Patent Application
20230295424
The present invention relates to a composition for forming hydrogel, a hydrogel formed by photo-crosslinking the same, and a method for preparing the hydrogel, and more specifically, a composition which comprises a polyglutamic acid polymer having a photo-crosslinkable functional group introduced therein and a photo-initiator, and is capable of forming a hydrogel in situ through photo-crosslinking, a hydrogel which is formed by photo-crosslinking the composition, and has excellent biocompatibility and adhesive strength in wet environment, flexibility, and ease of use so that it can be utilized as tissue adhesive, surgical sealant, adhesion barrier, etc., and a method for preparing the hydrogel.
Products such as tissue adhesives, surgical sealants and agents for preventing adhesion between tissues are applied to tissues such as dura mater, blood vessels, cornea, lung, intestine, etc. during surgery to help patients recover after surgery through roles of sealing and wound healing, or applied to surgical areas where adhesion between tissues can be formed, and play a role of preventing formation of the adhesion between tissues.
The currently approved tissue adhesives and sealants include cyanoacrylate-based adhesive (Dermabond by Ethicon), fibrin gel utilizing blood coagulation mechanism (Tisseel by Baxter), PEG-based synthetic sealant (Duraseal by Integra lifesciences), etc.
However, in spite of strong adhesion strength thereof, cyanoacrylate-based adhesive has a problem in safety due to cytotoxicity. Fibrin gel has a risk of infection due to the use of ingredient derived from bovine or human source, and it also has low viscosity and mechanical strength, by which in some cases it can neither be applied to the desired area nor perform the function of sealing properly. PEG-based synthetic sealant has good physical properties as a sealant, but has a problem in that it lacks adhesive strength in wet environment and falls off from organs having repetitive movements without being attached thereto. In addition, since the above products require separate procedures of dissolution and assembly before use, usability is low.
Accordingly, there is a demand for the development of new materials that can be suitably used as tissue adhesive, surgical sealant, adhesion barrier, etc., without the above-mentioned disadvantages of the prior art.
The purpose of the present invention is to provide a composition capable of forming a hydrogel which has excellent biocompatibility, safety with low cytotoxicity, excellent adhesion to tissues in wet environment, flexibility, and ease of use in the medical field, and thus can be utilized as tissue adhesive, surgical sealant, adhesion barrier, etc., and a hydrogel formed by photo-crosslinking the composition.
To achieve the above purpose, the present invention provides a composition for forming hydrogel, comprising: a poly(γ-glutamic acid) (PGA) having a photo-crosslinkable functional group introduced therein; and a photo-initiator.
In an embodiment, the composition for forming hydrogel may be a composition for forming hydrogel, comprising: a first component comprising a poly(γ-glutamic acid) (PGA) having a photo-crosslinkable functional group introduced therein; and a second component comprising a photo-initiator.
The present invention also provides a hydrogel formed by photo-crosslinking the above composition for forming hydrogel of the present invention.
The present invention also provides a method of preparing a hydrogel, comprising a step of irradiating the above composition for forming hydrogel of the present invention with ultraviolet ray.
The composition according to the present invention forms a hydrogel in situ after being applied to tissues in the medical field and the hydrogel adheres to the tissues, and thus can be utilized as tissue adhesive, sealant, adhesion barrier, etc., and since properties (burst pressure, swelling ratio, degradation time, flexibility, etc.) thereof are sufficient to be utilized as sealant, it can play a role of sealing while maintaining the properties in the body during recovery of the patient's wound area. In addition, since the gel formation time is as short as 10 seconds (more preferably, 5 seconds), it is cured immediately after application to the tissue surface so as to make sealing of the wound area very easy, and it does not flow down surrounding organs so as to reduce unnecessary use of product, and due to the rapid gel formation, it can provide convenient use and reduced operation time. Furthermore, because of the flexibility and excellent adhesion, it can be used in moving organs such as lung, intestine, etc. Also, the composition and hydrogel according to the present invention can be employed in a medical device such as patch or implant material, etc., and can play a role of attaching the device to tissue, or a role of preventing adhesion between tissues after operation.
The present invention is explained in more detail below.
The composition for forming hydrogel of the present invention comprises a poly(γ-glutamic acid) (PGA) having a photo-crosslinkable functional group introduced therein; and a photo-initiator.
In an embodiment, the photo-crosslinkable functional group may be acrylate group, methacrylate group or a combination thereof.
In an embodiment, the photo-crosslinkable functional group may be derived from acrylate or methacrylate compound which can react with the carboxyl group of poly(γ-glutamic acid) and form a bond.
In an embodiment, such acrylate or methacrylate compound may be one or more selected from glycidyl methacrylate (GMA), 2-hydroxyethyl methacrylate, 2-aminoethyl methacrylate, glycidyl acrylate, 2-hydroxyethyl acrylate, 2-aminoethyl acrylate and salts thereof.
According to an embodiment, by the reaction of the above acrylate or methacrylate compound and poly(γ-glutamic acid), acrylate group or methacrylate group can be introduced as a photo-crosslinkable functional group in the poly(γ-glutamic acid).
In the present invention, “substitution degree of photo-crosslinkable functional group” means, based on 100% of the total moles of carboxyl group in the poly(γ-glutamic acid), the percent (%) of moles of the carboxyl group to which the photo-crosslinkable functional group is bonded.
According to an embodiment, the substitution degree of photo-crosslinkable functional group of the poly(γ-glutamic acid) comprised in the composition for forming hydrogel of the present invention is not especially limited, and for example, it may be 1% or more. If the substitution degree is too less than the above level, the properties of the hydrogel formed from the composition may deteriorate (decrease of burst pressure, increase of swelling ratio, etc.).
More concretely, the substitution degree of photo-crosslinkable functional group of the poly(γ-glutamic acid) may be, for example, 1% or more, 2% or more, 3% or more, 4% or more, 5% or more, 6% or more, 7% or more, 8% or more, 9% or more, 10% or more, 12% or more or 14% or more, and, it may be, for example, 30% or less, 28% or less, 26% or less, 24% or less, 22% or less or 20% or less. The upper limit of the substitution degree of photo-crosslinkable functional group of the poly(γ-glutamic acid) is not especially limited thereto, but if it exceeds the above level, the properties may not be improved further in spite of the increase of the substitution degree.
In an embodiment, the poly(γ-glutamic acid) comprised in the composition for forming hydrogel of the present invention may have a weight average molecular weight of from 10 kDa to 10000 kDa, and more concretely, from 100 kDa to 2000 kDa.
In an embodiment, the photo-initiator may form radical in UV A (320 to 400 nm) region, and preferably it may be a photo-initiator having the maximum absorbance in UV A region. Since the light in UV B (280 to 320 nm) and UV C (100 to 180 nm) regions has a short wavelength and so a high energy, it may cause damage to the tissue and thus application thereof to a living body is inappropriate. Accordingly, in case of irradiation with light in UV A region, use of a photo-initiator having the maximum absorbance in UV B and UV C regions is inappropriate due to its poor radical generation efficiency.
According to an embodiment of the present invention, as a photo-initiator having the maximum absorbance in UV A region, the photo-initiator may be phosphorus-containing aromatic photo-initiator compound, azo-based photo-initiator compound, flavin-based photo-initiator compound, or a combination thereof, but it is not limited thereto.
In an embodiment, the phosphorus-containing aromatic photo-initiator compound may be aromatic phosphinate, aromatic phosphonate, aromatic phosphine oxide, or a combination thereof, and, for example, as that having aromatic group such as benzoyl or terephthaloyl, it may be phosphinate or a salt thereof, phosphonate or a salt thereof, phosphine oxide or a salt thereof, or a combination thereof, but it is not limited thereto.
More concretely, the phosphorus-containing aromatic photo-initiator compound may be lithium phenyl-2,4,6-trimethylbenzoylphosphinate (LAP) (maximum absorbance wavelength: 375 nm) of the following structure, lithium bisacylphosphine oxide (BAPO-OLi) (e.g., lithium phenylbis(2,4,6-trimethylbenzoyl)phosphine oxide) (maximum absorbance wavelength: 375 nm), or a combination thereof, and still more concretely, it may be LAP.
In an embodiment, the azo-based photo-initiator compound may be azo nitrile-based, azo ester-based, azo amide-based, azo amidine-based, or macroazo-based photo-initiator compound, or a combination thereof, but it is not limited thereto.
More concretely, the azo nitrile-based photo-initiator may be V-60, V-65, V-59, V-70, V-40, etc. of Wako Pure chemical Industris. Ltd., the azo ester-based photo-initiator may be V-601, etc. of Wako Pure chemical Industris. Ltd., the azo amide-based photo-initiator may be VA-086, VA-085, VA-080, Vam-110, Vam-111, VF-096, etc. of Wako Pure chemical Industris. Ltd., the azo amidine-based photo-initiator may be V-50, VA-044, VA-046B, Aam-027, VA060, VA-057, VA-061, etc. of Wako Pure chemical Industris. Ltd., and the macroazo-based photo-initiator may be VPS-0501, VPS-1001, VPE-0201, VPE-0401, VPE-0601, VPTG-0301, etc. of Wako Pure chemical Industris. Ltd., but it is not limited thereto.
Still more concretely, the azo-based photo-initiator compound may be an azo amide-based photo-initiator compound, for example, 2,2′-Azobis[2-methyl-N-(2-hydroxyethyl)propionami de] (VA-086) (maximum absorbance wavelength: 365 nm).
In an embodiment, the flavin-based photo-initiator compound may be riboflavin, but it is not limited thereto.
In an embodiment, the concentration of the photo-initiator comprised in the composition for forming hydrogel of the present invention is not especially limited, and it may be a concentration suitable for forming hydrogel by irradiation. For instance, the concentration of the photo-initiator comprised in the composition may be 0.1 mM or higher, 0.5 mM or higher, 1 mM or higher or 1.5 mM or higher, and it may be 10 mM or lower, 7 mM or lower, 5 mM or lower, or 3 mM or lower, but it is not limited thereto.
In an embodiment, when irradiated with ultraviolet ray, preferably irradiated with ultraviolet ray in UV A region (for example, 365 nm wavelength), the composition for forming hydrogel of the present invention may be crosslinked within 10 seconds (more preferably, within 5 seconds) (for example, 1 to 10 seconds, or 1 to 5 seconds) and may form hydrogel. At this time, the wavelength of the ultraviolet ray inducing the photo-crosslinking may be selected properly according to the type of photo-initiator comprised in the composition. For instance, when lithium phenyl-2,4,6-trimethylbenzoylphosphinate (LAP) is used as the photo-initiator, the wavelength of the ultraviolet ray inducing the photo-crosslinking may be, for example, 365 nm.
Thus, other aspects of the present invention provide a hydrogel formed by photo-crosslinking the composition for forming hydrogel of the present invention, and a method of preparing a hydrogel, comprising a step of irradiating the composition for forming hydrogel of the present invention with ultraviolet ray, and more preferably, with ultraviolet ray in UV A region.
In an embodiment, the composition for forming hydrogel of the present invention may be a composition for forming hydrogel, comprising: a first component comprising a poly(γ-glutamic acid) (PGA) having a photo-crosslinkable functional group introduced therein; and a second component comprising a photo-initiator.
Thus, when applied to the tissue surface and then irradiated with ultraviolet ray, the composition for forming hydrogel of the present invention is crosslinked in situ, preferably within 10 seconds (more preferably, within 5 seconds) and forms hydrogel, and the hydrogel can perform tissue adhesion or sealing.
Thus, according to an embodiment, the hydrogel of the present invention can be utilized for use of tissue adhesive, sealant, or adhesion barrier.
The present invention is explained in more detail through the following Examples. However, the following examples are only to illustrate the present invention, and the scope of the present invention is not limited thereby in any manner.
Poly(γ-glutamic acid) (PGA) and glycidyl methacrylate (GMA) were reacted to prepare poly(γ-glutamic acid) having introduced methacrylic (MA) group as a photo-crosslinkable functional group (PGAGMA). The substitution degree of MA in the prepared PGAGMA was controlled by changing the amount of GMA reacting with PGA.
The above-prepared poly(γ-glutamic acid) having introduced methacrylic (MA) group (PGAGMA) and a photo-initiator were dissolved in a buffer to prepare the composition for forming hydrogel.
As the photo-initiator of Examples, lithium phenyl-2,4,6-trimethylbenzoylphosphinate (LAP) and 2,2′-azobis[2-methyl-N-(2-hydroxyethyl)propionamide] (VA-086) were used, whereas as the photo-initiator of Comparative Examples, Irgacure 2959 of the following structure was used.
The prepared composition for forming hydrogel was irradiated with ultraviolet ray (UV) of 365 nm wavelength for photo-crosslinking, and thereby the hydrogel was formed.
For each of the compositions prepared with various types and concentrations of photo-initiator as shown in Table 1 below, the gelation time was measured, and the results are shown in Table 1 below.
As shown in Table 1 above, in case of using LAP and VA-086 as the photo-initiator, the radical generation efficiency was high when irradiated with ultraviolet ray of 365 nm wavelength, i.e., in UV A region (320 to 400 nm), and thus gel formation was possible within short time even with relatively low photo-initiator concentration and low light intensity (UV intensity).
To the contrary, in case of using Irgacure 2959 as the photo-initiator, gel was not formed when the photo-initiator concentration was low, and it was hard to shorten the gelation time within 10 seconds even if the light intensity was increased.
When a gelation time is too long (i.e., 10 seconds or longer), the composition flows down before gelation so that the gel is difficult to apply and adhere to the desired area, and thus it is not suitable for use as a sealant.
Each of the compositions comprising PGAGMA prepared with changing substitution degree of MA and LAP (2 mM) was irradiated with ultraviolet ray of 365 nm wavelength to form hydrogel.
Then, the initial weight (W0) of the prepared hydrogel was measured and the sample was immersed in PBS, and then stored in a shaking incubator at 37° C. for 24 hours. Then, the excess water on the sample surface was sufficiently wiped off, the weight of the sample (W1) was measured, and the swelling ratio was calculated through the following equation, and the results are shown in
Swelling ratio (%)=((W1−W0)/W0)*100%
As shown in
Each of the compositions comprising PGAGMA prepared with changing MA substitution degree and LAP (2 mM) was irradiated with ultraviolet ray of 365 nm wavelength to form hydrogel.
The following test was conducted according to ASTM2392-04 (Standard Test Method for Burst Strength of Surgical Sealants), and the results are shown in
The collagen casing was washed sequentially with distilled water and ethanol, and the washed collagen casing was punched to make a 3 mm hole therein, and then a teflon ring (inner diameter: 15 mm, thickness: 2 mm) was placed on the center of the hole.
Then, each of the compositions comprising PGAGMA prepared with changing substitution degree of MA and LAP (2 mM) was irradiated with ultraviolet ray of 365 nm wavelength to form hydrogel.
Then, the hydrogel-attached collagen casing was mounted on an instrument for measuring burst pressure, and whiled flowing PBS at a rate of 2 mL/min, applied maximum hydraulic pressure was measured.
As shown in
While bending the hydrogel sample prepared according to the present invention by hand, photos thereof were taken as shown in
As shown in
| Number | Date | Country | Kind |
|---|---|---|---|
| 10-2020-0098415 | Aug 2020 | KR | national |
| Filing Document | Filing Date | Country | Kind |
|---|---|---|---|
| PCT/KR2021/010330 | 8/5/2021 | WO |
