Background technology
Phosphorylcholine is the terminal hydrophyllic group for constituting the soft phospholipid of membrane structure unit, is the outer layer sense in extracellular tunic
Group, simultaneous with positive and negative xenogenesis electric charge, the ability with stronger combination water and hydrophilicity, the table of this structure and composition
Face is interacted with physiological environment and will not only be adsorbed and depositing proteins, will not also be caused platelet activation, be caused blood coagulation etc.
Untoward reaction, with good biocompatibility.Research in recent years shows, using Phosphorylcholine group and its polymer in material
Material surface construction has anti-cell outer-layer membrane structure, can reduce the absorption of Fibrinogen, and by reducing hematoblastic sticking
Increase the biocompatibility of matrix material with activation.
Therefore in recent years, in order to improve the various functions such as the biocompatibility for improving material, water-retaining property, water absorption, people
Just energetically exploitation imported can with various substrate surfaces reacted it is reactive group, contain phosphinylidyne as dressing agent
The compound of the similar group of choline.For example, patent documentation 1 (CN201310106363.9) discloses a kind of phosphinylidyne gallbladder of hydroxyl
Alkali L- α-glyceryl phosphoryl choline crystal-form compound, patent documentation 2 (CN200580040742.6) disclose a kind of carboxylic phosphorus
The compound of phatidylcholine, patent documentation 3 (CN201310251000.4) disclose a kind of Phosphorylcholine compound containing aldehyde radical.
Though the compound disclosed in above-mentioned patent documentation has imported various functions group, entering with substrate surface treatment agent
During row chemical bonding, under existing technical conditions, must also there is the functional group being complementary in substrate surface, if desired for presence
Amino.If these compounds do not have the help of amino, its response rate can be very low, it is therefore desirable to the reaction of long-time and high temperature
Condition, or grafting process can not be carried out.
Therefore patent documentation 4 (CN201180062050.7) disclose a kind of Phosphorylcholine compound containing mono amino and
Its preparation method, but this kind of compound application still has limitation.This kind of compound only has mono amino functional group, is only capable of conduct
Surface conditioning agent is used for the grafting of substrate surface, and reactivity is relatively low, it is impossible to presence steady in a long-term, additionally, due to mono amino
This kind of compound of limitation cannot function as chain extender or cross-linking agent, so as to can not from material body improve bioaffinity.
The content of the invention
In order to overcome above-mentioned deficiency, the present invention provides a kind of new Phosphorylcholine compound can be directly as the table of material
Face inorganic agent, and as the chain extender and cross-linking agent of material, makes the material containing Phosphorylcholine disguise oneself as the natural component of surrounding,
Can reduce the absorption of Fibrinogen, and hematoblastic stick and activate the bio-compatible that increases matrix material by reducing
Property.
To achieve these goals, the present invention is adopted the following technical scheme that:
A kind of 1B is the Phosphorylcholine compound containing double amino (referred to as Lys-PC) of initiation material synthesis,
This new compound end-blocking with double amino, the Phosphorylcholine group which contains in addition, can make material disguise oneself as it is natural into
Point, its structural formula is as follows:
Here the synthesis of the Phosphorylcholine compound of double amino, from the 1B in gal4 amino acid as starting
Raw material, 1B compare D-Lys and other aminoacid containing polyamino, and its raw material is easy to get, and cost is lower;L- relies in addition
Propylhomoserin contains the aminoacid of double amino, such as agedoite, glutamine, citrulline etc. relative to other, its water-soluble and has
More preferably, structure is simpler for machine thing dissolubility, and in other polyamino aminoacid, second group containing amino is mostly acyl
Amido, amide groups activity are less than amino more, it is difficult to passing through chemical method is changed into amino by amide groups, therefore they are not
The optimum selection of this kind of pair of amino Phosphorylcholine compound of synthesis.
Present invention also offers a kind of preparation method of the Phosphorylcholine compound containing double amino, including:
With 1B as raw material, two amino on lysine are protected, obtain the compound (1) of carboxy blocking;
Under conditions of catalyst is present, the compound (1) of carboxy blocking is reduced into into hydroxy-end capped compound (2);
Compound (2) and 2- oxo -1,3,2- dioxaphospholane (COP) is made to react at low temperature, the chemical combination for obtaining
Thing (3);
Compound (3) is carried out into ring-opening reaction, hydrogenolysis and takes off amido protecting group, obtain final product the Phosphorylcholine containing double amino
Compound.
Preferably, the amido protecting group is Cbz groups, tertbutyloxycarbonyl (Boc), p-toluenesulfonyl (Tosyl)
Or fluorenylmethyloxycarbonyl (Fmoc).
Preferably, the catalyst is N-methylmorpholine.
Synthetic route is as follows:
1. with 1B as initiation material, the aqueous sodium carbonate using 10%-20% as solvent, preferred chloro-carbonic acid benzyl
Ester (Cbz-Cl) protective agent is protected to two amino on lysine, generates N, the double benzyloxycarbonyl group -1Bs of N'-.This
In protect amino functional group also have tertbutyloxycarbonyl (Boc), p-toluenesulfonyl (Tosyl), fluorenylmethyloxycarbonyl (Fmoc) etc.;
The target compound for obtaining, carries out extraction from organic solvents such as dichloromethane, chloroform, dimethyl sulfoxide and can obtain pure
The product of Du Genggao:
Double benzyloxycarbonyl group-the 1Bs of 2.N, N'- are dissolved in tetrahydrofuran or acetonitrile, preferred sodium borohydride (NaBH4) conduct
Reducing agent, with the help of catalyst (the preferred N-methylmorpholine of catalyst), reaction temperature is controlled at-10-- 20 DEG C, by carboxyl
The compound (1) of end-blocking is reduced into hydroxy-end capped compound (2), and which can carry out purification by known method, such as uses successively
Dilute hydrochloric acid, saturated aqueous common salt, distilled water wash choose the Methods For Purification of organic solvent column chromatography:
3. the hydroxy-end capped compound (2) for being obtained with second step is dissolved in anhydrous tetrahydro furan, by 2- oxo -1,3,2- bis-
Oxygen phospholane (COP) is dropped to wherein, and compound (2) is controlled 1 with the amount rate of charge of the material of COP:1~1:1.9;Protect
- 10 DEG C of--25 DEG C of reaction 1-3h of temperature are held, by-product triethylamine hydrochloride is filtered, the compound for obtaining (3) can pass through static,
Recrystallized from acetonitrile, the method purification of acetone recrystallization:
4. the product obtained in step 3 is dissolved in into acetonitrile, -20 DEG C of keeping temperature in reaction bulb rapidly join trimethylamine,
Ring-opening reaction 16-24h is carried out at 50-70 DEG C, then hydrogenolysis is taken off Cbz groups and final product compound (Lys-PC) obtained containing double ammonia
The Phosphorylcholine compound of base, hydrogenolysis method hydrogen bromide (HBr) used acidolysis, palladium charcoal (Pd/C) catalytic hydrogenation (palladium carbon content
5% -20%) etc., here as the reaction of hydrogen bromide acidolysis has by-product to produce, final product has dyeing, therefore we select
The method of catalytic hydrogenation.
Present invention also offers the Phosphorylcholine compound containing double amino prepared by arbitrary above-mentioned method.
Present invention also offers a kind of surface conditioning agent, including:The above-mentioned Phosphorylcholine containing double amino of any one
Compound.
Present invention also offers a kind of chain extender or cross-linking agent, including:The above-mentioned phosphinylidyne gallbladder containing double amino of any one
Alkali cpd.
Present invention also offers the above-mentioned Phosphorylcholine compound containing double amino of any one, surface conditioning agent, chain extension
Connect or cross-linking agent is preparing cosmetics, and organizational project, medicine controlled releasing, the application in gene therapy.
Beneficial effects of the present invention
(1) new compound of the invention is the Phosphorylcholine compound with double amino structures, is that application is high
Compound.Itself can serve as surface conditioning agent, can be as the dressing agent reacted with various substrate surfaces, due to containing
There are double amino, thus improve the Activity and stabill of reaction, dramatically play on material surface and adapted to organism
The surface modified function of the Phosphorylcholine base to represent.For example suppress protein absorption and reduce the soil resistances such as blood coagulation.
(2) using the superiority of the double amino of new compound of the invention, with reference to amino response characteristic (such as nucleophilic displacement of fluorine
Reaction, additive reaction, conjugation substitution reaction etc.), as cross-linking agent or chain extender, and then can obtain various new matrix material
Material, itself improves biocompatibility from material, Phosphorylcholine group is existed long-term and stably, farthest play phosphorus
The function of phatidylcholine group.
(3) the preparation method is that using the 1B of nonhazardouss as initiation material, it is ensured that the compound
Nonhazardouss in medical material are applied to, the lysine in human body also will not produce infringement to human body even if material generation is degraded.
(4) reaction raw materials of the present invention are easy to get, and quantitatively carry out, and high income, step are although more, but purification step is easy.
Embodiment 1
Step 1, the Cbz-Cl of 0.155mol are dissolved in the diethyl ether solution of 100mL, are added dropwise to 200mL containing 0.0645mol
The 10%NaCO of 1B3Aqueous solution, drips 0 DEG C of rear keeping temperature reaction 5h, and the product for obtaining is with appropriate 10% Fructus Citri Limoniae
Chloroform extraction is used after acid acidifying, is evaporated after reaction layer washing and is obtained a large amount of N, the double benzyloxycarbonyl group -1Bs of N'- are produced
Rate about 95%.
Step 2,0.06mol N are taken, the double benzyloxycarbonyl group -1Bs of N'- and 8mL N-methylmorpholines pour 500mL into anti-
In answering bottle, and 200mL tetrahydrofurans are dissolved in, reaction temperature is down to after -15 DEG C the chloromethyl isobutyl ester for adding 0.08mol, stirring
0.38mol sodium borohydrides, the Deca distilled water in reaction bulb is added until no bubble is produced, to continue stirring, room after 15min
The lower reaction 2h of temperature;Take out reaction solution and use dilute hydrochloric acid successively, saturated aqueous common salt, distilled water wash are finally dry with anhydrous magnesium sulfate
It is dry, boil off after solvent obtains crude product, purified with petrol ether/ethyl acetate column chromatography, purity is 96%, yield about 61.5%.
Step 3, product 0.389mol obtained by previous step is taken, poured in 500mL there-necked flasks, with the anhydrous tetrahydrochysene furans of 250mL
Mutter dissolving, after adding 0.389moL triethylamines, keep -15 DEG C of reaction temperature, under mechanical agitation, use constant pressure funnel Deca
The 50mL tetrahydrofuran solutions of 0.389mol COP, after dripping, are further continued for reacting 1.5h in 0.5h, and reduce pressure sucking filtration, is evaporated
To glassy yellow oil sticky mass.Yield about 82%.
Step 4, the product in heatproof, pressure reaction bulb in addition 0.05mol steps 3, the dry second eyeballs of 60mL and magneton,
Put it in freezer compartment of refrigerator, trimethylamine (0.075mol) is added thereto to rapidly after making reactant liquor be cooled to -20 DEG C, suitably
Excessive, sealing is placed, and reacting liquid temperature is recovered to room temperature.Under magnetic agitation, it is placed in 60 DEG C of oil baths and reacts.After reaction 24h
Yellow solution is poured out, is cooled down as refrigerator lower floor and slowly separate out crystallization, crude product dry acetonitrile recrystallization, yield about 91.9%.
Step 5,0.05mol compounds (4) are dissolved in 250mL absolute methanols, after stirring 15min, add under hydrogen shield
Excessive 15%Pd/C, after under room temperature, reaction terminates, solids removed by filtration obtains filtrate, and rotary evaporation removes solvent, uses dry acetonitrile
Recrystallization, obtains final product compound (1) Lys-PC yields about 87%.1H NMR (DMSO, TMS, 400MHz, ppm) δ:5.15
(4H, NH2—);4.13-4.38 (t, 2H ,-CH2OP—);3.13 (s, 9H ,-N+(CH3)3);2.65 (br, 3H,
NH2CH2- ,-CH (NH2)—);1.25-1.55 (br, 6H ,-CH2CH2CH2—)。
Finally it should be noted that the foregoing is only the preferred embodiments of the present invention, this is not limited to
Bright, although being described in detail to the present invention with reference to the foregoing embodiments, for a person skilled in the art, which is still
Technical scheme described in previous embodiment can be modified, or equivalent is carried out to which part.It is all at this
Within bright spirit and principle, any modification, equivalent substitution and improvements made etc. should be included in protection scope of the present invention
Within.Although the above-mentioned specific embodiment to the present invention is described, not limiting the scope of the invention, institute
Category art personnel should be understood that on the basis of technical scheme those skilled in the art need not pay wound
The various modifications made by the property made work or deformation are still within protection scope of the present invention.