Shuang-Qi Tang

Maeva Leloire

Severine Schneider

Julie Mohr

Jacques Bricard

Patrick Gizzi

Delphine Garnier

Martine Schmitt

Frédéric Bihel
email: fbihel@unistra.fr

Séverine Schneider

Diastereoselective Synthesis of Nonplanar 3-Amino-1,2,4-oxadiazine Scaffold: Structure Revision of Alchornedine

de niveau recherche, publiés ou non, émanant des établissements d'enseignement et de recherche français ou étrangers, des laboratoires publics ou privés.

INTRODUCTION

Limitations in solubility, pharmacokinetics, and bioavailability of highly aromatic molecules have been well recognized; 1 the medicinal chemistry community has become increasingly aware of the requirement to create drug-like compounds containing saturated building blocks. [2][START_REF] Kumar | Synthesis of a Family of Spirocyclic Scaffolds: Building Blocks for the Exploration of Chemical Space[END_REF][4] Among them, the six-membered heterocycle oxadiazines containing one oxygen and two nitrogen atoms have been considered as important skeletons because of their interesting potential in organic and medicinal chemistry. [START_REF] Zhang | Discovery of a Novel Series of Tricyclic Oxadiazine 4a-Methyl Esters Based on Indoxacarb as Potential Sodium Channel Blocker/Modulator Insecticides[END_REF][START_REF] Eissa | Green Synthesis, Antibacterial, and Antifungal Activities of 1,3,4-Oxadiazines[END_REF][7][START_REF] Lee | Discovery of highly selective and potent monoamine oxidase B inhibitors: Contribution of additional phenyl rings introduced into 2aryl-1,3,4-oxadiazin-5(6H)-one[END_REF][START_REF] Ke | Synthesis and Biological Properties of Dihydro-Oxadiazine-Based Heterocyclic Derivatives[END_REF][START_REF] Abele | Oximes of six-membered heterocyclic compounds with two and three heteroatoms. II.* Reactions and biological activity[END_REF] Unlike their aromatic triazine counterparts, the bivalent character of the oxygen atom endows oxadiazines with an interesting nonaromatic and non-planar geometry which has recently been highlighted and utilized in drug discovery through the concept of "escape from flatland" developed by F. Lovering et al. [START_REF] Lovering | Escape from flatland: increasing saturation as an approach to improving clinical success[END_REF][START_REF] Lovering | Escape from Flatland 2: complexity and promiscuity[END_REF] Among the different types of oxadiazines classified on the basis of the positions of their heteroatoms, we focused our work on the 3-amino-1,2,4-oxadiazine (AOXD) scaffold (Figure 1). This scaffold is particularly interesting because it presents two stereogenic sp 3 centers; the presence of the bivalent oxygen atom in the α-position of the guanidine moiety affects pKa, strongly decreasing the basicity of the guanidine moiety. This alkoxy-guanidine moiety is well known in its acyclic form in the natural amino acid L-canavanine, which possesses arginine antimetabolite property and is synthesized by leguminous plants. [START_REF] Thomas | Toxicity and pharmacokinetics of the nonprotein amino acid l-canavanine in the rat[END_REF] However, to date, only one natural product has been reported that consists of the cyclic form of AOXD. This natural guanidine-based alkaloid, alchornedine (Figure 1), was isolated from the leaves of Alchornea glandulosa and structurally identified by Barrosa et al in 2014. [START_REF] Barrosa | Alchornedine, a new anti-trypanosomal guanidine alkaloid from Alchornea glandulosa[END_REF] The authors also showed that alchornedine displayed antiprotozoal activity against Trypanosoma cruzi (Y strain). However, the total synthesis of this compound has not been reported yet, limiting the optimization of its activity against American trypanosomiasis.

Because of its distinctive structure and properties, general synthetic methods for chiral AOXD using readily available reagents are desirable. Although several preparative methods have been described for 3-alkyl/aryl-1,2,4-oxadiazines, [15][START_REF] Shetnev | The latest advances in chemistry of 1,2,4-oxadiazines[END_REF][START_REF] Pathipati | Lewis Acid Catalyzed Annulation of Nitrones with Oxiranes, Aziridines, and Thiiranes[END_REF] the synthesis of chiral AOXD, to our knowledge, has not been reported. In this study, we established a general synthetic method to access chiral AOXD by using readily available N-Boc-amino acids as starting materials. The first objective of this study was the synthesis of racemic AOXD and its geometry has been confirmed by X-ray crystallography. Then, several conditions were screened for the diastereoselective synthesis of chiral AOXDs and the relationship between chirality and NMR characteristics was highlighted. As a novel scaffold, the physicochemical properties of AOXD were also investigated. Finally, alchornedine was synthesized, and its spectra and analytical data were compared with those of the isolated compound previously reported. [START_REF] Barrosa | Alchornedine, a new anti-trypanosomal guanidine alkaloid from Alchornea glandulosa[END_REF]

RESULTS AND DISCUSSION

As an initial experiment, achiral N-Boc-glycine was used as the starting material for exploring the synthesis of racemic AOXD (Scheme 1). N-Boc-glycine reacted with N, N'-carbonyl diimidazole (CDI) and N,O-dimethylhydroxylamine to form the corresponding Weinreb amide (2) in 99% yield. [START_REF] Oukoloff | Design, synthesis and evaluation of photoactivatable derivatives of microtubule (MT)-active [1,2,4]triazolo[1,5-a]pyrimidines[END_REF] The nucleophilic addition between 2 and 3,4-dichloro-phenyl magnesium bromide furnished ketone 3 in 86% yield, which was directly reduced by NaBH4, to yield alcohol 4 in 99% yield. As a key step in AOXD synthesis, the Mitsunobu reaction between alcohol 4 and N-hydroxyphthalimide (NHPI) was conducted to form the C-ON bond, affording the key intermediate, 5, in 94% yield. Then, phthaloyl and Boc protections were successively removed by hydrazine hydrate and HCl treatments, affording 6 and 7 in 94% and 96% yield, respectively. Finally, the cyclization reaction of 7 was carried out with BrCN, to afford racemic AOXD 8 and its 4-cyanated side product 9 in 81% and 17% yield, respectively (see SI section I). The presence of such a side product indicated that the nitrogen atom at position 4 (-HN-CH2-) in 8 is more nucleophilic toward BrCN than those at positions 3 and 2, respectively. X-ray crystallography confirmed the AOXD structure of 8 (Scheme 1), with a C=N double bond located between N 2 and C 3 , rather than C [START_REF] Kumar | Synthesis of a Family of Spirocyclic Scaffolds: Building Blocks for the Exploration of Chemical Space[END_REF] and N 4 , likely because of the stabilization of p-π conjugation between the oxygen atom and the C=N double bond. Meanwhile, the bivalent character of the oxygen atom induced the expected non-planar geometry of AOXD, in which C [START_REF] Eissa | Green Synthesis, Antibacterial, and Antifungal Activities of 1,3,4-Oxadiazines[END_REF] is located far out of the plane constituted by the N 2 =C 3 -N 4 moiety. Moreover, the N 2 -O 1 and N 4 -C [START_REF] Zhang | Discovery of a Novel Series of Tricyclic Oxadiazine 4a-Methyl Esters Based on Indoxacarb as Potential Sodium Channel Blocker/Modulator Insecticides[END_REF] bonds are unparallel because of the different bond angles between N 2 -O 1 -C [START_REF] Eissa | Green Synthesis, Antibacterial, and Antifungal Activities of 1,3,4-Oxadiazines[END_REF] and N 4 -C 5 -C [START_REF] Eissa | Green Synthesis, Antibacterial, and Antifungal Activities of 1,3,4-Oxadiazines[END_REF] . Despite the absence of any functional groups at C 5 of AOXD, or on ortho position of the phenyl ring, the oxadiazine ring in 8 is perpendicular to the phenyl ring, which allows the hydrogen atom C [START_REF] Eissa | Green Synthesis, Antibacterial, and Antifungal Activities of 1,3,4-Oxadiazines[END_REF] H to be in the same axis as the aromatic ring (for more details of crystallographic data and structure refinement parameters, see SI). After determining the synthesis method of racemic AOXD, we selected N-Boc-L-alanine, N-Boc-L-valine and N-Boc-L-proline as starting materials for the synthesis of chiral AOXDs exhibiting two stereogenic centers (Table 1). Using the synthetic approach described in Scheme 1, the N-Boc-L-amino acids were transformed to their corresponding ketones through the reactions of their corresponding Weinreb amides with PhMgBr, affording 10, 11 and 12 in 86%, 83% and 83% overall yield, respectively. To reduce ketones 10-12 to their corresponding alcohols, several reductants were evaluated to determine optimal stereoselective conditions allowing one diastereomer to be favored over another (Table 1). Several reduction methods of α-amino ketones have been published recently; [START_REF] Kasamatsu | α-Arylation of α-Amino Acid Derivatives with Arynes via Memory of Chirality: Asymmetric Synthesis of Benzocyclobutenones with Tetrasubstituted Carbon[END_REF][START_REF] Ripa | Discovery of a Novel Oral Glucocorticoid Receptor Modulator (AZD9567) with Improved Side Effect Profile[END_REF][START_REF] Giltrap | Synthetic Studies Toward the Skyllamycins: Total Synthesis and Generation of Simplified Analogues[END_REF][START_REF] Trost | Direct catalytic enantioselective amination of ketones for the formation of tri-and tetrasubstituted stereocenters[END_REF][START_REF] Ohsawa | Total Synthesis and Structural Revision of Cyclotetrapeptide Asperterrestide A[END_REF] however, none of them showed a systematic comparison of the reduction of α-aminoketones by borane-or aluminium-based reducing agents exhibiting more or less bulkiness. [a] Entry Reducing agents Isolated yield (%) [b]

Ala

Val Pro 13a 13b 14a 14b 15a 15b 3 [d] (S)-Me-CBS, BH3 97 0 99 0 70 29 4 [d] (R)-Me-CBS, BH3 [START_REF] Guo | Synthesis of Chiral Aminophosphines from Chiral Aminoalcohols via Cyclic Sulfamidates[END_REF][START_REF] Lee | Stereoselective Synthesis of Norephedrine and Norpseudoephedrine by Using Asymmetric Transfer Hydrogenation Accompanied by Dynamic Kinetic Resolution[END_REF][START_REF] Schmidt | Stereocontrolled Synthesis of β-Amino Alcohols from Lithiated Aziridines and Boronic Esters[END_REF][START_REF] Bilke | Catalytic Asymmetric Synthesis of Piperidines from Pyrrolidine: Concise Synthesis of L-733,060[END_REF] [c] THF was replaced with MeOH (1 mL). [d] (S)-or (R)-(-)-2-methyl-CBSoxazaborolidine (0.01 mmol), BH3-Me2S complex (0.2 mmol) were used.

Partial diastereoselectivity was observed when NaBH4 was used (Table 1, entry 1), privileging formation of the (S,R) diastereomer for Ala-and Val-type ketones (77% (S,R) 13a and 22% (S,S) 13b; 82% (S,R) 14a and 18% (S,S) 14b, respectively), while the (S,S) diastereomer was favored for the Pro-type ketone (24% (S,R) 14a and 72% (S,S) 14b). The bulky organoborane L-selectride led to a weak diastereoselective reduction of ketones 10-12 (Table 1, entry 2). As a well-known chiral reaction, Corey-Bakshi-Shibata (CBS) reduction was also attempted. [START_REF] Helal | The Corey-Bakshi-Shibata Reduction: Mechanistic and Synthetic Considerations -Bifunctional Lewis Base Catalysis with Dual Activation[END_REF] (S)-2-Me-CBS yielded only the (S,R) diastereomer in nearly quantitative yield for Ala and Val-type ketones (97% 13a; 99% 14a) (Table 1, entry 3), but appeared less efficient for the Pro-type ketone, leading to diastereomers (S,R) 15a and (S,S) 15b in 70% and 29% yield, respectively (Table 1, entry 3). In contrast, (R)-2-Me-CBS led to low diastereoselectivity regardless of the bulkiness of the ketone (Table 1, entry 4). Further, Lithium aluminium hydride (LiAlH4) exhibited low diastereoselectivity toward bulky ketones 11 and 12, favoring the formation of (S,R) 13a (74% yield) starting from Ala-type ketone 10 (Table 1, entry 5). In contrast to the bulky organoborane Lselectride, the bulky aluminum hydride DIBAL-H diastereoselectively reduced ketones 10-12, yielding only the single (S,R) diastereomers 13a-15a in nearly quantitative yield (Table 1, entry 6). This excellent diastereoselectivity of DIBAL-H could be explained by the good complexation between the aluminum atom and the carbonyl oxygen atom as well as the steric hindrance between the isobutyl moieties of DIBAL-H and the Boc group following the Felkin-Anh model (see SI section II).

Next, the Mitsunobu reaction with NHPI was conducted to introduce the C-ON bond on the chiral alcohols 13-15 (Figure 2A-D). As expected, key intermediates 16 and 17 were obtained with the inversion of configuration in 74%-96% yield from the corresponding Ala-and Val-type alcohols (Figures 2A and2B). However, with Pro-type (S,R) alcohol 15a, the Mitsunobu reaction led to the unexpected retention of configuration, affording a key intermediate 18a (S,R) in 94% yield (Figure 2C).

Intermediate 18a was first characterized by NMR, and the retention of configuration was confirmed by the X-ray crystallography of AOXD 21, which was obtained from the cyclization of deprotected 18a with BrCN (Figure 2E). Although it is not a frequent case, such configuration retention has been reported wherein the Mitsunobu reaction was performed on sterically hindered alcohols (see SI section III). [START_REF] Swamy | Mitsunobu and Related Reactions: Advances and Applications[END_REF] Different reactivities were also observed between (S,R) and (S,S) diastereomers 13-15. Moreover, (S,R) alcohols 13a-15a underwent the Mitsunobu reaction at 0 °C (Figures 2A and2C), while a higher temperature (rt or 45 °C) and an additional equivalent of reagents were required for the reactions involving (S,S) Ala-and Val-type alcohols 13b and 14b (Figure 2B). The Mitsunobu reaction of the (S,S) Pro-type alcohol 15b yielded a complex mixture at 45 °C, possibly because of the hindrance of the Boc group under the configuration-retained mechanism (Figure 2D, also see SI section IV).

Finally, after deprotection of the phthaloyl and Boc moieties, key intermediates 16-18 were cyclized in the presence of BrCN to obtain the corresponding diastereomeric AOXDs 19-21 after three steps in 56%-88% overall yield. (Figures 2A-C). Interestingly, cyanamide 22 could be isolated as an intermediate of the cyclization reaction of 18a with BrCN (Figure 2D, dotted box), indicating that the secondary amine of the pyrroline moiety is more nucleophilic than alkoxyamine toward BrCN (see SI section V). The X-ray crystallographic structures of 19a and 21 were obtained as representatives of (S,S) and (S,R) AOXDs (Figure 2E). To facilitate the identification of diastereomeric AOXDs, the 1 H chemical shifts (δ) and coupling constants (J) of 19-21 in DMSO-d [START_REF] Eissa | Green Synthesis, Antibacterial, and Antifungal Activities of 1,3,4-Oxadiazines[END_REF] were determined and significant differences were observed. In particular, the proton of the >CH'-Omoiety in (S,S) 19a and 20a was detected at 3.82-4.09 ppm, with a coupling constant (JH') of 8.2-8.5 Hz (Figure 2F). In contrast, the corresponding proton in (S,R) 19b, 20b and 21 was observed at 4.55-4.98 ppm, with JH' of about 3-4.3 Hz.

The AOXDs described herein have not been previously characterized in terms of physicochemical properties. The presence of an oxygen atom in the α-position of the guanidine moiety would likely affect pKa as well as the distribution coefficient log D. These parameters were experimentally measured and compared with those of the cyclic guanidine analogs 23-25 (Figure 3). Because of the strong basicity of guanidine, the pKa of 23 was too high to be measured under the titration condition and was estimated to be >13. A negative value of logD (D7.4 = -0.64 ± 0.06) indicates that 23 is hydrophilic and unfavorable for membrane permeability by passive diffusion. Furthermore, the introduction of one double bond in 23 resulted in the formation of unsaturated cyclic guanidine 24. As expected from the conjugation between the benzene ring and the the guanidine moiety, pKa decreased to 11.37 and log D7.4 increased to 1.90 ± 0.04. Such a strong increase in log D is because of the decrease in pKa, and the presence of both chlorine atoms on the aromatic ring. Next, the unsaturated carbon atom adjacent to the guanidine moiety in 24 was replaced by a nitrogen atom, leading to the formation of cyclic iminoguanidine 25. The resulting pKa dropped to 10.02 and log D7. 4 that can be attributed to the transposition of the α-nitrogen atom in 25 by an oxygen atom, dramatically decreasing pKa to 6.91 with a suitable log D7.4 at 1.88 ± 0.06. Diastereomers 20b and 21 were slightly more basic and showed pKa values of 7.14 and 7.44, respectively. Distribution coefficient log D7.4 of 8, 20b, and 21 was <2 despite the presence of the substituents. This is a promising phenomenon as it allows a comfortable margin of maneuver compared to the Lipinski recommendations. [START_REF] Benet | BDDCS, the Rule of 5 and drugability[END_REF] Compared to non-conjugated cyclic guanidine 23, AOXD 8, 20b and 21 are more than six orders of magnitude less basic. Interestingly, at the physiological pH 7.4, a significant proportion of AOXDs are not protonated, improving their ability to cross biological barriers, unlike compounds 23-25 that only exist in their protonated forms at physiological pH. Finally, we determined the metabolic stability of AOXDs 8, 20b, and 21 in the presence of mouse liver microsomes (MLM), and all of them exhibited complete stability after 1 h. For comparison, testorenone and dextrometorphan, FDA-approved drugs used as positive controls in this assay, showed half-lives of 5 and 19 min, respectively. These distinctive physicochemical properties and its stability in aqueous biological media make AOXD a promising scaffold for drug development. Furthermore, the proposed synthetic approach for AOXD was utilized toward the total synthesis of alchornedine, the only natural product exhibiting a 3-amino-1,2,4-oxadiazine (AOXD) scaffold. Starting from N-Boc-glycine, AOXD 31 was synthesized as described previously (Scheme 2), except for the reduction of α, β-unsaturated ketone 26, which was performed with NaBH4 in the presence of CeCl3. AOXD 31 was obtained from 2 after six steps and 61% overall yield. Direct alkylation of 31 using prenyl bromide in the presence of TEA afforded 32 in 74% yield (Scheme 2). To confirm the position of the prenyl moiety, we synthesized its 15 N-labeled analog (33) based on the principle that NMR signals of hydrogen or carbon atoms directly linked to 15 N will split. Following the synthetic method shown in Scheme 2, 33 was obtained after eight steps and 40% overall yield starting from 15 N-Boc-glycine (see SI section VI). The position of the prenyl moiety in 33 could be easily identified in the 1 H NMR spectra based on the presence of the -NH2 signal as well as the split of [START_REF] Thomas | Toxicity and pharmacokinetics of the nonprotein amino acid l-canavanine in the rat[END_REF] C signals of >C= 15 N-and = 15 N-CH2-, while no split was observed for >N-CH2-from the prenyl moiety (see SI section VII). To modify the reactivity of 31 towards electrophiles, the nitrogen atom at position 2 was protected in the presence of (Boc)2O, affording 34 in 91% yield (Scheme 2). The position of the Boc group was characterized unambiguously in 2D NMR (see SI section VIII). The amino group in 34 was then deprotonated by NaH and alkylated by prenyl bromide in THF, producing 35 in 76% yield. After Boc deprotection by HCl in dioxane, the structure of alchornedine 1, as proposed by Barrosa et al., [START_REF] Barrosa | Alchornedine, a new anti-trypanosomal guanidine alkaloid from Alchornea glandulosa[END_REF] was obtained in 96% yield. Full structural characterization of 1 is shown in SI section IX, including 1 H and [START_REF] Thomas | Toxicity and pharmacokinetics of the nonprotein amino acid l-canavanine in the rat[END_REF] C NMR, 2D-NMRs, and HRMS.

Scheme 2. Synthesis of of alchornedine 1.

To confirm the structure of alchornedine, NMR spectra of 1 were compared with those reported for alchornedine (Figure 4A, SI section X). [START_REF] Barrosa | Alchornedine, a new anti-trypanosomal guanidine alkaloid from Alchornea glandulosa[END_REF] Significant differences were found in both chemical shifts and coupling constants suggesting that Barrosa's proposed structure for alchornedine was incorrect. Such an error could be explained by the fact that all NMRs analyses by Barrosa et al. were performed in MeOD, which prevented the observation of all signals corresponding to NH, which are the only protons between the prenyl group and CH2 at position 5 of the 1,2,4-oxadiazine scaffold.

The reported mass spectrum of alchornedine shows a molecular peak at 210.16 (M+H), as well as a peak of fragmentation at 194.16, corresponding to the loss of a hydroxyl moiety. This result is not consistent with the 3-amino-1,2,4oxadiazine scaffold, but rather with that of the imidazolidine-1-ol scaffold shown in Figure 4B. Moreover, the NOESY signal reported for alchornedine between CH2 in the ring and CH2 of the prenyl moiety is attributed to the presence of the imidazolidine-1-ol scaffold. Furthermore, this imidazolidin-1-ol scaffold is identical to the reported structures of natural products, as it is found in alkaloids such as alchornein and isoalchornein. [START_REF] Büchi | Syntheses of (±)-Alchorneine and (±)-Isoalchorneine[END_REF] Finally, it can be noticed that both the alkaloids correspond to a cyclized form of the structure we propose for alchornedine. However, the synthesis of this structure (Fig 4B ) is not trivial and is under progress in our laboratory to confirm our hypothesis.

Conclusions

A general method for the synthesis of 3-amino-1,2,4oxadiazine (AOXD) scaffold was developed by using commercially available N-Boc amino acids as starting materials. We developed diastereoselective reactions to synthesize chiral compounds exhibiting non-planar geometries and promising physicochemical properties, because of which the compounds could be applicable in medicinal chemistry approaches. Based on the established method, alchornedine was synthesized and its structure was determined; however, spectral inconsistences between the synthesized alchornedine and the one isolated from the leaves of Alchornea glandulosa indicated that the proposed structure of alchornedine is incorrect. The total synthesis of alternative structures of alchornedine is currently underway. The determination of the correct structure of alchornedine will benefit medicinal chemistry, because alchornedine exhibits promising antiprotozoal activity against American trypanosomiasis. 14

EXPERIMENTAL SECTION General information and typical procedures

All commercial reagents were used without additional purification. Analytical TLC was performed using silica gel plates Merck 60 F254 and plates were visualized by exposure to ultraviolet light (254 nm). Compounds were purified on silica gel Merck 60 (particle size 0.040-0.063 nm). 1 H, [START_REF] Thomas | Toxicity and pharmacokinetics of the nonprotein amino acid l-canavanine in the rat[END_REF] C and 15 N NMR spectra were recorded on Bruker Avance Spectrometer operating at 400 MHz, 101 MHz and 51 MHz, respectively. Structural assignments were made with additional information from gCOSY, gHSQC, and gHMBC experiments. All chemical shift values δ and coupling constants J are quoted in ppm and in Hz, respectively, multiplicity (s= singulet, d= doublet, t= triplet, q= quartet, m = multiplet, br = broad). Analytical RP-HPLC-MS was performed using a LC 1200 Agilent with quadrupole-time-offlight (QTOF) (Agilent Accurate Mass QToF 6520) with a Zorbax Agilent C18-column (C18, 50 mm × 2.1 mm; 1.8 µm) using the following parameters: 1) The solvent system: A (0.05% of formic acid in acetonitrile) and B (0.05% of formic acid in H2O); 2) A linear gradient: t = 0 min, 98% B; t = 8 min, 0% B; t = 12.5 min, 0% B; t = 12.6 min, 98% B; t = 13 min, 98% B; 3) Flow rate of 0.5 mL/min; 4) Column temperature: 35 °C; 5) DAD scan from 190 nm to 700 nm; 6) Ionization mode : ESI + or ESI -. HPLC were performed using a Dionex UltiMate 3000 using the following parameters: Flow rate of 0.5 mL/min, column temperature: 30 °C, solvent system: A (MeOH) and B (0.05% of TFA in H2O), t = 0 min to 1 min: 50 to 60% of B, then t = 1 min to t = 10 min: 60 to 100% of B and t = 10 min to t = 15 min: 100% of B. The singlecrystal structure analysis was performed using X-ray diffraction with a Thermo Fisher ESCALAB 250 diffractometer.

Compound 23 -25 were prepared according to previous reported literatures. [27][START_REF] Baskaran | A facile reduction of 2-aminopyrimidines with triethylsilane and trifluoroacetic acid[END_REF][START_REF] Nguyen | Structure-Activity Relationship Study around Guanabenz Identifies Two Derivatives Retaining Antiprion Activity but Having Lost α2-Adrenergic Receptor Agonistic Activity[END_REF][START_REF] Hammoud | Aminoguanidine Hydrazone Derivatives as Nonpeptide NPFF1 Receptor Antagonists Reverse Opioid Induced Hyperalgesia[END_REF] 1. Typical procedure 1 (TP1) for the preparation of Weinreb amides 2, 36 -38 and 45 CDI (1.5 eq.) was added slowly to a solution of N-Boc-amino acid (1 eq.) in DCM and stirred at rt for 30 minutes until CO2 evolution ceased. Then, TEA (1.5 eq.) was added dropwise to the reaction mixture followed by the addition of N,Odimethylhydroxylamine hydrochloride (1.5 eq.). After the addition was complete, the resulting mixture was stirred at rt for an additional 16 h. The reaction was then quenched by addition of water and the resulting solution was extracted with DCM thrice. The combined organic phases were washed with brine, dried over anhydrous Na2SO4 and filtered. The filtrate was concentrated in vacuo without further purification to afford the desired Weinreb amide. 2. Typical procedure 2 (TP2) for the preparation of ketones 3, 10 -12, 26 and 46 Grignard reagent (1.5 ~ 3.5 eq.) in THF was added dropwise to a solution of Weinreb amide (1 eq.) in THF at 0 °C under Ar. After the addition was complete, the reaction mixture was stirred at 0 °C overnight. The reaction mixture was then poured slowly into saturated NH4Cl aqueous solution at 0 °C. After stirring for an additional 10 min, the resulting mixture was extracted with EtOAc thrice. The combined organic phases were washed with brine, dried over anhydrous Na2SO4 and filtered. The filtrate was concentrated in vacuo and purified by silica gel column chromatography to afford the desired ketone. 3. Typical procedure 3 (TP3) for the preparation of alcohols 4, 13 -15, 27 and 47 Method 1 associate with the use of NaBH4: NaBH4 (4 eq.) was added portionwise to a solution of ketone (1 eq.) in MeOH at 0°C and the resulting mixture was stirred at rt for an additional 6 h. The mixture was then poured slowly into a chilled saturated NH4Cl aqueous solution, stirred for an additional 10 min. The resulting mixture was concentrated in vacuo to remove MeOH and then extracted with EtOAc thrice. The combined organic phases were washed with brine, dried over anhydrous Na2SO4 and filtered. The filtrate was concentrated in vacuo and purified by silica gel column chromatography or reverse phase C18 column chromatography to afford the desired alcohol. Method 2 associate with the use of DIBAL-H: DIBAL-H (1 M in THF, 2 eq.) was added dropwise to a solution of ketone (1 eq.) in dry THF maintained at 0°C under Ar and the reaction was stirred at rt for 6 h. The reaction mixture was then poured slowly into a chilled saturated NH4Cl aqueous solution, stirred for an additional 10 min and then extracted with EtOAc thrice. The combined organic phases were washed with brine, dried over anhydrous Na2SO4 and filtered. The filtrate was concentrated in vacuo without further purification to afford the desired alcohol. Method 3 associate with the use of (R) -2-Me-CBS and borane dimethylsulfide complex: Borane dimethylsulfide complex (2 eq.) was added dropwise to a solution of (R)-2-Me-CBS-oxazaborolidine (0.4 M in THF, 0.1 eq.) in THF at 0 °C under Ar and the resulting solution was stirred for 20 min. The ketone solution (1 eq) in THF was added slowly at 0°C to the borane solution over a period of 15 min, the ice bath was removed, and the resulting mixture was stirred at rt for 16 h. The reaction solution was poured slowly into a chilled saturated citric acid aqueous solution (30 mL). After stirring for an additional 10 min, the resulting mixture was extracted with EtOAc thrice. The combined organic phases were concentrated in vacuo and purified by reverse phase C18 column chromatography to afford the desired alcohol. Method 4 associate with the use of LiAlH4: A solution of ketone (1 eq.) in dry THF was added dropwise to a suspension of LiAlH4 (2 eq.) in THF at 0°C under Ar and then stirred at rt for 6 h. The reaction mixture was poured slowly into a chilled saturated NH4Cl aqueous solution, and stirred for an additional 10 min while the temperature was kept at 0°C. The resulting mixture was then extracted with EtOAc thrice. The combined organic phases were washed with brine, dried over anhydrous Na2SO4 and filtered. The filtrate was concentrated in vacuo and purified by silica gel column chromatography to afford the desired alcohol. Method 5 associate with the use of NaBH4 and CeCl3: Trichlorocerium heptahydrate (1.6 eq.) was added to a solution of ketone (1 eq.) in MeOH and sonicated until the suspension became clear. Then, the mixture was cooled to -70 °C, NaBH4 (1.5 eq.) was added portionwise and the reaction mixture was stirred at -70 °C for an additional 1 h. The reaction mixture was allowed to warmed-up to -20 °C and then quenched carefully by the addition of chilled H2O. The crude was concentrated in vacuo to remove MeOH and the aqueous suspension was extracted with EtOAc thrice. The combined organic phases were washed with brine, dried over anhydrous Na2SO4 and filtered. The filtrate was concentrated in vacuo and purified by silica gel column chromatography to afford the desired alcohol. 4. Typical procedure 4 (TP4) for the preparation of the PINO derivatives 5, 16 -18, 28 and 48 The solution of corresponding alcohol 4, 13-15 or 27 (1 eq), NHPI (1.5 ~ 3 eq) and PPh3 (1.8 ~ 3.5 eq.) in dry THF was cooled 0 °C, degassed and backfilled with Ar thrice. Then, DEAD (1.8 ~ 3.5 eq.) was added dropwise at 0 °C under Ar and allowed to stir at the given temperature for 16 h. The reaction mixture was quenched by the addition of water and extracted with EtOAc thrice. The combined organic phases were washed with brine, dried over anhydrous Na2SO4 and filtered. The filtrate was concentrated in vacuo and purified by silica gel column chromatography to afford the desired product. 5. Typical procedure 5 (TP5) for the preparation of compounds 6, 29, 39, 41, 43 and 49 Hydrazine hydrate (15 eq.) was added dropwise to a solution of N-alkoxyl phthalimide (1 eq.) in EtOH and stirred at rt for 5 h. The reaction mixture was concentrated in vacuo and purified by silica gel column chromatography to afford the desired product. 6. Typical procedure 6 (TP6) for the preparation of compounds 7, 30, 40, 42, 44 and 50 Boc amino alkoxyamine (1 eq.) was added to a hydrogen chloride solution in dioxane (4M) and stirred at rt for 16 h. The reaction was concentrated in vacuo and dried under reduced pressure overnight to afford the alkoxyamines as a dihydrochloride salt. 7. Typical procedure 7 (TP7) for the preparation of AOXDs 8, 19 -21, 31and 51 TEA (4 eq.) was added dropwise to a solution of amino alkoxyamine dihydrochloride (1 eq.) in dry MeCN at 0 °C, under argon and the mixture was stirred for 5 min. Then a solution of BrCN (1.1 eq.) in MeCN was added dropwise and stirring was maintained for 30 min at 0°C. The ice bath was removed and the reaction stirred for 16 h while warming to ambient temperature. The crude was concentrated in vacuo and purified by reverse phase C18 column chromatography (MeCN/H2O) to afford the desired AOXD.

N-(3-methylbut-2-en-1-yl)-6-(prop

Tert-butyl (2-(methoxy(methyl)amino)-2-oxoethyl)carbamate (2).

Following general procedure TP1 and starting from N-Boc glycine (1 eq., 4 g, 22.8 mmol), CDI (1.5 eq., 5.55 g, 34.2 mmol), TEA (1.5 eq, 4.76 mL, 34.2 mmol) and N,Odimethylhydroxylamine hydrochloride (1.5 eq., 3.34 g, 34.2 mmol) in DCM (50 mL), the title compound 2 was obtained as a white solid (4.94 g, 99%). 1 H NMR (400 MHz, CDCl3): δ 5.26 (s, 1H), 4.07 (d, J = 3.8 Hz, 2H), 3.70 (s, 3H), 3.19 (s, 3H), 1.45 (s, 9H). The analytical data are consistent with the previously reported characterization. [START_REF] Dai | The Effect of a Trans-Locked Gly-Pro Alkene Isostere on Collagen Triple Helix Stability[END_REF]

Tert-butyl (2-(3,4-dichlorophenyl)-2-oxoethyl)carbamate (3).

According to general procedure TP2, 3,4-dichloro-phenyl magnesium bromide (1.5 eq., 0.5 M, 27.5 mL, 13.7 mmol) was reacted with 2 (1 eq., 2 g, 9.2 mmol) in THF (10 mL). After workup, the crude was purified by silica gel column chromatography (heptane:EtOAc = 5:1) to afford 3 as a white solid (2.4 g, 86%). According to general procedure TP3 (method 1), 3 (1 eq., 1 g, 3.3 mmol) was treated with NaBH4 (4 eq., 0.5 g, 13.2 mmol) in MeOH (30 mL) to afford 4 as a colorless oil (1 g, 99%) without further purification. 1 5). According to general procedure TP4, 4 (1 eq., 500 mg, 1.6 mmol) was reacted with NHPI (1.5 eq., 400 mg, 2.5 mmol), PPh3 (1.8 eq., 771 mg, 2.9 mmol) and DEAD (1.8 eq., 0.46 mL, 2.9 mmol) in THF (15 mL) at rt. The crude was purified by silica gel column chromatography (heptane:EtOAc = 5:1) to afford 5 as a white solid (693 mg, 94%). 1 6). According to general procedure TP5, 5 (1 eq., 400 mg, 0.9 mmol), was reacted with hydrazine hydrate (15 eq., 0.64 mL, 13.3 mmol) in EtOH (15 mL). The crude was purified by silica gel column chromatography (heptane:EtOAc = 2:1) to afford 6 as a white solid (268 mg, 94%). 1 H NMR (400 MHz, CDCl3): δ 7.46 -7.41 (m, 2H), 7.16 (dd, J = 8.2, 1.8 Hz, 1H), 5.44 (s, 2H), 4.86 (s, 1H), 4.60 (dd, J = 7.9, 3.8 Hz, 1H), 3.56 -3.44 (m, 1H), 3.19 (ddd, J = 14.4, 7.9, 4.8 Hz, 1H), 1.43 (s, 9H). [START_REF] Thomas | Toxicity and pharmacokinetics of the nonprotein amino acid l-canavanine in the rat[END_REF] C{ 1 H} NMR (101 MHz, CDCl3): δ 155.9, 139.9, 133.0, 132.2, 130.7, 128.8, 126.2, 84.7, 79.8, 44.9, 28.5.

2-(aminooxy)-2-(3,4-dichlorophenyl)ethanamine dihydrochloride (7).

According to general procedure TP6, 6 (1 eq., 100 mg, 0.31 mmol) was treated with a solution of HCl in dioxane (4 M, 5 mL) to afford 7 as a white solid (88 mg, 96%). 1 H NMR (400 MHz, DMSO-d 6 ): δ 10.48 (s, br, 3H), 8.42 (s, br, 3H), 7.76 -7.72 (m, 2H), 7.46 (dd, J = 8.3, 2.0 Hz, 1H), 5.43 (dd, J = 8.6, 4.0 Hz, 1H), 3.34 -3.17 (m, 2H). [START_REF] Thomas | Toxicity and pharmacokinetics of the nonprotein amino acid l-canavanine in the rat[END_REF] 9). According to general procedure TP7, 7 (1 eq., 50 mg, 0.17 mmol) was dissolved in MeCN (10 mL), and treated successively with TEA (4 eq. 94.6 μL, 0.68 mmol) and a solution of BrCN (1.1 eq., 19.8 mg, 0.19 mmol) in MeCN (0.5 mL) to afford 8 (33.9 mg, 81%) and 9 (7.8 mg, 17%) as white solids after purification by reverse phase C18 column chromatography (MeCN/H2O). Compounds 8 was recrystalized in MeCN at 4 °C. Analytical data for Compound

(S)-tert-butyl (1-oxo-1-phenylpropan-2-yl)carbamate (10).

Step 1: (S)-tert-butyl (1-(methoxy(methyl)amino)-1-oxopropan-2yl)carbamate (36). Following general procedure TP1 and starting from N-Boc-L-alanine (1 eq., 4 g, 21.1 mmol), CDI 1.5 eq., 5.14 g, 31.7 mmol), TEA (1.5 eq, 4.41 mL, 31.7 mmol) and N,Odimethylhydroxylamine hydrochloride (1.5 eq., 3.1 g, 31.7 mmol) in DCM (50 mL), the Weinreb amide of N-Boc-L-alanine 36 was obtained as a white solid (4.67 g, 95%). 1 H NMR (400 MHz, CDCl3): δ 5.25 (d, J = 7.1 Hz, 1H), 4.73 -4.57 (m, 1H), 3.74 (s, 3H), 3.18 (s, 3H), 1.41 (s, 9H), 1.28 (d, J = 6.9 Hz, 3H). The analytical data are consistent with the previously reported characterization. [START_REF] Johnson | Preparation of α-Hydroxy-β-Fmoc Amino Acids from N-Boc Amino Acids[END_REF] Step 2: (S)-tert-butyl (1-oxo-1-phenylpropan-2-yl)carbamate (10). According to general procedure TP2, phenyl magnesium bromide (1.5 eq., 1 M, 12.9 mL, 12.9 mmol) was reacted with the above described Weinreb amide of N-Boc-L-alanine 36 (1 eq., 2 g, 8.6 mmol) in THF (25 mL). After work-up, the crude was purified by silica gel column chromatography (heptane:EtOAc = 10:1) to afford 10 as as a colorless oil (1.93 g, 90%). 1 ). The analytical data are consistent with the previously reported characterization. [START_REF] Lauber | Efficient Aerobic Oxidation of Secondary Alcohols at Ambient Temperature with an ABNO/NOx Catalyst System[END_REF]

(S)-tert-butyl (3-methyl-1-oxo-1-phenylbutan-2-yl)carbamate (11).

Step 1: (S)-tert-butyl (1-(methoxy(methyl)amino)-3-methyl-1oxobutan-2-yl)carbamate (37). Following general procedure TP1 and starting from N-Boc-L-valine (1 eq., 4 g, 18.4 mmol), CDI (1.5 eq., 4.48 g, 27.6 mmol), TEA (1.5 eq, 3.84 mL, 27.6 mmol) and N,O-dimethylhydroxylamine hydrochloride (1.5 eq., 2.69 g, 27.6 mmol) in DCM (50 mL), the Weinreb amide of N-Boc-Lvaline 37 was obtained as a white solid (4.51 g, 94%). 1 H NMR (400 MHz, CDCl3): δ 5.12 (d, J = 9.4 Hz, 1H), 4.55 (t, J = 8.0 Hz, 1H), 3.74 (s, 3H), 3.19 (s, 3H), 1.96 (dq, J = 13.4, 6.7 Hz, 1H), 1.41 (s, 9H), 0.93 (d, J = 6.8 Hz, 3H), 0.88 (d, J = 6.8 Hz, 3H). The analytical data are consistent with the previously reported characterization. [START_REF] King | Defining the Structural Parameters That Confer Anticonvulsant Activity by the Site-by-Site Modification of (R)-N′-Benzyl 2-Amino-3-methylbutanamide[END_REF] Step 2:

(S)-tert-butyl (3-methyl-1-oxo-1-phenylbutan-2yl)carbamate (11). According to general procedure TP2, phenyl magnesium bromide (1.5 eq., 1 M, 11.5 mL, 11.5 mmol) was reacted with the above described Weinreb amide of N-Boc-Lvaline 37 (1 eq., 2 g, 7.7 mmol) in THF (25 mL). After work-up, the crude was purified by silica gel column chromatography (heptane:EtOAc = 10:1) to afford 11 as as a colorless oil (1.88 g, 88 %). 1 H NMR (400 MHz, CDCl3): δ 8.01 -7.87 (m, 2H), 7.63 -7.54 (m, 1H), 7.48 (td, J = 7.5, 1.4 Hz, 2H), 5.43 (d, J = 8.6 Hz, 1H), 5.23 (dd, J = 9.1, 4.1 Hz, 1H), 2.24 -2.03 (m, 1H), 1.45 (s, 9H), 1.03 (d, J = 6.8 Hz, 3H), 0.75 (d, J = 6.8 Hz, 3H). The analytical data are consistent with the previously reported characterization . [START_REF] Garcia | Evaluation of Solution and Solid-Phase Approaches to the Synthesis of Libraries of α,α-Disubstitutedα-acylaminoketones[END_REF]

(S)-tert-butyl 2-benzoylpyrrolidine-1-carboxylate (12).

Step 1: (S)-tert-butyl 2-(methoxy(methyl)carbamoyl)pyrrolidine-1-carboxylate (38). Following general procedure TP1 and starting from N-Boc-L-proline (1 eq., 4 g, 18.6 mmol), CDI (1.5 eq., 4.52 g, 27.9 mmol), TEA (1.5 eq, 3.88 mL, 27.9 mmol) and N,O-dimethylhydroxylamine hydrochloride (1.5 eq., 2.72 g, 27.9 mmol) in DCM (50 mL), the Weinreb amide of N-Boc-L-proline 38 was obtained as a white solid (4.51 g, 94 %). 1 H NMR (400 MHz, CDCl3): δ 4.63 (ddd, J = 41.1, 8.6, 3.4 Hz, 1H), 3.76 (s, 1.5H), 3.69 (s, 1.5H), 3.61 -3.32 (m, 2H), 3.17 (s, 3H), 2.25 -2.08 (m, 1H), 1.99 -1.77 (m, 3H), 1.43 (s, 4.5H), 1.39 (s, 4.5H). The analytical data are consistent with the previously reported characterization. [START_REF] Kong | Rh2(II)-Catalyzed Selective Aminomethylene Migration from Styryl Azides[END_REF] Step 2: (S)-tert-butyl 2-benzoylpyrrolidine-1-carboxylate (12). According to general procedure TP2, phenyl magnesium bromide (1.5 eq., 1 M, 11.6 mL, 11.6 mmol) was reacted with the above described Weinreb amide of N-Boc-L-proline 38 (1 eq., 2 g, 7.7 mmol) in THF (25 mL). After work-up, the crude was purified by silica gel column chromatography (heptane:EtOAc = 10:1) to afford 12 as a colorless oil (1.87 g, 88 %). 1 H NMR (400 MHz, CDCl3): δ 8.04 -7.87 (m, 2H), 7.61 -7.50 (m, 1H), 7.50 -7.40 (m, 2H), 5.39 -5.27 (m, 0.4H), 5.24 -5.14 (m, 0.6H), 3.74 -3.41 (m, 2H), 2.40 -2.18 (m, 1H), 2.01 -1.84 (m, 3H), 1.45 (s, 3.5H), 1.25 (s, 5.5H). The analytical data are consistent with the previously reported characterization. [START_REF] Kong | Rh2(II)-Catalyzed Selective Aminomethylene Migration from Styryl Azides[END_REF] Tert-butyl ((1R,2S)-1-hydroxy-1-phenylpropan-2yl)carbamate (13a). According to general procedure TP3 (method 2), 10 (1 eq., 1 g, 4 mmol) dissolved in THF (10 mL) was treated with DIBAL-H (2 eq., 1 M, 8 mL, 8 mmol) without further purification to afford 13a as a colorless oil (1 g, 99%). 1 H NMR (400 MHz, CDCl3): δ 7.37 -7.26 (m, 5H), 4.85 (d, J = 3.0 Hz, 1H), 4.64 (s, 1H), 4.01 (s, 1H), 3.26 (s, br, 1H), 1.46 (s, 9H), 0.98 (d, J = 7.0 Hz, 3H). The analytical data are consistent with the previously reported characterization. [START_REF] Guo | Synthesis of Chiral Aminophosphines from Chiral Aminoalcohols via Cyclic Sulfamidates[END_REF]

Tert-butyl ((1S,2S)-1-hydroxy-1-phenylpropan-2yl)carbamate (13b).

According to general procedure TP3 (method 3), 10 (1 eq., 1 g, 4 mmol) was dissolved in THF (10 mL) and was treated with borane dimethylsulfide complex (2 eq. , 0.76 mL, 8 mmol) and a solution of (R)-2-Me-CBSoxazaborolidine (0.1 eq. 0.11 g, 4 mmol) in THF (10 mL) to afford 13b as a colorless oil (0.728 g, 71%) after purification by silica gel column chromatography (heptane:EtOAc = 10:1). 1 H NMR (400 MHz, CDCl3): δ 7.35 -7.26 (m, 5H), 4.70 (s, 1H), 4.54 (d, J = 6.0 Hz, 1H), 3.93 -3.79 (m, 1H), 3.27 (s, br, 1H), 1.40 (s, 9H), 1.07 (d, J = 6.8 Hz, 3H). The analytical data are consistent with the previously reported characterization. [START_REF] Lee | Stereoselective Synthesis of Norephedrine and Norpseudoephedrine by Using Asymmetric Transfer Hydrogenation Accompanied by Dynamic Kinetic Resolution[END_REF]

Tert-butyl ((1R,2S)-1-hydroxy-3-methyl-1-phenylbutan-2yl)carbamate (14a).

According to general procedure TP3 (method 2), 11 (1 eq., 1 g, 3.6 mmol) dissolved in THF (10 mL) was treated with DIBAL-H (2 eq., 1 M, 7.2 mL, 7.2 mmol) without further purification to afford 14a as a colorless oil (0.98 g, 97%). ). These data are consistent with the previously reported characterization. [START_REF] Ripa | Discovery of a Novel Oral Glucocorticoid Receptor Modulator (AZD9567) with Improved Side Effect Profile[END_REF]

Tert-butyl ((1S,2S)-1-hydroxy-3-methyl-1-phenylbutan-2yl)carbamate (14b).

According to general procedure TP3 (method 4), 11 (1 eq., 1 g, 3.6 mmol) in THF (10 mL) was added to a suspension of LiAlH4 (2 eq., 1 M, 7.2 mL, 7.2 mmol) to afford 14b as a colorless oil (0.47 g, 47%) after purification by silica gel column chromatography (heptane:EtOAc = 10:1). 1 Hz, 3H). The analytical data are consistent with the previously reported characterization. [START_REF] Schmidt | Stereocontrolled Synthesis of β-Amino Alcohols from Lithiated Aziridines and Boronic Esters[END_REF]

(S)-tert-butyl 2-((R)-hydroxy(phenyl)methyl)pyrrolidine-1carboxylate (15a).

According to general procedure TP3 (method 2), 12 (1 eq., 1 g, 3.6 mmol) in THF (10 mL) was treated with DIBAL-H (2 eq., 1 M, 7.3 mL, 7.3 mmol) without further purification to afford 15a as a colorless oil (1 g, 99%). 1 H NMR (400 MHz, CDCl3): δ 7.37 -7.25 (m, 5H), 5.47 (s, 0.7H), 5.18 (s, 0.3H), 4.89 (s, 0.7H), 4.33 (s, 0.7H), 4.02 (s, 0.3H), 3.58 (s, 0.3H), 3.31 (s, 1H), 2.84 (s, 0.7H), 2.36 -2.14 (m, 0.3H), 1.98 -1.88 (m, 1H), 1.79 (s, 1H), 1.54 (s, 9H), 1.26 -1.16 (m, 1H), 0.95 -0.8 (m, 1H). The analytical data are consistent with the previously reported characterization. [START_REF] Bilke | Catalytic Asymmetric Synthesis of Piperidines from Pyrrolidine: Concise Synthesis of L-733,060[END_REF]

(S)-tert-butyl 2-((S)-hydroxy(phenyl)methyl)pyrrolidine-1carboxylate (15b).

According to general procedure TP3 (Method 1), 12 (1 eq., 1 g, 3.6 mmol) was dissolved in MeOH (30 mL) and treated with NaBH4 (4 eq., 0.55 g, 14.5 mmol) to afford 15b as a colorless oil (0.73 g, 72%). after purification by reverse phase C18 column chromatography (MeOH/H2O). 1 H NMR (400 MHz, CDCl3): δ 7.35 -7.24 (m, 5H), 5.81 (s, 0.7H), 5.41 (s, 0.3H), 5.16 -4.60 (m, 1H), 4.56 -4.21 (m, 1H), 4.12 -4.00 (m, 0.8H), 3.53 -3.16 (m, 2H), 2.79 (s, 0.2H), 1.75 -1.58 (m, 3H), 1.48 (s, 9H). The analytical data are consistent with the previously reported characterization. [START_REF] Bilke | Catalytic Asymmetric Synthesis of Piperidines from Pyrrolidine: Concise Synthesis of L-733,060[END_REF] Tert-butyl ((1S,2S)-1-((1,3-dioxoisoindolin-2-yl)oxy)-1phenylpropan-2-yl)carbamate (16a). According to general procedure TP4, 13a (1 eq., 500 mg, 2 mmol) was reacted with NHPI (1.5 eq., 487 mg, 3 mmol), PPh3 (1.8 eq., 939 mg, 3.6 mmol) and DEAD (1.8 eq., 0.56 mL, 3.6 mmol) in THF (15 mL) at 0°C. The crude was purified by silica gel column chromatography (heptane:EtOAc = 5:1) to afford 16a as a white solid (702 mg, 89%). 1 H NMR (400 MHz, CDCl3): δ 7.76 -7.71 (m, 2H), 7.70 -7.65 (m, 2H), 7.49 -7.43 (m, 2H), 7.37 -7.30 (m, 3H), 5.22 (d, J = 6.5 Hz, 1H), 5.13 (s, 1H), 4.18 (q, J = 7.0 Hz, 1H), 1.42 (s, 9H), 1.20 (d, J = 6.8 Hz, 3H). [START_REF] Thomas | Toxicity and pharmacokinetics of the nonprotein amino acid l-canavanine in the rat[END_REF]

Tert-butyl ((1R,2S)-1-((1,3-dioxoisoindolin-2-yl)oxy)-1phenylpropan-2-yl)carbamate (16b).

According to general procedure TP4, 13b (1 eq., 500 mg, 2 mmol) was reacted with NHPI (2 eq., 649 mg, 4 mmol), PPh3 (2.5 eq., 1.3 g, 5 mmol) and DEAD (2.5 eq., 0.78 mL, 5 mmol) in THF (20 mL) at rt. The crude was purified by silica gel column chromatography (heptane:EtOAc = 5:1) to afford 16b as a colorless oil (757 mg, 96%). 1

Tert-butyl ((1S,2S)-1-((1,3-dioxoisoindolin-2-yl)oxy)-3methyl-1-phenylbutan-2-yl)carbamate (17a).

According to general procedure TP4, 14a (1 eq., 500 mg, 1.8 mmol) was treated with NHPI (1.5 eq., 438 mg, 2.7 mmol), PPh3 (1.8 eq., 845 mg, 3.2 mmol) and DEAD (1.8 eq., 0.51 mL, 3.2 mmol) in THF (15 mL) at 0 °C. The crude was purified by silica gel column chromatography (heptane:EtOAc = 4:1), to afford 17a as a white solid (615 mg, 81%). 1 H NMR (400 MHz, CDCl3): δ 7.72 -7.64 (m, 4H), 7.51 -7.47 (m, 2H), 7.33 -7.27 (m, 3H), 5.52 (d, J = 4.8 Hz, 1H), 5.12 (d, J = 10.2 Hz, 1H), 3.81 (ddd, J = 10.1, 6.8, 4.7 Hz, 1H), 1.98 -1.86 (m, 1H), 1.39 (s, 9H), 1.15 (d, J = 6.7 Hz, 3H), 0.95 (d, J = 6.8 Hz, 3H). [START_REF] Thomas | Toxicity and pharmacokinetics of the nonprotein amino acid l-canavanine in the rat[END_REF]

Tert-butyl ((1S,2S)-1-((1,3-dioxoisoindolin-2-yl)oxy)-3methyl-1-phenylbutan-2-yl)carbamate (17b).

According to general procedure TP4, 14b (1 eq., 400 mg, 1.4 mmol) with was treated NHPI (3 eq., 700 mg, 4.3 mmol), PPh3 (3.5 eq., 1.3 g, 5 mmol) and DEAD (1.8 eq., 0.79 mL, 5 mmol) in THF (15 mL) at 45 °C in an oil bath. The crude was purified by silica gel column chromatography (heptane:EtOAc = 5:1) to afford 17b as a white solid (450 mg, 74%). 1

(S)-tert-butyl 2-((R)-((1,3-dioxoisoindolin-2yl)oxy)(phenyl)methyl)pyrrolidine-1-carboxylate (18a).

According to general procedure TP4, 15a (1 eq., 500 mg, 1.8 mmol) was treated with NHPI (1.5 eq., 441 mg, 2.7 mmol), PPh3 (1.8 eq., 851 mg, 3.2 mmol) and DEAD (1.8 eq., 0.51 mL, 3.2 mmol) in THF (15 mL) at 0 °C. The crude was purified by silica gel column chromatography (heptane:EtOAc = 4:1), to afford 18a as a white solid (716 mg, 94%). 1

(5S,6S)-5-methyl-6-phenyl-5,6-dihydro-4H-1,2,4-oxadiazin-3amine (19a).

Step 1: tert-butyl ((1S,2S)-1-(aminooxy)-1-phenylpropan-2yl)carbamate (39a). According to general procedure TP5, 16a (1 eq., 400 mg, 1 mmol) in EtOH (15 mL) was treated with hydrazine hydrate (15 eq., 0.73 mL, 15.1 mmol). The crude was purified by silica gel column chromatography (heptane:EtOAc = 2:1) to afford tert-butyl ((1S,2S)-1-(aminooxy)-1-phenylpropan-2yl)carbamate 39a as a colorless oil (245 mg, 91%). 1 H NMR (400 MHz, CDCl3): δ 7.40 -7.33 (m, 2H), 7.33 -7.26 (m, 3H), 5.32 (s, 2H), 4.64 -4.53 (m, 1H), 4.49 (d, J = 5.6 Hz, 1H), 3.99 (s, 1H), 1.40 (s, 9H), 1.05 (d, J = 6.8 Hz, 3H). [START_REF] Thomas | Toxicity and pharmacokinetics of the nonprotein amino acid l-canavanine in the rat[END_REF] C{ 1 H} NMR (101 MHz, CDCl3): δ 155.5, 138.6, 128.6, 128.1, 127.5, 89.1, 79.4, 49.9, 28.5, 17.6.

Step 2:

(1S,2S)-1-(aminooxy)-1-phenylpropan-2-amine dihydrochloride (40a). According to general procedure TP6, tertbutyl ((1S,2S)-1-(aminooxy)-1-phenylpropan-2-yl)carbamate 39a (1 eq., 100 mg, 0.38 mmol) was treated with a hydrogen chloride solution in dioxane (4 M, 5 mL) to afford (1S,2S)-1-(aminooxy)-1-phenylpropan-2-amine dihydrochloride 40a as a white solid (89.8 mg, 100%). 1 H NMR (400 MHz, DMSO-d 6 ): δ 10.79 (s, br, 3H), 8.56 (s, br, 3H), 7.57 -7.34 (m, 5H), 5.17 (d, J = 9.1 Hz, 1H), 3.58 (dt, J = 13.6, 6.8 Hz, 1H), 0.93 (d, J = 6.8 Hz, 3H). [START_REF] Thomas | Toxicity and pharmacokinetics of the nonprotein amino acid l-canavanine in the rat[END_REF] (5S,6S)-5-methyl-6-phenyl-5,6-dihydro-4H-1,2,4oxadiazin-3-amine (19a). According to general procedure TP7, the above (1S,2S)-1-(aminooxy)-1-phenylpropan-2-amine dihydrochloride 40a (1 eq., 50 mg, 0.21 mmol) was dissolved in MeCN (10 mL) and treated successively with TEA (4 eq. 116.2 μL, 0.84 mmol) and a solution of BrCN (1.1 eq., 24.4 mg, 0.23 mmol) in MeCN (0.5 mL) to afford the title compound 19a (28.4 mg, 71%) as a white solid after purification by reverse phase C18 column chromatography (MeCN/H2O). Compounds 19a was recrystalized in MeCN at 4 °C. 1 H NMR (400 MHz, DMSOd [START_REF] Eissa | Green Synthesis, Antibacterial, and Antifungal Activities of 1,3,4-Oxadiazines[END_REF] ): δ 7.39 -7.27 (m, 5H), 6.10 (s, 1H), 4.39 (s, 2H), 3.82 (d, J = 8.5 Hz, 1H), 3.40 -3.34 (m, 1H), 0.84 (d, J = 6.3 Hz, 3H). [START_REF] Thomas | Toxicity and pharmacokinetics of the nonprotein amino acid l-canavanine in the rat[END_REF] Step 1: tert-butyl ((1R,2S)-1-(aminooxy)-1-phenylpropan-2yl)carbamate (39b). According to general procedure TP5, 16b (1 eq., 400 mg, 1 mmol) in EtOH (15 mL) was treated with hydrazine hydrate (15 eq., 0.73 mL, 15.1 mmol). The crude was purified by silica gel column chromatography (heptane:EtOAc = 2:1) to afford tert-butyl ((1R,2S)-1-(aminooxy)-1-phenylpropan-2yl)carbamate 39b as a colorless oil (258 mg, 96%). 1 (1R,2S)-1-(aminooxy)-1-phenylpropan-2-amine dihydrochloride (40b). According to general procedure TP6, tertbutyl ((1R,2S)-1-(aminooxy)-1-phenylpropan-2-yl)carbamate 39b (1 eq., 100 mg, 0.38 mmol) was treated with a hydrogen chloride solution in dioxane (4 M, 5 mL) to afford (1R,2S)-1-(aminooxy)-1-phenylpropan-2-amine dihydrochloride 40b as a white solid (88.9 mg, 99%). 1 H NMR (400 MHz, DMSO-d 6 ): δ 10.99 (s, br, 3H), 8.46 (s, br, 3H), 7.51 -7.38 (m, 5H), 5.47 (d, J = 3.8 Hz, 1H), 3.59 (dt, J = 7.0, 3.1 Hz, 1H), 1.08 (d, J = 6.9 Hz, 3H). [START_REF] Thomas | Toxicity and pharmacokinetics of the nonprotein amino acid l-canavanine in the rat[END_REF] tert-butyl ((1S,2S)-1-(aminooxy)-3-methyl-1phenylbutan-2-yl)carbamate (41a). According to general procedure TP5, 17a (1 eq., 400 mg, 0.94 mmol) in EtOH (15 mL) was treated with hydrazine hydrate (15 eq., 0.69 mL, 14.1 mmol). The crude was purified by silica gel column chromatography (heptane:EtOAc = 2:1) to afford tert-butyl ((1S,2S)-1-(aminooxy)-3-methyl-1-phenylbutan-2-yl)carbamate 41a as a white solid (266 mg, 96%). 1 Step 2: (1S,2S)-1-(aminooxy)-3-methyl-1-phenylbutan-2-amine dihydrochloride (42a). According to general procedure TP6, tertbutyl ((1S,2S)-1-(aminooxy)-3-methyl-1-phenylbutan-2yl)carbamate 41a (1 eq., 100 mg, 0.34 mmol) was treated with a hydrogen chloride solution in dioxane (4 M, 5 mL) to afford (1S,2S)-1-(aminooxy)-3-methyl-1-phenylbutan-2-amine dihydrochloride 42a as a white solid (85.3 mg, 94%). 1 H NMR (400 MHz, DMSO-d 6 ): δ 9.02 (s, br, 6H), 7.50 -7.46 (m, 5H), 5.25 (d, J = 9.9 Hz, 1H), 3.44 (dd, J = 9.9, 2.5 Hz, 1H), 1.48 -1.36 (m, 1H), 0.93 (d, J = 7.1 Hz, 3H), 0.81 (d, J = 7.0 Hz, 3H). [START_REF] Thomas | Toxicity and pharmacokinetics of the nonprotein amino acid l-canavanine in the rat[END_REF] (5S,6S)-5-isopropyl-6-phenyl-5,6-dihydro-4H-1,2,4oxadiazin-3-amine (20a). According to general procedure TP7, the above (1S,2S)-1-(aminooxy)-3-methyl-1-phenylbutan-2amine dihydrochloride 42a (1 eq., 50 mg, 0.19 mmol) was dissolved in MeCN (10 mL) and treated successively with TEA (4 eq. 104 μL, 0.75 mmol) and a solution of BrCN (1.1 eq., 21.8 mg, 0.21 mmol) in MeCN (0.5 mL) to afford 20a (30 mg, 73%) as a white solid after purification by reverse phase C18 column chromatography (MeCN/H2O). 1 Step 2: (1R,2S)-1-(aminooxy)-3-methyl-1-phenylbutan-2-amine dihydrochloride (42b). According to general procedure TP6, tertbutyl ((1R,2S)-1-(aminooxy)-3-methyl-1-phenylbutan-2yl)carbamate 41b (1 eq., 100 mg, 0.34 mmol) was treated with a hydrogen chloride solution in dioxane (4 M, 5 mL) to afford (1R,2S)-1-(aminooxy)-3-methyl-1-phenylbutan-2-amine dihydrochloride 42b as a white solid (90.8 mg, 94%). 1 H NMR (400 MHz, DMSO-d 6 ): δ 10.74 (s, br, 3H), 8.15 (s, br, 3H), 7.51 -7.43 (m, 5H), 5.48 (d, J = 5.1 Hz, 1H), 3.51 -3.48 (m, 1H), 1.82 -1.75 (m, 1H), 1.00 (d, J = 6.9 Hz, 3H), 0.97 (d, J = 6.9 Hz, 3H). [START_REF] Thomas | Toxicity and pharmacokinetics of the nonprotein amino acid l-canavanine in the rat[END_REF] Step 3: (5S,6R)-5-isopropyl-6-phenyl-5,6-dihydro-4H-1,2,4oxadiazin-3-amine (20b). According to general procedure TP7, the above (1R,2S)-1-(aminooxy)-3-methyl-1-phenylbutan-2amine dihydrochloride 42b (1 eq., 50 mg, 0.19 mmol) was dissolved in MeCN (10 mL) and treated successively with TEA (4 eq. 104 μL, 0.75 mmol) and a solution of BrCN (1.1 eq., 21.8 mg, 0.21 mmol) in MeCN (0.5 mL) to afford 20b (27.1 mg, 66%) as a white solid after purification by reverse phase C18 column chromatography (MeCN/H2O). 1 (S)-tert-butyl 2-((R)-(aminooxy)(phenyl)methyl)pyrrolidine-1-carboxylate (43). According to general procedure TP5, 18a (1 eq., 400 mg, 0.95 mmol) in EtOH (15 mL) was treated with hydrazine hydrate (15 eq., 0.69 mL, 14.2 mmol). The crude was purified by silica gel column chromatography (heptane:EtOAc = 2:1) to afford (S)-tertbutyl 2-((R)-(aminooxy)(phenyl)methyl)pyrrolidine-1-carboxylate 43 as a colorless oil (269 mg, 97%). 1 was added dropwise and stirred for 5 min. Then a solution of BrCN (1.1 eq., 6.6 mg, 0.062 mmol) in MeCN (0.2 mL) was added dropwise and stirring was maintained for an additional 30 min while the temperature was kept at 0°C. The mixture was concentrated in vacuo and the crude was purified by reverse phase C18 column chromatography (MeOH/H2O) to afford 22 (8.4 mg, 68%) as a white solid. 1 H NMR (400 MHz, DMSO-d 6 ): δ 7.39 -7.30 (m, 5H), 6.05 (s, 2H), 4.54 (d, J = 4.0 Hz, 1H), 3.92 (ddd, J = 7.7, 5.2, 4.1 Hz, 1H), 3.28 -3.12 (m, 2H), 1.81 -1.63 (m, 3H), 1.42 -1.32 (m, 1H). [START_REF] Thomas | Toxicity and pharmacokinetics of the nonprotein amino acid l-canavanine in the rat[END_REF]

Tert-butyl

(3-methyl-2-oxobut-3-en-1-yl)carbamate (26). According to general procedure TP2, 2 (1 eq., 2 g, 9.2 mmol) in THF (10 mL) was treated with isopropenyl magnesium bromide (3.5 eq., 0.5 M, 64.2 mL, 32.1 mmol) to afford 26 as a colorless oil without further purification (1.81 g, 99%). 1 H NMR (400 MHz, CDCl3): δ 5.99 (s, 1H), 5.83 (q, J = 1.4 Hz, 1H), 5.36 (s, 1H), 4.35 (d, J = 4.7 Hz, 2H), 1.90 (t, J = 1.2 Hz, 3H), 1.44 (s, 9H). [START_REF] Thomas | Toxicity and pharmacokinetics of the nonprotein amino acid l-canavanine in the rat[END_REF] C{ 1 H} NMR (101 MHz, CDCl3): δ 196.1, 155.9, 142.5, 125.5, 79.9, 46.7, 28.5, 17.5.

Tert-butyl (2-hydroxy-3-methylbut-3-en-1-yl)carbamate (27).

According to general procedure TP3 (method 5), 26 (1 eq., 1 g, 5 mmol) was dissolved in MeOH (30 mL) and treated with trichlorocerium heptahydrate (1.6 eq., 3 g, 8 mmol) and NaBH4 (1.5 eq., 0.28 g, 7.5 mmol). The crude was purified by silica gel column chromatography (heptane:EtOAc = 2:1) to afford 27 as a colorless oil (0.84 g, 83%). 1 H NMR (400 MHz, CDCl3): δ 5.04 1H), 4.34 (dd, J = 15.9, 7.0 Hz, 1H), 4.22 (dd, J = 15.9, 7.5 Hz, 1H), 3.55 (ddd, J = 12.4, 3.5, 1.2 Hz, 1H), 3.42 (dd, J = 12.4, 10.1 Hz, 1H), 1.81 (t, J = 1.3 Hz, 3H), 1.80 (d, J = 1.3 Hz, 3H), 1.76 (d, J = 1.3 Hz, 3H). HRMS (ESI): m/z calcd. for C11H20N2 15 NO [M + H] + : 211.1577, found: 211.1578. Tert-butyl 3-amino-6-(prop-1-en-2-yl)-5,6-dihydro-2H-1,2,4oxadiazine-2-carboxylate (34). TEA (2 eq., 39.4 μL, 0.283 mmol) was added to a solution of 31 (1 eq., 20 mg, 0.142 mmol) and Boc2O (1.1 eq., 34 mg, 0.156 mmol) in DCM (1 mL). The mixture was stirred at rt for 16 h and concentrated in vacuo. The crude was purified by reverse phase C18 column chromatography (MeOH/H2O) to afford 34 as a white solid (31 mg, 91 %). 1 H NMR (400 MHz, CDCl3): δ 5.32 (s, 2H), 5.03 (q, J = 1.1 Hz, 1H), 5.01 -4.99 (m, 1H), 4.11 (dd, J = 10.3, 3.0 Hz, 1H), 3.81 (dd, J = 12.0, 3.1 Hz, 1H), 3.51 (dd, J = 12.0, 10.3 Hz, 1H), 1.79 (t, J = 1.3 Hz, 3H), 1.50 (s, 9H). [START_REF] Thomas | Toxicity and pharmacokinetics of the nonprotein amino acid l-canavanine in the rat[END_REF] (35). A solution of NaH (4 eq., 4.61 mg, 0.182 mmol) in THF (0.4 mL) was slowly added to a solution of 34 (1 eq., 11 mg, 0.046 mmol) in THF (0.7 mL) under Ar at rt. The mixture was stirred at 60 °C in an oil bath for 1 h and then a solution of prenyl bromide (4 eq., 21.4 μL, 0.182 mmol) in THF (0.1 mL) was added dropwise and stirred at 60 °C in an oil bath for an additional 4 h. The reaction mixture was concentrated in vacuo and purified by reverse phase C18 column chromatography (MeCN/H2O) to afford 35 as a white solid (10.7 mg, 76 %). 1 H NMR (400 MHz, CDCl3): δ 6.65 (s, 1H), 5.34 -5.24 (m, 1H), 5.07 -4.93 (m, 2H), 4.11 (d, J = 10.6 Hz, 1H), 3.81 (dd, J = 11.9, 3.1 Hz, 1H), 3.70 -3.60 (m, 2H), 3.57 -3.48 (m, 1H), 1.81 (s, 3H), 1.72 (s, 3H), 1.66 (s, 3H), 1.51 (s, 9H). [START_REF] Thomas | Toxicity and pharmacokinetics of the nonprotein amino acid l-canavanine in the rat[END_REF] C{ 1 H} NMR (101 MHz, CDCl3): δ 162.3, 150.5, 141.0, 138.9, 120.9, 113.7, 83.7, 76.5, 48.8, 39.9, 29.9, 28.

ASSOCIATED CONTENT

The Supporting Information is available free of charge on the ACS Publications website. NMR studies about BrCN-mediated cyclization, diastereoselective reduction of ketone by DIBAL-H 1 H, [START_REF] Thomas | Toxicity and pharmacokinetics of the nonprotein amino acid l-canavanine in the rat[END_REF] C NMR spectra, Mitsunobu reaction with inversion of configuration, synthetic pathways and NMR characterization of cpd 22, 33, NMR and HRMS comparison between 1 and data from Barrosa et al. [START_REF] Barrosa | Alchornedine, a new anti-trypanosomal guanidine alkaloid from Alchornea glandulosa[END_REF] Experimental procedures for LogD determination and metabolic stabilities. 1 H and [START_REF] Thomas | Toxicity and pharmacokinetics of the nonprotein amino acid l-canavanine in the rat[END_REF] C NMR for all new compounds. FAIR Data is available as Supporting Information for Publication and includes the primary NMR FID files for compounds [1, 8, 19a, 19b, 20a, 20b, 21] Deposition Numbers CCDC 2007599-2007601 contains the supplementary crystallographic data for this paper. These data can be obtained free of charge from The Cambridge Crystallographic Data Centre via www.ccdc.cam.ac.uk/structures

Figure 1 .

 1 Figure 1. Alchornedine, the only natural product reported with a 3-amino-1,2,4-oxadiazine scaffold.

Scheme 1 .

 1 Scheme 1. Synthesis of racemic AOXD 8. X-ray crystallographic structure. Thermal ellipsoids are shown at the 50% probability level.

Figure 2 .

 2 Figure 2. (A-D) Mitsunobu reaction and cyclization to afford AOXDs 19a-b, 20a-b, 21. Yields for 19a, 19b, 20a, 20b, and 21 correspond to the three-step overall yield. (E) X-ray crystallographic structures of 19a and 21. Thermal ellipsoids are shown at the 50% probability level. (F) Ranges of 1 H chemical shift and coupling constant of diastereomeric AOXDs 19-21 in DMSO-d 6 .

Figure 3 .

 3 Figure 3. pKa,log D7.4, and hepatic stability in mouse liver microsomes of AOXDs and its bioisosteres.

Figure 4 .

 4 Figure 4. (A) Analytical comparison between synthesized and isolated alchornedine. (B) Alternative structure for alchornedine. (C) Structures of natural products Alchorneine and Isoalchorneine.

- 1 -

 1 en-2-yl)-5,6-dihydro-4H-1,2,4-oxadiazin-3-amine (1). Tert-butyl 3-((3-methylbut-2-en-1yl)amino)-6-(prop-1-en-2-yl)-5,6-dihydro-2H-1,2,4-oxadiazine-2carboxylate 35 (1 eq., 6.5 mg, 0.021 mmol) was added to a solution of HCl in dioxane (4 M, 1 mL) and stirred at rt for 16 h. The reaction mixture was concentrated in vacuo and basified with a saturated aqueous solution of NaHCO3 until pH reached 8. The crude was concentrated in vacuo and directly purified by reverse phase C18 column chromatography (MeCN/H2O) to afford compound 1 as a white solid (4.2 mg, 96%).1 H NMR (400 MHz, DMSO-d 6 ): δ 5.76 (s, 1H), 5.20 (t, J = 6.9 Hz, 1H), 4.95 (s, 1H), 4.89 (s, 1H), 4.63 -4.57 (m, 1H), 3.72 (dd, J = 9.5, 3.1 Hz, 1H), 3.43 -3.36 (m, 2H), 3.28 -3.24 (m, 1H), 3.01 (t, J = 10.3 Hz, 1H), 1.70 (s, 3H), 1.66 (s, 3H), 1.59 (s, 3H).[START_REF] Thomas | Toxicity and pharmacokinetics of the nonprotein amino acid l-canavanine in the rat[END_REF] C{ 1 H} NMR (101 MHz, DMSO-d 6 ): δ 153.8, 142.5, 132.6, 122.8, 111.9, 74.5, 43.7, 38.8, 25.4, 19.0, 17.7. 1 H NMR (400 MHz, MeOD-d 4 ): δ 5.29 -5.20 (m, 1H), 5.02 (q, J = 1.2 Hz, 1H), 5.01 -4.95 (m, 1H), 3.97 (dd, J = 9.6, 3.3 Hz, 1H), 3.54 (d, J = 6.8 Hz, 2H), 3.40 (dd, J = 11.2, 3.3 Hz, 1H), 3.25 (dd, J = 11.2, 9.6 Hz, 2H), 1.79 (t, J = 1.1 Hz, 3H), 1.73 (d, J = 1.4 Hz, 3H), 1.67 (d, J = 1.3 Hz, 3H). HRMS (ESI): m/z calcd. for C11H20N3O [M + H] + : 210.1606, found: 210.1610.

1 H

 1 NMR (400 MHz, CDCl3): δ 8.04 (d, J = 2.1 Hz, 1H), 7.76 (dd, J = 8.4, 2.1 Hz, 1H), 7.58 (d, J = 8.3 Hz, 1H), 5.45 (s, 1H), 4.60 (d, J = 4.7 Hz, 2H), 1.47 (s, 9H). The analytical data are consistent with the previously reported characterization. 32 Tert-butyl (2-(3,4-dichlorophenyl)-2-hydroxyethyl)carbamate (4).

  H NMR (400 MHz, CDCl3): δ 7.46 (d, J = 2.1 Hz, 1H), 7.40 (d, J = 8.2 Hz, 1H), 7.19 -7.15 (m, 1H), 4.98 (t, J = 6.2 Hz, 1H), 4.79 (dd, J = 7.7, 3.2 Hz, 1H), 3.78 (s, br, 1H), 3.49 -3.38 (m, 1H), 3.26 -3.13 (m, 1H), 1.43 (s, 9H). 13 C{ 1 H} NMR (101 MHz, CDCl3): δ 157.4, 142.3, 132.8, 131.7, 130.5, 128.1, 125.4, 80.5, 73.2, 48.5, 28.5. HRMS (ESI): m/z calcd. for C13H17Cl2NO3 [M + Na] + : 328.0483, found: 328.0492. Tert-butyl (2-(3,4-dichlorophenyl)-2-((1,3-dioxoisoindolin-2yl)oxy)ethyl)carbamate (

  C{ 1 H} NMR (101 MHz, DMSO-d 6 ): δ 136.0, 132.1, 131.4, 131.0, 129.8, 128.1

8 1 H

 1 NMR (400 MHz, DMSO-d 6 ): δ 7.64 -7.60 (m, 2H), 7.35 (dd, J = 8.3, 2.0 Hz, 1H), 6.16 (s, 1H), 4.46 (s, 2H), 4.42 (dd, J = 9.0, 3.2 Hz, 1H), 3.45 (dd, J = 11.4, 3.2 Hz, 1H), 3.15 (dd, J = 11.4, 9.0 Hz, 1H). 13 C{ 1 H} NMR (101 MHz, DMSO-d 6 ): δ 154.1, 140.4, 130.9, 130.4, 130.3, 128.7, 127.0, 72.0, 45.3. HRMS (ESI): m/z calcd. for C9H10Cl2N3O [M + H] + : 246.0201, found: 246.0208. Analytical data for Compound 9 1 H NMR (400 MHz, DMSO-d 6 ): δ 7.72 -7.65 (m, 2H), 7.44 -7.40 (m, 1H), 5.78 (s, 2H), 4.82 (dd, J = 9.5, 3.2 Hz, 1H), 4.10 (dd, J = 11.0, 3.2 Hz, 1H), 3.97 (dd, J = 11.0, 9.5 Hz, 1H).

  H NMR (400 MHz, CDCl3): δ 7.99 -7.94 (m, 2H), 7.61 -7.55 (m, 1H), 7.51 -7.44 (m, 2H), 5.57 (d, J = 7.7 Hz, 1H), 5.35 -5.21 (m, 1H), 1.45 (s, 9H), 1.39 (d, J = 7.1 Hz, 3H

1 H

 1 NMR (400 MHz, CDCl3): δ 7.35 -7.27 (m, 5H), 4.87 (t, J = 4.6 Hz, 1H), 4.35 (d, J = 9.8 Hz, 1H), 3.83 -3.74 (m, 1H), 3.05 (d, J = 4.5 Hz, 1H), 1.78 -1.66 (m, 1H), 1.39 (s, 9H), 1.04 (d, J = 6.7 Hz, 3H), 0.89 (d, J = 6.7 Hz, 3H

  H NMR (400 MHz, CDCl3): δ 7.36 -7.31 (m, 5H), 4.83 -4.76 (m, 2H), 3.52 -3.40 (m, 1H), 2.96 (d, J = 4.4 Hz, 1H), 1.90 -1.79 (m, 1H), 1.36 (s, 9H), 1.01 (d, J = 6.8 Hz, 3H), 0.94 (d, J = 6.8

  H NMR (400 MHz, CDCl3): δ 7.77 -7.72 (m, 2H), 7.71 -7.67 (m, 2H), 7.56 (d, J = 7.5 Hz, 2H), 7.39 -7.27 (m, 3H), 5.43 (s, 1H), 5.40 (s, 1H), 4.06 (s, 1H), 1.43 (s, 9H), 1.24 (d, J = 7.0 Hz, 3H). 13 C{ 1 H} NMR (101 MHz, CDCl3): δ 163.8, 155.4, 136.3, 134.6, 129.0, 128.6, 128.2, 127.6, 123.6, 91.5, 79.5, 51.1, 28.5, 14.2. HRMS (ESI): m/z calcd. for C22H24N2O5 [M + Na] + : 419.1583, found: 419.1590.

  C{ 1 H} NMR (101 MHz, CDCl3): δ 163.7, 156.1, 136.5, 134.5, 129.0, 128.9, 128.3, 128.1, 123.5, 88.2, 79.2, 60.0, 30.6, 28.5, 20.3, 18.5. HRMS (ESI): m/z calcd. for C24H28N2O5 [M + Na] + : 447.1896, found: 447.1902.

  C{ 1 H} NMR (101 MHz, DMSO-d 6 ): δ 153.9, 138.4, 128.2, 128.1, 127.7, 80.9, 50.7, 17.6. HRMS (ESI): m/z calcd. for C10H14N3O [M + H] + : 192.1137, found: 192.1139. (5S,6R)-5-methyl-6-phenyl-5,6-dihydro-4H-1,2,4-oxadiazin-3amine (19b).

  C{1 H} NMR (101 MHz, DMSO-d[START_REF] Eissa | Green Synthesis, Antibacterial, and Antifungal Activities of 1,3,4-Oxadiazines[END_REF] ): δ 134.4, 129.0, 128.7, 126.9, 84.6, 50.0, 12.4.Step 3:(5S,6R)-5-methyl-6-phenyl-5,6-dihydro-4H-1,2,4oxadiazin-3-amine (19b). According to general procedure TP7, the above (1R,2S)-1-(aminooxy)-1-phenylpropan-2-amine dihydrochloride 40b (1 eq., 50 mg, 0.21 mmol) was dissolved in MeCN (10 mL) and treated successively with TEA (4 eq. 116.2 μL, 0.84 mmol) and a solution of BrCN (1.1 eq., 24.4 mg, 0.23 mmol) in MeCN (0.5 mL) to afford 19b (27.6 mg, 69%) as a white solid after purification by reverse phase C18 column chromatography (MeCN/H2O).1 H NMR (400 MHz, DMSO-d 6 ): δ 7.36 -7.22 (m, 5H), 6.25 (s, 1H), 4.55 (d, J = 3.0 Hz, 1H), 4.39 (s, 2H), 3.73 -3.63 (m, 1H), 0.74 (d, J = 6.4 Hz, 3H).[START_REF] Thomas | Toxicity and pharmacokinetics of the nonprotein amino acid l-canavanine in the rat[END_REF] C{ 1 H} NMR (101 MHz, DMSO-d 6 ): δ 154.0, 138.7, 127.9, 126.9, 125.8, 76.3, 48.4, 16.7. HRMS (ESI): m/z calcd. for C10H14N3O [M + H] + : 192.1137, found: 191.1134.

  H NMR (400 MHz, CDCl3): δ 7.35 -7.29 (m, 5H), 5.29 (s, 2H), 4.72 (d, J = 10.4 Hz, 1H), 4.63 (d, J = 4.9 Hz, 1H), 3.63 -3.53 (m, 1H), 1.79 -1.69 (m, 1H), 1.34 (s, 9H), 1.00 (d, J = 6.7 Hz, 3H), 0.90 (d, J = 6.8 Hz, 3H). 13 C{ 1 H} NMR (101 MHz, CDCl3): δ 156.1, 139.7, 128.6, 127.9, 127.0, 86.8, 79.0, 60.0, 30.1, 28.5, 20.3, 18.3.

  H NMR (400 MHz, DMSO-d 6 ): δ 7.37 -7.22 (m, 5H), 6.21 (d, J = 5.0 Hz, 1H), 4.61 (d, J = 3.6 Hz, 1H), 4.37 (s, 2H), 3.42 (dt, J = 5.1, 3.5 Hz, 1H), 1.35 -1.26 (m, 1H), 0.78 (d, J = 6.6 Hz, 3H), 0.70 (d, J = 6.9 Hz, 3H). 13 C{ 1 H} NMR (101 MHz, DMSO-d 6 ): δ 154.3, 138.6, 127.9, 126.9, 125.9, 76.4, 57.3, 28.0, 21.1, 17.1. HRMS (ESI): m/z calcd. for C12H18N3O [M + H] + : 220.1450, found: 220.1453.

  1 H NMR (400 MHz, DMSO-d 6 ): δ 7.41 -7.16 (m, 5H), 4.98 (d, J = 4.3 Hz, 1H), 4.79 (s, 2H), 3.67 (ddd, J = 9.5, 5.5, 4.4 Hz, 1H), 3.32 -3.24 (m, 2H), 1.68 -1.47 (m, 3H), 1.31 -1.08 (m, 1H). 13 C{ 1 H} NMR (101 MHz, DMSO-d 6 ): δ 155.3, 138.7, 127.9, 127.2, 127.1, 75.4, 58.0, 47.4, 27.0, 22.5. HRMS (ESI): m/z calcd. for C12H16N3O [M + H] + : 218.1293, found: 218.1288. (S)-2-((R)-(aminooxy)(phenyl)methyl)pyrrolidine-1carbonitrile (22). To a solution of O-((R)-phenyl((S)-pyrrolidin-2yl)methyl)hydroxylamine dihydrochloride (1 eq., 15 mg, 0.057 mmol) in MeCN (3 mL) at 0 °C, TEA (4 eq., 31.4μL, 0.23 mmol)

  C{ 1 H} NMR (101 MHz, CDCl3): δ 151.8, 150.7, 140.6, 114.2, 84.0, 76.6, 48.4, 28.2, 19.2. HRMS (ESI): m/z calcd. for C11H19N3O3 [M -Boc + H] + : 142.0980, found: 142.0980. Tert-butyl 3-((3-methylbut-2-en-1-yl)amino)-6-(prop-1-en-2yl)-5,6-dihydro-2H-1,2,4-oxadiazine-2-carboxylate

  3, 25.8, 19.2. HRMS (ESI): m/z calcd. for C16H27N3O3 [M -Boc + H] + : 210.1606, found: 210.1615.

Table 1 .

 1 Reduction of ketones 10-12.

  C{ 1 H} NMR (101 MHz, CDCl3): δ 163.7, 155.4, 135.6, 134.5, 129.2, 129.0, 128.4, 128.4, 123.6, 90.9, 79.5, 50.1, 28.5, 17.9. HRMS (ESI): m/z calcd. for C22H24N2O5 [M + Na] + : 419.1590, found: 419.1583.

  H NMR (400 MHz, CDCl3): δ 7.72 -7.67 (m, 2H), 7.67 -7.64 (m, 2H), 7.53 -7.46 (m, 2H), 7.33 -7.26 (m, 3H), 5.36 (d, J = 7.6 Hz, 1H), 4.50 (d, J = 10.7 Hz, 1H), 4.24 -4.15 (m, 1H), 2.32 -2.19 (m, 1H), 1.28 (s, 7H), 1.26 (s, 2H), 1.08 (d, J = 6.8 Hz, 3H), 1.02 (d, J = 6.8 Hz, 3H). 13 C{ 1 H} NMR (101 MHz, CDCl3): δ 163.6, 155.6, 135.8, 134.4, 129.1, 128.9, 128.8, 128.1, 123.5, 89.3, 79.3, 57.7, 28.3, 28.1, 20.6, 16.4. HRMS (ESI): m/z calcd. for C24H28N2O5 [M + Na] + : 447.1896, found: 447.1902.

  C{ 1 H} NMR (101 MHz, DMSO-d 6 ): δ 134.1, 129.7, 128.9, 128.3, 86.4, 49.3, 15.1.

	Step	3:

  H NMR (400 MHz, CDCl3): δ 7.39 -7.32 (m, 2H), 7.31 -7.26 (m, 3H), 5.50 (s, br, 2H), 4.78 -4.65 (m, 1H), 4.61 (d, J = 3.4 Hz, 1H), 4.15 -4.00 (m, 1H), 1.45 (s, 9H), 0.98 (d, J = 7.0 Hz, 3H). 13 C{ 1 H} NMR (101 MHz, CDCl3): δ 155.6, 138.2, 128.5, 127.9, 127.1, 89.1, 79.3, 49.2, 28.6, 15.5.

	Step	2:

  C{ 1 H} NMR (101 MHz, DMSO-d 6 ): δ 129.8, 129.8, 128.9, 128.4, 84.7, 58.1, 27.1, 19.1, 15.1.

	Step	3:

  H NMR (400 MHz, DMSO-d 6 ): δ 7.43 -7.28 (m, 5H), 5.86 (s, 1H), 4.44 (s, 2H), 4.09 (d, J = 8.2 Hz, 1H), 3.34 -3.31 (m, 1H), 1.47 -1.36 (m, 1H), 0.86 (d, J = 7.1 Hz, 3H), 0.79 (d, J = 6.8 Hz, 3H). 13 C{ 1 H} NMR (101 MHz, J = 6.8 Hz, 3H), 0.87 (d, J = 6.8 Hz, 3H). 13 C{ 1 H} NMR (101 MHz, CDCl3): δ 156.1, 138.5, 128.5, 128.0, 127.6, 87.4, 79.1, 57.7, 28.5, 28.4, 20.8, 17.9.

	DMSO-d 6 ): δ 155.0, 138.8, 128.2, 128.0, 127.8, 77.0, 59.6, 28.2,
	19.3, 15.1 HRMS (ESI): m/z calcd. for C12H18N3O [M + H] + :
	220.1450, found: 220.1454.
	(5S,6R)-5-isopropyl-6-phenyl-5,6-dihydro-4H-1,2,4-
	oxadiazin-3-amine (20b).
	Step	1:	tert-butyl	((1R,2S)-1-(aminooxy)-3-methyl-1-
	phenylbutan-2-yl)carbamate (41b). According to general
	procedure TP5, 17b (1 eq., 400 mg, 0.94 mmol) in EtOH (15
	mL) was treated with hydrazine hydrate (15 eq., 0.69 mL, 14.1
	mmol). The crude was purified by silica gel column
	chromatography (heptane:EtOAc = 2:1) to afford tert-butyl
	((1R,2S)-1-(aminooxy)-3-methyl-1-phenylbutan-2-yl)carbamate
	41b as a colorless oil (236 mg, 85%). 1 H NMR (400 MHz,
	CDCl3): δ 7.37 -7.29 (m, 5H), 5.50 (s, 2H), 4.63 (d, J = 5.5 Hz,
	1H), 4.31 (d, J = 10.8 Hz, 1H), 3.96 -3.86 (m, 1H), 1.75 -1.64
	(m, 1H), 1.36 (s, 9H), 1.00 (d,

  C{ 1 H} NMR (101 MHz, DMSO-d 6 ): δ 129.2, 129.2, 128.8, 127.7, 83.5, 58.2, 26.8, 19.7, 17.7.

  H NMR (400 MHz, CDCl3): δ 7.42 -7.27 (m, 5H), 5.43 (s, 2H), 5.01 (d, J = 44.7 Hz, 1H), 4.00 (d, J = 96.9 Hz, 1H), 3.61 -3.26 (m, 2H), 2.13 -1.70 (m, 3H), 1.63 (m, 1H), 1.52 -1.46 (m, 9H). 13 C{ 1 H} NMR (101 MHz, CDCl3): δ 154.6, 140.0, 128.6, 127.6, 126.4, 88.1, 79.7, 62.0, 47.0, 28.7, 25.8, 23.7. hydroxylamine dihydrochloride 44 (1 eq., 50 mg, 0.19 mmol) in MeCN (10 mL) was treated successively with TEA (4 eq. 105 μL, 0.75 mmol) and a solution of BrCN (1.1 eq., 22 mg, 0.21 mmol) in MeCN (0.5 mL) to afford 21 (38.5 mg, 94%) as a white solid after purification by reverse phase C18 column chromatography (MeCN/H2O). Compounds 21 was recrystalized in MeCN at 4 °C.

	Step 2: O-((R)-phenyl((S)-pyrrolidin-2-yl)methyl)hydroxylamine
	dihydrochloride (44). According to general procedure TP6, (S)-
	tert-butyl	2-((R)-(aminooxy)(phenyl)methyl)pyrrolidine-1-
	carboxylate 43 (1 eq., 100 mg, 0.34 mmol) was treated with a
	hydrogen chloride solution in dioxane (4 M, 5 mL) to afford O-
	((R)-phenyl((S)-pyrrolidin-2-yl)methyl)hydroxylamine
	dihydrochloride 44 as a white solid (97.1 mg, 96%). 1 H NMR
	(400 MHz, DMSO-d 6 ): δ 10.53 (s, br, 3H), 9.79 (s, br, 2H), 7.62 -
	7.31 (m, 5H), 5.59 (d, J = 5.0 Hz, 1H), 3.95 -3.76 (m, 1H), 3.24
	-3.06 (m, 2H), 2.04 -1.71 (m, 4H). 13 C{ 1 H} NMR (101 MHz,
	DMSO-d 6 ): δ 135.1, 129.1, 128.8, 127.0, 83.2, 61.7, 45.4, 24.6,
	23.5.
	Step 3: (1R,8aS)-1-phenyl-6,7,8,8a-tetrahydro-1H-pyrrolo[1,2-
	d][1,2,4]oxadiazin-4-amine (21). According to general procedure
	TP7,	the	above	O-((R)-phenyl((S)-pyrrolidin-2-yl)methyl)

  C{ 1 H} NMR (101 MHz, DMSOd 6 ): δ 138.7, 128.1, 127.6, 127.4, 117.0, 86.1, 64.2, 51.2, 26.2, 23.9. HRMS (ESI): m/z calcd. for C12H16N3O [M + H] + : 218.1293, found: 218.1289.

ACKNOWLEDGMENT

We gratefully acknowledge the Ministry of Education of the P.R. China for financial support of this work (PhD grant for S-Q T, China Scholarship Council (201608310124)).

AUTHOR INFORMATION Corresponding Author

Frédéric Bihel -Laboratoire d'Innovation Thérapeutique, UMR7200, CNRS, Université de Strasbourg, Faculty of Pharmacy, 74 Route du Rhin, 67401 Illkirch-Graffenstaden, France ; http://orcid.org/0000-0002-4122-0929; Tel : +33 (0)368854130 ; email: fbihel@unistra.fr

CONFLICTS OF INTEREST

The authors declare no conflict of interest.