Series of methyl 3- and 5-(N-Boc-piperidinyl)-1H-pyrazole-4-carboxylates were developed and regioselectively synthesized as novel heterocyclic amino acids in their N-Boc protected ester form for achiral and chiral building blocks. In the first stage of the synthesis, piperidine-4-carboxylic and (R)- and (S)-piperidine-3-carboxylic acids were converted to the corresponding β-keto esters, which were then treated with N,N-dimethylformamide dimethyl acetal. The subsequent reaction of β-enamine diketones with various N-mono-substituted hydrazines afforded the target 5-(N-Boc-piperidinyl)-1H-pyrazole-4-carboxylates as major products, and tautomeric NH-pyrazoles prepared from hydrazine hydrate were further N-alkylated with alkyl halides to give 3-(N-Boc-piperidinyl)-1H-pyrazole-4-carboxylates. The structures of the novel heterocyclic compounds were confirmed by ¹H-, ¹³C-, and ¹⁵N-NMR spectroscopy and HRMS investigation.

A simple and efficient synthetic route to the novel 3a,4-dihydro-3H,7H- and 4H,7H-pyrazolo[4′,3′:5,6]pyrano[4,3-c][1,2]oxazole ring systems from 3-(prop-2-en-1-yloxy)- or 3-(prop-2-yn-1-yloxy)-1H-pyrazole-4-carbaldehyde oximes has been developed by employing the intramolecular nitrile oxide cycloaddition (INOC) reaction as the key step. The configuration of intermediate aldoximes was unambiguously determined using NOESY experimental data and comparison of the magnitudes of ¹J_CH coupling constants of the iminyl moiety, which were greater by approximately 13 Hz for the predominant syn isomer. The structures of the obtained heterocyclic products were confirmed by detailed ¹H, ¹³C and ¹⁵N NMR spectroscopic experiments and HRMS measurements.

A general approach towards the synthesis of tetrahydro-4H-pyrazolo[1,5-a][1,4]diazepin-4-one, tetrahydro[1,4]diazepino[1,2-a]indol-1-one and tetrahydro-1H-benzo[4,5]imidazo[1,2-a][1,4]diazepin-1-one derivatives was introduced. A regioselective strategy was developed for synthesizing ethyl 1-(oxiran-2-ylmethyl)-1H-pyrazole-5-carboxylates from easily accessible 3(5)-aryl- or methyl-1H-pyrazole-5(3)-carboxylates. Obtained intermediates were further treated with amines resulting in oxirane ring-opening and direct cyclisation—yielding target pyrazolo[1,5-a][1,4]diazepin-4-ones. A straightforward two-step synthetic approach was applied to expand the current study and successfully functionalize ethyl 1H-indole- and ethyl 1H-benzo[d]imidazole-2-carboxylates. The structures of fused heterocyclic compounds were confirmed by ¹H, ¹³C, and ¹⁵N-NMR spectroscopy and HRMS investigation.

In this study, we developed two complementary synthetic routes to novel piperidine- and azetidine-containing 2,3-disubstituted propanoic acid derivatives as heterocyclic amino acid building blocks. The strategy employs ketone- and carboxylic acid-derived Meldrum’s acid intermediates, which are converted into common α,β-unsaturated methyl esters through methanolysis; they are subsequently diversified via DBU-promoted aza-Michael addition with saturated cyclic amines and aromatic NH-heterocycles. The ketone-derived approach provided piperidine-containing derivatives in 35–89% yield and the corresponding azetidine analogues in 61–92% yield, whereas the complementary acid-derived route afforded regioisomeric products in 59–85% and 61–89% yield, respectively. Both synthetic sequences tolerated a broad range of nitrogen nucleophiles, and no alternative regioisomeric aza-Michael products were detected for heterocycles containing multiple nitrogen atoms. The structures of the synthesized compounds were established via ¹H, ¹³C, ¹⁵N, and ¹⁹F NMR spectroscopy together with HRMS, including detailed multidimensional NMR analysis of representative products. The developed methodology provides efficient access to structurally diverse heterocyclic propanoic acid derivatives and expands the repertoire of amino acid building blocks available for peptide chemistry, medicinal chemistry, and DNA-encoded library synthesis.

Heterocyclic unnatural amino acids and their biheterocyclic derivatives represent invaluable structural scaffolds in modern medicinal chemistry and peptidomimetics due to their ability to induce conformational constraints and modulate pharmacokinetic profiles. While saturated nitrogen heterocycles and monocycle heteroaromatic systems are well-established pharmacophores, research on linear biheterocyclic amino acid frameworks remains significantly underrepresented in the literature. Addressing this structural gap, this study aims to synthesize and characterize a novel series of functionalized pyrimidine (hetero)cyclic molecular hybrids acting as chiral heterocyclic amino acid derivatives. The target pyrimidine-5-carboxylates and pyrimidine-4-carboxylic acid derivatives were prepared from β-dicarbonyl compounds and their corresponding enamine analogues via cyclocondensation approaches. Particular attention was paid to reaction optimization, substrate scope exploration, and stereochemical integrity preservation, utilizing chiral HPLC analysis to evaluate enantiomeric retention. Pyrimidine-5-carboxylates prepared from β-enamino keto esters retained high enantiomeric excess (90.2–100% ee), whereas cyclization of β-diketones under strongly basic conditions at elevated temperature resulted in complete racemization (ee < 1%). Modification of the synthetic route and application of milder cyclization conditions partially suppressed racemization, affording pyrimidine derivatives with 61.8–65.5% ee. These findings suggest that substrate structure influences stereochemical integrity during pyrimidine synthesis.

An efficient and stereoselective method for the synthesis of chiral biheterocyclic N-Boc-protected pyrrolidine derivatives bearing indole, pyrazole, and indazole moieties is presented. In this approach, nucleophilic substitution of heterocyclic carboxylates with enantiomerically pure N-Boc-3-methanesulfonyloxypyrrolidines is used to afford N-(pyrrolidin-3-yl) derivatives in high yields with an inverted configuration. The methodology accommodates a range of substrates, enabling structural diversity. Reactions with pyrazole- and indazolecarboxylates generate regioisomeric products. Representative peptide-coupling reactions further demonstrated the synthetic utility of the synthesized chiral biheterocyclic pyrrolidine derivatives containing protected amino and carboxyl functionalities as building blocks for peptide synthesis. Halogenated indole and pyrazole derivatives underwent further functionalization via palladium-catalyzed cross-coupling to introduce aryl, heteroaryl, and alkynyl substituents. All of the N-Boc-substituted biheterocycle–pyrrolidine carboxylates displayed NMR spectra with two sets of signals, notable signal broadening, or both. These effects are due to the dynamic equilibrium between two conformers in a deuterated solvent and were studied in depth. The structures and stereochemistry of the synthesized compounds were confirmed by means of chiral HPLC, single-crystal X-ray diffraction, and advanced NMR analyses. This synthetic strategy provides access to chiral heterocyclic amino acid-like building blocks for peptide synthesis and medicinal chemistry.