A kind of preparation method of chloramphenicol
Technical field
The present invention relates to a kind of preparation method of chloramphenicol, belongs to the technical field of bio-pharmaceuticals and biochemical industry.
Background technology
Chloramphenicol is most one of broad-spectrum antibiotic medicine in the market, and its antibacterial action mechanism is and ribosome 50S
Subunit is combined, and suppresses peptidyl transferase, and so as to suppress protein to synthesize, its structural formula is as follows:
Chloramphenicol is to most of Gram-negatives and positive bacteria, Bacillus influenzae, Bordetella pertussis, shigella dysenteriae, big
Enterobacteria, pneumobacillus, proteus, Pseudomonas aeruginosa, Richettsia, chlamydia trachomatis etc. can effective controls.Due to its
Biomedicine field application widely, therefore explore a kind of gentle, efficiently, economic method prepare chloramphenicol receive it is vast
The concern of chemical-biological worker.
At present the method for industrially prepared chloramphenicol is:With acetophenone as initiation material, by bromination, ammonification, acetylation, hydroxyl
Prepared by the 8 step chemical reactions such as aldehyde condensation, aluminium isopropoxide reduction, hydrolysis deprotection, chiral resolution and dichloroacetyl.Its conjunction
It is as follows into route:
The method is suffered from the drawback that:
(1) this route needs chiral resolution, and the theoretical yield for splitting only has 50%, which results in the yield of whole piece route
Below 30%;In addition, the step of fractionation is very loaded down with trivial details, and to industrial production very big inconvenience is brought;
(2) need to use acetyl group protection, then deprotection in this route, Atom economy is poor and increasings of reactions steps
Plus overall yield is reduced, while also increased difficulty to industrialized production.
(3) aluminium isopropoxide used in this route reduction step, this process produces the three wastes for being difficult to process in a large number, and environment is endangered
Evil is very big.
Therefore, urgently propose that a kind of preparation condition is gentle, operating procedure is simple, product yield effect, Atom economy at present
Superior, environment amenable chloramphenicol preparation method.
The content of the invention
The technical problem to be solved be the technique for preparing chloramphenicol of the prior art exist reactions steps length,
Complex operation step, severe reaction conditions, Atom economy are low, product yield is low, to the disagreeableness problem of environment, and then provide
A kind of inexpensive, easily prepared, yield it is high, more environmentally friendly prepare chloramphenicol method.
In order to solve above-mentioned technical problem, the invention provides the method for preparing chloramphenicol, comprises the steps:With existing
The intermediate (to nitro-alpha-aminoacetophenone hydrochloride) of production of chloramphenicol technique is that initiation material passes through dichloroacetyl, hydroxyl
Aldehyde condensation reaction and asymmetric biocatalytic reduction three-step reaction, obtain final product the chloramphenicol.
It should be noted that the structural formula of the chloramphenicol is as follows:
Preferably, the synthetic route of the chloramphenicol is:
Preferably, the dichloroacetylization reaction is specially:Will to nitro-alpha-aminoacetophenone hydrochloride and solvent, two
Chloracetyl chloride is well mixed, then is added thereto to acid binding agent, reacts at a temperature of 15-20 DEG C, and reaction is finished, and obtains final product two chloroethenes
Acylate.
It should be noted that the reaction dissolvent includes but is not limited to dichloromethane, as long as dissolution can be played i.e.
Can.The acid binding agent includes but is not limited to triethylamine, as long as the sour effect produced in absorbing reaction, ability can be played
Field technique personnel can be adjusted according to the actual requirements to its composition.
It is further preferred that the dichloroacetylization reaction also includes, and in the dichloroacetylization after completion of the reaction, will be anti-
Answer mixed solution to filter, then filter cake is washed with dichloromethane, collect filtrate, and filtrate is used successively watery hydrochloric acid, unsaturated carbonate
Hydrogen sodium solution, saturated common salt water washing, are dried precipitation, obtain final product dichloroacetyl product.
Preferably, the aldol reaction is specially:Dichloroacetyl product is well mixed with ethanol, formaldehyde, is adjusted
The pH value of section mixed solution is 7.2-7.5, is reacted at temperature 32-35 DEG C, and reaction is finished, obtained final product to nitro-α-dichloroacetyl
Amido-beta-hydroxyphenyl acetone.
Preferably, the catalytic reduction reaction is specially:By nitro-α-dichloroacetyl amido-beta-hydroxyphenyl acetone and ketone
Reductase is added in buffer solution, at temperature 20-40 DEG C, pH value be to react under conditions of 6.0-8.0, reaction is finished and obtains final product institute
State chloramphenicol.
Preferably, the cushioning liquid is the PBS cushioning liquid that pH value is 6.0-7.5, concentration is 0.05-0.1mol/L.
It should be noted that the cushioning liquid includes but is not limited to PBS cushioning liquid, as long as can be in enzymic catalytic reaction
When, play and keep salt balance, the cushioning effect of the appropriate pH of adjustment.The PBS cushioning liquid refers to that phosphate-buffered is molten
Liquid, its composition includes but is not limited to Na2HPO4、KH2PO4, NaCl, KCl, those skilled in the art according to the actual requirements can be to it
Composition is adjusted.
Preferably, it is additionally added NADP in the buffer solution+And it is optionally added isopropanol, GDH, grass
One or more in acidohydrogenase, lactic dehydrogenase.
Preferably, it is described to nitro-alpha-aminoacetophenone hydrochloride with nitro-acetophenone as raw material, pass sequentially through bromination anti-
Answer, aminating reaction is obtained.
The present invention has the advantage that compared with prior art:
(1) preparation method of chloramphenicol of the present invention, with the intermediate of existing production of chloramphenicol technique (to nitro-
Alpha-aminoacetophenone hydrochloride) pass through dichloroacetyl, aldol reaction and asymmetric living things catalysis also for initiation material
Former three-step reaction, finally can be up to 84% total recovery,>99% Ee,>99/1 Dr obtains chloramphenicol.Not only avoid
Split and avoid protection-deprotection steps, simplify the production technology of chloramphenicol, improve total recovery, reduce and produce into
This, solve prepare in prior art chloramphenicol the technique for preparing chloramphenicol exist reactions steps length, Atom economy it is low, produce
The low problem of product yield;
(2) preparation method of chloramphenicol of the present invention, simple to operate, mild condition, and the post processing of reaction is simple, easily
Operation, the quantity of three wastes of generation is fewer, environmentally friendly, and complex operation, the pollution for solving other preparation methods of chloramphenicol is big
The shortcomings of, it is adapted to large-scale industrial production.
Specific embodiment
With reference to embodiments, the present invention is further described in detail, but is not limited to this.It should be noted that with
The intermediate (to nitro-alpha-aminoacetophenone hydrochloride) of existing production of chloramphenicol technique is that initiation material passes through dichloroacetyl
Change, aldol condensation and asymmetric biocatalytic reduction three-step reaction, prepare the chloramphenicol.Its synthetic route is:
Embodiment 1-6 is described in detail respectively to dichloroacetyl reaction, aldol reaction and catalytic reduction reaction.
The dichloroacetylization of embodiment 1 is reacted
The present embodiment is the dichloroacetylization reaction in the method for prepare the chloramphenicol, specific as follows:
21.6g is mixed to nitro-alpha-aminoacetophenone hydrochloride with 150ml dichloromethane, 14.5g dichloroacetyl chlorides
It is even, then 21g triethylamine solutions are added thereto to, to react at a temperature of 20 DEG C, reaction is finished, and obtains final product dichloroacetyl product.
Used as the preferred embodiment of the present embodiment, the dichloroacetylization reaction also includes, in the dichloroacetyl
After completion of the reaction, reaction mixture is filtered, then filter cake is washed with dichloromethane, collect filtrate, and filtrate is used successively
Watery hydrochloric acid, saturated sodium bicarbonate solution, saturated common salt water washing, are dried precipitation, obtain final product dichloroacetyl product.
Alternative implementation as the present embodiment, reaction temperature could alternatively be the arbitrary value in the range of 0-45 DEG C,
Have no effect on the realization of the object of the invention.
The dichloroacetylization of embodiment 2 is reacted
The present embodiment is the dichloroacetylization reaction in the method for prepare the chloramphenicol, specific as follows:
In the 250mL four-necked bottles equipped with agitator, thermometer and dropping funel, 21.6g is put into nitro-alpha-amido benzene
Acetophenone hydrochloride and dichloromethane 150mL, are cooled to less than 5 DEG C after stirring, then add dichloroacetyl chloride under agitation
14.5g, separately takes triethylamine solution 21g, is slowly dropped at a temperature of 15 DEG C in reactant liquor with dropping funel, is then warmed up to 15
DEG C, 1h is stirred, after completion of the reaction, filter, then filter cake is washed with dichloromethane, filtrate is collected, and filtrate is used successively dilute salt
Acid, saturated sodium bicarbonate solution, saturated common salt water washing, are dried precipitation, obtain final product dichloroacetyl product 28.2g, yield 97%.
The aldol reaction of embodiment 3
The present embodiment is the aldol reaction in the method for prepare the chloramphenicol, specific as follows:
The dichloroacetyl product prepared in 29.1g embodiments 2 is mixed with 200ml ethanol, 11.4g formaldehyde
Uniformly, the pH value for adjusting mixed solution is 7.2, is reacted at 35 DEG C of temperature, and reaction is finished, obtained final product to nitro-α-dichloroacetyl
Amido-beta-hydroxyphenyl acetone.
Be preferably carried out mode as the present embodiment, it is above-mentioned after completion of the reaction, reaction mixture is cooled to into 0 DEG C, mistake
Filter, is washed, suction filtration with frozen water, is dried, and is obtained final product to nitro-α-dichloroacetyl amido-beta-hydroxyphenyl acetone.
Used as the interchangeable embodiment of the present embodiment, the reaction temperature of the aldol reaction could alternatively be 32-
Arbitrary value in the range of 35 DEG C, pH value can be replaced the arbitrary value in the range of 7.2-7.5, have no effect on the object of the invention realization.
The aldol reaction of embodiment 4
The present embodiment is the aldol reaction in the method for prepare the chloramphenicol, specific as follows:
In equipped with agitator, the 250mL three-necked bottles of thermometer, described two prepared in 29.1g embodiments 2 are added
Chloroacetylation product and the formaldehyde 11.4g that ethanol 200mL, mass fraction are 37%, use a small amount of NaHCO after being well mixed3Saturation
It is 7.5 that solution adjusts the pH value of mixed solution, then, 32 DEG C is to slowly warm up under agitation, after completion of the reaction, rapid cold
But to 0 DEG C, filter, washed with frozen water, then suction filtration, be dried, obtain final product to nitro-α-dichloroacetyl amido-beta-hydroxyphenyl acetone
29.35g, yield 92%.
The catalytic reduction reaction of embodiment 5
The present embodiment is the catalytic reduction reaction in the method for prepare the chloramphenicol, specific as follows:
By the nitro-α prepared in 50g embodiments 4-dichloroacetyl amido-beta-hydroxyphenyl acetone and 1g ketone also
(buying is from Suzhou pilotage bio tech ltd for protoenzyme:Goods number YH2069, only provides the product of one of which model herein
Product are illustrated the effect of the present invention, between the product of commercially available each model for the purpose of the present invention is realized and indifference,
Similarly hereinafter, it is no longer redundant later hereinafter) be added to buffer solution, temperature be 40 DEG C, pH value be to react under conditions of 8.0, reaction is finished
Obtain final product the chloramphenicol.
As the present embodiment preferred embodiment, glucose, GDH have been additionally added in the buffer solution with
And NADP+, as the specific implementation of the preferred embodiment, the glucose, GDH and NADP+Use
Amount is respectively 62.5g, 1g and 50mg.
It should be noted that in the present embodiment, the buffer solution is PBS cushioning liquid, the PBS cushioning liquid can also be replaced
Other cushioning liquid are changed to, as long as can play and keep salt balance, the cushioning effect of the appropriate pH of adjustment in enzymic catalytic reaction
.
Alternative implementation as the present embodiment, the reaction temperature of the catalytic reduction reaction could alternatively be 20-
Arbitrary value in the range of 40 DEG C, the pH value of reaction solution could alternatively be the arbitrary value in the range of 6.0-8.0, have no effect on this
The realization of improving eyesight, similarly hereinafter, repeats no more hereinafter.
The catalytic reduction reaction of embodiment 6
The present embodiment is the catalytic reduction reaction in the method for prepare the chloramphenicol, specific as follows:To make in embodiment 4
For three mouthfuls of burnings that the nitro-α-dichloroacetyl amido for obtaining-beta-hydroxyphenyl acetone 50g, glucose 62.5g is placed in 1000mL
Bottle, then be added thereto to 500mL pH value be 6.5 and concentration for 0.05mol/L PBS, then, there-necked flask is put
In entering reaction pot, rotating speed 850rpm, 30 DEG C of temperature, then the NADP for being separately added into 50mg thereto are set+, the glucose dehydrogenation of 1g
(buying is from Suzhou pilotage bio tech ltd for enzyme:Goods number YH1901, only provides the product of one of which model herein
Illustrated the present invention effect, between the product of commercially available each model for the purpose of the present invention is realized and indifference), with
And 1g ketoreductase powder (is purchased from Suzhou pilotage bio tech ltd:Goods number YH2069), mixed solution is obtained, adopt
Make the pH value of the mixed solution maintain 6.5 with the NaOH solution that concentration is 2mol/L, react 3h, obtain final product the chloramphenicol,
And measure reaction conversion ratio>99%,>99%Ee,>99:1Dr, (wherein Ee=(R, R)/[(R, R)+(S, S)];Dr=[(R, R)
+(S,S)]/[(R,S)+(S,R)])。
As the preferred embodiment of the present embodiment, reacted using HPLC monitoring, extend reaction time conversion ratio and no longer increase
Plus, that is, judge that reaction is finished.
It is anti-with dichloromethane extraction after the catalytic reduction reaction is finished as the preferred embodiment of the present embodiment
Should after solution, merge organic phase, and be dried, precipitation, that is, obtain the chloramphenicol.Above-mentioned preferred reality is adopted in the present embodiment
Mode is applied, 47 grams of the chloramphenicol, yield 94%, purity 98%, Ee is obtained>99%, Dr>99:1.
It should be noted that the pH value of the PBS cushioning liquid can be replaced the arbitrary value in 6.0-7.5, concentration can be replaced
The arbitrary value being changed in 0.05-0.1mol/L, has no effect on the realization of the purpose of the present invention.
Obviously, above-described embodiment is only intended to clearly illustrate example, rather than the restriction to embodiment.For
For those of ordinary skill in the art, the change or change of other multi-forms can also be made on the basis of the above description
It is dynamic.There is no need to be exhaustive to all of embodiment, and the obvious change thus extended out or change
Among moving still in the protection domain of the invention.