KR102745034B1 - Water-sensitive sol-gel phase transition-based anti-adhesion composition - Google Patents

Abstract

The present invention relates to a method for producing an anti-adhesion composition based on a liquid crystal structure having sol-gel phase transition characteristics, and to a composition produced therefrom. More specifically, the present invention is a novel anti-adhesion agent having a sol-gel phase transition mechanism that absorbs a body fluid when exposed to water or a body fluid. In a sol state, it has good flowability and is easy to apply, and in the body, it is converted into a gel form and can increase the persistence in the body through the effect of maintaining a physical barrier. By using a combination of compositions having biocompatibility, an anti-adhesion composition based on a liquid crystal structure that is decomposed, absorbed, and excreted and exhibits excellent anti-adhesion ability can be provided.

Description

{Water-sensitive sol-gel phase transition-based anti-adhesion composition}

The present invention relates to an anti-adhesion composition based on a liquid crystal structure having sol-gel phase transition characteristics when exposed to a biological fluid. Specifically, the present invention relates to an anti-adhesion composition based on a liquid crystal structure which significantly increases mechanical durability by gelling the composition using the sol-gel phase transition characteristics after absorbing peritoneal fluid in the body, thereby acting as a physical barrier.

Adhesion is a phenomenon in which tissues or membranes inside the human body stick together during the recovery process of damaged organs and tissues due to resection, suturing, physical damage, or incomplete hemostasis after surgery. The incidence of adhesion in intra-abdominal surgery or gynecological surgery is high, with 60-93% of cases occurring between the patient's abdominal wall and intestines, depending on the surgery.

Depending on the surgical site, the occurrence of adhesions can cause chronic pelvic pain, small bowel obstruction, stiffness due to joint adhesions, or restricted movement. If the symptoms are severe, reoperation for adhesion release is required. The occurrence of adhesions can extend the surgical time and affect the surgical prognosis due to the removal process of adhesions when performing other surgeries over time, so it is very important to use an appropriate anti-adhesion agent.

Most of the anti-adhesion agents commercialized in Korea are products using hyaluronic acid and sodium-carboxyl methyl cellulose, or are released in the form of gels that are attached to the surgical site by utilizing the sol-gel phase transition characteristics of thermosensitive poloxamer. However, although liquid anti-adhesion agents have excellent biocompatibility due to the characteristics of hyaluronic acid, their bioabsorption rate is fast and they are easily decomposed, so their anti-adhesion effect is limited.

In addition, there are cases where the formulation is detached from the application site due to movement of the organ after attachment, and it is difficult to use locally at a wound site because the adhesiveness is poor. However, the adhesion prevention composition based on the liquid crystal structure has sol-gel phase transition characteristics, so it has good flowability before injection into the body and excellent applicability, and it spreads thinly after application, so it is good for local use. Not only did we use a combination of compositions that have biocompatibility, but we also made it possible to form a physical barrier with higher gel viscoelasticity after the sol-gel phase transition than the sol-gel phase transition product composed of poloxamer.

Conventional temperature-sensitive adhesion-preventing hydrogels using hyaluronic acid derivatives as adhesion-preventing agents have attempted to improve the duration of hyaluronic acid in the body by using acrylamide-hyaluronic acid derivatives that are harmless to the human body. Although acrylamide-hyaluronic acid derivatives have the characteristic of stronger persistence in the body than existing hyaluronic acid derivatives, the present invention is significantly different in form in that it is moisture-sensitive in that gelation occurs due to moisture in the body and that it is an oil-based sol form as a liquid crystal precursor rather than a hydrogel, and thus has better persistence in the body. (Patent Document 0001)

In addition, previous studies using vegetable oils such as soybean oil, canola oil, and argan oil as anti-adhesion agents have presented results proving the effectiveness of oils in preventing adhesion, and soybean oil, canola oil, etc. are oils that have been approved for use as additives in medicines administered by intravenous and intramuscular injection, and have been proven to be safe for use in the body. However, oils have the disadvantage of not adhering to the surgical site and tend to flow down, making it difficult to apply to the desired site. (Patent Document 0002)

Phospholipids, the main component of biomembranes, have attracted the attention of researchers as substances that naturally exist in the peritoneal cavity. Phospholipids isolated from the peritoneum have been found to have the ability to reduce surface tension and promote water repellency. Here, they are known to act as a liquid barrier that separates appropriate areas of the peritoneal surface. In addition, there are results showing that phospholipids reduce profibrotic mediators released after surgical trauma after abdominal surgery and significantly reduce peritoneal adhesion formation. Phospholipids alone cannot remain well in the wound site and thus cannot exhibit an anti-adhesion effect, and the same goes for oils. Phospholipids include natural and synthetic phospholipids. Natural phospholipids are extracted from phospholipids contained in egg yolk, soybeans, etc. and are called lecithin. Lecithin may contain neutral lipids, polysaccharides, and cholesterol other than phospholipids, but phospholipid content accounts for the majority. The phospholipids contained in lecithin are mixed with different fatty acid chain lengths, degrees of unsaturation, and types of hydrophilic functional groups, but the purity of the total phospholipids can reach 40 to 99%. Synthetic phospholipids are phospholipids with specific chain lengths, degrees of unsaturation, and hydrophilic functional groups. (Patent Document 0003, Non-Patent Document 0001) The anti-adhesion composition according to the present invention can be produced not only with natural phospholipids but also with synthetic phospholipids.

However, the inventors of the present invention have discovered an excellent adhesion-prevention effect by using a composition for preventing adhesion with enhanced adhesiveness and physical strength, manufactured by optimizing phospholipids, auxiliary solvents, oils, and fatty acids, to effectively inhibit adhesion by staying at a wound site for a long period of time using sol-gel phase transition characteristics, thereby completing the present invention.

Korean Patent Publication No. 10-1871930 (Registration date: July 2, 2018) Korean Patent Publication No. 10-2388506 (Registration date April 15, 2022) Korean Patent Publication No. 10-0446833 (Registration date: August 24, 2004)

Fotiadis K, Filidou E, Arvanitidis K, Valatas V, Stavrou G, Basdanis G, Paspaliaris V, Kolios G, Kotzampassi K. Intraperitoneal application of phospholipids for the prevention of postoperative adhesions: a possible role of myofibroblasts. J Surg Res, 8, 295-297 (2015)

In order to solve the problem of effectively inhibiting adhesion by staying at a wound site for a long time, the present invention aims to provide an adhesion-prevention composition with excellent biocompatibility by using an amphipathic lipid, an oil or a fatty acid, and a biocompatible auxiliary solvent which form a liquid crystal structure when exposed to a biological fluid such as water, salt water or peritoneal fluid. The adhesion-prevention composition of the present invention is a composition that reduces the formation of peritoneal adhesions after abdominal surgery and increases adhesiveness and physical strength, and significantly complements the shortcomings of conventional adhesion-prevention agents by utilizing sol-gel phase transition characteristics.

The anti-adhesion composition according to the present invention relates to an anti-adhesion composition characterized in that it comprises an amphipathic lipid forming a liquid crystal; and a group consisting of oils or fatty acids derived from animals, plants or minerals, or a combination thereof.

The anti-adhesion composition of the present invention comprises a liquid crystal forming agent including an amphoteric lipid and a medium-chain fatty acid, and provides a composition that spontaneously forms liquid crystals in an aqueous medium.

In the anti-adhesion composition of the present invention, the amphiphilic lipid may be selected from the group consisting of phospholipids, glyceryl monooleate, glyceryl dioleate or glyceryl phytanoate, phytantriol, tocopherol acetate, and sorbitan unsaturated fatty acid ester.

In the anti-adhesion composition of the present invention, oils derived from animals, plants, and minerals may be selected from the group consisting of squalene, lanolin, mineral oil, paraffin, petroleum jelly, almond oil, corn oil, cottonseed oil, olive oil, peanut oil, safflower seed oil, grape seed oil, sesame oil, soybean oil, argan oil, canola oil, castor oil, coconut oil, sunflower seed oil, palm oil, avocado oil, and medium-chain triglycerides.

In the anti-adhesion composition of the present invention, the fatty acid may be selected from at least one selected from the group consisting of caproic acid, caprylic acid, capric acid, myristoleic acid, palmitoleic acid, oleic acid, linoleic acid, linolenic acid, arachidonic acid, eicosapentaenoic acid, eruic acid, and docosahexanoic acid.

The anti-adhesion composition of the present invention relates to an anti-adhesion composition characterized by comprising: an amphipathic lipid forming a liquid crystal; a group consisting of oils or fatty acids derived from animals, plants or minerals, or a combination thereof; and a biocompatible auxiliary solvent.

The biocompatible auxiliary solvent corresponding to the optional component added in the anti-adhesion composition of the present invention may be selected from one or more of ethanol, glycerin, propylene glycol, polyethylene glycol, N-methyl-2-pyrrolidone, and propylene carbonate.

The anti-adhesion composition of the present invention may contain 20 to 90 wt% of amphiphilic lipids based on the total weight, and preferably 25 to 90 wt% of amphiphilic lipids based on the total weight of the anti-adhesion composition.

The anti-adhesion composition of the present invention may contain 0.1 to 75 wt% of oil and/or fatty acid derived from animals, plants and minerals, based on the total weight, and preferably 10 to 75 wt%.

The anti-adhesion composition of the present invention may contain 0 to 35 wt% of a biocompatible auxiliary solvent based on the total weight.

Amphiphilic lipids are formulations that induce a sol-gel phase transition through solvent exchange, and form a gel in the form of a liquid crystal. Since a liquid crystal former composed of an amphiphilic lipid spontaneously forms a liquid crystal with a thermodynamically stable structure in the peritoneum, the sol changes into a gel form. At this time, the sol form has low viscosity so that it can be easily applied, and when it changes into a gel form, it has high viscosity and adhesiveness so that it can adhere to the peritoneum for a long time. The present invention is not limited to a specific surgical technique as a sol-gel phase-transition formulation and can be used in various ways. In addition, the following examples are illustrative of the present invention, and the present invention is not limited to the examples.

Figure 1 shows that Comparative Example 3 is a formulation made only with a combination of an amphiphilic lipid and a co-solvent, and Example 18 is a formulation made with a combination of an amphiphilic lipid, a co-solvent, and a fatty acid. When the formulations were put in a petri dish and tilted, Comparative Example 3 fell immediately due to gravity, indicating that it had low viscosity, and Example 18 was confirmed to stay at the application site, confirming that the retention at the adhesion site was enhanced when the formulation was used as an anti-adhesion agent.
Figure 2 shows the form of an anti-adhesion agent before and after contact with an aqueous medium in an embodiment and a comparative example of the present invention.
Figure 3 is a photograph showing the results of the adhesion prevention efficacy test of the adhesion prevention compositions of the present invention based on the examples and control examples of the present invention at autopsy. The adhesions that occurred as the organs recovered are indicated by yellow dotted lines, and the adhesion scores are also recorded. In the case of control examples 1 and 2, it can be seen that strong adhesions occurred in all rats.

The present invention is described in detail below.

In the present invention, the term "adhesion" may refer to a phenomenon in which skin or membranes that are separated from each other stick to each other. When tissue damage occurs after surgery or due to inflammation, foreign bodies, bleeding, infection, wounds, friction, chemical treatment, etc., blood leaks out and coagulates during the wound healing process, and this causes abnormal adhesion with surrounding tissues. Such adhesion occurs frequently, especially after surgery, and when adhesion occurs in the pelvis, it can cause chronic pain, sexual dysfunction, etc., and after thyroid removal surgery, side effects such as chest pain and dysphagia occur due to adhesion, and when adhesion occurs due to spinal surgery, the nerves are compressed, causing severe pain, and it is known that adhesion in the uterus causes infertility, amenorrhea, and habitual miscarriage.

As used herein, the term “biological fluid” includes but is not limited to any liquid, treated or untreated, associated with a living organism, such as water, saline solution, peritoneal fluid, treated or untreated blood, nutrients and/or anticoagulant solutions, and as used herein, biological fluid means a similar biological fluid obtained by the aforementioned components and other methods and having similar properties.

The anti-adhesion composition of the present invention is an anti-adhesion composition that spontaneously forms liquid crystals in a biological fluid, and is characterized by comprising: an amphipathic lipid; a group consisting of oils or fatty acids derived from animals, plants or minerals, or a combination thereof; and/or a biocompatible auxiliary solvent.

The principle of the moisture-sensitive sol-gel phase transition anti-adhesion agent of the present invention is the principle of forming a liquid crystal gel with a high concentration of amphiphilic lipids as a matrix through solvent exchange when in contact with water to form a physical barrier. In the combination of amphiphilic lipids and co-solvents, the co-solvent can dissolve amphiphilic lipids well and form a gel when mixed with water. However, considering the capacity used in the anti-adhesion agent, the use of the co-solvent alone may be excessively high. In the combination of amphiphilic lipids and oils or fatty acids, the range in which phospholipids can be dissolved to the maximum is low. In addition, in the combination of amphiphilic lipids, oils or fatty acids, and co-solvents, reducing the co-solvent in the form of amphiphilic lipids and co-solvents can play a role in resolving toxicity issues, increasing homogeneity during sol-gel phase transition, and enhancing the viscosity of the formulation. (See Fig. 1)

The anti-adhesion composition of the present invention may include at least one selected from the group consisting of phospholipids, glyceryl monooleate, glyceryl dioleate or glyceryl phytanoate, phytantriol, phospholipids, tocopherol acetate, and sorbitan unsaturated fatty acid esters, which are amphiphilic lipids that spontaneously form liquid crystals in a biological fluid.

When the amphiphilic lipid of the present invention is less than 25 wt%, the amphiphilic lipid is too little to have a viscosity that is too low to prevent adhesion, and it is difficult to form a matrix, which is a physical barrier, when applied to the body. In addition, when the amphiphilic lipid exceeds 90 wt%, it is difficult for the amphiphilic lipid to be dissolved by the auxiliary solvent, and it is difficult for the formulation to have uniformity.

The anti-adhesion composition of the present invention can use oils and fatty acids derived from animals, plants, and minerals. The oil helps the formulation to last a long time when administered in the body in an oily form, and suppresses the occurrence of adhesions at wound sites in the abdominal cavity.

The oil derived from animals, plants or minerals of the present invention may include at least one selected from the group consisting of squalene, lanolin, mineral oil, paraffin, petrolatum, almond oil, corn oil, cottonseed oil, olive oil, peanut oil, safflower oil, grapeseed oil, sesame oil, soybean oil, argan oil, canola oil, castor oil, coconut oil, sunflower seed oil, palm oil, avocado oil and medium-chain triglycerides, and the fatty acid may include at least one selected from the group consisting of caproic acid, caprylic acid, capric acid, myristoleic acid, palmitoleic acid, oleic acid, linoleic acid, linolenic acid, arachidonic acid, eicosapentaenoic acid, eruic acid and docosahexanoic acid.

The fatty acids and oils derived from animals, plants or minerals of the present invention may range from 10 to 75 wt% based on the total weight. When the fatty acids and oils derived from animals and plants are less than 10 wt%, the amphipathic lipids do not dissolve well, and when they exceed 75 wt%, the formulation is not stable and phase separation occurs.

The anti-adhesion composition of the present invention may add a suitable auxiliary solvent for formulation formation. The auxiliary solvent may be selected from the group consisting of, but is not limited to, ethanol, glyceryl, propylene glycol, polyethylene glycol, N-methyl-2-pyrrolidone, propylene carbonate, and combinations thereof, for example.

The biocompatible auxiliary solvent of the present invention may be contained in an amount of 0 to 35 wt% based on the total weight. If the auxiliary solvent exceeds 35 wt%, toxicity problems may occur in cases of surgery in which an excessive amount of an anti-adhesion agent is used.

<Examples 1 to 12 and Comparative Examples 1 to 5> Preparation of liquid crystal forming composition containing ethyl alcohol

Comparative Examples 1 to 5 and Examples 1 to 12 containing lecithin (Lipoid E 80, Lipoid GmBH) were prepared according to the ingredients and contents of Table 1 below. The phospholipid used in the present invention is composed of lecithin including phosphatidyl choline at 80 wt% based on the total composition. Specifically, lecithin, a liquid crystal former and a medium-chain triglyceride as a liquid crystal structure formation assistant, and ethyl alcohol (Ethyl Alcohol, Duksan) were mixed in a glass vial while stirring with a magnetic stirrer at 60°C. The total batch size was prepared at 5 g per preparation.

The formulations corresponding to Table 1 below were all well-formed and also gelled well when exposed to an aqueous medium such as a biological fluid.

Since a high content of ethyl alcohol in a formulation containing ethyl alcohol may be harmful to the body, Table 1 was created by considering the weight % of ethyl alcohol. (See Figure 4)

As shown in Table 7, when the auxiliary solvent, ethyl alcohol, was 40 wt%, rats died, and when it was 40 wt% or more, ethyl alcohol showed harmful effects when administered to the body, so formulations exceeding 40 wt% were set as comparative examples.

type
(Unit: g) Comparative example Example 1 2 3 4 5 1 2 3 Phospholipid 1.5 2.5 3.5 1.5 2.5 3.5 1.5 2.5 Heavy chain tree
Glycerides - - - 0.7 0.5 0.5 1.75 1.25 ethyl alcohol 3.5 2.5 1.5 2.8 2.0 1 1.75 1.25 type
(Unit: g) Example 4 5 6 7 8 9 10 Phospholipid 3.5 1.5 2.5 3.5 1.5 2.5 3.5 Medium chain triglycerides 0.75 2.33 1.67 1 2.8 2 1.2 ethyl alcohol 0.75 1.17 0.83 0.5 0.7 0.5 0.3

<Examples 11 to 22> Preparation of a liquid crystal forming composition containing propylene glycol

Examples 11 to 22 containing phospholipids were prepared according to the ingredients and contents of Table 2 below. Specifically, lecithin (Lipoid E 80, Lipoid GmBH), a liquid crystal former and a liquid crystal structure forming aid, medium-chain triglyceride, and propylene glycol (Propylene glycol, Woonpoong Pharm) were mixed in a glass vial while stirring with a magnetic stirrer at 60°C. The total batch size was prepared as 5 g per preparation.

The formulations corresponding to Table 2 below were all well-formed and also gelled well when exposed to an aqueous medium such as a biological fluid. (See Figure 2)

Through Examples 11 to 22, it was shown that not only formulations using ethyl alcohol but also formulations using propylene glycol were well formed, and thus, an anti-adhesion composition can be prepared using other auxiliary solvents.

type
(Unit: g) Example 11 12 13 14 15 16 Phospholipid 3.5 2.5 3.5 1.5 2.5 3.5 Heavy chain tree
Glycerides 0.3 0.83 0.5 1.75 1.25 0.75 Propylene
Glycol 1.2 1.67 1 1.75 1.25 0.75 type
(Unit: g) Example 17 18 19 20 21 22 Phospholipid 1.5 2.5 3.5 1.5 2.5 3.5 Heavy chain tree
Glycerides 2.33 1.67 1 2.8 2 1.2 Propylene
Glycol 1.176 0.83 0.5 0.7 0.5 0.3

<Examples 23 to 28> Preparation of liquid crystal-forming compositions containing various phospholipids Examples 23 to 34 containing glyceryl monooleate (Glycerol Monoolate, Gattefosse), sorbitan monooleate (Span 80, Samchun), and tocopherol acetate (DL-α-Tocopherol acetate, Daejung) were prepared according to the ingredients and amounts shown in Table 3 below. Specifically, lecithin (Lipoid E 80, Lipoid GmBH), glyceryl monooleate as a liquid crystal forming agent, and propylene glycol (Propylene glycol, Woonpoong Pharm) or ethyl alcohol (Ethyl Alcohol, Duksan) were mixed in a glass vial while stirring with a magnetic stirrer at 60°C. The total batch size was prepared at 5 g per formulation.

The formulations corresponding to Table 3 below were all well-formed and also gelled well when exposed to an aqueous medium such as a biological fluid.

Through Examples 23 to 28, it was shown that a formulation was well formed even when glyceryl monooleate was used in the amphiphilic lipid phospholipid, and thus an anti-adhesion composition can be prepared using other amphiphilic lipids.

type
(Unit: g) Example 23 24 25 26 27 28 Phospholipid 3 3 3 3 3 3 Tocopherol acetate 0.5 0.5 - - - - Sorbitan monooleate - - 0.5 0.5 - - Glyceryl Monooleate - - - - 0.5 0.5 Medium chain triglycerides 1 1 1 1 1 1 Propylene glycol 0.5 - 0.5 - 0.5 - ethyl alcohol - 0.5 - 0.5 - 0.5

<Examples 29 to 46> Preparation of liquid crystal forming composition using fatty acid and oil

Examples 29 to 46 were prepared using fatty acids and oils according to the ingredients and amounts in Table 4 below and ethanol as a solvent. Specifically, one of the fatty acids and oils of phospholipid and capric acid, caprylic acid, caproic acid, soybean oil, canola oil, grape seed oil, and olive oil was selected into a glass vial, ethyl alcohol was added, and the mixture was mixed while stirring with a magnetic stirrer at 60°C. The total batch size was prepared as 5 g per preparation, and whether or not the formulation was formed was indicated. The composition of the present invention creates a suitable liquid crystal-forming composition without separation of substances in the presence of an auxiliary solvent.

Through Examples 29 to 46, it was shown that formulations were well formed even when using fatty acids and oils, and thus an anti-adhesion composition can be prepared using other fatty acids or oils.

type
(Unit: g) Example 29 30 31 32 Phospholipid 2.5 1.25 3.75 3.75 Capric acid 2.5 - 0.625 - Soybean oil - 3.75 - 0.625 Ethyl alcohol - - 0.625 0.625 Whether the formulation is formed O O O O type
(Unit: g) Example 33 34 35 36 37 38 39 40 Phospholipid 2.5 2.5 2.5 2.5 3.75 3.75 3.75 3.75 Caproic acid 2.5 - - - 0.625 - - - Caprylic acid - 2.5 - - - 0.625 - - Oleic acid - - 2.5 - - - 0.625 - Linoleic acid - - - 2.5 - - - 0.625 Ethanol - - - - 0.625 0.625 0.625 0.625 Whether the formulation is formed O O O O O O O O type
(Unit: g) Example 41 42 43 44 45 46 Phospholipid 1.25 1.25 1.25 3.75 3.75 3.75 Canola oil 3.75 - - 0.625 - - Grapeseed oil - 3.75 - - 0.625 - olive oil - - 3.75 - - 0.625 Ethanol - - - 0.625 0.625 0.625 Whether the formulation is formed O O O O O O

< Experimental Example 1> Formulation experiment of a composition containing 25 to 90 wt% of lipid and confirmation of sol-gel phase transition The compositions made in the present invention are liquid crystal formulations that exhibit a sol-gel phase transition when exposed to an aqueous medium.

Therefore, after creating a composition containing 25 to 90 wt% of phospholipids, the sol-gel phase transition change was confirmed by starting to add a small amount of water. In addition, Examples 9, 21, 22, 29, 30, 31, and 32 were selected among the examples to confirm the sol-gel phase transition. The above combinations were selected to confirm that the sol-gel phase transition change occurred despite the differences in the concentration and type of the auxiliary solvent, the concentration of the amphiphilic lipid, and the concentration and type of the fatty acid and oil. (See Fig. 2)

type
(Unit: g) Example 9 21 22 29 30 31 32 Phospholipid 2.5 2.5 3.5 2.5 1.25 3.75 3.75 Heavy chain tree
Glycerides 2 2 1.2 - - - - Propylene
Glycol - 0.5 0.3 - - - - Ethyl alcohol 0.5 - - - - 0.625 0.625 Capric acid - - - 2.5 - 0.625 - Soybean oil - - - - 3.75 - 0.625 Sol-gel phase transition
Whether or not there is a change O O O O O O O

When the formulation was shaken after adding 1 ml of water, Examples 9, 21, 22, 29, 30, 31, and 32 showed a sol-gel phase transition, and Comparative Examples 6, 7, 8, and 9 (Table 6) remained in a sol state and had high flowability when in contact with an aqueous medium similar to a biological fluid, so it was confirmed that they had low viscosity when administered into the body, making them less effective in preventing adhesion. (Figure 2)

type
(Unit: g) Comparative Example 6 Comparative Example 7 Comparative Example 8 Comparative Example 9 Phospholipid 0.25 0.5 0.25 1.0 Medium chain triglycerides 2.375 2.25 2.375 2.5 Propylene glycol - - 2.375 1.5 ethyl alcohol 2.375 2.25 - - Whether the sol-gel phase transition changes X X X X

Comparative Examples 6 to 9 are formulations in which the concentration of amphiphilic lipid is 20 wt% or less, and as shown in Table 6, it was confirmed that an auxiliary solvent is required to make a liquid crystal-forming composition according to the present invention, and that the sol-gel phase transition change occurs from a suitable concentration range of 25 wt% or more of phospholipid. (Figure 2)

Additionally, the change in viscosity when in contact with an aqueous medium was shown in the drawing. It was confirmed that when the concentration of phospholipids is less than 20 wt%, no sol-gel phase transition change occurs and a low viscosity is maintained when in contact with an aqueous medium.

<Experimental Example 2> Experiment to confirm adhesion prevention ability of a composition containing amphipathic lipids using a rat peritoneal ligation model

In order to evaluate the adhesion prevention ability of Comparative Example 5, Examples 9, 21, 22, 29, 30, 31, and 32, an in vivo adhesion prevention ability evaluation experiment was conducted using 8-week-old male Sprague Dawley rats (Laon Bio, 250 g) as the experimental subjects. The experimental control group used rats that were not treated at all (corresponding to Control Example 1) and Gadix-sol (corresponding to Control Example 2). The experimental group used Examples 9, 21, 22, 29, 30, 31, 32, and Comparative Example 5 as adhesion prevention agents.

The number of animals used in the experiment was 3 (n = 3) for both the control group and the experimental group. The experiment was conducted as follows. (Figure 3)

After purchasing the laboratory animals, they are separated into experimental and control groups after a one-week acclimatization period.

Sedation and anesthesia are achieved using a small anesthesia machine using Isoflurane (Anesthesia, 3%, Inhalation), an animal anesthetic.

Once anesthesia has been administered, the abdominal incisions are shaved.

SD rats with shaved hair were moved to the operating table, the abdominal area was disinfected with povidone-iodine, and a 4 cm abdominal incision was made using a scalpel in the center of the abdomen.

To induce adhesion formation, a 3*3 wound is made in the peritoneum using a biopsy punch and the tissue is cut out.

For the control group, no composition was applied to the wound area, and for the experimental group, 0.5 ml of the composition was applied to the wound area.

After applying the composition, the incision site is closed by suturing it with surgical suture.

Before the rat wakes up from anesthesia, lay it down so that the wound area does not touch the floor.

After waking up from anesthesia, monitor the patient's condition and observe clinical symptoms daily.

After raising the animals for 10 days with sufficient food and water, they were euthanized by inhaling carbon dioxide gas and the degree of adhesion was assessed.

After 10 days of rearing of the control group that did not apply an anti-adhesion agent and the experimental group that applied compositions using phospholipids, the animals were re-ablated to confirm the efficacy of adhesion prevention. The degree of adhesion formation was visually confirmed and classified into 4 grades. “Grade 0 = No adhesions”, “Grade 1 = Thin adhesions that can be easily separated without much force”, “Grade 2 = Thick adhesions that require some force to be pulled tautly to separate”, “Grade 3 = Thick adhesions that cannot be separated without scissors and pulling force is insufficient”, “Grade 4 = Complex adhesions formed by intermixing with intra-abdominal organs” were each classified and determined.

Test group No.1 No.2 No.3 Adhesion assessment average score (n=3) Example 9 0 0 0 0 Example 21 0 1 0 0.33 Example 22 0 0 1 0.33 Example 29 0 0 0 0 Example 30 0 0 0 0 Example 31 0 1 0 0.33 Example 32 0 1 0 0.33 Comparative Example 5 0 5 dying 2.5 (N=2) Contrast example 1 4 2 3 3.67 Contrast example 2 2 3 1 2

Looking at Table 7, Comparative Example 5, in which the rat died, had an ethanol weight % of 40 wt%, and Example 9, in which the ethanol weight % was 10 wt%, showed no adhesion and no rats died, indicating that even in a composition containing ethanol, if the weight % of ethanol is lowered by adding medium-chain triglycerides, adhesion does not occur and there is no harmful effect on the rats. Based on the above results, the maximum weight % of ethanol, a type of auxiliary solvent that dissolves phospholipids well, is 35 wt%.

In addition, when the adhesion score was confirmed compared to the commercial product (control example 2), the average adhesion evaluation scores of Examples 9, 21, 22, 29, 30, 31, and 32 of the present invention were lower than those of the control examples, confirming that adhesion can be effectively prevented compared to commercial products. (Figure 3)

The example in Fig. 2 is an example in which the content of amphoteric lipids (phospholipids), which are the formers of liquid crystals, is 25 wt% or more of the total weight, includes oils and a biocompatible auxiliary solvent, and a sol-gel phase transition is confirmed. It is confirmed that the formation of a physical barrier due to the sol-gel phase transition and the wound healing effect of phospholipids have a good effect on preventing adhesion (since it is not valid to distinguish between 0 and 0.33 points, it is summarized as ‘good adhesion prevention effect’). However, in the case of the comparative example, when too much biocompatible auxiliary solvent was included (ethanol, weight ratio 40%), toxicity occurred and one rat died, and one of the remaining two rats was confirmed to have aggravated adhesion due to multiple inflammatory reactions. In the untreated group of Control Example 1, serious adhesion was found in all rats, and the group using Gadix sol (Control Example 2) showed a better effect than Control Example 1, but still did not show a sufficient adhesion prevention effect.

Specifically, Example 21 of the present invention was confirmed to have an anti-adhesion effect by comparing it with a commercially available anti-adhesion agent (Control Example 2) in a formulation mixed with propylene glycol, phospholipids, and medium-chain triglycerides. The results of an animal experiment on adhesion prevention by visual observation through autopsy are shown in Table 7, and the anti-adhesion agent of the present invention showed a superior anti-adhesion effect in terms of the degree of adhesion measured based on adhesion strength and adhesion area compared to the commercially available anti-adhesion agent. (See Fig. 3)

Claims (7)

In an anti-adhesion composition which forms liquid crystals through sol-gel phase transition when exposed to a biological fluid selected from water, saline solution, peritoneal fluid, blood, nutrients or anti-coagulant solution,
Based on total weight,
25-90 wt% of amphipathic lipids of phospholipids;
10 to 75 wt% of oil or fatty acid derived from animal, plant or mineral origin selected from the group consisting of squalene, lanolin, mineral oil, paraffin, petrolatum, almond oil, corn oil, cottonseed oil, olive oil, peanut oil, safflower oil, grapeseed oil, sesame oil, soybean oil, argan oil, canola oil, castor oil, coconut oil, sunflower seed oil, palm oil, avocado oil, medium chain triglycerides, caproic acid, caprylic acid, capric acid, myristoleic acid, palmitoleic acid, oleic acid, linoleic acid, linolenic acid, arachidonic acid, eicosapentaenoic acid, eruic acid and docosahexanoic acid; and
10 to 35 wt% of auxiliary solvent of ethanol or propylene glycol;
An anti-adhesion composition characterized by comprising a mixed solution of . delete delete delete delete delete delete