CN115252802A - Docetaxel-stearic acid triglyceride prodrug and preparation of lipid preparation thereof - Google Patents

Docetaxel-stearic acid triglyceride prodrug and preparation of lipid preparation thereof Download PDF

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CN115252802A
CN115252802A CN202110471401.5A CN202110471401A CN115252802A CN 115252802 A CN115252802 A CN 115252802A CN 202110471401 A CN202110471401 A CN 202110471401A CN 115252802 A CN115252802 A CN 115252802A
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prodrug
docetaxel
oil
preparation
lipid
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田楚彤
杨金诚
冯尧
马宏达
孙进
何仲贵
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Suzhou Yutai Pharmaceutical Technology Co ltd
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    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/54—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic compound
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    • A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
    • A61K31/337—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having four-membered rings, e.g. taxol
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    • A61K47/10—Alcohols; Phenols; Salts thereof, e.g. glycerol; Polyethylene glycols [PEG]; Poloxamers; PEG/POE alkyl ethers
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    • A61K47/24—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing atoms other than carbon, hydrogen, oxygen, halogen, nitrogen or sulfur, e.g. cyclomethicone or phospholipids
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    • C07D305/14—Heterocyclic compounds containing four-membered rings having one oxygen atom as the only ring hetero atoms condensed with carbocyclic rings or ring systems

Abstract

The invention belongs to the technical field of medicines, relates to a docetaxel-triglyceride stearate prodrug and a lipid preparation thereof, and particularly relates to a docetaxel-triglyceride stearate prodrug for lymph-mediated transport, a lipid preparation thereof and application thereof in preparation of oral chemotherapyApplication in medicine. The invention provides a lipid preparation of a docetaxel-triglyceride stearate prodrug, which comprises the docetaxel-triglyceride stearate prodrug and auxiliary materials, wherein the auxiliary materials comprise a phospholipid emulsifier, a short-chain alcohol or ether co-emulsifier and a liquid oil phase, and 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 alcohol or ether co-emulsifier accounts for 5-25% of the lipid preparation according to weight percentage; the docetaxel-stearic acid triglyceride prodrug accounts for 1 to 10 percent of the total weight of the auxiliary materials. The preparation method is simple, convenient for industrialization and strong in repeatability; the obtained lipid preparation is in uniform oil solution state, has good stability, and can improve the curative effect of docetaxel and reduce toxicity.

Description

Docetaxel-stearic acid triglyceride prodrug and preparation of lipid preparation thereof
The technical field is as follows:
the invention belongs to the technical field of medicines, and relates to a docetaxel-triglyceride stearate prodrug and a lipid preparation thereof, in particular to a docetaxel-triglyceride stearate prodrug transported by lymph mediation, a lipid preparation thereof and application thereof in preparation of oral chemotherapy drugs.
The background art comprises the following steps:
nowadays, global cancer burden is increasing year by year, and Docetaxel (DTX) is widely used as a first-line broad-spectrum anticancer chemotherapeutic drug for clinical treatment of various tumors. Docetaxel is clinically administrated by intravenous injection, however, a commercial intravenous injection solution adopts tween-80 and ethanol for assisting dissolution, so that the related toxic and side effects of auxiliary materials are brought, and the clinical application is limited. Oral chemotherapy has the advantages of high patient compliance, convenient administration, low treatment cost and the like, but docetaxel has low solubility, and the oral bioavailability is extremely low due to P-glycoprotein efflux and severe first-pass effect, so that no docetaxel oral preparation is on the market at present. Therefore, the development of docetaxel oral formulations with low first-pass effect and high oral bioavailability is still a hot spot of research.
Oral drug lymphatic transport can avoid the first-pass effect and is an effective way to improve oral absorption. One strategy to increase lymphatic transport is to design triglyceride-like prodrugs of the drug. The fatty acids at the 1-position and the 3-position of the long-chain triglyceride are specifically hydrolyzed by pancreatic lipase, the fatty acid at the 2-position is hardly hydrolyzed, and the fatty acid at the 2-position is replaced by a medicament, so that the prodrug simulates the digestion process of the triglyceride in the intestinal tract, enters intestinal epithelial cells in the form of the 2-monoglyceride prodrug for re-esterification, participates in the assembly of lipoprotein, and further promotes lymphatic transport. Many researchers have made examples of triglyceride-like prodrugs, however, the structures taught by these examples are almost ineffective in enhancing the oral availability of the drug, one important reason being the absence of cleavable linkages, resulting in the non-release of the parent drug. In CN106715456A, the inventors introduced self-elimination of the connecting chain, promoting the systemic release of the parent testosterone. However, for docetaxel, a chemotherapeutic drug, the massive systemic exposure can cause serious side effects, and thus, this link is not instructive in the design of docetaxel oral delivery systems.
In the prior art, a medical worker prepares docetaxel into a triglyceride prodrug and prepares the triglyceride prodrug into a nano-emulsion, and the oral absorption of docetaxel is improved by simulating the characteristic of oral absorption of natural triglyceride lymph transport. However, the nanoemulsion has the disadvantages of unstable storage, complicated preparation, poor batch-to-batch reproducibility, etc., and a large amount of co-emulsifier such as sodium deoxycholate, etc. may be added in the preparation of the nanoemulsion, and multiple administrations may cause severe gastrointestinal side reactions, so it is necessary to design a safe and simple oral dosage form of the triglyceride-like prodrug.
The invention content is as follows:
aiming at the problem of poor oral absorption of docetaxel, the invention provides a docetaxel-stearic acid triglyceride prodrug based on a natural triglyceride lymph transport mechanism, and prepares the prodrug into a lipid preparation of the docetaxel-stearic acid triglyceride prodrug, the preparation process is simple, the repeatability is strong, the industrialization is convenient, the prodrug is uniform and stable, the prodrug lipid preparation can promote the lymph transport of insoluble drugs and improve the oral bioavailability of the drugs, and on the basis, a reduction sensitive connecting bond is introduced, so that the antitumor drugs can be specifically released at a target site while the oral absorption of the antitumor drugs is promoted, the curative effect is improved, and the toxicity is reduced.
A first object of the present invention is to provide docetaxel-triglyceride stearate prodrug or geometric isomer, pharmaceutically acceptable salt, hydrate, solvate thereof:
Figure BDA0003045543880000021
the second object of the present invention is to provide a process for the preparation of the above compounds. The preparation method comprises the following steps:
(a) Synthesis of 1, 3-glyceryl distearate: dissolving a certain amount of stearic acid in dichloromethane, reacting with 1, 3-dihydroxyacetone under the catalysis of 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride (EDCI) and 4-Dimethylaminopyridine (DMAP) to generate ester, and performing hydrogenation reaction by sodium borohydride to obtain the 1, 3-glyceryl distearate.
Figure BDA0003045543880000022
(b) Dissolving dithiodibutyrate in a proper amount of acetic anhydride, stirring at room temperature to obtain dithiodibutyrate anhydride, reacting with 1, 3-glyceryl distearate in the presence of EDCI and DMAP to obtain an intermediate product, and esterifying with docetaxel in the presence of DMAP and EDCI to obtain a target prodrug.
Figure BDA0003045543880000031
The third purpose of the invention is to provide a simple and safe pharmaceutical composition containing a docetaxel-triglyceride stearate prodrug, wherein the pharmaceutical composition is a lipid preparation of the docetaxel-triglyceride stearate prodrug, and comprises the docetaxel-triglyceride stearate prodrug and auxiliary materials, the auxiliary materials comprise a phospholipid emulsifier, a short-chain alcohol or ether co-emulsifier and a liquid oil phase, and 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 alcohol or ether co-emulsifier accounts for 5-25% of the lipid preparation in percentage by weight; the docetaxel-stearic acid triglyceride prodrug accounts for 1 to 10 percent of the total weight of the auxiliary materials.
Further, the pharmaceutical composition comprises, by weight, 57-85% of the lipid preparation in liquid oil phase, 9-33% of the lipid preparation in phospholipid emulsifier, and 5-10% of the lipid preparation in short-chain alcohol or ether co-emulsifier;
preferably, the pharmaceutical composition of the invention comprises, by weight, 57% -75% of the lipid preparation in the liquid oil phase, 20% -33% of the lipid preparation in the phospholipid emulsifier, and 5% -10% of the lipid preparation in the short-chain alcohol or ether co-emulsifier.
Furthermore, the docetaxel-stearic acid triglyceride prodrug accounts for 2-5% of the total weight of the auxiliary materials;
preferably, the docetaxel-stearic acid triglyceride prodrug accounts for 2-4% of the total weight of the auxiliary materials.
Wherein the phospholipid is yolk lecithin, soybean lecithin, synthetic phospholipids such as 1, 2-dicaprylyl-sn-glycero-3-phosphocholine, 1, 2-didecanoyl-sn-glycero-3-phosphocholine, preferably yolk lecithin;
the liquid oil is selected from long chain triglycerides, mixed long chain glycerides, medium chain triglycerides, or combinations 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 and Peceol, preferably olive oil.
The short-chain alcohol or ether coemulsifier is diethylene glycol monoethyl ether (Transcutol HP), ethanol, propylene glycol and the like, wherein Transcutol HP is preferred.
Furthermore, the weight ratio of the short-chain alcohol or ether co-emulsifier to the phospholipid emulsifier to the liquid oil phase is 1:2-4:5-20.
The lipid preparation of the docetaxel-stearic acid triglyceride prodrug is prepared by the following method:
mixing docetaxel-triglyceride stearate prodrug, short-chain alcohol or ether co-emulsifier, phospholipid emulsifier and liquid oil phase, and performing ultrasonic treatment to uniformly mix the auxiliary material and prodrug to obtain the prodrug lipid preparation.
Or dissolving phospholipid emulsifier and short chain alcohol or ether coemulsifier in liquid oil, and mixing by ultrasonic to obtain uniform blank preparation. And adding the docetaxel-stearic acid triglyceride prodrug into the blank preparation, and performing ultrasonic treatment to completely dissolve the prodrug in the blank preparation to obtain the uniform lipid oral preparation.
When the phospholipid is egg yolk lecithin, the co-emulsifier is diethylene glycol monoethyl ether and the liquid oil is olive oil, 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 the docetaxel-stearic triglyceride prodrug has the optimal particle size and particle size distribution, has better stability, and can obviously improve the oral bioavailability of the docetaxel.
According to the invention, liquid oil (especially olive oil), phospholipid and diethylene glycol monoethyl ether are uniformly mixed to prepare a blank lipid preparation for the first time, and the synthesized reduction-sensitive docetaxel-stearic acid triglyceride prodrug is dissolved in the preparation, so that the preparation method of the lipid preparation is simple, high-temperature heating, probe ultrasound and other procedures are not needed, only simple stirring or ultrasonic dissolution is needed, the process is simple, industrialization is facilitated, and the repeatability is strong; the obtained lipid preparation is in a uniform oil solution state, and is more stable and convenient to store compared with the thermodynamically unstable state of the nano preparation; the use of sodium deoxycholate is avoided, and the auxiliary materials used by the lipid preparation provided by the invention are all FDA approved pharmaceutical auxiliary materials, so that gastrointestinal toxicity cannot be caused after oral administration; the drug loading capacity is greatly improved, and the preparation and the compliance of patients can be greatly improved; in addition, the lipid preparation is highly suitable for the designed docetaxel-stearic triglyceride prodrug, and the oral bioavailability of docetaxel is obviously improved.
The invention has the advantages that:
1. the invention designs the docetaxel-stearic acid triglyceride prodrug based on the mechanism of natural triglyceride absorption, promotes lymphatic transport of docetaxel, avoids first pass effect, and further improves oral absorption of docetaxel.
2. The invention connects the triglyceride skeleton and the insoluble drug by a reduction-sensitive disulfide bond connecting bridge, promotes the oral absorption of the drug, enables the drug to be specifically released at a target site, enhances the effect and reduces the toxicity.
3. The lipid oral preparation prepared by the invention has simple preparation process, easy industrialization, uniformity and stability, avoids the use of sodium deoxycholate, simplifies the preparation process and has no heating process or probe ultrasound compared with the nano emulsion in the prior art.
Drawings
FIG. 1 is a representation of docetaxel-triglyceride stearate prodrug1H-NMR spectrum.
Figure 2 is a high resolution mass spectrum of docetaxel-triglyceride stearate prodrug.
Figure 3 is the shelf stability of docetaxel-triglyceride stearate prodrug nanoemulsion of example 2.
Figure 4 is a graph of the solubility of docetaxel-triglyceride stearate prodrug in different excipients.
Figure 5 is a medium chain triglyceride based self-microemulsion screening pseudo-ternary phase diagram of docetaxel-triglyceride stearate prodrug.
Figure 6 is a medium chain based self-microemulsion and olive oil solution time profile of docetaxel-triglyceride stearate prodrug in SD rats.
Figure 7 is a time course of dosing in pharmacokinetic experiments with docetaxel-stearate triglyceride prodrug lipid formulation SD rats.
Figure 8 is a graph of the gastrointestinal toxicity of docetaxel-triglyceride stearate prodrug lipid formulations.
Detailed Description
The invention is further illustrated by the following examples, but is not limited thereto.
EXAMPLE 1 docetaxel-stearic acid triglyceride prodrug preparation (SATG)
The structure is as follows
Figure BDA0003045543880000051
Dissolving 11.12g (40 mmol) of stearic acid in dichloromethane, reacting with 1.75g (20 mmol) of 1, 3-dihydroxyacetone under the catalysis of EDCI and DMAP to generate ester, concentrating the reaction solution, washing with distilled water for 2 times, extracting an aqueous layer with chloroform, washing an organic layer with saturated common salt for 1 time, drying the organic layer with anhydrous sodium sulfate, and removing the solvent by rotary evaporation. Column chromatography separation was performed under the conditions of n-hexane-ethyl acetate (30. 2.9g (5 mmol) of the above product was dissolved in 100ml of a mixed solvent (THF: benzene: water 10. 2.1g (8.8 mmol) of 4,4' -dithiodibutanoic acid are dissolved in 9mL of acetic anhydride, reacted at room temperature for 2h, the solvent is concentrated by rotary evaporation and redissolved in an appropriate amount of anhydrous dichloromethane, and 620mg (1 mmol) of 1,3-DG,100mg of DMAP are added. Vacuum and nitrogen protection are carried out to react for 12 hours at room temperature. The solvent was removed by rotary evaporation, and column chromatography was performed to obtain an intermediate (provided that n-hexane: ethyl acetate = 15. 680mg (0.83 mmol) of the intermediate product are dissolved in an appropriate amount of anhydrous dichloromethane, 175mg (0.9 mmol) of EDCI,45mg (0.36 mmol) of DMAP and 803mg (1 mmol) of docetaxel are added and the reaction is carried out for 48 hours at room temperature under vacuum and nitrogen protection. And (4) removing the solvent by rotary evaporation, and separating a preparation liquid phase to obtain a final product (the preparation liquid phase condition is pure acetonitrile).
Measurement by nuclear magnetic resonance1H-NMR Hydrogen Spectroscopy to determine the structure of the prodrug of example 1, the solvent chosen was CDCl3The results are shown in the figure1. Analyzing corresponding characteristic peaks: docetaxel characteristic peak: 1H NMR (400mhz, cdcl3) δ 8.12 (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-glyceryl distearate skeleton characteristic peaks: 1.27 (d, J =9.9hz, 56h), 0.88 (t, J =6.7hz,6 h);
the molecular weight of the prodrug of example 1 was determined by high resolution mass spectrometry and the results are shown in figure 2: ESI-HRMS Calcd. For C90H139NO21S2Cl[M+Cl-]1668.9022found 1696.8909. The specific synthetic route of the prodrug is shown as follows:
Figure BDA0003045543880000061
Figure BDA0003045543880000071
example 2 stability of SATG nanoemulsion
12mg of docetaxel-triglyceride stearate prodrug was dissolved in 400mg of olive oil and preheated to 60 ℃. 120mg of egg yolk lecithin and 40mg of sodium deoxycholate were weighed, dissolved in 4ml of deionized water, and preheated to 60 ℃. Slowly dripping the oil phase into the stirred water phase, continuously stirring for 3min to form colostrum, carrying out ultrasonic treatment on the colostrum in an ice bath probe for 10min with the ultrasonic power of 500W to prepare the docetaxel-stearic acid triglyceride nanoemulsion, measuring the particle size of the emulsion in 1,2,5, 15 and 30 days, and finding that the particle size of the emulsion is obviously increased along with time and finally the phenomenon of layering occurs, thus exposing the defect that the emulsion has instability, and the result is shown in figure 3. The drug loading of the SATG nanoemulsion is only 0.258%, and therefore, the nano-emulsion prepared from the docetaxel-stearic acid triglyceride has low drug loading and poor stability.
Example 3 prescription optimization of docetaxel-triglyceride prodrug lipid formulations
Oral absorption of triglyceride-like prodrugs is highly dependent on lipid digestion. Therefore, the best carrier for SATG is the lipid-based drug delivery system (LBDDS). Among the various types of LBDDS, self-microemulsion drug delivery systems (SMEDDS) have unique advantages, including high drug loading and ease of scale-up of production.
3.1 preliminary screening and preparation of SATG self-microemulsions based on Medium Chain Triglycerides (MCT) and Long Chain Triglycerides (LCT)
Firstly, the solubility of SATG in different oil-soluble excipients was determined, and as shown in FIG. 4, SATG was found to have the highest solubility in tricaprylin (MCT), glycerol monolinoleate (Maisine 35-1), and diethylene glycol monoethyl ether (Tracnutol HP), and was used as an oil phase, an emulsifier, and a co-emulsifier. A pseudo-ternary phase diagram was created to find the optimum three ratios, as shown in figure 5, and the formulation prepared at the ratios at the spot areas, can form a self-microemulsion by simulating intragastric agitation. After further optimization, it was found that when MCT/Maisine 35-1/Transcutol HP =68.2/16.1/15.7 (by weight), the emulsion formed was smaller and more uniform in particle size.
Following PDI screening based on solubility and particle size of the emulsion formed, a medium chain triglyceride based self-microemulsion drug delivery system was developed. According to the solubility of the prodrug in various auxiliary materials, the total weight ratio of the docetaxel-triglyceride stearate prodrug to the auxiliary materials is 1:25 preparing docetaxel-stearin triglyceride lipid preparation.
In vivo pharmacokinetics experiments were performed according to the above formulation, while the olive oil solution (long chain triglyceride, LCT) group of SATG was set for comparison. As shown in FIG. 6, the AUC (0-8 h) of the SATG olive oil (LCT) solution group was significantly increased compared to SMEDDS based on MCT (medium chain triglyceride), and it was revealed that the formulation designed according to the pseudo-ternary phase diagram did not exhibit good pharmacokinetic properties in vivo although the emulsion particle size formed in the pharmaceutical formulation was small and uniform. While the LCT based formulation showed better pharmacokinetic properties in vivo than the MCT based formulation, so the long chain triglyceride LCT is more suitable for the design of self-microemulsions of SATG.
The results show that: the primary mechanism of SATG is to mimic the process of oral absorption of long chain triglycerides by digestion, absorption into intestinal cells, and re-esterification into chylomicrons, transport to the lymphatic system and then circulation. Co-administration with LCT, rather than MCT, enhances the solubility of SAMG (the product obtained after digestion of SATG by intestinal fluids) in the intestine, thereby improving oral absorption efficiency.
Therefore, the present invention selects long chain triglyceride LCT as the oil phase for further experiments.
3.2 prescription screening of Long chain triglyceride-based SATG lipid formulations
3.2.1 selection of oil phase
Based on the above results, the solubility of SATG in several LCT-based vegetable oils was determined in order to ensure the drug loading of the lipid formulations, and the results are shown in Table 1.
TABLE 1 solubility of SATG in several vegetable oils
Figure BDA0003045543880000081
The oil phase of the SATG lipid preparation is selected from olive oil, based on the solubility results.
3.2.2 selection of Co-emulsifiers
Phospholipids may help improve the oral absorption of the triglyceride-like prodrugs, so egg yolk lecithin was initially selected as the emulsifier and co-emulsifiers were screened. Dissolving the emulsifier and the co-emulsifier in an oil phase, wherein the oil phase comprises: emulsifier: the mass ratio of the auxiliary emulsifier is 65:25: and 10, uniformly mixing by using ultrasonic waves to obtain a uniform blank preparation.
200mg of SATG prodrug was added to 1g of the above blank formulation, and the equilibrium solubility of the SATG prodrug in the blank formulation was determined.
Taking 30mg of SATG prodrug, adding 1g of the blank preparation, carrying out ultrasonic treatment to completely dissolve the prodrug in the blank preparation to obtain a uniform lipid oral preparation, adding 500mg of each drug-containing preparation into 5mL of simulated gastric juice, stirring for 5min to simulate gastric peristalsis, measuring the particle size and distribution, and showing the results of different auxiliary emulsifiers in table 2.
TABLE 2 SATG lipid formulation Properties prepared with different coemulsifiers
Figure BDA0003045543880000091
The results show that different coemulsifiers are added to prepare lipid preparations with different properties, and when the coemulsifier is Transcutol HP, the solubility of the prodrug is highest, so that the optimal drug loading can be obtained, and the prepared SATG prodrug lipid preparation has the optimal drug loading, particle size and distribution.
3.2.3 selection of Phospholipids
The method comprises the steps of taking olive oil as an oil phase, taking Transcutol HP as an auxiliary emulsifier, screening phospholipid types, selecting egg yolk lecithin, soybean lecithin, 1, 2-dioctanoyl-sn-glycerol-3-phosphorylcholine and 1, 2-didecanoyl-sn-glycerol-3-phosphorylcholine, preparing an SATG lipid preparation according to a method of 3.2.2, measuring the equilibrium solubility of the SATG prodrug in a blank preparation, adding 500mg of each drug-containing preparation into 5mL of simulated gastric juice, stirring for 5min to simulate gastric peristalsis, measuring the particle size and distribution, wherein indexes of lipid preparations prepared from different phospholipids have no significant difference, and the egg yolk lecithin is relatively good.
EXAMPLE 4 preparation of SATG prodrug lipid formulations
The SATG prodrug lipid formulation was prepared according to the method 3.2.2 in example 3 to give prodrug lipid formulations of different formulations, as shown in Table 3. Adding 500mg lipid preparation into 5mL simulated gastric juice, stirring for 5min to simulate gastric motility, measuring particle size and distribution, mixing the lipid preparation with the simulated gastric juice mixture 1mL and 5mL pancreatic juice-bile, incubating for 2h, centrifuging at low temperature (10000rpm, 10min), taking the middle layer liquid, measuring the digested particle size and the SATG content ratio in the middle layer, and obtaining the results shown in Table 4:
TABLE 3 SATG lipid formulations of different formulations
Figure BDA0003045543880000092
Figure BDA0003045543880000101
TABLE 4 Properties of SATG lipid formulations of different formulations
Figure BDA0003045543880000102
As shown in table 4, when the ratio of olive oil is less than 50% (formula 1-3), the ratio of egg yolk lecithin and Transcutol HP is high, and the solubility to SATG is high, and therefore the drug loading is high, and further, due to the strong surface activity of the emulsifier and co-emulsifier, the particle size is small and uniform after mixing and stirring with simulated gastric juice, which is similar to the in vitro results of the self-microemulsion preparation based on MCT, but when mixing with pancreatic juice-bile to simulate digestion, the particle size of the aqueous phase of the intermediate layer is large, and the ratio of the SATG prodrug in the intermediate layer is less than 50%, for the triglyceride-like lipid preparation, the initial in vitro particle size does not accurately predict in vivo oral absorption, while the intermediate aqueous phase after simulated digestion corresponds to the dissolved phase of intestinal juice in vivo, the smaller particle size is, the larger ratio of the prodrug in the intermediate aqueous phase is likely to facilitate in vivo absorption. When the percentage of olive oil is more than 80% and the percentage of egg yolk lecithin is less than 20%, although the result of in vivo simulation is still in an acceptable range, the initial simulated gastric juice emulsified particle size is more than 300nm, which may delay the subsequent intestinal digestion process and further influence the oral absorption; when trancutol hp is less than 5%, olive oil is incompatible with egg yolk lecithin, and a uniform lipid preparation cannot be formed.
When the content of olive oil is 30-85%, the content of egg yolk lecithin is 9-45% and the content of trancolHP is 5-25%, a uniform and stable lipid preparation can be formed, and the particle size of the lipid preparation is less than 400nm.
When the olive oil accounts for 57-85%, the egg yolk lecithin accounts for 9-33% and the trancolHP accounts for 5-10%, the particle sizes of the prepared lipid preparations are smaller than 400nm, the SATG accounts for a large proportion in the middle layer, and the lipid preparations are beneficial to absorption in vivo.
When the olive oil accounts for 57-75%, the egg yolk lecithin accounts for 20-33%, and the trancolHP accounts for 5-10%, the particle size of the prepared lipid preparation is less than 300nm, and the lipid preparation is most beneficial to absorption in vivo.
The lipid preparation of each formula has no layering and precipitation in 15 days, and has good stability.
EXAMPLE 5 preparation of SATG prodrug lipid formulations
300mg of egg yolk lecithin is weighed and dissolved in 1g of olive oil, 100mg of diethylene glycol monoethyl ether is added at the same time, and the mixture is evenly mixed by ultrasonic, so that a uniform blank preparation is obtained. 40mg of the prodrug described in example 1 was weighed out accurately, 1g of the blank preparation was added, and the prodrug was dissolved completely therein by sonication to obtain a homogeneous lipid preparation of SATG prodrug. Precisely weighing a certain amount of docetaxel parent drug, and adding into the blank preparation to obtain the docetaxel lipid preparation.
EXAMPLE 6 pharmacokinetic experiments on lipid formulations of docetaxel-triglyceride SATG prodrugs
SD rats were used as a model, the lipid preparation containing docetaxel-triglyceride prodrug (SATG LP) and the lipid preparation containing docetaxel (DTX LP) of example 5 were orally administered, the relative dose of docetaxel was 10mg/kg, orbital blood sampling was timed, the concentration of the mother drug of docetaxel in plasma was measured, and the plotting of the drug-time curve and the calculation of the corresponding pharmacokinetic parameters were performed based on the measured concentrations, respectively (table 5). To calculate the absolute bioavailability, docetaxel solutions were administered intravenously at a dose of 5mg/kg and the content of docetaxel in the plasma was determined. As shown in FIG. 7 and Table 5, the area under the drug-time curve (AUC) for the prodrug lipid formulation group relative to docetaxel lipid formulation0-24) Compared with the parent drug lipid preparation, the preparation method is remarkably improved. The oral bioavailability of the prodrug is calculated by the data of intravenous docetaxel, and the absolute bioavailability of the prodrug reaches 17.8%. In addition, the total amount of docetaxel in blood of an SATG prodrug lipid preparation group is determined by a fragmentation mode, the absolute bioavailability of the total amount of docetaxel in a prodrug group is calculated to reach 121%, and the oral absorption of docetaxel is obviously improved.
TABLE 5 main pharmacokinetic parameters of docetaxel triglyceride prodrugs
Figure BDA0003045543880000121
EXAMPLE 7 gastrointestinal toxicity of docetaxel-triglyceride SATG prodrug lipid formulations
The cytotoxicity of the SATG prodrug lipid preparation on human clonal colon adenocarcinoma cells (CaCo-2) was examined by the MTT method. Cells were seeded into 96-well plates at a density of 1000 cells/well and placed in an incubator for 24h to adhere. Paclitaxel solutions of various concentrations, the SATG nanoemulsion prepared in example 2, and the SATG prodrug lipid formulation prepared in example 5 were added after the cells were attached to the wall. After the medicine is added for 12 hours, the cell survival rate is measured by an MTT method.
The results are shown in fig. 8, the cytotoxicity of the SATG prodrug lipid preparation to human colon adenocarcinoma cells is weaker than that of the taxol solution and the nanoemulsion, which indicates that the prodrug lipid preparation has no obvious damage to gastrointestinal tract cells, and the used auxiliary materials and the proportioning have no gastrointestinal toxicity.

Claims (10)

1. Docetaxel-stearate triglyceride prodrug or geometric isomer, pharmaceutically acceptable salt, hydrate, solvate thereof:
Figure FDA0003045543870000011
2. the method of preparing a docetaxel-triglyceride stearate prodrug, or a pharmaceutically acceptable salt, of claim 1, comprising the steps of:
(a) Synthesis of 1, 3-Glycerol distearate: dissolving stearic acid in dichloromethane, reacting with 1, 3-dihydroxyacetone under the catalysis of 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride and 4-dimethylaminopyridine to generate ester, and performing hydrogenation reaction by sodium borohydride to obtain 1, 3-glyceryl distearate;
(b) Dissolving dithiodibutanoic acid in acetic anhydride, stirring at room temperature to obtain dithiodibutanoic anhydride, reacting with 1, 3-distearate glyceride in the presence of 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride and 4-dimethylaminopyridine to obtain an intermediate product, and then carrying out esterification reaction with docetaxel to obtain the target prodrug.
3. The lipid preparation of the docetaxel-triglyceride stearate prodrug is characterized by comprising the docetaxel-triglyceride stearate prodrug or a geometric isomer, a pharmaceutically acceptable salt, a hydrate, a solvate and auxiliary materials of claim 1, wherein the auxiliary materials comprise phospholipid emulsifier, short-chain alcohol or ether co-emulsifier and liquid oil phase, and 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 alcohol or ether co-emulsifier accounts for 5% -25% of the lipid preparation according to weight percentage; preferably, 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 co-emulsifier accounts for 5-10% of the lipid preparation; more preferably, the liquid oil phase comprises 57% -75% of the lipid preparation, the phospholipid emulsifier comprises 20% -33% of the lipid preparation, and the short-chain alcohol or ether co-emulsifier comprises 5% -10% of the lipid preparation.
4. A lipid formulation of docetaxel-triglyceride stearate prodrug as claimed in claim 3, wherein the docetaxel-triglyceride stearate prodrug is present in an amount of 1-10%, preferably 2-5%, more preferably 2-4% by weight of the total adjuvant.
5. The lipid formulation of docetaxel-triglyceride stearate prodrug as set forth in claim 3 or 4, wherein the phospholipid is egg yolk lecithin, soybean lecithin, synthetic phospholipid; the liquid oil is long chain triglyceride, mixed long chain glyceride, medium chain triglyceride or their combination, preferably one or more of olive oil, almond oil, canola oil, castor oil, coconut oil, corn oil, cotton seed 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 cotton seed oil, hydrogenated palm oil, hydrogenated soybean oil, partially hydrogenated soybean oil or hydrogenated vegetable oil, maisine, peceol; the short-chain alcohol or ether co-emulsifier is diethylene glycol monoethyl ether, ethanol and propylene glycol, and the weight ratio of the short-chain alcohol or ether co-emulsifier to the phospholipid emulsifier to the liquid oil phase is 1:2-4:5-20.
6. The lipid formulation of docetaxel-triglyceride stearate prodrug as claimed in any one of claims 3 to 5, wherein the lipid formulation comprises egg yolk lecithin, docetaxel-triglyceride stearate prodrug, olive oil, diethylene glycol monoethyl ether, wherein the olive oil accounts for 57% to 75% of the lipid formulation, the egg yolk lecithin accounts for 20% to 33% of the lipid formulation, the diethylene glycol monoethyl ether accounts for 5% to 10% of the lipid formulation, and the docetaxel-triglyceride stearate prodrug accounts for 2% to 4% of the total amount of the adjuvant.
7. The method of preparing docetaxel-triglyceride stearate prodrug lipid formulation as set forth in any one of claims 3 to 6,
mixing docetaxel-triglyceride stearate prodrug, short-chain alcohol or ether coemulsifier, phospholipid emulsifier and liquid oil phase, and ultrasonically mixing the auxiliary material and the prodrug uniformly to obtain the docetaxel-triglyceride stearate prodrug;
or dissolving phospholipid emulsifier and short chain alcohol or ether co-emulsifier in liquid oil phase, mixing with ultrasonic wave to obtain uniform blank preparation, adding docetaxel-triglyceride stearate prodrug into the blank preparation, and dissolving the prodrug in the blank preparation by ultrasonic wave.
8. Use of the docetaxel-triglyceride stearate prodrug of claim 1 or a geometric isomer, a pharmaceutically acceptable salt, a hydrate, a solvate or lipid formulation of the docetaxel-triglyceride stearate prodrug of any one of claims 3 to 6 for the preparation of an anti-tumor medicament.
9. Use of the docetaxel-triglyceride stearate prodrug of claim 1 or a geometric isomer, a pharmaceutically acceptable salt, a hydrate, a solvate or docetaxel-triglyceride stearate prodrug lipid formulation of any one of claims 3 to 6 for the preparation of an oral drug delivery system.
10. Use of the docetaxel-triglyceride stearate prodrug of claim 1 or a geometric isomer, a pharmaceutically acceptable salt, hydrate, solvate or lipid formulation of docetaxel-triglyceride stearate prodrug of any one of claims 3-6 for the manufacture of a medicament for improving the therapeutic effect and reducing toxicity.
CN202110471401.5A 2021-04-29 2021-04-29 Docetaxel-stearic acid triglyceride prodrug and preparation of lipid preparation thereof Pending CN115252802A (en)

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