Development of Macrocyclic Peptides Containing Epoxyketone with

Article Cite This: J. Med. Chem. 2018, 61, 9177−9204

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Development of Macrocyclic Peptides Containing Epoxyketone with Oral Availability as Proteasome Inhibitors Daqiang Li,† Xiaotuan Zhang,‡,∥ Xiaodong Ma,⊥ Lei Xu,‡,§,∥ Jianjun Yu,† Lixin Gao,‡ Xiaobei Hu,‡ Jiankang Zhang,# Xiaowu Dong,† Jia Li,‡ Tao Liu,*,† Yubo Zhou,*,‡ and Yongzhou Hu*,†

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†

ZJU-ENS Joint Laboratory of Medicinal Chemistry, Zhejiang Province Key Laboratory of Anti-Cancer Drug Research, College of Pharmaceutical Sciences, Zhejiang University, Hangzhou, 310058, People’s Republic of China ‡ National Center for Drug Screening, State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai 201203, China § School of Life Science and Technology, ShanghaiTech University, Shanghai, 201203, China ∥ Graduate School, University of Chinese Academy of Sciences, No. 19A Yuquan Road, 100049 Beijing, China ⊥ Department of Medicinal Chemistry, School of Pharmacy, Anhui University of Chinese Medicine, Hefei 230031, China # Zhejiang University City College, Hangzhou 310015, Zhejiang China S Supporting Information *

ABSTRACT: Macrocyclization has been frequently utilized for optimizing peptide or peptidomimetic-based compounds. In an attempt to obtain potent, metabolically stable, and orally available proteasome inhibitors, 30 oprozomib-derived macrocyclic peptides with structural diversity in their N-terminus and linker were successively designed and synthesized for structure− activity relationship (SAR) studies. As a consequence, the macrocyclic peptides with N-methyl-pyrazole (24p, 24x), imidazole (24t), and pyrazole (24v) as their respective N-termini exhibited favorable in vitro activity and metabolic stability, which translated into their potent in vivo proteasome inhibitory activity after oral administration. In particular, compound 24v, as the most distinguished one among this series, displayed excellent chymotrypsin-like (ChT-L, β5) inhibitory potency (IC50 = 16 nM), low nanomolar antiproliferative activity against all three of the tested cell lines, and superior metabolic stability in mouse liver microsome (MLM), as well as favorable inhibition against ChT-L compared to that of oprozomib in BABL/c mice following po administration at a comparatively low dose, thereby representing a promising candidate for further development.

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INTRODUCTION

comprises three active enzymatic subunits crucial for proteolysis, including caspase-like (C-L, β1), trypsin-like (TL, β2), and chymotrypsin-like (ChT-L, β5) activities.2,10 On the basis of their expression pattern and constitution, CPs in vertebrates can be divided into three types, termed the constitutive proteasome (cCP), the immunoproteasome (iCP), and the thymoproteasome (tCP). Among the proteasomes, cCP, composed of β1c, β2c, and β5c, is ubiquitously expressed. Despite their identical composition (β1i, β2i, and β5t), iCP is predominantly expressed in monocytes and lymphocytes,11 while tCP is exclusively

As the vital component of the ubiquitin-proteasome system (UPS), the proteasome plays an essential role in the degradation of unneeded or damaged proteins1,2 and is responsible for maintaining cellular homeostasis. Additionally, the proteasome mediates a variety of cellular activities, including apoptosis,3 cell cycle regulation,4,5 DNA repair,6 and immune and inflammatory responses.7,8 In contrast, its inhibition leads to the accumulation of unneeded protein, thereby inducing the apoptosis of cells, especially in malignant ones featuring aberrant division and an excess of unwanted intracellular proteins.9 Structurally, the proteasome is a cylindric complex bearing a 20S core particle (20S CP) that contains two outer α rings and two inner β rings. Each β ring © 2018 American Chemical Society

Received: May 23, 2018 Published: September 28, 2018 9177

DOI: 10.1021/acs.jmedchem.8b00819 J. Med. Chem. 2018, 61, 9177−9204

Journal of Medicinal Chemistry

Article

Figure 1. Structures of bortezomib, carfilzomib, ixazomib, and oprozomib.

Figure 2. (a) Crystal structure of human 20S proteasome in complex with oprozomib (green) (PDB 5LEY). (b) Overlay of oprozomib (green) and 24c (cyan) bound to the human 20S proteasome (van der Waals surfaces, PDB 5LEY). Active-site residues participating in polar interactions with the ligand or structural water (red, H2O) are shown as sticks and colored according to atom type (red, oxygen; blue, nitrogen; pink, carbon). Hydrogen bonds are indicated by dashed yellow lines.

distributed in thymic cortical epithelial cells.12 Because of the greater impact on general proteolysis resulting from the inhibition of ChT-L than that of other subunits, massive research endeavors have been made in the exploration of peptidyl proteasome inhibitors (PIs) that interact with ChT-L in light of the binding modes of its endogenous substrates. Upon interaction with the ChT-L site, a majority of these inhibitors adopt an antiparallel β-sheet conformation. At the Cterminus, these peptide inhibitors are commonly equipped with an electrophilic warhead for covalently contacting with Thr1 of the ChT-L.13,14 So far, bortezomib (1), carfilzomib (2), and ixazomib (3), which belong to this class, have been approved for use in humans, which verifies the medicinal potential of the proteasome as an anticancer therapeutic target (Figure 1). Nonetheless, 1, the pioneering PI marketed for treating multiple myeloma (MM) and mantle cell lymphoma (MCL) features a boronate moiety as the warhead, which is capable of interacting with HtrA2/Omi15 and thereby inducing peripheral neuropathy (PN).16,17 Compound 2, introducing an α′,β′epoxyketone as a replacement for the boronate warhead for irreversible interaction with Thr1, has some advantages over 1, including decreased side effects18 and the obviation of bortezomib-mediated drug resistance.19,20 However, both 1 and 2 are routinely intravenously administered due to poor oral bioavailability. To improve the patient compliance, oprozomib (4), an analogue of carfilzomib obtained by peptide

truncation, was discovered and has been advanced into phase I/II clinical trials in patients with hematological malignancy and solid tumors.21,22 In addition, much efforts of medicinal chemistry on 2 delivered PR-89323 and PR-95724 with individual subunit selectivity, respectively. Especially a dual β1i and β5i epoxyketone inhibitor, KZR-616, has successfully completed phase I clinical trials for the treatment of systemic lupus erythematosus with and without nephritis.25 Aside from the boronate and α′β′-epoxyketone, other PIs equipped with aldehydes, vinyl sulfones, vinylamides, α-ketoaldehydes, and βlactones were developed and reviewed in the literature.13 Among them, the most advanced one is marizomib, a derivative of omuralide,26 bearing γ-lactam-β-lactone bicyclic core,27 which is now in Phase III clinical trials for the treatment of newly diagnosed glioblastoma due to the ability of penetrating the blood−brain barrier.28 Another β-lactone derivative, belactosin A and its derivatives,29,30 with generally low cell permeability and limited bioavailability, were further optimized by Shuto and co-workers.31 As a well-established strategy for structural optimization, macrocyclization has been utilized for constraining the conformation of peptide or peptidomimetic-based compounds to reduce the entropy loss correlated with ligand−receptor interaction, enhancing the metabolic stability and improving cellular penetration.32,33 These peptide macrocycles are commonly designed via cyclization between the P1 and P3 or the P2 and P4 sites on the basis of the β-strand geometry of 9178



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Table 1. In Vitro Data of Epoxyketone Macrocycles

a

Mouse, % remaining at 5 min. bMouse, % remaining at 0.5 h. NT = not tested.

proteases.34 In our initial efforts to explore macrocyclic oprozomib derivatives, cyclization between the P1 and P3 was envisioned to be unsuitable because of the potential steric clash with Ala49. Recently, Götz and co-workers reported two macrocyclic peptide PIs bearing aldehyde as the warhead and

an intramolecular linkage between the P2 and P4 side chains.35 Despite the selectivity, the aldehyde functionality was relevant to undesired pharmacokinetics and low oral bioavailability.36 In addition, hydrophobic groups in each amino acid side chain of these were detrimental for gastrointestinal absorption.35 For 9179



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significantly compromised when lowering the dose to 30 mg/kg (Figure 3). To elucidate the underlying factors, a Caco-

the macrocyclic protease inhibitors with intramolecular linkage between N-terminal aromatic rings and the amino acid side chain,34,37,38 synthetic accessibility hindered the introduction of various aryl moieties for systematic structure−activity relationship (SAR) studies and their biological stability, especially oral availability, remain to be investigated. Our group has always been committed to the research of 2 and its derivatives.39,40 Considering the acyclic peptide backbone, 2 and 4, is also prone to hydrolysis by endogenous proteases and peptidases, which compromises its in vivo efficacy.41 Hence, we hypothesized that the strategy of macrocyclization might be beneficial for overcoming this shortcoming and provided a novel series of PIs with favorable potency and oral bioavailability.

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RESULTS AND DISCUSSION To rationally design the macrocyclic peptides appropriate for oral administration, we began by relying on insight from the Xray cocrystal structure of oprozomib bound to the human 20S proteasome42 (Figure 2a, PDB 5LEY). Accordingly, the antiparallel β-sheet backbone of oprozomib was accommodated by the active sites of ChT-L and the β6 subunits with each amide from P1 to P3 engaged in an H-bonding interaction with them, and the α′β′-epoxyketone warhead irreversibly interacted with Thr1, forming a seven-membered 1,4-oxazepane.42 The phenylalanyl moiety at the P1 site was projected into the hydrophobic S1 pocket, while the O-methylseryl moieties at P2 and P3 were located in the S2 and S3 pockets, respectively, without obvious contacts. The thiazole, as the N-terminal moiety, inserted into the lipophilic S4 pocket and conferred hydrophobic interaction with residues Asp125, Pro126, and Ala127 in the β6 subunit. In view of their close spatial proximity, we incorporated a wide variety of chemical linkers between the P2 side chain and the N-terminus for constructing the macrocyclic PIs. To attempt to validate the design rationale, a representative virtual molecule 24c was modeled onto the human 20S proteasome. According to the overlay of 24c and oprozomib in the binding pocket (Figure 2b), 24c maintained the binding mode of oprozomib, and the linker of 24c was involved in additional interaction with Ala22 and Gly23. Additionally, considering the synthetic convenience, phenyl and other six- or five-membered aryl moieties, such as the bioisosteres of thiazole, were introduced to replace this group at the N-terminus. As illustrated in Table 1, compounds 24a−f, bearing 15−20membered phenyl-fused macrocycles, exhibited moderate to potent ChT-L inhibitory activity. Among these, compound 24a−d displayed comparable ChT-L inhibitory activity to that of oprozomib, with IC50 values at the two-digit nanomolar level. Consistent with this result, the four compounds demonstrated attractive cytotoxic activities against both the MM.1S and MV-4-11 cell lines with IC50 values at the twodigit nanomolar level. According to the relatively weakened activity of their 19- and 20-membered macrocyclic counterparts 24e and 24f, the macrocycle of excessive size was detrimental for activity. Compounds 24a−c, with favorable ChT-L inhibitory activity, were therefore selected for evaluating the blood proteasome inhibitory activity in BABL/ c mice following po administration. To our delight, 24c treatment at a dose of 100 mg/kg conferred an equivalent blood proteasome inhibition compared to that of oprozomib treatment at a dose of 30 mg/kg at both 2 and 24 h postadministration. However, the efficacy of 24c was

Figure 3. Mouse oral PD data for oprozomib, 24a−c. CMC = 0.5% CMC-Na; Opr = oprozomib.

2 permeability assay of 24c was performed and it exhibited a moderate membrane permeability ((B−A)/(A−B) ratio (4.4)) (Table 2). Additionally, in the following PK study, 24c Table 2. Caco-2 Permeability of 24ca Caco-2 (×10−6 cm/s) compd

A−B

24c metoprololb atenololb erythromycinb

6.00 ± 0.84 35.19 ± 1.89 0.41 ± 0.07 79.53

26.63 25.94 1.01 15.11

0.47 0.34 0.08 0.58

Average values ± SD of two samples; See Experimental Section for experimental conditions. bLow to high permeable control, respectively. cPapp values were expressed as “95%. HPLC analysis was performed on Agilent 1260 infinity equipped with Cosmosil 5C18-AR-II column (4.6 mm × 250 mm), eluting at 0.8 mL/min using a listed gradient (mobile phase A, H2O; mobile phase B, MeCN) (Table 5).

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CONCLUSION Upon attaching the P2 side chain to the aryl functionality at the N-terminus through a chemical linker, 30 macrocyclic peptides were successively designed and synthesized on the basis of oprozomib for the pursuit of potent, metabolically stable, and orally available proteasome inhibitors. As a result of the SAR investigation focusing on the N-terminus and the chemical linker, four compounds, including 24p, 24x, 24t, and 9185



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which was redissolved in EtOAc and subsequently added cyclohexane or petroleum ether. After removal of the solvent, the corresponding product was afforded. Benzyl (tert-Butoxycarbonyl)-L-serinate (6a). Benzyl bromide (5.6 mL, 46.0 mmol) was slowly added to a mixture of N-Boc-L-serine (5, 8.0 g, 39.0 mmol) and K2CO3 (6.4 g, 46.0 mmol) in DMF (100 mL) at 0 °C. The solution was stirred at room temperature for 20 h and then diluted with EtOAc (200 mL) and H2O (200 mL). the organic layer was separated and washed with saturated NaHCO3 (100 mL × 2) and brine (100 mL × 2), dried over Na2SO4, filtrated, and evaporated under vacuum. The residue was resolved in EtOAc and added PE, filtrated, and washed with additional PE to give 6a (5.2 g, 90%) as a white solid; mp = 68−70 °C. 1H NMR (500 MHz, CDCl3) δ 7.40−7.31 (m, 5H), 5.45 (s, 1H), 5.28−5.16 (m, 2H), 4.48−4.38 (m, 1H), 3.99 (dd, J = 11.0, 3.5 Hz, 1H), 3.92 (dd, J = 11.0, 3.0 Hz, 1H), 1.94 (br s, 1H), 1.45 (s, 9H) ppm. ESI-MS: m/z = 296 [M + H]+. Methyl (tert-Butoxycarbonyl)-L-serinate (6b). To a solution of methyl L-serinate hydrochloride (3.6 g, 30.0 mmol) in MeOH (50 mL), di-tert-butyl dicarbonate (7.9 g, 36 mmol, 1.2 equiv) and TEA (8.3 mL, 60.0 mmol) was added slowly. The mixture was stirred at room temperature for 18 h. The solvent was evaporated under vacuum prior to addition of EtOAc (100 mL) and H2O (100 mL). The organic layer was separated and washed with saturated NH4Cl (80 mL × 2) and brine (80 mL × 2), dried over Na2SO4, filtered, and evaporated under vacuum. The residue was purified by chromatography on silica gel to give 5.9 g of the Boc-L-ser-OMe in 90% yield as a light-yellow oil. 1H NMR (500 MHz, CDCl3) δ 5.47 (s, 1H), 4.43− 4.33 (m, 1H), 3.96 (dd, J = 11.0, 3.5 Hz, 1H), 3.90 (dd, J = 11.0, 3.5 Hz, 1H), 3.78 (s, 3H), 2.27 (br s, 1H), 1.45 (s, 9H) ppm. ESI-MS: m/ z = 220 [M + H]+. Benzyl O-Allyl-N-(tert-butoxycarbonyl)-L-serinate (7a). A three-necked flask was charged with 6a (4.4 g, 15.0 mmol) and Pd(PPh3)4 (0.87 g, 0.75 mmol) under N2. The allylmethyl carbonate (2.3 mL, 21.0 mmol) in anhydrous THF (50 mL) was syringed over 10 min. The reaction mixture was heated to 60 °C for 4 h, upon which TLC (5:1 PE/EtOAc) indicated the loss of starting material. The solvent was removed under reduced pressure, and the residue was diluted with EtOAc (40 mL) and washed with NaHCO3 (40 mL × 2) and brine (40 mL × 2). The organic layer was dried over NaSO4, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography (SiO2; 0−30% EtOAc in PE) to afford 4.1 g (82%) of the product 7a as a clear oil. 1H NMR (500 MHz, CDCl3) δ 7.41−7.30 (m, 5H), 5.86−5.68 (m, 1H), 5.41 (d, J = 8.5 Hz, 1H), 5.32−5.17 (m, 2H), 5.17−5.10 (m, 2H), 4.51−4.42 (m, 1H), 3.99−3.84 (m, 3H), 3.65 (dd, J = 9.5, 3.0 Hz, 1H), 1.45 (s, 9H) ppm. ESI-MS: m/z = 336 [M + H]+. Methyl O-Allyl-N-(tert-butoxycarbonyl)-L-serinate (7b). A three-necked flask was charged with 6b (2.0 g, 9.1 mmol) and Pd(PPh3)4 (0.53 g, 0.46 mmol) under N2. The allylmethyl carbonate (1.5 mL, 12.7 mmol) in anhydrous THF (40 mL) was syringed over 10 min. The reaction mixture was heated to 60 °C for 4 h, upon which TLC indicated the loss of starting material. The solvent was removed under reduced pressure, and the residue was diluted with EtOAc (50 mL) and washed with NaHCO3 (50 mL × 2) and brine (40 mL × 2). The organic layer was dried over NaSO4, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography (SiO2; 0−20% EtOAc in PE) to afford 1.4 g (59%) of the product 7b as a clear oil. 1H NMR (500 MHz, CDCl3) δ 5.87−5.78 (m, 1H), 5.38 (d, J = 8.0 Hz, 1H), 5.27−5.20 (m, 1H), 5.20−5.15 (m, 1H), 4.42 (dt, J = 7.0, 3.0 Hz, 1H), 4.01−3.92 (m, 2H), 3.85 (dd, J = 9.5, 3.0 Hz, 1H), 3.76 (s, 3H), 3.64 (dd, J = 9.5, 3.0 Hz, 1H), 1.45 (s, 9H) ppm. ESI-MS: m/z = 260 [M + H]+. Benzyl N-(tert-Butoxycarbonyl)-O-(hex-5-en-1-yl)-L-serinate (7c). Benzyl bromide (0.79 mL, 6.7 mmol) was slowly added to a mixture of 12 (1.6 g, 5.6 mmol) and K2CO3 (0.92 g, 6.7 mmol) in DMF (15 mL). The solution was stirred at room temperature for 20 h and then diluted with EtOAc (30 mL) and H2O (30 mL). The organic layer was separated and washed with saturated NaHCO3 (20 mL × 2) and brine (20 mL × 2), dried over Na2SO4, filtrated, and

Table 5. Gradient of HPLC t (min)

mobile phase A (%)

mobile phase B (%)

0 5 10 18 19 25 26 28

60 60 40 40 10 10 60 60

40 40 60 60 90 90 40 40

General procedure A for the synthesis of various phenyl, and heteroaryl carboxylates (19a−o, 19t−w and 19y,z) bearing alkene substructure of different length. A mixture of commercial ester (33 mmol, 1.0 equiv), potassium carbonate (66 mmol, 2.0 equiv), and corresponding different length of alkenyl bromide (46 mmol, 1.2 equiv) in 50 mL of DMF was heated to 80 °C for 2 h. TLC analysis indicated that the reaction had ceased. After cooling to room temperature, the solution was diluted with EtOAc (100 mL) and washed with water (100 mL × 2) and brine (50 mL × 2). The organic layer was dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was then purified by flash chromatography on silica gel to give corresponding alkenyl ester. General procedure B for the synthesis of diolefins 21a−q, 21s−z, 21za, and 21zb. The alkenyl ester (6.0 mmol, 1.0 equiv) was dissolved in THF/MeOH (1/1, 8 mL) and aqueous NaOH 2 M (18.0 mmol, 3.0 equiv) was added. The solution was stirred at room temperature for 12 h. The reaction mixture was then acidified to pH 2 with 3 N HCl and extracted with EtOAc (a mixture of EtOAc:acetone = 3:1 for preparing 20t and 20u), washed with brine, dried (Na2SO4), filtered, and concentrated in vacuo to get the crude acid 20a−z and used for further reaction without further purification. The crude acid (1.0 mmol, 1.0 equiv) was dissolved in 5 mL of DCM and cooled to 0 °C. It was sequentially treated with EDCI (1.8 mmol, 1.8 equiv) and HOBt (1.2 mmol, 1.2 equiv). After stirring at room temperature for 30 min, corresponding dipeptide intermediate (1.2 mmol, 1.2 equiv) and DIPEA (3.0 mmol, 3.0 equiv) were added to the reaction mixture and stirred for 30 min, raising the temperature to room temperature, and stirred for 3 h. The solvent was evaporated, and the residue was diluted with 10 mL EtOAc then washed with saturated NH4Cl solution (10 mL × 2), saturated NaHCO3 (10 mL × 2), and brine (10 mL × 2), dried over Na2SO4, filtered, and concentrated in vacuo. Chromatographic purification of the residue provided diolefin derivatives. General procedure C for the synthesis of macrocyclic carboxylic derivatives 22a−x, 22za, and zb. A solution of the diolefin (0.20 mmol, 1.0 equiv) in toluene (0.01 molar) was degassed with dry nitrogen for 15 min. The mixture was stirred for 5 min at 100 °C, after which a degassed solution of Grubbs second-generation catalyst (0.10 equiv) in toluene (0.01 molar) was injected with a syringe for 30 min. The reaction was stirred for half an additional hour. After cooling to room temperature, the solvent was evaporated under reduced pressure and purified by flash chromatography on silica gel. The resulting products (a mixture of geometric isomers) were redissolved in methanol (0.05 molar), stirred overnight with 10% Pd/C (0.10 equiv). The mixture was then filtered over Celite and concentrated to provide the corresponding macrocyclic carboxylic derivatives. General procedure D for the synthesis of target macrocycles 24a−z, 24za, and 24zb. To a solution of macrocyclic carboxylic derivate (0.10 mmol, 1.0 equiv), HOBt (0.12 mmol, 1.2 equiv), and HBTU (0.12 mmol, 1.2 equiv) in 2 mL of THF at 0 °C was added Pheepoxyketone (23, 0.12 mmol, 1.2 equiv) and DIPEA (0.15 mmol, 1.5 equiv). The reaction mixture was stirred at room temperature for 3 h. The solvent was evaporated, and the residue was diluted with 5 mL of EtOAc and then washed with saturated NaHCO3 (5 mL × 2) and brine (5 mL × 2), dried over Na2SO4, filtered, and concentrated in vacuo. Flash chromatography on C18 (50−80% MeOH in H2O) with a flow rate of 25 mL/min yielded the final product as a viscous liquid, 9186



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Hz, 1H), 3.48 (dd, J = 9.0, 7.0 Hz, 1H), 3.39 (s, 3H), 1.45 (s, 9H) ppm. 13C NMR (100 MHz, CDCl3) δ 170.54, 169.14, 155.60, 134.10, 117.35, 80.07, 72.37, 72.29, 69.56, 68.58, 59.28, 59.09, 53.12, 28.35. ESI-MS: m/z = 361 [M + H]+. Benzyl N-(N-(tert-Butoxycarbonyl)-O-methyl-L -seryl)-O(hex-5-en-1-yl)-L-serinate (9c). To a solution of 7c (0.91 g, 2.4 mmol) in 4 N HCl−dioxane (8 mL) was stirred at room temperature for 2 h. The volatiles were removed in vacuo. The residual HCl was further removed by adding Et2O (5 mL × 2) to the hydrochloride salt followed by its removal in vacuo. This residue 8c and Boc-O-methylL-serine (0.44 g, 2.0 mmol) were dissolved in THF (6 mL). The mixture was then treated sequentially at 0 °C with DIPEA (1.0 mL, 6.0 mmol), HOBt (0.32 g, 2.4 mmol), and EDCI (0.69 g, 3.6 mmol). The mixture was stirred for 5 h at room temperature. The solvent was evaporated in vacuum, and the residue was diluted by EtOAc (10 mL) and H2O (10 mL). The organic layer was separated and washed with saturated NaHCO3 (8 mL × 2) and brine (8 mL × 1), dried over Na2SO4, filtered, and concentrated in vacuo. Chromatography provided 9c (0.56 g, 59%) as a viscous liquid. 1H NMR (500 MHz, CDCl3) δ 7.41−7.29 (m, 6H), 5.83−5.72 (m, 1H), 5.42 (s, 1H), 5.26−5.11 (m, 2H), 5.03−4.92 (m, 2H), 4.73 (dt, J = 8.0, 3.0 Hz, 1H), 4.28 (s, 1H), 3.88 (dd, J = 9.5, 3.0 Hz, 1H), 3.77 (dd, J = 9.0, 4.0 Hz, 1H), 3.64 (dd, J = 9.5, 3.0 Hz, 1H), 3.46−3.32 (m, 6H), 2.07− 1.99 (m, 2H), 1.56−1.48 (m, 2H), 1.45 (s, 9H), 1.41−1.33 (m, 2H) ppm. ESI-MS: m/z = 479 [M + H]+. Benzyl (S)-2-((S)-2-((tert-Butoxycarbonyl)amino)-3methoxypropanamido)hex-5-enoate (9d). To a solution of 7d (0.77 g, 2.4 mmol) in 4 N HCl−dioxane (5 mL) was stirred at room temperature for 2 h. The volatiles were removed in vacuo. The residual HCl was further removed by adding Et2O (3 mL × 2) to the hydrochloride salt followed by its removal in vacuo. The crude product 8d was used in the next step without further purification. Boc-O-methyl-serine (0.44 g, 2.0 mmol) was dissolved in 6 mL of DCM and cooled to 0 °C. It was sequentially treated with EDCI (0.69 g, 3.6 mmol) and HOBt (0.32 g, 2.4 mmol). After stirring at room temperature for 30 min, amine hydrochloride 8d and DIPEA (1.0 mL, 6.0 mmol) were added to the reaction mixture and stirred for 30 min, raising the temperature to room temperature, and stirred for 6 h. The solvent was evaporated. The residue was diluted with 20 mL of EtOAc and then washed with saturated NH4Cl solution (15 mL × 2), saturated NaHCO3 (15 mL × 2) and brine (15 mL × 2), dried over Na2SO4, filtered, and concentrated in vacuo. Chromatographic purification of the residue provided compounds 9d (0.48g, 57%) as a viscous liquid. 1H NMR (500 MHz, CDCl3) δ 7.40−7.31 (m, 5H), 7.11 (d, J = 5.5 Hz, 1H), 5.79−5.68 (m, 1H), 5.41 (d, J = 3.5 Hz, 1H), 5.17 (q, J = 12.0 Hz, 2H), 5.01−4.94 (m, 2H), 4.67 (td, J = 7.5, 4.5 Hz, 1H), 4.25 (s, 1H), 3.79 (dd, J = 9.0, 4.0 Hz, 1H), 3.46−3.40 (m, 1H), 3.35 (s, 3H), 2.11−1.92 (m, 3H), 1.84−1.74 (m, 1H), 1.45 (s, 9H) ppm. 13C NMR (100 MHz, CDCl3) δ 171.83, 170.29, 155.56, 136.97, 135.37, 128.69, 128.55, 128.37, 115.80, 80.33, 72.09, 67.21, 59.09, 53.75, 52.03, 31.66, 29.34, 28.36. ESI-MS: m/z = 421 [M + H]+. Benzyl O-Allyl-N-(O-methyl-L-seryl)-L-serinate Hydrochloride (10a). To a solution of 9a (2.2 g, 5.0 mmol) in 4 N HCl−dioxane (10 mL) was stirred at room temperature for 2 h. The volatiles were removed in vacuo. The residual HCl was further removed by adding Et2O (5 mL × 2) to the hydrochloride salt followed by its removal in vacuo. The crude product was used in the next step without further purification. 13C NMR (100 MHz, DMSO) δ 169.31, 166.74, 135.71, 134.56, 128.38, 128.07, 127.77, 117.07, 71.37, 70.42, 68.86, 66.29, 58.48, 52.72, 51.88. ESI-MS: m/z = 337 [M + H]+. Methyl O-Allyl-N-(O-methyl-L-seryl)-L-serinate Hydrochloride (10b). To a solution of 9b (1.8 g, 5.0 mmol) in 4 N HCl− dioxane (10 mL) was stirred at room temperature for 2 h. The volatiles were removed in vacuo. The residual HCl was further removed by adding Et2O (5 mL × 2) to the hydrochloride salt followed by its removal in vacuo. The crude product 10b was used in the next step without further purification. 13C NMR (100 MHz, DMSO) δ 170.78, 166.57, 134.73, 116.99, 71.37, 70.57, 69.15, 58.51, 52.77, 52.72, 52.24, 52.09. ESI-MS: m/z = 261 [M + H]+.

evaporated under vacuum to give 7c (1.1 g, 52%) as a viscous liquid. 1 H NMR (500 MHz, CDCl3) δ 7.41−7.29 (m, 5H), 5.85−5.71 (m, 1H), 5.38 (d, J = 8.5 Hz, 1H), 5.28−5.09 (m, 2H), 5.05−4.89 (m, 2H), 4.52−4.39 (m, 1H), 3.84 (dd, J = 9.5, 3.0 Hz, 1H), 3.63 (dd, J = 9.5, 3.0 Hz, 1H), 3.45−3.27 (m, 2H), 2.07−1.99 (m, 2H), 1.55−1.33 (m, 13H) ppm. ESI-MS: m/z = 378 [M + H]+. Benzyl (S)-2-((tert-Butoxycarbonyl)amino)hex-5-enoate (7d). To a flask containing activated zinc (0.33 g, 4.9 mmol) at 0 °C under N2 was added dropwise a solution of 13 (0.50 g, 1.2 mmol) in DMF. The reaction mixture was removed from ice bath and allowed to stir at room temperature for 3 h, upon which TLC (1:4 EtOAc in PE) indicated the loss of starting material and formation of a lower Rf spot. At this point, the reaction mixture was stopped to let the solid settle to the bottom. The supernatant was then carefully transferred by syringe to a suspension of copper(I) bromide (36 mg, 0.25 mmol) in DMF (2 mL) at −15 °C that also contained allyl chloride (0.22 g, 1.9 mmol). After complete addition, the cooling bath was removed and stirring was continued overnight. At this point, EtOAc (10 mL) was added to the reaction mixture and stirred for 15 min washed with H2O (10 mL × 2), 1N Na2S2O3 (20 mL × 2), and brine (20 mL × 1), dried over Na2SO4, filtrated, and concentrated in vacuo. The crude residue was purified by flash chromatography (SiO2, 1:8 EtOAc in PE) to yield 0.23 g (60%) of 7d as a colorless oil. 1H NMR (500 MHz, CDCl3) δ 7.41−7.31 (m, 5H), 5.81−5.70 (m, 1H), 5.24−5.10 (m, 2H), 5.10−4.93 (m, 3H), 4.42−4.32 (m, 1H), 2.15− 2.02 (m, 2H), 1.97−1.82 (m, 1H), 1.78−1.67 (m, 1H), 1.44 (s, 9H) ppm. ESI-MS: m/z = 320 [M + H]+. Benzyl O-Allyl-N-(N-(tert-butoxycarbonyl)-O-methyl- L seryl)-L-serinate (9a). To a solution of 7a (4.1 g, 12.0 mmol) in 4 N HCl−dioxane (24 mL) was stirred at room temperature for 2 h. The volatiles were removed in vacuo. The residual HCl was further removed by adding Et2O (12 mL × 2) to the hydrochloride salt followed by its removal in vacuo. This residue 8a and Boc-O-methylL-serine (2.2 g, 10.0 mmol) were dissolved in THF (30 mL). The mixture was then treated sequentially at 0 °C with DIPEA (2.5 mL, 15.0 mmol), HOBt (1.6 g, 12.0 mmol), and EDCI (3.5 g, 18.0 mmol). The mixture was stirred for 5 h at room temperature. The solvent was evaporated in vacuum, and the residue was diluted by EtOAc (50 mL) and H2O (50 mL). The organic layer was separated and washed with saturated NaHCO3 (50 mL × 2) and brine (30 mL × 1), dried over Na2SO4, filtered, and concentrated in vacuo. Chromatography (10% EtOAc−hexanes) provided 9a (2.7 g, 61%) as a viscous liquid. 1H NMR (500 MHz, CDCl3) δ 7.39 (d, J = 6.5 Hz, 1H), 7.36−7.29 (m, 5H), 5.80−5.70 (m, 1H), 5.43 (d, J = 5.0 Hz, 1H), 5.26−5.21 (m, 1H), 5.21−5.16 (m, 1H), 5.16−5.11 (m, 2H), 4.74 (dt, J = 8.0, 3.0 Hz, 1H), 4.31−4.24 (m, 1H), 3.94−3.87 (m, 3H), 3.77 (dd, J = 9.0, 4.0 Hz, 1H), 3.65 (dd, J = 9.5, 3.0 Hz, 1H), 3.43 (dd, J = 9.0, 7.0 Hz, 1H), 3.33 (s, 3H), 1.44 (s, 9H) ppm. 13C NMR (100 MHz, CDCl3) δ 170.49, 169.79, 155.48, 135.44, 134.03, 128.58, 128.38, 128.16, 117.30, 80.17, 72.24, 72.15, 69.60, 67.24, 59.01, 53.67, 53.07, 28.32. ESI-MS: m/z = 437 [M + H]+. Methyl O-Allyl-N-(N-(tert-butoxycarbonyl)-O-methyl- L seryl)-L-serinate (9b). To a solution of 7b (3.1 g, 12.0 mmol) in 4 N HCl−dioxane (24 mL) was stirred at room temperature for 2 h. The volatiles were removed in vacuo. The residual HCl was further removed by adding Et2O (12 mL × 2) to the hydrochloride salt followed by its removal in vacuo. This residue 8b and Boc-O-methylL-serine (2.2 g, 10.0 mmol) were dissolved in THF (30 mL). The mixture was then treated sequentially at 0 °C with DIPEA (2.5 mL, 15.0 mmol), HOBt (1.6 g, 12.0 mmol), and EDCI (3.5 g, 18.0 mmol). The mixture was stirred for 5 h at room temperature. The solvent was evaporated in vacuum, and the residue was diluted by EtOAc (50 mL) and H2O (50 mL). The organic layer was separated and washed with saturated NaHCO3 (50 mL × 2) and brine (30 mL × 1), dried over Na2SO4, filtered, and concentrated in vacuo. Chromatography (10% EtOAc−PE) provided 9b (2.6 g, 72%) as a viscous liquid. 1H NMR (500 MHz, CDCl3) δ 7.34 (d, J = 3.5 Hz, 1H), 5.87−5.76 (m, 1H), 5.42 (s, 1H), 5.26−5.15 (m, 2H), 4.70 (dt, J = 8.0, 3.5 Hz, 1H), 4.31−4.24 (m, 1H), 3.99−3.95 (m, 2H), 3.88 (dd, J = 9.5, 3.0 Hz, 1H), 3.81 (dd, J = 9.0, 3.5 Hz, 1H), 3.76 (s, 3H), 3.65 (dd, J = 9.5, 3.5 9187



Article

Benzyl O-(Hex-5-en-1-yl)-N-(O-methyl-L-seryl)-L-serinate Hydrochloride (10c). To a solution of 9c (0.48 g, 1.0 mmol) in 4 N HCl−dioxane (2 mL) was stirred at room temperature for 2 h. The volatiles were removed in vacuo. The residual HCl was further removed by adding Et2O (2 mL × 2) to the hydrochloride salt followed by its removal in vacuo. The crude product 10c was used in the next step without further purification. 13C NMR (100 MHz, DMSO) δ 169.40, 166.75, 138.62, 135.76, 128.43, 128.04, 127.85, 114.80, 70.62, 70.53, 66.38, 62.97, 58.49, 52.85, 52.11, 32.94, 28.37, 24.74. ESI-MS: m/z = 379 [M + H]+. Benzyl (S)-2-((S)-2-Amino-3-methoxypropanamido)hex-5enoate Hydrochloride (10d). To a solution of 9d (0.42 g, 1.0 mmol) in 4 N HCl−dioxane (2 mL) was stirred at room temperature for 2 h. The volatiles were removed in vacuo. The residual HCl was further removed by adding Et2O (2 mL × 2) to the hydrochloride salt followed by its removal in vacuo. The crude product 10d was used in the next step without further purification. ESI-MS: m/z = 321 [M + H]+. N-(tert-Butoxycarbonyl)-O-(hex-5-en-1-yl)- L-serine (12). (tert-Butoxycarbonyl)-L-serine (2.1 g, 10.0 mmol) was dissolved in DMF (25 mL). The solution was cooled to 0 °C (ice bath) before sodium hydride (60% dispersion in mineral oil, 0.60 g, 25.0 mmol) was added slowly under stirring. After gas evolution had ceased, 6bromohex-1-ene (1.3 mL, 10.0 mmol) was added dropwise and the reaction mixture stirred for 18 h at room temperature. The reaction mixture was cooled to 0 °C and diluted with EtOAc (50 mL) and H2O (50 mL), and the organic layer was separated and washed with H2O (50 mL × 2), and brine (50 mL × 2), dried over Na2SO4, filtrated, and evaporated under reduced pressure to yield 12 (1.6 g, 56%) as a viscous liquid. 1H NMR (500 MHz, CDCl3) δ 5.84−5.72 (m, 1H), 5.42 (d, J = 8.0 Hz, 1H), 5.06−4.88 (m, 2H), 4.47−4.37 (m, 1H), 3.87 (dd, J = 11.0, 3.0 Hz, 2H), 3.65 (dd, J = 9.5, 3.5 Hz, 1H), 3.45 (t, J = 6.5 Hz, 2H), 2.08−2.02 (m, 2H), 1.60−1.52 (m, 2H), 1.49−1.38 (m, 11H) ppm. 13C NMR (100 MHz, CDCl3) δ 174.39, 156.00, 138.72, 114.72, 80.39, 71.64, 70.38, 54.00, 33.51, 28.84, 28.42, 25.32. ESI-MS: m/z = 288 [M + H]+. Benzyl (R)-2-((tert-Butoxycarbonyl)amino)-3-iodopropanoate (13). A flask was charged with benzyl (tert-butoxycarbonyl)L-serinate (6a, 3.0 g, 10.0 mmol) and PPh3 (3.9 g, 15.0 mmol) under N2. To this was added anhydrous THF (30 mL), and the solution was cooled to 0 °C by immersion in an ice bath. Pyridine (1.7 mL, 20.0 mmol) was added followed by solid iodine (3.8 g, 15.0 mmol) in three portions at 0 °C. the reaction mixture was warmed to room temperature and stirred for 5 h, diluted with Et2O (40 mL) and washed with 1 N HCl (20 mL × 2), Na2S2O3 (20 mL × 2), and brine (20 mL × 2), and dried over Na2SO4, filtrated, and concentrated in vacuo. Purified with chromatography on silica gel (1:60 to 1:30 EtOAc in PE) to yield the white solid (3.6 g, 89%); mp = 79−80 °C. 1 H NMR (500 MHz, CDCl3) δ 7.41−7.33 (m, 5H), 5.36 (d, J = 7.0 Hz, 1H), 5.21 (q, J = 12.0 Hz, 2H), 4.59−4.51 (m, 1H), 3.61 (dd, J = 10.0, 4.0 Hz, 1H), 3.56 (dd, J = 10.0, 4.0 Hz, 1H), 1.45 (s, 9H) ppm. ESI-MS: m/z = 406 [M + H]+. General Procedure for Compounds 14a, 14b Synthesis. (tert-Butoxycarbonyl)-L-serine (5, 2.1 g, 10.0 mmol) was dissolved in DMF (30 mL). The solution was cooled to 0 °C (ice bath) before sodium hydride (60% dispersion in mineral oil, 0.60 g, 25.0 mmol) was added slowly under stirring. After gas evolution had ceased, 2-(2(benzyloxy)ethoxy)ethyl 4-methylbenzenesulfonate or 2-(2-(2(benzyloxy)ethoxy) ethoxy)ethyl 4-methylbenzenesulfonate (10.0 mmol, 1.0 equiv) was added dropwise and the reaction mixture stirred for 18 h at room temperature. The reaction mixture was cooled to 0 °C and diluted with EtOAc (50 mL) and H2O (50 mL), and the organic layer was separated and washed with H2O (50 mL × 2) and brine (50 mL × 2), dried over Na2SO4, filtrated, and evaporated under reduced pressure to yield acid. O-(2-(2-(Benzyloxy)ethoxy)ethyl)-N-(tert-butoxycarbonyl)-L-serine (14a). Viscous liquid (2.0 g, 52%). 1H NMR (500 MHz, CDCl3) δ 7.36−7.31 (m, 4H), 7.31−7.27 (m, 1H), 5.60 (d, J = 7.0 Hz, 1H), 4.62−4.54 (m, 2H), 4.46−4.38 (m, 1H), 3.96 (dd, J = 9.5, 3.0 Hz, 1H), 3.77−3.56 (m, 9H), 1.44 (s, 9H) ppm. 13C NMR (100 MHz,

CDCl3) δ 173.53, 155.96, 137.92, 128.45, 127.92, 127.78, 80.22, 73.25, 71.05, 70.85, 70.49, 70.28, 69.34, 53.94, 28.36. ESI-MS: m/z = 384 [M + H]+. (S)-13-((tert-Butoxycarbonyl)amino)-1-phenyl-2,5,8,11-tetraoxatetradecan-14-oic Acid (14b). Viscous liquid (3.0 g, 71%). 1H NMR (500 MHz, CDCl3) δ 7.38−7.27 (m, 5H), 5.56 (d, J = 8.0 Hz, 1H), 4.66−4.56 (m, 2H), 4.45−4.40 (m, 1H), 3.94 (dd, J = 9.5, 3.0 Hz, 1H), 3.76−3.55 (m, 13H), 1.45 (s, 9H) ppm. 13C NMR (100 MHz, CDCl3) δ 173.06, 155.82, 137.87, 128.38, 127.88, 127.71, 79.95, 73.17, 71.08, 70.76, 70.47, 70.45, 70.38, 70.11, 69.26, 53.97, 28.31. ESI-MS: m/z = 428 [M + H]+. General Procedure for Compounds 16a, 16b Synthesis. To a solution of ether acid (5.2 mmol) in MeOH (25 mL) at 0 °C was added thionyl chloride (0.83 mL, 11.0 mmol, 2.1 equiv) dropwise over a 5 min period. The mixture was stirred at 60 °C for 2 h and then concentrated under high vacuum to give the crude product for next step without further purification. The Boc-O-methyl-serine (1.1 g, 5.0 mmol) was dissolved in 15 mL of DCM and cooled to 0 °C. It was sequentially treated with EDCI (1.7 g, 9.0 mmol) and HOBt (0.68 g, 5.0 mmol). After stirring at room temperature for 30 min, amine hydrochloride described above (6.0 mmol) and DIPEA (2.5 mL, 15.0 mmol) were added to the reaction mixture, raising the temperature to room temperature, and stirred for 6 h. The solvent was evaporated. The residue was diluted with 20 mL of EtOAc and then washed with saturated NH4Cl solution (15 mL × 2), saturated NaHCO3 (15 mL × 2), and brine (15 mL × 2), dried over Na2SO4, filtered, and concentrated in vacuo. Chromatographic purification of the residue provided the corresponding compound. Methyl O-(2-(2-(Benzyloxy)ethoxy)ethyl)-N-(N-(tert-butoxycarbonyl)-O-methyl-L-seryl)-L-serinate (16a). Viscous liquid (two steps, 1.3 g, 51%). 1H NMR (500 MHz, CDCl3) δ 7.39−7.27 (m, 6H), 5.50 (d, J = 4.5 Hz, 1H), 4.74−4.66 (m, 1H), 4.57 (s, 2H), 4.34−4.25 (m, 1H), 3.96 (dd, J = 10.0, 3.5 Hz, 1H), 3.79 (dd, J = 9.0, 4.0 Hz, 1H), 3.75−3.70 (m, 4H), 3.67−3.59 (m, 8H), 3.48 (dd, J = 9.0, 6.5 Hz, 1H), 3.37 (s, 3H), 1.46 (s, 9H) ppm. ESI-MS: m/z = 499 [M + H]+. Methyl (S)-13-((S)-2-((tert-Butoxycarbonyl)amino)-3-methoxypropanamido)-1-phenyl-2,5,8,11-tetraoxatetradecan-14-oate (16b). Viscous liquid (two steps, 1.2 g, 43%). 1H NMR (500 MHz, CDCl3) δ 7.40−7.31 (m, 5H), 7.31−7.27 (m, 1H), 5.48 (d, J = 4.0 Hz, 1H), 4.70 (dt, J = 7.0, 3.0 Hz, 1H), 4.57 (s, 2H), 4.33−4.25 (m, 1H), 3.94 (dd, J = 10.0, 3.5 Hz, 1H), 3.80 (dd, J = 9.5, 3.5 Hz, 1H), 3.74 (s, 3H), 3.71 (dd, J = 9.5, 3.0 Hz, 1H), 3.69−3.58 (m, 12H), 3.49 (dd, J = 9.2, 6.6 Hz, 1H), 3.38 (s, 3H), 1.46 (s, 9H) ppm. 13C NMR (100 MHz, CDCl3) δ 170.70, 170.53, 155.60, 138.34, 128.46, 127.84, 127.71, 80.16, 73.33, 72.67, 72.37, 72.25, 71.07, 70.86, 70.71, 70.48, 69.53, 59.14, 53.05, 52.61, 28.41. ESI-MS: m/z = 543 [M + H]+. General Procedure for Hydroxyl Compounds 17a, 17b Synthesis. A solution of ester (16a or 16b, 1.4 mmol, 1.0 equiv) in MeOH (17.1 mL) was treated with 10% Pd/C (120 mg) and hydrogenated under balloon pressure at room temperature. After stirring for 3 h at room temperature, the insoluble materials were separated by filtration through Celite and eluting with MeOH. The filtrate was then concentrated in vacuo. Concentration of the filtrate yielded hydroxyl compound. Methyl N-(N-(tert-Butoxycarbonyl)-O-methyl-L-seryl)-O-(2-(2hydroxyethoxy)ethyl)-L-serinate (17a). Viscous liquid (0.59 g, 100%). 1H NMR (500 MHz, CDCl3) δ 7.43 (d, J = 8.0 Hz, 1H), 5.64 (d, J = 4.0 Hz, 1H), 4.73 (dt, J = 8.0, 3.0 Hz, 1H), 4.35−4.28 (m, 1H), 3.98 (dd, J = 10.0, 3.0 Hz, 1H), 3.81 (dd, J = 9.0, 4.0 Hz, 1H), 3.78−3.70 (m, 6H), 3.66−3.57 (m, 6H), 3.50 (dd, J = 9.5, 6.0 Hz, 1H), 3.38 (s, 3H), 1.45 (s, 9H) ppm. 13C NMR (100 MHz, CDCl3) δ 170.75, 170.53, 155.67, 80.34, 72.72, 72.49, 71.19, 70.90, 70.29, 61.80, 59.24, 53.15, 52.75, 28.44. ESI-MS: m/z = 409 [M + H]+. Methyl N-(N-(tert-Butoxycarbonyl)-O-methyl-L-seryl)-O-(2-(2-(2hydroxyethoxy) ethoxy)ethyl)-L-serinate (17b). Viscous liquid (0.60 g, 95%). 1H NMR (500 MHz, CDCl3) δ 7.83 (d, J = 7.0 Hz, 1H), 5.68 (d, J = 6.0 Hz, 1H), 4.79−4.70 (m, 1H), 4.42−4.32 (m, 1H), 9188



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3.96 (dd, J = 9.5, 3.0 Hz, 1H), 3.80−3.54 (m, 18H), 3.38 (s, 3H), 2.45 (br s, 1H), 1.45 (s, 9H) ppm. ESI-MS: m/z = 453 [M + H]+. Methyl 2-(Allyloxy)benzoate (19a). Obtained following the general procedure A from commercial methyl 2-hydroxybenzoate 18a. Colorless oil (5.9 g, 93%). 1H NMR (500 MHz, CDCl3) δ 7.80 (dd, J = 8.0, 2.0 Hz, 1H), 7.44 (ddd, J = 8.0, 7.5, 2.0 Hz, 1H), 7.01−6.94 (m, 2H), 6.11−6.02 (m, 1H), 5.54−5.48 (m, 1H), 5.32−5.27 (m, 1H), 4.63 (dt, J = 5.0, 1.5 Hz, 2H), 3.90 (s, 3H) ppm. ESI-MS: m/z = 193 [M + H]+. Methyl 2-(But-3-en-1-yloxy)benzoate (19b). Obtained following the general procedure A from 18a. Colorless oil (6.0 g, 88%). 1H NMR (500 MHz, CDCl3) δ 7.77 (dd, J = 7.5, 1.5 Hz, 1H), 7.47−7.38 (m, 1H), 7.00−6.91 (m, 2H), 6.00−5.88 (m, 1H), 5.22−5.07 (m, 2H), 4.08 (t, J = 6.5 Hz, 2H), 3.88 (s, 3H), 2.62−2.54 (m, 2H) ppm. ESI-MS: m/z = 207 [M + H]+. Methyl 2-(Pent-4-en-1-yloxy)benzoate (19c). Obtained following the general procedure A from 18a. Colorless oil (5.8 g, 80%). 1 H NMR (500 MHz, CDCl3) δ 7.78 (dd, J = 7.5, 1.5 Hz, 1H), 7.44 (ddd, J = 8.0, 7.5, 1.5 Hz, 1H), 6.99−6.94 (m, 2H), 5.92−5.80 (m, 1H), 5.09−5.04 (m, 1H), 5.02−4.98 (m, 1H), 4.05 (t, J = 6.5 Hz, 2H), 3.89 (s, 3H), 2.34−2.25 (m, 2H), 1.98−1.88 (m, 2H) ppm. ESIMS: m/z = 221 [M + H]+. Methyl 2-(Hex-5-en-1-yloxy)benzoate (19d). Obtained following the general procedure A from 18a. Colorless oil (6.3 g, 81%). 1H NMR (500 MHz, CDCl3) δ 7.78 (d, J = 8.0 Hz, 1H), 7.47−7.41 (m, 1H), 6.99−6.93 (m, 2H), 5.88−5.78 (m, 1H), 5.08−4.94 (m, 2H), 4.04 (t, J = 6.5 Hz, 2H), 3.88 (s, 3H), 2.17−2.10 (m, 2H), 1.90−1.81 (m, 2H), 1.65−1.57 (m, 2H) ppm. ESI-MS: m/z = 235 [M + H]+. Methyl 2-(Hept-6-en-1-yloxy)benzoate (19e). Obtained following the general procedure A from 18a. Colorless oil (6.1 g, 75%). 1 H NMR (500 MHz, CDCl3) δ 7.78 (d, J = 8.0 Hz, 1H), 7.46−7.41 (m, 1H), 6.98−6.93 (m, 2H), 5.89−5.76 (m, 1H), 5.04−4.90 (m, 2H), 4.03 (t, J = 6.5 Hz, 2H), 3.89 (s, 3H), 2.13−2.06 (m, 2H), 1.89−1.80 (m, 2H), 1.56−1.42 (m, 4H) ppm. 13C NMR (100 MHz, CDCl3) δ 167.11, 158.69, 138.92, 133.37, 131.67, 120.71, 120.14, 114.53, 113.37, 68.98, 51.96, 33.79, 29.15, 28.72, 25.59. ESI-MS: m/z = 249 [M + H]+. Methyl 2-(Oct-7-en-1-yloxy)benzoate (19f). Obtained following the general procedure A from 18a. Colorless oil (6.1 g, 70%). 1H NMR (500 MHz, CDCl3) δ 7.78 (d, J = 8.0 Hz, 1H), 7.46−7.40 (m, 1H), 6.99−6.92 (m, 2H), 5.87−5.75 (m, 1H), 5.05−4.90 (m, 2H), 4.03 (t, J = 6.5 Hz, 2H), 3.89 (s, 3H), 2.10−2.01 (m, 2H), 1.88−1.79 (m, 2H), 1.55−1.46 (m, 2H), 1.46−1.34 (m, 4H) ppm. 13C NMR (100 MHz, CDCl3) δ 167.15, 158.75, 139.14, 133.40, 131.70, 120.71, 120.15, 114.41, 113.41, 69.07, 51.99, 33.82, 29.27, 28.98, 28.93, 25.94. ESI-MS: m/z = 263 [M + H]+. Methyl 3-Fluoro-2-(pent-4-en-1-yloxy)benzoate (19g). Obtained following the general procedure A from commercial methyl 3fluoro-2-hydroxybenzoate 18b. Colorless oil (6.2 g, 80%). 1H NMR (500 MHz, CDCl3) δ 7.53 (dt, J = 8.0, 1.5 Hz, 1H), 7.23 (ddd, J = 11.0, 8.0, 1.5 Hz, 1H), 7.05 (td, J = 8.0, 4.5 Hz, 1H), 5.92−5.81 (m, 1H), 5.10−4.96 (m, 2H), 4.12 (td, J = 6.5, 1.0 Hz, 2H), 3.91 (s, 3H), 2.30−2.22 (m, 2H), 1.95−1.86 (m, 2H). 13C NMR (100 MHz, CDCl3) δ 166.11 (d, JC−F = 3 Hz), 156.42 (d, 1JC−F = 246 Hz), 147.10 (d, 2JC−F = 13 Hz), 138.09, 126.34 (d, 4JC−F = 3 Hz), 123.48 (d, 3JC−F = 8 Hz), 120.40 (d, 2JC−F = 19 Hz), 115.15, 74.64 (d, JC−F = 4 Hz), 52.41, 30.08, 29.45. ESI-MS: m/z = 239 [M + H]+. Methyl 3-Methoxy-2-(pent-4-en-1-yloxy)benzoate (19h). Obtained following the general procedure A from commercial methyl 2-hydroxy-3-methoxybenzoate 18c. Colorless oil (7.4 g, 90%). 1H NMR (500 MHz, CDCl3) δ 7.31 (dd, J = 7.5, 2.0 Hz, 1H), 7.09−7.02 (m, 2H), 5.93−5.84 (m, 1H), 5.09−5.03 (m, 1H), 5.00−4.96 (m, 1H), 4.04 (t, J = 6.5 Hz, 2H), 3.89 (s, 3H), 3.86 (s, 3H), 2.29−2.21 (m, 2H), 1.93−1.84 (m, 2H) ppm. ESI-MS: m/z = 251 [M + H]+. Methyl 4-Fluoro-2-(pent-4-en-1-yloxy)benzoate (19i). Obtained following the general procedure A from methyl 4-fluoro-2hydroxybenzoate 18d. Colorless oil (6.4 g, 82%). 1H NMR (500 MHz, CDCl3) δ 7.86−7.81 (m, 1H), 6.68−6.63 (m, 2H), 5.90−5.80 (m, 1H), 5.10−5.04 (m, 1H), 5.03−4.99 (m, 1H), 4.02 (t, J = 6.5 Hz,

2H), 3.88 (s, 3H), 2.32−2.26 (m, 2H), 1.98−1.91 (m, 2H) ppm. ESIMS: m/z = 239 [M + H]+. Methyl 4-Methoxy-2-(pent-4-en-1-yloxy)benzoate (19j). Obtained following the general procedure A from methyl 2hydroxy-4-methoxybenzoate 18e. Colorless oil (6.9 g, 84%). 1H NMR (500 MHz, CDCl3) δ 7.84 (d, J = 8.5 Hz, 1H), 6.49 (dd, J = 8.5, 2.5 Hz, 1H), 6.46 (d, J = 2.5 Hz, 1H), 5.91−5.81 (m, 1H), 5.09− 5.03 (m, 1H), 5.02−4.97 (m, 1H), 4.02 (t, J = 6.5 Hz, 2H), 3.85 (s, 3H), 3.84 (s, 3H), 2.33−2.26 (m, 2H), 1.98−1.91 (m, 2H) ppm. ESIMS: m/z = 251 [M + H]+. Methyl 4-Methyl-2-(pent-4-en-1-yloxy)benzoate (19k). Obtained following the general procedure A from methyl 2-hydroxy-4methylbenzoate 18f. Colorless oil (6.1 g, 79%). 1H NMR (500 MHz, CDCl3) δ 7.71 (d, J = 7.5 Hz, 1H), 6.80−6.74 (m, 2H), 5.92−5.82 (m, 1H), 5.10−5.04 (m, 1H), 5.02−4.97 (m, 1H), 4.03 (t, J = 6.4 Hz, 2H), 3.87 (s, 3H), 2.36 (s, 3H), 2.32−2.26 (m, 2H), 1.97−1.90 (m, 2H) ppm. ESI-MS: m/z = 235 [M + H]+ Methyl 4-Chloro-2-(pent-4-en-1-yloxy)benzoate (19l). Obtained following the general procedure A from methyl 4-chloro-2hydroxybenzoate 18g. Colorless oil (7.1 g, 85%). 1H NMR (500 MHz, CDCl3) δ 7.77−7.72 (m, 1H), 6.97−6.93 (m, 2H), 5.90−5.80 (m, 1H), 5.10−4.99 (m, 2H), 4.03 (t, J = 6.5 Hz, 2H), 3.88 (s, 3H), 2.32−2.25 (m, 2H), 1.98−1.90 (m, 2H) ppm. ESI-MS: m/z = 255 [M + H]+. Methyl 5-Chloro-2-(pent-4-en-1-yloxy)benzoate (19m). Obtained following the general procedure A from methyl 5-chloro-2hydroxybenzoate 18h. Colorless oil (6.9 g, 82%). 1H NMR (500 MHz, CDCl3) δ 7.75 (d, J = 3.0 Hz, 1H), 7.38 (dd, J = 9.0, 3.0 Hz, 1H), 6.89 (d, J = 9.0 Hz, 1H), 5.90−5.79 (m, 1H), 5.09−5.03 (m, 1H), 5.02−4.97 (m, 1H), 4.02 (t, J = 6.5 Hz, 2H), 3.89 (s, 3H), 2.31−2.24 (m, 2H), 1.96−1.87 (m, 2H) ppm. 13C NMR (100 MHz, CDCl3) δ 165.78, 157.33, 137.76, 133.11, 131.43, 125.30, 121.94, 115.48, 114.84, 68.68, 52.27, 30.07, 28.41. ESI-MS: m/z = 255 [M + H]+. Methyl 3-(Pent-4-en-1-yloxy)picolinate (19n). Obtained following the general procedure A from methyl 3-hydroxypicolinate 18i. Colorless oil (6.4 g, 89%). 1H NMR (500 MHz, CDCl3) δ 8.25 (dd, J = 4.5, 1.5 Hz, 1H), 7.38 (dd, J = 8.5, 4.5 Hz, 1H), 7.33 (dd, J = 8.5, 1.5 Hz, 1H), 5.89−5.78 (m, 1H), 5.09−5.03 (m, 1H), 5.02−4.97 (m, 1H), 4.06 (t, J = 6.5 Hz, 2H), 3.96 (s, 3H), 2.31−2.23 (m, 2H), 1.97−1.89 (m, 2H) ppm. 13C NMR (100 MHz, CDCl3) δ 165.30, 154.98, 140.97, 139.28, 137.48, 126.98, 121.12, 115.56, 68.30, 52.54, 29.90, 28.14. ESI-MS: m/z = 222 [M + H]+. Methyl 3-(Hept-6-en-1-yloxy)benzoate (19o). Obtained following the general procedure A from commercial methyl 3hydroxybenzoate 18j. Colorless oil (6.7 g, 82%). 1H NMR (500 MHz, CDCl3) δ 7.62 (d, J = 7.5 Hz, 1H), 7.57−7.52 (m, 1H), 7.36− 7.30 (m, 1H), 7.09 (dd, J = 8.0, 2.0 Hz, 1H), 5.87−5.76 (m, 1H), 5.04−4.98 (m, 1H), 4.98−4.93 (m, 1H), 4.00 (t, J = 6.5 Hz, 2H), 3.91 (s, 3H), 2.14−2.06 (m, 2H), 1.83−1.77 (m, 2H), 1.55−1.40 (m, 4H) ppm. ESI-MS: m/z = 249 [M + H]+. Ethyl or Pent-4-en-1-yl 1-Methyl-5-(pent-4-en-1-yloxy)-1Hpyrazole-4-carboxylate (19p). A solution of pent-4-en-1-ol (1.45 mL, 14.0 mmol) in THF (24.0 mL) was cooled to −8 °C and treated with 1.0 M sodium hexamethyldisilazane in THF (14.0 mL, 14.0 mmol). The reaction mixture was stirred at the reduced temperature for 5 min before the ice bath was removed. Stirring was continued for an additional 25 min before being treated with a solution of ethyl 5bromo-1-methyl-1H-pyrazole-4-carboxylate (18k, 1.1 g, 4.7 mmol) in THF (24 mL). The reaction mixture was stirred for 1 h at room temperature before being quenched with 50 mL of satd NH4Cl. The crude product was extracted with DCM (60 mL × 3), dried with Na2SO4, and concentrated under reduced pressure. This material was chromatographed on silica, eluting with 10−20% EtOAc in PE to afford 0.51 g product mixed with ethyl and pent-4-en-1-yl esters. Pent-4-en-1-yl 2-(pent-4-en-1-yloxy)nicotinate (19q). Compound 19q was synthesized following the general procedure provided for the synthesis of 19p. Light-yellow oil (0.52 g, 40%). 1H NMR (500 MHz, CDCl3) δ 8.28 (dd, J = 5.0, 2.0 Hz, 1H), 8.14 (dd, J = 7.5, 2.0 Hz, 1H), 6.92 (dd, J = 7.5, 5.0 Hz, 1H), 5.91−5.78 (m, 2H), 9189



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Ethyl 5-Allyl-1-methyl-1H-pyrazole-4-carboxylate (19x). Commercial ethyl 5-bromo-1-methyl-1H-pyrazole-4-carboxylate 18k (1.2 g, 5.3 mmol), allyltributyltin (1.8 mL, 5.8 mmol), Pd2(dba)3 (240 mg, 5 mol %), PtBu3 (0.27 mL, 18 mol %), and CsF (1.5 g, 26 mmol) was charged in a three-necked flask under N2. To this mixture was added dry THF (25 mL), and the reaction mixture was heated to reflux for 5 h. The reaction mixture was filtered off, the solvent removed in a vacuum, and the crude product purified by flash chromatography (30% EtOAc/PE) to give 19x as a viscous liquid (0.82 g, 80%). 1H NMR (500 MHz, CDCl3) δ 7.85 (s, 1H), 5.94− 5.80 (m, 1H), 5.16−5.06 (m, 1H), 5.02−4.90 (m, 1H), 4.28 (q, J = 7.0 Hz, 2H), 3.85−3.73 (m, 5H), 1.34 (t, J = 7.0 Hz, 3H) ppm. 13C NMR (100 MHz, CDCl3) δ 163.68, 144.02, 140.84, 132.76, 116.94, 112.03, 59.93, 36.60, 28.83, 14.48. ESI-MS: m/z = 195 [M + H]+. Methyl 3-(Pent-4-en-1-yloxy)thiophene-2-carboxylate (19y). Obtained following the general procedure A from commercially available methyl 3-hydroxythiophene-2-carboxylate 18q. White solid (6.2 g, 83%); mp = 69−70 °C. 1H NMR (500 MHz, CDCl3) δ 7.39 (d, J = 5.5 Hz, 1H), 6.83 (d, J = 5.5 Hz, 1H), 5.91−5.80 (m, 1H), 5.09−5.03 (m, 1H), 5.02−4.98 (m, 1H), 4.14 (t, J = 6.5 Hz, 2H), 3.84 (s, 3H), 2.30−2.24 (m, 2H), 1.97−1.90 (m, 2H) ppm. 13C NMR (100 MHz, CDCl3) δ 162.29, 161.43, 137.66, 130.57, 117.10, 115.43, 71.27, 51.61, 29.91, 28.54. ESI-MS: m/z = 227[M + H]+. Ethyl 2-Methyl-4-(pent-4-en-1-yloxy)thiazole-5-carboxylate (19z). Obtained following the general procedure A from commercial ethyl 4-hydroxy-2-methylthiazole-5-carboxylate 18r. Viscous liquid (6.6 g, 78%). 1H NMR (500 MHz, CDCl3) δ 5.92−5.78 (m, 1H), 5.09−5.03 (m, 1H), 5.01−4.96 (m, 1H), 4.45 (t, J = 6.5 Hz, 2H), 4.28 (q, J = 7.0 Hz, 2H), 2.61 (s, 3H), 2.28−2.21 (m, 2H), 1.96−1.87 (m, 2H), 1.33 (t, J = 7.0 Hz, 3H) ppm. 13C NMR (100 MHz, CDCl3) δ 167.82, 165.13, 161.64, 137.92, 115.22, 100.65, 70.46, 60.70, 30.02, 28.69, 20.28, 14.52. ESI-MS: m/z = 256[M + H]+. 2-(Allyloxy)benzoic Acid (20a). Obtained following the general procedure B from 19a. White solid (5.3 g, 96%); mp = 63−64 °C. 1H NMR (500 MHz, CDCl3) δ 8.80 (d, J = 6.5 Hz, 1H), 8.19 (d, J = 8.0 Hz, 1H), 7.48−7.40 (m, 2H), 7.39−7.29 (m, 5H), 7.11−7.04 (m, 1H), 6.97 (d, J = 8.3 Hz, 1H), 6.24−6.11 (m, 1H), 5.80−5.70 (m, 1H), 5.48−5.30 (m, 2H), 5.27−5.08 (m, 4H), 4.87−4.77 (m, 2H), 4.75−4.67 (m, 2H), 3.97−3.87 (m, 4H), 3.71−3.65 (m, 1H), 3.56− 3.49 (m, 1H), 3.38 (s, 3H) ppm. ESI-MS: m/z = 179 [M + H]+. 2-(But-3-en-1-yloxy)benzoic Acid (20b). Obtained following the general procedure B from 19b. Colorless oil (5.5 g, 98%). ESIMS: m/z = 193 [M + H]+. 2-(Pent-4-en-1-yloxy)benzoic Acid (20c). Obtained following the general procedure B from 19c. Colorless oil (4.9 g, 91%). 1H NMR (500 MHz, CDCl3) δ 8.15 (dd, J = 8.0, 2.0 Hz, 1H), 7.54 (ddd, J = 8.5, 7.5, 2.0 Hz, 1H), 7.11 (m, 1H), 7.03 (d, J = 8.5 Hz, 1H), 5.88−5.76 (m, 1H), 5.13−5.00 (m, 2H), 4.25 (t, J = 6.5 Hz, 2H), 2.30−2.22 (m, 2H), 2.06−1.97 (m, 2H) ppm. 13C NMR (100 MHz, CDCl3) δ 166.04, 157.75, 136.69, 135.09, 133.74, 122.13, 117.89, 116.26, 112.81, 69.49, 29.95, 28.11. ESI-MS: m/z = 207 [M + H]+. 2-(Hex-5-en-1-yloxy)benzoic Acid (20d). Obtained following the general procedure B from 19d. Colorless oil (5.4 g, 91%). 1H NMR (500 MHz, CDCl3) δ 8.17 (d, J = 8.0 Hz, 1H), 7.54 (ddd, J = 8.5, 7.5, 2.0 Hz, 1H), 7.16−7.09 (m, 1H), 7.04 (d, J = 8.5 Hz, 1H), 5.86−5.74 (m, 1H), 5.09−4.98 (m, 2H), 4.25 (t, J = 6.5 Hz, 2H), 2.20−2.11 (m, 2H), 1.97−1.89 (m, 2H), 1.65−1.54 (m, 2H) ppm. 13 C NMR (100 MHz, CDCl3) δ 165.62, 157.68, 137.80, 135.10, 133.82, 122.22, 117.83, 115.50, 112.73, 70.18, 33.21, 28.38, 25.14. ESI-MS: m/z = 221 [M + H]+. 2-(Hept-6-en-1-yloxy)benzoic Acid (20e). Obtained following the general procedure B from 19e. Colorless oil (5.4 g, 93%). 1H NMR (500 MHz, CDCl3) δ 8.14 (dd, J = 8.0, 2.0 Hz, 1H), 7.53 (ddd, J = 8.5, 7.5, 2.0 Hz, 1H), 7.13−7.06 (m, 1H), 7.03 (d, J = 8.5 Hz, 1H), 5.84−5.72 (m, 1H), 5.05−4.92 (m, 2H), 4.23 (t, J = 6.5 Hz, 2H), 2.13−2.03 (m, 2H), 1.96−1.86 (m, 2H), 1.56−1.43 (m, 4H) ppm. 13C NMR (100 MHz, CDCl3) δ 165.88, 157.76, 138.38, 135.11, 133.79, 122.15, 117.83, 114.92, 112.74, 70.25, 33.52, 28.87, 28.46, 25.37. ESI-MS: m/z = 235 [M + H]+.

5.09−5.07 (m, 1H), 5.05−5.03 (m, 1H), 5.03−4.96 (m, 2H), 4.42 (t, J = 6.5 Hz, 2H), 4.32 (t, J = 6.5 Hz, 2H), 2.29−2.19 (m, 4H), 1.95− 1.82 (m, 4H) ppm. ESI-MS: m/z = 276 [M + H]+. Methyl Pent-4-enoyl-L-prolinate (19r). Commercially available pent-4-enoic acid (1.0 g, 10.0 mmol) was dissolved in DCM (30 mL) and cooled to 0 °C. It was sequentially treated with EDCI (3.5 g, 18 mmol) and HOBt (1.6 g, 12 mmol). After stirring at room temperature for 30 min, methyl L-prolinate hydrochloride (18m, 2.0 g, 12 mmol) and DIPEA (5.0 mmol, 30 mmol) were added to the reaction mixture and stirred for 30 min, raising the temperature to room temperature, and stirred for 6 h. The residue was washed with saturated NaHCO3 (50 mL × 2) and brine (30 mL × 2), dried over Na2SO4, filtered, and concentrated in vacuo. Chromatographic purification of the residue provided 19r (1.9 g, 92%) as a colorless oil. 1H NMR (500 MHz, CDCl3) δ 5.93−5.80 (m, 1H), 5.09−4.96 (m, 2H), 4.50 (dd, J = 8.5, 3.5 Hz, 1H), 3.73 (s, 3H), 3.68−3.62 (m, 1H), 3.54−3.46 (m, 1H), 2.47−2.36 (m, 4H), 2.21−2.14 (m, 1H), 2.11−2.03 (m, 1H), 2.03−1.94 (m, 2H) ppm. 13C NMR (100 MHz, CDCl3) δ 172.97, 171.26, 137.50, 115.17, 58.66, 52.18, 47.03, 33.77, 29.26, 28.69, 24.82. ESI-MS: m/z = 212 [M + H]+ Methyl 1-(Pent-4-en-1-yl)-1H-pyrrole-2-carboxylate (19s). A mixture of commercial methyl 1H-pyrrole-2-carboxylate (18n, 1.0 g, 8.0 mmol), potassium carbonate (2.2 g, 16 mmol), KI (0.13 g, 0.80 mmol), and 5-bromopent-1-ene (1.1 mL, 9.6 mmol) in 10 mL of DMF was heated to 80 °C for 3 h. After cooling to room temperature, the solution was diluted with EtOAc (30 mL) and washed with water (20 mL × 2), brine (20 mL × 2). The organic layer was dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was then purified by flash chromatography on silica gel to give 19s (0.46 g, 30%) as a colorless oil. 1H NMR (500 MHz, CDCl3) δ 6.95 (dd, J = 4.0, 2.0 Hz, 1H), 6.84−6.82 (m, 1H), 6.12 (dd, J = 4.0, 2.5 Hz, 1H), 5.86−5.76 (m, 1H), 5.08−4.98 (m, 2H), 4.34−4.28 (m, 2H), 3.81 (s, 3H), 2.10−2.03 (m, 2H), 1.91−1.83 (m, 2H) ppm. ESIMS: m/z = 194 [M + H]+. Ethyl 1-(Pent-4-en-1-yl)-1H-imidazole-2-carboxylate (19t). Obtained following the general procedure A from commercial ethyl 1H-imidazole-2-carboxylate 18o and 5-bromopent-1-ene. Colorless oil (5.6 g, 81%). 1H NMR (500 MHz, CDCl3) δ 7.15 (d, J = 1.0 Hz, 1H), 7.06 (d, J = 1.0 Hz, 1H), 5.85−5.73 (m, 1H), 5.08−4.99 (m, 2H), 4.44−4.35 (m, 4H), 2.14−2.05 (m, 2H), 1.96−1.86 (m, 2H), 1.43 (t, J = 7.0 Hz, 3H) ppm. 13C NMR (100 MHz, CDCl3) δ 159.10, 136.99, 136.21, 129.43, 125.25, 115.83, 61.42, 47.94, 30.53, 30.24, 14.30. ESI-MS: m/z = 209 [M + H]+. Ethyl 1-(Hex-5-en-1-yl)-1H-imidazole-2-carboxylate (19u). Obtained following the general procedure A from 18o and 6bromohex-1-ene. Viscous liquid (6.2 g, 84%). 1H NMR (500 MHz, CDCl3) δ 7.14 (d, J = 1.0 Hz, 1H), 7.06 (d, J = 1.0 Hz, 1H), 5.80− 5.70 (m, 1H), 5.03−4.93 (m, 2H), 4.43−4.36 (m, 4H), 2.11−2.04 (m, 2H), 1.84−1.77 (m, 2H), 1.45−1.38 (m, 5H) ppm. 13C NMR (100 MHz, CDCl3) δ 159.16, 138.01, 136.23, 129.47, 125.14, 115.12, 61.40, 48.41, 33.17, 30.69, 25.78, 14.32. ESI-MS: m/z = 223 [M + H]+. Methyl 1-(Pent-4-en-1-yl)-1H-pyrazole-5-carboxylate (19v). Obtained following the general procedure A from commercial methyl 1H-pyrazole-5-carboxylate 18p and 5-bromopent-1-ene. Viscous liquid (5.7 g, 89%). 1H NMR (500 MHz, CDCl3) δ 7.48 (d, J = 2.0 Hz, 1H), 6.83 (d, J = 2.0 Hz, 1H), 5.86−5.76 (m, 1H), 5.08−5.01 (m, 1H), 5.00−4.96 (m, 1H), 4.62−4.54 (m, 2H), 3.88 (s, 3H), 2.12−2.04 (m, 2H), 1.99−1.90 (m, 2H) ppm. ESI-MS: m/z = 195 [M + H]+. Methyl 1-(Hex-5-en-1-yl)-1H-pyrazole-5-carboxylate (19w). Obtained following the general procedure A from 18p and 6bromohex-1-ene. Viscous liquid (6.2 g, 90%). 1H NMR (500 MHz, CDCl3) δ 7.47 (d, J = 2.0 Hz, 1H), 6.82 (d, J = 2.0 Hz, 1H), 5.82− 5.72 (m, 1H), 5.02−4.90 (m, 2H), 4.57 (t, J = 7.5 Hz, 2H), 3.87 (s, 3H), 2.11−2.04 (m, 2H), 1.89−1.80 (m, 2H), 1.44−1.36 (m, 2H) ppm. 13C NMR (100 MHz, CDCl3) δ 160.32, 138.44, 137.95, 131.70, 114.86, 111.43, 51.93, 51.74, 33.34, 30.15, 25.90. ESI-MS: m/z = 209 [M + H]+. 9190



Article

3-(Pent-4-en-1-yloxy)picolinic Acid (20n). Obtained following the general procedure B from 19n. White solid (4.9 g, 82%); mp = 115−116 °C. 1H NMR (500 MHz, CDCl3) δ 8.20 (dd, J = 4.5, 1.0 Hz, 1H), 7.53 (dd, J = 8.5, 4.5 Hz, 1H), 7.45 (dd, J = 8.5, 1.0 Hz, 1H), 5.93−5.76 (m, 1H), 5.12−4.95 (m, 2H), 4.15 (t, J = 6.5 Hz, 2H), 2.39−2.24 (m, 2H), 2.07−1.92 (m, 2H) ppm. 13C NMR (100 MHz, CDCl3) δ 162.40, 156.93, 139.59, 137.40, 134.64, 129.29, 122.80, 115.70, 68.77, 29.87, 28.04. ESI-MS: m/z = 208 [M + H]+. 3-(Hept-6-en-1-yloxy)benzoic Acid (20o). Obtained following the general procedure B from 19o. White solid (5.8 g, 92%); mp = 69−70 °C. 1H NMR (500 MHz, CDCl3) δ 7.74−7.68 (m, 1H), 7.62 (dd, J = 2.5, 1.5 Hz, 1H), 7.37 (t, J = 8.0 Hz, 1H), 7.15 (ddd, J = 8.5, 2.5, 1.0 Hz, 1H), 5.88−5.77 (m, 1H), 5.06−4.93 (m, 2H), 4.02 (t, J = 6.5 Hz, 2H), 2.14−2.06 (m, 2H), 1.87−1.78 (m, 2H), 1.56−1.44 (m, 4H) ppm. 13C NMR (100 MHz, CDCl3) δ 172.43, 159.31, 138.89, 130.68, 129.62, 122.65, 121.08, 115.27, 114.64, 68.33, 33.80, 29.17, 28.77, 25.66. ESI-MS: m/z = 235 [M + H]+. 1-Methyl-5-(pent-4-en-1-yloxy)-1H-pyrazole-4-carboxylic Acid (20p). Obtained following the general procedure B from 19p. White solid (0.36 g in two steps, 36%); mp = 67−68 °C. ESI-MS: m/ z = 211 [M + H]+. 2-(Pent-4-en-1-yloxy)nicotinic Acid (20q). Obtained following the general procedure B from 19q. White solid (0.33 g, 85%); mp = 85−86 °C. 1H NMR (500 MHz, DMSO) δ 14.66 (s, 1H), 8.37 (dd, J = 7.0, 2.0 Hz, 1H), 8.26 (dd, J = 6.5, 2.0 Hz, 1H), 6.73 (dd, J = 7.0, 6.5 Hz, 1H), 5.92−5.72 (m, 1H), 5.15−4.89 (m, 2H), 4.10 (t, J = 6.5 Hz, 2H), 2.15−2.00 (m, 2H), 1.89−1.75 (m, 2H) ppm. 13C NMR (100 MHz, DMSO) δ 164.78, 163.63, 145.11, 144.99, 137.28, 116.44, 115.42, 108.41, 49.59, 29.95, 27.37. ESI-MS: m/z = 208 [M + H]+. Pent-4-enoyl-L-proline (20r). To a solution of methyl pent-4enoyl-L-prolinate (1.9 g, 9.0 mmol) in acetone (30 mL) was added aqueous LiOH 0.3 M (1.5 equiv) dropwise. The solution was stirred at room temperature for 10 h. The reaction mixture was then acidified to pH 2 with 3 N HCl and extracted with EtOAc, washed with brine, dried over Na2SO4, filtered, and concentrated to yield 20r. Colorless oil (1.5 g, 82%). 1H NMR (500 MHz, DMSO) δ 5.89−5.78 (m, 1H), 5.08−4.90 (m, 2H), 4.21 (dd, J = 9.0, 4.0 Hz, 1H), 3.56−3.45 (m, 2H), 2.38−2.29 (m, 2H), 2.26−2.19 (m, 2H), 2.14−2.07 (m, 1H), 1.93−1.79 (m, 3H) ppm. 13C NMR (100 MHz, CDCl3) δ 173.77, 173.64, 137.01, 115.68, 59.56, 47.72, 33.85, 28.68, 28.29, 24.80. ESIMS: m/z = 198 [M + H]+. 1-(Pent-4-en-1-yl)-1H-pyrrole-2-carboxylic Acid (20s). Obtained following the general procedure B from 19s. Colorless oil (0.39 g, 92%). ESI-MS: m/z = 180 [M + H]+. 1-(Pent-4-en-1-yl)-1H-imidazole-2-carboxylic Acid (20t). Obtained following the general procedure B from 19t. Colorless oil (3.5 g, 72%). ESI-MS: m/z = 181 [M + H]+. 1-(Hex-5-en-1-yl)-1H-imidazole-2-carboxylic Acid (20u). Obtained following the general procedure B from 19u. Colorless oil (4.0 g, 75%). ESI-MS: m/z = 195 [M + H]+. 1-(Pent-4-en-1-yl)-1H-pyrazole-5-carboxylic Acid (20v). Obtained following the general procedure B from 19v. White solid (4.7 g, 95%); mp = 60−61 °C. 1H NMR (500 MHz, CDCl3) δ 7.59 (d, J = 2.0 Hz, 1H), 6.97 (d, J = 2.0 Hz, 1H), 5.89−5.72 (m, 1H), 5.09−4.92 (m, 2H), 4.64 (t, J = 7.5 Hz, 2H), 2.16−2.04 (m, 2H), 2.03−1.89 (m, 2H) ppm. 13C NMR (100 MHz, CDCl3) δ 163.18, 138.00, 137.36, 132.05, 115.43, 112.61, 51.45, 30.70, 29.84. ESI-MS: m/z = 181 [M + H]+. 1-(Hex-5-en-1-yl)-1H-pyrazole-5-carboxylic Acid (20w). Obtained following the general procedure B from 19w. White solid (5.1 g, 95%), mp = 99−100 °C. 1H NMR (500 MHz, CDCl3) δ 11.90 (s, 1H), 7.60 (d, J = 2.0 Hz, 1H), 6.95 (d, J = 2.0 Hz, 1H), 5.82−5.68 (m, 1H), 5.04−4.85 (m, 2H), 4.65 (t, J = 7.5 Hz, 2H), 2.15−1.98 (m, 2H), 1.95−1.81 (m, 2H), 1.51−1.33 (m, 2H) ppm. 13C NMR (100 MHz, CDCl3) δ 163.18, 138.00, 137.36, 132.05, 115.43, 112.61, 51.45, 30.70, 29.84. ESI-MS: m/z = 195 [M + H]+. 5-Allyl-1-methyl-1H-pyrazole-4-carboxylic Acid (20x). Obtained following the general procedure B from 19x. White solid (0.65 g, 95%), mp = 120−121 °C. 1H NMR (500 MHz, DMSO) δ 12.27 (s, 1H), 7.74 (s, 1H), 5.93−5.76 (m, 1H), 5.12−4.87 (m, 2H), 3.80−

2-(Oct-7-en-1-yloxy)benzoic Acid (20f). Obtained following the general procedure B from 19f. Colorless oil (5.2 g, 90%). 1H NMR (500 MHz, CDCl3) δ 8.15 (dd, J = 8.0, 2.0 Hz, 1H), 7.56−7.50 (m, 1H), 7.14−7.07 (m, 1H), 7.03 (d, J = 8.5 Hz, 1H), 5.85−5.71 (m, 1H), 5.04−4.88 (m, 2H), 4.23 (t, J = 6.5 Hz, 2H), 2.10−1.99 (m, 2H), 1.96−1.82 (m, 2H), 1.57−1.32 (m, 6H) ppm. 13C NMR (100 MHz, CDCl3) δ 166.12, 165.17, 157.81, 138.77, 135.17, 133.76, 122.12, 117.64, 114.58, 112.75, 70.29, 33.62, 28.95, 28.68, 25.77. ESIMS: m/z = 249 [M + H]+. 3-Fluoro-2-(pent-4-en-1-yloxy)benzoic Acid (20g). Obtained following the general procedure B from 19g. Colorless oil (5.2 g, 89%). 1H NMR (500 MHz, CDCl3) δ 7.85 (d, J = 8.0 Hz, 1H), 7.32 (ddd, J = 11.5, 8.0, 1.5 Hz, 1H), 7.14 (td, J = 8.0, 5.0 Hz, 1H), 5.88− 5.75 (m, 1H), 5.12−4.96 (m, 2H), 4.30 (t, J = 6.5 Hz, 2H), 2.29− 2.19 (m, 2H), 2.01−1.90 (m, 2H) ppm. 13C NMR (100 MHz, CDCl3) δ 166.66, 155.18 (d, 1JC−F = 247 Hz), 146.56 (d, 3JC−F = 12 Hz), 137.16, 127.86 (d, 4JC−F = 3 Hz), 124.12 (d, 3JC−F = 8 Hz), 124.00, 121.89 (d, 2JC−F = 20 Hz), 115.76, 75.53 (d, JC−F = 6 Hz), 29.76, 29.16. ESI-MS: m/z = 225 [M + H]+. 3-Methoxy-2-(pent-4-en-1-yloxy)benzoic Acid (20h). Obtained following the general procedure B from 19h. Colorless oil (6.1 g, 88%). 1H NMR (500 MHz, CDCl3) δ 7.73−7.67 (m, 1H), 7.21−7.11 (m, 2H), 5.87−5.76 (m, 1H), 5.11−4.97 (m, 2H), 4.26 (t, J = 6.5 Hz, 2H), 3.89 (s, 3H), 2.27−2.18 (m, 2H), 2.01−1.89 (m, 2H) ppm. 13C NMR (100 MHz, CDCl3) δ 165.61, 152.18, 147.46, 137.16, 124.87, 124.01, 122.58, 117.57, 115.77, 74.97, 56.30, 29.78, 29.17. ESI-MS: m/z = 237 [M + H]+. 4-Fluoro-2-(pent-4-en-1-yloxy)benzoic Acid (20i). Obtained following the general procedure B from 19i. Colorless oil (5.7 g, 95%). 1H NMR (500 MHz, CDCl3) δ 8.04 (t, J = 7.5 Hz, 1H), 6.75− 6.62 (m, 2H), 5.84−5.71 (m, 1H), 5.07−4.94 (m, 2H), 4.13 (t, J = 6.5 Hz, 2H), 2.28−2.15 (m, 2H), 2.00−1.88 (m, 2H) ppm. 13C NMR (100 MHz, CDCl3) δ 166.60 (d, 1JC−F = 254 Hz), 165.93, 159.60 (d, 3 JC−F = 10 Hz), 136.68, 135. 56 (d, 3JC−F = 10 Hz), 116.09, 114.33, 108.79 (d, 2JC−F = 22 Hz), 100.90 (d, 2JC−F = 27 Hz), 69.63, 29.81, 27.86. ESI-MS: m/z = 225 [M + H]+. 4-Methoxy-2-(pent-4-en-1-yloxy)benzoic Acid (20j). Obtained following the general procedure B from 19j. Colorless oil (6.2 g, 95%). 1H NMR (500 MHz, CDCl3) δ 8.13 (d, J = 9.0 Hz, 1H), 6.64 (dd, J = 9.0, 2.5 Hz, 1H), 6.51 (d, J = 2.5 Hz, 1H), 5.89− 5.76 (m, 1H), 5.14−5.01 (m, 2H), 4.22 (t, J = 6.5 Hz, 2H), 3.87 (s, 3H), 2.30−2.21 (m, 2H), 2.06−1.98 (m, 2H) ppm. 13C NMR (100 MHz, CDCl3) δ 165.38, 165.17, 159.03, 136.62, 135.66, 116.43, 110.74, 106.80, 99.66, 69.53, 55.86, 29.99, 28.08. ESI-MS: m/z = 237 [M + H]+. 4-Methyl-2-(pent-4-en-1-yloxy)benzoic Acid (20k). Obtained following the general procedure B from 19k. Colorless oil (5.3 g, 93%). 1H NMR (500 MHz, CDCl3) δ 8.04 (d, J = 8.0 Hz, 1H), 6.92 (dd, J = 8.0, 0.5 Hz, 1H), 6.83 (s, 1H), 5.89−5.76 (m, 1H), 5.14− 5.02 (m, 2H), 4.24 (t, J = 6.5 Hz, 2H), 2.40 (s, 3H), 2.31−2.21 (m, 2H), 2.07−1.95 (m, 2H) ppm. 13C NMR (100 MHz, CDCl3) δ 157.55, 146.46, 136.66, 133.67, 123.16, 116.31, 115.12, 113.33, 69.41, 29.96, 28.13, 22.03. ESI-MS: m/z = 221 [M + H]+. 4-Chloro-2-(pent-4-en-1-yloxy)benzoic Acid (20l). Obtained following the general procedure B from 19l. White solid (6.6 g, 98%); mp = 55−56 °C. 1H NMR (500 MHz, CDCl3) δ 8.06 (dd, J = 8.5, 2.5 Hz, 1H), 7.08 (dd, J = 8.5, 2.0 Hz, 1H), 7.02 (s, 1H), 5.89−5.75 (m, 1H), 5.14−5.00 (m, 2H), 4.21 (t, J = 6.5 Hz, 2H), 2.31−2.23 (m, 2H), 2.06−1.96 (m, 2H) ppm. 13C NMR (100 MHz, CDCl3) δ 165.71, 158.32, 140.93, 136.64, 134.73, 122.33, 116.64, 116.39, 113.55, 69.83, 29.92, 28.03. ESI-MS: m/z = 241 [M + H]+. 5-Chloro-2-(pent-4-en-1-yloxy)benzoic Acid (20m). Obtained following the general procedure B from 19m. Colorless oil (6.3 g, 96%). 1H NMR (500 MHz, CDCl3) δ 8.03 (d, J = 2.5 Hz, 1H), 7.45 (dd, J = 9.0, 2.5 Hz, 1H), 6.96 (d, J = 9.0 Hz, 1H), 5.86−5.73 (m, 1H), 5.11−4.99 (m, 2H), 4.19 (t, J = 6.5 Hz, 2H), 2.27−2.20 (m, 2H), 2.04−1.92 (m, 2H) ppm. 13C NMR (100 MHz, CDCl3) δ 165.17, 156.36, 136.62, 134.66, 133.03, 127.24, 119.28, 116.30, 114.39, 69.90, 29.87, 28.01. ESI-MS: m/z = 241 [M + H]+. 9191



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3.67 (m, 5H) ppm. 13C NMR (100 MHz, DMSO) δ 164.30, 143.59, 140.10, 133.40, 116.52, 111.62, 36.30, 27.81. ESI-MS: m/z = 167 [M + H]+. 3-(Pent-4-en-1-yloxy)thiophene-2-carboxylic Acid (20y). Obtained following the general procedure B from 19y. White solid (5.1 g, 88%); mp = 60−61 °C. 1H NMR (500 MHz, CDCl3) δ 7.51 (d, J = 5.5 Hz, 1H), 6.85 (d, J = 5.5 Hz, 1H), 5.89−5.76 (m, 1H), 5.11−4.99 (m, 2H), 4.21 (t, J = 6.5 Hz, 2H), 2.29−2.20 (m, 2H), 1.99−1.88 (m, 2H) ppm. 13C NMR (100 MHz, CDCl3) δ 164.66, 160.78, 137.15, 132.51, 116.43, 115.86, 113.00, 110.77, 105.55, 71.92, 29.83, 28.36. ESI-MS: m/z = 213 [M + H]+. 2-Methyl-4-(pent-4-en-1-yloxy)thiazole-5-carboxylic Acid (20z). Obtained following the general procedure B from 19z. White solid (5.5 g, 93%); mp = 79−80 °C. 1H NMR (500 MHz, CDCl3) δ 5.89−5.78 (m, 1H), 5.10−4.96 (m, 2H), 4.49 (t, J = 6.5 Hz, 2H), 2.62 (s, 3H), 2.25−2.17 (m, 2H), 1.95−1.86 (m, 2H) ppm. 13C NMR (100 MHz, CDCl3) δ 169.78, 165.95, 165.59, 137.59, 115.34, 100.38, 70.90, 29.87, 28.48, 20.37. ESI-MS: m/z = 228 [M + H]+. Benzyl O-Allyl-N-(N-(2-(allyloxy)benzoyl)-O-methyl-L-seryl)L-serinate (21a). Obtained following the general procedure B from 20a and 10a. Viscous liquid (two steps, 0.39 g, 78%). 1H NMR (500 MHz, CDCl3) δ 8.80 (d, J = 6.5 Hz, 1H), 8.19 (d, J = 8.0 Hz, 1H), 7.48−7.40 (m, 2H), 7.39−7.29 (m, 5H), 7.11−7.04 (m, 1H), 6.97 (d, J = 8.3 Hz, 1H), 6.24−6.11 (m, 1H), 5.80−5.70 (m, 1H), 5.48−5.30 (m, 2H), 5.27−5.08 (m, 4H), 4.87−4.77 (m, 2H), 4.75−4.67 (m, 2H), 3.97−3.87 (m, 4H), 3.71−3.65 (m, 1H), 3.56−3.49 (m, 1H), 3.38 (s, 3H) ppm. 13C NMR (100 MHz, CDCl3) δ 170.46, 169.90, 165.42, 157.02, 135.51, 134.12, 133.11, 132.52, 132.43, 128.61, 128.39, 128.17, 121.47, 118.97, 117.24, 112.87, 72.27, 71.83, 70.32, 69.71, 67.26, 59.08, 53.25, 53.17. ESI-MS: m/z = 497 [M + H]+. Benzyl O-Allyl-N-(N-(2-(but-3-en-1-yloxy)benzoyl)-O-methyl-L-seryl)-L-serinate (21b). Obtained following the general procedure B from 20b and 10a. Viscous liquid (two steps, 0.38 g, 77%). 1H NMR (500 MHz, CDCl3) δ 8.79 (d, J = 6.5 Hz, 1H), 8.20 (d, J = 7.5 Hz, 1H), 7.49−7.39 (m, 2H), 7.37−7.28 (m, 5H), 7.11− 7.04 (m, 1H), 6.97 (d, J = 8.5 Hz, 1H), 5.96−5.85 (m, 1H), 5.80− 5.68 (m, 1H), 5.27−5.07 (m, 6H), 4.90−4.82 (m, 1H), 4.82−4.77 (m, 1H), 4.18 (t, J = 6.5 Hz, 2H), 3.95−3.87 (m, 4H), 3.72−3.66 (m, 1H), 3.57−3.50 (m, 1H), 3.39 (s, 3H), 2.77−2.63 (m, 2H) ppm. 13C NMR (100 MHz, CDCl3) δ 170.42, 169.88, 165.38, 157.27, 135.47, 134.08, 134.02, 133.17, 132.43, 128.58, 128.36, 128.14, 121.22, 117.69, 117.23, 112.25, 72.23, 71.88, 69.66, 68.47, 67.22, 59.03, 53.16, 33.33. ESI-MS: m/z = 511 [M + H]+. Benzyl O-Allyl-N-(O-methyl-N-(2-(pent-4-en-1-yloxy)benzoyl)-L-seryl)-L-serinate (21c). Obtained following the general procedure B from 20c and 10a. Viscous liquid (two steps, 0.32 g, 61%). 1H NMR (500 MHz, CDCl3) δ 8.85 (d, J = 6.5 Hz, 1H), 8.21 (d, J = 8.0 Hz, 1H), 7.48−7.42 (m, 1H), 7.40 (d, J = 8.0 Hz, 1H), 7.36−7.29 (m, 5H), 7.09−7.03 (m, 1H), 6.96 (d, J = 8.0 Hz, 1H), 5.89−5.79 (m, 1H), 5.79−5.69 (m, 1H), 5.27−4.98 (m, 6H), 4.88− 4.82 (m, 1H), 4.81−4.77 (m, 1H), 4.16−4.09 (m, 2H), 3.97−3.87 (m, 4H), 3.70−3.65 (m, 1H), 3.55−3.49 (m, 1H), 3.38 (s, 3H), 2.30−2.22 (m, 2H), 2.11−1.97 (m, 2H) ppm. 13C NMR (100 MHz, CDCl3) δ 170.42, 169.88, 165.43, 157.48, 137.49, 135.50, 134.10, 133.17, 132.40, 128.59, 128.36, 128.15, 121.12, 117.20, 115.67, 112.28, 72.23, 71.89, 69.69, 68.59, 67.22, 59.08, 53.24, 53.15, 30.22, 28.20. ESI-MS: m/z = 525 [M + H]+. Benzyl O-Allyl-N-(N-(2-(hex-5-en-1-yloxy)benzoyl)-O-methyl-L-seryl)-L-serinate (21d). Obtained following the general procedure B from 20d and 10a. Viscous liquid (two steps, 0.35 g, 65%). 1H NMR (500 MHz, CDCl3) δ 8.85 (d, J = 6.5 Hz, 1H), 8.20 (dd, J = 8.0, 2.0 Hz, 1H), 7.44 (ddd, J = 8.5, 7.5, 2.0 Hz, 1H), 7.40 (d, J = 8.0 Hz, 1H), 7.37−7.28 (m, 5H), 7.09−7.03 (m, 1H), 6.96 (d, J = 8.0 Hz, 1H), 5.88−5.70 (m, 2H), 5.26−5.13 (m, 3H), 5.12−5.09 (m, 1H), 5.05−4.99 (m, 1H), 4.99−4.93 (m, 1H), 4.84 (td, J = 7.0, 4.0 Hz, 1H), 4.79 (dt, J = 8.0, 3.0 Hz, 1H), 4.17−4.08 (m, 2H), 3.97− 3.87 (m, 4H), 3.67 (dd, J = 9.5, 3.0 Hz, 1H), 3.53 (dd, J = 9.0, 6.5 Hz, 1H), 3.39 (s, 3H), 2.16−2.10 (m, 2H), 2.00−1.90 (m, 2H), 1.63− 1.54 (m, 2H) ppm. 13C NMR (100 MHz, CDCl3) δ 170.41, 169.86, 165.42, 157.49, 138.39, 135.48, 134.06, 133.14, 132.37, 128.57,

128.34, 128.13, 121.04, 117.19, 114.95, 112.25, 72.21, 71.88, 69.66, 69.14, 67.20, 59.07, 53.17, 53.12, 33.38, 28.49, 25.43. ESI-MS: m/z = 539 [M + H]+. Benzyl O-Allyl-N-(N-(2-(hept-6-en-1-yloxy)benzoyl)-Omethyl-L-seryl)-L-serinate (21e). Obtained following the general procedure B from 20e and 10a. Viscous liquid (two steps, 0.33 g, 60%). 1H NMR (500 MHz, CDCl3) δ 8.86 (d, J = 6.5 Hz, 1H), 8.20 (dd, J = 8.0, 2.0 Hz, 1H), 7.47−7.42 (m, 1H), 7.40 (d, J = 8.0 Hz, 1H), 7.38−7.28 (m, 5H), 7.09−7.02 (m, 1H), 6.96 (d, J = 8.0 Hz, 1H), 5.85−5.69 (m, 2H), 5.27−5.13 (m, 3H), 5.13−5.08 (m, 1H), 5.03−4.97 (m, 1H), 4.97−4.92 (m, 1H), 4.84 (td, J = 7.0, 4.0 Hz, 1H), 4.79 (dt, J = 8.0, 3.0 Hz, 1H), 4.15−4.09 (m, 2H), 3.96−3.87 (m, 4H), 3.67 (dd, J = 9.5, 3.5 Hz, 1H), 3.53 (dd, J = 9.0, 7.0 Hz, 1H), 3.39 (s, 3H), 2.11−2.05 (m, 2H), 1.99−1.91 (m, 2H), 1.53− 1.43 (m, 4H) ppm. 13C NMR (100 MHz, CDCl3) δ 170.49, 169.82, 167.19, 159.50, 138.88, 135.47, 135.28, 134.03, 129.66, 128.69, 128.52, 128.28, 119.02, 118.68, 117.45, 114.59, 113.16, 72.33, 71.76, 69.54, 68.26, 67.43, 59.18, 53.26, 52.51, 33.77, 29.17, 28.74, 25.63. ESI-MS: m/z = 553 [M + H]+. Benzyl O-Allyl-N-(O-methyl-N-(2-(oct-7-en-1-yloxy)benzoyl)-L-seryl)-L-serinate (21f). Obtained following the general procedure B from 20f and 10a. Viscous liquid (two steps, 0.31 g, 55%). 1H NMR (500 MHz, CDCl3) δ 8.85 (d, J = 7.0 Hz, 1H), 8.20 (dd, J = 8.0, 2.0 Hz, 1H), 7.44 (ddd, J = 8.0, 7.5, 2.0 Hz, 1H), 7.39 (d, J = 8.0 Hz, 1H), 7.36−7.30 (m, 6H), 7.08−7.03 (m, 1H), 6.96 (d, J = 8.0 Hz, 1H), 5.84−5.69 (m, 2H), 5.26−5.13 (m, 4H), 5.12−5.08 (m, 1H), 5.02−4.96 (m, 1H), 4.95−4.91 (m, 1H), 4.84 (td, J = 7.0, 4.0 Hz, 1H), 4.79 (dt, J = 8.0, 3.0 Hz, 1H), 3.95−3.88 (m, 4H), 3.67 (dd, J = 9.5, 3.5 Hz, 1H), 3.53 (dd, J = 9.0, 7.0 Hz, 1H), 3.38 (s, 3H), 2.07−2.02 (m, 2H), 1.98−1.90 (m, 2H), 1.51−1.44 (m, 2H), 1.44− 1.35 (m, 4H) ppm. ESI-MS: m/z = 567 [M + H]+. Benzyl O-Allyl-N-(N-(3-fluoro-2-(pent-4-en-1-yloxy)benzoyl)-O-methyl-L-seryl)-L-serinate (21g). Obtained following the general procedure B from 20g and 10a. Viscous liquid (two steps, 0.33 g, 61%). 1H NMR (500 MHz, CDCl3) δ 8.78 (d, J = 6.5 Hz, 1H), 7.90 (dt, J = 8.0, 1.5 Hz, 1H), 7.37−7.29 (m, 6H), 7.25−7.19 (m, 1H), 7.10 (td, J = 8.0, 4.5 Hz, 1H), 5.88−5.70 (m, 2H), 5.27− 5.10 (m, 4H), 5.08−5.02 (m, 1H), 5.01−4.97 (m, 1H), 4.84−4.76 (m, 2H), 4.28−4.16 (m, 2H), 3.97−3.88 (m, 4H), 3.68 (dd, J = 9.5, 3.0 Hz, 1H), 3.54 (dd, J = 9.0, 7.0 Hz, 1H), 3.38 (s, 3H), 2.26−2.20 (m, 2H), 2.01−1.94 (m, 2H) ppm. ESI-MS: m/z = 543 [M + H]+. Benzyl O-Allyl-N-(N-(3-methoxy-2-(pent-4-en-1-yloxy)benzoyl)-O-methyl-L-seryl)-L-serinate (21h). Obtained following the general procedure B from 20h and 10a. Viscous liquid (two steps, 0.36 g, 65%). 1H NMR (500 MHz, CDCl3) δ 8.93 (d, J = 7.0 Hz, 1H), 7.70 (dd, J = 8.0, 1.5 Hz, 1H), 7.39−7.28 (m, 6H), 7.13 (t, J = 8.0 Hz, 1H), 7.05 (dd, J = 8.0, 1.5 Hz, 1H), 5.90−5.80 (m, 1H), 5.78−5.69 (m, 1H), 5.26−5.13 (m, 3H), 5.13−5.08 (m, 1H), 5.08− 5.02 (m, 1H), 5.00−4.96 (m, 1H), 4.85−4.76 (m, 2H), 4.11−4.04 (m, 2H), 3.97−3.86 (m, 7H), 3.67 (dd, J = 9.5, 3.5 Hz, 1H), 3.54 (dd, J = 9.0, 6.5 Hz, 1H), 3.37 (s, 3H), 2.26−2.20 (m, 2H), 1.99−1.91 (m, 2H) ppm. 13C NMR (100 MHz, CDCl3) δ 170.19, 169.88, 165.54, 152.78, 147.30, 138.07, 138.01, 135.48, 134.08, 128.59, 128.36, 128.17, 124.10, 123.00, 117.26, 116.04, 114.98, 74.06, 72.25, 71.80, 69.64, 67.25, 59.07, 56.23, 53.25, 53.14, 30.00, 29.10. ESI-MS: m/z = 555 [M + H]+. Benzyl O-Allyl-N-(N-(4-fluoro-2-(pent-4-en-1-yloxy)benzoyl)-O-methyl-L-seryl)-L-serinate (21i). Obtained following the general procedure B from 20i and 10a. Viscous liquid (two steps, 0.40 g, 74%). 1H NMR (500 MHz, CDCl3) δ 8.72 (d, J = 6.5 Hz, 1H), 8.13 (d, J = 8.5 Hz, 1H), 7.37 (d, J = 8.5 Hz, 1H), 7.35−7.31 (m, 5H), 7.04 (dd, J = 8.5, 2.0 Hz, 1H), 6.95 (d, J = 2.0 Hz, 1H), 5.88−5.70 (m, 2H), 5.26−5.05 (m, 5H), 5.04−5.00 (m, 1H), 4.84− 4.76 (m, 2H), 4.11 (t, J = 6.5 Hz, 2H), 3.97−3.87 (m, 4H), 3.67 (dd, J = 9.5, 3.0 Hz, 1H), 3.52 (dd, J = 9.0, 7.0 Hz, 1H), 3.38 (s, 3H), 2.29−2.23 (m, 2H), 2.09−2.01 (m, 2H) ppm. ESI-MS: m/z = 543 [M + H]+. Benzyl O-Allyl-N-(N-(4-methoxy-2-(pent-4-en-1-yloxy)benzoyl)-O-methyl-L-seryl)-L-serinate (21j). Obtained following the general procedure B from 20j and 10a. Viscous liquid (two steps, 9192



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0.41 g, 76%). 1H NMR (500 MHz, CDCl3) δ 8.71 (d, J = 6.5 Hz, 1H), 8.16 (d, J = 8.5 Hz, 1H), 7.40 (d, J = 8.0 Hz, 1H), 7.37−7.28 (m, 5H), 6.58 (dd, J = 9.0, 2.0 Hz, 1H), 6.46 (d, J = 2.5 Hz, 1H), 5.88−5.69 (m, 2H), 5.26−5.03 (m, 5H), 5.02−4.98 (m, 1H), 4.83 (td, J = 7.0, 4.0 Hz, 1H), 4.78 (dt, J = 8.0, 3.0 Hz, 1H), 4.09 (t, J = 6.5 Hz, 2H), 3.96−3.87 (m, 4H), 3.84 (s, 3H), 3.67 (dd, J = 9.5, 3.0 Hz, 1H), 3.52 (dd, J = 9.5, 6.5 Hz, 1H), 3.37 (s, 3H), 2.30−2.20 (m, 2H), 2.09−1.98 (m, 2H) ppm. ESI-MS: m/z = 555 [M + H]+. Benzyl O-Allyl-N-(O-methyl-N-(4-methyl-2-(pent-4-en-1yloxy)benzoyl)-L-seryl)-L-serinate (21k). Obtained following the general procedure B from 20k and 10a. Viscous liquid (two steps, 0.44 g, 82%). 1H NMR (500 MHz, CDCl3) δ 8.80 (d, J = 7.0 Hz, 1H), 8.08 (d, J = 8.0 Hz, 1H), 7.39 (d, J = 8.0 Hz, 1H), 7.36−7.29 (m, 5H), 6.87 (d, J = 8.0 Hz, 1H), 6.76 (s, 1H), 5.90−5.79 (m, 1H), 5.79−5.69 (m, 1H), 5.26−5.04 (m, 5H), 5.03−4.98 (m, 1H), 4.84 (td, J = 7.0, 4.0 Hz, 1H), 4.79 (dt, J = 8.0, 3.0 Hz, 1H), 4.15−4.09 (m, 2H), 3.96−3.87 (m, 4H), 3.67 (dd, J = 9.5, 3.0 Hz, 1H), 3.52 (dd, J = 9.0, 6.5 Hz, 1H), 3.37 (s, 3H), 2.38 (s, 3H), 2.29−2.23 (m, 2H), 2.06−2.02 (m, 2H) ppm. ESI-MS: m/z = 539 [M + H]+. Benzyl O-Allyl-N-(N-(4-chloro-2-(pent-4-en-1-yloxy)benzoyl)-O-methyl-L-seryl)-L-serinate (21l). Obtained following the general procedure B from 20l and 10a. Viscous liquid (two steps, 0.39 g, 70%). 1H NMR (500 MHz, CDCl3) δ 8.70 (d, J = 7.0 Hz, 1H), 8.21 (dd, J = 8.5, 7.0 Hz, 1H), 7.37 (d, J = 9.0 Hz, 1H), 7.35− 7.31 (m, 5H), 6.76 (ddd, J = 9.0, 7.5, 2.5 Hz, 1H), 6.67 (dd, J = 10.5, 2.5 Hz, 1H), 5.88−5.70 (m, 2H), 5.26−5.05 (m, 5H), 5.04−5.00 (m, 1H), 4.84−4.76 (m, 2H), 4.12−4.08 (m, 2H), 3.96−3.87 (m, 4H), 3.67 (dd, J = 9.5, 3.5 Hz, 1H), 3.52 (dd, J = 9.0, 7.0 Hz, 1H), 3.38 (s, 3H), 2.29−2.23 (m, 2H), 2.10−2.01 (m, 2H) ppm. ESI-MS: m/z = 559 [M + H]+. Benzyl O-Allyl-N-(N-(5-chloro-2-(pent-4-en-1-yloxy)benzoyl)-O-methyl-L-seryl)-L-serinate (21m). Obtained following the general procedure B from 20m and 10a. White solid (two steps, 0.41 g, 73%); mp = 52−53 °C. 1H NMR (500 MHz, CDCl3) δ 8.80 (d, J = 6.5 Hz, 1H), 8.17 (d, J = 3.0 Hz, 1H), 7.40−7.30 (m, 7H), 6.90 (d, J = 9.0 Hz, 1H), 5.89−5.69 (m, 2H), 5.26−5.22 (m, 1H), 5.21−5.09 (m, 3H), 5.09−5.03 (m, 1H), 5.03−4.98 (m, 1H), 4.84− 4.76 (m, 2H), 4.13−4.09 (m, 2H), 3.95−3.87 (m, 4H), 3.67 (dd, J = 9.5, 3.5 Hz, 1H), 3.52 (dd, J = 9.0, 7.0 Hz, 1H), 3.37 (s, 3H), 2.28− 2.22 (m, 2H), 2.07−2.00 (m, 2H) ppm. 13C NMR (100 MHz, DMSO) δ 169.65, 169.40, 162.54, 155.67, 137.79, 135.78, 134.59, 132.44, 130.25, 128.30, 127.94, 127.64, 124.65, 122.62, 116.80, 115.32, 115.25, 72.30, 71.26, 69.02, 68.89, 66.09, 58.36, 52.62, 52.51, 29.57, 27.60. ESI-MS: m/z = 559 [M + H]+. Benzyl O-Allyl-N-(O-methyl-N-(3-(pent-4-en-1-yloxy)picolinoyl)-L-seryl)-L-serinate (21n). Obtained following the general procedure B from 20n and 10a. Viscous liquid (two steps, 0.50 g, 95%). 1H NMR (500 MHz, CDCl3) δ 8.58 (d, J = 7.0 Hz, 1H), 8.29 (dd, J = 4.0, 1.5 Hz, 1H), 7.42 (d, J = 8.0 Hz, 1H), 7.40− 7.28 (m, 7H), 5.88−5.71 (m, 2H), 5.26−5.10 (m, 4H), 5.10−4.97 (m, 2H), 4.83 (td, J = 7.0, 4.0 Hz, 1H), 4.78 (dt, J = 8.0, 3.5 Hz, 1H), 4.10 (t, J = 6.5 Hz, 2H), 3.95−3.89 (m, 4H), 3.68 (dd, J = 9.5, 3.5 Hz, 1H), 3.52 (dd, J = 9.0, 7.5 Hz, 1H), 3.37 (s, 3H), 2.31−2.24 (m, 2H), 2.06−1.99 (m, 2H) ppm. 13C NMR (100 MHz, CDCl3) δ 170.27, 169.80, 164.03, 155.18, 140.87, 137.45, 135.40, 134.00, 128.53, 128.39, 128.31, 128.09, 127.01, 121.52, 117.23, 115.52, 72.18, 71.82, 69.54, 68.67, 67.18, 59.05, 53.10, 52.60, 29.93, 27.99. ESI-MS: m/z = 526 [M + H]+. Benzyl O-Allyl-N-(N-(3-(hept-6-en-1-yloxy)benzoyl)-Omethyl-L-seryl)-L-serinate (21o). Obtained following the general procedure B from 20o and 10a. Viscous liquid (two steps, 0.50 g, 90%). 1H NMR (500 MHz, CDCl3) δ 7.51 (d, J = 8.0 Hz, 1H), 7.39− 7.30 (m, 8H), 7.12 (d, J = 6.0 Hz, 1H), 7.06−7.02 (m, 1H), 5.87− 5.72 (m, 2H), 5.29−5.12 (m, 4H), 5.05−4.99 (m, 1H), 4.97−4.93 (m, 1H), 4.80−4.72 (m, 2H), 4.00 (t, J = 6.5 Hz, 2H), 3.96−3.88 (m, 4H), 3.68 (dd, J = 9.5, 3.0 Hz, 1H), 3.49 (t, J = 8.5 Hz, 1H), 3.39 (s, 3H), 2.12−2.06 (m, 2H), 1.83−1.76 (m, 2H), 1.52−1.43 (m, 4H) ppm. 13C NMR (100 MHz, CDCl3) δ 170.44, 169.76, 167.14, 159.41, 138.80, 135.41, 135.19, 133.97, 129.58, 128.61, 128.44, 128.20, 118.98, 118.59, 117.37, 114.53, 113.10, 72.25, 71.73, 69.47, 68.16,

67.33, 59.10, 53.18, 52.48, 33.69, 29.10, 28.66, 25.55. ESI-MS: m/z = 553 [M + H]+. Benzyl O-Allyl-N-(O-methyl-N-(1-methyl-5-(pent-4-en-1yloxy)-1H-pyrazole-4-carbonyl)-L-seryl)-L-serinate (21p). Obtained following the general procedure B from 20p and 10a. Viscous liquid (two steps, 0.36 g, 68%). 1H NMR (500 MHz, CDCl3) δ 7.74 (s, 1H), 7.42 (d, J = 8.0 Hz, 1H), 7.36−7.32 (m, 5H), 7.02 (d, J = 6.5 Hz, 1H), 5.87−5.70 (m, 2H), 5.28−5.12 (m, 4H), 5.09−5.00 (m, 2H), 4.78−4.69 (m, 2H), 4.27−4.18 (m, 2H), 3.94−3.85 (m, 5H), 3.70 (s, 3H), 3.67 (dd, J = 9.5, 3.0 Hz, 1H), 3.47 (dd, J = 9.5, 7.5 Hz, 1H), 3.37 (s, 2H), 2.28−2.21 (m, 2H), 1.99−1.91 (m, 2H) ppm. 13C NMR (100 MHz, CDCl3) δ 170.45, 169.88, 162.04, 152.98, 140.55, 139.08, 137.37, 135.48, 134.06, 128.68, 128.51, 128.26, 117.43, 115.72, 75.87, 72.33, 71.97, 69.61, 67.40, 59.16, 53.22, 52.02, 34.56, 29.84, 28.93. ESI-MS: m/z = 529 [M + H]+. Benzyl O-Allyl-N-(O-methyl-N-(2-(pent-4-en-1-yloxy)nicotinoyl)-L-seryl)-L-serinate (21q). Obtained following the general procedure B from 20q and 10a. Viscous liquid (two steps, 0.31 g, 59%). 1H NMR (500 MHz, CDCl3) δ 8.87 (d, J = 6.5 Hz, 1H), 8.50 (dd, J = 7.5, 2.0 Hz, 1H), 8.27 (dd, J = 5.0, 2.0 Hz, 1H), 7.39−7.31 (m, 6H), 7.04 (dd, J = 7.5, 5.0 Hz, 1H), 5.91−5.81 (m, 1H), 5.79−5.70 (m, 1H), 5.27−5.04 (m, 5H), 5.01−4.97 (m, 1H), 4.84−4.78 (m, 2H), 4.58−4.46 (m, 2H), 3.96−3.87 (m, 4H), 3.68 (dd, J = 9.5, 3.5 Hz, 1H), 3.53 (dd, J = 9.0, 7.0 Hz, 1H), 3.39 (s, 3H), 2.29−2.22 (m, 2H), 2.05−1.98 (m, 2H) ppm. 13C NMR (100 MHz, CDCl3) δ 170.16, 169.89, 164.09, 160.93, 150.03, 141.67, 137.74, 135.49, 134.08, 128.62, 128.42, 128.19, 117.60, 117.25, 115.48, 115.44, 72.26, 71.81, 69.69, 67.30, 66.86, 59.16, 53.24, 53.16, 30.33, 28.12. ESI-MS: m/z = 526 [M + H]+. Benzyl O-Allyl-N-(O-methyl-N-(pent-4-enoyl-L -prolyl)- Lseryl)-L-serinate (21r). To a solution of 20r (0.20 mg, 1.0 mmol), HOBt (0.16 g,1.2 mmol) and EDCI (0.35 g,1.8 mmol) were dissolved in 3 mL of DCM and stirred at room temperature for 30 min. It was sequentially treated with 10a (0.45 g, 1.2 mmol) and DIPEA (0.50 mmol, 3.0 mmol). The reaction mixture was stirred at room temperature for 3 h and washed with saturated NaHCO3 (5 mL × 2) and brine (5 mL × 2), dried over Na2SO4, filtered, and concentrated in vacuo. The residue was then purified by chromatography on silica gel to give 21r (0.36 g, 70%) as a white solid; mp = 122−123 °C. 1H NMR (500 MHz, CDCl3) δ 7.40−7.29 (m, 6H), 7.26−7.24 (m, 1H), 5.92−5.72 (m, 2H), 5.29−5.11 (m, 4H), 5.09−4.95 (m, 2H), 4.78−4.70 (m, 1H), 4.60−4.48 (m, 2H), 3.98−3.91 (m, 2H), 3.89 (dd, J = 9.5, 3.5 Hz, 1H), 3.79 (dd, J = 9.5, 4.0 Hz, 1H), 3.67 (dd, J = 9.5, 3.5 Hz, 1H), 3.62−3.56 (m, 1H), 3.46 (dd, J = 9.0, 7.0 Hz, 2H), 3.33 (s, 3H), 2.50−2.35 (m, 4H), 2.32− 2.23 (m, 1H), 2.15−2.04 (m, 1H), 2.02−1.93 (m, 2H) ppm. 13C NMR (100 MHz, CDCl3) δ 172.31, 171.75, 169.96, 169.76, 137.31, 135.45, 134.09, 128.51, 128.27, 128.07, 117.15, 115.30, 72.15, 71.54, 69.52, 67.12, 59.99, 58.99, 53.00, 52.67, 47.49, 33.87, 28.75, 28.33, 24.94. ESI-MS: m/z = 516 [M + H]+. Benzyl O-Allyl-N-(O-methyl-N-(1-(pent-4-en-1-yl)-1H-pyrrole-2-carbonyl)-L-seryl)-L-serinate (21s). Obtained following the general procedure B from 20s and 10a. Viscous liquid (two steps, 0.25 g, 50%). 1H NMR (500 MHz, CDCl3) δ 7.48 (d, J = 8.0 Hz, 1H), 7.39−7.30 (m, 5H), 6.80−6.75 (m, 2H), 6.67 (dd, J = 4.0, 2.0 Hz, 1H), 6.10 (dd, J = 4.0, 2.5 Hz, 1H), 5.84−5.71 (m, 2H), 5.28−5.24 (m, 1H), 5.22−5.12 (m, 3H), 5.06−4.95 (m, 2H), 4.77 (dt, J = 8.0, 3.0 Hz, 1H), 4.70−4.65 (m, 1H), 4.40−4.26 (m, 2H), 3.97−3.90 (m, 3H), 3.87 (dd, J = 9.0, 4.0 Hz, 1H), 3.68 (dd, J = 9.5, 3.0 Hz, 1H), 3.46 (dd, J = 9.0, 8.0 Hz, 1H), 3.38 (s, 3H), 2.08−2.02 (m, 2H), 1.91−1.83 (m, 2H) ppm. ESI-MS: m/z = 498 [M + H]+. Benzyl O-Allyl-N-(O-methyl-N-(1-(pent-4-en-1-yl)-1H-imidazole-2-carbonyl)-L-seryl) -L-serinate (21t). Obtained following the general procedure B from 20t and 10a. Viscous liquid (two steps, 0.35 g, 70%). 1H NMR (500 MHz, CDCl3) δ 8.19 (d, J = 4.0 Hz, 1H), 7.38−7.29 (m, 6H), 7.07 (d, J = 1.0 Hz, 1H), 7.03 (d, J = 1.0 Hz, 1H), 5.85−5.70 (m, 2H), 5.26−5.22 (m, 1H), 5.21−5.10 (m, 3H), 5.08−4.99 (m, 2H), 4.79 (dt, J = 8.0, 3.0 Hz, 1H), 4.69 (td, J = 7.0, 4.5 Hz, 1H), 4.48−4.42 (m, 2H), 3.98−3.85 (m, 4H), 3.67 (dd, J = 9.5, 3.0 Hz, 1H), 3.57 (dd, J = 9.0, 6.5 Hz, 1H), 3.38 (s, 3H), 2.81 (s, 9193



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3H), 2.12−2.06 (m, 2H), 1.97−1.88 (m, 2H) ppm. 13C NMR (100 MHz, CDCl3) δ 170.20, 169.79, 165.82, 160.07, 141.26, 137.11, 133.96, 132.33, 128.65, 128.50, 128.23, 125.05, 117.45, 115.83, 72.28, 71.80, 69.49, 67.40, 59.16, 53.20, 52.12, 46.45, 30.54, 30.14. ESI-MS: m/z = 499 [M + H]+. Benzyl O-Allyl-N-(N-(1-(hex-5-en-1-yl)-1H-imidazole-2-carbonyl)-O-methyl-L-seryl)-L-serinate (21u). Obtained following the general procedure B from 20u and 10a. Viscous liquid (two steps, 0.30 g, 59%). 1H NMR (500 MHz, CDCl3) δ 8.09 (d, J = 7.5 Hz, 1H), 7.37−7.29 (m, 6H), 7.04 (d, J = 1.0 Hz, 1H), 7.01 (d, J = 1.0 Hz, 1H), 5.81−5.67 (m, 2H), 5.26−5.09 (m, 4H), 5.02−4.92 (m, 2H), 4.78 (dt, J = 8.0, 3.0 Hz, 1H), 4.69 (td, J = 6.5, 4.0 Hz, 1H), 4.44 (t, J = 7.5 Hz, 2H), 3.96−3.84 (m, 4H), 3.66 (dd, J = 9.5, 3.5 Hz, 1H), 3.55 (dd, J = 9.5, 7.0 Hz, 1H), 3.37 (s, 3H), 2.10−2.04 (m, 2H), 1.85−1.77 (m, 2H), 1.45−1.37 (m, 2H) ppm. 13C NMR (100 MHz, CDCl3) δ 169.79, 159.12, 138.14, 138.00, 135.41, 134.00, 128.53, 128.31, 128.06, 124.50, 117.23, 114.94, 72.18, 71.73, 69.54, 67.17, 59.09, 53.06, 52.37, 48.07, 33.17, 30.72, 25.78. ESI-MS: m/z = 513 [M + H]+. Benzyl O-Allyl-N-(O-methyl-N-(1-(pent-4-en-1-yl)-1H-pyrazole-5-carbonyl)-L-seryl) -L-serinate (21v). Obtained following the general procedure B from 20v and 10a. White solid (two steps, 0.33 g, 66%); mp = 73−74 °C. 1H NMR (500 MHz, CDCl3) δ 7.50− 7.45 (m, 2H), 7.39−7.31 (m, 5H), 6.96 (d, J = 6.5 Hz, 1H), 6.59 (d, J = 2.0 Hz, 1H), 5.85−5.72 (m, 2H), 5.29−5.12 (m, 4H), 5.06−4.94 (m, 2H), 4.76 (dt, J = 8.5, 3.0 Hz, 1H), 4.67 (ddd, J = 8.0, 6.0, 4.0 Hz, 1H), 4.60−4.52 (m, 2H), 3.99−3.90 (m, 3H), 3.86 (dd, J = 9.0, 4.0 Hz, 1H), 3.68 (dd, J = 9.5, 3.5 Hz, 1H), 3.47 (t, J = 8.5 Hz, 1H), 3.39 (s, 3H), 2.10−2.05 (m, 2H), 1.99−1.92 (m, 2H) ppm. 13C NMR (100 MHz, CDCl3) δ 170.02, 169.69, 159.68, 137.77, 137.61, 135.34, 134.33, 133.90, 128.61, 128.46, 128.18, 117.39, 115.12, 107.09, 72.22, 71.61, 69.41, 67.36, 59.11, 53.17, 52.05, 51.10, 30.73, 29.78. ESI-MS: m/z = 499 [M + H]+. Benzyl O-Allyl-N-(N-(1-(hex-5-en-1-yl)-1H-pyrazole-5-carbonyl)-O-methyl-L-seryl)-L-serinate (21w). Obtained following the general procedure B from 20w and 10a. White solid (two steps, 0.48 g, 93%); mp = 62−63 °C. 1H NMR (500 MHz, CDCl3) δ 7.49 (d, J = 8.5 Hz, 1H), 7.46 (d, J = 2.0 Hz, 1H), 7.38−7.30 (m, 5H), 7.00 (d, J = 6.5 Hz, 1H), 6.59 (d, J = 2.0 Hz, 1H), 5.81−5.72 (m, 2H), 5.28−5.12 (m, 4H), 5.01−4.90 (m, 2H), 4.75 (dt, J = 8.0, 3.0 Hz, 1H), 4.67 (ddd, J = 8.0, 6.5, 4.0 Hz, 1H), 4.54 (t, J = 7.5 Hz, 2H), 3.98−3.88 (m, 3H), 3.85 (dd, J = 9.0, 4.0 Hz, 1H), 3.67 (dd, J = 9.5, 3.0 Hz, 1H), 3.47 (t, J = 8.5 Hz, 1H), 3.38 (s, 3H), 2.09−2.04 (m, 2H), 1.88−1.81 (m, 2H), 1.43−1.35 (m, 2H) ppm. 13C NMR (100 MHz, CDCl3) δ 169.94, 169.53, 159.55, 138.26, 137.47, 135.21, 134.11, 133.77, 128.40, 128.24, 127.95, 117.13, 114.51, 106.96, 71.99, 71.56, 69.26, 67.09, 58.86, 52.98, 51.93, 51.21, 33.09, 29.97, 25.66. ESI-MS: m/z = 513 [M + H]+. Benzyl N-(N-(5-Allyl-1-methyl-1H-pyrazole-4-carbonyl)-Omethyl-L-seryl)-O-(hex-5-en-1-yl)-L-serinate (21x). Obtained following the general procedure B from 20x and 10c. Viscous liquid (two steps, 0.36 g, 68%). 1H NMR (500 MHz, CDCl3) δ 7.72 (s, 1H), 7.47 (d, J = 8.0 Hz, 1H), 7.38−7.29 (m, 5H), 6.72 (d, J = 6.0 Hz, 1H), 5.89 (ddt, J = 17.0, 10.0, 6.0 Hz, 1H), 5.81−5.72 (m, 1H), 5.26−5.21 (m, 1H), 5.18−5.09 (m, 2H), 5.02−4.92 (m, 3H), 4.74 (dt, J = 8.0, 3.0 Hz, 1H), 4.69 (ddd, J = 8.0, 6.5, 4.0 Hz, 1H), 3.89 (dd, J = 9.5, 3.0 Hz, 1H), 3.85 (dd, J = 9.0, 4.0 Hz, 1H), 3.81−3.77 (m, 5H), 3.65 (dd, J = 9.5, 3.5 Hz, 1H), 3.48−3.43 (m, 1H), 3.42− 3.34 (m, 5H), 2.06−1.97 (m, 2H), 1.54−1.45 (m, 2H), 1.42−1.31 (m, 2H) ppm. ESI-MS: m/z = 527 [M + H]+. Methyl O-Allyl-N-(O-methyl-N-(3-(pent-4-en-1-yloxy)thiophene-2-carbonyl)-L-seryl)-L-serinate (21y). Obtained following the general procedure B from 20y and 10b. Viscous liquid (two steps, 0.27 g, 84%). 1H NMR (500 MHz, CDCl3) δ 8.13 (d, J = 7.0 Hz, 1H), 7.39 (d, J = 5.5 Hz, 1H), 7.36 (d, J = 8.5 Hz, 1H), 6.83 (d, J = 5.5 Hz, 1H), 5.88−5.76 (m, 2H), 5.24−5.18 (m, 1H), 5.16− 5.12 (m, 1H), 5.10−5.04 (m, 1H), 5.03−4.98 (m, 1H), 4.79−4.71 (m, 2H), 4.18 (td, J = 6.5, 2.0 Hz, 2H), 3.98−3.94 (m, 3H), 3.89 (dd, J = 9.5, 3.0 Hz, 1H), 3.75 (s, 3H), 3.66 (dd, J = 9.5, 3.5 Hz, 1H), 3.55

(dd, J = 9.0, 7.0 Hz, 1H), 3.43 (s, 3H), 2.30−2.24 (m, 2H), 2.01− 1.93 (m, 2H) ppm. ESI-MS: m/z = 455 [M + H]+. Methyl O-Allyl-N-(O-methyl-N-(2-methyl-4-(pent-4-en-1yloxy)thiazole-5-carbonyl)-L-seryl)-L-serinate (21z). Obtained following the general procedure B from 20z and 10b. Viscous liquid (two steps, 0.27 g, 84%). 1H NMR (500 MHz, CDCl3) δ 7.93 (d, J = 7.0 Hz, 1H), 7.34 (d, J = 8.0 Hz, 1H), 5.89−5.76 (m, 2H), 5.25−5.17 (m, 1H), 5.17−5.12 (m, 1H), 5.10−5.04 (m, 1H), 5.02−4.97 (m, 1H), 4.76−4.68 (m, 2H), 4.53−4.44 (m, 2H), 3.99−3.95 (m, 2H), 3.93 (dd, J = 9.0, 4.0 Hz, 1H), 3.89 (dd, J = 9.5, 3.0 Hz, 1H), 3.76 (s, 3H), 3.66 (dd, J = 9.5, 3.0 Hz, 1H), 3.54 (dd, J = 9.0, 7.0 Hz, 1H), 3.43 (s, 3H), 2.61 (s, 3H), 2.28−2.21 (m, 2H), 1.97−1.90 (m, 2H) ppm. 13C NMR (100 MHz, CDCl3) δ 170.58, 170.29, 167.21, 161.33, 160.78, 137.63, 134.19, 117.36, 115.56, 106.80, 72.36, 71.97, 70.90, 69.65, 59.25, 53.08, 52.94, 52.66, 30.01, 28.70, 20.33. ESI-MS: m/z = 470 [M + H]+. Benzyl (S)-2-((S)-3-Methoxy-2-(2-(pent-4-en-1-yloxy)benzamido)propanamido)hex-5-enoate (21za). Obtained following the general procedure B from 20b and 10d. Viscous liquid (two steps, 0.45 g, 88%). 1H NMR (500 MHz, CDCl3) δ 8.83 (d, J = 7.0 Hz, 1H), 8.21 (dd, J = 8.0, 1.5 Hz, 1H), 7.50−7.42 (m, 1H), 7.39−7.29 (m, 5H), 7.14 (d, J = 8.0 Hz, 1H), 7.07 (t, J = 7.5 Hz, 1H), 6.97 (d, J = 8.5 Hz, 1H), 5.88−5.67 (m, 2H), 5.21−5.10 (m, 2H), 5.09−4.90 (m, 4H), 4.81 (td, J = 6.5, 4.0 Hz, 1H), 4.69 (td, J = 7.5, 4.5 Hz, 1H), 4.16−4.12 (m, 2H), 3.96 (dd, J = 9.0, 3.5 Hz, 1H), 3.51 (dd, J = 9.0, 6.0 Hz, 1H), 3.39 (s, 3H), 2.29−2.22 (m, 2H), 2.11− 1.93 (m, 5H), 1.84−1.73 (m, 1H) ppm. ESI-MS: m/z = 509 [M + H]+. Benzyl (S)-2-((S)-2-(2-(Hex-5-en-1-yloxy)benzamido)-3methoxypropanamido)hex-5-enoate (21zb). Obtained following the general procedure B from 20c and 10d. Viscous liquid (two steps, 0.45 g, 86%). 1H NMR (500 MHz, CDCl3) δ 8.83 (d, J = 6.5 Hz, 1H), 8.21 (dd, J = 8.0, 1.5 Hz, 1H), 7.45 (ddd, J = 8.5, 7.0, 2.0 Hz, 1H), 7.38−7.29 (m, 5H), 7.15 (d, J = 8.0 Hz, 1H), 7.10−7.04 (m, 1H), 6.97 (d, J = 8.0 Hz, 1H), 5.86−5.67 (m, 2H), 5.21−5.10 (m, 2H), 5.06−4.90 (m, 4H), 4.81 (td, J = 6.5, 3.5 Hz, 1H), 4.68 (td, J = 7.5, 4.5 Hz, 1H), 4.16−4.11 (m, 2H), 3.94 (dd, J = 9.0, 3.5 Hz, 1H), 3.51 (dd, J = 9.0, 6.5 Hz, 1H), 3.40 (s, 3H), 2.17−2.10 (m, 2H), 2.09−1.90 (m, 5H), 1.84−1.75 (m, 1H), 1.63−1.54 (m, 2H) ppm. 13 C NMR (100 MHz, CDCl3) δ 171.81, 170.23, 165.62, 157.48, 138.31, 136.97, 135.42, 133.25, 132.39, 128.65, 128.48, 128.32, 121.13, 120.84, 115.74, 115.04, 112.29, 71.81, 69.17, 67.14, 59.12, 53.29, 52.13, 33.37, 31.66, 29.40, 28.54, 25.41. ESI-MS: m/z = 523[M + H]+. (8S,11S)-3,4,5,7,8,9,10,11,12,13-decahydro-2H-11-(Methoxymethyl)-10,13-dioxo-1,6,9,12-benzodioxadiazacyclopentadecine-8-carboxylic Acid (22a). Obtained following the general procedure C from 21a. White solid (58 mg, 76%); mp 168−169 °C. 1 H NMR (500 MHz, CDCl3) δ 8.48 (d, J = 9.0 Hz, 1H), 8.09 (d, J = 7.0 Hz, 1H), 7.50−7.43 (m, 1H), 7.35 (d, J = 7.0 Hz, 1H), 7.12−7.06 (m, 1H), 6.99 (d, J = 8.5 Hz, 1H), 5.04−4.97 (m, 1H), 4.74−4.68 (m, 1H), 4.23−4.03 (m, 4H), 3.74−3.66 (m, 2H), 3.51 (dd, J = 9.0, 3.5 Hz, 1H), 3.45−3.40 (m, 1H), 3.35 (s, 3H), 2.06−1.97 (m, 1H), 1.87−1.75 (m, 2H), 1.69−1.60 (m, 1H) ppm. 13C NMR (100 MHz, CDCl3) δ 171.57, 170.17, 166.46, 157.40, 133.49, 132.17, 121.64, 113.35, 72.05, 71.65, 71.57, 69.66, 59.36, 53.67, 53.50, 27.13, 25.70. ESI-MS: m/z = 381 [M + H]+. (9S,12S)-2,3,4,5,6,8,9,10,11,12,13,14-Dodecahydro-12-(methoxymethyl)-11,14-dioxo-1,7,10,13-benzodioxadiazacyclohexadecine-9-carboxylic Acid (22b). Obtained following the general procedure C from 21b. White solid (66 mg, 84%); mp = 166−167 °C. 1H NMR (500 MHz, CDCl3) δ 8.93 (d, J = 9.0 Hz, 1H), 8.19 (d, J = 7.0 Hz, 1H), 7.50−7.41 (m, 1H), 7.28 (d, J = 9.0 Hz, 1H), 7.11−7.03 (m, 1H), 6.98 (d, J = 8.5 Hz, 1H), 5.00−4.91 (m, 1H), 4.80−4.71 (m, 1H), 4.27−4.20 (m, 1H), 4.17−4.08 (m, 2H), 4.06−3.96 (m, 1H), 3.79−3.73 (m, 1H), 3.67−3.61 (m, 1H), 3.59− 3.53 (m, 1H), 3.50 (dd, J = 9.0, 3.5 Hz, 1H), 3.37 (s, 3H), 1.93−1.73 (m, 3H), 1.60−1.48 (m, 3H) ppm. 13C NMR (100 MHz, CDCl3) δ 171.89, 170.04, 166.38, 157.63, 133.68, 132.60, 121.25, 120.38, 9194



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112.19, 72.32, 70.97, 70.19, 69.17, 59.21, 53.80, 53.02, 28.90, 28.51, 24.30. ESI-MS: m/z = 395 [M + H]+. (10S,13S)-3,4,5,6,7,9,10,11,12,13,14,15-Dodecahydro-2H13-(methoxymethyl)-12,15-dioxo-1,8,11,14-benzodioxadiazacycloheptadecine-10-carboxylic Acid (22c). Obtained following the general procedure C from 21c. White solid (62 mg, 76%); mp = 102−103 °C. 1H NMR (500 MHz, CDCl3) δ 9.00 (d, J = 8.0 Hz, 1H), 8.18 (d, J = 7.0 Hz, 1H), 7.47−7.38 (m, 1H), 7.23 (d, J = 6.5 Hz, 1H), 7.06−6.99 (m, 1H), 6.94 (d, J = 8.5 Hz, 1H), 4.92−4.82 (m, 1H), 4.65−4.55 (m, 1H), 4.23−4.16 (m, 1H), 4.14−4.05 (m, 2H), 3.80−3.67 (m, 2H), 3.56−3.47 (m, 2H), 3.45−3.40 (m, 1H), 3.36 (s, 3H), 1.98−1.88 (m, 1H), 1.82−1.69 (m, 2H), 1.63−1.36 (m, 5H) ppm. 13C NMR (100 MHz, CDCl3) δ 171.74, 170.39, 166.25, 157.75, 133.73, 132.61, 121.09, 119.82, 111.88, 72.07, 71.49, 69.41, 69.30, 59.23, 53.94, 52.96, 29.74, 28.70, 27.94, 27.22. ESI-MS: m/z = 409 [M + H]+. (11S,14S)-2,3,4,5,6,7,8,10,11,12,13,14,15,16-Tetradecahydro-14-(methoxymethyl)-13,16-dioxo-1,9,12,15-benzodioxadiazacyclooctadecine-11-carboxylic Acid (22d). Obtained following the general procedure C from 21d. White solid (68 mg, 80%). 1 H NMR (500 MHz, CDCl3) δ 9.00 (d, J = 7.5 Hz, 1H), 8.18 (d, J = 7.5 Hz, 1H), 7.50−7.40 (m, 1H), 7.23 (d, J = 6.5 Hz, 1H), 7.10−7.01 (m, 1H), 6.98 (d, J = 8.5 Hz, 1H), 4.87−4.81 (m, 1H), 4.75−4.69 (m, 1H), 4.28−4.21 (m, 1H), 4.15−4.03 (m, 2H), 3.86 (dd, J = 10.5, 4.5 Hz, 1H), 3.76 (dd, J = 10.5, 2.5 Hz, 1H), 3.56 (dd, J = 9.0, 4.5 Hz, 1H), 3.53−3.47 (m, 1H), 3.45−3.35 (m, 4H), 2.03−1.94 (m, 1H), 1.88−1.77 (m, 1H), 1.64−1.32 (m, 8H) ppm. ESI-MS: m/z = 423 [M + H]+. (12S,15S)-3,4,5,6,7,8,9,11,12,13,14,15,16,17-Tetradecahydro-2H-15-methoxymethyl)-14,17-dioxo-1,10,13,16-benzodioxadiazacyclononadecine-12-carboxylic Acid (22e). Obtained following the general procedure C from 21e. White solid (67 mg, 77%). 1H NMR (500 MHz, CDCl3) δ 9.03 (d, J = 6.0 Hz, 1H), 8.20 (d, J = 7.5 Hz, 1H), 7.49−7.39 (m, 1H), 7.28 (d, J = 7.5 Hz, 1H), 7.07−7.00 (m, 1H), 6.97 (d, J = 8.5 Hz, 1H), 4.80−4.75 (m, 1H), 4.74−4.68 (m, 1H), 4.22−4.12 (m, 2H), 3.97 (dd, J = 9.5, 4.0 Hz, 1H), 3.80−3.70 (m, 2H), 3.66 (dd, J = 9.5, 5.0 Hz, 1H), 3.54− 3.48 (m, 1H), 3.46−3.36 (m, 4H), 2.03−1.96 (m, 1H), 1.91−1.82 (m, 1H), 1.60−1.28 (m, 10H) ppm. ESI-MS: m/z = 437 [M + H]+. (13S,16S)-3,4,5,6,7,8,9,10,13,14,15,16,17,18-Tetradecahydro-2H,12H-16 (methoxymethyl)-15,18-dioxo-benzo[i][1,11]dioxa[4,7]diazacycloicosine-13-carboxylic Acid (22f). Obtained following the general procedure C from 21f. White solid (67 mg, 77%). 1H NMR (500 MHz, CDCl3) δ 8.97 (d, J = 6.5 Hz, 1H), 8.21 (d, J = 7.5 Hz, 1H), 7.49−7.41 (m, 1H), 7.22 (d, J = 6.0 Hz, 1H), 7.07−7.01 (m, 1H), 6.97 (d, J = 8.5 Hz, 1H), 4.82−4.75 (m, 1H), 4.68−4.61 (m, 1H), 4.23−4.18 (m, 1H), 4.17−4.12 (m, 1H), 4.01 (dd, J = 9.5, 3.0 Hz, 1H), 3.80−3.72 (m, 2H), 3.63 (dd, J = 9.5, 3.5 Hz, 1H), 3.55−3.48 (m, 1H), 3.41−3.29 (m, 4H), 2.01−1.89 (m, 1H), 1.83−1.72 (m, 1H), 1.67−1.29 (m, 12H) ppm. ESI-MS: m/z = 451 [M + H]+. (10S,13S)-19-Fluoro-13-(methoxymethyl)-12,15-dioxo2,3,4,5,6,7,10,11,12,13,14,15-dodecahydro-9H-benzo[i][1,11]dioxa[4,7]diazacycloheptadecine-10-carboxylic Acid (22g). Obtained following the general procedure C from 21g. White solid (70 mg, 82%). 1H NMR (500 MHz, CDCl3) δ 8.96 (d, J = 8.0 Hz, 1H), 7.92 (d, J = 8.0 Hz, 1H), 7.25−7.20 (m, 2H), 7.13−7.02 (m, 1H), 4.88−4.79 (m, 1H), 4.68−4.60 (m, 1H), 4.42−4.34 (m, 1H), 4.32−4.24 (m, 1H), 4.13−4.06 (m, 1H), 3.80 (dd, J = 9.5, 3.5 Hz, 1H), 3.75 (dd, J = 9.5, 2.5 Hz, 1H), 3.57−3.49 (m, 2H), 3.39−3.34 (m, 4H), 2.01−1.91 (m, 1H), 1.77−1.68 (m, 1H), 1.64−1.36 (m, 6H) ppm. ESI-MS: m/z = 427 [M + H]+. (10S,13S)-19-Methoxy-13-(methoxymethyl)-12,15-dioxo 2,3,4,5,6,7,10,11,12,13,14, 15-dodecahydro-9H-benzo[i][1,11]dioxa[4,7]diazacycloheptadecine-10-carboxylic Acid (22h). Obtained following the general procedure C from 21h. White solid (60 mg, 68%). 1H NMR (500 MHz, CDCl3) δ 9.14 (d, J = 7.5 Hz, 1H), 7.72 (d, J = 7.5 Hz, 1H), 7.23 (d, J = 7.0 Hz, 1H), 7.15−7.10 (m, 1H), 7.10−7.04 (m, 1H), 4.85−4.78 (m, 1H), 4.69− 4.61 (m, 1H), 4.11−4.01 (m, 2H), 3.91−3.81 (m, 5H), 3.75 (dd, J = 9.5, 2.5 Hz, 1H), 3.54 (dd, J = 9.0, 3.5 Hz, 2H), 3.42−3.32 (m, 4H),

1.78−1.67 (m, 1H), 1.64−1.55 (m, 1H), 1.53−1.18 (m, 6H) ppm. ESI-MS: m/z = 439 [M + H]+. (10S,13S)-18-Fluoro-13-(methoxymethyl)-12,15-dioxo2,3,4,5,6,7,10,11,12,13,14,15-dodecahydro-9H-benzo[i][1,11]dioxa[4,7]diazacycloheptadecine-10-carboxylic Acid (22i). Obtained following the general procedure C from 21i. White solid (59 mg, 69%). 1H NMR (500 MHz, CDCl3) δ 8.87 (d, J = 8.5 Hz, 1H), 8.15 (d, J = 8.5 Hz, 1H), 7.15 (d, J = 7.0 Hz, 1H), 7.05 (d, J = 8.5 Hz, 1H), 6.98 (s, 1H), 4.93−4.86 (m, 1H), 4.72−4.64 (m, 1H), 4.24−4.18 (m, 1H), 4.13−4.08 (m, 2H), 3.82−3.70 (m, 2H), 3.56− 3.49 (m, 2H), 3.41−3.32 (m, 5H), 2.00−1.89 (m, 1H), 1.83−1.72 (m, 2H), 1.67−1.35 (m, 5H) ppm. 13C NMR (100 MHz, DMSO) δ 170.98, 169.09, 164.97 (d, 1JC−F = 248 Hz), 163.42, 158.83 (d, 3JC−F = 11 Hz), 133.47 (d, 3JC−F = 10 Hz), 117.00, 107.41 (d, 2JC−F = 22 Hz), 100.82 (d, 2JC−F = 26 Hz), 71.88, 69.95, 69.91, 68.84, 58.64, 52.93, 52.35, 28.90, 28.38, 26.70, 26.30. ESI-MS: m/z = 427 [M + H]+. (10S,13S)-18-Methoxy-13-(methoxymethyl)-12,15-dioxo2,3,4,5,6,7,10,11,12,13,14, 15-dodecahydro-9H-benzo[i][1,11]dioxa[4,7]diazacycloheptadecine-10-carboxylic Acid (22j). Obtained following the general procedure C from 21j. White solid (59 mg, 67%). 1H NMR (500 MHz, CDCl3) δ 8.87 (d, J = 8.0 Hz, 1H), 8.16 (d, J = 8.5 Hz, 1H), 7.18 (d, J = 6.5 Hz, 1H), 6.58 (d, J = 8.5 Hz, 1H), 6.47 (s, 1H), 4.93−4.84 (m, 1H), 4.69−4.59 (m, 1H), 4.22−4.16 (m, 1H), 4.14−4.05 (m, 2H), 3.85 (s, 3H), 3.79 (dd, J = 9.5, 4.5 Hz, 1H), 3.73 (dd, J = 9.5, 3.0 Hz, 1H), 3.57−3.48 (m, 2H), 3.43−3.31 (m, 4H), 2.00−1.88 (m, 1H), 1.84−1.71 (m, 2H), 1.69− 1.36 (m, 5H) ppm. 13C NMR (100 MHz, CDCl3) δ 171.63, 170.99, 166.25, 164.22, 159.15, 134.45, 113.11, 105.47, 99.14, 72.19, 71.36, 69.40, 69.22, 59.32, 55.72, 54.02, 52.95, 29.69, 28.88, 27.79, 27.33. ESI-MS: m/z = 439 [M + H]+. (10S,13S)-13-(Methoxymethyl)-18-methyl-12,15-dioxo2,3,4,5,6,7,10,11,12,13,14,15-dodecahydro-9H-benzo[i][1,11]dioxa[4,7]diazacycloheptadecine-10-carboxylic Acid (22k). Obtained following the general procedure C from 21k. White solid (63 mg, 75%); mp = 169−170 °C. 1H NMR (500 MHz, CDCl3) δ 8.98 (d, J = 8.5 Hz, 1H), 8.09 (d, J = 8.0 Hz, 1H), 7.16 (d, J = 7.0 Hz, 1H), 6.87 (d, J = 8.0 Hz, 1H), 6.77 (s, 1H), 4.96−4.89 (m, 1H), 4.72−4.64 (m, 1H), 4.23−4.15 (m, 1H), 4.15−4.08 (m, 2H), 3.78 (dd, J = 10.0, 4.5 Hz, 1H), 3.73 (dd, J = 10.0, 3.5 Hz, 1H), 3.56−3.47 (m, 2H), 3.43−3.33 (m, 4H), 2.38 (s, 3H), 1.99−1.89 (m, 1H), 1.82−1.72 (m, 2H), 1.66−1.36 (m, 5H) ppm. 13C NMR (100 MHz, CDCl3) δ 171.68, 170.51, 166.42, 157.72, 144.70, 132.59, 121.97, 117.19, 112.58, 72.11, 71.42, 69.27, 59.23, 53.88, 52.93, 29.77, 28.75, 27.88, 27.24, 21.89. ESI-MS: m/z = 423 [M + H]+. (10S,13S)-18-Chloro-13-(methoxymethyl)-12,15-dioxo2,3,4,5,6,7,10,11,12,13,14,15-dodecahydro-9H-benzo[i][1,11]dioxa[4,7]diazacycloheptadecine-10-carboxylic Acid (22l). Obtained following the general procedure C from 21l. White solid (63 mg, 75%). 1H NMR (500 MHz, CDCl3) δ 8.86 (d, J = 8.0 Hz, 1H), 8.26−8.18 (m, 1H), 7.15 (d, J = 6.5 Hz, 1H), 6.79−6.72 (m, 1H), 6.69 (d, J = 10.5 Hz, 1H), 4.93−4.86 (m, 1H), 4.72−4.64 (m, 1H), 4.23−4.16 (m, 1H), 4.14−4.04 (m, 2H), 3.82−3.69 (m, 2H), 3.57−3.49 (m, 2H), 3.41−3.32 (m, 4H), 1.99−1.89 (m, 1H), 1.82− 1.73 (m, 2H), 1.66−1.36 (m, 5H) ppm. 13C NMR (100 MHz, DMSO) δ 170.99, 169.06, 163.44, 157.84, 137.60, 132.79, 120.67, 119.37, 113.11, 71.85, 69.90, 68.83, 58.64, 53.00, 52.33, 28.94, 28.40, 26.67, 26.28. ESI-MS: m/z = 443 [M + H]+. (10S,13S)-17-Chloro-13-(methoxymethyl)-12,15-dioxo2,3,4,5,6,7,10,11,12,13,14,15-dodecahydro-9H-benzo[i][1,11]dioxa[4,7]diazacycloheptadecine-10-carboxylic Acid (22m). Obtained following the general procedure C from 21m. White solid (61 mg, 69%). 1H NMR (500 MHz, CDCl3) δ 8.95 (d, J = 8.5 Hz, 1H), 8.18 (d, J = 3.0 Hz, 1H), 7.39 (dd, J = 8.5, 3.0 Hz, 1H), 7.13 (d, J = 7.0 Hz, 1H), 6.91 (d, J = 8.5 Hz, 1H), 4.93−4.86 (m, 1H), 4.75− 4.66 (m, 1H), 4.27−4.18 (m, 1H), 4.14−4.07 (m, 2H), 3.81 (dd, J = 10.0, 4.5 Hz, 1H), 3.75 (dd, J = 10.0, 3.5 Hz, 1H), 3.58−3.52 (m, 2H), 3.42−3.34 (m, 4H), 2.00−1.90 (m, 1H), 1.85−1.72 (m, 2H), 1.68−1.38 (m, 5H) ppm. ESI-MS: m/z = 443 [M + H]+. (14S,17S)-17-(Methoxymethyl)-16,19-dioxo6,7,8,9,10,11,14,15,16,17,18,19-dodecahydro-13H-pyrido[2,39195



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i][1,11]dioxa[4,7]diazacycloheptadecine-14-carboxylic Acid (22n). Obtained following the general procedure C from 21n. Brown solid (45 mg, 55%). 1H NMR (500 MHz, CDCl3) δ 8.81 (s, 1H), 8.44−8.26 (m, 1H), 7.59−7.30 (m, 3H), 4.95−4.85 (m, 1H), 4.65−4.52 (m, 1H), 4.27−4.08 (m, 3H), 3.86−3.77 (m, 1H), 3.76− 3.67 (m, 1H), 3.63−3.55 (m, 1H), 3.51 (s, 1H), 3.44−3.30 (m, 4H), 1.99−1.86 (m, 1H), 1.86−1.72 (m, 2H), 1.64−1.36 (m, 5H) ppm. ESI-MS: m/z = 410 [M + H]+. (13S,16S)-16-(Methoxymethyl)-15,18-dioxo-2,11-dioxa14,17-diazabicyclo[17.3.1]- tricosa-1(23),19,21-triene-13-carboxylic Acid (22o). Obtained following the general procedure C from 21o. White solid (61 g, 70%). 1H NMR (500 MHz, CDCl3) δ 7.42 (d, J = 8.0 Hz, 1H), 7.39−7.31 (m, 3H), 7.09−7.00 (m, 2H), 4.92−4.84 (m, 1H), 4.74−4.68 (m, 1H), 4.19−4.09 (m, 2H), 4.07 (dd, J = 10.0, 3.5 Hz, 1H), 4.03−3.93 (m, 1H), 3.88 (dd, J = 10.0, 3.5 Hz, 1H), 3.67 (dd, J = 10.0, 3.5 Hz, 1H), 3.62 (dd, J = 9.5, 4.0 Hz, 1H), 3.44−3.36 (m, 4H), 1.86−1.69 (m, 3H), 1.55−1.40 (m, 5H), 1.40−1.29 (m, 4H) ppm. ESI-MS: m/z = 437 [M + H]+. (6S,9S)-6-(Methoxymethyl)-1-methyl-4,7-dioxo1,4,5,6,7,8,9,10,12,13,14,15,16,17 tetradecahydropyrazolo[4,3-i][1,11]dioxa[4,7]diazacycloheptadecine-9-carboxylic Acid (22p). Obtained following the general procedure C from 21p. White solid (50 mg, 61%). 13C NMR (100 MHz, CDCl3) δ 171.68, 170.16, 163.01, 153.04, 139.68, 102.81, 71.82, 71.69, 69.78, 59.30, 53.44, 53.16, 34.77, 28.86, 27.89, 26.27, 24.32. ESI-MS: m/z = 413 [M + H]+. (7S,10S)-7-(Methoxymethyl)-5,8-dioxo6,7,8,9,10,11,13,14,15,16,17,18-dodecahydro-5H-pyrido[3,2i][1,11]dioxa[4,7]diazacycloheptadecine-10-carboxylic Acid (22q). Obtained following the general procedure C from 21q. Brown solid (68 mg, 82%). 1H NMR (500 MHz, CDCl3) δ 9.00 (d, J = 8.5 Hz, 1H), 8.51 (dd, J = 7.5, 2.0 Hz, 1H), 8.29 (dd, J = 5.0, 2.0 Hz, 1H), 7.14 (d, J = 7.0 Hz, 1H), 7.06 (dd, J = 7.5, 5.0 Hz, 1H), 4.93−4.87 (m, 1H), 4.71−4.65 (m, 1H), 4.62 (td, J = 10.5, 2.0 Hz, 1H), 4.42 (ddd, J = 10.5, 4.5, 3.0 Hz, 1H), 4.16−4.09 (m, 1H), 3.81 (dd, J = 10.0, 4.0 Hz, 1H), 3.73 (dd, J = 10.0, 3.5 Hz, 1H), 3.58−3.50 (m, 2H), 3.43−3.37 (m, 4H), 1.95−1.85 (m, 1H), 1.83−1.67 (m, 2H), 1.65−1.47 (m, 4H), 1.46−1.36 (m, 1H) ppm. 13C NMR (100 MHz, DMSO) δ 171.04, 168.96, 163.03, 160.44, 150.06, 140.89, 117.64, 115.15, 71.84, 69.84, 68.90, 67.45, 58.65, 52.95, 52.60, 28.86, 28.55, 26.62, 26.29. ESI-MS: m/z = 410 [M + H]+. (3S,6S,18aS)-3-(Methoxymethyl)-1,4,14-trioxohexadecahydro-1H-pyrrolo[2,1-i][1] oxa[4,7,10]triazacyclohexadecine-6carboxylic Acid (22r). Obtained following the general procedure C from 21r. Light-yellow oil (68 mg, 85%). 1H NMR (500 MHz, CDCl3) δ 7.68 (d, J = 7.0 Hz, 1H), 6.65 (d, J = 8.0 Hz, 1H), 4.79− 4.69 (m, 1H), 4.59−4.44 (m, 2H), 3.98 (dd, J = 10.0, 2.5 Hz, 1H), 3.90−3.77 (m, 2H), 3.76−3.67 (m, 1H), 3.65−3.49 (m, 4H), 3.32 (s, 3H), 2.47−2.39 (m, 2H), 2.36−2.30 (m, 1H), 2.10−1.94 (m, 4H), 1.72−1.63 (m, 1H), 1.55−1.39 (m, 3H), 1.35−1.28 (m, 1H) ppm. ESI-MS: m/z = 400 [M + H]+. (3S,6S)-3-(Methoxymethyl)-1,4-dioxo2,3,4,5,6,7,9,10,11,12,13,14-dodecahydro-1H-pyrrolo[2,1-i][1]oxa[4,7,10]triazacyclohexadecine-6-carboxylic Acid (22s). Obtained following the general procedure C from 21s. White solid (58 mg, 76%). 1H NMR (500 MHz, CDCl3) δ 7.40 (d, J = 8.0 Hz, 1H), 6.91−6.76 (m, 2H), 6.66 (d, J = 2.5 Hz, 1H), 6.17 (dd, J = 3.5, 2.5 Hz, 1H), 5.22−5.09 (m, 1H), 4.62 (dt, J = 8.0, 2.5 Hz, 1H), 4.59− 4.52 (m, 1H), 3.99 (dd, J = 10.0, 5.0 Hz, 1H), 3.93 (dd, J = 9.5, 2.0 Hz, 2H), 3.76−3.65 (m, 2H), 3.51−3.36 (m, 5H), 1.78−1.59 (m, 2H), 1.60−1.49 (m, 1H), 1.47−1.36 (m, 2H), 1.33−1.19 (m, 3H) ppm. ESI-MS: m/z = 382 [M + H]+. (13S,16S)-16-(Methoxymethyl)-15,18-dioxo5,6,7,8,9,10,13,14,15,16,17,18-dodecahydro-12H-imidazo[2,1i][1]oxa[4,7,10]triazacyclohexadecine-13-carboxylic Acid (22t). Obtained following the general procedure C from 21t. Brown solid (45 mg, 59%). 1H NMR (500 MHz, CDCl3) δ 8.13 (d, J = 6.7 Hz, 1H), 7.46 (d, J = 7.9 Hz, 1H), 7.10−7.04 (m, 2H), 4.68−4.62 (m, 1H), 4.56−4.50 (m, 1H), 4.03−3.94 (m, 2H), 3.80 (dd, J = 9.5, 3.0 Hz, 1H), 3.74 (dd, J = 10.0, 4.5 Hz, 1H), 3.63 (dd, J = 9.5, 3.0 Hz, 1H), 3.49−3.31 (m, 6H), 1.87−1.75 (m, 2H), 1.61−

1.45 (m, 2H), 1.43−1.32 (m, 2H), 1.30−1.21 (m, 2H) ppm. 13C NMR (100 MHz, DMSO) δ 171.02, 169.36, 159.53, 138.10, 127.89, 124.61, 70.37, 69.82, 69.32, 58.12, 52.40, 52.14, 45.47, 30.37, 28.91, 24.93, 24.23. ESI-MS: m/z = 383 [M + H]+. (14S,17S)-17-(Methoxymethyl)-16,19-dioxo6,7,8,9,10,11,14,15,16,17,18,19-dodecahydro-5H,13Himidazo[2,1-i][1]oxa[4,7,10]triazacycloheptadecine-14-carboxylic Acid (22u). Obtained following the general procedure C from 21u. Brown solid (57 mg, 72%); mp = 192−193 °C. 1H NMR (500 MHz, CDCl3) δ 8.22 (d, J = 6.5 Hz, 1H), 7.29 (d, J = 8.0 Hz, 1H), 7.09−6.99 (m, 2H), 5.06 (dt, J = 12.5, 5.5 Hz, 1H), 4.70−4.64 (m, 1H), 4.58 (dt, J = 6.0, 2.5 Hz, 1H), 3.99 (dd, J = 9.5, 5.0 Hz, 1H), 3.94−3.82 (m, 2H), 3.73 (dd, J = 9.5, 4.5 Hz, 1H), 3.67 (dd, J = 9.5, 3.5 Hz, 1H), 3.41 (s, 3H), 3.39−3.33 (m, 2H), 1.96−1.81 (m, 1H), 1.73−1.58 (m, 1H), 1.40−1.18 (m, 8H) ppm. 13C NMR (100 MHz, CDCl3) δ 172.76, 170.43, 159.91, 137.60, 127.84, 125.92, 71.33, 70.68, 69.56, 59.33, 53.75, 53.48, 49.00, 29.20, 28.30, 27.00, 24.28, 24.15. ESI-MS: m/z = 397 [M + H]+. (6S,9S)-6-(Methoxymethyl)-4,7-dioxo5,6,7,8,9,10,12,13,14,15,16,17-dodecahydro-4H-pyrazolo[5,1i][1]oxa[4,7,10]triazacyclohexadecine-9-carboxylic Acid (22v). Obtained following the general procedure C from 21v. White solid (57 mg, 75%). 1H NMR (500 MHz, CDCl3) δ 7.47 (d, J = 2.0 Hz, 1H), 6.60 (d, J = 2.0 Hz, 1H), 5.15−5.06 (m, 1H), 4.75−4.69 (m, 1H), 4.58−4.52 (m, 1H), 4.19 (dt, J = 13.5, 5.0 Hz, 1H), 3.90 (dd, J = 10.0, 5.5 Hz, 1H), 3.75 (dd, J = 10.0, 4.0 Hz, 1H), 3.70−3.62 (m, 2H), 3.48−3.28 (m, 5H), 2.01−1.90 (m, 1H), 1.76−1.66 (m, 1H), 1.51−1.29 (m, 3H), 1.28−0.99 (m, 3H) ppm. 13C NMR (100 MHz, DMSO) δ 171.18, 169.36, 159.65, 136.90, 134.37, 108.05, 71.09, 69.92, 69.17, 58.08, 52.53, 52.00, 50.80, 28.57, 26.80, 24.45, 24.39. ESI-MS: m/z = 383 [M + H]+. (6S,9S)-6-(Methoxymethyl)-4,7-dioxo5,6,7,8,9,10,13,14,15,16,17,18-dodecahydro-4H,12Hpyrazolo[5,1-i][1]oxa[4,7,10]triazacycloheptadecine-9-carboxylic Acid (22w). Obtained following the general procedure C from 21w. White solid (59 mg, 74%); mp = 239−240 °C. 1H NMR (500 MHz, DMSO) δ 8.64 (d, J = 8.5 Hz, 1H), 8.20 (d, J = 8.0 Hz, 1H), 7.46 (d, J = 2.0 Hz, 1H), 6.95 (d, J = 2.0 Hz, 1H), 4.90−4.76 (m, 2H), 4.52 (td, J = 7.5, 3.5 Hz, 1H), 4.11 (dt, J = 13.1, 7.3 Hz, 1H), 3.70−3.57 (m, 4H), 3.37−3.34 (m, 2H), 3.29 (s, 3H), 1.67−1.58 (m, 2H), 1.43−1.33 (m, 2H), 1.31−1.20 (m, 4H), 1.18−1.07 (m, 2H) ppm. ESI-MS: m/z = 397 [M + H]+. (6S,9S)-6-(Methoxymethyl)-1-methyl-4,7-dioxo4,5,6,7,8,9,10,12,13,14,15,16,17,18-tetradecahydro-1Hpyrazolo[4,3-i][1]oxa[4,7]diazacycloheptadecine-9-carboxylic Acid (22x). Obtained following the general procedure C from 21x. White solid (71 mg, 87%). 1H NMR (500 MHz, CDCl3) δ 7.74 (s, 1H), 7.55 (d, J = 6.5 Hz, 1H), 6.82 (s, 1H), 4.82−4.73 (m, 1H), 4.68−4.59 (m, 1H), 4.02−3.91 (m, 1H), 3.89−3.77 (m, 4H), 3.75− 3.65 (m, 2H), 3.52−3.30 (m, 6H), 2.64−2.53 (m, 1H), 1.77−1.62 (m, 1H), 1.61−1.46 (m, 2H), 1.44−1.18 (m, 8H) ppm. ESI-MS: m/z = 411 [M + H]+. General Procedure for Compounds 22y,22z Synthesis. A solution of the sulfur-containing diolefin (0.20 mmol, 1.0 equiv) in toluene (0.01 molar) was degassed with dry nitrogen for 15 min. The mixture was stirred for 5 min at 100 °C. after which, a degassed solution of grubbs second generation catalyst (0.10 equiv) in toluene (0.01 molar) was injected with a syringe for 30 min. The reaction was stirred for half an additional hour. After cooling to room temperature, the solvent was evaporated under reduced pressure and purified by flash chromatography on silica gel. The resulting products (a mixture of geometric isomers) were redissolved in methanol (0.05 molar) and stirred overnight with 10% Pd/C (0.5 equiv). The mixture was then filtered over Celite and concentrated to give the corresponding saturated macrocyclic ester (25a and 25b). The saturated macrocyclic ester (0.14 mmol, 1.0 equiv) in acetone (0.68 mL) was added aqueous LiOH 0.3 M (1.5 equiv) dropwise. The solution was stirred at room temperature for 6 h. The reaction mixture was then acidified to pH 2 with 3 N HCl and extracted with EtOAc, washed with brine, dried (Na2SO4), filtered, and concentrated to yield the crude 9196



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7.5, 4.5 Hz, 1H), 4.31−4.27 (m, 1H), 4.15−4.12 (m, 1H), 4.04 (dd, J = 9.5, 2.0 Hz, 1H), 3.64 (dd, J = 9.5, 4.0 Hz, 1H), 3.60 (dd, J = 9.5, 5.0 Hz, 1H), 3.50 (dd, J = 10.0, 4.0 Hz, 1H), 3.41 (s, 3H), 3.35−3.28 (m, 3H), 3.04 (dd, J = 14.0, 4.5 Hz, 1H), 2.83 (d, J = 5.0 Hz, 1H), 2.70 (dd, J = 14.0, 9.0 Hz, 1H), 1.99−1.91 (m, 1), 1.88−1.80 (m, 1H), 1.64−1.56 (m, 2H), 1.51−1.45 (m, 2H), 1.44−1.40 (m, 5H) ppm. 13C NMR (125 MHz, CDCl3) δ 207.19, 170.62, 169.58, 166.64, 157.44, 136.47, 133.72, 132.66, 129.43, 128.28, 126.68, 121.31, 120.39, 111.90, 71.56, 70.52, 69.00, 68.53, 59.38, 59.26, 55.24, 52.70, 52.49, 52.43, 37.03, 29.20, 28.59, 27.08, 26.93, 16.57. ESI-MS: m/z = 596 [M + H]+. HRMS m/z calcd for C32H41N3O8 [M + H]+ 596.2966, found 596.2965. (11S,14S)-14-(Methoxymethyl)-N-((S)-1-((R)-2-methyloxiran-2-yl)-1-oxo-3-phenylpropan-2-yl)-13,16-dioxo3,4,5,6,7,8,11,12,13,14,15,16-dodecahydro-2H,10H-benzo[i][1,11]dioxa[4,7]diazacyclooctadecine-11-carboxamide (24d). Obtained following the general procedure D from 22d. White solid (46 mg, 71%); mp = 157−158 °C; [α]20D = 353.3 (c 0.01, CHCl3). 1 H NMR (500 MHz, CDCl3) δ 8.89 (d, J = 5.5 Hz, 1H), 8.19 (dd, J = 8.0, 1.5 Hz, 1H), 7.49−7.43 (m, 1H), 7.21 (d, J = 8.0 Hz, 1H), 7.15− 6.98 (m, 7H), 6.78 (d, J = 8.0 Hz, 1H), 4.82 (td, J = 8.5, 5.5 Hz, 1H), 4.72−4.66 (m, 2H), 4.35−4.27 (m, 1H), 4.11−4.05 (m, 1H), 3.98 (dd, J = 9.5, 3.5 Hz, 1H), 3.72−3.61 (m, 2H), 3.52 (dd, J = 10.0, 4.0 Hz, 1H), 3.47−3.39 (m, 4H), 3.36−3.31 (m, 2H), 3.08 (dd, J = 14.0, 5.5 Hz, 1H), 2.83 (d, J = 5.0 Hz, 1H), 2.78 (dd, J = 14.0, 9.0 Hz, 1H), 2.02−1.96 (m, 1H), 1.88−1.79 (m, 1H), 1.61−1.52 (m, 1H), 1.50− 1.31 (m, 10H) ppm. 13C NMR (125 MHz, CDCl3) δ 207.39, 170.26, 169.56, 166.53, 157.61, 136.33, 133.62, 132.51, 129.37, 128.33, 126.80, 121.37, 120.70, 112.69, 71.72, 69.77, 69.57, 69.23, 59.32, 59.27, 55.28, 52.53, 52.49, 52.39, 37.13, 28.39, 28.00, 27.07, 26.65, 25.61, 16.49. ESI-MS: m/z = 610 [M + H]+. (12S,15S)-15-(Methoxymethyl)-N-((S)-1-((R)-2-methyloxiran-2-yl)-1-oxo-3-phenylpropan-2-yl)-14,17-dioxo2,3,4,5,6,7,8,9,12,13,14,15,16,17-tetradecahydro-11H-benzo[i][1,11]dioxa[4,7]diazacyclononadecine-12-carboxamide (24e). Obtained following the general procedure D from 22e. White solid (46 mg, 71%); mp = 148−150 °C, [α]20D = 178.3 (c 0.06, CHCl3). 1H NMR (500 MHz, CDCl3) δ 9.00 (d, J = 4.0 Hz, 1H), 8.27 (dd, J = 8.0, 2.0 Hz, 1H), 7.52−7.45 (m, 1H), 7.42 (d, J = 8.0 Hz, 1H), 7.18−6.99 (m, 7H), 6.80 (d, J = 8.0 Hz, 1H), 4.82 (td, J = 8.5, 5.5 Hz, 1H), 4.67−4.60 (m, 2H), 4.28−4.17 (m, 2H), 3.95 (dd, J = 9.5, 4.5 Hz, 1H), 3.70 (dd, J = 9.5, 4.5 Hz, 1H), 3.63 (dd, J = 9.5, 5.0 Hz, 1H), 3.51 (dd, J = 9.5, 4.5 Hz, 1H), 3.46−3.28 (m, 6H), 3.11 (dd, J = 14.0, 5.5 Hz, 1H), 2.88 (dd, J = 14.0, 8.5 Hz, 1H), 2.83 (d, J = 5.0 Hz, 1H), 2.03−1.97 (m, 1H), 1.95−1.77 (m, 2H), 1.66−1.53 (m, 2H), 1.49−1.29 (m, 10H) ppm. 13C NMR (125 MHz, CDCl3) δ 207.41, 170.00, 169.76, 167.14, 157.80, 136.62, 133.94, 132.83, 129.47, 128.31, 126.74, 121.23, 119.71, 112.00, 71.52, 69.81, 68.79, 68.30, 59.39, 59.14, 55.93, 52.81, 52.57, 52.42, 37.11, 28.71, 28.67, 28.18, 27.76, 25.10, 25.05, 16.52. ESI-MS: m/z = 624 [M + H]+. (13S,16S)-16-(Methoxymethyl)-N-((S)-1-((R)-2-methyloxiran-2-yl)-1-oxo-3-phenylpropan-2-yl)-15,18-dioxo3,4,5,6,7,8,9,10,13,14,15,16,17,18-tetradecahydro-2H,12Hbenzo[i][1,11]dioxa[4,7]diazacycloicosine-13-carboxamide (24f). Obtained following the general procedure D from 22f. White solid (42 mg, 66%); mp = 109−110 °C, [α]20D = −186.0 (c 0.044, CHCl3). 1H NMR (500 MHz, CDCl3) δ 9.02 (d, J = 4.5 Hz, 1H), 8.22 (dd, J = 8.0, 2.0 Hz, 1H), 7.49−7.41 (m, 2H), 7.25−7.11 (m, 5H), 7.06−6.97 (m, 3H), 4.83 (td, J = 8.0, 5.5 Hz, 1H), 4.70−4.64 (m, 1H), 4.58−4.50 (m, 1H), 4.24−4.06 (m, 2H), 3.89 (dd, J = 9.5, 4.5 Hz, 1H), 3.71 (dd, J = 9.5, 5.0 Hz, 1H), 3.59 (dd, J = 9.5, 6.5 Hz, 1H), 3.50−3.34 (m, 7H), 3.13 (dd, J = 14.0, 5.0 Hz, 1H), 2.91−2.82 (m, 2H), 2.12−2.02 (m, 1H), 1.95−1.85 (m, 1H), 1.64−1.23 (m, 15H) ppm. 13C NMR (125 MHz, CDCl3) δ 207.33, 170.15, 170.00, 166.20, 157.79, 136.30, 133.52, 132.62, 129.46, 128.49, 126.97, 121.11, 120.41, 112.24, 71.76, 70.79, 69.74, 68.69, 59.41, 59.16, 54.69, 52.81, 52.69, 52.54, 37.18, 29.06, 28.64, 28.46, 27.91, 27.85, 26.82, 25.28, 16.61. ESI-MS: m/z = 638 [M + H]+. (10S,13S)-19-Fluoro-13-(methoxymethyl)-N-((S)-1-((R)-2methyloxiran-2-yl)-1-oxo-3-phenylpropan-2-yl)-12,15-dioxo2,3,4,5,6,7,10,11,12,13,14,15-dodecahydro-9H-benzo[i][1,11]-

macrocyclic acid 22y and 22z (0.10 mmol, 1.0 equiv) for the next step without further purification. (9S,12S)-12-(Methoxymethyl)-11,14-dioxo3,4,5,6,7,8,9,10,11,12,13,14-dodecahydro-2H-benzo[b][1]oxa[5,8]diazacyclohexadecine-9-carboxylic Acid (22za). Obtained following the general procedure C from 21za. White solid (62 mg, 79%). 1H NMR (500 MHz, CDCl3) δ 9.02 (d, J = 8.5 Hz, 1H), 8.24 (dd, J = 8.0, 1.5 Hz, 1H), 7.51−7.44 (m, 1H), 7.11−7.05 (m, 1H), 7.00−6.92 (m, 2H), 4.97−4.91 (m, 1H), 4.66−4.59 (m, 1H), 4.25−4.19 (m, 1H), 4.18−4.08 (m, 2H), 3.52 (dd, J = 9.0, 4.0 Hz, 1H), 3.41 (s, 3H), 2.11−1.89 (m, 3H), 1.83−1.74 (m, 1H), 1.69−1.62 (m, 1H), 1.60− 1.52 (m, 1H), 1.50−1.29 (m, 6H) ppm. ESI-MS: m/z = 393 [M + H]+. (10S,13S)-13-(Methoxymethyl)-12,15-dioxo2,3,4,5,6,7,8,9,10,11,12,13,14,15-tetradecahydrobenzo[b][1]oxa[5,8]diazacycloheptadecine-10-carboxylic Acid (22zb). Obtained following the general procedure C from 21zb. White solid (66 mg, 81%). 1H NMR (500 MHz, CDCl3) δ 8.70 (d, J = 8.0 Hz, 1H), 8.13 (dd, J = 8.0, 1.5 Hz, 1H), 7.45−7.38 (m, 1H), 7.06−7.00 (m, 1H), 6.94 (d, J = 8.0 Hz, 1H), 6.89 (d, J = 8.0 Hz, 1H), 4.94−4.85 (m, 1H), 4.64 (td, J = 8.5, 3.5 Hz, 1H), 4.25−4.18 (m, 1H), 4.06−3.98 (m, 2H), 3.46 (dd, J = 9.0, 5.5 Hz, 1H), 3.35 (s, 3H), 2.02−1.91 (m, 2H), 1.71−1.44 (m, 4H), 1.41−1.11 (m, 8H) ppm. ESI-MS: m/z = 407 [M + H]+. 23 was synthesized following the procedure previously reported in our group.39 (8S,11S)-11-(Methoxymethyl)-N-((S)-1-((R)-2-methyloxiran2-yl)-1-oxo-3-phenylpropan-2-yl)-10,13-dioxo2,3,4,5,8,9,10,11,12,13-decahydro-7H-benzo[i][1,11]dioxa[4,7]diazacyclopentadecine-8-carboxamide (24a). Obtained following the general procedure D from 22a. White solid (42 mg, 74%); mp = 167−168 °C, [α]20D = 85.1 (c 0.04, CHCl3). 1H NMR (500 MHz, CDCl3) δ 8.03−7.97 (m, 2H), 7.49 (ddd, J = 8.5, 7.5, 2.0 Hz, 1H), 7.15−7.10 (m, 2H), 7.07−6.94 (m, 7H), 4.84−4.78 (m, 1H), 4.78−4.72 (m, 1H), 4.56−4.51 (m, 1H), 4.20−4.15 (m, 1H), 4.12−4.06 (m, 1H), 4.04 (dd, J = 9.0, 2.5 Hz, 1H), 3.76 (dd, J = 9.5, 4.5 Hz, 1H), 3.56 (dd, J = 9.5, 3.5 Hz, 1H), 3.53−3.44 (m, 2H), 3.37−3.30 (m, 5H), 3.06 (dd, J = 14.0, 4.0 Hz, 1H), 2.87 (d, J = 5.0 Hz, 1H), 2.70 (dd, J = 14.0, 9.5 Hz, 1H), 1.99−1.81 (m, 2H), 1.71− 1.60 (m, 1H), 1.51−1.41 (m, 4H) ppm. 13C NMR (125 MHz, CDCl3) δ 207.08, 170.74, 169.71, 167.01, 156.83, 136.37, 133.06, 131.68, 129.30, 128.32, 126.67, 123.37, 121.97, 113.88, 71.58, 70.01, 69.85, 69.40, 59.45, 59.39, 54.64, 53.11, 53.10, 52.55, 36.70, 26.55, 25.91, 16.68. ESI-MS: m/z = 568 [M + H]+. (9S,12S)-12-(Methoxymethyl)-N-((S)-1-((R)-2-methyloxiran2-yl)-1-oxo-3-phenylpropan-2-yl)-11,14-dioxo3,4,5,6,9,10,11,12,13,14-decahydro-2H,8H-benzo[i][1,11]dioxa[4,7]diazacyclohexadecine-9-carboxamide (24b). Obtained following the general procedure D from 22b. White solid (45 mg, 79%); mp = 171−173 °C, [α]20D = 229.5 (c 0.026, CHCl3). 1 H NMR (500 MHz, CDCl3) δ 8.78 (d, J = 5.0 Hz, 1H), 8.19 (d, J = 7.5 Hz, 1H), 7.54−7.44 (m, 1H), 7.10−6.92 (m, 9H), 4.80−4.67 (m, 2H), 4.66−4.58 (m, 1H), 4.39−4.20 (m, 2H), 4.07−3.96 (m, 2H), 3.68−3.58 (m, 1H), 3.56−3.50 (m, 1H), 3.40 (s, 3H), 3.34 (d, J = 5.0 Hz, 1H), 3.31−3.27 (m, 1H), 3.22−3.15 (m, 1H), 3.04 (dd, J = 13.5, 4.5 Hz, 1H), 2.82 (d, J = 4.0 Hz, 1H), 2.73 (dd, J = 13.5, 9.0 Hz, 1H), 1.93−1.87 (m, 1H), 1.83−1.76 (m, 1H), 1.65−1.54 (m, 2H), 1.48− 1.39 (m, 5H) ppm. 13C NMR (125 MHz, CDCl3) δ 207.68, 170.66, 169.59, 166.87, 157.15, 136.27, 133.73, 132.60, 129.29, 128.18, 126.59, 121.62, 120.96, 112.67, 71.57, 70.37, 69.76, 68.60, 59.45, 59.31, 55.49, 52.88, 52.65, 52.50, 36.79, 28.23, 28.19, 23.74, 16.54. ESI-MS: m/z = 582 [M + H]+. (10S,13S)-13-(Methoxymethyl)-N-((S)-1-((R)-2-methyloxiran-2-yl)-1-oxo-3-phenylpropan-2-yl)-12,15-dioxo2,3,4,5,6,7,10,11,12,13,14,15-dodecahydro-9H-benzo[i][1,11]dioxa[4,7]diazacycloheptadecine-10-carboxamide (24c). Obtained following the general procedure D from 22c. White solid (42 mg, 71%); mp = 69−71 °C, [α]20D = −23.8 (c 0.044, CHCl3). 1H NMR (500 MHz, CDCl3) δ 8.90 (d, J = 6.5 Hz, 1H), 8.23 (dd, J = 8.0, 2.0 Hz, 1H), 7.52−7.46 (m, 1H), 7.13−6.96 (m, 8H), 6.87 (d, J = 8.0 Hz, 1H), 4.78−4.72 (m, 1H), 4.70−4.65 (m, 1H), 4.58 (dt, J = 9197



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ppm. 13C NMR (125 MHz, CDCl3) δ 207.22, 170.87, 169.60, 166.46, 164.10, 158.81, 136.49, 134.26, 129.44, 128.29, 126.68, 113.42, 105.36, 99.14, 71.58, 70.46, 69.07, 68.47, 59.36, 59.22, 55.71, 55.04, 52.69, 52.42, 37.03, 29.08, 28.62, 27.11, 26.86, 16.54. ESI-MS: m/z = 626 [M + H]+. (10S,13S)-13-(Methoxymethyl)-18-methyl-N-((S)-1-((R)-2methyloxiran-2-yl)-1-oxo-3-phenylpropan-2-yl)-12,15-dioxo2,3,4,5,6,7,10,11,12,13,14,15-dodecahydro-9H-benzo[i][1,11]dioxa[4,7]diazacycloheptadecine-10-carboxamide (24k). Obtained following the general procedure D from 22k. White solid (50 mg, 80%); mp = 57−58 °C, [α]20D = 23.6 (c 0.022, CHCl3). 1H NMR (500 MHz, CDCl3) δ 8.84 (d, J = 6.5 Hz, 1H), 8.12 (d, J = 8.0 Hz, 1H), 7.18 (d, J = 8.1 Hz, 1H), 7.11−7.01 (m, 5H), 6.87 (d, J = 8.0 Hz, 1H), 6.83−6.76 (m, 2H), 4.79−4.69 (m, 2H), 4.58 (dt, J = 8.0, 5.5 Hz, 1H), 4.31−4.22 (m, 1H), 4.17−4.09 (m, 1H), 4.06 (dd, J = 9.5, 2.0 Hz, 1H), 3.61 (dd, J = 9.0, 3.5 Hz, 1H), 3.57 (dd, J = 9.5, 5.5 Hz, 1H), 3.49 (dd, J = 9.5, 4.0 Hz, 1H), 3.39 (s, 3H), 3.36−3.28 (m, 3H), 3.04 (dd, J = 14.0, 5.0 Hz, 1H), 2.82 (d, J = 5.0 Hz, 1H), 2.75 (dd, J = 14.0, 9.0 Hz, 1H), 2.39 (s, 3H), 2.00−1.91 (m, 1H), 1.88− 1.79 (m, 1H), 1.67−1.56 (m, 2H), 1.53−1.40 (m, 7H) ppm. 13C NMR (125 MHz, CDCl3) δ 207.25, 170.77, 169.59, 166.73, 157.39, 144.69, 136.47, 132.52, 129.43, 128.26, 126.65, 122.10, 117.62, 112.56, 71.54, 70.45, 68.86, 68.47, 59.36, 59.22, 55.12, 52.68, 52.45, 52.41, 37.00, 29.18, 28.58, 27.07, 26.85, 21.95, 16.53. ESI-MS: m/z = 610 [M + H]+. (10S,13S)-18-Chloro-13-(methoxymethyl)-N-((S)-1-((R)-2methyloxiran-2-yl)-1-oxo-3-phenylpropan-2-yl)-12,15-dioxo2,3,4,5,6,7,10,11,12,13,14,15-dodecahydro-9H-benzo[i][1,11]dioxa[4,7]diazacycloheptadecine-10-carboxamide (24l). Obtained following the general procedure D from 22l. White solid (44 mg, 70%); mp = 69−70 °C, [α]20D = 473.5 (c 0.012, CHCl3). 1H NMR (500 MHz, CDCl3) δ 8.73 (d, J = 7.0 Hz, 1H), 8.16 (d, J = 8.5 Hz, 1H), 7.12−7.07 (m, 4H), 7.06−7.02 (m, 3H), 6.99 (d, J = 1.5 Hz, 1H), 6.80 (d, J = 8.0 Hz, 1H), 4.78−4.68 (m, 2H), 4.58 (dt, J = 7.5, 5.5 Hz, 1H), 4.30−4.24 (m, 1H), 4.15−4.08 (m, 1H), 4.05 (dd, J = 9.0, 2.0 Hz, 1H), 3.64−3.57 (m, 2H), 3.51 (dd, J = 9.5, 4.0 Hz, 1H), 3.39 (s, 3H), 3.38−3.29 (m, 3H), 3.04 (dd, J = 14.0, 5.0 Hz, 1H), 2.82 (d, J = 5.0 Hz, 1H), 2.75 (dd, J = 14.0, 8.5 Hz, 1H), 2.00−1.91 (m, 1H), 1.89−1.81 (m, 1H), 1.67−1.57 (m, 2H), 1.54−1.40 (m, 7H) ppm. 13C NMR (125 MHz, CDCl3) δ 207.02, 170.42, 169.46, 165.17, 155.90, 136.24, 133.18, 132.21, 129.29, 128.29, 126.72, 126.63, 121.66, 113.37, 71.47, 70.55, 69.55, 68.56, 59.35, 59.26, 55.08, 52.69, 52.48, 52.44, 36.90, 29.12, 28.60, 27.04, 26.93, 16.58. ESI-MS: m/z = 630 [M + H]+. (10S,13S)-17-Chloro-13-(methoxymethyl)-N-((S)-1-((R)-2methyloxiran-2-yl)-1-oxo-3-phenylpropan-2-yl)-12,15-dioxo2,3,4,5,6,7,10,11,12,13,14,15-dodecahydro-9H-benzo[i][1,11]dioxa[4,7]diazacycloheptadecine-10-carboxamide (24m). Obtained following the general procedure D from 22m. White solid (49 mg, 78%); mp = 69−70 °C, [α]20D = 391.7 (c 0.016, CHCl3). 1H NMR (500 MHz, CDCl3) δ 8.82 (d, J = 6.5 Hz, 1H), 8.19 (d, J = 3.0 Hz, 1H), 7.41 (dd, J = 9.0, 2.5 Hz, 1H), 7.14−7.02 (m, 6H), 6.93 (d, J = 9.0 Hz, 1H), 6.78 (d, J = 8.0 Hz, 1H), 4.76 (td, J = 8.5, 5.0 Hz, 1H), 4.73−4.69 (m, 1H), 4.59 (dt, J = 7.5, 5.0 Hz, 1H), 4.29−4.22 (m, 1H), 4.13−4.03 (m, 2H), 3.63−3.57 (m, 2H), 3.52 (dd, J = 10.0, 4.0 Hz, 1H), 3.42−3.29 (m, 6H), 3.06 (dd, J = 14.0, 4.5 Hz, 1H), 2.83 (d, J = 5.0 Hz, 1H), 2.79−2.74 (m, 1H), 2.01−1.90 (m, 1H), 1.88−1.80 (m, 1H), 1.66−1.56 (m, 2H), 1.55−1.39 (m, 7H) ppm. 13C NMR (125 MHz, CDCl3) δ 207.15, 170.50, 169.48, 165.60, 157.72, 139.38, 136.27, 133.65, 129.30, 128.27, 126.72, 121.56, 118.90, 112.63, 71.45, 70.51, 69.62, 68.50, 59.32, 59.24, 55.03, 52.63, 52.46, 52.40, 36.89, 29.02, 28.54, 27.01, 26.86, 16.52. ESI-MS: m/z = 630 [M + H]+. (14S,17S)-17-(Methoxymethyl)-N-((S)-1-((R)-2-methyloxiran-2-yl)-1-oxo-3-phenylpropan-2-yl)-16,19-dioxo6,7,8,9,10,11,14,15,16,17,18,19-dodecahydro-13H-pyrido[2,3i][1,11]dioxa[4,7]diazacycloheptadecine-14-carboxamide (24n). Obtained following the general procedure D from 22n. Viscous liquid (39 mg, 65%), [α]20D = 233.1 (c 0.026, CHCl3). 1H NMR (500 MHz, CDCl3) δ 8.64 (d, J = 8.0 Hz, 1H), 8.38 (d, J = 4.0 Hz, 1H), 7.36−7.28 (m, 1H), 7.24−7.16 (m, 1H), 7.06−6.90 (m, 5H), 5.06−4.95 (m, 1H), 4.84 (td, J = 8.5, 5.5 Hz, 1H), 4.80−4.72

dioxa[4,7]diazacycloheptadecine-10-carboxamide (24g). Obtained following the general procedure D from 22g. White solid (43 mg, 70%); mp = 169−171 °C, [α]20D = 81.5 (c 0.068, CHCl3). 1H NMR (500 MHz, CDCl3) δ 8.84 (d, J = 6.0 Hz, 1H), 8.01−7.95 (m, 1H), 7.32−7.20 (m, 3H), 7.12−7.01 (m, 6H), 6.85 (d, J = 8.5 Hz, 1H), 4.80 (td, J = 9.0, 5.0 Hz, 1H), 4.67−4.57 (m, 2H), 4.56−4.49 (m, 1H), 4.41−4.33 (m, 1H), 4.03 (dd, J = 9.5, 2.5 Hz, 1H), 3.70− 3.61 (m, 2H), 3.52 (dd, J = 9.5, 4.0 Hz, 1H), 3.44−3.38 (m, 4H), 3.35 (d, J = 5.0 Hz, 1H), 3.33−3.26 (m, 1H), 3.08 (dd, J = 14.0, 5.0 Hz, 1H), 2.87−2.77 (m, 2H), 2.03−1.92 (m, 1H), 1.91−1.82 (m, 1H), 1.82−1.72 (m, 1H), 1.63−1.48 (m, 2H), 1.47−1.37 (m, 6H) ppm. 13 C NMR (125 MHz, CDCl3) δ 207.22, 169.98, 169.71, 165.90, 154.11 (d, 1JC−F = 246.4 Hz), 145.36 (d, 2JC−F = 10.8 Hz), 136.57, 129.35, 128.30, 127.45 (d, 3JC−F = 2.7 Hz), 126.72, 125.15, 122.85 (d, 3 JC−F = 7.9 Hz), 121.05 (d, 2JC−F= 20.3 Hz), 75.27, 71.46, 70.82, 68.88, 59.37, 59.28, 55.61, 52.74, 52.69, 52.44, 36.86, 30.07, 28.25, 27.21, 26.04, 16.58. ESI-MS: m/z = 614 [M + H]+. (10S,13S)-19-Methoxy-13-(methoxymethyl)-N-((S)-1-((R)-2methyloxiran-2-yl)-1-oxo-3-phenylpropan-2-yl)-12,15-dioxo2,3,4,5,6,7,10,11,12,13,14,15-dodecahydro-9H-benzo[i][1,11]dioxa[4,7]diazacycloheptadecine-10-carboxamide (24h). Obtained following the general procedure D from 22h. White solid (45 mg, 72%); mp = 60−61 °C, [α]20D = 141.8 (c 0.044, CHCl3). 1H NMR (500 MHz, CDCl3) δ 8.99 (d, J = 5.5 Hz, 1H), 7.78 (dd, J = 7.5, 2.0 Hz, 1H), 7.30−7.27 (m, 1H), 7.13−7.04 (m, 7H), 6.86 (d, J = 8.5 Hz, 1H), 4.81 (td, J = 8.5, 5.0 Hz, 1H), 4.65−4.58 (m, 2H), 4.30 (dt, J = 10.0, 6.5 Hz, 1H), 4.13 (dt, J = 10.0, 6.5 Hz, 1H), 4.00 (dd, J = 9.5, 3.0 Hz, 1H), 3.91 (s, 3H), 3.72 (dd, J = 9.5, 3.5 Hz, 1H), 3.63 (dd, J = 9.5, 4.0 Hz, 1H), 3.49 (dd, J = 9.5, 4.0 Hz, 1H), 3.44−3.35 (m, 5H), 3.28−3.22 (m, 1H), 3.08 (dd, J = 14.0, 5.0 Hz, 1H), 2.86− 2.80 (m, 2H), 1.86−1.73 (m, 2H), 1.57−1.50 (m, 1H), 1.44 (s, 3H), 1.41−1.30 (m, 5H) ppm. 13C NMR (125 MHz, CDCl3) δ 207.21, 170.06, 169.84, 166.96, 152.38, 147.19, 136.69, 129.35, 128.28, 126.63, 125.34, 123.86, 123.47, 116.49, 74.87, 71.58, 70.85, 69.10, 59.37, 59.24, 56.37, 55.83, 52.81, 52.69, 52.42, 36.81, 29.67, 28.14, 26.85, 25.18, 16.59. ESI-MS: m/z = 626 [M + H]+. (10S,13S)-18-Fluoro-13-(methoxymethyl)-N-((S)-1-((R)-2methyloxiran-2-yl)-1-oxo-3-phenylpropan-2-yl)-12,15-dioxo2,3,4,5,6,7,10,11,12,13,14,15-dodecahydro-9H-benzo[i][1,11]dioxa[4,7]diazacycloheptadecine-10-carboxamide (24i). Obtained following the general procedure D from 22i. White solid (42 mg, 68%); mp = 52−54 °C, [α]20D = 40.1 (c 0.03, CHCl3). 1H NMR (500 MHz, CDCl3) δ 8.72 (d, J = 6.5 Hz, 1H), 8.28−8.21 (m, 1H), 7.12−7.02 (m, 6H), 6.84−6.74 (m, 2H), 6.71 (dd, J = 10.5, 2.5 Hz, 1H), 4.79−4.70 (m, 2H), 4.59 (dt, J = 8.0, 5.5 Hz, 1H), 4.30−4.22 (m, 1H), 4.13−4.03 (m, 2H), 3.64−3.56 (m, 2H), 3.51 (dd, J = 9.9, 4.0 Hz, 1H), 3.40 (s, 3H), 3.37−3.31 (m, 3H), 3.05 (dd, J = 14.0, 5.0 Hz, 1H), 2.83 (d, J = 5.0 Hz, 1H), 2.75 (dd, J = 14.0, 9.0 Hz, 1H), 2.02−1.93 (m, 1H), 1.90−1.81 (m, 1H), 1.69−1.58 (m, 2H), 1.57− 1.40 (m, 7H) ppm. 13C NMR (125 MHz, CDCl3) δ 207.18, 170.60, 169.51, 165.95 (d, 1JC−F = 253.5 Hz), 165.65, 158.83 (d, 3JC−F = 10.6 Hz), 136.34, 134.62 (d, 3JC−F = 10.8 Hz), 129.35, 128.29, 126.74, 116.72 (d, 4JC−F = 3.0 Hz), 108.30 (d, 2JC−F = 21.3 Hz), 100.13 (d, 2 JC−F = 26.3 Hz), 71.51, 70.52, 69.67, 68.51, 59.35, 59.26, 55.03, 52.68, 52.45, 52.43, 36.96, 29.00, 28.59, 27.05, 26.89, 16.55. ESI-MS: m/z = 614 [M + H]+. (10S,13S)-18-Methoxy-13-(methoxymethyl)-N-((S)-1-((R)-2methyloxiran-2-yl)-1-oxo-3-phenylpropan-2-yl)-12,15-dioxo2,3,4,5,6,7,10,11,12,13,14,15-dodecahydro-9H-benzo[i][1,11]dioxa[4,7]diazacycloheptadecine-10-carboxamide (24j). Obtained following the general procedure D from 22j. White solid (50 mg, 80%); mp = 60−61 °C, [α]20D = 33.5 (c 0.02, CHCl3). 1H NMR (500 MHz, CDCl3) δ 8.74 (d, J = 6.5 Hz, 1H), 8.19 (d, J = 9.0 Hz, 1H), 7.18 (d, J = 8.0 Hz, 1H), 7.11−7.03 (m, 5H), 6.82 (d, J = 8.0 Hz, 1H), 6.57 (dd, J = 8.8, 2.5 Hz, 1H), 6.50 (d, J = 2.5 Hz, 1H), 4.78−4.70 (m, 2H), 4.58 (dt, J = 8.0, 5.5 Hz, 1H), 4.28−4.21 (m, 1H), 4.13−4.02 (m, 2H), 3.85 (s, 3H), 3.63−3.55 (m, 2H), 3.49 (dd, J = 10.0, 4.0 Hz, 1H), 3.39 (s, 3H), 3.36−3.30 (m, 3H), 3.04 (dd, J = 14.0, 5.0 Hz, 1H), 2.82 (d, J = 5.0 Hz, 1H), 2.76 (dd, J = 14.0, 9.0 Hz, 1H), 2.00−1.91 (m, 1H), 1.88−1.79 (m, 1H), 1.65−1.37 (m, 9H) 9198



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3.90 (dd, J = 10.5, 2.5 Hz, 1H), 3.75−3.69 (m, 1H), 3.67 (dd, J = 9.0, 4.0 Hz, 1H), 3.62 (dd, J = 10.0, 4.0 Hz, 1H), 3.60−3.55 (m, 1H), 3.55−3.48 (m, 2H), 3.45−3.39 (m, 1H), 3.32−3.26 (m, 4H), 3.04 (dd, J = 14.0, 5.0 Hz, 1H), 2.86−2.78 (m, 2H), 2.41−2.32 (m, 1H), 2.12−1.87 (m, 5H), 1.71−1.61 (m, 1H), 1.57−1.45 (m, 1H), 1.44− 1.31 (m, 5H), 1.31−1.24 (m, 2H). 13C NMR (125 MHz, CDCl3) δ 207.05, 173.70, 172.49, 170.46, 169.29, 136.87, 129.79, 128.36, 126.66, 71.09, 69.03, 67.48, 62.26, 59.39, 59.17, 54.74, 54.10, 52.36, 47.47, 37.56, 33.74, 29.75, 28.27, 25.32, 23.51, 21.04, 16.49. ESI-MS: m/z = 587 [M + H]+. (3S,6S)-3-(Methoxymethyl)-N-((S)-1-((R)-2-methyloxiran-2yl)-1-oxo-3-phenylpropan-2-yl)-1,4-dioxo2,3,4,5,6,7,9,10,11,12,13,14-dodecahydro-1H-pyrrolo [2,1-i][1]oxa[4,7,10]triazacyclohexadecine-6-carboxamide (24s). Obtained following the general procedure D from 22s. White solid (41 mg, 72%); mp = 167−168 °C, [α]20D = −320.8 (c 0.2, CHCl3). 1 H NMR (500 MHz, CDCl3) δ 7.85 (d, J = 8.5 Hz, 1H), 7.31−7.18 (m, 6H), 6.86−6.81 (m, 1H), 6.74 (d, J = 5.0 Hz, 1H), 6.62 (dd, J = 3.9, 1.4 Hz, 1H), 6.17−6.12 (m, 1H), 5.12−5.03 (m, 2H), 4.51 (q, J = 4.5 Hz, 1H), 4.45−4.40 (m, 1H), 4.03 (dd, J = 9.5, 4.5 Hz, 1H), 3.87−3.77 (m, 2H), 3.72 (dd, J = 9.5, 4.5 Hz, 1H), 3.54 (dd, J = 9.5, 3.5 Hz, 1H), 3.44−3.38 (m, 4H), 3.37−3.29 (m, 2H), 3.15 (dd, J = 14.0, 6.5 Hz, 1H), 2.98 (dd, J = 14.2, 7.8 Hz, 1H), 2.78 (d, J = 5.0 Hz, 1H), 1.66−1.57 (m, 1H), 1.55−1.41 (m, 3H), 1.37−1.29 (m, 5H), 1.24−1.11 (m, 2H) ppm. 13C NMR (125 MHz, CDCl3) δ 207.54, 170.42, 169.78, 163.78, 136.50, 129.12, 128.41, 127.52, 126.77, 124.88, 113.83, 108.49, 72.30, 70.60, 69.33, 59.16, 59.10, 54.93, 54.29, 52.13, 51.25, 46.10, 37.32, 31.57, 29.23, 26.81, 25.71, 16.27. ESI-MS: m/z = 569 [M + H]+. (13S,16S)-16-(Methoxymethyl)-N-((S)-1-((R)-2-methyloxiran-2-yl)-1-oxo-3-phenylpropan-2-yl)-15,18-dioxo5,6,7,8,9,10,13,14,15,16,17,18-dodecahydro-12H-imidazo[2,1i][1]oxa[4,7,10]triazacyclohexadecine-13-carboxamide (24t). Obtained following the general procedure D from 22t. White solid (41 mg, 72%); mp = 65−66 °C, [α]20D = 34.9 (c 0.02, CHCl3). 1H NMR (500 MHz, CDCl3) δ 8.13 (d, J = 5.0 Hz, 1H), 7.62 (d, J = 8.0 Hz, 1H), 7.31−7.15 (m, 6H), 7.10 (s, 1H), 7.07 (s, 1H), 5.40 (td, J = 13.0, 3.5 Hz, 1H), 5.08−5.01 (m, 1H), 4.53 (dd, J = 9.0, 4.5 Hz, 1H), 4.41−4.34 (m, 1H), 4.02 (dd, J = 9.5, 4.5 Hz, 1H), 3.94 (dt, J = 13.5, 3.5 Hz, 1H), 3.79 (dd, J = 9.5, 2.0 Hz, 1H), 3.74 (dd, J = 9.5, 4.0 Hz, 1H), 3.56 (dd, J = 9.5, 3.5 Hz, 1H), 3.40 (s, 3H), 3.38−3.32 (m, 2H), 3.29 (d, J = 5.0 Hz, 1H), 3.14 (dd, J = 14.0, 6.0 Hz, 1H), 2.96 (dd, J = 14.0, 7.5 Hz, 1H), 2.80 (d, J = 5.0 Hz, 1H), 1.76−1.66 (m, 1H), 1.66−1.56 (m, 1H), 1.48−1.38 (m, 2H), 1.36 (s, 3H), 1.33−1.26 (m, 1H), 1.17−1.09 (m, 1H), 1.08−1.00 (m, 1H), 0.98−0.89 (m, 1H) ppm. 13C NMR (125 MHz, CDCl3) δ 207.56, 170.31, 169.37, 161.22, 138.30, 136.33, 129.19, 129.12, 128.49, 126.88, 124.15, 71.86, 70.23, 69.20, 59.28, 59.12, 54.65, 54.52, 52.20, 51.28, 45.74, 37.44, 31.19, 29.97, 26.43, 25.49, 16.31. ESI-MS: m/z = 570 [M + H]+. HRMS m/z calcd for C29H39N5O7 [M + H]+ 570.2922, found 570.2928. (14S,17S)-17-(Methoxymethyl)-N-((S)-1-((R)-2-methyloxiran-2-yl)-1-oxo-3-phenylpropan-2-yl)-16,19-dioxo6,7,8,9,10,11,14,15,16,17,18,19-dodecahydro-5H,13Himidazo[2,1-i][1]oxa[4,7,10]triazacycloheptadecine-14-carboxamide (24u). Obtained following the general procedure D from 22u. White solid (52 mg, 88%); mp = 59−60 °C, [α]20D = 473.6 (c 0.034, CHCl3). 1H NMR (500 MHz, CDCl3) δ 8.16 (d, J = 6.0 Hz, 1H), 7.48 (d, J = 8.5 Hz, 1H), 7.31−7.26 (m, 2H), 7.23−7.18 (m, 3H), 7.13 (d, J = 8.0 Hz, 1H), 7.05 (d, J = 1.0 Hz, 1H), 7.01 (d, J = 1.0 Hz, 1H), 5.10 (dt, J = 13.0, 5.0 Hz, 1H), 5.05−4.99 (m, 1H), 4.57−4.52 (m, 1H), 4.37 (ddd, J = 7.5, 3.5, 2.0 Hz, 1H), 4.02 (dd, J = 9.5, 4.5 Hz, 1H), 3.85 (dd, J = 9.5, 2.0 Hz, 1H), 3.82−3.75 (m, 1H), 3.71 (dd, J = 9.5, 4.0 Hz, 1H), 3.50 (dd, J = 9.5, 3.5 Hz, 1H), 3.39 (s, 3H), 3.35−3.30 (m, 1H), 3.28−3.22 (m, 2H), 3.10 (dd, J = 14.0, 8.0 Hz, 1H), 2.96 (dd, J = 13.5, 6.5 Hz, 1H), 2.74 (d, J = 5.0 Hz, 1H), 1.99−1.88 (m, 1H), 1.50−1.42 (m, 1H), 1.36−1.12 (m, 11H) ppm. 13 C NMR (126 MHz, CDCl3) δ 207.87, 169.96, 169.23, 160.32, 137.07, 136.02, 129.34, 128.41, 127.93, 126.91, 126.46, 70.87, 70.50, 68.95, 59.22, 59.03, 54.53, 54.26, 51.95, 51.09, 49.18, 38.00, 28.65, 27.52, 26.48, 23.35, 18.43, 16.01. ESI-MS: m/z = 584 [M + H]+.

(m, 1H), 4.23−4.15 (m, 1H), 4.15−4.07 (m, 1H), 4.07−3.98 (m, 1H), 3.64 (dd, J = 10.5, 8.5 Hz, 1H), 3.58−3.50 (m, 2H), 3.49−3.43 (m, 1H), 3.40−3.34 (m, 4H), 3.32 (d, J = 5.0 Hz, 1H), 3.03−2.94 (m, 1H), 2.83 (d, J = 5.0 Hz, 1H), 2.75−2.65 (m, 1H), 2.01−1.89 (m, 1H), 1.85−1.72 (m, 2H), 1.66−1.58 (m, 2H), 1.57−1.45 (m, 3H), 1.44 (s, 3H) ppm. 13C NMR (125 MHz, CDCl3) δ 206.75, 170.61, 169.96, 164.16, 154.69, 141.39, 137.78, 136.52, 129.38, 128.14, 126.96, 126.58, 120.34, 71.78, 70.12, 69.38, 69.07, 59.33, 59.15, 54.20, 53.16, 52.45, 52.07, 36.93, 29.37, 29.17, 27.62, 27.42, 16.67. ESI-MS: m/z = 597 [M + H]+. (13S,16S)-16-(Methoxymethyl)-N-((S)-1-((R)-2-methyloxiran-2-yl)-1-oxo-3-phenylpropan-2-yl)-15,18-dioxo-2,11-dioxa14,17-diazabicyclo[17.3.1]-tricosa-1(23),19,21-triene-13-carboxamide (24o). Obtained following the general procedure D from 22o. Viscous liquid (50 mg, 83%); [α]20D = 66.8 (c 0.074, CHCl3). 1 H NMR (500 MHz, CDCl3) δ 7.44−7.38 (m, 2H), 7.32−7.29 (m, 1H), 7.25 (d, J = 8.0 Hz, 1H), 7.16−7.12 (m, 2H), 7.11−7.03 (m, 5H), 6.97 (d, J = 5.5 Hz, 1H), 4.82 (td, J = 8.5, 4.5 Hz, 1H), 4.73− 4.68 (m, 1H), 4.46−4.41 (m, 1H), 4.22−4.10 (m, 2H), 4.03 (dd, J = 10.0, 4.0 Hz, 1H), 3.81 (dd, J = 9.5, 2.5 Hz, 1H), 3.69 (dd, J = 9.5, 4.0 Hz, 1H), 3.45 (dd, J = 9.5, 4.0 Hz, 1H), 3.40 (s, 3H), 3.37 (d, J = 5.0 Hz, 1H), 3.20−3.15 (m, 2H), 3.11 (dd, J = 14.0, 4.5 Hz, 1H), 2.88− 2.81 (m, 2H), 1.91−1.76 (m, 3H), 1.62−1.53 (m, 1H), 1.52−1.40 (m, 5H), 1.39−1.22 (m, 6H) ppm. 13C NMR (125 MHz, CDCl3) δ 207.18, 170.11, 169.39, 169.11, 159.23, 136.74, 135.15, 130.35, 129.40, 128.32, 126.70, 119.80, 117.28, 114.78, 72.16, 70.83, 69.46, 68.50, 59.40, 59.30, 54.88, 53.42, 53.06, 52.47, 36.84, 29.23, 28.56, 27.49, 27.15, 25.89, 24.90, 16.63. ESI-MS: m/z = 624 [M + H]+. (6S,9S)-6-(Methoxymethyl)-1-methyl-N-((S)-1-((R)-2-methyloxiran-2-yl)-1-oxo-3-phenylpropan-2-yl)-4,7- dioxo1,4,5,6,7,8,9,10,12,13,14,15,16,17-tetradecahydropyrazolo[4,3-i][1,11]dioxa[4,7]diazacycloheptadecine-9-carboxamide (24p). Obtained following the general procedure D from 22p. White solid (50 mg, 83%); mp = 62−63 °C, [α]20D = 62.3 (c 0.04, CHCl3). 1 H NMR (500 MHz, CDCl3) δ 7.75 (s, 1H), 7.23−7.09 (m, 6H), 7.06 (d, J = 8.0 Hz, 1H), 6.90 (d, J = 6.0 Hz, 1H), 4.87 (td, J = 8.0, 5.5 Hz, 1H), 4.69−4.61 (m, 1H), 4.55−4.50 (m, 1H), 4.38−4.0 (m, 1H), 4.18 (ddd, J = 10.0, 8.0, 5.5 Hz, 1H), 4.00 (dd, J = 9.5, 3.5 Hz, 1H), 3.76−3.69 (m, 4H), 3.61 (dd, J = 9.5, 4.5 Hz, 1H), 3.54 (dd, J = 9.5, 3.5 Hz, 1H), 3.45−3.27 (m, 6H), 3.10 (dd, J = 14.0, 5.5 Hz, 1H), 2.90−2.81 (m, 2H), 1.90−1.81 (m, 1H), 1.79−1.71 (m, 1H), 1.49− 1.34 (m, 9H) ppm. 13C NMR (125 MHz, CDCl3) δ 207.38, 170.21, 169.60, 163.19, 153.09, 139.36, 136.21, 129.37, 128.48, 126.92, 102.43, 75.67, 71.27, 70.80, 69.26, 59.30, 59.28, 53.99, 53.29, 52.38, 37.17, 34.84, 28.62, 28.22, 25.34, 24.00, 16.51. HRMS m/z calcd for C30H41N5O8 [M + H]+ 600.3028, found 600.3017. (7S,10S)-7-(Methoxymethyl)-N-((S)-1-((R)-2-methyloxiran-2yl)-1-oxo-3-phenylpropan-2-yl)-5,8-dioxo6,7,8,9,10,11,13,14,15,16,17,18-dodecahydro-5H-pyrido[3,2i][1,11]dioxa[4,7]diazacycloheptadecine-10-carboxamide (24q). Obtained following the general procedure C from 22q. White solid (40 mg, 67%); mp = 63−64 °C, [α]20D = 172.6 (c 0.04, CHCl3). 1 H NMR (500 MHz, CDCl3) δ 8.90 (d, J = 7.0 Hz, 1H), 8.50 (dd, J = 7.6, 2.0 Hz, 1H), 8.29 (dd, J = 4.8, 2.0 Hz, 1H), 7.12−6.98 (m, 7H), 6.81 (d, J = 7.8 Hz, 1H), 4.78−4.69 (m, 3H), 4.60−4.53 (m, 1H), 4.41−4.35 (m, 1H), 4.07 (dd, J = 9.3, 2.1 Hz, 1H), 3.65−3.58 (m, 2H), 3.50 (dd, J = 9.8, 4.0 Hz, 1H), 3.41 (s, 3H), 3.37−3.27 (m, 3H), 3.04 (dd, J = 13.9, 4.9 Hz, 1H), 2.82 (d, J = 5.0 Hz, 1H), 2.75 (dd, J = 13.9, 8.6 Hz, 1H), 1.93−1.86 (m, 2H), 1.63−1.39 (m, 9H) ppm. 13C NMR (125 MHz, CDCl3) δ 207.05, 170.43, 169.44, 165.09, 160.91, 150.51, 141.80, 136.26, 129.30, 128.27, 126.77, 117.82, 115.00, 71.52, 70.68, 68.59, 67.92, 59.30, 59.29, 54.95, 52.67, 52.57, 52.42, 36.97, 28.94, 28.78, 27.24, 27.08, 16.54. ESI-MS: m/z = 597 [M + H]+. (3S,6S,18aS)-3-(Methoxymethyl)-N-((S)-1-((R)-2-methyloxiran-2-yl)-1-oxo-3-phenylpropan-2-yl)-1,4,14-trioxohexadecahydro-1H-pyrrolo[2,1-i][1]oxa[4,7,10] triazacyclohexadecine6-carboxamide (24r). Obtained following the general procedure D from 22r. Viscous liquid (45 mg, 77%); [α]20D = −197.5 (c 0.02, CHCl3). 1H NMR (500 MHz, CDCl3) δ 7.66 (d, J = 7.0 Hz, 1H), 7.33−7.13 (m, 6H), 6.56 (d, J = 7.0 Hz, 1H), 4.77 (td, J = 9.0, 5.0 Hz, 1H), 4.61−4.53 (m, 1H), 4.44−4.37 (m, 1H), 4.37−4.32 (m, 1H), 9199



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1.78 (m, 2H), 1.63−1.44 (m, 6H), 1.42 (s, 3H) ppm. 13C NMR (125 MHz, CDCl3) δ 207.20, 171.04, 169.37, 162.67, 157.08, 136.22, 130.44, 129.48, 128.42, 126.86, 115.74, 115.09, 72.49, 71.75, 70.57, 68.57, 59.35, 59.25, 53.97, 52.78, 52.44, 52.35, 37.20, 29.42, 29.24, 27.19, 26.94, 16.54. ESI-MS: m/z = 602 [M + H]+. (13S,16S)-16-(Methoxymethyl)-2-methyl-N-((S)-1-((R)-2methyloxiran-2-yl)-1-oxo-3-phenylpropan-2-yl)-15,18-dioxo5,6,7,8,9,10,13,14,15,16,17,18-dodecahydro-12H-thiazolo[5,4i][1,11]dioxa[4,7]diazacycloheptadecine-13-carboxamide (24z). Obtained following the general procedure D from 22z. White solid (46 mg, 75%); mp = 63−64 °C, [α]20D = 154.6 (c 0.016, CHCl3). 1H NMR (500 MHz, CDCl3) δ 7.96 (d, J = 7.5 Hz, 1H), 7.21−7.13 (m, 3H), 7.11−7.07 (m, 2H), 7.04 (d, J = 8.0 Hz, 1H), 6.86 (d, J = 7.5 Hz, 1H), 4.78−4.69 (m, 2H), 4.66−4.60 (m, 1H), 4.54 (dt, J = 9.0, 5.0 Hz, 1H), 4.45 (dt, J = 10.5, 4.0 Hz, 1H), 4.06 (dd, J = 9.0, 2.0 Hz, 1H), 3.71 (dd, J = 10.0, 5.0 Hz, 1H), 3.56 (dd, J = 9.0, 3.5 Hz, 1H), 3.47 (dd, J = 10.0, 4.0 Hz, 1H), 3.44−3.37 (m, 4H), 3.34−3.27 (m, 2H), 3.05 (dd, J = 14.0, 5.0 Hz, 1H), 2.83 (d, J = 5.0 Hz, 1H), 2.81−2.75 (m, 1H), 2.61 (s, 3H), 1.88−1.80 (m, 2H), 1.58−1.45 (m, 6H), 1.42 (s, 3H) ppm. 13C NMR (125 MHz, CDCl3) δ 207.12, 170.88, 169.38, 168.03, 161.84, 161.09, 136.26, 129.43, 128.38, 126.89, 105.79, 72.15, 71.66, 70.73, 68.69, 59.33, 59.28, 53.98, 52.76, 52.45, 37.11, 29.39, 29.34, 27.29, 27.11, 20.35, 16.54. ESI-MS: m/z = 617 [M + H]+. (9S,12S)-12-(Methoxymethyl)-N-((S)-1-((R)-2-methyloxiran2-yl)-1-oxo-3-phenylpropan-2-yl)-11,14-dioxo3,4,5,6,7,8,9,10,11,12,13,14-dodecahydro-2H-benzo[b][1]oxa[5,8]diazacyclohexadecine-9-carboxamide (24za). Obtained following the general procedure D from 22za. White solid (35 mg, 60%); mp = 196−197 °C, [α]20D = 117.3 (c 0.024, CHCl3). 1H NMR (500 MHz, CDCl3) δ 8.87 (d, J = 7.5 Hz, 1H), 8.27 (dd, J = 8.0, 2.0 Hz, 1H), 7.52−7.45 (m, 1H), 7.23−7.15 (m, 3H), 7.14−7.08 (m, 3H), 7.00 (d, J = 8.5 Hz, 1H), 6.73 (d, J = 7.5 Hz, 1H), 6.58 (d, J = 8.0 Hz, 1H), 4.85−4.73 (m, 2H), 4.46−4.38 (m, 1H), 4.26−4.19 (m, 1H), 4.18−4.08 (m, 2H), 3.53 (dd, J = 9.0, 4.0 Hz, 1H), 3.41 (s, 3H), 3.32 (d, J = 5.0 Hz, 1H), 3.10 (dd, J = 14.0, 5.0 Hz, 1H), 2.86 (d, J = 5.0 Hz, 1H), 2.75 (dd, J = 14.0, 9.0 Hz, 1H), 2.03−1.75 (m, 4H), 1.56−1.26 (m, 11H) ppm. 13C NMR (125 MHz, CDCl3) δ 207.37, 171.19, 170.36, 166.22, 157.30, 136.08, 133.54, 132.54, 129.45, 128.59, 127.06, 121.34, 121.08, 112.09, 71.80, 69.11, 68.10, 59.38, 54.25, 52.61, 52.57, 52.44, 37.19, 31.12, 27.32, 26.18, 25.74, 25.32, 22.24, 16.63. ESI-MS: m/z = 580 [M + H]+. (10S,13S)-13-(Methoxymethyl)-N-((S)-1-((R)-2-methyloxiran-2-yl)-1-oxo-3-phenylpropan-2-yl)-12,15-dioxo2,3,4,5,6,7,8,9,10,11,12,13,14,15-tetradecahydrobenzo[b][1]oxa[5,8]diazacycloheptadecine-10-carboxamide (24zb). Obtained following the general procedure D from 22zb. White solid (34 mg, 57%); mp = 191−192 °C, [α]20D = 21.2 (c 0.03, CHCl3). 1H NMR (500 MHz, CDCl3) δ 8.86 (d, J = 8.0 Hz, 1H), 8.17 (dd, J = 8.0, 1.5 Hz, 1H), 7.48−7.40 (m, 1H), 7.19−7.00 (m, 7H), 6.95 (d, J = 8.5 Hz, 1H), 6.84 (d, J = 8.0 Hz, 1H), 4.80−4.74 (m, 1H), 4.73−4.67 (m, 1H), 4.39−4.33 (m, 1H), 4.20−4.14 (m, 1H), 4.11−4.01 (m, 2H), 3.48 (dd, J = 9.0, 4.0 Hz, 1H), 3.34 (s, 3H), 3.25 (d, J = 5.0 Hz, 1H), 3.02 (dd, J = 14.0, 4.5 Hz, 1H), 2.80 (d, J = 5.0 Hz, 1H), 2.69 (dd, J = 14.0, 9.0 Hz, 1H), 1.93−1.83 (m, 1H), 1.82−1.70 (m, 3H), 1.52−1.19 (m, 13H) ppm. 13C NMR (125 MHz, CDCl3) δ 207.61, 171.72, 170.56, 166.53, 157.57, 136.10, 133.72, 132.37, 129.26, 128.42, 126.89, 121.12, 119.98, 111.97, 71.49, 68.91, 67.95, 59.26, 53.92, 52.73, 52.42, 52.30, 36.78, 30.82, 29.11, 26.67, 26.24, 26.09, 25.77, 22.41, 16.42. ESI-MS: m/z = 594 [M + H]+. Methyl (13S,16S)-16-(Methoxymethyl)-15,18-dioxo5,6,7,8,9,10,13,14,15,16,17,18-dodecahydro-12H-thieno[2,3i][1,11]dioxa[4,7]diazacycloheptadecine-13-carboxylate (25a). Viscous liquid (68 mg, 76%). 1H NMR (500 MHz, CDCl3) δ 8.14 (d, J = 9.0 Hz, 1H), 7.44 (d, J = 5.5 Hz, 1H), 7.24 (d, J = 7.0 Hz, 1H), 6.87 (d, J = 5.5 Hz, 1H), 4.96−4.91 (m, 1H), 4.71−4.66 (m, 1H), 4.33−4.27 (m, 1H), 4.21−4.13 (m, 2H), 3.82 (dd, J = 10.0, 3.5 Hz, 1H), 3.73−3.68 (m, 4H), 3.57−3.50 (m, 2H), 3.42−3.36 (m, 4H), 1.95−1.85 (m, 1H), 1.80−1.69 (m, 2H), 1.63−1.54 (m, 4H), 1.48−1.40 (m, 1H) ppm. ESI-MS: m/z = 429 [M + H]+.

(6S,9S)-6-(Methoxymethyl)-N-((S)-1-((R)-2-methyloxiran-2yl)-1-oxo-3-phenylpropan-2-yl)-4,7-dioxo5,6,7,8,9,10,12,13,14,15,16,17-dodecahydro-4H-pyrazolo[5,1i][1]oxa[4,7,10]triazacyclohexadecine-9-carboxamide (24v). Obtained following the general procedure D from 22v. White solid (42 mg, 74%); mp = 115−117 °C, [α]20D = 3.7 (c 0.066, CHCl3). 1H NMR (500 MHz, CDCl3) δ 7.52 (d, J = 2.0 Hz, 1H), 7.47 (d, J = 8.5 Hz, 1H), 7.32−7.27 (m, 2H), 7.25−7.20 (m, 3H), 7.14 (d, J = 8.0 Hz, 1H), 6.93 (d, J = 5.5 Hz, 1H), 6.59 (d, J = 2.0 Hz, 1H), 5.10 (ddd, J = 14.0, 12.0, 4.5 Hz, 1H), 5.03 (td, J = 8.0, 6.0 Hz, 1H), 4.59 (dd, J = 10.0, 4.5 Hz, 1H), 4.43 (ddd, J = 6.0, 3.5, 2.5 Hz, 1H), 4.14 (ddd, J = 13.5, 5.0, 3.5 Hz, 1H), 4.03 (dd, J = 10.0, 4.5 Hz, 1H), 3.82 (dd, J = 9.5, 2.5 Hz, 1H), 3.72 (dd, J = 10.0, 4.5 Hz, 1H), 3.53 (dd, J = 9.5, 4.0 Hz, 1H), 3.45−3.38 (m, 4H), 3.37−3.27 (m, 2H), 3.15 (dd, J = 14.0, 6.0 Hz, 1H), 2.95 (dd, J = 14.0, 8.0 Hz, 1H), 2.82 (d, J = 4.9 Hz, 1H), 2.02−1.90 (m, 1H), 1.61−1.50 (m, 1H), 1.50−1.41 (m, 1H), 1.39− 1.27 (m, 5H), 1.20−1.06 (m, 2H), 1.03−0.90 (m, 1H). 13C NMR (125 MHz, CDCl3) δ 207.65, 170.09, 169.17, 161.70, 138.45, 136.22, 134.65, 129.16, 128.52, 126.95, 107.63, 72.10, 70.24, 69.21, 59.31, 59.19, 54.72, 54.22, 52.26, 51.59, 49.26, 37.37, 29.80, 29.35, 26.42, 25.66, 16.37. HRMS m/z calcd for C29H39N5O7 [M + H]+ 570.2922, found 570.2929. (6S,9S)-6-(Methoxymethyl)-N-((S)-1-((R)-2-methyloxiran-2yl)-1-oxo-3-phenylpropan-2-yl)-4,7-dioxo5,6,7,8,9,10,13,14,15,16,17,18-dodecahydro-4H,12Hpyrazolo[5,1-i][1]oxa[4,7,10]triazacycloheptadecine-9-carboxamide (24w). Obtained following the general procedure D from 22w. White solid (46 mg, 78%); mp = 130−131 °C, [α]20D = 38.5 (c 0.054, CHCl3). 1H NMR (500 MHz, CDCl3) δ 7.45−7.41 (m, 1H), 7.31−7.12 (m, 7H), 7.03−6.97 (m, 1H), 6.64 (d, J = 2.0 Hz, 1H), 5.02−4.89 (m, 2H), 4.66 (dd, J = 11.5, 5.0 Hz, 1H), 4.53−4.44 (m, 1H), 4.25−4.13 (m, 1H), 3.95−3.88 (m, 1H), 3.72 (dt, J = 8.0, 3.5 Hz, 1H), 3.68−3.63 (m, 1H), 3.54 (dd, J = 10.0, 4.0 Hz, 1H), 3.38 (s, 3H), 3.35−3.29 (m, 2H), 3.27−3.21 (m, 1H), 3.09 (dd, J = 14.0, 6.5 Hz, 1H), 2.93−2.85 (m, 1H), 2.79 (dd, J = 5.0, 2.0 Hz, 1H), 1.85− 1.73 (m, 1H), 1.65−1.55 (m, 1H), 1.37−1.16 (m, 11H). 13C NMR (125 MHz, CDCl3) δ 207.65, 169.67, 169.54, 160.77, 137.52, 135.81, 133.34, 129.37, 128.51, 127.06, 107.96, 70.92, 70.75, 69.03, 59.29, 59.16, 53.69, 52.21, 51.91, 37.70, 28.74, 28.31, 26.85, 24.32, 24.18, 18.43, 16.27. ESI-MS: m/z = 584 [M + H]+. (6S,9S)-6-(Methoxymethyl)-1-methyl-N-((S)-1-((R)-2-methyloxiran-2-yl)-1-oxo-3-phenylpropan-2-yl)-4,7- dioxo4,5,6,7,8,9,10,12,13,14,15,16,17,18-tetradecahydro-1Hpyrazolo[4,3-i][1]oxa[4,7]diazacycloheptadecine-9-carboxamide (24x). Obtained following the general procedure D from 22x. White solid (40 mg, 67%); mp = 73−75 °C, [α]20D = −182.8 (c 0.034, CHCl3). 1H NMR (500 MHz, CDCl3) δ 7.70 (s, 1H), 7.46 (d, J = 8.0 Hz, 1H), 7.30−7.14 (m, 6H), 6.66 (d, J = 6.0 Hz, 1H), 5.00− 4.93 (m, 1H), 4.62−4.56 (m, 1H), 4.42 (dt, J = 7.5, 3.0 Hz, 1H), 3.97 (dd, J = 10.0, 5.0 Hz, 1H), 3.83−3.76 (m, 4H), 3.68 (dd, J = 10.0, 5.0 Hz, 1H), 3.51 (dd, J = 9.0, 3.5 Hz, 1H), 3.47−3.37 (m, 4H), 3.35− 3.24 (m, 3H), 3.10 (dd, J = 13.5, 7.5 Hz, 1H), 2.94 (dd, J = 14.0, 6.5 Hz, 1H), 2.74 (d, J = 5.0 Hz, 1H), 2.60−2.50 (m, 1H), 1.82−1.70 (m, 1H), 1.48−1.18 (m, 11H), 1.17−1.07 (m, 1H). 13C NMR (125 MHz, CDCl3) δ 207.79, 170.07, 169.90, 164.69, 147.09, 137.68, 136.09, 129.44, 128.42, 126.92, 112.66, 70.93, 70.77, 69.06, 59.20, 59.12, 54.05, 52.10, 51.55, 37.85, 36.62, 28.03, 26.84, 26.13, 24.52, 23.70, 16.15. ESI-MS: m/z = 598 [M + H]+. (13S,16S)-16-(Methoxymethyl)-N-((S)-1-((R)-2-methyloxiran-2-yl)-1-oxo-3-phenylpropan-2-yl)-15,18-dioxo5,6,7,8,9,10,13,14,15,16,17,18-dodecahydro-12H-thieno[2,3i][1,11]dioxa[4,7]diazacycloheptadecine-13-carboxamide (24y). Obtained following the general procedure D from 22y. White solid (40 mg, 66%); mp = 56−58 °C, [α]20D = −112.9 (c 0.038, CHCl3). 1H NMR (500 MHz, CDCl3) δ 8.15 (d, J = 7.5 Hz, 1H), 7.44 (d, J = 5.5 Hz, 1H), 7.20−7.08 (m, 5H), 7.05 (d, J = 8.0 Hz, 1H), 6.89−6.83 (m, 2H), 4.81−4.70 (m, 2H), 4.60−4.51 (m, 1H), 4.39−4.31 (m, 1H), 4.20−4.13 (m, 1H), 4.11−4.04 (m, 1H), 3.67 (dd, J = 10.0, 6.0 Hz, 1H), 3.57 (dd, J = 9.0, 3.0 Hz, 1H), 3.48 (dd, J = 10.0, 3.5 Hz, 1H), 3.42−3.27 (m, 6H), 3.06 (dd, J = 14.0, 5.0 Hz, 1H), 2.82 (d, J = 5.0 Hz, 1H), 2.77 (dd, J = 13.5, 8.5 Hz, 1H), 1.94− 9200



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Methyl (13S,16S)-16-(Methoxymethyl)-2-methyl-15,18dioxo-5,6,7,8,9,10,13,14,15,16, 17,18-dodecahydro-12Hthiazolo[5,4-i][1,11]dioxa[4,7]diazacycloheptadecine-13-carboxylate (25b). Viscous liquid (68 mg, 76%). 1H NMR (500 MHz, CDCl3) δ 7.95 (d, J = 9.5 Hz, 1H), 7.21 (d, J = 7.0 Hz, 1H), 4.89− 4.83 (m, 1H), 4.69−4.63 (m, 1H), 4.59 (td, J = 10.0, 2.5 Hz, 1H), 4.51−4.45 (m, 1H), 4.16−4.09 (m, 2H), 3.84 (dd, J = 10.0, 3.0 Hz, 1H), 3.72 (s, 3H), 3.68 (dd, J = 10.0, 3.0 Hz, 1H), 3.56−3.49 (m, 2H), 3.38 (s, 3H), 2.64 (s, 3H), 1.89−1.73 (m, 2H), 1.69−1.50 (m, 5H), 1.48−1.38 (m, 1H) ppm. ESI-MS: m/z = 444 [M + H]+. General Procedure for Allyl Ester Compounds 26a, 26b Synthesis. To a three-necked flask was added allyl 2-hydroxybenzoate (0.25 g, 1.4 mmol), hydroxyl compound (1.5 mmol, 1.0 equiv), triphenylphosphine (0.39 g, 1.5 mmol), and THF (3.0 mL) under N2. The mixture was cooled to 0 °C and then added dropwise diisopropylazodicarboxylate (0.34 mL, 1.7 mmol) over 5 min. The reaction mixture was stirred at room temperature overnight and subsequently triturated with a minimal amount of cold hexanes (10 mL) and EtOAc (2 mL) to remove the majority of the triphenylphosphine oxide byproduct. The hexane mixture was then purified by flash chromatography to give corresponding compound. Allyl 2-(((6S,9S)-9-(Methoxycarbonyl)-6-(methoxymethyl)-2,2-dimethyl-4,7-dioxo-3,11,14-trioxa-5,8-diazahexadecan-16-yl)oxy)benzoate (26a). Viscous liquid (0.52 g, 64%). 1H NMR (500 MHz, CDCl3) δ 7.81 (dd, J = 8.0, 1.5 Hz, 1H), 7.57−7.51 (m, 1H), 7.35 (d, J = 5.5 Hz, 1H), 7.03−6.97 (m, 2H), 6.08−5.97 (m, 1H), 5.49−5.38 (m, 2H), 5.30−5.24 (m, 1H), 4.82−4.77 (m, 2H), 4.74−4.66 (m, 1H), 4.31−4.25 (m, 1H), 4.22−4.16 (m, 2H), 3.95 (dd, J = 10.0, 3.0 Hz, 1H), 3.91−3.84 (m, 2H), 3.79 (dd, J = 9.5, 4.0 Hz, 1H), 3.76− 3.67 (m, 6H), 3.66−3.60 (m, 2H), 3.48 (dd, J = 9.5, 6.5 Hz, 1H), 3.37 (s, 3H), 1.45 (s, 9H) ppm. ESI-MS: m/z = 569 [M + H]+. Allyl 2-(((6S,9S)-9-(Methoxycarbonyl)-6-(methoxymethyl)-2,2-dimethyl-4,7-dioxo-3,11,14,17-tetraoxa-5,8-diazanonadecan-19-yl)oxy)benzoate (26b). Viscous liquid (0.55 g, 60%). 1H NMR (500 MHz, CDCl3) δ 7.80 (dd, J = 8.0, 2.0 Hz, 1H), 7.49−7.42 (m, 1H), 7.37 (d, J = 5.5 Hz, 1H), 7.02−6.96 (m, 2H), 6.07−5.97 (m, 1H), 5.49 (d, J = 2.5 Hz, 1H), 5.45−5.38 (m, 1H), 5.29−5.23 (m, 1H), 4.79 (dt, J = 5.5, 1.5 Hz, 2H), 4.69 (dt, J = 8.0, 3.5 Hz, 1H), 4.33− 4.25 (m, 1H), 4.23−4.18 (m, 2H), 3.94 (dd, J = 10.0, 3.5 Hz, 1H), 3.92−3.87 (m, 2H), 3.79 (dd, J = 9.0, 4.0 Hz, 1H), 3.76−3.68 (m, 6H), 3.66−3.57 (m, 6H), 3.48 (dd, J = 9.0, 6.5 Hz, 1H), 3.37 (s, 3H), 1.45 (s, 9H) ppm. ESI-MS: m/z = 613 [M + H]+. General Procedure for Acid Compounds 27a, 27b Synthesis. A three-necked flask was charged with allyl ester (1.2 mmol, 1.0 equiv), Pd(PPh3)4 (69 mg, 60 μmol), and 1,3-dimethylbarbituric acid (0.19 g, 1.2 mmol, 1.0 equiv) under N2. To this mixture was added DCM (8 mL) and stirred at room temperature overnight. The reaction mixture was cooled to 0 °C and quenched with saturated NaHCO3. The water layer was acidified to pH 3 with 1 N HCl (5 mL) and extracted with EtOAc (10 mL × 2). The combined organic layer was washed with brine (8 mL × 2), dried over Na2SO4, filtrated, and concentrated in vacuum to the give acid compound for the next step without further purification. 2-(((6S,9S)-9-(Methoxycarbonyl)-6-(methoxymethyl)-2,2-dimethyl-4,7-dioxo-3,11,14-trioxa-5,8-diazahexadecan-16-yl)oxy)benzoic Acid (27a). Viscous liquid (0.37 g, 58%). 1H NMR (500 MHz, CDCl3) δ 8.14 (d, J = 7.5 Hz, 1H), 7.57−7.51 (m, 1H), 7.40 (d, J = 4.0 Hz, 1H), 7.17−7.10 (m, 1H), 7.04 (d, J = 8.0 Hz, 1H), 5.55 (d, J = 6.5 Hz, 1H), 4.76−4.70 (m, 1H), 4.39−4.30 (m, 3H), 3.94 (dd, J = 9.5, 2.5 Hz, 1H), 3.91−3.86 (m, 2H), 3.81 (dd, J = 9.5, 4.0 Hz, 1H), 3.77−3.71 (m, 4H), 3.70−3.63 (m, 4H), 3.55−3.48 (m, 1H), 3.38 (s, 3H), 1.44 (s, 9H) ppm. ESI-MS: m/z = 529 [M + H]+. 2-(((6S,9S)-9-(Methoxycarbonyl)-6-(methoxymethyl)-2,2-dimethyl-4,7-dioxo-3,11,14,17-tetraoxa-5,8-diazanonadecan-19-yl)oxy)benzoic Acid (27b). Viscous liquid (0.59 g, 86%). 1H NMR (500 MHz, CDCl3) δ 8.15 (d, J = 7.5 Hz, 1H), 7.57−7.51 (m, 1H), 7.37 (d, J = 5.5 Hz, 1H), 7.17−7.11 (m, 1H), 7.05 (d, J = 8.0 Hz, 1H), 5.48 (d, J = 5.0 Hz, 1H), 4.72−4.66 (m, 1H), 4.41−4.36 (m, 2H), 4.33−4.25 (m, 1H), 3.96−3.90 (m, 3H), 3.79 (dd, J = 9.0, 4.0 Hz, 1H), 3.76−3.64 (m, 9H), 3.62−3.58 (m, 4H), 3.49 (dd, J = 8.5, 7.0

Hz, 1H), 3.38 (s, 3H), 1.45 (s, 9H) ppm. ESI-MS: m/z = 573 [M + H]+. General Procedure for Macrocyclic Esters 28a, 28b Synthesis. To a solution of acid (0.69 mmol, 1.0 equiv) in DCM (2 mL) at 0 °C, was added TFA (1 mL) dropwise. The mixture was stirred at room temperature for 2 h and then concentrated under high vacuum to give the crude product for next step without further purification. The TFA salt (0.69 mmol, 1.0 equiv), HBTU (0.31 g, 0.83 mmol), and HOBt (0.11 g, 0.83 mmol) were dissolved in THF (138 mL) and cooled to 0 °C. The DIPEA (0.27 mL, 1.7 mmol) was added dropwise to this mixture. The ice bath was removed and the reaction mixture was stirred at room temperature for 6 h. The solvent was evaporated. The residue was diluted with 10 mL of EtOAc and then washed with saturated NH4Cl solution (8 mL × 2), saturated NaHCO3 (8 mL × 2), and brine (8 mL × 2), dried over Na2SO4, filtered, and concentrated in vacuo. Chromatographic purification of the residue provided the macrocyclic ester. Methyl (9S,12S)-12-(Methoxymethyl)-11,14-dioxo2,3,5,6,9,10,11,12,13,14-decahydro-8H-benzo[o][1,4,7]trioxa[10,13]diazacyclohexadecine-9-carboxylate (28a). Viscous liquid (0.15 g, 53%). 1H NMR (500 MHz, CDCl3) δ 8.90 (d, J = 9.5 Hz, 1H), 8.25 (dd, J = 8.0, 2.0 Hz, 1H), 7.51−7.45 (m, 1H), 7.38 (d, J = 8.5 Hz, 1H), 7.14−7.08 (m, 1H), 6.97 (d, J = 8.0 Hz, 1H), 5.13−5.07 (m, 1H), 4.84 (dt, J = 8.5, 3.0 Hz, 1H), 4.42−4.35 (m, 1H), 4.31− 4.24 (m, 1H), 4.16 (dd, J = 9.0, 2.0 Hz, 1H), 4.07 (dd, J = 10.0, 2.0 Hz, 1H), 3.96−3.89 (m, 2H), 3.86−3.79 (m, 1H), 3.75−3.66 (m, 4H), 3.62 (dd, J = 10.0, 3.0 Hz, 1H), 3.59−3.51 (m, 2H), 3.50−3.45 (m, 1H), 3.39 (s, 3H) ppm. ESI-MS: m/z = 411 [M + H]+. M e t h y l ( 1 2 S , 1 5 S ) - 1 5 - ( M e t h o x y me t h y l ) - 1 4 , 1 7 - d i o x o 2,3,5,6,8,9,12,13,14,15,16,17-dodecahydro-11H-benzo[r][1,4,7,10]tetraoxa[13,16]diazacyclononadecine-12-carboxylate (28b). Viscous liquid (0.15 g, 46%). 1H NMR (500 MHz, CDCl3) δ 8.77 (d, J = 7.5 Hz, 1H), 8.23 (d, J = 8.0 Hz, 1H), 7.49−7.42 (m, 1H), 7.30 (d, J = 7.0 Hz, 1H), 7.13−7.05 (m, 1H), 6.97 (d, J = 8.5 Hz, 1H), 4.95−4.88 (m, 1H), 4.77−4.70 (m, 1H), 4.42−4.35 (m, 1H), 4.28− 4.20 (m, 1H), 4.11−4.01 (m, 2H), 3.91−3.82 (m, 3H), 3.73−3.64 (m, 8H), 3.64−3.58 (m, 2H), 3.57−3.47 (m, 2H), 3.41 (s, 3H) ppm. ESI-MS: m/z = 455 [M + H]+. General Procedure for Compounds 30a, 30b Synthesis. To a solution of the corresponding multi oxygen atoms containing macrocyclic ester (0.14 mmol, 1.0 equiv) in acetone (0.68 mL) was added aqueous LiOH 0.3 M (1.5 equiv) dropwise. The solution was stirred at room temperature for 6 h. The reaction mixture was then acidified to pH 2 with 3 N HCl and extracted with EtOAc, washed with brine, dried (Na2SO4), filtered, and concentrated. The crude macrocyclic acid (0.10 mmol, 1.0 equiv) was redissolved in 3 mL of THF and cooled to 0 °C. It was sequentially treated with HOBt (0.12 mmol, 1.2 equiv), HBTU (0.12 mmol, 1.2 equiv), Phe-epoxyketone (23, 0.12 mmol, 1.2 equiv), and DIPEA (0.15 mmol, 1.5 equiv). After stirring at room temperature for 30 min, amine HCl salt mentioned above (1.2 mmol, 1.2 equiv) and DIPEA (3.0 mmol, 3.0 equiv), the reaction mixture was stirred at room temperature for 3 h. The solvent was evaporated, and the residue was diluted with 5 mL of EtOAc and then washed with saturated NaHCO3 (5 mL × 2) and brine (5 mL × 2), dried over Na2SO4, filtered, and concentrated in vacuo. Flash chromatography on C18 (50% to 80% MeOH in H2O) yielded the final product. (9S,12S)-12-(Methoxymethyl)-N-((S)-1-((R)-2-methyloxiran-2-yl)1-oxo-3-phenylpropan-2-yl)-11,14-dioxo-2,3,5,6,9,10,11,12,13,14decahydro-8H-benzo[o][1,4,7]trioxa[10,13]diazacyclohexadecine9-carboxamide (30a). White solid (43 mg, 77%); mp = 150−151 °C, [α]20D = 9.4 (c c 0.042, CHCl3). 1H NMR (500 MHz, CDCl3) δ 8.82 (d, J = 7.0 Hz, 1H), 8.18 (dd, J = 7.5, 2.5 Hz, 1H), 7.47 (ddd, J = 8.5, 7.5, 1.5 Hz, 1H), 7.12−6.95 (m, 9H), 4.78 (ddd, J = 7.0, 3.0, 2.5 Hz, 1H), 4.70 (td, J = 8.5, 5.5 Hz, 1H), 4.64 (dt, J = 8.5, 3.5 Hz, 1H), 4.43−4.32 (m, 2H), 4.03 (dd, J = 9.5, 2.5 Hz, 1H), 3.99 (dd, J = 9.0, 3.5 Hz, 1H), 3.95−3.89 (m, 1H), 3.78−3.72 (m, 1H), 3.67−3.61 (m, 2H), 3.55 (t, J = 8.0 Hz, 1H), 3.43−3.38 (m, 6H), 3.33 (d, J = 5.0 Hz, 1H), 3.00 (dd, J = 13.5, 5.0 Hz, 1H), 2.79 (d, J = 5.0 Hz, 1H), 2.75 (dd, J = 13.5, 8.5 Hz, 1H), 1.40 (s, 3H) ppm. 13C NMR (125 MHz, CDCl3) δ 207.10, 170.91, 169.45, 166.42, 156.87, 136.27, 133.46, 9201



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quenched using 200 μL of acetonitrile containing 1 μM compound 24b as the internal standard. After quenching, the mixtures were vortexed and centrifuged. The liquid supernatant was collected carefully without any opacity and injected into a Shimadzu LCMS2020 mass spectrometer equipped with an electrospray ionization source to determine relative peak areas of each parent compound. Caco-2 Cell Permeability Assays. Compounds were dissolved in Hank’s Balanced Salt Solution (HBSS) buffered at pH 7.4 using 25 mM HEPES. All solutions were pretempered. A buffer at pH 7.4 lacking the drug was added to the receiver side of the membrane of cells, and the drug solution (pH 7.4) was added to the donor side. The membranes were incubated in a humidified atmosphere at 37 °C. At regular time intervals (every 90 min), samples were transferred to the analysis plate and matrix matched with blank donor buffer. Acetonitrile containing internal standard (imipramine) was added to all samples, and they were analyzed by LC-MS/MS (Agilent rapid resolution HPLC, AB SCIEX 4500 MS). The normalized (analyte/ internal standard) peak areas were used to calculate the apparent permeability (Papp). The membrane integrity was assessed by calculating the Papp of Lucifer yellow (acceptable values of

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