CN102367290B - Chain-functionalized multi-level branched polyethylene glycol and synthesis method thereof - Google Patents
Chain-functionalized multi-level branched polyethylene glycol and synthesis method thereof Download PDFInfo
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- 229920001223 polyethylene glycol Polymers 0.000 title claims abstract description 93
- 239000002202 Polyethylene glycol Substances 0.000 title claims abstract description 82
- 238000001308 synthesis method Methods 0.000 title abstract 2
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 claims abstract description 17
- 125000001495 ethyl group Chemical group [H]C([H])([H])C([H])([H])* 0.000 claims abstract description 16
- 239000003999 initiator Substances 0.000 claims abstract description 10
- 150000001298 alcohols Chemical class 0.000 claims abstract description 8
- 238000006116 polymerization reaction Methods 0.000 claims abstract description 6
- 125000000217 alkyl group Chemical group 0.000 claims abstract description 3
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 claims description 40
- WYURNTSHIVDZCO-UHFFFAOYSA-N Tetrahydrofuran Chemical compound C1CCOC1 WYURNTSHIVDZCO-UHFFFAOYSA-N 0.000 claims description 40
- RTZKZFJDLAIYFH-UHFFFAOYSA-N ether Substances CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 claims description 35
- -1 1-ethoxyethyl Chemical group 0.000 claims description 28
- 238000006243 chemical reaction Methods 0.000 claims description 26
- 238000002360 preparation method Methods 0.000 claims description 20
- YLQBMQCUIZJEEH-UHFFFAOYSA-N tetrahydrofuran Natural products C=1C=COC=1 YLQBMQCUIZJEEH-UHFFFAOYSA-N 0.000 claims description 20
- CTKINSOISVBQLD-UHFFFAOYSA-N Glycidol Chemical compound OCC1CO1 CTKINSOISVBQLD-UHFFFAOYSA-N 0.000 claims description 18
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 18
- IRAPFUAOCHNONS-UHFFFAOYSA-N potassium;phenylmethylbenzene Chemical compound [K+].C=1C=CC=CC=1[CH-]C1=CC=CC=C1 IRAPFUAOCHNONS-UHFFFAOYSA-N 0.000 claims description 13
- 239000002904 solvent Substances 0.000 claims description 9
- 238000001035 drying Methods 0.000 claims description 8
- 230000035484 reaction time Effects 0.000 claims description 8
- 230000008859 change Effects 0.000 claims description 6
- 239000005457 ice water Substances 0.000 claims description 6
- INQOMBQAUSQDDS-UHFFFAOYSA-N iodomethane Chemical compound IC INQOMBQAUSQDDS-UHFFFAOYSA-N 0.000 claims description 6
- 230000000630 rising effect Effects 0.000 claims description 6
- 125000002887 hydroxy group Chemical group [H]O* 0.000 claims description 5
- 238000010189 synthetic method Methods 0.000 claims description 5
- RDHPKYGYEGBMSE-UHFFFAOYSA-N bromoethane Chemical compound CCBr RDHPKYGYEGBMSE-UHFFFAOYSA-N 0.000 claims description 3
- 239000007810 chemical reaction solvent Substances 0.000 claims description 2
- 239000003795 chemical substances by application Substances 0.000 claims description 2
- 229960001760 dimethyl sulfoxide Drugs 0.000 claims 2
- 229920001184 polypeptide Polymers 0.000 abstract description 14
- 102000004196 processed proteins & peptides Human genes 0.000 abstract description 14
- 108090000765 processed proteins & peptides Proteins 0.000 abstract description 14
- 230000002779 inactivation Effects 0.000 abstract description 5
- 230000008878 coupling Effects 0.000 abstract description 4
- 238000010168 coupling process Methods 0.000 abstract description 4
- 238000005859 coupling reaction Methods 0.000 abstract description 4
- 102000004169 proteins and genes Human genes 0.000 abstract 3
- 108090000623 proteins and genes Proteins 0.000 abstract 3
- AOJDZKCUAATBGE-UHFFFAOYSA-N bromomethane Chemical compound Br[CH2] AOJDZKCUAATBGE-UHFFFAOYSA-N 0.000 abstract 1
- CBOIHMRHGLHBPB-UHFFFAOYSA-N hydroxymethyl Chemical group O[CH2] CBOIHMRHGLHBPB-UHFFFAOYSA-N 0.000 abstract 1
- 229910052697 platinum Inorganic materials 0.000 description 28
- BASFCYQUMIYNBI-UHFFFAOYSA-N platinum Substances [Pt] BASFCYQUMIYNBI-UHFFFAOYSA-N 0.000 description 28
- 239000003814 drug Substances 0.000 description 23
- 239000000243 solution Substances 0.000 description 18
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 description 17
- LYCAIKOWRPUZTN-UHFFFAOYSA-N Ethylene glycol Chemical compound OCCO LYCAIKOWRPUZTN-UHFFFAOYSA-N 0.000 description 17
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 16
- KWYUFKZDYYNOTN-UHFFFAOYSA-M Potassium hydroxide Chemical compound [OH-].[K+] KWYUFKZDYYNOTN-UHFFFAOYSA-M 0.000 description 12
- ZMANZCXQSJIPKH-UHFFFAOYSA-N Triethylamine Chemical compound CCN(CC)CC ZMANZCXQSJIPKH-UHFFFAOYSA-N 0.000 description 12
- 238000001556 precipitation Methods 0.000 description 12
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 11
- 238000005481 NMR spectroscopy Methods 0.000 description 11
- 239000004698 Polyethylene Substances 0.000 description 11
- 229910052739 hydrogen Inorganic materials 0.000 description 11
- 239000001257 hydrogen Substances 0.000 description 11
- 229920000573 polyethylene Polymers 0.000 description 11
- 238000001228 spectrum Methods 0.000 description 11
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 9
- 125000000524 functional group Chemical group 0.000 description 9
- 229920003171 Poly (ethylene oxide) Polymers 0.000 description 8
- 229940079593 drug Drugs 0.000 description 8
- WGCNASOHLSPBMP-UHFFFAOYSA-N hydroxyacetaldehyde Natural products OCC=O WGCNASOHLSPBMP-UHFFFAOYSA-N 0.000 description 7
- 238000000034 method Methods 0.000 description 7
- 239000000047 product Substances 0.000 description 7
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 6
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 6
- 230000007059 acute toxicity Effects 0.000 description 6
- 231100000403 acute toxicity Toxicity 0.000 description 6
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 5
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 5
- 238000000605 extraction Methods 0.000 description 5
- 125000005448 ethoxyethyl group Chemical group [H]C([H])([H])C([H])([H])OC([H])([H])C([H])([H])* 0.000 description 4
- 230000007062 hydrolysis Effects 0.000 description 4
- 238000006460 hydrolysis reaction Methods 0.000 description 4
- 230000004048 modification Effects 0.000 description 4
- 238000012986 modification Methods 0.000 description 4
- 239000002244 precipitate Substances 0.000 description 4
- 239000000126 substance Substances 0.000 description 4
- RYHBNJHYFVUHQT-UHFFFAOYSA-N 1,4-Dioxane Chemical compound C1COCCO1 RYHBNJHYFVUHQT-UHFFFAOYSA-N 0.000 description 3
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 description 3
- YXFVVABEGXRONW-UHFFFAOYSA-N Toluene Chemical compound CC1=CC=CC=C1 YXFVVABEGXRONW-UHFFFAOYSA-N 0.000 description 3
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 description 3
- 229960004316 cisplatin Drugs 0.000 description 3
- 238000001727 in vivo Methods 0.000 description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 3
- NMPVEAUIHMEAQP-UHFFFAOYSA-N 2-Bromoacetaldehyde Chemical compound BrCC=O NMPVEAUIHMEAQP-UHFFFAOYSA-N 0.000 description 2
- YYROPELSRYBVMQ-UHFFFAOYSA-N 4-toluenesulfonyl chloride Chemical compound CC1=CC=C(S(Cl)(=O)=O)C=C1 YYROPELSRYBVMQ-UHFFFAOYSA-N 0.000 description 2
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 2
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 2
- QNAYBMKLOCPYGJ-REOHCLBHSA-N L-alanine Chemical compound C[C@H](N)C(O)=O QNAYBMKLOCPYGJ-REOHCLBHSA-N 0.000 description 2
- 241001465754 Metazoa Species 0.000 description 2
- JUJWROOIHBZHMG-UHFFFAOYSA-N Pyridine Chemical compound C1=CC=NC=C1 JUJWROOIHBZHMG-UHFFFAOYSA-N 0.000 description 2
- PXIPVTKHYLBLMZ-UHFFFAOYSA-N Sodium azide Chemical compound [Na+].[N-]=[N+]=[N-] PXIPVTKHYLBLMZ-UHFFFAOYSA-N 0.000 description 2
- 239000002253 acid Substances 0.000 description 2
- 235000004279 alanine Nutrition 0.000 description 2
- 125000003368 amide group Chemical group 0.000 description 2
- 229910002091 carbon monoxide Inorganic materials 0.000 description 2
- 230000003013 cytotoxicity Effects 0.000 description 2
- 231100000135 cytotoxicity Toxicity 0.000 description 2
- 238000004090 dissolution Methods 0.000 description 2
- 238000009826 distribution Methods 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 235000015097 nutrients Nutrition 0.000 description 2
- 229920000642 polymer Polymers 0.000 description 2
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 2
- 230000009467 reduction Effects 0.000 description 2
- SQGYOTSLMSWVJD-UHFFFAOYSA-N silver(1+) nitrate Chemical compound [Ag+].[O-]N(=O)=O SQGYOTSLMSWVJD-UHFFFAOYSA-N 0.000 description 2
- 238000003756 stirring Methods 0.000 description 2
- 230000004083 survival effect Effects 0.000 description 2
- 150000003536 tetrazoles Chemical class 0.000 description 2
- 238000001291 vacuum drying Methods 0.000 description 2
- PUPZLCDOIYMWBV-UHFFFAOYSA-N (+/-)-1,3-Butanediol Chemical compound CC(O)CCO PUPZLCDOIYMWBV-UHFFFAOYSA-N 0.000 description 1
- ZJIFDEVVTPEXDL-UHFFFAOYSA-N (2,5-dioxopyrrolidin-1-yl) hydrogen carbonate Chemical compound OC(=O)ON1C(=O)CCC1=O ZJIFDEVVTPEXDL-UHFFFAOYSA-N 0.000 description 1
- 0 **N(C(C=C1)=O)C1=O Chemical compound **N(C(C=C1)=O)C1=O 0.000 description 1
- 238000005160 1H NMR spectroscopy Methods 0.000 description 1
- LRWZZZWJMFNZIK-UHFFFAOYSA-N 2-chloro-3-methyloxirane Chemical compound CC1OC1Cl LRWZZZWJMFNZIK-UHFFFAOYSA-N 0.000 description 1
- WFDIJRYMOXRFFG-UHFFFAOYSA-N Acetic anhydride Chemical compound CC(=O)OC(C)=O WFDIJRYMOXRFFG-UHFFFAOYSA-N 0.000 description 1
- NLHHRLWOUZZQLW-UHFFFAOYSA-N Acrylonitrile Chemical compound C=CC#N NLHHRLWOUZZQLW-UHFFFAOYSA-N 0.000 description 1
- 208000024893 Acute lymphoblastic leukemia Diseases 0.000 description 1
- 208000014697 Acute lymphocytic leukaemia Diseases 0.000 description 1
- 108010024976 Asparaginase Proteins 0.000 description 1
- 102000015790 Asparaginase Human genes 0.000 description 1
- 206010006187 Breast cancer Diseases 0.000 description 1
- 208000026310 Breast neoplasm Diseases 0.000 description 1
- 239000006145 Eagle's minimal essential medium Substances 0.000 description 1
- OTMSDBZUPAUEDD-UHFFFAOYSA-N Ethane Chemical group CC OTMSDBZUPAUEDD-UHFFFAOYSA-N 0.000 description 1
- VGGSQFUCUMXWEO-UHFFFAOYSA-N Ethene Chemical compound C=C VGGSQFUCUMXWEO-UHFFFAOYSA-N 0.000 description 1
- 239000005977 Ethylene Substances 0.000 description 1
- 108091006006 PEGylated Proteins Proteins 0.000 description 1
- 208000006664 Precursor Cell Lymphoblastic Leukemia-Lymphoma Diseases 0.000 description 1
- OFOBLEOULBTSOW-UHFFFAOYSA-N Propanedioic acid Natural products OC(=O)CC(O)=O OFOBLEOULBTSOW-UHFFFAOYSA-N 0.000 description 1
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 description 1
- 125000003158 alcohol group Chemical group 0.000 description 1
- 230000000118 anti-neoplastic effect Effects 0.000 description 1
- 230000000259 anti-tumor effect Effects 0.000 description 1
- 239000002246 antineoplastic agent Substances 0.000 description 1
- 229940041181 antineoplastic drug Drugs 0.000 description 1
- 229960003272 asparaginase Drugs 0.000 description 1
- DCXYFEDJOCDNAF-UHFFFAOYSA-M asparaginate Chemical compound [O-]C(=O)C(N)CC(N)=O DCXYFEDJOCDNAF-UHFFFAOYSA-M 0.000 description 1
- 229960000074 biopharmaceutical Drugs 0.000 description 1
- 229940040591 biotech drug Drugs 0.000 description 1
- AXNCLTXKOHIWCA-UHFFFAOYSA-N butan-1-ol;propane-1,2-diol Chemical class CCCCO.CC(O)CO AXNCLTXKOHIWCA-UHFFFAOYSA-N 0.000 description 1
- 244000309466 calf Species 0.000 description 1
- 150000001732 carboxylic acid derivatives Chemical class 0.000 description 1
- 239000006285 cell suspension Substances 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- DQLATGHUWYMOKM-UHFFFAOYSA-L cisplatin Chemical compound N[Pt](N)(Cl)Cl DQLATGHUWYMOKM-UHFFFAOYSA-L 0.000 description 1
- 239000012141 concentrate Substances 0.000 description 1
- 231100000433 cytotoxic Toxicity 0.000 description 1
- 230000001472 cytotoxic effect Effects 0.000 description 1
- 239000008367 deionised water Substances 0.000 description 1
- 229910021641 deionized water Inorganic materials 0.000 description 1
- 239000003405 delayed action preparation Substances 0.000 description 1
- MTHSVFCYNBDYFN-UHFFFAOYSA-N diethylene glycol Chemical compound OCCOCCO MTHSVFCYNBDYFN-UHFFFAOYSA-N 0.000 description 1
- 230000029087 digestion Effects 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 239000003937 drug carrier Substances 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 150000002148 esters Chemical class 0.000 description 1
- HCPOCMMGKBZWSJ-UHFFFAOYSA-N ethyl 3-hydrazinyl-3-oxopropanoate Chemical compound CCOC(=O)CC(=O)NN HCPOCMMGKBZWSJ-UHFFFAOYSA-N 0.000 description 1
- 239000000706 filtrate Substances 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 125000002485 formyl group Chemical group [H]C(*)=O 0.000 description 1
- 235000011187 glycerol Nutrition 0.000 description 1
- 150000001261 hydroxy acids Chemical group 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 229910052500 inorganic mineral Inorganic materials 0.000 description 1
- 239000010410 layer Substances 0.000 description 1
- 231100000636 lethal dose Toxicity 0.000 description 1
- 230000031700 light absorption Effects 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 239000012280 lithium aluminium hydride Substances 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 238000001840 matrix-assisted laser desorption--ionisation time-of-flight mass spectrometry Methods 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- 239000012982 microporous membrane Substances 0.000 description 1
- 239000011707 mineral Substances 0.000 description 1
- 235000010755 mineral Nutrition 0.000 description 1
- 229940099216 oncaspar Drugs 0.000 description 1
- 230000001590 oxidative effect Effects 0.000 description 1
- 230000007170 pathology Effects 0.000 description 1
- 108010001564 pegaspargase Proteins 0.000 description 1
- 230000006320 pegylation Effects 0.000 description 1
- 230000002572 peristaltic effect Effects 0.000 description 1
- 230000001376 precipitating effect Effects 0.000 description 1
- YORCIIVHUBAYBQ-UHFFFAOYSA-N propargyl bromide Chemical compound BrCC#C YORCIIVHUBAYBQ-UHFFFAOYSA-N 0.000 description 1
- 239000011241 protective layer Substances 0.000 description 1
- UMJSCPRVCHMLSP-UHFFFAOYSA-N pyridine Natural products COC1=CC=CN=C1 UMJSCPRVCHMLSP-UHFFFAOYSA-N 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 210000002966 serum Anatomy 0.000 description 1
- DTPQZKZONQKKSU-UHFFFAOYSA-N silver azanide silver Chemical compound [NH2-].[Ag].[Ag].[Ag+] DTPQZKZONQKKSU-UHFFFAOYSA-N 0.000 description 1
- 229910001961 silver nitrate Inorganic materials 0.000 description 1
- 229940126586 small molecule drug Drugs 0.000 description 1
- 150000003384 small molecules Chemical class 0.000 description 1
- 239000001632 sodium acetate Substances 0.000 description 1
- 235000017281 sodium acetate Nutrition 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
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- Polyethers (AREA)
- Medicinal Preparation (AREA)
Abstract
The invention discloses a chain-functionalized multi-level branched polyethylene glycol and a synthesis method thereof. The chain-functionalized multi-level branched polyethylene glycol has a molecular weight of 2,000-1,000,000 and has a following structural formula, wherein 1-99% of R1 independently is H; the remaining R1 independently is CH2OH, CH2OCH2CH2CN, CH2OCH2CH2CH2NH2, CH2Br, CH2N3, CH2OCH2CHO, CH2OCH2CH2COOH, CH2OCH2CH2CONH2, CH2OCH2CH=CH2, or CH2OCH2CHis equivalent to CH2, or R is an alkyl of an alcohols initiator; X is a methyl or ethyl; z is the number of X groups; n1, n2 ellipsis are respectively the degrees of polymerization of 1,2 ellipsis level branched chains and are integers from 10 to 500; the branched level is not less than 2; and the number of the branched chains on each level branch is not less than 2. Such a multi-level branched polyethylene glycol has a plurality of functionalized groups capable of being subjected to homologous protein or polypeptide coupling. The protein or polypeptide can be completely wrapped, thereby resisting the attacks of outside macromolecular matters and further reducing the inactivation of the protein or polypeptide in an organism.
Description
Technical field
The present invention relates to a kind of branched polyethylene glycol and preparation method thereof, specifically refer to multi-level branched polyethylene glycol and synthetic method thereof that a kind of chain is functionalized.
Background technology
At present, in the research of bio-pharmaceutical carrier, most study be that hydroxyl or the carboxyl of linear and branching (both arms and multi-arm) polyoxyethylene glycol are modified the functional group that changes into high reaction activity, as active ester, aldehyde radical, amino etc.Be compared to the linear polyethylene glycol of same molecular amount, branching (both arms and the multi-arm) polyoxyethylene glycol that contains simple function group has larger molecular volume, thereby more is conducive to modification and modification to small-molecule drug.Simultaneously; due to special molecular conformation; this branched polyethylene glycol can form on the top layer of medicine the protective layer of one deck umbrella shape; increased sterically hindered around drug molecule; the linear polyethylene glycol that compares is more effective has stoped the attack of other macromolecular complex confrontation medicine in body, has reduced medicine inactivation or by the degree of enzymic hydrolysis in vivo.
Through research during the last ten years, existing more than the 10 kind of Pegylation biotech drug approval that obtains U.S. Food and Drug Administration (FDA) is in the market gone on the market.For example the exploitation of Enzon company with the polyethyleneglycol modified medicine ONCASPAR that obtains to the asparaginase, be used for the treatment of acute lymphoblastic leukemia (Pegylated Protein Drugs:Basic Science and Clinical).
Yet the method also has following two kinds of main shortcomings:
1) because this technology just is connected an end of polyoxyethylene glycol with albumen or polypeptide, can not effectively albumen or polypeptide be isolated fully with the external world, albumen or polypeptide non-inactivation adequately protect;
2) because an end that only has polyoxyethylene glycol has active group and can be connected with albumen or polypeptide, with albumen or polypeptide coupling the time, efficient is extremely low, usually needs the polyoxyethylene glycol of excessive 5～20 times just can react fully.
Summary of the invention
The invention provides the functionalized multi-level branched polyethylene glycol of a kind of chain and synthetic method thereof, in order to solve the above-mentioned problems in the prior art, this kind multi-level branched polyethylene glycol with a plurality of can be with the functionalized group of albumen or polypeptide coupling, albumen or polypeptide can be wrapped up fully, fully extraneous its attack of macromolecular complex confrontation of opposing further reduces albumen or polypeptide inactivation in vivo.
Technical scheme provided by the invention is as follows:
The multi-level branched polyethylene glycol that chain is functionalized, its molecular weight are 2,000～1,000,000, and have following structural formula:
Wherein, 1～99% R
1Be H independently, remaining R
1Be CH independently
2OH or CH
2OCH
2CH
2CN or CH
2OCH
2CH
2CH
2NH
2Or CH
2Br or CH
2N
3Or CH
2OCH
2CHO or CH
2OCH
2CH
2COOH or CH
2OCH
2CH
2CONH
2Or CH
2OCH
2CH=CH
2Or CH
2OCH
2CH ≡ CH
2Or
In the present invention, R can be alcohol compound arbitrarily, can be specifically methyl alcohol, ethanol, ethylene glycol, propylene glycol butanols, butyleneglycol or glycerine etc.
The carrier that the multi-level branched polyethylene glycol of said structure can be used as sustained release preparation is widely used in biomedical pharmacy field, has very excellent industrialization prospect.This kind multi-level branched polyethylene glycol with a plurality of can be with the functionalized group of albumen or polypeptide coupling, albumen or polypeptide can be wrapped up fully, fully extraneous its attack of macromolecular complex confrontation of opposing further reduces albumen or polypeptide inactivation in vivo.
Another object of the present invention is to provide the synthetic method of the functionalized multi-level branched polyethylene glycol of a kind of chain.
The polymerization procedure of alcohols initiator (ROH) and oxyethane, 1-ethoxyethyl group-2,3 epoxy propyl ether is as follows:
A, dimethyl sulfoxide (DMSO) (DMSO) solution of alcohols initiator and diphenyl-methyl potassium (DPMK) is added in the closed reactor of anhydrous and oxygen-free, and reactor is placed in 0 ℃ of ice-water bath, the mol ratio of alcohols initiator and diphenyl-methyl potassium (DPMK) is 1: 1;
B, the oxyethane (EO) that adds calculated amount and 1-ethoxyethyl group-2,3 epoxy propyl ether (EGEE), reacted 24～48 hours rising temperature to 0～50 ℃;
C, (1) under the prerequisite that does not change the reactor stopping property, slowly are added dropwise to dimethyl sulfoxide (DMSO) (DMSO) solution of Racemic glycidol (glycidol) subsequently, temperature of reaction are increased to 100～120 ℃ simultaneously, and the reaction times is 4～12 hours;
(2) add the oxyethane (EO) of calculated amount and 1-ethoxyethyl group-2,3 epoxy propyl ether (EGEE), reacted 24～48 hours rising temperature to 0～50 ℃;
As required, repeating step (1) and (2) are repeatedly;
D, add excessive diphenyl-methyl potassium (DPMK) solution at last, then add excessive monobromethane (Et-Br) or methyl iodide (Me-I), temperature of reaction is at 0～50 ℃, and the reaction times is 12～48 hours; Reactor is opened, after solvent is concentrated, is precipitated in a large amount of 0 ℃ of anhydrous diethyl ether, filter, drying, namely obtain the end hydroxyl by X be methyl or ethyl, functional group is the functionalized multi-level branched polyethylene glycol of chain of hydroxy-protective group.
N wherein
1, n
2, n
3... be respectively 1,2, the polymerization degree of 3...... level branched chain and be 10～500 integer, wherein branch progression be not less than 2 and Branches of Different Orders on branched chain quantity be not less than 2; 1～99%R
1Be H, remaining R
1Be ethoxyethyl group.
To 1～99%R
1For after the multi-level branched polyethylene glycol of ethoxyethyl group modifies, can obtain 1～99%R
1Be CH independently
2OH or CH
2OCH
2CH
2CN or CH
2OCH
2CH
2CH
2NH
2Or CH
2Br or CH
2N
3Or CH
2OCH
2CHO or CH
2OCH
2CH
2COOH or CH
2OCH
2CH
2CONH
2Or CH
2OCH
2CH=CH
2Or CH
2OCH
2CH ≡ CH
2Or
The functionalized multi-level branched polyethylene glycol of chain.
In the present invention, 1～99%R
1Be CH
2The preparation process of the multi-level branched polyethylene glycol of OH is as follows: add the 1～99%R that makes in the round-bottomed flask of dried and clean
1For ethoxyethyl group, X be the multi-level branched polyethylene glycol of methyl or ethyl, as the tetrahydrofuran (THF) of reaction solvent with as the concentrated hydrochloric acid of acidolysis agent, react and be spin-dried for tetrahydrofuran (THF) and concentrated hydrochloric acid after 15 minutes, namely obtain 1～99%R
1Be CH
2The multi-level branched polyethylene glycol of OH.
In the present invention, using 1～99%R
1Be CH
2OH, X are multi-level branched polyethylene glycol and the CH of methyl or ethyl
2=CH-CN reaction preparation CH
2OCH
2CH
2During CN functional group, the solvent that uses is dioxane, and potassium hydroxide is catalyzer, 1～99%R that it generates
1Be CH
2OCH
2CH
2CN, X are that the multi-level branched polyethylene glycol of methyl or ethyl can make 1～99%R in 24 hours in By Hydrolysis At Room Temperature in example hydrochloric acid under the mineral acid condition
1Be CH
2OCH
2CH
2CONH
2, X is the multi-level branched polyethylene glycol of methyl or ethyl, 1～99%R
1Be CH
2OCH
2CH
2CONH
2, X can make 1～99%R in 24 hours in By Hydrolysis At Room Temperature during the multi-level branched polyethylene glycol of methyl or ethyl continues under the inorganic alkaline condition as potassium hydroxide
1Be CH
2OCH
2CH
2COOH, X are the multi-level branched polyethylene glycol of methyl or ethyl; In addition, 1～99%R
1Be CH
2OCH
2CH
2CN, X be methyl or ethyl multi-level branched polyethylene glycol at room temperature lucifuge obtain 1～99%R by lithium aluminium hydride reduction
1Be CH
2OCH
2CH
2CH
2NH
2, X is the multi-level branched polyethylene glycol of methyl or ethyl.
In the present invention, 1～99%R
1Be multi-level branched polyethylene glycol and the Br-CH of methyl or ethyl for hydroxyl, X
2-CH=CH
2, Br-CH
2-C ≡ CH,
Br
2Or
(n=1～5)
Reaction prepares 1～99%R
1Be CH
2OCH
2CH=CH
2, CH
2OCH
2CH ≡ CH
2,
CH
2Br or CH
2OCH
2During the multi-level branched polyethylene glycol of CHO, solvent is tetrahydrofuran (THF) or toluene, and diphenyl-methyl potassium is proton reagent.1～99%R wherein
1Be CH
2The multi-level branched polyethylene glycol of Br can continue reaction and make CH under room temperature and lucifuge in dimethyl formamide with sodiumazide
2N
3
In the present invention, 1～99%R
1For hydroxyl, X be methyl or ethyl multi-level branched polyethylene glycol with
The reaction preparation
During functional group, the solvent that uses is acetonitrile, and triethylamine is catalyzer.
In the present invention, 1～99%R
1Be CH
2OCH
2CH
2CH
2NH
2, X be methyl or ethyl multi-level branched polyethylene glycol with
The reaction preparation
Two steps of functional group's time-division carry out: the first step: 1～99%R
1Be NH
2Multi-level branched polyethylene glycol make
Open loop generates amido linkage and a hydroxy-acid group, and this step is take methylene dichloride as solvent, and temperature of reaction is at 70～85 ℃, and the reaction times is 0.5～2 hour; Second step: amido linkage and carboxylic acid loop production
Functional group, this step, sodium acetate was catalyzer take diacetyl oxide as solvent, and temperature of reaction is at 90～110 ℃, and the reaction times is 0.5～2 hour.
The functionalized multi-level branched polyethylene glycol of the chain for preparing in the present invention can instead precipitate by just precipitating, and the methods such as extraction are carried out repeatedly purifying, and all have very high yield.
The functionalized multi-level branched polyethylene glycol of the chain for preparing in the present invention has narrower molecular weight distribution, and structure is clear and definite, can pass through MALDI-TOF MS, SEC or
1H NMR means characterize in detail.
Preparation method's preparation time cycle of the present invention is short, and chain length, chain number and functional number can accurately be controlled by the charging capacity of regulating ring oxidative ethane, 1-ethoxyethyl group-2,3 epoxy propyl ether and Racemic glycidol, with having very excellent industrialization prospect.
Embodiment
The present invention will be further described below in conjunction with embodiment, but do not consist of any limitation of the invention.
Embodiment 1
The successive polymerization of oxyethane, 1-ethoxyethyl group-2,3 epoxy propyl ether and Racemic glycidol:
Will (R-OH, R be CH as the ethanol of initiator
3CH
2) and dimethyl sulfoxide (DMSO) (DMSO) solution of diphenyl-methyl potassium (DPMK) be added in the closed reactor of anhydrous and oxygen-free, and reactor is placed in 0 ℃ of ice-water bath, the mol ratio of initiator and diphenyl-methyl potassium is 1: 1.
Adding molar weight is the oxyethane (EO) of 4: 1 and 1-ethoxyethyl group-2,3 epoxy propyl ether, and reacted 24～48 hours rising temperature to 0～50 ℃.
(1) subsequently under the prerequisite that does not change the reactor stopping property, slowly be added dropwise to dimethyl sulfoxide (DMSO) (DMSO) solution of Racemic glycidol (glycidol) with peristaltic pump, temperature of reaction is increased to 100～120 ℃ simultaneously, the reaction times is 4～12 hours.
(2) adding molar weight is the oxyethane (EO) of 4: 1 and 1-ethoxyethyl group-2,3 epoxy propyl ether, and reacted 24～48 hours rising temperature to 0～50 ℃.
Repeating step (1) and step (2) 1 times.
Add at last excessive diphenyl-methyl potassium (DPMK) solution, then add excessive methyl iodide (Me-I), temperature of reaction is at 0～50 ℃, and the reaction times is 12～48 hours.Reactor is opened, after solvent is concentrated, precipitated in a large amount of 0 ℃ of anhydrous diethyl ether, filter, drying namely obtains 20%R
1Be ethoxyethyl group, 80%R
1Be three grades of branched polyethylene glycols of methyl for H, X.
This wherein, n
1, n
2And n
3The hydrogen spectrum that is three grades of branched polyethylene glycols of 50 is as follows:
1H?NMR(CDCl
3)δ(ppm)：1.20(CH
3CH
2-)，1.35(-OCH(CH
3)O-)，3.35(CH
3O-)，3.40-3.80(-CH
2CH
2O-，CH
3CH
2-，glycidol)，4.75(-OCH(CH
3)O-)。
Embodiment 2
20%R
1Be CH
2OH, X are CH
3The preparation of three grades of branched polyethylene glycols:
Polymkeric substance, 20mL tetrahydrofuran (THF) and the 0.5mL concentrated hydrochloric acid of the methyl iodide end-blocking that makes add 1g embodiment 1 in the 50mL of dried and clean round-bottomed flask in react and are spin-dried for tetrahydrofuran (THF) and concentrated hydrochloric acid after 15 minutes, namely obtain 20%R
1Be CH
2OH, X are CH
3Multi-level branched polyethylene glycol.20%R
1For-CH
2OH, X are CH
3The hydrogen spectrum data of multi-level branched polyethylene glycol as follows:
1H?NMR(CDCl
3)δ(ppm)：3.35(CH
3O-)，3.40-3.80(-CH
2CH
2O-，glycidol)。
Embodiment 3
20%R
1Be CH
2OCH
2CH
2CN, X are CH
3The preparation of three grades of branched polyethylene glycols:
The 20%R that makes add 1g embodiment 2 in the 50mL of dried and clean round-bottomed flask in
1Be CH
2OH, X are CH
3Multi-level branched polyethylene glycol, 20mL dioxane and 0.2g potassium hydroxide, slowly drip the 1mL vinyl cyanide in the time of 0 ℃.At room temperature continue reaction after 8 hours, revolve to steam and remove dioxane, with product be dissolved in wash three times in chloroform with water after, chloroform is used anhydrous magnesium sulfate drying mutually, filter, concentrated, precipitation obtains 20%R
1Be CH
2OCH
2CH
2CN, X are CH
3Multi-level branched polyethylene glycol.20%R
1Be CH
2OCH
2CH
2CN, X are CH
3The hydrogen spectrum data of multi-level branched polyethylene glycol as follows:
1H?NMR(CDCl
3)δ(ppm)：2.40(-CH
2CN)，3.35(CH
3O-)，3.40-3.80(-CH
2CH
2O-，glycidol)。
Embodiment 4
20%R
1Be CH
2OCH
2CH
2CONH
2, X is CH
3The preparation of three grades of branched polyethylene glycols:
The 20%R that makes add 1g embodiment 3 in the 50mL of dried and clean round-bottomed flask in
1Be CH
2OCH
2CH
2CN, X are CH
3Multi-level branched polyethylene glycol and 15mL concentrated hydrochloric acid, after at room temperature reacting 48 hours, add entry dilution, and with twice of chloroform extraction.Chloroform is used anhydrous magnesium sulfate drying mutually, filter, concentrated, precipitation obtains 20%R
1Be CH
2OCH
2CH
2CONH
2, X is CH
3Multi-level branched polyethylene glycol.20%R
1Be CH
2OCH
2CH
2CONH
2, X is CH
3The hydrogen spectrum data of multi-level branched polyethylene glycol as follows:
1H?NMR(CDCl
3)δ(ppm)：2.55(-CH
2CONH
2)，3.35(CH
3O-)，3.40-3.80(-CH
2CH
2O-，glycidol)。
Embodiment 5
20%R
1Be CH
2OCH
2CH
2COOH, X are CH
3The preparation of three grades of branched polyethylene glycols:
The 20%R that makes add 1g embodiment 4 in the 50mL of dried and clean round-bottomed flask in
1Be CH
2OCH
2CH
2CONH
2, X is CH
3Multi-level branched polyethylene glycol, 30mL water and 2g potassium hydroxide.After at room temperature reacting 24 hours, and with twice of chloroform extraction.Chloroform is used anhydrous magnesium sulfate drying mutually, filter, concentrated, precipitation obtains 20%R
1Be CH
2OCH
2CH
2COOH, X are CH
3Multi-level branched polyethylene glycol.20%R
1Be CH
2OCH
2CH
2COOH, X are CH
3The hydrogen spectrum data of multi-level branched polyethylene glycol as follows:
1H?NMR(CDCl
3)δ(ppm)：2.60(-CH
2COOH)，3.35(CH
3O-)，3.40-3.80(-CH
2CH
2O-，glycidol)。
Embodiment 6
20%R
1Be CH
2OCH
2CH ≡ CH
2, X is CH
3The preparation of three grades of branched polyethylene glycols:
The 20%R that makes add 1g embodiment 2 in the 50mL of dried and clean round-bottomed flask in
1Be CH
2OH, X are CH
3Methyl-sulphoxide (DMSO) solution of multi-level branched polyethylene glycol, 20mL tetrahydrofuran (THF) and 2mL diphenyl-methyl potassium (DPMK), after at room temperature reacting 2 hours, slowly add the 2.0mL propargyl bromide and continue reaction 12 hours.Concentrated afterwards, precipitation obtains 20%R
1Be CH
2OCH
2CH ≡ CH
2, X is CH
3Multi-level branched polyethylene glycol.20%R
1Be CH
2OCH
2CH ≡ CH
2, X is CH
3The hydrogen spectrum data of multi-level branched polyethylene glycol as follows:
1H?NMR(CDCl
3)δ(ppm)：3.35(CH
3O-)，3.40-3.80(-CH
2CH
2O-，glycidol)，4.05(-OCH
2CΞCH)。Embodiment 7
20%R
1Be CH
2OCH
2CHO, X are CH
3The preparation of three grades of branched polyethylene glycols:
The 20%R that makes add 1g embodiment 2 in the 50mL of dried and clean round-bottomed flask in
1Be CH
2OH, X are CH
3Methyl-sulphoxide (DMSO) solution of multi-level branched polyethylene glycol, 20mL tetrahydrofuran (THF) and 2mL diphenyl-methyl potassium (DPMK), after at room temperature reacting 2 hours, slowly add 2.0mL bromoacetaldehyde diglycol ethylene and continue reaction 12 hours.Concentrated afterwards, precipitation, and precipitated product was dissolved in again in the hydrochloric acid soln of 30mL 10% reaction 12 hours, use afterwards twice of chloroform extraction.Chloroform is used anhydrous magnesium sulfate drying mutually, filter, concentrated, precipitation obtains 20%R
1Be CH
2OCH
2CHO, X are CH
3Multi-level branched polyethylene glycol.20%R
1Be CH
2OCH
2CHO, X are CH
3The hydrogen spectrum data of multi-level branched polyethylene glycol as follows:
1H?NMR(CDCl
3)δ(ppm)：3.35(CH
3O-)，3.40-3.80(-CH
2CH
2O-，glycidol)，4.65(-OCH
2CHO)。
Embodiment 8
20%R
1For
X is CH
3The preparation of three grades of branched polyethylene glycols:
The 20%R that makes add 1g embodiment 2 in the 50mL of dried and clean round-bottomed flask in
1Be CH
2OH, X are CH
3Multi-level branched polyethylene glycol, 20mL acetonitrile, 4mL triethylamine and 1.0g N, N '-two succinimidyl carbonate, concentrated after at room temperature reacting 24 hours, precipitation obtains 20%R
1For
X is CH
3Multi-level branched polyethylene glycol.20%R
1For
X is CH
3The hydrogen spectrum data of multi-level branched polyethylene glycol as follows:
1H?NMR(CDCl
3)δ(ppm)：2.80(-(O＝)CCH
2CH
2C(＝O)-)，3.35(CH
3O-)，3.40-3.80(-CH
2CH
2O-，glycidol)，4.15(-CH
2OCO-)。
Embodiment 9
20%R
1For
X is CH
3The preparation of three grades of branched polyethylene glycols:
The 20%R that makes add 1g embodiment 2 in the 50mL of dried and clean round-bottomed flask in
1Be CH
2OH, X are CH
3Multi-level branched polyethylene glycol, 20mL pyridine and 0.5g Tosyl chloride, concentrated after at room temperature reacting 24 hours, precipitation obtains 20%R
1For
X is CH
3Multi-level branched polyethylene glycol.20%R
1For
X is CH
3The hydrogen spectrum data of multi-level branched polyethylene glycol as follows:
1H?NMR(CDCl
3)δ(ppm)：2.42(CH
3C
6H
48O
2-)，3.35(CH
3O-)，3.40-3.80(-CH
2CH
2O-，glycidol)，7.50-7.90(CH
3C
6H
4SO
2-)。
Embodiment 10
20%R
1For
X is CH
3The preparation of three grades of branched polyethylene glycols:
The 20%R that makes add 1g embodiment 2 in the 50mL of dried and clean round-bottomed flask in
1Be CH
2OH, X are CH
3The dimethyl sulfoxide solution of multi-level branched polyethylene glycol, 20mL tetrahydrofuran (THF) and 2mL diphenyl-methyl potassium, after at room temperature reacting 2 hours, slowly add the 2.0mL epoxy chloropropane and continue reaction 12 hours.Concentrated afterwards, precipitation namely obtains 20%R
1For
X is CH
3Multi-level branched polyethylene glycol.20%R
1For
X is CH
3The hydrogen spectrum data of multi-level branched polyethylene glycol as follows:
1H?NMR(CDCl
3)δ(ppm)：2.50-2.90(-CH
2OCH-)，3.35(CH
3O-)，3.40-3.80(-CH
2CH
2O-，glycidol，-OCH
2CH＜)。
The present invention also comprises the method for the binding substances of preparation this multi-level branched polyethylene glycol and antitumor drug cis-platinum, and the method comprises, uses above-mentionedly can to react with active group multi-level branched polyethylene glycol and the cis-platinum of two amino reactions on cis-platinum with various.
In order to obtain multi-level branched polyethylene glycol-cis-platinum binding substances, we have adopted functional group is that R is CH
2OCH
2CH
2COOH, further with alanine diethyl ester (DEAM) reaction, through hydrolysis obtain can with the multi-level branched polyethylene glycol of cis-platinum reaction; Last and cis-platinum reaction obtains the antineoplastic cisplatin medicine of multi-level branched polyethylene glycol.
The method that connects cis-platinum about linear polyethylene glycol is existing the description in the art, such as Yuichi etc., and Macromol.Biosci.2001,1, No.8,355-363.
Test example
The preparation of multi-level branched polyethylene glycol-cis-platinum
Take 42g alanine diethyl ester (DEAM) (20mmol), be dissolved in the 60mL anhydrous methylene chloride, add triethylamine 12mL, stirred 3 hours under ice-water bath.With the carboxyl multi-level branched polyethylene glycol (molecular weight 20 that makes in 10.0g embodiment 5,000, functional group 10 and molecular weight 40,000, functional group 10) be dissolved in the 70mL anhydrous methylene chloride, drip triethylamine and regulate pH to 8～9 (solution is dropped to moistening accurate pH test paper to be recorded).Solution with DEAM under ice-water bath slowly is added drop-wise in the solution of PEG.Remain on and react under ice-water bath after 12 hours, filter, concentrate, and precipitate in anhydrous diethyl ether.The product that obtains is dissolved in the methanol solution of sodium hydroxide (methyl alcohol 90mL, 1mol/L aqueous sodium hydroxide solution 13mL), stirs 2 hours.Salt acid for adjusting pH to 6 with 1mol/L is spin-dried for solvent, adds methylene dichloride again to dissolve, and adds anhydrous magnesium sulfate drying.Filter, concentrated, precipitate in anhydrous diethyl ether.Use methylene dichloride and anhydrous diethyl ether dissolution precipitation twice, vacuum-drying obtains product 8.6g, productive rate 86% again.Its hydrogen spectrum data are as follows:
1H?NMR(CDCl
3)δ(ppm)：3.35(CH
3O-)，3.40-3.80(-CH
2CH
2O-，glycidol)，5.10(-NH-CH(COOH))。
Cis-platinum 1.0g (3.3mmol) is dissolved in the 200mL deionized water, is heated to 60 ℃ and makes it to dissolve fully.Add again Silver Nitrate 1.5g (9mmol), filter the silver nitride precipitation of generation after 6 hours in 60 ℃ of lucifuge stirring reactions.Add 1.0g multi-level branched polyethylene glycol propanedioic acid in filtrate, continue 60 ℃ of lucifuge reactions 24 hours, use dichloromethane extraction product three times, merging is concentrated, precipitates in anhydrous diethyl ether.Use methylene dichloride/anhydrous diethyl ether dissolution precipitation twice, vacuum-drying obtains product multi-level branched polyethylene glycol-cis-platinum again.Productive rate 79%.Through aas determination, in product, the molar content of cis-platinum is respectively 87.1%, 83.8%.
The preparation of linear polyethylene glycol-cis-platinum is according to document Macromol.Biosci.2001,1, No.8, and 355-363, adopting the molecular weight of polyoxyethylene glycol is 2,000 and 4,000, the molar content of cis-platinum is respectively 832%, 84.3%.
Cytotoxic test
With the strain of C6 human breast cancer cell with the DMEM nutrient solution (Dulbecco ' s modified Eagle ' s minimum essential medium) that contains 10% foetal calf serum at 37 ℃, 5%CO
2Cultivate under condition, go down to posterity.Eugonic C6 cell after 0.25% tryptic digestion, is made into the cell suspension that final concentration is 70000 cell/mL, this suspension is evenly added in the 96 flat culture plates in hole by 100 μ L/ holes.Every porocyte number is 7000.At 37 ℃, 5%CO
2, hatched under the condition of humidity 100% 24 hours.Above-mentioned each macromolecular drug is dissolved in the DMEM nutrient solution, uses the 0.22um filtering with microporous membrane.The content of gained drug solution cis-platinum in the macromolecular drug is diluted to different concns as benchmark, is converted to the small molecules cis-platinum and is respectively 0.02ug/mL, 0.2ug/mL, 2ug/mL, 20ug/mL, 200ug/mL, 2mg/mL.Add the medicine 100uL/ hole of different concns in the above-mentioned 96 flat culture plates in hole, cultivated 2 days, change liquid, cultivated one day.Every hole adds 20uL 0.5% tetrazole (MTT) solution, and after 4 hours, the sucking-off supernatant liquor adds 150uL dimethyl sulfoxide (DMSO) (DMSO), vibration 10min.Detect the light absorption value (OD value) at 570nm place, each hole with microplate reader, record result, calculate cell survival rate by following formula:
Cell survival rate=experimental group OD value/control group OD value * 100%
The IC of multi-level branched polyethylene glycol-cis-platinum and linear polyethylene glycol-cis-platinum medicine series
50Value
As can be seen from the table, in the homologous series medicine, with respect to the cis-platinum of unmodified, the cytotoxicity of the medicine of polyethyleneglycol modified mistake has all reduced.But the IC of multi-level branched polyethylene glycol-cis-platinum medicine series
50Value is but far below the linear polyethylene glycol of same molecular amount-cis-platinum medicine series.This multi-level branched polyethylene glycol-cisplatin medicine that same molecular amount has been described has embodied better anti tumor activity in vitro than linear polyethylene glycol-cisplatin medicine.
The test of acute toxicity
In order to check the acute toxicity of cis-platinum after PEO modifies to change, measured the LD of above-mentioned polymer drug
50The TA1 small white mouse is divided into 5 groups at random, and 20 every group, male and female half and half are used respectively cis-platinum, have connect the cis-platinum administration of line style and multi-level branched polyethylene glycol, with ip * 1 treatment plan, establishes five metering groups, observes immediate reaction after administration.The death distribution of record mouse in two weeks.Take 24 hours as peak mortality.The dead animal postmortem is not found macroscopic, obvious substantial pathology.After two weeks, the surviving animals generalized case is good.Experimental result is calculated with the Bliss method.
The LD of multi-level branched polyethylene glycol-cis-platinum and linear polyethylene glycol-cis-platinum medicine series
50Value
The medium lethal dose that can see all polymeric medicines in table is all much higher than the cis-platinum of unmodified, is 60 times of cis-platinum at least.This shows, after the modification of polyoxyethylene glycol, the acute toxicity of cis-platinum obviously reduces, and the reduction of polymeric medicine acute toxicity can make up cytotoxicity fully and reduce the loss that brings.In addition, we can not find out that polymer architecture (multi-level branched or line style) on the impact of acute toxicity, can only see that polymericular weight is on the impact of its generation from table.For the polymkeric substance of same structure, molecular weight is larger, and acute toxicity is less.
Above-described embodiment is the better embodiment of the present invention; but embodiments of the present invention are not restricted to the described embodiments; other any do not deviate from change, the modification done under spirit of the present invention and principle, substitutes, combination, simplify; all should be the substitute mode of equivalence, within being included in protection scope of the present invention.
Claims (4)
1. the functionalized multi-level branched polyethylene glycol of chain, its molecular weight is 2,000 ~ 1,000,000, and has following general structure:
Or
Or
······
Wherein, 1～99% R
1Be H independently, remaining R1 is CH independently
2OH or CH
2OCH
2CH
2CN or CH
2OCH
2CH
2CH
2NH
2Or CH
2Br or CH
2N
3Or CH
2OCH
2CHO or CH
2OCH
2CH
2COOH or CH
2OCH
2CH
2CONH
2Or CH
2OCH
2CH=CH
2Or CH
2OCH
2CH ≡ CH or
Or
R is the alkyl of alcohols initiator, and X is methyl or ethyl, and z is the number of X group, n
1, n
2, n
3Be respectively the polymerization degree of 1,2,3 grade of branched chain and be 10～500 integer, wherein branch progression be not less than 2 and Branches of Different Orders on branched chain quantity be not less than 2.
2. the functionalized multi-level branched polyethylene glycol of chain described according to claim 1, is characterized in that: described R
1Be CH
2OH.
3. the synthetic method of the functionalized multi-level branched polyethylene glycol of the chain of claim 2, is characterized in that, comprises the following steps:
A, the dimethyl sulphoxide solution of alcohols initiator and diphenyl-methyl potassium is added in the closed reactor of anhydrous and oxygen-free, and reactor is placed in 0 ℃ of ice-water bath, the mol ratio of alcohols initiator and diphenyl-methyl potassium is 1:1;
B, the oxyethane that adds calculated amount and 1-ethoxyethyl group-2,3 epoxy propyl ether, reacted 24～48 hours rising temperature to 0～50 ℃;
C, ⑴ under the prerequisite that does not change the reactor stopping property, slowly are added dropwise to the dimethyl sulphoxide solution of Racemic glycidol subsequently, temperature of reaction are increased to 100～120 ℃ simultaneously, and the reaction times is 4～12 hours;
⑵ add the oxyethane of calculated amount and 1-ethoxyethyl group-2,3 epoxy propyl ether, and reacted 24～48 hours rising temperature to 0～50 ℃;
As required, repeating step ⑴ and ⑵ are many times;
D, add excessive diphenyl-methyl potassium solution at last, then add excessive monobromethane or methyl iodide, temperature of reaction is at 0～50 ℃, and the reaction times is 12～48 hours; Reactor is opened, after solvent is concentrated, is precipitated in a large amount of 0 ℃ of anhydrous diethyl ether, filter, drying, namely obtain the end hydroxyl by X be methyl or ethyl, 1～99%R
1Be H, all the other 1～99%R
1Be C
2H
5O-CH (CH
3)-O-CH
2-the functionalized multi-level branched polyethylene glycol of chain;
......
E, add the multi-level branched polyethylene glycol that makes in step D in the container of dried and clean, as the tetrahydrofuran (THF) of reaction solvent with as the concentrated hydrochloric acid of acidolysis agent, react and be spin-dried for tetrahydrofuran (THF) and concentrated hydrochloric acid after 15 minutes, namely obtain R
1Be CH
2The multi-level branched polyethylene glycol of OH.
4. the preparation method of the functionalized multi-level branched polyethylene glycol of the chain described according to claim 3, is characterized in that: R
1OH on group can be continued to modify, and obtains R
1Be CH
2OCH
2CH
2CN or CH
2OCH
2CH
2CH
2NH
2Or CH
2Br or CH
2N
3Or CH
2OCH
2CHO or CH
2OCH
2CH
2COOH or CH
2OCH
2CH
2CONH
2Or CH
2OCH
2CH=CH
2Or CH
2OCH
2CH ≡ CH or
Or
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