Controlled release composition for oral administration containing clay mineral complex, preparation method thereof and controlled release method

Abstract

The present invention relates to a controlled-release composition for oral administration and a preparation method thereof. The composition comprises a compound of Chemical Formula 1 or a pharmaceutically acceptable salt thereof and a complex of a clay mineral.

Description

Technical Field

[0001] The present invention relates to a controlled-release composition for oral administration containing a clay mineral complex, a preparation method thereof and a controlled-release method. Background Art

[0002] Smectite is a phyllosilicate mineral in which two octahedral sheets composed of Al, Mg, and Fe are sandwiched between tetrahedral sheets composed of Si, Al, and Fe to form a unit layer (2:1 layer). The unit layer of smectite has a negative charge, which is generated by the isomophic substitution of tetrahedral Si with a 4-valent positive charge by Al or Fe with a 3-valent positive charge, or the isomorphic substitution of octahedral Al or Fe3+ with a 3-valent positive charge by Mg or Fe2+ with a 2-valent positive charge. The negative charge generated in the unit layer induces cations between unit layers, which means that smectite can be used as a drug carrier.

[0003] The compound of the following chemical formula 1 is N-((4-([1,2,4]triazolo[1,5-a]pyridine-6-yl)-5-(6-methylpyridin-2-yl)-1H-imidazol-2-yl)methyl)-2-fluoroaniline recorded in Korean Patent No. 10-1500665, and is known to have the activity of existing anticancer agents. The present inventors confirmed the problem of irregular absorption of the compound in the body when studying the pharmacological effects of the above compound. Therefore, in the study to improve the in vivo effects of the compound of chemical formula 1, it was confirmed that when a complex is formed with clay minerals under specific conditions, not only the delivery of the drug to the colon can be improved, but also the drug effect in the colon can be improved, thereby completing the present invention.

[0004] [Chemical formula 1]

[0005] Summary of the invention

[0006] Technical issues

[0007] An object of the present invention is to provide a composition that improves drug delivery to the colon and drug action in the colon.

[0008] Technical Solution

[0009] In order to achieve the above object, the present invention provides a controlled-release composition for oral administration, which comprises a complex of a compound of Chemical Formula 1 or a pharmaceutically acceptable salt thereof and a clay mineral.

[0010] [Chemical formula 1]

[0011]

[0012] Furthermore, the present invention provides a method for preparing a controlled-release composition for oral administration, comprising the steps of mixing a solution containing a compound of Chemical Formula 1 or a pharmaceutically acceptable salt thereof, an acidic reaction solution, and a clay mineral suspension, and adsorbing the compound onto the clay mineral to prepare a complex, wherein the controlled-release composition for oral administration comprises a complex of the compound of Chemical Formula 1 and the clay mineral.

[0013] Furthermore, the present invention provides a controlled release method, characterized in that a composition is orally administered to a subject, wherein the pharmaceutical composition comprises a complex of the compound of Chemical Formula 1 or a pharmaceutically acceptable salt thereof and a clay mineral.

[0014] [Chemical formula 1]

[0015]

[0016] Beneficial Effects

[0017] The composition of the present invention can control the release of the compound of Chemical Formula 1, and can not only effectively deliver the compound to the colon, but also enable the compound to act slowly in the colon for a long time. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1 This is a picture taken by scanning electron microscope (SEM) after the surface of vactosertib powder was coated with platinum.

[0019] Figure 2 This is a scanning electron microscope (SEM) photograph of the bentonite / vactosertib composite after coating with platinum.

[0020] Figure 3 The graph shows XRD patterns of vactosertib, bentonite, a bentonite / vactosertib complex, and a mixture of vactosertib and bentonite simply mixed in the same ratio as the complex, after X-ray diffraction analysis (XRD) was performed.

[0021] Figure 4The drug release patterns of vactosertib powder (●) and bentonite / vactosertib complex (□) are shown when a 0.1 N HCl solution (pH 1.2) containing 0.1% Tween 80 was used as an eluent.

[0022] Figure 5 The drug release patterns of vactosertib powder (●) and bentonite / vactosertib complex (□) are shown when 50 mM acetate buffer (pH 4.5) containing 0.1% Tween 80 was used as an eluent.

[0023] Figure 6 The drug release patterns of vactosertib powder (●) and bentonite / vactosertib complex (□) are shown when 50 mM phosphate buffer (pH 7.4) containing 0.1% Tween 80 was used as an eluent.

[0024] Figure 7 The blood drug concentration curves over time after orally administering vactosertib powder (●), vactosertib aqueous solution (△) and bentonite / vactosertib complex suspension (□) to rats at a dose of 10 mg / kg based on the amount of vactosertib are shown.

[0025] Figure 8 Shown is the drug administration regimen using the DSS-induced ulcerative colitis rat model.

[0026] Fig. 9 The figure shows the change in the blood concentration of vactosertib over time in the DSS-induced ulcerative colitis rat model. The points connecting the curves represent the average blood concentration data of animals belonging to each group at different time periods, ○ represents the G1 group, ◇ represents the G2 group, ● represents the G3 group, and ◆ represents the G4 group.

[0027] Fig.10 The peak plasma concentration time (t) of vactosertib after oral administration of the test substance in the DSS-induced ulcerative colitis rat model is shown. max ), ○ represents G1 group, ◇ represents G2 group, ● represents G3 group, and ◆ represents G4 group.

[0028] Fig.11 The peak concentration (C) of vactosertib after oral administration of the test substance in the DSS-induced ulcerative colitis rat model is shown. max ), ○ represents G1 group, ◇ represents G2 group, ● represents G3 group, and ◆ represents G4 group.

[0029] Fig.12 The figure shows the area under the blood concentration-time curve (AUC) of vactosertib after oral administration of the test substance in the DSS-induced ulcerative colitis rat model. ○ represents the G1 group, ◇ represents the G2 group, ● represents the G3 group, and ◆ represents the G4 group.

[0030] Fig.13 The in vivo drug elimination half-life (t) of vactosertib after oral administration of the test substance in the DSS-induced ulcerative colitis rat model is shown. 1 / 2 ), ○ represents G1 group, ◇ represents G2 group, ● represents G3 group, and ◆ represents G4 group.

[0031] Fig.14 The following table shows the method for inducing ulcerative colitis and the administration schedule of the test substances used in animal experiments.

[0032] Fig.15 The severity of inflammation evaluated by histopathology in a mouse model of ulcerative colitis induced by DSS is shown. The data are shown as median values, 25-75% values ​​(in boxes), and minimum-maximum values ​​(two extremes), and the Dunn's test, a non-parametric test method built into Graphpad Prism 8.01, is used to test the statistical significance between the positive control group (▲G2 group) and the test substance administration group (◇G3 group, ●G4 group, ◆G5 group). The median values ​​of the inflammatory index of the bentonite administration group (G3 group) and the vactosertib administration group (G4 group) were slightly lower than those of the positive control group (G2 group), and the median value of the vactosertib / bentonite complex administration group (G5 group) was lower, but not statistically significant.

[0033] Fig.16 The severity of crypt cell damage assessed by histopathology in a mouse model of ulcerative colitis induced by DSS is shown. The data are shown as median values, 25-75% values ​​(in boxes), minimum-maximum values ​​(two extremes), and the Dunn's test method built into Graphpad Prism 8.01 version as a non-parametric test method is used to test the statistical significance between the positive control group (▲G2 group) and the test substance administration group (◇G3 group, ●G4 group, ◆G5 group). The median values ​​of the crypt cell damage index of the bentonite administration group (G3 group) and the vactosertib administration group (G4 group) were lower than those of the positive control group (G2 group), but there was no statistical significance, while the median value of the vactosertib / bentonite complex administration group (G5 group) was statistically significantly reduced.

[0034] Fig.17The severity of ulcers evaluated by histopathology in a mouse model of ulcerative colitis induced by DSS is shown. The data are shown as median values, 25-75% values ​​(in boxes), minimum-maximum values ​​(two extremes), and the Dunn's test method built into Graphpad Prism 8.01 version as a non-parametric test method is used to test the statistical significance between the positive control group (▲G2 group) and the test substance administration group (◇G3 group, ●G4 group, ◆G5 group). The median values ​​of the ulcer index in the bentonite administration group (G3 group) and the vactosertib administration group (G4 group) were lower than those in the positive control group (G2 group), but there was no statistical significance, while the median value of the vactosertib / bentonite complex administration group (G5 group) was statistically significantly reduced.

[0035] Fig.18 The severity of edema evaluated by histopathology in a mouse model of ulcerative colitis induced by DSS is shown. Data are shown as median values, 25-75% values ​​(in boxes), minimum-maximum values ​​(two extremes), and the Dunn test method built into Graphpad Prism 8.01 version as a non-parametric test method is used to test the statistical significance between the positive control group (▲G2 group) and the test substance administration group (◇G3 group, ●G4 group, ◆G5 group). Compared with the positive control group (G2 group), no significant changes can be observed in each test substance administration group.

[0036] Fig.19 The severity of fibrosis evaluated by histopathology in a mouse model of ulcerative colitis induced by DSS is shown. The data are shown as median values, 25-75% values ​​(in boxes), minimum-maximum values ​​(two extremes), and the Dunn's test method built into Graphpad Prism 8.01 version as a non-parametric test method is used to test the statistical significance between the positive control group (▲G2 group) and the test substance administration group (◇G3 group, ●G4 group, ◆G5 group). The median values ​​of the fibrosis index of the bentonite administration group (G3 group) and the vactosertib administration group (G4 group) were slightly lower than those of the positive control group (G2 group), and the median value of the vactosertib / bentonite complex administration group (G5 group) was lower, but there was no statistical significance.

[0037] Fig. 20A graph comparing the total severity scores of inflammation, crypt cell damage, ulcers and edema evaluated by histopathology in a mouse model of ulcerative colitis induced by DSS is shown (excluding fibrosis scores). Data are shown as median values, 25-75% values ​​(in boxes), minimum-maximum values ​​(two extremes), and the Dunn's test method built into Graphpad Prism 8.01 version as a non-parametric test method is used to test the statistical significance between the positive control group (▲G2 group) and the test substance administration group (◇G3 group, ●G4 group, ◆G5 group). The median values ​​of the ulcer index of the bentonite administration group (G3 group) and the vactosertib administration group (G4 group) were lower than those of the positive control group (G2 group), but there was no statistical significance, while the median value of the vactosertib / bentonite complex administration group (G5 group) was statistically significantly reduced.

[0038] Fig.21 The results of immunohistochemical staining are shown to show the effects of the test substances on interleukin-6 (IL-6) in the colon of the DSS-induced ulcerative colitis mouse model.

[0039] Fig. 22 The results are as follows: the IL-6 staining area was measured in the colon of a DSS-induced ulcerative colitis mouse model. DETAILED DESCRIPTION

[0040] The present invention relates to a controlled-release composition for oral administration, which comprises a complex of a compound of Chemical Formula 1 (hereinafter referred to as "vactosertib") or a pharmaceutically acceptable salt thereof and a clay mineral.

[0041] [Chemical formula 1]

[0042]

[0043] In addition, the present invention relates to a method for preparing a controlled-release composition for oral administration, comprising the steps of mixing a solution containing a compound of Chemical Formula 1 or a pharmaceutically acceptable salt thereof, an acidic reaction solution, and a clay mineral suspension to allow the compound to be adsorbed on the clay mineral to prepare a complex. The controlled-release composition for oral administration comprises a complex of the compound of Chemical Formula 1 and a clay mineral.

[0044] Furthermore, the present invention relates to a controlled release method, characterized in that a composition comprising a compound of Chemical Formula 1 or a pharmaceutically acceptable salt thereof and a clay mineral is orally administered to a subject.

[0045] [Chemical formula 1]

[0046]

[0047] Hereinafter, the present invention will be described in detail.

[0048] Compound of Chemical Formula 1

[0049] The compound of the chemical formula 1 of the present invention can be prepared by a known method. For example, the compound of the chemical formula 1 is N-((4-([1,2,4]triazolo[1,5-a]pyridin-6-yl)-5-(6-methylpyridin-2-yl)-1H-imidazol-2-yl)methyl)-2-fluoroaniline. The molecular formula of the above compound is C 22 H 18 FN7, the pH value of saturated aqueous solution at 25±3℃ is pH7.2, and the solubility (aqueous solubility) is <0.02mg / ml. In the low pH range, the solubility is higher (i.e., at pH2.81, the solubility is 25.05mg / ml), and at pH 4.9 and above, the solubility is less than 0.02mg / ml. Partition coefficient (octanol / water) Log D o / w The compound of Chemical Formula 1 can be prepared by the method disclosed in Korean Patent Publication No. 10-2013-0028749.

[0050] [Chemical formula 1]

[0051]

[0052] Clay Minerals

[0053] Generally, clay minerals have a layered structure of crystal units stacked by combining silicon sheets and aluminum sheets, i.e., a plate-like structure. In the clay minerals with interlayer expansion as described above, there is no hydrogen bond between the crystal units, and the binding force between the crystal units is weak, so when moisture is absorbed between these crystal units, expansion occurs. Therefore, even ions with larger sizes can easily enter between the crystal units of clay minerals with interlayer expansion. On the other hand, in clay minerals with interlayer expansion, tetrahedral Si with 4 valence positive charges is isomorphously replaced by Al or Fe with 3 valence positive charges, or octahedral Al or Fe3+ with 3 valence positive charges isomorphously replaced by Mg or Fe2+ with 2 valence positive charges to generate negative layer charges of the layers, but cations such as calcium ions (Ca2+), magnesium ions (Mg2+), sodium ions (Na+), and potassium ions (K+) are combined between the layers or on the surface, so the overall charge is neutral. The clay mineral of the present invention is excellent in adsorption of cationic substances, and the adsorption pattern changes depending on the surrounding pH value.

[0054] The clay mineral of the present invention has a plate-like structure, specifically, has interlayer expansion, and is a clay mineral that can insert antibiotics inside to serve as a carrier. The clay mineral of the present invention can be a smectite mineral, for example, it can be montmorillonite or bentonite, smectite, vermiculite, beidellite, nontronite, saponite, hectorite, etc. For example, the clay mineral of the present invention can be bentonite containing more than 50% by weight of montmorillonite. Preferably, the clay mineral of the present invention is bentonite.

[0055] Clay mineral / vactosertib complex

[0056] In the clay mineral / vactosertib complex of the present invention, clay mineral is used as a carrier that allows vactosertib to pass through the stomach (pH 1.5 to 2.0) and the small intestine (pH 7.2 to 7.5) and then pass to the colon (pH 7.9 to 8.5) to act in the colon. That is, the clay mineral / vactosertib complex of the present invention inhibits vactosertib from being absorbed in the upper digestive tract and delivers vactosertib to the colon. The clay mineral of the present invention is a layered clay mineral, the layered surface is negatively charged, and vactosertib is adsorbed on the clay mineral in a non-crystalline state. The above-mentioned complex can include clay minerals and compounds of chemical formula 1 or pharmaceutically acceptable salts thereof in a weight ratio of 1: 0.02 to 1: 0.26.

[0057] The clay mineral of the present invention is preferably bentonite. Under low pH conditions, weakly alkaline drug molecules with amine groups carry positive charges and can bind to bentonite, but under high pH conditions, they can lose their charges and be released from bentonite. Considering that the pH value of the upper digestive tract in the body is low and the pH value of the lower digestive tract is high, the bentonite / vactosertib complex of the present invention has pH-dependent release regulation capability.

[0058] That is, the dissolution rate of the complex of the present invention when placed in an eluent having a pH of 1.2 is slower than the dissolution rate of the compound powder of Chemical Formula 1 when placed in an eluent having a pH of 1.2 ( Figure 4 ). However, the dissolution rate of the above-mentioned complex when placed in a pH 7.4 dissolution solution is faster than the dissolution rate of the compound powder of Chemical Formula 1 when placed in a pH 7.4 dissolution solution ( Figure 6 ).

[0059] Although vactosertib is easily dissolved in an acidic environment, it is neutrally charged at pH 7, and its solubility in neutral is low, so its absorption is concentrated in the upper digestive tract, and it has an absorption problem that it cannot be fully absorbed in the lower digestive tract, which is the target organ. That is, when vactosertib is orally administered, it is positively charged by gastric acid to increase its solubility, and then when passing through the small intestine, the pH value returns to neutral again, so that some neutral molecules are absorbed in the upper part of the small intestine. Therefore, in terms of pharmacokinetics, there is a problem of a sharp increase in the initial blood drug concentration of vactosertib after oral administration. In addition, after vactosertib is neutralized in the small intestine, its solubility decreases sharply and precipitates, and it cannot be absorbed in the small intestine afterwards. As a result, the blood drug concentration decreases sharply in terms of pharmacokinetics. That is, the charge and solubility of vactosertib change with the change of pH value, resulting in a disadvantage that the area where it can be absorbed in the small intestine is very limited. Furthermore, the dissolution and precipitation of vactosertib are greatly affected by gastric and intestinal juices, and since the degree of secretion of gastric and intestinal juices varies from person to person, it is difficult to expect a prescribed effect in patients. In addition, vactosertib is taken in the form of a non-crystalline solid dosage form with aggregated molecules, so it requires a process of dissolving it into individual molecules by gastric acid. Furthermore, since the half-life of the drug itself is very short, even if the drug is quickly absorbed in the upper gastrointestinal tract, it is difficult to expect a long-term effect of the pharmacological effect.

[0060] However, in the case of a clay mineral / vactosertib complex, preferably, in the case of a bentonite / vactosertib complex in which vactosertib is adsorbed on bentonite, vactosertib is inserted into the interlayer of bentonite as a cation-state molecule, and there is no process of dissolving in gastric acid. In addition, bentonite has a coating ability, and after being adsorbed on the epithelial cells of the small intestine, vactosertib as a drug is separated and neutralized and then absorbed, so it can be absorbed by the entire area of ​​the small intestine. In addition, the clay mineral / vactosertib complex that moves to the colon adsorbs and releases vactosertib directly to the affected area, thereby having the effect of delivering the drug to the affected area in a dual manner.

[0061] Compositions comprising a clay mineral / vactosertib complex

[0062] The present invention relates to a controlled-release composition for oral administration, which comprises a complex of a compound of Chemical Formula 1 or a pharmaceutically acceptable salt thereof and a clay mineral.

[0063] Vactosertib is easily dissolved in an acidic environment, but has low solubility in a neutral environment, so it has a problem of uncertain absorption in the body. That is, in the case of the upper digestive tract including the stomach, the pH is very low at about 1 to 3, and when passing through the small intestine from the duodenum, the pH increases to a level of 5 to 8. Therefore, vactosertib is absorbed only in the upper digestive tract, and it is often not fully absorbed in the lower digestive tract. In addition, the half-life of the drug itself is very short, so even if the drug is quickly absorbed in the upper digestive tract, it is difficult to expect a long-term effect of the pharmacological effect. In contrast, in the case of a composition containing a clay mineral / vactosertib complex, vactosertib is ionized in an acidic pH environment, so it is adsorbed to the clay mineral at a low pH and released at a high pH. Therefore, vactosertib is stably delivered to the lower digestive tract with a high pH value, such as the colon, in a state adsorbed to the clay mineral, so it not only delays the release of the drug, but also helps vactosertib to continue to work in the colon.

[0064] Furthermore, under acidic conditions, the clay mineral / vactosertib complex of the present invention maintains strong adsorption between vactosertib and clay mineral, thereby preventing rapid absorption of the drug after administration. Furthermore, the clay mineral / vactosertib complex can be well dispersed in water, so it is easier to absorb than tablets and can be implemented in a reproducible manner.

[0065] Furthermore, the clay mineral / vactosertib complex of the present invention is different from the conventional enteric polymers such as cellulose acetate phthalate, hydroxypropylmethylcellulose phthalate acetate, polyvinyl acetate phthalate, polyacrylate L and polyacrylate S series polymers, and does not require a coating step for preventing the initial release of the drug.

[0066] The peak time (t max ) can be 30 minutes to 60 minutes. The peak time of blood drug concentration when orally administered to humans (t max ) can be 3 hours to 10 hours. In addition, the peak time of blood drug concentration (t max ) is longer than the time to peak blood concentration (t max Preferably, if the dosage of the compound of Chemical Formula 1 is the same, the peak time of blood concentration (t max ) is shorter than the time to peak blood concentration (t max) is 2 to 10 times longer, more preferably, 2.3 to 8 times longer, even more preferably, 2.4 to 6 times longer, further preferably, 2.6 to 5 times longer, and further more preferably, 2.7 to 4 times longer.

[0067] The peak drug concentration (C max ) may be 0.7 μg / ml to 1.9 μg / ml. Furthermore, the peak drug concentration (C max ) is lower than the peak drug concentration (C max Preferably, if the dosage of the compound of Chemical Formula 1 is the same, the peak drug concentration (C max ) is the peak drug concentration (C max ), more preferably 20% to 80% of the total weight of the raw material, more preferably 25% to 70%, even more preferably 30% to 60%, further preferably 35% to 50%, and further more preferably 37% to 45%.

[0068] The dissolution rate of the complex of the present invention when placed in a pH 1.2 dissolution solution is slower than the dissolution rate of the compound powder of Chemical Formula 1 when placed in a pH 1.2 dissolution solution. However, the dissolution rate of the above-mentioned complex when placed in a pH 7.4 dissolution solution is faster than the dissolution rate of the compound powder of Chemical Formula 1 when placed in a pH 7.4 dissolution solution. In addition, the absorption rate of the compound of Chemical Formula 1 in the small intestine when the above-mentioned composition is orally administered is lower than the absorption rate in the small intestine when an aqueous solution of the compound of Chemical Formula 1 is orally administered. The above-mentioned absorption rate refers to the amount of the compound absorbed relative to the amount of the compound of Chemical Formula 1 administered per unit time. The reduced absorption amount of the compound of Chemical Formula 1 in the small intestine when the above-mentioned composition is orally administered means that the amount of the compound of Chemical Formula 1 that acts in the colon can be relatively increased. In addition, the absorption rate (rate) of the compound of Chemical Formula 1 in the small intestine when the composition of the present invention is orally administered is slower than the absorption rate in the small intestine when an aqueous solution of the compound of Chemical Formula 1 is orally administered. In this case, the change in the absorption rate in the small intestine can be measured using the peak time of blood drug concentration (t max ) and peak drug concentration (C max For example, if the peak time of blood drug concentration (t max ) increases, peak drug concentration (C max ) decreases, which means that the absorption rate decreases. If the peak time of blood drug concentration (t max ) is reduced, and the peak concentration (C max) increases, which means that the absorption rate increases. Therefore, the composition of the present invention is suitable for releasing the compound of Chemical Formula 1 in the colon. When the composition of the present invention is orally administered, the drug improvement effect of the compound of Chemical Formula 1 in the colon on the symptoms selected from the group consisting of inflammation, crypt cell damage, ulcers, edema and fibrosis is better than the drug improvement effect of the compound of Chemical Formula 1 in the colon on the symptoms selected from the group consisting of inflammation, crypt cell damage, ulcers, edema and fibrosis when the aqueous solution of the compound of Chemical Formula 1 is orally administered. Preferably, when the composition of the present invention is orally administered, the drug improvement effect shown in the total score of inflammation, crypt cell damage, ulcers, edema and fibrosis in the colon recorded in Table 8 is better than the drug improvement effect shown in the total score of inflammation, crypt cell damage, ulcers, edema and fibrosis in the colon recorded in Table 8 when the aqueous solution of the compound of Chemical Formula 1 is orally administered. In particular, the composition of the present invention can deliver a drug composition with the colon as the target organ to the colon to increase the drug effect in the colon.

[0069] The composition of the present invention is a controlled-release composition. Controlled-release (modified-or controlled-release) means that after administration of the drug, the blood concentration of the drug rises rapidly to an effective concentration and only maintains a prescribed blood concentration of the drug for a desired period of time. Compared with other common preparations, the administration frequency is low, the in vivo reaction is uniform, and the side effects are few.

[0070] The composition of the present invention may be a pharmaceutical composition. The pharmaceutical composition of the present invention may contain 0.01 weight percent to 80 weight percent of a compound of Formula 1 or a pharmaceutically acceptable salt thereof and a complex of a clay mineral, preferably 0.02 weight percent to 65 weight percent. However, the above content may be increased or decreased according to the needs of the person administering the drug, and may be appropriately increased or decreased according to conditions such as age, diet, nutritional status, and the course of the disease. A person of ordinary skill in the art to which the present invention belongs may appropriately determine the content of the compound of Formula 1 or a pharmaceutically acceptable salt thereof and a complex of a clay mineral contained in the pharmaceutical composition of the present invention.

[0071] The pharmaceutical composition of the present invention can be administered orally and can be used as a common pharmaceutical preparation. For example, the pharmaceutical composition of the present invention can be used in the form of a preparation for oral administration such as tablets, granules, capsules, suspensions, etc. The above preparation can be prepared using an acceptable common carrier. For example, in the case of a preparation for oral administration, an excipient, a binder, a disintegrant, a lubricant, a dissolving agent, a coloring agent, a coating agent, a suspending agent, a preservative or a bulking agent can be used to prepare it. The above is only an example of the preparation form and additives that can be used to prepare the pharmaceutical composition of the present invention, and the pharmaceutical composition of the present invention is not limited thereto.

[0072] The dosage of the pharmaceutical composition of the present invention can be determined by experts according to various factors such as the patient's condition, age, gender and complications, but can usually be administered at a dosage of 0.1 mg to 10 g per kilogram of body weight for adults, preferably, 10 mg to 5 g per kilogram of body weight. In addition, the unit dosage form can contain a daily dose of the above-mentioned pharmaceutical composition or 1 / 2, 1 / 3 or 1 / 4 of the dose thereof, and can be administered once to 6 times a day, but is not limited thereto, and the attending physician can appropriately adjust the above dosage.

[0073] Preparation method of the composition of the present invention

[0074] The present invention relates to a method for preparing a controlled release composition for oral administration, which comprises mixing a solution containing a compound of Chemical Formula 1 or a pharmaceutically acceptable salt thereof, an acidic reaction solution and a clay mineral suspension and adsorbing the compound on the clay mineral to prepare a complex, wherein the controlled release composition for oral administration comprises a complex of the compound of Chemical Formula 1 and the clay mineral. In this case, in order to adsorb the compound of Chemical Formula 1 on a layered clay mineral in a non-crystalline state, a method of mixing the compound and the clay mineral by dissolving and dispersing them in the same aqueous solution can be used. In this case, since the layered surface of the clay mineral (e.g., bentonite) carries a negative charge, if the drug dissolved in the aqueous solution carries a cation, it can be more effectively adsorbed.

[0075] Because the solubility of vactosertib in a neutral pH environment is very low, it is difficult to be ionized under the corresponding pH conditions to be adsorbed on clay minerals. However, when the pH value of the reaction solvent is acidic, the solubility of vactosertib increases, and a large amount of drugs can be reacted with bentonite. Due to the structural characteristics of vactosertib, it is positively charged at low pH values ​​and is bound to clay minerals with a high adsorption rate through a stronger ion binding reaction. Therefore, preferably, the pH value of the above-mentioned reaction solution is pH 0.5 to pH 3.5. In this case, in order to impart acidic conditions to the reaction solution, one of hydrochloric acid, phosphoric acid, sulfuric acid, acetic acid, and formic acid can be selected. Moreover, the solubility of vactosertib at a pH of pH 3 is about 1 mg / mL, and considering the situation that the lower the pH value, the more the solubility increases, it is more preferred that the reaction pH value is selected in the range of 1 to 3.

[0076] In this case, for more sufficient adsorption, preferably, the above-mentioned complex contains the compound of Chemical Formula 1 or a pharmaceutically acceptable salt thereof and a clay mineral in a weight ratio of 0.1:5 to 1.3:5. If an excessive amount of the drug (the compound of Chemical Formula 1) reacts in the clay mineral, the amount of drug lost without being attached to the clay mineral increases. On the contrary, if the amount of clay mineral is higher than the amount of drug, although the overall adsorption rate can be improved, the drug content in the final obtained complex is low and the dosage is increased, which will bring a burden to the administration.

[0077] In order to obtain a complex in a dry form after forming the complex of the present invention under specified conditions, a variety of methods can be used. In addition, after vactosertib is adsorbed on a clay mineral, unbound vactosertib can be removed by centrifugation, removal of the supernatant, and the like. In addition, when the precipitate is freeze-dried, a powder-shaped complex with properties similar to those of existing clay minerals can be obtained. On the other hand, in order to separate the reaction solution from the complex, a process of passing only the solvent through a filter and repeatedly washing the complex precipitate that has not passed through with distilled water can be included, and the unadsorbed drug and acidic solution can be removed without centrifugation through the above process. Alternatively, a method of relieving the strong acidic conditions of the solution and freeze drying after the adsorption reaction is completed can also be used. Specifically, after reacting at a low pH value that enables effective adsorption of the drug, freeze drying is performed after increasing the pH value of the reaction solution, thereby reducing the possibility of corrosion of the instrument and allowing the freeze dryer to run smoothly.

[0078] The composition of the present invention can be applied to various diseases for which vactosertib is known to have preventive, ameliorative or therapeutic effects. For example, as described in Korean Patent No. 10-1500665, the composition of the present invention is effective for various disease states mediated by anaplastic lymphoma kinase 5 (ALK5) and / or anaplastic lymphoma kinase 4 (ALK4), such as glomerulonephritis, diabetic retinopathy, lupus nephritis, hypertension-induced renal disease, renal interstitial fibrosis, renal fibrosis caused by drug-induced complications, renal disease associated with immunodeficiency syndrome (HIV), transplanted renal disease, liver fibrosis caused by any disease, liver dysfunction caused by hepatitis, alcoholic hepatitis, biliary disease, cystic fibrosis, pulmonary fibrosis, interstitial lung disease, acute lung injury, adult respiratory distress syndrome, idiopathic pulmonary fibrosis, chronic obstructive pulmonary disease, lung disease caused by infectious agents or toxic agents, cardiac fibrosis after myocardial infarction, congestive heart failure, dilated cardiomyopathy, myocarditis, vascular intimal thickening, vascular stenosis, hypertension-induced angiogenesis, pulmonary hypertension, coronary restenosis, peripheral vascular remodeling, etc. Stenosis, carotid restenosis, stent-induced restenosis, atherosclerosis, eye scars, corneal scars, proliferative vitreoretinopathy, glaucoma, intraocular pressure, excessive or hypertrophic scars or keloids formed in the skin during the healing process after trauma or surgery, membrane and subcutaneous stenosis, scleroderma, fibrosclerosis, progressive systemic sclerosis, dermatomyositis, polymyositis, arthritis, osteoporosis, ulcers, nerve damage, male impotence, Peyronie's disease The present invention can prevent, treat or improve the following diseases: Alzheimer's disease, Dupuytren's contracture, Alzheimer's disease, Raynaud's syndrome, radiation-induced fibrosis, thrombosis, tumor metastatic growth, multiple myeloma, melanoma, glioma, glioblastoma, leukemia, sarcoma, leiomyoma, mesothelioma, and tumors of the lung, breast, colon, kidney, ovary, neck, liver, bile duct, gastrointestinal tract, pancreas, prostate, head and neck. At the same time, it has a preventive, therapeutic or improving effect on myelodysplastic syndrome or lymphoma. In addition, the composition of the present invention can be a pharmaceutical composition with the following digestive tract, such as colon as the target organ. For example, the composition of the present invention can be a composition for preventing, improving or treating inflammatory bowel disease. For example, the composition of the present invention can be a composition for preventing, improving or treating Crohn's disease or ulcerative colitis.

[0079] <Materials and Methods>

[0080] The compound of Chemical Formula 1 (hereinafter referred to as "vactosertib") was obtained from MedPacto Co., Ltd., Korea. Bentonite was obtained from the Korea Institute of Geosciences and Mineral Resources and used.

[0081] <Experimental Example 1>

[0082] <1-1>

[0083] The adsorption rate of vactosertib and bentonite according to the pH value of the solvent was evaluated. In this case, considering that vactosertib is not easily soluble in a neutral environment, but starts to dissolve at a solubility of about 1 mg / mL under the condition of pH 3, the reaction pH values ​​were selected as pH 1, pH 2, and pH 3 for testing.

[0084] First, vactosertib was dissolved in a 0.1N hydrochloric acid aqueous solution at a concentration of 10 mg / mL, and diluted in a hydrochloric acid aqueous solution or a phosphate buffer corresponding to each pH value (pH 1: 0.1N hydrochloric acid aqueous solution, pH 2, pH 3: phosphate buffer) to prepare a concentration of 1 mg / mL. 1 mL of the relevant drug solution, 8 mL of the reaction solution corresponding to each pH value (i.e., 0.1N hydrochloric acid aqueous solution or phosphate buffer) and 1 mL of a suspension prepared by suspending bentonite in distilled water to 5 mg / mL were mixed and left for 30 minutes to allow the drug to be adsorbed to the bentonite. Then, the drug concentration of the supernatant obtained by centrifugation (3,000 rpm, 5 minutes) was analyzed. In this way, the amount of drug remaining in the supernatant was subtracted from the total drug used (vactosertib), and the amount of drug adsorbed to the bentonite was indirectly calculated by mathematical formula 1 to calculate the adsorption rate.

[0085] [Mathematical formula 1]

[0086]

[0087] As a result, although the difference between the groups was not significant, it was confirmed that the adsorption rate slightly increased with the decrease in pH value, and the adsorption efficiency of vactosertib was the highest at pH 1 (Table 1). Therefore, in the following experiment, when preparing the complex of vactosertib and bentonite, the pH value of the solvent was set to pH 1 for the experiment.

[0088] [Table 1]

[0089]

[0090] The above concentrations are based on the final concentrations.

[0091] <1-2>

[0092] The adsorption rate according to the ratio of vactosertib to bentonite was evaluated in order to find the composition that maximizes the adsorption to bentonite while minimizing the loss of vactosertib.

[0093] To this end, the sample was prepared and prepared in the same manner as in Experimental Example 1, the reaction concentration of bentonite and the reaction pH were fixed at 0.5 mg / mL and pH 1, respectively, and the concentration of vactosertib used was prepared at different concentrations from 0.01 mg / mL to 0.5 mg / mL to prepare the composite. Then, the difference in adsorption rate according to the concentration of vactosertib was detected.

[0094] As a result, when the concentration of vactosertib was low, although most of the drug remained bound to the bentonite, the adsorption rate began to decrease from a vactosertib concentration of 0.1 mg / mL, and at concentrations above this concentration, the drug adsorption rate was 60% or less, which was very low (Table 2). Therefore, the composition that ensured a sufficiently high adsorption rate and a high content of vactosertib in the complex, that is, a ratio of vactosertib concentration of 0.025 mg / ml to 0.1 mg / ml relative to a bentonite concentration of 0.5 mg / ml was selected as the optimal bentonite / vactosertib ratio.

[0095] [Table 2]

[0096]

[0097] <1-3>

[0098] Under the conditions where the reaction pH and the ratio of bentonite to vactosertib were fixed, the adsorption rate was evaluated according to the absolute concentration changes of vactosertib and bentonite used.

[0099] To this end, the sample was prepared in the same manner as in Experimental Example 1, and the concentration ratio of vactosertib and bentonite was adjusted to 1:5 using a solvent at pH 1, and the absolute concentration (ie, actual concentration) was adjusted to be different to prepare the composite. The difference in adsorption rate according to the change in absolute concentration was detected.

[0100] As a result, as the absolute concentrations of vactosertib and bentonite used in the complex increased by 10 times, the drug adsorption rate increased, thereby confirming that the loss of the drug could be reduced (Table 3).

[0101] [Table 3]

[0102]

[0103] <Experimental Example 2>

[0104] <2-1>

[0105] The bentonite / vactosertib complex was prepared using the method of Experimental Example 1 using a solvent at pH 1, a vactosertib concentration of 1 mg / ml and a bentonite concentration of 5 mg / ml. Then, after freeze-drying, vactosertib was extracted and analyzed to detect the actual content of vactosertib.

[0106] The freeze-dried complex was dispersed in phosphate buffered saline (pH 7.4) containing 0.5% Tween 80 to extract vactosertib. The calculated reaction weight percentage of the bentonite / vactosertib complex was approximately 16.9±0.2%, which is similar to the reaction weight percentage of existing vactosertib and bentonite. The reaction weight percentage was calculated by the following mathematical formula 2.

[0107] [Mathematical formula 2]

[0108]

[0109] <2-2>

[0110] The morphological properties of the bentonite / vactosertib complex prepared in <2-1> were evaluated. To this end, the bentonite / vactosertib complex prepared in <2-1> and vactosertib were attached to copper tapes, respectively, and the surfaces were coated with platinum, and then photographed using a scanning electron microscopy (SEM).

[0111] As a result, it was observed that the vactosertib powder had a unique crystal shape with a size of 1 μm or more ( Figure 1 ). However, the bentonite / vactosertib complex did not show such a crystalline shape, but rather showed a morphology in which the layered structure of bentonite was irregularly aggregated together ( Figure 2 ).

[0112] <2-3>

[0113] The crystallographic properties of the bentonite / vactosertib complex were analyzed by XRD. The XRD patterns of vactosertib, bentonite, a physical mixture of vactosertib and bentonite, and the bentonite / vactosertib complex were analyzed and compared to confirm the changes in the crystal structure according to the drug adsorption phenomenon.

[0114] As a result, in the case of vactosertib, there is a unique XRD curve, which also shows similar behavior in the physical mixture. However, in the case of bentonite / vactosertib composite, no unique peak is shown, and a pattern very similar to that of existing bentonite is confirmed ( Figure 3 ).

[0115] Figure 3 The XRD patterns of each substance are shown in a graph. In the case of vactosertib, there is a unique XRD peak pattern, which also has similar performance in physical mixtures. However, in the case of bentonite / vactosertib composite, no unique peak is shown, and it is confirmed that the pattern is very similar to the pattern of existing bentonite.

[0116] <Experimental Example 3>

[0117] <3-1>

[0118] In order to confirm the changes in the release of vactosertib after adsorption to bentonite, a dissolution test of vactosertib was performed.

[0119] In this case, in order to predict the drug release in the upper and lower digestive tracts, dissolution was performed under different pH conditions. For this purpose, 0.1N HCl solution (pH 1.2), 50mM acetate buffer (pH 4.5), and 50mM phosphate buffer (pH 7.4) were used as dissolution solutions for testing. In this case, in order to provide a dissolution environment for sufficient dissolution of vactosertib, Tween 80 was dissolved in all dissolution solutions at a concentration of 0.1% and used.

[0120] Vactosertib powder and bentonite / vactosertib complex were prepared as dispersions in a manner that the concentration of vactosertib was 1 mg / mL, and 0.3 mL of the dispersions were sealed in a semipermeable membrane and immersed in 30 mL of each dissolution solution for dissolution. Then, 0.2 mL of the dissolution solution was collected at different specified times, and the dissolution rate was compared by analyzing the drug concentration.

[0121] As a result, the vactosertib powder dissolved rapidly at pH 1.2, but the dissolution rate slowed down significantly at higher pH values. In contrast, the bentonite / vactosertib complex showed a lower dissolution rate than the existing vactosertib powder at both pH 1.2 and pH 4.5, but a higher dissolution rate at pH 7.4 ( Figure 4 : pH 1.2, Figure 5 : pH 4.5, Figure 6 : pH 7.4). Therefore, it was confirmed that when vactosertib, which can be rapidly dissolved in the upper gastrointestinal tract, is adsorbed on bentonite, the dissolution and absorption of vactosertib can be reduced, and vactosertib can be dissolved and absorbed in the lower gastrointestinal tract having a higher pH value.

[0122] <3-2>

[0123] In order to confirm the change of blood drug concentration during the actual oral administration of bentonite / vactosertib complex, a drug dynamic analysis test was performed using SD rats of about 7 weeks of age. The existing vactosertib powder and the aqueous solution dissolved in the pH 3 buffer solution, as well as the bentonite / vactosertib complex were used as samples for testing, and the dose was set to 10 mg / kg based on the amount of the drug (i.e., vactosertib). After oral administration of each sample to rats, blood was collected at different time points, and pre-treatment was performed to analyze blood drug concentration.

[0124] As a result, compared with the existing vactosertib powder, when used in the form of an aqueous solution, the absorption of vactosertib as a drug became higher, reaching the peak drug concentration in 15 minutes. On the other hand, in the case of the bentonite / vactosertib complex in which vactosertib was adsorbed on bentonite, not only was the peak drug concentration significantly reduced compared to the vactosertib aqueous solution, but the time to reach the peak drug concentration was slower, and the blood drug concentration in the latter half was higher than that of the aqueous solution. Moreover, overall, the area under the blood drug concentration curve (AUC) of the bentonite / vactosertib complex group was larger than that of the vactosertib powder group. This confirms that the bentonite / vactosertib complex exhibits better absorption than the existing vactosertib powder, and can slow down the absorption of the drug (i.e., vactosertib) compared to the rapidly absorbed aqueous solution form of vactosertib ( Figure 7 ).

[0125] Based on the above Figure 7The results of the in vivo dynamics of each group were analyzed. The results showed that the area under the blood concentration curve of the group using vactosertib in powder form was significantly lower than that of the other groups, and the deviation value compared with the average value of each parameter was higher than that of the other groups. When vactosertib was used in the form of an aqueous solution, the highest C max (peak drug concentration) and the fastest T max (Time to Peak Blood Drug Concentration), which indicates that the drug is absorbed quickly in the solution state. On the other hand, in the case of bentonite / vactosertib complex, compared with vactosertib aqueous solution, although C max Lower, but T max That is, when the bentonite / vactosertib complex is orally administered, the time to peak blood drug concentration (T max ) was 45±18.37 minutes, which was shorter than the peak time of plasma concentration (T max )15±0 minutes is about 3 times longer. In addition, when the bentonite / vactosertib complex is orally administered, the peak drug concentration (C max ) was 1.12±0.37μg / mL, which was the peak concentration (C max )2.77±0.55μg / mL is about 40.4%. It was confirmed that the absorption of vactosertib can be delayed by adsorbing vactosertib to bentonite. Compared with vactosertib powder, the deviation value of the drug dynamic blood pressure parameter of the bentonite / vactosertib complex was also low, and it was known that the bentonite / vactosertib complex was more uniformly absorbed than vactosertib (Table 4).

[0126] [Table 4]

[0127]

[0128] <Experimental Example 4>

[0129] Vactosertib was dissolved in 0.1N aqueous hydrochloric acid solution at a concentration of 10 mg / mL, and diluted to a concentration of 1 mg / mL using 0.1N aqueous hydrochloric acid solution at pH 1. 1 mL of the corresponding drug (i.e., vactosertib) solution, 8 mL of 0.1N aqueous hydrochloric acid solution, and 1 mL of a suspension prepared by suspending bentonite in distilled water to 5 mg / mL were mixed and allowed to stand for 30 minutes to allow the drug to be adsorbed on the bentonite. Then, centrifugation (3,000 rpm, 5 minutes) was performed to obtain a supernatant, which was freeze-dried to prepare a bentonite / vactosertib complex.

[0130] <Experimental Example 5>

[0131] <5-1>

[0132] The biopharmaceutical differences of vactosertib dosage forms combined with or without bentonite were evaluated by studying the pharmacodynamics of vactosertib in a rat model of ulcerative colitis induced by DSS (dextran sulfate sodium).

[0133] First, dissolve vactosertib in 0.1N hydrochloric acid aqueous solution to a concentration of 10 mg / mL, and then dilute it with pH 3.0 phosphate buffer to prepare a solution with a concentration of 1 mg / mL for use. Suspend the vactosertib-bentonite complex in distilled water at 5.92 mg / mL to make it the same drug concentration as the vactosertib aqueous solution. If all preparations are ultrasonically treated with a Bathsonicator, they will be better dispersed and easily precipitated after dispersion, so they should be shaken thoroughly before administration.

[0134] The adsorption amount and drug release of the prepared bentonite / vactosertib complex were analyzed by HPLC in vitro. The analysis of in vivo samples such as plasma analysis, which is equivalent to low-concentration analysis, was performed by liquid chromatography-mass spectrometry (LC / MS / MS). Each plasma sample was mixed with acetonitrile in a certain ratio to remove protein, and only the supernatant was taken for analysis after centrifugation.

[0135] SD rats were used as test animals. The SD rats were obtained from the Gapyeong Center of Orient Bio Co., Ltd. (699-13, Mokdong-ri, Buk-myeon, Gapyeong-gun, Gyeonggi-do, South Korea) and were about 6 weeks old when purchased. After about 3 days of quarantine and acclimation after purchase, only healthy animals were used for the experiment. When the test substance was administered, the rats were about 8 weeks old, and 40 rats were used. The test animals were raised in a clean animal room on the second floor of the Life Science Laboratory of the Institute of Medicine, Inha University, and were ventilated at least 10 times / hour, at a temperature of 22±4°C, a relative humidity of 50±20%, and at a lighting of 12 hours (lighting: 08:00-20:00) and an illumination of 150Lux-300Lux. Solid feed irradiated with rodents (PMI) was used. 5053, USA), the experimental animals were allowed to drink freely using a feeder, and the tap water was treated with reverse osmosis, sterilized using an ultraviolet water sterilizer and high-pressure steam, and then provided to the experimental animals freely using a polysulfone drinking bottle (500 ml).

[0136] To induce ulcerative colitis, 5.5% sodium dextran sulfate was added to the drinking water of rats and continuously supplied for 6 days. The test substance was orally administered once after an overnight (12 hours) fast. The blood was also fasted during the blood collection. The test groups are shown in Table 5 below. In this case, G1 was supplied with a normal diet from the first day (DAY 1) to the sixth day (DAY 6), and vactosertib was orally administered once on the seventh day (DAY 7) after an overnight fast. G2 was supplied with a normal diet from the first day to the sixth day, and a single oral administration of bentonite / vactosertib complex was administered on the seventh day after an overnight fast. G3 was induced from the first day to the sixth day with 5.5% sodium dextran sulfate, and a single oral administration of vactosertib was administered on the seventh day after an overnight fast. G4 was induced from the first day to the sixth day with 5.5% sodium dextran sulfate, and a single oral administration of bentonite / vactosertib complex was administered on the seventh day after an overnight fast. The above administration was performed by placing the test substance into a syringe and then connecting it to a rat feeding tube for oral administration. Then, vactosertib or bentonite / vactosertib complex was administered once according to the group ( Figure 8 ).

[0137] [Table 5]

[0138]

[0139] The number of animals that can be finally evaluated is shown in ().

[0140] Clinical symptom observation is to observe general symptoms (whether there is blood in stool, diarrhea, etc.) and whether there is death / dying once a day for different individuals during the test. In this case, only individuals with specific symptoms are kept records (normal cases are not recorded). In addition, the body weight of the test animals is measured every day before purchase, grouping and administration of test substances.

[0141] The results of the purchase of test animals, grouping, and measurement of the average body weight of each group before administration of the test substance showed no statistically significant difference between the groups. However, the average body weight of the G3 and G4 groups (dextran sulfate sodium induced groups) measured on the day of autopsy (seventh day) showed a statistically significant decrease compared with the G1 and G2 groups (distilled water induced groups).

[0142] (p<0.05, Table 6).

[0143] [Table 6]

[0144]

[0145] Body weight during the test

[0146] The data were expressed as mean ± standard deviation, and statistical analysis was performed using SPSS statistic 19 (Mann-Whitney test).

[0147] *p<0.05, significantly different from group G1

[0148] # p<0.05, significantly different from group G2

[0149] Group G1: n=10, group G2: n=9, group G3: n=9, group G4: n=10.

[0150] <5-2>

[0151] The dynamics of the drug in vivo were evaluated by detecting the changes in the blood concentration of vactosertib over time. Fig. 9 As shown in the figure (○ represents G1 group, ◇ represents G2 group, ● represents G3 group, ◆ represents G4 group), the area under the curve of blood drug concentration (AUC, area under the concentration-time curve) as an indicator of the absorption amount of the drug component in the body, the time to peak blood drug concentration (t max ) and peak drug concentration (C max , maximum plasma concentration).

[0152] The results showed that the peak time of blood drug concentration in G4 group was t max Increase, peak drug concentration C max In addition, it was confirmed that the peak drug concentration C in the G4 group was higher than that in the G3 group. max After that, the second half of the time-concentration curve maintains a higher blood concentration of the drug (i.e., vactosertib) for a long time ( Fig. 9 ).

[0153] Specifically, the peak time of blood drug concentration (t max ), the time to peak plasma drug concentration was prolonged by approximately 2- to 4-fold in most animals given the bentonite / vactosertib complex ( Fig.10 , ○ represents G1 group, ◇ represents G2 group, ● represents G3 group, ◆ represents G4 group). This means that the drug is slowly released in the gastrointestinal tract to prolong the absorption time of the drug, thereby increasing the duration of the drug effect.

[0154] On the other hand, at the peak drug concentration (C max), the peak drug concentration was confirmed to be reduced to less than half in the group administered with bentonite / vactosertib complex ( Fig.11 , ○ represents G1 group, ◇ represents G2 group, ● represents G3 group, ◆ represents G4 group). Therefore, by combining vactosertib with bentonite, a sharp increase in blood drug concentration can be prevented and the risk of adverse reactions caused by the administered drug can be reduced. In addition, in the bentonite / vactosertib complex administration group, the peak blood drug concentration time (t max ) becomes longer, and the blood drug concentration decreases slowly, confirming the advantage of reducing the frequency of drug administration.

[0155] In the case of the area under the plasma concentration-time curve (AUC), the combination of vactosertib and bentonite did not significantly affect the area under the plasma concentration-time curve of vactosertib ( Fig.12 , ○ represents G1 group, ◇ represents G2 group, ● represents G3 group, ◆ represents G4 group). Therefore, the combination with bentonite maintains the total drug exposure in the blood and prolongs the duration of drug effect, and it was confirmed that even if vactosertib is combined with bentonite, the amount of vactosertib entering the body does not show a significant difference.

[0156] The elimination half-life of the drug in the body (t 1 / 2 ), the combination of vactosertib and bentonite did not significantly affect the half-life ( Fig.13 , ○ represents G1 group, ◇ represents G2 group, ● represents G3 group, ◆ represents G4 group). Therefore, it was confirmed that this was not related to the slow absorption of the drug, and a similar rate of drug disappearance in the body could be maintained.

[0157] <Experimental Example 6>

[0158] Animal experiments were conducted using a mouse model of ulcerative colitis, one of the inflammatory bowel diseases, to test the anti-inflammatory, anti-crypt cell damage, anti-ulcer, anti-edema, and anti-fibrosis effects of the vactosertib / bentonite complex. The inflammation induction method and drug administration regimen used in the animal experiments were as follows: Fig.14As shown. C57BL / 6 mice were selected as experimental animals. In order to induce ulcerative colitis, the following cycle was repeated: sodium dextran sulfate, an inflammatory inducing substance, was added to the food and supplied for 7 days, and then a normal diet was supplied for 14 days. After repeating this inflammatory induction cycle three times, chronic inflammation, ulcers and fibrosis in the colon were induced. Starting from 3 weeks after the end of the first inflammatory induction cycle, bentonite, vactosertib or vactosertib / bentonite complex was administered to the experimental animal group other than the control group twice a day (10 am and 4 pm) and 5 days a week for a total of 6 weeks. The converted dose was based on per kilogram of mouse body weight, with bentonite being 106.8 mg, vactosertib being 20.0 mg, and the complex being 126.8 mg. In this case, the complex prepared in Experimental Example 5 was used as a vactosertib / bentonite complex. The inflammatory inducing substances, test substances, test substance doses and the number of test animals in the different test groups used in the animal test of ulcerative colitis are shown in Table 7 below.

[0159] [Table 7]

[0160]

[0161] The number of animals that can be finally evaluated is shown in ().

[0162] The C57BL / 6 mice used in the experiment were obtained from the Gapyeong Center of Orient Bio (699-13, Mudong-ri, Buk-myeon, Gapyeong-gun, Gyeonggi-do, South Korea) of Korea Co., Ltd. and were about 6 weeks old when purchased. After about 7 days of quarantine and acclimatization after purchase, only healthy animals were used for the experiment. The experimental animals were about 8 weeks old when the inducer was administered, and 52 were used. The experimental animals were raised in a clean animal room on the second floor of the Life Science Laboratory of the Institute of Medicine, Inha University, and were ventilated at least 10 times / hour, at a temperature of 22±4°C, a relative humidity of 50±20%, and 12 hours of lighting (lighting: 08:00-20:00) and an illumination of 150Lux-300Lux. The experimental animals were allowed to freely consume the irradiated solid feed for rodents (FeedLabGMO Free Rodent Diet GF2005) using a feeder, and the tap water was subjected to reverse osmosis treatment, sterilized using an ultraviolet water sterilizer and high-pressure steam, and then freely consumed by the experimental animals using a polysulfone drinking bottle (300 ml).

[0163] Clinical symptom observation is to observe general symptoms (whether there is blood in stool, diarrhea, etc.) and whether there is death / dying once a day for different individuals during the test. In this case, only individuals with specific symptoms are kept records (normal cases are not recorded). In addition, the body weight of the test animals is measured every day before purchase, grouping and administration of test substances.

[0164] In order to observe the immunohistochemical changes of colon tissue, the tissue fixed in 10% neutral buffered formalin solution was prepared into thin sections, attached to coating slides, deparaffinized and stained, and the immunohistochemistry (IHC) of antibodies interleukin-6 (IL-6), α-smooth muscle actin (α-SMA), and transforming growth factor-β (TGF-β) were respectively reacted on each section, and an external agency (Knotus) was commissioned to perform. Three designated areas of tissue sections of different individuals were designated and magnified 100 times to take pictures, and the stained part was measured compared to the total area and the average value was calculated (Zen 2.3 blueingtion, Carl Zeiss, Germany).

[0165] To induce ulcerative colitis, 2% sodium dextran sulfate was added to the drinking water of mice and continuously supplied for the first 7 days of each experimental period (3 weeks). The test substance was orally administered twice a day, 5 days a week for a total of 6 weeks together with the second sodium dextran sulfate induction. Fig.14 The dose of the test substance was converted according to the body weight in different individuals, and 20 mL of the drug was administered per kg of body weight. The above administration was performed by placing the test substance into a syringe and then connecting it to a rat feeding tube for oral administration.

[0166] During the trial, all dextran sulfate sodium DSS intake groups showed a significant increase in the ulcerative colitis disease activity index (DAI) characterized by significant weight loss and diarrhea after 7 days. After a 14-day recovery period, the disease activity index returned to near normal before the next dextran sulfate sodium induction cycle began. No significant differences in the disease activity index were observed between the test substance administration groups. In the changes in colon weight, length and colon proportion, an increase in colon weight, a decrease in colon length and an increase in colon proportion were observed in all dextran sulfate sodium intake groups. No significant differences in the above symptoms caused by the intake of dextran sulfate sodium were observed in the test substance administration groups.

[0167] In order to evaluate the therapeutic effect of the test substance on ulcerative colitis, after the end of the experiment, the animals were anesthetized and bled after laparotomy, and different individuals were subjected to histopathological examination. Histopathological findings are shown in Table 8 below, which are subdivided into changes in the severity of inflammation, crypt cell damage, ulcers, edema, and fibrosis and scored. In addition, after adding the severity scores of the above four histopathological studies except fibrosis, the positive control group (G2) was compared with the test substance administration group (G3, G4, G4).

[0168] [Table 8]

[0169]

[0170] Statistical analysis was performed using Graphpad Prism 8.1, with p < 0.05 as the minimum unit of significance. All tissue pathological indices measured were non-parametric data, and were represented in the curve graph by the median value, 25-75% value (in the box), and minimum-maximum value (two extremes) ( Figures 15 to 20 ). The Kruskal-Wallis test, which is a nonparametric test method, was used for variance analysis (ANOVA), and the Dunn's test was used to test the statistical significance between the positive control group (▲ represents the G2 group) and the test substance administration group (◇ represents the G3 group, ● represents the G4 group, and ◆ represents the G5 group). And the average values ​​are shown in Table 9.

[0171] [Table 9]

[0172]

[0173] The results of measuring the IL-6 staining coloration area and calculating the mean and standard deviation showed that the G2 group observed a 15.8-fold increase in area compared to the G1 group, showing statistical significance (p<0.01). Compared to the G2 group, the test substance administration groups (G3 to G5 groups) showed a decrease of 8.7%, 23.0% and 56.3% in area, respectively. In particular, the G5 group showed a significant decrease compared to the G2 group (Table 10, Fig.21 , Fig. 22 ).

[0174] [Table 10]

[0175] Experimental Group Interleukin 6 (area %) G1 0.8±0.3 G2 <![CDATA[12.6±4.2 ** ]]> G3 11.5±1.9 G4 9.7±2.9 G5 <![CDATA[5.5±1.5 ## ]]> .

Claims

1. A controlled-release composition for oral administration, characterized in that: A composite comprising a compound of Chemical Formula 1 or a pharmaceutically acceptable salt thereof and bentonite, The weight of the bentonite is 5-50 times the weight of the compound of Chemical Formula 1, The peak time of blood drug concentration (t max ) is longer than the time to peak blood concentration (t max ), The peak drug concentration (C max ) is lower than the peak drug concentration (C max ), The absorption rate of the compound of Chemical Formula 1 in the small intestine when the above composition is orally administered is slower than the absorption rate of the compound of Chemical Formula 1 in the small intestine when an aqueous solution is orally administered. [Chemical formula 1] 2. The controlled-release composition for oral administration according to claim 1, characterized in that: The peak time of blood drug concentration (t max ) is shorter than the time to peak blood concentration (t max ) 2 to 10 times longer.

3. The controlled-release composition for oral administration according to claim 1, characterized in that: The peak drug concentration (C max ) is the peak drug concentration (C max ) from 20% to 80%.

4. The controlled-release composition for oral administration according to claim 1, characterized in that: The drug improving effect of the compound of Chemical Formula 1 on the symptoms selected from the group consisting of inflammation, crypt cell damage, ulcer, edema and fibrosis in the colon when the above composition is orally administered is better than the drug improving effect of the compound of Chemical Formula 1 in the colon when an aqueous solution is orally administered.

5. The controlled-release composition for oral administration according to claim 1, characterized in that: The dissolution rate of the above complex when placed in an eluent having a pH of 1.2 is slower than the dissolution rate of the compound of Chemical Formula 1 when placed in an eluent having a pH of 1.

2.

6. The controlled-release composition for oral administration according to claim 1, characterized in that: The dissolution rate of the above complex when placed in an eluent having a pH of 7.4 is faster than the dissolution rate of the compound of Chemical Formula 1 when placed in an eluent having a pH of 7.

4.

7. A method for preparing a controlled-release composition for oral administration, characterized in that: The method comprises the steps of mixing a solution containing a compound of Chemical Formula 1 or a pharmaceutically acceptable salt thereof, an acidic reaction solution and a bentonite suspension and adsorbing the compound onto the bentonite to prepare a composite. The controlled-release composition for oral administration comprises a composite of the compound of Chemical Formula 1 and bentonite, The weight of the bentonite is 5-50 times the weight of the compound of Chemical Formula 1, The peak time of blood drug concentration (t max ) is longer than the time to peak blood concentration (t max ), The peak drug concentration (C max ) is lower than the peak drug concentration (C max ), The absorption rate of the compound of Chemical Formula 1 in the small intestine when the above composition is orally administered is slower than the absorption rate of the compound of Chemical Formula 1 in the small intestine when an aqueous solution is orally administered. [Chemical formula 1] 8. The method for preparing the controlled-release composition for oral administration according to claim 7, characterized in that: The pH value of the above reaction solution is pH 0.5 to pH 3.5.

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