KR20070018099A - Agent for visualizing transparent tissue - Google Patents

Abstract

Disclosed is a means for enhancing the visibility of the transparent tissues of the eye such as the vitreous, the lens or the cornea and improving safety. Said means can not completely dissolve fine particles made from a polymer compound, in particular 1 g thereof, in less than 30 mL of water at 20 ° C. within 30 minutes, and fine particles having an average particle diameter of 0.01 to 500 μm, particularly biodegradable among them. Microparticles made of a high molecular compound and / or a water-soluble high molecular compound.

Description

Transparent tissue visualizer {AGENT FOR VISUALIZING TRANSPARENT TISSUE}

The present invention is a transparent tissue visualizing agent for improving the visibility of the transparent tissue of the eye at the time of surgery by injecting or spreading the transparent tissue, and a method of visualizing the transparent tissue of the eye by the application of the agent to improve the visibility It is about.

The eye is composed of transparent tissues, many of which are corneal, lens and vitreous in order to collect light from the outside world and project it onto the optic cells. The vitreous body is in contact with the retina, and many retinal diseases, including diabetic retinopathy, serve as a bridgehead for proliferative tissue generated in the retina. Tissues that proliferate in the vitreous form fibers, which retract the retina, causing retinal detachment, which can lead to blindness if left untreated. Thus, vitreous bodies containing such proliferating tissues are often completely removed by surgical surgery.

In vitreous debridement surgery, it is necessary to completely remove not only the proliferative tissues adhered to the retina, but also the vitreous that is the bridgehead of the tissue proliferation as completely as possible. The operation is performed while flowing intraocular perfusion liquid for surgical operation into the eyeball. However, the vitreous is a transparent structure and there is little difference in refractive index with the intraocular perfusion liquid. For this reason, in the area | region through which the surgical microscope passed through without a countermeasure, transparent tissues lack visibility, making it difficult to discriminate the position of the said tissue, and its complete removal is not easy. As a solution, during a vitreous removal procedure, a steroid suspending agent, such as a triamcinolone preparation (Kenacort-A®), is injected into the vitreous cavity (a hollow portion that could be aspirated off the vitreous center) to disperse the suspension and adhere to the vitreous Visualization of the vitreous was carried out (see Non-Patent Document 1). This method increases the visibility of the vitreous in the surgical area, facilitates the surgery, and enables complete removal of the vitreous. However, with respect to the application of the steroid to the vitreous, elevated intraocular pressure and cataract progression as side effects have been reported (see Non-Patent Documents 2 and 3). Therefore, the use of a steroidal suspending agent for the purpose of improving the visibility in vitreous removal surgery has the possibility of developing the same side effects.

On the other hand, the method of dyeing and visualizing the proliferative film | membrane or retinal epithelium accompanying proliferative vitreoretinopathy (PVR) with the aqueous solution which dissolve | stained the pigment | dye like trypan blue is also reported (refer patent document 1). However, since the use of the pigment darkens the surgical area and the vitreous itself cannot be emphasized in the surgical area, the improvement of visibility according to this is very insufficient.

For example, in cataract surgery, after removal of the nucleus and cortex in the capsular bag, intraocular lenses are inserted, but the remaining cortex, which is not removed near the posterior capsule, proliferates and becomes cloudy over time after surgery, and becomes late. It is known to cause cataracts. Therefore, even in cataract surgery, a means for visualizing the transparent cortical portion and facilitating complete removal is necessary.

[Patent Document 1] WO 99/058159

[Non-Patent Document 1] Sakamoto, T., et al., Graefe's Archive for Clinical and Experimental Ophthalmology, 240: p.423 (2002).

[Non-Patent Document 2] Challa, J.K. Et al., Australian and New Zealand Journal of Ophthalmology, 26: p. 277 (1998)

[Non-Patent Document 3] Wingate, R.J. Et al., Australian and New Zealand Journal of Ophthalmology, 27: p.431 (1999)

Disclosure of the Invention

Problems to be Solved by the Invention

Against this background, it is an object of the present invention to provide a means for improving the visibility of vitreous, crystalline or corneal surgery of the transparent tissues of the eye, which is easy to use and excellent in safety while achieving sufficient visibility. It aims to do it.

Means to solve the problem

As a result of the examination of the above problems, the inventors of the present invention have a pharmacological action as a polymer compound, and a polymer compound which is gradually dissolved in body fluids and is discharged and / or absorbed by the turnover of the intraocular liquid, particularly preferably biodegradable. Injecting or dispersing the polymer compound in the form of microparticles into the transparent tissue of the eye, while it is present as microparticles, can attach to the surface of the transparent tissue and scatter visible light into the surgical area, thus achieving excellent visibility without causing safety concerns. I found something that could be. The present invention has been completed through further studies based on this finding. That is, this invention provides the following.

(1) A transparent tissue visualizing agent for enhancing the visibility by contacting transparent tissue of an eye containing fine particles made of a high molecular compound.

(2) The transparent tissue visualization agent according to (1), wherein 1 g of the fine particles cannot be completely dissolved in less than 30 mL of water at 20 ° C within 30 minutes.

(3) The transparent tissue visualizing agent according to (1) or (2), wherein the average particle diameter of the fine particles is 0.01 to 500 µm.

(4) The transparent tissue visualizing agent according to any one of (1) to (3), wherein the polymer compound is a biodegradable polymer compound and / or a water-soluble polymer compound.

(5) The transparent tissue visualizing agent according to (4), wherein the biodegradable polymer compound is selected from fatty acid polyesters, polysaccharides and derivatives thereof.

(6) The transparent tissue visualizing agent according to (4), wherein the water-soluble high molecular compound is selected from an acrylic polymer compound and a styrene polymer compound.

(7) The polymer compound according to any one of (1) to (3), wherein the polymer compound is starch, soluble starch, pregelatinized starch, cellulose, acetyl cellulose, hydroxypropylmethyl cellulose, chitosan, chitin, dextran, poly Lactic acid, polyglycolic acid, lactic acid-glycolic acid copolymer, polyhydroxybutyric acid, polyhydroxy valeric acid, polycaprolactone, hydroxybutyric acid-glycolic acid copolymer, lactic acid-caprolactone copolymer, polyethylene succinate and polybutyl A transparent tissue visualizing agent selected from the group consisting of lene succinate, sodium polyacrylate, sodium methacrylate and sodium polystyrene sulfonate.

(8) The transparent tissue visualizing agent according to (7), wherein the polymer compound has an average molecular weight of 500 to 200000.

(9) The transparent tissue visualizing agent according to (8), further comprising a polyvinyl compound and / or a polyol.

(10) The transparent tissue visualizing agent according to (9), which contains 0.05 to 10 parts by weight of the polyvinyl compound and / or 0.05 to 10 parts by weight of the polyol based on 1 part by weight of the fine particles.

(11) The transparent tissue visualizing agent according to (9) or (10), wherein the polyvinyl compound is polyvinyl pyrrolidone and / or polyvinyl alcohol.

(12) The transparent tissue visualizing agent according to (9) or (10), wherein the polyol is mannitol.

(13) The transparent tissue visualizing agent according to any one of (1) to (12), further comprising an aqueous medium, wherein the fine particles are dispersed in the aqueous medium.

(14) The transparent tissue visualization agent according to (13), wherein the content of the fine particles in the transparent tissue visualization agent is 0.005 to 10 w / v%.

(15) The transparent tissue visualizing agent according to (13) or (14), further comprising a polyvinyl compound and / or a polyol.

(16) The transparent tissue visualizing agent according to (15), which contains a polyvinyl compound at a concentration of 0.1 to 5 w / v% and / or a polyol at a concentration of 0.1 to 5 w / v%.

(17) The transparent tissue visualizing agent of any one of (1) to (16) above, which is an injection preparation.

(18) A transparent structure comprising a seal in a single mixing and discharging means, enclosed in a state in which the powder composition comprising the fine particles according to any one of (1) to (12) and the aqueous medium are kept in contact with each other. Visualizing agent.

(19) A method for improving the visibility of transparent tissue of an eye, comprising contacting the composition comprising any one of the above (1) to (17) with the transparent tissue of the eye.

(20) Use of the microparticles defined in any one of (2) to (8) above for the production of a transparent tissue visualizing agent that contacts and visualizes the transparent tissue of the eye.

Effects of the Invention

The present invention as defined above significantly enhances the visibility by contacting the transparent tissue of the eye, ie, the vitreous, the lens, the cornea, with the transparent tissue that is difficult to see in the surgical area. To allow for easier handling and thus to achieve the surgical purpose consistently and easily. Moreover, since microparticles | fine-particles which do not have a pharmacological action are used, it does not cause unnecessary drug response or side effects to a living body. In particular, the use of soluble and / or biodegradable particulates dissolves and / or decomposes over a period of time, even if some of the particulates remain after surgery, and then discharged or absorbed and disappeared from the eye tissue. It does not cause a problem.

Brief description of the drawings

1

Schematic cross section showing one embodiment of a double chamber shaped syringe filled clear tissue visualization agent

Explanation of the sign

(1) = double chamber type syringe, (2) = movable separator, (3) = solid phase, (4) = liquid bed side, (7) = discharge flow path, (8) = piston, (9) = peripheral flow path

To practice the invention  Best form for

In the ophthalmic surgery, the transparent tissue visualizing agent in the present invention improves the visibility of the transparent tissue by injecting or scattering the transparent tissue of the eye. In particular in intraocular surgery involving vitreous removal, such as diabetic retinopathy, retinal vein occlusion, macular edema, diabetic macular disease, macular hole, retinal epithelial disease, and thermoparking retinal detachment, the vitreous cavity of the present invention When a transparent tissue visualizing agent is injected (in the form of a potion) or dispersed (in the form of a powder), a myriad of fine particles adhere to the vitreous gel and light scatters along the surface of the vitreous gel when irradiated with visible light. Therefore, the visibility of the vitreous body is improved, and the eye can be easily confirmed in the surgical region through, for example, a surgical microscope. In addition, in surgical operations involving the removal of the nucleus and cortex of the lens, infusion or scattering in the capsular bag causes numerous microparticles to adhere to the nucleus and cortex of the lens, causing light scattering along its surface. The visibility also improves as well.

In the present invention, as the "polymer compound", a macromolecular compound pharmacologically inactive to the tissue and the body of a mammal, in particular, the human eye can be used. Here, the polymer compound used in the present invention is a solid at room temperature, and there is no particular limitation on its molecular weight as long as it can be molded into particles. Usually, the molecular weight is 500 or more, preferably 1000 or more, more preferably 1500 or more, and particularly preferably 2000 or more. It is usually 200000 or less, preferably 150000 or less, more preferably 100000 or less, particularly preferably 50000 or less. However, as long as the object of the present invention can be achieved, these ranges may be exceeded.

As a high molecular compound, various things can be used, but it is preferable that 1 g of microparticles | fine-particles formed with a given polymer do not melt | dissolve completely in water less than 30 mL at 20 degreeC within 30 minutes, and it is such a solubility Is sufficient to make the particulates resistant to overall dissolution and remain as light scattering sources during surgical operations. This degree of solubility corresponds to solubility ranging from "slightly insoluble" to "hard to melt", "very hard to melt" and "almost insoluble" as defined in the 14th revised Japanese Pharmacy. In the pharmacy room, solubility is defined based on the degree of melting within 30 minutes when 1 g of the solute is added to the solvent and shaken vigorously for 30 seconds at 20 ± 5 ° C. for 5 minutes. In this invention, based on the same definition, when 1 g of microparticles | fine-particles are put in water at 20 degreeC, and fully stirred for 30 minutes, microparticles | fine-particles whose solubility corresponds to any of the following are used preferably.

(1) slightly insoluble (ie, completely insoluble in water less than 30 mL, but completely soluble in water in a range of 30 mL or more and less than 100 mL),

(2) difficult to dissolve (ie, completely insoluble in water less than 100 mL, but completely soluble in water in a range of 100 mL or more and less than 1000 mL)

(3) very difficult to dissolve (ie, cannot be completely dissolved in less than 100 mL of water, but completely soluble in a range of 1000 mL or more and less than 10000 mL)

(4) Almost insoluble (ie, at least 10000 mL or more of water is required for complete dissolution).

In addition, when the high molecular compound takes the form of a salt, sodium salts, potassium salts, hydrochlorides and the like are given as preferred examples, but not limited thereto.

There is no obvious limitation on the average particle diameter of the fine particles composed of the high molecular compound. However, in consideration of ease of handling at the time of use or scattering of visible light, it is usually preferably 0.01 to 500 µm, more preferably 0.1 to 200 µm, still more preferably 1 to 60 µm.

Considering the possibility that some of the fine particles of the polymer compound remain in the tissue after the surgery, it is preferable that the polymer compound is biodegradable or somewhat water-soluble, and even if there are residual particles in the eye tissue after the surgical operation, it is decomposed over time or degraded. If not, they are released from, or absorbed by, or re-dissolved from the eye tissue with the turnover of the liquid present in the tissue of the eye in which they are dissolved and lost from the eye tissue.

Various biodegradable high molecular compounds can be used in this invention. Examples include polylactic acid, polyglycolic acid, lactic acid-glycolic acid copolymer, polyhydroxybutyric acid, polyhydroxy valeric acid, polycaprolactone, hydroxybutyric acid-glycolic acid copolymer, lactic acid-caprolactone copolymer, polyethylene succinate Fatty acid polyesters such as polybutylene succinate, and the like; Examples of the polysaccharide and its derivatives include starch derivatives such as starch, soluble starch and pregelatinized starch, cellulose and cellulose derivatives such as acetyl cellulose and hydroxypropyl methylcellulose, as well as chitosan, chitin and dextran, but are not limited thereto. It doesn't work.

With respect to the polymer compound which can be used in the present invention, there is no specific limitation on the degree of water solubility. For example, it is sufficient if there is solubility that it dissolves when stirred for one week at 37 degreeC with excess water. Even after the surgical operation, the fine particles may remain in the guide, the amount is usually very small, and there is no problem if it gradually dissolves and loses its form as a solid. Examples of such polymer compounds other than biodegradable polymer compounds include, but are not limited to, acrylic polymers and styrene polymers, such as sodium polyacrylate, sodium methacrylate, sodium polystyrenesulfonate, and the like.

The transparent tissue visualizing agent of the present invention may include one or both of a polyvinyl-based compound and a polyol together with the fine particles made of a high molecular compound. The polyvinyl compound and the polyol have a preferable function of increasing the dispersibility of the fine particles composed of the high molecular compound. Examples of particularly preferred polyvinyl compounds include, but are not limited to, polyvinylpyrrolidone and polyvinyl alcohol, and examples of particularly preferred polyols are mannitol. Thus, water-soluble, inert polyvinyl-based compounds and polyols having no pharmacological action can be used as desired. The amount of the polyvinyl compound and the polyol may be about 0.05 to 10 parts by weight based on 1 part by weight of the fine particles of the polymer compound. However, since there is some effect according to at least the amount than this range, and even more, there is no particular problem, it can be out of the range.

The transparent tissue visualizing agent of the present invention may be in powder form or may be in a form in which fine particles made of a high molecular compound are dispersed in an aqueous medium (suspension agent). When the said formulation is in powder form, it may be disperse | distributed to the transparent tissue of an eye as it is, and may be mixed and injected with an intraocular perfusion liquid etc. before use. Typically, intraocular perfusion-cleaning fluids (e.g., Opeguard MA®, Operguard neo kit®, etc., available from Senju Pharmaceutical Co., Ltd.) or artificial tears are suspended in a conventional manner. Can be used after. When provided in the form of a suspending agent, it may be poured directly into the transparent tissue, directly applied, or applied via a needle.

An "aqueous medium" used to provide the transparent tissue visualizing agent of the present invention in the form of a suspending agent is acceptable for tissues of the eye, in particular intraocular tissues, such as isotonic agents, buffers and the like, such as salts or sugars, if desired Possible media are one or more additives and water. Examples thereof include such intraocular perfusion-rinse liquids, including but not limited to Opeguard MA (registered by Senju Pharmaceutical Co., Ltd.), which are distilled water for injection, buffered saline, commercial intraocular perfusion solution or equivalent thereof. Trademarks), Operguard neo kit (registered trademark, etc.), or artificial tears or their equivalents.

When the transparent tissue visualizing agent of the present invention is in the form of a suspending agent, in order to achieve a stable visibility improvement effect, the content of the microparticles made of a high molecular compound in the suspending agent is preferably in the range of 0.005 to 10 w / v%, 0.01 to 5 It is more preferable to set it as w / v%, and it is especially preferable to set it as 0.1-2 w / v%. However, it is possible to obtain visibility even at a lower content than this, for example, 0.0001 w / v%, and even higher contents than this can be used without any inconvenience in handling such as dispersion of fine particles. Can be. For convenience, the content can usually be appropriately determined in the range of 0.01 to 5 w / v%. In addition, in the case of further containing at least one of the above-described polyvinyl compounds and polyols in order to increase the dispersibility of the fine particles, the concentration of any of these can be appropriately set in the range of 0.1 to 5 w / v%. However, if the concentration is less than this range, there is a slight effect according to the amount, and if more, there is no particular problem, so it can be out of this range.

The transparent tissue visualizing agent of the present invention can be provided as a powder or a suspending agent in which fine particles are dispersed in an aqueous medium as described above. Furthermore, a powder consisting of microparticles of a high molecular compound (and, optionally, one or more additives such as polyvinyl compounds, polyols) and an aqueous medium used to disperse the powder prior to use in a surgical operation to prepare a suspending agent, It may be provided in the form of being enclosed in a single mixing and discharging means in a state of being separated from contact. In this case, the aqueous medium may be one of those described above.

In the present invention, the "mixing and discharging means" means that the powder and aqueous medium which are separated and encapsulated can be mixed by operation from the outside, and the formed mixture can be discharged to the outside by operation from the outside. no limits. Accordingly, syringes in the form of various double chambers are well known. Double chamber syringes are typically cylindrical with a needle at one end (or already equipped with a needle) and with a discharge flow path, and at the other end with a piston that prevents liquid from passing through. The separator is densely inserted and the separator which is movable in the front-rear direction is inserted in the middle part of the cylinder so that liquid cannot pass, and forms two chambers of the front and the rear in a cylinder. In front of the separator, an elongated peripheral flow path extending in the longitudinal direction longer than the thickness of the separator is provided by forming a recess in the inner surface of the cylinder. The front chamber is usually filled with a dry composition, for example, in powder form, and the liquid chamber (for example, a buffer) which is a medium for mixing with the dry composition is enclosed in the rear chamber. Mixing and discharging are performed as follows. First, the piston inserted in the rear end is advanced, and the movable separator is advanced in the barrel under water pressure generated thereby to reach near the center of the peripheral flow path. Continue advancing the piston until it pushes the liquid composition of the rear chamber through the surrounding flow channel into the front chamber, mixing the contents in the front chamber, and then pressing the piston further to advance the movable separator to discharge the mixture from the discharge flow path. Let's do it. It is particularly advantageous to provide the transparent tissue visualizing agent of the present invention in a form enclosed in such a single mixing and dispensing means in a state where the components are not in contact with each other, greatly increasing the convenience of use in a surgical operation.

The transparent tissue visualizing agent of the present invention is an additive acceptable as a medicament, for example, isotonic agents (salts such as sodium chloride and potassium chloride; sugars such as glycerol and glucose; polyol types such as sorbitol, mannitol and propylene glycol; boric acid and borate) Etc.), buffers (phosphate buffer, acetic acid buffer, boric acid buffer, carbonic acid buffer, citric acid buffer, tris buffer, etc.), thickeners (hydroxyethylcellulose, hydroxypropylcellulose, hydroxypropylmethylcellulose, carboxycellulose sodium, polyvinyl alcohol , Polyvinylpyrrolidone, polyethylene glycol, sodium alginate, etc.), stabilizers (sodium bisulfite, ascorbic acid, sodium ascorbate, dibutylhydroxytoluene, etc.), pH adjusters (hydrochloric acid, sodium hydroxide, phosphoric acid, acetic acid, etc.) Etc. can be contained as desired. In addition, it is possible to contain the pharmaceutically active ingredient in the transparent tissue visualization agent of the present invention, unless it is contrary to the object of the present invention. When the transparent tissue visualizing agent of the present invention is in the form of a suspending agent (or in a form in which the powder and the aqueous medium are separated and enclosed), the pH of the (or after mixing) is usually 4.0 to 8.0, preferably Adjusted to range from about 5.0 to 7.5.

The present invention also provides a method for improving the visibility of transparent tissues of the eye, comprising contacting the composition comprising the microparticles with the transparent tissue of the eye, and transparent tissue visualization designed to contact and visualize the transparent tissue of the eye. Also provided is the use of the microparticles described above for the preparation of a medicament.

Hereinafter, the present invention will be described in more detail with reference to Examples, Comparative Examples, and Test Examples. However, the present invention is not intended to be limited to these examples.

Test Example 1 Vitreous Visibility Enhancement Test

After euthanizing the colored rabbits, the eyeballs (6 from 3) were taken out. The cornea was excised along the corneal ring with scissors. The iris and the lens were removed, and after leaving the optic cup, the cross was cut from the anterior eye side and opened. The vitreous that began to flow to the outside was removed, and the retina was exposed with the vitreous body partially attached. An aqueous formulation (suspension) containing 1 w / v% soluble starch was prepared according to the following formulation, and about 100 μl thereof was added dropwise to the retinal portion. The preparation dropping site was observed using an optical microscope. As a result, in all six eyes, soluble starch microparticles | fine-particles adhered to the vitreous body, and the visibility of the vitreous body was improved by it.

(Formulation)

Soluble starch ... 1.0 g

Opeguard MA ... to 100 mL total

(For Opeguard MA, see Test Substances and Reagents section below.)

[Test Example 2] Confirmation of visibility using the extraction money

1. Organization used:

We used money purchased from meat processing plant. After removing the pericular tissue immediately after purchase, only the isolated eye was immersed in a low molecular weight Dextran L Injection (Otsuka Pharmaceutical Co., Ltd.) and preserved until the start of the test.

2.Test substance and reagents:

The following were used as test substance and reagent.

(1) Soluble Starch (manufactured by Nacalai Tesque, Inc., Reagent Code 33131-42, Specification GR (Guaranteed Reagent))

(2) chitosan (manufactured by Funakoshi Co., Ltd., reagent code K-03, particle size: 30 mesh pass: 95% or more)

(3) PLA-0005 (manufactured by Wako Pure Chemical Industries, Ltd., D, L-polylactic acid (molecular weight 5000))

(4) PLA-0010 (manufactured by Wako Pure Chemical Industries, Ltd., D, L-polylactic acid (molecular weight 10000))

(5) PLGA-5005 (manufactured by Wako Pure Chemical Industries, Ltd., lactic acid-glycolic acid copolymer (molecular weight 5000))

(6) Povidone K-30 (manufactured by BASF Japan Ltd., Japan Pharmacy)

(7) Goshenol EG-05 (manufactured by Nippon Synthetic Chemical Industries, Co., Ltd., polyvinyl alcohol EG-05)

(8) D-mannitol (manufactured by Nacalai Tesque, Inc.)

(9) Opeguard MA (manufactured by Senju Pharmaceutical Co., Ltd., intraocular perfusate, containing the following components (per mL): 1.5 mg glucose, 6.6 mg sodium chloride, 0.36 mg potassium chloride, 0.18 mg calcium chloride, 0.3 mg magnesium sulfate Sodium bicarbonate 2.1 mg)

3.Method of preparing a transparent tissue visualizing agent:

3-1. Preparation of the transparent tissue visualizing agent of Comparative Example 1 and Examples 1 to 5:

See Table 1. PLA 0005 was slowly thrown into an A-O jet mill (Seishin Enterprise Co., Ltd.) and pulverized (pressurization conditions: grinding nozzle 0.4 MPa, pusher nozzle 0.4 MPa). Opeguard MA was added to the obtained powder to prepare a 2% and 1% suspension to prepare Examples 5 and 4, respectively. Example 4 The suspension was diluted 10-fold with Opeguard MA to give a suspension of Example 3. Example 3 The suspension was diluted 10-fold with Opeguard MA to give a suspension of Example 2. Example 2 The suspension was diluted 10-fold with Opeguard MA to give a Comparative Example 1 suspension. 10 mL of the 1% suspension of Example 4 was centrifuged (1000 rpm, 5 minutes) to precipitate large particles, and the supernatant was collected to give Example 1 suspension. 1 mL of the supernatant was collected, and the content ratio of PLA0005 fine particles in the supernatant was determined based on the weight of the dried product (subtracting the solid content originally included in Opeguard MA in the above calculation).

3-2. Preparation of the transparent tissue visualizing agents of Comparative Examples 3 to 5 and Examples 9 to 25:

See Tables 2-4. To purified water, one of the following reagents was added to obtain the indicated concentration, which was filtered through a 0.22 μm hydrophilic filter to give a liquid A for the corresponding example: 1.11% povidone K-30 (Examples 11-13 and Examples) 22 to 25), D-mannitol 1.11% (Example 17 and Examples 22 to 25) and Gosenol EG-05 0.22% (Example 25, when prepared). A 10% PLA0005 (acetone / ethanol = 4/6) solution was prepared, filtered through a 0.22 μm hydrophobic filter to a B liquid. Opeguard MA was used as C liquid.

Each A liquid corresponding to each Example and B liquid which is a polylactic acid solution were mixed at the ratio of 9: 1 as follows. That is, while stirring each A liquid at 700-800 rpm with the stirrer, B liquid was dripped at the speed | rate of about 100 microliters / second, and PLA0005 microparticles | fine-particles were deposited. After stirring this for about 30 minutes, the aggregated mass was removed by the sieve (diameter 106 micrometers), and suspension D was obtained. Lyophilization was carried out to obtain a powder sample E. The powder samples E were dispersed in the C liquid so that the final content of the fine particles was 1 w / v%, thereby being examples 11, 17 and 22. The composition of Example 11 with a particulate content of 1 w / v% was sequentially diluted with Opeguard MA 10 times at a time until the content became 0.001%, whereby the composition with a content of 0.1 w / v% was prepared in Example 10, A composition having a content of 0.01% w / v% was used as Example 9, and a composition having a content of 0.001 w / v% was used as Comparative Example 3. The composition of Example 17 was likewise diluted 10-fold at a time with Opeguard MA, so that the composition with a content of 0.1 w / v% as Example 16 and the composition with a content of 0.01 w / v% as Example 15 were 0.001 The composition of w / v% was made into Example 14, and the composition of content 0.0001 w / v% was made into Comparative Example 4. In addition, the composition of Example 22 was similarly diluted 10 times at a time with Opeguard MA, so that the composition having a content of 0.1 w / v% as Example 21 and the composition with a content of 0.01 w / v% as Example 20 were added. A composition having a content of 0.001 w / v% was used as Example 19, a composition having a content of 0.0001 w / v% was used as Example 18, and a composition having a content of 0.00001 w / v% was set as Comparative Example 5. In addition, the powder sample E used in Example 11 was disperse | distributed to C liquid, and the suspension prepared so that PLA0005 might reach final content 2w / v% and 5w / v% was set as Example 12 and Example 13, respectively. In addition, the suspensions prepared in Example 22 and Example 24 were prepared by dispersing the powder sample E used in Example 22 in a liquid C so that PLA0005 reached final concentrations of 2 w / v% and 5 w / v%. The suspension prepared in Example 25 was prepared by dispersing the powder sample E precipitated in an aqueous solution containing goceol EC-05 in a liquid C so that the PLA0005 reached a final concentration of 1 w / v%.

Lyophilization for the preparation of the compositions of Examples 11-13, 17, and 22-25 was carried out as follows: Suspension D was stored at -40 ° C for 6 hours to freeze, and 100 µm at -40 ° C. The pressure was reduced to Hg or less and dried for 24 hours or more. The temperature was raised by 10 ° C every hour at -40 ° C and continued until + 20 ° C. Furthermore, it dried at +20 degreeC and 100 micrometers Hg or less for 24 hours or more, and set it as the powder sample E.

3-3. Preparation of the transparent tissue visualizing agent of Comparative Example 2 and Examples 6 to 8:

The 100 mL of Example 11 suspension prepared in the same manner as in the above 3-2 was centrifuged (centrifugal condition: 1000 rpm, 5 minutes) to precipitate large particles, and the supernatant was collected. Supernatant was used as the composition of Example 8. In addition, the composition of Example 8 was diluted 10-fold with Opeguard MA to obtain the composition of Example 7, which was diluted 10-fold with Opeguard MA to obtain the composition of Example 6, and diluted 10-fold with Opeguard MA, and Comparative Example 2 It was made into the composition. Content of PLA0005 in supernatant was extract | collected 1 mL of the supernatant of the composition of Example 8, and was calculated from the weight which dried this. In the calculation, the solids content included in povidone and Opeguard MA was subtracted.

3-4. Preparation of the transparent tissue visualizing agent of Comparative Example 6 and Examples 26 to 31:

Referring to Table 5, the soluble starch was gradually added to an A-O jet mill (Seishin Enterprise Co., Ltd.) and ground (pressure conditions: grinding nozzle 0.4 MPa, pusher nozzle 0.4 MPa). Opeguard MA was added to the sample to prepare a suspension having a particulate content of 10 w / v%, to give a suspension of Example 31. The suspension of Example 31 was diluted with Opeguard MA to give a suspension of content of 1 w / v% to Example 30 and the suspension of content of 0.1 w / v% to Example 29 and a suspension of content of 0.01 w / v% In Example 28, the suspension of content 0.005 w / v% was used as Example 27, the content of the 0.0025 w / v% suspension was determined in Example 26, and the content of 0.001 w / v% was determined as Comparative Example 6.

3-5. Preparation of the Clear Tissue Visualizing Agents of Examples 32-35:

Referring to Table 6, PLGA5005 or chitosan was gradually added to an A-O jet mill (Seishin Enterprise Co., Ltd.) and pulverized (pressure conditions: grinding nozzle 0.4 MPa, pusher nozzle 0.4 MPa). Opeguard MA was added to these, and it prepared so that it might become a 1 w / v% suspension, respectively, and the PLGA5005 containing suspension was used in Example 33, and the chitosan containing suspension was carried out in Example 35. In addition, 1 w / v% PLA0010 suspension prepared by adding Opeguard MA was used as Example 34. The suspension of Example 33 was centrifuged (centrifugal conditions: 1000 rpm, 5 minutes) to precipitate large particles, and the supernatant was collected. This supernatant was referred to as Example 32. The PLGA5005 content in the supernatant was determined from the dried weight of 1 mL of the supernatant. In the calculation, the solid content included in Opeguard MA was subtracted.

4. Test Method:

Laser diffraction particle size distribution measuring device (SALD-) was used to determine the particle diameters of the fine particles prepared in Examples 1, 4, 8, 11, 17, 22, 30 and 35. 2100, Shimadzu Corporation) was used to measure the average particle diameter. The average particle diameter of the said microparticles | fine-particles was made into the average value computed by the method shown by Formula (1). That is, in each particle diameter section (x _j , x _{j +1} ) provided by dividing a number (n) over the entire distribution range of the particle diameter, the particle diameter is represented by the logarithm average (as logarithmic) of the particle diameter at both ends of the section. Multiply the volume difference percentage (volume distribution frequency) for the total distribution range of all particles measured in each interval and add the values obtained for each interval over the entire distribution range, and in the mean, from the average value (μ) obtained, The average particle diameter (10 ^μ ) was calculated. In the case where the dispersion of the fine particles due to shaking was difficult because of aggregation, the measurement was performed after dispersion by ultrasonic waves. In addition, since the comparative example and the Example except the above used powder of the same lot as the said sample, particle size measurement was not performed.

(Note 1) Calculation method of average particle size:

[Equation 1]

Where x _j : particle size (μm)

q _j : volume difference% (volume frequency distribution)

For vitreous surgery, a vitreous surgical device (Ocutome®, ALCON) and an ophthalmic surgical microscope (CARL ZEISS) were used. A sclera was cut in parallel with the corneal ring using an ophthalmic surgical knife (20GV-Lance ^™ , ALCON) from a site about 3 mm outside the corneal ring in the donor to prepare a port for perfusion cannula. A perfusion cannula was inserted onto the manufactured pot, and physiological saline (Otsuka Pharmaceurical CO. Ltd.) was constantly flowed during the surgical operation. By using a surgical knife to about 120˚ ophthalmic from the guidance perfusion ports of the outer region about 3 mm around the periphery of the cornea from a cornea ryunbu (20GV-Lance ^TM, ALCON) incision in the cornea in parallel with ryunbu cornea, vitreous cutter I made a port for a vitreous cutter. The vitreous cutter was inserted through the prepared port, the lens and other surrounding tissues were excised, and after confirming that the fundus could be observed under a microscope, the central portion of the vitreous was removed. Using a disposable syringe (5 mL, Terumo Corporation) equipped with a non-bevel needle (22G, Terumo Corporation), 1 mL of transparent tissue visualizing agent was injected into the lumen formed by removal of the central portion of the vitreous from the pot for vitreous cutter. After removing the fine particles suspended in the liquid with a vitreous cutter, the light guide is inserted into the port for the vitreous cutter, and the visibility of the vitreous body due to the adhesion of the fine particles is based on the following evaluation criteria. It evaluated by looking at each site | part of the equatorial part glass body and a rear part glass body (retina side). Regarding the viability and degree of visualization of the vitreous with the transparent tissue visualizing agent, it was determined that the score 2 or 3 of the evaluation table was obtained at one or more of these three sites, and had a viable visualizing effect.

<Evaluation Criteria>

0 ... Adherence of microparticles | fine-particles to a residual glass body is not observed, and the shape of a residual glass body cannot be visually recognized.

1 ... Adherence of microparticles | fine-particles to a residual glass body is observed, but it cannot visually recognize until the shape of a residual glass body.

The shape of the remaining vitreous body can be visually recognized by the adhesion of the fine particles, and the retina is clearly seen through the group of adhered fine particles.

The shape of the remaining vitreous can be visually recognized by the adhesion of the fine particles, and the retina is only blurred through the group of adherent fine particles.

5. Test result:

5-1. Visibility of the Clear Tissue Visualizing Agent Containing PLA-0005:

Table 1-4 shows the result about the visibility improvement effect of the PLA-0005 containing transparent tissue visualizing agent. In order to prepare PLA0005 of different particle diameters, in Comparative Example 1 and Examples 1-5, PLA0005 was grind | pulverized using the A-O jet mill. Moreover, about Comparative Examples 2-5 and Examples 6-25, after melt | dissolving PLA0005 in the organic solvent, microparticles were prepared by making it precipitate as microparticles | fine-particles. In addition, povidone solutions for Comparative Examples 2, 3 and 6 to 13, mannitol solutions for Comparative Examples 4 and 14 to 17, povidone and Comparative Examples 5 and 18 to 24 About mannitol solution and Example 25, PLA0005 was precipitated as fine particles in gosenol and mannitol solution, respectively. Moreover, about Comparative Example 2, Example 1, and Examples 6-8, the high concentration suspension of PLA0005 was centrifuged, supernatant was collected, and finer fine particles were obtained. As a result, microparticles | fine-particles of the range of an average particle diameter of 0.8 micrometer-60.3 micrometers were obtained. As a result of determining the concentration useful as the vitreous visualizing agent of these fine particles by the visibility test, it was PLA0005 fine particles having an average particle diameter of 17.9 µm, and was useful up to a low content of 0.001 w / v%. (Example 18). PLA0005 particles with a minimum particle size of 0.8 μm in the sample were useful up to a low content of 0.003 w / v% (Example 6). PLA0005 particles having a maximum particle size of 60.3 µm in the sample were useful up to 0.001% on the low content side (Example 14). Particles having a particle diameter of 17.9 and 22 µm were dispersed even at a high content of 5%, and the visibility was also good and could be used as a transparent structure visualizing agent (Examples 13 and 24). In addition, when the PLA0005 concentration was 2%, it was observed that it can be used as a transparent tissue visualizing agent even in particle sizes of any size tested (Examples 5, 12 and 23). In view of the above, it is apparent that the polylactic acid microparticles tested showed an improvement in visibility even at a content of 0.0001 w / v%, and an improvement in visibility even at a content of 0.005 w / v% or more.

5-2 Test Result of Visibility Enhancement Effect of Transparent Tissue Visualizing Agent Containing Soluble Starch:

Referring to Table 5, the soluble starch was ground using an A-O jet mill to obtain particles having a particle diameter of 35.8 mu m. In the particles having a particle size of 35.8 µm, the vitreous could be visualized up to a content of 0.0025 w / v% (Example 26). Therefore, if it is the range of at least 0.0025%-10 w / v%, it can be used as a transparent tissue visualizing agent.

5-3. Test result of visibility improvement effect of transparent tissue visualizing agent containing other high molecular compound:

Referring to Table 6, even in the microparticles composed of PLGA5005, PLA0010 or chitosan, the vitreous was visually recognized at a content of 1 w / v% (Examples 33 to 35). Further, in PLGA5005, the vitreous could be visualized at a content of 0.01% (Example 32).

Test Example 2 Redispersibility Test of Transparent Tissue Visualizing Agent

1.Test Method:

See Table 7. In the same manner as in Test Example 1, the transparent tissue visualizing agents of Examples 36 to 38 were prepared. In a 5 mL syringe, 0.02 g of the powder sample prepared in Example 36, 0.04 g of the powder sample prepared in Example 37, and O.06 g of the powder sample prepared in Example 38 were each mixed with 2 mL of the following dispersion by hand. Shaking was performed and the number of shaking required to disperse the powder in the dispersion was measured.

Dispersion Formulation

0.75 g of sodium chloride

0.16 g of potassium chloride

0.1 g of sodium hydrogen phosphate

Sodium dihydrogen phosphate ... 0.015 g

Hydrochloric acid ·················

Purified water ・ ・ ・ ・ to 100 mL

2.Test result:

In PLA0005 fine particles, by adding mannitol, the number of shakings until dispersing at the time of mixing with the liquid was reduced, and the redispersibility was improved. By further adding povidone to the mixture, the number of shaking required to achieve dispersion was further reduced, further improving redispersibility.

Preparation Example 1 Vial Filling Transparent Visualizer

An aqueous solution containing 1.11 w / v% D-mannitol and 1.11 w / v% povidone K-30 was filtered through a hydrophilic filter having a pore diameter of 0.22 μm to obtain A liquid. 10 w / v% PLA0005 (acetone / ethanol = 4/6) Acetic solution was prepared, filtered through a hydrophobic filter having a pore diameter of 0.22 µm to obtain B solution. In addition, Opeguard MA was made into C liquid.

A liquid and B liquid were mixed as follows at a 9: 1 ratio. That is, while stirring A liquid at 700-800 rpm with the stirrer, B liquid was added at about 100 microliters / second, and PLA0005 was precipitated as microparticles | fine-particles. After the mixture was stirred for about 30 minutes, the aggregated mass was removed by a sieve (diameter of 106 mu m) to obtain a suspension D. It was lyophilized in vials to give powder sample E. The sample was prepared by adding C liquid to the vial containing powder sample E before use.

In addition, lyophilization was operated as follows. That is, suspension D was preserve | saved at -40 degreeC for 6 hours, it frozen, and it pressure-reduced to 100 micrometers Hg or less under -40 degreeC, and dried for 24 hours or more. The temperature was raised by 10 ° C every hour at -40 ° C and continued until + 20 ° C. Further dried at + 20 ° C. and below 100 μm Hg for at least 24 hours.

Preparation Example 2 Double Chamber Type Syringe Filled Transparent Tissue Visualizing Agent

An aqueous solution containing 1.11 w / v% D-mannitol and 1.11 w / v% povidone K-30 was filtered through a hydrophilic filter having a pore diameter of 0.22 μm to obtain F liquid. On the other hand, a 10 w / v% PLA0005 (acetone / ethanol = 4/6) solution was filtered through a hydrophobic filter having a pore size of 0.22 µm to obtain a G liquid. Phosphate buffer (pH 7) containing 0.75% sodium chloride and 0.16% potassium chloride was used as the H liquid.

F liquid and G liquid were mixed as follows at the ratio of 9: 1. That is, G liquid was dripped at about 100 microliters / sec, stirring F liquid at 700-80 rpm in the stirrer, and PLA0005 was precipitated as microparticles | fine-particles. The mixture was stirred for about 40 minutes (10 minutes under reduced pressure), and then the aggregated mass was removed by a sieve (diameter of 106 mu m) to obtain a suspension J. Each 2 mL of this was dispensed to the solid-phase side 3 of the double chamber type syringe 1 which shows the outline in FIG. 1, lyophilized, and powder L was obtained. A rubber stopper provided as a movable separator 2 is sandwiched between the solid phase side 3 and the liquid phase side 4 so that the syringe is filled with 2 mL of liquid H provided as a medium 6 on the liquid side 4. Thereafter, a stopper of piston 8 was inserted to prepare a double chamber-type syringe filled transparent tissue visualization agent. 9 is a peripheral flow path in the front-rear direction in which the inner wall of the double chamber-type syringe 1 is partially recessed. Lyophilization was carried out as follows. That is, suspension J was preserve | saved at -40 degreeC for 6 hours, it frozen, and it pressure-reduced to -100 micrometerHg under -40 degreeC, and dried for 24 hours or more. The temperature was raised by 10 ° C every hour at -40 ° C and continued until + 20 ° C. Furthermore, it dried at +10 degreeC and 100 micrometers Hg or less for 24 hours or more.

The use method of the transparent tissue visualizing agent of this embodiment is as follows. That is, the piston 8 is pushed forward to thereby advance the movable separator 2 by using the pressure generated in the liquid H as the medium sealed on the liquid phase side 4. When the rear side of the movable separator 2 reaches the peripheral flow path 9, the liquid phase side 4 and the solid phase side 3 communicate with each other, and the liquid H is injected into the solid phase side 3. By advancing the piston 8 until it comes in contact with the movable separator 2, all of the liquid H is transferred to the solid-phase side 3 and mixed with the powder L there. After the mixing is completed, the piston 8 is further advanced (with the movable separator 2) to discharge the mixed liquid from the discharge passage 7 to the surgical operation site (via a needle or the like not shown).

Preparation Example 1 Transparent Tissue Visualizing Agent Containing Soluble Starch Fine Particles

By a conventional method, the transparent tissue visualizing agent of the following formulation was prepared. The average particle diameter of the soluble starch microparticles in the formulation was about 50 μm.

Soluble starch ... 1.0 g

Opeguard MA ... to 100 mL total

Preparation Example 2 Chitosan Fine Particle-Containing Transparent Tissue Visualizing Agent

By a conventional method, the transparent tissue visualizing agent of the following formulation was prepared.

1.0 g of chitosan

Opeguard MA ... to 100 mL total

Preparation Example 3 Transparent Tissue Visualizing Agent Containing Polylactic Acid Fine Particles

By the conventional method, the transparent tissue visualizing agent of the following formulation was prepared. The average particle diameter of the polylactic acid microparticles in the formulation was about 0.2-0.3 μm.

1.0 g of polylactic acid

Opeguard MA ... to 100 mL total

Preparation Example 4 Lactate-Glycolic Acid Copolymer Fine Particle-Containing Transparent Tissue Visualizing Agent

By the conventional method, the transparent tissue visualizing agent of the following formulation was prepared. The average particle diameter of the lactic acid-glycolic acid copolymer microparticles in the formulation was about 0.06 to 0.07 μm.

Lactic acid-glycolic acid copolymer ... 1.0 g

Opeguard MA ・ ・ ・ ・ 100 ml total

Preparation Example 5 Transparent Tissue Visualizing Agent Containing Soluble Starch Fine Particles

By a conventional method, the transparent tissue visualizing agent of the following formulation was prepared.

Soluble starch: 1.0 g

Sodium dihydrogen phosphate dihydrate ... 0.10 g

Sodium chloride 0.9 g

Sodium Hydroxide ·········

Purified water ・ ・ ・ ・ to 100 mL

pH ··············· 7.0

Preparation Example 6 Transparent Tissue Visualizing Agent Containing Chitosan Fine Particles

By the conventional method, the transparent tissue visualizing agent of the following formulation was prepared.

0.10 g of chitosan

Sodium chloride0.55 g

0.16 g of potassium chloride

Dry sodium carbonate ... 0.06 g

Sodium hydrogen phosphate ... 0.18 g

Boric acid ... 1.2 g

Borax ········

Sterilized purified water ・ ・ ・ ・ to 100 mL

pH 7.3

Preparation Example 7 Transparent Tissue Visualizing Agent Containing Polylactic Acid Fine Particles

By the conventional method, the transparent tissue visualizing agent of the following formulation was prepared.

100 g of polylactic acid

Sodium dihydrogen phosphate dihydrate ... 0.1 g

Sodium chloride 0.9 g

Sodium Hydroxide ·········

0.1 g sodium edetate

Sterilized purified water ・ ・ ・ ・ to 100 mL

pH ··············· 7.0

Preparation Example 8 Transparent Tissue Visualizing Agent Containing Lactic Acid-Glycolic Acid Copolymer Fine Particles

By the conventional method, the transparent tissue visualizing agent of the following formulation was prepared.

Lactic acid-glycolic acid copolymer0.10 g

Sodium chloride0.55 g

0.16 g of potassium chloride

Dry Sodium Carbonate ·········· 0.06 g

Sodium hydrogen phosphate ··········· 0.18 g

Boric acid ... 1.2 g

Borax ············

Hydroxypropylmethylcellulose ... 0.1 g

Sterilized purified water ・ ・ ・ ・ to 100 mL

pH ························· 7

The present invention as defined above significantly improves the visibility of difficult-to-visible transparent tissues in the surgical area, without causing unnecessary reactions or side effects to the living body, and in the surgical operations of the vitreous, lens, and corneas, which are transparent tissues of the eyes, The surgery is made easier, and the certainty of the surgical operation purpose is made easier.

Claims (20)

A transparent tissue visualization agent designed to enhance visibility by contacting transparent tissues of the eye, including microparticles of a polymeric compound. The transparent tissue visualizing agent according to claim 1, wherein 1 g of the fine particles cannot be completely dissolved in less than 30 mL of water at 20 ° C within 30 minutes. The transparent tissue visualizing agent according to claim 1 or 2, wherein the fine particles have an average particle diameter of 0.01 to 500 µm. The transparent tissue visualizing agent according to any one of claims 1 to 3, wherein the polymer compound is a biodegradable polymer compound and / or a water-soluble polymer compound. The transparent tissue visualization agent according to claim 4, wherein the biodegradable polymer compound is selected from fatty acid polyesters, polysaccharides and derivatives thereof. The transparent tissue visualizing agent according to claim 4, wherein the water-soluble high molecular compound is selected from an acrylic polymer compound and a styrene polymer compound. The method according to any one of claims 1 to 3, wherein the polymer compound is starch, soluble starch, pregelatinized starch, cellulose, acetylcellulose, hydroxypropyl methylcellulose, chitosan, chitin, dextran, polylactic acid, poly Glycolic acid, lactic acid-glycolic acid copolymer, polyhydroxybutyric acid, polyhydroxy valeric acid, polycaprolactone, hydroxybutyric acid-glycolic acid copolymer, lactic acid-caprolactone copolymer, polyethylene succinate, and polybutylene succinate Transparent tissue visualizing agent selected from the group consisting of nate, sodium polyacrylate, sodium methacrylate and sodium polystyrenesulfonate. The transparent tissue visualizing agent according to claim 7, wherein the polymer compound has an average molecular weight of 500 to 200000. 10. The transparent tissue visualizing agent of claim 8 further comprising a polyvinyl compound and / or a polyol. The transparent tissue visualization agent according to claim 9, which contains 0.05 to 10 parts by weight of the polyvinyl compound and / or 0.05 to 10 parts by weight of the polyol based on 1 part by weight of the fine particles. The transparent tissue visualizing agent according to claim 9 or 10, wherein the polyvinyl compound is polyvinyl pyrrolidone and / or polyvinyl alcohol. The transparent tissue visualizing agent according to claim 9 or 10, wherein the polyol is mannitol. The transparent tissue visualizing agent according to any one of claims 1 to 12, further comprising an aqueous medium, wherein the fine particles are dispersed in the aqueous medium. The transparent tissue visualization agent according to claim 13, wherein the content of the fine particles in the transparent tissue visualization agent is 0.005 to 10 w / v%. The transparent tissue visualizing agent according to claim 13 or 14, further comprising a polyvinyl-based compound and / or a polyol. The transparent tissue visualizing agent according to claim 15, which contains a polyvinyl-based compound at a concentration of 0.1 to 5 w / v% and / or a polyol at a concentration of 0.1 to 5 w / v%. 17. The clear tissue visualizing agent according to any one of claims 1 to 16, which is an injectable preparation. A transparent tissue visualization agent comprising a single mixing and discharging means enclosed in a state in which the powder composition comprising the fine particles according to any one of claims 1 to 12 and the aqueous medium are separated from each other so as to be in contact with each other. A method for improving the visibility of transparent tissue of an eye comprising contacting the composition comprising the microparticles of any one of claims 1 to 17 with the transparent tissue of the eye. Use of the microparticle of any one of claims 2 to 8 for the preparation of a clear tissue visualizing agent designed to contact and visualize the clear tissue of the eye.

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