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
  • A61K9/10—Dispersions; Emulsions
  • A61K9/127—Synthetic bilayered vehicles, e.g. liposomes or liposomes with cholesterol as the only non-phosphatidyl surfactant
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K31/00—Medicinal preparations containing organic active ingredients
  • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
  • A61K31/7028—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages
  • A61K31/7034—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages attached to a carbocyclic compound, e.g. phloridzin
  • A61K31/7036—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages attached to a carbocyclic compound, e.g. phloridzin having at least one amino group directly attached to the carbocyclic ring, e.g. streptomycin, gentamycin, amikacin, validamycin, fortimicins
• • 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
  • A61K47/08—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing oxygen, e.g. ethers, acetals, ketones, quinones, aldehydes, peroxides
  • A61K47/14—Esters of carboxylic acids, e.g. fatty acid monoglycerides, medium-chain triglycerides, parabens or PEG fatty acid esters
• • 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
  • A61K47/24—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing atoms other than carbon, hydrogen, oxygen, halogen, nitrogen or sulfur, e.g. cyclomethicone or phospholipids
• • 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
  • A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
  • A61K47/28—Steroids, e.g. cholesterol, bile acids or glycyrrhetinic acid
• • 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
  • A61K9/10—Dispersions; Emulsions
  • A61K9/127—Synthetic bilayered vehicles, e.g. liposomes or liposomes with cholesterol as the only non-phosphatidyl surfactant
  • A61K9/1277—Preparation processes; Proliposomes

Definitions

• Liposomal drug formulations enable the ability to target and enhance the uptake of active agents at specific sites of disease.
• Such formulations have been developed to treat various pulmonary disorders, including those caused by pulmonary infections, where their characteristics make them an ideal choice for the inhalation delivery of anti-infective agents.
• amikacin has been packaged in liposomes, and has been studied in multiple clinical trials in adult patients for the treatment of refractory nontub erculous mycobacterial (NTM) lung disease cause by Mycobacterium avium complex (MAC).
• NTM nontub erculous mycobacterial
• MAC Mycobacterium avium complex
• liposomes containing a relatively high amikacin to lipid ratio have been prepared at the bench scale, it is well-known that it is not a routine matter to scale up such processes to produce, at the commercial manufacturing scale, liposomal formulations where parameters such as drag concentration, amount of lipid in the formulation, lipid-to-drug ratio, captured volume, drug leakage, viscosity, and particle size are consistently maintained within specification for clinical and/or commercial use.
• the present invention addresses the need for a repeatable large-scale process for preparing liposomes containing an aminoglycoside antibiotic such as amikacin, and having a high aminoglycoside-to-lipid weight ratio (and in turn, a low lipid-to-aminoglycoside weight ratio) and superior encapsulation efficiency.
• the present invention provides a method for the large-scale manufacture of a liposomal aminoglycoside formulation comprising a lipid and an aminoglycoside (e.g., amikacin), with a high aminoglycoside loading relative to the lipid concentration (i.e., a high relative weight ratio of aminoglycoside-to-lipid).
• the lipid-to-aminoglycoside (e.g., amikacin) weight ratio (also referred to as the“L/D weight ratio”) in the liposomal suspension prepared according to method of the present invention is less than 1 : 1 upon completion of the process, for example between about 0.5: 1 and about 0.9: 1.
• the lipid-to- aminoglycoside weight ratio of the liposomal suspension prepared according to the method of the present invention is about 0.7: 1 (lipid : aminoglycoside) upon completion of the manufacturing process.
• the present invention relates to a method for the large-scale manufacture of a liposomal drug formulation comprising a lipid and aminoglycoside (e.g., amikacin), wherein the aminoglycoside is contained within the liposome with a high encapsulation efficiency (e.g., an encapsulation efficiency of at least about 40% prior to washing to remove free aminoglycoside from the formulation).
• aminoglycoside e.g., amikacin
• the method comprises mixing a first stream comprising a lipid with a second stream comprising an aminoglycoside to form a combined lipid-aminoglycoside stream.
• the combined lipid-aminoglycoside stream comprises liposomally encapsulated aminoglycoside, which in one embodiment, is formed at the intersection of the lipid stream and the aminoglycoside stream.
• the lipid-aminoglycoside stream is mixed with an aqueous saline solution in a reaction vessel (see, e.g., Figure 1).
• the aminoglycoside is present in an aqueous solution prior to the mixing step.
• the lipid is present in an alcoholic solution, e.g., an ethanolic solution, prior to the mixing step.
• the lipid comprises a phospholipid and cholesterol.
• the relative flow rate ratio of the second stream comprising aminoglycoside to the first stream comprising a lipid is about 1.5: 1 (aminoglycoside stream : lipid stream) to about 2: 1 (aminoglycoside stream : lipid stream).
• the lipid comprises dipalmitoyJphosphatidylcholine (DPPC) and cholesterol.
• DPPC dipalmitoyJphosphatidylcholine
• the aqueous saline solution is added to the reaction vessel via a third stream.
• the third stream is added to the reaction vessel at the same time as the combined lipid-aminoglycoside stream. In another embodiment, the third stream is added to the reaction vessel prior to the addition of the combined lipid-aminoglycoside stream to the reaction vessel. In another embodiment, the aqueous saline solution is at about room temperature prior to entering the reaction vessel. In one embodiment, the aqueous saline solution is about 1.5% aqueous sodium chloride.
• the present invention provides a method for the large-scale manufacture of a liposomal drug formulation comprising a lipid and aminoglycoside (e.g., amikacin), wherein the aminoglycoside is encapsulated within or comp!exed with the liposome, prior to a washing step, at an encapsulation efficiency of at least 40%.
• the weight ratio of lipid-to-aminoglycoside in the liposomal aminoglycoside formulation is less than 1 : 1, for example between about 0.5: 1 and about 0.9: 1 (e.g., about 0.7: 1).
• the aminoglycoside is amikacin.
• the amikacin is present as amikacin sulfate.
• a first stream comprising a lipid is mixed with a second stream comprising an aminoglycoside to form a combined lipid-aminoglycoside stream (e.g., a lipid- amikacin stream) comprising liposomal aminoglycoside.
• a combined lipid-aminoglycoside stream e.g., a lipid- amikacin stream
• the liposomal aminoglycoside formulation is formed at the intersection of the two streams, i.e , upon formation of the combined lipid-aminoglycoside stream.
• the flow rate of the first stream comprising a lipid is from about 0.5 kg/min to about 1.5 kg/min and the flow rate of the second stream comprising aminoglycoside is from about 1 kg/min to about 2 kg/min.
• the flow' rate of the first stream comprising a lipid is from about 3 kg/min to about 4 kg/min and the flow rate of the second stream comprising the aminoglycoside is from about 5 kg/min to about 7 kg/min.
• the relative flow' rate ratio of the second stream comprising aminoglycoside to the first stream comprising a lipid is about 1 5: 1 (aminoglycoside stream flow' rate : lipid stream flow rate) to about 2: 1 (aminoglycoside stream flow' rate : lipid stream flow rate).
• the lipid comprises dipalmitoylphosphatidyl choline (DPPC) and cholesterol.
• DPPC dipalmitoylphosphatidyl choline
• the method for the large-scale manufacture of a liposomal drug formulation comprises mixing a first stream comprising a lipid with a second stream comprising aminoglycoside to form a combined lipid-aminoglycoside stream, and adding the combined lipid- aminoglycoside stream to a vessel comprising an aqueous saline solution.
• the aqueous saline solution in one embodiment, is added to the reaction vessel via a third stream (see, e.g.. Figure 1)
• the flow rate of the first stream comprising a lipid is from about 0.5 kg/rnin to about 1.5 kg/rnin and the flow rate of the second stream comprising aminoglycoside is from about 1 kg/min to about 2 kg/min
• the flow rate of the third stream is from about 0.5 L/min and about 2.0 L/min, for example, from about 1.0 L/min to about 2.0 L/min, e.g. from about 1.0 L/min to about 1 .5 L/min, including about 1.25 L/min.
• the flow rate of the third stream is from about 3 L/min and about 6 L/min, for example, from about 4 L/min to about 6 L/min, e.g. from about 4.5 L/min to about 5.5 L/min, including about 5 L/min.
• aminoglycoside is intended to include the aminoglycoside free base and any pharmaceutically acceptable salt thereof.
• amikacin is intended to include amikacin free base and any pharmaceutically acceptable salt thereof (e.g., amikacin sulfate).
• the method for the large-scale manufacture of a liposomal aminoglycoside (e.g. amikacin) formulation comprises mixing a first stream comprising a lipid comprising a phospholipid with a second stream comprising aminoglycoside (e.g. amikacin) to form a combined lipid-aminoglycoside stream.
• the lipid-aminoglycoside stream is mixed with an aqueous saline solution in a reaction vessel.
• the phospholipid is a phosphatidylcholine.
• the phosphatidylcholine is DP PC.
• the lipid comprises a phospholipid and a sterol.
• the sterol is cholesterol.
• the lipid comprises DPPC and cholesterol.
• the method for the large-scale manufacture of a liposomal aminoglycoside formulation comprises mixing a first stream comprising a lipid with a second stream comprising aminoglycoside, wherein the first stream is mixed with the second stream to form a combined lipid-aminoglycoside stream.
• the combined lipid-aminoglycoside stream comprises liposomal aminoglycoside.
• the liposomal aminoglycoside in one embodiment, is formed upon mixing the first stream and second stream, e.g., at the intersection of the two streams.
• the combined lipid-aminoglycoside stream is added to a reaction vessel and mixed with an aqueous saline solution.
• the aminoglycoside stream and the lipid stream are each maintained at a temperature from about 30 °C to about 50 °C prior to mixing. In a further embodiment, the aminoglycoside and lipid streams are each maintained at a temperature of from about 35 °C to about 45 °C, for example from about 38 °C to about 42 °C prior to mixing. In one embodiment, the combined lipid-aminoglycoside stream is cooled upon entering the reaction vessel. In another embodiment, the combined lipid-aminoglycoside stream is cooled by the aqueous saline solution in the reaction vessel.
• the reaction vessel is maintained at a temperature from about 25 °C and about 40 °C, e.g., from about 27 °C to about 35 °C. In another embodiment, the reaction vessel is maintained at a temperature of about 30 °C. In another embodiment, the reaction vessel is maintained at a temperature of about 33 °C.
• a liposomal aminoglycoside formulation is manufactured on a large-scale according to a method provided herein.
• the concentration of aminoglycoside (e.g. amikacin) present in the liposomal drug formulation so prepared is about 10 g/L or greater, for example from about 50 g/L to about lOOg/L, including about 60 g/L to about 80 g/L and about 65 g/L to about 75 g/L (e.g., about 20 g/L, about 30 g/L, 40 about g/L, about 50 g/L, about 60 g/L, about 70 g/L or about 80 g/L).
• the concentration of lipid present in the liposomal drug formulation so prepared is from about 10 g/L to about 100 g/L, including about 20 g/L to about 80 g/L and about 40 g/L to about 60 g/L (e.g. about 50 g/L).
• the L/D ratio of a liposomal drug formulation manufactured on a large-scale according to a method provided herein is less than 1 : 1 , for example between about 0.5: 1 and about 0.8: 1 (e.g. about 0.7: 1).
• the liposomal drug formulation manufactured on a large-scale according to a method provided herein comprises liposome particles with a mean particle size (i.e. a mean diameter) of from about 200 nm to about 500 nm, for example from about 200 nm to about 400 nm (e.g. from about 250 nm to about 350 nm).
• a mean particle size i.e. a mean diameter
• FIGURES Figure 1 depicting one embodiment of the invention for preparing a liposomal aminoglycoside formulation.
• Figure 2 shows the effect of relative lipid/amikacin flow rates on the resulting L/D ratio of various liposomal amikacin formulations.
• the invention described herein relates to a method for manufacturing a liposomal aminoglycoside formulation on a large-scale.
• the method comprises mixing a first stream comprising a lipid (also referred to herein as a“lipid stream”) with a second stream comprising an aminoglycoside such as amikacin (also referred to herein as a“drug stream”) to form a combined lipid-aminoglycoside stream, and the lipid-aminoglycoside stream is mixed with an aqueous saline solution in a reaction vessel.
• the aqueous saline solution enters the reaction vessel via a third stream.
• the mixing of the lipid and drug streams is effected such that a turbulent flow results when forming the combined lipid-aminoglycoside stream.
• a turbulent flow is conveniently achieved using an appropriate T-shaped or Y-shaped infusion module for“in-line” mixing of the lipid and drug streams.
• the term“large-scale” means the use of at least about 5 kg aminoglycoside base starting material in the drug stream (calculated to at least about 5 kg aminoglycoside base if a pharmaceutically acceptable salt is used). In one embodiment, about 5 kg to about 50 kg aminoglycoside base starting material is used, for example about 5 kg to about 35 kg aminoglycoside base starting material. In one embodiment, at least about 8 kg aminoglycoside base starting material is used. In another embodiment, at least about 30 kg aminoglycoside base starting material is used. In one embodiment, the aminoglycoside is amikacin (e.g. amikacin sulfate).
• aminoglycoside used in the methods provided herein can be present as a pharmaceutically acceptable salt or as the free base.
• the aminoglycoside is amikacin, e.g., amikacin sulfate.
• the aminoglycoside is amikacin, apramycin, arbekacin, astromicin, capreomycin, dibekacin, framycetin, gentamicin, hygromycin B, isepamicin, kanamycin, neomycin, netilmicin, paromomycin, rhodestreptomycin, ribostamycin, sisomicin, spectinomycin, streptomycin, tobramycin, verdamicin, or a combination thereof.
• the aminoglycoside is AC4437, amikacin, apramycin, arbekacin, astromicin, bekanamycin, boholmycin, brulamycin, capreomycin, dibekacin, dactimicin, etimicin, framycetin, gentamicin, HI 07, hygromycin, hygromycin B, inosamycin, K- 4619, isepamicin, KA-5685, kanamycin, neomycin, netilmicin, paromomycm, plazomicin, ribostamycin, sisomicm, rhodestreptomycin, sorbistin, spectinomycin, sporaricin, streptomycin, tobramycin, verdamicin, vertilmicin, or a combination thereof.
• A“pharmaceutically acceptable salt” includes both acid and base addition salts.
• a pharmaceutically acceptable addition salt refers to those salts which retain the biological effectiveness and properties of the free bases, which are not biologically or otherwise undesirable, and which are formed with inorganic acids such as, but are not limited to, hydrochloric acid (HC1), hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid and the like, and organic acids such as, but not limited to, acetic acid, 2,2-dichloroacetic acid, adipic acid, alginic acid, ascorbic acid, aspartic acid, benzenesulfonic acid, benzoic acid, 4-acetamidobenzoic acid, camphoric acid, camphor- 10-sulfonic acid, capric acid, caproic acid, caprylic acid, carbonic acid, cinnamic acid, citric acid, cyclamic acid, dodecyl sulfuric acid, ethane- 1,2-disulfonic acid, e
• the pharmaceutically acceptable salt is HC1, TFA, lactate or acetate.
• the pharmaceutically acceptable salt is a sulfate salt, e.g., amikacin sulfate.
• “Liposomal aminoglycoside formulation” refers to a lipid-aminoglycoside formulation wherein the lipid is in the form of a liposome and the aminoglycoside is encapsulated by the liposome bilayer, or complexed with the liposome bilayer. Liposomes are completely closed lipid bilayer membranes containing an entrapped aqueous volume.
• Liposomes may be unilamellar vesicles (possessing a single membrane bilayer) or multilaraei!ar vesicles (onion-like structures characterized by multiple membrane bilayers, each separated from the next by an aqueous layer) or a combination thereof.
• the bilayer is composed of two lipid monolayers having a hydrophobic “tail” region and a hydrophilic“head” region.
• the structure of the membrane bilayer is such that the hydrophobic (nonpolar)“tails” of the lipid monolayers orient toward the center of the bilayer while the hydrophilic“heads” orient towards the aqueous phase.
• the lipid-aminoglycoside formulation is manufactured via a method comprising a two-stream infusion process.
• the method comprises mixing a first lipid stream with a second aminoglycoside stream in a T-shaped infusion module or Y-shaped infusion module.
• the terms“T-shaped infusion module,” and“Y-shaped infusion module” as used herein, refer to a T-shaped or Y-shaped chamber in which two or more streams are combined, for example, in which a lipid stream and a drug stream are combined to form a single lipid- aminoglycoside stream . See, e.g., the diagram at Figure 1. It will be appreciated that the infusion module will have a bore size appropriate for the required rate of the lipid and drug streams used. Examples of suitable bore sizes include, but are not limited to, 3/16” and 3/8”.
• the first stream comprises an alcoholic (e.g., ethanolic) lipid solution.
• the second stream comprises an aqueous aminoglycoside solution (e.g., aqueous amikacin solution).
• the first and second streams are mixed to form a combined lipid-aminoglycoside stream.
• the first and second streams each enter the infusion module and the first and second streams are mixed in the infusion module.
• the combined lipid- aminoglycoside stream exits the infusion module and subsequently enters the reaction vessel. See Figure 1.
• the combined lipid-aminoglycoside stream is mixed with an aqueous saline solution in a reaction vessel, e.g., the same reaction vessel that the combined lipid- aminoglycoside stream enters after exiting the infusion module.
• the aqueous saline solution comprises about 0.5-2% aqueous sodium chloride solution (e.g., about 1.5%).
• the saline solution is added to the reaction vessel prior to the combined lipid- aminoglycoside stream.
• the saline solution is added to the reaction vessel at or about the same time as the combined lipid-aminoglycoside stream.
• the saline solution is added to the reaction vessel via a third stream.
• the lipid-aminoglycoside formulation is manufactured via a method comprising a 3-stream infusion process.
• the third stream is added to the reaction vessel separately from the combined lipid-aminoglycoside stream.
• the combined lipid-aminoglycoside stream comprises liposomal aminoglycoside, (e.g. amikacin), wherein the encapsulation efficiency of the aminoglycoside within the liposomes (or comp!exed to the liposomes) is at least about 40%.
• Encapsulation efficiency refers to the amount of aminoglycoside encapsulated or eomplexed with liposomes prior to a filtration step, e.g., tangential flow filtration of the liposomal aminoglycoside formulation to remove free aminoglycoside.
• an encapsulation efficiency of between about 40% and about 70% can be achieved by mixing the lipid and aminoglycoside (e.g. amikacin) streams according to the method of this invention as herein described.
• the aminoglycoside stream and the lipid stream are each maintained at a temperature from about 30 °C to about 50 °C prior to mixing the two streams.
• the aminoglycoside and lipid streams are each maintained at a temperature of from about 35 °C to about 45 °C, for example from about 38 °C to about 42 °C prior to mixing.
• the combination of the lipid and aminoglycoside solutions exhibits exothermal behavior.
• the temperature of the combined lipid-aminoglycoside stream in one embodiment, is from about 40 °C to 55 °C.
• the temperature of the combined lipid-aminoglycoside stream is from about 45 °C to about 50 °C.
• the combined lipid-aminoglycoside stream is mixed with an aqueous saline solution in a reaction vessel, wherein the aqueous saline solution is maintained at room temperature prior to mixing with the combined lipid-aminoglycoside stream.
• an aqueous saline solution is added to the reaction vessel via a third stream, wherein the third stream is maintained at room temperature prior to mixing with the combined lipid-aminoglycoside stream.
• the combined lipid-aminoglycoside stream is cooled upon entering the reaction vessel.
• the combined lipid-aminoglycoside stream is cooled by the aqueous saline solution in the reaction vessel. In another embodiment, the combined lipid- aminoglycoside stream is cooled upon entering the reaction vessel. In one embodiment, the reaction vessel is maintained at a temperature from about 25 °C and about 40 °C, e.g., from about 27 C 'C to about 35 °C. In another embodiment, the reaction vessel is maintained at a temperature of about 30 °C. In another embodiment, the reaction vessel is maintained at a temperature of about
• the lipid component of the liposomal drug formulation manufactured by the method provided herein comprises electrically net neutral lipids, positively charged lipids, negatively charged lipids, or a combination thereof.
• the lipid component comprises electrically net neutral lipids.
• the lipid component consists essentially of electrically net neutral lipids.
• the lipid is DPPC and cholesterol.
• the lipids used in the manufacture of the liposomal formulations of the present invention can be synthetic, semi -synthetic or naturally-occurring lipids, including one or more of phospholipids, tocopherols, sterols, fatty acids, negatively-charged lipids and cationic lipids.
• the lipid component consists of electrically neutral lipids, e.g., a sterol and a phospholipid.
• the phospholipid is phosphatidylcholine (PC), phosphatidylglycerol (PG), phosphatidylinositol (PI), phosphatidylserine (PS), phosphatidylethanolamine (PE), phosphatidic acid (PA), soy phosphatidylcholine (SPC), soy phosphatidylglycerol (SPG), soy phosphatidylserine (SPS), soy phosphatidylinositol (SPI), soy phosphatidylethanolamine (SPE), and soy phosphatidic acid (SPA); hydrogenated egg and soya counterparts (e.g., hydrogenated egg phosphatidylcholine and hydrogenated soy phosphatidylcholine), phospholipids made up of ester linkages of fatty acids in the 2 and 3 of glycerol (PG), phosphatidylinositol (PI), phosphatidylserine (
• the lipid component of the liposom al drug formulation manufactured by the method provided herein comprises a phosphatidylcholine.
• the lipid component in the liposomal drug formulation comprises dipalmitoy!phosphatidyicho!ine (DPPC).
• the lipid component of the liposomal drug formulation comprises DPPC and a sterol, for example DPPC and cholesterol.
• the lipid consists essentially of DPPC and cholesterol, or consists of DPPC and cholesterol.
• the DPPC and cholesterol have a molar ratio in the range of from about 19: 1 (DPPC : cholesterol) to about 1 : 1 (DPPC : cholesterol), or from about 9: 1 (DPPC : cholesterol) to about 1 : 1(DPPC : cholesterol) , or from about 4: 1 (DPPC : cholesterol) to about 1 : 1 (DPPC : cholesterol), or from about 2: 1 (DPPC : cholesterol) to about 1 : 1 (DPPC : cholesterol).
• the DPPC and cholesterol have a molar ratio of about 2: 1 (DPPC : cholesterol).
• lipid components of the liposomal drug formulation manufactured by the method provided herein include, but are not limited to, dimyristoylphosphatklycholine (DMPC), dimyristoylphosphatidyJglycerol (DMPG), dipalmitoylphosphatidcholine (DPPC), dipa!mitoylphosphatidylglyceroi (DPPG), distearoylphosphatidylcholine (DSPC), distearoylphosphatidylglycerol (DSPG), dioleylphosphatidyl-ethanolamine (DOPE), mixed phospholipids such as palmitoylstearoylphosphatidyl-choline (PSPC), and single acylated phospholipids, for example, mono-oleoyl-phosphatidylethanolamine (MOPE).
• DMPC dimyristoylphosphatklycholine
• DPPC dipalmitoylphosphatidcholine
• DPPG dipa!mitoylphosphatidylg
• Examples of sterol compounds in the liposomal drug formulation manufactured by the method provided herein include, but are not limited to, cholesterol, esters of cholesterol including cholesterol hemi-succinate, salts of cholesterol including cholesterol hydrogen sulfate and cholesterol sulfate, ergosterol, esters of ergosterol including ergosterol hemi-succinate, salts of ergosterol including ergosterol hydrogen sulfate and ergosterol sulfate, lanosterol, esters of lanosterol including lanosterol hemi-succinate, salts of lanosterol including lanosterol hydrogen sulfate, lanosterol sulfate and tocopherol s.
• the tocopherols include tocopherol s, esters of tocopherol s including tocopherol hemi-succinates, salts of tocopherols including tocopherol hydrogen sulfates and tocopherol sulfates.
• the term “sterol compound” includes sterols, tocopherol s and the like. Tocopherols and their water-soluble derivatives have been used to form liposomes, see, e.g., PCX Publication No. 87/02219.
• the concentration of lipid in the first stream is from about 10 g/L to about 50 g/L, or from about 10 g/L to about 30g/L, or from about 15 g/L to about 25 g/L. In one embodiment, the concentration of lipid in the first stream is about 17g/L, about 18 g/L, about 19 g/L, about 20 g/L, about 21 g/L, about 22 g/L, about 23 g/L, about 24 g/L or about 25 g/L. In one embodiment, the concentration of lipid in the first strea is about 20 g/L
• the concentration of aminoglycoside in the second stream is from about 10 g/L to about 100 g/L; or from about 20 g/L to about 70 g/L, or from about 30 g/L, to about 60 g/L; or from about 40 g/L to about 50 g/L.
• the concentration of drug in the second stream is about 4 ' g/L, about 42 g/L, about 43 g/L, about 44 g/L, about 45 g/L, about 46 g/L, about 47 g/L, about 48 g/L, about 49 g/L or about 50 g/L.
• the concentration of aminoglycoside in the second stream is about 45 g/L.
• the aminoglycoside is amikacin.
• the pH of the aminoglycoside stream is from 6 to about 7, or from about 6.5 to about 7.0. In a further embodiment, the pH of the aminoglycoside stream is about 6.7.
• the aminoglycoside stream pH may be adjusted to the appropriate pH using a suitable base, such as an alkali or alkaline earth metal hydroxide, e.g. sodium hydroxide.
• the aqueous saline solution comprises about 0.5% sodium chloride to about 3% sodiu chloride, for example about 0.75%, about 1.0%, about 1.25%, about 1.5%, about 1.75%, about 2.0%, or about 2.5% sodium chloride. In one embodiment, the aqueous saline solution comprises about 1.5% sodium chloride.
• the flow rate of the lipid stream is from about 0 5 kg/min to about 1.5 kg/min and the flow rate of the aminoglycoside stream is from about 1 kg/min to about 2 kg/min. In a further embodiment, the flow rate of the lipid stream is from about 3 kg/min to about 4 kg/min and the flow rate of the drug stream is from about 5 kg/min to about 7 kg/min. In another embodiment, the relative flow rate ratio of the aminoglycoside stream to the lipid stream is about 1.5: 1 (aminoglycoside stream flow rate: lipid stream flow rate) to about 2: 1 (aminoglycoside stream flow rate : lipid stream flow rate).
• the flow rate of the third stream comprising aqueous saline solution is from about 0.5 L/min and about 2.0 L/min, for example, from about 1.0 L/min to about 2.0 L/min, e.g., from about 1.0 L/min to about 1.5 L/min, including about 1.25 L/rnin.
• the flow rate of the third stream comprising the aqueous saline solution is from about 3 L/min and about 6 L/min, for example, from about 4 L/min to about 6 L/min, e.g. from about 4.5 L/min to about 5.5 L/min, including about 5 L/min.
• the lipid and aminoglycoside (e.g., amikacin) solutions are both filtered, for example through one or more (e.g., two in series) about 0.2 pm filters, prior to mixing into a combined stream ( Figure 1).
• Figure 1 shows two filters in series, it should be noted that this number can be changed according to the preference of the user of the method. For example, one to five filters can be used to initially filter the lipid stream and the aminoglycoside stream.
• the aqueous saline solution (e.g., 1.5% saline solution) is also filtered, for example through one or more (e.g., two in series) about 0.2 pm filters, prior to mixing with the lipid-aminoglycoside combined stream in the reaction vessel.
• the liposomal suspension comprising liposomes formed at the intersection of the lipid and aminoglycoside streams, and/or in the combined lipid-aminoglycoside stream, is concentrated within the reaction vessel using a recirculating filtration system such as diafiltration.
• encapsulation efficiency refers to the amount of aminoglycoside encapsulated or complexed with liposomes prior to a filtration step, e.g., tangential flow filtration of the liposomal aminoglycoside formulation to remove free aminoglycoside.
• an encapsulation efficiency of between about 40% and about 70% e.g., from about 45% to about 55%) can be achieved by mixing the lipid and aminoglycoside (e.g. amikacin) streams according to the method of this invention as herein described.
• the resulting concentrated liposomal suspension is treated (i.e., “washed”’) with additional aqueous saline solution (e.g., filtered 1.5% saline solution) and subjected to further filtration using a recirculating filtration system such as diafiltration until the liposomal suspension contains an appropriate final aminoglycoside concentration and substantially all of the free aminoglycoside is removed.
• additional aqueous saline solution e.g., filtered 1.5% saline solution
• three or more washes e.g., 3, 4, 5 or 6 washes
• the concentration of aminoglycoside present in the liposomal aminoglycoside formulation manufactured on a large-scale according to a method provided herein is about 10 g/L or greater.
• aminoglycoside is present in the formulation at a concentration of about 20 g/L or greater.
• aminoglycoside is present in the formulation at a concentration of about 30 g/L or greater.
• aminoglycoside is present in the formulation at a concentration of about 40 g/L or greater.
• aminoglycoside is present in the formulation at a concentration of about 50 g/L or greater.
• aminoglycoside is present in the formulation at a concentration of about 60 g/L or greater. In a further embodiment, aminoglycoside is present in the formulation at a concentration of about 70 g/L or greater. In another embodiment, the aminoglycoside is present in the formulation at a concentration of from about 10 g/L to about 100 g/L. In a further embodiment, the aminoglycoside is amikacin. In one embodiment, the aminoglycoside is present in the formulation at a concentration of from about 50 g/L to about 100 g/L. In a further embodiment, the aminoglycoside is amikacin.
• the aminoglycoside is present in the formulation at a concentration of from about 60 g/L to about 80 g/L. In a further embodiment, the aminoglycoside is amikacin. In yet another embodiment, the aminoglycoside is present in the formulation at a concentration from about 65 g/L to about 80 g/L. In a further embodiment, the aminoglycoside is amikacin. In yet another embodiment, the aminoglycoside is present in the formulation at a concentration from about 65 g/L to about 75 g/L. In a further embodiment, the aminoglycoside is amikacin. In another embodiment, amikacin is present in the formulation at a concentration of about 70 g/L. In a further embodiment, the aminoglycoside is amikacin.
• the concentration of lipid present in the liposomal drug formulation manufactured on a large-scale according to a method provided herein is from about 10 g/L to about 100 g/L, including about 20 g/L to about 80 g/L and about 40 g/L to about 60 g/L (e.g., about 50 g/L).
• the lipid-to-aminog!ycoside weight ratio in a liposomal drug formulation manufactured on a large-scale according to a method provided herein is less than 1 : 1, for example between about 0.5: 1 (lipid : aminoglycoside) and about 0.8: 1 (lipid : aminoglycoside) (e.g., about 0.5: 1 (lipid : aminoglycoside) or 0.6: 1 (lipid : aminoglycoside) or 0.7: 1 (lipid : aminoglycoside) or 0.8: 1 (lipid : aminoglycoside)).
• the lipid-to- aminoglycoside weight ratio is about 0.7: 1 (lipid : aminoglycoside).
• the liposomal aminoglycoside formulation manufactured on a large-scale according to a method provided herein comprises liposome particles with a mean particle size (i.e. a mean diameter) of from about 200 nm to about 500 nm, for example from about 200 nm to about 400 nm (e.g. from about 250 nm to about 350 nm).
• the liposome diameter may be measured using commercially available light scattering technology, for example by quasi-elastic light scattering using a NicompTM 380 submicron particle sizer (Nicomp, Santa Barbara, California USA).
• the manufacture of liposomal amikacin sulfate was conducted using an aseptic process that involves the preparation of three sterile solution streams, mixing the lipid and amikacin sulfate streams at appropriate flow rates via a T-connector infusion module, collecting the combined lipid- amikacin sulfate streams containing liposomes with encapsulated amikacin sulfate in a sterilized diafiltration (reaction) vessel, adding a stream of 1 .5% aqueous sodium chloride at an appropriate flow rate to the diafiltration vessel, followed by diafiltration (including washing) and concentration of the resulting liposomal dispersion to form the final product.
• a) Solution Preparation Sufficient quantities of the following three solutions were prepared.
• ® Amikacin sulfate solution Amikacin sulfate in water for injection (WFI), pH adjusted with sodium hydroxide to 6.6-6.8.
• ® Lipid solution DPPC/cholesterol (2: 1 w/w) in ethanol.
• This step functions to remove the ethanol from the bulk solution and to wash away any“unentrapped” or free amikacin sulfate.
• Table 1 describes experiments (A) and (B), performed according to the general method of Example 1
• A a 3/8” T-connector infusion module is used.
• B a 3/16” T-connector infusion module is used.
• Example 2 In additional experiments, generally following the process of Example 1 , the lipid and amikacin stream (flow) rates were varied, and the resulting concentrations of lipid and amikacin in the liposomal formulations were measured. The L/D ratio for each experiment was calculated and the results presented in Figure 2. The results provide guidance for an optimal relative lipid/amikacin flow rate to achieve a preferred L/D ratio.

Landscapes

• Health & Medical Sciences (AREA)
• Chemical & Material Sciences (AREA)
• Life Sciences & Earth Sciences (AREA)
• Veterinary Medicine (AREA)
• Epidemiology (AREA)
• Medicinal Chemistry (AREA)
• Animal Behavior & Ethology (AREA)
• General Health & Medical Sciences (AREA)
• Public Health (AREA)
• Pharmacology & Pharmacy (AREA)
• Molecular Biology (AREA)
• Engineering & Computer Science (AREA)
• Chemical Kinetics & Catalysis (AREA)
• General Chemical & Material Sciences (AREA)
• Oil, Petroleum & Natural Gas (AREA)
• Dispersion Chemistry (AREA)
• Biophysics (AREA)
• Biochemistry (AREA)
• Medicinal Preparation (AREA)
• Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
• Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)

Priority Applications (10)

Applications Claiming Priority (2)

Related Child Applications (2)

Publications (1)

Family

ID=68386909

Family Applications (1)

Country Status (8)

Cited By (2)

Families Citing this family (1)

Citations (3)

Family Cites Families (356)

• 2019
  • 2019-05-02 EP EP19797021.3A patent/EP3787602A4/en active Pending
  • 2019-05-02 CN CN201980042856.6A patent/CN112584821A/zh active Pending
  • 2019-05-02 CA CA3098573A patent/CA3098573A1/en active Pending
  • 2019-05-02 KR KR1020207034487A patent/KR20210024451A/ko active Pending
  • 2019-05-02 WO PCT/US2019/030404 patent/WO2019213398A1/en not_active Ceased
  • 2019-05-02 JP JP2021510282A patent/JP7449275B2/ja active Active
  • 2019-05-02 AU AU2019262117A patent/AU2019262117C1/en active Active
  • 2019-05-02 US US17/052,070 patent/US20210113467A1/en not_active Abandoned
• 2024
  • 2024-03-01 JP JP2024031179A patent/JP2024075590A/ja active Pending
  • 2024-09-05 US US18/825,766 patent/US12521345B1/en active Active
• 2025
  • 2025-12-09 US US19/413,999 patent/US20260096989A1/en active Pending

Patent Citations (4)

Non-Patent Citations (1)

Also Published As

Similar Documents

Legal Events