A celecoxib-vitamin e prodrug, its preparation and use

Abstract

The present application relates to a celecoxib-vitamin E prodrug. The celecoxib-vitamin E prodrug is prepared by connecting celecoxib with vitamin E by means of a dithio-bis-acetic acid. The catalysts used include: dicyclohexyl carbodiimide (DCC), 4-dimethylaminopyridine (DMAP). The prodrug has some characteristics: uniform distribution in a spherical shape, the average particle size is about 700 nm, the drug loading can reach 4 mg•mL ‑1 The prodrug can improve the bioavailability of celecoxib, and the preparation method thereof is disclosed.

Description

TECHNICAL FIELD

[0001] The present application relates to the preparation of celecoxib-vitamin E prodrug and its application in drug delivery system. BACKGROUND

[0002] Celecoxib is a second-generation specific cyclooxygenase-2 (COX-2) inhibitor, which is mainly used in the treatment of osteoarthritis, rheumatoid arthritis, ankylosing spondylitis and adult acute pain in clinic. Compared with non-selective cyclooxygenase inhibitors (aspirin, acetaminophen, etc.), celecoxib has less effect on cyclooxygenase-1 (COX-1), does not affect the synthesis of prostaglandin (PGI2) which has protective effect on gastrointestinal tract and kidney, greatly reduces the probability of causing gastrointestinal complications and kidney side effects, and can significantly reduce the recurrence rate of sporadic colorectal adenoma.

[0003] Prodrugs play a role through changes in their molecular structure. Biological prodrugs are not active themselves, and the active metabolites are in the body, which avoids the inactivation of the compound by metabolic reaction, but uses the metabolic reaction in the body to generate active compounds. The combination of prodrug technology and celecoxib can improve the properties of the drug, increase the solubility and improve the bioavailability. Therefore, the preparation of celecoxib-vitamin E prodrug can increase the solubility of celecoxib and improve the bioavailability. SUMMARY

[0004] One of the purposes of the present application is to provide a preparation method of celecoxib-vitamin E prodrug.

[0005] The second purpose of the present application is to provide the application of celecoxib-vitamin E prodrug in the preparation of anti-inflammatory analgesic and anti-colorectal cancer drugs.

[0006] The technical solution of the present application is as follows:

[0007] A celecoxib-vitamin E prodrug is formed by connecting celecoxib with one molecule of vitamin E through dithioxydiacetic acid, using celecoxib as the parent drug, and using DCC and DMAP as catalysts, and the structure of the celecoxib-vitamin E prodrug is shown as follows:

[0008]

[0009] The mass ratio of the catalyst to the celecoxib-vitamin E prodrug is 5-10:2-4:2-4.

[0010] The catalyst in the above-mentioned celecoxib-vitamin E prodrug is DCC and DMAP.

[0011] A method for preparing the celecoxib-vitamin E prodrug, comprising the following steps:

[0012] Step 1, dissolve 1 g of dithiodiglycolic acid in 10-20 mL of acetic anhydride, stir at room temperature for 3-4 hours, and then rotary evaporate to dryness. Dissolve the crude product in 15-40 mL of dichloromethane, add 10-30 mg of 4-dimethylaminopyridine (DMAP) and 500 mg of vitamin E, stir at room temperature for 2 hours, and then purify the reaction solution by silica gel column. The eluent is petroleum ether-ethyl acetate-glacial acetic acid (95:5:0.1 ~ 80:20:0.1, V / V) to obtain S-VE;

[0013] Step 2, dissolve 500 mg of S-VE in dichloromethane, add 100-400 mg of DCC and 10-30 mg of DMAP, stir at room temperature for 5 min, add 250 mg of celecoxib, stir at room temperature for two hours, and then purify the reaction solution by silica gel column. The eluent is dichloromethane-methanol (100:0-90:10 V / V) to obtain a light yellow viscous semi-solid, which is the celecoxib-vitamin E prodrug.

[0014] The method for preparing the celecoxib-vitamin E prodrug described above, wherein the catalyst is DCC and DMAP.

[0015] The celecoxib-vitamin E prodrug described above, in the preparation of anti-inflammatory analgesic and anti-colorectal cancer drugs.

[0016] Beneficial effects

[0017] The celecoxib-vitamin E prodrug of the present application is uniformly distributed in a spherical shape, with an average particle size of about 700 nm, and the drug loading capacity, calculated as celecoxib, can reach 4 mg•mL -1 . The prodrug can increase the solubility of celecoxib and significantly improve the bioavailability of celecoxib. BRIEF DESCRIPTION OF DRAWINGS

[0018] Figure 1 The H-NMR chart of the celecoxib-vitamin E prodrug in Example 1 of the present application. 1 .

[0019] Figure 2 The transmission electron microscope chart of the celecoxib-vitamin E prodrug in Example 2 of the present application.

[0020] Figure 3 The HCT-116 cell proliferation inhibition rate in Example 5 of the present application. DETAILED DESCRIPTION

[0021] The following examples are helpful for those skilled in the art to better understand the present application, but do not limit the present application in any way.

[0022] Example 1. Synthesis of celecoxib-vitamin E prodrug

[0023] Take 1 g of dithioxydiacetic acid and dissolve it in 10 mL of acetic anhydride, stir at room temperature for 3-4 hours, and then rotary evaporate at 60°C until dry after the reaction is completed. Dissolve the crude product in an appropriate amount of dichloromethane, add 20 mg of DMAP, and 500 mg of vitamin E, stir at room temperature for 2 hours. Purify the reaction solution with a silica gel column, and the eluent is petroleum ether-ethyl acetate-glacial acetic acid (95:5:0.1 ~ 80:20:0.1, V / V). About 800 mg of S-VE is obtained. Dissolve 500 mg of S-VE in an appropriate amount of dichloromethane, add 200 mg of DCC and 14 mg of DMAP, stir at room temperature for 5 min, add 250 mg of celecoxib, and stir at room temperature for two hours. Purify the reaction solution with a silica gel column, and the eluent is dichloromethane-methanol (100:0 ~ 90:10 V / V). About 500 mg of light yellow sticky semi-solid is obtained, which is the celecoxib-vitamin E prodrug.

[0024] The structure of the celecoxib-vitamin E prodrug in Example 1 is determined by nuclear magnetic resonance.

[0025] Weigh 20 mg of monomer ingredients, dissolve them in 0.6 mL of deuterated chloroform, and use a nuclear magnetic resonance analyzer for detection. 1 H-NMR spectrum is shown in Figure 1 .

[0026] Example 2. Morphology observation of celecoxib-vitamin E prodrug

[0027] The morphology of the celecoxib-vitamin E prodrug in Example 1 is observed by transmission electron microscopy. Specifically, the celecoxib-vitamin E prodrug is diluted with an appropriate amount of distilled water and then dropped onto a copper mesh, which is then dried at room temperature. A drop of 2% phosphotungstic acid is added to the sample for staining, which is then dried at room temperature. The morphology characteristics are observed by transmission electron microscopy. Figure 2 The celecoxib prodrug has a spherical distribution.

[0028] Example 3. Particle size distribution of celecoxib-vitamin E prodrug

[0029] The particle size distribution and Zeta potential of the celecoxib-vitamin E prodrug in Example 1 are measured by a laser particle size analyzer. Specifically, the celecoxib-vitamin E prodrug is prepared and diluted with an appropriate amount of distilled water, and then the particle size distribution and Zeta potential are measured by a laser particle size analyzer. The average particle size is 700 ±0.5 nm, indicating that the celecoxib-vitamin E prodrug is stable.

[0030] Example 4. Pharmacokinetic study of celecoxib-vitamin E prodrug

[0031] 24 healthy male SD rats were randomly divided into 4 groups, 6 rats in each group. The tail vein was injected at a dose of 1.5 mg·kg -1 , 50 mg·kg -1 , 50 mg·kg max , and 50 mg·kg max respectively. The rats were orally administered with celecoxib, celecoxib-vitamin E prodrug in Example 1. At the specified time points, about 0.5 mL of blood was taken from the orbit, and plasma was obtained by separation, and the drug concentration of celecoxib in rat plasma was determined by HPLC method.

[0032] After intravenous administration, the pharmacokinetic parameters were calculated:

[0033]

[0034] After oral administration, the pharmacokinetic parameters were calculated:

[0035]

[0036] The results show that after oral administration of celecoxib-vitamin E prodrug, the blood drug concentration, AUC and C max are significantly improved, the T max after oral administration is significantly prolonged, indicating that celecoxib-vitamin E prodrug significantly improves the bioavailability of celecoxib and has a sustained-release effect.

[0037] Example 5. Study on celecoxib-vitamin E prodrug promoting apoptosis of colorectal cancer cells

[0038] The colon cancer cells (HCT-116) were cultured in a 37℃, 95% humidity, 5% CO2 incubator containing 10% fetal bovine serum (FBS), and the cells were monolayer adherent growth, and the liquid was changed every 2-3 days, and the cells were subcultured according to a 1:3 ratio by 0.25% (w / v) trypsin digestion.

[0039] A certain amount of blank solution was weighed in a sterilized EP tube, dissolved with complete culture medium, and sterilized by ultraviolet irradiation for 10 min. The cells were inoculated on a 96-well culture plate at a density of 5×10 4 / mL, 100 μL per well, and when the cells were growing actively, the blank solution, celecoxib raw material and celecoxib prodrug were added, and detected after 24 h, 48 h and 72 h of culture. The detection method: 200 μL of 5 mg / mL thiazolyl blue solution (MTT) was added to each well, and incubated for 4 h. The liquid was poured out, 150 μL of dimethyl sulfoxide (DMSO) was added to each well, and gently shaken for 10 min. The absorbance (OD) value at 570 nm was detected by automatic enzyme marker. The MTT experiment showed that the celecoxib prodrug had strong cytotoxicity to HCT-116 cancer cells and could inhibit the growth of cancer cells.

[0040] The HCT-116 cell proliferation inhibition rate in vitro was calculated:

[0041] .

Claims

1. A celecoxib-vitamin E prodrug, characterized in that: It is a celecoxib-vitamin E prodrug formed by linking celecoxib to one molecule of vitamin E with dithiodiacetic acid as the parent drug. The catalysts used are dicyclohexylcarbodiimide and 4-dimethylaminopyridine. The structural formula of the celecoxib-vitamin E prodrug is shown below: The mass ratio of the catalyst to the celecoxib-vitamin E prodrug is 5-10:2-4:2-4. The celecoxib-vitamin E prodrug has an average particle size of 700nm±0.5nm and is uniformly distributed in a near-spherical shape.

2. A method for preparing the celecoxib-vitamin E prodrug of claim 1, characterized in that: Includes the following steps: Step 1: Dissolve 1g of dithionyldiacetic acid in 10-20mL of acetic anhydride, stir at room temperature for 3-4 hours, and evaporate to dryness by rotary evaporation after the reaction is complete. Dissolve the crude product in 15-40mL of dichloromethane, add 10-30mg of 4-dimethylaminopyridine and 500mg of vitamin E, and stir at room temperature for 2 hours. Purify the reaction solution by silica gel column chromatography, using petroleum ether-ethyl acetate-glacial acetic acid as the eluent (v / v) at a ratio of 95:5:0.1-80:

20. 0.1, to obtain dithionyl diacetic acid-vitamin E; Step 2: Dissolve 500 mg of dithionyl diacetic acid (Vitamin E) in dichloromethane, add 100–400 mg of dicyclohexylcarbodiimide and 10–30 mg of 4-dimethylaminopyridine, stir at room temperature for 5 min, add 250 mg of celecoxib, stir at room temperature for 2 hours, purify the reaction solution with silica gel column chromatography, and eluent is dichloromethane-methanol in a volume ratio of 100:0–90:10 to obtain a light yellow viscous semi-solid, which is the celecoxib Vitamin E prodrug.

3. The use of the celecoxib-vitamin E prodrug as described in claim 1 in the preparation of an anti-colorectal cancer drug.

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