CN115252801A - Preparation of docetaxel-oleic acid triglyceride prodrug and lipid preparation - Google Patents
Preparation of docetaxel-oleic acid triglyceride prodrug and lipid preparation Download PDFInfo
- Publication number
- CN115252801A CN115252801A CN202110471385.XA CN202110471385A CN115252801A CN 115252801 A CN115252801 A CN 115252801A CN 202110471385 A CN202110471385 A CN 202110471385A CN 115252801 A CN115252801 A CN 115252801A
- Authority
- CN
- China
- Prior art keywords
- docetaxel
- oil
- preparation
- prodrug
- lipid
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Images
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—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
- 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
- A61K47/543—Lipids, e.g. triglycerides; Polyamines, e.g. spermine or spermidine
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- 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
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—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
- A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
- A61K47/08—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing oxygen, e.g. ethers, acetals, ketones, quinones, aldehydes, peroxides
- A61K47/10—Alcohols; Phenols; Salts thereof, e.g. glycerol; Polyethylene glycols [PEG]; Poloxamers; PEG/POE alkyl ethers
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—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
- A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
- 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
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—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
- A61K47/44—Oils, fats or waxes according to two or more groups of A61K47/02-A61K47/42; Natural or modified natural oils, fats or waxes, e.g. castor oil, polyethoxylated castor oil, montan wax, lignite, shellac, rosin, beeswax or lanolin
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/0012—Galenical forms characterised by the site of application
- A61K9/0053—Mouth and digestive tract, i.e. intraoral and peroral administration
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/10—Dispersions; Emulsions
- A61K9/127—Liposomes
- A61K9/1277—Processes for preparing; Proliposomes
- A61K9/1278—Post-loading, e.g. by ion or pH gradient
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D305/00—Heterocyclic compounds containing four-membered rings having one oxygen atom as the only ring hetero atoms
- 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
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Veterinary Medicine (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Epidemiology (AREA)
- Organic Chemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Oil, Petroleum & Natural Gas (AREA)
- Engineering & Computer Science (AREA)
- Biophysics (AREA)
- Molecular Biology (AREA)
- Physiology (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Nutrition Science (AREA)
- Dispersion Chemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Medicinal Preparation (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention belongs to the technical field of medicines, relates to a docetaxel-oleic triglyceride prodrug and a lipid preparation thereof, and particularly relates to a docetaxel-oleic triglyceride prodrug transported by lymph mediation, a lipid preparation thereof and application thereof in preparing oral chemotherapy medicamentsThe application is as follows. The invention provides a lipid preparation of a docetaxel-oleic triglyceride prodrug, which comprises the docetaxel-oleic triglyceride 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 10-45% 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-oleic acid triglyceride prodrug accounts for 1-10% of the total weight of the auxiliary materials. The docetaxel-oleic acid triglyceride prodrug disclosed by the invention promotes lymphatic transport of docetaxel, avoids first-pass effect and further improves oral absorption of docetaxel. The preparation is uniform and stable, the preparation process is simple, and the industrialization is easy.
Description
The technical field is as follows:
the invention belongs to the technical field of medicines, relates to a docetaxel-oleic triglyceride prodrug and a lipid preparation thereof, and particularly relates to a docetaxel-oleic triglyceride prodrug transported by lymph mediation, a lipid preparation thereof and application of the docetaxel-oleic triglyceride prodrug and the lipid preparation 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 the docetaxel has low solubility, the P-glycoprotein is discharged outside, and the serious first-pass effect causes the oral bioavailability to be extremely low, and no docetaxel oral preparation on the market exists 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 enhance 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. There have been many examples of triglyceride-like prodrugs, however, the structures taught by these examples are almost ineffective in enhancing the oral delivery of the drug, one important reason being the absence of cleavable linking chains, 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, medical workers prepare docetaxel into a triglyceride prodrug and prepare the triglyceride prodrug into a nanoemulsion, and improve the oral absorption of docetaxel by simulating the characteristic of oral absorption of natural triglyceride lymph transport. However, the nanoemulsion has the disadvantages of unstable storage, tedious preparation, poor batch-to-batch reproducibility, etc., and a large amount of co-emulsifier such as sodium deoxycholate may be added in the preparation of the nanoemulsion, and multiple administrations may bring about serious gastrointestinal side reactions, so that 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-oleic acid triglyceride prodrug based on a natural triglyceride lymph transport mechanism, and prepares the prodrug into a lipid preparation of the docetaxel-oleic acid triglyceride prodrug, the preparation process is simple, the repeatability is strong, the industrialization is convenient, the properties are uniform and stable, the prodrug lipid preparation can promote the lymph transport of insoluble drugs and improve the oral availability 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-oleic acid triglyceride prodrug, or geometric isomer, pharmaceutically acceptable salt, hydrate, solvate thereof:
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-dioleoyl glyceride: oleic acid is dissolved in dichloromethane, reacts with 1, 3-dihydroxyacetone under the catalysis of 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride (EDCI) and 4-Dimethylaminopyridine (DMAP) to generate ester, and then is hydrogenated by sodium borohydride to obtain 1, 3-glyceryl dioleate.
(b) Dissolving dithiodibutyrate in a proper amount of acetic anhydride, stirring at room temperature to obtain dithiodibutyrate anhydride, reacting with 1, 3-dioleic acid glyceride in the presence of EDCI and DMAP to obtain an intermediate product, and carrying out esterification reaction with docetaxel in the presence of DMAP and EDCI to obtain a target prodrug.
The third purpose of the invention is to provide a simple and safe pharmaceutical composition containing docetaxel-triglyceride oleate prodrug, the pharmaceutical composition is a lipid preparation of docetaxel-triglyceride oleate prodrug, and comprises docetaxel-triglyceride oleate prodrug and auxiliary materials, 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 10% -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-oleic acid triglyceride prodrug accounts for 1-10% of the total weight of the auxiliary materials.
Furthermore, the pharmaceutical composition comprises, by weight, 55-85% of the lipid preparation in liquid oil phase, 10-35% 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 present invention comprises, by weight, 55% -80% of the lipid preparation in the liquid oil phase, 15% -35% 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-oleic acid triglyceride prodrug accounts for 2-4% of the total weight of the auxiliary materials.
Preferably, the docetaxel-oleic acid triglyceride prodrug accounts for 3-4% 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: 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, glycerol trioctanoate, maisine, and Peceol, wherein olive oil is preferred.
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 as follows: 1:2-4:5-20.
The lipid preparation of the docetaxel-oleic acid triglyceride prodrug is prepared by the following method:
mixing docetaxel-oleic triglyceride 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 lipid preparation of the prodrug.
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-oleic 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 emulsifier is egg yolk lecithin, the short-chain alcohol or ether co-emulsifier is diethylene glycol monoethyl ether, and the liquid oil phase is olive oil, and the liquid 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, the lipid preparation of the docetaxel-triglyceride oleate prodrug has the optimal particle size and particle size distribution, has the optimal stability, and can obviously improve the oral bioavailability of the docetaxel.
According to the invention, the liquid (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-oleic acid triglyceride prodrug is dissolved in the preparation, so that the preparation method of the lipid preparation is simple, the procedures of high-temperature heating, probe ultrasound and the like are not needed, only simple stirring or ultrasonic dissolution is needed, the process is simple, the 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 a 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-oleic acid triglyceride prodrug, and the oral bioavailability of docetaxel is obviously improved.
The invention has the advantages that:
1. the invention designs the docetaxel-oleic acid triglyceride prodrug based on a natural triglyceride absorption mechanism, promotes lymphatic transport of docetaxel, avoids first pass effect, and further improves oral absorption of docetaxel.
2. The invention uses reduction-sensitive disulfide bond connecting bridges to connect the triglyceride skeleton and the insoluble drug, promotes the oral absorption of the drug, and simultaneously enables the drug to be specifically released at a target site, thereby enhancing the efficacy and reducing 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 and simplifies the preparation process without heating process and probe ultrasound compared with the nano emulsion in the prior art.
Drawings
FIG. 1 is a docetaxel-oleate triglyceride prodrug1H-NMR spectrum.
Figure 2 is a high resolution mass spectrum of docetaxel-oleic acid triglyceride prodrug.
Figure 3 is a graph of the particle size stability of docetaxel-oleate triglyceride prodrug nanoemulsion prepared in example 2.
Figure 4 is a graph of the solubility of docetaxel-oleate triglyceride prodrug in different excipients.
Figure 5 is a medium chain triglyceride-based self-microemulsion screening pseudo-ternary phase diagram of docetaxel-oleic acid triglyceride prodrug.
Figure 6 is a medium chain based self-microemulsion and olive oil solution time profile of docetaxel-oleic acid triglyceride prodrug in SD rats.
Figure 7 docetaxel-oleic acid triglyceride prodrug lipid formulation profile in SD rats with sodium deoxycholate.
Figure 8 is a plot of docetaxel-oleate triglyceride prodrug lipid formulation with/without sodium deoxycholate (bile salt) in SD rats.
Figure 9 is a plot of the dosing time of docetaxel-oleic acid triglyceride prodrug lipid formulation in beagle pharmacokinetic experiments.
Figure 10 is a graph of the gastrointestinal toxicity of docetaxel-oleic acid triglyceride prodrug lipid formulations.
Figure 11 is a graph of in vivo oral antitumor experiments of docetaxel-oleic acid triglyceride prodrug lipid formulations.
A: tumor growth curve B: mouse body weight change plot C: tumor entity map D: and (4) tumor bearing rate.
Detailed Description
The invention is further illustrated by the following examples, but is not limited thereto.
EXAMPLE 1 docetaxel-triglyceride prodrug preparation (OATG)
The structure is as follows
Dissolving 11.28g (40 mmol) of oleic acid in dichloromethane, reacting with 1.75g (20 mmol) of 1, 3-dihydroxyacetone under the catalysis of EDCI and DMAP to generate ester overnight, 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 and redissolved in an appropriate amount of anhydrous dichloromethane by rotary evaporation, and 620mg (1 mmol) of glycerol 1, 3-dioleate, 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 Spectroscopy to determine the structure of the prodrug of example 1, the solvent chosen was CDCl3The results are shown in FIG. 1. Analyzing corresponding characteristic peaks: docetaxel characteristic peak: δ 8.12 (d, J =7.3hz, 2h), 7.61 (t, J =7.4hz, 1h), 7.51 (t, J =7.8hz, 2h), 7.36 (m, 5H), 6.25 (s, 1H); 1, 3-diglycerol acid glyceride skeleton characteristic peak: 2.06-1.98 (m, 8H), 1.28 (dd, J =19.6,7.7hz, 40h), 0.88 (t, J =7.0hz, 6H);
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 C90H135NNaO21S2[M+Na+]1652.8896 found 1652.8860。
the specific synthetic route of the prodrug is shown as follows:
EXAMPLE 2 stability of OATG nanoemulsion
12mg docetaxel-oleic acid triglyceride prodrug was dissolved in 400mg 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 primary emulsion, and performing ultrasonic treatment on the primary emulsion by using an ice bath probe for 10min at the ultrasonic power of 500W to obtain the OATG nanoemulsion. The particle size of the emulsion is measured on days 1,2,5, 15 and 30, the particle size of the emulsion is obviously increased along with the time, and finally, even the phenomenon of layering occurs, so that the defect that the emulsion is unstable is exposed, and the result is shown in figure 3, the drug loading of the OATG nanoemulsion is only 0.262 percent, and the docetaxel-oleic acid triglyceride is prepared into the nanoemulsion, so that the drug loading is low and the stability is poor.
Example 3 prescription optimization of docetaxel-triglyceride prodrug lipid formulations
Oral absorption of triglyceride-like prodrugs is highly dependent on lipid digestion. Thus, the best carrier for OATG 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 formula based on OATG self-microemulsions of Medium Chain Triglycerides (MCT) and Long Chain Triglycerides (LCT)
Firstly, the solubility of OATG in different oil-soluble auxiliary materials is measured, as shown in figure 4, the solubility of OATG in tricaprylin and diethylene glycol monoethyl ether is found to be the highest, and the OATG is used as an oil phase and a co-emulsifier, and a large number of documents report that Tween 80 can promote lymphatic transport of drugs, so Tween-80 is used as an emulsifier. A pseudo ternary phase diagram is built to find the optimum three-term ratio. As shown in fig. 5, the formulation prepared at a ratio of multiple points in the figure can form a self-microemulsion by simulating intragastric agitation. After further optimization, it was found that when tricaprylin/tween 80/Transcutol HP =26.5/63/10.5 (weight ratio), the particle size of the formed emulsion was small and more uniform.
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 each adjuvant, the total weight ratio of the docetaxel-oleic acid triglyceride prodrug to the adjuvant is 1:20 preparing docetaxel-oleic acid triglyceride lipid formulation.
In vivo pharmacokinetic experiments were performed on docetaxel-oleic acid triglyceride lipid formulations prepared according to the above formulation, while the olive oil solution (long chain triglyceride LCT) group of OATG was set for comparison.
As shown in FIG. 6, the AUC (0-8 h) of the OATG olive oil (LCT) solution group was significantly increased (2.6-fold) compared to MCT (medium chain triglyceride) -based SMEDDS, and it was found that although the emulsion particle size formed in the pharmaceutical formulation designed according to the pseudo-ternary phase diagram was smaller and more uniform, the pharmacokinetic properties in vivo were lower than those of the long chain triglyceride-based solution group, i.e., the medium chain triglyceride-based self-microemulsion did not exhibit better in vivo pharmacokinetic properties. While LCT-based formulations exhibit better in vivo pharmacokinetic properties than MCT-based formulations, long chain triglyceride LCTs are more suitable for the design of self-microemulsions for OATG.
The results show that: the main mechanism of OATG 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 OAMG (the product obtained after digestion of OATG 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 OATG lipid formulations
3.2.1 selection of oil phase
Based on the above results, in order to ensure drug loading of lipid formulations, the solubility of OATG in several LCT-based vegetable oils was determined and the results are shown in Table 1.
TABLE 1 solubility of OATG in several vegetable oils
The oil phase of the OATG lipid preparation was selected from olive oil based on solubility results.
3.2.2 selection of Co-emulsifiers
Phospholipids may help to improve the oral absorption of the triglyceride-like pro-drugs, so egg yolk lecithin was initially selected as the emulsifier and screened for co-emulsifiers. Dissolving emulsifier and co-emulsifier in oil phase, oil phase: emulsifier: the mass ratio of the auxiliary emulsifier is 65:25: and 10, uniformly mixing the components by ultrasonic treatment to obtain a uniform blank preparation.
200mg of OATG prodrug was added to 1g of the above blank and the equilibrium solubility of OATG prodrug in the blank was determined.
In addition, 30mg of OATG prodrug is taken and added with 1g of the blank preparation, the prodrug is completely dissolved in the blank preparation by ultrasonic treatment to obtain a uniform lipid oral preparation, 500mg of each drug-containing preparation is added into 5mL of simulated gastric juice and stirred for 5min to simulate gastric peristalsis, the particle size and the distribution are measured, and the results of different coemulsifiers are shown in Table 2.
TABLE 2 Properties of OATG lipid formulations prepared with different coemulsifiers
The results show that the lipid preparation prepared by adding different coemulsifiers has different properties, and when the coemulsifier is Transcutol HP, the prodrug has the highest solubility, so that the optimal drug loading can be obtained, and the prepared OATG 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 OATG lipid preparation according to a method of 3.2.2, measuring the equilibrium solubility of the OATG 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 OATG prodrug lipid formulations
OATG prodrug lipid formulations were prepared according to the method of 3.2.2 in example 3, resulting in 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 and simulated gastric juice mixture 1mL and 5mL pancreatic juice-bile, incubating for 2h, centrifuging at low temperature (10000 rpm, 10min), taking middle layer liquid, measuring the particle size after digestion, and the ratio of OATG in the middle layer, the results are shown in Table 4:
TABLE 3 OATG lipid formulations of different formulations
TABLE 4 Properties of OATG lipid formulations of different formulations
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 OATG 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 simulated gastric juice, which is similar to the in vitro results of the MCT-based self-microemulsion preparation, but when mixing with pancreatic juice-bile to simulate digestion, the particle size of the water phase of the intermediate layer is large, and the ratio of OATG 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 water phase after simulated digestion corresponds to the dissolved phase of in vivo intestinal juice, the smaller particle size is, the larger ratio of prodrug in the intermediate water phase is likely to be advantageous for in vivo absorption. When the proportion of the olive oil is more than 80% and the proportion of the egg yolk lecithin is less than 15%, although the result of in vivo simulation is still in an acceptable range, the emulsified particle size of the initial simulated gastric juice 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 10-45% 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 55% -85%, the egg yolk lecithin accounts for 10% -35%, and the trancolHP accounts for 5% -10%, a uniform and stable lipid preparation can be formed, the particle size of the lipid preparation is smaller than 400nm, the OATG accounts for a large proportion in the middle layer, and the lipid preparation is beneficial to absorption in vivo.
When the olive oil accounts for 55% -80%, the egg yolk lecithin accounts for 15% -35%, and the trancolHP accounts for 5% -10%, a uniform and stable lipid preparation can be formed, the particle size of the 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 layered precipitation in 15 days, and has good stability.
EXAMPLE 5 optimization of dosing regimens for OATG prodrug lipid formulations
300mg of egg yolk lecithin is weighed and dissolved in 1g of olive oil, and 100mg of diethylene glycol monoethyl ether is added at the same time, and the materials are mixed evenly by ultrasonic waves, so that a uniform blank preparation is obtained. 40mg of the prodrug described in example 1 was weighed out accurately, 1g of the blank formulation was added, and the prodrug was completely dissolved therein by sonication to give a homogeneous lipid preparation of OATG prodrug. Precisely weighing a certain amount of docetaxel mother drug, and adding into the blank preparation to obtain docetaxel lipid preparation.
The drug loading of the lipid preparation reaches 3.85%, is improved by 14.6 times compared with the nano-emulsion in the prior art, is a uniform oily preparation all the time in the placement process, and has no layered precipitation phenomenon.
It was found that sodium deoxycholate can help improve the oral absorption of the triglyceride-like prodrug. Lipid formulations of OATG and DTX were prepared as shown in example 5 and administered with aqueous sodium deoxycholate prior to administration to SD rats. Figure 7 shows the oral plasma profile after administration of LCT-based SMEDDS with sodium deoxycholate solution as a supplement to OATG and DTX, corresponding pharmacokinetic parameters are given in table 5. AUC (0-24 hours) for the group of OATG prodrugs given concurrently with the sodium deoxycholate solution showed a 1.86-fold increase compared to DTX. In addition, C max of OATG group (221.237 + -82.95 ng mL-1) was 6.9 times higher than that of DTX group (31.984 + -13.852 ng mL-1).
TABLE 5 oral pharmacokinetic parameters of LCT-based SMEDDS formulations
However, the use of sodium deoxycholate may lead to impaired gastrointestinal function. Therefore, we investigated the necessity of sodium deoxycholate in the formulation. The oral plasma profiles of SD rats in OATG and DTX groups after removal of sodium deoxycholate from the above formulations are shown in FIG. 8. AUC (0-24 hr) was not significantly different between OATG groups using (633.916 + -164.122 μ g Lh-1) and no (697.1 + -71.3 μ g L-1) sodium deoxycholate. The OATG group had a relative oral bioavailability (Frel) of 247.5% and an absolute oral bioavailability (Fab) of 37%, in sharp contrast to 15% in the DTX group.
Example 6 beagle pharmacokinetic experiments on docetaxel-oleic acid triglyceride prodrug lipid formulations
A beagle dog is taken as a model, a hard capsule containing a docetaxel-oleic acid triglyceride prodrug lipid preparation (OATG LP) and a hard capsule containing a docetaxel lipid preparation (DTX LP) are orally taken, the dose is 3mg/kg, blood is collected from the orbit at regular time, the concentration of a docetaxel mother drug in blood plasma is measured, and the drawing of a drug-time curve and the calculation of corresponding pharmacokinetic parameters are respectively carried out according to the measured concentration (table 6). To calculate the absolute bioavailability, docetaxel solutions were administered intravenously at a dose of 1mg/kg and the content of docetaxel in the plasma was determined. As shown in figure 9 and Table 6, the area under the drug-time curve (AUC) for the prodrug lipid formulation group relative to the docetaxel lipid formulation0-24) Compared with the parent drug lipid preparation, the preparation method has the advantages of obvious improvement. The oral bioavailability of the prodrug was calculated from the data of intravenous docetaxel and the absolute bioavailability of the prodrug reached 41.08%.
TABLE 6 main pharmacokinetic parameters of docetaxel triglyceride prodrugs
EXAMPLE 7 gastrointestinal toxicity of docetaxel-triglyceride OATG prodrug lipid formulations
The cytotoxicity of lipid preparations of OATG prodrugs on human cloned colon adenocarcinoma cells (CaCo-2) was investigated using 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. Serial concentrations of paclitaxel solutions, the OATG nanoemulsion prepared in example 2, and the OATG prodrug lipid formulation prepared in example 5 were added after cells adhered. After the medicine is added for 12 hours, the cell survival rate is measured by an MTT method.
The results are shown in fig. 10, the cytotoxicity of the OATG prodrug lipid preparation to human colon adenocarcinoma cells is weaker than that of a docetaxel solution and a nanoemulsion, which indicates that the prodrug lipid preparation has no obvious damage to gastrointestinal tract cells, and the used auxiliary materials and the mixture ratio have no gastrointestinal toxicity.
EXAMPLE 8 pharmacodynamic behavior of docetaxel triglyceride prodrug lipid formulations
Orally taking a docetaxel triglyceride lipid preparation, a docetaxel lipid preparation and a docetaxel solution by using a 4T1 in-situ tumor-bearing Balb/c mouse model, wherein the administration dose is 10mg/kg (docetaxel equivalent dose), and the administration is carried out every day; setting a docetaxel solution intravenous injection group as a positive control, wherein the administration dose is 10mg/kg, and the docetaxel solution is administered once every two days; PBS group was set as blank control. The results are shown in fig. 11, the docetaxel-triglyceride prodrug lipid preparation group has the smallest tumor volume, has a significant difference with the control group, and has no weight loss phenomenon, which indicates that the docetaxel-triglyceride prodrug lipid preparation group has reliable safety and good anti-tumor effect.
Claims (10)
1. Docetaxel-oleic acid triglyceride prodrug or geometric isomer, pharmaceutically acceptable salt, hydrate, solvate thereof:
2. the method of preparing a docetaxel-oleic triglyceride prodrug of claim 1, comprising the steps of:
(a) Synthesis of 1, 3-diolein: dissolving oleic 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 dioleate;
(b) Dissolving dithiodibutanoic acid in acetic anhydride, stirring at room temperature to obtain dithiodibutanoic anhydride, reacting with 1, 3-dioleic acid glyceride in the presence of 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride and 4-dimethylaminopyridine to obtain an intermediate product, and carrying out esterification reaction with docetaxel to obtain the dithiodibutanoic anhydride.
3. The lipid preparation of the docetaxel-oleic acid triglyceride prodrug is characterized by comprising the docetaxel-oleic acid triglyceride prodrug as claimed in claim 1 and auxiliary materials, wherein the auxiliary materials comprise a phospholipid emulsifier, a short-chain alcohol or ether co-emulsifier and a liquid oil phase, 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 in percentage by weight; preferably, the liquid 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; more preferably, the liquid 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.
4. Lipid formulation of docetaxel-oleate triglyceride prodrugs of claim 3, wherein docetaxel-oleate triglyceride prodrug represents 1-10%, preferably 2-4% of the total weight of the adjuvant.
5. The lipid formulation of docetaxel-oleic triglyceride prodrug of claim 3 or 4, wherein the phospholipid is egg 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, 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 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 as follows: 1:2-4:5-20.
6. The lipid formulation of docetaxel-oleate triglyceride prodrugs of any one of claims 3 to 5, wherein the lipid formulation comprises egg yolk lecithin in an amount of 55% to 80% of the lipid formulation, docetaxel-oleate triglyceride prodrug, olive oil in an amount of 15% to 35% of the lipid formulation, diethylene glycol monoethyl ether in an amount of 5% to 10% of the lipid formulation, and docetaxel-oleate triglyceride prodrug in an amount of 3% to 4% of the total amount of the adjuvant.
7. The method for preparing lipid preparation of docetaxel-oleic acid triglyceride prodrug as claimed in any one of claims 3 to 6, wherein docetaxel-oleic acid triglyceride prodrug, short-chain alcohol or ether coemulsifier, phospholipid emulsifier, liquid oil are mixed in oil phase, and the mixture of adjuvant and prodrug is homogenized by ultrasonic wave;
or dissolving phospholipid emulsifier and short chain alcohol or ether co-emulsifier in liquid oil, mixing with ultrasonic wave to obtain uniform blank preparation, adding docetaxel-oleic acid triglyceride prodrug into the blank preparation, and dissolving the prodrug in the blank preparation by ultrasonic wave.
8. Use of docetaxel-oleate triglyceride prodrug of claim 1 or a geometric isomer, a pharmaceutically acceptable salt, a hydrate, a solvate thereof or a docetaxel-oleate triglyceride prodrug lipid formulation of any one of claims 3 to 6 for the preparation of an oral drug delivery system.
9. Use of docetaxel-oleate triglyceride prodrug of claim 1 or geometric isomer, pharmaceutically acceptable salt, hydrate, solvate or docetaxel-oleate triglyceride prodrug lipid formulation of any one of claims 3 to 6 for the preparation of an antitumor drug.
10. Use of the docetaxel-oleate triglyceride prodrug of claim 1 or a geometric isomer, a pharmaceutically acceptable salt, a hydrate, a solvate thereof or a docetaxel-oleate triglyceride prodrug lipid formulation of any one of 3 to 6 for the preparation of a medicament for improving the therapeutic effect and reducing the toxicity thereof.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110471385.XA CN115252801A (en) | 2021-04-29 | 2021-04-29 | Preparation of docetaxel-oleic acid triglyceride prodrug and lipid preparation |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110471385.XA CN115252801A (en) | 2021-04-29 | 2021-04-29 | Preparation of docetaxel-oleic acid triglyceride prodrug and lipid preparation |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN115252801A true CN115252801A (en) | 2022-11-01 |
Family
ID=83745982
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202110471385.XA Pending CN115252801A (en) | 2021-04-29 | 2021-04-29 | Preparation of docetaxel-oleic acid triglyceride prodrug and lipid preparation |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN115252801A (en) |
-
2021
- 2021-04-29 CN CN202110471385.XA patent/CN115252801A/en active Pending
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| JP7229967B2 (en) | lymphotropic prodrug | |
| Nanjwade et al. | Functions of lipids for enhancement of oral bioavailability of poorly water-soluble drugs | |
| US20050118254A1 (en) | Microemulsion preconcentrate | |
| Pandey et al. | Lipids and surfactants: The inside story of lipid-based drug delivery systems | |
| CN109851593B (en) | Triglyceride prodrug based on lymph-mediated transport and preparation method thereof | |
| JP2018184458A (en) | Calcifediol soft capsules | |
| US20080146671A1 (en) | Microemulsion preconcentrate comprising a renin inhibitor | |
| JP6158513B2 (en) | Taxol / steroid compound | |
| HUE027135T2 (en) | Pharmaceutical compositions comprising a camtothecin derivate | |
| CN106999440A (en) | The pharmaceutical composition for being used to be administered orally containing taxane | |
| US20080279940A1 (en) | Microemulsions of Cannabinoid Receptor Binding Compounds | |
| CN104292289A (en) | Galactose ligand and its application in liver targeting liposome | |
| CN113980029B (en) | SN38 triglyceride prodrug, lipid preparation, and preparation method and application thereof | |
| UA111074C2 (en) | Pharmaceutical formulation containing phenylamino pyrimidine derivative as active substance | |
| WO2017049821A1 (en) | Water-soluble docetaxel anticancer drug compound and preparation method and use thereof | |
| CN115252801A (en) | Preparation of docetaxel-oleic acid triglyceride prodrug and lipid preparation | |
| KR20050084572A (en) | Derivatives comprising sterols and/or stanols and specific classes of anti-inflammatory agents and use thereof in treating or preventing cardiovascular disease | |
| Baldi et al. | Armamentarium of nanoscaled lipid drug delivery systems customized for oral administration: in silico docking patronage, absorption phenomenon, preclinical status, clinical status and future prospects | |
| CN115252802A (en) | Docetaxel-stearic acid triglyceride prodrug and preparation of lipid preparation thereof | |
| CN108310390B (en) | Docetaxel linoleate and fat emulsion and application thereof | |
| CN115252800A (en) | Prodrug of taxol triglyceride transferred by lymph mediation and lipid preparation | |
| CN107072966B (en) | Capsaicin-vitamin E prodrug self-assembly nanoparticles and preparation method and application thereof | |
| Miao et al. | Enhancing Oral Performance of Paclitaxel Lipid‐Mimic Prodrug via Modulating Type of Fatty Acids | |
| CN116514743A (en) | Paclitaxel-fatty acid prodrug, self-microemulsion preparation, and preparation methods and applications thereof | |
| CN104293859A (en) | Enzymatic synthesis method of galactose ligand molecule |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination |