KR100400885B1 - Separating method of single isomer from the ketorolac of the racemic body - Google Patents

Abstract

The present invention relates to a method for separating (-)-isomer ketorolac, which is known to be pharmacologically effective in the human body in ketorolac (Cetorolac) present in the form of racemates, In particular, it is an object to provide a novel separation method capable of separating single isomers of racemate ketorolac with high purity.

According to one aspect of the present invention, a ketorolac is reacted with an alcoholic compound in the presence of a specific enzyme catalyst, and then the ketorolac is separated from the (-)-isomer in the reaction mixture, or A novel separation method is disclosed in which a (-)-isomer ketorolac is separated from a mixture obtained by reacting a ketorolac ester compound obtained by reacting with an alcohol compound in the presence of a specific enzyme catalyst.

Description

Separating method of single isomer from the ketorolac of the racemic body}

The present invention relates to a method of purely separating ketoorolac of racemates into a single isomer, and in particular, using an enzyme such as Gindada Entatica lipasebi as a catalyst. It is about how to isolate.

Recently, many techniques using an enzyme as a catalyst for the preparation of specific compounds in the field of fine chemistry are used due to the reaction specificity of the enzyme and environmentally friendly reaction conditions of the manufacturing process. Among enzyme catalysts, lipases have been used in various ways because they have the advantages of low cost due to the simple process of producing enzymes without the help of coenzymes. Among them, lipase is widely used in the production of lipids in the food industry, etc., and is known to be used for producing about 60 million tons of lipids per year.

In addition, the lipase has a characteristic of selectively reacting with a compound containing an asymmetric carbon to selectively obtain only an active ingredient, which is a reaction to partially or totally hydrolyze lipids into fat and glycerol, intermolecular esterification

It is mainly applied to an interesterification reaction, a transesterification reaction, or a synthesis reaction of an ester compound.

Ketorolac (5-benzoyl-1,2-dihydro-3H-pyrrolo [1,2-a] pyrrole-1-carboxylic acid), which is the target compound of the present invention, has a molecular formula of the following general formula (1) It is a compound containing one asymmetric carbon.

Ketorolac having the structure of Chemical Formula 1 is present in the form of a mixed racemate of (+)-isomer and (-)-isomer which are optical isomers, among which (-)-isomer is pharmacologically human (+)-Isomers are known to have no pharmacological efficacy at any dose, and the biological differences between these isomers are platelet aggregation inhibition, prostaglandin synthesis, and anti-inflammatory analgesic analysis. It has been confirmed by various pharmacological efficacy tests such as analgesic and antiinflammatory analysis.

Ketorolac is particularly used as a drug such as nonsteroidal anti-inflammatory drugs. Recently, various disorders, including digestive disorders, hematologic disorders, hypersensitivity disorders, and kidney disorders, have been identified in patients using these drugs. Prescriptions such as minimizing the dosage and shortening the taking time are used as a way to overcome the disorder.

One method of minimizing the dose of ketorolac is to use a single isomer of the (-)-isomer to achieve a 50% reduction compared to the previous dose, and to have no pharmacologically identified (+)- The advantage is that the isomers can be removed beforehand.

Little is known about the separation of single isomers of ketorolac to date, only by Gerd Fulling and Charles J.Sih in 1987 by J. Am. Chem. Soc. (1987, 109, 2845) is the only one published. It is presumed that the lack of studies on the separation of isomers of ketorolac is due to the high tendency of racemization of the compounds themselves.

In this paper, Huling and Shi have published the results of a method for isomer separation using various hydrolytic enzymes using ketorolac ester as a substrate compound. Here, the hydrolytic enzymes include lipase-based enzymes such as Mucomihai, Pseudomonas, Candida Lugosa, and Posinpancreases, as well as Streptomyces gryeus, Aspezirus cytos, Bactus certilis, and Aspergillus oriza. The separation process was performed using protease-based enzymes as catalysts. In this case, when the protease-based enzymes were used as catalysts, excellent results were obtained in which the purity of the (-)-isomer was 95% or more. When enzymes were used as catalysts, it was difficult to obtain the (-)-isomer in high purity.

It is an object of the present invention to provide Candida antartica lipase B (Candida antartica lipase B), which is obtained from Candida entatica, among lipase family enzymes known to have little effect as an enzyme catalyst for the separation of such single isomers. It is optionally used as an enzymatic catalyst for the separation of single isomers to separate high purity negative (-)-isomers.

It is another object of the present invention to separate ketorolac, a single isomer of (-)-isomer, with high purity by using an ester derivative of ketorolac as well as an acid compound ketorolac as a substrate compound.

The present invention relates to the reaction of a ketorolac ester compound with a (-)-isomer of ketorolac as shown in Chemical Formula 2 by reacting ketorolac with an alcohol-like compound under a specific condition. It is characterized in that the ketorolac of the (-)-isomer is separated through a separation process after preparing the mixed product.

The present invention is also carried out by subjecting the racemic ketorolac ester compound obtained by reacting the racemate ketorolac and an alcohol compound to the substrate compound and performing a hydrolysis reaction using Candida entica lipase as an enzyme catalyst. Another characteristic is to separate the (-)-isomer ketorolac by preparing a mixed product of the (-)-isomer and the ketorolac ester compound, as shown in Chemical Formula 3 below. It is.

[Chemical Scheme 2]

R is an alkyl group having 3 to 20 carbon atoms or an aryl group

[Chemical Scheme 3]

R is an alkyl group having 3 to 20 carbon atoms or an aryl group

Hereinafter, the present invention will be described in detail.

Candida entatica rivazebi used as a catalyst in the present invention is an enzyme catalyst obtained from a lipase-based Candida entatica using NovoDisk, etc., and the preferred amount in the present invention is ketorolac as the substrate compound. It is in the range of approximately 10 to 150% by weight relative to the amount of lock used, and there is a problem in that enantioselectivity is lowered when using less than 10% by mass, and when using more than 150% by mass, there is almost no catalyst improvement effect and only a cost increase. In addition, when using a ketorolac ester compound as a substrate compound, it is preferable to use a ribase ratio in the range of about 10-100 mass% with respect to the total usage-amount of a ketorolac ester compound.

In the present invention, when ketorolac is used as a substrate compound, the ketorolac is reacted with an alcohol compound and a Candida entatica lipase catalyst and a ketorolac ester compound and a mixture of (-)-isomer form ketorolac in specific conditions. Alcohol compounds used at this time are, for example, ethanol, n-butanol, n-hexanol, n-octanol, n-decanol, n-tetra decanol, 2-chloroethanol, benzyl alcohol, iso Alcohol compounds having 3 to 20 carbon atoms are preferable, such as propyl alcohol and isobutyl alcohol, and more preferably alcohol compounds having 4 to 10 carbon atoms, and particularly preferably n-octanol having 8 carbon atoms.

Specific reaction solvents include acetonitrile, diisopropyl ether, dichloromethane, 1,2-dichloroethane, tetrahydrofuran, dimethylformamide, diisopropyl ether, dichloromethane and 1,2-dichloroethane. It showed better performance, especially 1,2-dichloroethane is recommended because it has the advantage of reducing the reaction time.

The temperature suitable for the reaction is in the range of approximately 20 to 70 ° C, more preferably in the range of 30 to 60 ° C.

On the other hand, the present invention by using a ketorolac ester compound as a substrate compound and a lipase ratio as an enzyme catalyst by a hydrolysis reaction of the (-)-isomer ketorolac and unreacted ketorolac as shown in the chemical formula 3 Another feature is to prepare a mixture of ester compounds to separate the materials to obtain a (-)-isomer ketorolac. The reaction solution used in the hydrolysis reaction is approximately pH 6-9 (more preferably pH 7). It is preferable to use a buffer in the range, and specific examples of the buffer solution include phosphate buffer and the like. In addition, a preferable reaction temperature in the hydrolysis reaction is in the range of about 10 to 40 ° C (more preferably, 20 to 30 ° C).

The acid liberated during the hydrolysis reaction is preferably neutralized with a dilute alkali solution.

As described above, a method of preparing a mixture of a ketorolac ester compound and a (-)-isomer ketorolac using a ketorolac and a ketorolac ester compound as a substrate compound of the reaction, and then separating them is specified in the present invention. Although not preferred, adsorptive separation using HPLC or LC is preferred.

Hereinafter, the present invention will be described in more detail with reference to Examples.

Example 1

<1> Ketorolac Butyl Ester Synthesis

Ketorolac 100mg (0.39m mol) was added to a mixture of 2 mL butanol and 0.1 mL concentrated sulfuric acid and reacted under reflux for 3 hours. The reaction solution was cooled to room temperature, diluted with ice water, and extracted with diethyl ether. After neutralizing the organic layer with an aqueous solution of sodium hydrogen carbonate, the organic layer was separated, dehydrated, filtered and concentrated, and then quantitatively obtained ketorolac butyl ester by using liquid chromatography (n-hexane: ethyl acetate = 10: 1).

<2> Hydrolysis of Ketorolac Butyl Ester Using Lipase

40 mg (0.16 mmol) of ketorolac butyl ester obtained in the above <1> was added to 7 ml of buffer solution (pH7), and 20 mg of Candida Entica Lipase ratio was added thereto, and the reaction was performed at a temperature of approximately 25 ° C. At this time, the acid liberated during the reaction was automatically neutralized with 0.02N caustic soda solution, and samples were collected at regular time intervals from the reaction solution, and high performance liquid chromatography (HPLC, mobile phase: n-hexane / isopropanol / trifluoroacetic acid) was used. = 90/10 / 0.1) and stationary phases (chiralpak AD) were used to measure acid enantioselectivity and the results are shown in Table 1 below with the total reaction time and the absolute structure of the obtained ketorolac.

Comparative Example 1-5

Except that shown in Table 1 as an enzyme catalyst in Example 1 was used in the same manner as in Example 1 except that the same molar amount was used, and the results are shown in Table 1.

TABLE 1

division Enzyme catalyst Response time (hr) Acid Mirror Image Selectivity (%) Absolute structure Example 1 Comparative Example 1 Comparative Example 2 Comparative Example 3 Comparative Example 4 Comparative Example 5 Candida Entica Lipabie Candida Lugosa Lipaje Coltria Sephacia Streptomyces Griseus Aspergillus Cytomumuko Mihai 0.56.857.31.3522.30.5 10091831006471 RRRSRR

R is the (+)-isomer and S is the (-)-isomer

Example 2

10 mg (0.039 mmol) of ketorolac and 0.039 mmol of butanol were dissolved in 5 ml of acetonitrile and 30 mg of molecular sieve (4 cc) was added thereto.

10 mg of Candida Entica Lipasebi was added thereto, followed by a holding reaction at 34 ° C. Samples were taken from the reaction solution at regular time intervals to obtain high-performance liquid chromatographs (HPLC, mobile phase: n-hexane / isopropanol / trifluoroacetic acid = 90/10 / 0.1) and stationary phase (chiralpak AD) and reaction mirror images. The physical properties such as selectivity are measured and shown in Table 2 below, and in the same process except that other alcohol compounds shown in Table 2 are used in equimolar amounts instead of butanol, and the results are shown in Table 2. It was.

TABLE 2

Alcohol compounds Response time (h) % Conversion Ester Mirror Image Selectivity (%) Absolute structure Ethanolbutanolhexanoloctanoldecanoltetradecanol2-chloroethanolphenylmethanolisopropanolisobutanol 434248525249254811649 435359585958213056 52908693878647802576 SSSSSSSSSS

Example 3

10 mg (0.039 mmol) of ketorolac and 0.39 mmol of octanol were dissolved in 5 ml of acetonitrile and 30 mg of molecular sieve (4 cc) was added thereto. 10 mg of Candida Entica lipase ratio was added to the reaction solvent, and the reaction was carried out while stirring at 34 ° C. Samples were taken from the reaction solution at regular intervals, and then converted into high-performance liquid chromatography (HPLC, mobile phase: n-hexane / isopropanol / trifluoroacetic acid = 90/10 / 0.1) and stationary phase (chiralpak AD) and ester mirror images. The results of the measurement of selectivity and the like are shown in Table 3 below, and the same results as in Table 3 were used except that the solvents shown in Table 3 were used instead of the acetonitrile used as the reaction solvent. Indicated.

TABLE 3

Solvent Response time (h) % Conversion Ester Mirror Image Selectivity (%) Absolute structure Acetonitrile diisopropyl ether dichloromethane tetrahydrofuran n-hexanedimethylformamide 1,2-dichloroethane 52204869696969 5852541488051 9388100131006100 SSSSSSS

As can be seen from the above examples and comparative examples, in the case of separating keratolac of racemate into a single isomer according to the present invention, it is possible to separate not only high-purity (-)-isomer compared to the conventional method, but also to separate Usability can be obtained, such as to shorten the reaction time required for the process.

Claims (8)

A keroserol derivative of the racemate characterized by separating the (-)-isomer ketorolac from the mixture obtained by reacting the ketorolac of the racemate represented by the following formula (1) in the presence of an alcohol compound and Candida Entica lipase ratio. How to separate Torolac into a single isomer. [Formula 1] The (-)-isomer ketorolac is separated from the mixture obtained by hydrolyzing the racemate ketorolac ester with the alcoholic compound in the presence of Candida entica lipase ratio. A method for separating ketorolac of racemates into a single isomer, characterized in that it is carried out. The method according to any one of claims 1 and 2, wherein the alcohol compound is an alcohol compound having 3 to 20 carbon atoms. The method of claim 1, wherein the amount of candida enteric lipase is in the range of 10 to 150% by weight based on the total amount of ketorolac. The method for separating ketorolac of racemates into single isomers according to claim 2, wherein the amount of candida enteric lipase is in the range of 10 to 100% by mass based on the total amount of the ketorolac ester compound. . The method according to claim 1, wherein the reaction temperature of the ketorolac and the alcohol compound is in the range of 20 to 70 ° C. 3. The method of claim 2, wherein the buffer used for hydrolysis is a buffer in the range of pH 6-9, wherein the ketorolac of the racemate is separated into a single isomer. 3. The racemate according to any one of claims 1 and 2, characterized in that an adsorptive separation method is used to separate the (-)-isomer of the ketorolac in a mixture of the ketorolac and the ketorolac ester compound. Method for separating ketorolac into single isomers.