WO2021057007A1

WO2021057007A1 - Rapamycin nanoscale sustained-release agent and preparation method thereof

Info

Publication number: WO2021057007A1
Application number: PCT/CN2020/082852
Authority: WO
Inventors: 严鹏科
Original assignee: 严鹏科
Priority date: 2019-09-26
Filing date: 2020-04-01
Publication date: 2021-04-01
Also published as: CN110623925A; CN110623925B

Abstract

Disclosed is a rapamycin nanoscale sustained-release agent prepared from the following raw materials in parts by weight: 1 part of rapamycin, 0.5-20 parts of soluble polymer carrier, 40-200 parts of organic solvent, and 400-20000 parts of aqueous phase. The rapamycin nanoscale sustained-release agent has a structure of nanomicelle and a particle size of 10-200 nm. The rapamycin nanoscale sustained-release agent presents low risk to blood vessels. The half-life of the rapamycin nanoscale sustained-release agent in the blood can be up to 50 hours or more. The rapamycin nanoscale sustained-release agent can directly reach the tumor-affected area. With continuous medication, the tumor regression rate can reach 50%.

Description

Rapamycin nano sustained-release agent and preparation method thereof

Technical field

The invention relates to the technical field of rapamycin preparations, in particular to a rapamycin nano sustained-release agent and a preparation method thereof.

Background technique

Tumor Cancer has become the number one killer that endangers human health. Although there are endless methods to treat tumors, the living conditions of most patients have not been greatly improved. Among the various treatment methods for tumors, chemotherapy is still the most commonly used option. Although chemotherapy drugs are widely used, their therapeutic effects on solid tumors are not exact. The fundamental problem is that traditional chemotherapeutic drugs cannot achieve effective therapeutic concentrations at the tumor site or cannot maintain sufficient time of action. Moreover, traditional chemotherapeutic drugs can kill normal cells indiscriminately, resulting in a variety of toxic side effects. The effect of chemotherapeutic drugs depends not only on the sensitivity of the drug, but also on the time of action of the drug at the tumor site and the accumulation of the drug at the tumor site. Therefore, the local application of chemotherapeutic drugs, especially local sustained release, has become a hot and difficult point in current tumor chemotherapy research.

Rapamycin was discovered in the soil of Easter Island in Chile in 1975. It is a hydrophobic macrolide immunosuppressant produced by streptomyces hygroscopicus. It has antifungal activity and is a white crystal with a relative molecular mass of 914.2. It is easily soluble in organic solvents such as formaldehyde, ethanol, acetone, chloroform, and almost insoluble in water.

Rapamycin is a powerful immunosuppressant with low toxicity. It inhibits the cell cycle G ₀ and G ₁ phases by combining with the corresponding immunophilin RMBP, and blocks G _{1 from} entering the S phase. It is widely used in transplantation surgery. in. In addition to immunosuppressive effects, rapamycin also has anti-tumor effects, which can inhibit the growth of tumor cells such as kidney cancer, lymphoma, lung cancer, liver cancer, breast cancer, neuroendocrine cancer and gastric cancer in a concentration-dependent manner. Since 2007, two derivatives of rapamycin, tamsulolimus and everolimus, have been developed for the treatment of cancer. The research and application of rapamycin in tumor treatment has been increasing. It is used alone or in combination in vitro , Both show significant anti-tumor effects in vivo. RAPA inhibits mammalian target of rapamycin (m TOR) receptors and affects various signal pathways of its transduction, thereby exerting various effects on tumors such as anti-angiogenesis, blocking cell cycle and promoting cell apoptosis. The process of proliferation, invasion and metastasis has an impact.

Nano-preparation has a high degree of dispersibility and a huge surface area, which is beneficial to increase the contact time and contact area of the drug and the biofilm of the absorption site, and increase the solubility of the drug; the nanoparticle can enter the cell through the endocytosis mechanism and transport the general drug across the membrane. The mechanism is different, so it may increase the permeability of the drug to the biofilm. Nano drug delivery system as an effective means to optimize drug efficacy has become a research hotspot in the fields of pharmacy and modern biomedicine.

Rapamycin is a hydrophobic drug and cannot be directly used for injection. It needs a certain organic solvent to dissolve it before injection, which is easy to cause adverse effects on the human body; the bioavailability of rapamycin in vivo is very low, and it is easy to cause failure to reach the diseased site It's already a problem.

technical problem

Currently, there are phospholipids or cholesterones on the market as the carrier of nano-released rapamycin. This type of carrier has a very high affinity with the human body, but due to the natural degradation rate of phospholipids or cholesterones in the blood Fast, so that the concentration of rapamycin at the site of action is low, and the targeting is insufficient.

Technical solutions

In order to overcome the shortcomings of the prior art, one of the objectives of the present invention is to provide a rapamycin that can effectively control the sustained release rate of rapamycin, increase its residence time in tumor tissues, and prolong its half-life in plasma. Nanometer slow release agent.

The second object of the present invention is to provide a preparation method of the rapamycin nano sustained-release agent.

One of the objectives of the present invention is achieved by adopting the following technical solutions:

A rapamycin nano sustained-release agent, which is made from the following raw materials in parts by weight: 1 part of rapamycin, 0.5-20 parts of soluble polymer carrier, 40-200 parts of organic solvent and 400-20000 Parts of the aqueous phase.

The nano slow-release agent forms a slow-release coating structure through the coating effect of a soluble high molecular polymer on rapamycin. At the same time, the coating structure is dispersed into nano-scale through the dispersibility of the organic solvent and the aqueous phase. Of coated particles.

Further, the soluble high molecular polymer carrier is polyethylene glycol-2000, polyethylene glycol-4000, polyethylene glycol-10000, polyethylene glycol-15000, PLGA, PEO, PVP, polypropylene, poly One or two or more of amino acid, polysorbate and polyoxyethylene ester fatty acid. Compared with the carrier of small molecular weight, the sustained-release agent formed by the polymer carrier has a relatively longer hydrophilic time, which can increase the sustained-release time of the injection formed by rapamycin in the body and prolong the half-life.

Further, the soluble high molecular polymer carrier is a methoxy polyethylene glycol block copolymer. When the soluble high-content polymer is formulated into a sustained-release agent, it can effectively form nanoparticles with a controllable sustained-release time.

Further, the soluble high molecular polymer carrier is mPEG-PLA with a molecular weight of 3000-20000. It is a block copolymer of methoxy polyethylene glycol and polylactic acid.

Further, the organic solvent is one or more of absolute ethanol, dichloromethane, acetone and methanol. The organic solvent needs to have better solubility for rapamycin and the soluble high molecular polymer carrier, and at the same time have better dispersibility in water, and be miscible with water. That is, the organic solvent is soluble in water.

Further, the aqueous phase liquid is one or two of distilled water, physiological saline, cell culture fluid, body fluid, tissue fluid, buffer, or glucose injection. The aqueous phase liquid provides a better dispersion medium for the organic phase. Through the dispersion of the hydrophilic soluble polymer carrier and the organic solvent, the dispersibility of rapamycin in the aqueous phase can be effectively improved to form Nano-level particles.

Further, the raw material also includes a freeze-dried protective agent. The lyoprotectant is one or more of lactose, glucose, mannitol or sucrose.

The second objective of the present invention is achieved by adopting the following technical solutions:

A preparation method of the above-mentioned rapamycin nano sustained-release agent includes the following steps:

1) Add rapamycin bulk drug and soluble polymer carrier to organic solvent to form organic phase;

2) Inhale the organic phase into the syringe, add 1-10 drops per minute to the aqueous phase, and stir for 30min-3h at room temperature;

3) Recover organic solvents under reduced pressure;

4) Centrifuge for 5-120min, take the supernatant, filter with 0.22-0.45μm membrane to obtain micellar solution;

5) Freeze-dry the micellar solution to obtain a rapamycin nano-sustained release agent.

That is, in this method, the rapamycin bulk drug and the soluble high molecular polymer carrier are dispersed by an organic solvent to form a uniformly dispersed organic phase. Then, by slowly releasing the organic phase into the aqueous phase, the difference in dissolution rate and the agitation of the aqueous phase digestion cause the rapamycin and the soluble high molecular polymer to form nano-scale micelles.

Further, in step 2), the stirring speed is 500-800 rpm; in step 4), the centrifugal speed is 4000-8000 rpm.

Further, in step 4), 5-10 g of lyoprotectant is added to every 100 mL of micellar solution.

Beneficial effect

The invention provides a rapamycin nano sustained-release agent, which finally forms a micelle-like nano particle structure in an aqueous phase through the coating effect of a soluble high molecular polymer and the solubilization and dispersion effect of an organic solvent. The micellar nanoparticle structure improves the sustained release of rapamycin in plasma, tissue fluid or further in the digestive tract, and its half-life is relatively controllable, allowing rapamycin to reach the affected area.

The rapamycin nano sustained-release agent provided by the present invention has a nano-micelle structure with a particle size of 10-200nm, a drug loading amount of 0.1-20%, and an encapsulation rate of over 80%. It has uniformity and Stable particle size distribution, stable encapsulation rate and drug loading, and low risk to blood vessels; the rapamycin nano-released agent has stable encapsulation rate and drug loading, and has a better tumor targeting effect;

The half-life of the rapamycin nano sustained-release agent in the blood can be as high as more than 50 hours, and it can reach the affected area of the tumor directly. With continuous medication, the tumor regression rate can reach 50%.

Description of the drawings

Figure 1 shows the appearance of the formulations of Examples 1-5;

Figure 2 is a characterization diagram of the rapamycin nano sustained-release agent of Example 4;

Fig. 3 is a graph of the in vitro anti-tumor test results of the rapamycin nano-sustained release agent of Example 4;

Figure 4 is a diagram showing the results of in vivo targeting of the rapamycin nano sustained-release agent of Example 4;

Fig. 5 is a diagram showing the anti-tumor effect of the rapamycin nano sustained-release agent of Example 4 in vivo.

The best mode of the present invention

In the following, the present invention will be further described with reference to the drawings and specific implementations. It should be noted that, provided that there is no conflict, the following embodiments or technical features can be arbitrarily combined to form new embodiments. .

That is, the application uses the coating effect of the soluble polymer carrier and the dispersion effect of the organic solvent to coat rapamycin in it to form micelles, which are slowly released into the aqueous phase to form nano-sized particles. The structure is filtered and lyophilized to obtain a rapamycin nano-sustained release agent that can be used for injection.

The soluble high molecular polymer carrier is preferably a methoxy polyethylene glycol block copolymer, that is, it has a methoxy end group that can have better compatibility with rapamycin. Preferably, it is an mPEG-PLA block copolymer, and the molecular weight is preferably 2000-20000 to form micelles, thereby forming nano-sustained-release agent particles with high encapsulation efficiency and an average particle size in the nm level. In the following specific embodiments, Examples 1-10 all use mPEG-PLA copolymers, wherein the molecular weight of mPEG is 2000, the molecular weight of PLA is 2000, and the total molecular weight is 4000.

The residence time of the rapamycin nano-released agent in the tumor is 24-48h; the half-life in the blood is more than 52h. The rapamycin nano-released agent has an injection amount of rapamycin based on the effective ingredient. Above 10µg/mL, the tumor tissue will disappear by at least 50% after continuous medication.

The preparation method of the rapamycin nano sustained-release agent includes the following steps:

3) Recover organic solvents under reduced pressure;

In the preparation process of the rapamycin nano-sustained release agent, through the mutual dissolution process of the organic phase and the aqueous phase, the micelles with small particle size are formed by physical force through the method of stirring and dispersing; the final method is centrifugation, The free rapamycin in the micellar solution is removed, thereby obtaining a rapamycin nano-release drug with low blood health risk.

The carrier of the rapamycin nano sustained-release agent can be degraded under natural physiological conditions, thereby being excreted from the body through metabolism, and will not produce irritation or foreign body reaction to the body.

The following are specific embodiments of the present invention. The raw materials, equipment, etc. used in the following embodiments can be obtained through purchase except for special restrictions.

Embodiments of the present invention

Example 1 :

A rapamycin nano sustained-release agent, made of the following components: 5 mg rapamycin, 10 mg mPEG-PLA block polymer, 1 mL acetone and 50 mL PBS buffer;

The preparation method includes the following steps:

1) Add rapamycin bulk drug and mPEG-PLA block polymer to acetone to form an organic phase;

2) Inhale the organic phase into the syringe, add 5 drops per minute to the stirring PBS buffer at 500 rpm, and stir at 600 rpm for 60 minutes at room temperature;

3) Recover organic solvents under reduced pressure at 40°C;

4) Centrifuge at 4000 r/min for 30 min, take the supernatant, filter and sterilize with a 0.22 µm pore size microporous membrane to obtain a micellar solution;

Example 2 :

A rapamycin nano sustained-release agent, made of the following components: 5 mg rapamycin, 20 mg mPEG-PLA block polymer, 1 mL acetone and 50 mL PBS buffer;

The preparation method includes the following steps:

3) Recover organic solvents under reduced pressure at 40°C;

4) Centrifuge at 5000 r/min for 30 min, take the supernatant, filter and sterilize with a 0.22 µm pore size microporous membrane to obtain a micellar solution;

Example 3 :

A rapamycin nano sustained-release agent, made of the following components: 5 mg rapamycin, 40 mg mPEG-PLA block polymer, 1 mL acetone and 50 mL PBS buffer;

The preparation method includes the following steps:

3) Recover organic solvents under reduced pressure at 40°C;

4) Centrifuge at 5000 r/min for 30 minutes, take the supernatant, filter and sterilize with a 0.22μm pore size microporous membrane to obtain a micellar solution;

Example 4 :

A rapamycin nano sustained-release agent, made of the following components: 5 mg rapamycin, 50 mg mPEG-PLA block polymer, 1 mL acetone and 50 mL PBS buffer;

The preparation method includes the following steps:

3) Recover organic solvents under reduced pressure at 40°C;

Example 5 :

A rapamycin nano sustained-release agent, made of the following components: 5 mg rapamycin, 60 mg mPEG-PLA block polymer, 1 mL acetone and 50 mL PBS buffer;

The preparation method includes the following steps:

2) Inhale the organic phase into the syringe, add 5 drops per minute to the stirring PBS buffer at 500 rpm, and stir at 600 rpm for 120 minutes at room temperature;

3) Recover organic solvents under reduced pressure at 40°C;

Example 6 :

A rapamycin nano sustained-release agent, made of the following components: 10 mg rapamycin, 90 mg mPEG-PLA block polymer, 1 mL acetone and 100 mL PBS buffer;

The preparation method includes the following steps:

2) Inhale the organic phase into the syringe, add 5 drops per minute to the stirring PBS buffer at 500 rpm, and stir at 800 rpm at room temperature for 120 minutes;

3) Recover organic solvents under reduced pressure at 40°C;

4) Centrifuge at 4000 r/min for 30 minutes, take the supernatant, filter and sterilize with a 0.22 µm pore size microporous filter membrane to obtain a micellar solution;

Example 7 :

A rapamycin nano sustained-release agent, which is made of the following components: 100 mg rapamycin, 900 mg mPEG-PLA block polymer, 7 mL acetone, 10 mL PBS buffer and 0.5 g lactose;

The preparation method includes the following steps:

3) Recover organic solvents under reduced pressure at 40°C;

5) Add lactose to the micellar solution, aseptically filter with a 0.22 µm pore filter membrane, freeze-dry, and obtain a rapamycin nano-sustained release agent.

Example 8 :

A rapamycin nano sustained-release agent, which is made of the following components: 150 mg rapamycin, 100 mg mPEG-PLA block polymer, 7 mL acetone, 100 mL PBS buffer and 5 g lactose;

The preparation method includes the following steps:

3) Recover organic solvents under reduced pressure at 40°C;

Example 9 :

A rapamycin nano sustained-release agent, made of the following components: 100 mg rapamycin, 1000 mg mPEG-PLA block polymer, 7 mL acetone, 100 mL PBS buffer and 5 g lactose;

The preparation method includes the following steps:

3) Recover organic solvents under reduced pressure at 40°C;

Example 10 :

A rapamycin nano sustained-release agent, which is made of the following components: 50 mg rapamycin, 500 mg mPEG-PLA block polymer, 7 mL acetone, 50 mL PBS buffer and 2.5 g lactose;

The preparation method includes the following steps:

3) Recover organic solvents under reduced pressure at 40°C;

Performance testing:

1. Preparation performance test

The appearance evaluation, average particle size, potential and encapsulation efficiency of the rapamycin nano-sustained release agent obtained in Examples 1-10 were measured;

Among them, the appearance evaluation criteria: to maintain the original volume, no collapse, no shrinkage, uniform color, no variegation, and fine texture; the appearance is shown in Figure 1, and from left to right are the preparations of Examples 1-5 . ...

Average particle size: Malvern laser particle size analyzer is used to measure the particle size and particle size distribution of nanoparticles. The principle is to use the characteristics of light scattering and light diffraction when the particles are irradiated by light, and the light scattering intensity and diffraction intensity are compared with the particles. The principle related to size and optical characteristics is used to determine the particle size.

As shown in Figure 2, Figure 2A is a mimic diagram of micelles of the rapamycin nano sustained-release agent of Example 4, in which the spherical shape is the active ingredient rapamycin, and the linear part is the mPEG-PLA block polymer; Figure 2B is a particle size distribution diagram of rapamycin nano sustained-release agent; Figure 2C is a TEM image of rapamycin nano sustained-release agent.

Potential: A Malvern laser particle size analyzer was used to measure the potential of the nanoparticles; Figure 2D is the Zeta potential diagram of the rapamycin nano-sustained release agent.

Encapsulation rate: The encapsulation rate is better than 80%.

Refer to the content determination method to determine the total content of the drug.

The drug content was determined by high performance liquid chromatography with methanol-acetonitrile-water (volume ratio 43:40:17) as mobile phase, flow rate of 1 mL/min, column temperature of 40 ℃, and detection wavelength of 278 nm.

The formula for calculating the encapsulation rate is: encapsulation rate=encapsulated drug amount/total content of main drug×100%

The results are shown in the following table:

Table 1 Changes in the encapsulation efficiency, drug loading and average particle size of nanoparticles

To	外观Exterior	再分散性Redispersibility	包封率%Encapsulation rate%	平均粒径(nm)Average particle size (nm)
实施例1Example 1	无皱缩，无塌陷No shrinkage, no collapse	良好good	9191	12.3712.37
实施例2Example 2	无皱缩，无塌陷No shrinkage, no collapse	良好good	8585	17.617.6
实施例3Example 3	无皱缩，无塌陷No shrinkage, no collapse	良好good	8787	18.518.5
实施例4Example 4	无皱缩，无塌陷No shrinkage, no collapse	良好good	8888	18.918.9
实施例5Example 5	无皱缩，无塌陷No shrinkage, no collapse	良好good	8686	18.618.6
实施例6Example 6	无皱缩，无塌陷No shrinkage, no collapse	良好good	8787	28.128.1
实施例7Example 7	无皱缩，无塌陷No shrinkage, no collapse	良好good	8585	25.325.3
实施例8Example 8	无皱缩，无塌陷No shrinkage, no collapse	良好good	8484	29.429.4
实施例9Example 9	无皱缩，无塌陷No shrinkage, no collapse	良好good	8787	31.331.3
实施例10Example 10	无皱缩，无塌陷No shrinkage, no collapse	良好good	8888	27.527.5

It can be seen from Table 1 that the encapsulation rate of the rapamycin nano sustained-release preparation obtained in this application is above 80%.

2. Preparation performance test

The MTT kit method and HCT116 cells were used for cytotoxicity experiments. HCT116 cells were seeded in a 96-well plate/5% CO ₂ ^{/37°C incubator at an inoculum of 1×10 4} cells/well for 24 hours, and the concentrations were given respectively. (Based on the effective ingredients of sodium rapamycin) 80 µg/mL, 40.00 µg/mL, 30.00 µg/mL, 20.00 µg/mL, 10.00 µg/mL, 5.00 µg/mL, 2.50 µg/mL, 1.25 µg/ After treatment with the rapamycin nano-release agent of Example 4 in mL, 0.65 µg/mL, 0.3125 µg/mL, and 0 µg/mL for 24h, 48h, and 72h, respectively, the rapamycin nano-release agent significantly inhibited HCT116 The growth of the cells is shown in Figure 3. The IC ₅₀ for 24 hours of administration is 10.29 µg/mL, the IC ₅₀ for 48 hours of administration is 3.92 µg/mL, and the IC ₅₀ for 72 hours of administration is 0.63 µg/mL.

3. In vivo tumor targeting effect of rapamycin nano sustained-release agent

Modeling of HCT116 solid tumor mice: Take 20 BALB/c nude mice, females, weighing 20 g, and inoculate 0.2 mL subcutaneously with the prepared HCT116 cell suspension, the number of cells is 5×10 ⁶ .

Group administration: After vaccination, they were randomly divided into five groups. The dosage of rapamycin nano sustained-release agent was 13.3μg, 40μg and 120μg groups, namely low, medium and high dose groups, and a rapamycin control group of 40μg and Normal saline control group. Four animals in each group were administered by tail vein injection with a 0.2 mL administration volume on the fifth day after vaccination, once every two days (approximately 56 hours), for 21 consecutive days.

Tumor volume was measured every other day after administration. 50 hours after the last administration, the mice were weighed, blood was taken, the mice were sacrificed, the liver was taken, the tumor was taken, the blood was taken, and the content of rapamycin in the liver and tumor was taken. The results are shown in Table 2 and Figure 4. Figure 4A is the content of rapamycin in the blood; Figure 4B is the content of rapamycin in the liver; Figure 4C is the content of rapamycin in the tumor;

Table 2 The content of rapamycin in blood, liver and tumor

To	对照组40µgControl group 40µg	试验组13.3µgTest group 13.3µg	试验组40µgTest group 40µg	试验组120µgTest group 120µg
血（ng/mL）Blood (ng/mL)	53.04±18.9653.04±18.96	97.97±10.497.97±10.4	114.01±2.61114.01±2.61	143.71±8.33143.71±8.33
肝(ng/mg)Liver (ng/mg)	0.042±0.000.042±0.00	0.083±0.010.083±0.01	0.084±0.000.084±0.00	0.138±0.040.138±0.04
肿瘤(ng/mg)Tumor (ng/mg)	////	0.11±0.050.11±0.05	0.37±0.060.37±0.06	0.38±0.050.38±0.05

It can be seen from Table 2 that, relative to the rapamycin control group, 50 hours after administration, the content of rapamycin in the nano-sustained-release preparation group of the first draft example 4 at the same dose was more than twice that of the control group, and There are still residues in the tumor.

The rapamycin in Example 4 was replaced with DiR liposomes to make a nano-released DiR liposome. As a control group, after the tail vein was injected into nude mice, live imaging was performed at different time points to observe the position of fluorescence . It was found that after 18 hours, the nanoparticles gathered at the tumor site. The results are shown in Figure 4, 4D and 4E. Figure 4D shows the targeting effect of the DiR liposome nano-release agent on tumor tissues at different time points; Figure 4E shows the fluorescence results of the internal organs and tumor tissues taken out by the DiR liposome nano-release agent for 24 hours.

It can be seen from Figs. 4D and 4E that the sustained-release agent of the present application can make the effective ingredients reach the tumor directly, has a long-lasting action time, and has a better targeting effect.

4. Anti-tumor effect of rapamycin nano-sustained release agent in vivo

HCT116 solid tumor mouse model: Take 20 BALB/c nude mice, female, weight (20) g, and subcutaneously inoculate 0.2 mL of the mouse with the prepared HCT116 cell suspension, the cell number is 5×10 ⁶ A.

Grouped administration: Randomly divided into five groups after inoculation. The rapamycin nano sustained-release agent in Example 4 is 13.3μg, 40μg, and 120μg groups, namely low, medium and high dose groups, and a rapamycin control group 40μg And the normal saline control group. Four animals in each group were administered by tail vein injection with a 0.2 mL administration volume on the fifth day after vaccination, once every two days (approximately 56 hours). The administration was continued for 21 days.

Tumor volume was measured every other day after administration. 50 hours after the last administration, the mice were weighed, the mice were sacrificed, the tumors were taken, the tumors were weighed, and the tumor inhibition rate of each group was calculated.

Figure 5A is the dosage regimen for nude mice; Figure 5B is the tumor volume after the administration of rapamycin nano-released agent; Figure 5C is the tumor weight after the administration of rapamycin nano-released agent. It can be seen from the above figure that, compared with the direct administration of ordinary rapamycin, the nano sustained-release agent of the present application can significantly inhibit the growth of tumors.

The foregoing embodiments are only preferred embodiments of the present invention, and cannot be used to limit the scope of protection of the present invention. Any insubstantial changes and substitutions made by those skilled in the art on the basis of the present invention belong to the present invention. The scope of protection required.

Claims

A rapamycin nano sustained-release agent, characterized in that it is made of the following raw materials in parts by weight: 1 part of rapamycin, 0.5-20 parts of soluble polymer carrier, and 40-200 parts of organic solvent And 400-20000 parts of water phase liquid.
The rapamycin nano sustained-release agent according to claim 1, wherein the soluble polymer carrier is polyethylene glycol-2000, polyethylene glycol-4000, polyethylene glycol-10000, One or more of polyethylene glycol-15000, PLGA, PEO, PVP, polypropylene, polyamino acid, polysorbate and polyoxyethylene ester fatty acid.
The rapamycin nano sustained-release agent according to claim 1, wherein the soluble polymer carrier is a methoxy polyethylene glycol block copolymer.
The rapamycin nano sustained-release agent according to claim 1, wherein the soluble high molecular polymer carrier is mPEG-PLA with a molecular weight of 2000-20000.
The rapamycin nano sustained-release agent according to claim 1, wherein the organic solvent is one or more of absolute ethanol, dichloromethane, acetone and methanol.
The rapamycin nano sustained-release agent according to claim 1, wherein the aqueous phase liquid is one or two of distilled water, physiological saline, cell culture fluid, body fluid, tissue fluid, buffer, or glucose injection. Kind.
The rapamycin nano-sustained release formulation of claim 1, wherein the raw material further comprises a freeze-dried protective agent.
A preparation method of the rapamycin nano sustained-release agent according to any one of claims 1-7, characterized in that it comprises the following steps:

1) Add rapamycin bulk drug and soluble polymer carrier to organic solvent to form organic phase;

2) Inhale the organic phase into the syringe, add 1-10 drops per minute to the aqueous phase, and stir for 30min-3h at room temperature;

3) Recover organic solvents under reduced pressure;

4) Centrifuge for 5-120min, take the supernatant, filter with 0.22-0.45μm membrane to obtain micellar solution;

5) Freeze-dry the micellar solution to obtain a rapamycin nano-sustained release agent.
The preparation method according to claim 8, wherein in step 2), the stirring speed is 500-800 rpm; in step 4), the centrifugal speed is 4000-8000 rpm.
The preparation method according to claim 8, wherein in step 4), 5-10 g of lyophilized protective agent is added to every 100 mL of micellar solution.