Two-step desymmetrization of dipyrazolidyl 3-phenylglutarate via lipase-catalyzed hydrolysis in organic solvents

doi:10.1016/j.ces.2015.09.024

Chemical Engineering Science

Volume 139, 12 January 2016, Pages 41-48

https://doi.org/10.1016/j.ces.2015.09.024 Get rights and content

Highlights

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A sufficient condition for obtaining enantiomerically enriched product was proposed.
•
Dipyrazolidyl 3-phenylglutarate but not the diester was employed as the substrate.
•
Results agreed with the experimental data from the kinetic and thermodynamic analysis.

Abstract

A theoretical analysis on comparing the enzyme performance in a single-step desymmetrization, single-step kinetic resolution, and two-step desymmetrization (i.e. a single-step desymmetrization followed by a sequent kinetic resolution) is reported. On the basis of ee^* ≧ 0.95, a sufficient condition of E₃E₂⁻¹ ≧ 10 and E₁ ≧ 2 is proposed for obtaining an acceptable yield of X_2R^*>0.412 for the desired enantiomer in the two-step desymmetrization process, in comparison with E₁ ≧ 39 for the single-step desymmetrization and E₃E₂⁻¹ ≧ 20 for the single-step kinetic resolution. With the CALB-catalyzed hydrolytic desymmetrization of dipyrazolidyl 3-phenylglutarate (1) in MTBE as the model system, enantiomerically pure (R)-monopyrazolidyl 3-phenylglutarate ((R)-2) can then bfdee prepared. Moreover from the kinetic analysis, the best reaction condition of using 20% water-saturated MTBE as the medium at 45 °C is selected for improving the enzyme activity and stereoselectivity. The thermodynamic analysis also indicates that the enzyme stereo-discrimination in the desymmetrization and sequent kinetic resolution is mainly entropic-driven.

Graphical Abstract

Introduction

Lipases (E.C. 3.1.1.3) have been employed as the biocatalysts for synthesizing a variety of lipids, food and flavors, pharmaceuticals, fine chemicals, cosmetics, biodiesels, and polymers (Hasan et al., 2006, Kobayashi, 2009, Tan et al., 2010, Soumanou et al., 2013). A very useful feature of these enzymes is the enantiodiscrimination with which the preparation of single enantiomer of alcohol, acid, and amine is fulfilled (Bornscheuer and Kazlauskas, 2006, Ghanem, 2007, Kamal et al., 2008, Faber, 2011, Paravidino et al., 2012). Many important intermediates and building blocks for various synthetic applications belong to diol, diamine, dicarboxylic acid, or anhydride that contains one or more prochiral or chiral centers. When using a symmetrical prochiral or meso compound as the substrate, the first-step desymmetrization followed by a subsequent kinetic resolution can generally increase the optical purity, yet with the price of decreasing the yield of the required enantiomer (Bornscheuer and Kazlauskas, 2006, Faber, 2011). Quantitative analysis in enzyme-catalyzed two-step desymmetrization of prochiral or meso compounds is more complicated than that in a single-step desymmetrization or kinetic resolution, as at least two more kinetic parameters should be considered. This formulation remains still unclear and the relation of the theoretical to the experimental analysis has not been clearly discussed.

Enantiomerically pure 3-substituted glutarates are key structural elements of many drug intermediates and building blocks in organic synthesis (Fryszkowska et al., 2005, Gopinath et al., 2012, Jung et al., 2013). Enantiomerically enriched 3-arylglutarates have been prepared from alcoholic ring-opening of cyclic anhydrides via organocatalysts or enzymes (Chaubey et al., 2008, Park et al., 2010, Roy et al., 2014, Fryszkowska et al., 2006, García-Urdiales et al., 2011, Palomo and Cabrera, 2012, Liu et al., 2014) and enzyme-catalyzed hydrolysis, alcoholysis, aminolysis, or ammonolysis of dialkyl 3-arylglutarates (Yu et al., 2000; Homann et al., 2001; Lopez-Garcia et al., 2003; Cabrera et al., 2008; Wang et al., 2010a, Wang et al., 2010b; Cabrera and Palomo, 2011; Liu et al., 2012; Nojiri et al., 2013). In general it is difficult to develop an efficient desymmetrization process leading to high enantiomeric purity and yields for the desired enantiomer at the mild reaction condition after inspecting the experimental data reported in the references of this paragraph. It is not only about the reaction conditions but also about the biocatalysts.

As a part of our ongoing efforts toward using azolides as the substrate for preparing optically active compounds (Cheng et al., 2012, Tsai, 2015, Wang et al., 2010a, Wang et al., 2009, Wang et al., 2010b, Yu et al., 2000), we aimed to employ dipyrazolidyl 3-phenylglutarate (1) as the model substrate for preparing an enantiomer of high enantiomeric purity and yield via CALB-catalyzed hydrolysis in MTBE (Scheme 1). A thorough kinetic analysis is firstly performed for proposing a sufficient condition leading to the efficient desymmetrization process giving ee^* ≧ 0.95 and X_2R^*>0.412. From the kinetic analysis, the kinetic constants shown in Scheme 1 are estimated from experimental data for selecting the best reaction condition. The thermodynamic analysis is moreover addressed and elucidated.

Section snippets

Model development

By using an excess of water for the hydrolysis and assuming the substrate concentrations to be much lower than the Michaelis-Menten constants, an irreversible first-order kinetics for 1, (R)-2, and (S)-2 can be derived and solved for the time-course molar fractions as follows (Faber, 2011): $X_{1}^{*} = \exp (- t *)$ ${X_{2 R}}^{*} = \frac{E_{1} [\exp (- E_{2} t *) - \exp (- t *)]}{(1 + E_{1}) (1 - E_{2})}$ ${X_{2 S}}^{*} = \frac{\exp (- E_{3} t *) - \exp (- t *)}{(1 + E_{1}) (1 - E_{3})}$ $X_{3}^{*} = 1 - X_{1}^{*} - {X_{2 R}}^{*} - {X_{2 S}}^{*}$

The dimensionless parameters are defined as: t*=(k₁+k₂)t as dimensionless time, E₁=k₁k₂⁻¹ as

Materials

Novozym 435 (Candida antarctica lipase B (CALB) immobilized on acrylic resins, containing 1–2% (w/w) water and has 7000 PLU/g by using lauric acid and 1-propanol as substrates at 60 °C) was purchased from Novozymes (Bagsvaerd, Denmark). 1 PLU is the amount of enzyme activity which generates 1 μmol of propyl laurate per minute under the defined conditions. Other chemicals of analytical grade were commercially available: pyrazole and 1 H-benzotriazole from Acros (Geel, Belgium); thionyl chloride

Theoretical modeling

The sufficient condition for carrying out an effective desymmetrization process followed by a sequent kinetic resolution for obtaining (R)-2 is that the kinetic parameters should follow the order: k₁ ≧ k₂, k₁>k₃, and k₄>k₃ and hence E₁ ≧ 1, E₃E₂⁻¹>1, and E₂(E₁+1)E₁⁻¹<1 (or k₂ ≧ k₁, k₂ ≧ k₄, and k₃>k₄ and hence E₁ ≦ 1, E₃E₂⁻¹<1, and E₂(E₁+1)E₁⁻¹>1 if (S)-2 is desired). If the reaction condition can only lead to a single-step desymmetrization, results of E₁(E₁+1)⁻¹≧X_2R^* ≧ 0 and ee^*=(E₁-1)(E₁+1)⁻¹

Conclusions

The theoretical analysis for comparing the enzyme performances in a single-step desymmetrization, a single-step kinetic resolution, and the two-step desymmetrization for preparing chiral compounds is investigated, in which the dimensionless groups of E₁, E₃E₂⁻¹, and E₃(E₁+1)E₁⁻¹ are employed as the parameters. An effective two-step desymmetrization process should fulfill the sufficient condition of E₁ ≧ 1, E₃E₂⁻¹>1, and E₂(E₁+1)E₁⁻¹<1. Moreover in order to obtain an acceptable yield with ee^* ≧

Nomenclature

E₁

selectivity defined as k₁k₂⁻¹

E₂, E₃

dimensionless parameters defined as k₃(k₁+k₂)⁻¹and k₄(k₁+k₂)⁻¹, respectively

enantiomeric excess defined as (X_2R−X_2S)(X_2R+X_2S)⁻¹

K_m,1, K_m,2

Michaelis–Menten constants leading to (R)-2 and (S)-2 from 1, respectively (mM)

k₁, k_2,k₃, k₄

kinetic constants shown in Schemes 1 (h⁻¹)

(S)₀

initial concentration for dipyrazolide substrate (mM)

absolute temperature (K)

time (h)

dimensionless time defined as (k₁+k₂)t

V_2R, V_2S

initial rates for (R)-2 and (S)-2 from 1,

Acknowledgments

Financial supports of NSC 104-2221-E-182-058-MY2 from Ministry of Science and Technology are appreciated.

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Two-step desymmetrization of dipyrazolidyl 3-phenylglutarate via lipase-catalyzed hydrolysis in organic solvents

Highlights

Abstract

Graphical Abstract

Introduction

Section snippets

Model development

Materials

Theoretical modeling

Conclusions

Nomenclature

Acknowledgments

Fluid Phase Equilib.

Enzyme Microb. Technol.

J. Mol. Catal. B: Enzym.

Tetrahedron Asymmetr

Tetrahedron Asymmetry

Tetrahedron Asymmetry

Tetrahedron Asymmetry

Tetrahedron

Enzyme Microb. Technol.

Coord. Chem. Rev.

Proc. Biochem.

Tetrahedron Asymmetry

Biotechnol. Adv

Tetrahedron Lett.

Tetrahedron Lett.

Hydrolases in organic synthesis: regio- and stereoselective biotransformations