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WO2018177055A1 - Method for preparing y-branched hydrophilic polymer carboxylic acid derivative - Google Patents

Method for preparing y-branched hydrophilic polymer carboxylic acid derivative Download PDF

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Publication number
WO2018177055A1
WO2018177055A1 PCT/CN2018/077229 CN2018077229W WO2018177055A1 WO 2018177055 A1 WO2018177055 A1 WO 2018177055A1 CN 2018077229 W CN2018077229 W CN 2018077229W WO 2018177055 A1 WO2018177055 A1 WO 2018177055A1
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group
carboxylic acid
acid derivative
hydrophilic polymer
substituted
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PCT/CN2018/077229
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French (fr)
Chinese (zh)
Inventor
陈晓萌
林美娜
张如军
赵宣
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北京键凯科技股份有限公司
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Priority claimed from CN201710915311.4A external-priority patent/CN108659227B/en
Application filed by 北京键凯科技股份有限公司 filed Critical 北京键凯科技股份有限公司
Priority to EP18775559.0A priority Critical patent/EP3604388B1/en
Publication of WO2018177055A1 publication Critical patent/WO2018177055A1/en
Priority to US16/588,620 priority patent/US11359089B2/en

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    • 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
    • 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
    • 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
    • C08G69/00—Macromolecular compounds obtained by reactions forming a carboxylic amide link in the main chain of the macromolecule
    • 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
    • C08G69/00—Macromolecular compounds obtained by reactions forming a carboxylic amide link in the main chain of the macromolecule
    • C08G69/48—Polymers modified by chemical after-treatment

Definitions

  • the invention relates to the technical field of polymers, in particular to a method for preparing a Y-branched hydrophilic polymer carboxylic acid derivative, in particular to a high-purity high molecular weight Y-branched polyethylene glycol carboxylic acid derivative. Preparation.
  • PEG Polyethylene glycol
  • Y-type polyethylene glycol is a widely used polyethylene glycol, which can significantly reduce the loss of activity of modified drugs, especially in improving the clinical application of protein and peptide drugs. It prevents the antibody from approaching the protein drug, thereby greatly increasing the circulating half-life of the protein drug in the body while greatly reducing its immunogenicity in vivo.
  • the terminal group of the Y-type PEG when the terminal group of the Y-type PEG is a carboxyl group, it can react with an amino group, a hydroxyl group or a thiol group on a modified drug or other compound to form a covalent bond to realize a modified linkage, and is a commonly used Y-type PEG derivative.
  • the preparation method thereof is as described in the patent CN1243779C.
  • the adsorption strength of the above three compounds on the column is ranked as mPEG- Cm, mPEG-gly, Y-cm.
  • the above preparation reaction product includes the target product Y-cm and the unreacted reactants mPEG-gly and mPEG-cm.
  • mPEG-gly greatly interferes with the separation, so that the purity of the target product is The yield is lowered.
  • the yield of the Y-type PEG product is low, only 50%, the product separation is difficult, and the cost is high, which is not favorable for industrial amplification.
  • the present invention provides a process for the preparation of a Y-branched hydrophilic polymeric carboxylic acid derivative.
  • the method includes the following reaction:
  • P a and P b are the same or different hydrophilic polymer residues
  • X 1 and X 3 are a linking group independently selected from: -(CH 2 ) i -, a combination of one or more of -(CH 2 ) i O-, -(CH 2 ) i S- and -(CH 2 ) i CO-, i is an integer from 0 to 10,
  • X 2 is a linking group, chosen from :-( CH 2) r -, - (CH 2) r O -, - (CH 2) r S- , and a combination of one or more of the following, r is an integer from 0 to 10,
  • F is a terminal group selected from: substituted or unsubstituted C 1-6 alkyl, C 1-6 substituted or unsubstituted alkoxy group,
  • R 1 and R 2 are independently selected from: -H, C 1-6 substituted or unsubstituted alkyl, C 1-6 substituted or unsubstituted alkoxy, C 3-6 substituted or unsubstituted cycloalkyl And a C 4-10 substituted or unsubstituted alkylene cycloalkyl group,
  • R 3 is selected from the group consisting of: -H, a C 1-6 substituted or unsubstituted alkyl group, a C 6-10 substituted or unsubstituted aralkyl group, and a C 4-10 substituted or unsubstituted heterocycloalkyl group,
  • the above preparation method is more suitable for preparing a higher molecular weight Y-branched hydrophilic polymer carboxylic acid derivative, and the reaction yield and product purity are higher, and separation is easier.
  • the Y-branched hydrophilic polymer carboxylic acid derivative may have a molecular weight of 15 to 50 KDa (specifically 15, 16, 16, 20, 22, 24, 26, 28) , 30, 32, 34, 36, 38, 40, 42, 44, 46, 48 or 50 KDa).
  • the anhydride is an organic acid anhydride.
  • the organic acid anhydride is selected from the group consisting of: di-tert-butyl dicarbonate (Boc anhydride), acetic anhydride, propionic anhydride, isobutyric anhydride, butyric anhydride, benzoic anhydride, and phthalic anhydride. One or more of them.
  • the anhydride is a Boc anhydride.
  • the anhydride and reactant The molar ratio of the added amount is 0.01-10:1 (specifically, 0.01:1, 0.1:1, 0.2:1, 0.3:1, 0.4:1, 0.5:1, 0.6:1, 0.7:1, 0.8:1) , 0.9:1, 1.0:1, 1.1:1, 1.2:1, 1.3:1, 1.4:1, 1.5:1, 2.0:1, 3.0:1, 4.0:1, 5.0:1, 6.0:1, 7.0 : 1, 8.0: 1, 9.0: 1 or 10.0: 1).
  • the reaction time after addition of the anhydride is from 0.1 to 24 hours (specifically 0.1, 1, 2, 3, 4, 5, 10, 15, 20 or 24 hours).
  • the separating and purifying step comprises the step of separating and purifying using ion exchange chromatography.
  • the P a and P b are independently selected from the group consisting of polyethylene glycol, polypropylene glycol, polyvinyl alcohol, polytetrahydrofuran, polypropylene oxide, polybutylene oxide, polyoxygen A residue of a copolymer of one or more of a heterocyclic butane and a polypropylene morpholine.
  • the P a and/or P b are polyethylene glycol residues.
  • the P a is a polyethylene glycol residue having a structure of R a —O—(CH 2 CH 2 O) m —, and R a is selected from the group consisting of: H, C 1- 6 alkyl, C 3-6 cycloalkyl and C 6-10 cycloalkyl, m is an integer from 170 to 565.
  • said R a is selected from: H, methyl, ethyl, isopropyl, cyclopropyl, cyclobutyl, cyclohexyl, and benzyl.
  • said R a is H or methyl.
  • the P a is a methoxypolyethylene glycol residue having the structure CH 3 O-(CH 2 CH 2 O) m -, and m is an integer from 170 to 565.
  • the P a may have a molecular weight of 7.5 to 25 KDa (specifically, 7.5, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19) , 20, 21, 22, 23, 24 or 25KDa).
  • the P b is a polyethylene glycol residue having a structure of R b —O—(CH 2 CH 2 O) n —, and R b is selected from the group consisting of: H, C 1- 6 alkyl, C 3-6 cycloalkyl and C 6-10 cycloalkyl, n being an integer from 170 to 565.
  • the R b is selected from the group consisting of H, methyl, ethyl, isopropyl, cyclopropyl, cyclobutyl, cyclohexyl and benzyl.
  • said R b is H or methyl.
  • the molecular weight of P b may be 7.5-25 KDa (specifically, 7.5, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 , 20, 21, 22, 23, 24 or 25KDa).
  • the m and n are equal integers.
  • the Y-branched hydrophilic polymeric carboxylic acid derivative is a Y-branched polyethylene glycol carboxylic acid derivative.
  • the X 1 is selected from the group consisting of: a single bond, -CH 2 -, -CH 2 CH 2 -, -CH 2 CH 2 CH 2 -, -CH 2 CH 2 CH 2 CH 2 - , -CH 2 CH 2 CH 2 CH 2 -, - CH (CH 3) -, - CH 2 CH (CH 3) -, - CH 2 CH 2 CH (CH 3) -, - CH 2 CH 2 CH (CH 3) -, - CH 2 CH 2 CH(CH 3 )-, -CH 2 CH 2 CH 2 CH 2 CH(CH 3 )-, -CH 2 CH 2 CH 2 CH 2 CH(CH 3 )-, -(CH 2 ) i O- And a combination of one or more of -(CH 2 ) i CO-, i is an integer from 0 to 5 (eg, 0, 1, 2, 3, 4 or 5).
  • said X 1 is -CH 2 CH 2 -.
  • the X 3 is selected from the group consisting of: a single bond, -CH 2 -, -CH 2 CH 2 -, -CH 2 CH 2 CH 2 -, -CH 2 CH 2 CH 2 CH 2 -, -CH 2 CH 2 CH 2 CH 2 -, - CH (CH 3) -, - CH 2 CH (CH 3) -, - CH 2 CH 2 CH (CH 3) -, - CH 2 CH 2 CH (CH 3) -, - CH 2 CH 2 CH(CH 3 )-, -CH 2 CH 2 CH 2 CH 2 CH(CH 3 )-, -CH 2 CH 2 CH 2 CH 2 CH(CH 3 )-, -(CH 2 ) i O- And a combination of one or more of -(CH 2 ) i CO-, i is an integer from 0 to 5 (eg, 0, 1, 2, 3, 4 or 5).
  • said X 3 is -CH 2 -.
  • the X 2 is
  • the R 3 is selected from the group consisting of: -H, -CH 3 , -CH 2 CH(CH 3 ) 2 , -CH(CH 3 )CH 2 CH 3 , -CH(CH 3 ) 2 , -CH 2 CH 2 SCH 3 -, -CH 2 OH, -CH 2 SH, -CH(OH)CH 3 , -CH 2 COOH and -CH 2 CH 2 COOH.
  • the X 2 is -CH 2 - or -CH(CH 3 )-.
  • the F is selected from the group consisting of: methoxy, ethoxy,
  • the F is
  • reaction of the preparation method is:
  • Another aspect of the present invention provides a Y-branched hydrophilic polymer carboxylic acid derivative prepared by the above method, which has the following structure:
  • P a , P b , X 1 , X 2 and X 3 have the above definitions of the present invention.
  • the carboxylic acid derivative has a molecular weight of 15-50 KDa (specifically 15, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48 or 50KDa).
  • the P a and P b are independently selected from the group consisting of polyethylene glycol, polypropylene glycol, polyvinyl alcohol, polytetrahydrofuran, polypropylene oxide, polybutylene oxide, polyoxygen A residue of a copolymer of one or more of a heterocyclic butane and a polypropylene morpholine.
  • the P a and/or P b are polyethylene glycol residues.
  • the P a is a polyethylene glycol residue having a structure of R a —O—(CH 2 CH 2 O) m —, and R a is selected from the group consisting of: H, C 1- 6 alkyl, C 3-6 cycloalkyl and C 6-10 cycloalkyl, m is an integer from 170 to 565.
  • said R a is selected from: H, methyl, ethyl, isopropyl, cyclopropyl, cyclobutyl, cyclohexyl, and benzyl.
  • said R a is H or methyl.
  • the P a is a methoxypolyethylene glycol residue having the structure CH 3 O-(CH 2 CH 2 O) m -, and m is an integer from 170 to 565.
  • the P a has a molecular weight of 7.5 to 25 KDa (specifically, 7.5, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24 or 25KDa).
  • the P b is a polyethylene glycol residue having a structure of R b —O—(CH 2 CH 2 O) n —, and R b is selected from the group consisting of: H, C 1- 6 alkyl, C 3-6 cycloalkyl and C 6-10 cycloalkyl, n being an integer from 170 to 565.
  • the R b is selected from the group consisting of H, methyl, ethyl, isopropyl, cyclopropyl, cyclobutyl, cyclohexyl and benzyl.
  • said R b is H or methyl.
  • the P b is a methoxypolyethylene glycol residue having the structure CH 3 O-(CH 2 CH 2 O) n -, n being an integer from 170 to 565.
  • the molecular weight of P b may be 7.5-25 KDa (specifically, 7.5, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 , 20, 21, 22, 23, 24 or 25KDa).
  • the Y-branched hydrophilic polymer carboxylic acid derivative is a Y-branched polyethylene glycol carboxylic acid derivative having the following structure:
  • the X 1 is selected from the group consisting of: a single bond, -CH 2 -, -CH 2 CH 2 -, -CH 2 CH 2 CH 2 -, -CH 2 CH 2 CH 2 CH 2 -, -CH 2 CH 2 CH 2 CH 2 -, -CH(CH 3 )-, -CH 2 CH(CH 3 )-, -CH 2 CH 2 CH(CH 3 )-, -CH 2 CH 2 CH 2 CH(CH 3 )-, -CH 2 CH 2 CH 2 CH(CH 3 )-, -CH 2 CH 2 CH 2 CH(CH 3 )-, -CH 2 CH 2 CH 2 CH 2 CH(CH 3 )-, -(CH 2 ) i O- and - i CO- combination of one or more of (2 CH), i is an integer (e.g., 3, 4 or 5) 0-5.
  • said X 1 is -CH 2 CH 2 -.
  • the X 3 is selected from the group consisting of: a single bond, -CH 2 -, -CH 2 CH 2 -, -CH 2 CH 2 CH 2 -, -CH 2 CH 2 CH 2 CH 2 - , -CH 2 CH 2 CH 2 CH 2 -, -CH(CH 3 )-, -CH 2 CH(CH 3 )-, -CH 2 CH 2 CH(CH 3 )-, -CH 2 CH 2 CH 2 CH(CH 3 )-, -CH 2 CH 2 CH 2 CH(CH 3 )-, -CH 2 CH 2 CH 2 CH(CH 3 )-, -CH 2 CH 2 CH 2 CH 2 CH(CH 3 )-, -(CH 2 ) i O- And a combination of one or more of -(CH 2 ) i CO-, i is an integer from 0 to 5 (eg, 0, 1, 2, 3, 4 or 5).
  • said X 3 is -CH 2 -.
  • the X 2 is
  • the carboxylic acid derivative has the structure:
  • the R 3 is selected from the group consisting of: -H, -CH 3 , -CH 2 CH(CH 3 ) 2 , -CH(CH 3 )CH 2 CH 3 , -CH(CH 3 ) 2 , -CH 2 CH 2 SCH 3 -, -CH 2 OH, -CH 2 SH, -CH(OH)CH 3 , -CH 2 COOH and -CH 2 CH 2 COOH.
  • said R 3 is -H or -CH 3 .
  • the m and n are equal integers.
  • Another aspect of the present invention also provides a Y-branched hydrophilic polymer derivative derived from the above carboxylic acid, which has the following structure:
  • X 4 is a linking group selected from the group consisting of: -(CH 2 ) j -, -(CH 2 ) j O-, -(CH 2 ) j S-, -(CH 2 ) j CO-, -(CH 2 ) a combination of one or more of j NH-, -(CH 2 ) j CONH- and -(CH 2 ) j NHCO-, j is an integer from 0 to 10,
  • Q is a terminal group selected from: C 1-6 alkoxy, hydroxy, amino, carboxy, thiol, ester, keto, aldehyde, o-dithiopyridyl, azide, hydrazide, alkynyl , silyl, maleimide and succinimide groups.
  • the derivative may have a molecular weight of 15-50 KDa (specifically 15, 16, 16, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48 or 50 KDa).
  • the P a and P b are independently selected from the group consisting of polyethylene glycol, polypropylene glycol, polyvinyl alcohol, polytetrahydrofuran, polypropylene oxide, polybutylene oxide, polyoxygen A residue of a copolymer of one or more of a heterocyclic butane and a polypropylene morpholine.
  • the P a and/or P b are polyethylene glycol residues.
  • the P a is a polyethylene glycol residue having a structure of R a —O—(CH 2 CH 2 O) m —, and R a is selected from the group consisting of: H, C 1- 6 alkyl, C 3-6 cycloalkyl and C 6-10 cycloalkyl, m is an integer from 170 to 565.
  • said R a is selected from: H, methyl, ethyl, isopropyl, cyclopropyl, cyclobutyl, cyclohexyl, and benzyl.
  • said R a is H or methyl.
  • the P a is a methoxypolyethylene glycol residue having the structure CH 3 O-(CH 2 CH 2 O) m -, and m is an integer from 170 to 565.
  • the P a has a molecular weight of 7.5 to 25 KDa (specifically, 7.5, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24 or 25KDa).
  • the P b is a polyethylene glycol residue having a structure of R b —O—(CH 2 CH 2 O) n —, and R b is selected from the group consisting of: H, C 1- 6 alkyl, C 3-6 cycloalkyl and C 6-10 cycloalkyl, n being an integer from 170 to 565.
  • the R b is selected from the group consisting of H, methyl, ethyl, isopropyl, cyclopropyl, cyclobutyl, cyclohexyl and benzyl.
  • said R b is H or methyl.
  • the molecular weight of P b may be 7.5-25 KDa (specifically, 7.5, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 , 20, 21, 22, 23, 24 or 25KDa).
  • the Y-branched hydrophilic polymer derivative is a Y-branched polyethylene glycol derivative having the following structure:
  • the X 4 is selected from: a single bond, -CH 2 -, - CH 2 CH 2 -, - CH 2 CH 2 CH 2 -, - CH 2 CH 2 CH 2 CH 2 -, -CH 2 CH 2 CH 2 CH 2 -, -(CH 2 ) j CO-, -(CH 2 ) j NH-, -(CH 2 ) j CONH- and -(CH 2 ) j NHCO-
  • j is an integer from 0 to 5 (eg, 0, 1, 2, 3, 4, or 5).
  • the X 4 is selected from the group consisting of: a single bond, -CH 2 -, -CH 2 CH 2 -, -CH 2 CH 2 CH 2 -, -CH 2 NH-, -CH 2 CH 2 NH -, - CH 2 CH 2 CH 2 NH -, - CH 2 CONH -, - CH 2 CH 2 CONH- , and combinations -CH 2 CH 2 CH 2 CONH- of one or more.
  • the Q is selected from the group consisting of: -OH, -SH, -NH 2 , -COOH, -CHO, And -N 3 .
  • the Y-branched hydrophilic polymer derivative has the following structure:
  • the R 3 is selected from the group consisting of: -H, -CH 3 , -CH 2 CH(CH 3 ) 2 , -CH(CH 3 )CH 2 CH 3 , -CH(CH 3 ) 2 , -CH 2 CH 2 SCH 3 -, -CH 2 OH, -CH 2 SH, -CH(OH)CH 3 , -CH 2 COOH and -CH 2 CH 2 COOH.
  • said R 3 is -H or -CH 3 .
  • the m and n are equal integers.
  • Another aspect of the present invention provides a process for producing the above Y-branched hydrophilic polymer derivative, which comprises the steps of the preparation method of the above Y-branched hydrophilic polymer carboxylic acid derivative.
  • Another aspect of the present invention provides a use of the above Y-branched hydrophilic polymer carboxylic acid derivative, Y-branched hydrophilic polymer derivative in a modified drug.
  • Another aspect of the present invention provides a combination of the above Y-branched hydrophilic polymer carboxylic acid derivative, Y-branched hydrophilic polymer derivative and a drug of the present invention.
  • the drug is selected from the group consisting of amino acids, polypeptides, proteins, sugars, organic acids, alkaloids, flavonoids, terpenoids, terpenoids, phenylpropanoid phenols, steroids, and steroids. drug.
  • Another aspect of the present invention provides a method for producing the above-mentioned Y-branched hydrophilic polymer carboxylic acid derivative of the present invention, in the pharmaceutical composition for preparing the above-mentioned Y-branched hydrophilic polymer carboxylic acid derivative application.
  • Another aspect of the present invention provides a method for preparing the above-mentioned Y-branched hydrophilic polymer carboxylic acid derivative of the present invention, in the preparation of the above Y-branched hydrophilic polymer derivative, and a pharmaceutical combination thereof .
  • Another aspect of the invention also provides a pharmaceutical composition
  • a pharmaceutical composition comprising the above combination of the invention and, optionally, a pharmaceutically acceptable carrier or excipient.
  • Another aspect of the present invention provides a Y-branched hydrophilic polymer carboxylic acid derivative, a Y-branched polyethylene glycol derivative, and a pharmaceutical conjugate thereof and a pharmaceutical composition thereof, in the preparation of a therapeutic disease The application of the drug.
  • the preparation method of the Y-branched hydrophilic polymer carboxylic acid derivative (especially the high-purity high molecular weight Y-branched polyethylene glycol carboxylic acid derivative) provided by the invention has simple preparation steps but the product after the reaction The separation is easy, the separation cost is low, the product purity and the yield are high, and the preparation of other derivatives based on the preparation of the carboxylic acid derivative and the drug combination thereof is facilitated, which is advantageous for industrial amplification and commercial application.
  • the prepared Y-branched hydrophilic polymer carboxylic acid derivative (especially the high molecular weight Y-branched polyethylene glycol carboxylic acid derivative) has high purity and high commercial value, especially in the preparation of prevention and/or Or the application of drugs for the treatment of diseases.
  • Figure 1 is a GFC chromatogram of the crude product before column separation provided in Example 1 of the present invention.
  • Example 2 is a chromatogram of a collection starting point GFC provided in Example 1 of the present invention.
  • Fig. 3 is a view showing a peak point GFC chromatogram provided in Example 1 of the present invention.
  • Example 4 is a GFC chromatogram of the post-column product provided in Example 1 of the present invention.
  • Fig. 5 is a GFC chromatogram of the crude product before column separation according to Example 2 of the present invention.
  • Figure 6 is a chromatogram of the collection starting point GFC provided in Example 2 of the present invention.
  • Fig. 7 is a view showing a peak point GFC chromatogram provided in Example 2 of the present invention.
  • Figure 8 is a GFC chromatogram of the post-column product provided in Example 2 of the present invention.
  • Figure 9 is a GFC chromatogram of the pre-column crude product provided in Example 3 of the present invention.
  • Figure 10 is a chromatogram of the collection starting point GFC provided in Example 3 of the present invention.
  • Figure 11 is a view showing a peak point GFC chromatogram provided in Example 3 of the present invention.
  • Figure 12 is a GFC chromatogram of the post-column product provided in Example 3 of the present invention.
  • Figure 13 is a GFC chromatogram of the crude product before column separation according to Example 4 of the present invention.
  • Figure 14 is a chromatogram of the collection starting point GFC provided in Example 4 of the present invention.
  • Figure 15 is a view showing a peak point GFC chromatogram provided in Example 4 of the present invention.
  • Figure 16 is a GFC chromatogram of the post-column product provided in Example 4 of the present invention.
  • Figure 17 is a GFC chromatogram of the pre-column crude product provided in Example 5 of the present invention.
  • Figure 18 is a chromatogram of the collection starting point GFC provided in Example 5 of the present invention.
  • Fig. 19 is a view showing a peak point GFC chromatogram provided in Example 5 of the present invention.
  • Figure 20 is a GFC chromatogram of the post-column product provided in Example 5 of the present invention.
  • Alkyl refers to a hydrocarbon chain radical that is linear or branched and free of unsaturated bonds, and which is attached to the rest of the molecule by a single bond.
  • the C1-C6 alkyl group means an alkyl group having 1 to 6 carbon atoms, such as methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, t-butyl, n-pentyl, iso Amyl, neopentyl, tert-amyl, n-hexyl, isohexyl and the like.
  • alkyl group is substituted by a cycloalkyl group, it is correspondingly a "cycloalkylalkyl” radical such as cyclopropylmethyl, cyclopropylethyl, cyclobutylmethyl, cyclopentylmethyl, cyclohexylmethyl, etc. .
  • cycloalkylalkyl such as cyclopropylmethyl, cyclopropylethyl, cyclobutylmethyl, cyclopentylmethyl, cyclohexylmethyl, etc.
  • an aryl group it is correspondingly an "aralkyl” radical such as benzyl, benzhydryl or phenethyl.
  • heterocyclic group it is correspondingly a "heterocyclylalkyl” radical.
  • alkoxy means a substituent formed by substituting a hydrogen in a hydroxy group with an alkyl group
  • alkoxy group of C1-C6 means an alkoxy group having 1 to 6 carbon atoms, such as a methoxy group or an ethoxy group. , propoxy, butoxy, and the like.
  • Cycloalkyl means an alicyclic hydrocarbon such as containing from 1 to 4 monocyclic and/or fused rings containing from 3 to 18 carbon atoms, preferably from 3 to 10 carbon atoms, such as cyclopropyl, cyclohexyl or Adamantyl and the like
  • the C3-C6 cycloalkyl group in the present invention means a cycloalkyl group having 3 to 6 carbon atoms such as a cyclopropyl group, a cyclobutyl group, a cyclopentyl group and a cyclohexyl group.
  • a “substituted” group as used in the present invention refers to a group substituted at one or more of the available sites by one or more suitable groups, specifically, for example, a substituted alkyl group, which refers to an alkyl group.
  • One or more hydrogens are substituted by one or more suitable groups such as an alkyl group (e.g., a C1-6 alkyl group, particularly a C1-3 alkyl group such as a methyl group, an ethyl group).
  • alkoxy such as C1-6 alkoxy, especially C1-3 alkoxy, such as methoxy, ethoxy or propoxy
  • alkenyl such as An alkenyl group of C1-6, especially an alkenyl group of C1-3, such as a vinyl group, an alkynyl group (such as an alkynyl group of C1-6, especially an alkynyl group of C1-3, such as a propynyl group), a cycloalkyl group (such as a cycloalkyl group of C3-6, such as cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl), an aryl group (such as an aryl group of C6-12, especially a phenyl group), an aryloxy group (such as benzene) An oxy), an alkylaryl group (such as a benzyl group), a heterocyclic group (such as a heterocycl
  • some specific groups and chemical structures involved in the present invention correspond to the following: hydroxyl group, -OH; amino group, -NH 2 ; carboxyl group, Sulfhydryl, -SH; ester group, (wherein Q 1 may be an alkyl group, an aryl group or a heterocyclic group such as methyl, ethyl, n-propyl, t-butyl, maleimide, succinimidyl, Ketone, (wherein Q 2 may be substituted or unsubstituted alkyl, aryl, heterocyclic, such as substituted or unsubstituted methyl, ethyl, n-propyl, Ethyl, -CHO; o-dithiopyridyl, Azido group, Acyl hydrazino, Alkynyl, Silyl group, (wherein Q 3 may be the same or different alkyl or alkoxy group, such as methyl, ethyl, propyl
  • the definition of the linking group refers to the group two linking groups listed in the above chemical bond formed by the engagement of the connection, for example - (CH 2) j - and -
  • the combination of (CH 2 ) j NHCO- may be -(CH 2 ) j NHCO(CH 2 ) j -; specifically, the combination of -CH 2 - and -CH 2 CH 2 NHCO- may be -CH 2 CH 2 NHCOCH 2 -, -CH 2 CH 2 CH 2 NHCO-.
  • the "combination” is used to define the chemical structure of the linking group, and does not involve the preparation steps, combination order, and the like of the linking group.
  • the Y-branched polyethylene glycol-acetic acid having a molecular weight of 20,000 was prepared by the synthesis method of Example 5 of Patent CN1243779C: 10 g of polyethylene glycol monomethyl ether-aminoacetic acid (mPEG-Gly) having a molecular weight of 10,000 and a molecular weight of 10 g 10000 of polyethylene glycol monomethyl ether-carboxyacetate succinimide ester (mPEG-OCH 2 CO-NHS) was dissolved in 200 ml of dichloromethane, 0.11 ml of triethylamine was added to the solution, and the reaction was carried out overnight at room temperature.
  • mPEG-Gly polyethylene glycol monomethyl ether-aminoacetic acid
  • mPEG-OCH 2 CO-NHS polyethylene glycol monomethyl ether-carboxyacetate succinimide ester
  • the solvent was concentrated, the residue was added diethyl ether, and the precipitate was collected by filtration, dried in vacuo, purified by ion-exchange chromatography column, and monitored by GFC, and collected at a peak height of the target product exceeding 5 mv, and collected at less than 5 mv.
  • Step of reacting mPEG-Gly with mPEG-OCH 2 CO-NHS Referring to Example 2, after reacting at room temperature overnight, BOC anhydride was added, the reaction was carried out for 3 h, the solvent was concentrated by rotary evaporation, the residue was added diethyl ether, and the precipitate was collected by filtration, dried under vacuum, and ion exchange Column purification, GFC monitoring, starting with the peak height of the target product exceeding 5 mv, and collecting at less than 5 mv.
  • the Y-branched polyethylene glycol-acetic acid having a molecular weight of 44,000 was prepared by the synthesis method of Example 5 of Patent CN1243779C: 10g of polyethylene glycol monomethyl ether-aminoacetic acid (mPEG-Gly) having a molecular weight of 22000 and a molecular weight of 10g 22000 of polyethylene glycol monomethyl ether-carboxyacetate succinimide ester (mPEG-OCH 2 CO-NHS) was dissolved in 200 ml of dichloromethane, 0.23 ml of triethylamine was added to the solution, and the reaction was carried out overnight at room temperature.
  • mPEG-Gly polyethylene glycol monomethyl ether-aminoacetic acid
  • mPEG-OCH 2 CO-NHS polyethylene glycol monomethyl ether-carboxyacetate succinimide ester
  • the solvent was concentrated, the residue was added diethyl ether, and the precipitate was collected by filtration, dried in vacuo, purified by ion-exchange chromatography column, and monitored by GFC, and collected at a peak height of the target product exceeding 5 mv, and collected at less than 5 mv.
  • Step of reacting mPEG-Gly with mPEG-OCH 2 CO-NHS Referring to Example 4, after reacting at room temperature overnight, adding BOC anhydride, reacting for 3 hours, concentrating the solvent by rotary evaporation, adding the residue to diethyl ether, collecting the precipitate by filtration, vacuum drying, ion exchange Column purification, GFC monitoring, starting with the peak height of the target product exceeding 5 mv, and collecting at less than 5 mv.
  • the effect of the prior art on product purification will gradually increase with increasing molecular weight, and the method of the present invention can significantly improve purification yield and product purity for high molecular weight products.

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Abstract

Disclosed in the present invention is a method for preparing a Y-branched hydrophilic polymer carboxylic acid derivative, in particular a method for preparing a Y-branched polyethylene glycol carboxylic acid derivative having a high purity and high molecular weight. The preparation steps are simple, the product after reaction is easy to be separated, the cost for separation is low, the purity and yield of the product are high, facilitating the subsequent preparation of other derivatives and medicament conjugates based on the preparation of the carboxylic acid derivative, being advantageous for industrial amplification and commercial applications. The prepared Y-branched hydrophilic polymer carboxylic acid derivative (in particular the Y-branched polyethylene glycol carboxylic acid derivative having a high molecular weight) product has a high purity and high commercial application value, in particular has use in the preparation of medicaments for preventing and/or treating diseases.

Description

一种Y型分支的亲水性聚合物羧酸衍生物的制备方法Method for preparing Y-branched hydrophilic polymer carboxylic acid derivative 技术领域Technical field
本发明涉及聚合物技术领域，具体涉及一种Y型分支的亲水性聚合物羧酸衍生物的制备方法，特别是一种高纯度的高分子量Y型分支聚乙二醇羧酸衍生物的制备方法。The invention relates to the technical field of polymers, in particular to a method for preparing a Y-branched hydrophilic polymer carboxylic acid derivative, in particular to a high-purity high molecular weight Y-branched polyethylene glycol carboxylic acid derivative. Preparation.
背景技术Background technique
聚乙二醇(PEG)被认为是已知聚合物中蛋白和细胞吸收水平非常低的聚合物，而且，其具有良好的水溶性和生物相容性、无毒、无免疫性、无致畸性且无抗原性的优势，被广泛应用于药物修饰、制剂制备与医疗器械材料等领域。从1991年开始，第一种用PEG修饰的药物PEG-ADA被FDA批准上市后，各大制药公司对PEG在药物领域的研发投入了极大的精力和金钱。近几年来，上市的产品有PEG-生长抑素、PEG-干扰素、PEG-粒细胞集落因子等。目前，尚有几十种PEG修饰药物处于研究或临床试验阶段。Polyethylene glycol (PEG) is considered to be a polymer with a very low level of protein and cellular absorption in known polymers, and it has good water solubility and biocompatibility, non-toxicity, non-immunity, and no teratogenicity. The advantages of sex and no antigenicity are widely used in the fields of drug modification, preparation preparation and medical device materials. Since 1991, after the first PEG-modified drug PEG-ADA was approved by the FDA, major pharmaceutical companies have invested a lot of energy and money in the research and development of PEG in the pharmaceutical field. In recent years, the products listed have PEG-somatostatin, PEG-interferon, PEG-granulocyte colony factor and the like. At present, there are dozens of PEG-modified drugs in the research or clinical trial stage.
在药物修饰领域，Y型聚乙二醇是一类被广泛使用的聚乙二醇，其可明显降低修饰后药物的活性损失，特别是在改善蛋白质及多肽药物的临床应用方面，能更有效地阻止抗体接近蛋白质药物，从而大大增加蛋白质药物在体内的循环半衰期，同时大大降低其在体内的免疫源性。其中，Y型PEG的端基为羧基时，可与修饰药物或其他化合物上的氨基、羟基、硫羟基等反应形成共价键实现修饰连接，是一种常用的Y型PEG衍生物。其制备方法如专利CN1243779C中所述，
Figure PCTCN2018077229-appb-000001
可通过
Figure PCTCN2018077229-appb-000002
与
Figure PCTCN2018077229-appb-000003
反应得到，反应结束后经离子交换色谱纯化。在阴离子交换柱上，化合物中所含-COOH基团数越多，在柱上的吸附力越强。因此，当PEG结构中-COOH个数相同时，分子量越低，在柱上的吸附力越强。并且，这种分子量带来的吸附力的差距，会随着分子量的递增而减弱。但是，由于mPEG-gly中-NH-的存在会抵消部分-COOH的酸性，从而降低了该杂质在离子交换柱上的吸附力，以上三个化合物在柱上的吸附力强弱排序为mPEG-cm、mPEG-gly、Y-cm。上述制备反应产物中包括目标产物Y-cm和未反应的反应物mPEG-gly和mPEG-cm，在高分子量的范围内，mPEG-gly会对分离产生很大的干扰，使目标产物的纯度和收率降低。如CN1243779C的制备实施例中Y型PEG产物的产率较低，仅为50％，产品分离困难，成本较高，不利于工业放大。 In the field of drug modification, Y-type polyethylene glycol is a widely used polyethylene glycol, which can significantly reduce the loss of activity of modified drugs, especially in improving the clinical application of protein and peptide drugs. It prevents the antibody from approaching the protein drug, thereby greatly increasing the circulating half-life of the protein drug in the body while greatly reducing its immunogenicity in vivo. Wherein, when the terminal group of the Y-type PEG is a carboxyl group, it can react with an amino group, a hydroxyl group or a thiol group on a modified drug or other compound to form a covalent bond to realize a modified linkage, and is a commonly used Y-type PEG derivative. The preparation method thereof is as described in the patent CN1243779C.
Figure PCTCN2018077229-appb-000001
accessible
Figure PCTCN2018077229-appb-000002
versus
Figure PCTCN2018077229-appb-000003
The reaction was obtained, and after completion of the reaction, it was purified by ion-exchange chromatography. On the anion exchange column, the more the number of -COOH groups contained in the compound, the stronger the adsorption force on the column. Therefore, when the number of -COOH in the PEG structure is the same, the lower the molecular weight, the stronger the adsorption force on the column. Moreover, the difference in adsorption force due to this molecular weight decreases as the molecular weight increases. However, since the presence of -NH- in mPEG-gly counteracts the acidity of the moiety -COOH, thereby reducing the adsorption of the impurity on the ion exchange column, the adsorption strength of the above three compounds on the column is ranked as mPEG- Cm, mPEG-gly, Y-cm. The above preparation reaction product includes the target product Y-cm and the unreacted reactants mPEG-gly and mPEG-cm. In the range of high molecular weight, mPEG-gly greatly interferes with the separation, so that the purity of the target product is The yield is lowered. For example, in the preparation example of CN1243779C, the yield of the Y-type PEG product is low, only 50%, the product separation is difficult, and the cost is high, which is not favorable for industrial amplification.
发明内容Summary of the invention
为克服现有技术的不足，本发明提供了一种Y型分支的亲水性聚合物羧酸衍生物的制备方法。To overcome the deficiencies of the prior art, the present invention provides a process for the preparation of a Y-branched hydrophilic polymeric carboxylic acid derivative.
所述方法包括如下反应：The method includes the following reaction:
Figure PCTCN2018077229-appb-000004
Figure PCTCN2018077229-appb-000004
其中，P a和P b为相同或不同的亲水性聚合物残基， Wherein P a and P b are the same or different hydrophilic polymer residues,
X 1和X 3为连接基团，独立地选自：-(CH 2) i-、
Figure PCTCN2018077229-appb-000005
-(CH 2) iO-、-(CH 2) iS-和-(CH 2) iCO-中的一种或多种的组合，i为0-10的整数， X 1 and X 3 are a linking group independently selected from: -(CH 2 ) i -,
Figure PCTCN2018077229-appb-000005
a combination of one or more of -(CH 2 ) i O-, -(CH 2 ) i S- and -(CH 2 ) i CO-, i is an integer from 0 to 10,
X 2为连接基团，选自：-(CH 2) r-、-(CH 2) rO-、-(CH 2) rS-和
Figure PCTCN2018077229-appb-000006
中的一种或多种的组合，r为0-10的整数， X 2 is a linking group, chosen from :-( CH 2) r -, - (CH 2) r O -, - (CH 2) r S- , and
Figure PCTCN2018077229-appb-000006
a combination of one or more of the following, r is an integer from 0 to 10,
F为端基，选自：C 1-6的取代或未取代的烷基、C 1-6的取代或未取代的烷氧基、
Figure PCTCN2018077229-appb-000007
F is a terminal group selected from: substituted or unsubstituted C 1-6 alkyl, C 1-6 substituted or unsubstituted alkoxy group,
Figure PCTCN2018077229-appb-000007
Figure PCTCN2018077229-appb-000008
Figure PCTCN2018077229-appb-000008
R 1和R 2独立地选自：-H、C 1-6取代或未取代的烷基、C 1-6取代或未取代的烷氧基、C 3-6取代或未取代的环烷基和C 4-10取代或未取代的亚烷基环烷基， R 1 and R 2 are independently selected from: -H, C 1-6 substituted or unsubstituted alkyl, C 1-6 substituted or unsubstituted alkoxy, C 3-6 substituted or unsubstituted cycloalkyl And a C 4-10 substituted or unsubstituted alkylene cycloalkyl group,
R 3选自：-H、C 1-6取代或未取代的烷基、C 6-10取代或未取代的芳烷基和C 4-10取代或未取代的杂环烷基， R 3 is selected from the group consisting of: -H, a C 1-6 substituted or unsubstituted alkyl group, a C 6-10 substituted or unsubstituted aralkyl group, and a C 4-10 substituted or unsubstituted heterocycloalkyl group,
反应结束后，加入酸酐，继续反应，分离纯化。After completion of the reaction, an acid anhydride is added, and the reaction is continued and separated and purified.
上述制备方法更适于制备较高分子量的Y型分支的亲水性聚合物羧酸衍生物，反应收率和产品纯度更高，分离更容易。在本发明的一个实施例中，所述Y型分支的亲水性聚合物羧酸衍生物的分子量可为15-50KDa(具体可为15、16、18、20、22、24、26、28、30、32、34、36、38、40、42、44、46、48或50KDa)。The above preparation method is more suitable for preparing a higher molecular weight Y-branched hydrophilic polymer carboxylic acid derivative, and the reaction yield and product purity are higher, and separation is easier. In one embodiment of the present invention, the Y-branched hydrophilic polymer carboxylic acid derivative may have a molecular weight of 15 to 50 KDa (specifically 15, 16, 16, 20, 22, 24, 26, 28) , 30, 32, 34, 36, 38, 40, 42, 44, 46, 48 or 50 KDa).
在本发明的一个实施例中，所述酸酐为有机酸酸酐。In one embodiment of the invention, the anhydride is an organic acid anhydride.
在本发明的一个实施例中，所述有机酸酸酐选自：二碳酸二叔丁酯(Boc酸酐)、乙酸酐、丙酸酐、异丁酸酐、丁酸酐、苯甲酸酐和邻苯二甲酸酐中的一种或多种。In one embodiment of the invention, the organic acid anhydride is selected from the group consisting of: di-tert-butyl dicarbonate (Boc anhydride), acetic anhydride, propionic anhydride, isobutyric anhydride, butyric anhydride, benzoic anhydride, and phthalic anhydride. One or more of them.
在本发明的一个优选实施例中，所述酸酐为Boc酸酐。In a preferred embodiment of the invention, the anhydride is a Boc anhydride.
在本发明的一个实施例中，所述酸酐与反应物
Figure PCTCN2018077229-appb-000009
加入量的摩尔比为0.01-10:1(具体可为0.01:1、0.1:1、0.2:1、0.3:1、0.4:1、0.5:1、0.6:1、0.7:1、0.8:1、0.9:1、1.0:1、1.1:1、1.2:1、1.3:1、1.4:1、1.5:1、2.0:1、3.0:1、4.0:1、5.0:1、6.0:1、7.0:1、8.0:1、9.0:1或10.0:1)。 In one embodiment of the invention, the anhydride and reactant
Figure PCTCN2018077229-appb-000009
The molar ratio of the added amount is 0.01-10:1 (specifically, 0.01:1, 0.1:1, 0.2:1, 0.3:1, 0.4:1, 0.5:1, 0.6:1, 0.7:1, 0.8:1) , 0.9:1, 1.0:1, 1.1:1, 1.2:1, 1.3:1, 1.4:1, 1.5:1, 2.0:1, 3.0:1, 4.0:1, 5.0:1, 6.0:1, 7.0 : 1, 8.0: 1, 9.0: 1 or 10.0: 1).
在本发明的一个实施例中，加入酸酐后反应时间为0.1-24小时(具体可为0.1、1、2、3、4、5、10、15、20或24小时)。In one embodiment of the invention, the reaction time after addition of the anhydride is from 0.1 to 24 hours (specifically 0.1, 1, 2, 3, 4, 5, 10, 15, 20 or 24 hours).
在本发明的一个实施例中，所述分离纯化步骤包括采用离子交换色谱分离纯化的步骤。In one embodiment of the invention, the separating and purifying step comprises the step of separating and purifying using ion exchange chromatography.
在本发明的一个实施例中，所述的P a和P b独立地选自：聚乙二醇、聚丙二醇、聚乙烯醇、聚四氢呋喃、聚环氧丙烷、聚环氧丁烷、聚氧杂环丁烷和聚丙烯吗啉中的一种或多种的共聚物的残基。 In one embodiment of the invention, the P a and P b are independently selected from the group consisting of polyethylene glycol, polypropylene glycol, polyvinyl alcohol, polytetrahydrofuran, polypropylene oxide, polybutylene oxide, polyoxygen A residue of a copolymer of one or more of a heterocyclic butane and a polypropylene morpholine.
在本发明的一个优选实施例中，所述的P a和/或P b为聚乙二醇残基。 In a preferred embodiment of the invention, the P a and/or P b are polyethylene glycol residues.
在本发明的一个实施例中，所述的P a为聚乙二醇残基，其结构为R a-O-(CH 2CH 2O) m-，R a选自：H、C 1-6烷基、C 3-6环烷基和C 6-10环烷基，m为170-565的整数。 In one embodiment of the present invention, the P a is a polyethylene glycol residue having a structure of R a —O—(CH 2 CH 2 O) m —, and R a is selected from the group consisting of: H, C 1- 6 alkyl, C 3-6 cycloalkyl and C 6-10 cycloalkyl, m is an integer from 170 to 565.
在本发明的一个实施例中，所述R a选自：H、甲基、乙基、异丙基、环丙基、环丁基、环己基和苄基。 In one embodiment of the present invention, said R a is selected from: H, methyl, ethyl, isopropyl, cyclopropyl, cyclobutyl, cyclohexyl, and benzyl.
在本发明的一个优选实施例中，所述R a为H或甲基。 In a preferred embodiment of the invention, said R a is H or methyl.
在本发明的一个实施例中，所述的P a为甲氧基聚乙二醇残基，其结构为CH 3O-(CH 2CH 2O) m-，m为170-565的整数。 In one embodiment of the invention, the P a is a methoxypolyethylene glycol residue having the structure CH 3 O-(CH 2 CH 2 O) m -, and m is an integer from 170 to 565.
在本发明的一个实施例中，所述的P a的分子量可为7.5-25KDa(具体可为7.5、8、9、10、11、12、13、14、15、16、17、18、19、20、21、22、23、24或25KDa)。 In one embodiment of the present invention, the P a may have a molecular weight of 7.5 to 25 KDa (specifically, 7.5, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19) , 20, 21, 22, 23, 24 or 25KDa).
在本发明的一个实施例中，所述的P b为聚乙二醇残基，其结构为R b-O-(CH 2CH 2O) n-，R b选自：H、C 1-6烷基、C 3-6环烷基和C 6-10环烷基，n为170-565的整数。 In one embodiment of the present invention, the P b is a polyethylene glycol residue having a structure of R b —O—(CH 2 CH 2 O) n —, and R b is selected from the group consisting of: H, C 1- 6 alkyl, C 3-6 cycloalkyl and C 6-10 cycloalkyl, n being an integer from 170 to 565.
在本发明的一个实施例中，所述R b选自：H、甲基、乙基、异丙基、环丙基、环丁基、环己基和苄基。 In one embodiment of the invention, the R b is selected from the group consisting of H, methyl, ethyl, isopropyl, cyclopropyl, cyclobutyl, cyclohexyl and benzyl.
在本发明的一个优选实施例中，所述R b为H或甲基。 In a preferred embodiment of the invention, said R b is H or methyl.
在本发明的一个实施例中，所述的P b为甲氧基聚乙二醇残基，其结构为CH 3O-(CH 2CH 2O) n-，n为170-565的整数。 In one embodiment of the present invention, the P b using methoxy polyethylene glycol residue having the structure CH 3 O- (CH 2 CH 2 O) n -, n is an integer of 170-565.
在本发明的一个实施例中，所述的P b的分子量可为7.5-25KDa(具体可为7.5、8、9、10、11、12、13、14、15、16、17、18、19、20、21、22、23、24或25KDa)。 In one embodiment of the present invention, the molecular weight of P b may be 7.5-25 KDa (specifically, 7.5, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 , 20, 21, 22, 23, 24 or 25KDa).
在本发明的一个实施例中，所述m与n为相等的整数。In one embodiment of the invention, the m and n are equal integers.
在本发明的一个优选实施例中，所述Y型分支的亲水性聚合物羧酸衍生物为Y型分支的聚乙二醇羧酸衍生物所述反应如下：In a preferred embodiment of the invention, the Y-branched hydrophilic polymeric carboxylic acid derivative is a Y-branched polyethylene glycol carboxylic acid derivative. The reaction is as follows:
Figure PCTCN2018077229-appb-000010
Figure PCTCN2018077229-appb-000010
在本发明的一个实施例中，所述X 1选自：单键、-CH 2-、-CH 2CH 2-、-CH 2CH 2CH 2-、-CH 2CH 2CH 2CH 2-、-CH 2CH 2CH 2CH 2CH 2-、-CH(CH 3)-、-CH 2CH(CH 3)-、-CH 2CH 2CH(CH 3)-、-CH 2CH 2CH 2CH(CH 3)-、-CH 2CH 2CH 2CH 2CH(CH 3)-、-CH 2CH 2CH 2CH 2CH 2CH(CH 3)-、-(CH 2) iO-和-(CH 2) iCO-中的一种或多种的组合，i为0-5的整数(如0、1、2、3、4或5)。 In one embodiment of the invention, the X 1 is selected from the group consisting of: a single bond, -CH 2 -, -CH 2 CH 2 -, -CH 2 CH 2 CH 2 -, -CH 2 CH 2 CH 2 CH 2 - , -CH 2 CH 2 CH 2 CH 2 CH 2 -, - CH (CH 3) -, - CH 2 CH (CH 3) -, - CH 2 CH 2 CH (CH 3) -, - CH 2 CH 2 CH 2 CH(CH 3 )-, -CH 2 CH 2 CH 2 CH 2 CH(CH 3 )-, -CH 2 CH 2 CH 2 CH 2 CH 2 CH(CH 3 )-, -(CH 2 ) i O- And a combination of one or more of -(CH 2 ) i CO-, i is an integer from 0 to 5 (eg, 0, 1, 2, 3, 4 or 5).
在本发明的一个优选实施例中，所述X 1为-CH 2CH 2-。 In a preferred embodiment of the invention, said X 1 is -CH 2 CH 2 -.
在本发明的一个实施例中，所述X 3选自：单键、-CH 2-、-CH 2CH 2-、-CH 2CH 2CH 2-、-CH 2CH 2CH 2CH 2-、-CH 2CH 2CH 2CH 2CH 2-、-CH(CH 3)-、-CH 2CH(CH 3)-、-CH 2CH 2CH(CH 3)-、-CH 2CH 2CH 2CH(CH 3)-、-CH 2CH 2CH 2CH 2CH(CH 3)-、-CH 2CH 2CH 2CH 2CH 2CH(CH 3)-、-(CH 2) iO-和-(CH 2) iCO-中的一种或多种的组合，i为0-5的整数(如0、1、2、3、4或5)。 In one embodiment of the invention, the X 3 is selected from the group consisting of: a single bond, -CH 2 -, -CH 2 CH 2 -, -CH 2 CH 2 CH 2 -, -CH 2 CH 2 CH 2 CH 2 - , -CH 2 CH 2 CH 2 CH 2 CH 2 -, - CH (CH 3) -, - CH 2 CH (CH 3) -, - CH 2 CH 2 CH (CH 3) -, - CH 2 CH 2 CH 2 CH(CH 3 )-, -CH 2 CH 2 CH 2 CH 2 CH(CH 3 )-, -CH 2 CH 2 CH 2 CH 2 CH 2 CH(CH 3 )-, -(CH 2 ) i O- And a combination of one or more of -(CH 2 ) i CO-, i is an integer from 0 to 5 (eg, 0, 1, 2, 3, 4 or 5).
在本发明的一个优选实施例中，所述X 3为-CH 2-。 In a preferred embodiment of the invention, said X 3 is -CH 2 -.
在本发明的一个实施例中，所述X 2为
Figure PCTCN2018077229-appb-000011
In an embodiment of the invention, the X 2 is
Figure PCTCN2018077229-appb-000011
在本发明的一个实施例中，所述R 3选自：-H、-CH 3、-CH 2CH(CH 3) 2、-CH(CH 3)CH 2CH 3、-CH(CH 3) 2、
Figure PCTCN2018077229-appb-000012
-CH 2CH 2SCH 3-、-CH 2OH、-CH 2SH、-CH(OH)CH 3、
Figure PCTCN2018077229-appb-000013
-CH 2COOH和-CH 2CH 2COOH。 In one embodiment of the invention, the R 3 is selected from the group consisting of: -H, -CH 3 , -CH 2 CH(CH 3 ) 2 , -CH(CH 3 )CH 2 CH 3 , -CH(CH 3 ) 2 ,
Figure PCTCN2018077229-appb-000012
-CH 2 CH 2 SCH 3 -, -CH 2 OH, -CH 2 SH, -CH(OH)CH 3 ,
Figure PCTCN2018077229-appb-000013
-CH 2 COOH and -CH 2 CH 2 COOH.
在本发明的一个优选实施例中，所述X 2为-CH 2-或-CH(CH 3)-。 In a preferred embodiment of the invention, the X 2 is -CH 2 - or -CH(CH 3 )-.
在本发明的一个实施例中，所述F选自：甲氧基、乙氧基、
Figure PCTCN2018077229-appb-000014
In one embodiment of the invention, the F is selected from the group consisting of: methoxy, ethoxy,
Figure PCTCN2018077229-appb-000014
Figure PCTCN2018077229-appb-000015
Figure PCTCN2018077229-appb-000015
在本发明的一个优选实施例中，所述F为
Figure PCTCN2018077229-appb-000016
In a preferred embodiment of the invention, the F is
Figure PCTCN2018077229-appb-000016
在本发明的一个具体实施方式中，所述制备方法的反应为：In a specific embodiment of the invention, the reaction of the preparation method is:
Figure PCTCN2018077229-appb-000017
Figure PCTCN2018077229-appb-000017
本发明另一方面还提供一种上述方法制备的Y型分支的亲水性聚合物羧酸衍生物，其具有如下结构：Another aspect of the present invention provides a Y-branched hydrophilic polymer carboxylic acid derivative prepared by the above method, which has the following structure:
Figure PCTCN2018077229-appb-000018
Figure PCTCN2018077229-appb-000018
其中，P a、P b、X 1、X 2、X 3具有本发明上述定义。 Among them, P a , P b , X 1 , X 2 and X 3 have the above definitions of the present invention.
所述羧酸衍生物的分子量为15-50KDa(具体可为15、16、18、20、22、24、26、28、30、32、34、36、38、40、42、44、46、48或50KDa)。The carboxylic acid derivative has a molecular weight of 15-50 KDa (specifically 15, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48 or 50KDa).
在本发明的一个实施例中，所述的P a和P b独立地选自：聚乙二醇、聚丙二醇、聚乙烯醇、聚四氢呋喃、聚环氧丙烷、聚环氧丁烷、聚氧杂环丁烷和聚丙烯吗啉中的一种或多种的共聚物的残基。 In one embodiment of the invention, the P a and P b are independently selected from the group consisting of polyethylene glycol, polypropylene glycol, polyvinyl alcohol, polytetrahydrofuran, polypropylene oxide, polybutylene oxide, polyoxygen A residue of a copolymer of one or more of a heterocyclic butane and a polypropylene morpholine.
在本发明的一个优选实施例中，所述的P a和/或P b为聚乙二醇残基。 In a preferred embodiment of the invention, the P a and/or P b are polyethylene glycol residues.
在本发明的一个实施例中，所述的P a为聚乙二醇残基，其结构为R a-O-(CH 2CH 2O) m-，R a选自：H、C 1-6烷基、C 3-6环烷基和C 6-10环烷基，m为170-565的整数。 In one embodiment of the present invention, the P a is a polyethylene glycol residue having a structure of R a —O—(CH 2 CH 2 O) m —, and R a is selected from the group consisting of: H, C 1- 6 alkyl, C 3-6 cycloalkyl and C 6-10 cycloalkyl, m is an integer from 170 to 565.
在本发明的一个实施例中，所述R a选自：H、甲基、乙基、异丙基、环丙基、环丁基、环己基和苄基。 In one embodiment of the present invention, said R a is selected from: H, methyl, ethyl, isopropyl, cyclopropyl, cyclobutyl, cyclohexyl, and benzyl.
在本发明的一个优选实施例中，所述R a为H或甲基。 In a preferred embodiment of the invention, said R a is H or methyl.
在本发明的一个实施例中，所述的P a为甲氧基聚乙二醇残基，其结构为CH 3O-(CH 2CH 2O) m-，m为170-565的整数。 In one embodiment of the invention, the P a is a methoxypolyethylene glycol residue having the structure CH 3 O-(CH 2 CH 2 O) m -, and m is an integer from 170 to 565.
在本发明的一个实施例中，所述的P a的分子量为7.5-25KDa(具体可为7.5、8、9、10、11、12、13、14、15、16、17、18、19、20、21、22、23、24或25KDa)。 In one embodiment of the present invention, the P a has a molecular weight of 7.5 to 25 KDa (specifically, 7.5, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24 or 25KDa).
在本发明的一个实施例中，所述的P b为聚乙二醇残基，其结构为R b-O-(CH 2CH 2O) n-，R b选自：H、C 1-6烷基、C 3-6环烷基和C 6-10环烷基，n为170-565的整数。 In one embodiment of the present invention, the P b is a polyethylene glycol residue having a structure of R b —O—(CH 2 CH 2 O) n —, and R b is selected from the group consisting of: H, C 1- 6 alkyl, C 3-6 cycloalkyl and C 6-10 cycloalkyl, n being an integer from 170 to 565.
在本发明的一个实施例中，所述R b选自：H、甲基、乙基、异丙基、环丙基、环丁基、环己基和苄基。 In one embodiment of the invention, the R b is selected from the group consisting of H, methyl, ethyl, isopropyl, cyclopropyl, cyclobutyl, cyclohexyl and benzyl.
在本发明的一个优选实施例中，所述R b为H或甲基。 In a preferred embodiment of the invention, said R b is H or methyl.
在本发明的一个实施例中，所述的P b为甲氧基聚乙二醇残基，其结构为CH 3O-(CH 2CH 2O) n-，n为170-565 的整数。 In one embodiment of the invention, the P b is a methoxypolyethylene glycol residue having the structure CH 3 O-(CH 2 CH 2 O) n -, n being an integer from 170 to 565.
在本发明的一个实施例中，所述的P b的分子量可为7.5-25KDa(具体可为7.5、8、9、10、11、12、13、14、15、16、17、18、19、20、21、22、23、24或25KDa)。 In one embodiment of the present invention, the molecular weight of P b may be 7.5-25 KDa (specifically, 7.5, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 , 20, 21, 22, 23, 24 or 25KDa).
在本发明的一个实施例中，所述Y型分支的亲水性聚合物羧酸衍生物为Y型分支的聚乙二醇羧酸衍生物，其具有如下结构：In one embodiment of the present invention, the Y-branched hydrophilic polymer carboxylic acid derivative is a Y-branched polyethylene glycol carboxylic acid derivative having the following structure:
Figure PCTCN2018077229-appb-000019
Figure PCTCN2018077229-appb-000019
在本发明的一个实施例中，所述X 1选自：单键、-CH 2-、-CH 2CH 2-、-CH 2CH 2CH 2-、-CH 2CH 2CH 2CH 2-、-CH 2CH 2CH 2CH 2CH 2-、-CH(CH 3)-、-CH 2CH(CH 3)-、-CH 2CH 2CH(CH 3)-、-CH 2CH 2CH 2CH(CH 3)-、-CH 2CH 2CH 2CH 2CH(CH 3)-、-CH 2CH 2CH 2CH 2CH 2CH(CH 3)-、-(CH 2) iO-和-(CH 2) iCO-中的一种或多种的组合，i为0-5的整数(如0、1、2、3、4或5)。 In one embodiment of the invention, the X 1 is selected from the group consisting of: a single bond, -CH 2 -, -CH 2 CH 2 -, -CH 2 CH 2 CH 2 -, -CH 2 CH 2 CH 2 CH 2 - , -CH 2 CH 2 CH 2 CH 2 CH 2 -, -CH(CH 3 )-, -CH 2 CH(CH 3 )-, -CH 2 CH 2 CH(CH 3 )-, -CH 2 CH 2 CH 2 CH(CH 3 )-, -CH 2 CH 2 CH 2 CH 2 CH(CH 3 )-, -CH 2 CH 2 CH 2 CH 2 CH 2 CH(CH 3 )-, -(CH 2 ) i O- and - i CO- combination of one or more of (2 CH), i is an integer (e.g., 3, 4 or 5) 0-5.
在本发明的一个优选实施例中，所述X 1为-CH 2CH 2-。 In a preferred embodiment of the invention, said X 1 is -CH 2 CH 2 -.
在本发明的一个实施例中，所述X 3选自：单键、-CH 2-、-CH 2CH 2-、-CH 2CH 2CH 2-、-CH 2CH 2CH 2CH 2-、-CH 2CH 2CH 2CH 2CH 2-、-CH(CH 3)-、-CH 2CH(CH 3)-、-CH 2CH 2CH(CH 3)-、-CH 2CH 2CH 2CH(CH 3)-、-CH 2CH 2CH 2CH 2CH(CH 3)-、-CH 2CH 2CH 2CH 2CH 2CH(CH 3)-、-(CH 2) iO-和-(CH 2) iCO-中的一种或多种的组合，i为0-5的整数(如0、1、2、3、4或5)。 In one embodiment of the invention, the X 3 is selected from the group consisting of: a single bond, -CH 2 -, -CH 2 CH 2 -, -CH 2 CH 2 CH 2 -, -CH 2 CH 2 CH 2 CH 2 - , -CH 2 CH 2 CH 2 CH 2 CH 2 -, -CH(CH 3 )-, -CH 2 CH(CH 3 )-, -CH 2 CH 2 CH(CH 3 )-, -CH 2 CH 2 CH 2 CH(CH 3 )-, -CH 2 CH 2 CH 2 CH 2 CH(CH 3 )-, -CH 2 CH 2 CH 2 CH 2 CH 2 CH(CH 3 )-, -(CH 2 ) i O- And a combination of one or more of -(CH 2 ) i CO-, i is an integer from 0 to 5 (eg, 0, 1, 2, 3, 4 or 5).
在本发明的一个优选实施例中，所述X 3为-CH 2-。 In a preferred embodiment of the invention, said X 3 is -CH 2 -.
在本发明的一个实施例中，所述X 2为
Figure PCTCN2018077229-appb-000020
In an embodiment of the invention, the X 2 is
Figure PCTCN2018077229-appb-000020
在本发明的一个实施例中，所述羧酸衍生物具有如下结构：In one embodiment of the invention, the carboxylic acid derivative has the structure:
Figure PCTCN2018077229-appb-000021
Figure PCTCN2018077229-appb-000021
在本发明的一个实施例中，所述R 3选自：-H、-CH 3、-CH 2CH(CH 3) 2、-CH(CH 3)CH 2CH 3、-CH(CH 3) 2、
Figure PCTCN2018077229-appb-000022
-CH 2CH 2SCH 3-、-CH 2OH、-CH 2SH、-CH(OH)CH 3、
Figure PCTCN2018077229-appb-000023
-CH 2COOH和-CH 2CH 2COOH。 In one embodiment of the invention, the R 3 is selected from the group consisting of: -H, -CH 3 , -CH 2 CH(CH 3 ) 2 , -CH(CH 3 )CH 2 CH 3 , -CH(CH 3 ) 2 ,
Figure PCTCN2018077229-appb-000022
-CH 2 CH 2 SCH 3 -, -CH 2 OH, -CH 2 SH, -CH(OH)CH 3 ,
Figure PCTCN2018077229-appb-000023
-CH 2 COOH and -CH 2 CH 2 COOH.
在本发明的一个优选实施例中，所述R 3为-H或-CH 3。 In a preferred embodiment of the invention, said R 3 is -H or -CH 3 .
在本发明的一个实施例中，所述m与n为相等的整数。In one embodiment of the invention, the m and n are equal integers.
本发明另一方面还提供一种基于上述羧酸衍生的Y型分支的亲水性聚合物衍生物，其具有如下结构：Another aspect of the present invention also provides a Y-branched hydrophilic polymer derivative derived from the above carboxylic acid, which has the following structure:
Figure PCTCN2018077229-appb-000024
Figure PCTCN2018077229-appb-000024
其中，P a、P b、X 1、X 2、X 3具有本发明上述定义， Wherein P a , P b , X 1 , X 2 , X 3 have the above definitions of the invention,
X 4为连接基团，选自：-(CH 2) j-、-(CH 2) jO-、-(CH 2) jS-、-(CH 2) jCO-、-(CH 2) jNH-、-(CH 2) jCONH-和-(CH 2) jNHCO-中一种或多种的组合，j为0-10的整数， X 4 is a linking group selected from the group consisting of: -(CH 2 ) j -, -(CH 2 ) j O-, -(CH 2 ) j S-, -(CH 2 ) j CO-, -(CH 2 ) a combination of one or more of j NH-, -(CH 2 ) j CONH- and -(CH 2 ) j NHCO-, j is an integer from 0 to 10,
Q为端基，选自：C 1-6的烷氧基、羟基、氨基、羧基、巯基、酯基、酮基、醛基、邻二硫吡啶基、叠氮基、酰肼基、炔基、硅烷基、马来酰亚胺基和琥珀酰亚胺基。 Q is a terminal group selected from: C 1-6 alkoxy, hydroxy, amino, carboxy, thiol, ester, keto, aldehyde, o-dithiopyridyl, azide, hydrazide, alkynyl , silyl, maleimide and succinimide groups.
在本发明的一个实施例中，所述衍生物的分子量可为15-50KDa(具体可为15、16、18、20、22、24、26、28、30、32、34、36、38、40、42、44、46、48或50KDa)。In one embodiment of the invention, the derivative may have a molecular weight of 15-50 KDa (specifically 15, 16, 16, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48 or 50 KDa).
在本发明的一个实施例中，所述的P a和P b独立地选自：聚乙二醇、聚丙二醇、聚乙烯醇、聚四氢呋喃、聚环氧丙烷、聚环氧丁烷、聚氧杂环丁烷和聚丙烯吗啉中的一种或多种的共聚物的残基。 In one embodiment of the invention, the P a and P b are independently selected from the group consisting of polyethylene glycol, polypropylene glycol, polyvinyl alcohol, polytetrahydrofuran, polypropylene oxide, polybutylene oxide, polyoxygen A residue of a copolymer of one or more of a heterocyclic butane and a polypropylene morpholine.
在本发明的一个优选实施例中，所述的P a和/或P b为聚乙二醇残基。 In a preferred embodiment of the invention, the P a and/or P b are polyethylene glycol residues.
在本发明的一个实施例中，所述的P a为聚乙二醇残基，其结构为R a-O-(CH 2CH 2O) m-，R a选自：H、C 1-6烷基、C 3-6环烷基和C 6-10环烷基，m为170-565的整数。 In one embodiment of the present invention, the P a is a polyethylene glycol residue having a structure of R a —O—(CH 2 CH 2 O) m —, and R a is selected from the group consisting of: H, C 1- 6 alkyl, C 3-6 cycloalkyl and C 6-10 cycloalkyl, m is an integer from 170 to 565.
在本发明的一个实施例中，所述R a选自：H、甲基、乙基、异丙基、环丙基、环丁基、环己基和苄基。 In one embodiment of the present invention, said R a is selected from: H, methyl, ethyl, isopropyl, cyclopropyl, cyclobutyl, cyclohexyl, and benzyl.
在本发明的一个优选实施例中，所述R a为H或甲基。 In a preferred embodiment of the invention, said R a is H or methyl.
在本发明的一个实施例中，所述的P a为甲氧基聚乙二醇残基，其结构为CH 3O-(CH 2CH 2O) m-，m为170-565的整数。 In one embodiment of the invention, the P a is a methoxypolyethylene glycol residue having the structure CH 3 O-(CH 2 CH 2 O) m -, and m is an integer from 170 to 565.
在本发明的一个实施例中，所述的P a的分子量为7.5-25KDa(具体可为7.5、8、9、10、11、12、13、14、15、16、17、18、19、20、21、22、23、24或25KDa)。 In one embodiment of the present invention, the P a has a molecular weight of 7.5 to 25 KDa (specifically, 7.5, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24 or 25KDa).
在本发明的一个实施例中，所述的P b为聚乙二醇残基，其结构为R b-O-(CH 2CH 2O) n-，R b选自：H、C 1-6烷基、C 3-6环烷基和C 6-10环烷基，n为170-565的整数。 In one embodiment of the present invention, the P b is a polyethylene glycol residue having a structure of R b —O—(CH 2 CH 2 O) n —, and R b is selected from the group consisting of: H, C 1- 6 alkyl, C 3-6 cycloalkyl and C 6-10 cycloalkyl, n being an integer from 170 to 565.
在本发明的一个实施例中，所述R b选自：H、甲基、乙基、异丙基、环丙基、环丁基、环己基和苄基。 In one embodiment of the invention, the R b is selected from the group consisting of H, methyl, ethyl, isopropyl, cyclopropyl, cyclobutyl, cyclohexyl and benzyl.
在本发明的一个优选实施例中，所述R b为H或甲基。 In a preferred embodiment of the invention, said R b is H or methyl.
在本发明的一个实施例中，所述的P b为甲氧基聚乙二醇残基，其结构为CH 3O-(CH 2CH 2O) n-，n为170-565的整数。 In one embodiment of the present invention, the P b using methoxy polyethylene glycol residue having the structure CH 3 O- (CH 2 CH 2 O) n -, n is an integer of 170-565.
在本发明的一个实施例中，所述的P b的分子量可为7.5-25KDa(具体可为7.5、8、9、10、11、12、13、14、15、16、17、18、19、20、21、22、23、24或25KDa)。 In one embodiment of the present invention, the molecular weight of P b may be 7.5-25 KDa (specifically, 7.5, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 , 20, 21, 22, 23, 24 or 25KDa).
在本发明的一个实施例中，所述Y型分支的亲水性聚合物衍生物为Y型分支的聚乙二醇衍生物，其具有如下结构：In one embodiment of the invention, the Y-branched hydrophilic polymer derivative is a Y-branched polyethylene glycol derivative having the following structure:
Figure PCTCN2018077229-appb-000025
Figure PCTCN2018077229-appb-000025
在本发明的一个实施例中，所述X 4选自：单键、-CH 2-、-CH 2CH 2-、-CH 2CH 2CH 2-、-CH 2CH 2CH 2CH 2-、-CH 2CH 2CH 2CH 2CH 2-、-(CH 2) jCO-、-(CH 2) jNH-、-(CH 2) jCONH-和-(CH 2) jNHCO-中一种或多种的组合，j为0-5的整数(如0、1、2、3、4或5)。 In one embodiment of the present invention, the X 4 is selected from: a single bond, -CH 2 -, - CH 2 CH 2 -, - CH 2 CH 2 CH 2 -, - CH 2 CH 2 CH 2 CH 2 - , -CH 2 CH 2 CH 2 CH 2 CH 2 -, -(CH 2 ) j CO-, -(CH 2 ) j NH-, -(CH 2 ) j CONH- and -(CH 2 ) j NHCO- A combination of one or more, j is an integer from 0 to 5 (eg, 0, 1, 2, 3, 4, or 5).
在本发明的一个优选实施例中，所述X 4选自：单键、-CH 2-、-CH 2CH 2-、-CH 2CH 2CH 2-、-CH 2NH-、-CH 2CH 2NH-、-CH 2CH 2CH 2NH-、-CH 2CONH-、-CH 2CH 2CONH-和-CH 2CH 2CH 2CONH-中一种或多种的组合。 In a preferred embodiment of the invention, the X 4 is selected from the group consisting of: a single bond, -CH 2 -, -CH 2 CH 2 -, -CH 2 CH 2 CH 2 -, -CH 2 NH-, -CH 2 CH 2 NH -, - CH 2 CH 2 CH 2 NH -, - CH 2 CONH -, - CH 2 CH 2 CONH- , and combinations -CH 2 CH 2 CH 2 CONH- of one or more.
在本发明的一个实施例中，所述Q选自：-OH、-SH、
Figure PCTCN2018077229-appb-000026
-NH 2、-COOH、-CHO、
Figure PCTCN2018077229-appb-000027
Figure PCTCN2018077229-appb-000028
和-N 3。 In an embodiment of the invention, the Q is selected from the group consisting of: -OH, -SH,
Figure PCTCN2018077229-appb-000026
-NH 2 , -COOH, -CHO,
Figure PCTCN2018077229-appb-000027
Figure PCTCN2018077229-appb-000028
And -N 3 .
在本发明的一个优选实施例中，所述Y型分支的亲水性聚合物衍生物具有如下结构：In a preferred embodiment of the invention, the Y-branched hydrophilic polymer derivative has the following structure:
Figure PCTCN2018077229-appb-000029
Figure PCTCN2018077229-appb-000029
在本发明的一个实施例中，所述R 3选自：-H、-CH 3、-CH 2CH(CH 3) 2、-CH(CH 3)CH 2CH 3、-CH(CH 3) 2、
Figure PCTCN2018077229-appb-000030
-CH 2CH 2SCH 3-、-CH 2OH、-CH 2SH、-CH(OH)CH 3、
Figure PCTCN2018077229-appb-000031
-CH 2COOH和-CH 2CH 2COOH。 In one embodiment of the invention, the R 3 is selected from the group consisting of: -H, -CH 3 , -CH 2 CH(CH 3 ) 2 , -CH(CH 3 )CH 2 CH 3 , -CH(CH 3 ) 2 ,
Figure PCTCN2018077229-appb-000030
-CH 2 CH 2 SCH 3 -, -CH 2 OH, -CH 2 SH, -CH(OH)CH 3 ,
Figure PCTCN2018077229-appb-000031
-CH 2 COOH and -CH 2 CH 2 COOH.
在本发明的一个优选实施例中，所述R 3为-H或-CH 3。 In a preferred embodiment of the invention, said R 3 is -H or -CH 3 .
在本发明的一个实施例中，所述m与n为相等的整数。In one embodiment of the invention, the m and n are equal integers.
本发明另一方面还提供一种上述Y型分支的亲水聚合物衍生物的制备方法，其包括上述Y型分支的亲水聚合物羧酸衍生物的制备方法步骤。Another aspect of the present invention provides a process for producing the above Y-branched hydrophilic polymer derivative, which comprises the steps of the preparation method of the above Y-branched hydrophilic polymer carboxylic acid derivative.
本发明另一方面还提供一种上述Y型分支的亲水性聚合物羧酸衍生物、Y型分支的亲水聚合物衍生物在修饰药物中的应用。Another aspect of the present invention provides a use of the above Y-branched hydrophilic polymer carboxylic acid derivative, Y-branched hydrophilic polymer derivative in a modified drug.
本发明另一方面还提供一种本发明上述Y型分支的亲水性聚合物羧酸衍生物、Y型分支的亲水聚合物衍生物与药物的结合物。Another aspect of the present invention provides a combination of the above Y-branched hydrophilic polymer carboxylic acid derivative, Y-branched hydrophilic polymer derivative and a drug of the present invention.
在本发明的一个实施例中，所述药物选自：氨基酸、多肽、蛋白质、糖类、有机酸、生物碱、黄酮类、醌类、萜类、苯丙素酚类、甾体和甙类药物。In one embodiment of the invention, the drug is selected from the group consisting of amino acids, polypeptides, proteins, sugars, organic acids, alkaloids, flavonoids, terpenoids, terpenoids, phenylpropanoid phenols, steroids, and steroids. drug.
本发明另一方面还提供一种本发明上述Y型分支的亲水性聚合物羧酸衍生物的制备方法在制备上述Y型分支的亲水性聚合物羧酸衍生物的药物组合物中的应用。Another aspect of the present invention provides a method for producing the above-mentioned Y-branched hydrophilic polymer carboxylic acid derivative of the present invention, in the pharmaceutical composition for preparing the above-mentioned Y-branched hydrophilic polymer carboxylic acid derivative application.
本发明另一方面还提供一种本发明上述Y型分支的亲水性聚合物羧酸衍生物的制备方法在制备上述Y型分支的亲水聚合物衍生物、及其药物结合物中的应用。Another aspect of the present invention provides a method for preparing the above-mentioned Y-branched hydrophilic polymer carboxylic acid derivative of the present invention, in the preparation of the above Y-branched hydrophilic polymer derivative, and a pharmaceutical combination thereof .
本发明另一方面还提供一种包括本发明上述结合物及任选的药学上可接受的载体或赋形剂的药物组合物。Another aspect of the invention also provides a pharmaceutical composition comprising the above combination of the invention and, optionally, a pharmaceutically acceptable carrier or excipient.
本发明另一方面还提供一种本发明上述Y型分支的亲水性聚合物羧酸衍生物、Y型分支的聚乙二醇衍生物、及其药物结合物和药物组合物在制备治疗疾病的药物中的应用。Another aspect of the present invention provides a Y-branched hydrophilic polymer carboxylic acid derivative, a Y-branched polyethylene glycol derivative, and a pharmaceutical conjugate thereof and a pharmaceutical composition thereof, in the preparation of a therapeutic disease The application of the drug.
本发明提供的Y型分支的亲水性聚合物羧酸衍生物(特别是高纯度的高分子量的Y型分支聚乙二醇羧酸衍生物)的制备方法，制备步骤简单，但反应后产物分离容易，分离成本低，产品纯度和收率高，利于后续基于该羧酸衍生物制备的其他衍生物及其药物结合物的制备，利于工业放大和商业应用。制备得到的Y型分支的亲水性聚合物羧酸衍生物(特别是高分子量的Y型分支聚乙二醇羧酸衍生物)产品纯度高，商业应用价值高，特别是在制备预防和/或治疗疾病的药物中的应用。The preparation method of the Y-branched hydrophilic polymer carboxylic acid derivative (especially the high-purity high molecular weight Y-branched polyethylene glycol carboxylic acid derivative) provided by the invention has simple preparation steps but the product after the reaction The separation is easy, the separation cost is low, the product purity and the yield are high, and the preparation of other derivatives based on the preparation of the carboxylic acid derivative and the drug combination thereof is facilitated, which is advantageous for industrial amplification and commercial application. The prepared Y-branched hydrophilic polymer carboxylic acid derivative (especially the high molecular weight Y-branched polyethylene glycol carboxylic acid derivative) has high purity and high commercial value, especially in the preparation of prevention and/or Or the application of drugs for the treatment of diseases.
附图说明DRAWINGS
图1所示为本发明实施例1提供的柱分前粗品的GFC色谱图。Figure 1 is a GFC chromatogram of the crude product before column separation provided in Example 1 of the present invention.
图2所示为本发明实施例1提供的收集起点GFC色谱图。2 is a chromatogram of a collection starting point GFC provided in Example 1 of the present invention.
图3所示为本发明实施例1提供的峰值点GFC色谱图。Fig. 3 is a view showing a peak point GFC chromatogram provided in Example 1 of the present invention.
图4所示为本发明实施例1提供的柱分后产品的GFC色谱图。4 is a GFC chromatogram of the post-column product provided in Example 1 of the present invention.
图5所示为本发明实施例2提供的柱分前粗品的GFC色谱图。Fig. 5 is a GFC chromatogram of the crude product before column separation according to Example 2 of the present invention.
图6所示为本发明实施例2提供的收集起点GFC色谱图。Figure 6 is a chromatogram of the collection starting point GFC provided in Example 2 of the present invention.
图7所示为本发明实施例2提供的峰值点GFC色谱图。Fig. 7 is a view showing a peak point GFC chromatogram provided in Example 2 of the present invention.
图8所示为本发明实施例2提供的柱分后产品的GFC色谱图。Figure 8 is a GFC chromatogram of the post-column product provided in Example 2 of the present invention.
图9所示为本发明实施例3提供的柱分前粗品的GFC色谱图。Figure 9 is a GFC chromatogram of the pre-column crude product provided in Example 3 of the present invention.
图10所示为本发明实施例3提供的收集起点GFC色谱图。Figure 10 is a chromatogram of the collection starting point GFC provided in Example 3 of the present invention.
图11所示为本发明实施例3提供的峰值点GFC色谱图。Figure 11 is a view showing a peak point GFC chromatogram provided in Example 3 of the present invention.
图12所示为本发明实施例3提供的柱分后产品的GFC色谱图。Figure 12 is a GFC chromatogram of the post-column product provided in Example 3 of the present invention.
图13所示为本发明实施例4提供的柱分前粗品的GFC色谱图。Figure 13 is a GFC chromatogram of the crude product before column separation according to Example 4 of the present invention.
图14所示为本发明实施例4提供的收集起点GFC色谱图。Figure 14 is a chromatogram of the collection starting point GFC provided in Example 4 of the present invention.
图15所示为本发明实施例4提供的峰值点GFC色谱图。Figure 15 is a view showing a peak point GFC chromatogram provided in Example 4 of the present invention.
图16所示为本发明实施例4提供的柱分后产品的GFC色谱图。Figure 16 is a GFC chromatogram of the post-column product provided in Example 4 of the present invention.
图17所示为本发明实施例5提供的柱分前粗品的GFC色谱图。Figure 17 is a GFC chromatogram of the pre-column crude product provided in Example 5 of the present invention.
图18所示为本发明实施例5提供的收集起点GFC色谱图。Figure 18 is a chromatogram of the collection starting point GFC provided in Example 5 of the present invention.
图19所示为本发明实施例5提供的峰值点GFC色谱图。Fig. 19 is a view showing a peak point GFC chromatogram provided in Example 5 of the present invention.
图20所示为本发明实施例5提供的柱分后产品的GFC色谱图。Figure 20 is a GFC chromatogram of the post-column product provided in Example 5 of the present invention.
具体实施方式detailed description
除非另有定义，本发明中所使用的所有的技术和科学术语具有与本发明涉及领域的技术人员通常理解的相同的含义，如，Unless defined otherwise, all technical and scientific terms used in the present invention have the same meaning meaning
“烷基”指的是直链或支链的且不含不饱和键的烃链自由基，且其以单键与分子其他部分连接。C1-C6的烷基指含有1-6个碳原子的烷基，如甲基、乙基、正丙基、异丙基、正丁基、异丁基、叔丁基、正戊基、异戊基、新戊基、叔戊基、正己基、异己基等。如果烷基被环烷基取代，其相应为“环烷基烷基”自由基，如环丙基甲基、环丙基乙基、环丁基甲基、环戊基甲基、环己基甲基等。如果烷基被芳基取代，那么其相应为“芳烷基”自由基，如苄基、二苯甲基或苯乙基。如果烷基被杂环基取代，那么其相应为“杂环基烷基”自由基。"Alkyl" refers to a hydrocarbon chain radical that is linear or branched and free of unsaturated bonds, and which is attached to the rest of the molecule by a single bond. The C1-C6 alkyl group means an alkyl group having 1 to 6 carbon atoms, such as methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, t-butyl, n-pentyl, iso Amyl, neopentyl, tert-amyl, n-hexyl, isohexyl and the like. If the alkyl group is substituted by a cycloalkyl group, it is correspondingly a "cycloalkylalkyl" radical such as cyclopropylmethyl, cyclopropylethyl, cyclobutylmethyl, cyclopentylmethyl, cyclohexylmethyl, etc. . If the alkyl group is substituted by an aryl group, it is correspondingly an "aralkyl" radical such as benzyl, benzhydryl or phenethyl. If the alkyl group is substituted by a heterocyclic group, it is correspondingly a "heterocyclylalkyl" radical.
“烷氧基”指的是羟基中的氢被烷基取代后形成的取代基，C1-C6的烷氧基指含有1-6个碳原子的烷氧基，如甲氧基、乙氧基、丙氧基、丁氧基等。"Alkoxy" means a substituent formed by substituting a hydrogen in a hydroxy group with an alkyl group, and alkoxy group of C1-C6 means an alkoxy group having 1 to 6 carbon atoms, such as a methoxy group or an ethoxy group. , propoxy, butoxy, and the like.
“环烷基”指的是脂环烃，如含1至4个单环和/或稠环、含3-18个碳原子，优选3-10个碳原子，如环丙基、环己基或金刚烷基等，本发明中所述C3-C6的环烷基指含有3-6个碳原子的环烷基，如环丙基、环丁基、环戊基和环己基。"Cycloalkyl" means an alicyclic hydrocarbon such as containing from 1 to 4 monocyclic and/or fused rings containing from 3 to 18 carbon atoms, preferably from 3 to 10 carbon atoms, such as cyclopropyl, cyclohexyl or Adamantyl and the like, the C3-C6 cycloalkyl group in the present invention means a cycloalkyl group having 3 to 6 carbon atoms such as a cyclopropyl group, a cyclobutyl group, a cyclopentyl group and a cyclohexyl group.
本发明中所述的“取代的”基团是指在基团的一个或多个可用位点被一个或多个合适的基团取代，具体地如，取代的烷基，是指烷基中的一个或多个氢被一个或多个合适的基团取代，所述合适的基团如烷基(如C1-6的烷基，特别是C1-3的烷基，如甲基、乙基、丙基或异丙基)、烷氧基(如C1-6的烷氧基，特别是C1-3的烷氧基，如甲氧基、乙氧基或丙氧基)、烯基(如C1-6的烯基，特别是C1-3的烯基，如乙烯基)、炔基(如C1-6的炔基，特别是C1-3的炔基，如丙炔基)、环烷基(如C3-6的环烷基，如环丙基、环丁基、环戊基或环己基)、芳基(如C6-12的芳基，特别是苯基)、芳氧基(如苯氧基)、烷基芳基(如苄基)、杂环基(如C3-12的杂环基，其含1、2或3中杂原子，所述杂原子选自N、O和S原子中一种或多种)、卤素(F、Cl、Br或I)、氰基(-CN)、羟基(-OH)、硝基(-NO 2)、叠氮基(-N 3)、酰基(如烷酰基，特别是C1-6的烷酰基，如甲酰基、乙酰基等；或，酰胺基)、胺基(如伯胺基、仲胺基)、羧基(-COOH)、酯基等。 A "substituted" group as used in the present invention refers to a group substituted at one or more of the available sites by one or more suitable groups, specifically, for example, a substituted alkyl group, which refers to an alkyl group. One or more hydrogens are substituted by one or more suitable groups such as an alkyl group (e.g., a C1-6 alkyl group, particularly a C1-3 alkyl group such as a methyl group, an ethyl group). , propyl or isopropyl), alkoxy (such as C1-6 alkoxy, especially C1-3 alkoxy, such as methoxy, ethoxy or propoxy), alkenyl (such as An alkenyl group of C1-6, especially an alkenyl group of C1-3, such as a vinyl group, an alkynyl group (such as an alkynyl group of C1-6, especially an alkynyl group of C1-3, such as a propynyl group), a cycloalkyl group (such as a cycloalkyl group of C3-6, such as cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl), an aryl group (such as an aryl group of C6-12, especially a phenyl group), an aryloxy group (such as benzene) An oxy), an alkylaryl group (such as a benzyl group), a heterocyclic group (such as a heterocyclic group of C3-12, which contains a hetero atom in 1, 2 or 3, wherein the hetero atom is selected from the group consisting of N, O and S atoms One or more), halogen (F, Cl, Br or I), cyano (-CN), hydroxy (-OH), nitro (-NO 2 ), azide (-N 3 ), acyl (such as alkane Group, especially a C1-6 alkanoyl, such as formyl, acetyl and the like; or an amide group), amino (e.g., primary amino, secondary amino), a carboxyl group (-COOH), an ester group and the like.
另外，本发明中涉及的一些具体基团及其化学结构对应如下：羟基，-OH；氨基，-NH 2；羧基，
Figure PCTCN2018077229-appb-000032
巯基，-SH；酯基，
Figure PCTCN2018077229-appb-000033
(其中Q 1可为烷基、芳基或杂环基，如甲基、乙基、正丙基、叔丁基、马来酰亚胺基、琥珀酰亚胺基、
Figure PCTCN2018077229-appb-000034
Figure PCTCN2018077229-appb-000035
等)；酮基，
Figure PCTCN2018077229-appb-000036
(其中Q 2可为取代或未取代的烷基、芳基、杂环基，如取代或未取代的甲基、乙基、正丙基、
Figure PCTCN2018077229-appb-000037
等)；醛基，-CHO；邻二硫吡啶基，
Figure PCTCN2018077229-appb-000038
叠氮基，
Figure PCTCN2018077229-appb-000039
酰肼基，
Figure PCTCN2018077229-appb-000040
炔基，
Figure PCTCN2018077229-appb-000041
硅烷基，
Figure PCTCN2018077229-appb-000042
(其中Q 3可为相同或不同的烷基或烷氧基，如甲基、乙基、丙基、丁基、戊基、甲氧基、乙氧基、丙氧基、丁氧基等，优选地，Q 3均为甲基、乙基、正丙基、甲氧基、乙氧基、正丙氧基等)；马来酰亚胺基，
Figure PCTCN2018077229-appb-000043
琥珀酰亚胺基，
Figure PCTCN2018077229-appb-000044
In addition, some specific groups and chemical structures involved in the present invention correspond to the following: hydroxyl group, -OH; amino group, -NH 2 ; carboxyl group,
Figure PCTCN2018077229-appb-000032
Sulfhydryl, -SH; ester group,
Figure PCTCN2018077229-appb-000033
(wherein Q 1 may be an alkyl group, an aryl group or a heterocyclic group such as methyl, ethyl, n-propyl, t-butyl, maleimide, succinimidyl,
Figure PCTCN2018077229-appb-000034
Figure PCTCN2018077229-appb-000035
Ketone,
Figure PCTCN2018077229-appb-000036
(wherein Q 2 may be substituted or unsubstituted alkyl, aryl, heterocyclic, such as substituted or unsubstituted methyl, ethyl, n-propyl,
Figure PCTCN2018077229-appb-000037
Ethyl, -CHO; o-dithiopyridyl,
Figure PCTCN2018077229-appb-000038
Azido group,
Figure PCTCN2018077229-appb-000039
Acyl hydrazino,
Figure PCTCN2018077229-appb-000040
Alkynyl,
Figure PCTCN2018077229-appb-000041
Silyl group,
Figure PCTCN2018077229-appb-000042
(wherein Q 3 may be the same or different alkyl or alkoxy group, such as methyl, ethyl, propyl, butyl, pentyl, methoxy, ethoxy, propoxy, butoxy, etc. Preferably, Q 3 is methyl, ethyl, n-propyl, methoxy, ethoxy, n-propoxy, etc.; maleimide group,
Figure PCTCN2018077229-appb-000043
Succinimide group,
Figure PCTCN2018077229-appb-000044
本发明中关于连接基团的定义中，所述“组合”是指所列举的连接基团中的两个以上通过化学键键合连接后形成的基团，例如-(CH 2) j-与-(CH 2) jNHCO-的组合可为-(CH 2) jNHCO(CH 2) j-；具体地，如-CH 2-与-CH 2CH 2NHCO-的组合可为-CH 2CH 2NHCOCH 2-、-CH 2CH 2CH 2NHCO-。所述“组合”用于限定连接基团的化学结构，不涉及连接基团的制备步骤、组合顺序等。 In the present invention, the definition of the linking group, the "combination" refers to the group two linking groups listed in the above chemical bond formed by the engagement of the connection, for example - (CH 2) j - and - The combination of (CH 2 ) j NHCO- may be -(CH 2 ) j NHCO(CH 2 ) j -; specifically, the combination of -CH 2 - and -CH 2 CH 2 NHCO- may be -CH 2 CH 2 NHCOCH 2 -, -CH 2 CH 2 CH 2 NHCO-. The "combination" is used to define the chemical structure of the linking group, and does not involve the preparation steps, combination order, and the like of the linking group.
下面将结合本发明实施例，对本发明的技术方案进行清楚、完整地描述，显然，所描述的实施例仅是本发明一部分实施例，而不是全部的实施例。基于本发明中的实施例，本领域普通技术人员在没有付出创造性劳动前提下所获得的所有其他实施例，都属于本发明保护的范围。The technical solutions of the present invention will be described clearly and completely in conjunction with the embodiments of the present invention. It is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments obtained by a person of ordinary skill in the art based on the embodiments of the present invention without departing from the inventive scope are the scope of the present invention.
实施例1Y型分支的聚乙二醇-乙酸(分子量4000)的合成(现有技术)Synthesis of Y-branched polyethylene glycol-acetic acid (molecular weight 4000) of Example 1 (prior art)
Figure PCTCN2018077229-appb-000045
Figure PCTCN2018077229-appb-000045
参照专利CN1243779C实施例5的合成方法制备分子量为4000的Y型分支的聚乙二醇-乙酸：将10g分子量为2000的聚乙二醇单甲醚-氨基乙酸(mPEG-Gly)和10g分子量为2000的聚乙二醇单甲醚-羧乙酸丁二酰亚胺酯(mPEG-OCH 2CO-NHS)溶于200ml二氯甲烷，向溶液中加入2.5ml三乙胺，室温反应过夜，旋蒸浓缩溶剂，残余物加入乙醚，过滤收集沉淀，真空干燥，离子交换色谱柱纯化，凝胶过滤色谱(GFC)监测，以目标产物峰高超过5mv开始收集，低于5mv结束收集。 Preparation of a Y-branched polyethylene glycol-acetic acid having a molecular weight of 4000 according to the synthesis method of Example 5 of Patent CN1243779C: 10 g of polyethylene glycol monomethyl ether-aminoacetic acid (mPEG-Gly) having a molecular weight of 2000 and a molecular weight of 10 g 2000 polyethylene glycol monomethyl ether-carboxylic acid succinimide ester (mPEG-OCH 2 CO-NHS) dissolved in 200 ml of dichloromethane, 2.5 ml of triethylamine was added to the solution, and reacted at room temperature overnight, steamed The solvent was concentrated, the residue was added to diethyl ether, and the precipitate was collected by filtration, dried in vacuo, purified by ion-exchange chromatography column, and filtered by gel filtration chromatography (GFC), and collected at a peak height of the target product exceeding 5 mv, and collected at less than 5 mv.
柱分前粗品的GFC色谱图如图1所示，其中结果分析如表1所示：The GFC chromatogram of the crude product before column is shown in Figure 1. The results are shown in Table 1:
表1 分析结果表Table 1 Analysis Results Table
Figure PCTCN2018077229-appb-000046
Figure PCTCN2018077229-appb-000046
收集起点GFC色谱图如图2所示，其中结果分析如表2所示：The GHC chromatogram of the collection starting point is shown in Figure 2, and the results are analyzed as shown in Table 2:
表2 分析结果表Table 2 Analysis Results Table
Figure PCTCN2018077229-appb-000047
Figure PCTCN2018077229-appb-000047
峰值点GFC色谱图如图3所示，其中结果分析如表3所示：The peak point GFC chromatogram is shown in Figure 3, and the results are analyzed as shown in Table 3:
表3 分析结果表Table 3 Analysis Results Table
Figure PCTCN2018077229-appb-000048
Figure PCTCN2018077229-appb-000048
Figure PCTCN2018077229-appb-000049
Figure PCTCN2018077229-appb-000049
柱分后产品的GFC色谱图如图4所示，其中结果分析如表4所示：The GFC chromatogram of the product after column separation is shown in Figure 4, and the results are shown in Table 4:
表4 分析结果表Table 4 Analysis Results Table
Figure PCTCN2018077229-appb-000050
Figure PCTCN2018077229-appb-000050
由上述图谱和分析结果可以看出，在低分子量情况下，应用现有技术即可得到纯度较高的粗品，相应地，纯化过程也很顺利，收集起点处的目标产物纯度已达100％，因此没有必要进行工艺改进。It can be seen from the above-mentioned map and analysis results that in the case of low molecular weight, the crude product with higher purity can be obtained by using the prior art, and accordingly, the purification process is also smooth, and the purity of the target product at the collection starting point has reached 100%. Therefore, no process improvement is necessary.
实施例2Y型分支的聚乙二醇-乙酸(分子量20000)的合成(现有技术)Example 2 Synthesis of Y-branched polyethylene glycol-acetic acid (molecular weight 20000) (prior art)
参照专利CN1243779C实施例5的合成方法制备分子量为20000的Y型分支的聚乙二醇-乙酸：将10g分子量为10000的聚乙二醇单甲醚-氨基乙酸(mPEG-Gly)和10g分子量为10000的聚乙二醇单甲醚-羧乙酸丁二酰亚胺酯(mPEG-OCH 2CO-NHS)溶于200ml二氯甲烷，向溶液中加入0.11ml三乙胺，室温反应过夜，旋蒸浓缩溶剂，残余物加入乙醚，过滤收集沉淀，真空干燥，离子交换色谱柱纯化，GFC监测，以目标产物峰高超过5mv开始收集，低于5mv结束收集。 The Y-branched polyethylene glycol-acetic acid having a molecular weight of 20,000 was prepared by the synthesis method of Example 5 of Patent CN1243779C: 10 g of polyethylene glycol monomethyl ether-aminoacetic acid (mPEG-Gly) having a molecular weight of 10,000 and a molecular weight of 10 g 10000 of polyethylene glycol monomethyl ether-carboxyacetate succinimide ester (mPEG-OCH 2 CO-NHS) was dissolved in 200 ml of dichloromethane, 0.11 ml of triethylamine was added to the solution, and the reaction was carried out overnight at room temperature. The solvent was concentrated, the residue was added diethyl ether, and the precipitate was collected by filtration, dried in vacuo, purified by ion-exchange chromatography column, and monitored by GFC, and collected at a peak height of the target product exceeding 5 mv, and collected at less than 5 mv.
柱分前粗品GFC的色谱图如图5所示，其中结果分析如表5所示：The chromatogram of the crude GFC before column is shown in Figure 5, and the results are shown in Table 5:
表5 分析结果表Table 5 Analysis Results Table
Figure PCTCN2018077229-appb-000051
Figure PCTCN2018077229-appb-000051
收集起点的GFC色谱图如图6所示，其中结果分析如表6所示：The GFC chromatogram of the collection starting point is shown in Figure 6, and the results are analyzed as shown in Table 6:
表6 分析结果表Table 6 Analysis Results Table
Figure PCTCN2018077229-appb-000052
Figure PCTCN2018077229-appb-000052
峰值点GFC色谱图如图7所示，其中结果分析如表7所示：The peak point GFC chromatogram is shown in Figure 7, and the results are analyzed as shown in Table 7:
表7 分析结果表Table 7 Analysis Results Table
Figure PCTCN2018077229-appb-000053
Figure PCTCN2018077229-appb-000053
柱分后产品的GFC色谱图如图8所示，其中结果分析如表8所示：The GFC chromatogram of the post-column product is shown in Figure 8, and the results are shown in Table 8:
表8 分析结果表Table 8 Analysis Results Table
Figure PCTCN2018077229-appb-000054
Figure PCTCN2018077229-appb-000054
实施例3Y型分支的聚乙二醇-乙酸(分子量20000)的合成(本发明方法)Synthesis of the Y-branched polyethylene glycol-acetic acid (molecular weight 20000) of Example 3 (method of the present invention)
mPEG-Gly与mPEG-OCH 2CO-NHS反应的步骤参考实施例2，室温反应过夜后，加入BOC酸酐，反应3h，旋蒸浓缩溶剂，残余物加入乙醚，过滤收集沉淀，真空干燥，离子交换色谱柱纯化，GFC监测，以目标产物峰高超过5mv开始收集，低于5mv结束收集。 Step of reacting mPEG-Gly with mPEG-OCH 2 CO-NHS Referring to Example 2, after reacting at room temperature overnight, BOC anhydride was added, the reaction was carried out for 3 h, the solvent was concentrated by rotary evaporation, the residue was added diethyl ether, and the precipitate was collected by filtration, dried under vacuum, and ion exchange Column purification, GFC monitoring, starting with the peak height of the target product exceeding 5 mv, and collecting at less than 5 mv.
柱分前粗品的GFC色谱图如图9所示，其中结果分析如表9所示：The GFC chromatogram of the crude product before column is shown in Figure 9, and the results are shown in Table 9.
表9 分析结果表Table 9 Analysis Results Table
Figure PCTCN2018077229-appb-000055
Figure PCTCN2018077229-appb-000055
Figure PCTCN2018077229-appb-000056
Figure PCTCN2018077229-appb-000056
收集起点的GFC色谱图如图10所示，其中结果分析如表10所示：The GFC chromatogram of the collection starting point is shown in Figure 10, and the results are analyzed as shown in Table 10:
表10 分析结果表Table 10 Analysis Results Table
Figure PCTCN2018077229-appb-000057
Figure PCTCN2018077229-appb-000057
峰值点GFC色谱图如图11所示，其中结果分析如表11所示：The peak point GFC chromatogram is shown in Figure 11, and the results are analyzed as shown in Table 11:
表11 分析结果表Table 11 Analysis Results Table
Figure PCTCN2018077229-appb-000058
Figure PCTCN2018077229-appb-000058
柱分后产品的GFC色谱图如图12所示，其中结果分析如表12所示：The GFC chromatogram of the post-column product is shown in Figure 12, and the results are shown in Table 12:
表12 分析结果表Table 12 Analysis Results Table
Figure PCTCN2018077229-appb-000059
Figure PCTCN2018077229-appb-000059
由实施例2、3对比可以看出，在制备分子量20000的Y型分支的聚乙二醇-乙酸产品过程中，应用现有技术会对后期纯化过程产生一定不利的影响，在目标产物的浓度达到收集标准时纯度为89.2％，最终所得的产品纯度只能达到96.2％。不过，该分子量条件下，目标产物浓度达到峰值时仍能达到纯度100％，因此，若想通过现有技术得到纯度100％的产品，则需要在柱分过程中舍弃部分目标产物，继而影响收率。相比较，应用本发明方法后，则可以在柱分收集的起点处就得到纯度100％的目标产物，从而提高纯化效率。It can be seen from the comparison of Examples 2 and 3 that in the preparation of the Y-branched polyethylene glycol-acetic acid product having a molecular weight of 20,000, the application of the prior art may have a certain adverse effect on the later purification process, at the concentration of the target product. When the collection standard is reached, the purity is 89.2%, and the final product purity can only reach 96.2%. However, under the molecular weight condition, the purity of the target product reaches a peak value of 100%. Therefore, if a product with a purity of 100% is obtained by the prior art, it is necessary to discard some of the target products in the column separation process, and then affect the collection. rate. In comparison, after applying the method of the present invention, the target product having a purity of 100% can be obtained at the starting point of the column collection, thereby improving the purification efficiency.
实施例4Y型分支的聚乙二醇-乙酸(分子量44000)的合成(现有技术)Example 4 Synthesis of Y-branched polyethylene glycol-acetic acid (molecular weight 44000) (prior art)
参照专利CN1243779C实施例5的合成方法制备分子量为44000的Y型分支的聚乙二醇-乙酸：将10g分子量为22000的聚乙二醇单甲醚-氨基乙酸(mPEG-Gly)和10g分子量为22000的聚乙二醇单甲醚-羧乙酸丁二酰亚胺酯(mPEG-OCH 2CO-NHS)溶于200ml二氯甲烷，向溶液中加入0.23ml三乙胺，室温反应过夜，旋蒸浓缩溶剂，残余物加入乙醚，过滤收集沉淀，真空干燥，离子交换色谱柱纯化，GFC监测，以目标产物峰高超过5mv开始收集，低于5mv结束收集。 The Y-branched polyethylene glycol-acetic acid having a molecular weight of 44,000 was prepared by the synthesis method of Example 5 of Patent CN1243779C: 10g of polyethylene glycol monomethyl ether-aminoacetic acid (mPEG-Gly) having a molecular weight of 22000 and a molecular weight of 10g 22000 of polyethylene glycol monomethyl ether-carboxyacetate succinimide ester (mPEG-OCH 2 CO-NHS) was dissolved in 200 ml of dichloromethane, 0.23 ml of triethylamine was added to the solution, and the reaction was carried out overnight at room temperature. The solvent was concentrated, the residue was added diethyl ether, and the precipitate was collected by filtration, dried in vacuo, purified by ion-exchange chromatography column, and monitored by GFC, and collected at a peak height of the target product exceeding 5 mv, and collected at less than 5 mv.
柱分前粗品的GFC色谱图如图13所示，其中结果分析如表13所示：The GFC chromatogram of the crude product before column separation is shown in Figure 13, and the results are shown in Table 13:
表13 分析结果表Table 13 Analysis Results Table
Figure PCTCN2018077229-appb-000060
Figure PCTCN2018077229-appb-000060
收集起点的GFC色谱图如图14所示，其中结果分析如表14所示：The GFC chromatogram of the collection starting point is shown in Figure 14, and the results are analyzed as shown in Table 14:
表14 分析结果表Table 14 Analysis Results Table
Figure PCTCN2018077229-appb-000061
Figure PCTCN2018077229-appb-000061
峰值点GFC色谱图如图15所示，其中结果分析如表15所示：The peak point GFC chromatogram is shown in Figure 15, and the results are analyzed as shown in Table 15:
表15 分析结果表Table 15 Analysis Results Table
Figure PCTCN2018077229-appb-000062
Figure PCTCN2018077229-appb-000062
Figure PCTCN2018077229-appb-000063
Figure PCTCN2018077229-appb-000063
柱分后产品的GFC色谱图如图16所示，其中结果分析如表16所示：The GFC chromatogram of the post-column product is shown in Figure 16, and the results are analyzed as shown in Table 16:
表16 分析结果表Table 16 Analysis Results Table
Figure PCTCN2018077229-appb-000064
Figure PCTCN2018077229-appb-000064
实施例5Y型分支的聚乙二醇-乙酸(分子量44000)的合成(本发明方法)Synthesis of the Y-branched polyethylene glycol-acetic acid (molecular weight 44000) of Example 5 (method of the present invention)
mPEG-Gly与mPEG-OCH 2CO-NHS反应的步骤参考实施例4，室温反应过夜后，加入BOC酸酐，反应3h，旋蒸浓缩溶剂，残余物加入乙醚，过滤收集沉淀，真空干燥，离子交换色谱柱纯化，GFC监测，以目标产物峰高超过5mv开始收集，低于5mv结束收集。 Step of reacting mPEG-Gly with mPEG-OCH 2 CO-NHS Referring to Example 4, after reacting at room temperature overnight, adding BOC anhydride, reacting for 3 hours, concentrating the solvent by rotary evaporation, adding the residue to diethyl ether, collecting the precipitate by filtration, vacuum drying, ion exchange Column purification, GFC monitoring, starting with the peak height of the target product exceeding 5 mv, and collecting at less than 5 mv.
柱分前粗品的GFC色谱图如图17所示，其中结果分析如表17所示：The GFC chromatogram of the crude product before column separation is shown in Figure 17, and the results are shown in Table 17:
表17 分析结果表Table 17 Analysis Results Table
Figure PCTCN2018077229-appb-000065
Figure PCTCN2018077229-appb-000065
收集起点的GFC色谱图如图18所示，其中结果分析如表18所示：The GFC chromatogram of the collection starting point is shown in Figure 18, and the results are analyzed as shown in Table 18:
表18 分析结果表Table 18 Analysis Results Table
Figure PCTCN2018077229-appb-000066
Figure PCTCN2018077229-appb-000066
峰值点GFC色谱图如图19所示，其中结果分析如表19所示：The peak point GFC chromatogram is shown in Figure 19, and the results are analyzed as shown in Table 19:
表19 分析结果表Table 19 Analysis Results Table
Figure PCTCN2018077229-appb-000067
Figure PCTCN2018077229-appb-000067
柱分后产品的GFC色谱图如图20所示，其中结果分析如表20所示：The GFC chromatogram of the post-column product is shown in Figure 20, and the results are analyzed as shown in Table 20:
表20 分析结果表Table 20 Analysis Results Table
Figure PCTCN2018077229-appb-000068
Figure PCTCN2018077229-appb-000068
由实施例4、5对比可以看出，在制备分子量44000的Y型分支的聚乙二醇-乙酸产品过程中，应用现有技术会对后期纯化过程产生严重影响，在目标产物的浓度达到收集标准时纯度为84.7％，峰值时纯度也只有97.2％，最终所得的产品纯度只能达到94.5％，使用该方法无法得到高纯产品。相比较，应用本发明方法后，则可以在柱分收集的起点处就得到纯度100％的目标产物，显著地提高了纯化的效果。It can be seen from the comparison of Examples 4 and 5 that in the preparation of the Y-branched polyethylene glycol-acetic acid product having a molecular weight of 44,000, the application of the prior art will have a serious impact on the later purification process, and the concentration of the target product is collected. The purity at the standard is 84.7%, and the purity at the peak is only 97.2%. The purity of the final product can only reach 94.5%. High purity products cannot be obtained by this method. In comparison, after applying the method of the present invention, a target product having a purity of 100% can be obtained at the starting point of column collection, which significantly improves the purification effect.
由以上实施例可以看出，现有技术对于产品纯化的影响会随着分子量的提高而逐渐增加，本发明的方法可以针对高分子量产品显著地提高纯化收率以及产品纯度。As can be seen from the above examples, the effect of the prior art on product purification will gradually increase with increasing molecular weight, and the method of the present invention can significantly improve purification yield and product purity for high molecular weight products.
以上所述仅为本发明的较佳实施例而已，并不用以限制本发明，凡在本发明的精神和原则之内，所作的任何修改、等同替换等，均应包含在本发明的保护范围之内。The above is only the preferred embodiment of the present invention, and is not intended to limit the present invention. Any modifications, equivalent substitutions, etc., which are within the spirit and principles of the present invention, should be included in the scope of the present invention. within.

Claims (23)

  1. 一种Y型分支的亲水性聚合物羧酸衍生物的制备方法，所述方法包括如下反应：A method for preparing a Y-branched hydrophilic polymer carboxylic acid derivative, the method comprising the following reaction:
    Figure PCTCN2018077229-appb-100001
    Figure PCTCN2018077229-appb-100001
    其中，P a和P b为相同或不同的亲水性聚合物残基， Wherein P a and P b are the same or different hydrophilic polymer residues,
    X 1和X 3为连接基团，独立地选自：-(CH 2) i-、
    Figure PCTCN2018077229-appb-100002
    、-(CH 2) iO-、-(CH 2) iS-和-(CH 2) iCO-中的一种或多种的组合，i为0-10的整数，     X 1 and X 3 are a linking group independently selected from: -(CH 2 ) i -,
    Figure PCTCN2018077229-appb-100002
    a combination of one or more of -(CH 2 ) i O-, -(CH 2 ) i S- and -(CH 2 ) i CO-, i is an integer from 0 to 10,
    X 2为连接基团，选自：-(CH 2) r-、-(CH 2) rO-、-(CH 2) rS-和
    Figure PCTCN2018077229-appb-100003
    中的一种或多种的组合，r为0-10的整数，     X 2 is a linking group selected from the group consisting of: -(CH 2 ) r -, -(CH 2 ) r O-, -(CH 2 ) r S- and
    Figure PCTCN2018077229-appb-100003
    a combination of one or more of the following, r is an integer from 0 to 10,
    F为端基，选自：C 1-6的取代或未取代的烷基、C 1-6的取代或未取代的烷氧基、
    Figure PCTCN2018077229-appb-100004
    Figure PCTCN2018077229-appb-100005
    F is a terminal group selected from: substituted or unsubstituted C 1-6 alkyl, C 1-6 substituted or unsubstituted alkoxy group,
    Figure PCTCN2018077229-appb-100004
    Figure PCTCN2018077229-appb-100005
    R 1和R 2独立地选自：-H、C 1-6取代或未取代的烷基、C 1-6取代或未取代的烷氧基、C 3-6取代或未取代的环烷基和C 4-10取代或未取代的亚烷基环烷基， R 1 and R 2 are independently selected from: -H, C 1-6 substituted or unsubstituted alkyl, C 1-6 substituted or unsubstituted alkoxy, C 3-6 substituted or unsubstituted cycloalkyl And a C 4-10 substituted or unsubstituted alkylene cycloalkyl group,
    R 3选自：-H、C 1-6取代或未取代的烷基、C 6-10取代或未取代的芳烷基和C 4-10取代或未取代的杂环烷基， R 3 is selected from the group consisting of: -H, a C 1-6 substituted or unsubstituted alkyl group, a C 6-10 substituted or unsubstituted aralkyl group, and a C 4-10 substituted or unsubstituted heterocycloalkyl group,
    反应结束后，加入酸酐，继续反应，分离纯化。After completion of the reaction, an acid anhydride is added, and the reaction is continued and separated and purified.
  2. 如权利要求1所述的制备方法，其特征在于，所述酸酐为有机酸酸酐，优选的，所述有机酸酸酐选自：二碳酸二叔丁酯、乙酸酐、丙酸酐、异丁酸酐、丁酸酐、苯甲酸酐和邻苯二甲酸酐中的一种或多种；和/或，The preparation method according to claim 1, wherein the acid anhydride is an organic acid anhydride, and preferably, the organic acid anhydride is selected from the group consisting of di-tert-butyl dicarbonate, acetic anhydride, propionic anhydride, isobutyric anhydride, One or more of butyric anhydride, benzoic anhydride, and phthalic anhydride; and/or,
    所述酸酐与反应物
    Figure PCTCN2018077229-appb-100006
    加入量的摩尔比为0.01-10:1；和/或，     The acid anhydride and reactant
    Figure PCTCN2018077229-appb-100006
    The molar ratio of the added amount is from 0.01 to 10:1; and/or,
    加入酸酐后反应时间为0.1-24小时；和/或，The reaction time after adding the acid anhydride is 0.1-24 hours; and/or,
    所述分离纯化步骤包括采用离子交换色谱分离纯化的步骤。The separation and purification step includes the step of separating and purifying using ion exchange chromatography.
  3. 如权利要求1或2所述的制备方法，其特征在于，所述Y型分支的亲水性聚合物羧酸衍生物的分子量为15-50KDa；和/或，The method according to claim 1 or 2, wherein the Y-branched hydrophilic polymer carboxylic acid derivative has a molecular weight of 15 to 50 KDa; and/or
    所述的P a选自：聚乙二醇、聚丙二醇、聚乙烯醇、聚四氢呋喃、聚环氧丙烷、聚环氧丁烷、聚氧杂环丁烷和聚丙烯吗啉中的一种或多种的共聚物的残基；和/或， The P a is selected from the group consisting of polyethylene glycol, polypropylene glycol, polyvinyl alcohol, polytetrahydrofuran, polypropylene oxide, polybutylene oxide, polyoxetane and polypropylene morpholine or a residue of a plurality of copolymers; and/or,
    所述的P a的分子量为7.5-25KDa；和/或， The molecular weight of P a is 7.5-25 KDa; and/or,
    所述的P b选自：聚乙二醇、聚丙二醇、聚乙烯醇、聚四氢呋喃、聚环氧丙烷、聚环氧丁烷、聚氧杂环丁烷和聚丙烯吗啉中的一种或多种的共聚物的残基；和/或， The P b is selected from the group consisting of polyethylene glycol, polypropylene glycol, polyvinyl alcohol, polytetrahydrofuran, polypropylene oxide, polybutylene oxide, polyoxetane and polypropylene morpholine or a residue of a plurality of copolymers; and/or,
    所述的P b的分子量为7.5-25KDa。 The P b has a molecular weight of 7.5 to 25 KDa.
  4. 如权利要求3所述的制备方法，其特征在于，所述的P a为聚乙二醇残基，其结构为R a-O-(CH 2CH 2O) m-，所述R a选自：H、甲基、乙基、异丙基、环丙基、环丁基、环己基和苄基，m为170-565的整数；和/或， The preparation method according to claim 3, wherein said P a is a polyethylene glycol residue having a structure of R a -O-(CH 2 CH 2 O) m -, said R a being selected From: H, methyl, ethyl, isopropyl, cyclopropyl, cyclobutyl, cyclohexyl and benzyl, m is an integer from 170 to 565; and/or,
    所述的P b为聚乙二醇残基，其结构为R b-O-(CH 2CH 2O) n-，R b选自：H、甲基、乙基、异丙基、环丙基、环丁基、环己基和苄基，n为170-565的整数。 The P b is a polyethylene glycol residue having a structure of R b —O—(CH 2 CH 2 O) n —, and R b is selected from the group consisting of: H, methyl, ethyl, isopropyl, cyclopropane Base, cyclobutyl, cyclohexyl and benzyl, n is an integer from 170 to 565.
  5. 如权利要求3所述的制备方法，其特征在于，所述R a为H或甲基；和/或，所述R b为H或甲基；和/或，所述m与n为相等的整数。 The method according to claim 3, wherein said R a is H or a methyl group; and/or said R b is H or a methyl group; and/or said m and n are equal. Integer.
  6. 如权利要求3所述的制备方法，其特征在于，所述Y型分支的亲水性聚合物羧酸衍生物为Y型分支的聚乙二醇羧酸衍生物，所述反应如下：The method according to claim 3, wherein the Y-branched hydrophilic polymer carboxylic acid derivative is a Y-branched polyethylene glycol carboxylic acid derivative, and the reaction is as follows:
    Figure PCTCN2018077229-appb-100007
    Figure PCTCN2018077229-appb-100007
  7. 如权利要求1或2所述的制备方法，其特征在于，所述X 1和X 3独立地选自：单键、-CH 2-、-CH 2CH 2-、-CH 2CH 2CH 2-、-CH 2CH 2CH 2CH 2-、-CH 2CH 2CH 2CH 2CH 2-、-CH(CH 3)-、-CH 2CH(CH 3)-、-CH 2CH 2CH(CH 3)-、-CH 2CH 2CH 2CH(CH 3)-、-CH 2CH 2CH 2CH 2CH(CH 3)-、-CH 2CH 2CH 2CH 2CH 2CH(CH 3)-、-(CH 2) iO-和-(CH 2) iCO-中的一种或多种的组合，i为0-5的整数；和/或， The process according to claim 1 or 2, wherein X 1 and X 3 are independently selected from the group consisting of: a single bond, -CH 2 -, -CH 2 CH 2 -, -CH 2 CH 2 CH 2 -, -CH 2 CH 2 CH 2 CH 2 -, -CH 2 CH 2 CH 2 CH 2 CH 2 -, -CH(CH 3 )-, -CH 2 CH(CH 3 )-, -CH 2 CH 2 CH (CH 3 )-, -CH 2 CH 2 CH 2 CH(CH 3 )-, -CH 2 CH 2 CH 2 CH 2 CH(CH 3 )-, -CH 2 CH 2 CH 2 CH 2 CH 2 CH(CH 3 ) a combination of one or more of -, -(CH 2 ) i O- and -(CH 2 ) i CO-, i is an integer from 0 to 5; and/or,
    所述X 2为
    Figure PCTCN2018077229-appb-100008
    和/或，     The X 2 is
    Figure PCTCN2018077229-appb-100008
    and / or,
    所述R 3选自：-H、-CH 3、-CH 2CH(CH 3) 2、-CH(CH 3)CH 2CH 3、-CH(CH 3) 2、
    Figure PCTCN2018077229-appb-100009
    -CH 2CH 2SCH 3-、-CH 2OH、-CH 2SH、-CH(OH)CH 3、
    Figure PCTCN2018077229-appb-100010
    -CH 2COOH和-CH 2CH 2COOH；和/或，     The R 3 is selected from the group consisting of: -H, -CH 3 , -CH 2 CH(CH 3 ) 2 , -CH(CH 3 )CH 2 CH 3 , -CH(CH 3 ) 2 ,
    Figure PCTCN2018077229-appb-100009
    -CH 2 CH 2 SCH 3 -, -CH 2 OH, -CH 2 SH, -CH(OH)CH 3 ,
    Figure PCTCN2018077229-appb-100010
    -CH 2 COOH and -CH 2 CH 2 COOH; and/or,
    所述F选自：甲氧基、乙氧基、
    Figure PCTCN2018077229-appb-100011
    The F is selected from the group consisting of: methoxy, ethoxy,
    Figure PCTCN2018077229-appb-100011
    Figure PCTCN2018077229-appb-100012
    Figure PCTCN2018077229-appb-100012
  8. 如权利要求7所述的制备方法，其特征在于，所述X 1为-CH 2CH 2-；和/或，所述X 3为-CH 2-；和/或，所述X 2为-CH 2-或-CH(CH 3)-；和/或，所述F为
    Figure PCTCN2018077229-appb-100013
    The method according to claim 7, wherein X 1 is -CH 2 CH 2 -; and/or X 3 is -CH 2 -; and/or X 2 is - CH 2 - or -CH(CH 3 )-; and/or, the F is
    Figure PCTCN2018077229-appb-100013
    优选地，所述制备方法的反应为：Preferably, the reaction of the preparation method is:
    Figure PCTCN2018077229-appb-100014
    Figure PCTCN2018077229-appb-100014
  9. 一种Y型分支的亲水性聚合物羧酸衍生物，其具有如下结构：A Y-branched hydrophilic polymer carboxylic acid derivative having the following structure:
    Figure PCTCN2018077229-appb-100015
    Figure PCTCN2018077229-appb-100015
    其中，P a和P b为相同或不同的亲水性聚合物残基， Wherein P a and P b are the same or different hydrophilic polymer residues,
    X 1和X 3为连接基团，独立地选自：-(CH 2) i-、
    Figure PCTCN2018077229-appb-100016
    -(CH 2) iO-、-(CH 2) iS-和-(CH 2) iCO-中的一种或多种的组合，i为0-10的整数，     X 1 and X 3 are a linking group independently selected from: -(CH 2 ) i -,
    Figure PCTCN2018077229-appb-100016
    a combination of one or more of -(CH 2 ) i O-, -(CH 2 ) i S- and -(CH 2 ) i CO-, i is an integer from 0 to 10,
    X 2为连接基团，选自：-(CH 2) r-、-(CH 2) rO-、-(CH 2) rS-和
    Figure PCTCN2018077229-appb-100017
    r为0-10的整数，     X 2 is a linking group, chosen from :-( CH 2) r -, - (CH 2) r O -, - (CH 2) r S- , and
    Figure PCTCN2018077229-appb-100017
    r is an integer from 0 to 10,
    R 1和R 2独立地选自：-H、C 1-6取代或未取代的烷基、C 1-6取代或未取代的烷氧基、C 3-6取代或未取代的环烷基和C 4-10取代或未取代的亚烷基环烷基， R 1 and R 2 are independently selected from: -H, C 1-6 substituted or unsubstituted alkyl, C 1-6 substituted or unsubstituted alkoxy, C 3-6 substituted or unsubstituted cycloalkyl And a C 4-10 substituted or unsubstituted alkylene cycloalkyl group,
    R 3选自：-H、C 1-6取代或未取代的烷基、C 6-10取代或未取代的芳烷基和C 4-10取代或未取代的杂环烷基。 R 3 is selected from the group consisting of: -H, a C 1-6 substituted or unsubstituted alkyl group, a C 6-10 substituted or unsubstituted aralkyl group, and a C 4-10 substituted or unsubstituted heterocycloalkyl group.
  10. 如权利要求9所述的羧酸衍生物，其特征在于，所述羧酸衍生物的分子量为15-50KDa；和/或，The carboxylic acid derivative according to claim 9, wherein the carboxylic acid derivative has a molecular weight of 15 to 50 KDa; and/or
    所述的P a选自：聚乙二醇、聚丙二醇、聚乙烯醇、聚四氢呋喃、聚环氧丙烷、聚环氧丁烷、聚氧杂环丁烷和聚丙烯吗啉中的一种或多种的共聚物的残基；和/或， The P a is selected from the group consisting of polyethylene glycol, polypropylene glycol, polyvinyl alcohol, polytetrahydrofuran, polypropylene oxide, polybutylene oxide, polyoxetane and polypropylene morpholine or a residue of a plurality of copolymers; and/or,
    所述的P a的分子量为7.5-25KDa；和/或， The molecular weight of P a is 7.5-25 KDa; and/or,
    所述的P b选自：聚乙二醇、聚丙二醇、聚乙烯醇、聚四氢呋喃、聚环氧丙烷、聚环氧丁烷、聚氧杂环丁烷和聚丙烯吗啉中的一种或多种的共聚物的残基；和/或， The P b is selected from the group consisting of polyethylene glycol, polypropylene glycol, polyvinyl alcohol, polytetrahydrofuran, polypropylene oxide, polybutylene oxide, polyoxetane and polypropylene morpholine or a residue of a plurality of copolymers; and/or,
    所述的P b的分子量为7.5-25KDa。 The P b has a molecular weight of 7.5 to 25 KDa.
  11. 如权利要求10所述的羧酸衍生物，其特征在于，所述的P a为聚乙二醇残基，其结构为R a-O-(CH 2CH 2O) m-，所述R a选自：H、甲基、乙基、异丙基、环丙基、环丁基、环己基和苄基，m为170-565的整数；和/或， The carboxylic acid derivative according to claim 10, wherein said P a is a polyethylene glycol residue having a structure of R a -O-(CH 2 CH 2 O) m -, said R a is selected from the group consisting of: H, methyl, ethyl, isopropyl, cyclopropyl, cyclobutyl, cyclohexyl and benzyl, m is an integer from 170 to 565; and/or,
    所述的P b为聚乙二醇残基，其结构为R b-O-(CH 2CH 2O) n-，R b选自：H、甲基、乙基、异丙基、环丙基、环丁基、环己基和苄基，n为170-565的整数。 The P b is a polyethylene glycol residue having a structure of R b —O—(CH 2 CH 2 O) n —, and R b is selected from the group consisting of: H, methyl, ethyl, isopropyl, cyclopropane Base, cyclobutyl, cyclohexyl and benzyl, n is an integer from 170 to 565.
  12. 如权利要求10所述的羧酸衍生物，其特征在于，所述R a为H或甲基；和/或，所述R b为H或甲基；和/或，所述m与n为相等的整数。 The carboxylic acid derivative according to claim 10, wherein said R a is H or a methyl group; and/or said R b is H or a methyl group; and/or said m and n are An equal integer.
  13. 如权利要求10所述的羧酸衍生物，其特征在于，所述Y型分支的亲水性聚合物羧酸衍生物为Y型分支的聚乙二醇羧酸衍生物，其具有如下结构：The carboxylic acid derivative according to claim 10, wherein the Y-branched hydrophilic polymer carboxylic acid derivative is a Y-branched polyethylene glycol carboxylic acid derivative having the following structure:
    Figure PCTCN2018077229-appb-100018
    Figure PCTCN2018077229-appb-100018
  14. 如权利要求9-13任一项所述的羧酸衍生物，其特征在于，所述X 1和X 3独立地选自：单键、-CH 2-、-CH 2CH 2-、-CH 2CH 2CH 2-、-CH 2CH 2CH 2CH 2-、-CH 2CH 2CH 2CH 2CH 2-、-CH(CH 3)-、-CH 2CH(CH 3)-、-CH 2CH 2CH(CH 3)-、-CH 2CH 2CH 2CH(CH 3)-、-CH 2CH 2CH 2CH 2CH(CH 3)-、-CH 2CH 2CH 2CH 2CH 2CH(CH 3)-、-(CH 2) iO-和-(CH 2) iCO-中的一种或多种的组合，i为0-5的整数；和/或， The carboxylic acid derivative according to any one of claims 9 to 13, wherein X 1 and X 3 are independently selected from the group consisting of: a single bond, -CH 2 -, -CH 2 CH 2 -, -CH 2 CH 2 CH 2 -, -CH 2 CH 2 CH 2 CH 2 -, -CH 2 CH 2 CH 2 CH 2 CH 2 -, -CH(CH 3 )-, -CH 2 CH(CH 3 )-, - CH 2 CH 2 CH(CH 3 )-, -CH 2 CH 2 CH 2 CH(CH 3 )-, -CH 2 CH 2 CH 2 CH 2 CH(CH 3 )-, -CH 2 CH 2 CH 2 CH 2 a combination of one or more of CH 2 CH(CH 3 )-, -(CH 2 ) i O- and -(CH 2 ) i CO-, i is an integer from 0 to 5; and/or,
    所述X 2为
    Figure PCTCN2018077229-appb-100019
    The X 2 is
    Figure PCTCN2018077229-appb-100019
  15. 如权利要求14所述的羧酸衍生物，其特征在于，所述X 1为-CH 2CH 2-；和/或，所述X 3为-CH 2-。 The carboxylic acid derivative according to claim 14, wherein said X 1 is -CH 2 CH 2 -; and/or said X 3 is -CH 2 -.
  16. 如权利要求14所述的羧酸衍生物，其特征在于，所述羧酸衍生物具有如下结构：The carboxylic acid derivative according to claim 14, wherein the carboxylic acid derivative has the following structure:
    Figure PCTCN2018077229-appb-100020
    Figure PCTCN2018077229-appb-100020
    所述R 3选自：-H、-CH 3、-CH 2CH(CH 3) 2、-CH(CH 3)CH 2CH 3、-CH(CH 3) 2、
    Figure PCTCN2018077229-appb-100021
    -CH 2CH 2SCH 3-、-CH 2OH、-CH 2SH、-CH(OH)CH 3、
    Figure PCTCN2018077229-appb-100022
    -CH 2COOH和-CH 2CH 2COOH，优选地，所述R 3为-H或-CH 3。     The R 3 is selected from the group consisting of: -H, -CH 3 , -CH 2 CH(CH 3 ) 2 , -CH(CH 3 )CH 2 CH 3 , -CH(CH 3 ) 2 ,
    Figure PCTCN2018077229-appb-100021
    -CH 2 CH 2 SCH 3 -, -CH 2 OH, -CH 2 SH, -CH(OH)CH 3 ,
    Figure PCTCN2018077229-appb-100022
    -CH 2 COOH and -CH 2 CH 2 COOH, preferably, R 3 is -H or -CH 3 .
  17. 一种基于权利要求9-16任一项所述的羧酸衍生物的Y型分支的亲水性聚合物衍生物，其具有如下结构：A Y-branched hydrophilic polymer derivative of the carboxylic acid derivative according to any one of claims 9 to 16, which has the following structure:
    Figure PCTCN2018077229-appb-100023
    Figure PCTCN2018077229-appb-100023
    其中，X 4为连接基团，选自：-(CH 2) j-、-(CH 2) jO-、-(CH 2) jS-、-(CH 2) jCO-、-(CH 2) jNH-、-(CH 2) jCONH-和-(CH 2) jNHCO-中一种或多种的组合，j为0-10的整数， Wherein X 4 is a linking group selected from the group consisting of: -(CH 2 ) j -, -(CH 2 ) j O-, -(CH 2 ) j S-, -(CH 2 ) j CO-, -(CH 2 ) a combination of one or more of j NH-, -(CH 2 ) j CONH- and -(CH 2 ) j NHCO-, j is an integer from 0 to 10,
    Q为端基，选自：C 1-6的烷氧基、羟基、氨基、羧基、巯基、酯基、酮基、醛基、邻二硫吡啶基、叠氮基、酰肼基、炔基、硅烷基、马来酰亚胺基和琥珀酰亚胺基。 Q is a terminal group selected from: C 1-6 alkoxy, hydroxy, amino, carboxy, thiol, ester, keto, aldehyde, o-dithiopyridyl, azide, hydrazide, alkynyl , silyl, maleimide and succinimide groups.
  18. 如权利要求17所述的衍生物，其特征在于，所述X 4选自：单键、-CH 2-、-CH 2CH 2-、-CH 2CH 2CH 2-、-CH 2CH 2CH 2CH 2-、-CH 2CH 2CH 2CH 2CH 2-、-(CH 2) jCO-、-(CH 2) jNH-、-(CH 2) jCONH-和-(CH 2) jNHCO-中一种或多种的组合，j为0-5的整数；和/或， The derivative according to claim 17, wherein said X 4 is selected from the group consisting of: a single bond, -CH 2 -, -CH 2 CH 2 -, -CH 2 CH 2 CH 2 -, -CH 2 CH 2 CH 2 CH 2 -, -CH 2 CH 2 CH 2 CH 2 CH 2 -, -(CH 2 ) j CO-, -(CH 2 ) j NH-, -(CH 2 ) j CONH- and -(CH 2 j ) a combination of one or more of NHCO-, j being an integer from 0 to 5; and/or,
    所述Q选自：-OH、-SH、-C≡CH、-NH 2、-COOH、-CHO、
    Figure PCTCN2018077229-appb-100024
    和-N 3。     The Q is selected from the group consisting of: -OH, -SH, -C≡CH, -NH 2 , -COOH, -CHO,
    Figure PCTCN2018077229-appb-100024
    And -N 3 .
  19. 如权利要求17所述的衍生物，其特征在于，所述Y型分支的亲水性聚合物衍生物为Y型分支的聚乙二醇衍生物，其具有如下结构：The derivative according to claim 17, wherein the Y-branched hydrophilic polymer derivative is a Y-branched polyethylene glycol derivative having the following structure:
    Figure PCTCN2018077229-appb-100025
    Figure PCTCN2018077229-appb-100025
    或，所述Y型分支的亲水性聚合物衍生物具有如下结构：Alternatively, the Y-branched hydrophilic polymer derivative has the following structure:
    Figure PCTCN2018077229-appb-100026
    Figure PCTCN2018077229-appb-100026
    所述R 3选自：-H、-CH 3、-CH 2CH(CH 3) 2、-CH(CH 3)CH 2CH 3、-CH(CH 3) 2、
    Figure PCTCN2018077229-appb-100027
    -CH 2CH 2SCH 3-、-CH 2OH、-CH 2SH、-CH(OH)CH 3、
    Figure PCTCN2018077229-appb-100028
    -CH 2COOH和-CH 2CH 2COOH；优选地，所述R 3为-H或-CH 3。     The R 3 is selected from the group consisting of: -H, -CH 3 , -CH 2 CH(CH 3 ) 2 , -CH(CH 3 )CH 2 CH 3 , -CH(CH 3 ) 2 ,
    Figure PCTCN2018077229-appb-100027
    -CH 2 CH 2 SCH 3 -, -CH 2 OH, -CH 2 SH, -CH(OH)CH 3 ,
    Figure PCTCN2018077229-appb-100028
    -CH 2 COOH and -CH 2 CH 2 COOH; preferably, the R 3 is -H or -CH 3 .
  20. 一种如权利要求17-19任一项所述的Y型分支的亲水聚合物衍生物的制备方法，其包括如权利要求1-8任一项所述的制备方法步骤。A method of producing a Y-branched hydrophilic polymer derivative according to any one of claims 17 to 19, which comprises the steps of the preparation method according to any one of claims 1-8.
  21. 一种如权利要求9-16任一项所述的Y型分支的亲水性聚合物羧酸衍生物、权利要求17-19任一项所述的Y型分支的亲水聚合物衍生物与药物的结合物。A Y-branched hydrophilic polymer carboxylic acid derivative according to any one of claims 9 to 16, a Y-branched hydrophilic polymer derivative according to any one of claims 17 to 19, and a combination of drugs.
  22. 一种包括权利要求21所述的结合物及任选的药学上可接受的载体或赋形剂的药物组合物。A pharmaceutical composition comprising the combination of claim 21 and, optionally, a pharmaceutically acceptable carrier or excipient.
  23. 一种如权利要求9-16任一项所述的Y型分支的亲水性聚合物羧酸衍生物、权利要求17-19任一项所述的Y型分支的亲水聚合物衍生物、权利要求21所述的结合物或权利要求22所述的药物组合物在制备治疗疾病的药物中的应用。A Y-branched hydrophilic polymer carboxylic acid derivative according to any one of claims 9 to 16, a Y-branched hydrophilic polymer derivative according to any one of claims 17 to 19, Use of the combination of claim 21 or the pharmaceutical composition of claim 22 for the manufacture of a medicament for the treatment of a disease.
PCT/CN2018/077229 2017-03-30 2018-02-26 Method for preparing y-branched hydrophilic polymer carboxylic acid derivative WO2018177055A1 (en)

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CA2304976A1 (en) * 1997-11-05 1999-05-14 J. Milton Harris Delivery of poly(ethylene glycol)-conjugated molecules from degradable hydrogels
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