US20200345636A1 - Artificial Vitreous Humor for the Investigation of Drugs and Drug Formulations - Google Patents

Artificial Vitreous Humor for the Investigation of Drugs and Drug Formulations Download PDF

Info

Publication number
US20200345636A1
US20200345636A1 US16/855,893 US202016855893A US2020345636A1 US 20200345636 A1 US20200345636 A1 US 20200345636A1 US 202016855893 A US202016855893 A US 202016855893A US 2020345636 A1 US2020345636 A1 US 2020345636A1
Authority
US
United States
Prior art keywords
vitreous humor
composition
artificial
artificial vitreous
substance
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Abandoned
Application number
US16/855,893
Other languages
English (en)
Inventor
Ulrike Altenburger
Sulabh Pravinchandra PATEL
Gregoire Schwach
Pankaj SHENDE
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Hoffmann La Roche Inc
Original Assignee
Hoffmann La Roche Inc
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Hoffmann La Roche Inc filed Critical Hoffmann La Roche Inc
Assigned to F. HOFFMANN-LA ROCHE AG reassignment F. HOFFMANN-LA ROCHE AG ASSIGNMENT OF ASSIGNORS INTEREST (SEE DOCUMENT FOR DETAILS). Assignors: ALTENBURGER, Ulrike, PATEL, Sulabh Pravinchandra, SCHWACH, GREGOIRE
Assigned to F. HOFFMANN-LA ROCHE AG reassignment F. HOFFMANN-LA ROCHE AG ASSIGNMENT OF ASSIGNORS INTEREST (SEE DOCUMENT FOR DETAILS). Assignors: SHENDE, Pankaj
Assigned to HOFFMANN-LA ROCHE INC. reassignment HOFFMANN-LA ROCHE INC. ASSIGNMENT OF ASSIGNORS INTEREST (SEE DOCUMENT FOR DETAILS). Assignors: F. HOFFMANN-LA ROCHE AG
Publication of US20200345636A1 publication Critical patent/US20200345636A1/en
Priority to US18/298,991 priority Critical patent/US20240074972A1/en
Abandoned legal-status Critical Current

Links

Images

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/0012—Galenical forms characterised by the site of application
    • A61K9/0019—Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/02—Inorganic compounds
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
    • A61K47/16—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing nitrogen, e.g. nitro-, nitroso-, azo-compounds, nitriles, cyanates
    • A61K47/18—Amines; Amides; Ureas; Quaternary ammonium compounds; Amino acids; Oligopeptides having up to five amino acids
    • A61K47/183—Amino acids, e.g. glycine, EDTA or aspartame
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
    • A61K47/26—Carbohydrates, e.g. sugar alcohols, amino sugars, nucleic acids, mono-, di- or oligo-saccharides; Derivatives thereof, e.g. polysorbates, sorbitan fatty acid esters or glycyrrhizin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
    • A61K47/36—Polysaccharides; Derivatives thereof, e.g. gums, starch, alginate, dextrin, hyaluronic acid, chitosan, inulin, agar or pectin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
    • A61K47/42—Proteins; Polypeptides; Degradation products thereof; Derivatives thereof, e.g. albumin, gelatin or zein
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/0012—Galenical forms characterised by the site of application
    • A61K9/0048—Eye, e.g. artificial tears
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/08—Solutions

Definitions

  • the invention relates to artificial vitreous humor compositions, a method for the production of an artificial humor, a method for analyzing the behavior of a substance and a kit of parts.
  • intravitreal injections of drugs have proven successful.
  • a drug formulation is directly injected into the vitreous humor (VH), that is, the clear gel filling the space between lens and retina of the eyeball of for example humans.
  • VH vitreous humor
  • the fraction of an administered dose of an unchanged drug that reaches the retina is high, thus intravitreal injections usually have high bioavailability.
  • VH when not in its natural environment, degenerates fast and the pH value of VH may increase rapidly due to accumulation of degenerated products in the VH. Thus, tests may not represent the actual situation in an eye of a living subject, especially not over long periods of time, e.g. several days.
  • the physiological pH value of VH may be stabilized by applying a buffering system. Therein, degradation products are allowed to leave the VH through a semi-permeable membrane into a buffer solution.
  • VH intravitreal
  • DDS drug-loaded long acting delivery
  • the therapeutic agent remains in the VH and gets exposed at physiological pH and temperature for significantly longer duration. Therefore, it is of prime interest to study the stability of these agents in VH at physiological conditions. Physical and chemical variability of the natural human VH is accounted from disease condition and age, which may have direct impact on the stability predictions.
  • VH of animal origin e.g. rabbit, pig, monkey or others
  • variabilities e.g. relating to variabilities in viscosity and/or density.
  • VH is composed of various small and large inorganic and organic molecules which can be largely categorized as buffer system, small molecular weight components (e.g. salts and ions) and matrix forming polymers (e.g. collagen and hyaluronic acid).
  • small molecular weight components e.g. salts and ions
  • matrix forming polymers e.g. collagen and hyaluronic acid
  • the remaining constituents 1-2%) are mainly collagen, hyaluronic acid, proteins and small mollecular weight components such as ascorbic acid, glutathione, glucose, lactate, xanthine, hypoxanthine, creatinine and urea as disclosed in Kleinberg et al. (Vitreous substitutes: a comprehensive review, Sury Ophthalmol. 2011 July-August; 56(4): 300-23) and Amith Mulla (Role of vitreous humor biochemistry in forensic pathology, University of Saskatchewan, Thesis, 2005).
  • novel compositions of aVH comprising a buffer system, small molecular weight (SMW) components and large molecular weight (LMW) components such as matrix forming polymers, which can then serve as an in-vitro model for the prediction of drug or drug formulation stability, and drug release behavior from DDS systems following IVT delivery.
  • SSW small molecular weight
  • LMW large molecular weight
  • the novel aVH would prove as the simplest yet very efficient real-time in-vitro experimental medium for stability and release prediction.
  • the novel aVH compositions as described herein may serve as research tool but also as quality control tool.
  • an artificial vitreous humor composition comprising a phosphate buffer.
  • the phosphate buffer has a pH value in the range from 7.0 to 7.7, particularly from 7.1 to 7.6, more particularly from 7.2 to 7.5. Most particularly the phosphate buffer has a pH value of 7.4.
  • the phosphate buffer may be selected from the group consisting of phosphate buffer saline (PBS), phosphate buffer saline including Tween (PBST), phosphate buffer saline including Tween and sodium azide (PBSTN) and phosphate buffer saline including sodium azide (PBSN).
  • the phosphate buffer may be phosphate buffer saline (PBS).
  • PBS phosphate buffer saline
  • the phosphate buffer saline may be in the range particularly from 0.001 to 0.2 M, more particularly from 0.003 to 0.05, most particularly from 0.005 to 0.02 M.
  • the phosphate buffer saline preferably has a pH value of 7.4.
  • a preferred buffer according to the invention is 0.01 M PBS, pH 7.4 comprising 8 gm/L NaCl, 0.2 gm/L KCl, 1.44 gm/L Na 2 HPO 4 and 0.24 gm/L KH 2 PO 4 .
  • the artificial vitreous humor composition may comprise at least one small molecular weight component.
  • the small molecular weight component may be selected from the group consisting of creatinine, glucose, urea, xanthine, hypoxanthine, sodium lactate and glutathione.
  • the artificial vitreous humor composition may comprise at least one of up to 578 ⁇ M creatinine, up to 28.1 mM glucose, up to 58.5 mM urea, 200 to 1630 ⁇ M xanthine, 100 to 800 ⁇ M hypoxanthine, 0.1 to 23 mM sodium lactate, 50 to 300 ⁇ M glutathione.
  • the artificial vitreous humor composition comprises 64.6 ⁇ M creatinine, 2.2 mM glucose, 7.6 mM urea, 580 ⁇ M xanthine, 309 ⁇ M hypoxanthine, 10.5 mM sodium lactate and 200 ⁇ M glutathione.
  • the artificial vitreous humor composition may comprise 20 to 300 mg/L type II collagen and/or 0.03 to 0.9% w/v sodium hyaluronate.
  • Collagen and/or sodium hyaluronate serve as matrix forming components in the artificial vitreous humor.
  • the artificial vitreous humor compositions comprises 40 mg/L type II collagen and/or 0.6% w/v sodium hyaluronate.
  • the artificial vitreous humor composition may not comprise ascorbic acid.
  • a further aspect of the invention relates to a method for the production of an artificial vitreous humor composition, preferably as described herein.
  • the method comprises step (i), wherein stock solutions of glucose, creatinine, sodium lactate, glutathione and urea in water, a stock solution of hypoxanthine in formic acid:water of 2:1 ratio, and a stock solution of xanthine sodium in 1M NaOH are provided.
  • the method further comprises step (ii) of mixing of the stock solutions of step (i) in a phosphate buffer with a pH value in the range from 7.0 to 7.7, particularly from 7.1 to 7.6, more particularly from 7.2 to 7.5, most particularly a pH value of 7.4, and pH adjustment to the pH value of the phosphate buffer.
  • step (iii) hyaluronic acid is added to the mixture obtained in step (iii) and the mixture is stirred at 2 to 8° C. for at least 4 hours. Particularly, the mixture is stirred until the hyaluronic acid is completely dissolved.
  • the method further comprises a step (iv), wherein collagen type-II is added, preferably with constant stirring, to the mixture obtained in step (iii).
  • the mixture obtained in step (iv) is optionally sterile filtered.
  • the phosphate buffer may be selected from the group consisting of phosphate buffer saline (PBS), phosphate buffer saline including Tween (PBST), phosphate buffer saline including Tween and sodium azide (PBSTN) and phosphate buffer saline including sodium azide (PBSN).
  • PBS phosphate buffer saline
  • the phosphate buffer saline may be in the range particularly from 0.001 to 0.2 M, more particularly from 0.003 to 0.05, most particularly from 0.005 to 0.02 M.
  • the phosphate buffer saline preferably has a pH value of 7.4.
  • a preferred buffer according to the invention is 0.01 M PBS, pH 7.4 comprising 8 gm/L NaCl, 0.2 gm/L KCl, 1.44 gm/L Na 2 HPO 4 and 0.24 gm/L KH 2 PO 4 .
  • the artificial vitreous humor composition may comprise at least one small molecular weight component.
  • the small molecular weight component may be selected from the group consisting of creatinine, glucose, urea, xanthine, hypoxanthine, sodium lactate and glutathione.
  • the aVH composition comprises 64.6 ⁇ M creatinine, 2.2 mM glucose, 7.6 mM urea, 580 ⁇ M xanthine, 309 ⁇ M hypoxanthine, 10.5 mM sodium lactate and 200 ⁇ M glutathione.
  • the artificial vitreous humor composition may comprise 20 to 300 mg/L type II collagen and/or 0.03 to 0.9% w/v sodium hyaluronate.
  • Collagen and/or sodium hyaluronate serve as matrix forming components in the artificial vitreous humor.
  • the artificial vitreous humor compositions comprises 40 mg/L type II collagen and/or 0.6% w/v sodium hyaluronate.
  • the artificial vitreous humor compositions comprises 40 mg/L type II collagen and 0.6% w/v sodium hyaluronate.
  • the artificial vitreous humor composition may not comprise ascorbic acid.
  • a further aspect of the invention relates to a method for analyzing the behavior of a substance applied to an artificial vitreous humor composition, preferably as described herein.
  • the method comprises step (i), wherein an artificial vitreous humor composition, preferably as described herein, is provided in an in-vitro environment.
  • An in-vitro environment according to the invention may be a vial, preferably a glass vial or plastic vial, well, container, depreshion dish, ex-vivo intravitreal (ExVit) model as described in Patel et al., Evaluation of protein drug stability with vitreous humor in a novel ex-vivo intraocular model, European Journal of Pharmaceutics and Biopharmaceutics.
  • the method further comprises step (ii), wherein the substance to be analyzed is applied to the artificial vitreous humor.
  • the substance may be administered to the artificial vitreous humor composition.
  • the substance may be injected to the artificial vitreous humor composition or diffused into the artificial vitreous humor composition or released from a long-acting sustained release delivery (DDS) system into the artificial vitreous humor composition.
  • DDS sustained release delivery
  • the method comprises step (iii) wherein at least one property of the applied substance is determined.
  • the property of the substance to be applied is known prior to the application to the vitreous humor, a comparison of the property of the substance before and after application to the artificial vitreous humor is feasible; e.g. in order to analyze the stability of the applied substance when present in the aVH, bioavailability, release from DDS system and degradation of the DDS system (Blanco et al., Protein encapsulation and release from poly(lactide-co-glycolide) microspheres: effect of the protein and polymer properties and of the co-encapsulation of surfactants, European journal of pharmaceutics and biopharmaceutics. 1998 May; 45(3): 285-94) within the aVH.
  • the property of the substance may be determined immediately and/or in intervals in order to follow the change of the property over time.
  • the substance to be applied may be at least one of a macromolecule, a protein, e.g. an antibody, a peptide, an oligonucleotide, an aptamer, a drug formulation, an excipient, a small molecule, a drug delivery system (biodegradable and nonbiodegradable), or a combination thereof.
  • a macromolecule e.g. an antibody, a peptide, an oligonucleotide, an aptamer, a drug formulation, an excipient, a small molecule, a drug delivery system (biodegradable and nonbiodegradable), or a combination thereof.
  • a protein e.g. an antibody, a peptide, an oligonucleotide, an aptamer, a drug formulation, an excipient, a small molecule, a drug delivery system (biodegradable and nonbiodegradable), or a combination thereof.
  • biodegradable and nonbiodegradable biodegradable
  • the substance to be applied may comprise molecules having a size in a range between 100 Da and 1800 kDa, particularly in a range between 1 kDa and 500 kDa, more particularly between 4 kDa and 200 kDa, most particularly between 10 kDa and 175 kDa.
  • the at least one property of the applied substance is selected from the group consisting of stability, bioavailability, release from DDS system and degradation of DDS system.
  • the applied substance may be left in the artificial vitreous humor composition for up to 360 days, particularly up to 180 days, more particularly up to 90 days, prior to step (iii).
  • the property of the substance may be determined in intervals in order to follow the change of the property with time.
  • step (iii) may define the last determination in time of the at least one property.
  • the artificial vitreous humor may be maintained at a constant temperature prior to step (iii), particularly at a temperature in the range from 32 to 38° C., more particularly from 35 to 37° C., most particularly from 36 to 37° C.
  • a further aspect of the invention relates to a method for analyzing the behavior of an artificial humor composition as described herein, wherein a substance is applied to the artificial humor composition.
  • the method comprises step (i), wherein an artificial vitreous humor composition, preferably as described herein, is provided in an in-vitro environment.
  • An in-vitro environment according to the invention may be a vial, preferably a glass vial or plastic vial, well, container, depreshion dish, ex-vivo intravitreal (ExVit) model as described in Patel et al., Evaluation of protein drug stability with vitreous humor in a novel ex-vivo intraocular model, European Journal of Pharmaceutics and Biopharmaceutics.
  • the method further comprises step (ii), wherein a substance is applied to the artificial vitreous humor.
  • the substance may be administered to the artificial vitreous humor composition.
  • the substance may be injected to the artificial vitreous humor composition or diffused into the artificial vitreous humor composition or released from a long-acting sustained release delivery (DDS) system into the artificial vitreous humor composition.
  • the method comprises step (iii) wherein at least one property of the artificial humor composition is determined.
  • the property of the artificial humor composition is known prior to the application of the substance, a comparison of the property of the the artificial humor composition before and after application of the substance is feasible; e.g.
  • the method allows an assessment of the tolerability of the artificial humor composition in view of the applied substance, particularly by determining the change of at least one of the group consisting of viscosity, turbidity, osmolality.
  • the property of the the artificial humor composition may be determined immediately and/or in intervals in order to follow the change of the property over time.
  • the substance to be applied may be at least one of a macromolecule, a protein, e.g.
  • the substance to be applied may comprise molecules having a size in a range between 100 Da and 1800 kDa, particularly in a range between 1 kDa and 500 kDa, more particularly between 4 kDa and 200 kDa, most particularly between 10 kDa and 175 kDa.
  • the applied substance may be left in the artificial vitreous humor composition for up to 360 days, particularly up to 180 days, more particularly up to 90 days, prior to step (iii).
  • the property of the the artificial humor composition may be determined in intervals in order to follow the change of the property with time.
  • step (iii) may define the last determination in time of the at least one property.
  • the artificial vitreous humor may be maintained at a constant temperature prior to step (iii), particularly at a temperature in the range from 32 to 38° C., more particularly from 35 to 37° C., most particularly from 36 to 37° C.
  • a further aspect of the invention relates to a method for analyzing the behavior of a substance in a artificial humor as described herein and/or the behavior of an artificial humor composition as described herein.
  • a further aspect of the invention relates to an artificial vitreous humor composition as described herein produced by the method of production as described herein.
  • a further aspect of the invention relates to a kit of parts for the production of an artificial vitreous humor composition, particularly as described herein.
  • the kit of parts comprises several components.
  • Component (A) is a phosphate buffer, particularly phosphate buffer saline (PBS), more particularly 0.01 M phosphate buffer saline (PBS), having a pH value in the range from 7.0 to 7.7, particularly from 7.1 to 7.6, more particularly from 7.2 to 7.5. most particularly a pH value of 7.4.
  • Component (B) comprises at least one selected from the group consisting of creatinine, glucose, urea, xanthine, hypoxanthine, sodium lactate, glutathione.
  • component (B) comprises a mixture of creatinine, glucose, urea, xanthine, hypoxanthine, sodium lactate, glutathione or component (B) is a selection of components referred to as (B1) creatinine, (B2) glucose, (B3) urea, (B4) xanthine, (B5) hypoxanthine, (B6) sodium lactate and (B7) glutathione.
  • Component (C) comprises type II collagen and/or sodium hyaluronate.
  • component (C) is a selection of components referred to as (C1) type II collagen and (C2) sodium hyaluronate.
  • Optional component (D) is instructions for use.
  • a further aspect of the invention relates to the use of an artificial vitreous humor composition as described herein in a method for analyzing the behavior of a substance in the artificial humor composition as described herein.
  • a further aspect of the invention relates to the use of an artificial vitreous humor composition as described herein in a method for analyzing the behavior of the artificial vitreous humor composition, when a substance is applied to the artificial vitreous humor composition.
  • a further aspect of the invention relates to the use of an artificial humor composition as described herein in a method for analyzing the behavior of a substance in the artificial composition as described herein and/or in a method for analyzing the behavior of the artificial humor composition, wherein a substance is applied to the artificial humor composition, as described herein.
  • a further aspect of the invention relates to the use of a phosphate buffer saline having a pH value in the range from 7.0 to 7.7, particularly from 7.1 to 7.6, more particularly from 7.2 to 7.5, most particularly a pH value of 7.4 for the production of an artificial vitreous humor composition as described herein.
  • FIG. 1 Absorbance analysis correlating to the amount of reduced ascorbic acid in three different simulated buffer media (SBM) prepared at different buffer strength after 1 week of incubation at 37° C.
  • SBM simulated buffer media
  • FIG. 2 Determination of the viscosity of isolated porcine vitreous humor.
  • FIGS. 3 a and 3 b Pictorial representations of the ExVit model utilized to compare protein stability in the natural VH vs artificial VH ( FIG. 3 a ) and of vial based model used to investigate protein stability in the artificial VH ( FIG. 3 b ).
  • FIGS. 4 a and 4 b Analysis of pH ( FIG. 4 a ) and osmolality ( FIG. 4 b ) of the PBS and vitreous humor (aVH and pVH) in ExVit or vial model with or without a monoclonal antibody (mAb) after incubation at 37° C. (Experiments were performed in triplicates and results are reported in mean ⁇ std. dev).
  • FIG. 5 Analysis of turbidity of the PBS and vitreous humor (aVH and pVH) in ExVit or vial model with or without mAb after incubation at 37° C. (Experiments were performed in triplicates and results are reported in mean ⁇ std. dev).
  • FIGS. 6 a and 6 b Physical stability of mAb in aVH (ExVit or vial model), pVH and PBS were investigated by SEC.
  • FIG. 6 a represents the main peak.
  • FIG. 6 b represents the higher molecular weight species (HMWS, %) (Experiments were performed in triplicates and results are reported in mean ⁇ std. dev).
  • FIGS. 7 a and 7 b Physical stability of mAb in aVH (ExVit or vial model), pVH and PBS was investigated by CE-SDS-NGS.
  • FIG. 7 a represents the main peak.
  • FIG. 7 b represents the deglycosylated variants (area %).
  • FIGS. 8 a and 8 b Binding affinity of mAb.
  • FIG. 8 a represents binding affinity to antigen-1.
  • FIG. 8 b represents the binding affinity to antigen-2 retrieved from stability samples following incubation in aVH (ExVit or vial model) and pVH.
  • FIG. 9 Cumulative Release (%) of Small Molecule (SM) active pharmaceutical ingredients (API) from various PLGA based LAD intravitreal implants.
  • SM Small Molecule
  • API active pharmaceutical ingredients
  • “Vitreous humor” as used in this application may be from natural or artificial sources. Natural vitreous humor may be from various animal species, for example porcine, bovine, canine, feline, from rabbits and non-human primates or may be human. Porcine vitreous humor is abbreviated pVH herein. Natural vitreous humor may for example be gained from eyes that have previously been removed.
  • Artificial vitreous humor preferably mimics human vitreous humor.
  • Artificial vitreous humor may be prepared from various polymeric materials, also called large molecular weight (LMW) components or matrix forming components, for example natural polymers such as but not limited to hyaluronic acid, alginate, agar, chitosan, gelatin, xanthan gum, pectins, collagen, or for example synthetic polymers such as but not limited to pluronic gel, polyvinyl alcohol, polyphosphazenes, any dimeric, trimeric or multimeric gelling polymers composed of PEG, PCL, PLA, PGA, PLGA, poly acrylamide, polyacrylic acid, as well as combinations of these polymers at different concentrations.
  • LMW large molecular weight
  • LMW components particularly the hyaluronic acid
  • Artificial vitreous humor may further comprise small molecular weight (SMW) components.
  • SMW small molecular weight
  • These small molecular weight components may be selected from the group comprising nitrogenous organic acids, sugars, reducing agents, such as urea, purine bases or derivatives thereof, salts of lactic acid and antioxidants, such as glutathione.
  • Vitreous humor may also be a mixture of artificial and natural vitreous humor or their components in different combinations.
  • a “buffer solution” is a solution preferably having or being able to maintain the pH value of the system, for example between pH 5.5 and pH 8.5, preferably between about pH 7.0 and about pH 7.6, more preferably at pH 7.4.
  • “Physiologically relevant buffer solution” is a solution preferably having or being able to maintain a pH value of the system between about pH 7.0 and about pH 7.6, more preferably at pH 7.2-7.4.
  • a buffer solution comprises salts.
  • the buffer solution may for example be a phosphate buffered saline (PBS), a bicarbonate buffer, Ringer's bicarbonate buffer, Ringer's lactate buffer, simulated body fluids, other isotonic solutions, cell culture medias, and any other physiologically representative buffers.
  • PBS phosphate buffered saline
  • PBS according to the invention may comprise 8 gm/L NaCl, 0.2 gm/L KCl, 1.44 gm/L Na 2 HPO 4 and 0.24 gm/L KH 2 PO 4 .
  • PBS preferably has a pH value of 7.4.
  • the term “behavior of a substance” relates to the change of a property of a substance when applied to the artificial vitreous humor or else to the artificial humor composition according to the invention.
  • the change of property of a substance can be a) chemical or physical changes, b) change in its binding to a target, c) transformation from one polymorphic form to another polymorphic form, d) change of physical state, e.g. liquid to gel.
  • a substance can be a therapeutic agent or two or more therapeutic agents, excipient, formulation (simple or sustained release) or placebo (simple or placebo of sustained release formulation).
  • the composition of artificial VH (aVH) allows simulating in vivo physiological conditions of natural VH.
  • the formulation's stability and release from DDS may be simulated, for human but also for animal eyes.
  • different diffusion and precipitation behavior may be tested for, for example, different proteins in different eye conditions (corresponding to more or less degenerated eyes, by age or by disease) for posterior eye tissue.
  • These conditions can be simulated and tested with the aVH and the method for analyzing the behavior of a substance according to the invention by varying the properties, e.g. composition and/or density and/or viscosity, of the aVH.
  • artificial vitreous humor and the method according to the invention allow to more realistically simulate the geometry of a natural environment, preferably of an eye environment.
  • the term “behavior of an artificial humor composition” relates to the change of a property of the artificial humor composition, when a substance is applied to the artificial vitreous humor according to the invention.
  • the change of property of the artificial humor composition can particularly be chemical or physical changes, and/or change of physical state, e.g. gel to liquid.
  • Porcine eyes were acquired from a local slaughter-house located near Zurich, Switzerland. Custom made side-by-side diffusion chambers were purchased from SES GmbH-Analytical Systems (Bechenheim, Germany) A diffusion controlling membrane, with molecular weight cut-off (MWCO) of 50 kDa (cat #131384), was procured from Spectrum lab (California, USA). Collagen (type-II) (cat #804001-sol) and sodium hyaluronate (cat #HA15M) were purchased from mdbioproducts and Lifecore Biomedical, respectively.
  • MWCO molecular weight cut-off
  • L-ascorbic acid (cat #A5960), xanthine sodium salt (cat #X3627), hypoxanthine (cat #H9636), creatinine (cat #C4255), urea (cat #51457), glucose (cat #G8270), L-glutathione (cat #G4251) and sodium lactate (cat #71718) were procured from Sigma Aldrich, USA.
  • Bispecific monoclonal antibody (mAb) was manufactured in a CHO cell line and provided by F. Hoffmann-La Roche AG, Basel. The mAb is a full length humanized mAb based on the IgG-1 format with an approximate molecular weight of 150 kDa.
  • mAb was formulated in histidine buffer (20 mM, pH 6.0) containing sugar and surfactant to provide tonicity and stability, respectively.
  • the small molecule with a MW ⁇ 600 Dalton and LogP of 3.24 was provided by F. Hoffmann-La Roche AG, Basel.
  • PLGA polymers were procured from Evonik, Parsippany, USA (RG756S PL(DL)GA 75:25 and RG753H PL(DL)GA 75:25).
  • phosphate buffer saline (0.01 M PBS, pH 7.4) was used in the buffer-compartment. All other reagents utilized in these studies were of analytical grade.
  • Natural VH is a liquid tissue mainly composed of water with small amounts of organic and inorganic components including matrix forming polymers.
  • the composition of VH is reported in literature.
  • aVH artificial VH
  • SBM simulated buffer medium
  • SBM small molecular weight
  • SBMs were sterile filtered with 0.22 ⁇ filter under laminar air flow. SBMs were then investigated for storage stability by incubating at 37° C. for 7 days. Buffer compatibility was evaluated by measuring pH and clarity. For pH estimation, samples (100 ⁇ L) were aseptically transferred in eppendorf tubes and analyzed by calibrated pH meter (827 pH Lab, Metrohm, Switzerland). Clarity of the SBMs was observed visually and the samples were classified as clear or turbid.
  • PBS based SBM compositions 10-17 remained clear after 7 days of incubation at 37° C. (Table 4). However, all of these compositions also showed acidic pH shift after 7 days.
  • SBM with or without sodium lactate, hypoxanthine and ascorbic acid, and SBM with higher buffer strength were investigated.
  • the SBMs were prepared as reported in Table 5.
  • SBMs were sterile filtered and incubated at 37° C. for one week and analyzed for pH and clarity. Samples were collected and analysis was performed as described above. Also, the amount of reduced ascorbic acid was analyzed in the samples using 1,1-diphenyl-2-picrylhydrazyl ( ⁇ , ⁇ -diphenyl- ⁇ -picrylhydrazyl (DPPH) assay. Briefly, 100 ⁇ L of ascorbic acid standard or test samples were incubated in presence of 1900 ⁇ L of DPPH solution at 37° C. for 30 min in dark. Samples were then analyzed for change in optical density (OD) at 517 nm against methanol as blank. The presence of ascorbic acid turns color of solution from violet to brownish-pale yellow.
  • OD optical density
  • compositions 18-20 were not sufficient to maintain the pH.
  • absence of sodium lactate and hypoxanthine or both also could not prevent the acidic pH shift (compositions 21-27).
  • SBM compositions 28-30, without ascorbic acid exhibited stable pH after 7 days.
  • SBMs with or without Ascorbic acid exhibited similar levels of absorbance at 517 nm suggesting absence of reduced form of ascorbic acid after 7 days of incubation at 37° C. Ascorbic acid might be oxidized in less than 7 days resulting in the pH shift to the acidic side. Hence, ascorbic acid was not included in the final SBM composition.
  • composition 28 of SBMs was found stable for 7 days without change in pH or clarity, it was further characterized for long-term storage stability at 37° C. Briefly, composition 28 was prepared and aseptically filtered under LAF. The sterile SBM was then transferred aseptically in 6 mL sterile glass vials. The glass vials were sealed and incubated at 37° C. for 3 months. The samples were collected at various time points such as, initial, week-1, week-2, week-5, week-8 and week-12, and analyzed for pH, clarity, density, viscosity and osmolality. For pH estimation, samples (100 ⁇ L) were aseptically transferred in eppendorf tubes and analyzed by calibrated pH meter (827 pH Lab, Metrohm, Switzerland).
  • Viscosity of the study/release medium (in this case aVH) would have significant impact on the rate of release of therapeutic agents from the DDS systems. Therefore, viscosity of the aVH has to be similar to the natural VH.
  • VH is the liquid tissue which behaves like a gel under pressure when it is in the eye. As soon as VH is isolated from the eye, it liquefies mainly due to the disorientation of collagen fibers. Rheological properties of VH are highly influenced by collagen fibrillary concentration and orientation. Usually collagen fibrils are arranged in a specific order initiating from vitreous base to the back of the eye (Le Goff et al., Adult vitreous structure and postnatal changes., Eye.
  • Freshly isolated porcine eyes were procured from a slaughter house without any further processing such as heat treatment. Throughout the transportation, the eyes were stored in isotonic solution kept in an ice bath. Prior to the viscosity measurement, the eyes were stored at 37° C. for 2 hours. After the incubation, eyes were cut open from sclera (1 ⁇ 1 cm) to access VH. Vibration plates were inserted in the eyes making sure that the plates do not touch at the bottom or side walls. Also, to understand the impact of orientation of collagen fiber on the overall viscosity, the viscosity was measured by placing vibrating plates perpendicular or parallel to collagen fibers.
  • VH As described in FIG. 2 , natural, here porcine, VH exhibited 71 ⁇ 43 mPa ⁇ S of viscosity when the plates were placed parallel to the collagen fibers. Interestingly, viscosity was not significantly different when the plates were located perpendicularly to the collagen fibers (78 ⁇ 42 mPa ⁇ S). Because hyaluronic acid along with collagen (in a defined orientation) is mainly responsible for the viscosity of the natural VH, the same components were considered as a viscosity enhancer in the aVH. According to concentration vs viscosity results of hyaluronic acid solution (Table 7), 0.6% w/v of HA was used to prepare aVH. Here, it is important to note that similar viscosity of the aVH can be achieved using different molecular weight hyaluronic acids at different concentrations.
  • aVH was prepared as described in previous section and then a defined amount of hyaluronic acid (HA) was added to prepare 0.6% of HA-SBM solution. HA was solubilized in SBM by stirring overnight at 2-8° C. The following day, collagen type-II was added in the HA-SBM solution to prepare the final collagen concentration of 40 mg/L. The resulting aVH was aseptically filtered through 0.22 ⁇ filter. The aVH was stored at 2-8° C. until further use. The final composition of a preferred aVH is reported in Table 8.
  • HA hyaluronic acid
  • aVH was investigated for the storage stability at 37° C.
  • 3 mL of aVH was aseptically transferred in 6 mL sterile glass (type-I) vial and incubated at 37° C. for 3 months.
  • Samples were collected at pre-defined time intervals i.e., initial, week-1, week-2, week-5, week-8 and week-12, and analyzed for pH, osmolality, density and viscosity. The analysis was performed with the protocols described herein.
  • aVH was found stable for 3 months at 37° C., 50 ⁇ l of 120 mg/ml bispecific monoclonal antibody (mAb) with a molecular weight of approximately 150 kDa was injected in the aVH and natural VH, and investigated for its stability. It was hypothesized that if the stability of mAb in natural VH and aVH found comparable, it can be confirmed that aVH has the potential to substitute natural VH for in-vitro/ex-vivo stability and/or release (from DDS formulations) investigations.
  • mAb bispecific monoclonal antibody
  • Porcine eyes were opened with the incision placed near the conjunctiva using a dissecting knife and the clear VH was collected with the disposable syringe without a needle.
  • the VH was then sterile-filtered through a 0.22 ⁇ m filter to ensure removal of any microbial contamination and cellular debris.
  • the VH was stored (in small aliquots) below ⁇ 70° C. to avoid possible metabolic activity and degradation or change in VH. Throughout the process of VH isolation, eye balls were kept in an ice-bath. All the experiments were performed according to the Association of Research in Vision and Ophthalmology (ARVO) statement for the use of animals in ophthalmic and vision research.
  • ARVO Association of Research in Vision and Ophthalmology
  • the ExVit is a two compartment model, VH-compartment and buffer-compartment. As demonstrated in FIG. 3 a, two compartments were separated by a diffusion controlling membrane with MWCO of 50 kDa.
  • the buffer-compartment was always filled with sterile PBS (0.01 M, pH 7.4) whereas the VH-compartment was filled with the sterile pVH or sterile aVH.
  • the devices were sealed with a sterile Teflon cap and incubated overnight at 37° C. (MaxQ-4000 incubator, Thermo Scientific). Following incubation, PBS from the buffer-compartment was replaced with fresh sterile PBS (pre-incubated at 37° C.).
  • VH-compartment 50 ⁇ L (120 mg/mL) of mAb was injected in the VH-compartment. Devices were sealed and incubated at 37° C. for the defined time intervals i.e., initial, week-2, week-4 and week-8. The VH-compartment containing pVH or aVH, without mAb, were considered as negative controls. Also, to investigate the effect of pH, temperature, and PBS on the stability of the mAb, the VH-compartment, filled with PBS, containing 50 ⁇ L of mAb was used as control. At defined time intervals, samples were aseptically transferred from VH-compartments in sterile glass vials. The performance of the model was evaluated by estimating the pH, osmolality and total protein concentration of samples. Samples were further evaluated to investigate physical stability and binding affinity of mAb.
  • Test groups A, B & C and Control groups A, B, C and D were investigated for changes in pH, osmolality and clarity following long term incubation at 37° C.
  • the pH was investigated at each time interval. Briefly, 100 ⁇ L of the samples were collected from the ExVit and vial model, from the controls (pVH alone, aVH alone, and buffer+mAb) and test articles (pVH+mAb, aVH+mAb).
  • the pH and osmolality were analyzed as described earlier. The pH and osmolality results were plotted for the time interval vs pH unit or Osmol/kg, respectively.
  • the VH is always buffered to physiological pH. Therefore, in order to mimic the long-term stability of mAb, it is very important to maintain the pH of the aVH or natural VH to physiological value. It can be clearly depicted from the data shown in FIG. 4 a, that the pH value of all the test and control samples (ExVit or Vial model) following addition of a mAb formulation remained stable throughout the study period of 2 months. Furthermore, during the entire study, osmolality of the aVH and pVH were maintained between 0.31 and 0.33 Osmol/kg ( FIG. 4 b ). It indicates that the mAb formulation was always homogeneously distributed within the aVH/pVH without altering their osmolality. In this study, aVH was found stable in the ExVit model as well as in the vial and hence protein stability investigated in the aVH can be compared with isolated pVH.
  • Samples were evaluated for the physical stability of mAb by various analytical techniques e.g., microscopy, turbidimetry, size exclusion chromatography (SEC), and capillary electrophoresis sodium dodecyl sulfate-non gel sieving (CE-SDS-NGS) as indicated in the following.
  • analytical techniques e.g., microscopy, turbidimetry, size exclusion chromatography (SEC), and capillary electrophoresis sodium dodecyl sulfate-non gel sieving (CE-SDS-NGS) as indicated in the following.
  • Proteins are generally very sensitive towards the physical, chemical and environmental stress which can easily generate soluble and insoluble particles (large aggregates).
  • To evaluate insoluble particles at each time intervals 2 mL of the sample was transferred aseptically in sterile FTU tubes. The samples were assessed microscopically (Keyence VHX-600 digital microscope) for the presence of visible particles.
  • the level of fragments and soluble aggregates of mAb were assessed using size-exclusion chromatography (SEC) coupled with a UV-visible detector.
  • SEC size-exclusion chromatography
  • the SEC method was developed in-house for the estimation of mAb. Briefly, 50 ⁇ L of samples (controls and test specimens) were injected into the separation column procured from Tosch Bioscience (TSK gel, G3000SWXL, 7.8-300 mm, 5 ⁇ ). The separation of aggregates and fragments were carried out at a flow rate of 0.5 mL/min using 0.2 M phosphate buffer (pH 7.0) as a mobile phase.
  • UV-visible detection was performed at 280 nm with a Waters-2489 detector (Water Corp. MA, USA). Analysis was executed on a Waters 2695 HPLC (Waters Corp. MA, USA) and the data was processed utilizing Empower-2 software.
  • CE-SDS-NGS Capillary Electrophoresis Sodium Dodecyl Sulfate-Non Gel Sieving
  • CE-SDS-NGS levels of soluble aggregates and fragments in the stability samples were further confirmed by the CE-SDS-NGS.
  • CE-SDS-NGS analysis was performed under non-reducing condition with the Beckman Coulter Capillary Electrophoresis System (Proteome Lab PA800). The capillary was rinsed at 70 psi with 0.1 mM NaOH (for 5 min), 0.1 mM HCl (for 1 min) and deionized water (for 1 min). The SDS MW gel buffer was loaded in the capillary at 50 psi. Non-reduced samples were then injected electrokinetically at 10 kV and analysis was carried out at 15 kV. The data was processed using 32-Karat software. The area of the main peak, aggregates, fragments, light-chain and heavy-chain were calculated, and the results were plotted as peak area (%) vs time.
  • Test-A showed minor loss in main peak at 2 month time point when compared with Test-C.
  • Test-C (Vial model)
  • mAb is exposed only to the PBS present in aVH as buffer system
  • Test-A ExVit model
  • mAb is exposed additionally to the PBS present in buffer-compartment as well. This may explain marginally higher loss in main peak observed in Test-A compared to Test-C.
  • CE-SDS-NGS analysis was performed to evaluate physical stability and to assess mainly fragmentation of mAb after incubation in the PBS and VH at 37° C.
  • Results depicted in FIG. 7 demonstrate the loss of a main peak area ( FIG. 7 a ) and concurrent increase in smaller molecular weight deglycosylated variants ( FIG. 7 b ).
  • Loss of the main peak was ca. 25% in the Control-A group which was ca. 400% higher than the loss observed in the Test-B (ca. 7%).
  • the area of the LMWS for the control sample was increased more than 300% compared to the Test-B.
  • mAb incubated in the aVH with ExVit setup showed higher loss in the main peak and increase in the LMWS compared to the mAb incubated in the aVH with vial setup (Test-C).
  • physical stability of mAb in Test-A resembles more to the Control-A and physical stability of mAb in Test-C aligns more to the Test-B. It further confirms the results observed during SEC analysis. It is important to note that the LMWS observed in the Test-A, Test-B and Test-C are of the similar type which further supports the argument of using aVH in place of natural VH to predict in-vivo protein stability.
  • the specific interaction of mAb stability samples with their target antigens was assessed using a SPR-Biacore T100/T200 instrument (GE Healthcare).
  • the dual binding SPR assay was conducted to evaluate the interactions between the mAb and its target antigens.
  • the SPR assay was performed according to European Pharmacopoeia, Osmolality, Section 2.2.35, European Directorate for the Quality of Medicine, France, France, 2017, pp. 59, which is herein incorporated by reference in its entirety.
  • Capturing anti-human Fab antibody (Human Fab Capture Kit, GE Healthcare) was immobilized on a Biacore CM5-biosensor chip via standard amine coupling to achieve a coupling density of >5000 RU. Analysis was conducted at 25° C.
  • the chip surface was regenerated by a 60 s injection of 10 mM pH 2.1 glycine solution at a flow rate of 30 ⁇ g/mL.
  • Relative binding levels of the antigens to captured mAb samples were adjusted by normalization of the mAb sample capture level relative to the reference material with a defined activity of 100%. The obtained relative activity of the individual binding domains to their respective antigens was plotted against the incubation time.
  • SM Small Molecule
  • API Active Pharmaceutical Ingredient
  • LAD Long-Acting Delivery
  • LAD implants are produced by hot-melt extrusion process, very similarly to the marketed product Ozurdex which is comprised of Dexamethasone as SM API and PLGA as matrix forming polymer.
  • the SM-loaded poly(lactic-co-glycolic acid (PLGA) implants were prepared using already established in house hot-melt extrusion protocol. Briefly, four different implants (F-1, F-2, F-3 and F-4) were prepared using hot-melt extrusion. These implants were varying in polymer type (hydrophobicity and end cap), polymer molecular weight, and extrusion temperature. Detailed composition and process parameters of implants are depicted in Table 11.
  • F-1 and F-2 at 90° C.
  • F-3 at 110° C.
  • E-beam sterilization method was performed at a standard target dose of 25kGy, at room temperature using simultaneous beam processing 10 MeV, 20 KW linear accelerators Mevex (Ottawa, CN). All the implants were then immediately stored at 2-8° C. until further use.
  • a 5 mg of above different implants (F-1 to F-4) were incubated in 2 mL of an aVH for 4 weeks at 37° C. Samples were collected after 1, 7, 14, 21 and 28 days of incubation, and analyzed by HPLC method (developed in house) for the qualitative and quantitative estimation of SM. As depicted in FIG. 9 , all the implants showed different levels of SM release after 4 weeks. Implant F-2 was comprised of fastest degrading polymer and hence it showed the fastest release compared to all other implants. Implants F-1 and F-3 were identical in composition but extruded at different temperature. F-3 exhibited significantly higher release compared to F-1.
  • aVH as a release medium was able to differentiate the implants based on their key quality feature i.e., release of API.
  • This experiment in aVH clearly enables to see differences between different types of polymers and also captures differences coming from processing such as temperature and sterilization.
  • aVH as release medium is able to discriminate between formulations that are known to, or at least expected to perform differently.
  • aVH can not only be used to investigate the stability of formulations (as highlighted in previous examples) but also can be used to investigate release of API from LAD formulations.
  • aVH as disclosed herein can better mimic the in vivo conditions with regard to API release and/or drug release from LAD implants.

Landscapes

  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Epidemiology (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Inorganic Chemistry (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Engineering & Computer Science (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Oil, Petroleum & Natural Gas (AREA)
  • Biochemistry (AREA)
  • Molecular Biology (AREA)
  • Dermatology (AREA)
  • Ophthalmology & Optometry (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
  • Medicinal Preparation (AREA)
  • Peptides Or Proteins (AREA)
  • Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
US16/855,893 2017-10-25 2020-04-22 Artificial Vitreous Humor for the Investigation of Drugs and Drug Formulations Abandoned US20200345636A1 (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
US18/298,991 US20240074972A1 (en) 2017-10-25 2023-04-11 Artificial Vitreous Humor for the Investigation of Drugs and Drug Formulations

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
EP17198210.1A EP3476384A1 (de) 2017-10-25 2017-10-25 Künstlicher glaskörper zur untersuchung von arzneimitteln und arzneimittelformulierungen
EP17198210.1 2017-10-25
PCT/EP2018/078937 WO2019081447A1 (en) 2017-10-25 2018-10-23 ARTIFICIAL GLAZED MOISTURE FOR THE STUDY OF MEDICAMENTS AND FORMULATIONS OF MEDICAMENTS

Related Parent Applications (1)

Application Number Title Priority Date Filing Date
PCT/EP2018/078937 Continuation WO2019081447A1 (en) 2017-10-25 2018-10-23 ARTIFICIAL GLAZED MOISTURE FOR THE STUDY OF MEDICAMENTS AND FORMULATIONS OF MEDICAMENTS

Related Child Applications (1)

Application Number Title Priority Date Filing Date
US18/298,991 Continuation US20240074972A1 (en) 2017-10-25 2023-04-11 Artificial Vitreous Humor for the Investigation of Drugs and Drug Formulations

Publications (1)

Publication Number Publication Date
US20200345636A1 true US20200345636A1 (en) 2020-11-05

Family

ID=60382009

Family Applications (2)

Application Number Title Priority Date Filing Date
US16/855,893 Abandoned US20200345636A1 (en) 2017-10-25 2020-04-22 Artificial Vitreous Humor for the Investigation of Drugs and Drug Formulations
US18/298,991 Pending US20240074972A1 (en) 2017-10-25 2023-04-11 Artificial Vitreous Humor for the Investigation of Drugs and Drug Formulations

Family Applications After (1)

Application Number Title Priority Date Filing Date
US18/298,991 Pending US20240074972A1 (en) 2017-10-25 2023-04-11 Artificial Vitreous Humor for the Investigation of Drugs and Drug Formulations

Country Status (5)

Country Link
US (2) US20200345636A1 (de)
EP (2) EP3476384A1 (de)
JP (1) JP7260531B2 (de)
CN (1) CN111278426B (de)
WO (1) WO2019081447A1 (de)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2023152254A1 (en) * 2022-02-11 2023-08-17 Lonza Ltd In-vitro method for simulating and analyzing behavior of an ophthalmological drug in an eye

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN113655226B (zh) * 2021-10-20 2021-12-31 首都医科大学附属北京朝阳医院 用于房水检测的校准物基质及其制备方法和应用

Family Cites Families (12)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
FR1404E (fr) 1899-09-04 1903-07-22 Jean Paul Le Grand Système de protection des bandages élastiques de roues au moyen de lames métalliques sectionnées
SE442820B (sv) * 1984-06-08 1986-02-03 Pharmacia Ab Gel av tverbunden hyaluronsyra for anvendning som glaskroppssubstitut
KR101001531B1 (ko) * 2003-02-19 2010-12-16 도꾸리쯔교세이호징 가가꾸 기쥬쯔 신꼬 기꼬 혈액 분석장치 및 혈액 분석방법
US20080129961A1 (en) * 2006-11-30 2008-06-05 Abraham Schwartz Method and apparatus for measuring quantity of a fluorochrome in a biological environment
US8623395B2 (en) * 2010-01-29 2014-01-07 Forsight Vision4, Inc. Implantable therapeutic device
AU2010208046B2 (en) * 2009-01-29 2014-10-02 Forsight Vision4, Inc. Posterior segment drug delivery
CN102049067A (zh) * 2009-11-10 2011-05-11 清华大学 一种可原位交联的高分子水凝胶玻璃体替代材料
US8197849B2 (en) * 2010-02-12 2012-06-12 National Health Research Institutes Cross-linked oxidated hyaluronic acid for use as a vitreous substitute
CN101934089B (zh) * 2010-09-01 2013-05-01 北京大学人民医院 一种可眼内注射的原位交联水凝胶在制备人工玻璃体中的应用
US8642502B2 (en) * 2010-10-11 2014-02-04 TRGel, LLC Reversible gel-forming compositions for controlled delivery of bioactive substances
WO2012147497A1 (ja) * 2011-04-27 2012-11-01 株式会社メニコン 人工硝子体材料
US8623839B2 (en) * 2011-06-30 2014-01-07 Depuy Mitek, Llc Compositions and methods for stabilized polysaccharide formulations

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
Patel et al (Supplementary Information, European Journal of Pharmaceutics and Biopharmaceutics, March 2017 (epub Nov 16, 2016), volume 112) (Year: 2017) *

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2023152254A1 (en) * 2022-02-11 2023-08-17 Lonza Ltd In-vitro method for simulating and analyzing behavior of an ophthalmological drug in an eye

Also Published As

Publication number Publication date
CN111278426A (zh) 2020-06-12
EP3476384A1 (de) 2019-05-01
JP2021500310A (ja) 2021-01-07
US20240074972A1 (en) 2024-03-07
EP3700503A1 (de) 2020-09-02
CN111278426B (zh) 2023-08-04
WO2019081447A1 (en) 2019-05-02
JP7260531B2 (ja) 2023-04-18

Similar Documents

Publication Publication Date Title
US20240074972A1 (en) Artificial Vitreous Humor for the Investigation of Drugs and Drug Formulations
CN106572998B (zh) 眼用组合物
JP7486539B2 (ja) 前房内薬物送達デポ
Zhang et al. Pharmacokinetics and tolerance study of intravitreal injection of dexamethasone-loaded nanoparticles in rabbits
CN102014957B (zh) 用于治疗耳部疾病和病况的耳用调配物
Martinez et al. In vitro and in vivo considerations associated with parenteral sustained release products: a review based upon information presented and points expressed at the 2007 Controlled Release Society Annual Meeting
CN104349769B (zh) 医用有机凝胶方法和组合物
Khan et al. Development and evaluation of a novel in situ gel of sparfloxacin for sustained ocular drug delivery: in vitro and ex vivo characterization
CN107847432A (zh) D2o稳定化的药物制剂
CN104066442A (zh) 基于半氟化烷烃的稳定蛋白质组合物
Luaces-Rodriguez et al. Intravitreal anti-VEGF drug delivery systems for age-related macular degeneration
BRPI0608978A2 (pt) terapia ocular usando agonistas de receptor alfa-2 adrenérgico possuindo taxas de depuração anterior realçadas
Sayed et al. Improved corneal bioavailability of ofloxacin: biodegradable microsphere-loaded ion-activated in situ gel delivery system
Schweizer et al. Protein–polyanion interactions for the controlled release of monoclonal antibodies
JP2018532563A (ja) 眼科用組成物
Lee et al. Subcutaneous vaccination using injectable biodegradable hydrogels for long-term immune response
EP3183554B1 (de) In-vitro-verfahren und vorrichtung zur analyse des verhaltens von substanzen in einer simulierten physiologischen umgebung
Patel et al. Prediction of intraocular antibody drug stability using ex-vivo ocular model
Stein et al. Distribution of fluorescein sodium and triamcinolone acetonide in the simulated liquefied and vitrectomized Vitreous Model with simulated eye movements
Bao et al. Exosome-loaded degradable polymeric microcapsules for the treatment of vitreoretinal diseases
ES2952213T3 (es) Composiciones y métodos para tratar enfermedades oculares
Bíró et al. New Approach in Ocular Drug Delivery: In vitro and ex vivo Investigation of Cyclodextrin-Containing, Mucoadhesive Eye Drop Formulations
Schaefer et al. Evaluation of Intracameral Pentablock copolymer Thermosensitive gel for sustained drug delivery to the anterior chamber of the eye
US10583117B2 (en) Oxo-M and 4-PPBP induction of tenogenic differentiation of perivascular tendon stem cells
Kilicarslan et al. An overview: The evaluation of formation mechanisms, preparation techniques and chemical and analytical characterization methods of the in situ forming implants

Legal Events

Date Code Title Description
AS Assignment

Owner name: F. HOFFMANN-LA ROCHE AG, SWITZERLAND

Free format text: ASSIGNMENT OF ASSIGNORS INTEREST;ASSIGNORS:ALTENBURGER, ULRIKE;PATEL, SULABH PRAVINCHANDRA;SCHWACH, GREGOIRE;REEL/FRAME:052497/0461

Effective date: 20171116

Owner name: HOFFMANN-LA ROCHE INC., NEW JERSEY

Free format text: ASSIGNMENT OF ASSIGNORS INTEREST;ASSIGNOR:F. HOFFMANN-LA ROCHE AG;REEL/FRAME:052497/0499

Effective date: 20171124

Owner name: F. HOFFMANN-LA ROCHE AG, SWITZERLAND

Free format text: ASSIGNMENT OF ASSIGNORS INTEREST;ASSIGNOR:SHENDE, PANKAJ;REEL/FRAME:052497/0490

Effective date: 20171108

STPP Information on status: patent application and granting procedure in general

Free format text: DOCKETED NEW CASE - READY FOR EXAMINATION

STPP Information on status: patent application and granting procedure in general

Free format text: NON FINAL ACTION MAILED

STPP Information on status: patent application and granting procedure in general

Free format text: RESPONSE TO NON-FINAL OFFICE ACTION ENTERED AND FORWARDED TO EXAMINER

STPP Information on status: patent application and granting procedure in general

Free format text: NON FINAL ACTION MAILED

STPP Information on status: patent application and granting procedure in general

Free format text: RESPONSE TO NON-FINAL OFFICE ACTION ENTERED AND FORWARDED TO EXAMINER

STPP Information on status: patent application and granting procedure in general

Free format text: FINAL REJECTION MAILED

STCB Information on status: application discontinuation

Free format text: ABANDONED -- FAILURE TO RESPOND TO AN OFFICE ACTION