Enzymatic Acylation Procedure

The enzymatic acylation of natural polyhydroxylated compounds in ionic liquids ([bmim]BF4 and [bmimJPpG), and organic solvents was carried out in stirred flasks according to the procedure reported elsewhere [5, 6, 17]. 

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Kinetic optical resolution of racemic alcohols and carboxylic acids by enzymatic acyl transfer reactions has received enormous attention in recent years56. The enzymes generally employed are commercially available lipases and esterases, preferentially porcine liver esterase (PLE) or porcine pancreatic lipase (PPL). Lipases from microorganisms, such as Candida cylindracea, Rhizopus arrhizus or Chromobacterium viscosum, are also fairly common. A list of suitable enzymes is found in reference 57. Standard procedures are described in reference 58. Some examples of the resolution of racemic alcohols are given39. 

The synthesis of enantioenriched acyl chloride 54 was originally conducted using a modification of procedures described by Villieras and coworkers for the preparation of racemic alcohol 55 (Scheme 12). Thus, the synthesis commenced with the preparation of racemic allylic alcohol 55 from 2,5-dimethoxytetrahydrofuran. The alcohol was resolved enzymatically, following procedures reported by Ogasawara, to provide enantioenriched alcohol (S)-55 in 50% yield and 99% ee. Protection of the alcohol with the sterically encumbered ferf-butyl dimethylsilyl group allowed a diastereoselective copper(I)-mediated conjugate addition of methyl magnesium bromide to... 

Backvall and coworkers also reported a DYKAT of unsymmetrical 1,3-diols with one small and one large group [83]. The method makes use of (0 selective enzymatic acylation of the least sterically hindered alcohol ii) epimerization of a secondary alcohol and in) intramolecular acyl migration in a syn-l,3-diol monoacetate in a one-pot procedure. Shvo s catalyst 1 (4 mol%) was used for the epimerization. CALB was used for the enzymatic resolution along with isopropenyl acetate as the acyl donor. The products were obtained in moderate to high yields (53-73%) and with ee values exceeding 99% in all cases. Also, the diastereomeric ratios for the sy -diacetates were consistently high, typically >90% syn (19). 

The DKR procedure described above was improved by Meijer and coworkers in 2007 [87]. The protocol was improved both in terms of reaction time (26 h instead of 72 h) and the required amount of acyl donor (the excess acyl donor could be reduced to 1.1 equiv). This was accomplished using a more effective acyl donor isopropyl 2-methoxyacetate for the enzymatic acylation. CALB was used for the kinetic resolution, and the para-methoxyphenyl derivative of the Shvo catalyst was used for racemization (22). All the DKR reactions were performed under reduced pressure (750 mbar) to eliminate the isopropyl alcohol from the reaction mixture. The isopropyl alcohol can be oxidized to acetone, and the latter can in subsequent reaction steps form unwanted condensation products with the amine substrates. The revised protocol afforded the products with excellent selectivity (96-99% ee). The yields were slightly lower (56-80%) than those obtained with the Backvall protocol [86], mainly due to problems with purification. 

Despite its widespread application [31,32], the kinetic resolution has two major drawbacks (i) the maximum theoretical yield is 50% owing to the consumption of only one enantiomer, (ii) the separation of the product and the remaining starting material may be laborious. The separation is usually carried out by chromatography, which is inefficient on a large scale, and several alternative methods have been developed (Figure 6.2). For example, when a cyclic anhydride is the acyl donor in an esterification reaction, the water-soluble monoester monoacid is separable by extraction with an aqueous alkaline solution [33,34]. Also, fiuorous phase separation techniques have been combined with enzymatic kinetic resolutions [35]. To overcome the 50% yield limitation, one of the enantiomers may, in some cases, be racemized and resubmitted to the resolution procedure. 

Acyl nitroso compounds (3, Scheme 7.2) contain a nitroso group (-N=0) directly attached to a carbonyl carbon. Oxidation of an N-acyl hydroxylamine derivative provides the most direct method for the preparation of acyl C-nitroso compounds [10]. Treatment of hydroxamic acids, N-hydroxy carbamates or N-hydroxyureas with sodium periodate or tetra-alkyl ammonium periodate salts results in the formation of the corresponding acyl nitroso species (Scheme 7.2) [11-14]. Other oxidants including the Dess-Martin periodinane and both ruthenium (II) and iridium (I) based species efficiently convert N-acyl hydroxylamines to the corresponding acyl nitroso compounds [15-18]. The Swern oxidation also provides a useful alternative procedure for the oxidative preparation of acyl nitroso species [19]. Horseradish peroxidase (HRP) catalyzed oxidation of N-hydroxyurea with hydrogen peroxide forms an acyl nitroso species, which can be trapped with 1, 3-cyclohexanone, giving evidence of the formation of these species with enzymatic oxidants [20]. 

This is a simple procedure for the enzymatic resolution of a secondary amine. The acylating agent can be modified by altering the substitution on the phenol ring. This tuning of the reactivity and selectivity should allow other amines to be resolved using a similar approach. 

This process has many benefits in the context of green chemistry it involves two enzymatic steps, in a one-pot procedure, in water as solvent at ambient temperature. It has one shortcoming, however-penicillin acylase generally works well only with amines containing an aromatic moiety and poor enantioselectivities are often observed with simple aliphatic amines. Hence, for the easy-on/easy-off resolution of aliphatic amines a hybrid form was developed in which a hpase [Candida antarctica hpase B (CALB)] was used for the acylation step and peniciUin acylase for the deacylahon step [22]. The structure of the acyl donor was also optimized to combine a high enanhoselectivity in the first step with facile deacylation in the second step. It was found that pyridyl-3-acetic acid esters gave optimum results (see Scheme 6.8). 

An enzymatic procedure for amine resolution, employing acylation by C. antarctica lipase B and deacylation by penicillin G acylase, has been demonstrated by Ismail et al. (Figure 14.14) [20]. The acylase catalyzed deacylation provides a greener process than the standard chemical deacylation as a result of the elimination of the salt waste stream typically generated by deacylating under strongly alkaline condi tions. It is also more amenable to sensitive functional groups that are not stable under basic conditions. A drawback of this approach is that the amide hydrolysis step is... 

The enzymatic hydrolysis of N-acylamino acids has been known for a century and was first detected in aqueous kidney preparations 3. Based on the finding that this enzymatic hydrolysis proceeds enantiospecifically 2, Greenstein and coworkers developed a general and very attractive procedure for the resolution of a vast number of racemic N-acylated amino acids to the corresponding L-amino acids catalyzed by aminoacylase (E.C. 3.5.1.14) whereas the N-acetyl-D-amino acid does not react13 (Fig. 12.3-1). 

Hence resolution of enantiomers is necessary to obtain optically active L-amino acids. Among many resolution methods, the enzymatic method using mold aminoacylase developed by us proved to be one of the most advantageous procedures. An acyl-DL-amino acid is selectively hydrolyzed by aminoacylase to give L-amino acid and unhydrolyzed acyl-D-amino acid. 

Enzymatic desymmetrization of substituted 1,3-propanediols has been used as key step in the synthesis of y-butyrolactones by Itoh and coworkers, Scheme (8) [54],The diols 43 were treated with lipase PS (Pseudomenas sp.) in the presence of vinyl acetate as acyl donor to afford acetates 44 in excellent chemical yields and very high enantiomeric excesses (90-98%). These monoacetates were then converted into hydroxy nitriles 46 using a three step procedure. Tosylation of the hydroxyl group of 44, followed by treatment with potassium cyanide in dimethyl sulfoxide at 80°C gave the corresponding acetates 45. The acetoxy groups of 45 were finally hydrolysed with lithium hydroxide in a... 

The most prominent cellulose ester produced on the industrial scale is cellulose acetate. The reaction is usually performed with acetic anhydride and with sulfuric acid as a catalyst. To minimize heterogeneities, acetylation is allowed to run nearly to completion, and subsequently partial ester hydrolysis is initiated by the addition of water until a desirable solubility is achieved that corresponds to a DS of about 2.5. Such higher acyl homologues as propanoyl or butanoyl exhibit more thermoplastic properties. Many specialized esters such as chiral (-)-menthyloxyacetates, furan-2-carboxylates, or crown-ether-containing acylates have been prepared on the laboratory scale and characterized by NMR spectroscopy. Various procedures have been applied, using anhydrides and acyl chlorides as acylating agents in combination with such bases as pyridine, 4-dimethylaminopyridine (DMAP), or iV,iV -carbonyldi-imidazole. The substitution pattern of cellulose acetates has also been modified by postchemical enzymatic deacetylation. Cellulose 6-tosylates have been used as activated intermediates for nucleophihc substitution to afford 6-amino-6-deoxy, 6-deoxy, or 6-deoxy-6-halo-celluloses. ... 

The second task, resolution of synthetic D,L-amino acids has been solved by conversion of the neutral amino acids into real carboxyUc acids by acylation of the amino group, either by the benzoyl or the formyl residue. The D,L-acyl amino acids then formed diastereomeric salts with optically active bases, mostly alkaloids, which differed in their solubihty in various solvents, and so could be separated by recrystallization. This method is still in use, although enzymatic procedures, specific oxidation of the D-antipode in the presence of D-amino acid oxidase or enzymatic, stereospecific removal of iV-acyl residues from d,l-AT-acetyl-amino acids by acylase, are more convenient. Certainly, L-amino acids became accessible from nature by the ester method, but without synthetic material, extended experiments of peptide couphng would have been impossible. 

Vedejs and Chen [39] described an efficient non-enzymatic system able to approach the efficiency of some of the lipase methods in enantioselectivity. The reaction was carried out in a 2 1 ratio racemic secondary alcohol acylating agent, in contrast to Evans procedure. The pyridinium salt 8 was prepared by reaction of the chiral 4-dimethylaminopyridine (DMAP) 6 with the commercially available chloroformate 7. This pyridinium salt proved to be unreactive to secondary alcohols. The reactivity was found only upon strict experimental conditions addition of a Lewis acid, then the racemic alcohol, followed by addition of a tertiary amine gave the carbonate 9. Under these conditions (using MgBr2 and triethylamine), (2-naphthyl)- -ethanol was converted (room temperature, 20 h and 54% conversion) into the (S)-carbonate (82% ee). The recovered alcohol showed 83% ee, revealing a stereoselectivity s=39 for the process. A number of 1-arylalkanols have been resolved by this procedure in 20-44% yield (based on the racemic material) and 80-94% ee. For the use of this system in enantiodivergent reactions, see Schemes 6.1 and 6.32. 

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