US20160046582A1 - Crystals of laquinimod sodium and improved process for the manufacture thereof - Google Patents

Crystals of laquinimod sodium and improved process for the manufacture thereof Download PDF

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US20160046582A1
US20160046582A1 US14/773,683 US201414773683A US2016046582A1 US 20160046582 A1 US20160046582 A1 US 20160046582A1 US 201414773683 A US201414773683 A US 201414773683A US 2016046582 A1 US2016046582 A1 US 2016046582A1
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laquinimod
relative
methyl
pharmaceutical composition
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Anton Frenkel
Avital Laxer
Vladimir Ioffe
Karl-Erik Jansson
Ulf Tomas Fristedt
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Teva Pharmaceutical Industries Ltd
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D215/00—Heterocyclic compounds containing quinoline or hydrogenated quinoline ring systems
    • C07D215/02—Heterocyclic compounds containing quinoline or hydrogenated quinoline ring systems having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen atoms or carbon atoms directly attached to the ring nitrogen atom
    • C07D215/16—Heterocyclic compounds containing quinoline or hydrogenated quinoline ring systems having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen atoms or carbon atoms directly attached to the ring nitrogen atom with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
    • C07D215/48—Carbon atoms having three bonds to hetero atoms with at the most one bond to halogen
    • C07D215/54—Carbon atoms having three bonds to hetero atoms with at the most one bond to halogen attached in position 3
    • C07D215/56—Carbon atoms having three bonds to hetero atoms with at the most one bond to halogen attached in position 3 with oxygen atoms in position 4
    • 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
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/47—Quinolines; Isoquinolines
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
    • A61P1/04—Drugs for disorders of the alimentary tract or the digestive system for ulcers, gastritis or reflux esophagitis, e.g. antacids, inhibitors of acid secretion, mucosal protectants
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P13/00—Drugs for disorders of the urinary system
    • A61P13/12—Drugs for disorders of the urinary system of the kidneys
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P19/00—Drugs for skeletal disorders
    • A61P19/02—Drugs for skeletal disorders for joint disorders, e.g. arthritis, arthrosis
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A61P25/24—Antidepressants
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P27/00—Drugs for disorders of the senses
    • A61P27/02—Ophthalmic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A61P37/02—Immunomodulators
    • A61P37/06—Immunosuppressants, e.g. drugs for graft rejection
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C235/00—Carboxylic acid amides, the carbon skeleton of the acid part being further substituted by oxygen atoms
    • C07C235/02—Carboxylic acid amides, the carbon skeleton of the acid part being further substituted by oxygen atoms having carbon atoms of carboxamide groups bound to acyclic carbon atoms and singly-bound oxygen atoms bound to the same carbon skeleton
    • C07C235/04—Carboxylic acid amides, the carbon skeleton of the acid part being further substituted by oxygen atoms having carbon atoms of carboxamide groups bound to acyclic carbon atoms and singly-bound oxygen atoms bound to the same carbon skeleton the carbon skeleton being acyclic and saturated
    • C07C235/16—Carboxylic acid amides, the carbon skeleton of the acid part being further substituted by oxygen atoms having carbon atoms of carboxamide groups bound to acyclic carbon atoms and singly-bound oxygen atoms bound to the same carbon skeleton the carbon skeleton being acyclic and saturated having the nitrogen atom of at least one of the carboxamide groups bound to a carbon atom of a six-membered aromatic ring
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02—Column chromatography
    • G01N30/04—Preparation or injection of sample to be analysed
    • G01N30/06—Preparation
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/15—Medicinal preparations ; Physical properties thereof, e.g. dissolubility
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07B—GENERAL METHODS OF ORGANIC CHEMISTRY; APPARATUS THEREFOR
    • C07B2200/00—Indexing scheme relating to specific properties of organic compounds
    • C07B2200/13—Crystalline forms, e.g. polymorphs

Definitions

  • Laquinimod is a compound which has been shown to be effective in the acute experimental autoimmune encephalomyelitis (aEAE) model (U.S. Pat. No. 6,077,851). Its chemical name is N-ethyl-N-phenyl-1,2,-dihydro-4-hydroxy-5-chloro-1-methyl-2-oxoquinoline-3-carboxamide, and its Chemical Registry number is 248281-84-7.
  • Laquinimod is a small molecule having the following chemical structure:
  • Laquinimod sodium has high oral bioavailability and has been suggested as an oral formulation for the treatment of Multiple Sclerosis (MS).
  • MS Multiple Sclerosis
  • 2013/0217724 discloses the impurity N-ethyl-4,5-dihydroxy-1-methyl-2-oxo-N-phenyl-1,2-dihydroquinoline-3-carboxamide.
  • U.S. Patent Application Publication No. 2013/00345256 discloses the impurity N-ethyl-4-hydroxy-1-methyl-5-(methyl(2,3,4,5,6-pentahydroxyhexyl)amino)-2-oxo-N-phenyl-1,2-dihydroquinoline-3-carboxamide.
  • laquinimod sodium in U.S. Pat. No. 6,077,851, was so suspended in ethanol, and 5M sodium hydroxide solution was added. After stirring, the resulting precipitate was filtered, washed with ethanol, and dried.
  • the method used to make laquinimod sodium in U.S. Pat. No. 6,077,851 is commonly referred to as a slurry-to-slurry salt formation.
  • U.S. Pat. No. 6,875,869 discloses a process of preparing the base compound laquinimod in high yield and low level of impurities.
  • the process in U.S. Pat. No. 6,875,869 is only for synthesis of the base compound (laquinimod acid) and not the salt. As such, the slurry-to-slurry salt formation process would still be needed to form the sodium salt.
  • U.S. Pat. No. 7,884,208 teaches an improved process for preparing laquinimod sodium resulting in crystals of higher purity as well as crystals having improved crystalline characteristics, e.g., comprising no more than 2 ppm of a heavy metal and having higher tapped density.
  • laquinimod sodium is dissolved in water to form an aqueous solution; the aqueous solution is concentrated; and then a water-miscible anti-solvent is added to the concentrated solution to form laquinimod sodium crystals.
  • the process of U.S. Pat. No. 7,884,208 removes the impurities after salt formation, thus resulting in laquinimod sodium of higher purity than the laquinimod sodium produced directly from the “slurry to slurry” process of U.S. Pat. No. 6,077,851.
  • the subject invention provides a mixture of crystalline laquinimod sodium particles, wherein (i) 90% or more of the total amount by volume of the laquinimod sodium particles have a size of 40 microns or less or (ii) 50% or more of the total amount by volume of the laquinimod sodium particles have a size of 15 microns or less, and wherein:
  • the subject invention provides a mixture of crystalline laquinimod sodium particles, wherein (i) 90% or more of the total amount by volume of the laquinimod sodium particles have a size of less than 40 microns, (ii) 50% or more of the total amount by volume of the laquinimod sodium particles have a size of less than 15 microns, and (iii) 10% or more of the total amount by volume of the laquinimod sodium panicles have a size of less than 5 microns and wherein:
  • the subject invention provides a pharmaceutical composition
  • a pharmaceutical composition comprising the mixture as described herein and a pharmaceutically acceptable carrier.
  • the subject invention provides a process of recrystallization of laquinimod sodium comprising:
  • the subject invention also provides for a mixture of crystalline laquinimod sodium particles prepared by the process as described herein, and a pharmaceutical composition comprising said mixture.
  • the subject invention provides a mixture of crystalline laquinimod sodium particles, wherein (i) 90% or more of the total amount by volume of the laquinimod sodium particles have a size of 40 microns or less or (ii) 50% or more of the total amount by volume of the laquinimod sodium particles have size of 15 microns or less, and wherein
  • the subject invention also provides a pharmaceutical composition
  • a pharmaceutical composition comprising the mixture as described herein and a pharmaceutically acceptable carrier.
  • the subject invention also provides an isolated compound having the structure:
  • the subject invention also provides an composition comprising a compound having the structure:
  • the subject invention also provides a pharmaceutical composition comprising an amount of laquinimod and at least one of 2-chloro-6-(3-(ethyl(phenyl)amino)-2-hydroxy-N-methyl-3-oxopropanamido)benzoic acid (BH-3-HLAQ), 5-Chloro-4-hydroxy-1-methylquinolin-2(1H)-one (MCQ), 5-Chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid (MCQCA), Methyl 5-chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylate (MCQME), N-Ethylaniline (NEA), and Ethyl 5-chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylate (MCQEE), wherein a) 2-chloro-6-(3-(ethyl(phenyl)amino
  • the subject invention also provides a process for preparing 2-chloro-6-(3-(ethyl(phenyl)amino)-2-hydroxy-N-methyl-3-oxopropanamido)benzoic acid (BH-3-HLAQ) comprising the steps of: a) adding sodium hydroxide solution to a suspension of 5-chloro-N-ethyl-3-hydroxy-1-methyl-2,4-dioxo-N-phenyl-1,2,3,4-tetrahydroquinoline-3-carboxamide in water, b) stirring the mixture of step a) followed by addition of hydrochloric acid solution, c) extracting the aqueous solution with ethyl acetate, d) washing the organic phase with brine, e) drying the organic phase over sodium sulfate, f) filtering the suspension, g) evaporating the filtrate, h) purifying the residue by crystallization from isopropyl alcohol, i) cooling the suspension followed by filtering and
  • the subject invention also provides 2-chloro-6-(3-(ethyl(phenyl)amino)-2-hydroxy-N-methyl-3-oxopropanamido)benzoic acid (BH-1-3-HLAQ) prepared by the process described above.
  • the subject invention also provides a process for preparing 5-Chloro-4-hydroxy-1-methylquinolin-2(1H)-one (MCQ) comprising the steps of: a) heating a mixture of 5-Chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid (MCQCA) and dimethylsulfoxide, b) cooling the mixture of step a), and c) filtering the mixture of step b) and collecting the resulting filtrate.
  • MCQ 5-Chloro-4-hydroxy-1-methylquinolin-2(1H)-one
  • the subject invention also provides 5-Chloro-4-hydroxy-1-methylquinolin-2(1H)-one (MCQ) prepared by the process described above.
  • MCQ 5-Chloro-4-hydroxy-1-methylquinolin-2(1H)-one
  • the subject invention also provides a process for preparing 5-Chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid (MCQCA) comprising the steps of: a) heating a mixture of ethyl 5-chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylate (MCQEE) in a solution of hydrochloric acid in acetic acid, b) cooling the mixture of step a), c) diluting the mixture of step b) with 2-propanol and further cooling the diluted mixture, and d) filtering off the crystals resulting from step c).
  • MCQCA 5-Chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid
  • the subject invention also provides 5-Chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid (MCQCA) prepared by the process described above.
  • MCQCA 5-Chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid
  • the subject invention also provides a process for preparing Methyl 5-chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylate (MCQME) comprising the steps of: a) forming sodium dimethylmalonate by reaction of dimethylmalonate in dimethlformamide with sodium methoxide solution, b) reacting the intermediate 5-chloro-1-methyl-1h-benzo[D][1,3]oxazine-2,4-dione with sodium dimethylmalonate to form methyl 5-chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylate (MCQME) sodium salt, and c) acidifying methyl 5-chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylate (MCQME) sodium salt to methyl 5-chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxy
  • the subject invention also provides Methyl 5-chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylate (MCQME) prepared by the process described above.
  • MCQME Methyl 5-chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylate
  • the subject invention also provides a process for preparing Ethyl 5-chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylate (MCQEE) comprising the steps of: a) adding sodium hydride to a solution of 5-chloro-1-methyl-1h-benzo[D][1,3]oxazine-2,4-dione and diethyl malonate in dimethylformamide, b) heating the mixture of step a) while stirring, c) cooling the solution of step b), d) quenching the reaction mixture of step c), e) acidifying the mixture of step d), f) filtering then drying the mixture of step e), and g) crystallizing the crude product of step f) by dissolving in ethanol following by slow cooling.
  • MCQEE Ethyl 5-chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylate
  • the subject invention also provides Ethyl 5-chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylate (MCQEE) prepared by the process described above.
  • MCQEE Ethyl 5-chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylate
  • the subject invention also provides a process for testing whether a sample of laquinimod contains an undesirable impurity which comprises determining whether the sample contains a compound having the structure:
  • the subject invention also provides a process for preparing a validated pharmaceutical composition comprising laquinimod comprising: a) obtaining a batch of laquinimod; b) determining the amount of at least one of 5-Chloro-4-hydroxy-1-methylquinolin-2(1H)-one (MCQ), 5-Chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid (MCQCA), Methyl 5-chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylate (MCQME), N-Ethylaniline (NEA), and Ethyl 5-chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylate (MCQEE) in the batch using by an HPLC method; and c) preparing the pharmaceutical composition from the batch only if
  • the subject invention also provides a process for preparing a pharmaceutical composition comprising laquinimod, or for distributing a validated batch of a pharmaceutical composition comprising laquinimod, comprising a) obtaining a batch of laquinimod or of the pharmaceutical composition; b) performing stability testing with a sample of the batch; c) determining the total amount of at least one of 5-Chloro-4-hydroxy-1-methylquinolin-2(1H)-one (MCQ), 5-Chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid (MCQCA), Methyl 5-chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylate (MCQME), N-Ethylaniline (NEA), and Ethyl 5-chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylate (MCQEE) in the sample of the batch after stability testing
  • the subject invention also provides a process for validating a batch of a pharmaceutical product containing laquinimod or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier for distribution comprising a) subjecting a sample of the batch to stability testing; b) determining the amount of at least one of 2-chloro-6-(3-(ethyl(phenyl)amino)-2-hydroxy-N-methyl-3-oxopropanamido)benzoic acid (BH-3-1-HLAQ), 5-Chloro-4-hydroxy-1-methylquinolin-2(1H)-one (MCQ), 5-Chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid (MCQCA), Methyl 5-chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylate (MCQME), N-Ethylaniline (NEA), and Ethyl 5-chloro-4-hydroxy-1-methyl-2-o
  • the subject invention also provides a process for preparing a packaged pharmaceutical composition comprising laquinimod or a pharmaceutically acceptable salt thereof comprising: a) obtaining a batch of pharmaceutical composition comprising laquinimod or a pharmaceutically acceptable salt thereof; b) performing stability testing with a sample from the batch; c) determining the amount of at least one of 2-chloro-6-(3-(ethyl(phenyl)amino)-2-hydroxy-N-methyl-3-oxopropanamido)benzoic acid (BH-3-HLAQ), 5-Chloro-4-hydroxy-1-methylquinolin-2(1H)-one (MCQ), 5-Chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid (MCQCA), Methyl 5-chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylate (MCQME), N-Ethylaniline (NEA), and Ethy
  • the subject invention also provides an impurity or a salt thereof for use, as a reference standard to detect trace amounts of the impurity in a pharmaceutical composition comprising laquinimod or a pharmaceutically acceptable salt thereof, wherein the impurity is selected from the group consisting of 2-chloro-6-(3-(ethyl(phenyl)amino)-2-hydroxy-N-methyl-3-oxopropanamido)benzoic acid (BH-3-HLAQ), 5-Chloro-4-hydroxy-1-methylquinolin-2(1H)-one (MCQ), 5-Chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid (MCQCA), Methyl 5-chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylate (MCQME), N-Ethylaniline (NEA), and Ethyl 5-chloro-4-hydroxy-1-methyl-2-oxo-1,2-di
  • the subject invention also provides a method of determining the concentration of an impurity in a pharmaceutical composition comprising laquinimod, the method comprising, a) Preparing a sample solution from the pharmaceutical composition, b) Preparing a standard solution comprising the impurity, c) Preparing a resolution solution comprising laquinimod and the impurity, d) Preparing a buffer solution by dissolving ammonium acetate in water and adjusting to pH of 7.0 ⁇ 0.05 with aqueous ammonia or glacial acetic acid, e) Preparing a diluent solution comprising the buffer solution and acetonitrile, f) Preparing a blank solution comprising the diluent solution and aqueous acetonitrile, g) Injecting into the HPLC the resolution solution, the blank solution, the standard solution, and the sample solution, h) Running the HPLC using ultraviolet absorption at 240 nm and the diluent solution as the mobile phase,
  • the impurity is 5-Chloro-4-hydroxy-1-methylquinolin-2(1H)-one (MCQ), 5-Chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid (MCQCA), Methyl 5-chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylate (MCQME), N-Ethyl-4,5-dihydroxy-1-methyl-2-oxo-N-phenyl-1,2 dihydroquinoline-3-carboxamide (5-HLAQ) or Ethyl 5-chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylate (MCQEE).
  • MCQ 5-Chloro-4-hydroxy-1-methylquinolin-2(1H)-one
  • MQCA 5-Chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid
  • the subject invention also provides a method of determining the concentration of an impurity in a pharmaceutical composition comprising laquinimod, the method comprising, a) Preparing a sample solution from the pharmaceutical composition, b) Preparing a standard solution comprising the impurity, c) Preparing a resolution solution comprising laquinimod and the impurity, d) Preparing a buffer solution by dissolving ammonium acetate in water, and adjusting to pH of 7.0 ⁇ 0.05 with aqueous ammonia or glacial acetic acid, e) Preparing a blank solution comprising the buffer solution and acetonitrile, f) Injecting into the HPLC the resolution solution, the blank solution, the standard solution, and the sample solution, g) Running the HPLC using ultraviolet absorption at 240 nm and a mobile phase of a mixture of the buffer solution, and acetonitrile, h) Determining the retention time (RT) and the areas of the peaks of the impurity in
  • N-Ethylaniline N-Ethylaniline
  • SPIRO-LAQ 1H,3H-spiro[5-chloro-1-methylquinoline-2,4-dione-3,3′-[1]ethylindolin-[2]-one]
  • the subject invention also provides a method of determining the concentration of an impurity in a pharmaceutical composition comprising laquinimod and a pharmaceutically acceptable carrier, the method comprising, a) Preparing a sample solution from the pharmaceutical composition, b) Preparing a standard solution comprising the impurity, c) Preparing a Quantitation Limit (QL) solution comprising the impurity, d) Preparing a resolution solution comprising laquinimod and the impurity, e) Preparing a buffer solution by dissolving ammonium dihydrogen phosphate in water, and adjusting to pH of 7.0 ⁇ 0.10 with aqueous ammonia or phosphoric acid, f) Preparing a blank solution comprising the buffer solution and acetonitrile, g) Injecting into the HPLC the resolution solution, the blank solution, the QL solution, the standard solution and the sample solution, h) Running the HPLC using a ultraviolet absorption at 212 nm and a mobile phase of a
  • the impurity is 2-chloro-6-(3-(ethyl(phenyl)amino)-2-hydroxy-N-methyl-3-oxopropanamido)benzoic acid (BH-3-HLAQ) or N-ethyl-4-hydroxy-1-methyl-5-(2,3,4,5,6-pentahydroxyhexylamino)-2-oxo-N-phenyl-1,2-dihydroquinoline-3-carboxamide (MEG-LAQ).
  • the subject invention also provides a method of determining the concentration of an impurity in a pharmaceutical composition comprising laquinimod and a pharmaceutically acceptable carrier, the method comprising, a) Preparing a sample solution from the pharmaceutical composition, b) Preparing a standard solution comprising the impurity, c) Preparing a resolution solution comprising laquinimod and the impurity, d) Preparing a buffer solution by dissolving ammonium acetate in water, and adjusting to PH or 7.0 ⁇ 0.05 with aqueous ammonia or glacial acetic acid, e) Preparing a blank solution comprising the buffer solution and acetonitrile, f) Injecting into the HPLC the resolution solution, the blank solution, the standard solution and the sample solution, g) Running the HPLC using a ultraviolet absorption at 242 nm and the blank solution as the mobile phase, h) Determining the retention time (RT) and the areas of the peaks of the impurity in the chromatogram
  • the impurity is 5-Chloro-4-hydroxy-1-methylquinolin-2(1H)-one (MCQ), 5-Chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid (MCQCA), Methyl 5-chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylate (MCQME) or N-Ethyl-4,5-dihydroxy-1-methyl-2-oxo-N-phenyl-1,2 dihydroquinoline-3-carboxamide (5-HLAQ).
  • MCQ 5-Chloro-4-hydroxy-1-methylquinolin-2(1H)-one
  • MQCA 5-Chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid
  • MQME Methyl 5-chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylate
  • the subject invention also provides a method of determining the concentration of an impurity in a pharmaceutical composition comprising laquinimod and a pharmaceutically acceptable carrier, the method comprising, a) Preparing a sample solution from the pharmaceutical composition, b) Preparing a standard solution comprising the impurity, c) Preparing a resolution solution comprising laquinimod and the impurity, d) Preparing a blank solution comprising methanol and acetonitrile, e) Preparing a buffer solution by dissolving ammonium acetate in water, and adjusting to pH of 7.0 ⁇ 0.05 with aqueous ammonia or glacial acetic acid, f) Injecting into the HPLC the resolution solution, the blank solution, and the sample solution, g) Running the HPLC using a ultraviolet absorption at 240 nm and a mobile phase comprising acetonitrile and the buffer solution, h) Determining the retention time (RT) and the areas of the peaks of the impurity
  • N-Ethylaniline N-Ethylaniline
  • 5-Chloro-N-ethyl-3-hydroxy-1-methyl-2,4-dioxo-N-phenyl-1,2,3,4-tetrahydroquinoline-3-carboxamide 3-HLAQ
  • 1H,3H-spiro[5-chloro-1-methylquinoline-2,4-dione-3,3′-[1]ethylindolin-[2]-one] SPIRO-LAQ
  • FIG. 1 FIG. 1 from U.S. Pat. No. 7,884,208.
  • FIG. 2 FIG. 2 from U.S. Pat. No. 7,884,208.
  • FIG. 3 HPLC Data—Example chromatogram of mixture of laquinimod sodium.
  • FIG. 4 HPLC Data—Pure chromatogram of Laquinimod Sodium.
  • FIG. 5 Microscopic photograph of typical batch of crude Laquinimod Sodium at a first magnification level.
  • FIG. 6 Microscopic photograph of typical batch of crude Laquinimod Sodium at a second magnification level.
  • FIG. 7 Microscopic photograph of Batch C at a first magnification level.
  • FIG. 9 1H-NMR Spectrum of MCQ in DMSO.
  • FIG. 10 13C-NMR Spectrum of MCQ in DMSO.
  • FIG. 12 Fourier Transform-Infrared Spectrum of MCQ in KBr.
  • FIG. 13 1H-NMR Spectrum of MCQCA in D 2 O+KOH.
  • FIG. 14 13C-NMR Spectrum of MCQCA in D 2 O+KOH.
  • FIG. 15 Mass Spectrum of MCQCA (ES+ mode).
  • FIG. 16 Fourier Transform Infrared Spectrum of MCQCA.
  • FIG. 17 1H-NMR Spectrum of MCQEE in CDCl 3 .
  • FIG. 18 13C-NMR Spectrum of MCQEE in CDCl 3 .
  • FIG. 19 Mass Spectrum of MCQEE (ES + mode).
  • FIG. 20 Fourier Transform Infrared Spectrum of MCQEE.
  • FIG. 21 1H-NMR Spectrum of 5-HLAQ in DMSO.
  • FIG. 22 13C-NMR Spectrum of 5-HLAQ in DMSO.
  • FIG. 24 Fourier Transform Infrared Spectrum of 5-HLAQ.
  • FIG. 25 1H-NMR Spectrum of DELAQ in CDCl 3 .
  • FIG. 26 13C-NMR Spectrum of DELAQ in CDCl 3 .
  • FIG. 27 Mass Spectrum of DELAQ (ES+ mode).
  • FIG. 28 Fourier Transform Infrared Spectrum of DELAQ in KBr.
  • FIG. 29 1H-NMR Spectrum of 3-HLAQ in CDCl 3 .
  • FIG. 30 13C-NMR Spectrum of 3-HLAQ in CDCl 3 .
  • FIG. 31 Mass Spectrum of 3-HLAQ (ES+ mode).
  • FIG. 32 Fourier Transform Infrared Spectrum of 3-HLAQ.
  • FIG. 33 1H-NMR Spectrum of SPIRO-LAQ in DMSO.
  • FIG. 34 13C-NMR Spectrum of SPIRO-LAQ in DMSO.
  • FIG. 35 Mass Spectrum of SPIRO-LAQ (ES+ mode).
  • FIG. 36 Fourier Transform Infrared Spectrum of SPIRO-LAQ.
  • FIG. 37 1H-NMR of BH-3-HLAQ in DMSO.
  • FIG. 38 13C-NMR of BH-3-HLAQ in DMSO.
  • FIG. 39 Mass Spectrum of BH-3-HLAQ (ES+ mode).
  • FIG. 40 FT-IR Spectrum of BH-3-HLAQ.
  • FIG. 41 Resolution test chromatogram (Laquinimod Sodium Before Milling (Cryst) and Drug Substance)—Identification By Retention Time And Determination Of Assay And Polar Impurities/Degradation Products (Polar IDD) By HPLC.
  • FIG. 42 Diluent chromatogram (Laquinimod Sodium Before Milling (Cryst) and Drug Substance)—Identification By Retention Time And Determination Of Assay And Polar Impurities/Degradation Products (Polar IDD) By HPLC.
  • FIG. 43 Typical chromatogram for assay (Laquinimod Sodium Before Milling (Cryst) and Drug Substance)—Identification By Retention Time And Determination Of Assay And Polar Impurities/Degradation Products (Polar IDD) By HPLC.
  • FIG. 44 Typical chromatogram for Impurities testing (Laquinimod Sodium Before Milling (Cryst) and Drug Substance)—Identification By Retention Time And Determination Of Assay And Polar Impurities/Degradation Products (Polar IDD) By HPLC.
  • FIG. 45 Resolution test chromatogram (Laquinimod Sodium Before Milling (Cryst) and Drug Substance)—Determination Of N-Ethylaniline And Nonpolar Impurities (Non-Polar IDD) By HPLC.
  • FIG. 46 Diluent chromatogram (Laquinimod sodium before milling (Cryst) and Drug Substance) Determination Of N-Ethylaniline And Nonpolar Impurities (Non-Polar IDD) By HPLC.
  • FIG. 47 Typical sample chromatogram (Laquinimod sodium before milling (Cryst) and Drug Substance)—Determination Of N-Ethylaniline And Nonpolar Impurities (Non-Polar IDD) by HPLC.
  • FIG. 48 Resolution Solution 1 (Resolution Test) Chromatogram (Laquinimod Capsules) Identification And Determination Of Assay And Polar Impurities/Degradation Products By HPLC.
  • FIG. 49 Resolution Solution 2 Chromatogram (Laquinimod Capsules) Identification And Determination Of Assay And Polar Impurities/Degradation Products By HPLC.
  • FIG. 50 Diluent (Blank) chromatogram (Laquinimod Capsules) Identification and Determination of Assay and Polar Impurities/Degradation Products By HPLC.
  • FIG. 51 Sample chromatogram for Assay (Laquinimod Capsules) Identification and Determination of Assay and Polar Impurities/Degradation Products By HPLC.
  • FIG. 52 Sample chromatogram for determination of Impurities/Degradation products (Laquinimod Capsules) Identification and Determination of Assay and Polar Impurities/Degradation Products by HPLC.
  • FIG. 53 Resolution test chromatogram (Laquinimod Capsules) Determination of Non-Polar Impurities/Degradation Products.
  • FIG. 54 Blank (Diluent) chromatogram (Laquinimod Capsules) Determination of Non-Polar Impurities/Degradation Products.
  • FIG. 55 Sample chromatogram (Laquinimod Capsules) Determination of Non-Polar Impurities/Degradation Products.
  • FIG. 56 Typical Chromatogram of Resolution Solution (Laquinimod Capsules)—Determination of MEG-LAQ and BH-3-HLAQ by HPLC.
  • FIG. 57 Blank (Diluent 2) chromatogram (Laquinimod Capsules) Determination of MEG-LAQ and BH-3-HLAQ by HPLC.
  • FIG. 58 Chromatogram of QL Solution (Laquinimod Capsules)—Determination of MEG-LAQ and BH-3-HLAQ by HPLC.
  • FIG. 59 Chromatogram of Standard Solution (Laquinimod Capsules)—Determination of MEG-LAQ and BH-3-HLAQ by HPLC.
  • FIG. 60 Typical Sample Chromatogram (Laquinimod Capsules)—Determination of MEG-LAQ and BH-3-HLAQ by HPLC.
  • Laquinimod is a small molecule having the following chemical structure:
  • EAE Experimental Autoimmune Encephalomyelitis
  • MS Multiple Sclerosis
  • DSS Dextran Sodium Solphate
  • IRD Non-Obese Diabetic mice
  • EAN Experimental Autoimmune Neuritis
  • SLE Systemic Lupus Erythematosus
  • SLE Systemic Lupus Erythematosus
  • laquinimod results from a variety of mechanistic effects, including reduction of leukocyte infiltration into target tissues by modulation of chemokine-mediated T-cell adhesion, modulation of cytokine balance, down regulation of MHC class II resulting in alteration of antigen presentation, and effects on dendritic cells subpopulations (PCT International Application Publication No. WO2013/169746).
  • a pharmaceutically acceptable salt of laquinimod includes lithium, sodium, potassium, magnesium, calcium, manganese, copper, zinc, aluminum and iron. Salt formulations of laquinimod and the process for preparing the same are described, e.g., in U.S. Patent Application Publication No. 2005/0192315 and PCT International Application Publication No. WO 2005/074899, which are hereby incorporated by reference into this application.
  • the subject invention provides a mixture of crystalline laquinimod sodium particles, wherein (i) 90% or more of the total amount by volume of the laquinimod sodium particles have a size of 40 microns or less or (ii) 50% or more of the total amount by volume of the laquinimod sodium particles have a size of 15 microns or less, and wherein:
  • (i) 90% or more of the total amount by volume of the laquinimod sodium particles have a size of less than 40 microns or (ii) 50% or more of the total amount by volume of the laquinimod sodium particles have a size of less than 15 microns.
  • 10% or more of the total amount by volume of the laquinimod sodium particles have a size of 5 microns or less and wherein:
  • the subject invention provides a mixture of crystalline laquinimod sodium particles, wherein (i) 90% or more of the total amount by volume of the laquinimod sodium particles have a size of less than 40 microns, (ii) 50% or more of the total amount by volume of the laquinimod sodium particles have a size of less than 15 microns, and (iii) 10% or more of the total amount by volume of the laquinimod sodium particles have a size of less than 5 microns and wherein:
  • the mixture is prepared in a single batch comprising 2.5 kg or more of laquinimod sodium.
  • the laquinimod sodium particles are determined based on an unmilled sample of the mixture.
  • the size and amount by volume of laquinimod sodium particles are determined based on a milled sample of the mixture.
  • the mixture has a bulk density of 0.2 g/mL to 0.4 mL. In an embodiment of the mixture, the mixture has a tapped density of 0.40 g/mL to 0.7 g/mL.
  • an amount of aluminium in the mixture is less than 5 ppm relative to the amount by weight of laquinimod sodium. In an embodiment of the mixture, an amount of aluminium in the mixture is less than 2 ppm relative to the amount by weight of laquinimod sodium. In an embodiment of the mixture, an amount of calcium in the mixture is less than 60 ppm relative to the amount by weight of laquinimod sodium. In an embodiment of the mixture, an amount of calcium in the mixture is less than 25 ppm relative to the amount by weight of laquinimod sodium, hi an embodiment of the mixture, an amount of copper in the mixture is less than 1 ppm relative to the amount by weight of laquinimod sodium.
  • an amount of copper in the mixture is less than 0.6 ppm relative to the amount by weight of laquinimod sodium.
  • an amount of zinc in the mixture is less than 7 ppm relative to the amount by weight of laquinimod sodium.
  • an amount of zinc in the mixture is less than 4 ppm relative to the amount by weight of laquinimod sodium.
  • an amount of heavy metal in the mixture is no more than 20 ppm relative to the amount by weight of laquinimod sodium.
  • a total amount of polar impurities in the mixture is no more than 1.00% relative to the amount of laquinimod sodium as measured by HPLC.
  • an amount of 5-Chloro-4-hydroxy-1-methylquinolin-2(1H)-one (MCQ) in the mixture is no more than 0.15% relative to the amount of laquinimod sodium as measured by HPLC.
  • an amount of 5-chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid (MCQCA) in the mixture is no more than 0.15% relative to the amount of laquinimod sodium as measured by HPLC.
  • an amount of methyl 5-chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylate (MCQME) in the mixture is no more than 0.12% relative to the amount of laquinimod sodium as measured by HPLC. In another embodiment, the amount of MCQME in the mixture is no more than 0.10% relative to the amount of laquinimod sodium as measured by HPLC. In an embodiment of the mixture, an amount of ethyl 5-chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylate (MCQEE) in the mixture is no more than 0.10% relative to the amount of laquinimod sodium as measured by HPLC.
  • a total amount of non-polar impurities in the mixture is no more than 0.50% relative to the amount of laquinimod sodium as measured by HPLC.
  • an amount of N-ethylaniline (NEA) in the mixture is no more than 0.10% relative to the amount of laquinimod sodium as measured by HPLC.
  • an amount of N-ethyl-4,5-dihydroxy-1-methyl-2-oxo-N-phenyl-1,2-dihydroquinoline-3-carboxamide (5-HLAQ) in the mixture is no more than 0.10% relative to the amount of laquinimod sodium as measured by HPLC.
  • an amount of 5-chloro-4-hydroxy-1-methyl-2-oxo-N-phenyl-1,2-dihydroquinoline-3-carboxamide (DELAQ) in the mixture is no more than 0.10% relative to the amount of laquinimod sodium as measured by HPLC.
  • an amount of laquinimod acid in the mixture is no more than 1.00% relative to the amount of laquinimod sodium as measured by HPLC.
  • an amount of dimethyl malonate in the mixture is no more than 0.10% relative to the amount of laquinimod sodium as measured by HPLC.
  • an amount of diethyl malonate in the mixture is no more than 0.10% relative to the amount of laquinimod sodium as measured by HPLC.
  • an amount of dimethyl sulfate in the mixture is no more than 1 ppm relative to the amount by weight of laquinimod sodium.
  • an amount of water in the mixture is no more than 1.5% by weight relative to the amount of laquinimod sodium as measured by K.F. coulometric titration. In an embodiment of the mixture, an amount of sodium from 5.8% to 6.4% relative to the amount by weight of laquinimod sodium.
  • an amount of ethanol in the mixture is no more than 5000 ppm relative to the amount by weight of laquinimod sodium. In an embodiment of the mixture, an amount of n-heptane in the mixture is no more than 5000 ppm relative to the amount by weight of laquinimod sodium. In an embodiment of the mixture, an amount of n-octane in the mixture is no more than 2000 ppm relative to the amount by weight of laquinimod sodium. In another embodiment of the mixture, an amount of n-octane in the mixture is no more than 200 ppm relative to the amount by weight of laquinimod sodium.
  • an amount of n-octane in the mixture is no more than 20 ppm relative to the amount by weight of laquinimod sodium.
  • an amount of methanol in the mixture is no more than 3000 ppm relative to the amount by weight of laquinimod sodium.
  • an amount of acetone in the mixture is no more than 5000 ppm relative to the amount by weight of laquinimod sodium.
  • an amount of dioxane in the mixture is no more than 380 ppm relative to the amount by weight of laquinimod sodium.
  • an amount of dimethyl formamide in the mixture is no more than 880 ppm relative to the amount by weight of laquinimod sodium.
  • the recited compound is present in the mixture. In another embodiment, the recited compound is present in at least trace amounts.
  • the subject invention provides a pharmaceutical composition comprising the mixture of the subject invention and a pharmaceutically acceptable carrier.
  • a total amount of 5-Chloro-4-hydroxy-1-methylquinolin-2(1H)-one (MCQ) and 5-chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid (MCQCA) in the pharmaceutical composition is no more than 0.50% relative to the amount of laquinimod sodium as measured by HPLC.
  • a total amount of polar impurities in the pharmaceutical composition is no more than 2.00% relative to the amount of laquinimod sodium as measured by HPLC.
  • an amount of N-ethylaniline (NEA) in the pharmaceutical composition is no more than 0.50% relative to the amount of laquinimod sodium as measured by HPLC.
  • an amount of 5-chloro-N-ethyl-3-hydroxy-1-methyl-5-2,4-dioxo-N-phenyl-1,2,3,4-tetrahydroquinoline-3-carboxamide (3-HLAQ) in the pharmaceutical composition is no more than 0.50% relative to the amount of laquinimod sodium as measured by HPLC.
  • a total amount of non-polar impurities in the pharmaceutical composition is no more than 1.00% relative to the amount of laquinimod sodium as measured by HPLC.
  • an amount of N-ethyl-4-hydroxy-1-methyl-5-(methyl(2,3,4,5,6-pentahydroxyhexyl)amino)-2-oxo-N-phenyl-1,2-dihydroquinoline-3-carboxamide (MEG-LAQ) in the pharmaceutical composition is no more than 1.00% relative to the amount of laquinimod sodium as measured by HPLC.
  • an amount of water in the pharmaceutical composition is no more than 1.50% relative to the amount of laquinimod sodium as measured by K.F. coulometric titration. In an embodiment of the pharmaceutical composition, an amount of water in the pharmaceutical composition is no more than 0.80% relative to the amount of laquinimod sodium as measured by K.F. coulometric titration. In an embodiment of the pharmaceutical composition, an amount of sodium from 5.8% to 6.4% relative to the amount by weight of laquinimod sodium.
  • an amount of ethanol in the pharmaceutical composition is no more than 5000 ppm relative to the amount by weight of laquinimod sodium.
  • an amount of n-heptane in the pharmaceutical composition is no more than 5000 ppm relative to the amount by weight of laquinimod sodium.
  • an amount of n-octane in the pharmaceutical composition is no more than 2000 ppm relative to the amount by weight of laquinimod sodium.
  • an amount of n-octane in the pharmaceutical composition is no more than 200 ppm relative to the amount by weight of laquinimod sodium.
  • an amount of n-octane in the pharmaceutical composition is no more than 20 ppm relative to the amount by weight of laquinimod sodium.
  • an amount of methanol in the pharmaceutical composition is no more than 380 ppm relative to the amount by weight of laquinimod sodium.
  • an amount of acetone in the pharmaceutical composition is no more than 880 ppm relative to the amount by weight of laquinimod sodium.
  • an amount of dioxane in the pharmaceutical composition is no more than 380 ppm relative to the amount by weight of laquinimod sodium.
  • an amount of dimethyl formamide in the pharmaceutical composition is no more than 880 ppm relative to the amount by weight of laquinimod sodium.
  • the recited compound is present in the pharmaceutical composition. In another embodiment, the recited compound is present in at least trace amounts.
  • the subject invention provides a method of treating a subject afflicted with a form of multiple sclerosis, lupus nephritis, lupus arthritis, rheumatoid arthritis, a BDNF-related disorder, Crohn's disease, a GABA-related disorder, a cannabinoid receptor type 1 (CB1) mediated disorder, or an ocular inflammatory disorder comprising administering to the subject the mixture of the subject invention or the pharmaceutical composition of the subject invention so as to thereby treat the subject.
  • a form of multiple sclerosis lupus nephritis, lupus arthritis, rheumatoid arthritis, a BDNF-related disorder, Crohn's disease, a GABA-related disorder, a cannabinoid receptor type 1 (CB1) mediated disorder, or an ocular inflammatory disorder
  • the subject invention provides a method for alleviating a symptom of multiple sclerosis, lupus nephritis, lupus arthritis, rheumatoid arthritis, a BDNF-related disorder, Crohn's disease, a GABA-related disorder, a cannabinoid receptor type 1 (CB1) mediated disorder, or an ocular inflammatory disorder in a subject afflicted with a form of multiple sclerosis comprising administering to the subject the mixture of any one of the subject invention or the pharmaceutical composition of the subject invention thereby alleviating the symptom of multiple sclerosis in the subject.
  • CBD1 cannabinoid receptor type 1
  • the subject invention provides a use of the mixture or the pharmaceutical composition for the manufacture of a medicament for treating, or alleviating a symptom of, a form of multiple sclerosis, a GABA-related disorder, a cannabinoid receptor type 1 (CB1) mediated disorder, lupus nephritis, lupus arthritis, rheumatoid arthritis, a BDNF-related disorder, Crohn's disease, or an ocular inflammatory disorder.
  • CB1 cannabinoid receptor type 1
  • the subject invention provides a process of recrystallization of laquinimod sodium comprising: a) dissolving an amount of laquinimod sodium in water to form an aqueous solution; b) concentrating the aqueous solution to form a concentrated solution comprising approximately 1.7-1.8 mL of water per gram of laquinimod sodium; c) adding acetone to the concentrated solution of step b); and d) isolating recrystallized laquinimod sodium.
  • the amount of laquinimod sodium in step a) is 2.5 kg or greater. In an embodiment of the process, step a) is performed with 10-12 mL of water per gram of laquinimod sodium. In an embodiment of the process, step a) is performed with approximately 11 mL of water per gram of laquinimod sodium. In an embodiment of the process, step a) is performed by heating the aqueous solution to a temperature of 58-75° C. In an embodiment of the process, step a) is performed by heating the aqueous solution to a temperature of 60-73° C.
  • crystallization occurs after step a) and before step c).
  • crystallization is induced by rapid stirring during or after the concentrating step b). In an embodiment of the process, crystallization is induced by addition of a seed crystal during or after the concentrating step b). In an embodiment of the process, crystallization occurs without addition of a seed crystal.
  • step b) is performed under conditions appropriate to induce crystallization at the concentration of 1.7-1.8 mL of water per gram of laquinimod sodium. In an embodiment of the process, step b) is performed at 28-45° C. hi an embodiment of the process, step b) is performed at 30-40° C. In an embodiment of the process, step c) is performed with the concentrated solution at 40-55° C. In an embodiment of the process, step c) is performed with the concentrated solution at 45-50° C. In an embodiment of the process, step c) is performed with 6-12 mL of acetone per gram of laquinimod sodium.
  • step c) is performed with approximately 10 mL of acetone per gram of laquinimod sodium. In an embodiment of the process, step c) is performed over a period of 1-4 hours. In an embodiment of the process, step c) is performed over a period of 1.2-2.5 hours. In an embodiment of the process, step c) is followed by cooling the solution to a temperature no less than ⁇ 14° C. and no more than 6° C. In an embodiment of the process, step c) is followed by cooling the solution to a temperature no less than ⁇ 4° C. and no more than 4° C.
  • the solution is cooled over a period of 3-5 hours. In an embodiment of the process, the solution is cooled over a period of 3.5-4.5 hours.
  • step d) further comprises washing the recrystallized laquinimod sodium with 1-4 mL of acetone per gram of crude laquinimod sodium used in step a). In an embodiment of the process, step d) further comprises washing the recrystallized laquinimod sodium with approximately 3 mL of acetone per gram of crude laquinimod sodium used in step a). In an embodiment of the process, step d) further comprises drying the recrystallized laquinimod sodium for no less than one hour at 30-40° C. under a vacuum of no more than 50 mmHg.
  • the isolated recrystallized laquinimod sodium in step d) is a mixture of crystalline laquinimod sodium particles having a particle size distribution such that (i) 90% or more of the total amount by volume of the laquinimod sodium particles have a size of 40 microns or less, (ii) 50% or more of the total amount by volume of the laquinimod sodium particles have a size of 15 microns or less, and (iii) 10% or more of the total amount by volume of the laquinimod sodium particles have a size of less than 5 microns or less.
  • the subject invention provides a mixture of crystalline laquinimod sodium particles prepared by the process of the subject invention.
  • the mixture of crystalline laquinimod sodium particles prepared by the process of the subject invention (i) 90% or more of the total amount by volume of the laquinimod sodium particles have a size of 40 microns or less, (ii) 50% or more of the total amount by volume of the laquinimod sodium particles have a size of 15 microns or less, and (iii) 10% or more of the total amount by volume of the laquinimod sodium particles have a size of 5 microns or less.
  • the amount of n-octane in the mixture is no more than 200 ppm. In another embodiment, the amount of n-octane in the mixture is no more than 20 ppm.
  • the recited compound is present in the mixture. In another embodiment, the recited compound is present in at least trace amounts.
  • the subject invention provides a pharmaceutical composition comprising the mixture of crystalline laquinimod sodium particles prepared by the process of the subject invention, and a pharmaceutically acceptable carrier.
  • the amount of n-octane in the pharmaceutical composition is no more than 200 ppm relative to the amount by weight of laquinimod sodium. In another embodiment, the amount of n-octane in the pharmaceutical composition is no more than 20 ppm relative to the amount by weight of laquinimod sodium.
  • the subject invention provides a mixture of crystalline laquinimod sodium particles wherein (i) 90% or more of the total amount by volume of the laquinimod sodium particles have a size of 40 microns or less or (ii) 50% or more of the total amount by volume of the laquinimod sodium particles have a size of 15 microns or less, and wherein a) an amount of aluminium in the mixture is less than 5 ppm relative to the amount by weight of laquinimod sodium; b) an amount of calcium in the mixture is less than 60 ppm relative to the amount by weight of laquinimod sodium; c) an amount of copper in the mixture is less than 1 ppm relative to the amount by weight of laquinimod sodium; or d) an amount of zinc in the mixture is less than 7 ppm relative to the amount by weight of laquinimod sodium.
  • the recited compound is present in the pharmaceutical composition. In another embodiment, the recited compound is present in at least trace amounts.
  • the subject invention provides a pharmaceutical composition comprising the mixture of the subject invention and a pharmaceutically acceptable carrier.
  • 90% or more of the total amount by volume of the laquinimod sodium particles have a size of 1 micron or greater, 2 microns or greater, 3 microns or greater or 4 microns or greater. In an embodiment of the mixture, 90% of the total amount by volume of the laquinimod sodium particles have a size of greater than 1 micron, greater than 2 microns, greater than 3 microns, greater than 4 microns or greater than 5 microns.
  • the mixture has a tapped density of 0.4 g/mL to 0.7 g/mL, 0.44 g/mL to 0.7 g/mL, 0.45 g/mL to 0.7 g/mL, 0.46 g/mL to 0.7 g/mL or 0.5 g/mL to 0.7 g/mL.
  • the mixture has a bulk density of 0.2 g/mL to 0.4 g/mL, 0.2 g/mL to 0.33 g/mL, 0.2 g/mL to 0.31 g/mL.
  • the pharmaceutical composition comprises mannitol. In a further embodiment of the pharmaceutical composition, the pharmaceutical composition comprises meglumine. In a further embodiment of the pharmaceutical composition, the pharmaceutical composition comprises sodium stearyl fumarate.
  • not less than 70% of the labeled amount of laquinimod is dissolved in 30 minutes.
  • capsules of the pharmaceutical composition contain 90-110% of the labeled amount of laquinimod.
  • capsules of the pharmaceutical composition contain 95-105% of the labeled amount of laquinimod.
  • capsules of the pharmaceutical composition contain 98.0-102.0% of the labeled amount of laquinimod.
  • the pharmaceutical composition has content uniformity conforming to the U.S. Pharmacopeia. In a further embodiment of the pharmaceutical composition, the pharmaceutical composition has content uniformity conforming to European Pharmacopeia.
  • the subject invention also provides an isolated compound having the structure:
  • the subject invention also provides a composition comprising a compound having the structure:
  • the subject invention also provides a pharmaceutical composition comprising an amount of laquinimod and at least one of 2-chloro-6-(3-(ethyl(phenyl)amino)-2-hydroxy-N-methyl-3-oxopropanamido)benzoic acid (BH-3-HLAQ), 5-Chloro-4-hydroxy-1-methylquinolin-2(1H)-one (MCQ), 5-Chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid (MCQCA), Methyl 5-chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylate (MCQME), N-Ethylaniline (NEA), and Ethyl 5-chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylate (MCQEE), wherein a) 2-chloro-6-(3-(ethyl(phenyl)amino
  • a) 5-Chloro-4-hydroxy-1-methylquinolin-2(1H)-one (MCQ) is present in the pharmaceutical composition in an amount not more than 0.15%, relative to the concentration of laquinimod, based on a determination by an HPLC method
  • MCQCA 5-Chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid
  • MQCME Methyl 5-chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylate
  • N-Ethylaniline N-Ethylaniline
  • methyl 5-chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylate is present in the pharmaceutical composition in an amount not more than 0.12%, relative to the concentration of laquinimod, based on a determination by an HPLC method.
  • methyl 5-chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylate is present in the pharmaceutical composition in an amount not more than 0.1%, relative to the concentration of laquinimod, based on a determination by an HPLC method.
  • a) 5-Chloro-4-hydroxy-1-methylquinolin-2(1H)-one (MCQ) is present in the pharmaceutical composition in an amount less than 0.05%, relative to the concentration of laquinimod, based on a determination by an HPLC method
  • MCQCA 5-Chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid
  • MQCME Methyl 5-chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylate
  • N-Ethylaniline N-Ethylaniline
  • MCQCA 5-Chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid
  • a) 5-Chloro-4-hydroxy-1-methylquinolin-2(1H)-one (MCQ) is present in the pharmaceutical composition in an amount less than 0.02%, relative to the concentration of laquinimod, based on a determination by an HPLC method
  • MCQCA 5-Chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid
  • MQCME Methyl 5-chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylate
  • N-Ethylaniline N-Ethylaniline
  • 5-Chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid is present in the pharmaceutical composition in an amount less than 0.02%, relative to the concentration of laquinimod, based on a determination by an HPLC method.
  • a) 5-Chloro-4-hydroxy-1-methylquinolin-2(1H)-one (MCQ) is present in the pharmaceutical composition in an amount greater than 0.02%, and not more than 0.15%, relative to the concentration of laquinimod, based on a determination by an HPLC method
  • MCQCA 5-Chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid
  • MQCA 5-Chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid
  • Methyl 5-chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylate (MCQME) is present in the pharmaceutical composition in an amount greater than 0.02%, and not more than 0.15%, relative to the concentration of laquinimod, based on a determination by an HPLC method
  • methyl 5-chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylate is present in the pharmaceutical composition in an amount not more than 0.12%, relative to the concentration of laquinimod, based on a determination by an HPLC method.
  • methyl 5-chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylate is present in the pharmaceutical composition in an amount not more than 0.1%, relative to the concentration of laquinimod, based on a determination by an HPLC method.
  • MCQCA 5-Chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid
  • a) 5-Chloro-4-hydroxy-1-methylquinolin-2(1H)-one (MCQ) is present in the pharmaceutical composition in an amount greater than 0.05%, and not more than 0.15%, relative to the concentration of laquinimod, based on a determination by an HPLC method
  • MCQCA 5-Chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid
  • MQCA 5-Chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid
  • Methyl 5-chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylate (MCQME) is present in the pharmaceutical composition in an amount greater than 0.05%, and less than 0.15%, relative to the concentration of laquinimod
  • methyl 5-chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylate is present in the pharmaceutical composition in an amount not more than 0.12%, relative to the concentration of laquinimod, based on a determination by an HPLC method.
  • methyl 5-chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylate is present in the pharmaceutical composition in an amount not more than 0.1%, relative to the concentration of laquinimod, based on a determination by an HPLC method.
  • a) 5-Chloro-4-hydroxy-1-methylquinolin-2(1H)-one (MCQ) is present in the pharmaceutical composition in an amount greater than 0.02%, and less than 0.05%, relative to the concentration of laquinimod, based on a determination by an HPLC method
  • a) 5-Chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid (MCQCA) is present in the pharmaceutical composition in an amount greater than 0.02%, or greater than 0.03%, and less than 0.1%, relative to the concentration of laquinimod, based on a determination by an HPLC method
  • MCQME Methyl 5-chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylate
  • MCQME Methyl 5-chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylate
  • 5-Chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid is present in the pharmaceutical composition in an amount less than 0.05%, relative to the concentration of laquinimod, based on a determination by an HPLC method.
  • the composition further comprises at least one pharmaceutically acceptable carrier.
  • a) 2-chloro-6-(3-(ethyl(phenyl)amino)-2-hydroxy-N-methyl-3-oxopropanamido)benzoic acid is present in the pharmaceutical composition in an amount not more than 1.0%, relative to the concentration of laquinimod, based on a determination by an HPLC method, or b) N-Ethylaniline (NEA) is present in the pharmaceutical composition in an amount not more than 0.5%, relative to the concentration of laquinimod, based on a determination by an HPLC method, or c) the combined amount of 5-Chloro-4-hydroxy-1-methylquinolin-2(1H)-one (MCQ) and 5-Chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid (MCQCA) present in the pharmaceutical composition is not more than 0.5%, relative to the concentration of laquinimod, based on
  • N-Ethylaniline is present in the pharmaceutical composition in an amount not more than 0.1%, relative to the concentration of laquinimod, based on a determination by an HPLC method.
  • a) 2-chloro-6-(3-(ethyl(phenyl)amino)-2-hydroxy-N-methyl-3-oxopropanamido)benzoic acid (BH-3-HLAQ) is present in the pharmaceutical composition in an amount less than 0.05%, relative to the concentration of laquinimod, based on a determination by an HPLC method, or b) 5-Chloro-4-hydroxy-1-methylquinolin-2(1H)-one (MCQ) is present in the pharmaceutical composition in an amount less than 0.05%, relative to the concentration of laquinimod, based on a determination by an HPLC method, or c) 5-Chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid (MCQCA) is present in the pharmaceutical composition in an amount less than 0.5%, relative to the concentration of laquinimod, based on a determination by an HPLC method, or d) N-E
  • MCQCA 5-Chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid
  • a) 2-chloro-6-(3-(ethyl(phenyl)amino)-2-hydroxy-N-methyl-3-oxopropanamido)benzoic acid is present in the pharmaceutical composition in an amount less than 0.02%, relative to the concentration of laquinimod, based on a determination by an HPLC method, or b) 5-Chloro-4-hydroxy-1-methylquinolin-2(1H)-one (MCQ) is present in the pharmaceutical composition in an amount less than 0.02%, relative to the concentration of laquinimod, based on a determination by an HPLC method, or c) 5-Chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid (MCQCA) is present in the pharmaceutical composition in an amount less than 0.03%, relative to the concentration of laquinimod, based on a determination by an HPLC method, or d) Meth
  • 5-Chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid is present in the pharmaceutical composition in an amount less than 0.02%, relative to the concentration of laquinimod, based on a determination by an HPLC method.
  • a) 2-chloro-6-(3-(ethyl(phenyl)amino)-2-hydroxy-N-methyl-3-oxopropanamido)benzoic acid is present in the pharmaceutical composition in an amount greater than 0.02%, and not more than 1.0%, relative to the concentration of laquinimod, based on a determination by an HPLC method, or b) N-Ethylaniline (NEA) is present in the pharmaceutical composition in an greater than 0.03%, and not more than 0.5%, relative to the concentration of laquinimod, based on a determination by an HPLC method, or c) the combined amount of 5-Chloro-4-hydroxy-1-methylquinolin-2(1H)-one (MCQ) and 5-Chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid (MCQCA) present in the pharmaceutical composition is greater than 0.03%, and not
  • N-Ethylaniline is present in the pharmaceutical composition in an amount not more than 0.1%, relative to the concentration of laquinimod, based on a determination by an HPLC method.
  • a) 2-chloro-6-(3-(ethyl(phenyl)amino)-2-hydroxy-N-methyl-3-oxopropanamido)benzoic acid is present in the pharmaceutical composition in an amount greater than 0.05%, and not more than 1.0%, relative to the concentration of laquinimod, based on a determination by an HPLC method, or b) N-Ethylaniline (NEA) is present in the pharmaceutical composition in an greater than 0.1%, and not more than 0.5%, relative to the concentration of laquinimod, based on a determination by an HPLC method, or c) the combined amount of 5-Chloro-4-hydroxy-methylquinolin-2(1H)-one (MCQ) and 5-Chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid (MCQCA) present in the pharmaceutical composition is greater than 0.1%, and not more than 0.
  • MCQ 5-Chloro-4-hydroxy-methylquinolin-2
  • a) 2-chloro-6-(3-(ethyl(phenyl)amino)-2-hydroxy-N-methyl-3-oxopropanamido)benzoic acid (BH-3-HLAQ) is present in the pharmaceutical composition in an amount greater than 0.02%, and less than 0.05%, relative to the concentration of laquinimod, based on a determination by an HPLC method, or b) 5-Chloro-4-hydroxy-1-methylquinolin-2(1H)-one (MCQ) is present in the pharmaceutical composition in an amount greater than 0.02%, and less than 0.05%, relative to the concentration of laquinimod, based on a determination by an HPLC method, or c) 5-Chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid (MCQCA) is present in the pharmaceutical composition in an amount greater than 0.02%, and less than 0.5%, relative to the concentration of laquini
  • 5-Chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid is present in the pharmaceutical composition in an amount greater than 0.03%, relative to the concentration of laquinimod, based on a determination by an HPLC method.
  • 5-Chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid is present in the pharmaceutical composition in an amount less than 0.1%, relative to the concentration of laquinimod, based on a determination by an HPLC method.
  • the pharmaceutical composition as described herein further comprises laquinimod sodium salt.
  • the pharmaceutical composition is in an oral unit dosage form. In another embodiments, it is in the form of a capsule, a tablet, or a liquid suspension.
  • the oral unit dosage form comprises more than 0.3 mg laquinimod. In another embodiments, the oral unit dosage form comprises more than 0.5 mg laquinimod. In yet another embodiment, the oral unit dosage form comprises more than 0.6 mg laquinimod.
  • the subject invention also provides a process for preparing 2-chloro-6-(3-(ethyl(phenyl)amino)-2-hydroxy-N-methyl-3-oxopropanamido)benzoic acid (BH-3-HLAQ) comprising the steps of: a) adding sodium hydroxide solution to a suspension of 5-chloro-N-ethyl-3-hydroxy-1-methyl-2,4-dioxo-N-phenyl-1,2,3,4-tetrahydroquinoline-3-carboxamide in water, b) stirring the mixture of step a) followed by addition of hydrochloric acid solution, c) extracting the aqueous solution with ethyl acetate, d) washing the organic phase with brine, e) drying the organic phase over sodium sulfate, f) filtering the suspension, g) evaporating the filtrate, h) purifying the residue by crystallization from isopropyl alcohol, i) cooling the suspension followed by filtering and
  • the subject invention also provides 2-chloro-6-(3-(ethyl(phenyl)amino)-2-hydroxy-N-methyl-3-oxopropanamido)benzoic acid (BH-3-HLAQ) prepared by the process described above.
  • the subject invention also provides a process for preparing 5-Chloro-4-hydroxy-1-methylquinolin-2(1H)-one (MCQ) comprising the steps of: a) heating a mixture of 5-Chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid (MCQCA) and dimethylsulfoxide, b) cooling the mixture of step a), and c) filtering the mixture of step b) and collecting the resulting filtrate.
  • MCQ 5-Chloro-4-hydroxy-1-methylquinolin-2(1H)-one
  • the subject invention also provides 5-Chloro-4-hydroxy-1-methylquinolin-2(1H)-one (MCQ) prepared by the process described above.
  • MCQ 5-Chloro-4-hydroxy-1-methylquinolin-2(1H)-one
  • the subject invention also provides a process for preparing 5-Chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid (MCQCA) comprising the steps of: a) heating a mixture of ethyl 5-chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylate (MCQEE) in a solution of hydrochloric acid in acetic acid, b) cooling the mixture of step a), c) diluting the mixture of step b) with 2-propanol and further cooling the diluted mixture, and d) filtering off the crystals resulting from step c).
  • MCQCA 5-Chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid
  • the subject invention also provides 5-Chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid (MCQCA) prepared by the process described above.
  • MCQCA 5-Chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid
  • the subject invention also provides a process for preparing Methyl 5-chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylate (MCQME) comprising the steps of: a) forming sodium dimethylmalonate by reaction of dimethylmalonate in dimethlformamide with sodium methoxide solution, b) reacting the intermediate 5-chloro-1-methyl-1h-benzo[D][1,3]oxazine-2,4-dione with sodium dimethylmalonate to form methyl 5-chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylate (MCQME) sodium salt, and c) acidifying methyl 5-chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylate (MCQME) sodium salt to methyl 5-chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxy
  • the subject invention also provides Methyl 5-chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylate (MCQME) prepared by the process described above.
  • MCQME Methyl 5-chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylate
  • the subject invention also provides a process for preparing Ethyl 5-chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylate (MCQEE) comprising the steps of: a) adding sodium hydride to a solution of 5-chloro-1-methyl-1h-benzo[D][1,3]oxazine-2,4-dione and diethyl malonate in dimethylformamide, b) heating the mixture of step a) while stirring, c) cooling the solution of step b), d) quenching the reaction mixture of step c), e) acidifying the mixture of step d), f) filtering then drying the mixture of step e), and g) crystallizing the crude product of step f) by dissolving in ethanol following by slow cooling.
  • MCQEE Ethyl 5-chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylate
  • the subject invention also provides Ethyl 5-chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylate (MCQEE) prepared by the process described above.
  • MCQEE Ethyl 5-chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylate
  • the subject invention also provides a process for testing whether a sample of laquinimod contains an undesirable impurity which comprises determining whether the sample contains a compound having the structure:
  • the subject invention also provides a process for preparing a validated pharmaceutical composition comprising laquinimod comprising: a) obtaining a batch of laquinimod; b) determining the amount of at least one of 5-Chloro-4-hydroxy-1-methylquinolin-2(1H)-one (MCQ), 5-Chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid (MCQCA), Methyl 5-chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylate (MCQME), N-Ethylaniline (NEA), and Ethyl 5-chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylate (MCQEE) in the batch using by an HPLC method; and c) preparing the pharmaceutical composition from the batch only if
  • step c) the pharmaceutical composition is prepared from the batch only if the batch is determined to have not more than 0.12% Methyl 5-chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylate (MCQME), relative to the concentration of laquinimod.
  • step c) the pharmaceutical composition is prepared from the batch only if the batch is determined to have not more than 0.1% Methyl 5-chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylate (MCQME), relative to the concentration of laquinimod.
  • the subject invention also provides a process for preparing a pharmaceutical composition comprising laquinimod, or for distributing a validated batch of a pharmaceutical composition comprising laquinimod, comprising a) obtaining a batch of laquinimod or of the pharmaceutical composition; b) performing stability testing with a sample of the batch; c) determining the total amount of at least one of 5-Chloro-4-hydroxy-1-methylquinolin-2(1H)-one (MCQ), 5-Chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid (MCQCA), Methyl 5-chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylate (MCQME), N-Ethylaniline (NEA), and Ethyl 5-chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylate (MCQEE) in the sample of the batch after stability testing
  • the process further comprises step e) distributing the batch if in step d) the batch is validated for distribution. In another embodiments, in step d) the batch is validated for distribution or the pharmaceutical composition is prepared from the batch only if the sample of the batch after stability testing contains not more than 0.12% Methyl 5-chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylate (MCQME) relative to the concentration of laquinimod.
  • MCQME Methyl 5-chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylate
  • step d) the batch is validated for distribution or the pharmaceutical composition is prepared from the batch only if the sample of the batch after stability testing contains not more than 0.1% Methyl 5-chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylate (MCQME), relative to the concentration of laquinimod.
  • MCQME Methyl 5-chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylate
  • the subject invention also provides a process for validating a batch of a pharmaceutical product containing laquinimod or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier for distribution comprising a) subjecting a sample of the batch to stability testing; b) determining the amount of at least one of 2-chloro-6-(3-(ethyl(phenyl)amino)-2-hydroxy-N-methyl-3-oxopropanamido)benzoic acid (BH-3-HLAQ), 5-Chloro-4-hydroxy-1-methylquinolin-2(1H)-one (MCQ), 5-Chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid (MCQCA), Methyl 5-chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylate (MCQME), N-Ethylaniline (NEA), and Ethyl 5-chloro-4-hydroxy-1-methyl-2-ox
  • step c) the batch is validated for distribution only if the sample of the batch after stability testing not more than a total of the 0.1% of N-Ethylaniline (NEA) relative to the concentration of laquinimod.
  • NAA N-Ethylaniline
  • the subject invention also provides a process for preparing a packaged pharmaceutical composition comprising laquinimod or a pharmaceutically acceptable salt thereof comprising; a) a) obtaining a batch of pharmaceutical composition comprising laquinimod or a pharmaceutically acceptable salt thereof; b) performing stability testing with a sample from the batch; c) determining the amount of at least one of 2-chloro-6-(3-(ethyl(phenyl)amino)-2-hydroxy-N-methyl-3-oxopropanamido)benzoic acid (BH-3-HLAQ), 5-Chloro-4-hydroxy-1-methylquinolin-2(1H)-one (MCQ), 5-Chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid (MCQCA), Methyl 5-chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylate (MCQME), N-Ethylaniline (NEA), and
  • step d) the pharmaceutical composition is packaged only if the content of N-Ethylaniline (NEA) in the sample is determined to be not more than 0.1% to the concentration of laquinimod.
  • NAA N-Ethylaniline
  • the laquinimod is laquinimod sodium salt.
  • the subject invention also provides an impurity or a salt thereof for use, as a reference standard to detect trace amounts of the impurity in a pharmaceutical composition comprising laquinimod or a pharmaceutically acceptable salt thereof, wherein the impurity is selected from the group consisting of 2-chloro-6-(3-(ethyl(phenyl)amino)-2-hydroxy-N-methyl-3-oxopropanamido)benzoic acid (BH-3-HLAQ), 5-Chloro-4-hydroxy-1-methylquinolin-2(1H)-one (MCQ), 5-Chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid (MCQCA), Methyl 5-chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylate (MCQME), N-Ethylaniline (NEA), and Ethyl 5-chloro-4-hydroxy-1-methyl-2-oxo-1,2-di
  • the subject invention also provides a method of determining the concentration of an impurity in a pharmaceutical composition comprising laquinimod, the method comprising, a) Preparing a sample solution from the pharmaceutical composition, b) Preparing a standard solution comprising the impurity, c) Preparing a resolution solution comprising laquinimod and the impurity, d) Preparing a buffer solution by dissolving ammonium acetate in water and adjusting to pH of 7.0 ⁇ 0.05 with aqueous ammonia or glacial acetic acid, e) Preparing a diluent solution comprising the buffer solution and acetonitrile, f) Preparing a blank solution comprising the diluent solution and aqueous acetonitrile, g) Injecting into the HPLC the resolution solution, the blank solution, the standard solution, and the sample solution, h) Running the HPLC using ultraviolet absorption at 240 nm and the diluent solution as the mobile phase,
  • the impurity is 5-Chloro-4-hydroxy-1-methylquinolin-2(1H)-one (MCQ), 5-Chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid (MCQCA), Methyl 5-chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylate (MCQME). N-Ethyl-4,5-dihydroxy-1-methyl-2-oxo-N-phenyl-1,2 dihydroquinoline-3-carboxamide (5-HLAQ) or Ethyl 5-chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylate (MCQEE).
  • the subject invention also provides a method of determining the concentration of an impurity in a pharmaceutical composition comprising laquinimod, the method comprising, a) Preparing a sample solution from the pharmaceutical composition, b) Preparing a standard solution comprising the impurity, c) Preparing a resolution solution comprising laquinimod and the impurity, d) Preparing a buffer solution by dissolving ammonium acetate in water, and adjusting to pH of 7.0 ⁇ 0.05 with aqueous ammonia or glacial acetic acid, e) Preparing a blank solution comprising the buffer solution and acetonitrile, f) Injecting into the HPLC the resolution solution, the blank solution, the standard solution, and the sample solution, g) Running the HPLC using ultraviolet absorption at 240 nm and a mobile phase of a mixture of the buffer solution, and acetonitrile, h) Determining the retention time (RT) and the areas of the peaks of the impurity in
  • the impurity is N-Ethylaniline (NEA), 1H,3H-spiro[5-chloro-1-methylquinoline-2,4-dione-3,3′-[l]ethylindolin-[2]-one](SPIRO-LAQ), or 5-Chloro-N-ethyl-3-hydroxy-1-methyl-2,4-dioxo-N-phenyl-1,2,3,4-tetrahydroquinoline-3-carboxamide (3-HLAQ).
  • NAA N-Ethylaniline
  • SPIRO-LAQ 1H,3H-spiro[5-chloro-1-methylquinoline-2,4-dione-3,3′-[l]ethylindolin-[2]-one]
  • SPIRO-LAQ 5-Chloro-N-ethyl-3-hydroxy-1-methyl-2,4-dioxo-N-phenyl-1,2,3,4-tetrahydroquinoline-3-carboxamide
  • the subject invention also provides a method of determining the concentration of an impurity in a pharmaceutical composition comprising laquinimod and a pharmaceutically acceptable carrier, the method comprising, a) Preparing a sample solution from the pharmaceutical composition, b) Preparing a standard solution comprising the impurity, c) Preparing a Quantitation Limit (QL) solution comprising the impurity, d) Preparing a resolution solution comprising laquinimod and the impurity, e) Preparing a buffer solution by dissolving ammonium dihydrogen phosphate in water, and adjusting to pH of 7.0 ⁇ 0.10 with aqueous ammonia or phosphoric acid, f) Preparing a blank solution comprising the buffer solution and acetonitrile, g) Injecting into the HPLC the resolution solution, the blank solution, the QL solution, the standard solution and the sample solution, h) Running the HPLC using a ultraviolet absorption at 212 nm and a mobile phase of a
  • the impurity is 2-chloro-6-(3-(ethyl(phenyl)amino)-2-hydroxy-N-methyl-3-oxopropanamido)benzoic acid (BH-3-HLAQ) or N-ethyl-4-hydroxy-1-methyl-5-(2,3,4,5,6-pentahydroxyhexylamino)-2-oxo-N-phenyl-1,2-dihydroquinoline-3-carboxamide (MEG-LAQ).
  • the subject invention also provides a method of determining the concentration of an impurity in a pharmaceutical composition comprising laquinimod and a pharmaceutically acceptable carrier, the method comprising, a) Preparing a sample solution from the pharmaceutical composition, b) Preparing a standard solution comprising the impurity, c) Preparing a resolution solution comprising laquinimod and the impurity, d) Preparing a buffer solution by dissolving ammonium acetate in water, and adjusting to PH or 7.0 ⁇ 0.05 with aqueous ammonia or glacial acetic acid, e) Preparing a blank solution comprising the buffer solution and acetonitrile, f) Injecting into the HPLC the resolution solution, the blank solution, the standard solution and the sample solution, g) Running the HPLC using a ultraviolet absorption at 242 nm and the blank solution as the mobile phase, h) Determining the retention time (RT) and the areas of the peaks of the impurity in the chromatogram
  • the impurity is 5-Chloro-4-hydroxy-1-methylquinolin-2(1H)-one (MCQ), 5-Chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid (MCQCA), Methyl 5-chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylate (MCQME) or N-Ethyl-4,5-dihydroxy-1-methyl-2-oxo-N-phenyl-1,2 dihydroquinoline-3-carboxamide (5-HLAQ).
  • MCQ 5-Chloro-4-hydroxy-1-methylquinolin-2(1H)-one
  • MQCA 5-Chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid
  • MQME Methyl 5-chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylate
  • the subject invention also provides a method of determining the concentration of an impurity in a pharmaceutical composition comprising laquinimod and a pharmaceutically acceptable carrier, the method comprising, a) Preparing a sample solution from the pharmaceutical composition, b) Preparing a standard solution comprising the impurity, c) Preparing a resolution solution comprising laquinimod and the impurity, d) Preparing a blank solution comprising methanol and acetonitrile, e) Preparing a buffer solution by dissolving ammonium acetate in water, and adjusting to pH of 7.0 ⁇ 0.05 with aqueous ammonia or glacial acetic acid, f) Injecting into the HPLC the resolution solution, the blank solution, and the sample solution, g) Running the HPLC using a ultraviolet absorption at 240 nm and a mobile phase comprising acetonitrile and the buffer solution, h) Determining the retention time (RT) and the areas of the peaks of the impurity
  • the impurity is N-Ethylaniline (NEA), 5-Chloro-N-ethyl-3-hydroxy-1-methyl-2,4-dioxo-N-phenyl-1,2,3,4-tetrahydroquinoline-3-carboxamide (3-HLAQ) or 1H,3H-spiro[5-chloro-1-methylquinoline-2,4-dione-3,3′-[l]ethylindolin-[2]-one](SPIRO-LAQ).
  • NAA N-Ethylaniline
  • 1H,3H-spiro[5-chloro-1-methylquinoline-2,4-dione-3,3′-[l]ethylindolin-[2]-one] SPIRO-LAQ
  • any embodiment for an element of a mixture a contemplated as being applicable to a pharmaceutical composition.
  • an embodiment to a tapped density of 0.46 g/mL to 0.7 g/mL is contemplated as being applicable to a mixture having an amount of aluminium in the mixture less than 5 ppm relative to the amount by weight of laquinimod sodium.
  • U.S. Pat. No. 7,884,208 teaches a process for recrystallizing laquinimod sodium which produces a mixture of crystalline laquinimod sodium having larger crystals, lowered impurity levels and certain improved crystalline characteristics as compared to the laquinimod sodium crystals known at the time.
  • U.S. Pat. No. 7,884,208 achieves a mixture of crystalline laquinimod sodium having (i) a mixture wherein 10% or more of the total amount by volume of the laquinimod sodium particles has a size of greater than 40 microns and wherein 50% or more of the total amount by volume of the laquinimod sodium particles has a size of greater than 15 microns, (ii) high density (tapped and bulk), (iii) low heavy metal content, and (iv) low content of certain polar impurities.
  • the recrystallization process of U.S. Pat. No. 7,884,208 does not produce a mixture of recystallized laquinimod sodium particles wherein 90% or more of the total amount by volume of the laquinimod sodium have a size of less than 40 microns, wherein 50% or more of the total amount by volume of the laquinimod sodium has a size of less than 15 microns, and wherein 10% or more of the total amount by volume of the laquinimod sodium has a size of less than 5 microns.
  • Example 14 of U.S. Pat. No. 7,884,208 produced a mixture of recystallized laquinimod sodium particles wherein 10% or more of the total amount by volume of the laquinimod sodium have a size of less than 5 microns. However, this Example also shows reduced quality of crystalline characteristics, specifically Tapped Density. The mixture produced by Example 14 has an acceptable D(0.1) value but an undesired Tapped Density.
  • Example 13 of U.S. Pat. No. 7,884,208 produced a mixture having high Tapped Density, but did not produce crystals wherein 90% or more of the total amount by volume of the laquinimod sodium particles have a size of 40 microns or less or 50% or more of the total amount by volume of the laquinimod sodium particles have a size of 15 microns or less.
  • U.S. Pat. No. 7,884,208 is unable to achieve the advantages of recrystallization, i.e., better density and impurity profiles, while also producing laquinimod sodium crystals wherein 90% or more of the total amount by volume of the laquinimod sodium particles have a size of 40 microns or less, or 50% or more of the total amount by volume of the laquinimod sodium particles have a size of 15 microns or less.
  • 6,077,851 achieves a mixture wherein 90% or more of the total amount by volume of the laquinimod sodium particles have a size of 40 microns or less or, 50% or more of the total amount by volume of the laquinimod sodium particles have a size of 15 microns or less, but it does not achieve crystals having acceptable density or low levels of impurities.
  • the prior art contains no teaching of a process for preparing laquinimod sodium wherein 90% or more of the total amount by volume of the laquinimod sodium particles have a size of 40 microns or less, or 50% or more of the total amount by volume of the laquinimod sodium particles have a size of 15 microns or less, and having desirable density and purity profiles.
  • the present invention provides a process which is capable of producing a mixture of recystallized laquinimod sodium crystals wherein 90% or more of the total amount by volume of the laquinimod sodium particles have a size of 40 microns or less or (ii) 50% or more of the total amount by volume of the laquinimod sodium particles have a size of 15 microns or less, and having desirable density and purity profiles, specifically, (i) high density (tapped or bulk), (ii) low heavy metal content, or (iii) low polar impurity content.
  • the mixture of laquinimod sodium provided by the present invention is achieved by an improved recrystallization process.
  • the laquinimod sodium manufactured by the recrystallization processes of the present invention has improved purity and density profiles over the laquinimod sodium disclosed in U.S. Pat. No. 6,077,851 and improved crystalline characteristics, especially smaller particles, over U.S. Pat. No. 7,884,208.
  • the modified recrystallization process of the present invention unexpectedly results in different recrystallization conditions than achieved by the process disclosed in U.S. Pat. No. 7,884,208 and, thusly, results in different products.
  • concentrating the aqueous solution to 1.7-1.8 unexpectedly results in crystalline laquinimod sodium particles having reduced levels of impurities, improved crystalline characteristics, and wherein 90% or more of the total amount by volume of the laquinimod sodium particles have a size of 40 microns or less, 50% or more of the total amount by volume of the laquinimod sodium particles have a size of 15 microns or less, and 10% or more of the total amount by volume of the laquinimod sodium particles have a size of 5 microns or less.
  • an important factor affecting crystallization is initial concentration of crystallizing solution. Careful control of concentration of the solution can facilitate initiation of spontaneous crystallization prior to the end of the concentration step.
  • the process of the present invention can initiate spontaneous crystallization prior to addition of acetone.
  • Concentrating the aqueous solution to form a concentrated solution comprising approximately 1.7-1.8 ml of water per gram of laquinimod sodium is an important aspect of the present invention.
  • U.S. Pat. No. 7,884,208 teaches advantages associated with large particles. Specifically, U.S. Pat. No. 7,884,208 teaches that larger particles of laquinimod sodium are more “processable” when making pharmaceutical compositions and that smaller particles are often associated with dust-like properties which may interfere with processing, and sometimes associated with flowability problems which may interfere with manufacturing. Further, U.S. Pat. No. 7,884,208 teaches that chemical stability has been shown to be decreased by the increase in surface area that results from smaller particle size. (Felffle, A. Chpt 88 , Remington's Pharmaceutical Sciences, 15 th Edition, Mack Publishing Company, Easton, Pa. (1975)).
  • the subject invention has unexpectedly achieved an improved mixture of laquinimod sodium wherein (i) 90% or more of the total amount by volume of the laquinimod sodium particles have a size of 40 microns or less, (ii) 50% or more of the total amount by volume of the laquinimod sodium particles have a size of 15 microns or less, and (iii) 10% or more of the total amount by volume of the laquinimod sodium particles have a size of 5 microns or less.
  • Laquinimod has been graded as a highly potent API, demanding special caution and avoiding material contact with workers and environment.
  • Laquinimod has been graded as having the highest potency rate, corresponding to a recommended acceptable daily intake (ADI) during operations and manufacturing of less than 0.01 mg/day or ⁇ 1 ⁇ g/m 3 as an 8-hour TWA.
  • High potency compounds are associated with controls, whether engineering, administrative or procedure-related, that afford the desired level of worker protection. For example, high potency compounds may require no human intervention or manual operations. (Bruce D. Naumann, Control Banding In The Pharmaceutical Industry, http://www.aioh.org.au/downloads/documents/ControlBandingBNaumaun.pdf)
  • the mixture of the subject invention may be milled or unmilled, the present invention is achieved without the need of a milling operation.
  • the mixture of laquinimod sodium of the present invention overcomes the potential problems associated with mixtures having large particles.
  • the small size of the laquinimod sodium particles of the present invention may obviate the need for milling and comminution steps.
  • U.S. Pat. No. 7,884,208 reflected the understanding in the art that there are problems associated with small particles in pharmaceutical compositions, but the laquinimod sodium of the present invention has no problems associated with stability, processing or manufacturing.
  • Comminution introduces its own set of problems to a drug substance beyond the disadvantages of introducing an additional drug processing step.
  • milling can introduce impurities, new polymorphs, amorphous sections in the crystalline structure of the API, other changes to particle morphology, differences in agglomeration, increased solubility, changes in moisture levels, and changes in compressibility (Hausner, “The Role of Particle Size in the Development of Generic Products” 2003).
  • comminution may affect the efficacy and safety of a drug substance.
  • Some of the disadvantages of comminution are illustrated by the side effects resulting from microcrystalline Nitrofurantoin compared to macrocrystalline Nitrofurantoin. (Brumfitt, W. and J. M. T. Hamilton-Miller, J. Antimicrobial Chemotherapy 42:363:371 (1998)).
  • the laquinimod sodium of the present invention provides a mixture of laquinimod sodium particles having small particle sizes which avoids safety problems and additional problems related to milling.
  • laquinimod Another concern during formulation processes is maintaining uniformity of content of the drug product.
  • the unit dose of laquinimod is quite low relative to the total weight of the drug product, e.g., tablet or capsule.
  • a typical formulation may comprise only a small amount of laquinimod, e.g., 0.3, 0.6 or 1.2 mg, in a capsule with total weight of over 200 mg.
  • small fluctuation in the amount of laquinimod due to problems of flowability, segregation, uniformity, or poor homogenous distribution could result in a large percent deviation from the desired amount, e.g., 0.3, 0.6 or 1.2 mg.
  • the mixture of laquinimod sodium of the present invention provides high uniformity of content and minimal fluctuations in the amount of laquinimod in the capsules.
  • Uniformity of the shape of laquinimod particles is also an important concern during formulation as a lack of uniformity of shape can cause variation in density of drug substance and cause problems during drug product formation, e.g., capsule or tablet formation.
  • Crystalline laquinimod sodium particles are rod-shaped particles. It is known that milling operations may result in changes to particle shape.
  • Decreased particle size is known to result in faster dissolution profiles.
  • the rate of dissolution of small particles is usually faster than that of large particles because a greater surface area of the drug substance is in contact with the liquid medium.
  • an advantage of the recrystallization process of the present invention is that the resulting mixture of crystalline laquinimod sodium has particles having small particle sizes, which is associated with high uniformity and homogeneity with respect to distribution of the API into capsules, tablets and other drug products.
  • Laquinimod sodium crystals having small particle sizes can obviate or reduce the need for additional milling steps.
  • the small particle sizes of the laquinimod sodium of the present invention are achieved without sacrificing desirable purity or density profiles and without the need for prior milling operations.
  • laquinimod sodium crystals have a higher density than the laquinimod sodium crystals produced by the slurry-to-slurry process of U.S. Pat. No. 6,077,851.
  • Low tapped density is anathema to certain prized qualities in a drug substance or drug product such as compressibility, the ability of a powder to decrease in volume under pressure, and compactability, the ability of a powder to be compressed into a tablet of certain strength or hardness. Crystals with low tapped density are also known to have poor flowability, which results in a lack of uniformity of content in finished dosage forms, especially in tablets. (Rudnic et al. Chpt.
  • Uniformity of content is especially important for pharmaceutical compositions comprising a potent drug substance, e.g., Laquinimod sodium.
  • the present invention Compared to the slurry-to-slurry process of U.S. Pat. No. 6,077,851, the present invention also shows low aggregation of the particles and, additionally, provides particles with acceptable density and lower levels of impurities. As shown in FIGS. 5-8 , the crude laquinimod resulting from the process described in U.S. Pat. No. 6,077,851 has a high rate of aggregates ( FIGS. 5 and 6 ), compared to a low rate of aggregates of the present invention ( FIGS. 7 and 8 ).
  • Another advantage of the present invention is that the process of the present invention is environmentally friendly without sacrificing desirable crystalline characteristics. Specifically, by use of water as the primary solvent, the present invention achieves both environmental friendliness and improved crystalline characteristics, specifically with respect to particle size distribution over U.S. Pat. No. 7,884,208.
  • laquinimod means laquinimod acid or a pharmaceutically acceptable salt thereof, including laquinimod sodium.
  • laquinimod acid is N-ethyl-N-phenyl-1,2,-dihydro-4-hydroxy-5-chloro-1-methyl-2-oxoquinoline-3-carboxamide, and its Chemical Registry number is 248281-84-7.
  • Laquinimod sodium is the sodium salt of laquinimod acid.
  • D(0.1) is the particle size, in microns, below which 10% by volume distribution of the population is found.
  • D(0.5) is the particle size, in microns, below which 50% by volume distribution of the population is found.
  • D(0.9) is the particle size, in microns, below which 90% by volume distribution of the population is found.
  • crystalline characteristics includes particle size distribution, bulk density and tapped density.
  • drug substance refers to the active ingredient in a drug product or for use in a drug product, which provides pharmacological activity or other direct effect in the diagnosis, cure, mitigation, treatment, or prevention of disease, or to affect the structure or any function of the body of man or animals.
  • drug product refers to the formulated or finished dosage form containing the drug substance as well as at least one pharmaceutically acceptable carrier.
  • composition that is “free” of a chemical entity means that the composition contains, if at all, an amount of the chemical entity which cannot be avoided following an affirmative act intended to purify the composition by separating the chemical entity from the composition.
  • a composition which is “free” of an alkalizing agent means that the alkalizing agent, if present at all, is a minority component of the composition by weight.
  • the composition comprises less than 0.1 wt %, 0.05 wt %, 0.02 wt %, or 0.01 wt % of the component.
  • an “isolated” compound is a compound isolated from the crude reaction mixture following an affirmative act of isolation.
  • the act of isolation involves separating the compound from the other known components of the crude reaction mixture, with some impurities, unknown side products and residual amounts of the other known components of the crude reaction mixture permitted to remain. Purification is an example of an affirmative act of isolation.
  • stability testing refers to tests conducted at specific time intervals and various environmental conditions (e.g., temperature and humidity) to see if and to what extent a drug product degrades over its designated shelf life time.
  • the specific conditions and time of the tests are such that they accelerate the conditions the drug product is expected to encounter over its shelf life.
  • detailed requirements of stability testing for finished pharmaceuticals are codified in 21 C.F.R ⁇ 211.166, the entire content of which is hereby incorporated by reference.
  • dissolution rate is determined based on the amount of drug substance dissolved in 30 min. as indicated in the U.S. Pharmacopeia ⁇ 711>.
  • atmospheric pressure refers to a pressure of about 1 atm.
  • ambient temperature refers to a temperature of about 20° C. to about 30° C.
  • stable pharmaceutical composition as used herein in connection with the composition according to the invention denotes a composition, which preserves the physical stability/integrity and/or chemical stability/integrity of the active pharmaceutical ingredient during storage.
  • stable pharmaceutical composition is characterized by its level of degradation products not exceeding 5% at 40° C./75% RH after 6 months or 3% at 55° C./75% RH after two weeks, compared to their level in time zero.
  • treating encompasses, e.g., inducing inhibition, regression, or stasis of a disease, disorder or condition, or ameliorating or alleviating a symptom of a disease, disorder or condition.
  • “Ameliorating” or “alleviating” a condition or state as used herein shall mean to relieve or lessen the symptoms of that condition or state.
  • “Inhibition” of disease progression or disease complication in a subject as used herein means preventing or reducing the disease progression and/or disease complication in the subject.
  • administering to the subject means the giving of, dispensing of, or application of medicines, drugs, or remedies to a subject to relieve, cure, or reduce the symptoms associated with a condition, e.g., a pathological condition.
  • the drug substance of the present invention may be administered in admixture with suitable pharmaceutical diluents, extenders, excipients, or carriers (collectively referred to herein as a pharmaceutically acceptable carrier) suitably selected with respect to the intended form of administration and as consistent with conventional pharmaceutical practices.
  • a pharmaceutically acceptable carrier suitably selected with respect to the intended form of administration and as consistent with conventional pharmaceutical practices.
  • the unit will be in a form suitable for oral, rectal, topical, intravenous or direct injection or parenteral administration.
  • the compounds can be administered alone or mixed with a pharmaceutically acceptable carrier.
  • This carrier can be a solid or liquid, and the type of carrier is generally chosen based on the type of administration being used.
  • the active agent can be co-administered in the form of a tablet or capsule, liposome, as an agglomerated powder or in a liquid form.
  • suitable solid carriers include lactose, sucrose, gelatin and agar. Capsules or tablets can be easily formulated and can be made easy to swallow or chew; other solid forms include granules, and bulk powders.
  • Capsules or tablets may contain suitable binders, lubricants, disintegrating agents, diluents, coloring agents, flavoring agents, flow-inducing agents, and melting agents.
  • the active drug component can be combined with an oral, non-toxic, pharmaceutically acceptable, inert carrier such as lactose, gelatin, agar, starch, sucrose, glucose, methyl cellulose, magnesium stearate, dicalcium phosphate, calcium sulfate, mannitol, sorbitol and the like.
  • Suitable binders include starch, gelatin, natural sugars such as glucose or beta-lactose, corn sweeteners, natural and synthetic gums such as acacia, tragacanth, or sodium alginate, povidone, carboxymethylcellulose, polyethylene glycol, waxes, and the like.
  • Lubricants used in these dosage forms include sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, sodium chloride, stearic acid, sodium stearyl fumarate, talc and the like.
  • Disintegrators include, without limitation, starch, methyl cellulose, agar, bentonite, xanthan gum, croscarmellose sodium, sodium starch glycolate and the like.
  • the oral dosage form of the present invention may comprise an alkaline-reacting component, said component preferably amounting from about 1 to 20% by weight of the formulation in order to keep the pH above 8.
  • suitable liquid dosage forms include solutions or suspensions in water, pharmaceutically acceptable fats and oils, alcohols or other organic solvents, including esters, emulsions, syrups or elixirs, suspensions, solutions and/or suspensions reconstituted from non-effervescent granules and effervescent preparations reconstituted from effervescent granules.
  • Such liquid dosage forms may contain, for example, suitable solvents, preservatives, emulsifying agents, suspending agents, diluents, sweeteners, thickeners, and melting agents.
  • Oral dosage forms optionally contain flavorants and coloring agents.
  • Parenteral and intravenous forms may also include minerals and other materials to make them compatible with the type of injection or delivery system chosen.
  • the compounds used in the method of the present invention may also be administered in the form of liposome delivery systems, such as small unilamellar vesicles, large unilamallar vesicles, and multilamellar vesicles.
  • Liposomes can be formed from a variety of phospholipids, such as cholesterol, stearylamine, or phosphatidylcholines.
  • the compounds may be administered as components of tissue-targeted emulsions.
  • the compounds used in the method of the present invention may also be coupled to soluble polymers as targetable drug carriers or as a prodrug.
  • soluble polymers include polyvinylpyrrolidone, pyran copolymer, polyhydroxylpropylmethacrylamide-phenol, polyhydroxyethylaspartamidephenol, or polyethyleneoxide-polylysine substituted with palmitoyl residues.
  • the compounds may be coupled to a class of biodegradable polymers useful in achieving controlled release of a drug, for example, polylactic acid, polyglycolic acid, copolymers of polylactic and polyglycolic acid, polyepsilon caprolactone, polyhydroxy butyric acid, polyorthoesters, polyacetals, polydihydropyrans, polycyanoacylates, and crosslinked or amphipathic block copolymers of hydrogels.
  • a class of biodegradable polymers useful in achieving controlled release of a drug
  • a drug for example, polylactic acid, polyglycolic acid, copolymers of polylactic and polyglycolic acid, polyepsilon caprolactone, polyhydroxy butyric acid, polyorthoesters, polyacetals, polydihydropyrans, polycyanoacylates, and crosslinked or amphipathic block copolymers of hydrogels.
  • liquid dosage form For oral administration in liquid dosage form, the oral drug components are combined with any oral, non-toxic, pharmaceutically acceptable inert carrier such as ethanol, glycerol, water, and the like.
  • suitable liquid dosage forms include solutions or suspensions in water, pharmaceutically acceptable fats and oils, alcohols or other organic solvents, including esters, emulsions, syrups or elixirs, suspensions, solutions and/or suspensions reconstituted from non-effervescent granules and effervescent preparations reconstituted from effervescent granules.
  • Such liquid dosage forms may contain, for example, suitable solvents, preservatives, emulsifying agents, suspending agents, diluents, sweeteners, thickeners, and melting agents.
  • Liquid dosage forms for oral administration can contain coloring and flavoring to increase patient acceptance.
  • water a suitable oil, saline, aqueous dextrose (glucose), and related sugar solutions and glycols such as propylene glycol or polyethylene glycols are suitable carriers for parenteral solutions.
  • Solutions for parenteral administration preferably contain a water soluble salt of the active ingredient, suitable stabilizing agents, and if necessary, buffer substances.
  • Antioxidizing agents such as sodium bisulfite, sodium sulfite, or ascorbic acid, either alone or combined, are suitable stabilizing agents.
  • citric acid and its salts and sodium EDTA are also used.
  • parenteral solutions can contain preservatives, such as benzalkonium chloride, methyl- or propyl-paraben, and chlorobutanol.
  • preservatives such as benzalkonium chloride, methyl- or propyl-paraben, and chlorobutanol.
  • Suitable pharmaceutical carriers are described in Remington's Pharmaceutical Sciences, Mack Publishing Company, a standard reference text in this field.
  • the drug substance of the present invention may be administered in various forms, including those detailed herein.
  • the treatment with the compound may be a component of a combination therapy or an adjunct therapy, i.e. the subject or patient in need of the drug is treated or given another drug for the disease in conjunction with one or more of the instant compounds.
  • This combination therapy can be sequential therapy where the patient is treated first with one drug and then the other or the two drugs are given simultaneously. These can be administered independently by the same route or by two or more different routes of administration depending on the dosage forms employed.
  • Gelatin capsules may contain the active ingredient compounds and powdered carriers, such as lactose, starch, cellulose derivatives, magnesium stearate, stearic acid, and the like. Similar diluents can be used to make compressed tablets. Both tablets and capsules can be manufactured as immediate release products or as sustained release products to provide for continuous release of medication over a period of hours. Compressed tablets can be sugar coated or film coated to mask any unpleasant taste and protect the tablet from the atmosphere, or enteric coated for selective disintegration in the gastrointestinal tract.
  • powdered carriers such as lactose, starch, cellulose derivatives, magnesium stearate, stearic acid, and the like. Similar diluents can be used to make compressed tablets. Both tablets and capsules can be manufactured as immediate release products or as sustained release products to provide for continuous release of medication over a period of hours. Compressed tablets can be sugar coated or film coated to mask any unpleasant taste and protect the tablet from the atmosphere, or enteric coated for selective disintegration in the gastrointestinal tract.
  • the compounds used in the method of the present invention may also be administered in intranasal form via use of suitable intranasal vehicles, or via transdermal routes, using those forms of transdermal skin patches well known to those of ordinary skill in that art.
  • the dosage administration will generally be continuous rather than intermittent throughout the dosage regimen.
  • Parenteral and intravenous forms may also include minerals and other materials to make them compatible with the type of injection or delivery system chosen.
  • a dosage unit of the compounds used in the method of the present invention may comprise a single compound or mixtures thereof with additional antibacterial agents.
  • the compounds can be administered in oral dosage forms as tablets, capsules, pills, powders, granules, elixirs, tinctures, suspensions, syrups, and emulsions.
  • the compounds may also be administered in intravenous (bolus or infusion), intraperitoneal, subcutaneous, or intramuscular form, or introduced directly, e.g. by injection, topical application, or other methods, into or onto a site of infection, all using dosage forms well known to those of ordinary skill in the pharmaceutical arts.
  • a “dose” or “dosage unit” of laquinimod as measured in milligrams refers to the milligrams of laquinimod acid present in a preparation, regardless of the form of the preparation.
  • a dosage unit may comprise a single compound or mixtures of compounds thereof.
  • a dosage unit can be prepared for oral dosage forms, such as tablets, capsules, pills, powders, and granules.
  • the “dose” or “dosage unit” of laquinimod may be 0.3, 0.6, or 1.2 mg.
  • a “pharmaceutically acceptable” component is one that is suitable for use with humans and/or animals without undue adverse side effects (such as toxicity, irritation, and allergic response) commensurate with a reasonable benefit/risk ratio.
  • “pharmaceutically acceptable carrier” refers to a carrier or excipient that is suitable for use with humans and/or animals without undue adverse side effects (such as toxicity, irritation, and allergic response) commensurate with a reasonable benefit/risk ratio. It can be a pharmaceutically acceptable solvent, suspending agent or vehicle, for delivering the instant compounds to the subject.
  • the carrier may be liquid or solid and is selected with the planned manner of administration in mind.
  • “Pharmaceutically acceptable carrier” includes “fillers”, which fill out the size of a tablet or capsule, making it practical to produce and convenient for the consumer to use. By increasing the bulk volume, the fillers make it possible for the final product to have the proper volume for patient handling.
  • “Pharmaceutically acceptable carrier” also includes “lubricants”, which prevent ingredients from clumping together and from sticking to the tablet punches or capsule filling machine. Lubricants also ensure that tablet formation and ejection can occur with low friction between the solid and die wall. “Pharmaceutically acceptable carrier” also includes inert carriers such as lactose, gelatin, agar, starch, sucrose, glucose, methyl cellulose, dicalcium phosphate, calcium sulfate, mannitol, sorbitol, microcrystalline cellulose and the like. Liposomes are also a pharmaceutically acceptable carrier.
  • 0.15-0.35% includes 0.15%, 0.16%, 0.17% etc. up to 0.35%.
  • the subject invention is also intended to include all isotopes of atoms occurring on the compounds disclosed herein, including impurities.
  • Isotopes include those atoms having the same atomic number but different mass numbers.
  • isotopes of hydrogen include tritium and deuterium.
  • Isotopes of carbon including C-13 and C-14.
  • detection limit for an analytical method used in screening or testing for the presence of a compound in a sample is a threshold under which the compound in a sample cannot be detected by the analytical method used.
  • the detection limits of a given HPLC method for detecting an impurity in a sample containing laquinimod may vary based on the method and the impurity or impurities being detected.
  • the detection limit of the typical HPLC method for detecting 5-chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid is 0.03% and the detecting limit of a given method for detecting methyl 5-chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylate (MCQME), 5-chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline (MCQ), 5-chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylate (MCQEE) and unknown impurities is 0.02%.
  • MCQME 5-chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylate
  • unknown impurities is 0.02%.
  • quantitation limit for an analytical method used in screening or testing for the presence of a compound in a sample is a threshold under which the compound in a sample cannot be quantified by the analytical method used.
  • the quantitation limits of a given HPLC method for detecting an impurity in a sample containing laquinimod may vary based on the impurity or impurities being detected.
  • density is a measurement defined as the mass of a substance per unit volume.
  • BD bulk density
  • tapped density or “TD” refers to a density measurement of a substance that has been tapped or vibrated, thus minimizing the volume of the substance by eliminating or minimizing the air trapped between particles.
  • rapid stirring refers to stirring which splashes solvent onto the walls of the vessel.
  • Blend uniformity refers to the homogeneity of blend or granulate including laquinimod sodium particles prior to encapsulation, tableting or otherwise finalizing the drug product beyond formation of the final blend, and can represent either one sample or the average of more than one sample.
  • Blend uniformity may be measured, for example, by taking 10 samples that represent the upper, middle and lower layer of each batch of the final blend, performing an HPLC assay to measure the amount of active ingredient in the samples, and comparing the amount of active ingredient in each sample to the labeled amount of active ingredient. The standard deviation and relative standard deviation can be determined based on the individual amounts of the tested samples expressed as percentages of the labeled amount of drug substance in each sample.
  • content uniformity refers to the homogeneity of the laquinimod sodium content among dosage forms, e.g., capsules or tablets, after formulation.
  • Content uniformity may be measured, for example, as indicated by the United States Pharmacopoeia which includes 1) assaying ten tablets (or other dosage form of the drug product) to ensure that the relative standard deviation (RSD) of active content is less than or equal to 6.0% and no value is outside 85-115%; and 2) assaying twenty more tablets (or other dosage form of the drug product) to ensure that the RSD for all thirty is less than or equal to 7.8%, no more than one value is outside 85-115% and no value is outside 75-125% of stated content.
  • RSD relative standard deviation
  • residual solvents include ethanol, n-heptane, n-octane, methanol, acetone, dioxane, and dimethyl formamide. Residual solvents may be determined, for example, based on the manufacturer's statements of residual solvent levels in the active ingredients/excipients and calculation as per U.S. pharmacopeia ⁇ 467> Option 2, product meets the USP ⁇ 467> Residual Solvents limit criteria. Testing is not necessarily required.
  • NMT means no more than.
  • LT means less than.
  • MCQME means methyl 5-chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylate. MCQME is disclosed in U.S. Pat. No. 7,560,557 and J. Org. Chem., 2006, 71, 1658-1667, the contents of which are incorporated by reference into this application. MCQME has the structure:
  • MCQEE means ethyl 5-chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylate. MCQEE is disclosed in U.S. Pat. No. 7,560,557, the contents of which are incorporated by reference into this application. MCQEE has the structure:
  • MCQ means 5-chloro-4-hydroxy-1-methylquinolin-2(1H)-one. MCQ is disclosed in U.S. Pat. No. 7,560,557 and J. Org. Chem., 2006, 71, 1658-1667, the contents of which are incorporated by reference into this application. MCQ has the structure:
  • MCQCA means 5-chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid. MCQCA is disclosed in U.S. Pat. No. 7,560,557 and J. Org. Chem., 2006, 71, 1658-1667, the contents of which are incorporated by reference into this application. MCQCA has the structure:
  • NEA N-ethyl aniline. NEA is disclosed in U.S. Pat. No. 7,560,557, the contents of which are incorporated by reference into this application. NEA has the structure:
  • 5-HLAQ means N-ethyl-4,5-dihydroxy-1-methyl-2-oxo-N-phenyl-1,2-dihydroquinoline-3-carboxamide.
  • 5-HLAQ is disclosed in PCT International Application No. PCT/US13/26476 and U.S. Application Publication No. US 2013/0217724 A1, the contents of which are incorporated by reference into this application.
  • 5-HLAQ has the structure:
  • 3-HLAQ means 5-chloro-N-ethyl-3-hydroxy-1-methyl-2,4-dioxo-N-phenyl-1,2,3,4-tetrahydroquinoline-3-carboxamide.
  • 3-HLAQ is disclosed in PCT International Application No. PCT/US2008/013890 and U.S. Pat. No. 8,178,127 B2, the contents of which are incorporated by reference into this application.
  • 3-HLAQ has the structure:
  • MEG-LAQ means N-ethyl-4-hydroxy-1-methyl-5-(methyl(2,3,4,5,6-pentahydroxyhexyl)amino)-2-oxo-N-phenyl-1,2-dihydroquinoline-3-carboxamide.
  • MEG-LAQ is disclosed in U.S. Application Publication No. US 2013/0345256 A1, the contents of which are incorporated by reference into this application.
  • MEG-LAQ has the structure:
  • DELAQ means 5-chloro-4-hydroxy-1-methyl-2-oxo-N-phenyl-1,2-dihydroquinoline-3-carboxamide.
  • DELAQ is disclosed in PCT International Application No. PCT/US2011/043391 and U.S. Application Publication No. US 2012/0010239 A1, the contents of which are incorporated by reference into this application.
  • DELAQ has the structure:
  • SPIRO-LAQ means 1H,3H-spiro[5-chloro-1-methylquinoline-2,4-dione-3,3′-[l]ethylindolin-[2]-one].
  • SPIRO-LAQ is disclosed in PCT International Application No. PCT/US2008/013890 and U.S. Pat. No. 8,178,127 B2, the contents of which are incorporated by reference into this application.
  • SPIRO-LAQ has the structure:
  • BH-3-HLAQ means 2-chloro-6-(3-(ethyl(phenyl)amino)-2-hydroxy-N-methyl-3-oxopropanamido)benzoic acid.
  • BH-3-HLAQ has the structure:
  • DMM dimethylmalonate
  • DMS Dimethyl sulfate
  • MCIA means 5-chloro-1-methyl-1H-benzo[d][1,3]oxazine-2,4-dione. MCIA has the structure
  • DMF N,N-dimethylformamide
  • LOD loss on drying
  • 0.01 mg to 50 mg means that 0.02, 0.03 . . . 0.09; 0.1, 0.2 . . . 0.9; and 1, 2 . . . 49 mg unit amounts are included as embodiments of this invention.
  • Impurities are measured by common pharmacopeial methods unless otherwise specified.
  • an “anti-solvent” is a solvent in which laquinimod sodium is slightly soluble, very slightly soluble, practically insoluble, or insoluble at room temperature (20-25° C.).
  • solubility terms are defined below, in accordance with the United States Pharmacopoeia XXV.
  • the purification of impure crystalline compounds is usually attained by recrystallization from a suitable solvent or mixture of solvents.
  • the recrystallization process generally comprises the following steps: a) dissolving the impure crystalline substance in a suitable solvent near the boiling point; b) filtering the hot solution from particles of insoluble material and dust; c) allowing the hot solution to cool to cause the dissolved substance to crystallize out; and d) separating the crystals from the supernatant solution.
  • standard recrystallization techniques were accompanied by low or no yields when applied to laquinimod sodium as taught in U.S. Pat. No.
  • the particle size distributions were measured by Malvern Laser Diffraction, using the Mastersizer S model.
  • Laser diffraction relies on the fact that diffraction angle of light is inversely proportional to particle size. Properties of particles are measured and interpreted as measurements of a sphere (a sphere being the only shape that can be described by one unique number).
  • laser diffraction calculates a particle size distribution based around volume terms, thus eliminating particle count from the determination of particle size.
  • the Mastersizer S model measures particles using a single technique and a single range setting.
  • D(0.1) is the particle size, in microns, below which 10% by volume distribution of the population is found.
  • D(0.5) is the particle size, in microns, below which 50% by volume distribution of the population is found.
  • D(0.9) is the particle size, in microns, below which 90% by volume distribution of the population is found.
  • Metal content was measured using inductively coupled plasma atomic emission spectrometry using an inductively coupled plasma atomic emission spectrometry (“ICP-AES”) system manufactured by Spectro (Kleve, Germany). Sample digestion was performed in 65% nitric acid, and the internal standard used was scandium.
  • ICP-AES inductively coupled plasma atomic emission spectrometry
  • Laquinimod sodium and polar impurity/degradation products were determined by isocratic reversed phase high performance liquid chromatography (RP-HPLC), using an ODS-3V column and a mobile phase comprised of a mixture of ammonium acetate buffer at pH 7.0 (80%) and acetonitrile (20%).
  • the detection technique was ultraviolet absorption at 240 nm.
  • Standard stock solution Concentration of standard stock solution is about 300 ⁇ g/mL laquinimod sodium. Standard stock solution may be used for one month when stored in a refrigerator 2° C.-8° C.
  • Laquinimod sodium Concentration of Laquinimod sodium is about 90 ⁇ g/mL. Concentration expressed as laquinimod (acid) is about 85 ⁇ g/mL.
  • Standard working solution A may be used for 7 days when stored in a refrigerator (2° C.-8° C.).
  • Concentration of MCQCA is about 180 ⁇ g/mL.
  • Total dilution factor for laquinimod standard is 1666.67, for MCQCA 2000.
  • Concentration of MCQCA is about 0.09 ⁇ g/mL (0.1%, QL level).
  • Standard solution I may be used for 24 hours when stored in a refrigerator.
  • MCQCA 5-Chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid
  • MCQMA Methyl 5-Chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylate
  • the Mixed Solution may be prepared as follows:
  • Concentration of each impurity in the Mixed Solution is about 30 ⁇ g/mL.
  • Mixed Solution may be used for up to 4 months when stored frozen at about ⁇ 20° C.
  • the freshly prepared Mixed Solution should be divided into aliquots, immediately frozen and stored at ⁇ 20° C. After thawing, the aliquots should be mixed well and should not be refrozen.
  • MCQEE Ethyl 5-Chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylate
  • This solution may be used for up to 4 months when stored frozen at about ⁇ 20° C.
  • the freshly prepared MCQEE Stock Solution should be divided into aliquot, immediately frozen and stored at ⁇ 20° C. After thawing, the aliquots should be mixed well and should not be refrozen.
  • MEG-LAQ Meglumine Adduct of Laquinimod
  • This solution may be used for one week when stored in refrigerator (2° C.-8° C.).
  • Concentration of laquinimod sodium in it is about 90 ⁇ g/mL. Concentration of each impurity—is about 0.9 ⁇ g/mL (about 1% with respect to the working concentration of laquinimod).
  • Resolution Solution 1 is used for resolution test (for system suitability) and for determination of retention times (RT)/relative retention times (RRT) of five impurities: MCQ, MCQCA, MCQME, MCQEE, and 5-HLAQ.
  • Resolution Solution 1 may be used for 9 days if stored in a refrigerator 2° C.-8° C.).
  • Concentration of laquinimod sodium in it is about 90 ⁇ g/mL. Concentration of each impurity—is about 0.9 ⁇ g/mL (about 1% with respect to the working concentration of laquinimod).
  • Resolution Solution 2 is used for determination of retention time of MEG-LAQ.
  • Resolution Solution 2 may be used for 9 days if stored in a refrigerator 2° C.-8° C.).
  • Working concentration of laquinimod (acid) is about 84 ug/mL.
  • sample solutions Immediately after preparation place sample solutions into a refrigerator or in a cooled to 5° C. autosampler. The sample solutions may be used for 24 hours when kept at the temperature 2° C.-8° C.
  • Ignore early elution peaks of excipients and system peaks See chromatogram for determination of impurities/degradation products). For example. Use integration inhibition between 0 and RRT 0.15 (about 2.5 minutes).
  • Typical retention time of laquinimod peak is 15.5 ⁇ 2.0 minutes.
  • Tailing factor (USP) for laquinimod peak should be not more than 2.0.
  • Resolution factor for all the pairs of peaks should be not less than (NLT) 2.
  • MEG-LAQ peak is substantially broadened in comparison with neighboring peaks.
  • Retention time of MEG-LAQ is variable, being very sensitive to slightest changes in chromatographic conditions (pH, % acetonitrile, Temperature, etc.) and therefore should be defined using its peak in the chromatogram of the Resolution Solution 2. Typically, its RRT is about 0.66.
  • MCQCA in Solution I is used to test the sensitivity of the system.
  • RSD of the area of six injections of Std 1 as well as the difference between Std 1 and Std 2 should be no more than (NMT) 20%.
  • Injection diluents B to detect system peaks.
  • the RT of the main peak obtained in the sample chromatogram should correspond to that obtained for the laquinimod peak in the injection of Standard Solution.
  • Area impurity is the area of an impurity/degradation product (known or unknown) peak in the Sample Solution.
  • Area std is the laquinimod peak in chromatogram of Standard Solution I.
  • 0.94 is the conversion factor of laquinimod sodium salt to laquinimod (acid).
  • RRF is the relative response factors of impurities/degradation products calculated as the following ratio: slope of Laquinimod regression line/slope of impurity regression line.
  • MCQME 0.74
  • MCQ 0.65
  • MCQEE 0.85
  • MCQCA 0.62
  • 5-HLAQ 1.0
  • RRF for unknown impurities/degradation products is taken as 1.0.
  • Quantitation level (QL) MCQME, MCQ, MCQEE, 5-HLAQ, and unknown impurities is 005%.
  • Detection level (DL) of MCQME, MCQ, MCQEE, 5-HLAQ and unknown impurities is 0.02%.
  • QL of MCQCA is 0.1%.
  • Detection level DL of MCQCA is 0.03%.
  • Re-crystallization of Laquinimod Na on pilot scale is performed in two glass-lined reactors (Reactor A, 30 liter volume and Reactor B 60 liter). Solid product is filtered and dried in Hastelloy C agitated filter-dryer with 20 micron mesh.
  • Batch size is 2.5 kg of starting crude Laquinimod Na.
  • Reactor A The hot solution in Reactor A is transferred to Reactor B through 0.2 ⁇ m filtration system. Reactor A and filters washed with 1.2 volumes of process water and the wash is transferred to the Reactor B.
  • Vacuum is built-up and the solution in the Reactor B is evaporated at P ⁇ 45 mmHg and jacket temperature T ⁇ 65° C. until volume of the residue reaches 5.4 liters (2.16 volumes). Then atmospheric pressure is built-up and jacket temperature 40-50° C. is adjusted. The batch is stirred for not less than 10 minutes and then seeded with Laquinimod Na crystals to initiate crystallization.
  • the batch is stirred at 45° C. for additional 90 minutes and 7.9 volumes of acetone are added to the reactor in 1.5-2.5 hrs.
  • Reactor temperature during the addition maintained between 40 and 50° C.
  • Resulting slurry is cooled to 0 ⁇ 4° C. during 3.5-4.5 hrs and stirred at this temperature for 10-15 hrs. Then the slurry is transferred to filter-dryer and solid is filtered under pressure of nitrogen.
  • Dry product is discharged, sampled for analysis and packed.
  • Example 15 of U.S. Pat. No. 7,884,208 The pilot scale process of recrystallization of laquinimod sodium was based on Example 15 of U.S. Pat. No. 7,884,208.
  • Example 15 of U.S. Pat. No. 7,884,208 involves 25.0 g of laquinimod sodium (laboratory scale) prepared according to the method disclosed in U.S. Pat. No. 6,875,869.
  • Example 15 the 25.0 g of laquinimod sodium is dissolved in an aqueous solution of laquinimod sodium and then evaporated under vacuum at stirring to a concentrated solution having a volume ratio of 2.14 v/w, the resulting residue is seeded to induce crystallization then treated with an anti-solvent (acetone).
  • an anti-solvent acetone
  • the modified pilot scale process was performed with 2.5 kg of laquinimod sodium which is a 100-fold scale up from Example 15.
  • the modified pilot scale process had significant differences from the laboratory scale process of Example 15 of U.S. Pat. No. 7,884,208. Specifically, evaporation on the laboratory scale was performed in a round-bottom flask in a rotary evaporator without stirring, while evaporation on the pilot scale was performed in a reactor with stirring. On the pilot scale, the evaporation residue is stirred aggressively, liquid splashes on the reactor walls, solid depositions form, and crystallization was spontaneous.
  • the pilot batches did not result in laquinimod sodium particles having a particle size distribution expected based on Example 15 of U.S. Pat. No. 7,884,208. Instead, applicants unexpected found that the pilot batches resulted in a mixture of recrystallized laquinimod sodium particles wherein (i) 90% or more of the total amount by volume of the laquinimod sodium particles have a size of less than 40 microns, (ii) 50% or more of the total amount by volume of the laquinimod sodium particles have a size of less than 15 microns, and (iii) 10% or more of the total amount by volume of the laquinimod sodium particles have a size of less than 5 microns.
  • Table 4 A comparison of particle size distributions resulting from the two different processes is shown in Table 4.
  • laquinimod Na is a potent drug substance, small particle size is advantageous for this API. Formation of non-aggregated laquinimod sodium crystals with reduced particle size could provide better uniformity of drug product and avoid milling or de-lumping operations.
  • the starting material, crude laquinimod sodium appears as aggregated solid.
  • the re-crystallized product is free flowing powder. Powders with smaller particles have a stronger trend to aggregate.
  • Crude laquinimod sodium prepared by slurry-to-slurry recrystallization i.e., the process of U.S. Pat. No. 6,077,851
  • the modified process produces particles having a low particle size and are free flowing.
  • the aim was scalable crystallization procedure giving smaller crystal size, PSD and low aggregation on laboratory, pilot and commercial scale.
  • the desirable PSD profile was the following: d(0.1) ⁇ 5 ⁇ m, d(0.5) ⁇ 15 ⁇ m and d(0.9) ⁇ 40 ⁇ m.
  • the method is based on spontaneous crystallization initiated in aqueous phase prior to acetone addition.
  • the important factor affecting crystallization is initial concentration of crystallizing solution.
  • reduced water volume ratio in the end of evaporation from 2.14 v/w to 1.7-1.8 v/w. Higher concentration of the solution ensures initiation of spontaneous crystallization in the end of evaporation operation and provides higher supersaturation level and lower crystal size.
  • Laquinimod Na re-crystallization step All operations of Laquinimod Na re-crystallization step including evaporation were performed on laboratory scale in transparent agitated glass reactors equipped with stirrer, thermometers and circulating bath for heating and cooling.
  • Vacuum is applied and water is distilled at stirring, pressure during the evaporation is 38-40 mbar and jacket temperature is 55° C.
  • the batch is cooled to 0-5° C. during one hour and filtered on Büchner filter.
  • the solid cake is washed with 75 ml of acetone.
  • Collected wet product (28.0 g) is dried in oven under vacuum at 50° C. to constant weight.
  • Laquinimod Na re-crystallization step All operations of Laquinimod Na re-crystallization step including evaporation were performed on laboratory scale in transparent agitated glass reactors equipped with stirrer, thermometers and circulating bath for heating and cooling.
  • the batch is cooled to 0-5° C. during one hour, stirred at this temperature for one additional hour and filtered on Büchner filter.
  • the solid cake is washed with 75 ml of acetone.
  • Collected wet product (27.5 g) is dried in oven under vacuum at 50° C. to constant weight.
  • Example 2 The results of Example 2 and Example 3 are summarized in Table 5.
  • Table 5 shows that the process reliably produced a mixture of laquinimod sodium crystals a mixture of recrystallized laquinimod sodium particles wherein (i) 90% or more of the total amount by volume of the laquinimod sodium particles have a size of less than 40 microns, (ii) 50% or more of the total amount by volume of the laquinimod sodium particles have a size of less than 15 microns, and (iii) 10% or more of the total amount by volume of the laquinimod sodium particles have a size of less than 5 microns.
  • Recrystallization of laquinimod sodium was performed on a production scale (Batches C, D and E) as follows.
  • Re-crystallization of Laquinimod Na on the production scale is performed in two 250 liter glass-lined reactors (Reactor I and Reactor II). Solid product is filtered and dried in Hastelloy C-22 agitated filter-dryer with 20 micron mesh.
  • Batch size is 6.5-7.5 kg of dry API.
  • Reactor I The hot solution in Reactor I is circulated through 0.2 ⁇ m filtration system at heating and stirring during 15-20 minutes. After the circulation completion filtered solution is transferred to Reactor II through 0.2 ⁇ m filter. Reactor I and filters washed with 1.75 volumes of process water and the wash is transferred to the Reactor II.
  • Vacuum is build-up and the solution in the Reactor II is evaporated at P ⁇ 45 mmHg and jacket temperature T ⁇ 65° C. until volume of the residue reaches 14-16 liter (ca. 1.7-1.8 v/w water/weight crude laquinimod Na starting material). On this step spontaneous crystallization is initiated on the reactor walls. Then atmospheric pressure is build-up and jacket temperature 40-50° C. is adjusted.
  • the batch is stirred for not less than 10 minutes.
  • the batch is stirred at 45° C. for additional 90 minutes and 7.9 volumes of acetone are added to the reactor in 1.5-2.5 hrs.
  • Reactor temperature during the addition maintained between 40 and 50° C.
  • Resulting slurry is cooled to 0 ⁇ 4° C. during 2-5 hrs and stirred at this temperature for 10-15 hrs. Then the slurry is transferred to filter-dryer and solid is filtered under pressure of nitrogen.
  • Dry product is discharged, sampled for analysis and packed.
  • FIGS. 5-8 Microscopic photographs of typical batches of Crude and re-crystallized Laquinimod Na at different magnification are presented on FIGS. 5-8 .
  • the modified crystallization procedure demonstrated good reproducibility of particle size distribution on the production scale. Reduction of the evaporation residue volume to a ratio of 1.7-1.8 v/w and initiation of spontaneous crystallization provided desirable crystal size. Improved crystallization procedure demonstrates reduction in crystal size to a level of d(0.9) ⁇ 40 ⁇ m and good reproducibility of Particle Size Distribution on the production scale.
  • the product with reduced crystal size has no trend to aggregation and does not need milling or de-lumping for homogenization.
  • the product with similar PSD prepared by slurry-to-slurry procedure is aggregated and thus problematic in formulation.
  • the modified crystallization procedure also results in laquinimod sodium having desirable density and purity profiles.
  • the improved crystallization procedure also provides effective purification from organic impurities, e.g., MCQME.
  • Table 9 demonstrates complete removal of MCQME impurity by re-crystallization of commercial scale batch of Laquinimod sodium. Since this intermediate has genotoxic potential it should be purified to undetectable level. The crystallization process also provides purification of all other known organic impurities to the level below the limit of detection.
  • FIGS. 5-8 Microscopic photographs of typical batches of crude and re-crystallized Laquinimod sodium at different magnification are presented on FIGS. 5-8 and show rod-shape morphology of the both products. At the same time, the Crude presented on FIGS. 5 and 6 are much more aggregated than the re-crystallized “Cryst” product of FIGS. 7 and 8 .
  • Laquinimod capsules are manufactured according to the method as described in Example 2 of PCT International Application Publication No. WO 2007/146248, the entire content of which is hereby incorporated by reference. Steps of Example 2 of WO 2007/146248 are performed. Each capsule contains 064 mg of laquinimod sodium equivalent to 0.6 mg laquinimod.
  • the capsules had quantities of impurities within specification based on HPLC relative to the amount of laquinimod.
  • the capsules have a water content of no more than 1.5%.
  • the dissolution profiles, content uniformity, and residual solvents of the encapsulated pharmaceutical composition conforms to U.S. Pharmacopeia ⁇ 711> (dissolution), U.S. Pharmacopeia ⁇ 905> (uniformity), and U.S. Pharmacopeia ⁇ 467>.
  • Each capsule contains 90.0-110.0% of the labeled amount.
  • the capsules contain a total aerobic microbial count (TAMC) of NMT 10 3 cfulg, a total combined yeasts/moulds count (TYMC) of NMT 10 2 cfu/g, and an absence of Escherichia Coli in 1 g.
  • TAMC total aerobic microbial count
  • TYMC total combined yeasts/moulds count
  • Example 5 demonstrates that, in a commercial-scale production, pharmaceutical compositions of laquinimod can be prepared with non-detectable levels or a low level of polar impurities and non-polar impurities.
  • Laquinimod sodium primary reference standard batch was prepared by crystallization of laquinimod sodium batch, followed by purification by crystallization from a water/acetone mixture (1:4.5 w/w). The crystallization was performed by dissolving the drug substance batch in water while heating. The clear solution was filtered and concentrated to a known residue volume under reduced pressure. Acetone was added and the solution was cooled. The obtained crystals were filtered, washed and dried. The chromatographic purity of this batch was found to be 100.0%. The total of both polar and non-polar impurities was LT 0.05%, content of DELAQ was LT 0.1%, and laquinimod acid content was LT 0.2%.
  • MCQ is a white solid.
  • the molecular structure, chemical formula and molecular weight of MCQ are provided below.
  • MCQ was prepared in the following manner: MCQCA and DMSO were heated at 75° C. for 2 hours and were then cooled to room temperature. Water was added and the precipitate was collected by filtration, washed with water and dried in a vacuum oven at 50° C. Material of sufficient purity to be suitable for use as a reference standard was obtained.
  • MCQ was characterized by NMR, MS, elemental analysis and FT-IR as detailed hereafter.
  • the mass spectrum of MCQ was obtained on a Q-TOF Micro-TM-MICROMASS (TOF) mass spectrometer, using electrospray ionization in positive ion mode (ES + ).
  • ES + electrospray ionization in positive ion mode
  • the ratio between MS signals containing 35 Cl and 37 Cl corresponds to the natural abundances of these isotopes (about 3:1).
  • the spectrum shown in FIG. 11 is in agreement with the molecular weight of MCQ.
  • the attribution of the main signals in ES + mass spectrum of MCQ is presented in Table 1.
  • the FT-IR spectrum of MCQ was measured with a Nicolet 6700 “Thermo Scientific” FT-IR apparatus. A representative spectrum is presented in FIG. 12 . A summary of the band assignments is provided in Table 13.
  • MCQCA is a white solid.
  • the molecular structure, chemical formula and molecular weight of MCQCA are provided below.
  • MCQCA was prepared in the following manner:
  • MCQCA was characterized by NMR, MS, elemental analysis and FT-IR as detailed below.
  • the mass spectrum of MCQCA was obtained on a Q-TOF Micro-TM-MICROMASS (TOF) mass spectrometer, using electrospray ionization in positive ion mode (ES + ).
  • ES + electrospray ionization in positive ion mode
  • the ratio between MS signals containing 35 Cl and 37 Cl corresponds to the natural abundances of these isotopes (about 3:1).
  • the spectrum shown in FIG. 15 is in agreement with the molecular weight of MCQCA.
  • the attribution of the main signals in ES + mass spectrum of MCQCA is presented in Table 15.
  • the attenuated total reflectance (ATR) FT-IR spectrum of MCQCA was measured with a Nicolet 6700 “Thermo Scientific” FT-IR apparatus. A representative spectrum is presented in FIG. 16 . A summary of the band assignments is provided in Table 17.
  • MCQME is a white to yellowish solid.
  • the molecular structure, chemical formula and molecular weight of MCQME are provided below.
  • MCQME The preparation of MCQME started with the formation of sodium dimethylmalonate by reaction of dimethylmalonate in DMF with sodium methoxide solution. Then, the intermediate MCIA was reacted with sodium dimethylmalonate to form the MCQME sodium salt. Finally the MCQME sodium salt was acidified to MCQME, which was washed with water and dried. The following is a detailed description of the preparation of MCQME.
  • Aqueous HCl solution (11.1 kg [97.3 mole]) was added during 2 to 3 hours and remains of hydrochloric solution in feeding line were washed out with DMF. The formed slurry is cooled to 20° C. to 30° C. and was maintained at that temperature for not less than 30 minutes.
  • the slurry was transferred gradually to a filter-dryer. Mother liquor was removed by pressing the slurry. Process water (29.8 kg for each cycle) was added to the filter-dryer and pressed out. Washing of the filter-dryer content with water was performed until the pH of outflow washing liquor was NLT 4. The product was dried under vacuum at 40° C. to 50° C. (jacket temperature of filterdryer).
  • Vacuum drying was performed until the LOD of a sample from the filter-dryer was less than 2.0% and the water content (by Karl-Fischer) was not more than 0.5%. Then the jacket was cooled to 20° C. to 30° C. and dry MCQME was discharged.
  • the yield is 62% to 87%.
  • the dry MCQME was delumped, analyzed and released.
  • MCQEE is a light yellow solid.
  • the molecular structure, chemical formula and molecular weight of MCQEE are provided below.
  • MCQEE was prepared in the following manner:
  • MCQEE was characterized by NMR, MS, elemental analysis and FT-IR as detailed below.
  • the mass spectrum of MCQEE was obtained on a Q-TOF Micro-TM-MICROMASS (TOF) mass spectrometer, using electrospray ionization in positive ion mode (ES + ).
  • ES + electrospray ionization in positive ion mode
  • the ratio between MS signals containing 35 Cl and 37 Cl corresponds to the natural abundances of these isotopes (about 3:1).
  • the spectrum shown in FIG. 19 is in agreement with the molecular weight of MCQEE.
  • the attribution of the main signals in ES + mass spectrum of MCQEE is presented in Table 19.
  • the attenuated total reflectance (ATR) FT-IR spectrum of MCQEE was measured with a Nicolet 6700 “Thermo Scientific” FT-IR apparatus, A representative spectrum is presented in FIG. 20 . A summary of the band assignments is provided in Table 21.
  • 5-HLAQ is an off-white solid.
  • the molecular structure, chemical formula and molecular weight of 5-HLAQ are provided below.
  • 5-HLAQ is unstable in most organic solvents as well as in aqueous acidic medium. Therefore purification by the commonly used methods is extremely difficult. Consequently, purification of the material was performed by formation of the di-acetate derivative followed by basic hydrolysis, acidic precipitation and rapid filtration.
  • 5-HLAQ was prepared by a multi-step procedure, depicted below.
  • 5-MeO-laquinimod was obtained by a four-step synthetic procedure starting from 2-amino-6-methoxybenzoic acid (compound 1). These four steps are identical to the manufacturing process of laquinimod starting from ACBA.
  • 5-MeO-laquinimod was dissolved in dichloromethane and the solution was cooled to 0-5° C. AlCl 3 was added and the reaction mixture was stirred for 0.5 hour at 0-5° C. followed by 7 hours' stirring at ambient temperature. The solution was evaporated to dryness at 30° C. (in a water bath) and water was added. The obtained grey solid was filtered, washed with 1N HCl and dried at 30° C. in a vacuum oven.
  • Acetic anhydride was added to a solution of 5-HLAQ crude in pyridine and the reaction mixture was stirred for 1 hour at room temperature.
  • the pyridine was evaporated to dryness and the oily residue was dissolved in dichloromethane.
  • the organic solution was washed with 1N HCl followed by aqueous washings.
  • the crude 2 was purified by flash chromatography on silica gel (mobile phase: 1% methanol in dichloromethane).
  • 5-HLAQ was characterized by NMR, MS, elemental analysis and FT-IR as detailed below.
  • the 1 H-NMR and 13 C-NMR characterization of 5-HLAQ was performed in DMSO on a Bruker 300 MHz instrument.
  • the peak assignments are summarized in Table 22.
  • the 1 H-NMR and 13 C-NMR spectra are presented in FIG. 21 and FIG. 22 , respectively.
  • the mass spectrum of 5-HLAQ was obtained on a Q-TOF Micro-TM-MICROMASS (TOF) mass spectrometer, using electrospray ionization in positive ion mode (ES + ).
  • the attenuated total reflectance (ATR) FT-IR spectrum of 5-HLAQ was measured with a Nicolet 6700 “Thermo Scientific” FT-IR apparatus. A representative spectrum is presented in FIG. 24 . A summary of the band assignments is provided in Table 25.
  • DELAQ is an off-white to pale yellow solid.
  • the molecular structure, chemical formula and molecular weight of DELAQ are provided below.
  • DELAQ was prepared in the following manner:
  • the reaction mixture was cooled to room temperature and n-heptane was added. The crystalline solid formed was collected by filtration and washed with n-heptane. The wet material was dried in a vacuum oven at 50° C.
  • DELAQ was characterized by NMR, MS, elemental analysis and FT-IR, as detailed below.
  • the mass spectrum of DELAQ was obtained on a Q-TOF Micro-TM-MICROMASS (TOF) mass spectrometer, using electrospray ionization in positive ion mode (ES + ).
  • ES + electrospray ionization in positive ion mode
  • the ratio between MS signals containing 35 Cl and 37 Cl corresponds to the natural abundances of these isotopes (about 3:1).
  • the spectrum shown in FIG. 27 is in agreement with the molecular weight of DELAQ.
  • the attribution of the main signals in ES + mass spectrum of DELAQ is presented in Table 27.
  • the FT-IR spectrum of DELAQ was measured with a Nicolet 6700 “Thermo Scientific” FT-IR apparatus. A representative spectrum is presented in FIG. 28 . A summary of the band assignments is provided in Table 29.
  • 3-HLAQ is a yellow solid.
  • the molecular structure, chemical formula and molecular weight of 3-HLAQ are provided below.
  • 3-HLAQ was prepared in the following manner:
  • a solution of laquinimod sodium in water was added drop-wise to a solution of disodium hydrogenphosphate dihydrate and oxone (2KHSO 5 .KHSO 4 .K 2 SO 4 ) in water. 10% NaOH was added until the pH was 8.0. The solution was mixed for 30 minutes at room temperature. The obtained solid was filtered, washed with water and dried in a vacuum oven at 50° C. The crude product was recrystallized twice from ethyl acetate and n-heptane mixture. Each time the product was isolated by cooling the mixture to 10° C. and stirring for 1 hour. The obtained crystals were filtered off, washed with n-heptane and dried in a vacuum oven at 50° C.
  • 3-HLAQ was characterized by NMR, MS, elemental analysis and FT-IR, as detailed below.
  • the mass spectrum of 3-HLAQ was obtained on a Q-TOF Micro-TM-MICROMASS (TOF) mass spectrometer, using electrospray ionization in positive ion mode (ES + ).
  • ES + electrospray ionization in positive ion mode
  • the ratio between MS signals containing 35 Cl and 37 Cl corresponds to the natural abundances of these isotopes (about 3:1).
  • the spectrum shown in FIG. 31 is in agreement with the molecular weight of 3-HLAQ.
  • the attribution of the main signals in ES + mass spectrum of 3-HLAQ is presented in Table 31.
  • the attenuated total reflectance (ATR) FT-IR spectrum of 3-HLAQ was measured with a Nicolet 6700 “Thermo Scientific” FT-IR apparatus. A representative spectrum is presented in FIG. 32 . A summary of the band assignments is provided in Table 33.
  • SPIRO-LAQ is a white powder.
  • the molecular structure, chemical formula and molecular weight of SPIRO-LAQ are provided below.
  • SPIRO-LAQ was prepared in the following manner:
  • SPIRO-LAQ was characterized by NMR, MS, elemental analysis and FT-IR as detailed below.
  • the mass spectrum of SPIRO-LAQ was obtained on a Q-TOF Micro-TM-MICROMASS (TOF) mass spectrometer, using electrospray ionization in positive ion mode (ES + ).
  • ES + electrospray ionization in positive ion mode
  • the ratio between MS signals containing 35 Cl and 37 Cl corresponds to the natural abundances of these isotopes (about 3:1).
  • the spectrum shown in FIG. 35 is in agreement with the molecular weight of SPIRO-LAQ.
  • the attribution of the main signals in ES + mass spectrum of SPIRO-LAQ is presented in Table 35.
  • the attenuated total reflectance (ATR) FT-IR spectrum of SPIRO-LAQ was measured with a Nicolet 6700 “Thermo Scientific” FT-IR apparatus. A representative spectrum is presented in FIG. 36 . A summary of the band assignments is provided in Table 37.
  • MEG-LAQ The molecular structure, chemical formula and molecular weight of MEG-LAQ is presented below.
  • a 1N NaOH solution was added to a suspension of 3-HLAQ in water. The yellow solution was stirred for 0.5 hr followed by the addition of 1N HCl solution. The aqueous solution that contained white solid was extracted with ethyl acetate. The organic phase was washed with brine and dried over sodium sulfate. The suspension was filtered, evaporated to dryness and the solid residue was purified by crystallization from IPA:water mixture (1:3.75 v/v). The suspension was cooled to 0-5° C. and was kept for 1 hr, filtered and washed with IPA:water mixture (1:10 v/v). The obtained white solid was dried in a vacuum oven at 50° C.
  • the BH-3-HLAQ standard was demonstrated to conform to the molecular structure by IR, elemental analysis, MS, and NMR. Additional testing included chromatographic purity and loss on drying.
  • the mass spectrum of BH-3-HLAQ was obtained on a Q-TOF Micro-TM-MICROMASS (TOF) mass spectrometer, using electrospray ionization in positive ion mode (ES + ).
  • ES + electrospray ionization in positive ion mode
  • the spectrum shown in FIG. 39 is in agreement with the molecular weight of BH-3-HLAQ.
  • the attribution of the main signals in ES + mass spectrum of BH-3-HLAQ is presented in Table 39.
  • the attenuated total reflectance (ATR) FT-IR spectrum of BH-3-HLAQ was measured with a Nicolet 6700 “Thermo Scientific” FT-IR apparatus.
  • FIG. 40 shows a typical spectrum. A summary of the band assignments is shown in Table 40.
  • Laquinimod sodium is a white to off-white powder, freely soluble in water which should be stored in a well-closed container, protected from light, at room temperature.
  • Laquinimod sodium The specifications of Laquinimod sodium are as follows:
  • Table 43 lists the non-pharmacopoeial analytical methods currently in use for the determination of impurity levels in the drug substance at the time of release and during stability testing. The method numbers and the detection and quantitation limits for each impurity are also listed.
  • Laquinimod drug product was prepared as 0.6 mg capsules of laquinimod sodium.
  • the specifications of laquinimod sodium (0.6 mg) capsules are as follows:
  • BH-3-HLAQ 2-chloro-6-(3-(ethyl(phenyl)amino)-2-hydroxy-N-methyl-3-oxopropanamido)benzoic acid 7
  • 8 Frequency: For Release: Pilot batches, first 3 production batches and every fifth batch thereafter, or at least one batch per year if less than 5 batches per year are manufactured.
  • Finished product stability protocol package sizes were the largest and smallest of each marketing container/closure and/or unidose configuration.
  • Laquinimod sodium drug substance was analyzed by HPLC.
  • the amount of MCQ, MCQCA, MCQME, MCQEE and 5-HLAQ was determined using the following HPLC method.
  • Buffer pH 7.0 Dissolve 7.7 g of Ammonium acetate in 2000 mL water and adjust to pH 7.0 ⁇ 0.05 with aqueous ammonia or glacial acetic acid. Filter using 0.45 ⁇ m membrane filter. Diluent A Acetonitrile/Water (1:1) Diluent B Mobile phase Blank solution Diluent A/Diluent B (3:7) Note: Adjust flow rate and mobile phase composition to achieve the required system suitability parameters.
  • Standard solution A may be used for 7 days when stored in a refrigerator.
  • Concentration of MCQCA is about 180 ⁇ g/mL.
  • Total dilution factor for Laquinimod standard is 1666.67, for MCQCA 2000.
  • Concentration of MCQCA is about 0.09 ⁇ g/mL (0.1%, QL level).
  • Standard solution I may be used for 24 hours when stored in a refrigerator.
  • each impurity was added to a 100 mL volumetric flask, dissolve and dilute to volume with the Diluent A—Mixed solution. Concentration of each impurity in the Mixed solution is about 30 ⁇ g/mL.
  • the Mixed solution is used only for the determination of the exact retention times of the known impurities and may be used up to four months when stored at about ⁇ 20° C. For this purpose, the freshly prepared Mixed solution is divided into aliquots and stored immediately at about ⁇ 20° C. After thawing, the aliquots should not be refrozen.
  • Concentration of Laquinimod sodium is about 90 ⁇ g/mL.
  • the sample must be analyzed not later than within 24 hours after preparation.
  • the typical retention time for the Laquinimod peak is 15.5 ⁇ 2.0 minutes.
  • the tailing factor for the Laquinimod peak should be not more than 2.0.
  • MCQCA in Solution I is used to test the sensitivity of the system.
  • the RSD of the area of the six injections of Std 1 as well as the difference between Std1 and Std2 should be NMT 20%.
  • the RT of the main (Laquinimod) peak obtained in the sample chromatogram should correspond to that obtained for the Standard solution.
  • Quantitation level of MCQME, MCQ, MCQEE, 5-HLAQ and unknown impurities is 0.05%.
  • Detection level of MCQME, MCQ, MCQEE, 5-HLAQ and unknown impurities is 0.02%.
  • Reporting level of MCQCA is 0.10%.
  • Detection level of MCQCA is 0.03%
  • RRF relative response factors of impurities calculated as the following ratio: slope of Laquinimod sodium regression line/slope of impurity regression line
  • RRF Relative Response Factors
  • Variant 1 At least one of the impurities ⁇ 0.05% or MCQCA ⁇ 0.10%
  • Variant 2 All impurities ⁇ 0.05% and MCQCA ⁇ 0.10%
  • Variant 3 All impurities ⁇ 0.05% and MCQCA ⁇ 0.03%
  • Variant 4 All impurities ⁇ 0.02% and MCQCA ⁇ 0.03%
  • FIG. 41 presents a resolution test chromatogram according to the above method.
  • FIG. 42 presents a diluent chromatogram according to the above method.
  • FIG. 43 presents a typical chromatogram for assay according to the above method.
  • FIG. 44 presents a typical chromatogram for Impurities testing according to the above method.
  • Laquinimod sodium drug substance was analyzed by HPLC.
  • the amount of NEA, 3-HLAQ And SPIRO-LAQ was determined using the following HPLC method.
  • N-Ethylaniline N-Ethylaniline
  • Concentration of stock standard solution is about 500 ⁇ g/mL.
  • NEA stock standard solution may be used for three weeks when stored in a refrigerator.
  • N-Ethylaniline Concentration of N-Ethylaniline is about 5 ⁇ g/mL.
  • N-Ethylaniline Concentration of N-Ethylaniline is about 0.1 g/mL (about 0.1% of the working concentration).
  • NEA working standard solution may be used for four days when stored in a refrigerator.
  • Laquinimod stock solution may be used for one month when stored in refrigerator.
  • Concentration of Laquinimod sodium is about 90 ⁇ g/mL.
  • Concentration of NEA, SPIRO-LAQ and 3-HLAQ is about 1 ⁇ g/mL (1%).
  • the Resolution Solution is used only for the determination of the retention times of Laquinimod, NEA, 3-HLAQ and SPIRO-LAQ and may be used for 38 days when stored at ⁇ 20° C.
  • the freshly prepared resolution solution should be divided into aliquots and stored immediately at ⁇ 20° C. After thawing, the solution aliquots should not be refrozen.
  • the Resolution Solution may be used for 4 days when stored in refrigerator.
  • Concentration of Laquinimod sodium is about 90 ⁇ g/mL.
  • the typical retention time is 2.2 ⁇ 0.5 minutes for Laquinimod peak.
  • the typical retention time is 12.2 ⁇ 1.5 minutes for NEA peak.
  • the chromatographic mapping of the specified impurity peaks should be in the following order: 3-HLAQ, NEA and SPIRO-LAQ.
  • Quantitation limit for NEA and any other impurities is 0.06%
  • Detection limit for NEA and any other impurities is 0.02%.
  • RRF is the Relative Response Factor of NEA to impurities as per the Table:
  • FIG. 45 presents a resolution test chromatogram according to the above method.
  • FIG. 46 presents a diluent chromatogram in according to the above method.
  • FIG. 47 presents a typical sample chromatogram according to the above method.
  • Laquinimod capsules containing 0.6 mg Laquinimod were analyzed by HPLC.
  • the amount of MCQ, MCQCA, 5-HLAQ, MCQME and MCQEE was determined using the following HPLC method.
  • Concentration of standard stock solution is about 300 ⁇ g/mL Laquinimod sodium.
  • Standard stock solution may be used for one month when stored in a refrigerator (2° C.-8° C.).
  • Concentration of Laquinimod sodium is about 90 ⁇ g/mL. Concentration expressed as Laquinimod (acid) is about 85 ⁇ g/mL.
  • Standard working solution A may be used for 7 days when stored in a refrigerator (2° C.-8° C.).
  • Concentration of MCQCA is about 180 ⁇ g/mL.
  • Total dilution factor for Laquinimod standard is 1666.67, for MCQCA 2000.
  • Concentration of MCQCA is about 0.09 ⁇ g/mL (0.1%, QL level).
  • Standard solution I may be used for 24 hours when stored in a refrigerator.
  • the Mixed Solution may be prepared as follows:
  • Concentration of each impurity in the Mixed Solutions is about 30 ⁇ g/mL.
  • Mixed Solution may be used for up to 4 months when stored frozen at about ⁇ 20° C.
  • the freshly prepared Mixed Solution should be divided into aliquots, immediately frozen and stored at ⁇ 20° C. After thawing, the aliquots should be mixed well and should not be refrozen.
  • MCQEE Ethyl 5-chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-carboxylate
  • This Solution may be used for up to 4 months when stored frozen at about ⁇ 20° C.
  • the freshly prepared MCQEE Stock Solution should be divided into aliquots, immediately frozen and stored at ⁇ 20° C. After thawing, the aliquots should be mixed well and should not be refrozen.
  • MEG-LAQ Meglumine Adduct of Laquinimod
  • This Solution may be used for one week when stored in a refrigerator (2° C.-8° C.).
  • Concentration of Laquinimod sodium in it is about 90 ⁇ g/mL. Concentration of each impurity—is about 0.9 g/mL (about 1% with respect to the working concentration of Laquinimod).
  • Resolution Solution 1 is used for resolution test (for system suitability) and for determination of retention times (RT)/relative retention times (RRT) of five known impurities: MCQ, MCQCA, MCQME, MCQEE and 5-HLAQ.
  • Resolution Solution 1 may be used for 9 days if stored in a refrigerator (2° C.-8° C.).
  • Resolution Solution i a Resolution Solution prepared as per the Method of Example 19 may be used.
  • Concentration of Laquinimod sodium in it is about 90 g/mL. Concentration of each impurity—is about 0.9 ⁇ g/mL (about 1% with respect to the working concentration of Laquinimod).

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20200393427A1 (en) * 2018-02-28 2020-12-17 Korea University Research And Business Foundation Metabolome sampling and analysis method for analyzing metabolome during synthetic gas fermentation of synthetic gas fermentation microorganisms

Families Citing this family (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
AU2011223702B2 (en) * 2010-03-03 2016-07-14 Teva Pharmaceutical Industries Ltd. Treatment of lupus arthritis using laquinimod
CN104093310A (zh) 2012-02-03 2014-10-08 泰华制药工业有限公司 拉喹莫德用于治疗一线抗TNFα疗法失败的克罗恩氏病患者的用途
TW201410244A (zh) 2012-08-13 2014-03-16 Teva Pharma 用於治療ｇａｂａ媒介之疾病之拉喹莫德（ｌａｑｕｉｎｉｍｏｄ）
WO2014153145A2 (en) * 2013-03-14 2014-09-25 Teva Pharmaceutical Industries Ltd. Crystals of laquinimod sodium and improved process for the manufacture thereof
MX2016013944A (es) 2014-04-29 2017-01-09 Teva Pharma Laquinimod para el tratamiento de pacientes con esclerosis multiple recidivante-remitente (rrms) con un alto estado de discapacidad.
CN107966506B (zh) * 2017-11-20 2019-09-13 福州大学 一种橡胶及橡胶制品中n-乙基苯胺含量的检测方法

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20070293537A1 (en) * 2006-06-12 2007-12-20 Shulamit Patashnik Stable laquinimod preparations
US7560557B2 (en) * 2002-06-12 2009-07-14 Active Biotech Ag Process for the manufacture of quinoline derivatives
US7884208B2 (en) * 2005-10-19 2011-02-08 Teva Pharmaceutical Industries, Ltd. Crystals of laquinimod sodium, and process for the manufacture thereof
US20120009226A1 (en) * 2008-12-17 2012-01-12 Actavis Group Ptc Ehf Highly pure laquinimod or a pharmaceutically acceptable salt thereof

Family Cites Families (72)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US607785A (en) 1898-07-19 Ore-car
US3024257A (en) 1961-04-10 1962-03-06 Frosst & Co Charles E Stable preparations of alkali metal salts of estrone sulfate
FR2340735A1 (fr) 1976-02-11 1977-09-09 Roussel Uclaf Nouveaux derives de l'acide 3-quinoleine carboxylique, leur procede de preparation et leur application comme medicament
IE52670B1 (en) 1981-03-03 1988-01-20 Leo Ab Heterocyclic carboxamides,compositions containing such compounds,and processes for their preparation
DE3437232A1 (de) 1984-10-10 1986-04-17 Mack Chem Pharm Stabilisierte injektionsloesungen von piroxicam
US4782155A (en) 1986-03-19 1988-11-01 Banyu Pharmaceutical Co., Ltd. Cephalosporin derivatives, processes for their preparation and antibacterial agents
SE8902076D0 (sv) 1989-06-09 1989-06-09 Pharmacia Ab Derivatives of quinoline-3-carboxanilide
HUT60458A (en) 1991-02-01 1992-09-28 Sandoz Ag Process for producing benzyloxyphenyl derivatives and pharmaceutical compositions comprising same
TW376491B (en) 1991-06-22 1999-12-11 Fuji Xerox Co Ltd Image processing system with a buffer memory
US5139878A (en) 1991-08-12 1992-08-18 Allied-Signal Inc. Multilayer film constructions
US5912349A (en) 1997-01-31 1999-06-15 Pharmacia & Upjohn Company Process for the preparation of roquinimex
US20030124187A1 (en) 1997-02-14 2003-07-03 Smithkline Beecham Laboratoires Pharmaceutiques, Pharmaceutical formulations comprising amoxycillin and clavulanate
CA2303867C (en) 1997-09-19 2004-05-11 Cosco, Inc. Playyard
SE9801474D0 (sv) 1998-04-27 1998-04-27 Active Biotech Ab Quinoline Derivatives
US6077851A (en) 1998-04-27 2000-06-20 Active Biotech Ab Quinoline derivatives
US6121287A (en) 1998-07-15 2000-09-19 Active Biotech Ab Quinoline derivatives
US6133285A (en) 1998-07-15 2000-10-17 Active Biotech Ab Quinoline derivatives
SE9802549D0 (sv) 1998-07-15 1998-07-15 Active Biotech Ab Quinoline derivatives
SE9802550D0 (sv) 1998-07-15 1998-07-15 Active Biotech Ab Quinoline derivatives
SE0002320D0 (sv) 1999-10-25 2000-06-21 Active Biotech Ab Malignant tumors
CN100390148C (zh) 1999-10-25 2008-05-28 活跃生物技术有限公司 用于治疗恶性肿瘤的药物
JP2002031610A (ja) 2000-07-14 2002-01-31 Ajinomoto Co Inc 生体分子複合体の界面残基を同定する方法
US6307050B1 (en) 2000-08-29 2001-10-23 R. T. Alamo Venture I Llc Method of synthesizing flosequinan from 4-fluoroanthranilic acid
US6802422B2 (en) 2000-12-12 2004-10-12 Multi-Comp, Inc. Sealed blister assembly
US6706733B2 (en) 2001-05-08 2004-03-16 Dainippon Ink And Chemicals, Inc. Quinolinone derivative formulation and its production method
US20030119826A1 (en) 2001-09-24 2003-06-26 Pharmacia Corporation Neuroprotective treatment methods using selective iNOS inhibitors
US6875869B2 (en) * 2002-06-12 2005-04-05 Active Biotech Ab Process for the manufacture of quinoline derivatives
SE0201778D0 (sv) 2002-06-12 2002-06-12 Active Biotech Ab Process for the manufacture of quinoline derivatives
ITMI20021726A1 (it) 2002-08-01 2004-02-02 Zambon Spa Macrolidi ad attivita' antiinfiammatoria.
US7820145B2 (en) 2003-08-04 2010-10-26 Foamix Ltd. Oleaginous pharmaceutical and cosmetic foam
US7790197B2 (en) 2003-06-09 2010-09-07 Warner-Lambert Company Llc Pharmaceutical compositions of atorvastatin
US20040253305A1 (en) 2003-06-12 2004-12-16 Luner Paul E. Pharmaceutical compositions of atorvastatin
US20050271717A1 (en) 2003-06-12 2005-12-08 Alfred Berchielli Pharmaceutical compositions of atorvastatin
SE0400235D0 (sv) 2004-02-06 2004-02-06 Active Biotech Ab New composition containing quinoline compounds
US8314124B2 (en) 2004-02-06 2012-11-20 Active Biotech Ab Crystalline salts of quinoline compounds and methods for preparing them
KR20130129300A (ko) 2005-02-23 2013-11-27 테바 파마슈티컬 인더스트리즈 리미티드 함량 균일성이 개선된 라사길린 제형
WO2006129178A1 (en) 2005-06-02 2006-12-07 Glenmark Pharmaceuticals S.A. Novel cannabinoid receptor ligands, pharmaceutical compositions containing them, and process for their preparation
AU2007227609A1 (en) 2006-03-16 2007-09-27 Genentech, Inc. Methods of treating lupus using CD4 antibodies
SI2056807T1 (sl) 2006-08-17 2012-12-31 University Of Chicago Zdravljenje vnetnih bolezni
WO2009053070A1 (en) 2007-10-23 2009-04-30 Schwarz Pharma Ag Compounds for treating demyelination conditions
PT2234485E (pt) * 2007-12-20 2014-02-17 Teva Pharma Preparações de laquinimod estáveis
EP2318371A2 (en) 2008-07-01 2011-05-11 Actavis Group PTC EHF Novel solid state forms of laquinimod and its sodium salt
CN102143949A (zh) 2008-09-03 2011-08-03 泰华制药工业有限公司 2-羰基-1，2-二氢喹啉免疫功能调节剂
TW201438738A (zh) 2008-09-16 2014-10-16 Genentech Inc 治療進展型多發性硬化症之方法
UY32427A (es) 2009-02-13 2010-09-30 Boheringer Ingelheim Internat Gmbh Composicion farmaceutica, forma farmaceutica, procedimiento para su preparacion, metodos de tratamiento y usos de la misma
US20100260755A1 (en) 2009-04-09 2010-10-14 Medicinova, Inc. Ibudilast and immunomodulators combination
WO2010147665A1 (en) 2009-06-19 2010-12-23 Teva Pharmaceutical Industries Ltd. Treatment of multiple sclerosis with laquinimod
JP5882208B2 (ja) 2009-07-30 2016-03-09 テバ ファーマシューティカル インダストリーズ リミティド ラキニモドによるクローン病の治療
ES2731052T3 (es) 2009-08-10 2019-11-13 Active Biotech Ab Tratamiento de la enfermedad de Huntington usando laquinimod
AU2011223702B2 (en) 2010-03-03 2016-07-14 Teva Pharmaceutical Industries Ltd. Treatment of lupus arthritis using laquinimod
SG183512A1 (en) 2010-03-03 2012-09-27 Teva Pharma Treatment of lupus nephritis using laquinimod
PH12012501740A1 (en) 2010-03-03 2012-11-12 Teva Pharma Treatment of rheumatoid arthritis with a combination of laquinimod and methotrexate
EP2597954A4 (en) 2010-07-09 2014-01-01 Teva Pharma DEUTERATED N-ETHYL-N-PHENYL-1,2-DIHYDRO-4-HYDROXY-5-CHLORO-1-METHYL-2-OXOCHINOLINE-3-CARBOXAMIDE, SALT THEREOF AND APPLICATIONS THEREOF
NZ606589A (en) 2010-07-09 2015-03-27 Teva Pharma 5-chloro-4-hydroxy-1-methyl-2-oxo-n-phenyl-1,2-dihydroquinoline-3-carboxamide, salts and uses thereof
EP2463289A1 (en) 2010-11-26 2012-06-13 Almirall, S.A. Imidazo[1,2-b]pyridazine derivatives as JAK inhibitors
US20120142730A1 (en) 2010-12-07 2012-06-07 Nora Tarcic Use of laquinimod for reducing fatigue, improving functional status, and improving quality of life in multiple sclerosis patients
KR101130582B1 (ko) 2011-06-07 2012-03-30 이대영 조립식 건축구조물의 골조연결장치
EP2537517A1 (en) 2011-06-22 2012-12-26 Active Biotech AB Deuterium-enriched 4-hydroxy-5-methoxy-n,1-dimethyl-2-oxo-n-[(4-trifluoro-methyl)phenyl]-1,2-dihydroquinoline-3-carboxamide
BR112014002092A2 (pt) 2011-07-28 2017-02-21 Teva Pharma tratamento de esclerose múltipla com combinação de laquinimod e interferon-beta
IN2014MN00333A (enrdf_load_stackoverflow) 2011-07-28 2015-09-25 Teva Pharma
MX2014004420A (es) 2011-10-12 2014-07-09 Teva Pharma Tratamiento de esclerosis multiple con combinacion de laquinimod y fingolimod.
CN104093310A (zh) 2012-02-03 2014-10-08 泰华制药工业有限公司 拉喹莫德用于治疗一线抗TNFα疗法失败的克罗恩氏病患者的用途
EP3225617A1 (en) 2012-02-16 2017-10-04 Teva Pharmaceutical Industries Ltd N-ethyl-n-phenyl-1,2-dihydro-4,5-di-hydroxy-1-methyl-2-oxo-3-quinoline carboxamide, preparation and uses thereof
US20130259856A1 (en) 2012-03-27 2013-10-03 Teva Pharmaceutical Industries, Ltd. Treatment of multiple sclerosis with combination of laquinimod and dimethyl fumarate
TW201804997A (zh) 2012-05-02 2018-02-16 以色列商泰瓦藥品工業有限公司 高劑量拉喹莫德(laquinimod)於治療多發性硬化症之用途
TW201350467A (zh) 2012-05-08 2013-12-16 Teva Pharma Ｎ－乙基－４－羥基－１－甲基－５－（甲基（２，３，４，５，６－五羥基己基）胺基）－２－側氧－ｎ－苯基－１，２－二氫喹啉－３－甲醯胺
WO2014004733A1 (en) 2012-06-26 2014-01-03 The Regents Of The University Of California Composition for lupus nephritis and methods of making and using the same
AR091706A1 (es) 2012-07-11 2015-02-25 Teva Pharma Formulaciones de laquinimod sin agentes alcalinizantes
UY34896A (es) 2012-07-12 2014-02-28 Teva Pharma Tratamiento de la esclerosis múltiple con una combinación de laquinimod y fampridina
TW201410244A (zh) 2012-08-13 2014-03-16 Teva Pharma 用於治療ｇａｂａ媒介之疾病之拉喹莫德（ｌａｑｕｉｎｉｍｏｄ）
TW201410243A (zh) 2012-08-13 2014-03-16 Teva Pharma 用於治療大麻素受體型１（ｃｂ１）媒介之疾病之拉喹莫德（ｌａｑｕｉｎｉｍｏｄ）
WO2014153145A2 (en) * 2013-03-14 2014-09-25 Teva Pharmaceutical Industries Ltd. Crystals of laquinimod sodium and improved process for the manufacture thereof

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7560557B2 (en) * 2002-06-12 2009-07-14 Active Biotech Ag Process for the manufacture of quinoline derivatives
US7884208B2 (en) * 2005-10-19 2011-02-08 Teva Pharmaceutical Industries, Ltd. Crystals of laquinimod sodium, and process for the manufacture thereof
US20070293537A1 (en) * 2006-06-12 2007-12-20 Shulamit Patashnik Stable laquinimod preparations
US20120009226A1 (en) * 2008-12-17 2012-01-12 Actavis Group Ptc Ehf Highly pure laquinimod or a pharmaceutically acceptable salt thereof

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20200393427A1 (en) * 2018-02-28 2020-12-17 Korea University Research And Business Foundation Metabolome sampling and analysis method for analyzing metabolome during synthetic gas fermentation of synthetic gas fermentation microorganisms
US11988651B2 (en) * 2018-02-28 2024-05-21 Korea University Research And Business Foundation Metabolome sampling and analysis method for analyzing metabolome during synthetic gas fermentation of synthetic gas fermentation microorganisms

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