Bavo Vandekerckhovea,
Stefan Desimpela,
Bart Ruttensb,
Massimo Bocus
c,
Wim Temmermanc,
Bert Metten
b,
Veronique Van Speybroeck
c,
Thomas S. A. Heugebaert
a and
Christian V. Stevens
*a
aSynBioC Research Group, Department of Green Chemistry and Technology, Faculty of Bioscience Engineering, Ghent University, Coupure Links 653, 9000 Ghent, Belgium. E-mail: Chris.Stevens@UGent.be
bAjinomoto Omnichem, Cooppallaan 91, 9230 Wetteren, Belgium
cCenter for Molecular Modeling, Tech Lane Ghent Science Park, Campus Ardoyen, Ghent University, Technologiepark 46, 9052 Zwijnaarde, Belgium
First published on 13th May 2025
Amides play a crucial role in the pharmaceutical, animal health and agrochemical industry. Despite the availability of various catalytic systems and coupling reagents, many methods suffered from long reaction times and poor atom economy. The direct synthesis of primary amides remained particularly challenging due to the limited availability of suitable nitrogen sources. In this study, continuous flow technology was explored as a process-intensification approach for the direct amidation of methyl esters to produce primary amides. Methanolic ammonia was employed as a nitrogen source to enhance process efficiency while circumventing the limitations of aqueous ammonia and the hazards of gaseous ammonia. Seventeen substrates were screened to assess their aminolysis reactivity under these conditions. As a proof of concept, methyl picolinate was selected for continuous flow optimization using Bayesian optimization. Therefore, a custom-designed high-pressure, high-temperature continuous flow reactor was utilized to achieve efficient, safe and scalable synthesis (200 °C, 50 bar).
Extensive literature addresses the reactivity of carboxylic acids and esters, yet the direct synthesis of corresponding primary amides on a practical scale remains a persistent challenge. Various catalytic systems and coupling reagents have been investigated for converting carboxylic acids directly into amides, including DCC, DIC, EDC, HATU, HBTU and phosphonium salts. However, these typically complicate reaction work-up, substantially increase synthesis costs (including raw material expenses, recovery processes, regulatory compliance, and waste management), and lead to poor atom economy.3 An alternative approach involves the reaction of esters with ammonia or amines to produce the corresponding amides, with some catalysts being reported for the batch aminolysis of esters using ammonia in solution.4–6 Unfortunately, these methods often require harsh conditions, involve high costs, and are characterized by very long reaction times, often extending to several days. Given these limitations, the development of innovative and efficient approaches to amide synthesis is attracting growing interest.
The use of ammonia in organic synthesis poses several challenges, primarily stemming from its limited solubility in various solvents and the difficulties associated with handling its reactive and toxic gaseous form. In stirred tank reactors, gaseous ammonia often accumulates in the headspace above the solution, which reduces its reactivity and necessitates the use of excessive amounts to achieve desired outcomes. Commercially, ammonia is typically available either as an aqueous solution or dissolved in methanol. Although, aqueous solutions are the most sustainable, the hydrolysis caused by water is not compatible with many chemical processes and functional groups.
Continuous flow technology has been extensively employed as a process intensification strategy to mitigate environmental challenges of batch processes and to develop green, scalable protocols.7–9 This study aims to investigate the use of continuous flow reactors as a process intensification technology for the amidation of methyl esters to produce primary amides. Flow chemistry presents key advantages, such as the safe handling of hazardous materials and the efficient utilization of gases.10 The objective is to drastically reduce reaction times from days to minutes while minimizing the amount of ammonia required, all without relying on coupling reagents, additives or catalysts.11
Recently, Bayesian optimization (BO) has emerged as a powerful tool for the optimization of process parameters in continuous flow chemistry, offering an efficient strategy to navigate complex, multidimensional parameter spaces with minimal experimental effort. By constructing a probabilistic model of the objective function, BO balances exploration and exploitation, enabling rapid identification of optimal conditions.12 Numerous applications have demonstrated its effectiveness in fine-tuning reaction parameters, leading to significant improvements in yield, selectivity and process robustness.13 Here, Bayesian optimization will be employed to determine the optimal reaction conditions for this chemistry.
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Fig. 2 Direct amidation of methyl L-prolinate hydrochloride 1a (non-aromatic) and methyl picolinate 1b (aromatic) as non-aromatic and aromatic proof-of-concept substrates. |
Entry | NH3 eq.a | Time (min) | Temp. (°C) | Conversion 1ab (%) | 2a/–OH/diketob (%) | Conversion 1bb (%) | 2b/–OHb (%) |
---|---|---|---|---|---|---|---|
a A 7 N solution of NH3 in MeOH is used.b Conversion and NMR yield calculated relative to internal standard (dimethyl sulfone).c Reagent concentration remained constant. | |||||||
1 | 12.0 | 30 | 50 | 42 | 26/6/0 | 85 | 56/0 |
2 | 12.0 | 30 | 80 | 75 | 43/10/1 | 85 | 75/0 |
3 | 12.0 | 30 | 100 | 83 | 64/3/0 | 89 | 82/0 |
4 | 12.0 | 15 | 110 | 76 | 62/4/0 | 96 | 86/0 |
5 | 12.0 | 15 | 120 | 82 | 61/4/0 | 62 | 51/0 |
6 | 12.0 | 10 | 120 | 75 | 22/3/14 | 50 | 48/0 |
7 | 12.0 | 15 | 130 | 83 | 70/7/0 | 50 | 45/0 |
8 | 12.0 | 10 | 130 | 69 | 53/8/0 | 54 | 51/0 |
9c | 9.0 | 10 | 130 | 70 | 40/5/0 | 33 | 26/0 |
10c | 6.0 | 10 | 130 | 59 | 22/5/1 | 33 | 28/0 |
11c | 3.0 | 10 | 130 | 41 | 14/13/3 | 22 | 21/0 |
As mentioned before, the formation of both a hydrolysis byproduct and diketopiperazine was observed during the amidation of methyl L-prolinate hydrochloride (Fig. 2). To improve the selectivity of the aminolysis reaction towards the desired product, controlling the reagent concentration to suppress dimerization side reactions, along with careful optimization of reaction time and temperature, could further enhance amide formation. Especially, minimizing the presence of water is crucial to reduce hydrolysis. Karl–Fisher analysis of the 7 N ammonia solution in methanol confirmed that the solvent was ultra-dry, with a water content of 2.4 ppm. A reaction mixture was then prepared using methyl L-prolinate hydrochloride and the ammonia solution, stirred for 1 hour, and its water content was measured. This resulted in 1808 ppm (0.1808%, considered very wet) and these levels could interfere with moisture-sensitive reactions or promote unwanted side reactions. This corresponds with a water content of 0.18 equivalents relative to the methyl L-prolinate hydrochloride, making hydrolysis levels of up to 13% quite likely. The same procedure for methyl picolinate 1b resulted in a water content of 466 ppm (0.0466% or 0.045 eq.), which is acceptable for most standard reactions. The extent of hydrolysis observed during the aminolysis of methyl L-prolinate hydrochloride can likely be attributed to its hygroscopic nature. Therefore, various drying methods were evaluated to reduce possible hydrolysis (Table 2). Performing the reaction on a larger scale (×3) did not significantly reduce the formation of the hydrolysis side product, thus excluding glassware adsorbed water. The addition of trimethyl orthoformate (TMO) as an organic drying agent resulted in the formation of additional side products, while oven-dried methyl L-prolinate hydrochloride exhibited some degradation. As expected, using ammonia solution dried on molecular sieves did not effectively reduce hydrolysis to any notable extent either.
Entry | NH3 eq.a | Time (min) | Temp. (°C) | Attempt | Conversion 1ac (%) | 2a/–OH/diketoc (%) |
---|---|---|---|---|---|---|
a A 7 N solution of NH3 in MeOH is used.b 450 mg scale instead of 150 mg.c Conversion and NMR yield calculated relative to internal standard (dimethyl sulfone). | ||||||
1 | 12.0 | 10 | 130 | Original | 69 | 53/8/0 |
2b | 12.0 | 10 | 130 | Scale ×3 | 86 | 65/6/1 |
3 | 12.0 | 10 | 130 | Oven dried | 59 | 32/14/2 |
4 | 12.0 | 10 | 130 | TMO (0.5 eq.) | 95 | 14/17/2 |
5 | 12.0 | 10 | 130 | MS dried solvent | 72 | 51/10/1 |
To further explore the scope of methyl ester aminolysis in the formation of primary amides, several alternative substrates with a particular focus on heterocyclic structures, were evaluated at suboptimal conditions (130 °C, 10 minutes, Fig. 3). Eight additional substrates demonstrated moderate to excellent reactivity in forming the primary amide. Notably, pyrimidine 2g and pyrazine 2h exhibited remarkably high NMR yields. While the aminolysis of these compounds has been previously reported in literature, the reactions typically required prolonged times ranging from 1 to 3 days.16,17
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Fig. 3 NMR yield for the ammonia-mediated aminolysis of different substrates (amethyl ester hydrochlorides are used). |
Some substrates exhibited yields that could not be readily rationalized based on chemical intuition. To study the lack of reactivity observed for the other substrates, additional first-principles simulations were conducted.18,19 The problem was simplified by considering the 0 K electronic energy difference between the most stable conformers that could be identified for reactants and products. Simulations were performed both in the gas phase and with implicit methanol solvation. We found that the reaction energy, while straightforward, is a relatively good reactivity descriptor for the amidation reaction. The results, presented in Fig. 4, indicate an inverse correlation with the experimental yield. A more negative reaction energy indicates a greater thermodynamic driving force, suggesting that the reaction should be more favorable. This suggest that relative thermodynamic stability plays an important role in driving the reaction. The inclusion of implicit solvation effects and temperature corrections via reaction free energies did not significantly alter these trends. It must be noted that these calculations provide thermodynamic insights only and do not account for reaction kinetics (see ESI† for details). It is expected that, at high temperatures and in the presence of a protic solvent, specific intermolecular interactions and the hydrogen bonding patterns will play a major role during the reaction from a kinetic point of view. However to perform such extensive kinetic analysis from theoretical point of view, it would be necessary to model the reaction in explicit solvation and with molecular dynamics simulations, which is beyond the scope of the current study.
We hypothesize that an important structural parameter influencing the reaction yield is the ability of the amide to form an intramolecular hydrogen bond with an available electron donor. This could involve a nitrogen atom with a lone pair positioned ortho to the amide group. Such interactions may contribute to the inverse correlation observed between reaction energies and experimental yields. From a kinetic point of view, it could also facilitate favorable interactions between ammonia and the carbonyl group, potentially accelerating the reaction rate (analogue to coupling reagents e.g. HATU; kinetic study beyond scope of current work). The optimized geometries from the conformational search (Fig. 5) confirm that the observed stabilization of the amide is relatively well explained by the formation of such intramolecular hydrogen bond between the amide NH2 group and an electron-rich atom in the ortho position. Among the studied compounds, only two, 2e and 2m (highlighted in orange in Fig. 5), exhibit this intramolecular hydrogen bond while still having an experimental yield of zero. In the case of 2e, the only available hydrogen bond acceptor is the nitrogen atom of the pyrazole linker. However, its proximity to a second nitrogen atom reduces its electron density, making it a weaker hydrogen bond acceptor. For 2m, additional factors due to its relatively large size may contribute to the observed lack of yield, for example the steric repulsion of the isopropyl and benzyl groups.
Entry | NH3 eq. | Time (min) | Temp. (°C) | Conversion 1b (%) | NMR yield 2b (%) |
---|---|---|---|---|---|
1 | 8.9 | 5.5 | 159 | 54 | 26 |
2 | 8.0 | 0.7 | 182 | 39 | 13 |
3 | 5.7 | 9.8 | 197 | 53 | 27 |
4 | 4.2 | 4.9 | 136 | 22 | 12 |
5 | 10.2 | 7.9 | 146 | 62 | 41 |
6 | 2.0 | 2.2 | 165 | 20 | 4 |
7 | 12.0 | 10.0 | 188 | 80 | 59 |
8 | 20.0 | 20.0 | 188 | 89 | 58 |
9 | 14.7 | 15.1 | 199 | 85 | 53 |
10 | 20.0 | 9.7 | 197 | 75 | 47 |
11 | 13.3 | 10.9 | 194 | 79 | 57 |
12 | 11.6 | 20.0 | 194 | 92 | 53 |
13 | 17.7 | 30.0 | 200 | 97 | 69 |
14 | 20.0 | 30.0 | 139 | 98 | 70 |
Due to its ease of analysis and reliability compared to methyl L-prolinate, methyl picolinate 1b was selected for optimizing the flow process using Bayesian optimization. The primary objective was to simultaneously maximize both the conversion and the NMR yield of the target product, picolinamide. While these objectives are obviously correlated, particularly in the absence of competing reactions or degradation pathways, both objectives are used to identify high-yielding conditions. Although minimizing the equivalents of ammonia was considered as a potential objective, it was not incorporated into the optimization strategy. Chemical intuition suggests that higher ammonia equivalents would be favored due to ammonia's relatively unreactive nature. It was therefore anticipated that explicitly minimizing ammonia equivalents within the Bayesian optimization framework would lead to significantly suboptimal yields (Fig. 7).
Initially, the parameter boundaries were set as follows: NH3 equivalents (1–12 eq.), reaction time (0.5–10 min) and reaction temperature (130–200 °C) (entries 1–6). However, the algorithm's first suggestion after the initialization set (entry 7) already reached the maximum for two out of three parameters. To expand the optimization space, the boundaries for reaction time and ammonia equivalents were expanded to 20 minutes and 20 equivalents, respectively, demonstrating the flexibility of this approach. When boundaries are extended during optimization, the algorithm exhibits high uncertainty within the newly expanded process window. As a result, the next experiment (entry 8) reached the new maximum limits to explore this region. Afterwards, conversion gradually increased over subsequent iterations. Since entry 12 achieved near-full conversion but once again approached the parameter boundaries, the process window was expanded one final time, extending the reaction time to a maximum of 30 minutes. It is important to note that when ammonia equivalents exceed 12, the concentration of methyl picolinate had to be decreased due to the limitation of using a commercially available 7 N methanolic ammonia solution. Finally, nearly complete conversion was achieved using 20 equivalents of ammonia, a reaction time of 30 minutes and a temperature of 139 °C. Further application of the algorithm did not lead to any improvements. Thus, the optimal conditions were established after 8 additional experiments, next to the initial 6 screening experiments (Fig. 7). It is estimated that the algorithm would have achieved convergence faster if more suitable boundaries had been chosen from the start (based on technical feasibilities of equipment). Initially, it was hypothesized that eliminating headspace and using continuous flow could enhance the reactivity of dissolved ammonia gas, potentially reducing the required excess of reagent. However, Bayesian Optimization revealed that achieving desirable yields still necessitates a high excess of ammonia, even within a continuous flow setup. In addition, NMR analysis shows no byproduct formation, suggesting that the difference between conversion and yield may be attributed to unknown volatile byproducts.
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Fig. 8 Conversion, NMR and isolated yield for the ammonia-mediated aminolysis of different substrates in continuous flow (amethyl ester hydrochlorides are used). |
After optimization of the reaction parameters within the continuous flow setup, these substrates showed improved conversion rates and NMR yields compared to the non-optimal batch conditions (except 2i). Notably, no precipitation was observed for substrates utilizing hydrochloride salts as starting materials during the reaction or sample collection (2a and 2i). Final purification of all substrates was achieved via recrystallization (see ESI† for details).
Productivity (flow) = 0.00034 mol mL−1 × 1.333 mL min−1 × 60% yield × 122.1 g mol−1 = 0.033 g min−1 = 1.99 g h−1 | (1) |
In alignment with the principles of green chemistry, the presented protocol operates without the use of additives, catalysts, or stoichiometric amounts of coupling reagents, thereby minimizing waste generation and simplifying purification processes. Reaction times could be shortened from several hours down to 30 minutes, although high equivalents of ammonia are still necessary. Furthermore, scale-up can be readily achieved through continuous production compared to previously reported batch protocols, while simultaneously mitigating the hazards associated with the handling of ammonia gas at larger scales. Particular emphasis was set on developing a broadly applicable setup and protocol, leading to the decision to employ methanolic ammonia exclusively (acknowledged as green solvent). This decision also accounts for potential challenges related to hydrolysis or the presence of water-sensitive functional groups in other substrates (beyond those investigated in this study). Potential solubility limitations of substrates in methanol (because of this decision) could still be effectively managed due to the solid-handling capabilities of the system.
Footnote |
† Electronic supplementary information (ESI) available: A detailed description of the experimental work is provided in the ESI. See DOI: https://doi.org/10.1039/d5re00163c |
This journal is © The Royal Society of Chemistry 2025 |