CN1757620A - Biphase identification chiral prepn. of chromatographic chiral compound(s) - Google Patents

Abstract

This invention discloses a new type of two-phase identified chirality preparation chromatography separation technique which introduces chirality stationary solid phase and chirality eluate phase, it can make the separation process of chirality preparation chromatography more economical and as well the higher purity of the separated products - the enantiomer.

Description

Biphase identification chiral preparative chromatography resolving chiral compound

Technical field:

The present invention relates to a kind of novel disassemble technique of chiral compound enantiomer, belong to separation technology field.

Background technology:

The research of current chipal compounds is most important most active part in stereochemistry, the pharmaceutical chemistry.Because the enantiomer of chipal compounds has different biological activitys usually, the chipal compounds that therefore prepares enantiomer-pure is all significant in fields such as life science, pharmaceutical chemistry, materials chemistries.Because the enantiomorph of chipal compounds is except optical property and the active difference in asymmetric catalysis synthesis, other physics, chemical property are then identical, so it is very difficult to obtain the compound of enantiomer-pure.The method for preparing at present the enantiomer-pure chipal compounds in the world mainly comprises the direct crystallization fractionation of asymmetric synthesis method, racemic modification, chemical fractionation, kinetic resolution and biological fractionation and liquid phantom preparing chromatogram method etc.The outstanding problem that above-mentioned these methods exist is that the scope of application is narrower, there is the enantiomer-pure product of many materials still can not adopt these methods to obtain, and operation is loaded down with trivial details, yield is lower, and the optical purity of product is undesirable, production cost is too high, causes the price of enantiopure compound such as enantiopure pharmaceuticals all high 3～200 times than the racemic modification form.The many enantiomer-pure chiral drugs and the initial chiral raw material of China's use at present rely on import in a large number, and most of chirality synthetic drugss that China produces are still with the form of racemic modification and sell, and the new and effective disassemble technique of therefore developing chipal compounds especially has great importance to the development of China's pharmaceutical industry, pesticide industry.

Liquid chromatography technology is a kind of isolation technique that extensively adopts in the industry such as chemical industry, light industry, food, biology, medicine.In fields such as pharmacy, biotechnology, fine chemistry industry, liquid phantom preparing chromatogram has obtained application more and more widely in recent years.Chromatogram is separated different components, and its principle is based between different components and the chromatographic stationary phase surface in the system to be separated has different absorption properties, and the rate of migration when causing various components to flow with moving phase in chromatographic column is different to realize separation.In recent years the technology that occurs with liquid phantom preparing chromatogram method resolving chiral compound, the surface of this stratographic stationary phase is made of chiral molecules, be called chiral stationary phase, utilize the difference of the adsorptivity of different enantiomorphs on the chiral surfaces of chromatogram chiral stationary phase in the chiral racemic system, rate of migration difference when causing different enantiomorphs in chromatographic column, to flow with moving phase, so the weak enantiomorph of adsorptive power flows out chromatographic column with moving phase earlier, and flow out chromatographic column behind the strong enantiomorph of adsorptive power, thereby the fractionation that realizes different enantiomorphs separates.The technology of liquid phantom preparing chromatogram method resolving chiral compound has the irreplaceable advantage of many other method for splitting. little as energy consumption, but separation purity is high, yield is high and normal-temperature operation etc., but also there is simultaneously too high and the weakness that can not operate continuously of process cost, hindered its further application in industrial production.Based on these pretty conditions, people have invented many improved technology, as cyclical chromatography, continuous chromatography etc., and wherein the most attractive be simulated moving bed chromatography.The later stage nineties, the Belgium scientist takes the lead in simulation moving-bed liquid chromatography is introduced in the split process of chipal compounds, on the basis of optimized choice chiral stationary phase, successfully being moving phase with methyl alcohol has split the U-26225A enantiomorph continuously with the speed of 20.2g/d, has opened up the novel process with the simulation moving-bed liquid chromatography technology resolving chiral of chirality compound.Greatly reduce the consumption and the running cost of solvent.Because but the simulation moving-bed liquid chromatography technology of chirality has the advantage that operate continuously, operational conditions gentleness, solvent and separating medium can be recycled, greatly reduce the consumption and the running cost of solvent, once extensive concern occurring promptly causing, in short so far several years, developed rapidly, fully show the vigorous growth momentum of this technology, become a member new in the liquid phantom preparing chromatogram disassemble technique.At present the problem that exists of liquid phantom preparing chromatogram disassemble technique is: it is little that 1. employed chiral stationary phase itself is treated the difference of the adsorptive power of different enantiomorphs in the racemize system of fractionation, just chiral selectivity is poor, cause the time difference of different enantiomorphs outflow chromatographic columns less, cause the bands of a spectrum of different enantiomorphs to be overlapped and be difficult to effective separation, employed chiral stationary phase is coating-type substantially simultaneously, its selective adsorption to different enantiomorph molecules in the moving phase only depends on one deck chiral molecules of silica gel surface applied, causes the chirality composition on chiral stationary phase surface to be very easy to loss and to cause chromatographic column to lose efficacy; 2. the different enantiomorphs that have only the chiral molecules on chiral stationary phase surface to treat fractionation have selectivity, and there is not the chirality composition in the composition of moving phase, therefore in split process for waiting that splitting system only plays carrier and conveying person, by treating the selective adsorption effect that the suitable solvability that splits system increases chiral stationary phase, in addition to the not more direct effect of selectivity of split process, especially do not see the report that on the basis of chiral stationary phase, adopts the liquid phantom preparing chromatogram disassemble technique of chirality moving phase simultaneously with chirality characteristics.Above-mentioned reason causes the solvent solute of present liquid phantom preparing chromatogram disassemble technique bigger than very (≈ 360), and processing power is little, and enantiomeric purity is not high and to split cost still higher, demands further fully research urgently so that it adapts to real industrial application requirements as early as possible.

Summary of the invention

The purpose of this invention is to provide the liquid phantom preparing chromatogram disassemble technique that different enantiomorphs have better choice in a kind of racemize system for the treatment of fractionation, it can make split process in the liquid phantom preparing chromatogram become economical and to split the enantiomeric purity of product higher.

For realizing purpose of the present invention, method of the present invention also is to be flow through chiral stationary phase and made the different enantiomorphs in the racemize system to be split obtain splitting by the different enantiomorphs that moving phase is carried in the racemize system to be split, it is characterized in that in the moving phase except that containing the achirality composition, also contain the different enantiomorphs chiral component all inequality in the racemize system at least a and to be split and constitute chirality moving phase with chirality characteristics.These chiral component become chirality and help and select agent, be with phasing and different racemize systems to be split and through screening at different chiralitys, except that keeping the suitable solvability of chirality moving phase for racemize system to be split, these chiralitys help selects agent can also be on the chiral stationary phase surface with respect to waiting to split different enantiomorphs in the system in various degree competitive adsorption to take place, thereby further increased the difference that chiral stationary phase is treated the adsorptive power that splits the different enantiomorphs in the system, the time difference that different enantiomorphs flow out chromatographic column is further increased, reach better fractionation effect.Also should note it when the chirality in screening chirality moving phase helps and selects agent and wait to split different enantiomorphs in the system in the difference of boiling point, properties such as water-soluble, so that after fractionation is finished, separate with these enantiomorphs by proper method, recycle with receiving chirality moving phase, obtain the fractionation product of generated in high enantiomeric purity simultaneously.

This liquid phase chirality preparative chromatography disassemble technique of chiral stationary phase and chirality moving phase that adopts simultaneously of the present invention had both had the selective adsorption recognition reaction of chiral stationary phase to different enantiomorphs, also have of the aid identification effect of chirality moving phase simultaneously, so can be referred to as the chirality preparative chromatography disassemble technique of two-phase identification to different enantiomorphs.Adopt biphase identification chiral preparative chromatography technology that racemic modification system is split, compare with the chiral separation technology of independent use chiral stationary phase, the time difference that different enantiomorphs in the racemic modification flow out chromatographic column will further increase, reduce the overlapping of bands of a spectrum, make the sample introduction concentration of racemize sample can be bigger, the processing power of increase equipment, split the employed moving phase of unit product still less, the enantiomeric purity that splits product simultaneously is higher, split process is more economical, reaches better fractionation effect.The aid identification effect of chirality moving phase can greatly improve the efficient of chiral stationary phase in addition, reduce the hop count and the length of chromatographic column, reduce facility investment, simultaneously, select agent by in chirality moving phase, selecting suitable helping, can make same chiral stationary phase can finish fractionation to multiple chirality enantiomorph system, the versatility of the expensive chiral stationary phase of improving price.

Embodiment

Describe method of the present invention and advantage in detail below in conjunction with the specific embodiment that splits ketamine and Proprasylyte enantiomorph.

Embodiment 1: ketamine is surgical clinical a kind of parenteral nerve narcotic commonly used, be used as medicine with the racemic modification form at present always, but pharmacological research shows, the anesthetic action of R (+) isomer is 3 times of S (-)-isomer in the racemize ketamine, and S (-)-ketamine not only anesthetic action is little, and make patient emotional, lose oneself's control easily, even lose the side effect of judgement, this has not only limited the clinical use of racemize ketamine, also making in recent years, the racemize ketamine has the trend that develops into drugs, therefore efficient R (+)-ketamine that the racemize ketamine is split as chiral purity is used as medicine in research, S (-)-ketamine is destroyed in the source that produces, not only can formulate efficient new drug, resist aspect the overflow of drugs also highly significant.We are chiral stationary phase with the beta-cyclodextrin of the aminopropyl silica gel surface bond phenyl isocyanate derivatize of granularity 5 μ m, the 250mm that packs into is long, in the chromatographic column of internal diameter 4.6mm, with V (normal hexane)/V (Virahol)=80/20 is moving phase, flow rate of mobile phase 1.0mL/min, racemize ketamine sample dissolves with moving phase, make the sample solution of 20mg/mL, once instantaneous sample introduction 0.025mL, under described chromatographic condition, split, two Separation of Enantiomers degree are 1.08 only, are unrealized better to split; When add the R-2-amylalcohol in above-mentioned moving phase is that chirality helps and selects agent, after making the chirality moving phase that contains the 4.5mmol/LR-2-amylalcohol, under above-mentioned identical chromatographic condition, split, two Separation of Enantiomers degree reach 2.04, the chirality moving phase that contains in the enantiomorph product can reclaim through the low temperature rotary evaporation, has fully shown the remarkable effect under the two-phase condition for identification;

Embodiment 2: the beta-cyclodextrin with the aminopropyl silica gel surface bond phenyl isocyanate derivatize of granularity 40～60 μ m is a chiral stationary phase, 12 250mm that pack into are long, in the chromatographic column of internal diameter 4.6mm, the combined annular simulated moving bed chromatography, with V (normal hexane)/V (Virahol)=80/20 is moving phase, flow rate of mobile phase 0.6mL/min in the simulated moving bed chromatography, racemize ketamine sample dissolves with moving phase, make the sample solution of 20mg/mL, sample introduction flow velocity 0.1mL/min, split under described chromatographic condition, the enantiomeric purity that obtains two enantiomorph products only is respectively 90.1% and 92.3%, is unrealized better to split; When add the R-2-amylalcohol in above-mentioned moving phase is that chirality helps and selects agent, after making the chirality moving phase that contains the 4.5mmol/LR-2-amylalcohol, under above-mentioned identical chromatographic condition, split, the enantiomeric purity of two enantiomorph products reaches 99.91% and 99.96% respectively, the chirality moving phase that contains in the enantiomorph product has also fully shown the advantage that two-phase identification splits through the recycling use of low temperature rotary evaporation;

Embodiment 3: Proprasylyte is a kind of important beta-blockers, be used as medicine with the form of racemic modification always, experimentation on animals shows, S (-)-Proprasylyte is bigger approximately 100 times than R (+)-Proprasylyte to the retarding degree of beta receptor, and R (+)-Proprasylyte not only the retardation to beta receptor is very little, and contraceptive efficacy arranged, can be used as a kind of effective contraceptive bian, therefore it is not good that the Proprasylyte that is used as medicine with the racemic modification form is not only treated the usefulness of cardiac system disease, and bring side effect easily, so the racemize Proprasylyte be split as chirally purified compound and be used as medicine respectively as two kinds of different medicines, be the new drug that initiative has the own intellecture property of China, improve the effective ways of China's Proprasylyte drug quality.We are chiral stationary phase with the beta-cyclodextrin of the aminopropyl silica gel surface bond phenyl isocyanate derivatize of granularity 5 μ m, the 250mm that packs into is long, in the chromatographic column of internal diameter 4.6mm, with V (normal hexane)/V (Virahol)=80/20 is moving phase, flow rate of mobile phase 0.6mL/min, racemize Proprasylyte sample dissolves with moving phase, make the sample solution of 20mg/mL, once instantaneous sample introduction 0.025mL, under described chromatographic condition, split, two Separation of Enantiomers degree are 1.06 only, are unrealized better to split; When add the R-sec-butylamine in above-mentioned moving phase is that chirality helps and selects agent, after making the chirality moving phase that contains 4.5mmol/L R-sec-butylamine, under above-mentioned identical chromatographic condition, split, two Separation of Enantiomers degree reach 1.82, the chirality moving phase that contains in the enantiomorph product can reclaim through the low temperature rotary evaporation, has fully shown the superiority that two-phase identification chromatogram splits;

Embodiment 4: the Mierocrystalline cellulose with the aminopropyl silica gel surface-coated triphenyl isocyanic ester derivatize of granularity 5 μ m is a chiral stationary phase, the 250mm that packs into is long, in the chromatographic column of internal diameter 4.6mm, with V (normal hexane)/V (ethanol)=85/15 is moving phase, flow rate of mobile phase 0.8mL/min, all the other dissolve with moving phase for the Proprasylyte sample of S (-)-Proprasylyte to contain R (+)-Proprasylyte 40%, make the sample solution of 20mg/mL, once instantaneous sample introduction 0.025mL, under described chromatographic condition, split, two Separation of Enantiomers degree are 1.05 only, are unrealized better to split; For helping, the mixing chirality selects agent when in above-mentioned moving phase, adding R-sec-butyl alcohol and R-sec-butylamine, after making the chirality moving phase that contains 0.5mmol/L R-sec-butyl alcohol and 0.5mmol/L R-sec-butylamine, under above-mentioned identical chromatographic condition, split, two Separation of Enantiomers degree reach 1.32, the chirality moving phase that contains in the enantiomorph product can reclaim through the low temperature rotary evaporation, and the good effect that two-phase identification chromatogram splits is described.

Claims (5)

1, a kind of liquid phantom preparing chromatogram disassemble technique of resolution of racemic system, this method also is to carry different enantiomorphs in the racemize system to be split by moving phase to flow through chiral stationary phase and make the rate of migration of different enantiomorphs on chiral stationary phase in the racemize system to be split different and obtain splitting, it is characterized in that: in the moving phase except that containing the achirality composition, the different enantiomorphs chirality that is called all inequality that also contains in the racemize system at least a and to be split helps the chiral component of selecting agent, constitute chirality moving phase with chirality characteristics, make two or more enantiomorphs in the racemize system to be split help under the effect of selecting agent in chirality, the difference of the rate of migration on chiral stationary phase further increases, thereby easier realization splits.

2, disassemble technique as claimed in claim 1 is characterized in that: described chiral stationary phase both can be that bonding type also can be the chiral stationary phase of application type.

3, as claim 1 or the described disassemble technique of claim 2, it is characterized in that: described racemize system to be split both can be the racemize system that does not have opticity fully, also can be multiple enantiomorph each other optically active isomer but wherein one or more isomer are excessive systems with respect to other optically active isomers.

4, as claim 1 or claim 2 or the described disassemble technique of claim 3, it is characterized in that: described liquid phantom preparing chromatogram both can be the liquid phantom preparing chromatogram of the single injected sampling of periodical operation, also can be the liquid phantom preparing chromatogram of operate continuously, as cyclical chromatography, continuous chromatography, simulated moving bed chromatography etc.

5, as claim 1 or claim 2 or claim 3 or the described disassemble technique of claim 4, it is characterized in that: described racemize system to be split is racemize ketamine or racemize Proprasylyte.