Summary of the invention
The technical problem to be solved in the present invention is to provide the novel polyoxyethylene glycol aldehyde radical actives of a class.It is simple that such aldehyde radical actives has preparation, the feature of highly versatile.
For solving the problems of the technologies described above, polyoxyethylene glycol aldehyde radical actives of the present invention, has following structure:
Wherein, P is various polyethylene glycols residues, and Z and F are spacer group, and wherein, Z can be selected from: alkyl, amino alkyl residue, the amino residue replacing or be zero (not existing); T is the aldehyde radical number in actives, and t is integer, and scope is: 1≤t≤16; Be preferably 1≤t≤8, more preferably 1≤t≤6, are especially preferably 1≤t≤4.
F in general formula I is selected from: C
bh
calkyl; Wherein, b, c are integer: 0≤b≤20,0≤c≤40; Be preferably 0≤b≤15,0≤c≤30; More preferably 0≤b≤10,0≤c≤20; Especially be preferably 0≤b≤8,0≤c≤16.
P in general formula I comprises: the polyoxyethylene glycol residue of linear chain structure; The polyoxyethylene glycol residue of multi-arm structure; The product residue that carboxylated straight chain polyoxyethylene glycol and the amino-acid residue with polyamino are connected; Wherein, in the polyoxyethylene glycol residue of the polyoxyethylene glycol of linear chain structure and multi-arm structure, have at least the residue of a hydroxyl and Z to be connected: carboxylated straight chain polyoxyethylene glycol and the amino-acid residue with polyamino, this residue is connected with Z by the carboxyl on amino acid.
Wherein, the polyethylene glycols residue of linear chain structure, comprising:
The polyoxyethylene glycol of corresponding these linear chain structure, can adopt commerical prod or can be obtained by simple aggregation, conventionally uses oxyethane, with the method acquisition of ionic polymerization.
The polyethylene glycols residue of multi-arm structure, comprising:
(1) 3 arm polyethylene glycols residue, comprising:
(2) 4 arm polyethylene glycols residues, comprising:
(3) 6 arm polyethylene glycols residues, comprising:
(4) 8 arm polyethylene glycols residues, comprising:
(5) Y type 2 arm polyethylene glycols residues, comprising:
The polyoxyethylene glycol of corresponding these multi-arm structures, can adopt commerical prod, or take polyvalent alcohol as initiator, can be obtained by simple aggregation, conventionally uses oxyethane, with the method acquisition of ionic polymerization.
As a general scheme, the P described in the present invention is not confined to above-mentioned polyoxyethylene glycol residue, and it can be also the product residue that multiple straight chain polyoxyethylene glycol and the amino-acid residue with polyamino are connected, and structure comprises:
Wherein, L is the abbreviation with the amino-acid residue of polyamino, and its general formula is II,
C
vH
wO
uN
y II
V, w, u, y are all integer, 3≤v≤10,6≤w≤20,2≤u≤6,2≤y≤6; Preferably 4≤v≤8,8≤v≤16,2≤y≤4,2≤u≤4; Especially preferably L is lysine residue, and structure is as follows:
But commonly 2 carboxylated straight chain polyoxyethylene glycol and the structure that has two amino amino-acid residues and be connected, be connected it with diamine, obtains a new Y type polyoxamide, and this Y type polyoxamide also can adopt commerical prod.Modal such polyoxamide is to be connected by 2 straight chain polyoxyethylene glycol and lysine residue, then with the quadrol gained that is connected, structure illustrates as follows:
X in P-structure in general formula I of the present invention is number of repeat unit, is also the polymerization degree, and x is integer, 20≤x≤4000; Be preferably 20≤x≤3000; Be more preferably 20≤x≤2000; Especially be preferably 40≤x≤2000.
In the time of the polyoxyethylene glycol residue of the P polyoxyethylene glycol that is linear chain structure or multi-arm structure, Z is C
dh
ealkyl or do not exist, wherein, 1≤d≤10,2≤e≤20; Be preferably 1≤d≤8,2≤e≤16; Be more preferably 1≤d≤6,1≤e≤12; Especially be preferably 1≤d≤4,2≤e≤8;
In the time of the P product residue that to be carboxylated straight chain polyoxyethylene glycol be connected with the amino-acid residue with polyamino, Z is the residue of the amino alkyl replacing or is exactly amino residue, amino on Z is connected with the carboxyl residue of L, Z structure as described in general formula III, wherein, i and h are integer, 0≤i≤10,0≤h≤20, preferably 0≤i≤6,0≤h≤12.
HN——C
iH
h III
For preparation said structure polyoxyethylene glycol aldehyde radical actives of the present invention; when concrete enforcement; usually can adopt commercial polyoxyethylene glycol-alkylamino substituent (again referred to as polyoxamide) that can obtain; carboxylic acid with aldehyde radical protection; under the effect of condensing agent, there is condensation reaction, obtain the protected polyethylene active thing of aldehyde radical, then remove the blocking group of aldehyde radical; obtain target product, take product as straight chain dialdehyde base priming reaction equation schematically as follows:
As adopt the polyoxamide of four arm configurations, structure is as follows:
Can obtain following product:
Because the guard method of aldehyde radical is varied; so the carboxylic acid of aldehyde radical protection is not to only have the acetal protection of above formula a kind of in the present invention; any whenever possible after generation condensation reaction; through deprotection, then the method that obtains the aldehyde radical polyethylene active thing shown in general formula I is all applicable to the present invention.
It should be noted that: because polymer is mutually mixed in together, can not effectively separate, so can not obtain pure polyethylene active thing, polyoxyethylene glycol aldehyde radical actives is no exception.Take 4 arm polyoxamides as example, conventionally by 4 arm polyoxyethylene glycol, through amino-alkylation, reaction obtains, but the transformation efficiency that can not guarantee amino-alkylation reaction is 100%, it is actual that what obtain is a mixture, 1,2,3,4 substitution products and unreacted 4 arm polyoxyethylene glycol are comprised, therefore, in routine duties, can weigh by aldehyde end group replacement rate the quality of product.
Same reason, can not obtain pure 4 arm polyoxyethylene glycol aldehyde radical activess, and only getable be a mixture, comprised 1,2,3,4 substitution products and unreacted 4 arm polyoxamides.Even the methoxy poly (ethylene glycol) aldehyde radical actives for following structure:
Also must obtain the mixture that has comprised methoxy poly (ethylene glycol) amine.
A PEGlike coating aldehyde radical actives of the present invention, the polyethylene glycol structures that comprises straight chain and various multi-arms, therefore, the invention provides the versatility scheme of the polytype polyoxyethylene glycol aldehyde radical actives of preparation, and polyoxyethylene glycol aldehyde radical actives preparation method of the present invention is simple, highly versatile.
The purposes of polyoxyethylene glycol aldehyde radical actives of the present invention is very extensive, as can be used as the modification of bio-active substance, for the Improvement of material, medicine and other fields, provides foundation.As polyoxyethylene glycol aldehyde radical actives of the present invention can be for modifications such as protein, polypeptide, enzyme, somatomedins, by being connected and reaching the object of modification with the terminal amino group of these bio-active substances, as polyoxyethylene glycol aldehyde radical actives can be under acidic conditions, be connected with the amino selectivity of the end of protein matter, concrete grammar can referenced patent: US 5985265A.
Embodiment
Embodiment 1
The methoxy poly (ethylene glycol) amine (5g, molecular weight is 5000) of 0.001mol, puts into the round-bottomed flask of 250ml in the lump with 100ml toluene, heats up toluene distillation to go out, and removes moisture.Add the anhydrous methylene chloride of 100ml, making material dissolution complete, then adding dry 9,9-diethoxy n-nonanoic acid 0.0012mol, adding condensing agent carbonyl dimidazoles 0.0015mol, after 0-5 ℃ of reaction 12h, the insoluble object of elimination, by extremely about 15ml of filtrate concentrated by rotary evaporation, concentrated solution is poured in the Virahol of 150ml, can obtain white solid product, then drying treatment, obtain final product 4.1g, productive rate 82%.
Embodiment 2
Gained 4.1g product in embodiment 1 is dissolved in the phosphate aqueous solution of 4mol/L of 40ml, at 0-5 ℃, stir 10h, divide 3 extractions with 300ml methylene dichloride, by extremely about 15ml of filtrate concentrated by rotary evaporation, concentrated solution is poured in the Virahol of 150ml, can obtain white solid product, then drying treatment, obtain final product 3.6g, productive rate 87%.
NMR (CDCl
3) δ: 3.50 (wide unimodal, the hydrogen in PEG), CH
3o-and-CH
2nHCO-, 3.31~3.40, one unimodal and one three split peak, 5H;-CH
2cH
2nHCO-, 1.72~1.82, five split peak, 2H;-NHCO-, 6.22~6.33, unimodal, 1H;-CH
2cONH-, 2.10~2.17, three split peak, 2H;-CH
2cHO, 2.38~2.45, three split peak, 2H;-CH
2cH
2cHO and-CH
2cH
2cONH-, 1.55~1.68, wide unimodal, 4H; Near its excess-three in aldehyde molecule segment-CH
2-, 1.24~1.38, wide unimodal, 6H;-CHO, 9.74~9.77, unimodal, 1H.On nuclear-magnetism, judge that aldehyde end group replacement rate is greater than 95%.
Embodiment 3
The 4 arm polyoxamides (20g, molecular weight is 20000) of 0.001mol, put into the round-bottomed flask of 500ml in the lump with 200ml toluene, heat up toluene distillation to go out, and remove moisture.Add the anhydrous methylene chloride of 200ml, making material dissolution complete, then adding dry 2,2-diethoxy acetic acid acid 0.0012mol, adding condensing agent carbonyl dimidazoles 0.0015mol, after 0-5 ℃ of reaction 12h, the insoluble object of elimination, by extremely about 45ml of filtrate concentrated by rotary evaporation, concentrated solution is poured in the Virahol of 300ml, can obtain white solid product, then drying treatment, obtain final product 17g, productive rate 85%.
Embodiment 4
Embodiment 3 gained 17g products are dissolved in the phosphate aqueous solution of 4mol/L of 120ml, at 0-5 ℃, stir 10h, divide 3 extractions with 600ml methylene dichloride, by extremely about 35ml of filtrate concentrated by rotary evaporation, concentrated solution is poured in the Virahol of 300ml, can obtain white solid product, then drying treatment, obtain final product 16g, productive rate 94%.
NMR (CDCl
3) δ: 3.50 (wide unimodal, the hydrogen in PEG), C-(CH
2o)
4-and-CH
2nHCO-, 3.31~3.45, one unimodal and one three split peak ,-CHO, 9.74~9.77, unimodal, 1H.On nuclear-magnetism, judge that aldehyde end group replacement rate is greater than 95%.
Embodiment 5
The Y type polyoxamide (10g, molecular weight is 10000) of 0.001mol, structure is as follows:
Put in the lump the round-bottomed flask of 250ml with 100ml toluene, heat up toluene distillation to go out, remove moisture.Add the anhydrous methylene chloride of 100ml, making material dissolution complete, then adding dry 9,9-diethoxy n-nonanoic acid 0.0012mol, adding condensing agent carbonyl dimidazoles 0.0015mol, after 0-5 ℃ of reaction 12h, the insoluble object of elimination, by extremely about 30ml of filtrate concentrated by rotary evaporation, concentrated solution is poured in the Virahol of 250ml, can obtain white solid product, then drying treatment, obtain final product 8.6g, productive rate 86%.
Reaction formula is as follows:
Embodiment 6
Embodiment 5 gained 8.6g products are dissolved in the phosphate aqueous solution of 4mol/L of 80ml, at 0-5 ℃, stir 10h, divide 3 extractions with 800ml methylene dichloride, by extremely about 35ml of filtrate concentrated by rotary evaporation, concentrated solution is poured in the Virahol of 300ml, can obtain white solid product, then drying treatment, obtain final product 8.1g, productive rate 94%.
NMR (CDCl
3) δ: 3.50 (wide unimodal, the hydrogen in PEG and acid amides side CH
2on 2H), C-(CH
2o)
4-and-CH
2nHCO-, 3.31~3.45, one unimodal and one three split peak ,-CHO, 9.74~9.77, unimodal, 1H;-CH
2-CH
2-(amino acid), triplet, 1.56,4H; 6.02, triplet, 1H; 2.9~3.1, amino connected CH
2, 6H.On nuclear-magnetism, judge that aldehyde end group replacement rate is greater than 92%.
Embodiment 7
The Y type polyoxamide (60g, molecular weight is 60000) of 0.001mol, structure is as follows:
Put in the lump the round-bottomed flask of 1000ml with 600ml toluene, heat up toluene distillation to go out, remove moisture.Add the anhydrous methylene chloride of 500ml, making material dissolution complete, then adding dry 9,9-diethoxy n-nonanoic acid 0.0012mol, adding condensing agent carbonyl dimidazoles 0.0015mol, after 0-5 ℃ of reaction 12h, the insoluble object of elimination, by extremely about 200ml of filtrate concentrated by rotary evaporation, concentrated solution is poured in the Virahol of 2500ml, can obtain white solid product, then drying treatment, obtain final product 54g, productive rate 90%.
Reaction formula is as follows:
Embodiment 8
Embodiment 7 gained 54g products are dissolved in the phosphate aqueous solution of 4mol/L of 500ml, at 0-5 ℃, stir 10h, divide 3 extractions with 4000ml methylene dichloride, by extremely about 250ml of filtrate concentrated by rotary evaporation, concentrated solution is poured in the Virahol of 3000ml, can obtain white solid product, then drying treatment, obtain final product 51g, productive rate 94%.
NMR (CDCl
3) δ: 3.50 (wide unimodal, the hydrogen in PEG), C-(CH
2o)
4-and the CH on aldehyde radical side
2, 3.2~3.45, multiplet, 5H;-CHO, 9.74~9.77, unimodal, 1H; CH
2, 2.9, quartet, 2H; CH
2, 1.7, triplet, 2H; .On nuclear-magnetism, judge that aldehyde end group replacement rate is greater than 94%.
Embodiment 9
The methoxy poly (ethylene glycol) amine (2g, molecular weight is 2000) of 0.001mol, structure is as follows:
Put in the lump the round-bottomed flask of 100ml with 200ml toluene, heat up toluene distillation to go out, remove moisture.Add the anhydrous methylene chloride of 50ml, making material dissolution complete, then adding dry 9,9-diethoxy n-nonanoic acid 0.0012mol, adding condensing agent carbonyl dimidazoles 0.0015mol, after 0-5 ℃ of reaction 12h, the insoluble object of elimination, by extremely about 15ml of filtrate concentrated by rotary evaporation, concentrated solution is poured in the Virahol of 200ml, can obtain white solid product, then drying treatment, obtain final product 1.64g, productive rate 82%.
Reaction formula is as follows:
Embodiment 10
Embodiment 9 gained 1.64g products are dissolved in the phosphate aqueous solution of 4mol/L of 20ml, at 0-5 ℃, stir 10h, divide 3 extractions with 200ml methylene dichloride, by extremely about 15ml of filtrate concentrated by rotary evaporation, concentrated solution is poured in the Virahol of 150ml, can obtain white solid product, then drying treatment, obtain final product 1.4g, productive rate 85%.
NMR (CDCl
3) δ: 3.50 (wide unimodal, the hydrogen in PEG), CH
3-(OCH
2cH
2)
4-and the CH2 on aldehyde radical side, 3.2~3.45, multiplet, 5H;-CHO, 9.74~9.77, unimodal, 1H; CH
2, 2.9, quartet, 2H; CH
2, 1.7, triplet, 2H; .On nuclear-magnetism, judge that aldehyde end group replacement rate is greater than 95%.