CN109091451B

CN109091451B - Oil phase liquid crystal gel precursor preparation of hydrophilic medicine and preparation method thereof

Info

Publication number: CN109091451B
Application number: CN201811050804.7A
Authority: CN
Inventors: 罗亮; 黄丽萍; 孟凡玲
Original assignee: Wuhan Baina Likang Biopharmaceutical Co ltd
Current assignee: Wuhan Baina Likang Biopharmaceutical Co ltd
Priority date: 2018-09-10
Filing date: 2018-09-10
Publication date: 2021-08-13
Anticipated expiration: 2038-09-10
Also published as: CN109091451A

Abstract

The invention belongs to the field of prodrug preparations, and discloses a preparation method of an oil-phase liquid crystal gel prodrug preparation of a hydrophilic drug, which comprises the following steps: preparing a phospholipid dispersion and a hydrophilic drug solution, mixing the phospholipid dispersion and the hydrophilic drug solution to prepare a water phase solution, then preparing an oil phase solution, mixing the oil phase solution and the water phase solution, freeze-drying to prepare a freeze-dried sample, then preparing a mixed oil phase of phospholipid and glyceride, adding the mixed oil phase into the freeze-dried sample, and uniformly mixing to prepare the hydrophilic drug precursor preparation. The obtained hydrophilic prodrug preparation has good stability, can form gel when meeting water, and has good slow release effect. The system has the advantages of simple preparation process, small batch difference, few influencing factors and the like, and is a new technology and a new preparation formulation with high commercial transformation value.

Description

Oil phase liquid crystal gel precursor preparation of hydrophilic medicine and preparation method thereof

Technical Field

The invention belongs to the field of prodrug preparations, and particularly relates to an oil-phase liquid crystal gel prodrug preparation of a hydrophilic drug and a preparation method thereof.

Background

The medicine can be divided into oral preparation, injection preparation, nasal preparation, transdermal preparation and the like through different medicine modes. Different modes of administration require different factors to be considered. For example, for polypeptide protein drugs with large molecular weight, short half-life and poor fat solubility, oral preparations are easy to degrade and inactivate by gastric acid and pepsin; the injection needs to consider the biocompatibility of the carrier, and the like; transdermal preparations are intended to examine the influence of differences in individual and site of action on the absorption of drugs, and the like. In addition, how to improve the storage stability of the drug, reduce the administration frequency, improve the bioavailability of the drug and reduce the side effects of the drug is also extremely important in the development and research of the drug.

Many pharmaceutical active agents are beneficial when maintained at in vivo concentrations within a certain range, and either too high or too low can not achieve the desired effect, and too high a concentration can also lead to toxic side effects. In addition, long-term high frequency administration of some drugs with short half-lives can result in poor patient compliance.

In order to solve the above problems, there is a need to develop a sustained release formulation with good stability, which can effectively control the drug release rate, reduce side effects, reduce the number of times of administration, and improve the compliance of patients.

Lyotropic liquid crystal gels are predominantly ordered systems formed from one or more amphiphilic compounds and a solvent. Due to its internal composition of a crystal lattice, the ordered arrangement of the crystal lattice and the strong interaction between the amphiphilic molecules make it gel-like in appearance. The lyotropic liquid crystal attracts the attention of numerous scientists due to the characteristics of the structure of the lyotropic liquid crystal, is gradually used as a carrier of various medicines, can slowly release various polar medicines in vivo for a long time, ensures that the medicines maintain effective concentration in vivo for a long time, and achieves the purposes of improving the biological activity and reducing the administration times.

In the existing gel preparation technology, phospholipid and glyceride are used as drug carriers, and because of different polarities of different drugs, in order to solve the problem that some drugs are poor in solubility in a system, a method of adding a large amount of solvent and surfactant is generally adopted, after the drugs are dissolved in the phospholipid or the glyceride or the solvent, the components can be mixed to prepare an injection, and water is also added to prepare liquid crystal nanoparticles. The injection can react with water in human body to form gel, and has sustained release effect. The liquid crystal nanoparticles can be used as oral preparations or nasal administration and the like, have good storage stability and have good slow release effect. For example, patent CN108272747A discloses an oil phase liquid crystal gel precursor preparation of a hydrophilic drug and a preparation method thereof, and chinese patent CN108309931A discloses a preparation for treating synovitis and a preparation method thereof. Chinese patent CN108283621A discloses a nasal nano-preparation mometasone furoate liquid crystal gel nanoparticle and a preparation method thereof.

However, for hydrophilic drugs, especially hydrophilic macromolecular drugs such as protein IgG, PD-1, PD-L1 and insulin, the aqueous phase preparation prepared by the method has poor stability and is easy to hydrolyze; the hydrophilic drug oil phase preparation in the current market can not form gel, so that the problem of poor slow release effect generally exists; and the storage stability is poor. Hydrophilic drugs are difficult to dissolve in the oil phase and require the addition of an aqueous phase to dissolve, making it difficult to make a liquid crystal gel precursor formulation with a homogeneous oil phase. Therefore, there is a need to develop a new method for preparing an oil phase preparation of a hydrophilic drug, so that the hydrophilic drug can be prepared into a uniform oil phase preparation, which has a sustained release effect and good stability.

Disclosure of Invention

The invention aims to at least solve one of the problems of the prior art and provides an oil-phase liquid crystal gel precursor preparation of a hydrophilic drug with long-acting slow release effect and a preparation method thereof.

The technical scheme of the invention is as follows: the preparation method of the oil-phase liquid crystal gel precursor preparation of the hydrophilic drug comprises the following steps:

(S1) preparation of phospholipid Dispersion: weighing phospholipid and distilled water, adding the phospholipid and the distilled water into a bottle according to the mass ratio of 1: 3-5, sealing the bottle, and dispersing the bottle on a shaking table to obtain a crude dispersion; ultrasonically dispersing the crude dispersion by a probe under the ice bath condition to form milky phospholipid dispersion;

(S2) preparation of hydrophilic drug solution: weighing hydrophilic drugs, adding water to completely dissolve the hydrophilic drugs, and preparing the hydrophilic drugs into a drug solution of 1-20 mg/ml for later use;

(S3)) preparation of the aqueous phase: uniformly mixing the phospholipid dispersion prepared in the step (S1) and the medicinal solution prepared in the step (S2) in a volume ratio of 1: 3-2: 3 to obtain a water phase;

(S4) preparation of the oil phase: adding 0.01-0.05 wt% of surfactant into glyceride, and uniformly mixing to obtain an oil phase;

(S5) oil phase and water phase are mixed evenly: weighing a proper amount of the oil phase prepared in the step (S4) and adding the oil phase into the water phase prepared in the step (S3) to enable the mass ratio of phospholipid to glyceride in the mixture to be 32: 68-70: 30, uniformly mixing in a vortex mode, and immediately freezing by using liquid nitrogen to obtain a pre-frozen sample;

(S6) preparing a lyophilized sample: freeze-drying the pre-frozen sample to remove the aqueous phase medium to prepare a freeze-dried sample;

(S7) preparing a mixed oil phase: respectively weighing phospholipid and glyceride according to the mass ratio of 32: 68-70: 30, mixing, adding 5-20 wt% of cosolvent, and shaking on a shaking table at 35-40 ℃ until the mixture is completely dissolved to obtain a mixed oil phase;

(S8) preparing a prodrug formulation: and (4) adding a proper amount of the mixed oil phase prepared in the step (S7) into the freeze-dried sample prepared in the step (S6), shaking the mixture on a shaking table at the temperature of 35-40 ℃ until the mixture is completely dissolved to obtain a clear and transparent liquid crystal gel precursor preparation, and sealing the clear and transparent liquid crystal gel precursor preparation for later use.

The scheme of the invention can be used for hydrophilic micromolecular medicaments such as vitamins, hydrophilic macromolecules such as antibody protein, insulin and the like, and can also be used for co-dissolving hydrophilic medicaments of micromolecules and macromolecules. According to the scheme, the phospholipid dispersion and the medicine aqueous solution are mixed and then mixed with the oil phase, the mixture is freeze-dried to remove water, and then the mixed oil phase of phospholipid and glyceride is added, so that the problem that hydrophilic molecules are insoluble in glyceride can be solved, the solubility of hydrophilic medicines is improved, and the prepared prodrug preparation has multilayer protection of oil phospholipid and glyceride and is good in storage stability. The prodrug preparation can be used as an injection, forms gel when entering a human body and meets water, can achieve good slow release effect, prolongs the action time of the drug, reduces the drug administration frequency of the drug and improves the bioavailability of the drug.

When the hydrophilic drug is a glyceride-insoluble macromolecular drug, the hydrophilic drug is preferably one or more of protein IgG, PD-1, PD-L1 or insulin, and the concentration of the hydrophilic drug in the prepared prodrug preparation is 1-20 mg/ml. The invention is particularly suitable for hydrophilic macromolecular drugs in the scheme, can obviously improve the storage stability of the drugs, improve the storage activity and in vivo activity of the drugs, prolong the administration time through slow release and improve the adaptability of patients.

Preferably, the cosolvent is at least one of ethanol, medium-chain fatty acid, long-chain fatty acid, peanut oil, soybean oil, corn oil, castor oil and sesame oil. Further preferably, the cosolvent is ethanol. The preferable cosolvent has good biocompatibility, can improve the solubility of the medicine, has low viscosity, is suitable for injection, and can not cause inflammatory reaction of organisms.

Preferably, the surfactant is a non-ionic surfactant, more preferably tween 80.

Preferably, the glyceride is at least one of glyceryl dioleate, glyceryl trioleate, glyceryl stearate and glyceryl palmitate. Further preferably, the glyceride is glycerol dioleate. The preferred glyceride has lower cost and good biocompatibility, and the complex phospholipid can react with water more quickly to form liquid crystal gel with better stability.

Preferably, the phospholipid is at least one of soybean phospholipid, phosphatidylethanolamine, phosphatidylserine and phosphatidylglycerol. The scheme preferably selects the phospholipid with low cost, good biocompatibility and good amphiphilicity, so that the system can quickly react with water to form liquid crystal gel, and the bioavailability of the medicine is high.

Preferably, the phospholipid consists of 80-90 wt% of soybean phospholipid, 5-10 wt% of phosphatidylethanolamine and 5-10 wt% of phosphatidylserine.

The mixed phospholipid improves the stability of the mixed oil phase, so that the stability of the hydrophilic drug is better.

In conclusion, the beneficial effects of the invention are as follows: the hydrophilic drug is dissolved in the mixed oil phase to form an oil phase preparation, so that the storage stability is good; can be used for injection, and can form gel when injected into the body and meet water, the slow release effect of the gel preparation can maintain the effective concentration of the medicine for a long time, and the administration times are reduced; the raw materials have good biocompatibility and can not cause inflammatory reaction of organisms.

Drawings

FIG. 1 is a graph showing the cumulative release rate of a hydrophilic drug from an oil-phase liquid crystal gel precursor preparation of the hydrophilic drug prepared in examples 1 to 3;

FIG. 2 rheological properties of SPC/GDO samples at different ratios;

FIG. 3 degradation of gels with different ratios of SPC/GDO under the effect of different lipase contents.

Detailed Description

In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is further described in detail with reference to the following embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention. Reagents, methods and apparatus used in the present invention are conventional in the art unless otherwise indicated.

Example 1

The oil-phase liquid crystal gel precursor preparation of the hydrophilic drug of the embodiment is prepared by the following preparation method:

(S1) preparation of phospholipid Dispersion: weighing 1g of phosphatidyl glycerol and 4g of distilled water, adding into a bottle, sealing, and dispersing on a shaking table to obtain a crude dispersion; ultrasonically dispersing the crude dispersion for 20min at 50% (650W) for 1s/3s under the ice bath condition to form milky phospholipid dispersion;

(S2) preparation of hydrophilic drug solution: weighing protein PD-15mg, adding 1.5ml of water, and completely dissolving to prepare a 3.33mg/ml medicinal solution for later use;

(S3)) preparation of the aqueous phase: uniformly mixing 140uL of the phospholipid dispersion prepared in the step (S1) with 300uL of the medicinal solution prepared in the step (S2) to obtain a water phase;

(S4) preparation of the oil phase: adding 0.3mg of Tween 80 into 2.7g of glycerol dioleate, and uniformly mixing to obtain an oil phase;

(S5) oil phase and water phase are mixed evenly: weighing 65mg of the oil phase prepared in the step (S4) and adding the oil phase into the water phase prepared in the step (S3), uniformly mixing by vortex, and immediately freeze-drying by liquid nitrogen to obtain a pre-frozen sample;

(S7) preparing a mixed oil phase: respectively weighing 175mg of phosphatidyl glycerol and 325mg of diglycerol, mixing, adding 25mg of medium-chain fatty acid, and shaking on a shaking table at 37 ℃ until the medium-chain fatty acid is completely dissolved to obtain a mixed oil phase;

(S8) preparing a prodrug formulation: adding 400uL of the mixed oil phase obtained in the step (S7) into the lyophilized sample obtained in the step (S6), shaking on a shaking table at 37 ℃ until the mixed oil phase is completely dissolved to obtain a clear and transparent prodrug preparation, and sealing for later use. The resulting precursor formulation had a protein PD-1 concentration of 2 mg/mL.

After the hydrophilic prodrug preparation prepared by the embodiment is placed at 4 ℃ for 6 months, 9 months and 12 months, no phenomenon occurs in the centrifugation treatment, which indicates that the storage stability of the preparation is good.

The experimental method for the hydrophilic drug release investigation of the hydrophilic prodrug preparation prepared by the invention comprises the following steps: weighing an appropriate amount of oil-phase liquid crystal gel precursor preparation of the hydrophilic drug in a clean and dry EP tube, then adding a certain volume of pH 7.4 phosphate buffer solution, sealing, putting into a constant-temperature shaking table for oscillation release, and setting parameters of the constant-temperature shaking table: the temperature was 37 ℃ and the rotation speed was 100 rpm. Samples were taken at different time points, respectively, and the same volume of release medium was then added to the release system. The released medium is placed at-20 ℃ for standby, and the content of the hydrophilic drug in the released medium is detected by HPLC.

The cumulative release rate of the protein in PD-116 days in the oil phase liquid crystal gel precursor preparation of the hydrophilic medicament is about 10 percent, has good slow release effect, can greatly prolong the medicament administration period and reduce the medicament administration times. The results of the experiment are shown in FIG. 1. The oil phase has different phospholipid and grease proportions and different slow release effects.

Example 2

(S1) preparation of phospholipid Dispersion: weighing 1g of soybean lecithin and 4g of distilled water, adding into a bottle, sealing, and dispersing on a shaking table to obtain a crude dispersion; ultrasonically dispersing the crude dispersion for 20min at 50% (650W) for 1s/3s under the ice bath condition to form milky phospholipid dispersion;

(S2) preparation of hydrophilic drug solution: weighing 5mg of protein IgG, adding 1.5ml of water, and completely dissolving to prepare a medicinal solution of 3.33mg/ml for later use;

(S3)) preparation of the aqueous phase: uniformly mixing 200uL of the phospholipid dispersion prepared in the step (S1) with 300uL of the medicinal solution prepared in the step (S2) to obtain a water phase;

(S5) oil phase and water phase are mixed evenly: weighing 50mg of the oil phase prepared in the step (S4) and adding the oil phase into the water phase prepared in the step (S3), uniformly mixing by vortex, and immediately freeze-drying by liquid nitrogen to obtain a pre-frozen sample;

(S7) preparing a mixed oil phase: respectively weighing 250mg of soybean phospholipid and 250mg of glycerol dioleate, mixing, adding 50mg of ethanol, shaking on a shaking table at 37 ℃ until the soybean phospholipid and the glycerol dioleate are completely dissolved, and mixing oil phases;

(S8) preparing a prodrug formulation: adding 400uL of the mixed oil phase obtained in the step (S7) into the lyophilized sample obtained in the step (S6), shaking on a shaking table at 37 ℃ until the mixed oil phase is completely dissolved to obtain a clear and transparent prodrug preparation, and sealing for later use. The protein IgG concentration of the resulting preparation was 2 mg/mL.

After the hydrophilic prodrug preparation prepared by the embodiment is placed at 4 ℃ for 6 months, 9 months and 12 months, no layering phenomenon exists after centrifugal treatment, and the storage stability of the preparation is good.

In the embodiment, the 16-day cumulative release rate of the protein IgG is about 3.8%, the sustained-release effect is good, the medication period can be greatly prolonged, and the medication frequency is reduced.

Example 3

(S1) preparation of phospholipid Dispersion: weighing 1g of soybean phospholipid, 0.3g of phosphatidylethanolamine and 4g of distilled water, adding into a bottle, sealing, and dispersing on a shaking table to obtain a crude dispersion; ultrasonically dispersing the crude dispersion for 20min at 50% (650W) for 1s/3s under the ice bath condition to form milky phospholipid dispersion;

(S2) preparation of hydrophilic drug solution: weighing 20mg of insulin, adding 1ml of water, and completely dissolving to prepare a medicinal solution of 20mg/ml for later use;

(S3)) preparation of the aqueous phase: uniformly mixing 210uL of the phospholipid dispersion prepared in the step (S1) with 300uL of the medicinal solution prepared in the step (S2) to obtain a water phase;

(S4) preparation of the oil phase: adding 1mg of Tween 80 into 2.7g of the palmitic acid glyceride, and uniformly mixing to obtain an oil phase;

(S5) oil phase and water phase are mixed evenly: weighing 30mg of the oil phase prepared in the step (S4) and adding the oil phase into the water phase prepared in the step (S3), uniformly mixing by vortex, and immediately freeze-drying by liquid nitrogen to obtain a pre-frozen sample;

(S7) preparing a mixed oil phase: respectively weighing 250mg of soybean phospholipid and 250mg of palmitic acid glyceride, mixing, adding 50mg of ethanol, shaking on a shaking table at 37 ℃ until the soybean phospholipid and the palmitic acid glyceride are completely dissolved, and mixing oil phases;

(S8) preparing a prodrug formulation: adding 400uL of the mixed oil phase obtained in the step (S7) into the lyophilized sample obtained in the step (S6), shaking on a shaking table at 37 ℃ until the mixed oil phase is completely dissolved to obtain a clear and transparent prodrug preparation, and sealing for later use. The insulin concentration of the resulting preparation was 10 mg/mL.

In the embodiment, the 16-day cumulative release rate of the insulin is about 11.5%, so that the sustained-release effect is good, the medication period can be greatly prolonged, and the medication frequency is reduced.

Example 4

Compared with the embodiment 2, the phospholipid in the embodiment is composed of 80 wt% of soybean phospholipid, 10 wt% of phosphatidylethanolamine and 10 wt% of phosphatidylserine; the other preparation methods and the used raw materials were the same as in example 2.

Comparative example 1

Compared with example 2, in the preparation method of the oil-phase liquid crystal gel precursor preparation of the hydrophilic drug of the comparative example:

steps (S1) to (S6) are the same as in example 2, and step (S7) is performed by adding 400uL of glycerol dioleate to the lyophilized sample obtained in step (S6), shaking the mixture on a shaker at 37 ℃ until the mixture is completely dissolved to obtain a prodrug preparation, obtaining a clear prodrug preparation, and sealing the prodrug preparation for use. The protein IgG concentration of the resulting preparation was 2 mg/mL.

The hydrophilic prodrug preparation prepared by the comparative example has no delamination after 6 months and 9 months of storage at 4 ℃, the delamination occurs after 12 months, and the delamination also occurs after 9 months of centrifugal treatment, which indicates that the storage stability of the preparation is poorer than that of the preparation prepared by the example.

Comparative example 2

The comparative example provides a method of preparing an oil phase liquid crystal gel precursor formulation of a hydrophilic drug as follows:

(S3)) preparation of the aqueous phase: uniformly mixing 200uL of the phospholipid dispersion prepared in the step (S1) with 300uL of the medicinal solution prepared in the step (S2) to obtain a water phase; immediately freeze-drying the liquid nitrogen to obtain a pre-frozen sample;

(S4) preparing a lyophilized sample: freeze-drying the pre-frozen sample to remove the aqueous phase medium to prepare a freeze-dried sample;

(S5) preparing a mixed oil phase: respectively weighing 250mg of soybean phospholipid and 250mg of glycerol dioleate, mixing, adding 50mg of ethanol, completely dissolving and uniformly mixing in a 45 ℃ oven, and mixing oil phases;

(S6) preparing a prodrug formulation: adding 400uL of the mixed oil phase obtained in the step (S5) into the lyophilized sample obtained in the step (S4), shaking on a shaking table at 37 ℃ until the mixed oil phase is completely dissolved to obtain a prodrug preparation, and sealing for later use. The protein IgG concentration of the resulting preparation was 2 mg/mL.

The hydrophilic prodrug preparation prepared by the comparative example has no delamination after being placed at 4 ℃ for 6 months, 9 months and 12 months, and the delamination phenomenon occurs after 12 months of centrifugal treatment, which indicates that the storage stability of the preparation is poorer than that of the preparation in the examples.

Comparative example 3

(S5) preparation of the oil phase: adding 0.3mg of Tween 80 into 2.7g of glycerol dioleate, and uniformly mixing to obtain an oil phase;

(S6) preparing a prodrug formulation: and (4) adding 400uL of the oil phase prepared in the step (S5) into the freeze-dried sample prepared in the step (S4), uniformly mixing to obtain a prodrug preparation, and sealing for later use. The protein IgG concentration of the resulting preparation was 2 mg/mL.

Comparative example 4

A method for preparing a hydrophilic drug, comprising the steps of: mixing and dissolving 20mg of insulin, 450mg of phospholipid, 450mg of glycerol dioleate, 90mg of ethanol and 30ug of Tween 80 to obtain the insulin liquid crystal gel precursor preparation.

The insulin liquid crystal gel precursor preparation prepared by the comparative example has a layering phenomenon after being placed at 4 ℃ for 6 months, which shows that the storage stability of the preparation of the comparative example is poorer than that of the preparations of the examples of each group.

Comparative example 5

The prodrug preparation of this comparative example was a solution prepared by directly dissolving protein IgG in water to a concentration of 2 mg/mL.

Comparative example 6

The prodrug preparation of this comparative example was prepared by dissolving insulin directly in water to give a solution with a concentration of 10 mg/mL.

IgG protein released in the IgG liquid crystal gel precursor preparation release degree experiments in the comparative example, the example 2 and the example 4 is precipitated by ethanol, an equal amount of IgG is taken as a sample from the comparative example, the example 2 and the example 4, an enzyme-linked immunosorbent assay is carried out by adopting an immunoglobulin G (IgG) ELISA kit of Shanghai Tong-Wei-Tu-biological science and technology Co., Ltd, the specific steps are the same as the kit operation instruction, and the experimental results are shown in Table 1.

TABLE 1 results of ELISA adsorption test for IgG of different groups

As can be seen from Table 1, for the IgG liquid crystal gel precursor preparation, the preparations of examples 2 and 4 of the invention release IgG protein with bioactivity in higher content than the comparative example, which indicates that the IgG protein released by the preparation of the invention has complete structure and stable activity, and probably because the phospholipid and glyceride form multilayer protection effect on the IgG protein in the method of the invention, the stability of the IgG protein is good, and the loss caused by degradation or denaturation of the IgG protein is reduced in the processes of freeze-drying and redissolution.

The insulin released from the insulin liquid crystal gel precursor preparation in example 3 and comparative example in the release test was precipitated with ethanol, and the same mass of insulin as in comparative example and example 3 was sampled and measured by 1211 insulin bioassay in accordance with pharmacopoeia of the people's republic of china (2015, fourth part), the experimental results of which are shown in table 2.

TABLE 2 bioassay of insulin for different groups

Group of	Example 3	Comparative example 4	Comparative example 6
				Blood sugar value (mmol/L)	6.3±1.0	7.1±1.2	7.6±1.1

As can be seen from Table 2, the blood glucose lowering effect of the formulation of example 3 of the present invention is better than that of the formulations of comparative examples 4 and 6, indicating that the biological activity of insulin of the formulation of the present invention is higher, probably because the phospholipid and glyceride have protective effect on insulin in the method of the present invention, so that the stability of insulin is good, and the activity loss caused by degradation or denaturation of insulin is reduced in the processes of freeze-drying and reconstitution.

The rheological properties of the samples were also tested after mixing soya lecithin (SPC) and Glycerol Dioleate (GDO) in a ratio of 32:68, 50:50 and 70:30, respectively, and the elastic modulus (G') and viscous modulus (G ") of the gels were examined using a rheometer, the results of which are shown in fig. 2, and it can be seen that as the SPC/GDO ratio increases, the elastic modulus of the formed gels increases first and then decreases.

In addition, the degradation of samples of Soybean Phospholipid (SPC) and diglycerol oleate (GDO) at different ratios under the action of different lipase contents after gel formation is also examined. Soybean Phospholipid (SPC) and Glycerol Dioleate (GDO) were tested for weight change at ratios of 35:65, 50:50, and 70:30, respectively, and with addition of 0%, 2%, and 5% lipase, respectively. The results are shown in FIG. 3. It can be seen that under the action of lipase, the gel firstly absorbs water and swells, the mass is increased, and then the gel is degraded, and the mass is reduced; and the degradation speed is accelerated along with the increase of the content of the lipase.

The results show that the carrier in the oil-phase liquid crystal gel precursor preparation of the hydrophilic drug can be degraded in vivo, the preparation can form gel after entering human body and meeting water, the in-vitro 16-day cumulative release rate of the drug in the gel is not more than 15%, the hydrophilic drug has long-term sustained release, and the stability of the hydrophilic drug can be obviously improved compared with the oil-phase preparation and the water-phase preparation prepared by the method in the comparative example.

Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that changes, modifications, substitutions and alterations can be made in these embodiments without departing from the principles and spirit of the invention, the scope of which is defined in the appended claims and their equivalents.

Claims

1. The preparation method of the oil-phase liquid crystal gel precursor preparation of the hydrophilic drug is characterized by comprising the following steps:

(S8) preparing a prodrug formulation: adding a proper amount of the mixed oil phase prepared in the step (S7) into the freeze-dried sample prepared in the step (S6), shaking the mixture on a shaking table at the temperature of 35-40 ℃ until the mixture is completely dissolved to obtain a clear and transparent liquid crystal gel precursor preparation, and sealing the clear and transparent liquid crystal gel precursor preparation for later use;

the hydrophilic drug is one or more of macromolecular proteins IgG, PD-1, PD-L1 or insulin which are insoluble in glyceride, and the concentration of the hydrophilic drug in the prepared prodrug preparation is 1-20 mg/ml;

the cosolvent is at least one of ethanol or medium-chain fatty acid;

the glyceride is at least one of diglyceride, triolein, stearin and palmitin;

the phospholipid is at least one of soybean phospholipid, phosphatidylethanolamine, phosphatidylserine and phosphatidylglycerol.

2. The method for preparing an oil-phase liquid crystal gel precursor preparation of a hydrophilic drug according to claim 1, wherein the cosolvent is ethanol.

3. The method for preparing an oil-phase liquid crystal gel precursor preparation of a hydrophilic drug according to claim 1, wherein the glyceride is glycerol dioleate.

4. The method for preparing an oil-phase liquid crystal gel precursor preparation of a hydrophilic drug according to claim 1, wherein the phospholipid comprises 80-90 wt% of soybean phospholipid, 5-10 wt% of phosphatidylethanolamine, and 5-10 wt% of phosphatidylserine.

5. A prodrug formulation prepared by the process according to any one of claims 1 to 4.