Ali
Alipour Najmi
a,
Zhangping
Xiao
b,
Rainer
Bischoff
a,
Frank J.
Dekker
b and
Hjalmar P.
Permentier
*a
aDepartment of Analytical Biochemistry, Groningen Research Institute of Pharmacy, University of Groningen, A Deusinglaan 1, 9713 AV Groningen, The Netherlands. E-mail: h.p.permentier@rug.nl
bChemical and Pharmaceutical Biology, Groningen Research Institute of Pharmacy, University of Groningen, A Deusinglaan 1, 9713 AV Groningen, The Netherlands
First published on 9th September 2020
A practical, efficient, and selective electrochemical N-demethylation method of tropane alkaloids to their nortropane derivatives is described. Nortropanes, such as noratropine and norscopolamine, are important intermediates for the semi-synthesis of the medicines ipratropium or oxitropium bromide, respectively. Synthesis was performed in a simple home-made electrochemical batch cell using a porous glassy carbon electrode. The reaction proceeds at room temperature in one step in a mixture of ethanol or methanol and water. The method avoids hazardous oxidizing agents such as H2O2 or m-chloroperbenzoic acid (m-CPBA), toxic solvents such as chloroform, as well as metal-based catalysts. Various key parameters were investigated in electrochemical batch or flow cells, and the optimized conditions were used in batch and flow-cells at gram scale to synthesize noratropine in high yield and purity using a convenient liquid–liquid extraction method without any need for chromatographic purification. Mechanistic studies showed that the electrochemical N-demethylation proceeds by the formation of an iminium intermediate which is converted by water as the nucleophile. The optimized method was further applied to scopolamine, cocaine, benzatropine, homatropine and tropacocaine, showing that this is a generic way of N-demethylating tropane alkaloids to synthesize valuable precursors for pharmaceutical products.
Despite the importance of selective N-demethylation of tropane alkaloids, it has remained a challenging step for synthetic chemists.8 Different approaches have been reported for N-demethylation of the tertiary amine in atropine and scopolamine using agents such as 2,2,2-trichloroethyl chloroformate,9 α-chloroethyl chloroformate,10 KMnO410,11 and photochemistry.12 Although these methods provide low to excellent yields (16–100%) of 5 and 8, they use toxic solvents and chemicals producing hazardous waste and by-products.
The original Polonovski reaction has proven to be effective for the N-demethylation of tertiary N-methylamines.13 Over the past decade, modifications of the reaction based on iron catalysts have been developed for the N-demethylation of tropane and opiate alkaloids.2,3,6,8,14–19 These reactions are based on a multi-step process, comprising the oxidation of the tertiary amine using m-chloroperbenzoic acid (m-CPBA) or H2O2 to the corresponding N-oxide, which is then isolated as the HCl salt. The N-oxide salt is N-demethylated with iron-based catalysts such as FeSO4·7H2O, FeCl2·4H2O, or Fe(NH4SO4)2,14,15 iron porphyrin complex,16,17 ferrocene,8 and iron powder2 which need to be separated from the reaction mixture by chelating with EDTA or TPPS (meso-tetra(4-sulfophenyl)porphyrin)14,15 or by filtration.16–18
Recent works have focused on using greener solvents such as ethanol and isopropanol, instead of chloroform, in the N-demethylation of N-oxides.3,6,18,20 In one approach, iron nanoparticles have been studied as catalyst for the N-dealkylation of various alkaloids reporting a yield of 85% for the synthesis of noratropine.18 However, chloroform is still in use for N-oxide formation in the first step of the reaction.18 Do Pham et al. studied the N-demethylation of atropine and scopolamine using Fe(III)–TAML (tetra-amidato macrocyclic ligand) as catalyst in ethanol without isolation of the N-oxide as the hydrochloride salt, reporting a yield of about 80% for noratropine and norscopolamine.3,6 However, this method uses 50 equivalents of H2O2, which must be deactivated at the end of the process by treatment with MnO2.3,6 A detailed comparison of these methods with respect to their advantages and disadvantages is provided in Table S1.†
Previous methods use either toxic organic solvents, require powerful oxidants or necessitate chromatographic purification or filtration to remove catalysts, all of which add to the overall cost of the procedure and increase the impact on the environment notably for large-scale production processes.2,3,6,8,14–19
Concerted N-demethylation/N-acylation strategies based on palladium catalysts have been reported for atropine and opiate alkaloids.21–24 Although these methods have been applied for the synthesis of the semi-synthetic opiates naltrexone22 and buprenorphine,23 the reaction is not selective for atropine, since the hydroxyl group is dehydrated, leading to the formation of apoatropine.21 In addition, application of palladium catalysts in commercial medicines leads to cost issues due to the need for palladium removal below the required level of 10 ppm.3
Organic electrosynthesis has several advantages over traditional methods, such as the limited use of hazardous chemicals, mild reaction conditions, a simple system design, scalability and notably sustainability.25–27 It is thus considered to be a “green” alternative to widely used organic synthesis methods.28,29 While electrochemistry has been applied for the oxidation of aliphatic amines,30–32 ferrocene-mediated oxidation of cyclohexylamines,33,34 and some phenethylamines35 as well as the synthesis of a wide-range of organic molecules (for a detailed list see a recent review36), there is no report regarding its application for the N-demethylation of bicyclic tertiary amines in general and tropane alkaloids in particular. Recently, Gul et al. reported optimized conditions for generating N-dealkylated lidocaine.37 In another study, electrochemical N-dealkylation of fesoterodine was reported.38 However, the reported concentrations (0.1–1 mM) are far lower than what is used in practical synthesis procedures, while only achieving less than 30% conversion to the N-dealkylated drugs.37,38
In order to overcome the challenges of the available organic synthesis methods, we developed a generic electrochemical N-demethylation strategy for tropane alkaloids that gives both good selectivity and yields in gram-scale synthesis. Product isolation is straightforward using liquid–liquid extraction. The reaction proceeds at room temperature in one step without the need for metal-based catalysts in an ethanol or methanol/water mixture. After having established the electrochemical synthesis reaction, we studied the mechanistic pathway showing that this reaction proceeds via the formation of an iminium intermediate that reacts with water to give nortropanes.
All electrochemical measurements were performed with an Autolab potentiostat (Metrohm AG, Herisau, Switzerland) using NOVA software (Metrohm AG) at room temperature under ambient atmosphere. Between each experiment, electrode surfaces were washed with water and ethanol and then dried under atmospheric conditions for both the μ-PrepCell and home-made cells.
The final organic extract was dried (Na2SO4), filtered and the solvent removed under vacuum to give the product. The yield was determined from the weight and 1HNMR of the isolated product.
As there is only one methyl group difference between atropine and noratropine, separation of these two compounds from any reaction mixture can be challenging. For this reason, it is important to have full conversion at the end of the reaction. Using only ethanol as solvent did not result in full conversion. Increasing the reaction time and decreasing the applied current either did not lead to complete conversion or resulted in overoxidation, a decreased yield and a complex reaction mixture of byproducts, as determined by LC–MS (Table 2, entries 1–5). Although increasing the applied current led to full conversion, analysis of the final reaction mixture by LC–MS showed a more complex reaction mixture including different overoxidized byproducts (see LC–MS chromatogram in Fig. S5†). We then considered whether addition of water to the reaction mixture would result in full conversion, as the electrochemical N-demethylation of atropine is similar to the Shono-type electrochemical oxidations, which have been applied for the activation and functionalization of C–H bonds adjacent to nitrogen atoms.41 It has been hypothesized that water or methanol may act as a nucleophile trapping the iminium intermediate produced during the electrochemical reaction.28,42,43 Increasing the water content of the reaction mixture led to full conversion and decreased the required time and charge for completion of the reaction (Table 2, entries 6–8), while giving a moderate yield of noratropine (44–67%). LC–MS analysis of the final reaction mixture still showed multiple byproducts (Table 2, entry 8). Reducing the applied current to 4 mA resulted in 82% isolated yield after silica-gel chromatography and a less complex final reaction mixture with only two minor byproducts (Table 2, entry 9; LC–MS chromatogram Fig. S6a†).
Entry | Current/time | Added water | Conversionb (%) | Yield (%) |
---|---|---|---|---|
a Absolute ethanol was used as solvent and the residual amount of water was not measured. b Determined by LC–MS. c Isolated yield (based on weight and 1HNMR) after silica-gel chromatography. d Isolated yield (based on weight and 1HNMR) after 3-step LLE. e Isolated yield (based on weight and 1HNMR) after 2-step-NaOH-LLE. f Isolated yield (based on weight and 1HNMR) after 2-step-NH4OH-LLE. | ||||
1 | 6 mA/3 h | 0a | 90 | 70b |
2 | 6 mA/5 h | 0a | 90 | 37b |
3 | 6 mA/7 h | 0a | 93 | 28b |
4 | 4 mA/6 h | 0a | 88 | 25b |
5 | 8 mA/3 h | 0a | 99 | 52b |
6 | 6 mA/5 h | 2.2 M (4% v/v) | 92 | 44b |
7 | 6 mA/5 h | 11 M (20% v/v) | 99 | 65b |
8 | 6 mA/3.5 h | 18.5 M (33% v/v) | 99 | 67b |
9 | 4 mA/4.5 h | 18.5 M (33% v/v) | 99 | 82c |
10 | 4 mA/4.5 h | 18.5 M (33% v/v) | 99 | 84d |
11 | 4 mA/4.5 h | 18.5 M (33% v/v) | 99 | 68e |
12 | 4 mA/4.5 h | 18.5 M (33% v/v) | 99 | 80f |
Considering the low complexity of the final reaction mixture, based on LC–MS analysis, we tested whether the desired product could be purified by liquid–liquid extraction (LLE). As waste production and solvent consumption at the final purification step of active pharmaceutical ingredients are responsible for more than half of the overall manufacturing expenses, there is a strong incentive to adopt other purification methods than chromatography.44,45 Do Pham et al. applied a three step LLE approach to isolate noratropine and norscopolamine from their final reaction mixture with high purity.3,6 Three different LLE techniques were evaluated and compared to chromatographic purification to isolate noratropine from the final reaction mixture obtained under the same reaction conditions showing that comparable yields to chromatographic purification can be obtained by three- or two-step LLE (Table 2, entries 10–12, LC–MS chromatogram of final reaction mixture Fig. S6b–d†). Another four replicates of the final reaction condition (Table 2, entries 9–12) were performed with another set of electrodes and reported in Table S2.†
In order to investigate the sensitivity of the electrochemical N-demethylation reaction to either solvent or supporting electrolyte, methanol was selected as an alternative solvent and four different supporting electrolytes were tested. As listed in Table 3, the electrochemical N-demethylation of atropine proceeded with good yield under all of these reaction conditions (LC–MS chromatogram of final reaction mixture Fig. S7†). However, using LiBr as supporting electrolyte required a longer reaction time to complete the conversion of atropine as compared to other supporting electrolytes.
Having studied different reaction conditions for the electrochemical N-demethylation of atropine, we investigated the generality of this approach by applying it to other tropane compounds (Table 4). We performed electrochemical N-demethylation of tropacocaine 10 in the home-made batch cell resulting in an isolated yield of 70% of nortropacocaine. It is noteworthy that performing the electrochemical reaction using its salt form (tropacocaine·HCl) did not lead to any conversion, possibly due to the acidic pH of the reaction solution with tropacocaine·HCl. Adding an equimolar of sodium carbonate to the reaction solution increased the pH of the solution to about 10 leading to 95% conversion (Table S3†). The facile N-demethylation reaction at basic pH is likely due to the fact that the lone–pair electrons of the tertiary amine group are more easily available for abstraction than under acidic conditions, when the amine is largely protonated.37,38,46,47 Besides tropane alkaloids, we have applied the electrochemical N-demethylation reaction to other examples of organic compounds having N-methylpiperidine or N-methylpyrrolidine in their chemical structures (Table S4†). Although the reaction proceeds with those compounds as well, the isolated yield of the secondary amine is lower than for the nortropanes, especially for N-methylpyrrolidine; the drug clemastine 14 was mostly converted to a product which was hydroxylated on the carbon adjacent to the nitrogen.
Tropane alkaloids | Synthesized nortropanes | ||
---|---|---|---|
a 0.16 mmole. b 0.2 mmole. c Isolated yield after 2-step-NH4OH-LLE. d Isolated yield after silica-gel chromatography. e Electrochemical oxidation potential versus Ag/AgCl reference electrode (CV curves in Fig. S9†). f Faradaic efficiency (2-electron oxidation). | |||
Scopolamine 2b |
![]() |
83%c |
![]() |
1.01 Ve | 4 mA/3 h | ||
74%f | |||
Cocaine 3b |
![]() |
60%d |
![]() |
1.04 Ve | 4 mA/3 h | ||
53%f | |||
Tropacocaine 10a |
![]() |
73%c |
![]() |
1.00 Ve | 4 mA/7 h | ||
22%f | |||
Homatropine 11b |
![]() |
63%c |
![]() |
0.97 Ve | 4 mA/7 h | ||
24%f | |||
Benzatropine 12a |
![]() |
72%d |
![]() |
0.91 Ve | 4 mA/12 h | ||
12%f |
Having studied the reaction conditions, the electrochemical N-demethylation of atropine was performed at the gram-scale using a stack of electrodes in a batch cell (Fig. S3†). As listed in Table 5, good yields of noratropine were obtained by a three-step LLE method after electrochemical reaction, with high purity as examined by 1H-NMR analysis (1H-NMR spectra of g-scale synthesis in ESI†), which can be used directly in subsequent steps for the synthesis of ipratropium bromide medicine.6
Flow synthesis is considered to be an interesting alternative to batch cell synthesis and electrochemical flow cells have been employed successfully for gram-scale synthesis.48 In order to make a comparison between the batch-cell and flow-cell synthesis of nortropanes in gram-scale, we used a newly designed electrochemical flow-cell,39 which has been applied to various electrochemical reactions.49,50 It is an 8-channel flow-cell, with a reactor volume of 88 μL per channel using graphite working and counter electrodes. The 8 channels of the flow-cell were used in series with the same solvent system as Table 5, entry 2. We performed a 2-gram scale electrosynthesis of noratropine. An isolated yield of 67% was obtained after three-step LLE, which is comparable to the batch-cell synthesis. A residence time of 42 min and 8 mA was found to be the optimal conditions. Increasing the applied current for the same residence time leads to lower yield and higher conversion to byproducts while decreasing the residence time at the same applied current did not lead to full conversion of atropine. It is noteworthy to mention that the ratio of the electrode surface area to the reactor volume of the batch-cell in gram-scale is calculated to be about 42 cm−1.51,52 This number is comparable to the surface-to-volume ratio of the flow-cell39 which is about 72 cm−1.
Since a considerable amount (80–90%) of non-aqueous waste generated from the manufacturing of active pharmaceutical ingredients (APIs) are solvents, the use of green solvents, such as ethanol, methanol and water, is a critical requirement.20 Besides, the application of heterogeneous catalysis plays a key role in developing new reaction processes to meet the target of green chemistry.53 The concept of catalyst immobilization and reuse, which has been recently defined as a new “key green chemistry research area” by big pharmaceutical companies,54 is addressed by our newly developed method using a cheap glassy carbon electrode for a prolonged period of time for different batches of the reaction in gram-scale. Finally, the purification of the synthesized noratropanes using a convenient LLE technique in high yields and purity is another advantage of the developed method avoiding a chromatographic purification step which is a growing demand in the pharmaceutical industry.45
A basic understanding of the mechanistic pathways of the catalytic reactions facilitates the improvement of catalytic processes providing the opportunity for the rational design of the catalytic systems.53 The high dissociation energy of the C–N bond and the intrinsic stability of amines makes cleavage of the C–N bond challenging for synthetic chemists.55 Generally, two types of C–N bond cleavage by transition-metal catalysts (such as iron-based catalysts) have been studied broadly: (a) the oxidative addition of transition-metal catalysts to the C–N bond and, (2) the formation of intermediate imine or iminium species.56 The latter mechanism has also been proposed for the Shono-type electrochemical oxidation,57,58 which provides a route to activate and functionalize C–H bonds in the vicinity of a nitrogen atom. Shono-type oxidations proceed by the initial formation of a nitrogen-centered radical via direct electron transfer to the electrode, followed by a sequence of ET/PT/ET (electron/proton/electron transfer) steps leading to an iminium intermediate, which can be trapped with water or alcoholic solvents.28,42,43 Therefore, we hypothesized that the electrochemical N-demethylation of tropane alkaloids on glassy carbon electrodes may also proceed via the formation of an iminium intermediate, which subsequently reacts with water to form nortropanes, as depicted in Fig. 2 (route a).
C–N bond cleavage in the form of N-dealkylation, a common in vivo metabolic pathway for most of the amine-containing drugs catalyzed by Cytochrome P450 (CYP) enzymes, also proceeds via iminium intermediate formation59,60 which can be trapped by a nucleophile such as cyanide.61 In order to investigate the mechanism, we added an excess amount of potassium cyanide as a source of cyanide ions to trap the iminium intermediate (Fig. 2, route c). LC–MS analysis of the outlet of the cell (Fig. S8†) showed that, besides intact atropine at m/z = 290.1 (M) and noratropine at m/z = 276.1 (M − 14), a new compound at m/z = 315.1 (M + 25) was produced during the reaction. This compound (N-nitrilo-noratropine, VI-1, Fig. 2) can be formed by the addition of a CN− group (26 Da) to an iminium intermediate and abstraction of a hydrogen atom, supporting a reaction mechanism during which an iminium intermediate is generated by a two-electron oxidation reaction at the anode. N-Nitrilo-noratropine was synthesized in a home-made electrochemical cell and characterized by 1H and 13C NMR. Dimer formation during the electrochemical reaction can be explained by this mechanism as well, since the produced noratropine, a secondary amine, can act as a nucleophile and react with the iminium intermediate resulting in a tertiary diamine (compound IV, Fig. 2) which undergoes a two electron electrochemical oxidation reaction to form an amidinium structure. The produced formaldehyde from the removed methyl group was detected by LC–MS using a simple derivatization method, where acetylacetone reacts with formaldehyde in the presence of ammonium acetate to form the cyclized 3,5-diacetyl-1,4-dihydrolutidine product (m/z of 194.1),62,63 which can be easily detected by LC–MS analysis (Fig. S12†). In theory, only 2 F mol−1 electricity is required for a two-electron oxidation reaction.47 The electrochemical N-demethylation of atropine at gram scale was calculated to require 2.6 F mol−1.
Footnote |
† Electronic supplementary information (ESI) available. See DOI: 10.1039/d0gc00851f |
This journal is © The Royal Society of Chemistry 2020 |