KR102684499B1 - Mebendazole-containing solid dispersion, pharmaceutical composition comprising the solid dispersion and preparation method of the solid dispersion - Google Patents

Abstract

The present invention relates to a solid dispersion containing mebendazole and sodium dodecyl sulfate (SDS), and a pharmaceutical composition containing the same, whereby poorly soluble mebendazole exists in an amorphous state. The solubility and dissolution rate of mebendazole are increased, which has the effect of significantly improving bioavailability.

Description

Mebendazole-containing solid dispersion, pharmaceutical composition comprising the solid dispersion and preparation method of the solid dispersion}

The present invention relates to a solid dispersion containing mebendazole and a method for producing the same. Additionally, the present invention relates to a pharmaceutical composition for cancer treatment containing the above solid dispersion.

Mebendazole (MBZ) is a drug used to treat a number of parasitic infections, including ascariasis, urinariasis, hookworm infection, Medina infection, echinococcosis, trichinellosis, and giardiasis. Mebendazole acts through a mechanism that interferes with parasite β-tubulin polymerization and thereby inhibits microtubule-dependent functions.

Mebendazole has the chemical structure of [Chemical Formula 1] below.

Typically, when a drug is administered orally, the gastrointestinal tract (GI tract) is affected by two factors: solubility and permeability. However, mebendazole is a Biopharmaceutical Classification System (BCS) class II drug and has very low aqueous solubility, resulting in low bioavailability. The bioavailability of mebendazole is only 17% when administered orally to humans (Dawson M, et al.), so high doses are required to achieve the blood levels required for treatment. Moreover, the absorption of mebendazole is limited by the low dissolution of mebendazole in acidic media, resulting in low bioavailability.

Through studying the relationship between the drug's in vitro properties and in vivo parameters, it was found that the increased solubility of mebendazole in acidic media resulted in an increase in the area under the curve (AUC) and maximum concentration (Cmax). It was confirmed that bioavailability was improved (Daniel-Mwambete et al.).

In addition, mebendazole is known to have anticancer activity that inhibits the proliferation of various types of cancer cells such as colon cancer, stomach cancer, adrenal cancer, breast cancer, and lung cancer. It has been reported that mebendazole has strong anticancer activity against human cancer cells in in vitro and in vivo experiments, and that mebendazole exhibits an apoptosis effect in human lung cancer cells in a dose- and time-dependent manner (Mukhopadhyay et al. .). In addition, mebendazole shows cytotoxicity with an IC50 value of 0.39 μM against ACP-2 (cell line from gastric adenocarcinoma) and an IC50 value of 1.25 μM against ACP-03 (intestinal type) (Pinto et al. al.), the in vitro anticancer activity test results of mebendazole against melanoma were reported to have high inhibitory activity with an IC50 value of 0.30 to 0.32 (Doudican et al.).

Improvement of mebendazole water solubility plays an important role in increasing the bioavailability of mebendazole drug and improving anticancer activity effect, and increased mebendazole water solubility can reduce side effects caused by high doses of mebendazole. .

In the case of mebendazole, the solubility in water is very low, making it difficult to expect high absorption in vivo, so a method to improve the solubility and bioavailability of the drug is required.

Accordingly, as a result of research conducted by the present inventors, it was confirmed that both the solubility and dissolution parameters of mebendazole were improved by preparing mebendazole as a solid dispersion (SD), thereby completing the present invention.

As a prior art, US Patent Publication No. 2017-0209372 contains 5 to 95% (w/w) of a pharmaceutically active compound (mebendazole, niclosamide, etc.), 95 to 5% (w/w) of a stabilizer (polymer and a surfactant (sodium dodecyl sulfate, etc.)), but the composition forms a solid dispersion with mebendazole by selecting SDS (sodium dodecyl sulfate), a surfactant, as a stabilizer. is not disclosed.

Korean Patent Publication No. 10-2021-0075895 discloses a pharmaceutical composition for preventing or treating cancer containing UDCA (ursodeoxycholic acid) and benzimidazole-based compounds (mebendazole, fenbendazole, etc.), but mebendazole and There is no disclosure regarding the implementation of an amorphous solid dispersion with excellent solubility using a specific surfactant.

U.S. Patent Publication 2017-0209372 (A Method of Preparing Amorphous Solid Dispersion in Submicron Range by Co-Precipitation, published on July 27, 2017) Korean Patent Publication No. 10-2021-0075895 (Pharmaceutical composition for preventing or treating cancer, published on June 23, 2021)

Dawson M, Allan R, Watson T. The pharmacokinetics and bioavailability of mebendazole in man: a pilot study using [3H]-mebendazole. Br J Clin Pharmacol 1982;(14):453-5. Daniel-Mwambete K, Torrado S, Cuesta-Bandera C, Ponce-Gordo F, Torrado J. The effect of solubilization on the oral bioavailability of three benzimidazole carbamate drugs. Int J Pharm 2004;(272):29-36. Mukhopadhyay T, Sasaki J, Ramesh R, Roth JA. Mebendazole elicits a potent antitumor effect on human cancer cell lines both in vitro and in vivo. Clin Cancer Res 2002;(8):2963-9. Doudican N, Rodriguez A, Osman I, Orlow SJ. Mebendazole induces apoptosis via Bcl-2 inactivation in chemoresistant melanoma cells. Mol Cancer Res 2008;(6):1308-15.

The purpose of the present invention is to provide a solid dispersion containing mebendazole and a pharmaceutical composition containing the same.

Another object of the present invention is to provide a method for producing a solid dispersion containing mebendazole.

The present invention relates to a mebendazole-containing solid dispersion with improved bioavailability, comprising mebendazole and sodium dodecyl sulfate (SDS).

In the solid dispersion of the present invention, mebendazole is preferably present in an amorphous form.

In the solid dispersion, the weight ratio of mebendazole:sodium dodecyl sulfate (SDS) may be 1:5 to 15 (w/w).

In the solid dispersion, the weight ratio of mebendazole:sodium dodecyl sulfate may be 1:5-10 (w/w).

In the solid dispersion, the weight ratio of mebendazole:sodium dodecyl sulfate may be 1:7-10 (w/w).

According to another aspect of the present invention, it relates to a pharmaceutical composition comprising a mebendazole-containing solid dispersion and a pharmaceutically acceptable carrier.

According to another aspect of the present invention, there is a method for preparing a solid dispersion containing mebendazole, a) mebendazole is dissolved in formic acid to prepare a mebendazole solution, and sodium dodecyl sulfate is dissolved in distilled water to prepare sodium dodecyl. Preparing a syl sulfate solution; b) adding a mebendazole solution to the sodium dodecyl sulfate solution and stirring to prepare a mixed solution; and c) freeze-drying the mixed solution. It relates to a method for producing a solid dispersion comprising a step.

Hereinafter, the present invention will be described in more detail.

The solid dispersion of the present invention is characterized by containing mebendazole and sodium dodecyl sulfate (SDS) as active ingredients.

In the solid dispersion of the present invention, the active ingredient mebendazole is changed from a crystalline form to an amorphous state, thereby increasing the solubility and dissolution rate of mebendazole and improving bioavailability.

The solid dispersion of the present invention has a weight ratio (w/w) of mebendazole:sodium dodecyl sulfate (SDS) of 1:5-15, preferably 1:5-10, and more preferably 1:5-10. It's 7 to 10. In this way, when mebendazole: sodium dodecyl sulfate (SDS) is mixed at a specific weight ratio, the solubility and dissolution rate of mebendazole are simultaneously increased, which has the effect of significantly increasing bioavailability.

According to another aspect of the present invention, it relates to a pharmaceutical composition comprising a mebendazole-containing solid dispersion and a pharmaceutically acceptable carrier.

The pharmaceutically acceptable carrier included in the composition of the present invention includes known and used excipients, disintegrants, lubricants, etc. Examples of the excipients include lactose, mannitol, sorbitol, microcrystalline cellulose, starch, pregelatinized starch, calcium phosphate, silicon dioxide, cellulose, sodium bicarbonate, and mixtures thereof, and examples of the disintegrant include croscarmellose sodium, Crospovidone, sodium starch glycolate, low-substituted hydroxypropyl cellulose, etc. are included, and examples of the lubricant include sodium stearyl fumarate, magnesium stearate, calcium stearate, talc, etc. The pharmaceutically acceptable carrier can be appropriately selected and used by a person skilled in the art depending on the final dosage form obtained.

The dosage form of the pharmaceutical composition may be in various forms, but includes solid preparations such as granules, tablets, capsules, dry syrup, or powder, and more preferably granules, tablets, or capsules. These formulations can be prepared according to methods commonly used in the pharmaceutical field. For example, tablets can be manufactured by mixing the solid dispersion with excipients, disintegrants, lubricants, etc., or by adding excipients to the solid dispersion to form granules, mixing with excipients, disintegrants, lubricants, etc., and tableting. Capsules can also be manufactured by filling the above mixture into a capsule. Additionally, film-coating or enteric-coating may be applied to improve stability, convenience of administration, appearance, etc.

The method for producing a solid dispersion containing mebendazole of the present invention is a) dissolving mebendazole in formic acid to prepare a mebendazole solution, and dissolving sodium dodecyl sulfate (SDS) in distilled water to prepare a sodium dodecyl sulfate solution. manufacturing a; b) adding a mebendazole solution to the sodium dodecyl sulfate solution and stirring to prepare a mixed solution; and c) freeze-drying the mixed solution.

In step b), mebendazole:sodium dodecyl sulfate (SDS) has a weight ratio (w/w) of 1:5 to 15, preferably 1:5 to 10, and more preferably 1:7. It is ~10. In addition, the freeze-drying in step c) may be performed using a freeze dryer after freezing the mixed solution at about -70°C, but is not limited thereto and may be performed according to a conventional method known in the art. .

The solid dispersion of the present invention prepared by the above method increases the wettability of the drug by increasing the effective surface, and the solubilization effect of the surfactant, the absence of agglomeration of drug microcrystals, and improved dispersibility of the drug, etc., improve the dispersibility of the drug. As the dissolution rate is increased, the bioavailability of the drug is significantly increased.

In addition, the above manufacturing method has the advantage of avoiding the problems of residual solvent and toxicity when using organic solvents by not using organic solvents in the process, and providing a solid dispersion that is safe for the human body.

The solid dispersion of the present invention has the effect of significantly improving bioavailability by increasing the solubility and dissolution rate of mebendazole because poorly soluble mebendazole exists in an amorphous state. In addition, it can be confirmed that the mebendazole solid dispersion used in the present invention exhibits higher cytotoxicity than unencapsulated mebendazole in all tested cancer cells, thereby increasing the anticancer effect. Therefore, the mebendazole-containing solid dispersion can improve the anticancer effect by improving absorption and increasing cytotoxicity to cancer cells.

Figure 1 is a graph showing the solubility of mebendazole (MBZ) in a 1% aqueous solution of the carrier. Results are mean ± SD, (n=3), and all analyzes indicate statistical significance at P < 0.05 (*), P < 0.01 (**), and P < 0.001 (***).
Figure 2 is a graph showing the dissolution profile of a solid dispersion containing mebendazole (MBZ). (A) Solubility of solid dispersions (F1-F6) prepared with mebendazole and SDS surfactant in distilled water, (B) Solubility of solid dispersions (F1-F6) prepared with mebendazole and SDS surfactant in 0.1 M HCl medium (pH 1.2) Solubility of solid dispersions (F1-F6), (C) is a graph showing the elution profile of mebendazole solid dispersion (F5) in 0.1 M HCl medium containing 0.5% SDS (sodium dodecyl sulfate). Results are mean ± SD, (n=3), and all analyzes indicate statistical significance at P < 0.05 (*), P < 0.01 (**), and P < 0.001 (***).
Figure 3 is a graph showing the physicochemical properties of a solid dispersion containing mebendazole (MBZ). (A) XRD, (B) DSC, and (C) FTIR spectra for mebendazole, SDS, PM, and solid dispersion (F5), respectively.
Figure 4 is a microscopic view of the surface morphology of (A) mebendazole, (B) SDS, (C) PM, and (D) solid dispersion (F5).
Figure 5 is a graph showing the results of in vitro cytotoxicity evaluation of mebendazole (MBZ) and mebendazole-containing solid dispersion (F5) on cancer cells, (A) MDA-MB-231, (B) MCF-7. , (C) A549, (D) NCI-H1299, (E) HepG2, and (F) This is a graph showing the evaluation results in HeLa cells. Results are mean ± SD, (n=3), and all analyzes indicate statistical significance as P < 0.05 (*), P < 0.01 (**), and P < 0.001 (***)
Figure 6 is a graph of the plasma concentration profile after oral administration of 20 mg/kg of mebendazole (MBZ) and mebendazole-containing solid dispersion (F5) to rats. Results are mean ± SD, (n=3), and all analyzes indicate statistical significance at P < 0.05 (*), P < 0.01 (**), and P < 0.001 (***).

Hereinafter, preferred embodiments of the present invention will be described in detail. However, the present invention is not limited to the embodiments described herein and may be embodied in other forms. Rather, the content introduced herein is provided to be thorough and complete, and to fully convey the spirit of the invention to those skilled in the art.

Example One. of mebendazole Is it acceptable? High carrier selection experiment

1% (w/v) carrier [PVP (polyvinylpyrrolidone), Urea, Poloxamer 188 (pol188), Poloxamer 407 (pol407), Polyethylene glycol 6000 (PEG6000), Hydroxypropyl cellulose (HPC), Hydroxypropyl-β-cyclodextrin (HP-β) -CD), Hydroxypropyl methylcellulose (HPMC), Sodium dodecyl sulfate (SDS), Polyvinyl alcohol (PVA), and mannitol] by adding an excess amount of mebendazole to 1 ml of distilled water aqueous solution containing give. The mixture was mixed in a vortex for 1 min and incubated for 24 hours at 37 ± 0.5 °C in an incubator (LSI-3016A, Daihan Lab Tech Co., Ltd, Namyangju, Korea). After incubation, samples are filtered using a 0.45 μM nylon syringe filter (Whatman, International Ltd., Maidstone, UK). Mebendazole solubility was measured using high performance liquid chromatography (HPLC) with a CAPCELL PAK C18 column (150 × 4.6 mm, 5-μm particle size, OSAKA SODA). The mobile phase contained phases A and B in a ratio of 50:50 (v/v). Phase A consists of methanol and acetonitrile (AcCN) in a ratio of 3:2 (v/v), respectively. Phase B is 0.05M monobasic potassium phosphate. The injection volume is 20 μL and the flow rate is 1 mL/min. Mebendazole was measured at a wavelength of 290 nm.

As shown in Figure 1, to select a suitable carrier for mebendazole solid dispersion, PVP, urea, pol188, pol407, PEG6000, HPC, HP-β-CD, HPMC, SDS, PVA, and mannitol were selected and tested. As a result, the water solubility of mebendazole was highest in 1% SDS (sodium dodecyl sulfate) aqueous solution (solubility, 29.7 ± 0.34 μg/mL), so SDS was selected as the carrier to prepare a solid dispersion formulation.

Example 2. of mebendazole solid dispersion formulation manufacturing

Mebendazole solid dispersion is made by mixing the drug and SDS (Sodium dodecyl sulfate) carrier in the ratio of 1:1 (F1), 1:2 (F2), 1:3 (F3), 1:5 (F4), and 1:7 ( F5), and 1:10 (F6) (w/w) were mixed and then lyophilized. That is, mebendazole is dissolved in formic acid, the carrier (SDS) is dissolved in distilled water, and then the mebendazole solution is added dropwise to the carrier (SDS) solution while stirring at 500 rpm to prepare a mixed solution. Afterwards, the mixed solution was frozen at -70°C and then lyophilized using a freeze dryer (FDU-1200 EYELA, Tokyo Rikaakikai Co., Ltd, Tokyo, Japan) to obtain a mebendazole solid dispersion.

Experiment example One. of mebendazole Water solubility and in vitro dissolution test

The saturated solubility of the solid dispersion was measured in distilled water and 0.1 M HCl media. Excess of the solid dispersion was placed into a microcentrifuge tube containing 1 mL distilled water and 0.1 M HCl. The mixture was mixed for 1 minute and incubated at 37 ± 0.5°C for 24 hours. Afterwards, the mixture was centrifuged at 12,000 rpm for 10 minutes, and the supernatant was analyzed by HPLC as described above to measure the solubility of mebendazole in the solid dispersion formulation.

Dissolution studies were performed in acid medium (0.1 M HCl) at 37 ± 0.5°C according to the USP Apparatus II paddle method. The rotation speed was 75 rpm. Because the solubility of mebendazole is limited in acidic media, 0.5% sodium dodecyl sulfate (SDS) was added to 0.1 M HCl medium. Mebendazole pure drug (50 mg) and solid dispersion (equivalent to 50 mg of mebendazole) were added to 900 mL of medium. At appropriate time intervals of 5, 10, 15, 30, 45, 60, 90, and 120 min, samples were separated from the eluate and filtered using a 0.45 μm nylon syringe filter. Mebendazole concentration was measured by HPLC as previously described.

As shown in Figure 2(A), the water solubility of mebendazole solid dispersion in distilled water is highest in solid dispersion F5 (MBZ: SDS, 1:7, w/w; 4,614 ± 32.8 μg/mL), Next is F6 (MBZ: SDS, 1:10, w/w; 4,310 ± 30.1 μg/mL) and F4 (1:5, w/w; 2,348 ± 4.50 μg/mL). The water solubility of the mebendazole solid dispersion in Figure 2(B) in 0.1 M HCl medium showed the best water solubility in F5 (MBZ: SDS, 1:7) and F6 (MBZ: SDS, 1:10). and mebendazole solid dispersion F5 was selected and further experiments were conducted. In Figure 2(C), mebendazole solid dispersion (F5) shows significantly higher elution of mebendazole than mebendazole powder even in 0.1 M HCl medium containing 0.5% SDS (sodium dodecyl sulfate). Mebendazole solid dispersion (F5) shows 87.9 ± 0.55% of mebendazole elution within 5 minutes, which is 1.5 times higher than mebendazole dispersion, and mebendazole solid dispersion (F5) shows mebendazole dissolution within 30 minutes. It shows almost 100% mebendazole dissolution, which is expected to be the result of increased water solubility of mebendazole solid dispersion (F5).

Experiment example 2. Mebendazole solid dispersion PXRD , D.S.C. , and FTIR analyze

For comparison with the mebendazole solid dispersion (F5) formulation, it was a physical mixture (PM) with the same composition ratio as F5, that is, mebendazole drug and SDS at a weight ratio of 1:7 (w/w). A powder prepared by mixing in a mortar was used.

Structural characteristics of mebendazole, SDS, PM (Physical mixture), and mebendazole solid dispersion formulation (F5) were determined using powder X-ray diffraction (PXRD, X-Pert PRO MPD, Rigaku International Corp., Tokyo Japan). It was measured. Data from the samples were collected with a diffraction angle 2θ with an angular range consisting of between 5° and 50°. Differential scanning calorimetry (DSC) of mebendazole, SDS, PM (Physical mixture), and solid dispersion (F5) was performed at 10 °C using TGA/DSC1 (Mettler-Toledo International Inc., Greifensee, Switzerland). Measurements were made between 30 and 300 °C at a rate of /min. The possible interaction between mebendazole drug and polymer in solid dispersions was determined using Fourier-transform infrared spectroscopy (FTIR, Alpha-P, Brucker Optik, Ettligen, Germany). Measurements were made in the wavelength range of 500 to 4000 cm ^{- 1} .

Figure 3(A) shows the PXRD results of mebendazole, SDS, PM (Physical mixture), and solid dispersion (F5). The 2θ values of mebendazole crystals are 12.2, 14.4, 16.3, 17.2, 18.2, 19.7, 21.4, 23.4, 24.7, 26.8, 28.6, 29.1, and the 2θ values of SDS are 8.2, 9.0, 11.3, 13.7, 18.2, 20.3, 20.7, 21.8, 22.9, 27.5, 29.9, 32.3, 34.6 in PM samples. and There is an SDS peak, and in the solid dispersion (F5), a peak shift is observed due to the interaction between mebendazole and SDS, which causes the increase in solubility and elution of mebendazole in the solid dispersion (F5). It is confirmed that Figure 3(B) shows the DSC results of mebendazole, SDS, PM (Physical mixture), and solid dispersion (F5). The melting point of mebendazole is 258°C, and the melting point of SDS is 198°C. In the solid dispersion (F5), the melting points of mebendazole and SDS disappear, indicating that there is an interaction between the mebendazole drug and the carrier SDS. Figure 3(C) shows the FTIR results of mebendazole, SDS, PM (Physical mixture), and solid dispersion (F5). The shift from the original peak positions of mebendazole and SDS in the solid dispersion (F5) is mebendazole. It represents the interaction between and SDS.

Experiment example 3. Mebendazole solid dispersion SEM analyze

The sample is fixed in a sample holder and coated with gold for 1 minute. The surface morphology of mebendazole (MBZ), SDS, PM (Physical mixture), and solid dispersion (F5) was examined using Field Emission Scanning Electron Microscopy (FE-SEM III Merlin Compact, Carl Zeiss, Oberkochen, Germany) was used to measure it. The analysis voltage is 10 kV.

Figure 4 shows the SEM results of mebendazole (A), SDS (B), PM (Physical mixture) (C), and solid dispersion (F5) (D), showing mebendazole solid dispersion (D) having a smooth surface. ) SEM shows the interaction between mebendazole and SDS.

Experiment example 4. In vitro anticancer effect experiment

Cytotoxicity of mebendazole against various cancer cells, such as breast cancer cells (MDA-MB-231 and MCF-7), lung cancer cells (NCI-H1299 and A549), liver cancer cells (HepG2), and cervical epithelial cancer cells (HeLa). The in vitro anticancer effect was measured. MDA-MB-231, MCF-7, NCI-H1299, and A549 cells were cultured in RPMI medium containing 10% (v/v) FBS and 1% (v/v) antibiotics (penicillin/streptomycin) with 5% CO ₂ and cultured at 37 ± 0.5°C. Cells are seeded in 96-well plates at a concentration of 1 × 10 ⁴ cells per well in complete medium conditions. Once cell attachment occurs, cells are placed in new medium containing mebendazole alone and mebendazole solid dispersion (F5) at concentrations of 0.5, 1, 2, 5, 10, 20, and 30 μg/mL mebendazole. It is let go. After culturing at 37 ± 0.5°C for 24 hours, remove the medium and add 100 μL of MTT solution (0.5 mg/mL). After culturing for another 3 hours, the supernatant was removed and 100 μL of DMSO was added to dissolve the purple formazan crystals. Sample absorbance was measured using a microplate reader (Infinite M200 PRO; Tecan Trading AG, M nnedorf, Switzerland) and measured at 570 nm. Cell viability (%) is calculated using the formula below.

Figure 5 shows in vitro cytotoxicity results by MTT assay. Figure 5(A) shows the cell survival rate (%) of MDA-AB-231 cancer cells, Figure 5(B) shows the cell survival rate (%) of MCF-7 cancer cells, and Figure 5(C) shows the cell survival rate (%) of A549 cancer cells. , Figure 5(D) shows the cell survival rate (%) of NCI-H1299 cancer cells, Figure 5(E) shows the cell survival rate (%) of HepG2 cancer cells, and Figure 5(F) shows the cell survival rate (%) of HeLa cancer cells. . At 30 μg/mL, mebendazole solid dispersion (F5) compared to mebendazole in these cancer cells (breast cancer cells: MDA-MB-231 and MCF-7; lung cancer cells: NCI-H1299 and A549; liver cancer cells: It shows about 7 times stronger cytotoxicity against HepG2; cervical epithelial cancer cells: HeLa). Table 1 below shows the IC ₅₀ (Half maximal inhibitory concentration) values of mebendazole and mebendazole solid dispersion (F5) against cancer cells using GraphPad Prism 5 software (San Diego, CA, USA). am.

IC ₅₀ of mebendazole and mebendazole solid dispersion (F5) against cancer cells cell line IC ₅₀ (μg/mL) (mean ± SD, 95% CI) Mebendazole (MBZ) Mebendazole solid dispersion (F5) MDA-MB-231 29.9 ± 1.06 (26.3-34.0) 6.57 ± 1.09 (5.40-7.98)*** MCF-7 12.2 ± 1.10 (9.93-15.0) 3.88 ± 1.14 (2.96-5.10)*** NCI-H1299 6.69 ± 1.06 (5.95-7.53) 2.43 ± 1.08 (2.05-2.87)*** A549 44.9 ± 1.12 (35.7-56.6) 12.6 ± 1.19 (8.81-18.0)*** Hela 6.22 ± 1.09 (5.15-7.51) 1.83 ± 1.19 (1.27-2.64)*** HepG2 26.3 ± 1.06 (23.0-29.9) 5.03 ± 1.11 (4.05-6.26)*** * P < 0.05, ** P < 0.01, and *** P < 0.001

Experiment example 5. Pharmacokinetics ( Pharmacokinetics ) Experiment

Pharmacokinetic experiments were conducted according to the protocol approved by the Ethics Committee of the Chungnam National University, South Korea (No. 202003A-CNU-057). Male Sprague-Dawley rats (220 ± 10 g) were obtained from Samtako Bio Korea. Animals were housed at 22 ± 2°C, relative humidity of 50-60%, and 12-hour light/dark cycle per day, and food and water were provided ad libitum to the rats. After a week, the animals were divided into two groups for the experiment. After starving for a day, mebendazole and mebendazole solid dispersion (F5) are orally administered at a dose of 20 mg/kg (3 animals in each group). Blood samples from rats are collected by posterior orbital plexus every 0, 0.5, 1, 2, 4, 8, 12, and 24 hours and stored in heparin-containing tubes. Plasma samples are obtained by centrifuging blood samples at 12,000 rpm for 10 minutes in a centrifuge (Hanil micro-12, Hanil Scientific Inc., Gimpo, Korea), and the obtained plasma samples are stored at -70°C before analysis.

For analysis, add 500 μL of AcCN to a microcentrifuge tube containing 100 μL plasma sample and vortex mix for 2 minutes for protein precipitation. The sample is then centrifuged at 12,000 rpm for 10 minutes to separate the precipitated proteins. The supernatant was placed in a new tube and evaporated in a vacuum desiccator. The dried residue was added to 100 μL mobile phase for reconstitution and vortex mixed for 1 minute. Centrifuge the sample at 12,000 rpm for 10 minutes and inject 20 μL sample into the HPLC system as previously mentioned.

Pharmacokinetic parameters of mebendazole and mebendazole solid dispersion were calculated using the non-compartmental model of the WinNonLin program. C _max and the time to reach C _max (T _max ) are obtained directly from the plasma concentration curve. The area under the curve (AUC) is calculated according to the linear trapezoidal rule (a method of calculating the area by assuming that the area between the point where the concentration is measured on the concentration-time curve is a trapezoid). The terminal half-life (T _1/2 ) is calculated as 0.693/ λ _z , where λ _z (K _el ) is the terminal elimination rate constant calculated by log concentration linear regression. )am.

Figure 6 shows the plasma concentration profile of mebendazole after oral administration of mebendazole to rats. Mebendazole solid dispersion (F5) shows superior mebendazole plasma concentration at all points compared to mebendazole administration alone. Drug absorption depends on the rate and extent of drug absorption and is determined by several factors such as solubility, permeability, pKa, or lipophilicity. In the present invention, mebendazole is a hydrophobic drug and poorly soluble drug corresponding to BCS Class II. By manufacturing it with a selected SDS carrier and mebendazole solid dispersion, bioavailability was maximized by increasing solubility and dissolution rate. Table 2 shows the pharmacokinetic parameters after oral administration of mebendazole powder and mebendazole solid dispersion (F5) to rats (20 mg/kg). Mebendazole solid dispersion (F5) is mebendazole powder. It shows excellent bioavailability about 4 times higher than that of

Rat pharmacokinetic parameters of mebendazole and mebendazole solid dispersion (F5) Parameters MBZ SD (F5) AUC _{0 -24} (μg·h·mL ^-1 ) 32.3 ± 0.54 115.1 ± 1.44*** AUC _{0 -∞} (μg·h·mL ^-1 ) 33.8 ± 0.52 136.0 ± 1.25*** C _max (μg·mL ^-1 ) 2.20 ± 0.14 7.28 ± 0.23*** T _max (h) 8.00 ± 0.00 2.00 ± 0.00 T _1/2 (h) 4.66 ± 0.14 9.49 ± 1.27** K _el (h ^-1 ) 0.15 ± 0.00 0.07 ± 0.01*** MRT _last (h) 9.54 ± 0.04 9.59 ± 0.20 Relative bioavailability (%) 100 402*** * P < 0.05, ** P < 0.01, *** P < 0.001, AUC: area under the curve, T _1/2 : half-life, C _max : maximum plasma concentration, T _max : the time taken to reach maximum concentration, K _el : elimination rate constant, MRT: mean residence time.

Experiment example 6. Statistical analysis

Data are expressed as mean ± standard deviation (SD). One-way analysis of variance (ANOVA) in SigmaPlot 12.5 (SYSTAT Software, Inc., San Jose, CA, USA) was used for statistical analysis. P < 0.05 (*), P < 0.01 (**), and P < 0.001 (***) indicate significant differences.

Claims (7)

A solid dispersion containing mebendazole with improved bioavailability, comprising mebendazole and sodium dodecyl sulfate (SDS), wherein mebendazole in the solid dispersion exists in an amorphous form, and mebendazole : Mebendazole-containing solid dispersion, characterized in that the weight ratio of sodium dodecyl sulfate is 1:7~10 (w/w). delete delete delete delete An anticancer composition comprising the mebendazole-containing solid dispersion according to claim 1. A method for producing a mebendazole-containing solid dispersion according to claim 1,
a) dissolving mebendazole in formic acid to prepare a mebendazole solution, and dissolving sodium dodecyl sulfate in distilled water to prepare a sodium dodecyl sulfate solution;
b) adding a mebendazole solution to the sodium dodecyl sulfate solution and stirring to prepare a mixed solution; and
c) freeze-drying the mixed solution; a method for producing a solid dispersion containing mebendazole, comprising: