KR20090083742A - Chiral stationary phase based on antibiotics cefaclor and chiral column - Google Patents

Abstract

A chiral stationary phase of LC (liquid chromatography) using cefaclor derivative which is an antibiotics suppressing biosynthesis of cell wall is provided to isolate a racemic compound containing a pharmaceutical product having optical activity. A chiral solid phase (CSP1) based on a cepaclo which is cephalosporin based antibiotics is denoted by the chemical formula 1. In the chemical formula 1, R is hydrogen, one or two methyl group, one or two methoxy group or t-butyl group, nitro group, and chloro group. The chiral solid phase(CSP1) ia a novel pirkle chiral solid phase which is applicable to the LC. A method for manufacturing the chiral solid phase of the chemical formula 1 comprises: a step of condensing a compound of the chemical formula 3 with pharmaceutical product having optical activity of the chemical formula 2 to obtain a compound 4; a step of using EEDQ (2-ethoxy-1-ethoxycarbonyl-1,2-dihydroquinoline); and a step of reacting the compound 4 with a silica gel to obtain the chiral solid phase of the chemical formula 1. The chiral column is filled with the chiral solid phase of the chemical formula 1.

Description

Chiral stationary phase based on antibiotics cefaclor and chiral column

The present invention relates to a chiral stationary phase for LC and an optical separation of chiral pharmaceuticals and a chiral column filled with them, and more specifically, to separate two optical isomers constituting racemic bioactive substances (including racemic pharmaceuticals). It relates to a Pirkle-type chiral stationary phase which is known to be very useful and a chiral column packed with it.

Many drugs are currently used in racemic mixtures, but the two enantiomers of this racemic combination can be broken down, absorbed and activated at different rates after administration.

In addition, these two enantiomers may have the same medicinal function, but one component may act as a useful drug while the other may not act as a drug or in some cases may be harmful.

For example, in the case of ibuprofen, the S-(+)-form is anti-inflammatory, but in the R-(+)-form, it causes side effects.In the case of Pemicilamine, the D-form is effective for treating rheumatism, but the L-form is For this reason, it is urgent to produce a purely isolated single enantiomer for the purpose of improving the efficacy and stability of the drug.

The transition from racemic mixtures to single enantiomer drugs is exponentially growing in demand, with the policy of 'It is appropriate to produce a single enantiomer chiral drug' published by the FDA in 1992. The enantiomer's chiral drug development direction is being actively researched for the development of new synthetic methods and for the separation of existing racemic mixtures.

The pair of optical isomers in the mirror image of the racemic compound have the same physical and chemical properties due to the difference in the spatial arrangement of the atoms or functional groups in the molecular structure of the compound, and thus cannot be distinguished by a general method. It can only be distinguished under a chiral environment.

Since the living body is composed of (L) -amino acids and (D) -carbohydrates having optical activity, they are essentially chiral and act as a chiral environment. Therefore, it is known that a pair of optical isomers (racemic bioactive materials) are distinguished in vivo and exhibit different biological activities.

In particular, one of the two optical isomers that constitute the racemic drug exhibits the desired pharmacological action in the human body, but the other optical isomer, which is a mirror image thereof, may have side effects such as toxicity. In the process of authorizing a new drug (including pesticides or food additives), the FDA does not permit the manufacture and sale of a racemic product if the drug does not provide sufficient basis for use in the racemic form. Regulations are in place.

As a result of this regulation, pharmaceutical companies around the world are making great efforts and investments in research on the difference in clinical efficacy between racemic and pure optical isomeric drugs, and separate the two optical isomers that make up the racemic drugs. BACKGROUND ART There is an increasing demand in the fields of chemistry, medicine, and pharmacy for a technique for rapidly and accurately measuring optical purity of optically active pharmaceuticals composed of branched optical isomers.

The chiral stationary phase based on the optically active N, N'-di (substituted benzoyl) -2,3-diamino-1,4-butadiol of No. 0525288 registered by the inventor of the present invention and filled with these In chiral columns, the pi-pi interaction plays an important role and is related to the chiral stationary phase that optically splits racemic compounds.

Through the construction according to the prior art, the conventional invention is C ₂ symmetric and optically active (2R, 3R) -2,3-diamino-1,4-butadiol or (2S, 3S) -2,3 -Diamino-1,4-butadiol is made of a bezoyl derivative in which an aromatic ring is appropriately substituted with two amino groups, and the chiral stationary phases covalently bonded to silica gel for LC and the chiral column for LC filled with them are provided. The present invention provides a method for optically separating racemic compounds having aromatic groups in a molecule by stereoselective pi-pi interaction between chiral stationary phases and racemic compounds to be optically divided.

This conventional invention is an optical splitting of racemic bioactive material using a chiral column for LC filled with a chiral stationary phase for LC among various techniques that can accurately measure the optical purity of a chiral bioactive material including an optically active chiral drug. The technology not only provides a technique for acquiring two optical isomers, but also provides a technique for measuring optical purity of an optically active bioactive substance, but a technique for measuring optical purity more accurately is available. There is a problem.

Pirkle-type chiral stationary phases that have been developed and used in the past have been prepared by immobilizing 3,5-dinitrobenzoyl derivatives of mainly optically active amino acids on silica gel.However, the racemate mixture is highly separated and redeveloped as a single enantiomer drug. More effective chiral separation techniques are needed in racemic switches.

An object of the present invention is to provide a novel Pirkle-type chiral stationary phase, which is based on a derivative of Sephacller, a Cephalosporin-based antibiotic belonging to the beta lactam system of the present invention.

It is another object of the present invention to provide an LC column comprising the new chiral stationary phase described above.

Another object of the present invention is to provide a method for separating two optical isomers constituting a racemic bioactive material including a racemic drug using an LC column including a new stationary phase.

In order to achieve the above object, the present invention provides a chiral stationary phase (CSP1) represented by the following chemical formula:

Where

R is characterized by being hydrogen, one or two methyl groups, one or two methoxy groups, one or two t-butyl groups, one or two nitro groups, one or two chloro groups.

Here, the present invention is characterized in that the chiral column filled with a chiral stationary phase (CSP 1) represented by the formula (1).

The present invention has the effect of providing a novel optical splitting method for effectively separating the racemic compound including the drug having optical activity by using a chiral column for HPLC filled with a chiral stationary phase (CSP 1).

In addition, the present invention provides an analytical technique necessary for the optical purity measurement technology of chiral drugs because the optical splitting of the 1,4-benzodiazepin-2-one derivatives used as anti-anxiety agents or neurostabilizers can be more effective than the baseline separation. It is effective.

Sephacler benzoyl derivative fixed phase according to the present invention is capable of stereoselective π-π interaction with various amino acid derivatives including 1,4-benzodiazepine derivatives and benzoyl groups, and can be separated separately by HPLC separation using the same. have. Sephacler benzoyl derivative stationary phases can be used as π-base selectors and π-acid selectors depending on the π-electron density of the benzoyl group, and this property is characterized by the It is believed that the difference results in retention time differences that can be selectively separated.

Therefore, in the following formula (CSP 1), the functional group which affects the π-cell density of the benzoyl group of the sepachlor benzoyl derivative stationary phase according to the present invention, R is hydrogen, one or two methyl groups, one or two methoxy groups, one or It may be limited to two t-butyl groups, one or two nitro groups, one or two chloro groups.

The present invention provides a benzoyl derivative in which an aromatic ring is appropriately substituted with an amino group of a chiral stationary phase (CSP 1), an optically active drug, Sephacle, represented by the following formula (1), and then covalently bonded to the silica gel for LC to form the chiral stationary phase ( It provides a method for producing CSP 1), a chiral column filled with the chiral stationary phase (CSP 1), and an optical splitting method of a racemic bioactive material using the same.

Wherein R represents hydrogen, one or two methyl groups, one or two methoxy groups, one or two t-butyl groups, one or two nitro groups, one or two chloro groups.

Referring to the present invention in more detail as follows.

In Scheme 1, R is as defined above.

The compound represented by Chemical Formula 1 according to the present invention may be prepared by the same method as in Scheme 1. Compound 4 is obtained by condensing the compound represented by Chemical Formula 3 using the optically active drug Sephacleler represented by Chemical Formula 2 as a starting material in Scheme 1. To this, Compound 4 obtained by introducing 3-aminopropyltriethoxysilane in a dichloromethane solvent using EEDQ (2-ethoxy-1-ethoxycarbonyl-1,2-dihydroquinoline) as a coupling reagent is obtained. . This is reacted with silica gel to obtain a chiral stationary phase represented by formula (1).

The solvent used to prepare the stationary phase is not particularly limited as long as it can effectively perform the constant boiling effect (azeotrope effect) to remove the moisture of the silica gel, for example, benzene, toluene may be used.

On the other hand, in the above, the coupling agent is generally an eye coupling agent, for example EEDQ (2-ethoxy-1-ethoxycarbonyl-1,2-dihydroquinoline), EDC (1-ethyl-3- (3) -Dimethylaminopropyl) -carbodiimide), ethylchloroformate and the like can be used.

Meanwhile, according to the present invention, the compound 4 has a physical property of having viscosity due to covalent bonding of the compound 4 with silica gel without undergoing a pretreatment process. After the reaction is completed, it is preferable to use a filtering method using 1N hydrochloric acid aqueous solution to increase the physical properties of the silica gel.

On the other hand, the surface concentration of the finally prepared Sephacler benzoyl derivative fixed phase represents the amount of reactants combined with the silica gel, and the surface area of the silica gel varies depending on the type of silica gel produced, but in general, the larger the surface area, the greater the number of reactants. .

In addition, the Sephacler benzoyl derivative fixed phase according to the present invention has a stereoselective π-π interaction, which contributes to the optical separation of the analyte and the fixed phase. As an eluent, a nonpolar solvent such as n-hexane, n-pentane, n Preference is given to using -heptanes and polar solvents such as ethanol, 2-propanol in appropriate proportions.

The chiral stationary phase represented by Chemical Formula 1 (CSP1) prepared by the above manufacturing method is suspended in methanol, and filled in a hollow column for LC using a slurry filler to prepare a chiral column.

The present invention as described above will be described in more detail the present invention through the following examples, but the following examples are for illustrative purposes of the invention and are not limited by the appended claims.

Example  1.N- (3 ', 5'- Dimethoxybenzoyl )-Sephacler (Formula 3; R = OCH ₃ Manufacturing

Dissolve Cefaclor (2g, 5.44mmol) in purified 25ml THF in a 100ml round bottom flask, cool to 0 ° C, stir, slowly add Propylene oxide (1.14ml, 16.31mmol) with a syringe, then stir for 30 minutes and THF 20ml 3,5-Dimethoxybenzoyl chloride (1.20g, 5.98mmol) dissolved in was slowly dropped through a syringe and allowed to react for 20 hours at room temperature under argon.

After completion of the reaction, the solvent was removed with a reduced pressure evaporator and recrystallized with THF and Ethyl acetate to give yellowish white crystals. 2.66 g (yield 89%) of white solid compounds were obtained.

¹ H-NMR (Acetone-d6, ppm): δ = 3.57 (d, J = 18.6 Hz, 1H), 3.81 (t, J = 15 Hz, 6H), 3.93 (d, J = 18.6 Hz, 1H), 5.22 (d, J = 5.1 Hz, 1H), 5.91 (m, 1H), 5.95 (d, J = 2.4 Hz, 1H), 6.63 (qt, J = 3.9 Hz, 1H) 7.10 (qd, J = 2.4 Hz, 2H), 7.34 (m, 2H), 7.38 (m, 2H), 7.58 (m, 1H), 8.42 (s, 3H); FT-IR (KBr): ～ v = 3280, 3120, 1790, 1590, 1160, 714 cm ^-1 mp = 159.7 ~ 163.9 ° C [α] _D ^{14 .5} 51.29 (c = 0.11, CHCl ₃ )

Example  2.N- (3 ', 5'- Dimethoxybenzoyl )- N ' - Aminopropyltriethoxysilane Sephacleler (Formula 4; R = OCH ₃ )of  Produce

 In a 100 ml round bottom flask, compound (0.50 g, 1.00 mmol) and EEDQ (0.27 g, 1.10 mmol) obtained in the same reaction as in Example 1 were added, and 40 ml of THF purified by needle was added while stirring under argon. give. AMPS (3-Aminopropyl triethoxysilane) (0.26ml, 1.10mmol) was added thereto using a syringe and reacted for 6 hours at room temperature under argon. The resulting compound proceeds to the next reaction without workup.

¹ H-NMR (Acetone-d6, ppm): δ = 1.93 (m, 2H), 2.89 (m, 4H), 3.52 (d, J = 8.1Hz, 1H), 3.81 (t, J = 26.1Hz, 6H ), 3.81 (d, J = 26.2 Hz, 1H), 5.22 (d, J = 23.7 Hz, 1H), 5.80 (m, 1H), 5.91 (d, J = 4.8 Hz, 1H), 6.63 (qt, J = 1.8 Hz, 1H) 7.10 (qd, J = 17.7 Hz, 2H), 7.38 (m, 2H), 7.53 (m, 2H), 7.60 (m, 1H), 8.35 (s, 3H) FT-IR (KBr ): ～ v = 3280, 2970, 1600, 1070, 766 cm ^-1 mp = 172.4 ~ 175.1 ℃ [α] _D ^{14 .5} 51.29 (c = 0.11, CHCl ₃ )

Example  3. Chiral  (Formula 1; R = OCH ₃ Manufacturing

Place a Dean-stack trap and reflux in a 500 ml two-necked round bottom flask. 2.8 g of silica gel (Kromasil 5 μm; 100) for HPLC column was added thereto, and 130 ml of toluene was added thereto, and the mixture was refluxed under argon to remove H ₂ O adsorbed on the silica gel using a Dean-stack trap. The compound is added by needle and refluxed for 72 hours. After the reaction, the vessel was cooled sufficiently, and the reaction mixture was filtered under reduced pressure to recover silica gel. After sufficient drying in a vacuum pump and oven, the CSP was sonicated with methanol and packed into a 4.6 mm × 150 mm stainless empty column for HPLC. According to trace element analysis (Found: C, 13.66: H, 1.82: N, 3.68), based on carbon content, 0.45 mmol of chiral selector per g of silica gel or 2.06 μm of chiral selector per m 2 of silica surface It was confirmed to be fixed.

Example  4.N- (3 ', 5'- Dinitrobenzoyl )-Sephacler (Formula 3; R = NO ₂ Manufacturing

Dissolve Cefaclor (2g, 5.44mmol) in purified 25ml THF in a 100ml round bottom flask, cool to 0 ° C, stir, slowly add Propylene oxide (1.14ml, 16.31mmol) with a syringe, then stir for 30 minutes and THF 20ml 3,5-Dinitrobenzoyl chloride (1.38g, 5.98mmol) dissolved in was slowly dropped through a syringe and allowed to react for 20 hours at room temperature under argon.

After completion of the reaction, the solvent was removed with a reduced pressure evaporator and recrystallized with THF and Ethyl acetate to give yellowish white crystals. 1.61 g (yield 53%) of white solid compounds were obtained.

¹ H-NMR (Acetone-d6, ppm): δ = 3.55 (d, J = 18.6 Hz, 1H), 3.94 (d, J = 18 Hz, 1H), 5.23 (d, J = 5.1 Hz, 1H), 5.92 ~ 5.96 (m, 1H), 5.98 (d, J = 4.8 Hz, 1H), 7.36-7.39 (m, 2H), 7.39-7.43 (m, 1H), 7.59-7.82 (m, 2H), 8.54 (s , 3H), 9.08 (dd, J = 2.1 Hz, 2H), 9.16 (q, J = 1.8 Hz, 1H); FT-IR (KBr): ～ v = 3270, 3150, 1790, 1540, 1350, 727 cm ^-1 mp = 212.1 ~ 216.7 ° C [α] _D ^{14 .5} 42.46 (c = 0.04, CHCl ₃ )

Example  5.N- (3 ', 5'- Dinitrobenzoyl )-Sephacler (Formula 4; R = NO ₂ Manufacturing

In a 100 ml round bottom flask, compound (0.50 g, 1.00 mmol) and EEDQ (0.27 g, 1.10 mmol) obtained in the same reaction as in Example 4 were added, and 40 ml of THF purified by needle was stirred for 20 minutes while stirring under an argon stream. give. AMPS (3-Aminopropyl triethoxysilane) (0.26ml, 1.10mmol) was added thereto using a syringe and reacted for 6 hours at room temperature under argon. The resulting compound proceeded directly to the next reaction without workup.

¹ H-NMR (Acetone-d6, ppm): δ = 1.17 (m, 2H), 2.08 (m, 4H), 3.08 (d, J = 2.7 Hz, 1H), 3.58 (d, J = 18.6 Hz, 1H ), 5.23 (d, J = 4.2 Hz, 1H), 5.71-5.81 (m, 1H), 5.96 (d, J = 3.9 Hz, 1H), 7.37-7.43 (m, 2H), 7.49-7.53 (m, 1H), 7.58 ~ 7.63 (m, 2H), 8.32 (s, 3H), 8.91 (dd, J = 1.8Hz, 2H), 9.15 (q, J = 16.8Hz, 1H) FT-IR (KBr): ~ v = 3290, 2880, 1780, 1540, 787 cm ^-1 mp = 182.7-184.0 ° C. [α] _D ^{14 .5} 47.46 (c = 0.09, CHCl ₃ )

Example  6: Chiral  (Formula 1; R = NO ₂ Manufacturing

Place a Dean-stack trap and reflux in a 500 ml two-necked round bottom flask. 2.8 g of silica gel (Kromasil 5 μm; 100) for HPLC column was added thereto, and 130 ml of toluene was added thereto, and the mixture was refluxed under argon to remove H ₂ O adsorbed onto the silica gel using a Dean-stack trap. The compound is added by needle and refluxed for 72 hours. After the reaction, the vessel was cooled sufficiently, and the reaction mixture was filtered under reduced pressure to recover silica gel. After drying sufficiently in a pump and oven, this CSP was sonicated with methanol and packed into a 4.6 mm x 150 mm size stainless empty column for HPLC. According to trace element analysis results (Found: C, 4.01: H, 0.40: N, 0.48), based on carbon content, 0.12 mmol chiral selector per gram of silica gel or 0.44 μmol chiral selector per ㎡ surface of silica It was confirmed to be fixed at.

Example  7.N- (3 ', 5'- Dimethylbenzoyl )-Sephacler (Formula 3; R = CH ₃ Manufacturing

Dissolve Cefaclor (2g, 5.44mmol) in purified 25ml THF in a 100ml round bottom flask, cool to 0 ° C, stir, slowly add Propylene oxide (1.14ml, 16.31mmol) with a syringe, then stir for 30 minutes and THF 20ml 3,5-Dimethylbenzoyl chloride (1.01 g, 5.98 mmol) dissolved in was slowly dropped through a syringe and allowed to react for 20 hours at room temperature under argon.

After completion of the reaction, the solvent was removed with a reduced pressure evaporator and recrystallized with THF and Ethyl acetate to give yellowish white crystals. 2.26g (yield 83%) of yellowish white solid compounds were obtained.

¹ H-NMR (Acetone-d6, ppm): δ = 2.32 (q, J = 5.1 Hz, 6H), 3.53 (d, J = 5.1 Hz, 1H), 3.59 (d, J = 5.1 Hz, 1H), 5.21 (d, J = 4.8Hz, 1H), 5.91 (m, 1H), 5.93 (d, J = 2.7Hz, 1H), 7.17 (m, 1H), 7.34 (m, 2H), 7.39 (m, 2H ), 7.56 (m, 2H), 7.59 (m, 1H), 8.43 (m, 3H) FT-IR (KBr): ～ v = 3270, 1780, 1630, 698 cm ^-1 mp = 154-159.3 ° C [α ] _D ^{14 .6} 21.87 (c = 0.10, CHCl ₃ )

Example  8.N- (3 ', 5'- Dimethylbenzoyl )- N ' - Aminopropyltriethoxysilane Sephacleler (Formula 4; R = CH ₃ )of  Produce

In a 100 ml round bottom flask, compound (0.50 g, 1.00 mmol) and EEDQ (0.27 g, 1.10 mmol) obtained in the same reaction as in Example 4 were added, and 40 ml of THF purified by needle was stirred for 20 minutes while stirring under an argon stream. give. AMPS (3-Aminopropyl triethoxysilane) (0.26ml, 1.10mmol) was added thereto using a syringe and reacted for 6 hours at room temperature under argon. The resulting compound proceeded directly to the next reaction without workup.

¹ H-NMR (Acetone-d6, ppm): δ = 2.32 (q, J = 3.3Hz, 6H), 2.91 (m, 2H), 2.91 (m, 4H), 3.58 (d, J = 5.1Hz, 1H ), 3.59 (d, J = 5.1 Hz, 1H), 5.23 (d, J = 4.8 Hz, 1H), 5.96 (m, 1H), 5.99 (d, J = 2.7 Hz, 1H), 7.17 (m, 1H) ), 7.35 (m, 2H), 7.38 (m, 2H), 7.56 (m, 2H), 7.76 (m, 1H), 8.52 (m, 3H) FT-IR (KBr): ～ v = 3280, 1780, 1640, 696 cm ^-1 mp = 194.7 ~ 196.2 ° C [α] _D ^{14 .5} 49.23 (c = 0.11, CHCl ₃ )

Example  9: Chiral  (Formula 1; R = CH ₃ Manufacturing

Place a Dean-stack trap and reflux in a 500 ml two-necked round bottom flask. 2.8 g of silica gel (Kromasil 5 μm; 100) for HPLC column was added thereto, and 130 ml of toluene was added thereto, and the mixture was refluxed under argon to remove H ₂ O adsorbed onto the silica gel using a Dean-stack trap. The compound is added by needle and refluxed for 72 hours. After the reaction, the vessel was cooled sufficiently, and the reaction mixture was filtered under reduced pressure to recover silica gel. After sufficient drying in a pump and oven, the CSP was sonicated with methanol and filled into a 4.6 mm x 150 mm stainless empty column for HPLC. According to trace element analysis results (Found: C, 11.28: H, 1.64: N, 1.59), based on carbon content, 0.35 mmol chiral selector per gram of silica gel or 1.47 μmol chiral selector per 1 ㎡ silica surface was applied to silica gel. It was confirmed to be fixed.

Experimental Example  One : Chiral  (Formula 1: R = NO ₂ )this Filled LC for Chiral column  Α- Hydroxycarboxylidene  Optical Segmentation of Derivatives and 1,4-benzodiazepin-2-one Derivatives

 The developing solvent in the chiral column for LC using chiral stationary phase is 2-propanol / hexane = 20/80 or ethanol / hexane = 20/80, flow rate is 1 mL / min, and detector is 509 nm UV model 515 Α-hydroxycarboxanilide derivatives and 1,4-benzodiazepin-2-one derivatives were optically divided using HPLC equipment consisting of an HPLC pump, a Rheodyne model 7725i injector and a Waters 2487 absorbance detector, respectively, and are shown in Tables 1 and 2 below. It was.

TABLE 1 Chiral  ( CSP  1: R = NO ₂ )this Filled In chiral column  α- Hydroxycarboxylidene  Optical division of derivatives

R k ₁ k ₂ α CH ₃ 4.82 6.22 1.29 (CH ₃ ) ₂ CH 3.09 4.49 1.45 (CH ₃ ) ₂ CHCH ₂ 2.93 6.02 2.05 CH ₃ (CH ₂ ) ₃ 3.09 4.11 1.33 CH ₃ (CH ₂ ) ₅ 2.47 3.14 1.27 CH ₃ (CH ₂ ) ₇ 2.14 2.66 1.24 CH ₃ (CH ₂ ) ₉ 1.89 2.32 1.23 C ₆ H ₅ 5.27 7.76 1.47 C ₆ H ₅ CH ₂ 5.36 8.43 1.57

19.04 29.13 1.53

이동상 : 에탄올/헥산 = 20/80

Mobile phase: Ethanol / hexane = 20/80

k ₁ : capacity factor of the optical isomer initially eluted

k ₂ : capacity factor of the optical isomer eluting later

                  α: Separation factor

TABLE 2 Chiral  ( CSP  1: R = NO ₂ )this Filled In the key column  Optical Segmentation of 1,4-benzodiazepin-2-one Derivatives

R k ₁ k ₂ α CH ₃ 1.89 2.43 1.29 CH ₃ (CH ₂ ) ₂ 1.33 1.81 1.36 CH ₃ (CH ₂ ) ₃ 1.19 1.65 1.39 CH ₃ S (CH ₂ ) ₂ 2.48 3.41 1.37 (CH ₃ ) ₂ CH 1.13 1.61 1.42 (CH ₃ ) ₂ CHCH ₂ 1.23 1.78 1.44 C ₆ H ₅ CH ₂ 2.26 3.35 1.48

이동상 : 2-프로판올/헥산 = 20/80

Mobile phase: 2-propanol / hexane = 20/80

k _{1 ,} k _{2 ,} α are as defined in Table 1.

Experimental Example  2 : Chiral  (Formula 1: R = CH ₃ )this Filled LC for Chiral column  Used N- (3,5- Dinitrobenzoyl ) -Leucine Phenylalkylamide  Optical division of derivatives

 The developing solvent in the chiral column for LC using chiral stationary phase is 2-propanol / hexane = 20/80, the flow rate is 1 mL / min, and the detector is N- (3,5-dinitrobenzoyl) -Louis under conditions of 254 nm UV. Each of the phenylalkylamide derivatives was optically divided and shown in Table 3 below.

TABLE 3 Chiral  ( CSP  1: R = CH ₃ )this Filled In the key column  N- (3,5- Dinitrobenzoyl ) -Leucine Phenylalkylamide  Optical division of derivatives

R k ₁ k ₂ α CH ₃ 3.00 3.81 1.27 CH ₃ CH ₂ 2.78 2.93 1.06 CH ₃ (CH ₂ ) ₂ 2.21 2.38 1.08 CH ₃ (CH ₂ ) ₃ 1.81 2.16 1.19 CH ₃ (CH ₂ ) ₄ 1.62 1.98 1.22 CH ₃ (CH ₂ ) ₅ 1.49 1.84 1.23 CH ₃ (CH ₂ ) ₉ 1.11 1.56 1.41

k _{1 ,} k _{2 ,} α are as defined in Table 1.

Experimental Example  3: Chiral  (Formula 1: R = OCH ₃ )this Filled LC for Chiral column  Used N- (3,5- Dinitrobenzoyl ) -Leucine Phenylalkylamide  Optical division of derivatives

 N- (3,5-dinitrobenzoyl) -leucine phenyl in a chiral column for chiral column using ethanol / hexane = 20/80, flow rate 1 mL / min, detector at 254 nm UV The alkylamide derivatives were optically divided and shown in Table 3 below.

TABLE 3 Chiral  ( CSP  1: R = OCH ₃ )this Filled In the key column  N- (3,5- Dinitrobenzoyl ) -Leucine Phenylalkylamide  Optical division of derivatives

R k ₁ k ₂ α CH ₃ 4.18 4.75 1.13 CH ₃ CH ₂ 3.16 4.04 1.28 CH ₃ (CH ₂ ) ₂ 2.55 3.42 1.34 CH ₃ (CH ₂ ) ₃ 2.21 3.11 1.41 CH ₃ (CH ₂ ) ₄ 2.01 2.87 1.42 CH ₃ (CH ₂ ) ₅ 1.84 2.62 1.43 CH ₃ (CH ₂ ) ₉ 1.36 2.09 1.54

k _{1 ,} k _{2 ,} α are as defined in <Table 1>.

Claims (2)

Chiral stationary phase (CSP1) represented by the formula:

Where R is based on Sephacler, an antibiotic that is characterized by being hydrogen, one or two methyl groups, one or two methoxy groups, one or two t-butyl groups, one or two nitro groups, one or two chloro groups Chiral stationary phase. A chiral column filled with a chiral stationary phase (CSP 1) represented by Chemical Formula 1 according to claim 1.