WO2019007019A1 - Psoralen polymernanoparticle preparation and preparation method therefor - Google Patents

Abstract

A psoralen polymernanoparticle preparation and a preparation method therefor, relating to the technical field of polymernanoparticle preparations. The psoralen polymernanoparticle preparation with a high encapsulation efficiency is prepared by using a proper amount of psoralen, polylactide-glycolide, soybean phospholipid, and polyethylene glycol 1000 vitamin E succinate as raw materials, and in particular by using polyethylene glycol 1000 vitamin E succinate as a surfactant and soybean phospholipid as a lipid. By optimizing the preparation conditions of the psoralen polymernanoparticle preparation, the encapsulation efficiency of the psoralen polymernanoparticle preparation can be further improved. The encapsulation efficiency is up to 84.97%.

Description

Psoralen polymer nanoparticle preparation and preparation method thereof

The present application is based on a Chinese patent application filed on Jan. 6, 2017, the entire disclosure of which is hereby incorporated by reference.

Technical field

The present application relates to the technical field of polymer nanoparticle preparations, and in particular to a psoralen polymer nanoparticle preparation and a preparation method thereof.

Background technique

Tumor is one of the major diseases that endanger human health today. Tumor invasion and metastasis is the most common biological behavior and the most essential feature of malignant tumors, and also a key factor affecting the survival and recovery of cancer patients. Therefore, anti-tumor drugs are the subject of constant research and development by those skilled in the art. In recent years, people have turned to other ways to find an effective method to reverse the MDR of tumor cells. Due to the small volume and special structure of nanoparticles, they show unique advantages in improving drug absorption, distribution, metabolism, excretion and toxicity. Therefore, the preparation of nano drug delivery systems and nanoparticles has received increasing attention. The nano drug delivery system includes nano-liposomes, solid lipid nanoparticles (SLN), polymer nanoparticles (PLN), nanospheres, nanocapsules and microemulsions, and the particle size of the nano drug delivery system is generally Between 10-500 nm, the nano-drug delivery system can be used to reverse the tumor MDR effect. Nanoliposomes have a biofilm-like structure, and lipophilic or hydrophilic drugs can be designed as liposomes, which have passive targeting properties. The use of polymers to modify nanoliposomes can achieve different therapeutic needs, such as Targeting, sustained release, environmental dependence, etc. Microemulsion is an optically isotropic and thermodynamically stable liquid-liquid dispersion system composed of an emulsifier, a co-emulsifier, an oil phase and an aqueous phase. The drug has good dispersibility in microemulsion, and it has an increase in poorly soluble drugs. Solubilization, which increases bioavailability. SLN is a nano-preparation prepared from lipid materials, and SLN can promote the penetration of P-gp-enriched blood-brain barrier. Many studies have focused on the preparation of chemotherapeutic drug SLN and its mechanism of action. Anti-P-gp small molecule inhibitors (such as verapamil, cyclosporin A, GG918, etc.) and anti-tumor drugs are used together to prepare SLN. PLN is a hydrophilic micelle prepared by polymerization of sodium bis(2-ethylhexyl)sulfosuccinate as a monomer. Due to the potential biotoxicity of non-degradable materials, polymethyl cyanide is gradually being used. A acrylate, a polystyrene, a polyamide, or the like is used as a material to prepare a biodegradable PLN or the like which can be directed to a lysosome.

Research and products on chemotherapeutic drug nanoparticles have been reported and marketed. For example, Abraxane is the first non-dissolved nano-albumin-binding chemotherapeutic drug, which is a paclitaxel-coated nanoparticle coated with albumin for the treatment of breast cancer metastasis; Gliadelwafer is a poly Phenylpropanoid 20 encapsulates the nanoparticles of carmustine for the treatment of highly differentiated malignant glioma; DaunoXome is a daunorubicin liposome for the treatment of Kaposi's sarcoma; Myocet is a doxorubicin liposome , in combination with cyclophosphamide for the treatment of metastatic breast cancer; DOXIL is doxorubicin liposome for the treatment of metastatic ovarian cancer; doxorubicin galactosamine and N-(2-hydroxypropyl) The copolymer of methacrylamide, PK2, is used to treat liver cancer. The various pharmaceutical preparations of the above drug-loading system have certain effects of reversing MDR activity, but there are also some problems, such as low drug loading, poor stability and easy leakage.

The application of biodegradable PLN can effectively improve the stability of the drug, thereby functioning as a sustained release and a controlled release. Currently available biodegradable carrier materials mainly include biodegradable polymers and natural macromolecular systems. Biodegradable polymers include polylactide (PLA), polyglycolide (PLG), and polylactide. - glycolide (PLGA), polycaprolactone (PCL), polyorthoester (POE), polycyanoacrylate (PACA), polyvinylpyrrolidone (PVP), etc.; natural macromolecular systems include proteins, Polysaccharides, gelatin, polyacrylamide starch, chitin and derivatives, sodium alginate, gelatin, albumin, lecithin, cholesterol, and the like. As for the above polymer, polylactide-glycolide (PLGA) is a polymer compound obtained by polymerizing lactic acid and glycolic acid, and has the advantages of low toxicity, good granulation property and biocompatibility, and PLGA carrier material can be used. The water-soluble system is degraded by the cleavage of the ester bond, and the lactic acid and glycolic acid produced by the degradation are further degraded into water and carbon dioxide in the body, and the release rate of the encapsulated drug is regulated by the PLGA self-degradation rate, so that the drug can be used for a long time. It is released at a steady rate and maintains a stable blood concentration. PLGA can increase the water solubility of fat-soluble drugs and improve the bioavailability of fat-soluble drugs. PLGA has been approved by the US FDA for use in pharmaceutical carriers and other fields. By changing the ratio of lactic acid and glycolic acid in PLGA, the degradation rate of PLGA can be affected and changed. . At present, the preparation method of PLGA nanoparticle is more mature in the rehydration solvent evaporation method. The research on the preparation method and physicochemical properties of the chemotherapeutic drug is only reported in the PLGA nanoparticle preparation research, and the PLGA nanocarrier entrapped drug resistance reversal agent. And the application of anti-tumor MDR has not been reported.

There are significant differences in the encapsulation efficiency and stability of PLN preparations for different active drugs. How to improve the encapsulation efficiency and stability of active drugs for specific PLN preparations is still a hot spot, focus and difficulty in current research. .

Application content

The present application is directed to the problem of low encapsulation efficiency and poor stability of the existing PLN preparation with psoralen as an active drug, and provides a psoralen polymerization with high encapsulation efficiency by optimizing components and preparation conditions. Nanoparticle preparation, and preparation method of the psoralen polymer nanoparticle preparation.

To achieve the above object, the present application adopts the following technical solutions.

A psoralen polymer nanoparticle preparation is prepared from the following parts by weight: 1-4 parts of psoralen, 10-60 parts of polylactide-glycolide, 30-110 parts of soybean phospholipid, 5 -11 parts of polyethylene glycol 1000 vitamin E succinate.

Preferably, the psoralen polymer nanoparticle preparation is prepared from the following parts by weight: 2-3 parts of psoralen, 40-50 parts of polylactide-glycolide, 70-110 Parts of soybean phospholipid, 5-7 parts of polyethylene glycol 1000 vitamin E succinate.

More preferably, the psoralen polymer nanoparticle preparation is made up of the following parts by weight: 2 parts of psoralen, 50 parts of polylactide-glycolide, 70 parts of soybean phospholipid, 6 Part of polyethylene glycol 1000 vitamin E succinate.

More preferably, the psoralen polymer nanoparticle preparation is characterized by being made up of the following parts by weight: 2 parts of psoralen, 50 parts of polylactide-glycolide, 110 A portion of soy lecithin, 6 parts of polyethylene glycol 1000 vitamin E succinate.

Preferably, the mass ratio of lactic acid to glycolic acid in the above polylactide-glycolide is 1:1.

The preparation method of the psoralen polymer nanoparticle preparation described above comprises the following steps:

S1, psoralen, polylactide-glycolide, soybean phospholipid and polyethylene glycol 1000 vitamin E succinate are weighed according to the weight parts, and used.

Dissolving polylactide-glycolide in an organic solvent, and then adding psoralen to the organic solvent to form an organic phase; standby; the volume ratio of the mass of the polylactide-glycolide to the organic solvent is 5 -50g: 1.6-2.4 mL.

Preferably, the organic solvent is ethanol or acetone.

S2, soy phospholipid and polyethylene glycol 1000 vitamin E succinate are dissolved in an aqueous ethanol solution to form an aqueous phase, which is reserved; the volume ratio of the soybean phospholipid to the aqueous ethanol solution is 1-5 g: 1 mL.

Preferably, the mass fraction of ethanol in the aqueous ethanol solution is 4%.

S3, injecting the organic phase into the aqueous phase to form a mixed solution; heating the mixed solution to 60-80 ° C, and stirring the mixed solution at a flow rate of 10-30 rpm for 30-120 min to form an emulsion.

Preferably, the mixture is heated to 75 ° C and stirred.

Preferably, the mixture is stirred at 20 rpm for 1 h.

Preferably, the organic phase is uniformly injected into the aqueous phase with a syringe.

S4. Pour the emulsion into ice water, and stir the mixture at 10-30 rpm for 30-120 min to form a nanoparticle solution.

Preferably, the mixture is stirred at 20 rpm for 1 h.

After the S5 and the nanoparticle solution were centrifuged, the supernatant liquid was filtered through a 0.45 μm aqueous filter to obtain a psoralen polymer nanoparticle preparation.

Preferably, the nanoparticle solution is centrifuged at 1000 rpm for 5 min.

The psoralen polymer nanoparticle preparation was stored in an environment of 4 °C.

Compared with the prior art, the beneficial effects of the present application are: the present application uses an appropriate amount of psoralen, polylactide-glycolide, soybean phospholipid and polyethylene glycol 1000 vitamin E succinate as raw materials, especially The psoralen polymer nanoparticle preparation with high encapsulation efficiency can be prepared by using polyethylene glycol 1000 vitamin E succinate as a surfactant and soybean phospholipid as a lipid. By optimizing the preparation conditions of the psoralen polymer nanoparticle preparation, the encapsulation efficiency of the psoralen polymer nanoparticle preparation can be further improved, and the encapsulation efficiency can be as high as 84.97%.

Detailed ways

In order to more fully understand the technical content of the present application, the technical solutions of the present application are further introduced and illustrated below in conjunction with specific embodiments.

Example 1-77

The preparation method of the psoralen polymer nanoparticle preparation provided in Example 1-77 is as follows (the amounts and process parameters of each material in each embodiment are shown in Tables 1 and 2):

(1) Reclaiming: psoralen (PSO), polylactide-glycolide (PLGA), soybean phospholipid and polyethylene glycol 1000 vitamin E succinate (TPGS) were weighed according to the amount of each component. ,spare.

(2) Preparation of organic phase: PLGA is dissolved in an organic solvent, and then PSO is added to an organic solvent to form an organic phase;

(3) Water-making phase: Soybean phospholipid and TPGS are dissolved in an aqueous ethanol solution to form an aqueous phase, which is reserved.

(4) Emulsion: The syringe uniformly injects the organic phase into the aqueous phase to form a mixed solution; after heating the mixture to 60-80 ° C, the mixture is stirred at 10-30 rpm for 30-120 min to form an emulsion.

(5) Preparation of nanoparticle solution: The emulsion was poured into ice water, and the mixture was stirred at 10 to 30 rpm for 30 to 120 minutes to form a nanoparticle solution.

(6) Nanoparticle preparation: The obtained nanoparticle solution was centrifuged at 1000 rpm for 5 min, and the supernatant liquid was filtered through a 0.45 μm aqueous filter to obtain a psoralen polymer nanoparticle preparation.

(7) Preservation: The obtained psoralen polymer nanoparticle preparation was stored in an environment of 4 °C.

Table 1 The amount of each material in Example 1-77

In Table 1, PLGA refers to polylactide-glycolide having a mass ratio of lactic acid to glycolic acid of 1:1.

Table 2 Preparation parameters in Examples 1-77

Example 78

The preparation method of the psoralen polymer nanoparticle preparation provided by the embodiment is basically the same as the preparation method of the embodiment 4, and the difference is:

Step (1) was taken as: 3.3 mg of psoralen, 52.5 mg of polylactide-glycolide (same as in Example 4), 32.5 mg of soybean phospholipid, 8.3 mg of synthetic phospholipid (DLPC) and 100 mg, respectively. Tween-80, spare.

Step (3) The aqueous phase is prepared by dissolving soybean phospholipid, synthetic phospholipid and Tween-80 in 20 mL of a 4 wt.% aqueous solution of ethanol to form an aqueous phase, which is ready for use.

Example 79

The preparation method of the psoralen polymer nanoparticle preparation provided by the embodiment is basically the same as the preparation method of the embodiment 4, and the difference is:

Step (1) was taken as: 3.3 mg of psoralen, 52.3 mg of polylactide-glycolide (same as in Example 4), 6.4 mg of synthetic phospholipid (DLPC) and 100 mg of dimethylaminocarb, respectively. Borane (DMAB), spare.

Step (3) The aqueous phase is prepared by dissolving the synthetic phospholipid and dimethylaminoborane in 20 mL of a 4 wt.% aqueous solution of ethanol to form an aqueous phase, which is ready for use.

Example 80

The preparation method of the psoralen polymer nanoparticle preparation provided by the embodiment is basically the same as the preparation method of the embodiment 4, and the difference is:

Step (1) was taken as: 3.2 mg of psoralen, 47.3 mg of polylactide-glycolide (same as in Example 4), 6.1 mg of synthetic phospholipid (DLPC), 32.5 mg of soybean phospholipid and 108.8, respectively. Mg dimethylamino borane (DMAB), ready for use.

Step (3) The aqueous phase is prepared by dissolving soybean phospholipid, synthetic phospholipid and dimethylaminoborane in 20 mL of a 4 wt.% aqueous solution of ethanol to form an aqueous phase, which is ready for use.

Example 81

The preparation method of the psoralen polymer nanoparticle preparation provided by the embodiment is basically the same as the preparation method of the embodiment 4, and the difference is:

Step (1) was taken as: 3.2 mg of psoralen, 51.2 mg of polylactide-glycolide (same as in Example 4), 6.3 mg of synthetic phospholipid (DLPC), 32.6 mg of soybean phospholipid and 21.8, respectively. Mg (MTC), spare.

Step (3) The aqueous phase is prepared by dissolving soybean phospholipid, synthetic phospholipid and MTC in 20 mL of a 4 wt.% aqueous solution of ethanol to form an aqueous phase, which is ready for use.

Example 82

The preparation method of the psoralen polymer nanoparticle preparation provided by the embodiment is basically the same as the preparation method of the embodiment 4, and the difference is:

Step (1) was taken as: 3.1 mg of psoralen, 49.2 mg of polylactide-glycolide (same as in Example 4), 6.1 mg of synthetic phospholipid (DLPC), and 100 mg of dimethylaminocarb, respectively. Borane (DMAB) and 20 mg (MTC), ready for use.

Step (3) The aqueous phase is prepared by dissolving the synthetic phospholipid, dimethylaminoborane and MTC in 20 mL of a 4 wt.% aqueous solution of ethanol to form an aqueous phase, which is ready for use.

Example 83

The preparation method of the psoralen polymer nanoparticle preparation provided by the embodiment is basically the same as the preparation method of the embodiment 4, and the difference is:

Step (1) was taken as: 3.2 mg of psoralen, 47.3 mg of polylactide-glycolide (same as in Example 4), 30 mg of soybean phospholipid and 100 mg of dimethylaminoborane (DMAB), respectively. ),spare.

Step (3) The aqueous phase is prepared by dissolving soybean phospholipid and dimethylaminoborane in 20 mL of a 4 wt.% aqueous solution of ethanol to form an aqueous phase, which is ready for use.

Example 84

The preparation method of the psoralen polymer nanoparticle preparation provided by the embodiment is basically the same as the preparation method of the embodiment 4, and the difference is:

Step (1) was taken as: 3 mg of psoralen, 50.1 mg of polylactide-glycolide (same as in Example 4), 6 mg of synthetic phospholipid (DLPC) and 11 mg of polyethylene glycol 1000 vitamin E, respectively. Succinate (TPGS1000), ready for use.

Step (3) The aqueous phase is prepared by dissolving the synthetic phospholipid and polyethylene glycol 1000 vitamin E succinate in 20 mL of a 4 wt.% aqueous solution of ethanol to form an aqueous phase, which is ready for use.

The encapsulation efficiency, particle diameter, potential, and PDI (Polymer Dispersibility Index) of the psoralen polymer nanoparticle preparation prepared in the above Examples 1-84 are shown in Table 3 below.

Table 3 Encapsulation efficiency, particle size, potential and PDI of the psoralen polymer nanoparticle preparation prepared in Examples 1-84

Example	Particle size nm	PDI	Potential	Encapsulation rate%
1	183.3	0.159	-26.2	59.6
2	190.4	0.144	-30.3	83.1
3	175.1	0.148	-32.6	83.8
4	173.7	0.139	-35.7	84.0
5	172	0.131	-34.3	69.8
6	223.7	0.151	-36.2	85.0
7	183.4	0.151	-35.5	59.8
8	194.3	0.146	-36.1	56.0

9	198.4	0.154	-34.6	51.6
10	181.1	0.162	-34.9	37.6
11	183.4	0.168	-37.1	56.0
12	173.2	0.126	-29.8	69.7
13	194.1	0.131	-27.9	71.3
14	214.6	0.213	-34.0	49.7
15	173.4	0.138	-29.4	76.2
16	176.9	0.129	-29.0	79.2
17	175.5	0.127	-31.5	80.3
18	174.9	0.136	-34.1	77.9
19	206.4	0.201	-36.2	52.4
20	170.0	0.137	-31.2	78.6
twenty one	169.8	0.131	-28.4	80.1
twenty two	179.9	0.129	-29.3	81.9
twenty three	174.9	0.126	-27.9	79.0
twenty four	231.0	0.218	-30.6	47.1
25	169.6	0.132	-28.6	78.6
26	171.3	0.135	-29.9	79.1
27	182.1	0.129	-31.1	76.4
28	169.9	0.136	-30.6	78.6
29	180.0	0.130	-34.1	77.7
30	167.3	0.126	-30.6	80.1
31	249.6	0.191	-39.1	30.1
32	261.3	0.187	-34.6	29.4
33	176.2	0.231	-30.1	60.1
34	180.3	0.206	-32.4	58.7
35	186.4	0.214	-34.9	56.1
36	209.7	0.231	-37.3	28.4
37	194.3	0.201	-30.5	59.7
38	176.9	0.161	-32.6	63.8
39	179.0	0.153	-34.6	64.1
40	183.2	0.193	-30.4	60.9
41	241.0	0.200	-36.4	29.9
42	189.3	0.263	-30.9	36.4
43	183.6	0.149	-29.1	57.9
44	176.4	0.143	-27.3	54.1
45	187.9	0.134	-30.6	59.2
46	185.1	0.129	-27.4	60.7
47	200.3	0.203	-36.8	40.0

48	215.6	0.194	-31.6	41.5
49	176.3	0.143	-30.6	63.1
50	191.0	0.153	-31.4	64.9
51	183.4	0.126	-35.0	59.7
52	172.4	0.171	-37.0	64.9
53	165.3	0.134	-31.9	68.1
54	171.1	0.146	-29.7	64.3
55	168.3	0.135	-29.1	66.6
56	161.3	0.149	-30.6	68.8
57	173.5	0.141	-31.9	61.9
58	169.3	0.144	-30.0	67.3
59	174.6	0.139	-35.1	59.4
60	203.1	0.181	-39.1	42.6
61	176.9	0.176	-30.9	58.3
62	183.1	0.182	-31.1	54.9
63	186.1	0.149	-28.6	60.3
64	179.3	0.164	-29.9	58.3
65	191.0	0.153	-30.9	38.7
66	184.6	0.143	-31.5	34.6
67	190.3	0.14	-30.8	64.5
68	179.9	0.138	-34.6	64.9
69	184.1	0.143	-36.4	63.8
70	180.3	0.141	-34.0	69.1
71	183.0	0.129	-32.5	64.3
72	176.8	0.134	-33.3	65.7
73	189.3	0.136	-34.9	61.3
74	186.4	0.124	-37.0	66.6
75	176.4	0.126	-34.6	59.9
76	174.6	0.130	-30.8	60.3
77	186.3	0.136	-34.6	64.3
78	164.2	0.131	-37.6	57.7
79	204.7	0.194	-31.8	31.58
80	176.1	0.220	-36.9	53.3
81	184.0	0.171	-34.5	56.5
82	172.8	0.217	-42.6	77.3
83	166.8	0.134	-40.4	57.4
84	197.2	0.137	-36.6	52.3

The encapsulation efficiency, particle size, potential and PDI of the psoralen polymer nanoparticle preparation prepared in the above Examples 1-84 can be seen, the encapsulation efficiency, particle diameter, potential and the psoralen polymer nanoparticle preparation. The properties of PDI and the like are affected by many factors. Not only the main components of the preparation of the psoralen polymer nanoparticle preparation will significantly affect its properties, the type of solvent used in the preparation of the organic phase and the preparation of the aqueous phase, and the temperature during the preparation of the emulsion or even the stirring. The speed and time will have a significant impact on its properties, so it is necessary to balance the various factors in order to prepare a psoralen polymer nanoparticle preparation with high encapsulation efficiency and high stability.

The above description of the technical content of the present application is further described by way of example only, so that the reader can understand the present invention more easily, but the embodiment of the present application is not limited thereto, and any technology extension or re-creation according to the application is subject to Protection of this application.

Claims (10)

1. A psoralen polymer nanoparticle preparation, which is prepared from the following parts by weight:

   1-4 parts of psoralen, 10-60 parts of polylactide-glycolide, 30-110 parts of soybean phospholipid, 5-11 parts of polyethylene glycol 1000 vitamin E succinate.
2. A psoralen polymer nanoparticle preparation according to claim 1, which is prepared from the following parts by weight: 2-3 parts of psoralen, 40-50 parts of polylactide-B Lactide, 70-110 parts of soy lecithin, 5-7 parts of polyethylene glycol 1000 vitamin E succinate.
3. A psoralen polymer nanoparticle preparation according to claim 2, which is prepared from the following parts by weight: 2 parts of psoralen, 50 parts of polylactide-glycolide, 70 A portion of soy lecithin, 6 parts of polyethylene glycol 1000 vitamin E succinate.
4. A psoralen polymer nanoparticle preparation according to claim 2, which is prepared from the following parts by weight: 2 parts of psoralen, 50 parts of polylactide-glycolide, 110 A portion of soy lecithin, 6 parts of polyethylene glycol 1000 vitamin E succinate.
5. A method for preparing a psoralen polymer nanoparticle preparation according to claim 1, comprising the steps of:

   S1, weighing psoralen, polylactide-glycolide, soybean phospholipid and polyethylene glycol 1000 vitamin E succinate according to the weight parts, respectively;

   Dissolving polylactide-glycolide in an organic solvent, and then adding psoralen to the organic solvent to form an organic phase, which is used; the volume ratio of the mass of the polylactide-glycolide to the organic solvent is 5 -50g: 1.6-2.4mL;

   S2, the soybean phospholipid and the polyethylene glycol 1000 vitamin E succinate are dissolved in an aqueous solution of ethanol to form an aqueous phase, which is reserved; the volume ratio of the mass of the soybean phospholipid to the aqueous ethanol solution is 1-5 g: 1 mL;

   S3, the organic phase is injected into the water phase to form a mixed solution; the mixture is heated to 60-80 ° C, and the mixture is stirred at a frequency of 10-30 rpm for 30-120 min to form an emulsion;

   S4, the emulsion is poured into ice water, and the mixture is stirred at 10-30 rpm for 30-120 min to form a nanoparticle solution;

   After the S5 and the nanoparticle solution were centrifuged, the supernatant liquid was filtered through a 0.45 μm aqueous filter to obtain a psoralen polymer nanoparticle preparation.
6. The method for preparing a psoralen polymer nanoparticle preparation according to claim 5, wherein in the step S1, the organic solvent is ethanol or acetone.
7. The method for preparing a psoralen polymer nanoparticle preparation according to claim 5, wherein in step S2, the mass fraction of ethanol in the aqueous ethanol solution is 4%.
8. The method for producing a psoralen polymer nanoparticle preparation according to claim 5, wherein in the step S3, the mixture is heated to 75 ° C and stirred.
9. The method for producing a psoralen polymer nanoparticle preparation according to claim 8, wherein in step S3, the mixed solution is stirred at a rotational speed of 20 rpm for 1 hour.
10. The method for producing a psoralen polymer nanoparticle preparation according to claim 5, wherein in step S4, the mixture is stirred at a rotational speed of 20 rpm for 1 hour.