Preferably, the pH buffer salt is a phosphate, carbonate, citrate or succinate compound.

[0007] Preferably, the pH buffer salt is at least one of disodium hydrogen phosphate, dipotassium hydrogen phosphate, diammonium hydrogen phosphate, sodium dihydrogen phosphate, potassium dihydrogen phosphate, ammonium dihydrogen phosphate, sodium bicarbonate, potassium bicarbonate, ammonium bicarbonate, sodium carbonate, potassium carbonate, ammonium carbonate, sodium citrate, potassium citrate, magnesium citrate, ferric citrate, sodium succinate, potassium succinate, and magnesium succinate.

[0008] Preferably, the target cell membrane is selected from the cell membranes of cells related to the motor system, endocrine system, and digestive system.

[0009] In a second aspect, the present invention provides a method for preparing the nanomaterial for targeted regulation of intracellular pH value, wherein the pH buffer salt is mixed with the targeted cell membrane by ultrasound, and the mixture is filtered and sterilized after low-temperature dialysis to obtain the nanomaterial for targeted regulation of intracellular pH value.

[0010] In a third aspect, the present invention provides an application of the nanomaterial for targeted regulation of intracellular pH, wherein the nanomaterial is used to specifically identify target cells, release regulators, and targetedly regulate the intracellular pH of target cells.

[0011] The present invention has the following beneficial effects:

[0012] 1) The material structure of the present invention is modular, adaptable to a variety of cell membrane sources and buffer systems, and has high expansion potential.

[0013] 2) The present invention specifically targets the pH-sensitive pathway during osteoclast precursor differentiation through a pH-responsive mechanism, achieving precise release under the intracellular acidification state of the target cells and avoiding nonspecific damage to osteoblasts and other bone cells.

[0014] 3) The present invention simultaneously blocks the H+ secretion activity of the V-type ATPase proton pump, inhibiting the formation of the acidic microenvironment of the bone resorption cavity from the source, intervening in the early critical stage of osteoclast differentiation, inhibiting bone resorption from the source, and realizing the dual-pathway regulation of "proton pump activity-intracellular pH homeostasis".

[0015] 4) In vitro and in vivo experiments have demonstrated that the present invention can significantly inhibit the number of osteoclasts and the area of ​​bone erosion, and improve the trabecular structure and bone mass in an osteoporosis model.

[0016] 5) The present invention is applicable to multiple disease models. In the type 2 diabetes model, it can target pancreatic β cells, alleviate high glucose-induced intracellular acidification, and delay functional decline. In type 2 inflammatory bowel disease, it can target inflammatory macrophages, regulate the acidic microenvironment, and intervene in the pro-inflammatory polarization process. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] Figure 1 The intracellular pH changes during the differentiation of precursor osteoclasts.

[0018] Figure 2 Extraction and characterization of precursor osteoclast membranes.

[0019] Figure 3 Characterization of Nanomaterials for Targeting Precursor Osteoclasts

[0020] Figure 4 TRAP staining for the inhibitory effect of nanomaterials targeting precursor osteoclasts on osteoclasts.

[0021] Figure 5 Scanning electron microscopy of the inhibitory effect of nanomaterials targeting precursor osteoclasts on osteoclasts.

[0022] Figure 6 Effect of nanomaterials targeting precursor osteoclasts on the intracellular pH of precursor osteoclasts.

[0023] Figure 7 Transcriptome analysis of osteoclast effects of nanomaterials targeting pre-osteoclasts.

[0024] Figure 8 Micro-CT of the inhibitory effect of precursor osteoclast-targeted nanomaterials on OVX-induced osteoporosis in mice.

[0025] Figure 9 HE staining and TRAP staining showed the inhibitory effect of nanomaterials targeting precursor osteoclasts on OVX-induced osteoporosis in mice. DETAILED DESCRIPTION

[0026] The present invention will be further explained below with reference to the accompanying drawings. It should be understood that the following examples are only used to further illustrate the present invention and cannot be understood as limiting the scope of protection of the present invention. Some non-essential improvements and adjustments made by those skilled in the art based on the above content of the present invention fall within the scope of protection of the present invention. The specific process parameters and the like in the following examples are only examples within a suitable range, that is, those skilled in the art can make appropriate selections based on the description herein, and are not necessarily limited to the specific values ​​​​in the following examples.

[0027] A nanomaterial for targeted regulation of intracellular pH value, wherein the nanomaterial is a cell membrane biomimetic nanovesicle encapsulating pH buffer salt.

[0028] The pH buffer salt is a phosphate, carbonate, citrate or succinate compound. Specifically, it can be a mixture of one or more of disodium hydrogen phosphate, dipotassium hydrogen phosphate, diammonium hydrogen phosphate, sodium dihydrogen phosphate, potassium dihydrogen phosphate, ammonium dihydrogen phosphate, sodium bicarbonate, potassium bicarbonate, ammonium bicarbonate, sodium carbonate, potassium carbonate, ammonium carbonate, sodium citrate, potassium citrate, magnesium citrate, ferric citrate, sodium succinate, potassium succinate, and magnesium succinate.

[0029] The targeted cell membrane is the cell membrane of the target cell to be delivered, such as the cell membrane of cells related to the motor system, endocrine system, and digestive system. Specifically, it can be a chondrocyte membrane, an osteoblast membrane, an osteoclast membrane, a muscle cell membrane, a mesenchymal stem cell membrane, a pancreatic β cell membrane, a pancreatic α cell membrane, an adrenal medullary cell membrane, a thyroid follicular cell membrane, a parathyroid cell membrane, a gastric epithelial cell membrane, a liver cell membrane, a pancreatic acinar cell membrane, or an intestinal epithelial cell membrane.

[0030] The method for preparing the nanomaterial targeted for regulating intracellular pH value specifically comprises the following steps:

[0031] Step 1: Extract target cell membrane by differential centrifugation

[0032] Resuspend the desired targeted cells in TM buffer pre-cooled at 4°C and use a micro-extruder to extrude the cells for 40 cycles. Mix the cell lysis solution with a 1.00 mol / L sucrose solution at a volume ratio of 3:1 to prepare a sucrose-TM buffer mixture with a final concentration of 0.25 mol / L. Centrifuge the sucrose-TM buffer mixture at 2000×g for 10 minutes and collect the supernatant. Centrifuge the supernatant at 3000×g for 30 minutes, discard the supernatant, and take the precipitate. Resuspend the precipitate with 0.25 mol / L sucrose solution and centrifuge again at 3000×g for 30 minutes. The resulting precipitate is the desired targeted cell membrane component and is stored at 4°C for later use.

[0033] The TM buffer comprises mannitol, sucrose, Tris, MgCl2, KCl, PMSF, EDTA-free protease inhibitors, DNAse and RNAse, and has a pH of 7.4. The concentration of Tris is 5 to 30 mmol / L, and the concentration of MgCl2 is 1 to 20 mmol / L.

[0034] Step 2: Preparation of cell membrane biomimetic nanovesicles loaded with pH buffer salt compounds

[0035] Add 10.00 mL of 0.5 mol / L pH buffered saline solution to the targeted cell membrane, shake and hydrate at 37°C for 30 minutes, dialyze in a dialysis bag for 48 hours, filter through a 0.22 μm sterile filter membrane to obtain the nanomaterial for targeted regulation of intracellular pH value, and store at 4°C in the dark.

[0036] Example 1

[0037] This example prepares a nanomaterial that targets and regulates intracellular pH and is injected into the tail vein as a drug for treating type 2 diabetes. The nanomaterial is a cell membrane nanovesicle loaded with sodium citrate, and the average particle size of the nanomaterial is 150 nanometers. The specific preparation steps are as follows:

[0038] Step 1: Extract pancreatic β cell membranes using differential centrifugation

[0039] Resuspend the pancreatic β cells in TM buffer pre-chilled at 4°C and extrude the cells using a microextruder for 40 cycles. Mix the cell lysis solution with 1.00 mol / L sucrose solution at a volume ratio of 3:1 to prepare a sucrose-TM buffer mixture with a final concentration of 0.25 mol / L. Centrifuge the sucrose-TM buffer mixture at 2000 × g for 10 minutes and collect the supernatant. Centrifuge the supernatant at 3000 × g for 30 minutes, discard the supernatant, and collect the pellet. Resuspend the pellet in 0.25 mol / L sucrose solution and centrifuge again at 3000 × g for 30 minutes. The resulting pellet is the pancreatic β cell membrane fraction and store at 4°C until needed.

[0040] In the TM buffer, the concentration of Tris is 10 mmol / L, and the concentration of MgCl2 is 1 mmol / L.

[0041] Step 2: Preparation of cell membrane biomimetic nanovesicles loaded with sodium citrate

[0042] Add 10.00 mL of 1 mol / L sodium citrate solution to the pancreatic β cell membrane, shake and hydrate at 37°C for 30 minutes, dialyze in a dialysis bag for 48 hours, and filter through a 0.22 μm sterile filter membrane to obtain a sodium citrate-encapsulated nanomaterial for targeted regulation of the pH value in pancreatic β cells. Store at 4°C in the dark.

[0043] The nanomaterials prepared in this example achieve precise targeting of pancreatic β cells through biomimetic modification of pancreatic β cell membranes, thereby increasing the intracellular pH of pancreatic β cells from 7.0 in the diabetic state to the normal physiological level of 7.4, making K + ATP channels are more likely to close, promoting cell depolarization and increasing Ca 2 + Influx optimizes intracellular acid-base balance, thereby enhancing K + The regulatory effect of ATP channels enhances glucose-stimulated insulin secretion. At the same time, pH restoration stabilizes the acidified state of insulin vesicles, promotes insulin storage and secretion, alleviates endoplasmic reticulum stress, and protects β-cell function. Maintaining a pH of 7.4 ensures optimal insulin secretion and avoids the negative effects of excessive alkalinization, thereby achieving pancreatic islet function repair in type 2 diabetes. Therefore, the nanomaterials targeted at regulating intracellular pH can be applied to the treatment of type 2 diabetes.

[0044] Example 2

[0045] This example prepares a nanomaterial that targets and regulates intracellular pH as a drug for treating type 2 inflammatory bowel disease. The nanomaterial is a cell membrane nanovesicle containing sodium bicarbonate, and the average particle size of the nanomaterial is 150 nanometers. The specific preparation steps are as follows:

[0046] Step 1: Extract intestinal epithelial cell membranes using differential centrifugation

[0047] Resuspend the intestinal epithelial cells in TM buffer pre-cooled at 4°C and use a micro-extruder to extrude the broken cells for 40 cycles. Mix the cell disruption solution with a 1.00 mol / L sucrose solution at a volume ratio of 3:1 to prepare a sucrose-TM buffer mixture with a final concentration of 0.25 mol / L. Centrifuge the sucrose-TM buffer mixture at 2000×g for 10 minutes and collect the supernatant. Centrifuge the supernatant at 3000×g for 30 minutes, discard the supernatant, and take the precipitate. Resuspend the precipitate with a 0.25 mol / L sucrose solution and centrifuge again at 3000×g for 30 minutes. The resulting precipitate is the intestinal epithelial cell membrane component and is stored at 4°C for later use.

[0048] Step 2: Preparation of cell membrane biomimetic nanovesicles loaded with sodium bicarbonate

[0049] 10.00 mL of 0.5 mol / L sodium bicarbonate solution was added to the intestinal epithelial cell membrane, shaken and hydrated at 37°C for 30 minutes, dialyzed in a dialysis bag for 48 hours, and filtered through a 0.22 μm sterile filter membrane to obtain a nanomaterial that targets the regulation of intracellular pH value and stored at 4°C in the dark.

[0050] The nanomaterial prepared in this embodiment is modified by biomimetic modification of the intestinal epithelial cell membrane to achieve precise targeting of intestinal epithelial cells. By administering the drug by gavage, the pH balance of intestinal epithelial cells is precisely regulated, tight junction proteins are stabilized, intestinal permeability is reduced, and the release of pro-inflammatory factors by macrophages and T cells is inhibited, thereby improving immune imbalance to repair the barrier function destroyed by acidification of epithelial cells. In addition, regulating the pH of the microenvironment promotes the growth of beneficial bacteria and reduces the metabolic toxicity of pathogens, thereby achieving precise intervention in inflammatory bowel disease, with the therapeutic advantages of high targeting and low side effects. Therefore, the nanomaterial that targets and regulates the intracellular pH value can be applied to the treatment of type 2 inflammatory bowel disease.

[0051] Example 3

[0052] This example prepares a nanomaterial that targets and regulates intracellular pH and is injected into the tail vein as a drug for treating osteoporosis. The nanomaterial is a cell membrane nanovesicle loaded with sodium hydrogen phosphate, with an average particle size of 150 nanometers. The specific preparation steps are as follows:

[0053] Step 1: Extraction of precursor osteoclast membranes by differential centrifugation

[0054] Confocal fluorescence imaging of the precursor osteoclasts during differentiation was performed, and the intracellular pH values ​​on days 0, 1, 3, and 5 were analyzed. Figure 1 As shown, the intracellular pH of osteoclast precursors first decreases and then increases during differentiation. Preosteoclast precursors were resuspended in 4°C pre-chilled TM buffer and ruptured using a microextruder with 40 cycles of extrusion. The cell lysis solution was mixed with 1.00 mol / L sucrose solution at a volume ratio of 3:1 to prepare a sucrose-TM buffer mixture with a final concentration of 0.25 mol / L. The sucrose-TM buffer mixture was centrifuged at 2000 × g for 10 minutes, and the supernatant was collected. The supernatant was centrifuged at 3000 × g for 30 minutes, and the supernatant was discarded to obtain the pellet. The pellet was resuspended in 0.25 mol / L sucrose solution and centrifuged again at 3000 × g for 30 minutes. The resulting pellet is the osteoclast precursor membrane fraction and stored at 4°C until further use.

[0055] The extracted osteoclast precursor membranes were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Figure 2 As shown, the extracted cell membrane protein only contains cell membrane protein Na + / K + -ATPase, does not contain cytoplasmic, nuclear or other organelle proteins.

[0056] Step 2: Preparation of cell membrane biomimetic nanovesicles loaded with disodium hydrogen phosphate

[0057] 10.00 mL of 0.5 mol / L disodium hydrogen phosphate solution was added to the prostate osteoclast membrane, and the mixture was shaken and hydrated at 37°C for 30 minutes. The mixture was dialyzed in a dialysis bag for 48 hours, and filtered through a 0.22 μm sterile filter membrane to obtain the nanomaterial for targeted regulation of intracellular pH value. The nanomaterial was stored at 4°C in the dark.

[0058] The particle size and potential of the precursor osteoclast membrane nanovesicles loaded with sodium hydrogen phosphate were measured using a particle size potential tester. Figure 3 As shown in (a), the particle size is 97.3nm and the potential is -25.9mV. Under cryo-transmission electron microscopy, the material is uniformly spherical, as shown in Figure 3 As shown in (b) in .

[0059] The precursor osteoclast membrane nanovesicles Na2HPO4@pOCm loaded with disodium hydrogen phosphate were added into the osteoclast induction system and compared with the osteoclast OCs induction group alone. Figure 4As shown in Figure 3, TRAP staining results showed that the number and area of ​​osteoclasts in the Na2HPO4@pOCm group were significantly inhibited, indicating that Na2HPO4@pOCm has a significant inhibitory effect on osteoclasts.

[0060] Na2HPO4@pOCm was added to the osteoclast induction system cultured in bovine bone slices, and compared with the osteoclast OCs induction group cultured in bovine bone slices. Figure 5 As shown in the figure, scanning electron microscopy results showed that the number and area of ​​bone resorption areas in the Na2HPO4@pOCm group were significantly inhibited, suggesting that Na2HPO4@pOCm has a significant inhibitory effect on osteoclasts.

[0061] Na2HPO4@pOCm was added to the osteoclast induction system and compared with the osteoclast OC system alone and the bone marrow-derived macrophage BMM system alone. Flow cytometry and confocal microscopy results showed that Na2HPO4@pOCm increased the OC cytoplasmic pH value. Figure 6 As shown. Upstream regulatory analysis of differentially expressed genes after adding OCs and Na2HPO4@pOCm showed that the predicted transcription factor with the highest Z score was forkhead box protein M1 (FOXM1), with a change of 10.09 times. qPCR results showed that the FOXM1 transcript level in the Na2HPO4@pOCm group was reduced, as shown in Figure 7 shown.

[0062] Test Example 1

[0063] This test example 1 verifies the inhibitory effect of the precursor osteoclast membrane nanovesicles Na2HPO4@pOCm loaded with disodium hydrogen phosphate prepared in Example 3 on osteoporosis induced by the ovariectomy mouse model OVX. The specific steps are as follows:

[0064] Step 1: Construction and grouping of animal disease models

[0065] Ten-week-old C57BL / 6 female mice were divided into the following four groups:

[0066] a) Sham surgery and tail vein injection of normal saline;

[0067] b) Ovariectomy and tail vein injection of normal saline;

[0068] c) Ovariectomy was performed and the tail vein was injected with sodium chloride-loaded precursor osteoclast membrane nanovesicles NaCl@pOCm;

[0069] d) Ovariectomy was performed and the cells were injected into the tail vein with precursor osteoclast membrane nanovesicles Na2HPO4@pOCm loaded with disodium hydrogen phosphate.

[0070] Step 2: Implementation

[0071] Each group of animals underwent surgery and, starting one week later, received tail vein injections of 0.025 ml / g of the drug every two days for two weeks. Four weeks after the end of drug administration, femurs and tibias were removed from each group for analysis.

[0072] Step 3: Micro-CT analysis results

[0073] The femur and tibia Micro-CT of each experimental group were compared. Figure 8 As shown in the figure, the bone mass, trabecular number and trabecular space indexes of group d) were significantly better than those of group b).

[0074] Step 4: HE staining and TRAP staining analysis results

[0075] HE staining and TRAP staining of femur and tibia of each experimental group were compared. Figure 9 As shown, group d) showed significantly better bone mass, osteoclast area, and osteoclast number than group b). Combined with the results of Micro-CT analysis, this suggests that the sodium hydrogen phosphate-loaded precursor osteoclast membrane nanovesicles Na2HPO4@pOCm prepared in Example 3 can effectively treat bone loss and osteoclast metabolism in OVX mice, thereby treating osteoporosis.

[0076] In summary, the nanomaterial prepared in Test Example 3 achieves precise targeting of precursor osteoclasts through biomimetic modification of the precursor osteoclast membrane. When osteoclast differentiation is initiated, intracellular acidification triggers membrane fusion to release Na2HPO4, neutralizes cytoplasmic H+, and destroys the lysosomal H+ gradient mediated by V-type ATPase, thereby inhibiting podosome formation and cathepsin K activation. At the same time, the reduction in H+ concentration downregulates the expression of the longevity transcription factor FOXM1, accelerates osteoclast aging, and ultimately inhibits bone resorption and reverses osteoporosis. Therefore, the nanomaterial that targets and regulates intracellular pH can be applied to the treatment of osteoporosis.

Claims

1. Nanomaterials for targeted regulation of intracellular pH, characterized by: The nanomaterial is a targeted cell membrane nanovesicle encapsulating pH buffer salt; the targeted cell membrane comes from cells that need to regulate the intracellular pH value.

2. The nanomaterial for targeted regulation of intracellular pH according to claim 1, wherein: The pH buffer salt is a phosphate, carbonate, citrate or succinate compound.

3. The nanomaterial for targeted regulation of intracellular pH according to claim 1 or 2, characterized in that: The pH buffer salt is at least one of disodium hydrogen phosphate, dipotassium hydrogen phosphate, diammonium hydrogen phosphate, sodium dihydrogen phosphate, potassium dihydrogen phosphate, ammonium dihydrogen phosphate, sodium bicarbonate, potassium bicarbonate, ammonium bicarbonate, sodium carbonate, potassium carbonate, ammonium carbonate, sodium citrate, potassium citrate, magnesium citrate, ferric citrate, sodium succinate, potassium succinate, and magnesium succinate.

4. The nanomaterial for targeted regulation of intracellular pH according to claim 1, wherein: The target cell membrane is selected from the cell membranes of cells related to the motor system, endocrine system, and digestive system.

5. The nanomaterial for targeted regulation of intracellular pH according to claim 1 or 4, characterized in that: The targeted cell membrane is a chondrocyte membrane, an osteoblast membrane, an osteoclast membrane, a muscle cell membrane, a mesenchymal stem cell membrane, a pancreatic β cell membrane, a pancreatic α cell membrane, an adrenal medullary cell membrane, a thyroid follicular cell membrane, a parathyroid cell membrane, a gastric epithelial cell membrane, a liver cell membrane, a pancreatic acinar cell membrane or an intestinal epithelial cell membrane.

6. The method for preparing a nanomaterial for targeted regulation of intracellular pH according to any one of claims 1, 2, and 4, wherein the target cell membrane is extracted by differential centrifugation, characterized in that: The pH buffer salt is mixed with the targeted cell membrane by ultrasound, and the mixture is filtered and sterilized after low-temperature dialysis to obtain the nanomaterial for targeted regulation of intracellular pH value.

7. The method for preparing a nanomaterial for targeted regulation of intracellular pH according to claim 6, characterized in that: The specific steps include: Step 1: Extract target cell membrane by differential centrifugation The desired targeted cells were resuspended in TM buffer pre-cooled at 4°C, and the cells were crushed by extrusion using a micro-extruder for 40 cycles; the cell lysis solution was mixed with a 1.00 mol / L sucrose solution at a volume ratio of 3:1 to prepare a sucrose-TM buffer mixture with a final concentration of 0.25 mol / L; the sucrose-TM buffer mixture was centrifuged at 2000×g for 10 minutes, and the supernatant was collected; the supernatant was centrifuged at 3000×g for 30 minutes, the supernatant was discarded, and the precipitate was taken; the precipitate was resuspended with a 0.25 mol / L sucrose solution, and centrifuged again at 3000×g for 30 minutes. The resulting precipitate is the desired targeted cell membrane component; The TM buffer comprises mannitol, sucrose, Tris, MgCl2, KCl, PMSF, EDTA-free protease inhibitors, DNAse and RNAse, with a pH of 7.4; wherein the concentration of Tris is 5 to 30 mmol / L, and the concentration of MgCl2 is 1 to 20 mmol / L; Step 2: Preparation of cell membrane biomimetic nanovesicles loaded with pH buffer salt compounds Add 10.00 mL of 0.5 mol / L pH buffered saline solution to the targeted cell membrane, shake and hydrate at 37°C for 30 minutes, dialyze in a dialysis bag for 48 hours, filter through a 0.22 μm sterile filter membrane to obtain the nanomaterial for targeted regulation of intracellular pH value, and store at 4°C in the dark.

8. The use of the nanomaterial for targeted regulation of intracellular pH according to any one of claims 1, 2, and 4, characterized in that: The nanomaterial is used to specifically identify target cells, release regulators, and target and regulate the intracellular pH value of the target cells.

9. The use of the nanomaterial for targeted regulation of intracellular pH as claimed in claim 8, characterized in that: The use of pancreatic beta cell membrane nanovesicles loaded with sodium citrate in the preparation of type 2 diabetes drugs, or the use of intestinal epithelial cell membrane nanovesicles loaded with sodium bicarbonate in the preparation of type 2 inflammatory bowel disease drugs.

10. The use of the nanomaterial for targeted regulation of intracellular pH as claimed in claim 8, characterized in that: Application of precursor osteoclast membrane nanovesicles loaded with disodium hydrogen phosphate in the preparation of osteoporosis drugs.

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