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Scheme 21. Second-generation synthesis of boceprevir. D-DTTA-di-p-toluoyl-D-tartaric acid; L-DTTA-di-p-toluoyl-L-tartaric acid.
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Biocatalysis has become an attractive tool in modern synthetic chemistry both in academic and industrial settings, offering access to enantiopure molecules. In industry, biocatalysis found use in small molecule pharmaceutical development. For several amine-containing drugs, biotransformations were applied in the process routes, improving the origin...
Citations
... One of the enzyme activities for which this diversity is required is the (asymmetric) reductive amination of prochiral ketones with free ammonia. Indeed, the obtained (chiral) amines are found in many active compounds and in the most frequently used chemical intermediates for the production of pharmaceuticals and fine chemicals [14][15][16][17] . In addition to the successful application of transaminases in industry 18 , enzymes used for amine synthesis from ketones [19][20][21][22][23] include the Amine Dehydrogenases engineered from wild-type amino acid dehydrogenases (eng-AmDHs) 24,25 , native AmDHs (nat-AmDHs) 26 recently identified by our group, some reductive aminases (RedAms) 27,28 , a subclass of imine reductases (IREDs) active with ammonia, and engineered ɛ-deaminating L-lysine dehydrogenases 29 . ...
Native amine dehydrogenases offer sustainable access to chiral amines, so the search for scaffolds capable of converting more diverse carbonyl compounds is required to reach the full potential of this alternative to conventional synthetic reductive aminations. Here we report a multidisciplinary strategy combining bioinformatics, chemoinformatics and biocatalysis to extensively screen billions of sequences in silico and to efficiently find native amine dehydrogenases features using computational approaches. In this way, we achieve a comprehensive overview of the initial native amine dehydrogenase family, extending it from 2,011 to 17,959 sequences, and identify native amine dehydrogenases with non-reported substrate spectra, including hindered carbonyls and ethyl ketones, and accepting methylamine and cyclopropylamine as amine donor. We also present preliminary model-based structural information to inform the design of potential (R)-selective amine dehydrogenases, as native amine dehydrogenases are mostly (S)-selective. This integrated strategy paves the way for expanding the resource of other enzyme families and in highlighting enzymes with original features.
... [1,2,3,4,5] An estimated 40 % of active pharmaceutical ingredients (APIs) contain amine moieties. [6][7][8][9] Hence, efficient synthetic pathways for the production of such chiral building blocks are highly desired. Purely chemical approaches, such as reductive amination via metal catalysts, nucleophilic addition or diastereomeric crystallization of enantiomerically pure amine salts, [6] may face challenges due to harsh reaction conditions or a generally high amount of waste produced in the reaction caused by the use of protective groups or discarding of the undesired enantiomer. ...
... [6][7][8][9] Hence, efficient synthetic pathways for the production of such chiral building blocks are highly desired. Purely chemical approaches, such as reductive amination via metal catalysts, nucleophilic addition or diastereomeric crystallization of enantiomerically pure amine salts, [6] may face challenges due to harsh reaction conditions or a generally high amount of waste produced in the reaction caused by the use of protective groups or discarding of the undesired enantiomer. [1,2,4,5,10] Here, enzymatic synthesis approaches present a wellapplicable alternative to conventional chemical synthesis, allowing to circumvent some of the mentioned challenges. ...
... A spectrum of enzyme classes, depending on the desired reaction conditions, can be employed for amine production: amine and amino acid dehydrogenases, P450 monooxygenases and amine oxidases, imine reductases, lipases, Pictet-Spenglerases, berberine bridge enzymes and transaminases. [6][7][8][9] The latter are a well-established enzyme class of pyridoxal-5'phosphate-dependent (PLP-dependent) transferases, which transfer amino groups between an amine donor and an amine acceptor. This work will focus on amine transaminases (ATAs), a subgroup of ω-transaminases, which found a broad applicability in enantioselective amine synthesis, converting carbonyl moieties into amino groups without requiring the presence of a carboxylic group in the substrate. ...
This study explores a combination of the concept of enantioselective enzymatic synthesis of β‐chiral amines through transamination with in situ product crystallization (ISPC) to overcome product inhibition. Using 2‐phenylpropanal as a readily available and easily racemizing substrate of choice, (R)‐β‐methylphenethylamine ((R)‐2‐phenylpropan‐1‐amine) concentrations of up to 250 mM and enantiomeric excesses of up to 99 % are achieved when using a commercially available transaminase from Ruegeria pomeroyi in a fed‐batch based dynamic kinetic resolution reaction on preparative scale. The source of substrate decomposition during the reaction is also investigated and the resulting unwanted byproduct formation is successfully reduced to insignificant levels.
... Many chemo-enzymatic reactions for the synthesis of small-molecule active pharmaceutical ingredients (APIs) on large scale have been implemented. [8][9][10][11][12][13] Notable examples from the pharmaceutical industry include 1) the ketoreductase-mediated synthesis of a chiral diol intermediate of an antitumoral gamma secretase inhibitor; [14] 2) the chemoenzymatic processes involving transaminases to synthesize chiral amine precursors of antidiabetic sitagliptin [15] or antihypertensive sacubitril; [16] 3) the application of an imine reductase for reductive amination and consecutive kinetic amine resolution to yield an anticancer lysine-specific demethylase-1 inhibitor; [17] and 4) the comprehensive synthesis of anti-HIV islatravir through a series of reactions involving five evolved and four auxiliary enzymes. [18] In more recent years, enzymes like oxidases, transpeptidases and ligases have been utilized to access complex macromolecules such as modified peptides and oligonucleotides, as well as other biopolymers or bioconjugates with therapeutic applications. ...
Excelzyme, an enzyme engineering platform located at the Zurich University of Applied Sciences, is dedicated to accelerating the development of tailored biocatalysts for large-scale industrial applications. Leveraging automation and advanced computational techniques, including machine learning, efficient biocatalysts can be generated in short timeframes. Toward this goal, Excelzyme systematically selects suitable protein scaffolds as the foundation for constructing complex enzyme libraries, thereby enhancing sequence and structural biocatalyst diversity. Here, we describe applied workflows and technologies as well as an industrial case study that exemplifies the successful application of the workflow.
... Amines are among the most widely used compounds in the chemical and related industries, [1,2] as pharmacy, [3,4] agrochemistry [5] and rubber manufacturing. [6,7] On the one hand, many amines are toxic compounds and can be uncontrollably released into the environment as a result of the use of a number of dyes [8] and pesticides. ...
A new efficient approach to the synthesis of intensely colored 2‐(2‐oxo‐3H‐pyrrol‐3‐ylidene)malononitrile (OP) was developed. Using the reaction of OP with amines, a series of previously unknown colorless organic salts – dicyano(2‐oxo‐2,5‐dihydro‐1H‐pyrrol‐3‐yl)methanides were synthesized, bearing in the structure two fragments of same or diverse amines of different nature (covalent and ionic). TGA‐DTA study of the synthesized salts showed a thermal reversibility of their formation. The introduction of violet OP into paper followed by treatment with amine vapors revealed prospects of using such a system for naked‐eye detection of volatile amines. It was also shown that OP‐based rewritable paper can be created, to which a data can be applied by local heating and erased by the action of amine or temperature.
... Chiral amines have found extensive applications as building blocks in natural product synthesis, 1-7 pharmaceutical agents [8][9][10][11][12] and bioactive compounds. [13][14][15][16][17][18][19] According to Yang and co-authors, 20 approximately 35% of the top 200 small molecule drugs sold in 2020 contained at least one chiral amine subunit. ...
... [28][29][30][31][32][33][34][35][36] In this scenario, the search for newly effective chiral amines is a continuous process and has attracted attention from both academia and industry. Enantiomerically pure amines have been synthesized through several methods including reductive coupling of ketimines, 37 metal-catalized asymmetric hydrogenation, 7 biocatalysis, 12,[38][39][40] enantioselective reductive amination, 41 Mannich-type coupling, 42 diol diamination, 43 among many others. The Figure 1 shows examples of representative chiral amines and its applications. ...
In this work we described the synthesis and characterization of a series of novel chiral 1,5-diamines derived from (R)-(+)-camphor through simple procedures in moderate to good yields. These new enantiopure compounds constitute a new family of chiral diamines with potential applicability as chiral building blocks, bioactive products or chiral ligands for asymmetric transformations.
... Chiral amines are important structural motifs in biopharmaceuticals and fine chemicals, with 40%-45% of small-molecule drugs and industrial/agricultural fine chemicals containing these structures (Cabre et al., 2022;Ferrandi & Monti, 2017;Zawodny & Montgomery, 2022). Compared with chemical synthesis, biocatalytic preparation of chiral amines has the advantages of high stereoselectivity and environmental friendliness, so it is increasingly widely used in the asymmetric synthesis of chiral amines (Fuchs et al., 2015;Huo et al., 2020;Kelly et al., 2018). ...
Bioproduction of chiral amines is limited by low transaminase (TA) activity on nonnatural substrates, leading to the need for protein engineering. To address the challenge of quickly and precisely identifying the engineering targets, a strategy was proposed based on analyzing the mode changes in the high‐energy intermediate state (H‐state) of the substrate–enzyme complex during catalysis. By substituting the residues with minimal structural changes in catalytically active mode (A‐mode) and distance‐free mode (F‐mode) of the H‐state complex with more conserved ones to stabilize it, a TA mutant M5(T295C/L387A/V436A) with 121.9‐fold higher activity for synthesizing the (S)‐Rivastigmine precursor (S)‐1‐(3‐methoxyphenyl)ethylamine was created. The applicability of this strategy was also validated by engineering another TA for 1.52‐fold higher activity and >99% selectivity toward (R)‐3‐amino‐1‐butanol biopreparation. The much higher stereoselectivity of the mutant compared with the wild type (28.3%) demonstrated that this strategy is not only advantageous in engineering enzyme activity but also applicable for modulating stereoselectivity.
... In recent years, important progress has been made in biocatalytic ARA, and many enzymes capable of asymmetric reductive amination of carbonyl substrates have emerged. [51,52] These include NAD(P)H-dependent oxidoreductases such as amino acid dehydrogenases (AADHs) with amine dehydrogenase activity, amine dehydrogenases (AmDHs), engineered steroid dehydrogenases (OpDHs), and imine reductases (IREDs) with activity towards substrates containing carbonyl groups. [53] ...
This review summarizes the recent progress of organocatalytic and biocatalytic asymmetric reductive amination (ARA), a challenging but important topic for drug discovery and the pharmaceutical industry. At present, ARA can be divided into three categories: metal catalysis, organic catalysis, and biocatalysis. In the past decade, transition metal‐catalysed ARA has been well established. Organocatalytic ARA has emerged as a powerful alternative to metal‐catalysed ARA, the hydrogen sources used in organocatalytic ARA are usually Hantzsch esters, benzothiazolines, boranes, and hydrosilanes, which require Lewis base or phosphoric acid catalysts to activate them to give secondary chiral amines. It is worth mentioning that biocatalytic ARA has made remarkable progress in the last decade, amino acid dehydrogenases, amine dehydrogenases, opine dehydrogenases and imine reductases have been successfully used in ARA.
... (1, 16). Imines are compounds that are synthesized through compounds containing amine groups and have a wide range of biological activity properties such as antibacterial, antifungal, antimalarial, antituberculosis, antiviral, anti-inflammatory, antidiabetic, antipyretic, and enzyme inhibition (17,18). ...
In this study, new pyridine-based imine compounds were synthesized and it was investigated whether these compounds inhibit the D2 Dopamine receptor (6CM4) in silico. The structures of the compounds synthesized using the microwave method were determined by 1H-NMR, 13C-NMR and elemental analysis techniques. Later, Molecular Docking (MD) studies were used to determine the effects of synthesized compounds and risperidone on the D2 receptor. Risperidone drug and synthesized molecules (8-10) strongly interacted with D2 Dopamine Receptor /PDB: 6MC4 with energies of -10.34, -6.79, -6.95, -7.07 kcal/mol. The order of activity detected in the synthesized compounds is 10, 9, and 8 when the results are examined.
... There are different types of catalysts employed in chemical synthesis, but enzymes in different preparations (well as free biocatalysts, cells free extracts, or whole cells systems) have attracted great interest since the last years of the past century [26][27][28][29]. In the mid-1980s, the development of cloning and overexpression techniques, the advances in immobilization procedures, and the application of enzymes in non-aqueous systems have allowed the development of several enzymatic methodologies for the synthesis of high added-value compounds, including the preparation of APIs [30][31][32][33][34]. Biocatalyzed reactions usually occur with high efficiency, excellent selectivity, good yields, environmental sustainability, and lower costs, which make them more attractive from an industrial perspective. ...
Asymmetric oxidation processes have constituted a valuable tool for the synthesis of active pharmaceutical ingredients (APIs), especially for the preparation of optically active sulfoxides, compounds with interesting biological properties. Classical approaches for these oxidative procedures usually require the application of non-sustainable conditions that employ hazardous reagents and solvents. In the last decades, chemists have tried to combine the preparation of valuable compounds of high yields and selectivities with the development of more sustainable protocols. To achieve this objective, greener solvents, reagents, and catalysts are employed, together with the use of novel chemical techniques such as flow catalysis or photocatalysis. The last efforts in the development of greener approaches for the preparation of APIs and their intermediates using oxidative procedure will be reviewed herein. Most of these approaches refer to biocatalytic methods, in which mild reaction conditions and reagents are employed, but other novel techniques such as photocatalysis will be described.
... Enzyme applications in the large-scale industrial production of various chemicals have proven particularly advantageous in obtaining optically pure substances [1, 3,4]. Among optically active compounds, chiral amines are of great importance in the agrochemical and pharmaceutical industries [5][6][7]. Amines and amino compounds account for up to 90% of the top-selling or approved small molecular drugs, and around 30% of pesticides are chiral amine molecules [3]. ...
D-amino acids are valuable building blocks for the synthesis of biologically active compounds and pharmaceuticals. The asymmetric synthesis of chiral amino acids from prochiral ketones using stereoselective enzymes is a well-known but far from exhausted approach for large-scale production. Herein, we investigated a pyridoxal-5′-phosphate-dependent D-amino acid transaminase from Haliscomenobacter hydrossis as a potential biocatalyst for the enzymatic asymmetric synthesis of optically pure aliphatic and aromatic D-amino acids. We studied the catalytic efficiency and stereoselectivity of transaminase from H. hydrossis in the amination of aliphatic and aromatic α-keto acids, using D-glutamate as a source of the amino group. We constructed a one-pot three-enzyme system, which included transaminase and two auxiliary enzymes, hydroxyglutarate dehydrogenase, and glucose dehydrogenase, to produce D-amino acids with a product yield of 95–99% and an enantiomeric excess of more than 99%. We estimated the stability of the transaminase and the cofactor leakage under reaction conditions. It was found that a high concentration of α-keto acids as well as a low reaction temperature (30 °C) can reduce the cofactor leakage under reaction conditions. The obtained results demonstrated the efficiency of transaminase from H. hydrossis in the asymmetric synthesis of enantiomerically pure D-amino acids.
