WO2011095802A1 - Procédé hplc pour l'analyse de sunitinib - Google Patents

Procédé hplc pour l'analyse de sunitinib Download PDF

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Publication number
WO2011095802A1
WO2011095802A1 PCT/GB2011/050167 GB2011050167W WO2011095802A1 WO 2011095802 A1 WO2011095802 A1 WO 2011095802A1 GB 2011050167 W GB2011050167 W GB 2011050167W WO 2011095802 A1 WO2011095802 A1 WO 2011095802A1
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WO
WIPO (PCT)
Prior art keywords
salt
sunitinib
hplc method
present
concentration
Prior art date
Application number
PCT/GB2011/050167
Other languages
English (en)
Inventor
Erra Koteswara Satya Vijayakumar
Sachin Bhalekar
Sandesh Parab
Ramdas Bansode
Original Assignee
Generics [Uk] Limited
Mylan India Private Limited
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Filing date
Publication date
Application filed by Generics [Uk] Limited, Mylan India Private Limited filed Critical Generics [Uk] Limited
Publication of WO2011095802A1 publication Critical patent/WO2011095802A1/fr

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Classifications

    • 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
    • G01N30/26—Conditioning of the fluid carrier; Flow patterns
    • G01N30/28—Control of physical parameters of the fluid carrier
    • G01N30/34—Control of physical parameters of the fluid carrier of fluid composition, e.g. gradient
    • 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
    • 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
    • A61K31/40—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with one nitrogen as the only ring hetero atom, e.g. sulpiride, succinimide, tolmetin, buflomedil
    • A61K31/403—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with one nitrogen as the only ring hetero atom, e.g. sulpiride, succinimide, tolmetin, buflomedil condensed with carbocyclic rings, e.g. carbazole
    • A61K31/404—Indoles, e.g. pindolol
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D207/00—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom
    • C07D207/02—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom with only hydrogen or carbon atoms directly attached to the ring nitrogen atom
    • C07D207/30—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having two double bonds between ring members or between ring members and non-ring members
    • C07D207/34—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having two double bonds between ring members or between ring members and non-ring members 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
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D209/00—Heterocyclic compounds containing five-membered rings, condensed with other rings, with one nitrogen atom as the only ring hetero atom
    • C07D209/02—Heterocyclic compounds containing five-membered rings, condensed with other rings, with one nitrogen atom as the only ring hetero atom condensed with one carbocyclic ring
    • C07D209/04—Indoles; Hydrogenated indoles
    • C07D209/30—Indoles; Hydrogenated indoles with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, directly attached to carbon atoms of the hetero ring
    • C07D209/32—Oxygen atoms
    • C07D209/34—Oxygen atoms in position 2
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D403/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00
    • C07D403/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings
    • C07D403/06—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings linked by a carbon chain containing only aliphatic carbon atoms

Definitions

  • the present invention relates to a new HPLC method for the analysis of the drug substance sunitinib and related substances.
  • the present invention relates to a new HPLC method for the analysis of the salt sunitinib malate and related substances.
  • the mobile phase comprises two or more liquids and the relative concentration of the liquids is varied to a predetermined gradient.
  • the mobile phase comprises an alcohol.
  • the mobile phase comprises acetic acid.
  • a fourth method comprises the detection and optional quantification of 5-fluoro-2,3-dihydro-lH- indole-2-one and/ or N-[2-(clietiiylaniino)emyl]-2,4-dimethyl-5-formyl-lH-pyrrole-3- carboxamide.
  • the present invention also relates to sunitinib and associated pharmaceutical compositions from which samples have been analysed by the methods of the invention and/ or which are substantially free of specific impurities.
  • API active pharmaceutical ingredient
  • the likely impurities in APIs and pharmaceutical compositions include residual quantities of synthetic precursors (intermediates), by-products which arise during synthesis of the API, residual solvents, isomers of the API (e.g. geometrical isomers, diastereomers or enantiomers), contaminants which are present in materials used in the synthesis of the API or in the preparation of the pharmaceutical composition, and unidentified adventitious substances.
  • isomers of the API e.g. geometrical isomers, diastereomers or enantiomers
  • contaminants which are present in materials used in the synthesis of the API or in the preparation of the pharmaceutical composition and unidentified adventitious substances.
  • Other impurities which may appear on storage include degradants of the API, for instance formed by hydrolysis or oxidation.
  • the tests that are required to demonstrate that the API or pharmaceutical compositions are safe and effective include purity assay, related substances, content uniformity and dissolution tests.
  • the purity assay test determines the purity of the test product when compared to a standard of a known purity, while the related substances test is used to quantify all the impurities present in the product.
  • the content uniformity test ensures that batches of product like a tablet contain a uniform amount of API and the dissolution test ensures that each batch of product has a consistent dissolution and release of the API.
  • HPLC High Performance Liquid Chromatography
  • the API and the impurities present, if any, are separated on the HPLC stationary phase and they can be detected and quantified using their response obtained from the UV- Visible detector.
  • HPLC is a chromatographic separation technique in which high-pressure pumps force the substance or mixture being analysed together with a mobile phase, also referred to as the eluent, through a separating column containing the stationary phase.
  • HPLC analysis may be performed in isocratic or gradient mode.
  • isocratic mode the mobile phase composition is constant throughout.
  • a gradient HPLC mode is carried out by a gradual change over a period of time in the percentage of the two or more solvents making up the mobile phase. The change in solvent is controlled by a mixer which mixes the solvents to produce the mobile phase prior to its passing through the column. If a substance interacts strongly with the stationary phase, it remains in the column for a relatively long time, whereas a substance that does not interact with the stationary phase as strongly elutes out of the column sooner.
  • the various constituents of the analyte appear at die end of the separating column at different times, known as retention times, where they can be detected and quantified by means of a suitable detector, such as a UV- Visible detector.
  • the current HPLC methods are not particularly suitable for the detection and estimation of total impurities, especially with respect to unknown impurities that are present in a sunitinib or sunitinib malate sample, particularly samples synthesized by the process disclosed in patent US 6573293. Therefore, the HPLC methods reported in the prior art are not particularly convenient or suitable for analysing sunitinib API, particularly with respect to related substances. Consequently, although several HPLC methods have been reported in the prior art for the analysis of sunitinib and its impurities, there is still a need for an alternative method which avoids the problems associated with the known methods as discussed above.
  • An object of the present invention to provide a new, accurate and sensitive HPLC method for the detection and quantitation of all intermediates and related substances that are formed and may remain in the batches of sunitinib whilst avoiding the typical problems associated with the prior art methods.
  • a particular object of the invention is to provide a new, accurate and sensitive HPLC method for the detection and quantitation of all intermediates and related substances that are formed and may remain in the batches of sunitinib or sunitinib malate synthesized by the process disclosed in patent US 6573293. Summary of the invention
  • sunitinib as used herein throughout the description and claims means sunitinib and/ or any salt, solvate, hydrate, anhydrate, tautomer or isomer thereof.
  • the present invention is particularly useful for the analysis of sunitinib (S)-malate.
  • a first aspect of the present invention provides a HPLC method for analysing sunitinib or a salt thereof, wherein the mobile phase comprises two or more liquids and the relative concentration of the liquids is varied to a predetermined gradient.
  • the mobile phase comprises a first liquid A which is aqueous based, such as water or an aqueous solution of a buffer.
  • the buffer is selected from an acid, an organic salt, an inorganic salt, an organic base or a mixture thereof. More preferably the buffer is selected from an acid, an organic salt, an inorganic salt or a mixture thereof.
  • the buffer is a phosphate salt, an acetate salt, a trifluoroacetate salt, a formate salt, acetic acid, trifluoroacetic acid, formic acid, a phosphoric acid such as orthophosphoric acid, or a mixture thereof. More typically, the buffer is a phosphate salt, an acetate salt, a formate salt, acetic acid, trifluoroacetic acid, or a mixture thereof.
  • the buffer is a salt
  • the counter cation is an ammonium cation
  • the buffer is a mixture of an acetate salt, such as ammonium acetate, and an acid, more preferably an organic acid, such as acetic acid.
  • an acetate salt such as ammonium acetate
  • an acid more preferably an organic acid, such as acetic acid.
  • the salt in the buffer can be present at a concentration of 0.001 to 0.2 M, preferably at a concentration of 0.005 to 0.1 M, more preferably at a concentration of 0.01 to 0.1 M, and most preferably at a concentration of about 0.05 M; and/or the acid can be present at a concentration of 0.001 to 0.2 % v/v, preferably at a concentration of 0.005 to 0.1 % v/v, more preferably at a concentration of 0.01 to 0.1 % v/v, and most preferably at a concentration of about 0.05 % v/v.
  • the buffer is a mixture of ammonium acetate present at a concentration of 0.01 to 0.1 M and acetic acid at a concentration of 0.01 to 0.1 % v/v.
  • the buffer is a mixture of ammonium acetate present at a concentration of about 0.05 M and acetic acid at a concentration of about 0.05 % v/v.
  • the alcohol is a C r C 4 alkyl alcohol such as methanol, ethanol, n-propanol, n-butanol, iso-propanol, iso-butanol, sec-butanol or tert-butanol.
  • the second liquid B is methanol.
  • an "alkyl” group is defined as a monovalent saturated hydrocarbon, which may be straight-chained or branched, or be or include cyclic groups.
  • An alkyl group may optionally include one or more heteroatoms N, O or S in its carbon skeleton.
  • Examples of alkyl groups are methyl, ethyl, »-propyl, /-propyl, »-butyl, i- butyl, /-butyl and z-pentyl groups.
  • an alkyl group is straight-chained or branched and does not include any heteroatoms in its carbon skeleton.
  • the second liquid B is substantially water miscible.
  • the magnitude of ⁇ ⁇ is at least 0.10, more preferably at least 0.25, more preferably at least 0.50, more preferably at least 0.75, more preferably at least 0.90, even more preferably at least 0.95.
  • the term "substantially miscible" in relation to two liquids X and Y means that when mixed together at 20°C and 1 atmosphere pressure, X and Y form a single phase when mixed together in any proportion.
  • the mobile phase contains less than 10%, less than 5% or less than 1% acetonitrile by volume. In one embodiment, the mobile phase contains no acetonitrile.
  • the mobile phase contains less than 10%, less than 5% or less than 1% of any organic dipolar aprotic solvent by volume. In one embodiment, the mobile phase contains no organic dipolar aprotic solvent.
  • a preferred embodiment of the first aspect of the present invention is when the first liquid A is an aqueous solution of a buffer comprising an acetate salt mixed with acetic acid, and the second liquid B is methanol.
  • a particularly preferred embodiment of the first aspect of the present invention is when the first liquid A is a mixture of 0.05 M ammonium acetate containing 0.05 % v/v acetic acid and the second liquid B is methanol.
  • the method of the first aspect of the present invention comprises a gradient programming so that the relative concentration of the liquids A and B by volume is typically varied to a gradient between 100 % A : 0 % B to 0 % A : 100 % B over a period of 10 to 180 minutes.
  • the gradient is between 100 % A : 0 % B to 0 % A : 100 % B over a period of 25 to 120 minutes, more preferably 100 % A : 0 % B to 0 % A : 100 % B over a period of 25 to 60 minutes.
  • the HPLC method of the first aspect of the present invention may comprise a gradient programming so that the relative concentration of the liquids A and B by volume starts at a first ratio, then is varied to a first gradient over a first period of time, to arrive at a second ratio, then optionally is maintained at said second ratio for a second period of time.
  • the first ratio may be 70-90 % A : 10-30 % B.
  • the first ratio is 75-85 % A : 15- 25 % B.
  • Most preferably the first ratio is about 80 % A : 20 % B.
  • the first period of time may be from 0 to 120 minutes. Preferably the first period of time is from 15 to 60 minutes. Most preferably the first period of time is about 30 minutes.
  • the second ratio may be 0-20 % A : 80-100 % B. Preferably the second ratio is 5-15 % A : 85-95 % B. Most preferably the second ratio is about 10 % A : 90 % B.
  • the second period of time may be from 0 to 60 minutes. Preferably the second period of time is from 2 to 20 minutes. Most preferably the second period of time is about 5 minutes.
  • the stationary phase used is a gel, preferably a silica gel.
  • the stationary phase used in the first aspect of the present invention is reverse phase such as octadecylsilyl silica gel, octylsilyl silica gel, phenylalkyl silica gel, cyanopropyl silica gel, aminopropyl silica gel or an alkyl-diol silica gel.
  • Particularly suitable stationary phases include octadecylsilyl silica gel or octylsilyl silica gel.
  • a particularly preferred stationary phase comprises an Inertsil ODS 3V (250 mm x 4.6 mm), 5 ⁇ column.
  • the stationary phase has a particle size of between 0.1 and ⁇ , or between 0.5 and 25 ⁇ , or between 1 and ⁇ , or between 4.5 and 6 ⁇ . More preferably the stationary phase has a particle size of about 5 ⁇ .
  • the stationary phase has a pore size of between 10 and ⁇ , or between 25 and 500A, or between 50 and 200A. More preferably the stationary phase has a pore size of between 75 and 125A, or between 90 and llOA. Most preferably the stationary phase has a pore size of about ⁇ .
  • the chromatography is carried out in a column between 10mm and 5000mm in length, or in a column between 50mm and 1000mm in length, or between 100mm and 500mm in length. More preferably the chromatography is carried out in a column between 200mm and 280mm in length. Most preferably the chromatography is carried out in a column about 250mm in length.
  • the chromatography may be carried out in a column between 0.01mm and 100mm in internal diameter, or between O.lmm and 50mm in internal diameter, or between 1mm and 10mm in internal diameter. More preferably the chromatography is carried out in a column about 4.6mm in internal diameter.
  • a particularly preferred method according to the first aspect of the present invention is when the first liquid A is 0.05 M ammonium acetate containing 0.05 % v/v acetic acid and the second liquid B is methanol and the gradient is as follows:
  • the eluent may be analysed by a detector such as a UV and/ or visible spectrophotometer, a fluorescence spectrophotometer, a differential refractometer, an electrochemical detector, a mass spectrometer, a light scattering detector or a radioactivity detector.
  • a detector such as a UV and/ or visible spectrophotometer, a fluorescence spectrophotometer, a differential refractometer, an electrochemical detector, a mass spectrometer, a light scattering detector or a radioactivity detector.
  • the HPLC method detects and optionally quantifies in a single run one or more impurities selected from:
  • the HPLC method detects and optionally quantifies in a single run both 5- fluoro-2,3-dihydro-l H-indole-2-one and N-[2-(diemylammo)ethyl]-2,4-dimethyl-5-formyl- 1 H-pyrrole-3-carboxamide.
  • the HPLC method according to the first aspect of the present invention efficiently detects and quantifies in a single run all impurities including those selected from the following compounds:
  • sunitinib, clozapine and/ or ranitidine is used as internal or external reference marker, or as internal or external reference standard.
  • the sunitinib is in the form of a salt, solvate, hydrate or anhydrate.
  • the method of the first aspect of the present invention is for analysing sunitinib malate. More preferably the method of the first aspect of the present invention is for analysing sunitinib (S)-malate.
  • the method of the first aspect of the present invention is for analysing anhydrous sunitinib, such as anhydrous sunitinib free base or anhydrous sunitinib (S)-malate.
  • the HPLC method is used for the analysis of sunitinib or a salt thereof that is suitable for use in a pharmaceutical composition.
  • the HPLC method is used for the analysis of sunitinib or a salt thereof that has not entered the human or animal body.
  • the sunitinib or the salt thereof that is analysed is not in contact with a human or animal bodily fluid such as plasma.
  • the sunitinib or the salt thereof that is analysed is not in contact with a human or animal bodily tissue such as liver, kidney, brain or white fat tissue.
  • the sunitinib or die salt thereof that is analysed is not in solution.
  • the HPLC mediod is used for the analysis of a pharmaceutical composition comprising sunitinib or a salt thereof.
  • the HPLC method is used for the analysis of a substance comprising at least 5% sunitinib or a salt thereof by weight.
  • the substance comprises at least 10%, at least 25%, at least 50%, at least 75% or at least 90% surtitinib or a salt thereof by weight.
  • the substance comprises at least 95% sunitinib or a salt thereof by weight.
  • the HPLC method is used for the analysis of a substance comprising sunitinib or a salt thereof as the only active pharmaceutical ingredient.
  • a second aspect of the present invention provides a chromatographic method for analysing sunitinib or a salt thereof, wherein the mobile phase comprises an alcohol.
  • the alcohol is a C r C fi alcohol.
  • the alcohol is an alkyl alcohol. More preferably the alcohol is a C r C 4 alkyl alcohol such as methanol, ethanol, n-propanol, n-butanol, iso-ptopanol, iso-butanol, sec-butanol or tert-butanol. Most preferably the alcohol is methanol.
  • the mobile phase comprises tvvo or more liquids, including a first liquid A and a second liquid B, wherein at least one of said liquids comprises the alcohol.
  • the second liquid B is the alcohol.
  • the first liquid A is aqueous based, such as water or an aqueous solution of a buffer.
  • the buffer is selected from an acid, an organic salt, an inorganic salt, an organic base or a mixture thereof. More preferably the buffer is selected from an acid, an organic salt, an inorganic salt or a mixture thereof.
  • the buffer is a phosphate salt, an acetate salt, a trifiuoroacetate salt, a formate salt, acetic acid, ttifluoroacetic acid, formic acid, a phosphoric acid such as orthophosphoric acid, or a mixture thereof. More typically, the buffer is a phosphate salt, an acetate salt, a formate salt, acetic acid, trifluoroacetic acid, or a mixture thereof.
  • the salt in the buffer can be present at a concentration of 0.001 to 0.2 M, preferably at a concentration of 0.005 to 0.1 M, more preferably at a concentration of 0.01 to 0.1 M, and most preferably at a concentration of about 0.05 M; and/ or the acid can be present at a concentration of 0.001 to 0.2 % v/v, preferably at a concentration of 0.005 to 0.1 % v/v, more preferably at a concentration of 0.01 to 0.1 % v/v, and most preferably at a concentration of about 0.05 % v/v.
  • the buffer is a mixture of ammonium acetate present at a concentration of 0.01 to 0.1 M and acetic acid at a concentration of 0.01 to 0.1 % v/v.
  • the buffer is a mixture of ammonium acetate present at a concentration of about 0.05 M and acetic acid at a concentration of about 0.05 % v/v.
  • the pH of the buffer solution is approximately 2 to 6.5. More preferably the pH of the buffer solution is approximately 4 to 6. Most preferably the pH of the buffer solution is about 5.5.
  • the mobile phase contains less than 10%, less than 5% or less than 1% acetonitrile by volume. In one embodiment, the mobile phase contains no acetonitrile. In another embodiment of the second aspect of the present invention, the mobile phase contains less than 10%, less than 5% or less than 1% of any organic dipolar aprotic solvent by volume. In one embodiment, the mobile phase contains no organic dipolar aprotic solvent.
  • a preferred embodiment of the second aspect of the present invention is when the first liquid A is an aqueous solution of a buffer comprising an acetate salt mixed with acetic acid, and the second liquid B is methanol.
  • a particularly preferred embodiment of the second aspect of the present invention is when the first Hquid A is a mixture of 0.05 M ammonium acetate containing 0.05 % v/v acetic acid and the second liquid B is methanol.
  • the chromatographic method is a liquid chromatographic method such as a HPLC, LC-MS or LC-MS/MS method; preferably the chromatographic method is a HPLC method.
  • the chromatographic method may be an isocratic method, preferably such that the relative concentration of the liquids A and B by volume is set between 99.5 % A : 0.5 % B and 0.5 % A : 99.5 % B, or between 90 % A : 10 % B and 10 % A : 90 % B, more preferably between 75 % A : 25 % B and 25 % A : 75 % B. More preferably still the relative concentration of the liquids A and B by volume is about 45 % A : 55 % B. Alternately, the relative concentration of the liquids in the mobile phase may be varied to a predetermined gradient.
  • the relative concentration of the liquids A and B by volume is varied to a gradient between 100 % A : 0 % B to 0 % A : 100 % B over a period of 10 to 180 minutes.
  • the gradient is between 100 % A : 0 % B to 0 % A : 100 % B over a period of 25 to 120 minutes, more preferably 100 % A : 0 % B to 0 % A : 100 % B over a period of 25 to 60 minutes.
  • the chromatographic method of the second aspect of the present invention may comprise a gradient programming so that the relative concentration of the liquids A and B by volume starts at a first ratio, then is varied to a first gradient over a first period of time, to arrive at a second ratio, then optionally is maintained at said second ratio for a second period of time.
  • the first ratio may be 70-90 % A : 10-30 % B. Preferably the first ratio is 75-85 % A : 5- 25 % B. Most preferably the first ratio is about 80 % A : 20 % B.
  • the first period of time may be from 0 to 120 minutes. Preferably the first period of time is from 15 to 60 minutes. Most preferably the first period of time is about 30 minutes.
  • the second ratio may be 0-20 % A : 80-100 % B. Preferably the second ratio is 5-15 % A : 85-95 % B. Most preferably the second ratio is about 10 % A : 90 % B.
  • the second period of time may be from 0 to 60 minutes. Preferably the second period of time is from 2 to 20 minutes. Most preferably the second period of time is about 5 minutes.
  • a particularly preferred method according to the second aspect of the present invention is when the first liquid A is 0.05 M ammonium acetate containing 0.05 % v/v acetic acid and the second liquid B is methanol and the gradient is as follows: Time (min) % A (by volume) % B (by volume)
  • the method of the second aspect of the present invention is carried out at a column temperature between approximately 15 to 40°C.
  • a mobile phase flow rate of between 0.01 and 10 ml/min is used, more preferably a mobile phase flow rate of between 0.1 and 4 ml/min is used, more preferably still a mobile phase flow rate of between 0.5 and 1.5 ml/min is used, most preferably a mobile phase flow rate of about 1 ml/ min is used.
  • the stationary phase used is a gel, preferably a silica gel.
  • the stationary phase used in the second aspect of the present invention is reverse phase such as octadecylsilyl silica gel, octylsilyl silica gel, phenylalkyl silica gel, cyanopropyl silica gel, aminopropyl silica gel or an alkyl-diol silica gel.
  • Particularly suitable stationary phases include octadecylsilyl silica gel or octylsilyl silica gel.
  • a particularly preferred stationary phase comprises an Inertsil ODS 3V (250 mm x 4.6 mm), 5 ⁇ column.
  • the stationary phase has a particle size of between 0.1 and ⁇ , or between 0.5 and 25 ⁇ , or between 1 and ⁇ , or between 4.5 and 6 ⁇ . More preferably the stationary phase has a particle size of about 5 ⁇ .
  • the stationary phase has a pore size of between 10 and ⁇ , or between 25 and 50 ⁇ , or between 50 and 200A. More preferably the stationary phase has a pore size of between 75 and 125A, or between 90 and llOA. Most preferably the stationary phase has a pore size of about lOOA.
  • the chromatography is carried out in a column between 10mm and 5000mm in length, or in a column between 50mm and 1000mm in length, or between 100mm and 500mm in length. More preferably the chromatography is carried out in a column between 200mm and 280mm in length. Most preferably the chromatography is carried out in a column about 250mm in length.
  • the chromatography may be carried out in a column between 0.01mm and 100mm in internal diameter, or between 0.1mm and 50mm in internal diameter, or between 1mm and 10mm in internal diameter. More preferably the chromatography is carried out in a column about 4.6mm in internal diameter.
  • the eluent may be analysed by a detector such as a UV and/ or visible spectrophotometer, a fluorescence spectrophotometer, a differential refractometer, an electrochemical detector, a mass spectrometer, a light scattering detector or a radioactivity detector.
  • a detector such as a UV and/ or visible spectrophotometer, a fluorescence spectrophotometer, a differential refractometer, an electrochemical detector, a mass spectrometer, a light scattering detector or a radioactivity detector.
  • the chromatographic method detects and optionally quantifies in a single run both 5-fluoro-2,3-dihydiO-lH-indole-2-one and N-[2-(dtediylarnino)emyl]-2,4-dimethyl-5- formyl-lH-pyrrole-3-carboxamide.
  • 5-fluoro-2,3-dihydro- lH-indole-2-one and/ or N-[2-(diemylamino)ethyl]-2,4-dimethyl-5-formyl-lH-pyrrole-3- carboxamide is used as internal or external reference marker, or as internal or external reference standard.
  • sunitinib, clozapine and/ or ranitidine is used as internal or external reference marker, or as internal or external reference standard.
  • the sunitinib is in the form of a salt, solvate, hydrate or anhydrate.
  • the method of the second aspect of the present invention is for analysing sunitinib malate. More preferably the method of the second aspect of the present invention is for analysing sunitinib (S)-malate.
  • the method of the second aspect of the present invention is for analysing anhydrous sunitinib, such as anhydrous sunitinib free base or anhydrous sunitinib (S) -malate.
  • the chromatographic method is used for the analysis of sunitinib or a salt thereof that is suitable for use in a pharmaceutical composition.
  • the chromatographic method is used for the analysis of sunitinib or a salt thereof that has not entered the human or animal body.
  • the sunitinib or the salt thereof that is analysed is not in contact with a human or animal bodily fluid such as plasma.
  • the sunitinib or the salt thereof that is analysed is not in contact with a human or animal bodily tissue such as liver, kidney, brain or white fat tissue.
  • the sunitinib or the salt thereof that is analysed is not in solution.
  • the chromatographic method is used for the analysis of a pharmaceutical composition comprising sunitinib or a salt thereof.
  • the chromatographic method is used for the analysis of a substance comprising at least 5% sunitinib or a salt thereof by weight.
  • the substance comprises at least 10%, at least 25%, at least 50%, at least 75% or at least 90% sunitinib or a salt thereof by weight.
  • the substance comprises at least 95% sunitinib or a salt thereof by weight.
  • the chromatographic method is used for the analysis of a substance comprising sunitinib or a salt thereof as the only active pharmaceutical ingredient.
  • a third aspect of the present invention provides a chromatographic method for analysing sunitinib or a salt thereof, wherein the mobile phase comprises acetic acid.
  • the mobile phase comprises an aqueous solution of acetic acid.
  • the acetic acid is typically present at a concentration of 0.001 to 0.2 % v/v, preferably at a concentration of 0.005 to 0.1 % v/v, more preferably at a concentration of 0.01 to 0.1 % v/v, and most preferably at a concentration of about 0.05 % v/v.
  • the aqueous solution further comprises an organic salt, an inorganic salt, an organic base or a mixture thereof. More preferably the aqueous solution further comprises an organic salt, an inorganic salt or a mixture thereof.
  • the salt is a phosphate salt, an acetate salt, a trifiuoroacetate salt, a formate salt, or a mixture thereof.
  • the counter cation to the salt is preferably an ammonium cation.
  • the salt is an acetate salt, such as ammonium acetate.
  • the salt in the aqueous solution can be present at a concentration of 0.001 to 0.2 M, preferably at a concentration of 0.005 to 0.1 M, more preferably at a concentration of 0.01 to 0.1 M, and most preferably at a concentration of about 0.05 M.
  • the mobile phase comprises two or more liquids, including a first liquid A and a second liquid B, wherein at least one of said liquids comprises the acetic acid.
  • the first liquid A is an aqueous solution of the acetic acid. More preferably the first liquid A is an aqueous solution of a mixture of the acetic acid and a salt. Most preferably the first liquid A is an aqueous solution of a mixture of the acetic acid and an acetate salt such as ammonium acetate.
  • the second liquid B preferably comprises or is an organic solvent, preferably selected from an alkyl alcohol, such as methanol, ethanol, propanol or iso-propanol, or acetonitrile or a mixture thereof.
  • an alkyl alcohol such as methanol, ethanol, propanol or iso-propanol, or acetonitrile or a mixture thereof.
  • the second liquid B comprises or is a polar protic organic solvent such as formic acid, methanol, ethanol, n- propanol, n-butanol, iso-propanol, iso-butanol, sec-butanol or tert-butanol, or a mixture thereof.
  • polar protic organic solvent is an alcohol such as a C C 6 alcohol. More preferably the alcohol is an alkyl alcohol.
  • the alcohol is a Q-C 4 alkyl alcohol such as methanol, ethanol, n-propanol, n-butanol, iso-propanol, iso-butanol, sec-butanol or tert-butanol.
  • the second liquid B is methanol.
  • the second liquid B is substantially water miscible.
  • the mobile phase contains less than 10%, less than 5% or less than 1% acetonitrile by volume. In one embodiment, the mobile phase contains no acetonitrile.
  • the mobile phase contains less than 10%, less than 5% or less than 1% of any organic dipolar aprotic solvent by volume. In one embodiment, the mobile phase contains no organic dipolar aprotic solvent.
  • a preferred embodiment of the third aspect of the present invention is when the first liquid A is an aqueous solution of a buffer comprising an acetate salt mixed with acetic acid, and the second liquid B is methanol.
  • a particularly preferred embodiment of the third aspect of the present invention is when the first liquid A is a mixture of 0.05 M ammonium acetate containing 0.05 % v/v acetic acid and the second liquid B is methanol.
  • the chromatographic method is a liquid chromatographic method such as a HPLC, LC-MS or LC-MS/MS method; preferably the chromatographic method is a HPLC method.
  • the relative concentration of the liquids in the mobile phase may be varied to a predetermined gradient.
  • the relative concentration of the liquids A and B by volume is varied to a gradient between 100 % A : 0 % B to 0 % A : 100 % B over a period of 10 to 180 minutes.
  • the gradient is between 100 % A : 0 % B to 0 % A : 100 % B over a period of 25 to 120 minutes, more preferably 100 % A : 0 % B to 0 % A : 100 % B over a period of 25 to 60 minutes.
  • the chromatographic method of the third aspect of the present invention may comprise a gradient programming so that the relative concentration of the liquids A and B by volume starts at a first ratio, then is varied to a first gradient over a first period of time, to arrive at a second ratio, then optionally is maintained at said second ratio for a second period of time.
  • the first ratio may be 70-90 % A : 10-30 % B. Preferably the first ratio is 75-85 % A : 15 25 % B. Most preferably the first ratio is about 80 % A : 20 % B.
  • the first period of time may be from 0 to 120 minutes. Preferably the first period of time is from 15 to 60 minutes. Most preferably the first period of time is about 30 minutes.
  • the second ratio may be 0-20 % A : 80-100 % B. Preferably the second ratio is 5-15 % A 85-95 % B. Most preferably the second ratio is about 10 % A : 90 % B.
  • the second period of time may be from 0 to 60 minutes. Preferably the second period of time is from 2 to 20 minutes. Most preferably the second period of time is about 5 minutes.
  • a particularly preferred method according to the third aspect of the present invention is when the first liquid A is 0.05 M ammonium acetate containing 0.05 % v/v acetic acid and the second liquid B is methanol and the gradient is as follows:
  • the method of the third aspect of the present invention is carried out at a column temperature between approximately 15 to 40°C.
  • a mobile phase flow rate of between 0.01 and 10 ml/min is used, more preferably a mobile phase flow rate of between 0.1 and 4 ml/min is used, more preferably still a mobile phase flow rate of between 0.5 and 1.5 rm/min is used, most preferably a mobile phase flow rate of about 1 ml/ min is used.
  • the stationary phase has a particle size of between 0.1 and ⁇ , or between 0.5 and 25 ⁇ , or between 1 and ⁇ , or between 4.5 and 6 ⁇ . More preferably the stationary phase has a particle size of about 5 ⁇ .
  • the stationary phase has a pore size of between 10 and ⁇ , or between 25 and 500A, or between 50 and 20 ⁇ . More preferably the stationary phase has a pore size of between 75 and 125A, or between 90 and HOA. Most preferably the stationary phase has a pore size of about ⁇ .
  • the chromatography is carried out in a column between 10mm and 5000mm in length, or in a column between 50mm and 1000mm in length, or between 100mm and 500mm in length. More preferably the chromatography is carried out in a column between 200mm and 280mm in length. Most preferably the chromatography is carried out in a column about 250mm in length.
  • the chromatography may be carried out in a column between 0.01mm and 100mm in internal diameter, or between 0.1mm and 50mm in internal diameter, or between 1mm and 10mm in internal diameter. More preferably the chromatography is carried out in a column about 4.6mm in internal diameter.
  • the eluent may be analysed by a detector such as a UV and/ or visible spectrophotometer, a fluorescence spectrophotometer, a differential refractometer, an electrochemical detector, a mass spectrometer, a light scattering detector or a radioactivity detector.
  • a detector such as a UV and/ or visible spectrophotometer, a fluorescence spectrophotometer, a differential refractometer, an electrochemical detector, a mass spectrometer, a light scattering detector or a radioactivity detector.
  • the chromatographic method detects and optionally quantifies in a single run one or more impurities selected from: 5-fluoro-2,3-d ydro-lH-indole-2-one; and
  • the chromatographic method detects and optionally quantifies in a single run both 5-fluoro-2,3-dihydro-lH-indole-2-one and N-[2-(cHethylammo)ethyl]-2,4-dimethyl-5- formyl-lH-pyrrole-3-carboxamide.
  • sunitinib, clozapine and/ or ranitidine is used as internal or external reference marker, or as internal or external reference standard.
  • the sunitinib is in the form of a salt, solvate, hydrate or anhydrate.
  • the method of the third aspect of the present invention is for analysing sunitinib malate. More preferably the method of the third aspect of tlie present invention is for analysing sunitinib (S)-malate.
  • the method of the third aspect of the present invention is for analysing anhydrous sunitinib, such as anhydrous sunitinib free base or anhydrous sunitinib (S)-malate.
  • the chromatographic method is used for the analysis of sunitinib or a salt thereof that is suitable for use in a pharmaceutical composition.
  • the chromatographic method is used for the analysis of sunitinib or a salt thereof that has not entered the human or animal body.
  • the sunitinib or the salt thereof that is analysed is not in contact with a human or animal bodily fluid such as plasma.
  • the sunitinib or the salt thereof that is analysed is not in contact with a human or animal bodily tissue such as liver, kidney, brain or white fat tissue.
  • the sunitinib or the salt thereof that is analysed is not in solution.
  • the chromatographic method is used for the analysis of a pharmaceutical composition comprising sunitinib or a salt thereof.
  • the chromatographic method is used for the analysis of a substance comprising at least 5% sunitinib or a salt thereof by weight.
  • the substance comprises at least 10%, at least 25%, at least 50%, at least 75% or at least 90% sunitinib or a salt thereof by weight.
  • the substance comprises at least 95% sunitinib or a salt thereof by weight.
  • the chromatographic method is used for the analysis of a substance comprising sunitinib or a salt thereof as the only active pharmaceutical ingredient.
  • a fourth aspect of the present invention provides a method for analysing a substance, comprising the detection and optional quantification of 5-fluoro-2,3-dihydro-lH-indole-2- one and/ or N-[2-( ⁇ diethylamino)ethyl]-2,4-dim
  • the method of the fourth aspect of the present invention comprises the detection and optional quantification of both 5-fiuoro-2,3-dihydro-lH-indole-2-one and N- [2-((iiethylamino)ethyl]-2,4-dimethyl-5-formyl-lH-pyrrole-3-carboxami
  • the method further comprises the detection and optional quantification of sunitinib or a salt thereof.
  • the substance is an active pharmaceutical ingredient.
  • the substance is sunitinib, optionally in the form of a salt, solvate, hydrate or anhydrate.
  • the sunitinib is in the form of the malate salt. Most preferably the sunitinib is in the form of the (S)-malate salt.
  • the sunitinib is anhydrous, such as anhydrous sunitinib free base or anhydrous sunitinib (S)-malate.
  • the sunitinib or the salt thereof analysed is for use in a pharmaceutical composition.
  • the substance that is analysed has not entered the human or animal body.
  • the substance that is analysed is not in contact with a human or animal bodily fluid such as plasma.
  • the substance that is analysed is not in contact with a human or animal bodily tissue such as liver, kidney, brain or white fat tissue.
  • the substance that is analysed is not in solution.
  • the method is a method of analysing a pharmaceutical composition comprising sunitinib or a salt thereof.
  • the substance comprises at least 5% sunitinib or a salt thereof by weight.
  • the substance comprises at least 10%, at least 25%, at least 50%, at least 75% or at least 90% sunitinib or a salt thereof by weight.
  • the substance comprises at least 95% sunitinib or a salt thereof by weight.
  • the substance comprises sunitinib or a salt thereof as the only active pharmaceutical ingredient.
  • the first liquid A is aqueous based, such as water or an aqueous solution of a buffer.
  • the buffer is selected from an acid, an organic salt, an inorganic salt, an organic base or a mixture thereof. More preferably the buffer is selected from an acid, an organic salt, an inorganic salt or a mixture thereof.
  • the buffer is a phosphate salt, an acetate salt, a trifluoroacetate salt, a formate salt, acetic acid, trifluoroacetic acid, formic acid, a phosphoric acid such as orthophosphoric acid, or a mixture thereof. More typically, the buffer is a phosphate salt, an acetate salt, a formate salt, acetic acid, trifluoroacetic acid, or a mixture thereof.
  • the buffer is a salt
  • the counter cation is an ammonium cation
  • the salt in the buffer can be present at a concentration of 0.001 to 0.2 M, preferably at a concentration of 0.005 to 0.1 M, more preferably at a concentration of 0.01 to 0.1 M, and most preferably at a concentration of about 0.05 M; and/or the acid can be present at a concentration of 0.001 to 0.2 % v/v, preferably at a concentration of 0.005 to 0.1 % v/v, more preferably at a concentration of 0.01 to 0.1 % v/v, and most preferably at a concentration of about 0.05 % v/v.
  • the second liquid B comprises or is a polar protic organic solvent such as acetic acid, methanol, ethanol, n- propanol, n-butanol, iso-propanol, iso-butanol, sec-butanol or tert-butanol, or a mixture thereof.
  • polar protic organic solvent is an alcohol such as a Q-C 6 alcohol. More preferably the alcohol is an alkyl alcohol.
  • the alcohol is a C r C 4 alkyl alcohol such as methanol, ethanol, n-propanol, n-butanol, iso-propanol, iso-butanol, sec-butanol or tert-butanol.
  • the second liquid B is methanol.
  • the second liquid B is substantially water miscible.
  • the mobile phase contains less than 10%, less than 5% or less than 1% acetonitrile by volume. In one embodiment, the mobile phase contains no acetonitrile.
  • a preferred embodiment of the fourth aspect of the present invention is when the first liquid A is an aqueous solution of a buffer comprising an acetate salt mixed with acetic acid, and the second liquid B is methanol.
  • a particularly preferred embodiment of the fourth aspect of the present invention is when the first liquid A is a mixture of 0.05 M ammonium acetate containing 0.05 % v/v acetic acid and the second liquid B is methanol.
  • the chromatographic method is a liquid chromatographic method such as a HPLC, LC-MS or LC-MS/MS method; preferably the chromatographic method is a HPLC method.
  • said method detects and optionally quantifies in a single run 5-fluoro-2,3-dihydro-lH- indole-2-one and/ or N-[2-(diemylamino)emyl]-2,4-dimethyl-5-formyl-lH-pyrrole-3- carboxamide. More preferably said method also detects and optionally quantifies in the same run sunitinib or a salt thereof.
  • said method detects and optionally quantifies in a single run all three of: sunitinib or a salt thereof;
  • 5-fluoro-2,3-dihydro- lH-indole-2-one and/ or N-[2-(diemylaniino)emyl]-2,4-dimethyl-5-formyl-lH-pyrrole-3- carboxamide is used as internal or external reference marker, or as internal or external reference standard.
  • sunitinib, clozapine and/ or ranitidine is used as internal or external reference marker, or as internal or external reference standard.
  • the chromatographic method may be an isocratic method, preferably such that the relative concentration of the liquids A and B by volume is set between 99.5 % A : 0.5 % B and 0.5 % A : 99.5 % B, or between 90 % A ; 10 % B and 10 % A : 90 % B, more preferably between 75 % A : 25 % B and 25 % A : 75 % B. More preferably still the relative concentration of the liquids A and B by volume is about 45 % A : 55 % B.
  • the relative concentration of the liquids in the mobile phase may be varied to a predetermined gradient.
  • the relative concentration of the liquids A and B by volume is varied to a gradient between 100 % A : 0 % B to 0 % A : 100 % B over a period of 10 to 180 minutes.
  • the gradient is between 100 % A : 0 % B to 0 % A : 100 % B over a period of 25 to 120 minutes, more preferably 100 % A : 0 % B to 0 % A : 100 % B over a period of 25 to 60 minutes.
  • the chromatographic method of the fourth aspect of the present invention may comprise a gradient programming so that the relative concentration of the liquids A and B by volume starts at a first ratio, then is varied to a first gradient over a first period of time, to arrive at a second ratio, then optionally is maintained at said second ratio for a second period of time.
  • the first ratio may be 70-90 % A : 10-30 % B. Preferably the first ratio is 75-85 % A : 15- 25 % B. Most preferably the first ratio is about 80 % A : 20 % B.
  • the first period of time may be from 0 to 120 minutes. Preferably the first period of time is from 15 to 60 minutes. Most preferably the first period of time is about 30 minutes.
  • the second ratio may be 0-20 % A : 80-100 % B. Preferably the second ratio is 5-15 % A : 85-95 % B. Most preferably the second ratio is about 10 % A : 90 % B.
  • the second period of time may be from 0 to 60 minutes. Preferably the second period of time is from 2 to 20 minutes. Most preferably the second period of time is about 5 minutes.
  • the stationary phase used is a gel, preferably a silica gel.
  • the stationary phase used in the fourth aspect of the present invention is reverse phase such as octadecylsilyl silica gel, octylsilyl silica gel, phenylalkyl silica gel, cyanopropyl silica gel, aminopropyl silica gel or an alkyl-diol silica gel.
  • Particularly suitable stationary phases include octadecylsilyl silica gel or octylsilyl silica gel.
  • a particularly preferred stationary phase comprises an Inertsil ODS 3V (250 mm x 4.6 mm), 5 ⁇ column.
  • the stationary phase has a particle size of between 0.1 and ⁇ , or between 0.5 and 25 ⁇ , or between 1 and ⁇ , or between 4.5 and 6 ⁇ . More preferably the stationary phase has a particle size of about 5 ⁇ .
  • the stationary phase has a pore size of between 10 and ⁇ , or between 25 and 500A, or between 50 and 200A. More preferably the stationary phase has a pore size of between 75 and 125A, or between 90 and HOA. Most preferably the stationary phase has a pore size of about ⁇ .
  • the chromatography is carried out in a column between 10mm and 5000mm in length, or in a column between 50mm and 1000mm in length, or between 100mm and 500mm in length. More preferably the chromatography is carried out in a column between 200mm and 280mm in length. Most preferably the chromatography is carried out in a column about 250mm in length.
  • the chromatography may be carried out in a column between 0.01mm and 100mm in internal diameter, or between O.lmm and 50mm in internal diameter, or between 1mm and 10mm in internal diameter. More preferably the chromatography is carried out in a column about 4.6mm in internal diameter.
  • the eluent may be analysed by a detector such as a UV and/ or visible spectrophotometer, a fluorescence spectrophotometer, a differential refractometer, an electrochemical detector, a mass spectrometer, a light scattering detector or a radioactivity detector.
  • a detector such as a UV and/ or visible spectrophotometer, a fluorescence spectrophotometer, a differential refractometer, an electrochemical detector, a mass spectrometer, a light scattering detector or a radioactivity detector.
  • a fifth aspect of the present invention provides a process for preparing a batch of a substance, said process comprising the steps of:
  • the substance comprises or is an active pharmaceutical ingredient.
  • the substance comprises or is sunitinib, optionally in the form of a salt, solvate, hydrate or anhydrate. More preferably the sunitinib is in the form of the malate salt. Most preferably the sunitinib is in the form of the (S)- malate salt.
  • the sunitinib is anhydrous, such as anhydrous sunitinib free base or anhydrous sunitinib (S)-malate.
  • the substance is for use in a pharmaceutical composition.
  • the substance comprises or is a pharmaceutical composition.
  • the pharmaceutical composition comprises sunitinib, optionally in the form of a salt, solvate, hydrate or anhydrate. More preferably the sunitinib is in the form of the malate salt. Most preferably the sunitinib is in the form of the (S)-makte salt.
  • the sunitinib is anhydrous, such as anhydrous sunitinib free base or anhydrous sunitinib (S)-malate.
  • the pharmaceutical composition comprises one or more pharmaceutically acceptable excipients.
  • the substance of the fifth aspect of the present invention has not entered the human or animal body.
  • the substance is not in contact with a human or animal bodily fluid such as plasma.
  • the substance is not in contact with a human or animal bodily tissue such as liver, kidney, brain or white fat tissue.
  • the substance is not in solution.
  • the substance comprises at least 5% sunitinib or a salt thereof by weight.
  • the substance comprises at least 10%, at least 25%, at least 50%, at least 75% or at least 90% sunitinib or a salt thereof by weight.
  • the substance comprises at least 95% sunitinib or a salt thereof by weight.
  • the substance comprises sunitinib or a salt thereof as the only active pharmaceutical ingredient.
  • a sixth aspect of the present invention provides a batch of sunitinib or a salt thereof which has been prepared by a process according to the fifth aspect of the present invention.
  • the sunitinib is substantially free of 5-fluoro-2,3-dihydro-lH-indole-2-one and /or N- [2- (dietiiylamino) ethyl] -2,4-dimethyl-5-formyl- 1 H-pyrrole-3-carboxamide.
  • Sumtinib or a salt thereof is "substantially free" of a compound, if it comprises less than about 5% of that compound, preferably less than about 3%, preferably less than about 2%, preferably less than about 1%, preferably less than about 0.5%, preferably less than about 0.1%, preferably less than about 0.05%, preferably as measured by HPLC.
  • a seventh aspect of the present invention provides a process for preparing a pharmaceutical composition, said process comprising the step of combining one or more pharmaceutically acceptable excipients with part or all of a batch of sunitinib or a salt thereof which has been prepared by a process according to the fifth aspect of the present invention.
  • An eighth aspect of the present invention provides a pharmaceutical composition prepared by a process according to the seventh aspect of the present invention.
  • a ninth aspect of the present invention provides a batch of one or more pharmaceutical compositions which have been prepared by a process according to the fifth aspect of the present invention, wherein the pharmaceutical composition(s) comprise sunitinib or a salt thereof.
  • the pharmaceutical composition(s) also comprise one or more pharmaceutically acceptable excipients.
  • any embodiment of a given aspect of the present invention may occur in combination with any other embodiment of the same aspect of the present invention.
  • any preferred or optional embodiment of any aspect of the present invention should also be considered as a preferred or optional embodiment of any other aspect of the present invention.
  • the present invention can be used to analyse sunitinib and/ or its salts, in particular sunitinib malate, as an API or when prepared as a pharmaceutical composition.
  • compositions that can be analysed by the present invention include solid and liquid compositions and optionally comprise one or more pharmaceutically acceptable carriers or excipients.
  • Solid form compositions include powders, tablets, pills, capsules, cachets, suppositories, and dispersible granules.
  • Liquid compositions include solutions or suspensions which can be administered by oral, injectable, inhalation or infusion routes.
  • impurities of "related substances” as used herein throughout the specification can mean either impurities formed in the manufacture of the API or the pharmaceutical composition and/or formed by degradation of the API or in the pharmaceutical composition on storage.
  • the present invention is also advantageous as the method is selective, linear and precise for the analysis of related substances in sunitinib and/or its salts.
  • the present invention is highly sensitive and allows detection and quantification of related substances in sunitinib and/ or its salts at levels much lower than acceptance limits specified by health authorities.
  • the method of the present invention can be used to easily detect and quantify all degradation impurities formed on storage of samples of sunitinib. This was established by carrying out forced degradation studies as per ICH Q1A (R2) Guidelines and validated as per ICH Q2C (Rl) Guidelines covering the parameters Specificity, Linearity and Range, Precision (Reproducibility), Limit of Detection (LOD), Limit of Quantitation (LOQ) and System Suitability.
  • the buffer optionally used in the first liquid A can be an inorganic salt such as sodium, potassium, calcium, magnesium, lithium or aluminium salts of phosphate, acetate or formate and mixtures thereof.
  • the buffer can be an organic salt such as the ammonium salt of acetate or formate and mixtures thereof.
  • the buffer can be a mineral acid or a carboxylic acid, such as acetic acid or trifluoroacetic acid.
  • the first liquid A is a mixture of 0.05 M ammonium acetate and 0.05 % v/v acetic acid.
  • the second liquid B is an organic solvent such as an alcohol, preferably a C, to C 6 alkyl alcohol like methanol, ethanol, propanol, butanol or iso-propanol or mixtures thereof.
  • the organic solvent(s) may be tetrahydrofuran, ethyl acetate or acetonitrile or any suitable organic solvent(s).
  • the organic solvent is methanol.
  • the stationary phase used in the method of the present invention is selected from octadecylsilyl silica gel (RP-18) or octylsilyl silica gel (RP-8).
  • An internal standard reference compound may be used in the method of the present invention if required.
  • concentration of the components analysed may be determined by comparison with one or more external reference compounds.
  • the inventors have tested the methods of the present invention extensively to show that they are reproducible, precise and linear with respect to concentration.
  • the sample of sunitinib (S)-malate (anhydrous) is initially dissolved in a small volume of the diluent; the sample solution is then injected into the column which is run using the mobile phase outlined below;
  • First Liquid A a mixture of 0.05 M ammonium acetate and 0.05 % v/v acetic acid;
  • RT Retention times
  • RRT Relative retention times
  • LOD Limit of Detection
  • LOQ Limit of Quantitation

Abstract

La présente invention concerne un nouveau procédé HPLC pour l'analyse de la substance médicamenteuse sunitinib et de substances apparentées. La présente invention concerne notamment un nouveau procédé HPLC pour l'analyse du sel malate de sunitinib et de substances apparentées. Dans un premier procédé, la phase mobile comprend deux liquides ou plus et la concentration relative des liquides est variée selon un gradient prédéterminé. Dans un deuxième procédé, la phase mobile comprend un alcool. Dans un troisième procédé, la phase mobile comprend de l'acide acétique. Un quatrième procédé comprend la détection et éventuellement la quantification de 5-fluoro-2,3-dihydro-1H-indole-2-one et/ou de N-[2-(diéthylamino)éthyl]-2,4-diméthyl-5-formyl-1H-pyriole-3-carboxamide. La présente invention concerne également le sunitinib et des compositions pharmaceutiques associées à partir desquelles des échantillons ont été analysés par les procédés de l'invention et/ou qui sont essentiellement exemptes d'impuretés spécifiques.
PCT/GB2011/050167 2010-02-02 2011-02-01 Procédé hplc pour l'analyse de sunitinib WO2011095802A1 (fr)

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WO2017148418A1 (fr) * 2016-03-03 2017-09-08 石家庄以岭药业股份有限公司 Procédé de détermination du contenu des composants d'une composition de médecine chinoise
CN112098536A (zh) * 2020-08-19 2020-12-18 四川省肿瘤医院 一种测定人血浆中舒尼替尼浓度的方法
CN112684057A (zh) * 2020-12-31 2021-04-20 南京品生医疗科技有限公司 一种检测血清中11种抗肿瘤药物浓度的试剂盒及其应用
CN115060819A (zh) * 2022-06-08 2022-09-16 重庆大学附属肿瘤医院 基于hplc-ms/ms单峰法同时测定人血浆中sun及su12662的方法
CN115060819B (zh) * 2022-06-08 2023-09-01 重庆大学附属肿瘤医院 基于hplc-ms/ms单峰法同时测定人血浆中sun及su12662的方法

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