Opthalmic composition of bevacizumab

Abstract

This invention relates to an ophthalmic composition for bevacizumab, which is safe and non-toxic in a single-use pre-filled syringe and is effective when administered within its expiration date. It is characterized by being an ophthalmic solution suitable for intravitreal use within two years of the product's expiration date, capable of controlling the number of non-visible particulate matter with significantly reduced endotoxin levels and low aggregation.

Description

[Technical Field] This invention relates to the fields of biotechnology and drug delivery. In particular, this invention relates to an ophthalmic composition for bevacizumab and an apparatus comprising the composition. [Previous Technology] Bevacizumab is a recombinant humanized monoclonal antibody containing a human antibody framework region and a mouse antibody complementarity-determining region. This mouse antibody binds to and inhibits the biological activity of human vascular endothelial growth factor (VEGF). Bevacizumab is licensed for the treatment of various cancers, including metastatic colorectal cancer (mCRC), non-squamous non-small cell lung cancer (NSCLC), metastatic breast cancer (MBC), metastatic renal cell carcinoma (mRCC), glioblastoma, and advanced cervical cancer. In these indications, bevacizumab has demonstrated safety and efficacy in patient survival. Furthermore, it is used as an off-label drug for the treatment of wet age-related macular degeneration (wAMD), choroidal neovascularization (CNV), diabetic macular edema (DME), retinal vein occlusion (RVO), and neovascular glaucoma (NVG). Alternatively, another monoclonal antibody, ranibizumab, is also on the market. It is approved by the U.S. Food and Drug Administration for the treatment of wet age-related macular degeneration, choroidal neovascularization, and diabetic macular edema. Although there appears to be no difference in efficacy and safety between bevacizumab and ranibizumab, multiple studies have demonstrated a significant cost advantage if bevacizumab is used instead of ranibizumab. Because bevacizumab is approved for cancer treatment, it is available in 100 mg or 400 mg vials designed to provide an individual with a full dose based on weight. When bevacizumab is used for wet age-related macular degeneration, ophthalmologists often administer relatively small doses repeatedly from a vial, as low as 1.25 mg per dose (multiple-dose administration). Any medication intended for intravitreal administration needs to be prepared and maintained in a sterile environment. There is a risk of contamination when the reconstituted solution is drawn from the device to prepare the injection in the vial before administration. Contaminated medication puts patients at risk of further infections, such as infectious endophthalmitis or alpha-hemolytic streptococcal infection. In some parts of the world, doctors draw the required medication from the original vial, and many consecutive patients use the same vial. This practice has proven unsafe for patients because the increasing number of punctures from the same vial increases the chance of contamination and infection. Furthermore, dosage may be inaccurate due to human error by healthcare professionals or caregivers. In some countries, dispensing pharmacies repackage multiple syringes from the original vials. The need to repackage medication from available vials into smaller doses increases the potential for contamination, especially since maintaining sterile conditions is difficult during such operations, and there is no way to verify whether dispensing pharmacies followed these procedures. The U.S. Food and Drug Administration (FDA) has documented and issued warnings regarding the use of sterility and dose control by these dispensing pharmacies in some cases, but this practice remains widespread. Furthermore, due to sterility issues and U.S. concerns, some dispensing pharmacies have recalled repackaged bevacizumab syringes. The FDA is now warning healthcare professionals not to use repackaged bevacizumab injections for intraocular use. In addition, bevacizumab is formulated for intravenous administration, so standard particle size distribution is crucial. Uncontrolled particles can cause irritation and / or inflammation in sensitive organs such as the eye. It has been reported that the particle size distribution of these repackaged injectable ophthalmic solutions exceeds the limits set by the United States Pharmacopeia (USP) and is often not standardized or uniform across different dispensing pharmacies. Besides safety concerns, the drug itself may have various effects related to the aliquoting, handling, and dispensing of the product. Deterioration may also depend on the specific packaging used, and the decreased stability of repackaged bevacizumab may be related to storage time. In addition to the potential safety risks associated with these changes, the drug's efficacy may also be affected. Similarly, the storage stability of repackaged bevacizumab is a major concern. The Drug Control Authority of India has issued guidelines on repackaged bevacizumab, recommending that repackaged bevacizumab syringes be used under specified conditions for no more than 14 days to prevent deterioration and denaturation. However, there is no effective mechanism to determine the duration of reconstruction and / or denaturation or deterioration during storage. Further reconstruction of the packaging product used for dosing is a complex and cumbersome process and may not always result in a homogeneous solution. Furthermore, there have been reports that counterfeit Avastin is being tracked internationally as global supply chains become increasingly complex, proving to be increasingly challenging. Implementing stringent regulatory guidelines can reduce some of the problems associated with bevacizumab being formulated into low doses for intravitreal use and counterfeit bevacizumab. However, existing technology issues such as limitations in bacterial endotoxin testing (BET) and particulate matter testing (PMT) require specific technical solutions and manufacturing controls to eliminate existing technology problems and provide products acceptable for intravitreal injection of ophthalmic drugs. Reports of adverse events associated with intravitreal administration of bevacizumab are increasing, including an increased incidence of aseptic ocular inflammation, infectious endophthalmitis, and elevated intraocular pressure. Some reported adverse events related to intravitreal administration of bevacizumab may be related to the manufacturing process of the bevacizumab or to a decrease in drug quality due to repackaging into plastic syringes. Furthermore, silica droplets generated by the needles and syringes used for intravitreal injection exacerbate the situation. The following are issues related to the product quality of the existing technology product: Bevacizumab currently only meets intravenous quality standards, and does not meet the more stringent ophthalmic standards regarding the levels of non-visible particulate matter and endotoxins. In the case of intravitreal injection, the presence of non-visible particulate matter and / or silicone oil is serious (accumulation in the eye, leading to potentially serious intraocular inflammation). From a microbiological and physicochemical perspective, there is no data to consider the quality of the repackaged product. Compatibility between the solution and the master packaging has also not been considered. The shelf life setting of the repackaged bevacizumab, and its impact on safety and quality in most parts of the world, has not been examined. Therefore, there is an urgent need for ophthalmic compositions containing bevacizumab that are safe, non-toxic, and effective for a single application with a long effective period. [Summary of the Invention] One object of the present invention is to provide an ophthalmic composition for bevacizumab having controllable particulate matter, bacterial endotoxins, and aggregates within its effective period, a method for preparing the composition, and a delivery device for the composition, the device being safe, non-toxic, and effective for single-dose administration. This invention discloses an ophthalmic composition of bevacizumab, administered in the form of a single-use pre-filled syringe. The ophthalmic composition of this invention controls bacterial endotoxin, particulate matter, and aggregate limits over its 2-year shelf life. This invention also provides a method for obtaining the active ingredient, bevacizumab, in which the amounts of bacterial endotoxin and particulate matter are negligible or zero, and the composition of this invention, when formulated, has very low aggregate, bacterial endotoxin, and particulate matter content over its shelf life, and is suitable for intravitreal use. This invention also provides an apparatus, such as a pre-filled syringe or cartridge, containing the ophthalmic composition of bevacizumab.

Implementation Method

Claims

1. An eye drop composition comprising: Bevacizumab in the range of 24 mg / mL to 26 mg / mL; A buffer in the range of 40 mM to 60 mM; A stabilizer in the range of 40 mg / mL to 70 mg / mL; and a nonionic surfactant in the range of 0.2 mg / mL to 0.6 mg / mL; a. wherein the bacterial endotoxin test is in the range of 0.001 to 0.16 EU / mg; b. The particulate matter is: (i) 1 to 50 particles with a diameter ≥ 10 μm per ml; (ii) 0 to 5 particles with a diameter ≥ 25 μm per ml; and (iii) 0 to 2 particles with a diameter ≥ 50 μm per ml; c. The aggregate size of the ophthalmic composition within 2 to 3 years ranges from 0.1% to 5%; d. The buffer is selected from the group consisting of phosphate, citrate, acetate, histidine, succinate, gluconate, and glycine, and the pH of the buffer is in the range of 6.0 to 7.0; e. The stabilizer is selected from monosaccharides, disaccharides, trisaccharides, polysaccharides, sugar alcohols, reducing sugars, and non-reducing sugars; f. The nonionic surfactant is polysorbate 20 or polysorbate 80; g. The ophthalmic composition is administered in the form of a single-use pre-filled syringe.

2. A method for preparing an ophthalmic composition as described in claim 1, comprising the following steps: i. Chinese hamster ovary cells used to produce bevacizumab via continuous fermentation; ii. The cell-free harvest obtained from Chinese hamster ovary cell culture was subjected to adsorption-based depth filtration for further clarification; iii. The sample from step ii was concentrated using single-pass tangential flow filtration to obtain a concentrated harvest; iv. The concentrated harvest from step iii is subjected to protein A chromatography to capture bevacizumab and obtain an eluent containing partially purified bevacizumab; v. Deactivate the virus from the eluent of step iv at low pH to obtain a virus-deactivated sample; vi. Further purify the virus-deactivated sample from step v by cation exchange chromatography to obtain an eluent primarily containing highly purified bevacizumab; vii. Perform anion exchange chromatography on the eluent of step vi to remove trace impurities, including host cell proteins, host cell DNA, and endotoxins; viii. Perform virus reduction filtration on the eluent of step vii to completely remove the virus; ix. Concentrate and dialyze the sample from step viii to obtain bevacizumab; And x. Formulate the sample obtained from step ix to obtain the ophthalmic composition of the present invention as claimed in claim 1, wherein the ophthalmic composition comprises: a bevacizumab in the range of 24 mg / mL to 26 mg / mL; a buffer in the range of 40 mM to 60 mM; a stabilizer in the range of 40 mg / mL to 70 mg / mL; and a nonionic surfactant in the range of 0.2 mg / mL to 0.6 mg / mL; a. wherein the bacterial endotoxin test is in the range of 0.001 to 0.16 EU / mg; b. wherein the particulate matter is: (i) 1 to 50 particles per ml with a diameter ≥10 μm; (ii) 0 to 5 particles per ml with a diameter ≥25 μm; and (iii) 0 to 2 particles per ml with a diameter ≥50 μm; c. wherein the aggregates of the ophthalmic composition in the range of 0.1% to 3.5% over 2 years; d. The buffer is selected from the group consisting of phosphate, citrate, acetate, histidine, succinate, gluconate, and glycine, and the pH of the buffer is in the range of 6.0 to 7.0; e. The stabilizer is selected from monosaccharides, disaccharides, trisaccharides, polysaccharides, sugar alcohols, reducing sugars, and non-reducing sugars; f. The nonionic surfactant is polysorbate 20 or polysorbate 80.

3. The preparation method as described in claim 2, wherein the low viral deactivation is carried out in the pH range of 3.0 to 5.

0.

4. The preparation method as described in claim 2, wherein the stationary phase for cation exchange chromatography is selected from the group consisting of SO3-sulfonyl isobutyl, SO3-sulfonyl ethyl, sulfonyl propyl, and carboxymethyl, and the stationary phase for anion exchange chromatography is selected from the group consisting of diethylaminoethyl, quaternary ammonium, polyquaternary ammonium salt, and N-phenylmethyl-N-methylethanolamine, and dialysis is performed in the presence of TFF-II dialysis buffer.

5. The ophthalmic composition as claimed in claim 1, wherein the concentration of bevacizumab in the ophthalmic composition is in the range of 25 mg / ml to 26 mg / ml.

6. The ophthalmic composition as claimed in claim 1, wherein the concentration of the buffer is in the range of 50 mM to 60 mM.

7. The ophthalmic composition as claimed in claim 1, wherein the stabilizer is a sugar selected from glucose, sucrose, trehalose, lactose, fructose, maltose, polydextrose, glycerol, polydextrose, erythritol, glycerol, arabinitol, xylitol, sorbitol, mannitol, melibiose, pinetriose, raffinose, mannitol, stachyose, maltose, lactulose, maltulose, glucol, maltitol, lactitol, isomaltose, and in the range of 45 mg / mL to 65 mg / mL.

8. The ophthalmic composition as claimed in claim 1, wherein the nonionic surfactant is in the range of 0.3 mg / mL to 0.5 mg / mL.

9. The ophthalmic composition as claimed in claim 1, wherein it is administered in a pre-filled syringe at a predetermined dose of 140 mL to 200 mL and a dose of 50 mL.

10. Use of the ophthalmic composition as claimed in claim 1 in the preparation of a medicament for treating ocular diseases, selected from age-related wet macular degeneration, choroidal neovascularization, retinal angioma hyperplasia, pathological myopia, vascular scars, Best's disease, adult yolk sac dystrophy, central serous chorioretinopathy, punctate intrachorioretinopathy, multifocal choroiditis, suspected ocular tissue lasmolysis, choroidal osteoma, toxoplasmosis, uveitis, pseudotumor of the brain, spontaneous perimasculinic retinal neovascularization, proliferative diabetic retinopathy, sickle cell retinopathy, retinopathy of prematurity, Eales' disease, macular edema, diabetic retinopathy, central retinal vein occlusion, branch retinal vein occlusion, pseudolens, uveitis, thromboangiitis obliterans, and retinal diseases. This group comprises: membranous pigmentary degeneration, neovascular glaucoma, central retinal vein occlusion, branching retinal vein occlusion, proliferative diabetic retinopathy, central retinal artery occlusion, ocular ischemic syndrome, radiation-induced, radiation-induced optic neuropathy, radiation-induced retinopathy, choroidal metastasis of breast cancer, melanoma-associated neovascularization, aortic aneurysm, vascular proliferative tumor, Koplik's disease, adjacent papillary capillary hemangioma, spontaneous macular telangiectasia, multiple polypoid choroidal angiopathy, central serous chorioretinopathy, non-arterial anemic optic neuropathy, herpetic keratotic neovascularization, cicatricial pemphigoid keratotic neovascularization, posterior capsule neovascularization, corneal transplant rejection neovascularization, dry eye-related corneal neovascularization, bleb repair, and adjunctive treatments for glaucoma filtration surgery.

11. A kit comprising an ophthalmic composition as described in any one of claims 1 to 8, the kit further comprising a pre-filled syringe and a needle in the size range of 29 to 34.

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