WO2017018820A1 - Pharmaceutical composition with improved storage stability and method for preparing the same - Google Patents

Pharmaceutical composition with improved storage stability and method for preparing the same Download PDF

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Publication number
WO2017018820A1
WO2017018820A1 PCT/KR2016/008270 KR2016008270W WO2017018820A1 WO 2017018820 A1 WO2017018820 A1 WO 2017018820A1 KR 2016008270 W KR2016008270 W KR 2016008270W WO 2017018820 A1 WO2017018820 A1 WO 2017018820A1
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pharmaceutical composition
weight
composition according
amount
formula
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PCT/KR2016/008270
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French (fr)
Inventor
Bong Oh Kim
Kyu Jin Kyung
Ji Yeong Kim
Hye Rim Kim
Bum Chan MIN
Yoo Jeong YOON
Min Hyo Seo
Yil Woong Yi
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Samyang Biopharmaceuticals Corporation
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Priority to RU2018107055A priority Critical patent/RU2018107055A/en
Priority to CA2993928A priority patent/CA2993928A1/en
Priority to BR112018001737A priority patent/BR112018001737A2/en
Priority to MX2018001223A priority patent/MX2018001223A/en
Priority to CN201680001947.1A priority patent/CN107466231A/en
Priority to EP16830853.4A priority patent/EP3328437A4/en
Application filed by Samyang Biopharmaceuticals Corporation filed Critical Samyang Biopharmaceuticals Corporation
Priority to JP2018504263A priority patent/JP2018521105A/en
Priority to AU2016299548A priority patent/AU2016299548A1/en
Publication of WO2017018820A1 publication Critical patent/WO2017018820A1/en
Priority to IL257187A priority patent/IL257187A/en
Priority to ZA2018/01187A priority patent/ZA201801187B/en
Priority to HK18109126.8A priority patent/HK1249441A1/en

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
    • 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
    • A61K31/12—Ketones
    • A61K31/122—Ketones having the oxygen directly attached to a ring, e.g. quinones, vitamin K1, anthralin
    • 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
    • A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
    • A61K31/337—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having four-membered rings, e.g. taxol
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
    • A61K47/08—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing oxygen, e.g. ethers, acetals, ketones, quinones, aldehydes, peroxides
    • A61K47/10—Alcohols; Phenols; Salts thereof, e.g. glycerol; Polyethylene glycols [PEG]; Poloxamers; PEG/POE alkyl ethers
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
    • A61K47/32—Macromolecular compounds obtained by reactions only involving carbon-to-carbon unsaturated bonds, e.g. carbomers, poly(meth)acrylates, or polyvinyl pyrrolidone
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
    • A61K47/34—Macromolecular compounds obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyesters, polyamino acids, polysiloxanes, polyphosphazines, copolymers of polyalkylene glycol or poloxamers
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/10—Dispersions; Emulsions
    • A61K9/107—Emulsions ; Emulsion preconcentrates; Micelles
    • A61K9/1075—Microemulsions or submicron emulsions; Preconcentrates or solids thereof; Micelles, e.g. made of phospholipids or block copolymers
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents

Definitions

  • the present disclosure relates to a pharmaceutical composition with improved storage stability and a method for preparing the same, and more specifically, a pharmaceutical composition of poorly water-soluble drug comprising an amphiphilic block copolymer wherein the content of a specific related compound is kept within a specified limit, and a method for preparing the same.
  • Solubilization of a poorly water-soluble drug is a key technology for delivering the drug into the body via oral or parenteral administration.
  • Such solubilization methods include a method of adding a surfactant to an aqueous solution to form micelles and then entrapping a poorly water-soluble drug therein.
  • An amphiphilic block copolymer used as a surfactant comprises a hydrophilic polymer block and a hydrophobic polymer block. Since the hydrophilic polymer block directly contacts blood proteins and cell membranes in vivo , polyethylene glycol or monomethoxypolyethylene glycol, etc. having biocompatibility has been used.
  • the hydrophobic polymer block improves affinity to a hydrophobic drug
  • polylactide, polyglycolide, poly(lactic-glycolide), polycaprolactone, polyamino acid or polyorthoester, etc. having biodegradability has been used.
  • polylactide derivatives have been applied to drug carriers in various forms because they have excellent biocompatibility and are hydrolyzed into harmless lactic acid in vivo .
  • Polylactide derivatives have various physical properties depending on their molecular weights, and have been developed in various forms such as microsphere, nanoparticle, polymeric gel and implant agent.
  • US Patent No. 6,322,805 discloses a composition for delivering a poorly water-soluble drug consisting of a polymeric micelle-type drug carrier and a poorly water-soluble drug, wherein the polymeric micelle-type drug carrier is formed from a diblock or triblock copolymer which is not crosslinked by a crosslinking agent and consists of at least one biodegradable hydrophobic polymer selected from the group consisting of polylactide, polyglycolide, poly(lactide-glycolide), polycaprolactone and derivatives thereof and poly(alkylene oxide) as a hydrophilic polymer, wherein the poorly water-soluble drug is physically entrapped in the drug carrier and solubilized, and wherein the polymeric micelle-type drug carrier forms a clear aqueous solution in water and effectively delivers the poorly water-soluble drug into the body.
  • the polymeric micelle-type drug carrier is formed from a diblock or triblock copolymer which is not crosslinked by a crosslinking agent and consists
  • polyethylene glycol-polylactide diblock copolymer is synthesized by removing moisture from monomethoxypolyethylene glycol, adding stannous octoate dissolved in toluene thereto and removing toluene under reduced pressure, adding D,L-lactide to the resulting mixture and conducting a polymerization reaction, adding chloroform to dissolve the produced block copolymer, dropwise adding an excess amount of diethyl ether in small portions with stirring to form precipitant and filtering the formed precipitant, and washing it several times with diethyl ether.
  • this method is difficult to employ in mass-scale production and thus is not commercially available.
  • the ether that has been used for purification may remain in the final polymeric micelle composition.
  • US Patent No. 8,853,351 discloses a method for preparing an amphiphilic block copolymer, comprising (a) dissolving the amphiphilic block copolymer in a water-miscible organic solvent; (b) adding and mixing an aqueous solution of alkali metal salt (sodium bicarbonate, sodium carbonate, potassium bicarbonate, potassium carbonate or lithium carbonate) to the polymeric solution obtained in step (a); (c) separating organic and aqueous phases by salting out for the solution obtained in step (b); and, (d) isolating the organic phase obtained in step (c) and removing the organic solvent therefrom to recover the polymer.
  • the method involves complicated steps, and requires an additional step for removing the alkali metal salt and the salt (sodium chloride or potassium chloride) used for salting out, and may have residual metal salts even after the removal thereof.
  • Impurities of drug must be strictly controlled in various aspects. Particularly, in case of impurities derived from active pharmaceutical ingredient (API), each country determines in its drug approval guideline the upper limit to amount of API-derived, known or unknown impurities (related compounds) in a drug product. In addition, there are some standards used internationally and ICH guideline Q3A is the representative one. In this guideline, at the time of approving a drug, the amount of each related compound in the drug is limited up to 0.1% or 0.2%, etc. and information such as toxicity-related data, etc., which should be provided, is discriminately applied according to the related compound exceeding the limit.
  • API active pharmaceutical ingredient
  • One purpose of the present invention is to provide a polymeric micelle-type pharmaceutical composition of poorly water-soluble drug comprising an amphiphilic block copolymer, which contains a specific related compound in an amount within a specified limit.
  • the other purpose of the present invention is to provide a method for preparing said pharmaceutical composition.
  • One aspect of the present invention provides a polymeric micelle pharmaceutical composition, comprising: a purified amphiphilic block copolymer comprising a hydrophilic block (A) and a hydrophobic block (B), and one or more poorly water-soluble drugs selected from the group consisting of paclitaxel and docetaxel, wherein the pharmaceutical composition contains, when stored at 40°C for 6 months, a related compound represented by the following Formula 1 in an amount of less than 0.12 part by weight, based on 100 parts by weight of the initial amount of the poorly water-soluble drug:
  • R 1 is H or COCH 3
  • R 2 is phenyl or O(CH 3 ) 3 .
  • Another aspect of the present invention provides a method for preparing a polymeric micelle pharmaceutical composition, comprising: (a) purifying an amphiphilic block copolymer comprising a hydrophilic block (A) and a hydrophobic block (B); (b) dissolving one or more poorly water-soluble drugs selected from the group consisting of paclitaxel and docetaxel, and the purified amphiphilic block copolymer in an organic solvent; and (c) adding an aqueous solvent to the solution obtained in step (b) to form polymeric micelles; wherein the pharmaceutical composition contains, when stored at 40°C for 6 months, a related compound represented by the above Formula 1 in an amount of less than 0.12 part by weight, based on 100 parts by weight of the initial amount of the poorly water-soluble drug.
  • a pharmaceutical composition of poorly water-soluble drug which has reduced related compounds and improved storage stability, can be obtained.
  • Figure 1 is the resulting chromatogram of HPLC analysis for the polymeric micelle composition containing paclitaxel used in Experimental Example 1, which had been subjected to the six-month acceleration test.
  • Figure 2 shows the results of product ion scan in LC/MS/MS analysis for the related compound (RRT 1.44 ⁇ 0.05 (1.39 ⁇ 1.49), with which RRT 1.44 is interchangeably used hereinafter) obtained in Experimental Example.
  • Figure 3 shows the results of LC/MS/MS analysis for the material obtained at RRT 1.44 position in the mixture obtained by thermally decomposing paclitaxel in Experimental Example 3.
  • Figure 4 shows the results of product ion scan in the LC/MS/MS analysis for the material obtained at RRT 1.44 position in the mixture obtained by thermally decomposing paclitaxel in Experimental Example 3, together with the analysis results of the six-month acceleration tested sample of the polymeric micelle composition:
  • Figure 5 shows the results of 1 H NMR analysis in the NMR analysis for the material obtained at RRT 1.44 position in the mixture obtained by thermally decomposing paclitaxel in Experimental Example 3.
  • Figure 6 shows the results of 13 C NMR analysis in the NMR analysis for the material obtained at RRT 1.44 position in the mixture obtained by thermally decomposing paclitaxel in Experimental Example 3.
  • Figure 7 shows the results of COSY (Correlation Spectroscopy) analysis in the NMR analysis for the material obtained at RRT 1.44 position in the mixture obtained by thermally decomposing paclitaxel in Experimental Example 3.
  • Figure 8 shows the results of HMBC (Heteronuclear Multiple Bond Correlation Spectroscopy) analysis in the NMR analysis for the material obtained at RRT 1.44 position in the mixture obtained by thermally decomposing paclitaxel in Experimental Example 3.
  • HMBC Heteronuclear Multiple Bond Correlation Spectroscopy
  • Figure 9 is the resulting chromatogram of HPLC analysis conducted in Experimental Example 6.
  • the pharmaceutical composition of an embodiment of the present invention comprises a purified amphiphilic block copolymer comprising a hydrophilic block (A) and a hydrophobic block (B).
  • the amphiphilic block copolymer comprises an A-B type diblock copolymer consisting of a hydrophilic block (A) and a hydrophobic block (B), or a B-A-B type triblock copolymer.
  • the amphiphilic block copolymer may comprise the hydrophilic block in an amount of 20 to 95% by weight, and more concretely 40 to 95% by weight, based on the total weight of the copolymer.
  • the amphiphilic block copolymer may comprise the hydrophobic block in an amount of 5 to 80% by weight, and more concretely 5 to 60% by weight, based on the total weight of the copolymer.
  • the amphiphilic block copolymer may have a number average molecular weight of 1,000 to 50,000 Daltons, and more concretely 1,500 to 20,000 Daltons.
  • the hydrophilic block is a polymer having biocompatibility and may comprise one or more selected from the group consisting of polyethylene glycol or derivatives thereof, polyvinylpyrrolidone, polyvinyl alcohol, polyacrylamide and combinations thereof, and more concretely, it may comprise one or more selected from the group consisting of polyethylene glycol, monomethoxypolyethylene glycol and combinations thereof.
  • the hydrophilic block may have a number average molecular weight of 200 to 20,000 Daltons, and more concretely 200 to 10,000 Daltons.
  • the hydrophobic block is a polymer having biodegradability and may be a polymer of monomers derived from alpha ( ⁇ )-hydroxy acid. Concretely, it may comprise one or more selected from the group consisting of polylactide, polyglycolide, polymandelic acid, polycaprolactone, polydioxan-2-one, polyamino acid, polyorthoester, polyanhydride, polycarbonate and combinations thereof, and more concretely, it may comprise one or more selected from the group consisting of polylactide, polyglycolide, polycaprolactone, polydioxan-2-one and combinations thereof.
  • the hydrophobic block may have a number average molecular weight of 200 to 20,000 Daltons, and more concretely 200 to 10,000 Daltons.
  • an amphiphilic block copolymer comprising a hydrophobic polymer block of poly(alpha ( ⁇ )-hydroxy acid) may be synthesized by a known ring-opening polymerization method using a hydrophilic polymer having hydroxyl group as an initiator, and a lactone monomer of alpha ( ⁇ )-hydroxy acid.
  • L-lactide or D,L-lactide may be polymerized with hydrophilic polyethylene glycol or monomethoxypolyethylene glycol having hydroxyl group as an initiator by ring-opening. Synthesis of diblock or triblock copolymer is possible according to the number of hydroxyl group existing in the hydrophilic block which is the initiator.
  • an organometallic catalyst such as tin oxide, lead oxide, tin octoate, antimony octoate, etc. may be used, and tin octoate having biocompatibility is preferably used in preparing polymer for medical use.
  • amphiphilic block copolymer a purified one is used.
  • the amphiphilic block copolymer is one that has been purified by sublimation.
  • the purification by sublimation may be conducted at a temperature of preferably 80°C or higher and lower than 120°C and more preferably 80 to 100°C, and under a pressure of a vacuum degree of preferably 10 torr or less, more preferably 5 torr or less and even more preferably 1 torr or less, for a time of preferably 10 to 74 hours, more preferably 10 to 48 hours and even more preferably 24 to 48 hours. Conducting the purification by sublimation under such conditions can minimize the change in molecular weight of the copolymer and remove impurities therefrom.
  • the pharmaceutical composition of an embodiment of the present invention comprises, as active ingredient, one or more poorly water-soluble drugs selected from the group consisting of paclitaxel and docetaxel.
  • the pharmaceutical composition may further comprise, as additional active ingredient, one or more poorly water-soluble drugs other than paclitaxel and docetaxel.
  • one or more taxane anticancer agents selected from the group consisting of 7-epipaclitaxel, t-acetylpaclitaxel, 10-desacetylpaclitaxel, 10-desacetyl-7-epipaclitaxel, 7-xylosylpaclitaxel, 10-desacetyl-7-glutarylpaclitaxel, 7-N,N-dimethylglycylpaclitaxel, 7-L-alanylpaclitaxel and cabazitaxel, may be used.
  • the pharmaceutical composition of an embodiment of the present invention may comprise the poorly water-soluble drug in an amount of 0.1 to 50 parts by weight, and more concretely 0.5 to 30 parts by weight, based on 100 parts by weight of the amphiphilic block copolymer. If the amount of the poorly water-soluble drug is too small as compared with that of the amphiphilic block copolymer, the weight ratio of the amphiphilic copolymer used per drug is high and thus the time for reconstitution may increase. On the other hand, if the amount of the poorly water-soluble drug is too large, there may be a problem of rapid precipitation of the poorly water-soluble drug.
  • the "initial" amount of the poorly water-soluble drug means the weight of the poorly water-soluble drug incorporated when the pharmaceutical composition was prepared.
  • the pharmaceutical composition contains, when stored at the accelerated condition (40°C) for 6 months, a related compound represented by the following Formula 1 in an amount of less than 0.12 part by weight, based on 100 parts by weight of the initial amount of the poorly water-soluble drug:
  • R 1 is H or COCH 3
  • R 2 is phenyl or O(CH 3 ) 3 .
  • the poorly water-soluble drug is paclitaxel
  • the related compound(s) may include the compound represented by the following Formula 1a:
  • the pharmaceutical composition of an embodiment of the present invention may contain, when stored at the accelerated condition (40°C) for 6 months, a related compound of Formula 1 (particularly, Formula 1a) in an amount of less than 0.12 part by weight, preferably 0.1 part by weight or less, more preferably 0.06 part by weight or less, even more preferably 0.05 part by weight or less, and most preferably 0.04 part by weight or less, based on 100 parts by weight of the initial amount of the poorly water-soluble drug.
  • a related compound of Formula 1 particularly, Formula 1a in an amount of less than 0.12 part by weight, preferably 0.1 part by weight or less, more preferably 0.06 part by weight or less, even more preferably 0.05 part by weight or less, and most preferably 0.04 part by weight or less, based on 100 parts by weight of the initial amount of the poorly water-soluble drug.
  • the pharmaceutical composition of an embodiment of the present invention may contain, when stored at the severe condition (80°C) for 3 weeks, a related compound of Formula 1 (particularly, Formula 1a) in an amount of less than 0.93 part by weight, preferably 0.8 part by weight or less, more preferably 0.6 part by weight or less, even more preferably 0.4 part by weight or less, and most preferably 0.2 part by weight or less, based on 100 parts by weight of the initial amount of the poorly water-soluble drug.
  • the pharmaceutical composition which contains a specific related compound in an amount within a specified limit, is a commercially available composition since it can be produced on a large scale.
  • the pharmaceutical composition of the present invention does not have ether, for example, diethyl ether, at all.
  • the pharmaceutical composition of the present invention does not have metal salt, for example, alkali metal salt and/or salt for salting out, for example, NaCl or KCl, at all.
  • metal salt for example, alkali metal salt and/or salt for salting out, for example, NaCl or KCl, at all.
  • the pharmaceutical composition of an embodiment of the present invention can be prepared by a method comprising (a) purifying an amphiphilic block copolymer comprising a hydrophilic block (A) and a hydrophobic block (B); (b) dissolving one or more poorly water-soluble drugs selected from the group consisting of paclitaxel and docetaxel, and the purified amphiphilic block copolymer in an organic solvent; and (c) adding an aqueous solvent to the solution obtained in step (b) to form polymeric micelles.
  • amphiphilic block copolymer The purification of the amphiphilic block copolymer is explained above, and a conventional method can be used for the formation of the polymeric micelles.
  • a water-miscible organic solvent for example, selected from the group consisting of alcohol (for example, ethanol), acetone, tetrahydrofuran, acetic acid, acetonitrile and dioxane and combinations thereof can be used, but it is not limited thereto.
  • aqueous solvent one selected from the group consisting of conventional water, distilled water, distilled water for injection, physiological saline, 5% glucose, buffer and combinations thereof can be used, but it is not limited thereto.
  • the method for preparing a pharmaceutical composition of an embodiment of the present invention may further comprise removing an organic solvent after said step (a).
  • the method may further comprise lyophilizing the micelle composition with addition of a lyophilization aid.
  • the lyophilization aid may be added for the lyophilized composition to maintain a cake form.
  • the lyophilization aid may be one or more selected from the group consisting of sugar and sugar alcohol.
  • the sugar may be one or more selected from lactose, maltose, sucrose or trehalose.
  • the sugar alcohol may be one or more selected from mannitol, sorbitol, maltitol, xylitol and lactitol.
  • the lyophilization aid may also function to facilitate homogeneous dissolution of the lyophilized polymeric micelle composition upon reconstitution.
  • the lyophilization aid may be contained at an amount of 1 to 90 weight%, particularly, 1 to 60 weight%, more particularly 10 to 60 weight%, based in a total weight of the lyophilized composition.
  • Preparation Example 1 Synthesis of diblock copolymer consisting of monomethoxypolyethylene glycol and D,L - lactide (mPEG- PDLLA ) and purification by sublimation method
  • mPEG monomethoxypolyethylene glycol
  • the reactor was tightly sealed and the polymerization reaction was conducted at 120°C for 10 hours. After the reaction was terminated, under agitation with a magnetic bar, the reactor was connected to a vacuum pump and the product was purified under a pressure of 1 torr or less by a sublimation method for 7 hours to obtain 262 g of mPEG-PDLLA in molten state.
  • the molecular weight (Mn: ⁇ 3740) was calculated by analyzing with 1 H-NMR obtaining relative intensities of appropriate peaks with reference to -OCH 3 which is the terminal group of monomethoxypolyethylene glycol.
  • Preparation Example 5 Purification of diblock copolymer (mPEG- PDLLA ) by adsorption method using aluminum oxide ( Al 2 O 3 )
  • a lyophilization aid 2.5 g of anhydrous lactose was added thereto and dissolved completely, filtered using a filter with a pore size of 200 nm, and freeze-dried to obtain a polymeric micelle composition containing paclitaxel in powder form.
  • Example 1 Preparation of polymeric micelle composition containing paclitaxel
  • a polymeric micelle composition containing paclitaxel was prepared by the same method as in Comparative Example 1.
  • a polymeric micelle composition containing paclitaxel was prepared by the same method as in Comparative Example 1.
  • Nebulizer gas pressure: Nitrogen (35 psi)
  • the substance for analysis which was isolated and came out of the detection stage, was set to flow in the mass spectrometer, and at that time the detected ion of related compound was qualitatively analyzed selecting the characteristic ion of mass spectrum [M+H].
  • Nebulizer gas pressure: Nitrogen (35 psi)
  • the material obtained at RRT 1.44 position from the mixture obtained by thermally decomposing paclitaxel in Experimental Example 3 was analyzed by NMR.
  • NMR analysis the results of 1 H NMR analysis are shown in Figure 5, the results of 13 C NMR analysis are shown in Figure 6, the results of COSY (Correlation Spectroscopy) analysis are shown in Figure 7, and the results of HMBC (Heteronuclear Multiple Bond Correlation Spectroscopy) analysis are shown in Figure 8.
  • the material obtained at RRT 1.44 position from the mixture obtained by thermally decomposing paclitaxel i.e., the related compound (RRT: 1.44 ⁇ 0.05 (1.39 ⁇ 1.49)) in the polymeric micelle composition containing paclitaxel which had been subjected to the six-month acceleration test
  • the related compound RRT: 1.44 ⁇ 0.05 (1.39 ⁇ 1.49)
  • the polymeric micelle composition containing paclitaxel which had been subjected to the six-month acceleration test was the compound of the following water-eliminated form of paclitaxel.
  • NMR equipment Brucker DRX-300 equipped with a temperature controller
  • Probe head Brucker 5mm QNP
  • the polymeric micelle compositions of paclitaxel prepared in Comparative Example 1 and Examples 1 and 2 were kept in an oven at 80°C for 3 weeks, and the compositions were then analyzed with HPLC to compare the amounts of related compound.
  • the test solution was prepared by dissolving the micelle composition in 80% acetonitrile aqueous solution and diluting to 600 ppm concentration of paclitaxel.
  • the resulting chromatogram of HPLC analysis is shown in Figure 9 and the change in the amount of related compound (%) according to the severe test time is shown in the following Table 2.
  • Amount of each related compound (%) 100(Ri/Ru)
  • the above test result shows an average value of the amounts of each related compound and paclitaxel in the test conducted for 3 or more polymeric micelle compositions of different batches.

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Abstract

A pharmaceutical composition containing a specific related compound in an amount within a specified limit and a method for preparing the same are provided.

Description

PHARMACEUTICAL COMPOSITION WITH IMPROVED STORAGE STABILITY AND METHOD FOR PREPARING THE SAME
The present disclosure relates to a pharmaceutical composition with improved storage stability and a method for preparing the same, and more specifically, a pharmaceutical composition of poorly water-soluble drug comprising an amphiphilic block copolymer wherein the content of a specific related compound is kept within a specified limit, and a method for preparing the same.
Solubilization of a poorly water-soluble drug is a key technology for delivering the drug into the body via oral or parenteral administration. Such solubilization methods include a method of adding a surfactant to an aqueous solution to form micelles and then entrapping a poorly water-soluble drug therein. An amphiphilic block copolymer used as a surfactant comprises a hydrophilic polymer block and a hydrophobic polymer block. Since the hydrophilic polymer block directly contacts blood proteins and cell membranes in vivo, polyethylene glycol or monomethoxypolyethylene glycol, etc. having biocompatibility has been used. The hydrophobic polymer block improves affinity to a hydrophobic drug, and polylactide, polyglycolide, poly(lactic-glycolide), polycaprolactone, polyamino acid or polyorthoester, etc. having biodegradability has been used. In particular, polylactide derivatives have been applied to drug carriers in various forms because they have excellent biocompatibility and are hydrolyzed into harmless lactic acid in vivo. Polylactide derivatives have various physical properties depending on their molecular weights, and have been developed in various forms such as microsphere, nanoparticle, polymeric gel and implant agent.
US Patent No. 6,322,805 discloses a composition for delivering a poorly water-soluble drug consisting of a polymeric micelle-type drug carrier and a poorly water-soluble drug, wherein the polymeric micelle-type drug carrier is formed from a diblock or triblock copolymer which is not crosslinked by a crosslinking agent and consists of at least one biodegradable hydrophobic polymer selected from the group consisting of polylactide, polyglycolide, poly(lactide-glycolide), polycaprolactone and derivatives thereof and poly(alkylene oxide) as a hydrophilic polymer, wherein the poorly water-soluble drug is physically entrapped in the drug carrier and solubilized, and wherein the polymeric micelle-type drug carrier forms a clear aqueous solution in water and effectively delivers the poorly water-soluble drug into the body. According to the above US patent, polyethylene glycol-polylactide diblock copolymer is synthesized by removing moisture from monomethoxypolyethylene glycol, adding stannous octoate dissolved in toluene thereto and removing toluene under reduced pressure, adding D,L-lactide to the resulting mixture and conducting a polymerization reaction, adding chloroform to dissolve the produced block copolymer, dropwise adding an excess amount of diethyl ether in small portions with stirring to form precipitant and filtering the formed precipitant, and washing it several times with diethyl ether. However, this method is difficult to employ in mass-scale production and thus is not commercially available. In addition, the ether that has been used for purification may remain in the final polymeric micelle composition.
US Patent No. 8,853,351 discloses a method for preparing an amphiphilic block copolymer, comprising (a) dissolving the amphiphilic block copolymer in a water-miscible organic solvent; (b) adding and mixing an aqueous solution of alkali metal salt (sodium bicarbonate, sodium carbonate, potassium bicarbonate, potassium carbonate or lithium carbonate) to the polymeric solution obtained in step (a); (c) separating organic and aqueous phases by salting out for the solution obtained in step (b); and, (d) isolating the organic phase obtained in step (c) and removing the organic solvent therefrom to recover the polymer. However, the method involves complicated steps, and requires an additional step for removing the alkali metal salt and the salt (sodium chloride or potassium chloride) used for salting out, and may have residual metal salts even after the removal thereof.
Impurities of drug must be strictly controlled in various aspects. Particularly, in case of impurities derived from active pharmaceutical ingredient (API), each country determines in its drug approval guideline the upper limit to amount of API-derived, known or unknown impurities (related compounds) in a drug product. In addition, there are some standards used internationally and ICH guideline Q3A is the representative one. In this guideline, at the time of approving a drug, the amount of each related compound in the drug is limited up to 0.1% or 0.2%, etc. and information such as toxicity-related data, etc., which should be provided, is discriminately applied according to the related compound exceeding the limit. This implies that since it is unknown how a related compound of a drug would act in vivo, the amount of the related compound must be reduced in the procedure of manufacturing the drug. Therefore, a manufacturing process for reducing the related compounds and setting of the upper limit to amount according to the characteristics (structure and toxicity) of each related compound are essential factors in quality control of the drug.
One purpose of the present invention is to provide a polymeric micelle-type pharmaceutical composition of poorly water-soluble drug comprising an amphiphilic block copolymer, which contains a specific related compound in an amount within a specified limit.
The other purpose of the present invention is to provide a method for preparing said pharmaceutical composition.
One aspect of the present invention provides a polymeric micelle pharmaceutical composition, comprising: a purified amphiphilic block copolymer comprising a hydrophilic block (A) and a hydrophobic block (B), and one or more poorly water-soluble drugs selected from the group consisting of paclitaxel and docetaxel, wherein the pharmaceutical composition contains, when stored at 40°C for 6 months, a related compound represented by the following Formula 1 in an amount of less than 0.12 part by weight, based on 100 parts by weight of the initial amount of the poorly water-soluble drug:
[Formula 1]
Figure PCTKR2016008270-appb-I000001
wherein
R1 is H or COCH3, and R2 is phenyl or O(CH3)3.
Another aspect of the present invention provides a method for preparing a polymeric micelle pharmaceutical composition, comprising: (a) purifying an amphiphilic block copolymer comprising a hydrophilic block (A) and a hydrophobic block (B); (b) dissolving one or more poorly water-soluble drugs selected from the group consisting of paclitaxel and docetaxel, and the purified amphiphilic block copolymer in an organic solvent; and (c) adding an aqueous solvent to the solution obtained in step (b) to form polymeric micelles; wherein the pharmaceutical composition contains, when stored at 40°C for 6 months, a related compound represented by the above Formula 1 in an amount of less than 0.12 part by weight, based on 100 parts by weight of the initial amount of the poorly water-soluble drug.
According to the present invention, a pharmaceutical composition of poorly water-soluble drug, which has reduced related compounds and improved storage stability, can be obtained.
Figure 1 is the resulting chromatogram of HPLC analysis for the polymeric micelle composition containing paclitaxel used in Experimental Example 1, which had been subjected to the six-month acceleration test.
Figure 2 shows the results of product ion scan in LC/MS/MS analysis for the related compound (RRT 1.44±0.05 (1.39~1.49), with which RRT 1.44 is interchangeably used hereinafter) obtained in Experimental Example.
Figure 3 shows the results of LC/MS/MS analysis for the material obtained at RRT 1.44 position in the mixture obtained by thermally decomposing paclitaxel in Experimental Example 3.
Figure 4 shows the results of product ion scan in the LC/MS/MS analysis for the material obtained at RRT 1.44 position in the mixture obtained by thermally decomposing paclitaxel in Experimental Example 3, together with the analysis results of the six-month acceleration tested sample of the polymeric micelle composition:
(a) Results of analysis of the six-month acceleration tested sample of the polymeric micelle pharmaceutical composition
(b) Results of analysis of the material obtained at RRT 1.44 position in the mixture obtained by thermally decomposing paclitaxel
Figure 5 shows the results of 1H NMR analysis in the NMR analysis for the material obtained at RRT 1.44 position in the mixture obtained by thermally decomposing paclitaxel in Experimental Example 3.
Figure 6 shows the results of 13C NMR analysis in the NMR analysis for the material obtained at RRT 1.44 position in the mixture obtained by thermally decomposing paclitaxel in Experimental Example 3.
Figure 7 shows the results of COSY (Correlation Spectroscopy) analysis in the NMR analysis for the material obtained at RRT 1.44 position in the mixture obtained by thermally decomposing paclitaxel in Experimental Example 3.
Figure 8 shows the results of HMBC (Heteronuclear Multiple Bond Correlation Spectroscopy) analysis in the NMR analysis for the material obtained at RRT 1.44 position in the mixture obtained by thermally decomposing paclitaxel in Experimental Example 3.
Figure 9 is the resulting chromatogram of HPLC analysis conducted in Experimental Example 6.
The present invention is explained in more detail below.
The pharmaceutical composition of an embodiment of the present invention comprises a purified amphiphilic block copolymer comprising a hydrophilic block (A) and a hydrophobic block (B).
According to one embodiment of the present invention, the amphiphilic block copolymer comprises an A-B type diblock copolymer consisting of a hydrophilic block (A) and a hydrophobic block (B), or a B-A-B type triblock copolymer.
According to one embodiment of the present invention, the amphiphilic block copolymer may comprise the hydrophilic block in an amount of 20 to 95% by weight, and more concretely 40 to 95% by weight, based on the total weight of the copolymer. In addition, the amphiphilic block copolymer may comprise the hydrophobic block in an amount of 5 to 80% by weight, and more concretely 5 to 60% by weight, based on the total weight of the copolymer.
According to one embodiment of the present invention, the amphiphilic block copolymer may have a number average molecular weight of 1,000 to 50,000 Daltons, and more concretely 1,500 to 20,000 Daltons.
According to one embodiment of the present invention, the hydrophilic block is a polymer having biocompatibility and may comprise one or more selected from the group consisting of polyethylene glycol or derivatives thereof, polyvinylpyrrolidone, polyvinyl alcohol, polyacrylamide and combinations thereof, and more concretely, it may comprise one or more selected from the group consisting of polyethylene glycol, monomethoxypolyethylene glycol and combinations thereof. The hydrophilic block may have a number average molecular weight of 200 to 20,000 Daltons, and more concretely 200 to 10,000 Daltons.
According to one embodiment of the present invention, the hydrophobic block is a polymer having biodegradability and may be a polymer of monomers derived from alpha (α)-hydroxy acid. Concretely, it may comprise one or more selected from the group consisting of polylactide, polyglycolide, polymandelic acid, polycaprolactone, polydioxan-2-one, polyamino acid, polyorthoester, polyanhydride, polycarbonate and combinations thereof, and more concretely, it may comprise one or more selected from the group consisting of polylactide, polyglycolide, polycaprolactone, polydioxan-2-one and combinations thereof. The hydrophobic block may have a number average molecular weight of 200 to 20,000 Daltons, and more concretely 200 to 10,000 Daltons.
According to one embodiment of the present invention, an amphiphilic block copolymer comprising a hydrophobic polymer block of poly(alpha (α)-hydroxy acid) may be synthesized by a known ring-opening polymerization method using a hydrophilic polymer having hydroxyl group as an initiator, and a lactone monomer of alpha (α)-hydroxy acid. For example, L-lactide or D,L-lactide may be polymerized with hydrophilic polyethylene glycol or monomethoxypolyethylene glycol having hydroxyl group as an initiator by ring-opening. Synthesis of diblock or triblock copolymer is possible according to the number of hydroxyl group existing in the hydrophilic block which is the initiator. In the ring-opening polymerization, an organometallic catalyst such as tin oxide, lead oxide, tin octoate, antimony octoate, etc. may be used, and tin octoate having biocompatibility is preferably used in preparing polymer for medical use.
In an embodiment of the present invention, as the amphiphilic block copolymer, a purified one is used. According to a preferable embodiment of the present invention, the amphiphilic block copolymer is one that has been purified by sublimation.
The purification by sublimation may be conducted at a temperature of preferably 80°C or higher and lower than 120°C and more preferably 80 to 100°C, and under a pressure of a vacuum degree of preferably 10 torr or less, more preferably 5 torr or less and even more preferably 1 torr or less, for a time of preferably 10 to 74 hours, more preferably 10 to 48 hours and even more preferably 24 to 48 hours. Conducting the purification by sublimation under such conditions can minimize the change in molecular weight of the copolymer and remove impurities therefrom.
The pharmaceutical composition of an embodiment of the present invention comprises, as active ingredient, one or more poorly water-soluble drugs selected from the group consisting of paclitaxel and docetaxel.
According to one embodiment of the present invention, the pharmaceutical composition may further comprise, as additional active ingredient, one or more poorly water-soluble drugs other than paclitaxel and docetaxel. As such an additional active ingredient, one or more taxane anticancer agents selected from the group consisting of 7-epipaclitaxel, t-acetylpaclitaxel, 10-desacetylpaclitaxel, 10-desacetyl-7-epipaclitaxel, 7-xylosylpaclitaxel, 10-desacetyl-7-glutarylpaclitaxel, 7-N,N-dimethylglycylpaclitaxel, 7-L-alanylpaclitaxel and cabazitaxel, may be used.
The pharmaceutical composition of an embodiment of the present invention may comprise the poorly water-soluble drug in an amount of 0.1 to 50 parts by weight, and more concretely 0.5 to 30 parts by weight, based on 100 parts by weight of the amphiphilic block copolymer. If the amount of the poorly water-soluble drug is too small as compared with that of the amphiphilic block copolymer, the weight ratio of the amphiphilic copolymer used per drug is high and thus the time for reconstitution may increase. On the other hand, if the amount of the poorly water-soluble drug is too large, there may be a problem of rapid precipitation of the poorly water-soluble drug.
As used herein, the "initial" amount of the poorly water-soluble drug means the weight of the poorly water-soluble drug incorporated when the pharmaceutical composition was prepared.
In an embodiment of the present invention, the pharmaceutical composition contains, when stored at the accelerated condition (40°C) for 6 months, a related compound represented by the following Formula 1 in an amount of less than 0.12 part by weight, based on 100 parts by weight of the initial amount of the poorly water-soluble drug:
[Formula 1]
Figure PCTKR2016008270-appb-I000002
wherein
R1 is H or COCH3, and R2 is phenyl or O(CH3)3.
According to one embodiment of the present invention, the poorly water-soluble drug is paclitaxel, and the related compound(s) may include the compound represented by the following Formula 1a:
[Formula 1a]
Figure PCTKR2016008270-appb-I000003
The pharmaceutical composition of an embodiment of the present invention may contain, when stored at the accelerated condition (40°C) for 6 months, a related compound of Formula 1 (particularly, Formula 1a) in an amount of less than 0.12 part by weight, preferably 0.1 part by weight or less, more preferably 0.06 part by weight or less, even more preferably 0.05 part by weight or less, and most preferably 0.04 part by weight or less, based on 100 parts by weight of the initial amount of the poorly water-soluble drug.
The pharmaceutical composition of an embodiment of the present invention may contain, when stored at the severe condition (80°C) for 3 weeks, a related compound of Formula 1 (particularly, Formula 1a) in an amount of less than 0.93 part by weight, preferably 0.8 part by weight or less, more preferably 0.6 part by weight or less, even more preferably 0.4 part by weight or less, and most preferably 0.2 part by weight or less, based on 100 parts by weight of the initial amount of the poorly water-soluble drug.
In an embodiment of the present invention, the pharmaceutical composition, which contains a specific related compound in an amount within a specified limit, is a commercially available composition since it can be produced on a large scale.
In an embodiment, the pharmaceutical composition of the present invention does not have ether, for example, diethyl ether, at all.
In an embodiment, the pharmaceutical composition of the present invention does not have metal salt, for example, alkali metal salt and/or salt for salting out, for example, NaCl or KCl, at all.
The pharmaceutical composition of an embodiment of the present invention can be prepared by a method comprising (a) purifying an amphiphilic block copolymer comprising a hydrophilic block (A) and a hydrophobic block (B); (b) dissolving one or more poorly water-soluble drugs selected from the group consisting of paclitaxel and docetaxel, and the purified amphiphilic block copolymer in an organic solvent; and (c) adding an aqueous solvent to the solution obtained in step (b) to form polymeric micelles.
The purification of the amphiphilic block copolymer is explained above, and a conventional method can be used for the formation of the polymeric micelles.
In the method for preparing a pharmaceutical composition of an embodiment of the present invention, as the organic solvent, a water-miscible organic solvent, for example, selected from the group consisting of alcohol (for example, ethanol), acetone, tetrahydrofuran, acetic acid, acetonitrile and dioxane and combinations thereof can be used, but it is not limited thereto. In addition, as the aqueous solvent, one selected from the group consisting of conventional water, distilled water, distilled water for injection, physiological saline, 5% glucose, buffer and combinations thereof can be used, but it is not limited thereto.
The method for preparing a pharmaceutical composition of an embodiment of the present invention may further comprise removing an organic solvent after said step (a).
In an embodiment, the method may further comprise lyophilizing the micelle composition with addition of a lyophilization aid. The lyophilization aid may be added for the lyophilized composition to maintain a cake form. In another embodiment, the lyophilization aid may be one or more selected from the group consisting of sugar and sugar alcohol. The sugar may be one or more selected from lactose, maltose, sucrose or trehalose. The sugar alcohol may be one or more selected from mannitol, sorbitol, maltitol, xylitol and lactitol. The lyophilization aid may also function to facilitate homogeneous dissolution of the lyophilized polymeric micelle composition upon reconstitution. The lyophilization aid may be contained at an amount of 1 to 90 weight%, particularly, 1 to 60 weight%, more particularly 10 to 60 weight%, based in a total weight of the lyophilized composition.
The present invention is explained in more detail by the following examples. However, these examples seek to illustrate the present invention only, and the scope of the present invention is not limited by the examples in any manner.
[EXAMPLES]
Preparation Example 1: Synthesis of diblock copolymer consisting of monomethoxypolyethylene glycol and D,L - lactide (mPEG- PDLLA ) and purification by sublimation method
150 g of monomethoxypolyethylene glycol (mPEG, number average molecular weight = 2,000) was fed into a 500-ml round-bottom flask equipped with an agitator, and agitated at 120°C under vacuum condition for 2 hours to remove moisture. 0.15 g of tin octoate (Sn(Oct)2) dissolved in 200 μl of toluene was added in the reaction flask, and further agitated under vacuum condition for 1 hour to distill and remove toluene. 150 g of D,L-lactide was then added and agitated under nitrogen atmosphere for dissolution. After D,L-lactide was dissolved completely, the reactor was tightly sealed and the polymerization reaction was conducted at 120°C for 10 hours. After the reaction was terminated, under agitation with a magnetic bar, the reactor was connected to a vacuum pump and the product was purified under a pressure of 1 torr or less by a sublimation method for 7 hours to obtain 262 g of mPEG-PDLLA in molten state. The molecular weight (Mn: ~3740) was calculated by analyzing with 1H-NMR obtaining relative intensities of appropriate peaks with reference to -OCH3 which is the terminal group of monomethoxypolyethylene glycol.
Preparation Example 2: Purification of diblock copolymer (mPEG- PDLLA ) by sublimation method
30 g of mPEG-PDLLA, which was obtained in the polymerization reaction process of Preparation Example 1 before conducting the purification process, was fed into a one-necked flask and dissolved at 80°C. Under agitation with a magnetic bar, the reactor was connected to a vacuum pump and the product was purified under a pressure of 1 torr or less by a sublimation method for 24 hours and 48 hours.
Preparation Example 3: Purification of diblock copolymer (mPEG- PDLLA ) by sublimation method
Except that the purification temperature was 100°C, the purification was conducted by the same method as in Preparation Example 2.
Preparation Example 4: Purification of diblock copolymer (mPEG- PDLLA ) by sublimation method
Except that the purification temperature was 120°C, the purification was conducted by the same method as in Preparation Example 2.
Preparation Example 5: Purification of diblock copolymer (mPEG- PDLLA ) by adsorption method using aluminum oxide ( Al 2 O 3 )
30 g of mPEG-PDLLA, which was obtained in the polymerization reaction process of Preparation Example 1 before conducting the purification process, was fed into a one-necked flask and dissolved by adding acetone (60 ml). Aluminum oxide (15 g) was added thereto and completely mixed. The one-necked flask was connected to a rotary evaporator, and the contents were mixed at 50°C at 60 rpm for 2 hours. The solution was then filtered at room temperature with PTFE filter paper (1 μm) to remove aluminum oxide. The filtered acetone solution was distilled using a rotary evaporator at 60°C under vacuum to remove acetone, thereby to obtain the purified mPEG-PDLLA. The molecular weight (Mn: ~3690) was calculated by analyzing with 1H-NMR obtaining relative intensities of appropriate peaks with reference to -OCH3 which is the terminal group of monomethoxypolyethylene glycol.
The molecular weight change of mPEG-PDLLA according to the purification conditions in the above Preparation Examples 2 to 5 is shown in the following Table 1.
[Table 1]
Figure PCTKR2016008270-appb-I000004
From the results of Table 1, it can be seen that the reduced amount of the molecular weight of mPEG-PDLLA increases as the purification temperature becomes higher. The purification condition of 80 to 100°C and 24 to 48 hours, particularly 100°C and 24 hours, can be thought of as efficient.
Comparative Example 1: Preparation of polymeric micelle composition containing paclitaxel
1 g of paclitaxel and 5 g of mPEG-PDLLA obtained in Preparation Example 1 were weighed, and 4 ml of ethanol was added thereto and agitated at 60°C until the mixture was completely dissolved to form a clear solution. Ethanol was then removed by distillation under reduced pressure using a rotary evaporator equipped with a round-bottom flask at 60°C for 3 hours. The temperature was then lowered to 50°C, and 140 ml of distilled water at room temperature was added and reacted until the solution became clear in blue color to form polymeric micelles. As a lyophilization aid, 2.5 g of anhydrous lactose was added thereto and dissolved completely, filtered using a filter with a pore size of 200 nm, and freeze-dried to obtain a polymeric micelle composition containing paclitaxel in powder form.
Example 1: Preparation of polymeric micelle composition containing paclitaxel
Except that mPEG-PDLLA purified for 24 hours in Preparation Example 3 was used, a polymeric micelle composition containing paclitaxel was prepared by the same method as in Comparative Example 1.
Example 2: Preparation of polymeric micelle composition containing paclitaxel
Except that mPEG-PDLLA purified in Preparation Example 5 was used, a polymeric micelle composition containing paclitaxel was prepared by the same method as in Comparative Example 1.
Experimental Example 1: Isolation of related compound by liquid chromatography
To a vial containing 100 mg of polymeric micelle composition containing paclitaxel, which had been subjected to the six-month acceleration test (temperature: 40°C), 16.7 ml of deionized water (DW) was fed and the contents were completely dissolved, and the total amount of the liquid was taken and transferred to a 20-ml volumetric flask, and the marked line was met to make the total volume 20 ml (5.0 mg/ml). 2 ml of this liquid was taken and transferred to a 10-ml volumetric flask, and the marked line was met with acetonitrile to make the total volume 10 ml (1 mg/ml). For the above composition, related compound was isolated and fractionally collected using the following liquid chromatography.
Conditions for liquid chromatography
1) Column: Poroshell 120 PFP (4.6 × 150 mm, 2.7 μm, Agilent)
2) Mobile phase: A: DW / B: Acetonitrile
Figure PCTKR2016008270-appb-I000005
3) Flow rate: 0.6 ml/min
4) Injection volume: 10 μl
5) Detector: UV absorption spectrophotometer (Measurement wavelength: 227 nm)
The resulting chromatogram of HPLC analysis is shown in Figure 1.
Experimental Example 2: Qualitative analysis of related compound of RRT 1.44 using LC/MS/MS
The related compound isolated in Experimental Example 1 (RRT: 1.44±0.05 (1.39~1.49)) was qualitatively analyzed by MS scan of liquid chromatography-mass spectrometer (LC/MS/MS). In the following measurement, as the LC/MS/MS, liquid chromatography 1200 series and electrospray ionization mass spectrometer 6400 series (Agilent, US) were used. The conditions for analysis were as follows.
Conditions for liquid chromatography
1) Column: Cadenza HS-C18 (3.0 × 150 mm, 3 μm, Imtakt)
2) Mobile phase: A: 0.5 mM ammonium acetate with 0.03% acetic acid / B: Acetonitrile
Figure PCTKR2016008270-appb-I000006
3) Flow rate: 0.4 ml/min
4) Injection volume: 2 μl
5) Detector: UV absorption spectrophotometer (Measurement wavelength: 227 nm)
Conditions for electrospray ionization mass spectrometer
1) Ionization: Electrospray Ionization, Positive (ESI+)
2) MS Method: MS2 scan / Product ion scan
3) Ion source: Agilent Jet Stream ESI
4) Nebulizer gas (pressure): Nitrogen (35 psi)
5) Ion spray voltage: 3500 V
6) Drying gas temperature (flow rate): 350°C (7 L/min)
7) Sheath gas temperature (flow rate): 400°C (10 L/min)
8) Fragmentor: 135 V
9) Nozzle voltage: 500 V
10) Cell accelerator voltage: 7 V
11) EMV: 0 V
12) Collision energy: 22 V
13) Precursor ion: m/z 836.2
14) Mass scan range: m/z 100~1500
The substance for analysis, which was isolated and came out of the detection stage, was set to flow in the mass spectrometer, and at that time the detected ion of related compound was qualitatively analyzed selecting the characteristic ion of mass spectrum [M+H].
Experimental Example 3: Thermal decomposition test of paclitaxel
In the related compounds which were fractionally collected from the polymeric nanoparticle composition containing paclitaxel in Experimental Example 1, many polymers existed together and thus direct experiment was very difficult. As a result of the qualitative analysis in the preliminary experiment using LC/MS/MS, the related compound was presumed as compounds produced by the elimination of water from paclitaxel. Accordingly, as a method of eliminating water molecule, an experiment of heating paclitaxel was carried out to confirm whether the presumed related compound was produced. First, 1 g of paclitaxel was vacuum-dried at 170°C for 2~3 hours and dissolved completely in 45 ml of acetonitrile, and 5 ml of DW was then added thereto. By using this solution, the related compound of RRT 1.44 was isolated and fractionally collected on prep-LC.
Experimental Example 4: Analysis of the related compound at RRT 1.44 position produced after the thermal decomposition reaction of paclitaxel using LC/MS/MS
The related compound fractionally collected in Experimental Example 3 (RRT: 1.44±0.05 (1.39~1.49)) was analyzed by liquid chromatography and mass spectrometer (LC/MS/MS). According to the HPLC analysis results, the material fractionally collected in Experimental Example 3 showed an HPLC peak at the same position as that of the related compound of RRT 1.44 in the polymeric micelle composition (Figure 4). This material was further analyzed by LC/MS/MS. As a result of the MS scan first, m/z 836.3 amu which is [M+H]+ was shown (Figure 3). The product ion scan was then conducted and the results thereof were shown in Figure 4. The results of the related compound of RRT 1.44 formed in the polymeric nanoparticle composition containing paclitaxel, which had been subjected to the six-month acceleration test, were shown together. In conclusion, it could be confirmed that the material fractionally collected at RRT 1.44 after thermally decomposing paclitaxel in Experimental Example 3 was the compound having the same structure as that of the related compound at RRT 1.44 position after the six-month acceleration test of the polymeric micelle composition.
Conditions for liquid chromatography
1) Column: Poroshell 120 PFP (4.6 × 150 mm, 2.7 μm, Agilent)
2) Mobile phase: A: DW / B: Acetonitrile
Figure PCTKR2016008270-appb-I000007
3) Flow rate: 0.6 ml/min
4) Injection volume: 10 μl
5) Detector: UV absorption spectrophotometer (Measurement wavelength: 227 nm)
Conditions for electrospray ionization mass spectrometer
1) Ionization: Electrospray Ionization, Positive (ESI+)
2) MS Method: MS2 scan / Product ion scan
3) Ion source: Agilent Jet Stream ESI
4) Nebulizer gas (pressure): Nitrogen (35 psi)
5) Ion spray voltage: 3500 V
6) Drying gas temperature (flow rate): 350°C (7 L/min)
7) Sheath gas temperature (flow rate): 400°C (10 L/min)
8) Fragmentor: 135 V
9) Nozzle voltage: 500 V
10) Cell accelerator voltage: 7 V
11) EMV: 0 V
12) Collision energy: 22 V
13) Precursor ion: m/z 836.2
14) Mass scan range: m/z 100~1500
Experimental Example 5: NMR analysis of material obtained at RRT 1.44 position from the mixture obtained by thermally decomposing paclitaxel
The material obtained at RRT 1.44 position from the mixture obtained by thermally decomposing paclitaxel in Experimental Example 3 was analyzed by NMR. In the NMR analysis, the results of 1H NMR analysis are shown in Figure 5, the results of 13C NMR analysis are shown in Figure 6, the results of COSY (Correlation Spectroscopy) analysis are shown in Figure 7, and the results of HMBC (Heteronuclear Multiple Bond Correlation Spectroscopy) analysis are shown in Figure 8.
According to the analysis results, it could be confirmed that the material obtained at RRT 1.44 position from the mixture obtained by thermally decomposing paclitaxel (i.e., the related compound (RRT: 1.44±0.05 (1.39~1.49)) in the polymeric micelle composition containing paclitaxel which had been subjected to the six-month acceleration test) was the compound of the following water-eliminated form of paclitaxel.
Figure PCTKR2016008270-appb-I000008
One molecule of water-eliminated form of paclitaxel: C47H49NO13 (835.91 g/mol)
Conditions for Nuclear Magnetic Resonance spectroscopy
1. 1 H
1) NMR equipment: Brucker DRX-300 equipped with a temperature controller 　
2) Sample: 1~10mg sample/ 0.6mL chloroform-d in 5mm o.d. NMR tube (In all NMR experiments, the same sample was used)
3) Probe head: Brucker 5mm QNP
4) Proton 90° pulse: 11μsec.
5) Relaxation delay/ Number of scan: 4.0 sec/ 8
2. 13 C
1) Probe head: Brucker 5mm QNP
2) Carbon 90° pulse, acquisition time: 8μsec, 4.0 sec
3) Relaxation delay/ Number of scan: 0.5 sec/ 36,092
3. COSY
1) NMR equipment: Brucker DRX-300
2) Probe head: Brucker 5mm QNP
3) Pulse sequence: cosyqf45
4) Proton 90° pulse: 11μsec.
5) Relaxation delay/ Number of scan/ Number of experiments for w1: 1.2 sec/ 4/ 256
4. HMQC
1) NMR equipment: Brucker DRX-300
2) Probe head: Brucker 5mm QNP
3) Pulse sequence: inv4ph
4) Proton 90° pulse: 11μsec.
5) Relaxation delay/ Number of scan/ Number of experiments for w1: 1.2 sec/ 64/ 256
6) Temperature, 1/2(JCH): 300K, 3.5 msec
5. HMBC
1) NMR equipment: Brucker DRX-300
2) Probe head: Brucker 5mm QNP
3) Pulse sequence: inv4lplrndqf
4) Proton 90° pulse: 11μsec.
5) Relaxation delay/ Number of scan / Number of experiments for a1: 1.5 sec/ 256/ 256
6) Temperature/ 1/2(JCH): 300K/ 3.5 msec
6. DEPT
1) Pulse sequence: DEPT 135
2) Carbon 90° pulse, acquisition time: 8μsec, 4.0 sec
3) Relaxation delay/ Number of scan/ 1/2(JCH): 1.2 sec/ 5628/ 3.5 msec
Experimental Example 6: Comparative test of storage stability of polymeric micelle containing drug at severe condition ( 80°C )
The polymeric micelle compositions of paclitaxel prepared in Comparative Example 1 and Examples 1 and 2 were kept in an oven at 80°C for 3 weeks, and the compositions were then analyzed with HPLC to compare the amounts of related compound. The test solution was prepared by dissolving the micelle composition in 80% acetonitrile aqueous solution and diluting to 600 ppm concentration of paclitaxel. The resulting chromatogram of HPLC analysis is shown in Figure 9 and the change in the amount of related compound (%) according to the severe test time is shown in the following Table 2.
HPLC conditions
Column: Diameter 2.7 μm, poroshell 120PFP (4.6 × 150 mm, 2.7 μm) (Agilent column)
Mobile phase
Figure PCTKR2016008270-appb-I000009
Detector: UV absorption spectrophotometer (227 nm)
Flow rate: 0.6 ml/min
Amount of each related compound (%) = 100(Ri/Ru)
Ri: Area of each related compound detected in test solution analysis
Ru: Sum of all peak areas detected in test solution analysis
[Table 2]
Figure PCTKR2016008270-appb-I000010
* RRT 0.87±0.02: Paclitaxel, oxetane ring opened compound
RRT 0.96±0.02: Paclitaxel, oxetane ring opened compound
RRT 1.00: Paclitaxel
RRT 1.10±0.02: Paclitaxel, L-lactide reaction compound
RRT 1.12±0.02: Paclitaxel, D-lactide reaction compound
RRT 1.44±0.05: Paclitaxel, water eliminated compound
From Table 2 and Figure 9, it can be known that the stability of the polymeric micelle pharmaceutical composition of Example 1 or 2 was improved as compared with the composition of Comparative Example 1 and the reduction of paclitaxel amount was relatively smaller, whereby the effect of the drug contained in the composition can be maintained more stably.
Experimental Example 7: Comparative test of storage stability of polymeric micelle containing drug at accelerated condition ( 40°C )
Except that the polymeric micelle composition of paclitaxel prepared in Comparative Example 1 and Example 1 respectively was kept in a stability tester at 40°C for 6 months, the test was conducted by the same method as in Experimental Example 6. The change in the amount of related compound (%) according to the acceleration test time is shown in the following Table 3.
[Table 3]
Figure PCTKR2016008270-appb-I000011
The above test result shows an average value of the amounts of each related compound and paclitaxel in the test conducted for 3 or more polymeric micelle compositions of different batches.
Through Experimental Example 7, it has been proven that the polymeric micelle pharmaceutical composition of Example 1, when stored at the accelerated storage temperature (40°C) for 6 months, has lower amount of related compound than the composition of Comparative Example 1.

Claims (11)

  1. A polymeric micelle pharmaceutical composition, comprising:
    a purified amphiphilic block copolymer comprising a hydrophilic block (A) and a hydrophobic block (B), and
    one or more poorly water-soluble drugs selected from the group consisting of paclitaxel and docetaxel,
    wherein the pharmaceutical composition contains, when stored at 40°C for 6 months, a related compound represented by the following Formula 1 in an amount of less than 0.12 part by weight, based on 100 parts by weight of the initial amount of the poorly water-soluble drug:
    [Formula 1]
    Figure PCTKR2016008270-appb-I000012
    wherein
    R1 is H or COCH3, and R2 is phenyl or O(CH3)3.
  2. The pharmaceutical composition according to claim 1, wherein the compound of Formula 1 is the compound of the following Formula 1a:
    [Formula 1a]
    Figure PCTKR2016008270-appb-I000013
  3. The pharmaceutical composition according to claim 1, which contains the related compound of Formula 1 in an amount of 0.1 part by weight or less, based on 100 parts by weight of the initial amount of the poorly water-soluble drug.
  4. The pharmaceutical composition according to claim 3, which contains the related compound of Formula 1 in an amount of 0.05 part by weight or less, based on 100 parts by weight of the initial amount of the poorly water-soluble drug.
  5. The pharmaceutical composition according to claim 4, which contains the related compound of Formula 1 in an amount of 0.04 part by weight or less, based on 100 parts by weight of the initial amount of the poorly water-soluble drug.
  6. The pharmaceutical composition according to claim 1, which contains, when stored at 80°C for 3 weeks, the related compound of Formula 1 in an amount of less than 0.93 part by weight, based on 100 parts by weight of the initial amount of the poorly water-soluble drug.
  7. The pharmaceutical composition according to claim 1, wherein the hydrophilic block (A) comprises one or more selected from the group consisting of polyethylene glycol or derivatives thereof, polyvinylpyrrolidone, polyvinyl alcohol, polyacrylamide and combinations thereof.
  8. The pharmaceutical composition according to claim 1, wherein the hydrophobic block (B) comprises one or more selected from the group consisting of polylactide, polyglycolide, polymandelic acid, polycaprolactone, polydioxan-2-one, polyamino acid, polyorthoester, polyanhydride, polycarbonate and combinations thereof.
  9. The pharmaceutical composition according to claim 1, wherein the hydrophilic block (A) is polyethylene glycol or monomethoxypolyethylene glycol, and the hydrophobic block (B) is polylactide.
  10. The pharmaceutical composition according to claim 1, wherein the hydrophilic block (A) has a number average molecular weight of 200 to 20,000 Daltons, and the hydrophobic block (B) has a number average molecular weight of 200 to 20,000 Daltons.
  11. The pharmaceutical composition according to claim 1, wherein the amphiphilic block copolymer is one purified by sublimation at a temperature of 80°C or higher and lower than 120°C and under a pressure of a vacuum degree of 10 torr or less for 10 to 74 hours.
PCT/KR2016/008270 2015-07-28 2016-07-28 Pharmaceutical composition with improved storage stability and method for preparing the same WO2017018820A1 (en)

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BR112018001737A BR112018001737A2 (en) 2015-07-28 2016-07-28 pharmaceutical composition with polymeric micelle
MX2018001223A MX2018001223A (en) 2015-07-28 2016-07-28 Pharmaceutical composition with improved storage stability and method for preparing the same.
CN201680001947.1A CN107466231A (en) 2015-07-28 2016-07-28 With improved pharmaceutical composition of storage stability and preparation method thereof
EP16830853.4A EP3328437A4 (en) 2015-07-28 2016-07-28 Pharmaceutical composition with improved storage stability and method for preparing the same
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Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2003033592A1 (en) * 2001-10-18 2003-04-24 Samyang Corporation Polymeric micelle composition with improved stability
US20110251269A1 (en) * 2008-12-26 2011-10-13 Samyang Corporation Preparation method of polymeric micelles composition containing a poorly water-soluble drug
US8853351B2 (en) * 2007-12-31 2014-10-07 Samyang Biopharmaceuticals Corporation Highly pure amphiphilic copolymer comprising hydrophobic block from alpha-hydroxy acid and process for the preparation thereof

Family Cites Families (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6020507A (en) 1998-03-02 2000-02-01 Bristol-Myers Squibb Company Synthesis of paclitaxel from baccatin III by protection of the 7-hydroxyl of baccatin III using a strong base and an electrophile
NZ522442A (en) * 2000-05-12 2004-04-30 Samyang Corp Method for the preparation of polymeric micelle via phase separation of block copolymer
US7217770B2 (en) * 2000-05-17 2007-05-15 Samyang Corporation Stable polymeric micelle-type drug composition and method for the preparation thereof
WO2005037840A1 (en) 2003-10-16 2005-04-28 Mayne Pharma (Usa), Inc. Method for inverting the c2’ hydroxyl group of taxane esters
KR101024742B1 (en) * 2007-12-31 2011-03-24 주식회사 삼양사 Amphiphilic Block Copolymer Micelle Composition Containing Taxane and Manufacturing Process of The Same
US9801818B2 (en) * 2007-12-31 2017-10-31 Samyang Biopharmaceuticals Corporation Method for stabilizing amphiphilic block copolymer micelle composition containing poorly water-soluble drug

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2003033592A1 (en) * 2001-10-18 2003-04-24 Samyang Corporation Polymeric micelle composition with improved stability
US8853351B2 (en) * 2007-12-31 2014-10-07 Samyang Biopharmaceuticals Corporation Highly pure amphiphilic copolymer comprising hydrophobic block from alpha-hydroxy acid and process for the preparation thereof
US20110251269A1 (en) * 2008-12-26 2011-10-13 Samyang Corporation Preparation method of polymeric micelles composition containing a poorly water-soluble drug

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
CIUTARU, D. ET AL.: "A HPLC validated assay of paclitaxel's related impurities in pharmaceutical forms containing Cremophor EL.", JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS, vol. 34, 2004, pages 493 - 499, XP055350129 *
KUMAR, D. ET AL.: "Isolation and characterization of degradation impurities in docetaxel drug substance and its formulation.", JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS, vol. 43, 2007, pages 1228 - 1235, XP005901637 *
See also references of EP3328437A4 *

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