KR101564926B1 - Pharmaceutical preparation for treating chronic myelogenous leukemia containing yellow-poplar bark extract as active ingredient - Google Patents

Abstract

The present invention relates to a pharmaceutical preparation for the treatment of chronic myelogenous leukemia comprising an extract of Liliaceae bark as an active ingredient. More specifically, in order to selectively inhibit the mutant enzyme T315I caused by the mutation of the BCR-ABL fusion gene, which is a causative gene of chronic myelogenous leukemia, poly ADP ribosyl polymerization associated with mitochondrial membrane potential (MMP) The present invention relates to a medicament for the treatment of chronic myelogenous leukemia containing an extract of lily bark extract which causes apoptosis through cleavage of the enzyme (PARP) and caspase-3.

Description

[0001] The present invention relates to a pharmaceutical preparation for treating chronic myelogenous leukemia comprising an extract of Liliaceae bark as an active ingredient,

The present invention relates to a pharmaceutical preparation for the treatment of chronic myelogenous leukemia comprising an extract of Liliaceae bark as an active ingredient. More specifically, in order to selectively inhibit the mutant enzyme T315I caused by the mutation of the BCR-ABL fusion gene, which is a causative gene of chronic myelogenous leukemia, poly ADP ribosyl polymerization associated with mitochondrial membrane potential (MMP) The present invention relates to a medicament for the treatment of chronic myelogenous leukemia containing an extract of lily bark extract which causes apoptosis through cleavage of the enzyme (PARP) and caspase-3.

Liliodendron tulipifera L. (Yellow-poplar) is a deciduous broad-leaved arboreous tree belonging to Magnoliaceae. It has been reported that various compounds such as alkaloids, sesquiterpenes and lignan trees are contained in bark extracts of lily trees .

The extracts of Liliaceae bark extract include costunolide, tulipinolide, epi-tulipinolide, epi-tulipdienolide and gamma-llyiodonolide. (Gamma-liriodenolide) and the like.

Among these sesquiterpinolactones, the phenolonide has been identified as an anti-inflammatory or anticancer effect mediated by the nuclear transcription factors NF-κB, p21 and cyclin D1, and in particular, LC- 1 (dimethylamino-parthenolide) has been reported to induce anticancer effects on acute myeloid leukemia (AML) by inducing cellular proliferation through inhibition of nuclear transcription factor NF-κB activity.

Leukemia is a type of hematologic malignancy in which blood cells, especially white blood cells, grow abnormally. White blood cells that do not mature properly are present in large quantities in the blood. Compared to the abnormal proliferation of leukocytes, the number of normal blood cells becomes extremely small, which makes it impossible to perform basic blood functions such as oxygen transport and nutrition, as well as immune function. Abnormal white blood cells also cause a similar reaction to autoimmune diseases It also destroys normal tissues.

Chronic myelogenous leukemia is a myeloproliferative disorder in which myeloid leukemia cells in the bone marrow grow uncontrolled and accumulate in the blood. The types of leukemia are divided into acute myelogenous leukemia, chronic myelogenous leukemia, acute lymphocytic leukemia (ALL), and chronic lymphocytic leukemia (CLL).

Outbreaks of chronic myelogenous leukemia In general, abnormal myeloid cells appear mature but do not function properly. Such chronic myelogenous leukemia is associated with an abnormal intracellular genetic marker called the " Philadelphia chromosome ".

Chronic myelogenous leukemia differs from other common leukemias, in particular the transition from the chronic multiplication stage of slowly progressive abnormal cells to the acute stage of abrupt proliferation. Chronic myelogenous leukemia may remain chronic for 3-4 years, and treatment at this time is more effective than inhibiting the disease. However, most of the patients with sudden switchover are very difficult to treat.

Chronic myelogenous leukemia is caused by the BCR-ABL fusion gene, the Philadelphia chromosome, in which the ABL gene on chromosome 9 and the BCR gene on chromosome 22 are translocated. Such a BCR-ABL fusion gene is involved in the biosynthesis of the carcinogenesis protein p210 by expressing a fusion protein having tyrosine kinase activity, and the biosynthetic p210 protein inhibits the cell progenitor cells of the bone marrow precursor cell, Leading to myeloproliferative disorders due to proliferation.

Imatinib (Gleevec) is a tyrosine kinase inhibitor of the first generation, BCR-ABL. It has been used for a long time as a first-line treatment for myeloid leukemia. However, patients with imatinib-resistant patients . Fortunately, second-generation treatments that overcome imatinib resistance, such as Dasatinib, Nilotinib, and Bosutinib, have been developed and are expected to have more efficacious and diverse drug treatments, T315I mutant 'mutant' ABL mutant 'T315I' mutations in the first generation and second generation drugs cause these second generation drugs to have no therapeutic effect. It is urgent to develop a third-generation therapeutic agent that is effective in suppressing the mutated gene.

For this purpose, the present inventors have tried to separate and purify the pharmacologically active components such as epi-tulipyinolide and cosolinide from the bark of lilia, and then to evaluate the therapeutic effect of each active ingredient on chronic myelogenous leukemia, And a Korean patent application No. 10-2013-46506 entitled " Pharmaceutical composition for treating chronic myelogenous leukemia containing lily extracts "

However, the bark extract of Liliaceae is insoluble, and when it is administered to a living body, its solubility and dissolution rate in the digestive juice are low in the process of being absorbed into the body, so that bioavailability can be lowered. Therefore, various formulation means should be developed to improve solubilization or dissolution rate of poorly soluble drugs including lily bark extract.

Accordingly, the present inventors have continued the formulation studies to maximize the therapeutic effect and minimize side effects in using the extract of Liliaceae bark as a therapeutic agent for chronic myelogenous leukemia. As a result, the present inventors have found that a pharmaceutical formulation showing excellent therapeutic effect on chronic myelogenous leukemia And the present invention has been completed by inventing a medicinal preparation capable of improving the solubilization or dissolution rate of the hardy lily bark extract.

The object of the present invention is to develop a medicinal formulation for maximizing the therapeutic effect and minimizing adverse effects in using lily bark extract as a therapeutic agent for chronic myelogenous leukemia. And to develop a pharmaceutical preparation which can improve the solubilization or dissolution rate of the hardy lily bark extract.

It is an object of the present invention to provide a process for the production of polyoxyethylene sorbitan fatty acid esters, polyoxyethylene sorbitan fatty acid esters, polyoxyethylene fatty acid esters, polyoxyethylene Polyoxypropylene block copolymers, polyoxyethylene polyoxypropylene copolymers, reaction products of natural or hydrogenated vegetable oils with ethylene glycol, sodium dioctylsulfosuccinate, sodium laurylsulfonate, phospholipids, propylene glycol fatty acid esters, natural vegetable oils 5 to 100 parts by weight of a transesterification reaction product of triglyceride and polyalkylene polyol, at least one surfactant selected from mono-glyceride, di-glyceride or sterol; And ii) 50 to 400 parts by weight of at least one co-solvent selected from the group consisting of propylene carbonate, propylene glycol, ethanol, diethylene glycol monoethyl ether, glycopyrrole, polyethylene glycol, dimethylisosorbide and N-methylpyrrolidone; Lt; RTI ID = 0.0 > T315I < / RTI > due to a BCR-ABL fusion gene comprising the BCR-ABL fusion gene.

Wherein the non-aqueous medicinal preparation comprises 10 to 50 parts by weight of a surfactant and 100 to 200 parts by weight of a co-solvent based on 100 parts by weight of the extract of Liliaceae bark.

The surfactant may be at least one selected from the group consisting of polyoxyethylene sorbitan fatty acid esters, sorbitan fatty acid esters, polyoxyethylene fatty acid esters, polyoxyethylene polyoxypropylene block copolymers and polyoxyethylene polyoxypropylene copolymers .

The co-solvent may be at least one selected from the group consisting of propylene glycol, polyethylene glycol and diethylene glycol monoethyl ether.

Yet another object of the present invention is to provide a soft capsule which is useful for the treatment of chronic myelogenous leukemia comprising the non-aqueous medicament form as a content liquid and molded into a gelatin capsule.

The effect of the present invention is to provide a medicinal formulation for maximizing the therapeutic effect and minimizing adverse effects in using lily bark extract as a therapeutic agent for chronic myelogenous leukemia. And to provide a medicinal preparation capable of improving the solubilization or dissolution rate of the hardy lily bark extract.

BRIEF DESCRIPTION OF THE DRAWINGS Fig. 1 compares the dissolution rate over time of the soft capsule preparation (test preparation) and tablets (comparative preparation) of the present invention containing 30 mg of Lilium bark extract in the pH = 1.2 buffer solution of Example 2. Fig.
Figure 2 compares the dissolution rate over time of the soft capsule preparation (test formulation) and tablets (comparison formulation) of the present invention containing 30 mg of Lilium bark extract in the pH = 6.8 buffer solution of Example 2.

The present invention relates to 100 parts by weight of an extract of Liliaceae bark extract containing epi-tulipinolide as an active ingredient, i) a polyoxyethylene sorbitan fatty acid ester, a sorbitan fatty acid ester, a polyoxyethylene fatty acid ester, a polyoxyethylene polyoxy Propylene block copolymers, polyoxyethylene polyoxypropylene copolymers, reaction products of natural or hydrogenated vegetable oils with ethylene glycol, sodium dioctylsulfosuccinate, sodium laurylsulfonate, phospholipids, propylene glycol fatty acid esters, natural vegetable oil triglycerides And 5 to 100 parts by weight of at least one surfactant selected from mono-glycerides, di-glycerides and sterols; And ii) 50 to 400 parts by weight of at least one co-solvent selected from the group consisting of propylene carbonate, propylene glycol, ethanol, diethylene glycol monoethyl ether, glycopyrrole, polyethylene glycol, dimethylisosorbide and N-methylpyrrolidone; Lt; RTI ID = 0.0 > T315I < / RTI > due to the BCR-ABL fusion gene comprising the BCR-ABL fusion gene.

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

The non-aqueous medicinal preparation of the present invention comprises 10 to 50 parts by weight of a surfactant and 100 to 200 parts by weight of a co-solvent based on 100 parts by weight of the extract of Liliaceae bark.

The surfactant is at least one selected from polyoxyethylene sorbitan fatty acid esters, sorbitan fatty acid esters, polyoxyethylene fatty acid esters, polyoxyethylene polyoxypropylene block copolymers and polyoxyethylene polyoxypropylene copolymers.

The co-solvent is at least one selected from propylene glycol, polyethylene glycol, and diethylene glycol monoethyl ether.

The present invention also provides a soft capsule which is useful for the treatment of chronic myelogenous leukemia comprising the non-aqueous pharmaceutical preparation as a content liquid and formed into a gelatin capsule.

The present invention provides a pharmaceutical composition of a lily extract useful for selectively inhibiting T315I, which is a mutant enzyme caused by a BCR-ABL fusion gene which is a causative gene of chronic myelogenous leukemia, and epi-tulipipinoide, which is an active ingredient thereof, The active ingredient may have one or more known active ingredients having the same effects as the pharmaceutical composition or extract of the present invention.

In the present invention, as another medicinal preparation other than soft capsules, tablets, pills, powders, hard capsules, gelatin-billed hard capsules, caramel or jelly-type chewing tablets and oral A solution for oral use selected from a solution for oral administration can be provided.

In addition, the bark bark extract of the present invention may further comprise, as an active ingredient, Epitulipdienolide, Ridentin or Deacetyllipiperolide.

The non-aqueous pharmaceutical formulations of the present invention may be administered in the form of tablets, granules, capsules, powders, syrups, ointments, suppositories and subcutaneous, intramuscular, intravenous or dermal injections as well as soft capsules, 10 to 500 mg per day can be taken in one to several divided doses.

In addition, the composition may increase the solubility and absorption in the gastrointestinal tract of a substance commonly used in pharmaceuticals, for example, extracts of lily bark, and may be dispersed and emulsified with water at the time of oral administration to such an extent as not to adversely affect drug efficacy, Additives such as acids, fatty acids or fatty acid alcohols which can be widely used for increasing the bioavailability, saccharides such as saccharides such as white sugar, MacIion sugar, white sugar, gelatin, sugar and starch syrup, lubricants such as magnesium stearate, talc, An antioxidant, a flavoring agent, an antiseptic, a fragrance, a sweetener, a coloring agent, a pH adjusting agent and a viscosity adjusting agent may be further added to the composition, such as cellulose, anhydrous calcium chloride, starch, excipients such as mannitol, It is preferable to add the extract in an amount usually used for the extract of wood bark.

The acids, fatty acids or fatty acid alcohols that can be used in the composition of the present invention include citric acid, oleic acid, stearyl alcohol, myristic acid, linoleic acid or lauric acid, capric acid, caprylic acid, caproic acid ), But the present invention is not limited thereto.

Antioxidants that may be used in the compositions of the present invention include butylated hydroxytoluene, sodium bisulfite,? -Tocopherol, vitamin C,? -Carotene, tocopherol acetate, fumaric acid, nalic acid, Butylated hydroxyanisole, propyl gallate, sodium ascorbate, and the like can be used, but the present invention is not limited thereto.

Flavoring agents that can be used in the composition of the present invention include, but are not limited to, a mixed fruit, an apple, a strawberry, a cherry, a pepper, a vanilla, a yogurt or a drink.

Preservatives which can be used in the composition of the present invention include, but are not limited to, benzoic acid, sodium benzoate, ethylparaben, methylparaben or propylparaben.

Examples of the fragrance that can be used in the composition of the present invention include, but are not limited to, peppermint leaves, peppermint oil, orange oil, clove oil, cinnamon oil, strawberry essence and other common fruit or vegetable essences.

Examples of the sweetening agent that can be used in the composition of the present invention include, but are not limited to, black sugar, glucose, fructose, aspartame, stevioside, sorbitol, mannitol, oligosaccharide,

The coloring matters which can be used in the composition of the present invention include green 3, red 2, red 3, blue 1, blue 2, yellow 4, yellow 5, water-soluble mannitol, caramel, titanium oxide, But is not limited thereto.

The pH adjusting agent which can be used in the composition of the present invention may be sodium carbonate, sodium hydroxide, potassium hydroxide, triethanolamine or monoethanolamine, but is not limited thereto.

Viscosity modifiers that can be used in the compositions of the present invention include, but are not limited to, hydroxypropylcellulose (HPC), hydroxypropylmethylcellulose (HPMC), hydroxyethylcellulose, ethylcellulose, methylcellulose, carboxymethylcellulose (CMC), acacia, Bentonite, alginic acid, propylene glycol alginate, polyvinylpyrrolidone, polyvinyl alcohol (PVC), carbopol, polycarbopil, tragacanth or xanthan gum ), But the present invention is not limited thereto.

Further, in addition to the above-mentioned active ingredients, pharmaceutically acceptable carriers, excipients and diluents may be further included for administration of the pharmaceutical composition of the present invention.

Examples of carriers, excipients and diluents that can be included in the solution include lactose, textolose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose , Microcrystalline cellulose, polyvinylpyrrolidone, purified water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, and mineral oil.

Solid formulations for oral administration include tablets, pills, powders, granules, capsules, hard capsules, soft capsules, caramel or jelly type chewing tablets, etc. These solid preparations may contain at least one excipient , Such as starch, calcium carbonate, sucrose or lactose, gelatin, and the like. In addition, lubricants such as magnesium stearate and talc may be used.

Liquid preparations for oral administration include suspensions, solutions, emulsions, syrups and the like. In addition to the commonly used simple diluents such as purified water and liquid paraffin, various excipients such as wetting agents, sweetening agents, have.

Examples of the preparation for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, freeze-dried preparations, suppositories and the like. Non-aqueous solvents and suspensions include vegetable oils such as propylene glycol, polyethylene glycol and olive oil, An injectable ester such as a rate can be used. Examples of the suppository base include withexol, macrogol, tween 61, cacao butter, laurin, glycerogelatin and the like.

The extract of Liliaceae bark and its pharmaceutical composition of the present invention may be administered in various routes according to the intended method in addition to the oral administration, and may be administered orally, rectally, intravenously, muscularly, subcutaneously, intraperitoneally, .

Hereinafter, the present invention will be described in detail with reference to examples. However, the following examples illustrate the present invention and the scope of the present invention is not limited by the following examples.

(Production Example 1) Separation and preparation of lily bark extract (CD-200)

1. Solvent Extraction Process

Three to five years old Lilium bark (200 g) was shaken with 2000 ml of ethyl acetate at room temperature for 24 hours, and the extracted filtrate was concentrated under reduced pressure to obtain an ethyl acetate primary extract. 500 ml of butanol was added to the obtained primary extract After 2 to 4 hours of extraction at room temperature, the butanol layer was separated and removed to remove components dissolved in butanol, and then concentrated under reduced pressure to obtain 12.54 g of crude extract.

2. Refining Process

To the resulting crude extract (12.54 g) was dissolved 200 ml of 70% ethanol, 200 ml of n-hexane was added thereto and the mixture was shaken to obtain a 70% ethanol layer. The ethanol layer was separated and concentrated under reduced pressure to obtain 2.75 g of an extract Respectively. The extract contains epi-tulipinolide as a main component and is referred to as CD-200.

(Experimental Example 1) mitochondria-dependent apoptosis of leukemia cells by CD-200

CD-200 extract induces apoptosis of MMP (mitochondrial membrane potential) against T315I, a mutation of the BCR-ABL fusion gene, a gene responsible for chronic myelogenous leukemia, to induce cell proliferation and inhibit leukemia mutant cell proliferation . Specifically, CD-200 was treated with 1, 5 and 10 μg / ml of Bcr-Abl ^T315I Ba / F3 cell line (1 × 10 ⁶ cells / 10 ml) and incubated for 8 hours at 37 ° C. in a 5% CO ₂ incubator Respectively. After collecting the cells, 0.5 ml of JC-1 working solution (BD Biosciences) was added under the condition of blocking light, and incubated for 15 min at 37 ° C in a 5% CO ₂ incubator. In the group treated with BaF3 / T315I mutant at a concentration of 1 μg / ml, 5 μg / ml and 10 μg / ml for 6 hours, the nonpolarization was 9.42%, 18%, 77.9% % Concentration-dependent cell death (MMP depolarization). On the other hand, Imatinib (4.02%) showed a similar non-depolarization value to the control group (4.36%),

(Experimental Example 2) Caspase-3 activity-dependent apoptosis of leukemia cells by CD-200

CD-200 extracts were tested for T315I, a mutation of the BCR-ABL fusion gene, a gene responsible for chronic myelogenous leukemia, to induce caspase-3 activation and inhibit leukemia mutant gene cell proliferation. Specifically, CD-200 was treated with 1,5 μg / ml of Bcr-AblT315IBa / F3 cell line (8 × 10 ⁵ cells / 2 ml / well) and cultured in a 5% CO ₂ incubator at 37 ° C. for 12 hours. Cells were collected and incubated with Anti-Cleaved Caspase-3 (1: 800, Cell Signaling Technology, Beverly, Mass., USA) antibody and the corresponding iso-type Rabbit (DA1E) mAb IgG XP ㄾ Isotype Control Signaling Technology, Beverly, Mass., USA) was reacted for 30 minutes, and Green rabbit-FITC fluorescence was reacted for 20 minutes and caspase-3 activity was analyzed using a flow cytometer. As a result, Of CD-200, but no change was observed in Imatinib when compared to CD-200.

(Experimental Example 3) Western blot for confirmation of cell death of CD-200

Western blot analysis was performed to confirm whether the expression of PARP or Caspage-3, which is a cell death marker protein, was changed by using CD-200 in order to confirm whether Bcr-Abl ^T315I Ba / F3 cell line induces apoptosis.

To confirm the apoptotic pathway of CD-200, CD-200 was treated with 1 ~ 10 μg / ml of Bcr-Abl ^T315I Ba / F3 cell line (2 × 10 ⁶ / , And cultured in a 5% CO ₂ incubator. The Western blot experiment was carried out as follows. 30 μg of protein was added to 10% and 15% SDS polyacrylamide gels, electrophoresed for 2 hours, transferred to full rovinylidene fluoride (PVDF) and incubated with 5% non-fat skim milk for 1 hour (1: 3000, Cell Signaling Technology, Beverly, Mass., USA) and incubated overnight at 4 ° C. And the corresponding anti-rabbit IgG horseradish peroxidase-conjugate was treated and detected with ECL.

It was confirmed that BCD-200 induces apoptosis in Bcr-Abl ^T315I Ba / F3 cell line, unlike Imatinib, by forming a truncation form of PARP, Caspase-3, which is involved in apoptosis pathway at 5 μg / ml.

(Production Example 2) A soft capsule preparation containing lily bark extract (CD-200)

100 mg of bark extract of Liliaceae obtained in Preparation Example 1, 30 mg of polyoxyethylene sorbitan fatty acid ester and 20 mg of Clemophora EL were mixed. 150 mg of diethylene glycol monoethyl ether was added to the mixed solution, and the mixture was heated at 80 캜 to dissolve the mixture until it became transparent to prepare a composition. The composition was filled into soft capsules and formulated.

(Production Example 3) A soft capsule preparation containing lily bark extract (CD-200)

100 mg of bark extract of lily obtained in Preparation Example 1 and 40 mg of polyoxyethylene sorbitan fatty acid ester were mixed. 20 mg of Kremophor RH40 and 200 mg of diethylene glycol monoethyl ether were added to the mixed solution, and the mixture was heated at 80 DEG C to dissolve until the composition was transparent. The composition was filled into soft capsules and formulated.

(Production Example 4) A soft capsule preparation containing lily bark extract (CD-200)

100 mg of bark extract of lily obtained in Preparation Example 1, 30 mg of polyoxyethylene sorbitan fatty acid ester and 10 mg of polyoxyethylene polyoxypropylene block copolymer were mixed. 20 mg of Kremophor RH40 and 150 mg of diethylene glycol monoethyl ether were added to the mixed solution, and the mixture was heated at 80 占 폚 to dissolve until transparent to prepare a composition. The composition was filled into soft capsules and formulated.

(Preparation Comparative Example 1) Oral tablet containing lily bark extract (CD-200)

100 mg of the extract of Liliaceae bark extract obtained in Preparation Example 1 and 30 mg of Kremopo EL were mixed and mixed with 80 mg of colloidal silicone dioxide (Aerosil 200), followed by mixing and drying. Then, 20 mg of povidone, 80 mg of microcrystalline cellulose and 2 mg of magnesium stearate were mixed And tabletted to obtain tablets.

(Preparation Comparative Example 2) Oral tablet containing lily bark extract (CD-200)

100 mg of the extract of Liliaceae bark extract obtained in Preparation Example 1 and 30 mg of Kremopo EL were mixed and mixed with 70 mg of colloidal silicone dioxide (Aerosil 200), followed by mixing and drying. Then, 50 mg of povidone, 50 mg of microcrystalline cellulose and 2 mg of magnesium stearate were mixed And tabletted to obtain tablets.

(Example 1) Solubility comparison experiment

The solubility of the soft capsules of Lilium bark extract CD-200 by the automatic micellar drug delivery system of the present invention and the oral tablets of Lilium bark extract CD-200 without such a system were measured and compared.

Concretely, the solubility of the drug was measured in the pH condition (pH = 1.2) in the gastrointestinal tract and the pH condition (pH = 6.8) in the small intestine and large intestine with the contents of the preparation example 2 and the preparation before the preparation .

The experimental method is as follows.

An amount corresponding to 60 mg of the extract of Liliaceae bark extract was taken from the oral preparation prepared in Preparation Example 2 and Comparative Example 1, and a buffer solution of pH 1.2 (Korean Pharmacopoeia No. 8: 2.0 g of sodium chloride, 7.0 ml of hydrochloric acid, And a total volume of 1 L) and a buffer solution of pH 6.8 (manufactured by Kagaku Kogyo Co., Ltd., Amendment: 250 ml of 0.2 mol / L potassium dihydrogen phosphate solution, 118 ml of a 0.2 mol / L sodium hydroxide solution, 1 L) was added to the solution. After the solution was stirred at 37 占 폚 for 1 hour, the solubility was measured. Solubility was measured by UV after filtration. The analytical wavelength was 270 nm.

Solubility and comparison at pH = 1.2 and pH = 6.8 Formulation Solubility (/ / ml) pH 1.2 pH 6.8 Preparation Example 2 Content
(Test preparation) 400 400 Preparation Comparative Example 1 Mixture
(Comparative agent) 197 191 Solubility comparison
(Production Example 2 / Production Comparative Example 1) More than 2 times More than 2 times

As shown in Table 1, the solubilities of oral preparations (test preparations) prepared from Preparation Example 2 of the present invention were 400 μg / ml and 400 μg / ml at pH = 1.2 and pH = 6.8, The solubilities of the oral preparations (comparative preparations) prepared from Comparative Example 1 were respectively 197 μg / ml and 191 μg / ml. Comparing the above results, it can be seen that the test agent according to the present invention has a solubility two times higher than that of the comparative agent at pH = 1.2 and pH = 6.8, respectively. Therefore, the oral pharmaceutical preparation (test preparation) by the automatic micellar drug delivery system according to the present invention has a much higher solubility in the pH condition of the small intestine and the colon in the gastrointestinal tract than the pharmaceutical preparation prepared by the conventional method, It can be seen that it is uniformly dissolved even under the conditions.

(Example 2) Comparison of dissolution rate

The dissolution rates of the soft capsules containing the lily bark extract CD-200 by the automatic micellar drug delivery system of the present invention and the oral tablets of the lily bark extract CD-200 without such a system were measured and compared.

The experimental method is as follows.

The soft capsules prepared in Preparation Example 2 and the oral tablets prepared in Preparation Example 1 were mixed with 30 mg of the extract of lily bark extract, was added to 900 ml of buffer = pH = 6.8 and stirred at 50 rpm. Samples were taken at 5, 10, 15, 30, 45, 60, 90 and 120 min by paddle method and the contents of extracts of L. bark extract were measured. The dissolution rate was determined as% using the extract of Lark thunb. Bark extract, which was measured from the sample, as 100% of the extract of bark extract of lily contained in the oral preparation.

The dissolution rate measured at pH = 1.2 is shown in FIG. 1, and the dissolution rate measured at pH = 6.8 is shown in FIG.

As shown in Fig. 1, the soft capsule preparation (test preparation) prepared from the preparation example 2 of the present invention showed a dissolution rate of 60% or more in 10 minutes at pH = 1.2 and a dissolution rate of 70% or more in 15 minutes to 120 minutes And showed a constant dissolution rate. On the other hand, the oral tablets (comparative preparations) prepared from Comparative Preparation Example 1 showed a lower dissolution rate than the soft capsules prepared from Preparation Example 2. [ Therefore, it can be seen that the oral preparation containing the oral composition of the extract of Liliaceae bark by the automatic micellar drug delivery system according to the present invention has a high dissolution rate in the pH = 1.2 buffer solution.

2, the soft capsule preparation (test preparation) prepared from the preparation example 2 of the present invention showed a dissolution rate of 60% or more in 15 minutes at pH = 6.8 and a dissolution rate of 70% or more in 15 minutes to 60 minutes And showed a constant dissolution rate. On the other hand, the oral tablets (comparative preparations) prepared from Comparative Preparation Example 1 showed a lower dissolution rate than the soft capsules prepared from Preparation Example 2. [ Therefore, it can be seen that the oral preparation containing the oral composition of the extract of Liliaceae bark by the automatic micellar drug delivery system according to the present invention has a high dissolution rate in pH = 6.8 buffer solution.

As shown in FIG. 1 and FIG. 2, the oral preparation containing the oral composition of the extract of Liliaceae bark by the automatic micellar drug delivery system according to the present invention can be used as an oral composition for extracts of Liliaceae bark extract It can be seen that it has a high dissolution rate at pH = 1.2 and pH = 6.8 as compared with the oral preparation containing it, and it has a certain dissolution rate even at various pH.

Claims (5)

100 parts by weight of an extract of Liliaceae bark extract containing epi-tulipinolide as a main active ingredient, Epitulipdienolide, Ridentin or Deacetyllipiperolide as an auxiliary active ingredient, about
I) at least one surfactant selected from polyoxyethylene sorbitan fatty acid esters, sorbitan fatty acid esters, polyoxyethylene fatty acid esters, polyoxyethylene polyoxypropylene block copolymers or polyoxyethylene polyoxypropylene copolymers; Weight part; And
Ii) 50 to 400 parts by weight of at least one cosolvent selected from propylene carbonate, propylene glycol, ethanol, diethylene glycol monoethyl ether, glycopyrrole, polyethylene glycol, dimethylisosorbide or N-methylpyrrolidone;
Which is useful for the treatment of chronic myelogenous leukemia by selectively inhibiting the mutant enzyme T315I caused by the BCR-ABL fusion gene,
The non-aqueous pharmaceutical formulation according to claim 1, wherein the non-aqueous pharmaceutical formulation comprises 10 to 50 parts by weight of a surfactant and 100 to 200 parts by weight of a co-solvent, based on 100 parts by weight of the extract of Liliaceae bark.
delete The non-aqueous pharmaceutical preparation according to claim 1, wherein the co-solvent is at least one selected from the group consisting of propylene glycol, polyethylene glycol, and diethylene glycol monoethyl ether.
A soft capsule preparation useful for the treatment of chronic myelogenous leukemia comprising the non-aqueous pharmaceutical preparation of claim 1 as a liquid and formed into a gelatin capsule.

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