Tetrodotoxin nanoparticles as well as preparation method and application thereof

Abstract

The invention belongs to the field of medicines, and provides tetrodotoxin nanoparticles as well as a preparation method and application thereof. The nanoparticle comprises an inner core and a shell, the inner core comprises an active component and a cosolvent; the active component comprises tetrodotoxin or a pharmaceutically acceptable salt thereof; the shell comprises at least two layers of nanoparticle shells; each layer of nanoparticle shell comprises a polymer matrix layer and a stabilizer layer; a polymer matrix in the polymer matrix layer comprises polylactic acid-glycolic acid copolymer-polyethylene glycol. The nanoparticle provided by the invention is good in stability, can be stored for a long time at room temperature, solves the problem that a tetrodotoxin preparation is unstable at room temperature, and in addition, the nanoparticle provided by the invention is good in slow release effect and can be continuously released within at least 24 hours.

Description

Technical Field

[0001] The present invention relates to the field of drugs, and particularly relates to a tetrodotoxin nanoparticle, a preparation method thereof, and an application thereof. Background Art

[0002] Tetrodotoxin (TTX) is a highly selective sodium ion channel blocker, which has effects such as analgesia, drug detoxification, and anti-arrhythmia. Since TTX is a highly hydrophilic alkaloid small molecule active substance, it is not resistant to acids and alkalis, has a short half-life in the body after intramuscular injection, a low therapeutic index, and poor safety.

[0003] Tetrodotoxin is easily degraded by the influence of temperature in the injection solution. The higher the temperature, the faster the degradation.

[0004] In addition, due to the physical properties of tetrodotoxin itself, the dissolution of tetrodotoxin requires the use of organic acids and their salts or buffer solutions as solubilizers. However, tetrodotoxin will degrade under strong acidic or alkaline conditions.

[0005] Currently developed tetrodotoxin injections cannot be stored for a long time at room temperature, but need to be stored under low temperature conditions of 2 - 8°C, which brings many inconveniences to drug production, storage, transportation, sales, and clinical use.

[0006] In addition, the existing tetrodotoxin injections have poor sustained-release effects and are difficult to achieve long-term sustained release.

[0007] Nanoparticles are often used as drug delivery carriers due to their high bioavailability, high encapsulation rate, controlled release, and low toxicity characteristics, and have advantages such as improving drug efficacy, reducing side effects, and improving patient compliance. However, it has been found through research that tetrodotoxin nanoparticles prepared by conventional methods have the problem of poor sustained-release effects and are difficult to achieve long-term sustained release effects.

[0008] Therefore, there is still an urgent need for a tetrodotoxin pharmaceutical preparation product with good stability and good sustained-release effects. Summary of the Invention

[0009] To solve the above technical problems, the present invention provides the following technical solutions.

[0010] In a first aspect, the present invention provides a nanoparticle with a core-shell structure.

[0011] A nanoparticle with a core-shell structure, which comprises: a core and a shell; The core comprises an active ingredient and a solubilizer; the active ingredient comprises tetrodotoxin or a pharmaceutically acceptable salt thereof; The shell comprises at least two layers of nanoparticle shells; each layer of the nanoparticle shell comprises a polymer matrix layer and a stabilizer layer; The polymer matrix in the polymer matrix layer includes poly(lactic-co-glycolic acid)-polyethylene glycol. In the technical solution of this embodiment, compared with using other polymer matrices, using the poly(lactic-co-glycolic acid)-polyethylene glycol provided by the present invention as the polymer matrix is more conducive to reducing the particle size of the obtained nanoparticles, more conducive to improving the encapsulation efficiency of the obtained nanoparticles, more conducive to improving the stability of the content of tixotrope, the stability of the encapsulation efficiency and the stability of related substances of the obtained nanoparticles, and has unexpected technical effects. Compared with the nanoparticles with a single-layer nanoparticle shell, the present invention uses at least two layers of nanoparticle shells, which is more conducive to improving the encapsulation efficiency of the obtained nanoparticles, more conducive to improving the stability of the content of tixotrope, the stability of the encapsulation efficiency and the stability of related substances of the obtained nanoparticles, and is conducive to prolonging the sustained-release effect of the obtained nanoparticles, and has unexpected technical effects.

[0012] In some embodiments, the co-solvent includes at least one of citric acid, acetic acid, malic acid or tartaric acid. In some embodiments, the co-solvent is preferably citric acid. Compared with using other co-solvents, using the citric acid provided by the present invention as the co-solvent is more conducive to improving the stability of the content of tixotrope and the stability of related substances of the obtained nanoparticles, and has unexpected technical effects.

[0013] In some embodiments, the active ingredient is encapsulated within the innermost layer of the nanoparticle shell.

[0014] In some embodiments, in each layer of the nanoparticle shell, from the inside out, there are a polymer matrix layer and a stabilizer layer in sequence.

[0015] In some embodiments, in each layer of the nanoparticle shell, the stabilizers in the stabilizer layer independently include at least one of polyvinyl alcohol, vitamin E polyethylene glycol succinate or Tween 80. In some embodiments, in each layer of the nanoparticle shell, the stabilizer in the stabilizer layer is preferably polyvinyl alcohol. Compared with using other stabilizers, using the PVA provided by the present invention as the stabilizer is more conducive to improving the encapsulation efficiency of the obtained nanoparticles, more conducive to improving the stability of the content of tixotrope, the stability of the encapsulation efficiency and the stability of related substances of the obtained nanoparticles, and has unexpected technical effects. In some embodiments, in each layer of the nanoparticle shell, the stabilizer in the stabilizer layer is preferably polyvinyl alcohol type 1788. Compared with using other stabilizers, using the polyvinyl alcohol type 1788 provided by the present invention as the stabilizer is more conducive to improving the encapsulation efficiency of the obtained nanoparticles, more conducive to improving the stability of the content of tixotrope, the stability of the encapsulation efficiency and the stability of related substances of the obtained nanoparticles, and has unexpected technical effects.

[0016] In some embodiments, the weight-average molecular weight of poly(lactic-co-glycolic acid)-polyethylene glycol in the innermost nanoparticle shell of the nanoparticles is 10 kDa - 100 kDa. In some embodiments, the weight-average molecular weight of poly(lactic-co-glycolic acid)-polyethylene glycol in the innermost nanoparticle shell of the nanoparticles is 10 kDa, 20 kDa, 30 kDa, 40 kDa, 50 kDa, 60 kDa, 70 kDa, 80 kDa, 90 kDa or 100 kDa. In some embodiments, the weight-average molecular weight of poly(lactic-co-glycolic acid)-polyethylene glycol in the innermost nanoparticle shell of the nanoparticles is preferably 50 kDa. For the weight-average molecular weight of the innermost shell polymer matrix, compared with other weight-average molecular weights, using the weight-average molecular weight of 50 kDa provided by the present invention is more conducive to improving the encapsulation efficiency of the obtained nanoparticles, and is more conducive to improving the stability of the content of tixotroxin, the stability of the encapsulation efficiency and the stability of related substances in the obtained nanoparticles, and has unexpected technical effects.

[0017] In some embodiments, the weight-average molecular weight of poly(lactic-co-glycolic acid)-polyethylene glycol in the other layer nanoparticle shells outside the innermost nanoparticle shell of the nanoparticles is 10 kDa - 100 kDa. In some embodiments, the weight-average molecular weight of poly(lactic-co-glycolic acid)-polyethylene glycol in the other layer nanoparticle shells outside the innermost nanoparticle shell of the nanoparticles is 10 kDa, 20 kDa, 30 kDa, 40 kDa, 50 kDa, 60 kDa, 70 kDa, 80 kDa, 90 kDa or 100 kDa. In some embodiments, the weight-average molecular weight of poly(lactic-co-glycolic acid)-polyethylene glycol in the other layer nanoparticle shells outside the innermost nanoparticle shell of the nanoparticles is preferably 100 kDa. For the weight-average molecular weight of the outer shell polymer matrix, compared with other weight-average molecular weights (such as 10 kDa - 50 kDa), using the weight-average molecular weight of 100 kDa provided by the present invention is more conducive to improving the stability of the content of tixotroxin, the stability of the encapsulation efficiency and the stability of related substances in the obtained nanoparticles, and has unexpected technical effects.

[0018] In some embodiments, the mass ratio of the active ingredient to the polymer matrix in the innermost nanoparticle shell of the nanoparticles is 1:100 to 1:1000. In some embodiments, the mass ratio of the active ingredient to the polymer matrix in the innermost nanoparticle shell of the nanoparticles is 1:100, 1:200, 1:250, 1:300, 1:350, 1:400, 1:500, 1:600, 1:700, 1:800, 1:900, or 1:1000. In some embodiments, the mass ratio of the active ingredient to the polymer matrix in the innermost nanoparticle shell of the nanoparticles is preferably 1:100 - 1:400. In some embodiments, the mass ratio of the active ingredient to the polymer matrix in the innermost nanoparticle shell of the nanoparticles is preferably 1:200 - 1:400.

[0019] In some embodiments, the mass ratio of the active ingredient to the cosolvent is 1:0.3 - 1:3. In some embodiments, the mass ratio of the active ingredient to the cosolvent is 1:0.3, 1:0.5, 1:0.6, 1:0.7, 1:0.8, 1:0.9, 1:1, 1:1.1, 1:1.2, 1:1.3, 1:1.4, 1:1.5, 1:2, 1:2.5, or 1:3. In some embodiments, the mass ratio of the active ingredient to the cosolvent is preferably 1:1.

[0020] In some embodiments, the mass ratio of the active ingredient to the polymer matrix in the other nanoparticle shells outside the innermost nanoparticle shell of the nanoparticles is 1:200 to 1:2000. In some embodiments, the mass ratio of the active ingredient to the polymer matrix in the other nanoparticle shells outside the innermost nanoparticle shell of the nanoparticles is 1:200, 1:300, 1:400, 1:500, 1:600, 1:700, 1:800, 1:900, 1:1000, 1:1100, 1:1200, 1:1300, 1:1400, 1:1500, 1:1600, 1:1700, 1:1800, 1:1900, or 1:2000. In some embodiments, the mass ratio of the active ingredient to the polymer matrix in the other nanoparticle shells outside the innermost nanoparticle shell of the nanoparticles is preferably 1:200 - 1:800. In some embodiments, the mass ratio of the active ingredient to the polymer matrix in the other nanoparticle shells outside the innermost nanoparticle shell of the nanoparticles is preferably 1:400 - 1:800.

[0021] In some embodiments, in the innermost nanoparticle shell of the nanoparticles, the mass ratio of the polymer matrix to the stabilizer is from 1:0.25 to 1:5. In some embodiments, in the innermost nanoparticle shell of the nanoparticles, the mass ratio of the polymer matrix to the stabilizer is 1:0.25, 1:0.5, 1:0.75, 1:1, 1:1.5, 1:2, 1:2.5, 1:3, 1:3.5, 1:4, 1:4.5 or 1:5. In some embodiments, in the innermost nanoparticle shell of the nanoparticles, the mass ratio of the polymer matrix to the stabilizer is preferably from 1:0.5 to 1:2.5. In some embodiments, in the innermost nanoparticle shell of the nanoparticles, the mass ratio of the polymer matrix to the stabilizer is preferably from 1:1 to 1:2.5.

[0022] In some embodiments, in the other nanoparticle shells outside the innermost nanoparticle shell of the nanoparticles, the mass ratio of the polymer matrix to the stabilizer is greater than 0:25 to less than or equal to 200:25. In some embodiments, in the other nanoparticle shells outside the innermost nanoparticle shell of the nanoparticles, the mass ratio of the polymer matrix to the stabilizer is 1:25, 5:25, 10:25, 15:25, 20:25, 25:25, 30:25, 35:25, 40:25, 45:25, 50:25, 60:25, 70:25, 80:25, 90:25, 100:25, 110:25, 120:25, 130:25, 140:25, 150:25, 160:25, 170:25, 180:25, 190:25 or 200:25. In some embodiments, in the other nanoparticle shells outside the innermost nanoparticle shell of the nanoparticles, the mass ratio of the polymer matrix to the stabilizer is preferably from 20:25 to 40:25.

[0023] In some embodiments, the nanoparticles comprise two nanoparticle shells.

[0024] In some embodiments, the proportion of each component of the nanoparticles is as follows: the mass ratio of the active ingredient to the co-solvent is 1:1; the mass ratio of the active ingredient to the polymer matrix in the innermost nanoparticle shell of the nanoparticles is from 1:100 to 1:400; in the innermost nanoparticle shell of the nanoparticles, the mass ratio of the polymer matrix to the stabilizer is from 1:0.5 to 1:2.5 or from 1:1 to 1:2.5; the mass ratio of the active ingredient to the polymer matrix in the other nanoparticle shells outside the innermost nanoparticle shell of the nanoparticles is from 1:200 to 1:800; in the other nanoparticle shells outside the innermost nanoparticle shell of the nanoparticles, the mass ratio of the polymer matrix to the stabilizer is from 20:25 to 40:25.

[0025] In some embodiments, the ratios of the components of the nanoparticles are as follows: the mass ratio of the active ingredient to the cosolvent is 1:1; the mass ratio of the active ingredient to the polymer matrix in the innermost nanoparticle shell of the nanoparticles is 1:200 - 1:400; in the innermost nanoparticle shell of the nanoparticles, the mass ratio of the polymer matrix to the stabilizer is 1:1 - 1:2.5; the mass ratio of the active ingredient to the polymer matrix in the other nanoparticle shells outside the innermost nanoparticle shell of the nanoparticles is 1:400 - 1:800; in the other nanoparticle shells outside the innermost nanoparticle shell of the nanoparticles, the mass ratio of the polymer matrix to the stabilizer is 20:25 - 40:25.

[0026] In some embodiments, the ratios of the components of the nanoparticles are as follows: the mass ratio of the active ingredient to the cosolvent is 1:1; the mass ratio of the active ingredient to the polymer matrix in the innermost nanoparticle shell of the nanoparticles is 1:200; in the innermost nanoparticle shell of the nanoparticles, the mass ratio of the polymer matrix to the stabilizer is 1:2.5; the mass ratio of the active ingredient to the polymer matrix in the other nanoparticle shells outside the innermost nanoparticle shell of the nanoparticles is 1:800; in the other nanoparticle shells outside the innermost nanoparticle shell of the nanoparticles, the mass ratio of the polymer matrix to the stabilizer is 40:25.

[0027] In some embodiments, the ratios of the components of the nanoparticles are as follows: the mass ratio of the active ingredient to the cosolvent is 1:1; the mass ratio of the active ingredient to the polymer matrix in the innermost nanoparticle shell of the nanoparticles is 1:200; in the innermost nanoparticle shell of the nanoparticles, the mass ratio of the polymer matrix to the stabilizer is 1:2; the mass ratio of the active ingredient to the polymer matrix in the other nanoparticle shells outside the innermost nanoparticle shell of the nanoparticles is 1:800; in the other nanoparticle shells outside the innermost nanoparticle shell of the nanoparticles, the mass ratio of the polymer matrix to the stabilizer is 40:25.

[0028] In some embodiments, the ratios of the components of the nanoparticles are as follows: the mass ratio of the active ingredient to the cosolvent is 1:1; the mass ratio of the active ingredient to the polymer matrix in the innermost nanoparticle shell of the nanoparticles is 1:400; in the innermost nanoparticle shell of the nanoparticles, the mass ratio of the polymer matrix to the stabilizer is 1:1.25; the mass ratio of the active ingredient to the polymer matrix in the other nanoparticle shells outside the innermost nanoparticle shell of the nanoparticles is 1:400; in the other nanoparticle shells outside the innermost nanoparticle shell of the nanoparticles, the mass ratio of the polymer matrix to the stabilizer is 20:25.

[0029] In some embodiments, the ratios of the components of the nanoparticles are as follows: the mass ratio of the active ingredient to the cosolvent is 1:1; the mass ratio of the active ingredient to the polymer matrix in the innermost nanoparticle shell of the nanoparticles is 1:400; in the innermost nanoparticle shell of the nanoparticles, the mass ratio of the polymer matrix to the stabilizer is 1:2.5; the mass ratio of the active ingredient to the polymer matrix in the other nanoparticle shells outside the innermost nanoparticle shell of the nanoparticles is 1:800; in the other nanoparticle shells outside the innermost nanoparticle shell of the nanoparticles, the mass ratio of the polymer matrix to the stabilizer is 40:25.

[0030] In a second aspect, the present invention provides a method for preparing the nanoparticles described in the first aspect.

[0031] A method for preparing the nanoparticles described in the first aspect, which comprises the following steps: (S1) Preparation of primary emulsion (W / O): Dissolve the active ingredient in an aqueous solution containing a cosolvent to obtain Solution 1, and then add Solution 1 to an organic solvent containing a polymer matrix, and homogenize at high speed to form a primary emulsion (W / O); (S2) Preparation of multiple emulsion (W / O / W): Add the above primary emulsion (W / O) to an aqueous solution containing a stabilizer, homogenize at high speed to form a multiple emulsion (W / O / W), add water and stir to solidify, centrifuge to obtain Solid A, wash the obtained Solid A, and centrifuge to obtain Solid B; (S3) Formation of the second polymer matrix layer: Add Solid B obtained in step (S2) to an organic solvent containing a polymer matrix, and homogenize at high speed to obtain Emulsion 2; (S4) Formation of double-shell nanoparticles: Add Emulsion 2 obtained in step (S3) to an aqueous solution containing a stabilizer, homogenize at high speed to obtain Emulsion 3, add water and stir to solidify, centrifuge to obtain Solid C, wash the obtained Solid C, and centrifuge to obtain Solid D, and lyophilize Solid D to obtain double-shell nanoparticles.

[0032] In some embodiments, the feeding mass ratio of the active ingredient to the polymer matrix in step (S1) is 1:100 to 1:1000. In some embodiments, the feeding mass ratio of the active ingredient to the polymer matrix in step (S1) is 1:100, 1:200, 1:250, 1:300, 1:350, 1:400, 1:500, 1:600, 1:700, 1:800, 1:900 or 1:1000. In some embodiments, the feeding mass ratio of the active ingredient to the polymer matrix in step (S1) is preferably 1:100 - 1:400. In some embodiments, the feeding mass ratio of the active ingredient to the polymer matrix in step (S1) is preferably 1:200 - 1:400.

[0033] In some embodiments, the feeding mass ratio of the active ingredient to the cosolvent is 1:0.3 - 1:3. In some embodiments, the feeding mass ratio of the active ingredient to the cosolvent is 1:0.3, 1:0.5, 1:0.6, 1:0.7, 1:0.8, 1:0.9, 1:1, 1:1.1, 1:1.2, 1:1.3, 1:1.4, 1:1.5, 1:2, 1:2.5 or 1:3. In some embodiments, the feeding mass ratio of the active ingredient to the cosolvent is preferably 1:1.

[0034] In some embodiments, the feeding mass ratio of the active ingredient to the polymer matrix in step (S3) is 1:200 to 1:2000. In some embodiments, the feeding mass ratio of the active ingredient to the polymer matrix in step (S3) is 1:200, 1:300, 1:400, 1:500, 1:600, 1:700, 1:800, 1:900, 1:1000, 1:1100, 1:1200, 1:1300, 1:1400, 1:1500, 1:1600, 1:1700, 1:1800, 1:1900 or 1:2000. In some embodiments, the feeding mass ratio of the active ingredient to the polymer matrix in step (S3) is preferably 1:200 - 1:800. In some embodiments, the feeding mass ratio of the active ingredient to the polymer matrix in step (S3) is preferably 1:400 - 1:800.

[0035] In some embodiments, the feeding mass ratio of the polymer matrix in step (S1) to the stabilizer in step (S2) is 1:0.25 to 1:5. In some embodiments, the feeding mass ratio of the polymer matrix in step (S1) to the stabilizer in step (S2) is 1:0.25, 1:0.5, 1:0.75, 1:1, 1:1.5, 1:2, 1:2.5, 1:3, 1:3.5, 1:4, 1:4.5 or 1:5. In some embodiments, the feeding mass ratio of the polymer matrix in step (S1) to the stabilizer in step (S2) is preferably 1:0.5 to 1:2.5. In some embodiments, the feeding mass ratio of the polymer matrix in step (S1) to the stabilizer in step (S2) is preferably 1:1 to 1:2.5.

[0036] In some embodiments, the feeding mass ratio of the polymer matrix in step (S3) to the stabilizer in step (S4) is greater than 0:25 to less than or equal to 200:25. In some embodiments, the feeding mass ratio of the polymer matrix in step (S3) to the stabilizer in step (S4) is 1:25, 5:25, 10:25, 15:25, 20:25, 25:25, 30:25, 35:25, 40:25, 45:25, 50:25, 60:25, 70:25, 80:25, 90:25, 100:25, 110:25, 120:25, 130:25, 140:25, 150:25, 160:25, 170:25, 180:25, 190:25 or 200:25. In some embodiments, the feeding mass ratio of the polymer matrix in step (S3) to the stabilizer in step (S4) is preferably 20:25 to 40:25.

[0037] In some embodiments, the feeding ratio of each component is as follows: the feeding mass ratio of the active ingredient to the cosolvent is 1:1; the feeding mass ratio of the active ingredient to the polymer matrix in step (S1) is 1:100 - 1:400; the feeding mass ratio of the polymer matrix in step (S1) to the stabilizer in step (S2) is 1:0.5 to 1:2.5; the feeding mass ratio of the active ingredient to the polymer matrix in step (S3) is 1:200 - 1:800; the feeding mass ratio of the polymer matrix in step (S3) to the stabilizer in step (S4) is 20:25 to 40:25.

[0038] In some embodiments, the feeding ratios of the components are as follows: the feeding mass ratio of the active ingredient to the cosolvent is 1:1; the feeding mass ratio of the active ingredient to the polymer matrix in step (S1) is 1:100 - 1:400; the feeding mass ratio of the polymer matrix in step (S1) to the stabilizer in step (S2) is 1:1 - 1:2.5; the feeding mass ratio of the active ingredient to the polymer matrix in step (S3) is 1:200 - 1:800; the feeding mass ratio of the polymer matrix in step (S3) to the stabilizer in step (S4) is 20:25 - 40:25.

[0039] In some embodiments, the feeding ratios of the components are as follows: the feeding mass ratio of the active ingredient to the cosolvent is 1:1; the feeding mass ratio of the active ingredient to the polymer matrix in step (S1) is 1:200 - 1:400; the feeding mass ratio of the polymer matrix in step (S1) to the stabilizer in step (S2) is 1:1 - 1:2.5; the feeding mass ratio of the active ingredient to the polymer matrix in step (S3) is 1:400 - 1:800; the feeding mass ratio of the polymer matrix in step (S3) to the stabilizer in step (S4) is 20:25 - 40:25.

[0040] In some embodiments, the feeding ratios of the components are as follows: the feeding mass ratio of the active ingredient to the cosolvent is 1:1; the feeding mass ratio of the active ingredient to the polymer matrix in step (S1) is 1:200; the feeding mass ratio of the polymer matrix in step (S1) to the stabilizer in step (S2) is 1:2.5; the feeding mass ratio of the active ingredient to the polymer matrix in step (S3) is 1:800; the feeding mass ratio of the polymer matrix in step (S3) to the stabilizer in step (S4) is 40:25.

[0041] In some embodiments, the feeding ratios of the components are as follows: the feeding mass ratio of the active ingredient to the cosolvent is 1:1; the feeding mass ratio of the active ingredient to the polymer matrix in step (S1) is 1:200; the feeding mass ratio of the polymer matrix in step (S1) to the stabilizer in step (S2) is 1:2; the feeding mass ratio of the active ingredient to the polymer matrix in step (S3) is 1:800; the feeding mass ratio of the polymer matrix in step (S3) to the stabilizer in step (S4) is 40:25.

[0042] In some embodiments, the feeding ratios of the components are as follows: the feeding mass ratio of the active ingredient to the cosolvent is 1:1; the feeding mass ratio of the active ingredient to the polymer matrix in step (S1) is 1:400; the feeding mass ratio of the polymer matrix in step (S1) to the stabilizer in step (S2) is 1:1.25; the feeding mass ratio of the active ingredient to the polymer matrix in step (S3) is 1:400; the feeding mass ratio of the polymer matrix in step (S3) to the stabilizer in step (S4) is 20:25.

[0043] In some embodiments, the feeding ratios of the components are as follows: the feeding mass ratio of the active ingredient to the cosolvent is 1:1; the feeding mass ratio of the active ingredient to the polymer matrix in step (S1) is 1:400; the feeding mass ratio of the polymer matrix in step (S1) to the stabilizer in step (S2) is 1:2.5; the feeding mass ratio of the active ingredient to the polymer matrix in step (S3) is 1:800; the feeding mass ratio of the polymer matrix in step (S3) to the stabilizer in step (S4) is 40:25.

[0044] In some embodiments, the organic solvents in step (S1) or step (S3) are each independently selected from dichloromethane.

[0045] In some embodiments, the concentration of the cosolvent in the aqueous solution containing the cosolvent in step (S1) is 0.1 wt% - 1 wt%. In some embodiments, the concentration of the cosolvent in the aqueous solution containing the cosolvent in step (S1) is 0.1 wt%, 0.2 wt%, 0.3 wt%, 0.4 wt%, 0.5 wt%, 0.6 wt%, 0.7 wt%, 0.8 wt%, 0.9 wt% or 1 wt%. In some embodiments, the concentration of the cosolvent in the aqueous solution containing the cosolvent in step (S1) is preferably 0.1 wt%.

[0046] In some embodiments, in the organic solvent containing the polymer matrix in step (S1), 25 mg - 300 mg of the polymer matrix is fed per 1 ml of the organic solvent. In some embodiments, in the organic solvent containing the polymer matrix in step (S1), 25 mg, 50 mg, 75 mg, 100 mg, 150 mg, 200 mg, 250 mg or 300 mg of the polymer matrix is fed per 1 ml of the organic solvent. In some embodiments, in the organic solvent containing the polymer matrix in step (S1), 100 mg of the polymer matrix is fed per 1 ml of the organic solvent.

[0047] In some embodiments, in the aqueous solution containing a stabilizer in step (S2), the concentration of the stabilizer is 0.5 wt% - 2 wt%. In some embodiments, in the aqueous solution containing a stabilizer in step (S2), the concentration of the stabilizer is 0.5 wt%, 1 wt%, 1.5 wt% or 2 wt%. In some embodiments, in the aqueous solution containing a stabilizer in step (S2), the concentration of the stabilizer is preferably 1 wt%.

[0048] In some embodiments, in the operation of adding water and stirring for curing in step (S2), the volume ratio of the multiple emulsion (W / O / W) to water is greater than 0:2 and less than or equal to 1:2. In some embodiments, in the operation of adding water and stirring for curing in step (S2), the volume ratio of the multiple emulsion (W / O / W) to water is 0.1:2, 0.2:2, 0.3:2, 0.4:2, 0.5:2, 0.6:2, 0.7:2, 0.8:2, 0.9:2 or 1:2. In some embodiments, in the operation of adding water and stirring for curing in step (S2), the volume ratio of the multiple emulsion (W / O / W) to water is preferably 1:2.

[0049] In some embodiments, the time for adding water and stirring for curing in step (S2) is 0 - 24 h. In some embodiments, the time for adding water and stirring for curing in step (S2) is 0 h, 5 h, 10 h, 12 h, 15 h, 20 h, 21 h, 22 h, 23 h or 24 h. In some embodiments, the time for adding water and stirring for curing in step (S2) is 24 h.

[0050] In some embodiments, the rotation speed of any centrifugation in step (S2) is independently selected from 5000 rpm - 15000 rpm. In some embodiments, the rotation speed of any centrifugation in step (S2) is independently selected from 5000 rpm, 6000 rpm, 7000 rpm, 8000 rpm, 9000 rpm, 10000 rpm, 11000 rpm, 12000 rpm, 13000 rpm, 14000 rpm or 15000 rpm. In some embodiments, the rotation speed of any centrifugation in step (S2) is 10000 rpm.

[0051] In some embodiments, the centrifugation time of any centrifugation in step (S2) is independently selected from 5 min - 30 min. In some embodiments, the centrifugation time of any centrifugation in step (S2) is 5 min, 10 min, 15 min, 20 min, 25 min or 30 min. In some embodiments, the centrifugation time of any centrifugation in step (S2) is 15 min.

[0052] In some embodiments, the cleaning in step (S2) is carried out with water.

[0053] In some embodiments, in the organic solvent of the polymer matrix in step (S3), 25 mg - 300 mg of the polymer matrix is fed per 1 ml of the organic solvent. In some embodiments, in the organic solvent of the polymer matrix in step (S3), 25 mg, 50 mg, 75 mg, 100 mg, 150 mg, 200 mg, 250 mg or 300 mg of the polymer matrix is fed per 1 ml of the organic solvent. In some embodiments, in the organic solvent of the polymer matrix in step (S3), 100 mg of the polymer matrix is fed per 1 ml of the organic solvent.

[0054] In some embodiments, in the aqueous solution containing a stabilizer in step (S4), the concentration of the stabilizer is 0.5 wt% - 2 wt%. In some embodiments, in the aqueous solution containing a stabilizer in step (S4), the concentration of the stabilizer is 0.5 wt%, 1 wt%, 1.5 wt% or 2 wt%. In some embodiments, in the aqueous solution containing a stabilizer in step (S4), the concentration of the stabilizer is 1 wt%.

[0055] In some embodiments, the weight-average molecular weight of the polymer matrix in step (S1) is 10 kDa - 100 kDa. In some embodiments, the weight-average molecular weight of the polymer matrix in step (S1) is 10 kDa, 20 kDa, 30 kDa, 40 kDa, 45 kDa, 50 kDa, 55 kDa, 60 kDa, 70 kDa, 80 kDa, 90 kDa or 100 kDa. In some embodiments, the weight-average molecular weight of the polymer matrix in step (S1) is preferably 50 kDa.

[0056] In some embodiments, the weight-average molecular weight of the polymer matrix in step (S3) is 10 - 100 kDa. In some embodiments, the weight-average molecular weight of the polymer matrix in step (S3) is 10 kDa, 20 kDa, 30 kDa, 40 kDa, 45 kDa, 50 kDa, 55 kDa, 60 kDa, 70 kDa, 80 kDa, 90 kDa or 100 kDa. In some embodiments, the weight-average molecular weight of the polymer matrix in step (S3) is preferably 100 kDa.

[0057] In some embodiments, the rotation speed of the high-speed homogenization in step (S1) is 2000 rpm - 10000 rpm. In some embodiments, the rotation speed of the high-speed homogenization in step (S1) is 2000 rpm, 3000 rpm, 4000 rpm, 5000 rpm, 6000 rpm, 7000 rpm, 8000 rpm, 9000 rpm or 10000 rpm. In some embodiments, the rotation speed of the high-speed homogenization in step (S1) is 10000 rpm.

[0058] In some embodiments, the time of the high-speed homogenization in step (S1) is 2 min - 5 min. In some embodiments, the time of the high-speed homogenization in step (S1) is 2 min, 3 min, 4 min or 5 min. In some embodiments, the time of the high-speed homogenization in step (S1) is 5 min.

[0059] In some embodiments, the rotation speed of the high-speed homogenization in step (S2) is 2000 rpm - 10000 rpm. In some embodiments, the rotation speed of the high-speed homogenization in step (S2) is 2000 rpm, 3000 rpm, 4000 rpm, 5000 rpm, 6000 rpm, 7000 rpm, 8000 rpm, 9000 rpm or 10000 rpm. In some embodiments, the rotation speed of the high-speed homogenization in step (S2) is 10000 rpm.

[0060] In some embodiments, the time of the high-speed homogenization in step (S2) is 2 min - 5 min. In some embodiments, the time of the high-speed homogenization in step (S2) is 2 min, 3 min, 4 min or 5 min. In some embodiments, the time of the high-speed homogenization in step (S2) is 5 min.

[0061] In some embodiments, the rotation speed of the high-speed homogenization in step (S3) is 2000 rpm - 10000 rpm. In some embodiments, the rotation speed of the high-speed homogenization in step (S3) is 2000 rpm, 3000 rpm, 4000 rpm, 5000 rpm, 6000 rpm, 7000 rpm, 8000 rpm, 9000 rpm or 10000 rpm. In some embodiments, the rotation speed of the high-speed homogenization in step (S3) is 7000 rpm.

[0062] In some embodiments, the time for high-speed homogenization in step (S3) is 2 min - 5 min. In some embodiments, the time for high-speed homogenization in step (S3) is 2 min, 3 min, 4 min, or 5 min. In some embodiments, the time for high-speed homogenization in step (S3) is 5 min.

[0063] In some embodiments, the rotation speed for high-speed homogenization in step (S4) is 2000 rpm - 10000 rpm. In some embodiments, the rotation speed for high-speed homogenization in step (S4) is 2000 rpm, 3000 rpm, 4000 rpm, 5000 rpm, 6000 rpm, 7000 rpm, 8000 rpm, 9000 rpm, or 10000 rpm. In some embodiments, the rotation speed for high-speed homogenization in step (S4) is 7000 rpm.

[0064] In some embodiments, the time for high-speed homogenization in step (S4) is 2 min - 5 min. In some embodiments, the time for high-speed homogenization in step (S4) is 2 min, 3 min, 4 min, or 5 min. In some embodiments, the time for high-speed homogenization in step (S4) is 5 min.

[0065] In some embodiments, in the water-adding stirring and curing operation of step (S4), the volume ratio of emulsion 3 to water is greater than 0:2 and less than or equal to 1:2. In some embodiments, in the water-adding stirring and curing operation of step (S4), the volume ratio of emulsion 3 to water is 0.1:2, 0.2:2, 0.3:2, 0.4:2, 0.5:2, 0.6:2, 0.7:2, 0.8:2, 0.9:2, or 1:2. In some embodiments, in the water-adding stirring and curing operation of step (S4), the volume ratio of emulsion 3 to water is 1:2.

[0066] In some embodiments, the time for water-adding stirring and curing in step (S4) is 0 - 24 h. In some embodiments, the time for water-adding stirring and curing in step (S4) is 0 h, 5 h, 10 h, 12 h, 15 h, 20 h, 21 h, 22 h, 23 h, or 24 h. In some embodiments, the time for water-adding stirring and curing in step (S4) is 24 h.

[0067] In some embodiments, the centrifugation speed of any centrifugation in step (S4) is independently selected from 5000 rpm - 15000 rpm. In some embodiments, the centrifugation speed of any centrifugation in step (S4) is independently selected from 5000 rpm, 6000 rpm, 7000 rpm, 8000 rpm, 9000 rpm, 10000 rpm, 11000 rpm, 12000 rpm, 13000 rpm, 14000 rpm or 15000 rpm. In some embodiments, the centrifugation speed of any centrifugation in step (S4) is 10000 rpm.

[0068] In some embodiments, the centrifugation time of any centrifugation in step (S4) is independently selected from 5 min - 30 min. In some embodiments, the centrifugation time of any centrifugation in step (S4) is independently selected from 5 min, 10 min, 15 min, 20 min, 25 min or 30 min. In some embodiments, the centrifugation time of any centrifugation in step (S4) is 15 min.

[0069] In some embodiments, the washing in step (S4) is washing with water.

[0070] In some embodiments, the encapsulation efficiency of the nanoparticles is greater than or equal to 40%. In some embodiments, the encapsulation efficiency of the nanoparticles is 40% - 80%. In some embodiments, the encapsulation efficiency of the nanoparticles is 40%, 41%, 42%, 43%, 44%, 45%, 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75% or 80%.

[0071] In some embodiments, the particle size of the nanoparticles is ≤300 nm. In some embodiments, the particle size of the nanoparticles is ≤250 nm. In some embodiments, the particle size of the nanoparticles is 150 nm - 250 nm. In some embodiments, the particle size of the nanoparticles is 180 nm - 220 nm. In some embodiments, the particle size of the nanoparticles is 180 nm, 185 nm, 190 nm, 195 nm, 200 nm, 205 nm, 210 nm, 215 nm or 220 nm.

[0072] In a third aspect, the present invention provides a pharmaceutical composition.

[0073] In some embodiments, a pharmaceutical composition comprises the nanoparticles described in the first aspect or the nanoparticles obtained by the method described in the second aspect.

[0074] In some embodiments, a pharmaceutical composition comprises the nanoparticles described in the first aspect or the nanoparticles obtained by the method described in the second aspect, and a pharmaceutically acceptable excipient or carrier.

[0075] In some embodiments, the encapsulation efficiency of the nanoparticles in the pharmaceutical composition is greater than or equal to 40%. In some embodiments, the encapsulation efficiency of the nanoparticles in the pharmaceutical composition is 40% - 80%. In some embodiments, the encapsulation efficiency of the nanoparticles in the pharmaceutical composition is 40%, 41%, 42%, 43%, 44%, 45%, 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75% or 80%.

[0076] In a fourth aspect, the present invention provides a pharmaceutical preparation.

[0077] A pharmaceutical preparation comprises the nanoparticles described in the first aspect, or the nanoparticles obtained by the method described in the second aspect, or the pharmaceutical composition described in the third aspect.

[0078] In some embodiments, the pharmaceutical preparation is an injection; In some embodiments, the pharmaceutical preparation is a sustained-release injection or a controlled-release injection.

[0079] In a fifth aspect, the present invention provides an application of the aforementioned nanoparticles, the nanoparticles obtained by the aforementioned method, the aforementioned pharmaceutical composition or the aforementioned pharmaceutical composition.

[0080] The application of the nanoparticles described in the first aspect, the nanoparticles obtained by the method described in the second aspect, the pharmaceutical composition described in the third aspect or the pharmaceutical preparation described in the fourth aspect in the preparation of a drug for analgesia, drug detoxification and / or antiarrhythmia.

[0081] Advantageous Effects Compared with the prior art, at least one of the following advantageous effects is achieved in a certain embodiment of the present invention: (1) For the type of the innermost shell polymer matrix, compared with using other polymer matrices (such as DSPE-PEG (50k), DPPE-PEG (50k), or DMPE-PEG (50k)), using the PEG-PLGA (50k) provided by the present invention is more conducive to reducing the particle size of the obtained nanoparticles, more conducive to improving the encapsulation efficiency of the obtained nanoparticles, more conducive to improving the content stability, encapsulation efficiency stability, and related substance stability of the obtained nanoparticles with telithromycin, and has unexpected technical effects.

[0082] (2) For the weight average molecular weight of the innermost shell polymer matrix, compared with using other weight average molecular weights (such as 10 kDa to 30 kDa, or 100 kDa in Comparative Example 7), using the weight average molecular weight (such as 50 kDa) provided by the present invention is more conducive to improving the encapsulation efficiency of the obtained nanoparticles, more conducive to improving the content stability, encapsulation efficiency stability, and related substance stability of the obtained nanoparticles with telithromycin, and has unexpected technical effects.

[0083] (3) For the type of the outer shell polymer matrix, compared with using other polymer matrices (such as chitosan (100k) or PLGA (100k)), using the PEG-PLGA (100k) provided by the present invention is more conducive to reducing the particle size of the obtained nanoparticles, more conducive to improving the encapsulation efficiency of the obtained nanoparticles, more conducive to improving the content stability, encapsulation efficiency stability, and related substance stability of the obtained nanoparticles with telithromycin, and has unexpected technical effects.

[0084] (4) For the weight average molecular weight of the outer shell polymer matrix, compared with using other weight average molecular weights (such as 10 kDa to 50 kDa), using the weight average molecular weight (such as 100 kDa) provided by the present invention is more conducive to improving the content stability, encapsulation efficiency stability, and related substance stability of the obtained nanoparticles with telithromycin, and has unexpected technical effects.

[0085] (5) For the type of the stabilizer, compared with using other stabilizers (such as polyethylene glycol vitamin E succinate or Tween 80), using PVA1788 provided by the present invention is more conducive to improving the encapsulation efficiency of the obtained nanoparticles, more conducive to improving the content stability, encapsulation efficiency stability, and related substance stability of the obtained nanoparticles with telithromycin, and has unexpected technical effects.

[0086] (6) For the type of the co-solvent, compared with using other co-solvents (such as malic acid, tartaric acid, or acetic acid), using citric acid provided by the present invention is more conducive to improving the content stability and related substance stability of the obtained nanoparticles with telithromycin, and has unexpected technical effects.

[0087] (7) The nanoparticles provided by the present invention have a high encapsulation efficiency, small particle size, small PDI (high particle size uniformity), good stability of the active ingredient content, encapsulation efficiency, particle size, and related substances, and have unexpected technical effects.

[0088] (8) The nanoparticles provided by the present invention have a good sustained-release effect and can be continuously and stably released for at least 24 hours.

[0089] Term Explanation In the description of the present invention, the meaning of "a plurality" is two or more, unless otherwise specifically defined.

[0090] The term "room temperature" refers to the ambient temperature, which means the temperature is about 10°C to about 30°C, or about 20°C to 30°C, or about 25°C.

[0091] The term "wt%" represents mass percentage.

[0092] In the description of this specification, the description with reference to terms such as "one embodiment", "some embodiments", "example", "specific example", or "some examples" means that the specific features, structures, materials, or characteristics described in connection with the embodiment or example are included in at least one embodiment or example of the present invention. In this specification, the schematic representations of the above terms do not necessarily refer to the same embodiment or example. Moreover, the specific features, structures, materials, or characteristics described can be combined in a suitable manner in any one or more embodiments or examples. In addition, without contradiction, those skilled in the art can combine and combine the different embodiments or examples described in this specification and the features of different embodiments or examples.

[0093] In the following content, all the numbers disclosed herein are approximate values, whether or not words such as "about" or "approximately" are used. There may be differences of 1%, 2%, 5%, 7%, 8%, 10%, 15%, or 20% in the numerical value of each number. Whenever a number with an N value is disclosed, any number with a value of N+ / -1%, N+ / -2%, N+ / -3%, N+ / -5%, N+ / -7%, N+ / -8%, N+ / -10%, N+ / -15% or N+ / -20% will be explicitly disclosed, where "+ / -" means plus or minus.

[0094] The term "P < 0.001" indicates that the result has extremely significant or highly significant statistical significance, indicating that there is a probability of more than 99.9% that the differences or relationships reflected by the sample data are almost impossible to be randomly generated and are very reliable and significant.

[0095] The term "P < 0.01" represents a highly significant statistical result, meaning that there is a probability of more than 99% that the differences or relationships reflected in the sample data are not caused by random factors, and the reliability of the result is relatively high.

[0096] The term "P < 0.05" indicates a significant statistical result, suggesting that the differences or relationships reflected in the sample data are unlikely to be caused by random errors, and there is a probability of more than 95% that there are real differences or relationships.

[0097] "rpm" represents the rotational speed unit "revolutions per minute".

[0098] "PEG-PLGA" represents "poly(lactic-co-glycolic acid)-polyethylene glycol".

[0099] "PLGA(30k)" represents "poly(lactic-co-glycolic acid)" with a weight-average molecular weight of 30 kDa.

[0100] "PLGA(100k)" represents "poly(lactic-co-glycolic acid)" with a weight-average molecular weight of 100 kDa. "PEG-PLGA(10k)" represents "poly(lactic-co-glycolic acid)-polyethylene glycol" with a weight-average molecular weight of 10 kDa.

[0101] "PEG-PLGA(30k)" represents "poly(lactic-co-glycolic acid)-polyethylene glycol" with a weight-average molecular weight of 30 kDa.

[0102] "PEG-PLGA(50k)" represents "poly(lactic-co-glycolic acid)-polyethylene glycol" with a weight-average molecular weight of 50 kDa.

[0103] "PEG-PLGA(100k)" represents "poly(lactic-co-glycolic acid)-polyethylene glycol" with a weight-average molecular weight of 100 kDa.

[0104] "PVA" represents polyvinyl alcohol.

[0105] DSPE-PEG(50k) represents 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-polyethylene glycol with a weight-average molecular weight of 50 kDa.

[0106] DPPE-PEG(50k) represents 1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine-polyethylene glycol with a weight-average molecular weight of 50 kDa.

[0107] DMPE-PEG(50k) represents 1,2-dimyristoyl-sn-glycero-3-phosphoethanolamine-polyethylene glycol with a weight-average molecular weight of 50 kDa.

[0108] Chitosan (100k) represents chitosan with a weight-average molecular weight of 100 kDa.

[0109] "Parts by weight" is a unit used to represent the relative weight ratio of substances. It expresses the weights of various substances according to a certain proportional relationship without referring to specific weight values. For example, in a prescription, if substance A is 5 parts by weight and substance B is 3 parts by weight, it means the weight ratio of A to B is 5:3; if the actual weight of substance A is 50 grams, then according to the ratio, the weight of substance B is 30 grams; regardless of the actual weight, the weight ratio between them always remains 5:3.

[0110] "PDI" represents the Polydispersity Index.

[0111] "Polyvinyl alcohol type 1788" or "PVA1788" means the same thing. Here, "17" in "1788" indicates that the degree of polymerization of this polyvinyl alcohol is about 1700, and "88" indicates that the degree of alcoholysis is about 88%. The degree of alcoholysis refers to the content of hydroxyl groups in the polyvinyl alcohol molecule. Detailed implementation modes

[0112] To enable those skilled in the art to better understand the technical solutions of the present invention, some non-limiting embodiments are further disclosed below for further detailed description of the present invention.

[0113] The determination of the content of tixadorine nanoparticles and related substances is as follows: It is slightly modified on the basis of the Marine Industry Standard of the People's Republic of China HY / T 216-2017 "Detection Method for Tetrodotoxin", specifically as follows: Chromatographic conditions and system suitability test: Use octadecylsilane-bonded silica gel as the filler; a phosphate buffer solution of sodium octanesulfonate (weigh 17.91 g of disodium hydrogen phosphate, 7.80 g of sodium dihydrogen phosphate, and 0.54 g of sodium octanesulfonate, add pure water to dissolve to 1000 ml, shake well, filter, and obtain) as the mobile phase; the flow rate is 0.3 ml / min; the excitation wavelength is 365 nm; the emission wavelength is 510 nm; the post-column derivatizing agent is 4 mol / L sodium hydroxide solution; the flow rate of the derivatizing agent is 0.3 ml / min; the post-column derivatization temperature is 110 °C; the injection volume is 50 μl. The number of theoretical plates calculated based on the tixadorine peak is not less than 2000.

[0114] Assay method: Weigh about 100 μg of tixotroxin nanoparticles containing tixotroxin, place it in a 10 ml volumetric flask, add an appropriate amount of dichloromethane and ultrasonically demulsify it, then add pure water to dilute to the mark, shake well, take an appropriate amount and centrifuge, take the supernatant and filter it through a 0.22 μm microporous filter membrane as the test solution. Inject 20 μl of the test solution into the liquid chromatograph and record the chromatogram. Separately, take an appropriate amount of tixotroxin reference substance and prepare a reference solution containing 10 μg per 1 ml with 0.03% acetic acid solution, and determine in the same method. Calculate the content of tixotroxin in the nanoparticles by the external standard method based on the peak area.

[0115] Method for determination of related substances: Accurately measure 1.0 ml of the test solution under the assay item, place it in a 100 ml volumetric flask, dilute to the mark with water, shake well, and use it as the control solution. According to the chromatographic conditions under the assay item, inject 50 μl of the control solution into the liquid chromatograph, adjust the detection sensitivity to make the peak height of the main component peak about 10% of the full scale; then inject 50 μl of the test solution and the reference solution into the liquid chromatograph respectively, and record the chromatogram until 2 times the retention time of the main component peak. The sum of the peak areas of each impurity peak in the chromatogram of the test solution shall not be greater than 3 times (3.0%) of the peak area of the main peak in the control solution.

[0116] Examples 1 - 4: Double-layer shell nanoparticles Feeding prescription table: See Table 1.

[0117] Table 1: Feeding prescription table for Examples 1 - 4 Preparation method: (S1) Preparation of primary emulsion (W / O): Dissolve the active ingredient with an aqueous solution containing a cosolvent to obtain Solution 1, and the concentration of the cosolvent in the aqueous solution containing the cosolvent is 0.1 wt%; then add Solution 1 to dichloromethane containing PEG-PLGA (50k), and in the dichloromethane containing PEG-PLGA (50k), add 100 mg of PEG-PLGA (50k) for every 1 ml of dichloromethane; homogenize at 10000 rpm for 5 min to form a primary emulsion (W / O); (S2) Preparation of multiple emulsion (W / O / W): Add the above primary emulsion (W / O) to an aqueous solution containing stabilizer A, and the content of stabilizer A in the aqueous solution containing stabilizer A is 1 wt%; homogenize at 10000 rpm for 5 min to form a multiple emulsion (W / O / W), add water and stir for 24 h to solidify, where 2 ml of water is added for every 1 ml of the multiple emulsion (W / O / W); centrifuge at 10000 rpm for 15 min to obtain solid A, wash the obtained solid A with water, and centrifuge at 10000 rpm for 15 min to obtain solid B, (S3)Formation of the second polymer matrix layer: The solid B obtained in step (S2) was added to dichloromethane containing PEG-PLGA (100k). In the dichloromethane containing PEG-PLGA (100k), 200 mg of PEG-PLGA (100k) was added per 1 ml of dichloromethane; homogenized at a high speed of 7000 rpm for 5 min to obtain emulsion 2; (S4)Formation of the double-layer shell nanoparticles: The emulsion 2 obtained in step (S3) was added to an aqueous solution containing stabilizer B. The content of stabilizer B in the aqueous solution containing stabilizer B was 1 wt%; homogenized at a high speed of 7000 rpm for 5 min to obtain emulsion 3, and stirred with water for 24 h for curing, where 2 ml of water was added per 1 ml of the emulsion 3; centrifuged at 10000 rpm for 15 min to obtain solid C, the obtained solid C was washed with water, centrifuged at 10000 rpm for 15 min to obtain solid D, and the obtained solid D was freeze-dried to obtain the double-layer shell nanoparticles.

[0118] The particle size, PDI and encapsulation efficiency of the obtained double-layer nanoparticles were detected, and the results are shown in Table 2.

[0119] Table 2: Results of particle size, PDI and encapsulation efficiency Examples 5 to 7: Investigation of the weight portion of PEG-PLGA (50k) Example 5: The difference from Example 4 was that by adjusting the dosage of PEG-PLGA (50k) in step (S1) to 500 weight portions, and the volume of dichloromethane in the dichloromethane containing PEG-PLGA (50k) in step (S1) remained unchanged, the ratio of dichloromethane to PEG-PLGA (50k) in the dichloromethane containing PEG-PLGA (50k) was adjusted to 25 mg of PEG-PLGA (50k) per 1 ml of dichloromethane, and the other conditions were the same as in Example 4.

[0120] Example 6: The difference from Example 4 was that by adjusting the dosage of PEG-PLGA (50k) in step (S1) to 1500 weight portions, and the volume of dichloromethane in the dichloromethane containing PEG-PLGA (50k) in step (S1) remained unchanged, the ratio of dichloromethane to PEG-PLGA (50k) in the dichloromethane containing PEG-PLGA (50k) was adjusted to 75 mg of PEG-PLGA (50k) per 1 ml of dichloromethane, and the other conditions were the same as in Example 4.

[0121] Example 7: The difference from Example 4 is that the dosage of PEG-PLGA (50k) in step (S1) is adjusted to 3,000 parts by weight, and the volume of dichloromethane in the dichloromethane containing PEG-PLGA (50k) in step (S1) remains unchanged, so that the ratio of dichloromethane to PEG-PLGA (50k) in the dichloromethane containing PEG-PLGA (50k) is adjusted to 150 mg of PEG-PLGA (50k) added per 1 ml of dichloromethane, and the remaining conditions are the same as in Example 4.

[0122] The particle size, PDI and encapsulation efficiency of the obtained double-layer nanoparticles were detected, and the results are shown in Table 3.

[0123] Table 3: Results of particle size, PDI and encapsulation efficiency Examples 8 to 10: Investigation of stabilizer A concentration Example 8: The difference from Example 2 is that by adjusting the feeding amount of stabilizer A (the amount of water in the aqueous solution containing stabilizer A remains unchanged), the content of stabilizer A in the aqueous solution containing stabilizer A in step (S2) is adjusted to 0.5 wt%, and the remaining conditions are the same as in Example 2.

[0124] Example 9: The difference from Example 2 is that by adjusting the feeding amount of stabilizer A (the amount of water in the aqueous solution containing stabilizer A remains unchanged), the content of stabilizer A in the aqueous solution containing stabilizer A in step (S2) is adjusted to 1.5 wt%, and the remaining conditions are the same as in Example 2.

[0125] Example 10: The difference from Example 2 is that by adjusting the feeding amount of stabilizer A (the amount of water in the aqueous solution containing stabilizer A remains unchanged), the content of stabilizer A in the aqueous solution containing stabilizer A in step (S2) is adjusted to 2 wt%, and the remaining conditions are the same as in Example 2.

[0126] The particle size, PDI and encapsulation efficiency of the obtained double-layer nanoparticles were detected, and the results are shown in Table 4.

[0127] Table 4: Results of particle size, PDI and encapsulation efficiency Examples 11 to 12: Investigation of parts by weight of PEG-PLGA (100k) Example 11: The difference from Example 3 is that the dosage of PEG-PLGA (100k) in step (S3) is adjusted to 1000 parts by weight, and the volume of dichloromethane in the dichloromethane containing PEG-PLGA (100k) in step (S3) remains unchanged, so that the ratio of dichloromethane to PEG-PLGA (50k) in the dichloromethane containing PEG-PLGA (100k) is adjusted to 50 mg of PEG-PLGA (100k) added per 1 ml of dichloromethane, and the remaining conditions are the same as those in Example 3.

[0128] Example 12: The difference from Example 3 is that the dosage of PEG-PLGA (100k) in step (S3) is adjusted to 3000 parts by weight, and the volume of dichloromethane in the dichloromethane containing PEG-PLGA (100k) in step (S3) remains unchanged, so that the ratio of dichloromethane to PEG-PLGA (50k) in the dichloromethane containing PEG-PLGA (100k) is adjusted to 150 mg of PEG-PLGA (100k) added per 1 ml of dichloromethane, and the remaining conditions are the same as those in Example 3.

[0129] The particle size, PDI and encapsulation efficiency of the obtained double-layer nanoparticles were detected, and the results are shown in Table 5.

[0130] Table 5: Results of particle size, PDI and encapsulation efficiency Comparative Example 1: Single-layer shell nanoparticles The difference from Example 4 is that operations in steps (S3) and (S4) are not carried out, but the double emulsion (W / O / W) obtained in step (S2) is added with water and stirred for 24 h for curing, where 2 ml of water is added per 1 ml of the double emulsion (W / O / W); centrifuged at 10000 rpm for 15 min to obtain solid A, the obtained solid A is washed with water, centrifuged at 10000 rpm for 15 min to obtain solid B, and the obtained solid B is freeze-dried to obtain single-layer shell nanoparticles.

[0131] The particle size, PDI and encapsulation efficiency of the obtained single-layer nanoparticles were detected, and the results are shown in Table 6.

[0132] Table 6: Results of particle size, PDI and encapsulation efficiency Comparative Examples 2 to 4: Investigation of the type of the innermost shell polymer matrix Comparative Example 2: The difference from Example 4 is that the polymer matrix PEG-PLGA (50k) of the innermost shell is replaced with DSPE-PEG (50k), and the rest is the same as in Example 4.

[0133] Comparative Example 3: The difference from Example 4 is that the polymer matrix PEG-PLGA (50k) of the innermost shell is replaced with DPPE-PEG (50k), and the rest is the same as Example 4.

[0134] Comparative Example 4: The difference from Example 4 is that the polymer matrix PEG-PLGA (50k) of the innermost shell is replaced with DMPE-PEG (50k), and the rest is the same as Example 4.

[0135] The particle size, PDI and encapsulation efficiency of the obtained double-layer nanoparticles were measured, and the results are shown in Table 7.

[0136] Table 7: Results of particle size, PDI and encapsulation efficiency Conclusion: Compared with Example 4, the nanoparticles prepared with the above polymer types have larger particle sizes and lower encapsulation efficiencies, and are not suitable for preparing the inner shell of double-layer nanoparticles. Therefore, the polymer PEG-PLGA (50k) was selected for preparing the inner shell of double-layer nanoparticles.

[0137] Comparative Examples 5 to 7: Investigation of the weight-average molecular weight of the polymer matrix of the innermost shell Comparative Example 5: The difference from Example 4 is that the weight-average molecular weight of the polymer matrix of the innermost shell is adjusted to 10 kDa, and the rest is the same as Example 4.

[0138] Comparative Example 6: The difference from Example 4 is that the weight-average molecular weight of the polymer matrix of the innermost shell is adjusted to 30 kDa, and the rest is the same as Example 4.

[0139] Comparative Example 7: The difference from Example 4 is that the weight-average molecular weight of the polymer matrix of the innermost shell is adjusted to 100 kDa, and the rest is the same as Example 4.

[0140] The particle size, PDI and encapsulation efficiency of the obtained double-layer nanoparticles were measured, and the results are shown in Table 8.

[0141] Table 8: Results of particle size, PDI and encapsulation efficiency Conclusion: The particle size increases with the increase in the molecular weight of the polymer. When the molecular weight of the polymer increases, the viscosity of the organic phase increases. Therefore, during the emulsification process, less organic phase is distributed in the aqueous phase, resulting in larger nanoparticles. When using PEG-PLGA (50k) to prepare nanoparticles, the encapsulation efficiency is the highest. Therefore, PEG-PLGA (50k) can be used as the polymer in the subsequent preparation process.

[0142] Comparative Examples 8 to 9: Investigation of the types of polymer matrices of the outer shell Comparative Example 8: The difference from Example 4 is that the polymer matrix PEG-PLGA (100k) of the outer shell is replaced with chitosan (100k), and the rest is the same as Example 4.

[0143] Comparative Example 9: The difference from Example 4 is that the polymer matrix PEG-PLGA (100k) of the outer shell is replaced with PLGA (100k), and the rest is the same as Example 4.

[0144] The particle size, PDI and encapsulation efficiency of the obtained double-layer nanoparticles were measured, and the results are shown in Table 9.

[0145] Table 9: Results of particle size, PDI and encapsulation efficiency Conclusion: The double-layer nanoparticles prepared with the above polymers have larger particle sizes and lower encapsulation efficiencies compared to PEG-PLGA (100k), and are not suitable for preparing the outer shell of double-layer nanoparticles. Therefore, the polymer PEG-PLGA (100k) is selected for preparing the outer shell of double-layer nanoparticles.

[0146] Comparative Examples 10 - 12: Investigation of the weight-average molecular weight of the polymer matrix of the outer shell Comparative Example 10: The difference from Example 4 is that the weight-average molecular weight of the polymer matrix of the outer shell is adjusted to 10 kDa, and the rest is the same as Example 4.

[0147] Comparative Example 11: The difference from Example 4 is that the weight-average molecular weight of the polymer matrix of the outer shell is adjusted to 30 kDa, and the rest is the same as Example 4.

[0148] Comparative Example 12: The difference from Example 4 is that the weight-average molecular weight of the polymer matrix of the outer shell is adjusted to 50 kDa, and the rest is the same as Example 4.

[0149] The particle size, PDI and encapsulation efficiency of the obtained double-layer nanoparticles were measured, and the results are shown in Table 10.

[0150] Table 10: Results of particle size, PDI and encapsulation efficiency Conclusion: Nanoparticle shells formed by PEG-PLGA with a larger molecular weight are thicker and the internal structure is denser, which can encapsulate drugs more effectively. It can also reduce the drug release rate and reduce the burst release phenomenon, which is consistent with the results of this experiment. Therefore, PEG-PLGA (100k) can be used as the outer polymer type for subsequent double-layer nanoparticles.

[0151] Comparative Examples 13 - 14: Investigation of the type of stabilizer Comparative Example 13: The difference from Example 4 is that the type of stabilizer A in step (S2) is replaced with polyethylene glycol succinate of vitamin E, and the other conditions are the same as those in Example 4.

[0152] Comparative Example 14: The difference from Example 4 is that the type of stabilizer A in step (S2) is replaced with Tween 80, and the other conditions are the same as those in Example 4.

[0153] The particle size, PDI and encapsulation efficiency of the obtained double-layer nanoparticles were detected, and the results are shown in Table 11.

[0154] Table 11: Results of particle size, PDI and encapsulation efficiency Conclusion: When using the surfactant PVA as a stabilizer, compared with the two surfactants TPGS and TWEEN 80, the encapsulation efficiency of its nanoparticles is the highest. As a non-ionic surfactant, PVA can form relatively small particles with a uniform size distribution and has excellent stability characteristics for nanoparticle emulsions.

[0155] Comparative Examples 15 - 17: Investigation of co-solvents Comparative Example 15: The difference from Example 4 is that citric acid is replaced with malic acid, and the other conditions are the same as those in Example 4.

[0156] Comparative Example 16: The difference from Example 4 is that citric acid is replaced with tartaric acid, and the other conditions are the same as those in Example 4.

[0157] Comparative Example 17: The difference from Example 4 is that citric acid is replaced with acetic acid, and the other conditions are the same as those in Example 4.

[0158] Investigation Example 1: Stability investigation The nanoparticles obtained in the above Examples and Comparative Examples were respectively placed at 25°C and 60% RH for 12 months, and samples were taken at the 0th, 6th, and 12th months to detect the content, particle size, encapsulation efficiency, related substances, etc. The results are shown in Table 12.

[0159] Table 12: Results of stability investigation Note: " / " indicates that the detection was not carried out. The acceptable standard for total impurities is ≤3.0%.

[0160] Conclusion: (1) For the type of the innermost shell polymer matrix, compared with using other polymer matrices (such as DSPE-PEG(50k), DPPE-PEG(50k) or DMPE-PEG(50k) in Comparative Example 2 to Comparative Example 4), using PEG-PLGA(50k) provided by the present invention is more conducive to reducing the particle size of the obtained nanoparticles, more conducive to improving the encapsulation efficiency of the obtained nanoparticles, more conducive to improving the stability of the content of tixotropium in the obtained nanoparticles, the stability of the encapsulation efficiency and the stability of related substances, and has unexpected technical effects.

[0161] (2) For the weight-average molecular weight of the innermost shell polymer matrix, compared with using other weight-average molecular weights (such as 10 kDa to 30 kDa in Comparative Example 5 to Comparative Example 6, or 100 kDa in Comparative Example 7), using the weight-average molecular weight provided by the present invention (such as 50 kDa) is more conducive to improving the encapsulation efficiency of the obtained nanoparticles, more conducive to improving the stability of the content of tixotropium in the obtained nanoparticles, the stability of the encapsulation efficiency and the stability of related substances, and has unexpected technical effects.

[0162] (3) For the type of the outer shell polymer matrix, compared with using other polymer matrices (such as chitosan(100k) or PLGA(100k) in Comparative Example 8 to Comparative Example 9), using PEG-PLGA(100k) provided by the present invention is more conducive to reducing the particle size of the obtained nanoparticles, more conducive to improving the encapsulation efficiency of the obtained nanoparticles, more conducive to improving the stability of the content of tixotropium in the obtained nanoparticles, the stability of the encapsulation efficiency and the stability of related substances, and has unexpected technical effects.

[0163] (4) For the weight-average molecular weight of the outer shell polymer matrix, compared with using other weight-average molecular weights (such as 10 kDa to 50 kDa in Comparative Example 10 - Comparative Example 12), using the weight-average molecular weight provided by the present invention (such as 100 kDa) is more conducive to improving the stability of the content of tixotropium in the obtained nanoparticles, the stability of the encapsulation efficiency and the stability of related substances, and has unexpected technical effects.

[0164] (5) For the type of the stabilizer, compared with using other stabilizers (such as polyethylene glycol vitamin E succinate or Tween 80 in Comparative Example 13 to Comparative Example 14), using PVA1788 provided by the present invention is more conducive to improving the encapsulation efficiency of the obtained nanoparticles, more conducive to improving the stability of the content of tixotropium in the obtained nanoparticles, the stability of the encapsulation efficiency and the stability of related substances, and has unexpected technical effects.

[0165] (6) Regarding the type of cosolvent, compared with the use of other cosolvents (such as malic acid, tartaric acid or acetic acid in Comparative Examples 15 to 17), the use of citric acid provided by the present invention is more conducive to improving the content stability and related substance stability of the obtained nanoparticles, and has unexpected technical effects.

[0166] Investigation Example 2: Investigation of in vitro release Accurately weigh 100 mg of the nanoparticles to be tested and place them in a 50 mL centrifuge tube. Add 5 mL of water and place it in a constant temperature magnetic stirrer at 37 °C with a stirring rate of 100 rpm / min. Take out the centrifuge tube at 0.5 h, 2 h, 6 h, 12 h, and 24 h after sample loading, place it in a centrifuge at 13000 rpm for 15 min to precipitate all the nanoparticles, carefully suck out 1 mL of the release medium and add 1 mL of PBS buffer, then continue stirring. Take the supernatant, filter it through a 0.22 µm aqueous membrane and inject the sample. The results are shown in Table 13.

[0167] Table 13: Results of in vitro release investigation Conclusion: (1) Compared with the nanoparticles with a single-layer shell, the present invention uses a double-layer shell to encapsulate the active ingredient, which greatly improves the sustained-release effect of the obtained nanoparticles and has unexpected technical effects.

[0168] The method of the present invention has been described through preferred embodiments. It is obvious that relevant personnel can make changes or appropriate modifications and combinations to the methods and applications described herein within the content, spirit and scope of the present invention to implement and apply the technology of the present invention. Those skilled in the art can draw on the content of this article and appropriately improve the process parameters to achieve it. It should be particularly noted that all similar substitutions and modifications are obvious to those skilled in the art, and they are all regarded as included in the present invention.

Claims

1. A core-shell nanoparticle, characterized in that: include: core and shell; The core comprises an active ingredient and a solubility aid; the active ingredient comprises tetrodoxin or a pharmaceutically acceptable salt thereof; The shell comprises at least two layers of nanoparticle shells; each layer of the nanoparticle shell comprises a polymer matrix layer and a stabilizer layer; The polymer matrix in the polymer matrix layer includes poly(lactic-co-glycolic acid)-poly(ethylene glycol).

2. The nanoparticles according to claim 1, wherein the cosolvent comprises at least one of citric acid, acetic acid, malic acid or tartaric acid, preferably citric acid; and / or The active ingredient is encapsulated in the innermost nanoparticle shell; and / or In each layer of the nanoparticle shell, there are a polymer matrix layer and a stabilizer layer from the inside to the outside; and / or In each layer of the nanoparticle shell, the stabilizer in the stabilizer layer independently includes at least one of polyvinyl alcohol, vitamin E polyethylene glycol succinate or Tween 80, preferably polyvinyl alcohol.

3. The nanoparticle according to any one of claims 1 to 2, wherein the weight average molecular weight of the poly(lactic acid-co-glycolic acid)-poly(ethylene glycol) in the innermost nanoparticle shell of the nanoparticle is 10 kDa-100 kDa, preferably 50 kDa; and / or The weight average molecular weight of the poly(lactic acid-co-glycolic acid)-poly(ethylene glycol) of the other nanoparticle shells outside the innermost nanoparticle shell in the nanoparticles is 10 kDa-100 kDa, preferably 100 kDa; and / or The polyvinyl alcohol is 1788 type.

4. The nanoparticle according to any one of claims 1 to 3, wherein the mass ratio of the active ingredient to the polymer matrix in the innermost nanoparticle shell of the nanoparticle is 1:100-1:1000 or 1:100-1:400 or 1:200-1:400; and / or The mass ratio of the active ingredient to the cosolvent is 1:0.3-1:3 or 1:1; and / or The mass ratio of the active ingredient to the polymer matrix in other nanoparticle shells outside the innermost nanoparticle shell of the nanoparticle is 1:200-1:2000 or 1:200-1:800 or 1:400-1:800; and / or In the innermost nanoparticle shell of the nanoparticle, the mass ratio of the polymer matrix to the stabilizer is 1:0.25-1:5 or 1:0.5-1:2.5 or 1:1-1:2.5; and / or In other nanoparticle shells outside the innermost nanoparticle shell of the nanoparticle, the mass ratio of the polymer matrix to the stabilizer is greater than 0:25 and less than or equal to 200:25, or 20:25 to 40:25; and / or The nanoparticles include two nanoparticle shells.

5. The nanoparticle according to any one of claims 1 to 4, wherein the proportions of the components of the nanoparticle are selected from the following groups: Group (1): The mass ratio of the active ingredient to the cosolvent is 1:1; The mass ratio of the active ingredient to the polymer matrix in the innermost nanoparticle shell of the nanoparticle is 1:100-1:2000; In the innermost nanoparticle shell of the nanoparticle, the mass ratio of the polymer matrix to the stabilizer is 1:0.5 to 1:2.5; The mass ratio of the active ingredient to the polymer matrix in the other nanoparticle shells outside the innermost nanoparticle shell of the nanoparticle is 1:200-1:800; In other nanoparticle shells outside the innermost nanoparticle shell of the nanoparticle, the mass ratio of the polymer matrix to the stabilizer is 20:25 to 40:25; Group (2): The mass ratio of the active ingredient to the cosolvent is 1:1; The mass ratio of the active ingredient to the polymer matrix in the innermost nanoparticle shell of the nanoparticle is 1:200-1:400; In the innermost nanoparticle shell of the nanoparticle, the mass ratio of the polymer matrix to the stabilizer is 1:1 to 1:2.5; The mass ratio of the active ingredient to the polymer matrix in the other nanoparticle shells outside the innermost nanoparticle shell of the nanoparticle is 1:400-1:800; In other nanoparticle shells outside the innermost nanoparticle shell of the nanoparticle, the mass ratio of the polymer matrix to the stabilizer is 20:25 to 40:25; Group (3): The mass ratio of the active ingredient to the cosolvent is 1:1; The mass ratio of the active ingredient to the polymer matrix in the innermost nanoparticle shell of the nanoparticle is 1:200; In the innermost nanoparticle shell of the nanoparticle, the mass ratio of the polymer matrix to the stabilizer is 1:2.5; The mass ratio of the active ingredient to the polymer matrix in the other nanoparticle shells outside the innermost nanoparticle shell of the nanoparticle is 1:800; In the other nanoparticle shells outside the innermost nanoparticle shell of the nanoparticle, the mass ratio of the polymer matrix to the stabilizer is 40:25; Group (4): The mass ratio of the active ingredient to the cosolvent is 1:1; The mass ratio of the active ingredient to the polymer matrix in the innermost nanoparticle shell of the nanoparticle is 1:200; In the innermost nanoparticle shell of the nanoparticle, the mass ratio of the polymer matrix to the stabilizer is 1:2; The mass ratio of the active ingredient to the polymer matrix in the other nanoparticle shells outside the innermost nanoparticle shell of the nanoparticle is 1:800; In the other nanoparticle shells outside the innermost nanoparticle shell of the nanoparticle, the mass ratio of the polymer matrix to the stabilizer is 40:25; Group (5): The mass ratio of the active ingredient to the cosolvent is 1:1; The mass ratio of the active ingredient to the polymer matrix in the innermost nanoparticle shell of the nanoparticle is 1:400; In the innermost nanoparticle shell of the nanoparticle, the mass ratio of the polymer matrix to the stabilizer is 1: 1.25; The mass ratio of the active ingredient to the polymer matrix in the other nanoparticle shells outside the innermost nanoparticle shell of the nanoparticle is 1:400; In the other nanoparticle shells outside the innermost nanoparticle shell of the nanoparticle, the mass ratio of the polymer matrix to the stabilizer is 20:25; Group (6): The mass ratio of the active ingredient to the cosolvent is 1:1; The mass ratio of the active ingredient to the polymer matrix in the innermost nanoparticle shell of the nanoparticle is 1:400; In the innermost nanoparticle shell of the nanoparticle, the mass ratio of the polymer matrix to the stabilizer is 1:2.5; The mass ratio of the active ingredient to the polymer matrix in the other nanoparticle shells outside the innermost nanoparticle shell of the nanoparticle is 1:800; In the other nanoparticle shells outside the innermost nanoparticle shell of the nanoparticle, the mass ratio of the polymer matrix to the stabilizer is 40:

25.

6. A method for preparing the nanoparticles according to any one of claims 1 to 5, characterized in that: The following steps are involved: (S1) Preparation of colostrum (W / O): dissolving the active ingredient in an aqueous solution containing a cosolvent to obtain a solution 1, and then adding the solution 1 to an organic solvent containing a polymer matrix, and homogenizing at a high speed to form colostrum (W / O); (S2) Preparation of a double emulsion (W / O / W): adding the colostrum (W / O) to an aqueous solution containing a stabilizer, homogenizing at high speed to form a double emulsion (W / O / W), adding water and stirring to solidify, centrifuging to obtain a solid A, washing the obtained solid A, centrifuging to obtain a solid B; (S3) Forming a second polymer matrix layer: adding the solid B obtained in step (S2) to an organic solvent containing a polymer matrix, and homogenizing at high speed to obtain an emulsion 2; (S4) Formation of double-shell nanoparticles: adding the emulsion 2 obtained in step (S3) to an aqueous solution containing a stabilizer, homogenizing at high speed to obtain emulsion 3, adding water and stirring to solidify, centrifuging to obtain solid C, washing the obtained solid C, centrifuging to obtain solid D, and freeze-drying the solid D to obtain double-shell nanoparticles.

7. The method according to claim 6, wherein the organic solvent in step (S1) or step (S3) is independently selected from dichloromethane; and / or The concentration of the co-solvent in the aqueous solution containing the co-solvent in step (S1) is 0.1wt%~1wt% or 0.1wt%; and / or In the organic solvent containing the polymer matrix in step (S1), 25 mg-300 mg or 100 mg of the polymer matrix is ​​added per 1 ml of the organic solvent; and / or In the aqueous solution containing the stabilizer in step (S2), the concentration of the stabilizer is 0.5wt%-2wt% or 1wt%; and / or In the water-adding, stirring and solidifying operation of step (S2), the volume ratio of the emulsion (W / O / W) to water is greater than 0:2 and less than or equal to 1:2, or is 1:2; and / or The time of adding water, stirring and curing in step (S2) is 0-24 h, or 24 h; and / or The speed of any centrifugation in step (S2) is independently selected from 5000 rpm-15000 rpm or 10000 rpm; and / or The centrifugation time of any centrifugation in step (S2) is independently selected from 5 min -30 min, or 15 min; and / or The cleaning in step (S2) is cleaning with water; and / or In the organic solvent of the polymer matrix in step (S3), 25 mg-300 mg or 100 mg of the polymer matrix is ​​added per 1 ml of the organic solvent; and / or In the aqueous solution containing the stabilizer in step (S4), the concentration of the stabilizer is 0.5 wt%-2 wt% or 1 wt%; and / or The weight average molecular weight of the polymer matrix in step (S1) is 10 kDa-100 kDa or 50 kDa; and / or The weight average molecular weight of the polymer matrix in step (S3) is 10 kDa-100 kDa or 100 kDa; and / or The rotation speed of the high-speed homogenization in step (S1) is 2000 rpm-10000 rpm or 10000 rpm; and / or The time of high-speed homogenization in step (S1) is 2 min-5 min or 5 min; and / or The speed of the high-speed homogenization in step (S2) is 2000 rpm-10000 rpm or 10000 rpm; and / or The time of high-speed homogenization in step (S2) is 2 min-5 min or 5 min; and / or The speed of the high-speed homogenization in step (S3) is 2000 rpm-10000 rpm or 7000 rpm; and / or The time of high-speed homogenization in step (S3) is 2 min-5 min or 5 min; and / or The speed of the high-speed homogenization in step (S4) is 2000 rpm-10000 rpm or 7000 rpm; and / or The time of high-speed homogenization in step (S4) is 2 min-5 min or 5 min; and / or In the water-adding, stirring and solidifying operation of the step (S4), the volume ratio of the emulsion 3 to the water is greater than 0:2 and less than or equal to 1:2, or is 1:2; and / or The time of adding water, stirring and curing in step (S4) is 0-24 h, or 24 h; and / or The centrifugal speed of any centrifugation in step (S4) is independently selected from 5000 rpm-15000 rpm or 10000 rpm; and / or The centrifugation time of any centrifugation in step (S4) is independently selected from 5 min -30 min, or 15 min; and / or The cleaning in step (S4) is performed by washing with water.

8. A pharmaceutical composition, characterized in that The invention comprises the nanoparticles according to any one of claims 1 to 5 or the nanoparticles prepared by the method according to any one of claims 6 to 7, or comprises the nanoparticles according to any one of claims 1 to 5 or the nanoparticles prepared by the method according to any one of claims 6 to 7 and a pharmaceutically acceptable excipient or carrier.

9. A pharmaceutical preparation, characterized in that The nanoparticles according to any one of claims 1 to 5 or the nanoparticles prepared by the method according to any one of claims 6 to 7 or the pharmaceutical composition according to claim 8; Optionally, the pharmaceutical preparation is an injection; Optionally, the pharmaceutical preparation is a sustained-release injection or a controlled-release injection.

10. Use of the nanoparticles according to any one of claims 1 to 5, the nanoparticles prepared by the method according to any one of claims 6 to 7, the pharmaceutical composition according to claim 8 or the pharmaceutical preparation according to claim 9 in the preparation of drugs for analgesia, drug addiction treatment and / or antiarrhythmia.

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