Eye drop composition comprising novel molecular assembly of axitinib and preparation method therefor

Abstract

The present invention relates to a composition comprising a molecular assembly in which axitinib is physically associated, and to a preparation method therefor.

Description

Ophthalmic composition comprising novel molecular complex of axitinib and method for preparing the same

[0001] The present invention relates to a composition comprising a novel molecular aggregate of axitinib, and more specifically, to an eye drop composition having an excellent effect in treating and preventing wet macular degeneration by manufacturing a composition comprising a novel molecular aggregate by a bottom-up method using shear stress.

[0002] Axitinib is known as an oral, selective inhibitor of vascular endothelial growth factor (VEGF) receptors 1, 2, and 3, used to treat metastatic renal cell carcinoma (mRCC). It is also used as a drug that blocks VEGF signaling in the treatment of wet macular degeneration. Regeneron Pharmaceuticals' EYELEA is an intraocular injection for the treatment of wet macular degeneration. ® It is known that products injected into the eye have side effects such as bleeding at the injection site, eye pain, cataracts, vitreous detachment, vitreous floaters, and increased intraocular pressure.

[0003] Recently, ongoing research has focused on the potential of cyclodextrin-based ophthalmic compositions as a novel treatment for wet age-related macular degeneration, promising potential for delivering drugs to the posterior chamber of the eye to lower retinal cholesterol. Cyclodextrin is believed to offer an economical, safe, and effective route for reducing the effects of retinal aging. However, excessively high cyclodextrin concentrations have been associated with swelling of the upper and lower eyelids, some ocular congestion, and nictitating membrane edema in animal models. Furthermore, low cyclodextrin concentrations relative to the active ingredient can compromise the stability of the composition.

[0004] Accordingly, there was a need for an eye drop composition having an excellent treatment and prevention effect for wet macular degeneration by manufacturing a novel molecular complex of axitinib and an eye drop composition containing the same, which is easy to manufacture and has stability.

[0005] (Patent Document 1) US 9,968,603 B2

[0006] The present invention aims to provide an eye drop composition that is effective in treating wet macular degeneration in vivo. More specifically, the present invention aims to provide an eye drop composition that is easy to manufacture and has stability, thereby exhibiting excellent therapeutic effects for wet macular degeneration.

[0007] To achieve the above purpose,

[0008] The present invention provides a composition characterized by comprising a molecular aggregate physically bound to axitinib; a solubilizer; a stabilizer; and an additive.

[0009] In addition, the present invention provides a method for preparing a composition, comprising the steps of: (a) adding an acid to a molecular complex to which axitinib is physically bound to dissolve; (b) adding a solubilizer to the solution and stirring the solution to prepare a first solution; (c) solubilizing a stabilizer in water to prepare a second solution; (d) mixing the first solution and the second solution; (e) adding an additive including a buffer and an osmotic pressure regulator as active ingredients to the solution prepared in step (d), adjusting the pH to 6 to 8 with a pH regulator, and then adding water to the solution to prepare the pH; and (f) sterilizing the solution prepared in step (e).

[0010] The ophthalmic composition comprising the novel molecular complex of axitinib according to the present invention is a composition having an easy manufacturing method and stability, and has an excellent treatment effect on wet macular degeneration by delivering the active substance well to the posterior part of the eye in the form of ophthalmic drops, and does not cause problems such as swelling of the upper / lower eyelids, bloodshot eyes in some parts of the eye, and edema of the nictitating membrane, which occur when combinations of existing known compositions fail to show effects in vivo.

[0011] Figure 1 is a graph showing the results of an in-vivo drug efficacy evaluation of an exemplary composition of the present invention, showing a decrease in the area of ​​neovascularization and the number of spots in a laser-induced monkey model.

[0012] Figure 2 is a graph showing the results of an in-vivo drug efficacy evaluation of an exemplary composition of the present invention, showing a reduction in retinal lesion thickness in a laser-induced monkey model.

[0013] The present invention provides a composition comprising a molecular aggregate physically bound to axitinib. Furthermore, the composition can ultimately be used as an ophthalmic composition, and can be used for the prevention or treatment of wet macular degeneration.

[0014] This is explained in more detail below.

[0015]

[0016] The composition according to the present invention comprises a molecular aggregate to which axitinib is physically bound; a solubilizer; and a stabilizer.

[0017] The composition according to the present invention comprises a molecular aggregate to which axitinib is physically bound.

[0018] The above molecular complex to which axitinib is physically bound may be a molecular complex to which axitinib is a compound of the following chemical formula 1.

[0019] [Chemical Formula 1]

[0020]

[0021] The above molecular complex physically bound to the axitinib has an X-ray powder diffraction spectrum of the molecular complex having X-ray diffraction peaks at diffraction angles 2θ of 24.99°±0.1° and 26.32°±0.1°.

[0022] In addition, the molecular assembly to which the axitinib is physically bound can have a DSC profile having a glass transition at a single endothermic temperature of 220.4±2.0°C when measured under differential scanning calorimetry (DSC) conditions of a heating rate of 10°C / min, 99.999% N2, and 30-250°C. That is, while conventional general axitinib exhibits glass transitions at two endothermic temperatures of about 212.5°C and about 220.6°C based on the DSC profile, the molecular assembly to which the axitinib is physically bound of the present invention has a difference in that it has a DSC profile characterized by a glass transition at a single endothermic temperature of 220.4±2.0°C.

[0023] In addition, the crystals of the molecular aggregate to which the above-mentioned axitinib is physically bound may have an average particle size of 2.0 to 15 μm, preferably 3.0 μm or more, 5.0 μm or more, and 13.0 μm or less, 10.0 μm or less. If the average particle size of the molecular aggregate crystals exceeds 15.0 μm, there is a problem that dispersibility is reduced and transparency and transmittance are reduced. In addition, if the average particle size of the molecular aggregate crystals is less than 2.0 μm, there is a problem that manufacturing is difficult and performance is not expressed.

[0024]

[0025] In addition, the molecular complex to which the axitinib is physically bound may have an aspect ratio value of 0.3 to 1.0. That is, unlike conventional general axitinibs that have an aspect ratio value of less than 0.3 and have an elongated rod-shaped shape, the molecular complex to which the axitinib is physically bound of the present invention has a physically bound structure of pure axitinib, and thus exhibits the characteristic of having an aspect ratio value of 0.3 or more and has a difference in that it has a relatively round shape.

[0026] The molecular complex to which the above-mentioned axitinib is physically bound may have an aspect ratio of 0.3 or more, 0.4 or more, 0.5 or more, 0.6 or more, 0.7 or more, 0.8 or more, 0.9 or more, and may have an aspect ratio of 1.0 or less, 0.9 or less, 0.8 or less.

[0027] In the present invention, the aspect ratio of the particle can be determined by measuring the length and thickness of the particle using any suitable measuring technique, preferably a dynamic image analysis method performed according to the ISO 13322-2:2006 standard, and calculating the aspect ratio from the measured dimensions of the particle as described above.

[0028]

[0029] In addition, the molecular complex in which the above-mentioned axitinib is physically bound may have a solubility of 3.0 mg / mL or more in a solubility concentration at pH 1 and 0.1 mg / mL or more in a solubility concentration at pH 2.

[0030] Specifically, the molecular complex to which the above-mentioned axitinib is physically bound may have a solubility concentration at pH 1 of 3.0 mg / mL or more, 3.5 mg / mL or more, 4.0 mg / mL or more, or 4.3 mg / mL or more, and the upper limit may be 10.0 mg / mL or less, although there is no particular limitation thereon.

[0031] In addition, specifically, the molecular complex to which the above-mentioned axitinib is physically bound may have a solubility concentration at pH 2 of 0.1 mg / mL or more, 0.3 mg / mL or more, 0.5 mg / mL or more, 1.0 mg / mL or more, 1.5 mg / mL or more, or 1.7 mg / mL or more, and the upper limit may be 5.0 mg / mL or less, although there is no particular limitation thereon.

[0032]

[0033] The molecular complex in which the above axitinib is physically bound may have a solubility 1.5 or 2 times higher than that of the original axitinib itself.

[0034] The above molecular complex in which the axitinib is physically bound can be prepared by applying shear stress to a solution containing axitinib or a salt of axitinib, which is a precursor of the above structure.

[0035] The shear stress applied to the solution containing axitinib, which is a precursor of the above structure, may be either mechanical shear stress or ultrasonic application.

[0036] The above mechanical shear stress may be applied by passing the solution through a silica-filled column or filter paper. The mechanical shear stress is described in detail below.

[0037] According to one embodiment of the present invention, the mechanical shear stress may be applied by passing a solution containing axitinib through a column filled with silica. When the solution containing axitinib passes through a column filled with silica or the like, the axitinib is subjected to very high shear stress by passing through a physically narrow region.

[0038] The above silica may be spherical or angular, but is not limited to its shape.

[0039] The size of the silica may be 0.01 to 100 μm, preferably 0.1 to 10 μm, and more preferably 2.5 to 3.7 μm. When the size of the silica is less than 0.01 μm or greater than 100 μm, even if the solution containing the axitinib passes through a column filled with silica, shear stress is not applied, so there may be no change in the structure.

[0040] A negative pressure of 0.1 bar to 1.0 bar or 0.2 bar to 0.9 bar can be applied to the lower portion of the silica-filled column. When the negative pressure applied to the lower portion of the silica-filled column is less than 0.1 bar, the time required for the solution containing axitinib to pass through the column increases, thereby delaying the manufacturing time of axitinib according to the present invention. In addition, when the negative pressure applied to the lower portion of the silica-filled column is more than 1.0 bar, the time required for the solution containing axitinib to pass through the column decreases, thereby shortening the manufacturing time of axitinib according to the present invention. However, since additional pump equipment is required, the manufacturing cost may increase.

[0041] According to another embodiment of the present invention, the mechanical shear stress may be applied by passing the solution containing the axitinib through one or more filter papers. Passing through the one or more filter papers physically exposes the axitinib to very high shear stress by passing through a narrow area.

[0042] The filter paper may be a single filter paper or two or more filter papers. If the filter paper is two or more filter papers, the filter papers may be arranged in a stacked manner. If the filter paper is two or more filter papers, a higher shear stress may be provided than that of a single filter paper.

[0043] The pore size of the filter paper may be 0.1 to 5.0 microns or 0.3 to 4.5 microns. When the pore size of the filter paper is less than 0.1 micron, the amount of the solution containing axitinib passing through or filtering the filter paper may be too small, so that the production speed of axitinib according to the present invention may be reduced, and when the pore size of the filter paper is greater than 5.0 microns, the solution containing axitinib may simply pass through the filter paper, so that shear stress may not be effectively applied.

[0044]

[0045] The composition according to the present invention comprises a solubilizer.

[0046] In the present invention, a solubilizer is used to increase the solubility of a drug, and the solubilizer used in the present invention is alphacyclodextrin (alpha-CD), beta-cyclodextrin (beta-CD), gamma-cyclodextrin (gamma-CD), 2-hydroxypropyl-beta-cyclodextrin (HP-beta-CD), sulfobutylether beta-cyclodextrin (SBE-beta-CD); 2-hydroxypropyl-alpha-cyclodextrin; randomly methylated beta-cyclodextrin; 2-O-methyl- beta-cyclodextrin; 2,6-di-O-methyl-beta-cyclodextrin; heptakis(2,3,6-tri-O-methyl)-beta- cyclodextrin; carboxymethyl-beta-cyclodextrin; carboxyethyl-beta-cyclodextrin; hydroxyethyl- beta-cyclodextrin; maltosyl-beta-cyclodextrin; 3,6-(N,N,N-trimethylammonium)propyl-beta-cyclodextrin; acetyl-beta-cyclodextrin; 2,6-di-O-methyl-gamma-cyclodextrin; 2-hydroxypropyl-gamma-cyclodextrin; or sulfobutylether gamma-cyclodextrin;optionally, wherein the cyclodextrin may include at least one selected from the group consisting of alpha-cyclodextrin (alpha-CD), beta-cyclodextrin (beta-CD), gamma-cyclodextrin (gamma-CD), 2-hydroxypropyl-beta-cyclodextrin (HP-beta-CD), or sulfobutylated-beta-cyclodextrin (SBE-beta-CD) and cyclodextrin derivatives;

[0047] The above solubilizing agent is a cyclodextrin derivative. Conventional cyclodextrins, when used as a component of a composition, have been used in a form that encapsulates (complexes) with drugs. However, in the present invention, cyclodextrin and cyclodextrin derivatives can be used as a component of the composition for simple solubilization of drugs, rather than encapsulation.

[0048]

[0049] The composition according to the present invention comprises a stabilizer.

[0050] In the present invention, a stabilizer is used to maintain the stability of a drug and preserve its efficacy, and may exist in various forms. It may be used to increase the solubility of a selected drug depending on the characteristics and use of the drug. The stabilizer of the present invention may include at least one selected from the group consisting of methylcellulose (MC), hydroxymethyl cellulose (HMC), carboxymethylcellulose (CMC), hydroxypropyl methylcellulose (HPMC), hydroxyethyl cellulose (HEC), and cellulose derivatives.

[0051] The composition according to the present invention may further include an additive, and any additive that is normally included may be used without particular limitation, and preferably, an additive including at least one of a buffer; an osmotic pressure regulator; and a pH regulator may be used.

[0052] The above buffer plays a role in maintaining the stability and efficacy of the drug by maintaining a biocompatible pH (hydrogen ion concentration) range at a certain level, and the composition according to the present invention may specifically include at least one selected from the group consisting of citric acid, phosphoric acid, boric acid, tartaric acid, acetic acid, and amino acids as the above buffer.

[0053] The buffer is an osmotic pressure regulator that maintains the appropriate melting point (dissolution point) of the drug and controls the size and dispersion of drug particles to help better absorption upon administration. The composition according to the present invention may specifically include at least one selected from the group consisting of sorbitol, mannitol, dextrose, sucrose, and glycerin as the osmotic pressure regulator.

[0054] The above pH regulator serves to provide an appropriate chemical environment by adjusting the pH value of the drug, and the composition according to the present invention can specifically use an alkalizing agent as the pH regulator.

[0055]

[0056] The composition according to the present invention comprises a molecular aggregate physically bound to axitinib; a solubilizer; a stabilizer;

[0057] The above molecular aggregate may comprise, based on the entire composition, 0.02 to 0.12 w / v% of the molecular aggregate to which axitinib is physically bound; 2 to 12 w / v% of the solubilizer; and 0.01 to 1.0 w / v% of the stabilizer. If the content of the molecular aggregate to which axitinib is physically bound is lower than 0.02 w / v%, the effect of using the drug may be minimal, and if the content is higher than 0.12 w / v%, a problem such as swelling of the upper / lower eyelids may occur.

[0058] The composition according to the present invention may further include additives, and the additives may include a buffer and an osmotic pressure regulator, wherein, based on the entire composition, the buffer may be included in an amount of 0.5 to 1.5 w / v%; and the osmotic pressure regulator may be included in an amount of 0.1 to 5 w / v%.

[0059]

[0060] The composition according to the present invention may include a pH regulator as an additive, and the pH can be adjusted to 6 to 8 using the pH regulator.

[0061]

[0062] The composition according to the present invention may be for eye drops.

[0063] The composition according to the present invention may have an appropriate viscosity for use as an eye drop, and a specific viscosity may be 100 m·Pas or less, a viscosity may be 80 m·Pas or less, a viscosity may be 60 m·Pas or less, or a viscosity may be 40 m·Pas or less. The lower limit of the viscosity is not particularly limited, but may typically be 5 m·Pas or more. If the viscosity of the eye drop composition exceeds 100 m·Pas, it may be very sticky, which may reduce the comfort of the eye drop.

[0064]

[0065] The composition according to the present invention may be a pharmaceutical composition for preventing or treating wet macular degeneration.

[0066] The macula, located at the center of the retina, plays a crucial role in vision. Wet macular degeneration (AMD) is a disease in which the macula, the central area of ​​the retina, is damaged due to aging. It is primarily found in patients with eyelid symptoms. Wet AMD primarily affects the elderly and is one of the most common causes of vision loss.

[0067] The above macular degeneration is broadly classified into dry macular degeneration and wet macular degeneration. Wet macular degeneration is characterized by the formation of abnormal blood vessels in the retina or their abnormal exposure. These changes can include vascular leakage, neovascularization, and the formation of new blood vessels on the retina, and these processes can lead to vision loss.

[0068] The composition according to the present invention can prevent or treat the symptoms of such wet macular degeneration.

[0069]

[0070] The present invention provides a method for preparing a composition, comprising: (a) adding an acid to a molecular complex in which axitinib is physically bound to dissolve the complex; (b) adding a solubilizer to the solution and stirring the solution to prepare a first solution; (c) solubilizing a stabilizer in water to prepare a second solution; (d) mixing the first solution and the second solution; (e) adding an additive including a buffer and an osmotic pressure regulator as active ingredients to the solution prepared in step (d), adjusting the pH to 6 to 8 with a pH regulator, and then adding water to the solution to be labeled; and (f) sterilizing the solution prepared in step (e).

[0071]

[0072] In the above manufacturing method, the contents of the molecular aggregate to which axitinib is physically bound, the solubilizer, the stabilizer, the buffer, and the osmotic pressure regulator are the same as those described above in the composition.

[0073]

[0074] The characteristics of the above manufacturing method can be specified as follows, but are not limited thereto.

[0075] In step (a), an acidifying agent such as hydrochloric acid can be added to water to obtain a solution in which the molecular aggregates physically bound to axitinib are dissolved or suspended in the acidic aqueous solution. In addition, when the acidifying agent is mixed with water, the temperature of the aqueous solution increases, so that it can be cooled to 20 to 30°C before use.

[0076] (c) When using a cellulose derivative as a stabilizer in step (c), a device commonly used to dissolve cellulose in water, such as a stirrer or homogenizer, can be used to heat the cellulose derivative to a temperature of 60°C or higher to solubilize the cellulose derivative or to dissolve it by stirring.

[0077] The order of steps (b) and (c) above can be changed and used, and it is possible to add the aqueous solution of step (b) to the aqueous solution of (c) and then proceed to the next step.

[0078] (d) When proceeding with step, it can be done at a temperature below 30℃ and with shading.

[0079] Sterilization in step (f) can be performed by aseptic filtration, and sterilization can be performed by filtration with a filter.

[0080]

[0081] Hereinafter, the present invention will be described in more detail through examples. It will be understood that the present invention is not limited to these examples.

[0082]

[0083] [Manufacturing example]

[0084] 1. Preparation of molecular assembly (SCAI-005)

[0085] A solution of axitinib with a concentration of approximately 0.1% was prepared by dissolving 16.0 g of axitinib [Shilpa, India] in 16.0 kg of ethanol (94.5% ethanol, Samjeon).

[0086] 270 g of SYLOID 244FP (GRACE, USA) was wetted with 4.32 kg of ethanol (94.5%, Ethanol, Samsung), and a 1.0 μm paper filter was attached to a 350 mm diameter Nutsche filter to prepare a SYLOID 244FP column with a height of approximately 1.4 cm by pouring the SYLOID 244FP solution wetted with ethanol into the Nutsche filter.

[0087] The SYLOID 244FP column was made solid by adding 1.08 kg of 94.5% ethanol prepared on the column using a vacuum. The prepared axitinib solution was added, and an additional 3.24 kg of 94.5% ethanol was passed through the column to recover the axitinib remaining in the SYLOID 244FP. The weight of the axitinib effluent at this time was approximately 21.86 kg.

[0088] The axitinib effluent was filtered using a 0.45 μm PVDF membrane filter and concentrated to a concentration of 3.0 mg / g using a rotary vacuum evaporator. After concentration was completed, the axitinib concentrate was filtered using a 0.2 μm PVDF membrane filter.

[0089] 53.0 kg of purified water was added to a 100 L reactor, and the prepared axitinib concentrate was slowly added while rapidly stirring the purified water. After the addition was complete, stirring was continued for an additional 30 minutes. The mixture was filtered using a 1.0 μm paper filter.

[0090] The filtered cake was decompressed under vacuum for 30 minutes and dried using nitrogen for 2 hours. In addition, it was dried in a vacuum oven at 25°C for 38 hours to obtain 14.07 g (Y. 88%) of axitinib as a white powder.

[0091]

[0092] Manufacturing Experimental Example 1. X-ray Diffraction Analysis (X-ray Diffractometer, XRD) of the New Axitinib Polymorph (SCAI-Form)

[0093] When the reagent is placed in the specimen holder, pressed with a glass rod to form a charge in the charging section and tested according to the powder X-ray diffraction measurement method in the general test methods of the Korean Pharmacopoeia, it exhibits a crystalline form.

[0094] - Operating conditions

[0095] Condition Actual measurement condition Condition Actual measurement condition

[0096]

[0097] Powder X-ray diffraction patterns of various polymorphic forms were performed on a Rigaku Miniflex600 using copper radiation (CuKα, wavelength: 1.5406 Å). The tube voltage and current were set to 40 kV and 15 mA, respectively. The divergence and scatter slits were set to 8.0 mm, and the receiving slit was set to 13.0 mm. The diffracted radiation was detected with a D / teX Ultra2. A theta-2 theta continuous scan at 2.0 degrees / min (1 s / 0.03 degree step) was used from 3.0 to 60 degrees 2θ. An alumina standard was analyzed to verify instrument alignment. The data were collected and analyzed using SmartLab Studio Ⅱ.

[0098]

[0099] Powder X-ray diffraction patterns were measured on a Rigaku Miniflex 600 using copper radiation (CuKα, wavelength: 1.54056 Å). The tube voltage and current were set to 40 kV and 15 mA, respectively. The divergence and scattering slits were set to 8.0 mm, and the receiving slit was set to 13.0 mm. The diffracted radiation was detected with a D / teX Ultra2. A theta-2 theta continuous scan at 2.0 degrees / min (1 s / 0.03 degrees) was used over a 2θ range of 3.0 to 60 degrees. An alumina standard was analyzed to verify instrument alignment. Data collection and analysis were performed using SmartLab Studio Ⅱ.

[0100]

[0101] - 2 theta and relative intensity of XRD results of API and SCAI-Form

[0102] APISCAI-Form(AXTEN03A)Anglerelativeintensity(%)Anglerelativeintensity(%)8.19102.708. 8453.211.924.2311.99133.114.743.9314.602615.3420.1315.2451.915.5643.4115.7076.617.45 57.1917.7127.619.394.7219.3358.120.698.4120.6558.821.409.2121.7085.123.2944.4623.2353.424.0155.3924.193825.9610024.99(25.0)10026.243.7526.32(26.3)38.827.788.1327.5911.9

[0103]

[0104] Manufacturing Experimental Example 2. Differential Scanning Calorimetry (DSC) of the Novel Axitinib Polymorph (SCAI-Form)

[0105] Measurements were made using the temperature increase program [Table 3] in the differential scanning calorimeter device. At this time, the sample amount is recommended to be 4.0 mg or less, the environment inside the device is maintained as nitrogen, and the nitrogen flow rate is 10 mL / min.

[0106] RateTargetHoldRecord20 ℃ / min100 ℃20 min10 ℃ / min30 ℃5 min-10 ℃ / min250 ℃5 min√

[0107] Peak temperature and ΔH of API and SCAI-Form

[0108] APISCAI-Form(AXTEN03A)peak temp.(℃)212.5220.4220.6ΔH(J / g)129.1131.7

[0109]

[0110] Manufacturing Experimental Example 3. Scanning Electron Microscope (SEM) of the Novel Axitinib Polymorph (SCAI-Form)

[0111] Measurement conditions

[0112] Powder samples were placed on carbon tape fixed to an aluminum stub. The samples were scanned in a FE-SEM using a JSM-IT800, Jeol. Images were acquired at an accelerating voltage of 1.00 kV using a secondary electron detector.

[0113]

[0114] The crystal structures of Axitinib API and SCAI-Form were analyzed using the above measurement method. As shown in Table 5 below, the crystals of the molecular aggregate were confirmed to have an average particle size of 2–15 μm.

[0115] - Particle diameter

[0116] Distinctive diameter (length μm) average value 6.68 minimum value 3.29 maximum value 12.84

[0117]

[0118] Manufacturing Experimental Example 4. Solubility of the Novel Axitinib Polymorph (SCAI-Form)

[0119] pH 1 Solubility Measurement Method: Stir for 15 minutes at a concentration of 5 mg / mL, filter, dilute 10 times with DW, and conduct analysis. (The value calculated as [measured value x 10] is reported in the results.)

[0120] pH 2 Solubility Measurement Method: After stirring for 15 minutes at a concentration of 1 mg / mL, filtering was performed for analysis.

[0121] - Solubility

[0122] Distinction Batch No. pH 1 Dissolution Concentration (mg / mL) pH 2 Dissolution Concentration (mg / mL) API Axitinib API (Shilpa) 2.6610.073 SCAI-Form AXTEN03A4.3630.171

[0123] As above, it can be seen that the solubility of SCAI-Form is about twice as high as that of API.

[0124]

[0125] [Example]

[0126] 1. Manufacturing method and composition of eye drops containing SCAI-005

[0127] Eye drops of Examples 1 to 15 were prepared by the following method under the conditions of Tables 7 and 8 below.

[0128] 1) After dissolving SCAI-005 by adding hydrochloric acid to water, hydroxypropyl beta-cyclodextrin (HP-beta-CD) or hydroxypropyl gamma-cyclodextrin (HP-gamma-CD) was added as a solubilizer and stirred.

[0129] 2) The stabilizer was dissolved in water.

[0130] 3) Mixed with the above solutions (1) and (2).

[0131] 4) Additives (buffer, osmotic pressure regulator) as active ingredients to the above solution (3), adjust the pH to about 7 with an alkalizing agent, and then add water to the solution to mark the volume.

[0132] Example 1 Example 2 Example 3 Example 4 Example 5 Example 6 Example 7 Example 8 Batch No. (JE30)-03 (JE30)-27 (JE30)-29 (JE30)-30 (JE30)-30-1 (JE82-6)-66 (JE82-6)-72 (JE91A)-100 SCAI-005 0.02% HP-β-Cyclodextrin 1% 1% 1% 1% 2% 2.4% 2.3% HP-γ-Cyclodextrin--1% HPMC 2910 (4000 mPas)-0.1% 0.1% 0.10.10.10.1 Povidone K30 1% 1% 2.5%--Povidone K90-----Citric buffer 0.728% 1N HCl 1N NaOH

[0133] Example 9 Example 10 Example 11 Example 12 Example 13 Example 14 Example 15 Batch No. (JE91A)-101 (JE91A)-110 (JE91A)-102 (JE91A)-103 (JE91A)-112 (JE88)-94 (JE91A)-104 SCAI-005 0.02% 0.04% 0.08% HP-β-Cyclodextrin 2.8% 4% 5.2% 5.6% 7% 8% 9.6% - HPMC 2910 (4000 mPas) 0.1% Citric buffer 0.728% 1N HCl q.v. 1N NaOH q.v.

[0134]

[0135] [Experimental Example]

[0136] The following experiments were conducted on the eye drop compositions manufactured in Examples 1 to 15 above.

[0137] Experimental Example 1. Stability Experiment (Accelerated Conditions)

[0138] When an accelerated test is conducted according to the Korean Pharmacopoeia, the product must be stable for up to 6 months depending on the accelerated test conditions. After manufacturing Examples 1 to 15, the appearance was visually confirmed according to the accelerated test, and changes in content were measured using liquid chromatography. The results are shown in Table 9.

[0139] Initial accelerated test 1 week (40±2°C, 25% or less RH) Accelerated test 3 months (40±2°C, 25% or less RH) Accelerated test 6 months (40±2°C, 25% or less RH) Content properties Content properties Content properties Content properties Example 199.09% Colorless, transparent 27.80% Precipitation----Example 2102.31% Colorless, transparent 75.72% Precipitation----Example 3102.35% Colorless, transparent 73.79% Precipitation----Example 4101.04% Colorless, transparent 81.30% Precipitation----Example 5-Precipitation------Example 6101.64% Colorless, transparent 100.90% Colorless, transparent Precipitation---Example 7100.56% Colorless transparent 100.52% Colorless transparent 75.68% Precipitation--Example 8100.45% Colorless transparent-colorless transparent 101.90% Colorless transparent-precipitation Example 9101.26% Colorless transparent-colorless transparent 102.98% Colorless transparent 97.27% Colorless transparent Example 1098.60% Colorless transparent-colorless transparent-precipitation--Example 1199.90% Colorless transparent-colorless transparent 102.03% Colorless transparent 98.29% Precipitation Example 1299.61% Colorless transparent-colorless transparent 101.58% Colorless transparent 102.85% Colorless transparent Example 13100.99% Colorless transparent - Colorless transparent - Precipitation - Example 1499.38% Colorless transparent - Colorless transparent 98.85% Colorless transparent 103.27% Colorless transparent Example 1599.38% Colorless transparent - Colorless transparent 101.41% Colorless transparent 106.05% Colorless transparent

[0140] As can be seen from Table 9, for Examples 1 to 8, 10, 11 and 13, it was confirmed that the stability was poor under accelerated test conditions, and in the results of Examples 9, 12, 14 and 15, it was confirmed that the amounts of SCAI-005 and hydroxybetacyclodextrin were stabilized from a certain concentration.

[0141]

[0142] Experimental Example 2. In-vivoPharmakokinetics of the SCAI-005

[0143] This experiment used a laser to induce neovascularization by damaging the choroid of both eyes in a monkey animal model, then administered a drug and performed fundus fluorescein angiography (FFA) and optical coherence tomography (OCT) to measure the area of ​​neovascularization in the damaged area, the number of spots, and the thickness of retinal lesions, thereby comparing and evaluating the efficacy.

[0144]

[0145] Laser photocoagulation and group assignment

[0146] Experimental CNV in monkey eyes was induced using a laser photocoagulation system (Vitra 532 nm, Quantel Medical, France).

[0147] Laser parameters: Laser wavelength 532 nm; spot size 50 μm; energy 600 - 800 mW; exposure time 0.1 s.

[0148] A total of eight rhesus monkeys were used to induce CNV. After anesthetizing the animals with an intramuscular injection of ketamine hydrochloride (20 mg / kg) and dexmedetomidine hydrochloride (0.03 mg / kg), each eye was subjected to 8–9 laser burns at approximately one disc diameter from the macula. Care was taken to avoid targeting the macula. The laser power used was determined by the morphology of the subretinal blebs, indicating perforation of the Bruch's membrane. If no blebs formed, higher energy was applied to the next point. Fundus color photography (Retinal Camera TRC-50DX, Topcon Healthcare, Japan) was performed immediately after laser photocoagulation to observe the laser spots. Fundus fluorescein angiography (FFA) was performed 12 days after laser induction. Modeling was considered successful if at least one point of fluorescein leakage of grade III and / or IV was observed. Animals were enrolled in treatment and randomly divided into two groups based on the total area of ​​leakage of grade III and IV laser spots (Tables 10 and 11). Monkeys not included were returned to the Prime Animal Care Unit.

[0149] GroupTest ArticleAmount of monkey (eyes)DosageDose volumeFrequencyRoute of DosingVehicleVehicle3(6)1 drop50 μL / drop4 times / dayfor 28 daysTopicaladministrationTest article Example 15:SCAI-005 (0.08%) Ophthalmic solution3(6)1 drop50 μL / drop4 times / dayfor 28 daysTopicaladministration

[0150] -Grouping

[0151] ArticleLot NumberStorage conditionPhysical propertiesConcentrationExample 15:SCAI-005 (0.08%) Ophthalmic solutionAXTSW(JE91A)-1072~8℃Clear, Colorless solutionAxitinib 0.08%(0.8mg / ml)VehicleAXTSW(00)-0182~8℃Clear, Colorless solution-

[0152] Evaluation Procedure: Anesthesia and pupil dilation

[0153] Before image acquisition, animals were anesthetized with an intramuscular injection of ketamine hydrochloride (20 mg / kg) and dexmedetomidine hydrochloride (0.03 mg / kg). To dilate the pupils, two drops of tropicamide phenylephrine ophthalmic solution were instilled into each eye. After anesthesia, the animals were kept in a darkened room until the pupil diameter exceeded 6 mm. A self-sustaining eyelid spectrophotometer was placed on the eyes.

[0154]

[0155] Fundus photography (FP)

[0156] Retinal camera: Topcon TRC-50DX. Frequency: Once before laser photocoagulation, once immediately after laser induction, once before administration (day -2), once on day 14, and 28 after the first administration, for a total of 5 times.

[0157] FP image acquisition protocol: The animal's forehead was supported against a forehead support. Before capturing the image, the working distance was adjusted and focus was achieved. The acquisition of a central macular image was confirmed. The animal was managed according to the relevant SOPs during the examination.

[0158]

[0159] Fluorescein fundus angiography (FFA)

[0160] Retinal camera: Topcon TRC-50DX.

[0161] Frequency: 1 time before laser photocoagulation, 1 time before administration (-2 days), 1 time on the 14th day, and 28 days after the first administration, a total of 4 times.

[0162] FFA imaging protocol: After FP imaging, FFA was performed. After intravenous infusion of 10% sodium fluorescein (Alcon Laboratories, USA) at a dose of 0.075 mL / kg, rapid posterior polar photographs were taken of the right eye. Fluorescein leakage related to the CNV lesion was monitored using the left eye at serial time points (1, 5, and 10 minutes).

[0163] To quantify the area of ​​CNV leakage, the total area of ​​hyperfluorescence per eye was measured on late FFA images using Image J software (version 1.52a, Wayne Rasband, National Institutes of Health, USA). Images from the later stage (10 min) were also used to calculate the amount and grade of CNV lesions (Table 12).

[0164] Lesion Grade DefinitionGrade INo hyperfluorescenceGrade IIHyperfluorescence without leakageGrade IIIEarly hyperfluorescence or slight penetration and delayed leakageGrade IVBright hyperfluorescence penetration that leaks beyond the burn site borders

[0165] Optical coherence tomography (OCT): Heidelberg Spectralis OCT plus.

[0166] Frequency: 4 times in total: once before laser photocoagulation, once before administration (-2 days), and once on days 14 and 28 after the first administration.

[0167] OCT image acquisition protocol: After FP and FFA images were taken, OCT scans were performed. Efforts were made to maintain uniformly illuminated and well-focused fundus images and high-quality OCT images. The monitor screen was observed to ensure that the macula was in focus, and a high-speed macular scan process was applied to identify the monkey's eyes. Retinal thickness was automatically measured using software embedded in the Heidelberg OCT by measuring the distance between the internal limiting membrane and Bruch's membrane. Two membrane lines were manually located. The location with the greatest thickness around the class III or IV point was selected for measurement. The Heidelberg OCT has unique technical characteristics that ensure longitudinal measurements of retinal thickness at the same location.

[0168]

[0169] data analysis

[0170] Data analysis consisted of the total area of ​​fluorescein leakage spots, the number of fluorescein leakage spots, and retinal thickness at each registered photocoagulation site. Animal observations, fundus photography, and slit-lamp examinations were recorded and summarized in the report. The mean and standard deviation of the percent change in the total area of ​​fluorescein leakage spots and the percent change in retinal thickness were calculated. Microsoft Office Excel 2013 and SPSS Statistics 13.0 were used for data processing and statistical analysis.

[0171]

[0172] Results; SCAI-005 (Axitinib) monkey efficacy trial

[0173] Relative leakage area, change in leakage point

[0174] - Leakage area of ​​grade 3 / 4 CNV lesions (mm 2 , Mean±SD)

[0175]

[0176]

[0177] - Percent change of leakage area (%, Mean±SD)

[0178]

[0179]

[0180] - Amount of grade 3 / 4 leakage spots

[0181]

[0182] After inducing CNV damage, the SCAI-005 composition of Example 15 was injected. Measurements taken 14 and 28 days later confirmed that the leakage area and the number of grades 3 and 4 (leakage spots) were statistically significantly reduced compared to the control group. (Fig. 1)

[0183]

[0184] Retinal thickness (OCT) changes

[0185] - Average retinal thickness of grade 3 / 4 CNV Lesions (μm, Mean±SD)

[0186]

[0187]

[0188] - Percent change of retinal thickness (%, Mean±SD)

[0189]

[0190] After inducing CNV damage, the SCAI-005 composition of Example 15 was injected, and the results measured 28 days later confirmed that the thickness of the retinal lesion was statistically significantly reduced compared to the control group. (Fig. 2)

Claims

1. A molecular complex in which axitinib is physically bound; solubilizer; stabilizers; and A composition characterized by comprising an additive.

2. In paragraph 1, 상기 가용화제는 alphacyclodextrin (alpha-CD), beta-cyclodextrin (beta-CD), gamma-cyclodextrin (gamma-CD), 2- hydroxypropyl-beta-cyclodextrin (HP-beta-CD), sulfobutylether beta-cyclodextrin (SBE-beta- CD); 2-hydroxypropyl-alpha-cyclodextrin; randomly methylated beta-cyclodextrin; 2-O-methyl- beta-cyclodextrin; 2,6-di-O-methyl-beta-cyclodextrin; heptakis(2,3,6-tri-O-methyl)-beta- cyclodextrin; carboxymethyl-beta-cyclodextrin; carboxyethyl-beta-cyclodextrin; hydroxyethyl- beta-cyclodextrin; maltosyl-beta-cyclodextrin; 3,6-(N,N,N-trimethylammonium)propyl-beta- cyclodextrin; acetyl-beta-cyclodextrin; 2,6-di-O-methyl-gamma-cyclodextrin; 2-hydroxypropyl- gamma-cyclodextrin; or sulfobutylether gamma-cyclodextrin;optionally, a composition characterized in that the cyclodextrin comprises at least one selected from the group consisting of alpha-cyclodextrin (alpha-CD), beta-cyclodextrin (beta-CD), gamma-cyclodextrin (gamma-CD), 2-hydroxypropyl-beta-cyclodextrin (HP-beta-CD), or sulfobutylated-beta-cyclodextrin (SBE-beta-CD) and cyclodextrin derivatives.; 3. In paragraph 1, A composition characterized in that the stabilizer comprises at least one selected from the group consisting of methylcellulose (MC), hydroxymethyl cellulose (HMC), carboxymethylcellulose (CMC), hydroxypropyl methylcellulose (HPMC), hydroxyethyl cellulose (HEC) and cellulose derivatives.

4. In paragraph 1, A composition characterized in that it comprises at least one additive selected from the group consisting of a buffer, an osmotic pressure regulator, and a pH regulator.

5. In paragraph 4, A composition characterized in that the buffer comprises at least one selected from the group consisting of citric acid, phosphoric acid, boric acid, tartaric acid, acetic acid, and amino acids.

6. In paragraph 4, A composition characterized in that the osmotic pressure regulator comprises at least one selected from the group consisting of sorbitol, mannitol, dextrose, sucrose, and glycerin.

7. In paragraph 4, A composition characterized in that the pH adjusting agent is an alkalizing agent.

8. In paragraph 1, Based on the entire composition, The above molecular aggregate is 0.02 to 0.12 w / v%; The solubilizing agent is present in an amount of 2 to 12 w / v%; The stabilizer is present in an amount of 0.01 to 1.0 w / v%; A composition characterized by comprising:

9. In paragraph 4, Based on the entire composition, The buffer is 0.5 to 1.5 w / v%; The osmotic pressure regulator is 0.1 to 5 w / v%; A composition characterized by comprising:

10. In paragraph 4, A composition characterized in that the pH is adjusted to 6 to 8 using the above pH regulator.

11. In paragraph 1, A composition having an X-ray powder diffraction spectrum of the above molecular complex having X-ray diffraction peaks at diffraction angles 2θ of 24.99°±0.1° and 26.32°±0.1°.

12. In paragraph 1, A composition characterized in that the molecular assembly has a differential scanning calorimetry (DSC) profile having a glass transition at a single endothermic temperature of 220.4±2.0°C when measured under DSC conditions of 30-250°C, 99.999% N2, and a heating rate of 10°C / min.

13. In paragraph 1, A composition characterized in that the above molecular aggregate has an aspect ratio value of 0.3 to 1.

0.

14. In paragraph 1, A composition characterized in that the crystals of the above molecular aggregate have an average particle diameter of 2 to 15 μm.

15. A composition according to any one of claims 1 to 14, which is for eye drops.

16. A composition according to any one of claims 1 to 14, which is a pharmaceutical composition for preventing or treating wet macular degeneration. 17.(a) A step of dissolving the physically bound molecular complex of axitinib by adding an acid; (b) a step of preparing a first solution by adding a solubilizer to the above solution and stirring; (c) a step of preparing a second solution by solubilizing the stabilizer in water; (d) a step of mixing the first solution and the second solution; (e) a step of adding an additive including a buffer and an osmotic pressure regulator as active ingredients to the solution prepared in step (d), adjusting the pH to 6 to 8 with a pH regulator, and then adding water to mark the solution; and (f) a step of sterilizing the solution prepared in step (e); A method for producing a composition comprising:

Images