Temperature sensitive adhesion prevention composition and use thereof

Abstract

Provided are a temperature sensitive adhesion prevention composition and a method for producing the same, the composition comprising a polyethylene oxide (PEO) containing copolymer, a carboxy polysaccharide, and a monovalent cation.

Claims

1. An adhesion prevention composition comprising a polyethylene oxide(PEO)-comprising copolymer, a carboxypolysaccharide, and a monovalent cation, wherein the composition is capable of transitioning from sol state to gel state upon raising temperature and does not contain a multivalent cation.

2. The adhesion prevention composition of claim 1, wherein the monovalent cation is one or more selected from the group consisting of Li+, Rb+, Fr+, Na+, NO2+, NH4+, H30+, and K+.

3. The adhesion prevention composition of claim 1, wherein the copolymer comprises polyethylene oxide in an amount of 10% by weight to 90% by weight.

4. The adhesion prevention composition of claim 1, wherein the copolymer comprises another component which is copolymerized with the polyethylene oxide, another component being one or more selected from the group consisting of polypropylene oxide (PPO), polylactic acid (PLA), polyglycolic acid (PGA), poly(DL-lactide-co-glycolide) (PLGA), polycaprolactone (PCL), and poly(dioxanone) (PDO).

5. The adhesion prevention composition of claim 1, wherein the carboxypolysaccharide is one or more selected from the group consisting of hyaluronic acid, alginic acid, and carboxymethylcellulose.

6. The adhesion prevention composition of claim 1, wherein a weight ratio of the copolymer to the composition is 0.1:1 to 0.4:1.

7. The adhesion prevention composition of claim 1, wherein a weight ratio of the carboxypolysaccharide to the composition is 0.001:1 to 0.03:1.

8. A method for producing an adhesion prevention composition, the method comprising providing a polyethylene oxide(PEO)-comprising copolymer, a carboxypolysaccharide, and a monovalent cation and forming a composition, wherein the composition is capable of transitioning from sol state to gel state upon raising temperature and does not contain a multivalent cation.

9. The method of claim 8, wherein the monovalent cation is one or more selected from the group consisting of Li+, Rb+, Fr+, Na+, NO2+, NH4+, H30+, and K+.

10. The method of claim 8, wherein the copolymer comprises polyethylene oxide in an amount of 10% by weight to 90% by weight.

11. The method of claim 8, wherein the copolymer comprises another component which is copolymerized with the polyethylene oxide, another component being one or more selected from the group consisting of polypropylene oxide (PPO), polylactic acid (PLA), polyglycolic acid (PGA), poly(DL-lactide-co-glycolide) (PLGA), polycaprolactone (PCL), and poly(dioxanone) (PDO).

12. The method of claim 8, wherein the carboxypolysaccharide is one or more selected from the group consisting of hyaluronic acid, alginic acid, and carboxymethylcellulose.

13. The method of claim 1, wherein a weight ratio of the copolymer to the composition is 0.1:1 to 0.4:1.

14. The method of claim 1, wherein a weight ratio of the carboxypolysaccharide to the composition is 0.001:1 to 0.03:1.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

(1) FIG. 1 shows the result of evaluating tissue adhesions using an adhesion test system according to 4 grades of tissue adhesion severity (if the score of 0, 1, 2, or 3 is higher, adhesion is more severe).

EXAMPLES

(2) Hereinafter, the present invention will be described in more detail with reference to Examples. However, these Examples are for illustrative purposes only, and the scope of the present invention is not intended to be limited by these Examples.

Examples 1-5: Preparation of Temperature-Sensitive Adhesion Prevention Composition

(3) A thermosensitive composition was prepared using components and contents as described in the following Table 1 as raw materials. In detail, sodium chloride (NaCl) and hyaluronic acid (HA) were serially dissolved in an ultrapure water, and stirred using a homogenizer. Then, poloxamers (poloxamer 188/poloxamer 407, P188/407 at a weight ratio of (4/6)) were added thereto at a predetermined ratio and content, and dissolved at 20 C. or lower using the homogenizer to prepare a thermosensitive composition solution including poloxamer, hyaluronic acid, and sodium chloride.

(4) TABLE-US-00001 TABLE 1 Component Content (w/w %) Example 1 Example 2 Example 3 Example 4 Example 5 Hyaluronic 0.5 0.5 0.5 0.5 0.5 acid Poloxamer 32 (4/6) 32 (4/6) 32 (4/6) 32 (4/6) 32 (4/6) (188/407) NaCl 2 1.5 1 0.5 0.25

Comparative Examples 1-5: Preparation of Temperature-Sensitive Adhesion Prevention Composition

(5) A thermosensitive composition was prepared in the same manner as in Examples 1-5, except that each component was used in an amount shown in the following Table 2.

(6) TABLE-US-00002 TABLE 2 Content Compar- Compar- Compar- Compar- Compar- Component ative ative ative ative ative (w/w %) Example 1 Example 2 Example 3 Example 4 Example 5 Hyaluronic 0.5 0.5 0 0.5 0 acid Poloxamer 32 (4/6) 0 32 (4/6) 0 32 (4/6) (188/407) NaCl 0 0.5 0.5 0 0

Comparative Examples 6: Preparation of Temperature-Sensitive Adhesion Prevention Composition Using Multivalent Ions

(7) A thermosensitive composition was prepared in the same manner as in Examples 1-5, except that each component was used in an amount shown in the following Table 3.

(8) TABLE-US-00003 TABLE 3 Content Compar- Compar- Compar- Compar- Compar- ative Component ative ative ative ative Example (w/w %) Example 6 Example 7 Example 8 Example 9 10 Hyaluronic 0.5 0.5 0.5 0.5 0.5 acid Poloxamer 32 (4/6) 32 (4/6) 32 (4/6) 32 (4/6) 32 (4/6) (188/407) CaCl.sub.2 1 0.5 0.1 0.05 0.01

Examples 6-9: Preparation of Temperature-Sensitive Adhesion Prevention Composition

(9) A thermosensitive composition was prepared in the same manner as in Examples 1-5, except that each component was used in an amount shown in the following Table 4.

(10) TABLE-US-00004 TABLE 4 Component Content (w/w %) Example 6 Example 7 Example 4 Example 8 Example 9 Hyaluronic 0.5 0.5 0.5 0.5 0.5 acid Poloxamer 9 (4/6) 20 (4/6) 32 (4/6) 40 (4/6) 50 (4/6) (188/407) NaCl 0.5 0.5 0.5 0.5 0.5

Experimental Example 1: Observation of Precipitation of Temperature-Sensitive Adhesion Prevention Composition

(11) The temperature-sensitive adhesion prevention compositions prepared in Examples 1-5 and Comparative Examples 1-5 were observed for precipitation and the results are shown in Table 6.

(12) As shown in Table 3, no precipitation was observed in the compositions of Examples 3 to 5, that is, each composition including NaCl of 1% by weight, 0.5% by weight, or 0.25% by weight, hyaluronic acid, and poloxamer. In contrast, precipitation was observed in the compositions of Examples 1 and 2, that is, each composition including NaCl of 2% by weight or 1.5% by weight, hyaluronic acid, and poloxamer. These results suggest that when a mixing ratio of NaCl is more than 1.5% by weight in a composition including hyaluronic acid, poloxamer, and NaCl, strong inter-hydrogen bonds between hyaluronic acid and poloxamer, and strong intra-hydrogen bonds inside hyaluronic acid and poloxamer are formed, causing precipitation of some of the compositions.

(13) TABLE-US-00005 TABLE 6 Presence or absence Presence or absence of precipitation of precipitation Example 1 Presence Comparative Absence Example 1 Example 2 Presence Comparative Absence Example 2 Example 3 Absence Comparative Absence Example 3 Example 4 Absence Comparative Absence Example 4 Example 5 Absence Comparative Absence Example 5

Experimental Example 2: Measurement of Gel Residence Time

(14) Each composition prepared in Examples 1-5 and Comparative Examples 1-5 was added to a 1-ml vial, and 1 ml of a phosphate buffer solution (pH 7.4) was added thereto. Then, the phosphate buffer solution on the surface layer of the gel was removed at a predetermined time once a day while storing the vials in an incubator at 37 C., and the residual volume was observed to measure a gel residence time. The results are shown in Table 7.

(15) As shown in Table 7, gel residence times of Examples 3 to 5 were 14 days and 10 days, which were longer than those of Comparative Examples.

(16) Example 1 and Example 2 were excluded from observation, because of precipitation. In the case of Examples 3 to 5, % by weight of each of the solutions containing monovalent cations was 1, 0.5, and 0.25 w/w %. In the case of Comparative Example 1 containing no NaCl, the gel residence time was 8 days, which was shorter than those of Examples 3 to 5. Therefore, the solutions containing cations of 0.1% by weight to less than 1.5% by weight were found to improve the gel residence time of the composition.

(17) These results suggest that the polyethylene oxide-containing copolymer/carboxypolysaccharide compositions having a particular content of monovalent cations increase hydrogen bonds inside the copolymer, hydrogen bonds inside the carboxypolysaccharide, and hydrogen bonds between the copolymer and the carboxypolysaccharide, thereby improving stability of the composition.

(18) Further, referring to Comparative Example 3 and Examples 3 to 5, it was confirmed that presence of a hydroxy polysaccharide such as hyaluronic acid further increases the gel residence time of the composition.

(19) TABLE-US-00006 TABLE 7 Gel residence time Gel residence time (day) (day) Example 1 Comparative 8 Example 1 Example 2 Comparative 2 Example 2 Example 3 14 Comparative 6 Example 3 Example 4 14 Comparative 1 Example 4 Example 5 10 Comparative 5 Example 5

Experimental Example 3: Test of Precipitation and Gel Residence Time of Temperature-Sensitive Adhesion Prevention Composition Using Multivalent Cations

(20) Precipitation and gel residence time of the multivalent cation-containing compositions prepared in Comparative Examples 6-10 were examined and the results are shown in Table 8.

(21) As shown in Table 8, precipitation of the compositions of Comparative Examples 6 to 10 was observed, that is, each composition including CaCl2 of 1% by weight, 0.5% by weight, 0.1% by weight, or 0.05% by weight, hyaluronic acid, and poloxamer. In contrast, no precipitation of the composition of Comparative Example 10 was observed, that is, the composition including CaCl2 of 0.01% by weight, hyaluronic acid, and poloxamer. The composition of Comparative Example 10 showing no precipitation was observed according to the method of measuring the gel residence time as in Experimental Example 2, and as a result, the compositions of Comparative Example 6-10 showed gel residence times similar to that of the composition of Example 6 containing no monovalent cation. These results suggest that use of the divalent cation CaCl2 did not improve the gel residence time. It was confirmed that the monovalent cation of the present invention effectively improves binding ability between the polyethylene oxide-containing copolymer and the carboxypolysaccharide polymer, thereby increasing the gel stability.

(22) TABLE-US-00007 TABLE 8 Presence or absence of precipitation Gel residence time Comparative Example 6 Presence Comparative Example 7 Presence Comparative Example 8 Presence Comparative Example 9 Presence Comparative Example 10 Absence 8

Experimental Example 4: Measurement of Sol-Gel Phase Transition Temperature

(23) Sol-gel transition temperatures of the compositions prepared in Examples 4 and 6-9 were measured. Each composition was added to a vial, the vials were stored in a refrigerator at about 4 C. for 30 minutes, and then each sample in a sol state was added to a 50-ml vial. The vials were placed in a circulating water bath, and a temperature at which viscosity of the composition was rapidly changed, that is, a temperature at which flowability of the composition disappeared while a temperature of the water bath was slowly raised was determined as a lower critical solution temperature (LCST). Each composition prepared in Examples 4 and 6-9 was measured in the same manner and the results are shown in Table 9.

(24) As shown in Table 9, it was confirmed that the composition of Example 6, that is, the composition including a polyethylene oxide-containing copolymer, which is a poloxamer, in an amount of 0.09% by weight or less, based on 1% by weight of the final composition, showed no sol-gel transition. It was also confirmed that the composition of Example 8, that is, the composition including the polyethylene oxide-containing copolymer, which is a poloxamer, in an amount of 0.4% by weight or more, based on 1% by weight of the final composition, showed sol-gel transition at a temperature lower than room temperature. Based on the results of Table 9, it can be seen that sol-gel phase transition occurs at a particular temperature according to the content of the polyethylene oxide-containing copolymer, which is a poloxamer having a property of temperature sensitivity.

(25) TABLE-US-00008 TABLE 9 Sol-gel phase transition temperature ( C.) Example 6 Example 7 60 Example 4 31 Example 8 10 Example 9

Experimental Example 5: Test of Adhesion Prevention Efficacy in Animals

(26) An animal test (cecum/abdominal wall abrasion rat model) was performed to evaluate tissue adhesion prevention efficacy, using a control group, in which no materials were added between the wound sites after administration of each composition of Example 3 and Comparative Examples 1-3 and 5.

(27) In the animal test, 5 SD rats (female) per group were used. First, ketamine (60100 mg/kg) and xylazine (510 mg/kg) were mixed and then the rats were anesthetized by injecting them with the mixture intraperitoneally. The abdominal hairs of the anesthetized rats were shaved and disinfected with povidone, and a 45 cm-long midline laparotomy was performed. The cecum was taken out and a wound of size 1 cm1 cm was formed in the surface of the cecal serosa using a bone burr until the serosal surface was hemorrhagic, and the adjacent abdominal wall was wounded using a 15T blade such that the epidermis was peeled off. The two abraded sites faced each other and no treatment was performed in the control group, and then the abdominal wall and skin layers were sutured successively. To the experimental groups, each prepared adhesion prevention composition (Example 3, Comparative Examples 1-3, 5) was uniformly applied to the abraded sites in the cecum and the abdominal wall, and then the abdominal wall and the skin layers were sutured successively.

(28) At 10 days after surgery, tissue adhesions were examined. The results of evaluating tissue adhesions using an adhesion test system according to 4 grades of tissue adhesion severity (if the score of 0, 1, 2, or 3 is higher, adhesion is more severe) are shown in FIG. 1. As shown in FIG. 1, it was confirmed that Example 3 showed remarkably excellent adhesion prevention efficacy, as compared with Comparative Examples 1-3, and 5. Further, as shown by the gel residence time results of Experimental Example 2, it was also confirmed that as the gel residence time increased, the adhesion prevention efficacy was more excellent. Accordingly, it was confirmed that the composition including poloxamer, which is a polyethylene oxide-containing copolymer, and hyaluronic acid, which is a kind of carboxypolysaccharide, together with a monovalent cation, from sodium chloride, showed a gel residence time of 10 days or longer, and also superior adhesion prevention efficacy in the animal test.