Preparation of docetaxel-stearate triglyceride prodrug and its lipid formulation

Abstract

The present invention belongs to the field of medical technology, and relates to docetaxel-stearic acid triglyceride prodrug and its lipid preparation for lymphatic-mediated transport and its application in the preparation of oral chemotherapy drugs. The present invention provides a lipid preparation of docetaxel-stearic acid triglyceride prodrug, comprising docetaxel-stearic acid triglyceride prodrug and adjuvants, wherein the adjuvants include phospholipid emulsifiers, short-chain alcohols or ethers emulsifiers, and liquid oil phases, wherein by weight percentage, the liquid oil phase accounts for 30%-85% of the lipid preparation, the phospholipid emulsifier accounts for 9%-45% of the lipid preparation, and the short-chain alcohols or ethers emulsifiers account for 5%-25% of the lipid preparation; docetaxel-stearic acid triglyceride prodrug accounts for 1-10% of the total weight of the adjuvant. The preparation method is simple, easy to industrialize, and highly repeatable; the obtained lipid preparation is stable, and can improve the docetaxel efficacy and reduce toxicity;.

Description

Technical Field

[0001] The present invention belongs to the field of medical technology, and relates to docetaxel-stearate triglyceride prodrugs and lipid preparations thereof, and specifically to lymphatic-mediated transport docetaxel-stearate triglyceride prodrugs and lipid preparations thereof, and applications thereof in the preparation of oral chemotherapy drugs. Background Art

[0002] Nowadays, the global cancer burden is increasing year by year. Docetaxel (DTX) is widely used in the clinical treatment of various tumors as a first-line broad-spectrum anticancer chemotherapy drug. Docetaxel is clinically administered by intravenous injection, but the commercially available intravenous solution uses Tween-80 and ethanol for solubilization, which brings about excipient-related toxic side effects and limits clinical application. Oral chemotherapy has the advantages of high patient compliance, convenient administration, and low treatment cost. However, docetaxel has low solubility, P-glycoprotein efflux, and severe first-pass effect, which makes its oral bioavailability extremely low. There is currently no oral docetaxel formulation on the market. Therefore, the development of oral docetaxel formulations with low first-pass effect and high oral bioavailability remains a research hotspot.

[0003] Lymphatic transport of oral drugs can avoid the first-pass effect and is an effective way to improve oral absorption. One strategy to improve lymphatic transport is to design a triglyceride-like prodrug of the drug. The fatty acids at positions 1 and 3 of long-chain triglycerides are specifically hydrolyzed by pancreatic lipase, and the fatty acid at position 2 is almost not hydrolyzed. The fatty acid at position 2 is replaced by a drug, so that the prodrug simulates the digestion process of triglycerides in the intestine, enters the intestinal epithelial cells in the form of 2-monoglyceride prodrugs for re-esterification, participates in the assembly of lipoproteins, and then promotes lymphatic transport. Many researchers have made examples of triglyceride-like prodrugs, but the structures taught by these examples are almost ineffective in improving oral administration of drugs. An important reason is the lack of a cleavable linking chain, which results in the non-release of the parent drug. In CN106715456A, the inventor introduced a self-eliminating linking chain to promote the systemic release of the parent drug testosterone. However, for docetaxel, a chemotherapy drug, a large amount of systemic exposure will bring serious side effects. Therefore, the linking chain is of no guiding significance for the design of docetaxel oral delivery system.

[0004] In the prior art, medical workers prepare docetaxel into triglyceride prodrugs and prepare them into nanoemulsions, and improve the oral absorption of docetaxel by simulating the characteristics of oral absorption of natural triglyceride lymphatic transport. However, nanoemulsions have the disadvantages of storage instability, cumbersome preparation, poor batch reproducibility, etc., and a large amount of co-emulsifiers such as sodium deoxycholate may be added in the preparation of nanoemulsions. Multiple administrations may cause serious gastrointestinal side effects. Therefore, it is very necessary to design a safe and simple oral dosage form of triglyceride prodrugs. Summary of the invention

[0005] In view of the problem of poor oral absorption of docetaxel, the present invention first provides a docetaxel-stearate triglyceride prodrug based on the natural triglyceride lymphatic transport mechanism, and prepares the prodrug into a lipid preparation of docetaxel-stearate triglyceride prodrug. The preparation process is simple, repeatable, easy to industrialize, and uniform and stable. The prodrug lipid preparation can promote the lymphatic transport of poorly soluble drugs and improve their oral bioavailability. On this basis, a reduction-sensitive linker is introduced to promote the oral absorption of anti-tumor drugs while enabling them to be specifically released at the target site, thereby improving the efficacy and reducing toxicity.

[0006] The first object of the present invention is to provide docetaxel-stearate triglyceride prodrug or its geometric isomers, pharmaceutically acceptable salts, hydrates, solvates:

[0007]

[0008] The second object of the present invention is to provide a method for preparing the above compound. The specific preparation method is as follows:

[0009] (a) Synthesis of 1,3-distearin: A certain amount of stearic acid was dissolved in dichloromethane and reacted with 1,3-dihydroxyacetone to form an ester under the catalysis of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDCI) and 4-dimethylaminopyridine (DMAP), and then hydrogenated with sodium borohydride to obtain 1,3-distearin.

[0010]

[0011] (b) Dissolving dithiodibutyric acid in an appropriate amount of acetic anhydride, stirring at room temperature to form dithiodibutyric anhydride, reacting with 1,3-distearin in the presence of EDCI and DMAP to obtain an intermediate product, and esterifying with docetaxel under the catalytic conditions of DMAP and EDCI to obtain the target prodrug.

[0012]

[0013] The third object of the present invention is to provide a simple and safe pharmaceutical composition containing docetaxel-stearate triglyceride prodrug. The pharmaceutical composition is a lipid preparation of docetaxel-stearate triglyceride prodrug, comprising docetaxel-stearate triglyceride prodrug and excipients. The excipients include phospholipid emulsifiers, short-chain alcohols or ethers emulsifiers, and liquid oil phases. By weight percentage, the liquid oil phase accounts for 30%-85% of the lipid preparation, the phospholipid emulsifier accounts for 9%-45% of the lipid preparation, and the short-chain alcohols or ethers emulsifiers account for 5%-25% of the lipid preparation; and the docetaxel-stearate triglyceride prodrug accounts for 1-10% of the total weight of the excipients.

[0014] Furthermore, in the pharmaceutical composition of the present invention, the liquid oil phase accounts for 57%-85% of the lipid preparation, the phospholipid emulsifier accounts for 9%-33% of the lipid preparation, and the short-chain alcohol or ether emulsifier accounts for 5%-10% of the lipid preparation by weight percentage;

[0015] Preferably, in the pharmaceutical composition of the present invention, by weight percentage, the liquid oil phase accounts for 57%-75% of the lipid preparation, the phospholipid emulsifier accounts for 20%-33% of the lipid preparation, and the short-chain alcohol or ether emulsifier accounts for 5%-10% of the lipid preparation.

[0016] Further, the docetaxel-stearate triglyceride prodrug accounts for 2-5% of the total weight of the excipients;

[0017] Preferably, the docetaxel-stearate triglyceride prodrug accounts for 2-4% of the total weight of the excipients.

[0018] Wherein, the phospholipid is egg yolk lecithin, soybean lecithin, synthetic phospholipids such as 1,2-dioctanoyl-sn-glycero-3-phosphocholine, 1,2-didecanoyl-sn-glycero-3-phosphocholine, preferably egg yolk lecithin;

[0019] The liquid oil is selected from long-chain triglycerides, mixed long-chain glycerides, medium-chain triglycerides or a combination thereof, including: one or more of olive oil, almond oil, canola oil, castor oil, coconut oil, corn oil, cottonseed oil, fish oil, palm kernel oil, palm oil, peanut oil, rapeseed oil, safflower oil, sesame oil, shark liver oil, soybean oil, sunflower oil, hydrogenated coconut oil, hydrogenated cottonseed oil, hydrogenated palm oil, hydrogenated soybean oil, partially hydrogenated soybean oil, hydrogenated vegetable oil, Maisine, Peceol, preferably olive oil.

[0020] The short-chain alcohol or ether emulsifier is diethylene glycol monoethyl ether (Transcutol HP), ethanol, propylene glycol, etc., among which Transcutol HP is preferred.

[0021] Furthermore, the weight ratio of the short-chain alcohol or ether emulsifier, the phospholipid emulsifier, and the liquid oil phase is 1:2-4:5-20.

[0022] The lipid preparation of docetaxel-stearate triglyceride prodrug of the present invention is prepared by the following method:

[0023] Docetaxel-stearate triglyceride prodrug, short-chain alcohol or ether emulsifier, phospholipid emulsifier and liquid oil phase are mixed, and the auxiliary materials and the prodrug are uniformly mixed by ultrasound to obtain the lipid preparation of the prodrug.

[0024] Alternatively, the phospholipid emulsifier and the short-chain alcohol or ether emulsifier are dissolved in liquid oil, and ultrasonically mixed to obtain a uniform blank preparation. Docetaxel-stearate triglyceride prodrug is added to the blank preparation, and ultrasonically mixed to completely dissolve the prodrug therein, thereby obtaining a uniform lipid oral preparation.

[0025] When the phospholipid is egg yolk lecithin, the co-emulsifier is diethylene glycol monoethyl ether, the liquid oil is olive oil, and the liquid oil phase accounts for 57%-75% of the lipid preparation, the phospholipid emulsifier accounts for 20%-33% of the lipid preparation, and the short-chain alcohol or ether co-emulsifier accounts for 5%-10% of the lipid preparation, the obtained lipid preparation of docetaxel-stearate triglyceride prodrug has an optimal particle size and particle size distribution, and has better stability, and can significantly improve the oral bioavailability of docetaxel.

[0026] The present invention uniformly mixes liquid oil (especially olive oil), phospholipids and diethylene glycol monoethyl ether for the first time to prepare a blank lipid preparation, and dissolves the synthesized reduction-sensitive docetaxel-stearate triglyceride prodrug into the above preparation. The preparation method of the lipid preparation is simple, and does not require high-temperature heating, probe ultrasound and other processes, and only requires simple stirring or ultrasonic dissolution, the process is simple, easy to industrialize, and has strong repeatability; the obtained lipid preparation is in a uniform oil solution state, and relative to the thermodynamically unstable state of the nano preparation, the lipid preparation provided by the present invention is more stable and easy to store; the use of sodium deoxycholate is avoided, and the excipients used in the lipid preparation provided by the present invention are all FDA-approved pharmaceutical excipients, and will not cause gastrointestinal toxicity after oral administration; the drug loading amount is greatly improved, and the preparation and patient compliance will be greatly improved; in addition, the lipid preparation is highly suitable for the designed docetaxel-stearate triglyceride prodrug, and significantly improves the oral bioavailability of docetaxel.

[0027] The advantages of the present invention are:

[0028] 1. Based on the absorption mechanism of natural triglycerides, the present invention designs docetaxel-stearate triglyceride prodrug, which promotes the lymphatic transport of docetaxel, avoids the first-pass effect, and thus improves its oral absorption.

[0029] 2. The present invention connects the triglyceride backbone and the poorly soluble drug with a reduction-sensitive disulfide bond bridge, which promotes oral absorption of the drug and enables the drug to be specifically released at the target site, thereby increasing efficacy and reducing toxicity.

[0030] 3. The lipid oral preparation prepared by the present invention has a simple preparation process, is easy to industrialize, is uniform and stable, and, compared with the nanoemulsion in the prior art, avoids the use of sodium deoxycholate, simplifies the preparation process, and does not require a heating process or probe ultrasound. BRIEF DESCRIPTION OF THE DRAWINGS

[0031] Figure 1 Docetaxel-stearate triglyceride prodrug 1 H-NMR spectrum.

[0032] Figure 2 This is the high-resolution mass spectrum of docetaxel-stearate triglyceride prodrug.

[0033] Figure 3 The storage stability of the docetaxel-stearate triglyceride prodrug nanoemulsion in Example 2.

[0034] Figure 4 is the solubility of docetaxel-stearate triglyceride prodrug in different excipients.

[0035] Figure 5 Pseudo-ternary phase diagram for medium-chain triglyceride-based self-microemulsion screening of docetaxel-stearate triglyceride prodrug.

[0036] Figure 6 The concentration-time curves of medium-chain self-microemulsification and olive oil solutions of docetaxel-stearate triglyceride prodrug in SD rats.

[0037] Figure 7 This is the concentration-time curve of docetaxel-stearate triglyceride prodrug lipid formulation in the pharmacokinetic experiment of SD rats.

[0038] Figure 8 Gastrointestinal toxicity of a lipid formulation of docetaxel-triglyceride prodrug. DETAILED DESCRIPTION

[0039] The present invention is further described by the following examples, but is not limited thereto.

[0040] Example 1 Preparation of docetaxel-stearate triglyceride prodrug (SATG)

[0041] The structure is as follows

[0042]

[0043] 11.12g (40mmol) of stearic acid was dissolved in dichloromethane, and reacted with 1.75g ​​(20mmol) of 1,3-dihydroxyacetone to form an ester under the catalysis of EDCI and DMAP. The reaction solution was concentrated, washed twice with redistilled water, and the water layer was extracted with chloroform. The organic layer was washed once with saturated brine, and the organic layer was dried with anhydrous sodium sulfate, and the solvent was removed by rotary evaporation. Column chromatography separation, separation conditions: n-hexane-ethyl acetate (30:1-15:1). 2.9g (5mmol) of the above product was dissolved in 100ml of a mixed solvent (THF: benzene: water 10:2:1), 0.3g (7.8mmol) of sodium borohydride was added, and the reaction was carried out at 5°C for 30min, 0.9ml of glacial acetic acid was added to stop the reaction, 80ml of chloroform was added to the reaction solution, and the mixture was washed once with redistilled water, once with a 4% sodium bicarbonate solution, and once with saturated brine, and dried with anhydrous sodium sulfate. The solvent was removed by rotary evaporation to obtain 1,3-DG. Dissolve 2.1g (8.8mmol) of 4,4'-dithiodibutyric acid in 9mL of acetic anhydride, react at room temperature for 2h, concentrate the solvent by rotary evaporation and redissolve in an appropriate amount of anhydrous dichloromethane, and add 620mg (1mmol) of 1,3-DG and 100mg of DMAP. Vacuum and react at room temperature for 12 hours under nitrogen protection. Rotary evaporation to remove the solvent, column chromatography to separate the intermediate product (conditions are n-hexane: ethyl acetate = 15:1-8:1). Dissolve 680mg (0.83mmol) of the intermediate product in an appropriate amount of anhydrous dichloromethane, add 175mg (0.9mmol) of EDCI, 45mg (0.36mmol) of DMAP, and 803mg (1mmol) of docetaxel, vacuum and react at room temperature for 48 hours under nitrogen protection. Rotary evaporation to remove the solvent, and preparative liquid phase separation to obtain the final product (preparative liquid phase conditions are pure acetonitrile).

[0044] Nuclear magnetic resonance imaging 1 H-NMR hydrogen spectrum was used to determine the structure of the prodrug in Example 1. The solvent used was CDCl3. The results are as follows Figure 1 . Corresponding characteristic peak analysis: docetaxel characteristic peak: 1H NMR (400MHz, CDCl3) δ8.12 (d, J = 7.7Hz, 2H), 7.60 (d, J = 7.2Hz, 1H), 7.50 (t, J = 7.6Hz, 2H), 7.41–7.23 (m, 5H), 6.25 (t, J = 8.0Hz, 1H); 1,3-distearic acid glyceryl ester skeleton characteristic peak: 1.27 (d, J = 9.9Hz, 56H), 0.88 (t, J = 6.7Hz, 6H);

[0045] The molecular weight of the prodrug in Example 1 was determined by high resolution mass spectrometry. Figure 2 Shown: ESI-HRMS: Calcd.For C 90 H 139 NO21 S2Cl[M+Cl - ]1668.9022found 1696.8909. The specific synthetic route of the above prodrug is as follows:

[0046]

[0047]

[0048] Example 2 Stability of SATG Nanoemulsion

[0049] 12 mg of docetaxel-stearate triglyceride prodrug was dissolved in 400 mg of olive oil and preheated to 60°C. 120 mg of egg yolk lecithin and 40 mg of sodium deoxycholate were weighed and dissolved in 4 ml of deionized water and preheated to 60°C. The oil phase was slowly dripped into the stirred aqueous phase and stirred for 3 minutes to form colostrum. The colostrum was ultrasonicated in an ice bath probe for 10 minutes with an ultrasonic power of 500 W to prepare docetaxel-stearate triglyceride nanoemulsion. The particle size of the emulsion was measured on days 1, 2, 5, 15, and 30. It was found that the particle size of the emulsion increased significantly over time, and even stratification occurred in the end, exposing the defect of the instability of the emulsion. The results are as follows: Figure 3 As shown, the drug loading of SATG nanoemulsion is only 0.258%, which shows that the preparation of docetaxel-stearate triglyceride into nanoemulsion not only has a low drug loading but also poor stability.

[0050] Example 3: Formulation Optimization of Docetaxel-Triglyceride Prodrug Lipid Formulation

[0051] The oral absorption of triglyceride-like prodrugs is highly dependent on lipid digestion. Therefore, the best carrier for SATG is a lipid-based drug delivery system (LBDDS). Among the various types of LBDDS, self-microemulsified drug delivery system (SMEDDS) has unique advantages, including high drug loading and easy to scale up production.

[0052] 3.1 Preliminary screening and preparation of SATG self-microemulsions based on medium-chain triglycerides (MCT) and long-chain triglycerides (LCT)

[0053] Firstly, the solubility of SATG in different oil-soluble excipients was determined, such as Figure 4 As shown in the figure, it was found that SATG had the highest solubility in tricaprylin (MCT), monolinolein (Maisine35-1), and diethylene glycol monoethyl ether (Trancutol HP), so they were used as oil phase, emulsifier, and co-emulsifier. A pseudo-ternary phase diagram was established to find the most suitable ratio of the three items, such as Figure 5As shown, the preparation prepared at the ratio in the dotted area can form a self-microemulsion by simulating the stirring in the stomach. After further optimization, it was found that when MCT / Maisine 35-1 / Transcutol HP=68.2 / 16.1 / 15.7 (weight ratio), the emulsion particle size formed was smaller and more uniform.

[0054] Based on solubility and particle size of emulsion formation, a medium-chain triglyceride-based self-microemulsification drug delivery system was developed after PDI screening. Based on the solubility of the prodrug in each excipient, a docetaxel-stearate triglyceride lipid formulation was prepared with a total weight ratio of docetaxel-stearate triglyceride prodrug to the above excipients of 1:25.

[0055] The in vivo pharmacokinetic experiment was carried out according to the above formula, and a SATG olive oil solution (long-chain triglyceride LCT) group was set up for comparison. Figure 6 As shown in the figure, compared with the SMEDDS based on MCT (medium chain triglycerides), the AUC (0-8h) of the SATG olive oil (LCT) solution group increased significantly. It can be seen that although the emulsion particle size formed by the formulation designed according to the pseudo-ternary phase diagram is smaller and more uniform in medicine, it does not show good in vivo pharmacokinetic properties. The LCT-based formulation showed better in vivo pharmacokinetic properties than the MCT-based formulation, so the long-chain triglyceride LCT is more suitable for the design of SATG self-microemulsion.

[0056] The results showed that the main mechanism of SATG is to mimic the oral absorption process of long-chain triglycerides, which is digested, absorbed into intestinal cells, and re-esterified and assembled into chylomicrons, transported to the lymphatic system and then circulated. Co-administration with LCT, rather than MCT, can enhance the solubility of SAMG (the product obtained after digestion of SATG by intestinal fluid) in the intestine, thereby improving the efficiency of oral absorption.

[0057] Therefore, the present invention selects long-chain triglyceride LCT as the oil phase for further experiments.

[0058] 3.2 Prescription screening of SATG lipid preparations based on long-chain triglycerides

[0059] 3.2.1 Selection of oil phase

[0060] Based on the above results, in order to ensure the drug loading capacity of the lipid preparation, the solubility of SATG in several LCT-based vegetable oils was measured. The results are shown in Table 1.

[0061] Table 1. Solubility of SATG in several vegetable oils

[0062]

[0063] Based on the solubility results, olive oil was selected as the oil phase of the SATG lipid formulation.

[0064] 3.2.2 Selection of emulsifier

[0065] Phospholipids can help improve the oral absorption of triglyceride-like prodrugs, so egg yolk lecithin was initially selected as an emulsifier to screen for co-emulsifiers. The emulsifier and co-emulsifier were dissolved in the oil phase, with the mass ratio of oil phase: emulsifier: co-emulsifier being 65:25:10, and the mixture was uniformly mixed by ultrasound to obtain a uniform blank preparation.

[0066] 200 mg of SATG prodrug was added to 1 g of the above blank preparation to measure the equilibrium solubility of SATG prodrug in the blank preparation.

[0067] Another 30 mg of SATG prodrug was added to 1 g of the blank preparation, and the prodrug was completely dissolved therein by ultrasound to obtain a uniform lipid oral preparation. 500 mg of each drug-containing preparation was added to 5 mL of simulated gastric fluid and stirred for 5 min to simulate gastric motility. The particle size and distribution were measured. The results of different emulsifiers are shown in Table 2.

[0068] Table 2. Properties of SATG lipid preparations prepared with different emulsifiers

[0069]

[0070] The results showed that the performance of the prepared lipid preparations was different when different emulsifiers were added. When the emulsifier was Transcutol HP, the solubility of the prodrug was the highest and the optimal drug loading could be obtained. The prepared SATG prodrug lipid preparation had the optimal drug loading, particle size and distribution.

[0071] 3.2.3 Selection of phospholipids

[0072] Using olive oil as the oil phase and Transcutol HP as the emulsifier, the types of phospholipids were screened, and egg yolk lecithin, soybean lecithin, 1,2-dioctanoyl-sn-glycero-3-phosphocholine, and 1,2-didecanoyl-sn-glycero-3-phosphocholine were selected. SATG lipid preparations were prepared according to the method in 3.2.2, and the equilibrium solubility of SATG prodrug in the blank preparation was determined. 500 mg of each drug-containing preparation was added to 5 mL of simulated gastric fluid and stirred for 5 minutes to simulate gastric motility. The particle size and distribution were determined. There was no significant difference in the various indicators of lipid preparations prepared with different phospholipids, and egg yolk lecithin was relatively better.

[0073] Example 4 Preparation of SATG prodrug lipid formulation

[0074] SATG prodrug lipid preparations were prepared according to the method of 3.2.2 in Example 3 to obtain prodrug lipid preparations of different formulations, as shown in Table 3. 500 mg of the lipid preparation was added to 5 mL of simulated gastric juice, stirred for 5 min to simulate gastric peristalsis, and the particle size and distribution were measured. Another 1 mL of the above lipid preparation and simulated gastric juice mixture was mixed with 5 mL of pancreatic juice-bile, incubated for 2 h, centrifuged at low temperature (10000 rpm, 10 min), and the middle layer liquid was taken to measure the particle size after digestion and the proportion of SATG in the middle layer. The results are shown in Table 4:

[0075] Table 3. SATG lipid preparations of different formulations

[0076]

[0077]

[0078] Table 4. Properties of SATG lipid preparations of different formulations

[0079]

[0080] As shown in Table 4, when the proportion of olive oil is less than 50% (formulations 1-3), egg yolk lecithin and Transcutol HP account for a high proportion, and the solubility of SATG is high, so the drug loading is high. In addition, due to the strong surface activity of the emulsifier and the co-emulsifier, after mixing and stirring with simulated gastric juice, the particles obtained are small and uniform in size, which is similar to the in vitro results of the self-microemulsification preparation based on MCT. However, after mixing with pancreatic juice-bile to simulate digestion, the particle size of the aqueous phase of the intermediate layer is larger, and the proportion of SATG prodrug in the intermediate layer is less than 50%. For triglyceride-like lipid preparations, the initial in vitro particle size cannot accurately predict the oral absorption in vivo, and the intermediate aqueous phase after simulated digestion is equivalent to the dissolved phase of the intestinal fluid in vivo. The smaller the particle size, the greater the proportion of the prodrug in the intermediate aqueous phase, which may be beneficial to in vivo absorption. When the proportion of olive oil exceeds 80% and the proportion of egg yolk lecithin is less than 20%, although the results of in vivo simulation are still within an acceptable range, the initial simulated gastric fluid emulsification particle size is greater than 300nm, which may delay the subsequent intestinal digestion process and thus affect oral absorption; when TrancutolHP is less than 5%, olive oil and egg yolk lecithin are incompatible and cannot form a uniform lipid preparation.

[0081] When the content of olive oil is 30%-85%, the content of egg yolk lecithin is 9%-45%, and the content of TrancutolHP is 5%-25%, a uniform and stable lipid preparation can be formed, and the particle size of the lipid preparation is less than 400nm.

[0082] When the content of olive oil is 57%-85%, the content of egg yolk lecithin is 9%-33%, and the content of TrancutolHP is 5%-10%, the particle size of the prepared lipid preparation is less than 400nm, and SATG accounts for a large proportion in the middle layer, which is beneficial for absorption in the body.

[0083] When the content of olive oil is 57%-75%, the content of egg yolk lecithin is 20%-33%, and the content of TrancutolHP is 5%-10%, the particle size of the prepared lipid preparation is less than 300nm and is most conducive to in vivo absorption.

[0084] The lipid preparations of each formulation had no stratification and precipitation at 15 days, and had good stability.

[0085] Example 5 Preparation of SATG prodrug lipid formulation

[0086] Weigh 300 mg of egg yolk lecithin and dissolve it in 1 g of olive oil. At the same time, add 100 mg of diethylene glycol monoethyl ether and mix it evenly by ultrasound to obtain a uniform blank preparation. Accurately weigh 40 mg of the prodrug described in Example 1, add 1 g of the blank preparation, and ultrasound to completely dissolve the prodrug therein to obtain a uniform SATG prodrug lipid preparation. Accurately weigh a certain amount of docetaxel parent drug and add it to the above blank preparation to obtain a docetaxel lipid preparation.

[0087] Example 6 Pharmacokinetics of docetaxel-triglyceride SATG prodrug lipid formulation

[0088] Using SD rats as a model, the lipid formulation containing docetaxel-triglyceride prodrug (SATGLP) and the lipid formulation containing docetaxel (DTX LP) of Example 5 were orally administered, and the relative dose of docetaxel was 10 mg / kg. Blood was collected from the orbits at regular intervals to determine the concentration of docetaxel parent drug in plasma. The drug-time curves were drawn and the corresponding pharmacokinetic parameters were calculated according to the determined concentrations (Table 5). In order to calculate the absolute bioavailability, docetaxel solution was intravenously administered at a dose of 5 mg / kg, and the docetaxel content in plasma was determined. Figure 7 As shown in Table 5, relative to docetaxel lipid formulation, the area under the drug-time curve (AUC 0-24 ) was significantly improved relative to the parent drug lipid preparation. The oral bioavailability of the prodrug was calculated from the data of intravenous docetaxel, and the absolute bioavailability of the prodrug group reached 17.8%. In addition, the total amount of docetaxel in the blood of the SATG prodrug lipid preparation group was determined by fragmentation, and the absolute bioavailability of the total amount of docetaxel in the prodrug group was calculated to be 121%, which significantly improved the oral absorption of docetaxel.

[0089] Table 5. Main pharmacokinetic parameters of docetaxel triglyceride prodrug

[0090]

[0091] Example 7 Gastrointestinal toxicity of docetaxel-triglyceride SATG prodrug lipid formulation

[0092] The cytotoxicity of SATG prodrug lipid preparations on human cloned colon adenocarcinoma cells (CaCo-2) was investigated by MTT method. Cells were seeded into 96-well plates at a density of 1000 cells / well and incubated in an incubator for 24 hours to adhere to the wall. After the cells adhered to the wall, a series of concentrations of paclitaxel solution, the SATG nanoemulsion prepared in Example 2, and the SATG prodrug lipid preparation prepared in Example 5 were added. After 12 hours of drug addition, the cell survival rate was determined by MTT method.

[0093] The results are as follows Figure 8 As shown, the cytotoxicity of SATG prodrug lipid preparations to human cloned colon adenocarcinoma cells is weaker than that of paclitaxel solution and nanoemulsion, indicating that the prodrug lipid preparation has no obvious damage to gastrointestinal cells and the excipients and proportions used have no gastrointestinal toxicity.

Claims

1. A lipid formulation of docetaxel-stearate triglyceride prodrug, characterized in that: It comprises docetaxel-stearate triglyceride prodrug and excipients, wherein the excipients are composed of a phospholipid emulsifier, a short-chain alcohol or ether emulsifier and a liquid oil phase. By weight percentage, the liquid oil phase accounts for 57%-85% of the lipid preparation, the phospholipid emulsifier accounts for 9%-33% of the lipid preparation, and the short-chain alcohol or ether emulsifier accounts for 5%-10% of the lipid preparation; the docetaxel-stearate triglyceride prodrug accounts for 1-10% of the total weight of the excipients; the short-chain alcohol or ether emulsifier is diethylene glycol monoethyl ether, and the liquid oil phase is a long-chain triglyceride; the phospholipid emulsifier is egg yolk lecithin, soybean lecithin, 1,2-dioctanoyl-sn-glycero-3-phosphocholine or 1,2-didecanoyl-sn-glycero-3-phosphocholine; the long-chain triglyceride is olive oil; The lipid preparation of docetaxel-stearate triglyceride prodrug is prepared by the following method: The docetaxel-stearate triglyceride prodrug, a short-chain alcohol or ether emulsifier, a phospholipid emulsifier and a liquid oil phase are mixed, and the excipients and the prodrug are uniformly mixed by ultrasonication to obtain the product; Alternatively, a phospholipid emulsifier and a short-chain alcohol or ether emulsifier are dissolved in a liquid oil phase, and ultrasonically mixed to obtain a uniform blank preparation. Docetaxel-stearate triglyceride prodrug is added to the blank preparation, and ultrasonically mixed to obtain a uniform blank preparation. The structure of the docetaxel-stearate triglyceride prodrug is as follows: 。 2. The lipid preparation of docetaxel-stearate triglyceride prodrug according to claim 1, characterized in that: Liquid oil phase accounts for 57%-75% of lipid preparations, phospholipid emulsifiers account for 20%-33% of lipid preparations, and short-chain alcohol or ether emulsifiers account for 5%-10% of lipid preparations.

3. The lipid preparation of docetaxel-stearate triglyceride prodrug according to claim 1, characterized in that: The weight ratio of the short-chain alcohol or ether emulsifier, the phospholipid emulsifier and the liquid oil phase is 1:2-4:5-20.

4. The lipid formulation of docetaxel-stearate triglyceride prodrug according to any one of claims 1 to 3, characterized in that: The docetaxel-stearate triglyceride prodrug accounts for 2-5% of the total weight of the excipients.

5. The lipid formulation of docetaxel-stearate triglyceride prodrug according to any one of claims 1 to 3, characterized in that: The docetaxel-stearate triglyceride prodrug accounts for 2-4% of the total weight of the excipients.

6. The lipid formulation of docetaxel-stearate triglyceride prodrug according to claim 1, characterized in that: The lipid preparation is composed of egg yolk lecithin, docetaxel-stearate triglyceride prodrug, olive oil and diethylene glycol monoethyl ether, wherein olive oil accounts for 57%-75% of the lipid preparation, egg yolk lecithin accounts for 20%-33% of the lipid preparation, diethylene glycol monoethyl ether accounts for 5%-10% of the lipid preparation, and docetaxel-stearate triglyceride prodrug accounts for 2-4% of the total weight of the excipients.

7. The lipid formulation of docetaxel-stearate triglyceride prodrug according to claim 1, characterized in that: The preparation of the docetaxel-stearate triglyceride prodrug comprises the following steps: (a) Synthesis of 1,3-distearic acid glyceryl ester: Stearic acid is dissolved in dichloromethane, and reacted with 1,3-dihydroxyacetone to form an ester under the catalysis of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride and 4-dimethylaminopyridine, and then hydrogenated with sodium borohydride to obtain 1,3-distearic acid glyceryl ester; (b) Dissolve dithiodibutyric acid in acetic anhydride, stir at room temperature to form dithiodibutyric anhydride, react with 1,3-distearic acid glycerol in the presence of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride and 4-dimethylaminopyridine to obtain an intermediate product, and then react with docetaxel to obtain an intermediate product.

8. Use of the lipid preparation of docetaxel-stearate triglyceride prodrug according to any one of claims 1 to 7 in the preparation of anti-tumor drugs.

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