www.nature.com/scientificreports Corrected: Publisher Correction OPEN Green Synthesis of Substituted Anilines and Quinazolines from Isatoic Anhydride-8-amide Received: 11 September 2019 Sudershan R. Gondi, Asim K. Bera & Kenneth D. Westover Accepted: 18 September 2019 Synthetic methods used to generate substituted anilines and quinazolines, both privileged Published: 03 October 2019 pharmacological structures, are cumbersome, hazardous or, in some cases, unavailable. We developed a straightforward method for making isatoic anhydride-8-amide from isatin-7-carboxylic acid as a tool to easily produce a range of quinazoline and substituted aniline derivatives using adaptable pH-sensitive cyclization chemistry. The approaches are inexpensive, simple, fast, efcient at room temperature and scalable, enabling the synthesis of both established and new quinazolines and also highly substituted anilines including cyano derivatives. Te 1,3-diazanaphthalene, or quinazoline, structural motif is a privileged pharmacological scafold1 found in a wide range of bioactive compounds designed for treating health conditions including cancer, infammation, hypertension, obesity, and infection2–6. Similarly substituted anilines are key elements of bioactive compounds7 and valuable industrial reagents8,9 (Fig. S1). Current synthetic methods involve using costly or hazardous reagents under harsh conditions to functionalize quinazolines and substituted anilines at late stages in synthetic schemes, and also require labor-intensive purifcation of chemical intermediates10,11. Moreover, options for functionalizing these structures are limited and challenging12. Easy Synthesis of IAA We developed a straightforward, broadly applicable, environmentally conscious method to prepare quinazoline and anilines analogues via isatoic anhydride-8-amide (IAA) afer observing that IAA can be prepared directly using the Schmidt reaction from 2,3-dioxoindoline-7-carboxylic acid 1a, also known as isatin-7-carboxylic acid, which can be obtained from anthranilic acid13. Te reaction occurred efciently at room temperatures in sulfuric acid and sodium azide in yields of ~85% (Fig. 1a). Under these conditions, IAA 2a precipitates, eliminating the need for additional purifcation or workup. Tis result surprised us because previous reports showed that treat- ment of isatin-7-carboxylic acid with sulfuric acid produces multiple products and, therefore, is unsuitable for synthetic workfows14. Nevertheless, we confrmed the identity of IAA as a single product by mass spectrometry, NMR, and x-ray crystallography (Fig. S2 and Table S1). We found that the reaction also occurred in yields of 50–75% with other azide sources such as tetrabutylammonium azide and tosyl azide, wherein in situ production of hydrazoic acid improves the safety of the reaction. However, sources such as acyl azide, alkyl azide, and alkoxy acyl azide were less efcient or did not produce IAA (Table 1). To our knowledge this is the frst example of using tosyl azide to yield benzoxazine. Alternative acids such as hydrochloric acid yielded no product, suggesting that sulfuric acid is critical to the mechanism. Reactions in DMSO produced isatin 7 acid-2-azide 2a2, suggesting that azide ions attack the C2-amide rather than the C3 carbonyl group without sulfuric acid (Table 1, entry 7). We found that IAA is an important intermediate for synthesizing a number of substituted anilines and quinazolines (Fig. 1b). Mechanism of IAA Formation We thought that IAA may form through a mechanism other than a typical Schmidt reaction for a ketone or free acid, wherein alkyl migration is accompanied by loss of carboxylic acid yielding an amine15. Instead, the mechanism occurred by solvolysis between the 2, 3 position, allowing direct azidation of the C3 keto of isatin-7-carboxylic acid and leading to a rightward (clockwise) cyclization between the NH-carbonyl and 7-acid to give isatoic anhydride (Fig. S3a). To confrm this mechanism, we evaluated isatin analogues with substitutions Departments of Biochemistry and Radiation Oncology, The University of Texas Southwestern Medical Center at Dallas, Dallas, Texas, 75390, United States. Correspondence and requests for materials should be addressed to K.D.W. (email: [email protected]) SCIENTIFIC REPORTS | (2019) 9:14258 | https://doi.org/10.1038/s41598-019-50776-y 1 www.nature.com/scientificreports/ www.nature.com/scientificreports Figure 1. (a) Synthesis of IAA. (b) Roadmap for synthesis of substituted anilines and quinazolines from IAA. at position 4, which we expected to compete for interactions with the amide intermediate in a counterclockwise (lefward) cyclization reaction, leading to the formation of phthalimide compounds (Fig. S3b). Consistent with this model, isatin-4-acid 1b, methyl ester 1c, primary amide 1d, C3-hydrazide 1e, and alkyl substitution at posi- tion 1 1f, 1g produced 3-amino-phthalimide 2b or 3-methylamino-phthalimide 2f (Table 2a, entries 1–6). As expected, moving the acid to the ffh position 1h prevents cyclization (Table 2a, entry 7). In addition to support- ing the proposed IAA mechanism, we also showed for the frst time how phthalimides, used in the synthesis of quinazoline 5-acid derivatives from an isatin derivative, are formed16. Tese sulfuric acid-based methods are eas- ier to perform than established procedures for generating phthalimides involving urea17, ammonium carbonate at elevated temperature, or reduction using palladium18. Tis method also provides synthetic pathways to a wide range of N-substituted phthalimides, whereas prior methods are limited to 3-methylamino-phthalimide17,19. While it is theoretically possible that the clockwise (rightward) cyclization required to form IAA may occur with either an acid, ester, or amide, the acid at position 7 is preferred (Table 2b, entries 8–9). Cyclization also depends on the availability of the secondary amine at position 1, as shown by failure to produce N-alkyl IAA when an alkyl group is introduced at position 1 (Table 2b, entry 10, 11). Surprisingly, alkyl substitution at frst position yielded a cyano aniline for both acid and esters 2k, 2l (Table 2b, entries 10, 11), suggesting that the com- bined efects of electron withdrawal by the acid and N-alkyl groups contribute to forming nitrile from the primary amide. Of note, known methods for the synthesis of n-alkylated nitrile-acid nitrile-ester involve using corrosive chemicals, high temperatures, and purifcation composed of multistep reactions, which are eliminated with the current method20. To assess whether incorporating a secondary amide at position 8 is possible, we started with enamines at position 3 1m, 1n, although they yielded IAA instead of a secondary amide (Table 2b, entries 12, 13). Tis fnding suggested that enamines are unstable under acidic conditions. Substitution on the phenyl ring with bromine at position 5 1o was also tolerated, giving 2o in excellent yield (Table 2b, entry 14). To exclude the pos- sibility that cyclization may occur using an n-alkyl acid at position 1 we tested reactions with isatin-N-propanoic acid and obtained an open chain product 2p (Table 2b, entry 15), instead of oxazine, suggesting that oxazine is less stable than benzoxazine. Our synthetic method simplifes isatin transformations considerably compared to prior methods. Synthesis of isatoic anhydride from isatin previously required using reagents such as peroxides21,22, phenyliodide23, and NBS24; making benzamide derivatives from isatin required using chromic acid25, peroxide/phosphate bufer systems26, or SCIENTIFIC REPORTS | (2019) 9:14258 | https://doi.org/10.1038/s41598-019-50776-y 2 www.nature.com/scientificreports/ www.nature.com/scientificreports Entry Azide source Solvent Yield (%) 1 Sodium Azide H2SO4 85 2 Tosyl Azide H2SO4 75 3 Tetrabutyl Azide H2SO4 50 4 RCON3 H2SO4 5 5 ROCON3 H2SO4 0 6 RN3 H2SO4 0 7 Sodium Azide DMSO 0 8 Sodium Azide HCl 0 Table 1. IAA reaction optimization. in situ generated hydrazoic acid27,14; and generating amino benzamide derivatives from isatoic anhydride required using ammonia22,28 and ammonium carbonate29. In contrast, we can now approach multiple classes of derivatives from IAA using mild, established reactions. Scope of Substituted Anilines Originating from IAA We recognized that IAA could be used as a starting material for producing a range of useful substituted anilines and quinazoline derivatives in situ, primarily by manipulating pH, with either neutral or acidic conditions giving substituted anilines where position 1 (Table 3, R1) can include a range of substituents including acids, esters, thioesters, amides, and halogens. Conversion of IAA to the open chain 3-acid-2-amino-benzamide 3a is read- ily accomplished at pH 7 and also by heating in sulfuric acid (Table 3, entry 1). Next, the corresponding ester, 3-ethylester, 2-amino-1-bezamide 4a was prepared by allowing IAA to react with polar solvents (ethanol) under refux conditions (Table 3, entry 2). Alternatively, esters can be achieved by allowing IAA to react with K2CO3 in a polar solvent at room temperature. We showed that multiple ester derivatives 4a, 4b including a thioester 5 could be obtained using this method (Table 3, entry 3). Because transformation of isatoic anhydride to 2-amino-benzamide using ammonium carbonate in dioxane was previously reported29, IAA was treated under these conditions giving 2-amino, 1,3 benzadiamde, in quantita- tive yield 6 (Table 3, entry 4). Te same product can also be obtained by treating IAA with ammonium carbonate or