WO2022111379A1 - Axitinib intraocular implant - Google Patents

Abstract

An axitinib implant, comprising axitinib and a biodegradable polymer. The implant is stable and can achieve sustained release for at least six months, thus avoiding multiple repeated administrations, and reducing side effects. The preparation can effectively treat eye diseases such as glaucoma, cataract, retinal vein occlusion, uveitis, diabetic macular edema and age-related macular degeneration.

Description

Axitinib intraocular implant technical field

The invention belongs to the field of pharmaceutical preparations, and relates to an intraocular implant of axitinib and a preparation method.

Background technique

Eye diseases are mainly divided into two types, anterior eye diseases, including but not limited to glaucoma, cataracts, blepharospasm, conjunctivitis, conjunctivitis and corneal diseases, etc., and posterior eye diseases, including but not limited to retinal vein occlusion ( RVO), uveitis, diabetic macular edema (DME), and age-related macular degeneration (wAMD and nAMD), inflammatory or degenerative eye diseases that affect millions of people worldwide and, if not treated properly, can lead to May cause permanent visual impairment or blindness.

Systemic corticosteroid therapy has been used to treat severe uveitis. However, due to the blood-retinal barrier, only about 2% of the drugs enter the eye to play a role in systemic administration. It is a systemic adverse reaction that is generally not applicable to other chronic retinal diseases (Cheng C-K et al. (1995). Intravitreal sustained-release dexamethasone device in the treatment of experimental uveitis, Invest. Ophthalmol. Vis. Sci. 36: 442-53).

用于治疗感染眼后段的慢性非传染性眼色素层炎。 Direct ocular administration, such as intravitreal administration, provides the most targeted drug delivery, ensuring effective drug concentrations while limiting systemic exposure. Vitreous implants have been developed, such as the marketed fluocinolone vitreous implant

For the treatment of chronic non-infectious uveitis infecting the posterior segment of the eye.

Axitinib is a tyrosine kinase inhibitor with the chemical name N-methyl-2-[3-((e)2-pyridin-2-yl-vinyl)-1H-indazole-6- thio]-benzamide, having the following chemical structural formula (Compound I):

Compound I

It is currently marketed in the United States as an oral-coated tablet for the treatment of adult patients with advanced renal cell carcinoma (RCC) who have failed prior therapy with a tyrosine kinase inhibitor or cytokine. As a dual-target inhibitor of VEGF-R2 and PDGFR-B, axitinib has the desired activity to inhibit CNV formation (Giddabasappa Anand, Lalwani Kush, Norberg Rand, et al. Axitinib inhibits retinal and choroidal neovascularization in vitro and in vivo models. Experimental Eye Research, 2016.145).

Patent WO2014204791 discloses axitinib eye drops, but its drug load is small, and due to the drug barrier effect and tear dilution, etc., it prevents a sufficient amount of the drug from reaching the treatment site, requiring repeated administration, and compliance. poor. WO2017120600 discloses an ophthalmic injection of axitinib, but it does not pay attention to the stability of axitinib formulations.

The applicant of the present invention found in experiments that axitinib is highly sensitive to light, and its content varies greatly under light conditions, which is not conducive to the stability of its medicinal preparation. At the same time, its low solubility property also reduces the bioavailability of axitinib pharmaceutical preparations.

SUMMARY OF THE INVENTION

In order to overcome the shortcomings of the prior art, the purpose of the present invention is to provide a stable, safe and long-term release axitinib implant for the treatment of glaucoma, cataract, retinal vein occlusion (RVO) ), uveitis, diabetic macular edema (DME), and age-related macular degeneration (wAMD and nAMD).

The present invention includes an axitinib intraocular implant comprising axitinib and a biodegradable polymer.

In certain embodiments, the biodegradable polymer is poly(lactic-co-glycolic acid) (PLGA) copolymer, polylactide (PLA), polyglycolide (PGA), polyethylene glycol (PEG) one or more of. In certain embodiments, the biodegradable polymer is PLGA, and in one embodiment, the biodegradable polymer is ester terminated PLGA.

In certain embodiments, the ester-terminated PLGA has a molecular weight of 15,000-100,000. In certain embodiments, the ester-terminated PLGA has a molecular weight of 15,000-70,000. In a preferred embodiment, the molecular weight of the ester-terminated PLGA is 70,000. In a preferred embodiment, the molecular weight of the ester-terminated PLGA is 15,000.

In certain embodiments, the ratio of glycolic acid to lactic acid monomers in the PLGA is from 0:100 to 100:0, and in certain embodiments, the ratio of glycolic acid to lactic acid monomers is from 15:85 to 85:15 , in certain embodiments, the ratio of glycolic acid and lactic acid monomers is 50:50.

In certain embodiments, the implant comprises 20%-90% by weight axitinib, in certain embodiments, the implant comprises 30%-80% by weight axitinib, in In certain embodiments, the implant comprises 30%-70% by weight of axitinib, in certain embodiments, the implant comprises 40%-70% by weight of axitinib, in certain In some embodiments, the implant comprises 45%-70% by weight of axitinib, in certain embodiments, the implant comprises 40%-50% by weight of axitinib, in certain embodiments In regimens, the implant comprises 45%-50% by weight of axitinib, and in certain embodiments, the implant comprises 50%-70% by weight of axitinib. In certain embodiments, the implant comprises 30%-80% axitinib and 20%-70% ester-terminated PLGA by weight. In certain embodiments, the intraocular implant comprises 50%-70% axitinib and 30%-50% ester terminated PLGA by weight. In certain embodiments, the intraocular implant comprises 40%-70% axitinib and 30%-50% ester-terminated PLGA by weight. In certain embodiments, the intraocular implant comprises 40%-50% axitinib and 40%-50% ester terminated PLGA by weight. In certain embodiments, the intraocular implant comprises 70% by weight of axitinib and 30% by weight of ester-terminated PLGA. In certain embodiments, the intraocular implant comprises by weight 50% axitinib and 40% ester terminated PLGA. In certain embodiments, the intraocular implant comprises by weight 50% axitinib and 45% ester terminated PLGA. In certain embodiments, the intraocular implant comprises by weight 50% axitinib and 50% ester terminated PLGA. In certain embodiments, the intraocular implant comprises 50% by weight axitinib and 50% ester-terminated PLGA, wherein the ester-terminated PLGA has a molecular weight of 70,000. In certain embodiments, the intraocular implant comprises 50% axitinib by weight and 50% ester-terminated PLGA, wherein the ester-terminated PLGA has a molecular weight of 15,000.

In certain embodiments, axitinib is included in the implant in a weight of 200 micrograms to 1.5 mg, in certain embodiments, axitinib is included in the implant in a weight of 200 micrograms to 1 mg, In certain embodiments, axitinib is included in the implant in a weight of 1 mg to 1.5 mg. In certain embodiments, the intraocular implant weighs 200 micrograms, 400 micrograms, 500 micrograms, 600 micrograms, 700 micrograms, 1 mg, or 150 mg.

In certain embodiments, release modifiers are included in the implant including hydroxypropyl methylcellulose, hyaluronic acid and its sodium salts, poloxamers, polyethers, polyvinyl alcohol, one or more of polyvinylpyrrolidone, lactose, polyethylene glycol, mannitol, glucose, maltose, sodium chloride, potassium chloride, magnesium carbonate, sodium bicarbonate, potassium bicarbonate, or sucrose, in certain implementations In the scheme, the release modifier includes one or more of polyethylene glycol, mannitol, sodium chloride, and polyvinylpyrrolidone. In certain embodiments, the release modifier includes one or more of mannitol, sodium chloride, polyvinylpyrrolidone.

In certain embodiments, the release modifier is present in an amount of 0% to 10% by weight, in certain embodiments, the release modifier is present in an amount of 2.5% to 10% by weight, in certain embodiments In embodiments, the release modifier is present in an amount of 5% to 10% by weight, in certain embodiments, the release modifier is present in an amount of 0% by weight, and in certain embodiments, the release modifier The agent content is 10% by weight.

In certain embodiments, the intraocular implant comprises: by weight 50% axitinib, 40% ester-terminated PLGA, and 10% mannitol, the ester-terminated PLGA having a molecular weight of 15,000 or 70,000.

In certain embodiments, the intraocular implant comprises: 50% by weight axitinib, 40% ester-terminated PLGA, and 10% polyvinylpyrrolidone, the ester-terminated PLGA molecular weight 15,000 or 70,000.

In certain embodiments, the intraocular implant comprises: 50% by weight axitinib, 45% ester-terminated PLGA, and 5% sodium chloride, the ester-terminated PLGA molecular weight 15,000 or 70,000.

In certain embodiments, intraocular implants are used for vitreous administration.

The present invention further provides a preparation method for preparing the above-mentioned intraocular implant. In certain embodiments, the preparation method adopts a hot melt extrusion method, in certain embodiments, the preparation method adopts a single extrusion or double extrusion method, and in certain embodiments, a single extrusion method is adopted. Secondary extrusion method.

In one embodiment, the preparation method may include the specific steps of: pulverizing axitinib, physically mixing with polymers and/or other optional excipients (eg, release modifiers, etc.) The screw hot-melt extruder extrudes, gradually heats up, and the temperature setting range is between 90°C and 180°C.

In one embodiment, the preparation method may include the following specific steps: separately pulverizing axitinib and the polymer, physically mixing with other optional excipients (such as release modifiers, etc.), etc., adding the above mixture to the twin-screw The hot melt extruder extrudes, gradually heats up, and the temperature setting range is between 90°C and 180°C.

In one embodiment, the preparation method includes the steps of: physically mixing axitinib with PLGA and/or release modifier, adding the above mixture into a twin-screw hot-melt extruder, gradually increasing the temperature, and the temperature is set in the range of 90 Between ℃ and 180 ℃, extrude, crush the rod-shaped object formed by the first extrusion, add it to the twin-screw extruder again, and perform the second extrusion molding. After the extrusion is completed, the extruded material is cut to desired length.

The present invention provides the use of the above implant in the manufacture of a medicament for ocular diseases, examples of which include but are not limited to glaucoma, cataract, retinal vein occlusion, uveitis, diabetic macular edema and age-related macular degeneration.

The present invention provides a stable, safe and sustained-release axitinib implant for at least 6 months, which can effectively treat diseases including but not limited to glaucoma, cataract, retinal vein occlusion (RVO), uveitis, diabetes For various eye diseases such as macular edema (DME) and age-related macular degeneration (wAMD and nAMD), multiple repeated administrations are avoided, the formation of side effects is reduced, the cost of medication is reduced, and the production, storage, and transportation of pharmaceutical preparations are satisfied. It has a high degree of stability required in the application, and has a good application prospect to meet the unmet clinical needs.

Description of drawings

Figure 1 shows the in vitro release profiles of formulations 1 to 3

Fig. 2 is the intraocular fluorescence angiography of the monkey eye of prescription 22 (A is before modeling, B is after modeling, C is four weeks after administration)

Detailed ways

Various aspects will now be described more fully below. Such aspects, however, may be embodied in many different forms and should not be construed as limited to the embodiments set forth herein; rather, these embodiments are provided so that this disclosure will be thorough and complete, and will fully convey to those skilled in the art show its scope.

definition

The following terms used herein have the following meanings:

"Drug" means any substance used to treat eye conditions.

"Intraocular" means the entire ocular region including but not limited to the anterior chamber, posterior chamber, vitreous cavity, choroid, periocular, ocular surface, suprachoroidal space, conjunctiva, subconjunctival space, cornea, intracorneal space, cornea External space, sclera, etc.

"Molecular weight" refers to the general term for relative molecular mass, that is, the sum of the atomic weights of the atoms that make up the molecule.

"Biodegradable polymer" refers to a polymer that can be converted in vivo to non-toxic degradation products over time, wherein degradation of the polymer over time is required to obtain the drug release kinetics according to the present invention.

"Ester-terminated" means having an ester bond at the end of the polymer, and typical ester-terminated groups include, but are not limited to, alkyl esters and aromatic esters.

"Ocular disease" refers to a disease, disorder or disorder affecting or involving the eye or an area or area of the eye, such as retinal disease. The eye includes the eyeball and the tissues and fluids that make up the eyeball, periocular muscles (eg, the oblique and rectus muscles), and the portion of the optic nerve within or adjacent to the eyeball.

"Implant" refers to a sterile preparation made of drugs and excipients for implantation into the body, including subcutaneous implantation, vitreous implantation, and contraceptive ring implantation. The term "implant" is used interchangeably with "implant"

Whenever the term "at least" precedes the first value of a series of two or more values, the term "at least" applies to each value in the series.

The present invention provides axitinib implants and methods for treating ocular disorders. Typically, the implant continues to release axitinib to the ocular area for a period of time including, but not limited to, twelve months, nine months, six months, three months, or less than three months.

Axitinib intraocular implant

Implants of the present invention comprise axitinib dispersed in a biodegradable polymer for direct placement in the eye. In certain embodiments, axitinib constitutes 20-90% by weight of the implant, in certain embodiments, axitinib constitutes 30-80% by weight of the implant, in one embodiment, Axitinib accounts for 50-70% by weight of the implant, in one embodiment axitinib accounts for 50% by weight of the implant, in one embodiment axitinib accounts for 50% by weight of the implant 70%. In certain embodiments, axitinib is included in the implant in a weight of 200 micrograms to 1.5 mg, in certain embodiments, axitinib is included in the implant in a weight of 200 micrograms to 1 mg, In certain embodiments, axitinib is included in the implant in a weight of 1 microgram to 1.5 milligrams. In certain embodiments, axitinib is included in the implant in a weight of 1.5 mg, and in certain embodiments, axitinib is included in the implant in a weight of 1 mg. In certain embodiments, axitinib is included in the implant in a weight of 200 micrograms. In certain embodiments, axitinib is included in the implant in a weight of 400 micrograms. In certain embodiments, axitinib is included in the implant in a weight of 500 micrograms. In certain embodiments, axitinib is included in the implant in a weight of 600 micrograms. In certain embodiments, axitinib is included in the implant in a weight of 700 micrograms, and in certain embodiments, axitinib is included in the implant in a weight of 750 micrograms.

In certain embodiments, axitinib can be uniformly dispersed in the biodegradable matrix of the pharmaceutical composition. The choice of the biodegradable polymer matrix used can vary depending on the desired release kinetics, patient tolerance, the nature of the disease to be treated, and the like. Considered polymer properties include, but are not limited to, biocompatibility and biodegradability at the site of administration, compatibility with axitinib, and processing temperature. In certain embodiments, the biodegradable polymer matrix is 20%-70% by weight of the implant, in one embodiment, the biodegradable polymer matrix is generally 30%-50% by weight of the implant, In one embodiment the biodegradable polymer matrix is typically 50% by weight of the implant, in one embodiment the biodegradable polymer matrix is typically 30% by weight of the implant, in one embodiment, The biodegradable polymer matrix is typically 40% by weight of the implant, and in one embodiment the biodegradable polymer matrix is typically 45% by weight of the implant. .

Useful biodegradable polymer matrices include, but are not limited to, polymers made from monomers such as organic esters or organic ethers, which upon degradation produce physiologically acceptable degradation products. Anhydrides, amides or orthoesters can also be used to polymerize themselves or to polymerize with other monomers. The polymers are usually condensation polymers. The polymers can be crosslinked or non-crosslinked.

For most polymers, in addition to carbon and hydrogen, oxygen and nitrogen are included, especially oxygen, which may be present in the form of oxygen-containing groups such as hydroxyl, carboxyl, ether, carbonyl, ester, and the like. Nitrogen can exist in the form of amide, amino and the like. A description of a list of examples of biodegradable polymers that can be used can be found in Heller, Biodegradable polymers in controlled drug delivery, In: "CRC critical reviews in therapeutic drug carrier systems", Vol. 1, CRC press, Boca Raton, FL (1987) .

In some embodiments, the biodegradable polymer comprises at least one of the following: poly(lactic-co-glycolic acid) (PLGA), polylactide (PLA), polyglycolide (PGA), polyethylene glycol Alcohol (PEG), D-lactide, D,L-lactide, L-lactide, D,L-lactide-co-ε-caprolactone, L-lactide-co-ε-caprolactone, D,L-lactide-co-glycolide-co-ε-caprolactone, poly(D,L-lactide-co-caprolactone), poly(L-lactide-co-caprolactone) , poly(D-lactide-co-caprolactone), poly(D,L-lactide), poly(D-lactide), poly(L-lactide), poly(esteramide), or a combination thereof. In some embodiments, the biodegradable polymer comprises poly(lactide-co-glycolide) (PLGA).

Copolymers of glycolic acid and lactic acid are of particular interest, and the percentages of glycolic acid and lactic acid monomers in the PLGA may be 0-100%, 15-85%, 25-75%, or 35-65%. In a preferred embodiment, PLGA is used in a 50:50 ratio of glycolic acid to lactic acid monomers.

On the other hand, the inventors found that the use of ester-terminated PLGA as a biodegradable polymer matrix can enhance the stability of axitinib pharmaceutical compositions, including but not limited to stability under high temperature and high humidity, light stability, long-term stability, etc. Ester-terminated PLGAs have ester linkages at the polymer ends, and typical ester-terminated groups include, but are not limited to, alkyl esters and aromatic esters. The molecular weight of the ester-terminated PLGA is between 15,000 and 100,000. In a variation, the molecular weight of the ester-terminated PLGA is between 15,000 and 70,000. In one embodiment, the molecular weight of the ester-terminated PLGA is 70,000.

In certain embodiments, axitinib may be pulverized into granules or micropowders, at least 90% of the axitinib granules or micropowders having a diameter of less than or equal to 20 microns, and in a variation, the at least 90% % Axitinib particles or micropowders are less than or equal to 15 microns in diameter.

The biodegradable polymers of the present invention may optionally be comminuted into granules or micropowders, with respect to the size of the polymer granules or micropowders, in a variation, at least 90% of the polymer micropowders or particles are 10-100 μm in diameter, in a In variations, at least 90% of the polymer micropowders or particles are 10-50 μm in diameter, and in a variation, at least 90% of the polymer micropowders or particles are 20-40 μm in diameter.

In certain embodiments, the implants of the present invention may also optionally and/or preferably contain other excipients, eg, release modifiers may be used. The release modifier can be used to accelerate the release of axitinib while maintaining a smooth release profile of axitinib. In one embodiment, the release modifier is one or more polysaccharides, such as cellulose-based materials, including: Hydroxypropyl methylcellulose, hyaluronic acid, poloxamers, polyethers such as polyethylene glycol, polyvinyl alcohol, polyvinylpyrrolidone, and mixtures thereof. In one embodiment, the release modifier is a pore former and/or a stability enhancer, optionally lactose, mannitol, glucose, maltose, sodium chloride, potassium chloride, magnesium carbonate, sodium bicarbonate, Potassium bicarbonate or sucrose etc. In another embodiment, the release modifier is a mixture of two or more of the aforementioned modifiers. In one embodiment of the invention, the pharmaceutical composition does not contain any release modifier. In one embodiment of the present invention, the release modifier is sodium chloride, in one embodiment of the present invention, the release modifier is polyvinylpyrrolidone, in one embodiment of the present invention, the release modifier is Mannitol. In certain embodiments of the present invention, the release modifier is present in an amount of 0% to 10% by weight, in certain embodiments of the present invention, the release modifier is present in an amount of 2.5% to 10% by weight, In certain embodiments of the present invention, the release modifier is present in an amount of 5% to 10% by weight, in certain embodiments, the release modifier is present in an amount of 0% by weight, in certain embodiments , the release modifier is present in an amount of 10% by weight, in certain embodiments, the release modifier is present in an amount of 5% by weight, and in certain embodiments, the release modifier is present in an amount of 2.5% by weight.

The intraocular implants herein are generally solid and can be prepared in the shape of particles, sheets, blocks, films, fibers, rods, discs, etc., or can be of any size or shape compatible with the chosen implantation site, so long as The implant has the desired release kinetics and is capable of delivering an amount of a drug that can treat ocular diseases. In certain embodiments, according to the dosage of the drug contained in the implant, the duration of continuous administration and the clinical administration mode, the implant of the present invention preferably has a long cylindrical or thin rod-like appearance, with a length of 0.3 mm to 10 mm. mm, diameter 0.05mm-1mm, total weight 100-5000 micrograms, usually 500-1500 micrograms. In certain embodiments, the implant is 5 millimeters in length, 0.5 millimeters in diameter, and has a total weight of 1000 micrograms. In certain embodiments, the implant is 0.5 millimeters in diameter, 5-7 millimeters in length, and weighs about 1.0-1.5 milligrams.

Preparation

The present invention provides a preparation method for preparing an implant comprising the above-mentioned axitinib pharmaceutical composition, and the preparation method can be carried out by a hot melt extrusion method, for example, using a piston extruder, a single screw hot melt extrusion extruder or twin-screw hot-melt extruder to uniformly disperse and distribute axitinib in the biodegradable polymer. In certain embodiments, the implant is prepared by a single extrusion or double extrusion, and in preferred embodiments, a single extrusion is used. Typically, the hot melt extrusion process is used at a temperature of about 25°C to 180°C, in some embodiments, 90°C to 180°C, in some embodiments, 130°C to 180°C, and In certain embodiments, the temperature is 150 to 180°C. In one embodiment, the single extrusion method may include the steps of: pulverizing axitinib, physically mixing it with a polymer and/or other optional agents (such as release modifiers, etc.), Add the above mixture into the twin-screw hot melt extruder, gradually heat up, set the temperature between 90°C and 180°C, and control the speed between 5RPM and 30RPM, subject to the mixture being melted and fully mixed, extruding, extruding After completion the extrudate is cut to the desired length, in some embodiments 5 mm in length.

In another embodiment, the preparation method may include the steps of: pulverizing axitinib and the polymer into granules or micropowders, respectively, and mixing the above granules or micropowders and/or other optional agents (such as release modifiers, etc.), etc. Physical mixing, add the above mixture into the twin-screw hot-melt extruder, gradually heat up, set the temperature between 90°C and 180°C, and control the speed between 5RPM and 30RPM, subject to the mixture being melted and fully mixed, extruding After pressing, the extrudate is cut to the desired length, in some embodiments 5 mm in length.

In another embodiment, the preparation method may include the steps of: physically mixing axitinib with PLGA and/or release modifier, adding the above mixture into a twin-screw hot-melt extruder, gradually increasing the temperature, and setting the temperature within the range Between 90℃-180℃, control the speed between 5RPM and 30RPM, subject to the mixture being melted and fully mixed, extrude, pulverize the rod-shaped object extruded for the first time, and then add the twin-screw hot melt extrusion Out of the machine, a second extrusion molding is performed, and after the extrusion is completed, the extrudate is cut to the desired length, in some embodiments, the length is 5 mm.

Implantation method

Biodegradable implants can be placed in the eye by a variety of methods, including placement through forceps, syringes, trocars, or other types of drug delivery devices after making an incision in the iris. In some cases, forceps, syringes, trocars, or other types of drug delivery devices can be used without an incision. In a preferred variation, one or more implants can be placed into the eye using a hand-held drug delivery device.

The implantation method typically involves first entering a needle into a target area in the ocular region, and after entering the target area, eg, the vitreous cavity, pushing a handle on the hand-held drug delivery device to cause the pusher to advance the plunger. As the plunger advances, it pushes the implant into the target area.

application

The implants of the present invention are useful in the treatment of ocular diseases, examples of which include, but are not limited to, glaucoma, cataracts, retinal vein occlusion, uveitis, diabetic macular edema, and age-related macular degeneration.

Detailed ways

The present invention is further described in detail by the following examples, which cannot be used to limit the protection scope of the present invention.

In the following specific embodiment, the medicines, reagents and instruments used are as follows:

equipment:

Drugs, excipients and reagents:

name Model/Specification company axitinib Pharmaceutical grade Zhejiang Jiuzhou Pharmaceutical Co., Ltd. PLGA RG502 Evonik Industries PLGA RG502H Evonik Industries PLGA RG505 Evonik Industries PLGA RG858S Evonik Industries PLA R205S Evonik Industries PEG 3350 Clariant methanol HPLC grade Chengdu Kelon Chemical Co., Ltd. Acetonitrile HPLC grade Chengdu Kelon Chemical Co., Ltd. Nacl AR level Sigma SDS AR level Sigma Mannitol 160C, pharmaceutical grade Roquette PVP VA64, pharmaceutical grade BASF

In vitro release experiments

Preparation method of release medium: release medium: 0.9% NaCl+0.3% SDS;

Weigh 9 g of sodium chloride to 1000 ml of degassed water, dissolve it, add 3 g of sodium dodecyl sulfate, and sonicate for 30 minutes.

Set the temperature of the constant temperature oscillator to 37.0±0.5°C, the rotation speed to 150RPM, and the operation mode: gyration; use a 10ml vial to add the prepared 0.9%NaCl+0.3%SDS solution as the release medium, and add 4ml to each bottle; The sample of the release medium is added to a constant temperature oscillator for temperature balance. After the temperature of the measured liquid reaches 37°C, a cut implant preparation with a diameter of 0.5mm and a length of about 5-7mm is added, and shaken to make it sink to the liquid level Put into the thermostatic oscillator, run the sample in insulation, measure the concentration of API in the medicinal liquid by high-performance liquid chromatography (chromatographic conditions are as follows) sampling at a predetermined time, wherein the sampling method is to pipet 3ml of 4ml of release liquid in the sample bottle , and replaced with 3ml of freshly prepared 0.9%NaCl+0.3%SDS solution, put it back into the equipment for investigation; and calculated the single release amount and cumulative release amount of the sample, and formed the release behavior measurement result.

Stability test: The samples were exposed for 5 days, 10 days or 30 days under the conditions of high temperature (60°C), high humidity (92.5%RH), and light (4500lx±500lx), and try to avoid other conditions. of cross effects.

Content detection method: Determination according to high performance liquid chromatography

Chromatographic conditions and system suitability experiment: use octadecylsilane-bonded silica gel as filler, acetonitrile-water (50:50) as mobile phase, flow rate 1.0ml/min, detection wavelength 330nm, column temperature: 35℃ ,

Example 1

Table 1 Formulation composition

Preparation Process

Add axitinib to the jet mill to pulverize to the target particle size D90≤15 microns, then physically mix the axitinib powder with the excipients, add it to the twin-screw hot melt extruder at a certain rate, and use a 0.5mm diameter die. Mouth, heating, gradually heating up, the temperature setting range is between 90 ℃-180 ℃, heating to prepare extrudate, it is advisable to extrude smoothly. The extruded samples were stretched using a crawler-type adaptor, and the speed was controlled between 5 RPM and 30 RPM to match the speed at which the extruded rods were collected smoothly. Each of the above prescriptions is prepared in a way that the total amount of the prescription is 7g, and is prepared according to the proportion of the above prescription list, and the extruded preparation sample is controlled to be an implant preparation with a diameter of 0.5mm, a length of 5mm, and a weight of about 1.0mg.

Release Degree Test Rub:

The release results are shown in Figure 1.

Stability check:

Stability research was carried out on prescription 1, prescription 2, and prescription 3, and they were exposed for 5-10 days under the conditions of high temperature (60°C), high humidity (92.5% RH), and light (4500lx±500lx), respectively, and Try to avoid cross-effects caused by other conditions. The stability results are shown in Table 2-Table 4:

Table 2 5-day stability data at high temperature

Table 3 5-day stability data under high humidity

Table 4 5-day stability data under light

Embodiment 2

Table 5 Formulation composition

Preparation Process

Add API into jet mill to pulverize, then mix API powder and auxiliary materials physically, add into twin-screw hot-melt extruder at a certain rate, use 0.5mm diameter circular die, and temperature setting range is 150℃-180℃ In between, the extrudate is prepared by heating, and it is advisable to extrude smoothly. The extruded samples were stretched using a crawler-type adaptor, and the speed was controlled between 5 RPM and 30 RPM to match the speed at which the extruded rods were collected smoothly. The prescription is prepared in a way that the total amount of the prescription is 6g, and is prepared according to the proportion of the above prescription list. The extruded preparation sample is controlled to have a diameter of 0.5mm, a length of 5-6mm, and a weight of about 1.0-1.5mg. The unit implant preparation.

Stability check:

The stability study was carried out on prescription 3 and prescription 4, and they were exposed to light (4500lx ± 500lx) for 5 days, and the cross-influence caused by other conditions was avoided as much as possible. The stability results are shown in Table 6:

Table 6 5-day stability data under light

Embodiment 3

Table 7 Formulation composition

Preparation Process

Add API into jet mill to pulverize, then physically mix API powder and auxiliary materials, and add to twin-screw hot-melt extruder at a certain rate, using a 0.5mm diameter circular die, and the temperature setting range is 90℃-180℃ In between, the extrudate is prepared by heating, and it is advisable to extrude smoothly. The extruded samples were stretched using a crawler-type adaptor, and the speed was controlled between 5 RPM and 30 RPM to match the speed at which the extruded rods were collected smoothly. The prescription is prepared in a way that the total amount of the prescription is 6g, and is prepared according to the proportion of the above prescription list. The extruded preparation sample is controlled to have a diameter of 0.5mm, a length of 5-6mm, and a weight of about 1.0-1.5mg. The unit implant preparation.

Stability check:

Stability study was carried out on prescriptions 6-8, and they were placed naked for 10 days under three conditions of light (4500lx ± 500lx), and the cross-influence brought by other conditions was avoided as much as possible. The stability results are shown in Table 8:

Table 8 10-day stability data under light

Embodiment 4

Table 9 Formulation composition

Preparation Process

Add the API into the jet mill to pulverize, then physically mix the API powder with the auxiliary materials, and add it to the twin-screw hot melt extruder at a certain rate, using a 0.5mm diameter circular die, and the temperature setting range is 130℃-180℃ In between, the extrudate is prepared by heating, and it is advisable to extrude smoothly. The extruded samples were stretched using a crawler-type adaptor, and the speed was controlled between 5 RPM and 30 RPM to match the speed at which the extruded rods were collected smoothly. The prescription is prepared in a way that the total amount of the prescription is 6g, and is prepared according to the proportion of the above prescription list. The extruded preparation sample is controlled to have a diameter of 0.5mm, a length of 5-6mm, and a weight of about 1.0-1.5mg. The unit implant preparation.

Stability check:

Stability research was carried out on prescription 9-11, which was exposed to light (4500lx±500lx) for 10 days under three conditions, and the cross-influence caused by other conditions was avoided as much as possible. The stability results are shown in Table 10:

Table 10 10-day stability data under light

Embodiment 5

Table 11 Formulation composition

Preparation Process

Add API and auxiliary materials to the jet mill for pulverization, and then physically mix the API powder and auxiliary material powder, and add them to the twin-screw hot melt extruder at a certain rate, using a 0.5mm diameter circular die, and the temperature setting range is 150 ℃ Between -180 ℃, heating to prepare extrudate, it is advisable to extrude smoothly. The extruded samples were stretched using a crawler-type adaptor, and the speed was controlled between 5 RPM and 30 RPM to match the speed at which the extruded rods were collected smoothly. The prescription is prepared in a way that the total amount of the prescription is 6g, and is prepared according to the proportion of the above prescription list. The extruded preparation sample is controlled to have a diameter of 0.5mm, a length of 5-6mm, and a weight of about 1.0-1.5mg. The unit implant preparation.

Stability check:

Stability study was carried out on formulations 12-13, which were exposed to light (4500lx ± 500lx) for 10 days under three conditions, and the cross-influence caused by other conditions was avoided as much as possible. The stability results are shown in Table 12:

Table 12 10-day stability data under light

Embodiment 6

Table 13 Formulations of preparations

component prescription 14 axitinib 70 PLGA RG858S 30

Preparation Process

Add axitinib to the jet mill to pulverize to the target particle size D90≤15 microns, then physically mix the axitinib powder with the excipients, add it to the twin-screw hot melt extruder at a certain rate, and use a 0.5mm diameter die. Mouth, heating, gradually heating up, the temperature setting range is between 90 ℃-180 ℃, heating to prepare extrudate, it is advisable to extrude smoothly. The extruded samples were stretched using a crawler-type adaptor, and the speed was controlled between 5 RPM and 30 RPM to match the speed at which the extruded rods were collected smoothly. The extrudate was collected as a pale yellow rod. The above prescription is prepared in a way that the total amount of the prescription is 7g, according to the proportion of the above prescription list, and the extruded preparation sample is controlled to be an implant preparation with a diameter of 0.5mm, a length of 5mm, and a weight of about 1.0mg.

Embodiment 7

Table 14 Formulation formulation

component prescription 15 axitinib 30

PLA R205S 70

Preparation Process

Add axitinib to the jet mill to pulverize to the target particle size D90≤15 microns, then physically mix the axitinib powder with the excipients, add it to the twin-screw hot melt extruder at a certain rate, and use a 0.5mm diameter die. Mouth, heating, gradually heating up, the temperature setting range is between 90 ℃-180 ℃, heating to prepare extrudate, it is advisable to extrude smoothly. The extruded samples were stretched using a crawler-type adaptor, and the speed was controlled between 5 RPM and 30 RPM to match the speed at which the extruded rods were collected smoothly. The extrudate was collected as a pale yellow rod. The above recipe is prepared in a way that the total amount of the recipe is 7g, and is prepared according to the proportion of the above prescription list, and the extruded preparation sample is controlled to be an implant preparation with a diameter of 0.5mm, a length of 5mm, and a weight of 1.0mg.

Embodiment 8

Table 15 Formulation formulation

component prescription 16 axitinib 50 PLGA RG505 50

Preparation Process

Physically mix axitinib powder (particle size D90≤76.6μm) with excipients, add it to a twin-screw hot-melt extruder at a certain rate, use a 0.5mm diameter die, heat, and gradually increase the temperature. The temperature setting range is Between 90°C and 180°C, the extrudate is prepared by heating, which is suitable for smooth extrusion. The extruded samples were stretched using a crawler-type adaptor, and the speed was controlled between 5 RPM and 30 RPM to match the speed at which the extruded rods were collected smoothly. The extrudate was collected as a pale yellow rod. The above recipe is prepared in a way that the total amount of the recipe is 7g, and is prepared according to the proportion of the above prescription list. The extruded preparation sample is controlled to be an implant preparation with a diameter of 0.5mm, a length of 5mm, and a weight of 1.0mg.

Embodiment 9

Table 16 Formulation formulation

component prescription 17 prescription 18 axitinib 50 50 PLGA RG502 50 / PLGA RG505 / 50

Preparation Process

Add axitinib to the jet mill and pulverize it to the target particle size D90≤15 microns, physically mix the axitinib powder with the auxiliary powder, and add it to the twin-screw hot melt extruder at a certain rate, using a 0.5mm diameter die. Mouth, heating, gradually heating up, the temperature setting range is between 90 ℃-180 ℃, heating to prepare extrudate, it is advisable to extrude smoothly. The extruded samples were stretched using a crawler-type adaptor, and the speed was controlled between 5 RPM and 30 RPM to match the speed at which the extruded rods were collected smoothly. The rod-shaped object extruded for the first time is ground and pulverized, and then added to the twin-screw hot-melt extruder again for the second extrusion molding, and the extrudate is collected. The extrudate was obtained as a pale yellow rod. Each of the above recipes is prepared in a way that the total amount of the recipe is 7g, and is prepared according to the proportions listed in the above recipes. The extruded preparation sample is controlled to be an implant preparation with a diameter of 0.5mm, a length of 5mm, and a weight of 1.0mg.

Example 10 In vivo release experiments in monkey eyes

Sample: Axitinib implants, prepared according to Recipe 19, Recipe 20 (Table 17).

Preparation process: add axitinib to the jet mill to pulverize to the target particle size D90≤15 microns, then physically mix the axitinib powder with the auxiliary materials, and add it to the twin-screw hot melt extruder at a certain rate, using 0.5 mm diameter die, heating, gradually heating up, the temperature setting range is between 90 ℃-180 ℃, heating to prepare extrudate, it is advisable to extrude smoothly. The extruded samples were stretched using a crawler-type adaptor, and the speed was controlled between 5 RPM and 30 RPM to match the speed at which the extruded rods were collected smoothly. Cut the extrudate to the desired length.

Table 17

prescription PLGA model Drug loading(%) 19 RG502 70 20 RG505 70

Animals: 2 cynomolgus monkeys aged 2.5 to 6, purchased from Chengdu Huaxi Haiqi Pharmaceutical Technology Co., Ltd. The average body weight at the time of purchase and modeling was 2.5-6.0 kg, and the individual body weight was within the range of mean ± 20%.

Animal groups: 1 monkey was given prescription 19 as prescription 19 group, and another monkey was given prescription 20 as prescription 20 group.

Experimental drugs: 2 samples of prescription 19 (the size of each preparation is 0.5mm*7mm, the total weight is about 1.42mg); 2 samples of prescription 20 (the size of each preparation is 0.5mm*7mm, the total weight is about 1.45mg)

Dosing process:

1) Anesthetize a monkey after immobilization, and after observing that there is no obvious physiological trunk activity, use povidone-iodine solution to disinfect the monkey's eyes;

2) The vitreous body was implanted into the monkey's left eye by filling one implant of prescription 19 with a 22G needle, and the implant was pushed into the vitreous body of the monkey, and the implant was implanted;

3) The same method was used to formulate 20 implants. The implants were implanted in the left eye of another monkey.

Sampling and testing: Prescription 19 When the monkeys were reared to 1 month, after the monkeys were killed, the intraocular vitreous humor, lens and residual drug were collected and marked respectively, and the concentration of each tissue was determined by high performance liquid phase method. In addition, 20 monkeys were prescribed and raised for 6 months, then the monkeys were killed, and the intraocular vitreous humor, lens and residual drugs were collected. The results are shown in Table 18-Table 19.

Determination method of drug in lens: Determination by high performance liquid chromatography (General Rule 0512 of Chinese Pharmacopoeia 2020), animal plasma standard with axitinib concentration of 0.2, 1, 4, 10, 14, 20ug/mL 6 concentration gradients were prepared. Axitinib content in the lens was calculated according to the following formula.

Determination method of drugs in vitreous humor: using high performance liquid chromatography (Chinese Pharmacopoeia 2020 edition general rule 0512), prepare animal plasma with axitinib concentration of 0.2, 1, 4, 10, 14, 20ug/mL 6 concentration gradients respectively Standard solution, draw a standard curve by plotting the concentration against the peak area, and calculate the axitinib content in the vitreous humor according to the following formula.

Calculation formula:

Standard curve: Y=KX+b

where:

Y: peak area of axitinib in standard solution;

X: Axitinib concentration in standard solution;

Sample A: Axitinib peak area in the sample to be tested;

V dilution volume: the dilution ratio of the vitreous humor to be tested.

Determination method of residual drug content:

High performance liquid chromatography (Chinese Pharmacopoeia 2020 edition general rule 0512) was used to determine the content of axitinib in the remaining drugs with axitinib as the reference substance, and the external standard method. Calculated as follows:

where:

_Sample A: Axitinib peak area in the sample to be tested;

C _control : control solution concentration;

对照溶液平均峰面积；

The average peak area of the control solution;

V _{dilution volume} : the dilution volume of the sample to be tested;

m _{weight of drug segment: the weight} of the drug formulation.

Table 18. 1-month sample release data of formulation 19 in monkey eyes

Table 19. Formulation 20 6-month sample release data in monkey eyes

Example 11 Pharmacodynamic Experiment

The above-mentioned prescription 20 was confirmed and verified by pharmacodynamics under the monkey CNV model, as follows:

Modeling method:

A single injection of prescription 20 implant through the eye vitreous was used to study the efficacy of laser-induced choroidal neovascularization leakage and growth in cynomolgus monkeys. , to establish an animal model similar to that of human choroidal neovascularization. Fluorescein angiography was performed before photocoagulation and at 21 and 28 days after photocoagulation to determine the condition of modeling.

Grouping method:

One cynomolgus monkey (male) was taken as the prescription 20 groups.

Application and detection methods:

Twenty-one days after photocoagulation, 20 groups of prescription models were given vitreous injection in both eyes with 20 prescriptions per eye. Before modeling, before grouping, and on the 7th, 14th, and 28th days after administration, the 20 groups of animals were subjected to fundus color photography, fluorescein fundus angiography and OCT examinations to observe the inhibition of choroidal neovascularization by the test product. Fundus examination was performed every 2 weeks after 29 days after administration, and gross autopsy was performed on the 51st day and the 204th day of the experiment.

The experimental results are summarized as follows:

(1) General situation

The 20 groups of monkeys who were prescribed were in good mental condition, normal autonomous activities, and normal appearance.

(2) AMD modeling

Before the monkey modeling, the color photography of the fundus of all eyeballs in the 20 groups showed no abnormal changes in the retina and retinal blood vessels, and no fluorescein leakage was found in the fundus angiography examination. Uniform thickness. 20 days after laser modeling (before administration), 9 laser spots around the macula were seen in the eye fundus color photography of the 20 groups of monkeys with prescription, and there were laser spots around the macula with high fluorescence, obvious fluorescein leakage and leakage. beyond the edge of the spot.

(3) Fluorescence angiography

28 days after administration of the prescription 20 groups of monkeys, the area of fluorescent spots in the left eye was reduced, and the area of fluorescein leakage was reduced by 10.821 mm ² ; the area of fluorescent spots in the right eye was reduced 28 days after administration, and the area of leakage of fluorescein decreased. The amount is 7.193mm ² . At 14 and 28 days after administration, the number of grade 4 fluorescent spots were 5 and 3, respectively, which were reduced compared with 12 before administration.

Table 20: Fluorescent spot area (mm ² )

Note: The area of fluorescein leakage is 0, which means that no fluorescein leakage is found in FFA examination.

(4) Fundus examination

On the 65th, 79th, 93rd, 107th, 121st, 135th, 149th, 163rd, 177th, 191st and 204th days of the prescription 20 groups of monkeys, the optic disc was vaguely visible in the fundus; anterior chamber cells.

To sum up, under the conditions of this experiment, the cynomolgus monkeys with laser CNV model were given Prescription 20 implant by single injection into the vitreous body of both eyes. Monkey CNV has inhibitory effect. The axitinib implant of the present application can achieve a sustained-release therapeutic effect of not less than 6 months, has stability, and is expected to reduce the number of administrations, prolong the administration interval, reduce the cost of medication, and improve patient compliance.

Claims (18)

An intraocular implant of axitinib, characterized in that the implant comprises axitinib and a biodegradable polymer. The intraocular implant according to claim 1, wherein the biodegradable polymer is selected from the group consisting of poly(lactic-co-glycolic acid) (PLGA) copolymer, polylactide (PLA), polyethylene One or more of lactide (PGA) and polyethylene glycol (PEG). The intraocular implant of claim 2, wherein the biodegradable polymer is PLGA. The intraocular implant of claim 3, wherein the biodegradable polymer is ester-terminated PLGA. The intraocular implant according to claim 4, wherein the molecular weight of the ester-terminated PLGA is 15,000-100,000, preferably 15,000-70,000, and most preferably 15,000 or 70,000. The intraocular implant of claim 5, wherein the ratio of glycolic acid and lactic acid monomers in the ester-terminated PLGA is 15:85 to 85:15, more preferably 50:50. The intraocular implant according to claim 6, characterized in that the intraocular implant comprises 20%-90% by weight of axitinib, preferably 30%-80% by weight of axitinib Axitinib, preferably 40%-70% axitinib by weight, preferably 40%-50% axitinib by weight or 50%-70% axitinib, preferably 45% by weight %-50% axitinib. The intraocular implant according to claim 7, characterized in that the intraocular implant comprises 30%-80% axitinib and 20%-70% ester-terminated PLGA, preferably 40% by weight. %-70% axitinib and 30%-50% ester-terminated PLGA, preferably, 40%-50% axitinib and 40%-50% ester-terminated PLGA or preferably 50%-70% axitinib Axitinib and 30%-50% ester-terminated PLGA, preferably comprising by weight 50% axitinib and 50% ester-terminated PLGA or 70% axitinib and 30% ester-terminated PLGA. The intraocular implant according to claim 8, characterized in that the weight of axitinib contained in the implant is 200 micrograms to 1.5 mg, preferably 200 micrograms to 1 mg or 1 mg to 1.5 mg, Preferably 200 micrograms, 400 micrograms, 500 micrograms, 600 micrograms, 700 micrograms, 1 mg or 1.5 mg. The intraocular implant according to claim 8, characterized in that the intraocular implant further comprises a release modifier comprising hydroxypropyl methylcellulose, hyaluronic acid and sodium salts thereof , poloxamer, polyethylene glycol, polyether, polyvinyl alcohol, polyvinylpyrrolidone, lactose, mannitol, glucose, maltose, sodium chloride, potassium chloride, magnesium carbonate, sodium bicarbonate, potassium bicarbonate or One or more of sucrose, preferably the release modifier includes one or more of polyethylene glycol, mannitol, sodium chloride, polyvinylpyrrolidone, preferably the release modifier includes mannitol, chlorine One or more of sodium chloride and polyvinylpyrrolidone. The intraocular implant according to claim 10, characterized in that the content of the release modifier is 0%-10% by weight, preferably 2.5%-10%, preferably 5%-10%, preferably 10% . The implant according to any one of claims 1-11, characterized in that the implant is for vitreous administration. A method for preparing the intraocular implant comprising any one of claims 1-12, characterized in that the preparation method comprises the steps of: physically mixing Axitinib with PLGA and/or a release modifier , add the above mixture into a twin-screw hot melt extruder, gradually heat up, and set the temperature between 90°C and 180°C, extrude, and cut the extrudate to the desired length after the extrusion is completed. The preparation method according to claim 13, wherein the temperature setting range is between 130°C and 180°C. The preparation method according to claim 13, characterized in that the preparation method comprises the steps of: physically mixing Axitinib with PLGA and/or a release modifier, adding the mixture into a twin-screw hot-melt extruder, Gradually heat up, the temperature setting range is between 90 ° C and 180 ° C, extrude, pulverize the rod shaped by the first extrusion, add it to the twin-screw extruder again, and perform the second extrusion. After pressing is complete, cut the extrudate to the desired length. The preparation method according to claim 13, characterized in that the preparation method comprises the steps of: pulverizing axitinib, physically mixing it with PLGA and/or a release modifier, and adding the mixture into a twin-screw hot-melt extruder In the process, the temperature is gradually increased, and the temperature is set in the range of 90°C to 180°C, and the extrusion is performed. After the extrusion is completed, the extrudate is cut to the desired length. The preparation method according to claim 13, characterized in that the preparation method comprises the steps of: pulverizing Axitinib and PLGA respectively, and/or physically mixing the release regulator, adding the above mixture into twin-screw hot melt extrusion In the machine, the temperature is gradually increased, and the temperature setting range is between 90 ° C and 180 ° C, and the extrusion is performed. After the extrusion is completed, the extrudate is cut to the desired length. Use of any one of the pharmaceutical compositions of claims 1-17 in the preparation of a medicament for eye diseases, preferably glaucoma, cataract, retinal vein occlusion, uveitis, diabetic macular edema and age associated macular degeneration.

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