Preparation of Docetaxel-Triolein Prodrug and Lipid Preparation

Abstract

The present invention belongs to the field of pharmaceutical technology, and relates to docetaxel triglyceride prodrugs and their lipid preparations, as well as docetaxel-triglyceride prodrugs mediated by lymphatic transport and their lipid preparations and their application in the preparation of oral chemotherapy drugs. The present invention provides a lipid preparation of docetaxel-triglyceride prodrug, which comprises a docetaxel-triglyceride prodrug and excipients. The excipients include phospholipid emulsifiers, short-chain alcohol or ether co-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 10%-45% of the lipid preparation, and the short-chain alcohol or ether co-emulsifier accounts for 5%-25% of the lipid preparation; the docetaxel-triglyceride prodrug accounts for 1%-10% of the total weight of the excipients. The docetaxel-triglyceride prodrug of the present invention promotes the lymphatic transport of docetaxel, avoids the first-pass effect, and thus improves its oral absorption.

Description

Technical Field

[0001] The present invention belongs to the field of pharmaceutical technology, and relates to docetaxel-triglyceride prodrug and its lipid preparation, specifically to docetaxel-triglyceride prodrug with lymph-mediated transport and its lipid preparation and the application in the preparation of oral chemotherapy drugs. Background Art

[0002] Nowadays, the global cancer burden increases year by year. Docetaxel (DTX), as a first-line broad-spectrum anti-cancer chemotherapy drug, is widely used in the clinical treatment of various tumors. Docetaxel is clinically administered by intravenous injection. However, the commercially available intravenous injection solution uses Tween-80 and ethanol for solubilization, bringing excipient-related toxic side effects and limiting its clinical application. Oral chemotherapy has the advantages of high patient compliance, convenient administration, and low treatment cost. However, due to the low solubility of docetaxel, P-glycoprotein efflux, and severe first-pass effect, its oral bioavailability is extremely low, and there is currently no marketed oral preparation of docetaxel. Therefore, the development of docetaxel oral preparations 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 enhance lymphatic transport is to design triglyceride-like prodrugs of drugs. 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 hardly hydrolyzed. Replacing the fatty acid at position 2 with a drug enables the prodrug to mimic the digestion process of triglycerides in the intestine, enter the intestinal epithelial cells in the form of 2-monoacylglycerol prodrugs for re-esterification, participate in the assembly of lipoproteins, and thus promote lymphatic transport. Many researchers have made examples of triglyceride-like prodrugs. However, the structures taught by these examples are almost ineffective in improving drug oral administration. An important reason is the lack of a cleavable linker chain, resulting in the non-release of the parent drug. In CN106715456A, the inventor introduced a self-eliminating linker chain to promote the systemic release of the parent drug testosterone. However, for docetaxel, a chemotherapy drug, a large amount of systemic exposure will cause serious side effects. Therefore, this linker chain is not instructive for the design of docetaxel oral delivery systems.

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

[0005] Aiming at the problem of poor oral absorption of docetaxel, based on the natural lymphatic transport mechanism of triglycerides, the present invention first provides a docetaxel-triolein prodrug, and prepares a lipid preparation of the docetaxel-triolein prodrug. The preparation process is simple, has strong repeatability, is convenient for industrialization, and has uniform and stable properties. This prodrug lipid preparation can promote the lymphatic transport of poorly soluble drugs, improve their oral bioavailability, and on this basis, introduce a reduction-sensitive linker, which can not only promote the oral absorption of anti-tumor drugs, but also enable them to be specifically released at the target site, improve the curative effect and reduce the toxicity.

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

[0007]

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

[0009] (a) Synthesis of 1,3-diolein: Oleic acid is dissolved in dichloromethane, and an esterification reaction occurs with 1,3-dihydroxyacetone under the catalysis of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDCI) and 4-dimethylaminopyridine (DMAP), and then a hydrogenation reaction with sodium borohydride gives 1,3-diolein.

[0010]

[0011] (b) Dissolve dithiodibutyric acid in an appropriate amount of acetic anhydride, stir at room temperature to form dithiodibutyric anhydride, react with 1,3-diolein in the presence of EDCI and DMAP to obtain an intermediate product, and then carry out an esterification reaction with docetaxel under the catalysis of DMAP and EDCI to obtain the target prodrug.

[0012] The third object of the present invention is to provide a simple and safe pharmaceutical composition containing a docetaxel-triolein prodrug. The pharmaceutical composition is a lipid preparation of the docetaxel-triolein prodrug, which contains the docetaxel-triolein prodrug and excipients. The excipients include phospholipid emulsifiers, short-chain alcohol or ether co-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 10%-45% of the lipid preparation, and the short-chain alcohol or ether co-emulsifier accounts for 5%-25% of the lipid preparation; the docetaxel-triolein prodrug accounts for 1-10% of the total weight of the excipients.

[0013] Furthermore, in the pharmaceutical composition of the present invention, by weight percentage, the liquid oil as the oil phase accounts for 55%-85% of the lipid preparation, the phospholipid emulsifier accounts for 10%-35% of the lipid preparation, and the short-chain alcohol or ether co-emulsifier accounts for 5%-10% of the lipid preparation.

[0014] Preferably, in the pharmaceutical composition of the present invention, by weight percentage, the liquid oil as the oil phase accounts for 55%-80% of the lipid preparation, the phospholipid emulsifier accounts for 15%-35% of the lipid preparation, and the short-chain alcohol or ether co-emulsifier accounts for 5%-10% of the lipid preparation.

[0015] Furthermore, the docetaxel-triglyceride oleate prodrug accounts for 2-4% of the total weight of the excipients.

[0016] Preferably, the docetaxel-triglyceride oleate prodrug accounts for 3-4% of the total weight of the excipients.

[0017] Among them, 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, and preferably egg yolk lecithin;

[0018] The liquid oil is selected from long-chain triglycerides, mixed long-chain glycerides, medium-chain triglycerides or combinations thereof, including: 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, tricaprylin, Maisine, Peceol, and one or more of them, and preferably olive oil.

[0019] The short-chain alcohol or ether co-emulsifier is diethylene glycol monoethyl ether (Transcutol HP), ethanol, propylene glycol, etc., and preferably Transcutol HP.

[0020] Even further, the weight ratio of the short-chain alcohol or ether co-emulsifier, the phospholipid emulsifier, and the liquid oil as the oil phase is: 1:2-4:5-20.

[0021] The lipid preparation of the docetaxel-triglyceride oleate prodrug of the present invention is prepared by the following method:

[0022] Mix the docetaxel-triglyceride oleate prodrug, the short-chain alcohol or ether co-emulsifier, the phospholipid emulsifier, and the liquid oil as the oil phase, and ultrasonically mix the excipients and the prodrug uniformly to obtain the lipid preparation of the prodrug.

[0023] Alternatively, dissolve phospholipid emulsifiers and short-chain alcohol or ether co-emulsifiers in liquid oil, and ultrasonically mix them evenly to obtain a uniform blank preparation. Take the docetaxel-triglyceride prodrug, add it to the above blank preparation, and ultrasonically dissolve the prodrug completely in it to obtain a uniform lipid oral preparation.

[0024] When the phospholipid emulsifier is egg yolk lecithin, the short-chain alcohol or ether co-emulsifier is diethylene glycol monoethyl ether, and the liquid oil phase of the liquid oil is olive oil, and the liquid oil phase of the liquid oil accounts for 55%-80% of the lipid preparation, the phospholipid emulsifier accounts for 15%-35% 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 the docetaxel-triglyceride prodrug has the best particle size and particle size distribution, has the best stability, and can significantly improve the oral bioavailability of docetaxel.

[0025] For the first time, the present invention uniformly mixes liquid oil (especially olive oil), phospholipids and diethylene glycol monoethyl ether to prepare a blank lipid preparation, and dissolves the synthesized reduction-sensitive docetaxel-triglyceride prodrug into the above preparation. The preparation method of the lipid preparation is simple, without high-temperature heating, probe ultrasonic and other processes, only simple stirring or ultrasonic dissolution is required, the process is simple, convenient for industrialization, and has strong repeatability; the obtained lipid preparation is in a uniform oil solution state. Compared with the thermodynamically unstable state of the nano-preparation, the lipid preparation provided by the present invention is more stable and convenient for storage; 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 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-triglyceride prodrug, and significantly improves the oral bioavailability of docetaxel.

[0026] The advantages of the present invention are as follows:

[0027] 1. Based on the mechanism of natural triglyceride absorption, the present invention designs a docetaxel-triglyceride prodrug, which promotes the lymphatic transport of docetaxel, avoids the first-pass effect, and thus improves its oral absorption.

[0028] 2. The present invention uses a reduction-sensitive disulfide bond linker to connect the triglyceride backbone and the poorly soluble drug. While promoting the oral absorption of the drug, the drug can be specifically released at the target site, enhancing the efficacy and reducing the toxicity.

[0029] 3. The lipid oral preparation prepared by the present invention has a simple preparation process, is easy for industrialization, is uniform and stable. Compared with the nanoemulsion in the prior art, the use of sodium deoxycholate is avoided, the preparation process is simplified, and there is no heating process and probe ultrasonic. Description of the Drawings

[0030] Figure 1 Of the docetaxel-triglyceride prodrug 1 1H-NMR spectrum.

[0031] Figure 2 High-resolution mass spectrometry of docetaxel-triglyceride prodrug.

[0032] Figure 3 Particle size stability of the docetaxel-triglyceride prodrug nanoemulsion prepared in Example 2.

[0033] Figure 4 Solubility of docetaxel-triglyceride prodrug in different excipients.

[0034] Figure 5 Pseudo-ternary phase diagram of self-microemulsion screening of docetaxel-triglyceride prodrug based on medium-chain triglycerides.

[0035] Figure 6 Plasma concentration-time curves of self-microemulsion of docetaxel-triglyceride prodrug based on medium-chain triglycerides and olive oil solution in SD rats.

[0036] Figure 7 Plasma concentration-time curves of docetaxel-triglyceride prodrug lipid formulation combined with sodium deoxycholate in SD rats.

[0037] Figure 8 Plasma concentration-time curves of docetaxel-triglyceride prodrug lipid formulation with / without sodium deoxycholate (bile salt) in SD rats.

[0038] Figure 9 Plasma concentration-time curves of docetaxel-triglyceride prodrug lipid formulation in beagle dogs during pharmacokinetic experiments.

[0039] Figure 10 Gastrointestinal toxicity of docetaxel-triglyceride prodrug lipid formulation.

[0040] Figure 11 In vivo oral anti-tumor experimental diagram of docetaxel-triglyceride prodrug lipid formulation.

[0041] A: Tumor growth curve B: Mouse body weight change diagram C: Tumor physical diagram D: Tumor-bearing rate. Detailed implementation mode

[0042] The present invention is further described by the following examples, but is not limited thereto.

[0043] Example 1 Preparation of docetaxel-triglyceride prodrug (OATG)

[0044] The structure is as follows:

[0045] Dissolve 11.28 g (40 mmol) of oleic acid in dichloromethane, and carry out an esterification reaction with 1.75 g (20 mmol) of 1,3-dihydroxyacetone overnight under the catalysis of EDCI and DMAP. Concentrate the reaction solution, wash it twice with redistilled water, extract the aqueous layer with chloroform, wash the organic layer once with saturated brine, dry the organic layer with anhydrous sodium sulfate, and remove the solvent by rotary evaporation. Separate by column chromatography, and the separation conditions are n-hexane-ethyl acetate (30:1 - 15:1). Dissolve 2.9 g (5 mmol) of the above product in 100 ml of a mixed solvent (THF:benzene:water 10:2:1), add 0.3 g (7.8 mmol) of sodium borohydride, react at 5 °C for 30 min, add 0.9 ml of glacial acetic acid to stop the reaction, add 80 ml of chloroform to mix with the reaction solution, wash it once with redistilled water, once with 4% sodium bicarbonate solution, once with saturated brine, dry it with anhydrous sodium sulfate, and remove the solvent by rotary evaporation to obtain 1,3-diolein. Dissolve 2.1 g (8.8 mmol) of 4,4'-dithiobutyric acid in 9 mL of acetic anhydride, react at room temperature for 2 h, concentrate the solvent by rotary evaporation and redissolve it in an appropriate amount of anhydrous dichloromethane, and add 620 mg (1 mmol) of 1,3-diolein and 100 mg of DMAP. React under vacuum and nitrogen protection at room temperature for 12 hours. Remove the solvent by rotary evaporation, and separate by column chromatography to obtain an intermediate product (the conditions are n-hexane:ethyl acetate = 15:1 - 8:1). Dissolve 680 mg (0.83 mmol) of the intermediate product in an appropriate amount of anhydrous dichloromethane, add 175 mg (0.9 mmol) of EDCI, 45 mg (0.36 mmol) of DMAP, and 803 mg (1 mmol) of docetaxel, react under vacuum and nitrogen protection at room temperature for 48 hours. Remove the solvent by rotary evaporation, and separate by preparative liquid chromatography to obtain the final product (the preparative liquid chromatography conditions are pure acetonitrile).

[0046] Determine by nuclear magnetic resonance 1 The 1H-NMR hydrogen spectrum is used to determine the structure of the prodrug in Example 1. The selected solvent is CDCl3, and the results are as Figure 1 . Corresponding characteristic peak analysis: Docetaxel characteristic peaks: δ 8.12 (d, J = 7.3 Hz, 2H), 7.61 (t, J = 7.4 Hz, 1H), 7.51 (t, J = 7.8 Hz, 2H), 7.36 (m, 5H), 6.25 (s, 1H); 1,3-Diolein skeleton characteristic peaks: 2.06 – 1.98 (m, 8H), 1.28 (dd, J = 19.6, 7.7 Hz, 40H), 0.88 (t, J = 7.0 Hz, 6H);

[0047] The molecular weight of the prodrug in Example 1 was determined by high-resolution mass spectrometry, and the results are as Figure 2 shown below:

[0048] ESI-HRMS: Calcd. For C 90 H 135 NNaO 21 S2 [M + Na + 1652.8896 found 1652.8860.

[0049] The specific synthesis route of the above prodrug is as follows:

[0050]

[0051]

[0052] Example 2 Stability of OATG nanoemulsion

[0053] 12 mg of docetaxel-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 dropped into the stirred water phase, and stirring was continued for 3 min to form a primary emulsion. The primary emulsion was sonicated in an ice bath with a probe for 10 min at a sonication power of 500 W to obtain the OATG nanoemulsion. The particle size of the emulsion was measured on the 1st, 2nd, 5th, 15th, and 30th days, and it was found that the particle size of the emulsion increased significantly with time, and finally even showed a layering phenomenon, revealing the defect of the instability of the emulsion. The results are as Figure 3 shown. The drug loading of the OATG nanoemulsion was only 0.262%. It can be seen that preparing docetaxel-triglyceride into a nanoemulsion not only has a low drug loading but also poor stability.

[0054] Example 3 Prescription optimization of docetaxel-triglyceride prodrug lipid preparation

[0055] The oral absorption of triglyceride prodrugs highly depends on lipid digestion. Therefore, the optimal carrier for OATG is a lipid-based drug delivery system (LBDDS). Among the various types of LBDDS, the self-microemulsifying drug delivery system (SMEDDS) has unique advantages, including high drug loading and easy scale-up of production.

[0056] 3.1 Preliminary screening and preparation of the OATG self-microemulsion based on medium-chain triglyceride (MCT) and long-chain triglyceride (LCT)

[0057] First, the solubility of OATG in different oil-soluble excipients was determined, such as Figure 4As shown, it was found that OATG had the highest solubility in glyceryl trioctanoate and diethylene glycol monoethyl ether. Therefore, it was used as the oil phase and co-emulsifier. A large number of literature reports have shown that Tween 80 can promote the lymphatic transport of drugs. Therefore, Tween-80 was used as the emulsifier. A pseudo-ternary phase diagram was established to find the most suitable three-component ratio. As Figure 5 shown, the preparation prepared under the ratio enclosed by multiple points in the figure can form self-microemulsions by simulating gastric agitation. After further optimization, it was found that when the ratio of glyceryl trioctanoate / Tween 80 / Transcutol HP = 26.5 / 63 / 10.5 (weight ratio), the formed emulsion had a smaller and more uniform particle size.

[0058] After screening according to solubility, particle size of the formed emulsion, and PDI, a self-microemulsion drug delivery system based on medium-chain triglycerides was developed. According to the solubility of the prodrug in each excipient, a docetaxel-glyceryl oleate prodrug lipid preparation was prepared with a total weight ratio of docetaxel-glyceryl oleate prodrug to the above excipients of 1:20.

[0059] An in vivo pharmacokinetic experiment was conducted on the docetaxel-glyceryl oleate lipid preparation prepared according to the above formulation, and at the same time, a group of olive oil solution of OATG (long-chain triglyceride LCT) was set for comparison.

[0060] The pharmacokinetic results were as Figure 6 shown. Compared with the SMEDDS based on MCT (medium-chain triglycerides), the AUC(0-8h) of the OATG olive oil (LCT) solution group increased significantly (2.6 times). It can be seen that although the emulsion formed pharmacologically according to the formulation designed by the pseudo-ternary phase diagram had a smaller and more uniform particle size, its in vivo pharmacokinetic properties were lower than those of the long-chain triglyceride solution group, that is, the self-microemulsion based on medium-chain triglycerides did not show good in vivo pharmacokinetic properties. And the formulation based on LCT showed better in vivo pharmacokinetic properties than the formulation based on MCT. Therefore, long-chain triglyceride LCT was more suitable for the design of the self-microemulsion of OATG.

[0061] The results showed that the main mechanism of OATG was to mimic the oral absorption process of long-chain triglycerides, which was digested, absorbed into intestinal cells, re-esterified and assembled into chylomicrons, transported to the lymphatic system and then circulated. Co-administration with LCT, rather than MCT, could enhance the solubility of OAMG (the product obtained after digesting OATG by intestinal fluid) in the intestine, thereby improving the oral absorption efficiency.

[0062] Therefore, the present invention selected long-chain triglyceride LCT as the oil phase for further experiments.

[0063] 3.2 Prescription screening of OATG lipid preparation based on long-chain triglycerides

[0064] 3.2.1 Selection of the oil phase

[0065] Based on the above results, to ensure the drug loading of the lipid preparation, the solubility of OATG in several LCT-based vegetable oils was determined, and the results are shown in Table 1.

[0066]

[0067] According to the solubility results, olive oil was selected as the oil phase for the OATG lipid preparation.

[0068] 3.2.2 Selection of co-emulsifier

[0069] Phospholipids can help improve the oral absorption of triglyceride prodrugs. Therefore, egg yolk lecithin was initially selected as the emulsifier to screen the co-emulsifier. The emulsifier and co-emulsifier were dissolved in the oil phase, and the mass ratio of oil phase: emulsifier: co-emulsifier was 65:25:10. Ultrasonic was used to mix them evenly to obtain a homogeneous blank preparation.

[0070] 200 mg of the OATG prodrug was added to 1 g of the above blank preparation, and the equilibrium solubility of the OATG prodrug in the blank preparation was determined.

[0071] In addition, 30 mg of the OATG prodrug was added to 1 g of the above blank preparation, and ultrasonic was used to completely dissolve the prodrug in it to obtain a homogeneous 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 peristalsis, and the particle size and distribution were measured. The results of different co-emulsifiers are shown in Table 2.

[0072]

[0073] The results show that when different co-emulsifiers are added, the properties of the prepared lipid preparations are different. When the co-emulsifier is Transcutol HP, the solubility of the prodrug is the highest, and the best drug loading can be obtained. The prepared OATG prodrug lipid preparation has the best drug loading, particle size and distribution.

[0074] 3.2.3 Selection of phospholipid

[0075] Using olive oil as the oil phase and Transcutol HP as the co-emulsifier, the types of phospholipids were screened. Egg yolk lecithin, soybean lecithin, 1,2-dioctanoyl-sn-glycero-3-phosphocholine, and 1,2-didecanoyl-sn-glycero-3-phosphocholine were selected. The OATG lipid preparation was prepared according to the method of 3.2.2. The equilibrium solubility of the OATG prodrug in the blank preparation was determined, and 500 mg of each drug-containing preparation was added to 5 mL of simulated gastric fluid, and stirred for 5 min to simulate gastric peristalsis, and the particle size and distribution were measured. There were no significant differences in the indexes of the lipid preparations prepared with different phospholipids, and egg yolk lecithin was relatively better.

[0076] Example 4 Preparation of OATG Prodrug Lipid Preparation

[0077] Prepare the OATG prodrug lipid preparation according to the method in 3.2.2 of Example 3 to obtain prodrug lipid preparations with different formulations, as shown in Table 3. Add 500 mg of the lipid preparation to 5 mL of simulated gastric juice, stir for 5 min to simulate gastric peristalsis, measure the particle size and distribution. Take another 1 mL of the mixture of the above lipid preparation and simulated gastric juice and mix it with 5 mL of pancreatic juice-bile, incubate for 2 h, then centrifuge at low temperature (10,000 rpm, 10 min), take the middle layer liquid, measure the particle size after digestion, and the proportion of OATG in the middle layer. The results are shown in Table 4:

[0078]

[0079] As shown in Table 4, when the proportion of olive oil is less than 50% (Formulations 1-3), the proportions of egg yolk lecithin and Transcutol HP are relatively high, and the solubility of OATG is high, so the drug loading is high. In addition, due to the strong surface activity of the emulsifier and co-emulsifier, after mixing and stirring with simulated gastric juice, small and uniform particle sizes are obtained, which is similar to the in vitro results of MCT-based self-microemulsion preparations. However, when mixed with pancreatic juice-bile to simulate digestion, the particle size of the aqueous phase in the middle layer obtained is larger, and the proportion of the OATG prodrug in the middle 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 middle aqueous phase after simulated digestion is equivalent to the dissolution phase of intestinal fluid in vivo. The smaller its particle size and the larger the proportion of the prodrug in the middle aqueous phase, the more likely it is to be beneficial for in vivo absorption. When the proportion of olive oil exceeds 80% and the proportion of egg yolk lecithin is less than 15%, although the in vivo simulation results are still within an acceptable range, the initial emulsification particle size in simulated gastric juice is greater than 300 nm, which may delay the subsequent intestinal digestion process and thus affect oral absorption; when Trancutol HP is less than 5%, olive oil and egg yolk lecithin are incompatible and a homogeneous lipid preparation cannot be formed.

[0080] When the olive oil is at 30%-85%, the egg yolk lecithin is at 10%-45%, and Trancutol HP is at 5%-25%, a homogeneous and stable lipid preparation can be formed, and its particle size is less than 400 nm.

[0081] When the olive oil is at 55%-85%, the egg yolk lecithin is at 10%-35%, and Trancutol HP is at 5%-10%, a homogeneous and stable lipid preparation can be formed, and its particle size is less than 400 nm. The proportion of OATG in the middle layer is large, which is beneficial for in vivo absorption.

[0082] When olive oil is at 55%-80%, egg yolk lecithin is at 15%-35%, and Trancutol HP is at 5%-10%, a homogeneous and stable lipid preparation can be formed, with its particle size less than 300 nm, and it is most conducive to in vivo absorption.

[0083] The lipid preparations of each formulation showed no layering or precipitation at 15 days, indicating good stability.

[0084] Example 5 Optimization of the dosing regimen of the OATG prodrug lipid preparation

[0085] 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 sonicate to mix evenly to obtain a homogeneous blank preparation. Accurately weigh 40 mg of the prodrug described in Example 1 and add it to 1 g of the blank preparation, and sonicate to completely dissolve the prodrug in it to obtain a homogeneous OATG prodrug lipid preparation. Accurately weigh a certain amount of docetaxel mother drug and add it to the above blank preparation to obtain a docetaxel lipid preparation.

[0086] The drug loading of this lipid preparation reached 3.85%, which was 14.6 times higher than that of the nanoemulsion in the prior art. During storage, it remained a homogeneous oily preparation without layering or precipitation.

[0087] Studies have found that sodium deoxycholate can help improve the oral absorption of triglyceride-like prodrugs. According to the prescription shown in Example 5, lipid preparations of OATG and DTX were prepared, and an aqueous solution of sodium deoxycholate was administered to SD rats before the above preparations. Figure 7 It shows the oral plasma distribution after LCT-based SMEDDS administration with sodium deoxycholate solution as a supplement to OATG and DTX. Table 5 gives the corresponding pharmacokinetic parameters. Compared with DTX, the AUC(0-24 h) of the group that simultaneously administered a certain amount of sodium deoxycholate solution with the OATG prodrug increased by 1.86 times. In addition, the Cmax of the OATG group (221.237±82.95 ng mL-1) increased by 6.9 times compared with the DTX group (31.984±13.852 ng mL-1).

[0088]

[0089] However, the use of sodium deoxycholate may cause damage to gastrointestinal function. Therefore, we studied the necessity of sodium deoxycholate in the preparation. The oral plasma curves of SD rats in the OATG group and DTX group after removing sodium deoxycholate from the above preparations are as Figure 8There was no significant difference in the AUC(0–24 h) between the OATG group with sodium deoxycholate (633.916±164.122 µg L h-1) and without sodium deoxycholate (697.1±71.3 µg L h-1). The oral relative bioavailability (Frel) of the OATG group was 247.5%, and the oral absolute bioavailability (Fab) was 37%, in sharp contrast to 15% of the DTX group.

[0090] Example 6 Pharmacokinetics Experiment of Docetaxel-Triglyceride-Oleate Prodrug Lipid Preparation in Beagle Dogs

[0091] Using beagle dogs as models, hard capsules containing docetaxel-triglyceride-oleate prodrug lipid preparation (OATG LP) and hard capsules containing docetaxel lipid preparation (DTX LP) were orally administered at a dose of 3 mg / kg. Blood was collected from the orbital cavity at regular intervals, and the concentration of docetaxel parent drug in plasma was measured. The drug-time curve was plotted and the corresponding pharmacokinetic parameters were calculated based on the measured concentrations (Table 6). To calculate the absolute bioavailability, docetaxel solution was administered intravenously at a dose of 1 mg / kg, and the content of docetaxel in plasma was measured. As Figure 9 shown in Table 6, the area under the drug-time curve (AUC 0-24 ) of the prodrug lipid preparation group was significantly increased compared with that of the docetaxel lipid preparation group. The oral bioavailability of the prodrug was calculated from the data of intravenous docetaxel, and the absolute bioavailability of the prodrug group reached 41.08%.

[0092]

[0093] Example 7 Gastrointestinal Toxicity of Docetaxel-Triglyceride OATG Prodrug Lipid Preparation

[0094] The MTT method was used to investigate the cytotoxicity of the OATG prodrug lipid preparation against human colon adenocarcinoma cells (CaCo-2). Cells were seeded into 96-well plates at a density of 1000 cells / well and incubated in an incubator for 24 h to allow them to adhere. After the cells adhered, a series of concentrations of paclitaxel solution, the OATG nanoemulsion prepared in Example 2, and the OATG prodrug lipid preparation prepared in Example 5 were added. After 12 h of adding the drugs, the cell viability was measured by the MTT method.

[0095] The results are as Figure 10 shown. The cytotoxicity of the OATG prodrug lipid preparation against human colon adenocarcinoma cells was weaker than that of the docetaxel solution and nanoemulsion, indicating that the prodrug lipid preparation had no obvious damage to gastrointestinal cells, and the excipients and ratios used had no gastrointestinal toxicity.

[0096] Example 8 Pharmacodynamic Behavior of Docetaxel-Triglyceride Prodrug Lipid Preparation

[0097] Using a 4T1 orthotopic tumor-bearing Balb / c mouse model, orally administer docetaxel triglyceride lipid formulations, docetaxel lipid formulations, and docetaxel solution. The dosing dose is 10 mg / kg (equivalent dose of docetaxel), and the drug is administered daily. Set up a docetaxel solution intravenous injection group as a positive control, with a dosing dose of 10 mg / kg, and the drug is administered once every two days. Set up a PBS group as a blank control. The results are as Figure 11 shown. The tumor volume in the docetaxel-triglyceride prodrug lipid formulation group is the smallest, showing significant differences from the control groups, and there is no phenomenon of weight loss, indicating its reliable safety and good anti-tumor effect.

Claims

1. Lipid preparation of docetaxel-triglyceride oleate prodrug, characterized in that, It contains docetaxel-triglyceride prodrug and excipients. The excipients include phospholipid emulsifiers, short-chain alcohol or ether co-emulsifiers, and liquid oil as the oil phase. By weight percentage, the liquid oil as the oil phase accounts for 55% - 85% of the lipid preparation, the phospholipid emulsifiers account for 10% - 35% of the lipid preparation, and the short-chain alcohol or ether co-emulsifiers account for 5% - 10% of the lipid preparation; the docetaxel-triglyceride prodrug accounts for 1% - 10% of the total weight of the excipients; The liquid oil as the oil phase is long-chain triglyceride; the short-chain alcohol or ether co-emulsifier is diethylene glycol monoethyl ether; the phospholipid emulsifiers are 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 the docetaxel-triglyceride prodrug is prepared by the following method: Mix the docetaxel-triglyceride prodrug, short-chain alcohol or ether co-emulsifier, phospholipid emulsifiers, and liquid oil as the oil phase, and ultrasonicate to make the excipients and the prodrug uniformly mixed, then it is obtained; Or dissolve the phospholipid emulsifiers and the short-chain alcohol or ether co-emulsifier in the liquid oil as the oil phase, ultrasonicate to make them uniformly mixed to obtain a uniform blank preparation, take the docetaxel-triglyceride prodrug, add it to the above blank preparation, and ultrasonicate to make the prodrug completely dissolve therein, then it is obtained; The docetaxel-triglyceride prodrug or its pharmaceutically acceptable salt: 。 2. The lipid preparation of the docetaxel-triglyceride oleate prodrug according to claim 1, characterized in that, By weight percentage, the liquid oil as the oil phase accounts for 55% - 80% of the lipid preparation, the phospholipid emulsifiers account for 15% - 35% of the lipid preparation, the short-chain alcohol or ether co-emulsifiers account for 5% - 10% of the lipid preparation, and the docetaxel-triglyceride prodrug accounts for 1% - 10% of the total weight of the excipients.

3. The lipid preparation of the docetaxel-triglyceride oleate prodrug according to claim 1 or 2, characterized in that, The docetaxel-triglyceride prodrug accounts for 2% - 4% of the total weight of the excipients.

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

5. The lipid preparation of the docetaxel-triglyceride oleate prodrug according to claim 1, characterized in that, The lipid preparation contains egg yolk lecithin, docetaxel-triglyceride prodrug, olive oil, and diethylene glycol monoethyl ether. Among them, olive oil accounts for 55% - 80% of the lipid preparation, egg yolk lecithin accounts for 15% - 35% of the lipid preparation, diethylene glycol monoethyl ether accounts for 5% - 10% of the lipid preparation, and the docetaxel-triglyceride prodrug accounts for 3% - 4% of the total amount of the excipients.

6. Use of the docetaxel-triglyceride prodrug lipid preparation according to any one of claims 1 - 5 in the preparation of an anti-tumor drug.

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