+

WO2003049699A2 - Nouveaux aldehydes de polyethyleneglycol monofonctionnels - Google Patents

Nouveaux aldehydes de polyethyleneglycol monofonctionnels Download PDF

Info

Publication number
WO2003049699A2
WO2003049699A2 PCT/US2002/039434 US0239434W WO03049699A2 WO 2003049699 A2 WO2003049699 A2 WO 2003049699A2 US 0239434 W US0239434 W US 0239434W WO 03049699 A2 WO03049699 A2 WO 03049699A2
Authority
WO
WIPO (PCT)
Prior art keywords
pag
formula
aldehyde
integer
molecular weight
Prior art date
Application number
PCT/US2002/039434
Other languages
English (en)
Other versions
WO2003049699A3 (fr
Inventor
Perry Rosen
Kwang Nho
Original Assignee
Sun Bio, Inc.
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Priority claimed from KR10-2001-0078244A external-priority patent/KR100488351B1/ko
Application filed by Sun Bio, Inc. filed Critical Sun Bio, Inc.
Priority to EP02792347A priority Critical patent/EP1507755A4/fr
Priority to AU2002357806A priority patent/AU2002357806A1/en
Publication of WO2003049699A2 publication Critical patent/WO2003049699A2/fr
Publication of WO2003049699A3 publication Critical patent/WO2003049699A3/fr

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08G—MACROMOLECULAR COMPOUNDS OBTAINED OTHERWISE THAN BY REACTIONS ONLY INVOLVING UNSATURATED CARBON-TO-CARBON BONDS
    • C08G65/00—Macromolecular compounds obtained by reactions forming an ether link in the main chain of the macromolecule
    • C08G65/02—Macromolecular compounds obtained by reactions forming an ether link in the main chain of the macromolecule from cyclic ethers by opening of the heterocyclic ring
    • C08G65/32—Polymers modified by chemical after-treatment
    • C08G65/329—Polymers modified by chemical after-treatment with organic compounds
    • C08G65/333—Polymers modified by chemical after-treatment with organic compounds containing nitrogen
    • C08G65/33396—Polymers modified by chemical after-treatment with organic compounds containing nitrogen having oxygen in addition to nitrogen
    • 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
    • A61K47/50—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 the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—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 the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/56—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 the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule
    • A61K47/59—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 the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyureas or polyurethanes
    • A61K47/60—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 the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyureas or polyurethanes the organic macromolecular compound being a polyoxyalkylene oligomer, polymer or dendrimer, e.g. PEG, PPG, PEO or polyglycerol
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08G—MACROMOLECULAR COMPOUNDS OBTAINED OTHERWISE THAN BY REACTIONS ONLY INVOLVING UNSATURATED CARBON-TO-CARBON BONDS
    • C08G65/00—Macromolecular compounds obtained by reactions forming an ether link in the main chain of the macromolecule
    • C08G65/02—Macromolecular compounds obtained by reactions forming an ether link in the main chain of the macromolecule from cyclic ethers by opening of the heterocyclic ring
    • C08G65/32—Polymers modified by chemical after-treatment
    • C08G65/321—Polymers modified by chemical after-treatment with inorganic compounds
    • C08G65/324—Polymers modified by chemical after-treatment with inorganic compounds containing oxygen
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08G—MACROMOLECULAR COMPOUNDS OBTAINED OTHERWISE THAN BY REACTIONS ONLY INVOLVING UNSATURATED CARBON-TO-CARBON BONDS
    • C08G65/00—Macromolecular compounds obtained by reactions forming an ether link in the main chain of the macromolecule
    • C08G65/02—Macromolecular compounds obtained by reactions forming an ether link in the main chain of the macromolecule from cyclic ethers by opening of the heterocyclic ring
    • C08G65/32—Polymers modified by chemical after-treatment
    • C08G65/329—Polymers modified by chemical after-treatment with organic compounds
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08G—MACROMOLECULAR COMPOUNDS OBTAINED OTHERWISE THAN BY REACTIONS ONLY INVOLVING UNSATURATED CARBON-TO-CARBON BONDS
    • C08G65/00—Macromolecular compounds obtained by reactions forming an ether link in the main chain of the macromolecule
    • C08G65/02—Macromolecular compounds obtained by reactions forming an ether link in the main chain of the macromolecule from cyclic ethers by opening of the heterocyclic ring
    • C08G65/32—Polymers modified by chemical after-treatment
    • C08G65/329—Polymers modified by chemical after-treatment with organic compounds
    • C08G65/331—Polymers modified by chemical after-treatment with organic compounds containing oxygen

Definitions

  • Therapeutic proteins which are generally administered by intravenous injection may be immunogenic, relatively water insoluble, and may have a short in vivo half -life.
  • the pharmacokinetics of the particular protein will govern both the efficacy and duration of effect of the drug. It has become of major importance to reduce the rate of clearance of the protein so that prolonged action can be achieved. This may be accomplished by avoiding or inhibiting glomerular filtration which can be effected both by the charge on the protein and its molecular size (Brenner et al., (1978) Am.J.Physiol.,234,F455).
  • 4,129,337 discloses conjugating PEG to proteins such as enzymes and insulin to produce a less immunogenic product while retaining a substantial proportion of the biological activity.
  • PEG modification requires activation of the PEG polymer which is accomplished by the introduction of an electrophilic center.
  • the PEG reagent is now susceptible to nucleophilic attack, predominantly by the nucleophilic epsilon-amino group of a lysyl residue. Because of the number of surface lysines present in most proteins, the PEGylation process can result in random attachments leading to mixtures which are difficult to purify and which may not be desirable for pharmaceutical use.
  • a desired property therefore of a new reagent would be one that is not susceptible to degradation in an aqueous medium and one which may be employed to affect the site specific modification of a protein.
  • a PEG aldehyde may be considered such a reagent.
  • For site specific N-terminal pegylation see Pepinsky et al., (2001) JPET,297,1059 (Interferon- ⁇ -la) and U.S.Pat.No.5,824,784(1998) to Kinstler et al., (G-CSF).
  • the use of a PEG-aldehyde for the reductive amination of a protein utilizing other available nucleophilic amino groups, is described in U.S.Pat.
  • R is hydrogen or lower alkyl
  • X and Y are individually selected from -O - or - NH- with the proviso that X is NH when m is 1 and Y is -O-
  • PAG is a divalent residue of polyalkylene glycol resulting from removal of the terminal hydroxy groups, having a molecular weight of from 1,000 to 100,000 Daltons, z is an integer of from 2 to 4, m is an integer of from 0 to 1, and w is an integer of from 2 to 8, preferably 2 to 4.
  • A is a polyethylene glycol residue with its two terminal hydroxy groups being removed having a molecular weight of from 1,000 to 100,000 Daltons and having a valence of from 1 to 5; n is an integer of from 1 to 5 which integer is the same as the valence of A; R and w are as above.
  • PAG 1 and PAG 2 are independently divalent residues of poly lower alkylene glycol resulting from removal of the two terminal hydroxy groups with the PAG 1 and PAG 2 residues having a combined molecular weight of from 1,000 to 100,000 Daltons; R and R 1 are individually lower alkyl or hydrogen and w is as above and p is an integer of from 1 to 5; and z is as above, are useful for conjugation to therapeutically active proteins to produce PAG Protein conjugates which retain a substantial portion of their therapeutic activity and are less immunogenic than the protein from which the conjugate is derived.
  • the aldehyde reagents of formula IA, IB, IC and ID can be conjugated to therapeutically active proteins to produce therapeutically active protein conjugates which retain a substantial portion of the biological activity of the protein from which they are derived.
  • the reagents of this invention are not susceptible to degradation in the aqueous medium in which the pegylation reaction is carried out.
  • the aldehyde reagents of this invention can be conjugated to the protein in a controlled manner at the N- terminus. In this way, these aldehydes produce the desired conjugates and avoid random attachment leading to mixtures which are difficult to purify and which may not be desirable for pharmaceutical use.
  • the therapeutic proteins which can be conjugated in accordance with this invention can be any of the conventional therapeutic proteins.
  • the preferred proteins are included interferon-alpha, interferon-beta, consensus interferon, G-CSF, GM-CSF, EPO, Hemoglobin, interleukins, colony stimulating factor, as well as immunoglobulins such as IgG, IgE, IgM, IgA, IgD and fragments thereof.
  • polyalkylene glycol designates poly(lower alkylene)glycol radicals where the alkylene radical is a straight or branched chain radical containing from 2 to 7 carbon atoms.
  • lower alkylene designates a straight or branched chain divalent alkylene radical containing from 2 to 7 carbon atoms such as polyethylene, polypropylene, poly n- butylene, and polyisobutylene as well as polyalkylene glycols formed from mixed alkylene glycols such as polymers containing a mixture of polyethylene and polypropylene radicals and polymers containing a mixture of polyisopropylene, polyethylene and polyisobutylene radicals.
  • the branched chain alkylene glycol radicals provide the lower alkyl groups in the polymer chain of from 2 to 4 carbon atoms depending on the number of carbon atoms contained in the straight chain of the alkylene group so that the total number of carbons atoms of any alkylene moiety which makes up the polyalkylene glycol substituent is from 2 to 7.
  • the term "lower alkyl” includes lower alkyl groups containing from 1 to 7 carbon atoms, preferably from 1 to 4 carbon atoms, such as methyl, ethyl, propyl, isopropyl, etc. with methyl being especially preferred.
  • PAG in the compound in formulas IA, IC and ID is a polyethylene glycol residue formed by removal of the two terminal hydroxy groups.
  • PAG in the compound of formula IA, IC and ID, and the A in the compound of formula IB have molecular weights of from about 10,000 to 50,000 most preferably from about 20,000 to about 40,000.
  • the radicals PAG 1 and PAG 2 have a combined molecular weight of from about 10,000 to 50,000 and most preferably from about 20,000 to 40,000.
  • p be an integer of from 1 to 5.
  • the aldehydes of compounds of formula IA, IB, IC and ID are used in forming poly alky leneoxy protein conjugates.
  • the aldehydes of this invention are intermediates for conjugation with the terminal amino group as well as other free amino groups on the protein to produce a therapeutically effective conjugate which has the therapeutic properties of the native protein.
  • the conjugates show a reduced rate of clearance and a decreased antigenicity as compared to that of the starting protein.
  • these conjugates have the beneficial properties of in vivo reduced proteolysis, increased water solubility, reduced renal clearance, and steric hindrance to receptor-mediated clearance. These enhanced properties when compared to the protein from which they are formed make them more effective therapeutic agents than the protein itself.
  • the aldehydes of this invention are converted to their protein conjugates in accordance with the following reaction scheme:
  • the compound of formula IV is a conventional hydrate of the aldehyde of formula V.
  • An equilibrium between the formulas IV and V is established when the compound of formula V is placed in an aqueous medium.
  • the polyalkylene aldehyde of formula V is then reacted with the amine of the protein to form the imine linkage of formula VI.
  • This imine linkage of the compound of formula VI is then reduced to an amine through the use of reducing agents such as cyanoborohydride to give the saturated conjugated protein of formula VII.
  • the reaction whereby aldehydes are conjugated with proteins through reductive amination is set forth in U.S. Patent No. 4,002,531, EPO 154,316 and U.S. Patent No. 5,824,784.
  • This can be done by carrying out the reaction of the compound of formula V with P-NH 2 at a pH of from 5.5 to 7.5.
  • various buffers which maintain the reaction media at a pH of from 5.5 to 7.5 can be used. If one wants the amination to proceed on more than one amino site on the protein, then one carries out the reaction at a pH of 8.0 and above, preferably at a pH of from 8 to 10. In this manner, amino groups, as well as, the N-terminal amino group on the protein are aminated with the PAG aldehydes of this invention.
  • the specific PEGylating reagents of formula IA, IB, IC and ID of this invention are stable in aqueous medium and not subject to aldol decompositions under the conditions of the reductive amination reaction.
  • the amino groups on proteins such as those on the lysine residues are the predominate nucleophilic centers for the condensation of the aldehydes of this invention.
  • the pH of the reaction one can produce a site specific introduction of a polyalkylene glycol polymer on the protein at the desired N- terminus amino acid.
  • the acid group of the compound in formula VIII is activated to produce the compound of formula IX. This is accomplished by activating the acid group on the compound of formula VIJI with an activating agent to produce a leaving group such as an N-hydroxy succinimide group. Any conventional method of converting a carboxy group into an activating leaving group such as an N-hydroxy succinimide group can be utilized to produce the compound of formula IX.
  • the compound of formula IX containing the activating leaving group is reacted with the amine acetal compound of formula X to produce the compound of formula XL
  • This reaction to form the amide of formula XI is carried out by any conventional means of condensing an amine with an activated carboxylic acid group.
  • the compound of formula XI has the aldehyde protected as its acetal, preferably a lower alkyl acetal. Any conventional aldehyde protecting groups such as other alkyl acetals can also be utilized.
  • the acetal of formula XI can be hydrolyzed to form the corresponding aldehyde of formula I-Ai.
  • the compound of formula I-Aii can be prepared by the following reaction scheme. o
  • the carbonate is then reacted witi the amine of formula X to produce the compound of formula XV.
  • This reaction is carried out as described hereinbefore with regard to reacting the compound of formula IX with the compound of formula X.
  • the compound of formula XV is then hydrolyzed to produce the compound of formula I-Aii in the conventional manner as described in connection with the hydrolysis of the compound of formula XI hereinbefore.
  • the compound of formula XVI is condensed with the compound of formula XVII in a halogenated hydrocarbon solvent to produce the compound of formula XVIII.
  • This reaction utilizes conventional condensing procedures commonly used in reactions between an activated carbonate and an amine.
  • the compound of formula XVHI is condensed with the amine of formula X in an inert organic solvent to produce the acetal of formula XLX. Any conventional inert organic solvent can be used in this reaction.
  • the acetal of formula XLX is then hydrolyzed in acidic medium, in the manner described hereinabove to produce the compound of formula I-Aiii.
  • the compound of formula IA where m is 1, Y is -NH- and X is -O- the compound has the following formula:
  • R, PAG, z and w are as above.
  • the starting material of formula XX is a tri-hydroxy compound having two terminal primary hydroxy groups with the third hydroxy group being a secondary hydroxy group, vicinal to the one of the two terminal hydroxy groups.
  • the compound of formula XX is converted to its acetonide derivative of formula XXI by reacting the two vicinal hydroxy groups with acetone leaving free the third hydroxy group. Any conventional method of forming an acetonide derivative from the two vicinal hydroxy groups can be utilized to carry out this reaction to form the compound of formula XXI. Reagents other than acetone, which are known to form cyclic acetals with 1,2-diols, may also be used.
  • the free hydroxy group in the acetonide derivative of formula XXI is then activated with an activating group such as the p-nitro phenyl chloro formate as is shown in the reaction scheme.
  • an activating group such as the p-nitro phenyl chloro formate as is shown in the reaction scheme.
  • This reaction to convert the hydroxy group into an activated derivative is well known in the art.
  • the compound of formula XXII is produced where the primary hydroxy group on the compound of formula XXI is activated.
  • the compound of formula XXII is then condensed with the PEG amine of formula XVI to form the condensation product of formula XXIII. Any conditions conventional in reacting an activated alcohol with an amine to produce a urethane can be utilized to carry out this condensation.
  • the compound of formula XXIII containing the acetonide is then cleaved utilizing conditions conventional in cleaving acetonides such as by treatment with a mild acid, to produce the corresponding di-hydroxy compound.
  • the resulting dihydroxy groups are then oxidized with mild oxidizing agents such as a periodate oxidizing agent to produce the aldehyde of formula I-Aiv. Any conventional method of oxidizing a vicinal di-hydroxy compound to the corresponding aldehyde can be utilized to carry out this conversion.
  • the compound of formula IB is synthesized from RO-PEG-OR by reaction with acrylic acid by the following reaction scheme:
  • the acrylic acid of formula XXV can be reacted with the polyethylene glycol polymer of formula XXIV in the manner disclosed in U.S. Patent 4,528,334 Knopf, et al. to produce the compound of formula XXVI.
  • the addition of acrylic acid across the various polyethylene glycol units in the series of polyethylene glycol residues designated A can be controlled so that from 1 to 5 bonds with the acrylic acid will take place to form the acrylic acid graft copolymer of formula XXVI. In this manner depending upon the conditions used, as disclosed in U.S. Patent 4,528,334, from 1 to 5 additions of acrylic acid will occur in the polyethyleneoxy chain.
  • an activated form of the carboxy group of the graft copolymer of formula XXVI is reacted with the compound of formula X to form the compound of formula XXVII via amide formation.
  • This reaction is carried out in the same manner as described hereinbefore in connection with the conversion of the compound of formula VIII to the compound of formula XI by reaction of the compound of formula X, through the use of an appropriate carboxy activating leaving group as in formula IX.
  • the acetal of formula XXVII can then be hydrolyzed to the compound of formula IB as described in connection with the conversion of the acetal of XI to the aldehyde of formula I-Ai.
  • the compound of formula IC can be prepared as shown by the following reaction scheme:
  • the derivative of formula IC is prepared from a compound of formula XXVIII by first activating the carboxyl group. This carboxyl group can be activated in the manner disclosed herein before with respect to the activation of the compound of the formula VIII to produce the compound of the formula LX.
  • the activated compound is then condensed with the amino acetal compound of formula X to produce the compound of formula XXLX in the same manner as described herein before in connection with the reaction of the compound of formula LX with the compound of formula X to produce the compound of formula XL
  • the compound of formula XXLX is next converted to the compound of the formula IC by acid hydrolysis as described herein before in connection with the preparation of the compound of formula I-Ai from compound XI.
  • the compound of formula LD is produced from a compound of the formula XII via the following reaction scheme:
  • RO-PAG-0(CH 2 ) z -OH Act ⁇ vatlon ⁇ RO-PAG-0(CH 2 ) z -X
  • X may be a halogen or sulfonate ester and B is an alkalai metal.
  • the compound of formula XII is converted to the compound of formula XXX by converting the hydroxy group on the compound of formula XII to an activating leaving group.
  • the conversion of the terminal hydroxyl group of compound XII into an activated halide leaving group X can be readily achieved by reaction with a conventional halogenating reagent such as thionyl bromide.
  • the hydroxy group of the compound of formula XXI may be converted to a sulfonate ester by reaction with a halide of the activating leaving group such as mesyl or tosyl chloride.
  • a halide of the activating leaving group such as mesyl or tosyl chloride.
  • Any conventional method for converting the hydroxy group of compound XII to an activating leaving group such as a tosylate or mesylate or any of the aforementioned leaving groups can be utilized to produce the compound of formula XXX. This reaction may be carried out by reacting the formula XII with a halide of an activating leaving group such as tosyl chloride.
  • the compound of formula XXX can then be condensed with the compound of formula XXI to form the compound of formula XXXI.
  • the acetonide group is a precursor to the aldehyde of formula ED.
  • the acetonide can be hydrolyzed in mild acid.
  • any conventional means to produce the resulting dihydroxy compound from an acetonide can be used in this conversion.
  • the dihydroxy compound resulting form this hydrolysis can then be oxidized with a periodate to give the aldehyde of formula LO.
  • This aldehyde can be reacted as set forth in Scheme 1 with a protein to form the conjugate of the compound of formula ID with the protein at the N-terminal amino acid of the protein as described hereinbefore.
  • the compound of formula LD can also be produced from a compound of the formula XXI via the following reaction scheme.
  • PEG is a divalent residue of polyethylene glycol resulting from removal of the terminal hydroxy groups, having a molecular weight of from 1,000 to 100,000 Daltons.
  • the compound of formula XXI is reacted with any conventional organic alkali metal base such as potassium naphthalide to form the corresponding alkoxide XXII.
  • Liquid ethylene oxide is then added under conventional polymeric conditions to a solution of XXII. In this manner the anionic ring opening and polymerization of ethylene oxide is allowed to proceed under conditions that are well known for the production of polyethylene glycol polymers.
  • the amount of polymerization of the ethylene oxide can be controlled by conventional means to produce a polyethylene polymer of any desired molecular weight. Any remaining ethylene oxide can then be removed from the reaction mixture and an excess of an alkyl halide such as methyl iodide reacted for several hours to form a terminal alkyl ether.
  • the product, the compound of formula XXXIH, can then be isolated and converted to a compound of the formula ED in the same manner as described herein before for the conversion of the compound of formula XXXI to the compound of formula ED.
  • the aldehydes of formula IA, IB, IC and ID can be conjugated as described herein before with various proteins through an amine group on the protein by the process of reductive amination as disclosed in U.S. Patent 5,824,784 dated October 20, 1998.
  • the aldehydes in this invention may condense at the N-terminus amino group of a protein so as to obtain a monoconjugate derivative.
  • the pegylating reagents of IA, EB, IC and ED can from site specific mono-conjugates with the N-terminal amino group of various proteins thereby avoiding the necessity of employing extensive purification or separation techniques.
  • the reductive amination procedure will also involve the various lysine amino groups which are available in the protein molecule.
  • preferred proteins for such conjugations are included G-CSF, GM-CSF, interferon- ⁇ , interferon- ⁇ , EPO and Hemoglobin.
  • the mPEG-ethyl acetate was dissolved in 1 N-sodium hydroxide and stirred for 15 hours at room temperature.
  • the reaction mixture was then adjusted to pH 2 with 1 N aqueous HC1 and extracted twice with dichloromethane.
  • the extracted organic layer was dried over magnesium sulfate and the organic solvent removed.
  • Diethyl ether was then added to the residue and the precipitated compound filtered.
  • the product was dried under vacuum and the resulting acid 3 obtained as a white powder.
  • the mPEG-succinimidyl acetate 4 was dissolved in dichloromethane and stirred at room temperature while a solution of l-amino-3, 3-diethoxypropane in dichloromethane was added. The resulting solution was then stirred for 2 hours at room temperature. Precipitation was induced by the addition of diethyl ether. The product was then filtered and recrystalized from ethyl acetate. The recrystalized compound was dried under vacuum to give 5 as a white powder.
  • the diethyl acetal 5 was dissolved in an aqueous solution containing phosphoric acid (pHl) and stirred for 2 hours at 40-50°C. After cooling the reaction mixture to room temperature, the acidity was reduced to a pH6 by the addition of a 5% aqueous sodium bicarbonate solution. Brine was added and the resulting mixture extracted twice with dichloromethane. The organic layer was dried over magnesium sulfate, filtered and the solvent evaporated under reduced pressure. Precipitation was induced by the addition of diethyl ether to the crude residue. The product was collected and dried under vacuum to give 6 as a white powder.
  • phosphoric acid pH6
  • Brine was added and the resulting mixture extracted twice with dichloromethane.
  • the organic layer was dried over magnesium sulfate, filtered and the solvent evaporated under reduced pressure. Precipitation was induced by the addition of diethyl ether to the crude residue.
  • the product was collected and dried under vacuum to give 6 as a white powder
  • the integer n may be from 22 to 2,300 but more preferably 22 to 1,000.
  • the integer n may be from 22 to 2,300 but more preferably 22 to 1,000.
  • the diethyl acetal 3 (5g) was dissolved in an aqueous solution containing phosphoric acid (pHl) and stirred for 2 hours at 40-50°C. After cooling the reaction mixture to room temperature, the acidity was reduced to a pH6 by the addition of a 5% aqueous sodium bicarbonate solution. Brine was added and the resulting mixture extracted twice with dichloromethane. The organic layer was dried over magnesium sulfate, filtered and the solvent evaporated under reduced pressure. Precipitation was induced by the addition of diethyl ether to the crude residue. The product was collected and dried under vacuum to give 4 as a white powder.
  • phosphoric acid pH6
  • Brine was added and the resulting mixture extracted twice with dichloromethane.
  • the organic layer was dried over magnesium sulfate, filtered and the solvent evaporated under reduced pressure. Precipitation was induced by the addition of diethyl ether to the crude residue.
  • the product was collected and dried under vacuum to give 4 as
  • the integer n may be from 22 to 2,300 but more preferably 22 to 1,000.
  • n may be from 22 to 2,300 but more preferably 22 to 1,000.
  • Example 5 Scheme E (Type I-Aiii) Synthesis of mPEG-urea-propionaldchyde.
  • n may be from 22 to 2,300 but more preferably 22 to 1,000.
  • Example 6 Scheme F (Type I-Aiv) Synthesis of mPEG-urethane-butyraldehyde.
  • the above urethane-acetonide of formula 5 (5g, 0.5mmol) was dissolved in 75ml of 0. IM HCl and stirred for 6h. The water and HCl were then removed under reduced pressure to give the corresponding diol product. To 5g of the diol dissolved in 75ml water was added 267mg of NaIO (1.25mmol) and the reaction allowed to proceed for 5h in the dark. The aldehyde of formula 6 was then isolated by size exclusion chromatography on a Sephadex G 10 column. Oxidation of the 1,2-diol may also be realized using NaIO 4 supported on wet silica gel. Using this procedure the aldehyde is obtained without hydrate fo ⁇ nation. (see Vo- Quang et al., (1989) Synthesis No.1,64).
  • the integer n may be from 22 to 2,300 but more preferably 22 to 1,000.
  • Nonane was added to a reaction vessel containing mPEG (M.W. 20,000), PEG (M.W. 20,000), or a dim-PEG and heated to 140-145°C.
  • acrylic acid and t-butyl peroxybenzoate (a reaction initiator) were slowly added to the reaction mixture over a period of 1.5 hours.
  • the mixture was stirred for an additional hour at 140-145°C.
  • methanol was added to the mixture and heated and stirred until a homogeneous solution was obtained. The hot solution was then filtered under vacuum and the filtrate diluted with a 90/10/ v/v MeOH/H 2 O solution.
  • the resulting mixture was then filtered through a Pall Filtron ultrafiltration system and the filtrate then concentrated under reduced pressure.
  • the residue was dissolved by heating with a 50/50 v/v acetone/isopropyl alcohol solution, cooled to room temperature, and placed in the refrigerator overnight.
  • the product 1 was then filtered, washed 3 times with 50/50 v/v acetone/isopropyl alcohol solution and finally 3 times with diethyl ether and then vacuum dried overnight.
  • the acid number of the pendant-PEG-propionic acid 1 was detennined (mg of KOH needed to neutralize one gram of sample).
  • the pendant-PEG-propionic acid 1 was dissolved in dichloromethane and cooled to 0- 5°C. N-hydroxysuccinimide was then added followed by the addition of dicyclohexylcarbodimide dissolved in chloromethane. After stirring for 15 hours at room temperature, the dicyclohexylurea by product was removed from the reaction mixture via filtration and the residual organic solvent evaporated under vacuum (see Example 2). The crude residue was recrystalized from ethyl acetate, filtered, washed twice with diethyl ether, and dried for 12 hours under vacuum to give the pendant PEG-succinimidyl propionate 2 as a white powder.
  • the pendant-PEG-propionaldehyde diethyl acetal 3 was dissolved in an aqueous solution containing phosphoric acid (pH 1) and stirred for 2 hours at 40-50°C. After cooling the reaction mixture to room temperature, the acidity was reduced to a pH6 by the addition of a 5% aqueous sodium bicarbonate solution. Brine was added and the resulting mixture extracted twice with dichloromethane. The organic layer was dried over magnesium sulfate, filtered and the solvent evaporated under reduced pressure. Precipitation was induced by the addition of diethyl ether to the crude residue. The product was collected and dried under vacuum to give the pendant PEG-amide propionaldehye 4 as a white powder.
  • phosphoric acid pH 1
  • Brine was added and the resulting mixture extracted twice with dichloromethane.
  • the organic layer was dried over magnesium sulfate, filtered and the solvent evaporated under reduced pressure. Precipitation was induced by the addition of diethyl ether to
  • the integer m may be from 22 to 2,300 but more preferably 22 to 1,000.
  • the integer n may be 1 to 20 and more preferably 1 to 5.
  • Scheme H (Type IC) Synthesis of Branched mPEG-amide-propionaldehyde. The conversion of the branched chain carboxy acid 1 to the corresponding propionaldehyde 2 was carried out as described in Example 7.
  • R, R 1 , PAG 1 , PAG 2 , p and z are as above.
  • the PEG acetonide 5 (3.5g) was dissolved in 40ml of 0.1M HCl and stirred for 6h. The water and HCl were then removed under reduced pressure to give the corresponding diol product. To 3g of the 1,2-diol dissolved in 40ml water ( ⁇ 0.3mmol of diol) was added 160mg of NaIO 4 (0.75mmol) and the reaction allowed to proceed for 5h in the dark to produce the compound of formula 6_which was isolated by size exclusion chromatography on a Sephadex G 10 column. By using the same procedure, compounds of the type ED can be prepared whereby the integer m may be from 22 to 23,000.
  • the integer n may be from 22 to 2,300 but more preferably 22 to 1,000.

Landscapes

  • Chemical & Material Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • General Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Polymers & Plastics (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Inorganic Chemistry (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Epidemiology (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Medicinal Preparation (AREA)
  • Polyethers (AREA)
  • Peptides Or Proteins (AREA)

Abstract

L'invention concerne un nouvel aldéhyde de polyéthylèneglycol monofonctionnel destiné à la pégylation de protéines thérapeutiquement actives en vue d'une production de conjugués protéiques pégylés conservant une partie substantielle de leur activité thérapeutique et présentant une immunogénicité inférieure à celle de la protéine dont le conjugué est issu. L'invention concerne également une nouvelle méthode de synthèse destinée à la préparation de ces aldéhydes.
PCT/US2002/039434 2001-12-11 2002-12-09 Nouveaux aldehydes de polyethyleneglycol monofonctionnels WO2003049699A2 (fr)

Priority Applications (2)

Application Number Priority Date Filing Date Title
EP02792347A EP1507755A4 (fr) 2001-12-11 2002-12-09 Nouveaux aldehydes de polyethyleneglycol monofonctionnels
AU2002357806A AU2002357806A1 (en) 2001-12-11 2002-12-09 Novel monofunctional polyethylene glycol aldehydes

Applications Claiming Priority (8)

Application Number Priority Date Filing Date Title
KR10-2001-0078244 2001-12-11
KR10-2001-0078244A KR100488351B1 (ko) 2001-12-11 2001-12-11 신규한 폴리에틸렌글리콜-프로피온알데히드 유도체
US34845202P 2002-01-16 2002-01-16
US60/348,452 2002-01-16
US38150302P 2002-05-17 2002-05-17
US60/381,503 2002-05-17
US40774102P 2002-09-03 2002-09-03
US60/407,741 2002-09-03

Publications (2)

Publication Number Publication Date
WO2003049699A2 true WO2003049699A2 (fr) 2003-06-19
WO2003049699A3 WO2003049699A3 (fr) 2004-12-29

Family

ID=27483540

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US2002/039434 WO2003049699A2 (fr) 2001-12-11 2002-12-09 Nouveaux aldehydes de polyethyleneglycol monofonctionnels

Country Status (3)

Country Link
EP (1) EP1507755A4 (fr)
AU (1) AU2002357806A1 (fr)
WO (1) WO2003049699A2 (fr)

Cited By (12)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2003061577A3 (fr) * 2002-01-18 2003-10-02 Biogen Inc Composes polymeres de polyalkylene et utilisations de ceux-ci
WO2004022630A2 (fr) * 2002-09-09 2004-03-18 Nektar Therapeutics Al, Corporation Alcanales polymeres solubles dans l'eau
WO2004060300A2 (fr) 2002-12-26 2004-07-22 Mountain View Pharmaceuticals, Inc. Conjugues polymeres de cytokines, de chimiomokines, de facteurs de croissance, d'hormones polypeptidiques et d'antagonistes de ceux-ci conservant une activite de liaison aux recepteurs
WO2005000360A2 (fr) * 2003-05-23 2005-01-06 Nektar Therapeutics Al, Corporation Derives polymeres ayant des agencements d'atomes particuliers
US7273909B2 (en) 2002-07-24 2007-09-25 Hoffmann-La Roche Inc. Polyethylene glycol aldehydes
WO2007149067A2 (fr) * 2005-05-12 2007-12-27 Zymogenetics, Inc. Procédés d'utilisation de phhla2 pour une stimulation conjointe des lymphocytes t
FR2920777A1 (fr) * 2007-09-11 2009-03-13 Centre Nat Rech Scient Procede de preparation de derives (poly(oxyde d'ethylene) poly(oxyde de propylene) thermosensibles utiles pour fonctionnaliser le chitosane
EP2070951A1 (fr) 2007-12-14 2009-06-17 Fresenius Kabi Deutschland GmbH Procédé de production d'un dérivé hydroxyalkyle de l'amidon avec deux liens
EP2070950A1 (fr) 2007-12-14 2009-06-17 Fresenius Kabi Deutschland GmbH Dérivés hydroxyalkylés de l'amidon et leur procédé de préparation
WO2010007626A1 (fr) * 2008-07-14 2010-01-21 Biocon Limited Procédé de synthèse d'un mélange sensiblement monodispersé d'oligomères
US7947261B2 (en) 2003-05-23 2011-05-24 Nektar Therapeutics Conjugates formed from polymer derivatives having particular atom arrangements
US9125880B2 (en) 2002-12-26 2015-09-08 Mountain View Pharmaceuticals, Inc. Polymer conjugates of interferon-beta with enhanced biological potency

Families Citing this family (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE10209821A1 (de) 2002-03-06 2003-09-25 Biotechnologie Ges Mittelhesse Kopplung von Proteinen an ein modifiziertes Polysaccharid
WO2005014655A2 (fr) 2003-08-08 2005-02-17 Fresenius Kabi Deutschland Gmbh Conjugues d'amidon d'hydroxyalkyle et de proteine

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5135683A (en) * 1991-03-28 1992-08-04 Arco Chemical Technology, L.P. Process for producing a deprotected alkoxylated polyol
WO2002059179A2 (fr) * 2000-12-18 2002-08-01 Shearwater Corporation Synthese de derives polymeres non peptidiques de poids moleculaire eleve
WO2003044056A2 (fr) * 2001-11-20 2003-05-30 Pharmacia Corporation Conjugues de l'hormone de croissance humaine chimiquement modifiee

Family Cites Families (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5252714A (en) * 1990-11-28 1993-10-12 The University Of Alabama In Huntsville Preparation and use of polyethylene glycol propionaldehyde
US5990237A (en) * 1997-05-21 1999-11-23 Shearwater Polymers, Inc. Poly(ethylene glycol) aldehyde hydrates and related polymers and applications in modifying amines
WO2000024697A1 (fr) * 1998-10-26 2000-05-04 University Of Utah Research Foundation Procede de preparation de derives aldehydes des polyethyleneglycols

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5135683A (en) * 1991-03-28 1992-08-04 Arco Chemical Technology, L.P. Process for producing a deprotected alkoxylated polyol
WO2002059179A2 (fr) * 2000-12-18 2002-08-01 Shearwater Corporation Synthese de derives polymeres non peptidiques de poids moleculaire eleve
WO2003044056A2 (fr) * 2001-11-20 2003-05-30 Pharmacia Corporation Conjugues de l'hormone de croissance humaine chimiquement modifiee

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
See also references of EP1507755A2 *

Cited By (37)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US8524660B2 (en) 2002-01-18 2013-09-03 Biogen Idec Ma Inc. Polyalkylene polymer compounds and uses thereof
WO2003061577A3 (fr) * 2002-01-18 2003-10-02 Biogen Inc Composes polymeres de polyalkylene et utilisations de ceux-ci
US8017733B2 (en) 2002-01-18 2011-09-13 Biogen Idec Ma Inc. Polyalkylene polymer compounds and uses thereof
EA009783B1 (ru) * 2002-01-18 2008-04-28 Байоджен Айдек Ма Инк. Полиалкиленгликоль с остатком для конъюгации биологически активного соединения
EA011829B1 (ru) * 2002-01-18 2009-06-30 Байоджен Айдек Ма Инк. Полиалкиленгликоль с остатком для конъюгации биологически активного соединения
US7273909B2 (en) 2002-07-24 2007-09-25 Hoffmann-La Roche Inc. Polyethylene glycol aldehydes
US7511094B2 (en) 2002-09-09 2009-03-31 Nektar Therapeutics Al, Corporation Water-soluble polymer alkanals
WO2004022630A3 (fr) * 2002-09-09 2004-04-15 Nektar Therapeutics Al Corp Alcanales polymeres solubles dans l'eau
US8853325B2 (en) 2002-09-09 2014-10-07 Nektar Therapeutics Water-soluble polymer alkanals
US8076412B2 (en) 2002-09-09 2011-12-13 Nektar Therapeutics Water-soluble polymer alkanals
US7157546B2 (en) 2002-09-09 2007-01-02 Nektar Therapeutics Al, Corporation Water-soluble polymer alkanals
US7838595B2 (en) 2002-09-09 2010-11-23 Nektar Therapeutics Water-soluble polymer alkanals
WO2004022630A2 (fr) * 2002-09-09 2004-03-18 Nektar Therapeutics Al, Corporation Alcanales polymeres solubles dans l'eau
EP1628618A4 (fr) * 2002-12-26 2009-09-09 Mountain View Pharmaceuticals Conjugues polymeres de cytokines, de chimiomokines, de facteurs de croissance, d'hormones polypeptidiques et d'antagonistes de ceux-ci conservant une activite de liaison aux recepteurs
WO2004060300A2 (fr) 2002-12-26 2004-07-22 Mountain View Pharmaceuticals, Inc. Conjugues polymeres de cytokines, de chimiomokines, de facteurs de croissance, d'hormones polypeptidiques et d'antagonistes de ceux-ci conservant une activite de liaison aux recepteurs
US9125880B2 (en) 2002-12-26 2015-09-08 Mountain View Pharmaceuticals, Inc. Polymer conjugates of interferon-beta with enhanced biological potency
EP1628618A2 (fr) * 2002-12-26 2006-03-01 Mountain View Pharmaceuticals, Inc. Conjugues polymeres de cytokines, de chimiomokines, de facteurs de croissance, d'hormones polypeptidiques et d'antagonistes de ceux-ci conservant une activite de liaison aux recepteurs
WO2005000360A2 (fr) * 2003-05-23 2005-01-06 Nektar Therapeutics Al, Corporation Derives polymeres ayant des agencements d'atomes particuliers
EA011351B1 (ru) * 2003-05-23 2009-02-27 Нектар Терапеутикс Ал, Корпорейшн Полимерные реагенты, способы их получения, а также содержащие их конъюгаты и фармацевтические препараты
US11446385B2 (en) 2003-05-23 2022-09-20 Nektar Therapeutics Polymer derivatives having particular atom arrangements
WO2005000360A3 (fr) * 2003-05-23 2005-07-28 Nektar Therapeutics Al Corp Derives polymeres ayant des agencements d'atomes particuliers
US10646578B2 (en) 2003-05-23 2020-05-12 Nektar Therapeutics Polymer derivatives having particular atom arrangements
US7887789B2 (en) 2003-05-23 2011-02-15 Nektar Therapeutics Polymer derivatives having particular atom arrangements
US7947261B2 (en) 2003-05-23 2011-05-24 Nektar Therapeutics Conjugates formed from polymer derivatives having particular atom arrangements
US9913913B2 (en) 2003-05-23 2018-03-13 Nektar Therapeutics Method for preparing a polymeric reagent
US9399070B2 (en) 2003-05-23 2016-07-26 Nektar Therapeutics Compositions comprising a polymeric reagent and a pharmacologically active agent
US9044516B2 (en) 2003-05-23 2015-06-02 Nektar Therapeutics Pharmaceutical compositions of conjugates formed from polymer derivatives having particular atom arrangements
US8633277B2 (en) 2003-05-23 2014-01-21 Nektar Therapeutics Methods for forming polymer-drug conjugates
WO2007149067A2 (fr) * 2005-05-12 2007-12-27 Zymogenetics, Inc. Procédés d'utilisation de phhla2 pour une stimulation conjointe des lymphocytes t
WO2007149067A3 (fr) * 2005-05-12 2008-07-31 Zymogenetics Inc Procédés d'utilisation de phhla2 pour une stimulation conjointe des lymphocytes t
WO2009034130A1 (fr) * 2007-09-11 2009-03-19 Centre National De La Recherche Scientifique (Cnrs) Procédé de préparation de dérivés (poly(oxyde d'éthylène) poly(oxyde de propylène)) thermosensibles utiles pour fonctionnaliser le chitosane.
FR2920777A1 (fr) * 2007-09-11 2009-03-13 Centre Nat Rech Scient Procede de preparation de derives (poly(oxyde d'ethylene) poly(oxyde de propylene) thermosensibles utiles pour fonctionnaliser le chitosane
US8377418B2 (en) 2007-09-11 2013-02-19 Centre National De La Recherche Scientifique (Cnrs) Process for preparing thermosensitive (poly(ethylene oxide) poly(propylene oxide)) derivatives that can be used to functionalize chitosan
EP2070950A1 (fr) 2007-12-14 2009-06-17 Fresenius Kabi Deutschland GmbH Dérivés hydroxyalkylés de l'amidon et leur procédé de préparation
EP2070951A1 (fr) 2007-12-14 2009-06-17 Fresenius Kabi Deutschland GmbH Procédé de production d'un dérivé hydroxyalkyle de l'amidon avec deux liens
US9040723B2 (en) 2008-07-14 2015-05-26 Biocon Limited Method of synthesizing a substantially monodispersed mixture of oligomers
WO2010007626A1 (fr) * 2008-07-14 2010-01-21 Biocon Limited Procédé de synthèse d'un mélange sensiblement monodispersé d'oligomères

Also Published As

Publication number Publication date
EP1507755A4 (fr) 2006-05-17
AU2002357806A1 (en) 2003-06-23
AU2002357806A8 (en) 2003-06-23
EP1507755A2 (fr) 2005-02-23
WO2003049699A3 (fr) 2004-12-29

Similar Documents

Publication Publication Date Title
US7504477B2 (en) Polyalkylene glycol acid additives
JP4344110B2 (ja) ヘテロ二官能性ポリエチレングリコール誘導体およびその調製方法
US20040115165A1 (en) Bifunctional polyethylene glycol derivatives
WO2003049699A2 (fr) Nouveaux aldehydes de polyethyleneglycol monofonctionnels
US6916962B2 (en) Monofunctional polyethylene glycol aldehydes
US10035880B2 (en) Heterobifunctional poly(ethylene glycol) derivatives and methods for their preparation
US6956135B2 (en) Monofunctional polyethylene glycol aldehydes
US20030153694A1 (en) Novel monofunctional polyethylene glycol aldehydes
US20040147687A1 (en) Novel monofunctional polyethylene glycol aldehydes
JP5150297B2 (ja) ヘテロ二官能性ポリエチレングリコール誘導体およびその調製方法
JP6361083B2 (ja) ヘテロ二官能性ポリエチレングリコール誘導体およびその調製方法
KR100480423B1 (ko) 에리트로포이에틴과 폴리에틸렌글리콜 유도체의 배합체
JP6500318B2 (ja) ヘテロ二官能性ポリエチレングリコール誘導体およびその調製方法
JP5953459B2 (ja) ヘテロ二官能性ポリエチレングリコール誘導体およびその調製方法
JP6184462B2 (ja) ヘテロ二官能性ポリエチレングリコール誘導体およびその調製方法
US20070141021A1 (en) Methylmaleimidyl polymer derivatives

Legal Events

Date Code Title Description
AK Designated states

Kind code of ref document: A2

Designated state(s): AE AG AL AM AT AU AZ BA BB BG BR BY BZ CA CH CN CO CR CU CZ DE DK DM DZ EC EE ES FI GB GD GE GH GM HR HU ID IL IN IS JP KE KG KP KR KZ LC LK LR LS LT LU LV MA MD MG MK MN MW MX MZ NO NZ OM PH PL PT RO RU SD SE SG SK SL TJ TM TN TR TT TZ UA UG UZ VN YU ZA ZM ZW

AL Designated countries for regional patents

Kind code of ref document: A2

Designated state(s): GH GM KE LS MW MZ SD SL SZ TZ UG ZM ZW AM AZ BY KG KZ MD RU TJ TM AT BE BG CH CY CZ DE DK EE ES FI FR GB GR IE IT LU MC NL PT SE SI SK TR BF BJ CF CG CI CM GA GN GQ GW ML MR NE SN TD TG

121 Ep: the epo has been informed by wipo that ep was designated in this application
WWE Wipo information: entry into national phase

Ref document number: 2002792347

Country of ref document: EP

WWP Wipo information: published in national office

Ref document number: 2002792347

Country of ref document: EP

NENP Non-entry into the national phase in:

Ref country code: JP

WWW Wipo information: withdrawn in national office

Country of ref document: JP

点击 这是indexloc提供的php浏览器服务，不要输入任何密码和下载