Synthesis and Structure–Activity Relationship of C-Phenyl D-Glucitol (TP0454614) Derivatives as Selective Sodium-Dependent Glucose Cotransporter 1 (SGLT1) Inhibitors

Abstract

Sodium-glucose cotransporter 1 (SGLT1) is the primary transporter for glucose absorption from the gastrointestinal tract. While C-phenyl D-glucitol derivative SGL5213 has been reported to be a potent intestinal SGLT1 inhibitor for use in the treatment of type 2 diabetes, no SGLT1 selectivity was found in vitro (IC₅₀ 29 nM for hSGLT1 and 20 nM for hSGLT2). In this study we found a new method of synthesizing key intermediates 12 by a one-pot three-component condensation reaction and discovered C-phenyl D-glucitol 41j (TP0454614), which has >40-fold SGLT1 selectivity in vitro (IC₅₀ 26 nM for hSGLT1 and 1101 nM for hSGLT2). The results of our study have provided new insights into the structure–activity relationships (SARs) of the SGLT1 selectivity of C-glucitol derivatives.

Introduction

The number of people with type 2 diabetes (T2DM) have been steadily increasing over the past few decades, and the increase has created a public health problem. A WHO global report estimates that 422 million adults have diabetes. In the past decade, the prevalence of diabetes has been increasing more rapidly in low and middle income countries than in high income countries, so the number of people with diabetes is likely to continue to increase. In 2012, 1.5 million people died from diabetes. In addition, T2DM can cause complications in many parts of the body and may increase the overall risk of premature death. Specific complications include heart failure (HF), renal failure, blindness, and nerve damage. Since overweight and obesity are related risk factors for diabetes, a well-balanced diet and exercise are the basics of diabetes prevention. In T2DM patients, blood glucose control using drugs, as well as exercise and diet, is important to prevent and slow the progression of complications.¹⁾

Sodium-dependent glucose cotransporters (SGLTs) are emerging as new therapeutic targets for T2DM. SGLT2 is expressed in the early renal proximal tubule and reabsorbs more than 90% of the filtered glucose in the kidney (160–180 g/d).^2–4) Some SGLT2 inhibitors, including dapagliflozin,⁵⁾ canagliflozin,⁶⁾ empagliflozin,⁷⁾ ipragliflozin,⁸⁾ luseogliflozin,⁹⁾ and tofogliflozin,¹⁰⁾ have been approved for the treatment of T2DM. Recent evidence is increasing that SGLT2 inhibitors can reduce the risk for heart failure in patients with T2DM.^11–13) Potential mechanism of improved cardiac function with SGLT2 is reviewed,¹⁴⁾ but direct SGLT2 inhibition effect is unlikely considered, because SGLT2 is expressed exclusively in kidney and not in heart at all.

On the other hand, SGLT1 is highly expressed not only in the kidney and small intestine but also in various other organs and cells, such as the heart, liver, lung, skeletal muscle, and brain.¹⁵⁾ Whether SGLT1 in heart exerts protective or deleterious effect has not yet been demonstrated. Thus, we have been interested in therapeutic potential of SGLT1 and its mechanism for HF. Recent studies demonstrated that SGLT1 activation improves ischemia-reperfusion-induced cardiac injury, and increased SGLT1 gene expression is observed in hypertrophic, ischemic, and diabetic cardiomyopathy in human hearts.¹⁶⁾ On the other side, chronic SGLT1 overexpression has demonstrated to cause a phenotype similar to glycogen-storage cardiomyopathy.^17,18)

The role of SGLT1 in heart is studied using SGLT1-knockdown mice or phlorizin as a tool compound. Since knockdown-mice suppress the physiological action of SGLT1 expressed in all organs including small intestine and kidney, it cannot be ruled out that the glucose homeostasis indirectly may affect cardiac function. Phrolizin, which is a non-specific SGLT inhibitor and easily metabolized by β-glucosidase in vivo, is not always best tool compound for mechanistic analysis.

Previously, we succeeded in finding a candidate the low absorption SGLT1 inhibitor SGL5213 (IC₅₀ 29 nM/20 nM for hSGLT1/hSGLT2), which suppresses glucose absorption in the small intestine, however, SGL5213 is not SGLT1 selective inhibitor in vitro.¹⁹⁾ Thus, we thought the SGLT1-specific and metabolically stable inhibitor is better to elucidate a physiological role in vivo. In our knowledge, a chemically stable C-glycoside derivative with enhanced SGLT1 selectivity has not been reported.

We began our study with the previously reported non-selective chemical class of compound 1 as the starting compound. Since there is no crystal information on SGLT1, we optimized the SGLT1 selectivity with ligand-based drug design (Fig. 1).

Fig. 1. Synthesis of a Low-Absorbable Selective SGLT1 Inhibitor

First, we attempted to achieve selectivity by modifying R¹ in the side chain of compound 2 and the substituent R² in the benzene ring of compound 3. Introducing a substituent of appropriate size in R² and introducing an alkyl group at an appropriate position in R¹ have been found to contribute to improving selectivity. Second, because selectivity was insufficient, we modified the terminal side chain R³. We found that introducing a cyclic amine in R³ moiety improves SGLT1 selectivity, and the combination of all of these changes enabled us to achieve 40-fold selectivity.

Below we describe the synthesis and structure–activity relationships (SARs) of this new series of C-phenyl D-glucitol derivatives in our attempt to improve SGLT2/SGLT1 selectivity.

Chemistry

The synthesis of key intermediate 12 is shown in Chart 1. A description of the synthesis of key intermediate 12 has already been published.¹⁹⁾ We have now established a short-step route that includes a one-pot three-component condensation reaction. The new route does not require column purification, and it facilitates large-scale synthesis, which is a great advantage when performing derivative synthesis. To synthesize 12, commercially available 3-isopropylphenol (5) was brominated with Br₂ under acidic conditions and protected by a methoxymethyl group by using an alkyl halide under basic conditions to yield 6. Lactone 7 was added to lithium reagents prepared from 6 and n-BuLi at −78°C, and then the hydroxyl group at the anomeric position was protected with a trimethylsilyl group, using chlorotrimethylsilane (TMSCl). The resulting intermediate was treated with n-BuLi, and aldehyde 8 was added to yield 9 in a one-pot reaction. The hydroxyl group of anomeric position was converted to methyl ether under acidic condition in methanol, and the hydroxyl groups of the glucose moiety were protected with acetic anhydride and pyridine to yield 10. Without purification, the methyl glucoside and the acetoxy group at the benzyl position of compound 10 were reduced using t-butyldimethylsilane and trimethylsilyl triflate to yield aryl β-C-glucoside 11 in a stereoselective manner, and the phenol of 11 was acetylated to give crude 12, which was crystalized from 2-propanol to yield intermediate 12 as a colorless powder.

Chart 1.

(a) Br₂, AcOH, 4°C, 1 h; (b) MOMCl, DIPEA, CHCl₃, room temperature (r.t.), 1 h; (c) n-BuLi, THF, −78°C, 35 min, Then 7; (d) TMSCl, −78°C, 2 h; (e) n-BuLi, THF, −78°C, 30 min, Then 8; (f) MsOH, MeOH, Reflux, 1.5 h; (g) Ac₂O, Pyridine, r.t., 18 h; (h) tBuMe₂SiH, TMSOTf, MeCN, 4°C, 2 h; (i) Ac₂O, Pyridine, r.t., 18 h.

The synthesis of C-phenyl D-glucitol derivatives 1 and 21, 28a–28f, 29g is shown in Chart 2. Compound 15 was obtained by alkylation of N-carbobenzoxy-1,2-diaminoethane hydrochloride (13) with 2-bromoacetamide (14), followed by protection of the amino group with a t-butoxycarbonyl group. The benzyloxycarbonyl group of 15 was deprotected by using palladium hydroxide under a hydrogen atmosphere to yield 16. A Mizoroki-Heck reaction of 12 and 2,2-dimethylbut-3-enoic acid (17) under microwave irradiation conditions gave the corresponding alkenyl product 18. Compound 1 was obtained by condensing 16 and 18 using water-soluble carbodiimide hydrochloride (WSCI·HCl), followed by deprotection of the t-butoxycarbonyl group and acetoxy group.

Chart 2.

(a) 14, DIPEA, EtOH, r.t., 18 h; (b) Boc₂O, DMAP, DMF, Reflux, 18 h; (c) Pd(OH)₂/C, MeOH, r.t., 18 h; (d) 17, Pd(OAc)₂, (o-tolyl)₃P, Et₃N, MeCN, Reflux, 3 h; (e) 16, WSC HCl, HOBt H₂O, Et₃N, DMF, r.t., 18 h; (f) TFA, CHCl₃, r.t., 18 h; (g) NaOMe, MeOH, r.t., 3.5 h; (h) 19, CDI, CHCl₃, r.t., 5 h; (i) NaOMe, MeOH, r.t., 1 h; (j) 13, CHCl₃, 90°C, 25 h; (k) 22a–22g, WSC HCl, HOBt H₂O, Et₃N, DMF, r.t., 18 h; (l) Dess-Martin Periodinane, CHCl₃, r.t., 3 h; (m) 25 or 26, NaBH₃CN, MeOH, r.t., 18 h or NaB(OAc)₃H, DMF, r.t., 18 h; (n) Et₃N, MeOH/H₂O, r.t., 18 h or NaOMe, MeOH, r.t., 18 h; (o) TFA, r.t., 18 h.

Compound 20 was obtained from 18 and commercially available 1,2-diamino-2-methylpropane (19) through a condensation and deprotection sequence. Compound 21 was obtained by adding 2-bromoacetamide (14) to 20 under basic conditions. Compounds 28a–28f and 29g were obtained by the following procedure. Intermediate 18 was condensed with amine 22a–22g to give compound 23a–23g. The terminal alcohols of 23a–23g were oxidized to aldehydes 24a–24g. The reductive amination of 24a–24g was achieved by using amine 25 or its hydrochloride 26 to yield 27a–27g. Finally, the acetyl group of 27a–27g was removed under Zemplén deacetylation conditions or Et₃N in MeOH/H₂O to give 28a–28g. Only 28g was treated with trifluoroacetic acid (TFA) to obtain 29g.

The synthesis of C-phenyl D-glucitol derivatives 38a–38d is shown in Chart 3. The phenolic hydroxyl group of key intermediate 11 was methylated to give 30. The synthesis of 34b–34d in which the substituent is other than a methoxy group is as follows. The acetyl groups of 12 were removed using Et₃N in MeOH/H₂O to yield 31. The resulting phenolic hydroxyl group of 31 was selectively alkylated with 32b–32d to give 33b–33d. The residual hydroxyl groups of 33b–33d were acetylated to give 34b–34d. A Mizoroki–Heck reaction of 30, 34b–34d with 2,2-dimethylbut-3-enoic acid (17) under microwave irradiation yielded 35a–35d. 36a–36d were obtained by condensation with 22a, and oxidation by Dess-Martin periodinane. The reductive amination of 36a–36d with 25 or 26 was carried out under optimum conditions to yield 37a–37d. Finally, the acetyl groups of 37a–37d were deprotected to give 38a–38d.

Chart 3.

(a) MeI, K₂CO₃, DMF, r.t., 18 h Then 50°C, 2 h; (b) Et₃N, MeOH/H₂O, r.t., 3 d; (c) 32b–32d, K₂CO₃, DMF, 50–150°C, 3–18 h; (d) Ac₂O, Pyridine, r.t., 18 h; (e) 17, Pd(OAc)₂, (o-tolyl)₃P, Et₃N, MeCN, Reflux, 3 h; (f) 22a, WSC HCl, HOBt H₂O, Et₃N, DMF, r.t., 18 h; (g) Dess-Martin Periodinane, CHCl₃, r.t., 1 h; (h) 25 or 26, NaBH₃CN, MeOH, r.t., 18 h or NaB(OAc)₃H, DMF, AcOH, r.t., 18 h; (i) Et₃N, MeOH/H₂O, r.t., 18 h or NaOMe, MeOH, r.t., 18 h.

The synthesis of C-phenyl D-glucitol derivatives 41b–41j is shown in Chart 4. First, reductive amination of key intermediate 36a with various amines or amine hydrochloride was carried out under optimum conditions to yield 40b–40j. Next, the acetyl groups of 40b–40f, 40h–40j were removed to give 41b–41f, 41h–41j. Also 41g was obtained by N-methylating of 40f and then removing the acetyl group.

Chart 4.

(a) Amine or Amine Hydrochloride, NaBH₃CN, MeOH or NaB(OAc)₃H, DMF or CHCl₃, AcOH, r.t., 18 h; (b) Et₃N, MeOH/H₂O, r.t., 18 h; (c) aq.HCHO, NaB(OAc)₃H, CHCl₃, r.t., 6 h; (d) Et₃N, MeOH/H₂O, r.t., 16 h.

Results and Discussion

In order to improve the selectivity of benchmark compound 1, we searched for SAR by modifying the side chain R¹. In order to investigate the influence of steric hindrance, 21 and 28a with dimethyl groups were synthesized. 21 was found to have improved SGLT1 inhibitory activity and to have 3 fold greater selectivity in comparison with 1, whereas 28a had decreased SGLT1 inhibitory activity. Next, derivatives 28b–29g, in which the dimethyl group site of compound 21 was converted to cyclic alkyl, oxetane, or azetidine, were synthesized, and the effects of steric hindrance and the functional group were evaluated. The results showed that the inhibitory activity of the cycloalkane substituents on SGLT1 and SGLT2 tended to be inversely proportional to the size of the cycloalkane ring, for example cyclopropane compound 28b had twice the SGLT1 inhibitory activity but half the SGLT2 inhibitory activity of cyclohexane compound 28e. 28f is an oxetane analog of 28c, which showed similar SGLT1 inhibitory activity. On the other hand, 29g, into which a nitrogen atom had been introduced, had significantly lower SGLT1 inhibitory activity. We then decided to carry out a further optimization of compound 21, because it showed the highest selectivity among the compounds in Table 1.

Table 1. Structure–Activity Relationships of C-Phenyl D-Glucitol Derivatives

^a The IC₅₀ values for the hSGLT1 and hSGLT2 activities are the mean values of data obtained in at least two experiments. ^b Not tested.

Next, we investigated the possibility of improving selectivity by altering R² of the benzene ring. First, we synthesized 38a, because our previous findings had suggested that converting R² to a methoxy group would improve selectivity.¹⁹⁾ Testing confirmed that selectivity was significantly improved as intended, however, contrary to our expectations, the selectivity of 38b, into which an ethoxy group had been introduced, was poorer than that of 38a. We also investigated whether selectivity would be affected by introducing a hydroxyl group or an amino group at the tip of the ethoxy group. The results showed that 38c, into which a hydroxyethyl group had been introduced, had lower selectivity, and that 38d, into which an aminoethyl group had been introduced, had lost its SGLT1 inhibitory activity. From a selectivity standpoint, a methyl group was found to be the optimal R² substituent on the benzene ring. Finally, we attempted to improve selectivity by converting the end of the side chain, R³.

Table 2. Structure–Activity Relationships of C-Phenyl D-Glucitol Derivatives

Compound	R2	hSGLT1a IC50 (nM)	hSGLT2a IC50 (nM)	hSGLT2/hSGLT1
21	H	91	578	6.3
38a	Me	74	1170	15.8
38b	Et	174	1898	10.9
38c	CH2CH2OH	145	848	5.8
38d	CH2CH2NMe2	>3000	NTb	NTb

^a The IC₅₀ values for the hSGLT1 and hSGLT2 activities are the mean values of data obtained in at least two experiments. ^b Not tested.

The results of converting R³ into various structures are shown in Table 3. First, we investigated whether the SARs of 38a would change when the length of its terminal side chain was changed. The inhibitory activities of compounds 41b and 41c, which had a longer side chain than 38a, were unchanged. Next, we investigated the effect on activity of converting the terminal side chain from an amide group to a dimethyl amino group. Comparisons of 41d, 41e, and 41f, which have different lengths, showed that only the shortest compound, 41d, had improved SGLT1 inhibitory activity, and the selectivity of 41d was improved 24.8-fold. In addition, 41g, which was N-methylated 41f, had slightly improved SGLT2/SGLT1 selectivity. Inspired by these results, we decided to investigate whether selectivity could be improved by cyclization of the terminal side chain. We prepared three cyclized derivatives, 41h, 41i, and 41j, and one of them, 41j, which contains a 4-dimethyl piperidine group, showed moderate SGLT1 inhibition activity (26 nM) and 42.4-fold selectivity for SGLT1 over SGLT2. These results confirmed that selectivity is increased by enhancing SGLT1 inhibitory activity alone by introducing an N,N-dimethyl amino group at an appropriate position, as in the case of 41d. Since we were able to obtain compounds that exerted strong SGLT1 selectivity, we measured parallel artificial membrane permeation assay (PAMPA)²⁰⁾ to assess the membrane permeability of several compounds. The low PAMPA values of chemical compounds in this class suggested that their membrane permeability and absorption from the intestine would be very low. Although the administration route on animals may be limited to subcutaneous and intravenous injections, 41j could be used to elucidate SGLT1 function both in vitro and in vivo an acute or a chronic HF model as a tool compound.

Table 3. Structure–Activity Relationships of C-Phenyl D-Glucitol Derivatives

^a The IC₅₀ values for the hSGLT1 and hSGLT2 activities are the mean values of data obtained in at least two experiments. ^b Not tested.

SGLT1 expression may be beneficial following acute myocardial ischemic injury since they facilitate glucose uptake, which is the only source of ATP through anaerobic glycolysis during ischemia.²¹⁾

Administration of 41j to ischemia-reperfusion-induced cardiac injury models may indicate whether SGLT1 is essential for cardio protection or is compensated for by another facilitated-diffusion type glucose transporter 1 or 4 (GLUT1 or 4).

Conclusion

In summary, a new series of C-phenyl D-glucitol derivatives was designed and synthesized, and their selectivity (SGLT2/SGLT1) and permeability were evaluated. To achieve a class of molecules with selectivity, we conducted careful SAR studies of compounds with different R¹, R² and R³ group substitutions. The results showed that 41j had over 40-fold SGLT2/SGLT1 selectivity with 26 nM of SGLT1 inhibitory activity. There have been very few reports of SGLT1-selective compounds, and 41j (TP0454614) is the first C-glycoside type SGLT1 inhibitor discovered, it may help to elucidate the physiology of SGLT1. The further study has made it possible to investigate whether SGLT1 is a beneficial as a therapeutic target in patients with heart failure.

Experimental

Glucose Uptake Inhibition Assay

Chinese hamster ovary-K1 cells stably expressing human SGLT2 (NM_003041) or human SGLT1 (NM_000343) were used for the sodium-dependent glucose transport inhibition test. Cells were incubated for 20 min in 200 µL of pretreatment buffer solution (140 mM choline chloride, 2 mM KCl, 1 mM CaCl₂, 1 mM MgCl₂, 10 mM N-(2-hydroxyethyl)piperazine-N′-2-ethanesulfonic acid (HEPES)/5 mM Tris, pH 7.4). The pretreatment buffer was removed, and 75 µL of uptake buffer containing the test compound (methyl-α-D-glucopyranoside containing [¹⁴C] methyl-α-D-glucopyranoside [1 mM for SGLT1 inhibition, 1 mM for SGLT2 inhibition], 140 mM NaCl, 2 mM KCl, 1 mM CaCl₂, 1 mM MgCl₂, 10 mM HEPES/5 mM Tris, pH 7.4) were added. The uptake reaction was allowed to proceed at 37°C for 30 min (SGLT1) or 1 h (SGLT2). The cells were then washed twice with 200 µL of washing buffer (10 mM methyl-α-D-glucopyranoside, 140 mM choline chloride, 2 mM KCl, 1 mM CaCl₂, 1 mM MgCl₂, 10 mM HEPES/5 mM Tris, pH 7.4) and dissolved in 75 µL of 0.25 M NaOH. Opti phase Super Mix (Perkin-Elmer Corporation) was added to the solution and mixed well, and its radioactivity was measured with a Micro Beta Trilux counter (Perkin-Elmer Corporation). An uptake buffer that did not contain the test compound was used as a control. Another uptake buffer containing choline chloride instead of NaCl was also prepared as a background control. The IC₅₀ values were determined by testing each test compound at six suitable concentrations and calculating the concentration at which glucose uptake was inhibited by 50% (IC₅₀ value) compared with glucose uptake in the control (100%).

Pharmacokinetic Evaluation

Permeability measurements were made by performing PAMPA Evolution™ assays as described previously.²⁰⁾ A 96-well PAMPA sandwich plate (pION Inc.) was used for the permeability measurements. The “sandwich” filters were coated with 4 µL of GIT-0 Lipid (pION Inc.), and the donor plate was filled with 200 µL of the test compound dissolved in the PRISMA HT Universal Buffer (pION Inc.). The concentrations of the test compound in the donor solution were 25 µM for compounds 41g, 41h, 12.5 µM for compounds 41d, 41e, 41j, and 10 µM for compound 38a.

Chemistry

All solvents and reagents were obtained from commercial suppliers and used without further purification, or were prepared according to published procedures. The ¹H-NMR and ¹³C-NMR spectra were recorded using a JOEL JNM-ECA 600, Varian Inova 300, or Gemini 2000 NMR spectrometer, and all chemical shifts were reported in parts per million (ppm) relative to tetramethylsilane (TMS) as an internal standard. Mass spectra were recorded using a Shimadzu LCMS 2010 EV spectrometer. High-resolution mass spectral data were acquired using a Shimadzu LCMS-IT-TOF mass spectrometer equipped with an electrospray ionization/atmospheric pressure chemical ionization (ESI/APCI) dual ion source.

1,5-Dibromo-2-(methoxymethoxy)-4-(propan-2-yl)benzene (6)

A solution of bromine (469 g, 2.94 mol) in acetic acid (320 mL) was added dropwise over 1 min at room temperature to a solution of 3-isopropylphenol (160 g, 1.18 mol) in acetic acid (1.6 L) and stirred for 1 h so that the internal temperature did not exceed 19°C. After adding toluene (1.6 L), the mixture was ice-cooled, and a 10% aqueous sodium sulfite solution (1.0 L) was added dropwise so as not to exceed an internal temperature of 20°C. The organic layer was separated, washed twice with 10% aqueous sodium sulfite solution (1.0 L) and 10% brine (1.0 L), and then dried over anhydrous magnesium sulfate. After filtering off the desiccant, the solvent was evaporated under reduced pressure to give 2,4-dibromo-5-isopropylphenol (342 g, 99%) in the form of a pale yellow oil.

N,N-diisopropylethylamine (364 mL, 2.09 mol) was added to a solution of 2,4-dibromo-5-isopropylphenol (512 g, 1.74 mol) in chloroform (1.74 L), and the mixture was ice-cooled. Chloromethyl methyl ether (159 mL, 2.09 mol) was added dropwise over a 1-hour period, and the solution was stirred at room temperature for 1 h. The reaction solution was then ice-cooled, and after adding 1 M aqueous sodium hydroxide solution (1.5 L) dropwise, the organic layer was separated, washed with 1 M aqueous sodium hydroxide solution (1.5 L) and water (1.5 L), and dried over anhydrous magnesium sulfate. After filtering off the desiccant, the solvent was distilled off under reduced pressure. The residue obtained was purified by distillation under reduced pressure (0.93 to 1.5 hPa, 122°C. to 137°C.) to obtain the title compound (548 g, 96%) in the form of a pale yellow oil.

¹H-NMR (300 MHz, CDCl₃) δ ppm 1.22 (d, J = 6.8 Hz, 6H), 3.28 (sept, J = 6.8 Hz, 1H), 3.52 (s, 3H), 5.23 (s, 2H), 7.06 (s, 1H), 7.69 (s, 1H); MS (ESI/APCI Dual) m/z: 339[M + H]⁺.

(1S)-2,3,4,6-Tetra-O-acetyl-1,5-anhydro-1-{5-[(4-bromo-2-methylphenyl)methyl]-2-hydroxy-4-(propan-2-yl)phenyl}-D-glucitol (11)

A 2.69 M n-butyllithium hexane solution (231 mL, 0.621 mol) was added dropwise over 20 min to a tetrahydrofuran solution (2.84 L) of compound 6 (200 g, 0.592 mol) at −80° to −76°C under an argon atmosphere, and stirred at the same temperature for 35 min. A solution of 2,3,4,6-tetra-O-trimethylsilyl-D-glucono-1,5-lactone (7) (290 g, 0.621 mol) in tetrahydrofuran (800 mL) was then added dropwise over 55 min and stirred at the same temperature for 50 min. Next, trimethylchlorosilane (75.7 mL, 0.621 mol) was added dropwise over 15 min, and the mixture was stirred at the same temperature for 2 h. Then, a 2.69 M n-butyllithium hexane solution (319 mL, 0.858 mol) was added dropwise over 29 min, and the mixture was stirred at the same temperature for 40 min. Finally, a solution of 4-bromo-2-methylbenzaldehyde (130 g, 0.651 mol) in tetrahydrofuran (800 mL) was added dropwise over 54 min, and the mixture was stirred at the same temperature for 30 min. Water (2.85 L) was added to the reaction solution and warmed to room temperature. Toluene (2.0 L) was added, the organic layer was separated, and the solvent was evaporated under reduced pressure to yield crude compound 9 (546 g). Compound 9 (546 g) was dissolved in methanol (3.0 L), and after adding methanesulfonic acid (3.84 mL, 0.0592 mol), the mixture was refluxed for 1.5 h. The reaction solution was then cooled to room temperature and neutralized with triethylamine (25 mL, 0.179 mol), and the reaction mixture was concentrated. The concentrate was dissolved in toluene (1.0 L) and washed with water (0.5 L, 1.0 L), and after adding a 1 M aqueous sodium hydroxide solution (0.6 L) and toluene (1.0 L) to the organic layer, the aqueous layer was separated and washed with toluene (1.0 L, 0.5 L). The aqueous layer was acidified by addition with 10% hydrochloric acid (0.7 L) and extracted with toluene (1.0 L), and the organic layer was separated. The organic layer was then washed with 10% brine (1.0 L) and water (0.5 L), and the solvent was evaporated under reduced pressure to yield a crude intermediate (314 g). The crude intermediate (314 g) was dissolved in pyridine (1.0 L), and after adding acetic anhydride (0.8 L, 8.51 mol) to the solution, the mixture was stirred at room temperature for 18 h. The reaction solution was ice-cooled, and after adding ice (1.5 L) and toluene (1.0 L), and the mixture was stirred for 3 h. The aqueous layer was separated and extracted with toluene (1.0 L). The combined organic layer was then washed twice with 2 M hydrochloric acid (1.5 L), 5% aqueous sodium hydrogen carbonate solution (1.0 L), and 10% saline (1.0 L), and the solvent was distilled off under reduced pressure to yield compound 10 (350 g). Compound 10 (350 g) was dissolved in acetonitrile (3.4 L) and H₂O (9.1 mL, 0.506 mol), and Et₃SiH (328 mL, 2.05 mol) was added. The solution was cooled in an ice-bath, and trimethylsilyl trifluoromethanesulfonate (TMSOTf) (403 mL, 2.23 mol) was added dropwise over 85 min under ice-cooling. After stirring for 2 h at the same temperature, a 3% aqueous sodium hydrogen carbonate solution (1.92 L) was added dropwise over 40 min, and after diluting the mixture with toluene (1.0 L) and stirring for 15 min, the organic layer was separated. The aqueous layer was extracted with toluene (1.5 L), and after washing the combined organic layer with saturated aqueous sodium hydrogen carbonate solution (1.5 L), the solvent was evaporated under reduced pressure to give the title compound (392 g) in the form of a colorless liquid. ¹H-NMR (300 MHz, CDCl₃) δ ppm 1.09–1.19 (m, 6H), 1.69 (s, 3H), 1.99 (s, 3H), 2.05 (s, 3H), 2.12 (s, 3H), 2.25 (s, 3H), 2.80–2.97 (m, 1H), 3.66–3.96 (m, 3H), 4.08–4.35 (m, 2H), 4.42–4.57 (m, 1H), 5.19–5.37 (m, 3H), 6.52 (s, 1H), 6.57 (d, J = 8.1 Hz, 1H), 6.87 (s, 1H), 7.12–7.20 (m, 1H), 7.30–7.33 (m, 1H).

(1S)-2,3,4,6-Tetra-O-acetyl-1-[2-(acetyloxy)-5-[(4-bromo-2-methylphenyl)methyl]-4-(propan-2-yl)phenyl]-1,5-anhydro-D-glucitol (12)

Acetic anhydride (385 mL) was added dropwise over 10 min to a solution of compound 11 (600 g, 0.592 mol) in pyridine (770 mL) under ice-cooling. The reaction solution was raised to room temperature, stirred at the same temperature for 18 h, ice-cooled again, and quenched with ice (1.0 L). The reaction solution was then diluted with toluene (1.0 L) and stirred for 1.5 h. The organic layer was separated and washed twice with 2 M hydrochloric acid (1.25 L), 5% aqueous sodium hydrogen carbonate solution (1.0 L), 10% brine (1.0 L), and water (0.5 L). The solvent was evaporated under reduced pressure, isopropyl alcohol (1.5 L) was added, and the mixture was heated to 80°C. After stirring for 1.5 h at room temperature, the precipitate was filtered off and dried. The resulting white powder was dissolved in isopropyl alcohol (800 mL) at 80°C under heat and stirred at room temperature for 1.5 h. The precipitate was filtered off and dried, and the title compound 12 (186 g, 45%) was obtained in the form of a white powder. ¹H-NMR (300 MHz, CDCl₃) δ ppm 1.13 (d, J = 6.8 Hz, 3H), 1.14 (d, J = 6.8 Hz, 3H), 1.76 (s, 3H), 1.99 (s, 3H), 2.03 (s, 3H), 2.06 (s, 3H), 2.27 (s, 3H), 2.36 (s, 3H), 2.86–3.01 (m, 1H), 3.72–3.80 (m, 1H), 3.87 (s, 2H), 4.01–4.09 (m, 1H), 4.23–4.32 (m, 1H), 4.46–4.53 (m, 1H), 5.11–5.20 (m, 1H), 5.22–5.29 (m, 2H), 6.56–6.61 (m, 1H), 6.98–7.01 (m, 2H), 7.18–7.23 (m, 1H), 7.32–7.35 (m, 1H).

tert-Butyl (2-Amino-2-oxoethyl)(2-{[(benzyloxy)carbonyl]amino}ethyl)carbamate (15)

Compound 13 (2.51 g, 10.9 mmol) was added to a solution of compound 14 (500 mg, 3.62 mmol) and N,N-diisopropylethylamine (1.84 mL) in ethanol solution (18 mL). The reaction solution was stirred at the room temperature for 18 h. The reaction solution was evaporated under reduced pressure. The resulting residue was purified by performing silica gel column chromatography (chloroform : methanol ratio = 8 : 2) to obtain the colorless amorphous (1.6 g). Di-tert-butyl dicarbonate (4.04 g) and N,N-dimethyl-4-aminopyridine (440 mg) were added to a solution of obtained amorphous (1.55 g) in N,N-dimethylformamide solution (18 mL). The reaction solution was refluxed for 18 h. The reaction solution was diluted with ethyl acetate. The organic layer was separated and washed with 10% brine, and water. The solvent was evaporated under reduced pressure. The resulting residue was purified by performing silica gel column chromatography (hexane : ethyl acetate = 8 : 2 to 5 : 5) to obtain the title compound (463 mg, 36% in 2 steps) as a colorless amorphous. ¹H-NMR (600 MHz, CDCl₃) δ ppm 1.44 (s, 9H), 3.31–3.48 (m, 4H), 3.81 (s, 2H), 5.09 (br s, 2H), 7.28–7.44 (m, 5H); MS (ESI/APCI Dual) m/z: 374[M + Na]⁺.

tert-Butyl (2-Aminoethyl)(2-amino-2-oxoethyl)carbamate (16)

7.5% Pd(OH)₂/C (139 mg) was added to a solution of compound 15 (463 mg, 1.32 mmol) in methanol solution (13 mL) under hydrogen atomsphere. The reaction solution was stirred at the room temperature for 18 h. The mixture was filtered through Celite^®. The filtrate was evaporated under reduced pressure to give the title compound (297 mg). ¹H-NMR (600 MHz, DMSO) δ ppm 1.34–1.43 (m, 9H), 2.85–2.94 (m, 2H), 3.17 (s, 1H), 3.37–3.47 (m, 2H), 3.78 (s, 2H), 7.08–7.88 (m, 5H); MS (ESI/APCI Dual) m/z: 218[M + H]⁺.

(1S)-2,3,4,6-Tetra-O-acetyl-1-[2-(acetyloxy)-5-({4-[(1E)-3-carboxy-3-methylbut-1-en-1-yl]-2-methylphenyl}methyl)-4-(propan-2-yl)phenyl]-1,5-anhydro-D-glucitol (18)

Under an argon atmosphere, a suspension of compound 12 (216 g, 0.312 mol), 2,2-dimethyl-3-butenoic acid (53.4 g, 0.467 mol), palladium (II) acetate (3.50 g, 15.6 mmol), tri-o-tolylphosphine (9.48 g, 31.2 mmol) and triethylamine (86.9 mL, 0.623 mol) in acetonitrile (623 mL) was heated to reflux for 3 h. The reaction mixture was cooled to room temperature, diluted with chloroform (300 mL) and methanol (100 mL), and filtered through Celite ^®. The filtrate was concentrated under reduced pressure, and the resulting residue was dissolved in ethyl acetate (1.32 L). It was washed with 1M hydrochloric acid (0.96 L), 10% brine (1.2 L), and dried over anhydrous magnesium sulfate. After the desiccant was filtered off, ethyl acetate (1.2 L) was further added to the filtrate, isopropylamine (28.2 mL, 0.327 mol) was added, and the mixture was stirred from room temperature to 0°C for 1 h. The deposited precipitate was filtered to obtain an isopropylamine salt of intermediate. This salt was dissolved in ethyl acetate (1.2 L) and 1 M hydrochloric acid (500 mL) and stirred for 30 min, and the organic layer was separated. The organic layer was washed with 10% brine (500 mL) and dried over anhydrous magnesium sulfate. After the desiccant was filtered off, the solvent was distilled off under reduced pressure to obtain the title compound (207 g, 88%) as a colorless amorphous. ¹H-NMR (300 MHz, CDCl₃) δ ppm 1.13 (d, J = 6.8 Hz, 3H), 1.14 (d, J = 6.8 Hz, 3H), 1.43 (s, 6H), 1.76 (s, 3H), 1.99 (s, 3H), 2.03 (s, 3H), 2.05 (s, 3H), 2.28 (s, 3H), 2.37 (s, 3H), 2.98 (spt, J = 6.8 Hz, 1H), 3.70–3.80 (m, 1H), 3.91 (s, 2H), 4.05 (dd, J = 12.4, 2.2 Hz, 1H), 4.28 (dd, J = 12.4, 4.4 Hz, 1H), 4.43–4.50 (m, 1H), 5.11–5.20 (m, 1H), 5.22–5.33 (m, 2H), 6.33–6.49 (m, 2H), 6.68 (d, J = 7.9 Hz, 1H), 6.96 (s, 1H), 6.99 (s, 1H), 7.06–7.14 (m, 1H), 7.23 (d, J = 1.4 Hz, 1H); MS (ESI/APCI Dual) m/z: 747[M + Na]⁺.

(1S)-1-[5-({4-[(1E)-4-({2-[(2-Amino-2-oxoethyl)amino]ethyl}amino)-3,3-dimethyl-4-oxobut-1-en-1-yl]-2-methylphenyl}methyl)-2-hydroxy-4-(propan-2-yl)phenyl]-1,5-anhydro-D-glucitol (1)

Compound 16 (285 mg, 1.52 mmol), 1-hydroxy-1H-benzotriazole hydrate (205 mg), triethylamine (420 µL), water soluble carbodiimide hydrochloride (290 mg) were added to a solution of compound 18 (731 mg, 1.01 mmol) in N,N-dimethylformamide solution (5 mL). The reaction solution was stirred at the room temperature for 18 h. The reaction solution was diluted with ethyl acetate. The organic layer was separated and washed with 10% brine, and water. The solvent was evaporated under reduced pressure. The resulting residue was purified by performing silica gel column chromatography (hexane : ethyl acetate = 2 : 1 to 1 : 2) to obtain the crude compound (830 mg) as a colorless amorphous. Trifluoroacetic acid (1.2 mL) was added to a solution of obtained crude compound (830 mg) in chloroform solution (9 mL). The reaction solution was stirred at the room temperature for 18 h. The solvent was evaporated under reduced pressure to give the crude compound (1.5 g). 4.88 M sodium methoxide (1.1 mL) was add to a solution of the crude compound (1.5 g) in methanol solution (9 mL). The reaction solution was stirred at the room temperature for 3.5 h. The solvent was evaporated under reduced pressure. The resulting residue was purified by performing NH silica gel column chromatography (ethanol : H₂O = 9 : 1 to 2 : 1) to obtain the title compound (652 mg, 76% in 3 steps) as a colorless amorphous. ¹H-NMR (600 MHz, CD₃OD) δ ppm 1.08–1.12 (m, 6H), 1.37 (s, 6H), 2.30 (s, 3H), 2.66–2.70 (m, 2H), 2.90–2.96 (m, 1H), 3.21–3.23 (m, 2H), 3.27–3.30 (m, 2H), 3.37–3.40 (m, 2H), 3.44–3.48 (m, 1H), 3.52–3.56 (m, 1H), 3.66–3.70 (m, 1H), 3.82–3.86 (m, 1H), 3.88 (s, 2H), 4.44–4.48 (m, 1H), 6.33–6.39 (m, 1H), 6.44–6.50 (m, 1H), 6.73–6.78 (m, 1H), 6.80 (s, 1H), 6.95 (s, 1H), 7.08–7.12 (m, 1H), 7.23 (s, 1H); ¹³C-NMR (126 MHz, CD₃OD) δ ppm 19.97 (s) 24.09 (s) 24.20 (s) 25.95 (s) 30.30 (s) 36.31 (s) 40.59 (s) 45.92 (s) 49.84 (s) 52.34 (s) 63.09 (s) 71.89 (s) 75.51 (s) 79.17 (s) 80.17 (s) 82.41 (s) 114.28 (s) 123.95 (s) 125.04 (s) 129.14 (s) 129.24 (s) 130.23 (s) 130.26 (s) 131.77 (s) 134.72 (s) 136.33 (s) 137.54 (s) 140.70 (s) 149.64 (s) 155.64 (s) 177.00 (s) 179.71 (s); HR-MS ESI/APCI dual m/z: 614.3454 [M + H]⁺ (calcd for C₃₃H₄₇N₃O₈: 614.3436).

(1S)-1-{5-[(4-{(1E)-4-[(2-Amino-2-methylpropyl)amino]-3,3-dimethyl-4-oxobut-1-en-1-yl}-2-methylphenyl)methyl]-2-hydroxy-4-(propan-2-yl)phenyl}-1,5-anhydro-D-glucitol (20)

Compound 19 (128 mg, 1.45 mmol) was added to a solution of componud 18 (1.0 g, 1.38 mmol) and 1,1′-carbonylbis-1H-imidazole (235 mg) in chloroform solution (5 mL). The reaction solution was stirred at the room temperature for 5 h. The reaction mixture was twice with 10% K₂CO₃, and the organic layer was dried over MgSO₄. The solvent was evaporated under reduced pressure. The resulting residue was purified by performing NH silica gel column chromatography (chloroform) to obtain the crude compound (0.7 g). To a solution of the crude compound (0.7 g) in methanol solution (5 mL), 4.88M sodium methoxide (1.1 mL) was added. The reaction solution was stirred at the room temperature for 1 h. After acetic acid (605 µL) was added, the solvent was evaporated under reduced pressure. The resulting residue was purified by performing NH silica gel column chromatography (ethyl acetate : ethanol : H₂O = 30 : 2 : 1 to 10 : 2 : 1) to obtain the title compound (399 mg, 50% in 2 steps). ¹H-NMR (600 MHz, CD₃OD) δ ppm 1.05 (s, 6H), 1.09 (d, J = 6.88 Hz, 6H), 1.39 (s, 6H), 2.31 (s, 3H), 2.87–2.94 (m, 1H), 3.13 (s, 2H), 3.35–3.40 (m, 2H), 3.43–3.49 (m, 1H), 3.51–3.56 (m, 1H), 3.65–3.71 (m, 1H), 3.82–3.89 (m, 3H), 4.47 (d, J = 9.63 Hz, 1H), 6.38 (d, J = 16.05 Hz, 1H), 6.46–6.52 (m, 1H), 6.75 (d, J = 7.79 Hz, 1H), 6.80 (s, 1H), 6.96 (s, 1H), 7.10 (d, J = 7.79 Hz, 1H), 7.23 (s, 1H); MS (ESI/APCI Dual) m/z: 585[M + H]⁺, 583[M-H]⁻.

(1S)-1-[5-({4-[(1E)-4-({1-[(2-Amino-2-oxoethyl)amino]-2-methylpropan-2-yl}amino)-3,3-dimethyl-4-oxobut-1-en-1-yl]-2-methylphenyl}methyl)-2-hydroxy-4-(propan-2-yl)phenyl]-1,5-anhydro-D-glucitol (21)

Compound 20 (0.20 g, 0.34 mmol) was addded to a solution of compound 13 (213 mg, 1.55 mmol) in N,N-dimethylformamide solution (1 mL). The reaction solution was stirred at 90°C for 25 h. The solvent was evaporated under reduced pressure. The resulting residue was purified by performing HPLC column chromatography (Waters sunfine C-18, MeOH : 0.1% Et₃N = 3 : 7, 20 mL/min) to obtain the title compound (49 mg, 22%). ¹H-NMR (600 MHz, CD₃OD) δ ppm 1.07–1.14 (m, 6H), 1.33 (s, 6H), 1.36 (s, 6H), 2.31 (s, 3H), 2.77–2.84 (m, 2H), 2.90–2.96 (m, 1H), 3.31–3.35 (m, 2H), 3.37–3.40 (m, 2H), 3.44–3.48 (m, 1H), 3.52–3.57 (m, 1H), 3.66–3.70 (m, 1H), 3.82–3.87 (m, 1H), 3.89 (s, 2H), 4.44–4.48 (m, 1H), 6.37 (d, J = 16.0 Hz, 1H), 6.50 (d, J = 16.0 Hz, 1H), 6.74–6.78 (m, 1H), 6.80 (s, 1H), 6.96 (s, 1H), 7.11 (d, J = 7.8 Hz, 1H), 7.23 (s, 1H); ¹³C-NMR (126 MHz, CD₃OD) δ ppm 19.97 (s) 24.09 (s) 24.21 (s) 25.35 (s) 25.91 (s) 30.32 (s) 36.31 (s) 46.51 (s) 52.35 (s) 54.50 (s) 59.25 (s) 63.09 (s) 71.91 (s) 75.48 (s) 79.21 (s) 80.17 (s) 82.42 (s) 114.29 (s) 117.25 (s) 119.58 (s) 123.95 (s) 125.02 (s) 129.14 (s) 129.21 (s) 130.26 (s) 130.64 (s) 131.82 (s) 134.78 (s) 136.24 (s) 137.63 (s) 140.84 (s) 149.67 (s) 155.66 (s) 163.09 (s) 163.36 (s); HR-MS ESI/APCI Dual m/z: 642.3730 [M + H]⁺ (calcd for C₃₅H₅₁N₃O₈: 642.3749).

(1S)-2,3,4,6-Tetra-O-acetyl-1-[2-(acetyloxy)-5-[(4-{(1E)-3,3-dimethyl-4-[(2-methyl-1-oxopropan-2-yl)amino]-4-oxobut-1-en-1-yl}-2-methylphenyl)methyl]-4-(propan-2-yl)phenyl]-1,5-anhydro-D-glucitol (24a)

Compound 22a (184 mg, 2.07 mmol), 1-hydroxy-1H-benzotriazole hydrate (242 mg), water soluble carbodiimide hydrochloride (342 mg) were added to a solution of compound 18 (1.0 g, 1.38 mmol) in N,N-dimethylformamide solution (30 mL). The reaction solution was stirred at the room temperature for 18 h. Water was added and the mixture was extracted with ethyl acetate. The organic layer was washed with saturated aqueous sodium hydrogen carbonate solution and saturated brine, and dried over anhydrous magnesium sulfate. After the desiccant was filtered off, the solvent was distilled off under reduced pressure to obtain the crude intermediate (1.05 g). A chloroform suspension (20 mL) of obtained intermediate (660 mg, 0.830 mmol) and Dess–Martin periodinane (422 mg, 1.25 mmol) was stirred at room temperature for 3 h. After the solvent was distilled off under reduced pressure, the residue was purified by silica gel column chromatography (hexane : ethyl acetate = 4 : 1 to 1 : 4) to obtain the title compound (602 mg, 85% over 2 steps) as a yellow amorphous. ¹H-NMR (600 MHz, CDCl₃) δ ppm 1.09–1.18 (m, 6H), 1.33 (s, 6H), 1.39 (s, 6H), 1.78 (s, 3H), 1.99 (s, 3H), 2.03 (s, 3H), 2.04 (s, 3H), 2.31 (s, 3H), 2.37 (s, 3H), 2.98 (spt, J = 6.9 Hz, 1H), 3.73–3.80 (m, 1H), 3.89–3.98 (m, 2H), 4.06 (dd, J = 12.4, 2.3 Hz, 1H), 4.26 (dd, J = 12.4, 4.6 Hz, 1H), 4.46–4.53 (m, 1H), 5.11–5.18 (m, 1H), 5.23–5.30 (m, 2H), 6.13 (br s, 1H), 6.33 (d, J = 16.5 Hz, 1H), 6.53 (d, J = 16.5 Hz, 1H), 6.71 (d, J = 7.8 Hz, 1H), 6.99–7.01 (m, 2H), 7.13 (d, J = 7.8 Hz, 1H), 7.26 (s, 1H), 9.34 (s, 1H).

(1S)-2,3,4,6-Tetra-O-acetyl-1-[2-(acetyloxy)-5-[(4-{(1E)-4-[(1-formylcyclopropyl)amino]-3,3-dimethyl-4-oxobut-1-en-1-yl}-2-methylphenyl)methyl]-4-(propan-2-yl)phenyl]-1,5-anhydro-D-glucitol (24b)

Compound 18 (500 mg, 0.69 mmol) and compound 22b (72 mg, 0.83 mmol) were used as starting materials and synthesized in the same manner as compound 24a to give the title compound (305 mg, 98% for 2 steps) as a colorless amorphous. ¹H-NMR (300 MHz, CDCl₃) δ ppm 1.10–1.16 (m, 6H), 1.42 (s, 6H), 1.46–1.55 (m, 4H), 1.78 (s, 3H), 1.99 (s, 3H), 2.03 (s, 3H), 2.05 (s, 3H), 2.31 (s, 3H), 2.37 (s, 3H), 2.91–3.06 (m, 1H), 3.72–3.80 (m, 1H), 3.91–3.95 (m, 2H), 4.01–4.08 (m, 1H), 4.20–4.28 (m, 1H), 4.46–4.52 (m, 1H), 5.09–5.18 (m, 1H), 5.22–5.29 (m, 2H), 6.26–6.30 (m, 1H), 6.35 (d, J = 16.2 Hz, 1H), 6.55 (d, J = 16.2 Hz, 1H), 6.68–6.74 (m, 1H), 7.00 (s, 2H), 7.10–7.16 (m, 1H), 7.26(s, 1H), 9.06 (s, 1H); MS (ESI/APCI Dual) m/z: 792[M + H]⁺, 814[M + Na]⁺.

(1S)-2,3,4,6-Tetra-O-acetyl-1-[2-(acetyloxy)-5-[(4-{(1E)-4-[(1-formylcyclobutyl)amino]-3,3-dimethyl-4-oxobut-1-en-1-yl}-2-methylphenyl)methyl]-4-(propan-2-yl)phenyl]-1,5-anhydro-D-glucitol (24c)

Compound 18 (1.0 g, 1.38 mmol) and compound 22c (210 mg, 2.07 mmol) were used as starting materials and synthesized in the same manner as compound 24a to give the title compound (355 mg, 32% for 2 steps) as a colorless amorphous. ¹H-NMR (300 MHz, CDCl₃) δ ppm 1.10–1.19 (m, 6H), 1.40 (s, 6H), 1.78 (s, 3H), 1.91–2.08 (m, 11H), 2.22–2.40 (m, 8H), 2.42–2.56 (m, 2H), 2.91–3.05 (m, 1H), 3.70–3.81 (m, 1H), 3.94 (s, 2H), 4.05 (dd, J = 12.4, 1.9 Hz, 1H), 4.26 (dd, J = 12.4, 4.5 Hz, 1H), 4.44–4.55 (m, 1H), 5.10–5.20 (m, 1H), 5.21–5.33 (m, 2H), 6.28–6.41 (m, 2H), 6.49–6.61 (m, 1H), 6.71 (d, J = 7.9 Hz, 1H), 7.00 (s, 2H), 7.14 (d, J = 7.9 Hz, 1H), 7.27 (s, 1H), 9.55 (s, 1H); MS (ESI/APCI Dual) m/z: 806 [M + H]⁺, 828[M + Na]⁺.

(1S)-2,3,4,6-Tetra-O-acetyl-1-[2-(acetyloxy)-5-[(4-{(1E)-4-[(1-formylcyclopentyl)amino]-3,3-dimethyl-4-oxobut-1-en-1-yl}-2-methylphenyl)methyl]-4-(propan-2-yl)phenyl]-1,5-anhydro-D-glucitol (24d)

Compound 18 (1.0 g, 1.38 mmol) and compound 22d (207 mg, 1.79 mmol) were used as starting materials and synthesized in the same manner as compound 24a to give the title compound (850 mg, 71% for 2 steps) as a colorless amorphous. ¹H-NMR (300 MHz, CDCl₃) δ ppm 1.10–1.18 (m, 6H), 1.39 (s, 6H), 1.64–2.18 (m, 8H), 1.77 (s, 3H), 1.99 (s, 3H), 2.03 (s, 3H), 2.05 (s, 3H), 2.32 (s, 3H), 2.37 (s, 3H), 2.89–3.04 (m, 1H), 3.71–3.81 (m, 1H), 3.93 (s, 2H), 4.01–4.10 (m, 1H), 4.25–4.30 (m, 1H), 4.45–4.53 (m, 1H), 5.10–5.20 (m, 1H), 5.23–5.30 (m, 2H), 6.18 (s, 1H), 6.26–6.39 (m, 1H), 6.47–6.60 (m, 1H), 6.70 (d, J = 7.6 Hz, 1H), 6.96–7.05 (m, 2H), 7.10–7.16 (m, 1H), 7.23–7.26 (m, 1H), 9.43 (s, 1H); MS (ESI/APCI Dual) m/z: 820[M + H]⁺, 842[M + Na]⁺.

(1S)-2,3,4,6-Tetra-O-acetyl-1-[2-(acetyloxy)-5-[(4-{(1E)-4-[(1-formylcyclohexyl)amino]-3,3-dimethyl-4-oxobut-1-en-1-yl}-2-methylphenyl)methyl]-4-(propan-2-yl)phenyl]-1,5-anhydro-D-glucitol (24e)

Compound 18 (1.0 g, 1.38 mmol) and compound 22e (267 mg, 2.07 mmol) were used as starting materials and synthesized in the same manner as compound 24a to give the title compound (265 mg, 23% for 2 steps) as a light brown amorphous.MS (ESI/APCI Dual) m/z: 834[M + H]⁺, 856[M + Na]⁺.

(1S)-2,3,4,6-Tetra-O-acetyl-1-[2-(acetyloxy)-5-[(4-{(1E)-4-[(3-formyloxetan-3-yl)amino]-3,3-dimethyl-4-oxobut-1-en-1-yl}-2-methylphenyl)methyl]-4-(propan-2-yl)phenyl]-1,5-anhydro-D-glucitol (24f)

Compound 18 (815 mg, 1.12 mmol) and compound 22d (174 mg, 1.69 mmol) were used as starting materials and synthesized in the same manner as compound 24a to give the title compound (483 mg, 69% for 2 steps) as a colorless amorphous. ¹H-NMR (300 MHz, CDCl₃) δ ppm 1.06–1.10 (m, 6H), 1.43 (s, 6H), 1.79 (s, 3H), 1.98 (s, 3H), 2.03 (s, 3H), 2.05 (s, 3H), 2.31 (s, 3H), 2.36 (s, 3H), 2.93–3.06 (m, 1H), 3.71–3.80 (m, 1H), 3.93 (s, 2H), 4.04 (dd, J = 12.3, 2.2 Hz, 1H), 4.23 (dd, J = 12.4, 4.6 Hz, 1H), 4.48–4.52 (m, 1H), 4.76 (d, J = 6.7 Hz, 2H), 4.87 (d, J = 6.7 Hz, 2H), 5.08–5.18 (m, 1H), 5.18–5.33 (m, 2H), 6.36 (d, J = 16.0 Hz, 1H), 6.59 (d, J = 16.0 Hz, 1H), 6.60 (br s, 1H), 6.73 (d, J = 7.9 Hz, 1H), 6.98 (s, 1H), 7.00 (s, 1H), 7.15 (d, J = 7.9 Hz, 1H), 7.27 (s, 1H), 9.74 (s, 1H); MS (ESI/APCI Dual) m/z: 808[M + H]⁺.

(1S)-2,3,4,6-Tetra-O-acetyl-1-[2-(acetyloxy)-5-({4-[(1E)-4-{[1-(tert-butoxycarbonyl)-3-formylazetidin-3-yl]amino}-3,3-dimethyl-4-oxobut-1-en-1-yl]-2-methylphenyl}methyl)-4-(propan-2-yl)phenyl]-1,5-anhydro-D-glucitol (24g)

Compound 18 (629 mg, 0.867 mmol) and compound 22g (228 mg, 1.13 mmol) were used as starting materials and synthesized in the same manner as compound 24a to give the title compound (556 mg, 71% for 2 steps) as a colorless amorphous. ¹H-NMR (300 MHz, CDCl₃) δ ppm 1.15 (dd, J = 6.84, 4.20 Hz, 6H), 1.44 (s, 6H), 1.78 (s, 3H), 1.99 (s, 3H), 2.02–2.05 (m, 6H), 2.31 (s, 3H), 2.37 (s, 3H), 2.88 (s, 1H), 2.93–3.04 (m, 2H), 3.72–3.81 (m, 1H), 3.94 (s, 2H), 4.04 (d, J = 9.17 Hz, 2H), 4.18–4.29 (m, 3H), 4.47–4.52 (m, 1H), 5.10–5.30 (m, 3H), 6.30–6.39 (m, 1H), 6.53–6.62 (m, 2H), 6.72 (d, J = 7.93 Hz, 1H), 6.99 (d, J = 1.71 Hz, 2H), 7.14 (dd, J = 8.16, 1.63 Hz, 1H), 9.58 (s, 1H); MS (ESI/APCI Dual) m/z: 807[M + H-Boc]⁺.

(1S)-1-[5-({4-[(1E)-4-({2-[(2-Amino-2-oxoethyl)amino]-2-methylpropyl}amino)-3,3-dimethyl-4-oxobut-1-en-1-yl]-2-methylphenyl}methyl)-2-hydroxy-4-(propan-2-yl)phenyl]-1,5-anhydro-D-glucitol (28a)

Sodium cyanoborohydride (32 mg) was added to a solution of compound 24a (300 mg, 0.392 mmol) and compound 25 (44 mg, 0.588 mmol) in methanol solution (4 mL). The reaction solution was stirred at the room temperature for 18 h. After the solvent was distilled off under reduced pressure, water was added and the mixture was extracted with ethyl acetate. The organic layer was washed with saturated aqueous sodium hydrogen carbonate solution and saturated brine, and dried over anhydrous magnesium sulfate. After the desiccant was filtered off, the solvent was distilled off under reduced pressure. The residue was purified by silica gel column chromatography (chloroform : methanol = 99 : 1 to 9 : 1) to obtain the crude intermediate (43 mg). The crude intermediate (43 mg) and 4.88M sodium methoxide (64 µL) in methanol solution (0.5 mL) was stirred at the room temperature for 18 h. After the solvent was distilled off under reduced pressure, the residue was purified by NH silica gel column chromatography (ethyl acetate : ethanol : water = 15 : 2 : 1 to 7 : 2 : 1) to obtain the title compound (8 mg, 3% in 2steps) as a colorless amorphous. ¹H-NMR (500 MHz, CD₃OD) δ ppm 1.01 (s, 6H), 1.09 (s, 3H), 1.10 (s, 3H), 1.39 (s, 6H), 2.30 (s, 3H), 2.88–2.98 (m, 1H), 3.13 (s, 2H), 3.20 (s, 2H), 3.36–3.42 (m, 2H), 3.43–3.49 (m, 1H), 3.51–3.57 (m, 1H), 3.65–3.71 (m, 1H), 3.81–3.86 (m, 1H), 3.88 (s, 2H), 4.46 (d, J = 9.56 Hz, 1H), 6.36–6.42 (m, 1H), 6.48–6.55 (m, 1H), 6.75 (d, J = 8.03 Hz, 1H), 6.80 (s, 1H), 6.96 (s, 1H), 7.12 (dd, J = 8.03, 1.53 Hz, 1H), 7.23 (s, 1H); ¹³C-NMR (126 MHz, CD₃OD) δ ppm 19.98 (s) 24.10 (s) 24.21 (s) 25.14 (s) 25.96 (s) 30.31 (s) 36.32 (s) 45.81 (s) 46.09 (s) 48.42 (s) 54.99 (s) 63.12 (s) 71.91 (s) 75.53 (s) 79.14 (s) 80.18 (s) 82.43 (s) 114.28 (s) 123.98 (s) 124.99 (s) 129.16 (s) 129.19 (s) 130.24 (s) 130.49 (s) 131.78 (s) 134.84 (s) 136.28 (s) 137.58 (s) 140.79 (s) 149.62 (s) 155.68 (s) 178.02 (s) 179.46 (s); HR-MS ESI/APCI Dual m/z: 642.3759 [M + H]⁺ (calcd for C₃₅H₅₁N₃O₈: 642.3749).

(1S)-1-{5-[(4-{(1E)-4-[(1-{[(2-Amino-2-oxoethyl)amino]methyl}cyclopropyl)amino]-3,3-dimethyl-4-oxobut-1-en-1-yl}-2-methylphenyl)methyl]-2-hydroxy-4-(propan-2-yl)phenyl}-1,5-anhydro-D-glucitol (28b)

Sodium triacetoxyborohydride (30 mg) was added to a solution of compound 24b (100 mg, 0.126 mmol) and compound 26 (21 mg, 0.192 mmol) in N,N-dimethylformamide solution (4 mL). The reaction solution was stirred at the room temperature for 18 h. Water was added and the mixture was extracted with chloroform. The organic layer was washed with saturated aqueous sodium hydrogen carbonate solution and dried over anhydrous magnesium sulfate. After the desiccant was filtered off, the solvent was distilled off under reduced pressure. The residue was purified by silica gel column chromatography (chloroform : methanol = 99 : 1 to 50 : 50) to obtain the crude intermediate (62 mg). The crude intermediate (62 mg) in methanol : triethylamine : water (5 : 1 : 1) solution (2 mL) was stirred at the room temperature for 18 h. After the solvent was distilled off under reduced pressure, the residue was purified by OH silica gel column chromatography (chloroform : methanol = 100 : 0 to 50 : 50) to obtain the title compound (30 mg, 28% in 2steps) as a pale brown amorphous. ¹H-NMR (600 MHz, CD₃OD) δ ppm 0.71–0.80 (m, 4H), 1.10 (d, J = 6.9 Hz, 6H), 1.34 (s, 6H), 2.30 (s, 3H), 2.72–2.77 (m, 2H), 2.89–2.96 (m, 1H), 3.30 (s, 1H), 3.35–3.40 (m, 2H), 3.43–3.48 (m, 1H), 3.51–3.56 (m, 1H), 3.65–3.70 (m, 1H), 3.82–3.86 (m, 1H), 3.86–3.91 (s, 2H), 4.46 (d, J = 9.2 Hz, 1H), 6.35 (d, J = 16.0 Hz, 1H), 6.45 (d, J = 16.0 Hz, 1H), 6.72–6.78 (m, 1H), 6.78–6.82 (s, 1H), 6.91–6.97 (s, 1H), 7.07–7.14 (m, 1H), 7.20–7.26 (s, 1H); ¹³C-NMR (126 MHz, CD₃OD) δ ppm 13.36 (s) 19.97 (s) 24.09 (s) 24.21 (s) 25.85 (s) 30.30 (s) 33.61 (s) 36.31 (s) 45.84 (s) 51.94 (s) 56.33 (s) 63.11 (s) 71.90 (s) 75.53 (s) 79.14 (s) 80.18 (s) 82.42 (s) 114.26 (s) 123.98 (s) 125.04 (s) 129.14 (s) 129.21 (s) 130.16 (s) 130.23 (s) 131.75 (s) 134.69 (s) 136.32 (s) 137.55 (s) 140.72 (s) 149.61 (s) 155.65 (s) 175.78 (s) 180.63 (s); HR-MS ESI/APCI Dual m/z: 640.3598 [M + H]⁺ (calcd for C₃₅H₄₉N₃O₈: 640.3592).

(1S)-1-{5-[(4-{(1E)-4-[(1-{[(2-Amino-2-oxoethyl)amino]methyl}cyclobutyl)amino]-3,3-dimethyl-4-oxobut-1-en-1-yl}-2-methylphenyl)methyl]-2-hydroxy-4-(propan-2-yl)phenyl}-1,5-anhydro-D-glucitol (28c)

Compound 24c (135 mg, 0.167 mmol) and compound 26 (25 mg, 0.226 mmol) were used as starting materials and synthesized in the same manner as compound 28b to give the title compound (80 mg, 55% for 2 steps) as a colorless amorphous. ¹H-NMR (600 MHz, CD₃OD) δ ppm 1.10 (d, J = 6.9 Hz, 6H), 1.36 (s, 6H), 1.75–1.91 (m, 2H), 2.07–2.12 (m, 2H), 2.21–2.28 (m, 2H), 2.30 (s, 3H), 2.89–2.94 (m, 3H), 3.24 (s, 2H), 3.29–3.31 (m, 1H), 3.36–3.42 (m, 2H), 3.46 (t, J = 8.7 Hz, 1H), 3.55 (t, J = 8.7 Hz, 1H), 3.68 (dd, J = 11.9, 5.0 Hz, 1H), 3.84 (dd, J = 11.9, 1.8 Hz, 1H), 3.88 (s, 2H), 4.46 (d, J = 9.6 Hz, 1H), 6.36–6.41 (m, 1H), 6.46–6.51 (m, 1H), 6.75 (d, J = 7.8 Hz, 1H), 6.80 (s, 1H), 6.96 (s, 1H), 7.11 (dd, J = 7.8, 0.9 Hz, 1H), 7.24 (d, J = 0.9 Hz, 1H); ¹³C-NMR (126 MHz, CD₃OD) δ ppm 15.85 (s) 19.98 (s) 24.10 (s) 24.21 (s) 25.89 (s) 30.31 (s) 32.22 (s) 36.31 (s) 46.06 (s) 53.06 (s) 55.86 (s) 58.39 (s) 60.30 (s) 63.11 (s) 71.90 (s) 75.52 (s) 79.15 (s) 80.18 (s) 82.43 (s) 114.27 (s) 123.97 (s) 125.03 (s) 129.13 (s) 129.20 (s) 130.24 (s) 130.32 (s) 131.78 (s) 134.91 (s) 136.32 (s) 137.57 (s) 140.74 (s) 149.62 (s) 155.66 (s) 176.88 (s) 178.56 (s); HR-MS ESI/APCI Dual m/z: 654.3757 [M + H]⁺ (calcd for C₃₆H₅₁N₃O₈: 654.3749).

(1S)-1-{5-[(4-{(1E)-4-[(1-{[(2-Amino-2-oxoethyl)amino]methyl}cyclopentyl)amino]-3,3-dimethyl-4-oxobut-1-en-1-yl}-2-methylphenyl)methyl]-2-hydroxy-4-(propan-2-yl)phenyl}-1,5-anhydro-D-glucitol (28d)

Compound 24d (220 mg, 0.268 mmol) and compound 25 (30 mg, 0.402 mmol) were used as starting materials and synthesized in the same manner as compound 28a to give the title compound (16 mg, 9% for 2 steps) as a colorless amorphous. ¹H-NMR (600 MHz, CD₃OD) δ ppm 1.07–1.13 (m, 6H), 1.32–1.38 (m, 6H), 1.57–1.76 (m, 6H), 1.93–2.02 (m, 2H), 2.31 (s, 3H), 2.82 (s, 2H), 2.92 (s, 1H), 3.19 (s, 2H), 3.35–3.41 (m, 2H), 3.43–3.49 (m, 1H), 3.52–3.57 (m, 1H), 3.65–3.71 (m, 1H), 3.81–3.87 (m, 1H), 3.88 (s, 2H), 4.44–4.49 (m, 1H), 6.36–6.42 (m, 1H), 6.48–6.53 (m, 1H), 6.75 (d, J = 7.8 Hz, 1H), 6.80 (s, 1H), 6.96 (s, 1H), 7.08–7.13 (m, 1H), 7.23 (s, 1H); ¹³C-NMR (126 MHz, CD₃OD) δ ppm 19.98 (s) 24.10 (s) 24.21 (s) 25.07 (s) 25.90 (s) 30.32 (s) 36.33 (s) 37.14 (s) 46.44 (s) 53.20 (s) 56.82 (s) 63.12 (s) 65.93 (s) 71.92 (s) 75.53 (s) 79.15 (s) 80.19 (s) 82.45 (s) 114.27 (s) 123.99 (s) 124.98 (s) 129.13 (s) 129.19 (s) 130.26 (s) 130.61 (s) 131.80 (s) 134.97 (s) 136.25 (s) 137.62 (s) 140.85 (s) 149.64 (s) 155.68 (s) 177.11 (s) 178.93 (s); HR-MS ESI/APCI Dual m/z: 668.3916 [M + H]⁺ (calcd for C₃₇H₅₃N₃O₈: 668.3905).

(1S)-1-{5-[(4-{(1E)-4-[(1-{[(2-Amino-2-oxoethyl)amino]methyl}cyclohexyl)amino]-3,3-dimethyl-4-oxobut-1-en-1-yl}-2-methylphenyl)methyl]-2-hydroxy-4-(propan-2-yl)phenyl}-1,5-anhydro-D-glucitol (28e)

Compound 24e (265 mg, 0.318 mmol) and compound 26 (46 mg, 0.413 mmol) were used as starting materials and synthesized in the same manner as compound 28b to give the title compound (13 mg, 5.6% for 2 steps) as a colorless amorphous. ¹H-NMR (600 MHz, CD₃OD) δ ppm 1.09 (d, J = 6.4 Hz, 6H), 1.26–1.40 (m, 14H), 1.48–1.61 (m, 3H), 2.11–2.16 (m, 2H), 2.32 (s, 3H), 2.85 (s, 2H), 2.88–2.95 (m, 1H), 3.25 (s, 2H), 3.37–3.41 (m, 2H), 3.46 (t, J = 8.9 Hz, 1H), 3.52–3.57 (m, 1H), 3.65–3.70 (m, 1H), 3.84 (d, J = 12.4 Hz, 1H), 3.89 (s, 2H), 4.47 (d, J = 9.6 Hz, 1H), 6.43 (d, J = 16.5 Hz, 1H), 6.58 (d, J = 16.5 Hz, 1H), 6.76 (d, J = 7.8 Hz, 1H), 6.80 (s, 1H), 6.97 (s, 1H), 7.12 (d, J = 7.8 Hz, 1H), 7.25 (s, 1H); ¹³C-NMR (126 MHz, CD₃OD) δ ppm 8.61 (s) 20.00 (s) 22.73 (s) 24.10 (s) 24.22 (s) 25.94 (s) 26.86 (s) 30.33 (s) 33.83 (s) 36.36 (s) 46.86 (s) 53.21 (s) 57.56 (s) 60.30 (s) 63.13 (s) 71.93 (s) 75.55 (s) 79.14 (s) 80.19 (s) 82.46 (s) 114.30 (s) 124.02 (s) 124.96 (s) 129.15 (s) 129.18 (s) 130.29 (s) 131.08 (s) 131.84 (s) 134.88 (s) 136.10 (s) 137.67 (s) 141.02 (s) 149.65 (s) 155.70 (s) 176.77 (s) 178.89 (s); HR-MS ESI/APCI Dual m/z: 682.4083[M + H]⁺ (calcd for C₃₈H₅₅N₃O₈: 682.4062).

(1S)-1-{5-[(4-{(1E)-4-[(3-{[(2-Amino-2-oxoethyl)amino]methyl}oxetan-3-yl)amino]-3,3-dimethyl-4-oxobut-1-en-1-yl}-2-methylphenyl)methyl]-2-hydroxy-4-(propan-2-yl)phenyl}-1,5-anhydro-D-glucitol (28f)

Sodium cyanoborohydride (27 mg) was added to a solution of compound 24f (117 mg, 0.145 mmol) and compound 26 (48 mg, 0.434 mmol) in methanol solution (4 mL). The reaction solution was stirred at the room temperature for 18 h. After the solvent was distilled off under reduced pressure, water was added and the mixture was extracted with ethyl acetate. The organic layer was washed with saturated aqueous sodium hydrogen carbonate solution and saturated brine, and dried over anhydrous magnesium sulfate. After the desiccant was filtered off, the solvent was distilled off under reduced pressure. The residue was purified by silica gel column chromatography (chloroform : methanol = 100 : 0 to 50 : 50) to obtain the crude intermediate (68 mg). The crude intermediate (68 mg) in methanol : triethylamine : water (5 : 1 : 1) solution (2 mL) was stirred at the room temperature for 18 h. After the solvent was distilled off under reduced pressure, the residue was purified by NH silica gel column chromatography (ethyl acetate : ethanol : water = 10 : 2 : 1) to obtain the title compound (16 mg, 17% in 2steps) as a colorless amorphous. ¹H-NMR (300 MHz, CD₃OD) δ ppm 1.10 (d, J = 6.7 Hz, 6H), 1.38 (s, 6H), 2.30 (s, 3H), 2.86–2.98 (m, 1H), 3.08 (s, 2H), 3.25 (s, 2H), 3.36–3.40 (m, 2H), 3.41–3.49 (m, 1H), 3.50–3.58 (m, 1H), 3.63–3.72 (m, 1H), 3.80–3.91 (m, 3H), 4.46 (d, J = 9.5 Hz, 1H), 4.50 (d, J = 7.0 Hz, 2H), 4.67 (d, J = 7.0 Hz, 2H), 6.38 (d, J = 16.3 Hz, 1H), 6.50 (d, J = 16.3 Hz, 1H), 6.76 (d, J = 7.9 Hz, 1H), 6.80 (s, 1H), 6.95 (s, 1H), 7.11 (d, J = 7.9 Hz, 1H), 7.24 (s, 1H); ¹³C-NMR (126 MHz, CD₃OD) δ ppm 19.97 (s) 24.10 (s) 24.21 (s) 25.86 (s) 25.98 (s) 30.31 (s) 36.32 (s) 45.85 (s) 46.66 (s) 51.91 (s) 53.29 (s) 53.36 (s) 54.68 (s) 58.09 (s) 61.85 (s) 63.12 (s) 63.78 (s) 71.91 (s) 75.53 (s) 79.14 (s) 80.16 (s) 80.19 (s) 82.44 (s) 114.26 (s) 123.98 (s) 125.08 (s) 125.11 (s) 129.17 (s) 129.21 (s) 130.25 (s) 130.43 (s) 131.10 (s) 131.76 (s) 134.46 (s) 134.53 (s) 136.28 (s) 137.58 (s) 137.62 (s) 140.79 (s) 140.89 (s) 149.62 (s) 155.67 (s) 177.26 (s) 178.90 (s) 179.89 (s); HR-MS ESI/APCI Dual m/z: 656.3561[M + H]⁺ (calcd for C₃₅H₄₉N₃O₉: 656.3542).

(1S)-1-[5-({4-[(1E)-4-{[3-{[(2-Amino-2-oxoethyl)amino]methyl}-1-(tert-butoxycarbonyl)azetidin-3-yl]amino}-3,3-dimethyl-4-oxobut-1-en-1-yl]-2-methylphenyl}methyl)-2-hydroxy-4-(propan-2-yl)phenyl]-1,5-anhydro-D-glucitol (28g)

Sodium triacetoxyborohydride (30 mg) was added to a solution of compound 24g (550 mg, 0.606 mmol) and compound 26 (87 mg, 0.788 mmol) and acetic acid (45 µL) in N,N-dimethylformamide solution (6 mL). The reaction solution was stirred at the room temperature for 18 h. Water was added and the mixture was extracted with chloroform. The organic layer was washed with saturated aqueous sodium hydrogen carbonate solution and dried over anhydrous magnesium sulfate. After the desiccant was filtered off, the solvent was distilled off under reduced pressure. The residue was purified by silica gel column chromatography (chloroform : methanol = 99 : 1 to 50 : 50) to obtain the crude intermediate (313 mg). The crude intermediate (163 mg) and 4.88 sodium methoxide (415 µL) in methanol solution (1.7 mL) was stirred at the room temperature for 18 h. After the solvent was distilled off under reduced pressure, the residue was purified by NH silica gel column chromatography (ethyl acetate : ethanol : water = 20 : 2 : 1 to 7 : 2 : 1) to obtain the title compound (80 mg, 34% in 2steps) as a colorless amorphous. ¹H-NMR (300 MHz, CD₃OD) δ ppm 1.10 (d, J = 6.84 Hz, 6H), 1.38 (s, 6H), 1.42 (s, 9H), 2.30 (s, 3H), 2.86–3.01 (m, 3H), 3.22 (s, 2H), 3.37–3.72 (m, 6H), 3.81–4.00 (m, 6H), 4.46 (d, J = 9.33 Hz, 1H), 6.28–6.42 (m, 1H), 6.44–6.55 (m, 1H), 6.75 (d, J = 7.93 Hz, 1H), 6.80 (s, 1H), 6.95 (s, 1H), 7.11 (dd, J = 7.93, 1.55 Hz, 1H), 7.23 (s, 1H); MS (ESI/APCI Dual) m/z: 755[M + H]⁺.

(1S)-1-{5-[(4-{(1E)-4-[(3-{[(2-Amino-2-oxoethyl)amino]methyl}azetidin-3-yl)amino]-3,3-dimethyl-4-oxobut-1-en-1-yl}-2-methylphenyl)methyl]-2-hydroxy-4-(propan-2-yl)phenyl}-1,5-anhydro-D-glucitol (29g)

The compound 28g (80 mg, 0.106 mmol) in trifluoroacetic acid solution (1.0 mL) was stirred at the room temperature for 18 h. After the solvent was distilled off under reduced pressure, the residue was purified by NH silica gel column chromatography (ethyl acetate : ethanol : water = 20 : 2 : 1 to 7 : 2 : 1) to obtain the title compound (44 mg, 63%) as a colorless amorphous. ¹H-NMR (500 MHz, CD₃OD) δ ppm 1.04–1.11 (m, 6H), 1.35–1.39 (m, 3H), 1.46 (s, 3H), 2.31 (s, 3H), 2.88–2.95 (m, 1H), 3.20–3.94 (m, 12H), 4.46 (d, J = 9.56 Hz, 1H), 6.27–6.41 (m, 1H), 6.51 (br dd, J = 18.73, 16.44 Hz, 1H), 6.76 (br d, J = 8.03 Hz, 1H), 6.80 (s, 1H), 6.95 (s, 1H), 7.11 (br d, J = 7.64 Hz, 1H), 7.24 (s, 1H); ¹³C-NMR (126 MHz, CD₃OD) δ ppm 19.96 (s) 24.09 (s) 24.21 (s) 25.84 (s) 26.42 (s) 26.48 (s) 30.31 (s) 36.31 (s) 40.19 (s) 53.36 (s) 54.95 (s) 55.01 (s) 55.61 (s) 56.80 (s) 57.80 (s) 63.13 (s) 66.34 (s) 71.92 (s) 71.96 (s) 75.55 (s) 75.73 (s) 79.11 (s) 80.19 (s) 82.44 (s) 114.28 (s) 124.02 (s) 125.06 (s) 125.17 (s) 129.13 (s) 129.16 (s) 129.25 (s) 130.19 (s) 130.24 (s) 130.38 (s) 131.76 (s) 133.48 (s) 137.61 (s) 141.09 (s) 149.61 (s) 155.70 (s) 155.74 (s); HR-MS ESI/APCI Dual m/z: 655.3718[M + H]⁺ (calcd for C₃₅H₅₀N₄O₈: 655.3701).

(1S)-2,3,4,6-Tetra-O-acetyl-1,5-anhydro-1-{5-[(4-bromo-2-methylphenyl)methyl]-2-methoxy-4-(propan-2-yl)phenyl}-D-glucitol (30)

Methyl iodide (33 mL, 0.530 mol) in N,N-dimethylformamide (50 mL) was added to a suspension of compound 11 (392 g, 0.506 mol) and potassium carbonate (73.4 g, 0.531 mol) in N,N-dimethylformamide (0.95 L). The reaction solution was stirred for 1 h, and after adding potassium carbonate (70.0 g, 0.506 mol) and methyl iodide (31.5 mL, 0.506 mol), the mixture was stirred for 1 h. Potassium carbonate (70.0 g, 0.506 mol) and methyl iodide (31.5 mL, 0.506 mol) were added again, and the mixture was stirred for 1 h. Methyl iodide (15.8 mL, 0.254 mol) was added, and the mixture was stirred overnight at room temperature. After stirring at 50°C for 2 h, the mixture was diluted with toluene (1.25 L), and water (1.0 L) was added. The two layers were separated, and the organic layer was washed twice with water (1.0 L) and 10% brine (1.0 L), and then concentrated under reduced pressure. The residue was dissolved with isopropyl alcohol (350 mL) at 40°C, and the solution was cooled to room temperature and stirred overnight. The resulting precipitate was filtered off and dried to give the title compound (155 g, 46%) in the form of a white powder. ¹H-NMR (300 MHz, CDCl₃) δ ppm 1.12 (d, J = 6.8 Hz, 3H), 1.13 (d, J = 6.8 Hz, 3H), 1.55 (s, 3H), 1.75 (s, 3H), 1.99 (s, 3H), 2.04 (s, 3H), 2.05 (s, 3H), 2.29 (s, 3H), 2.83–2.96 (m, 1H), 3.86 (s, 3H), 4.08–4.17 (m, 1H), 4.18–4.28 (m, 1H), 4.78–4.89 (m, 1H), 5.13–5.23 (m, 1H), 5.27–5.35 (m, 2H), 6.51–6.57 (m, 1H), 6.80 (s, 1H), 6.96 (s, 1H), 7.12–7.20 (m, 1H), 7.30 (s, 1H).

(1S)-2,3,4,6-Tetra-O-acetyl-1,5-anhydro-1-{5-[(4-bromo-2-methylphenyl)methyl]-2-ethoxy-4-(propan-2-yl)phenyl}-D-glucitol (34b)

The compound 12 (100 g) in methanol : triethylamine : water (5 : 1 : 1) solution (12 L) was stirred at the room temperature for 3 d. The reaction solution was concentrated, and dissolved in 2M hydrochloric acid and extracted twice with chloroform. The organic layers were combined, washed with saturated brine, and dried over anhydrous magnesium sulfate. After the desiccant was filtered off, the solvent was distilled off under reduced pressure to obtain the intermediate 31 (80.4 g) as a light brown amorphous. To a suspension of the intermediate 31 (1.00 g, 2.08 mmol) and potassium carbonate (862 mg, 6.24 mmol) in N,N-dimethylformamide (6.0 mL) was added to ethyl iodide (500 µL, 6.24 mmol). The reaction mixture was stirred at 50°C for 8 h. Water was added to the reaction solution and extracted with chloroform. The organic layer was washed with saturated brine, dried over anhydrous magnesium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (methanol : chloroform = 0 : 100 to 10 : 90) to obtain the intermediate (950 mg, 90%) as a colorless liquid. A solution of the intermediate (867 mg, 1.70 mmol) and acetic anhydride (4.3 mL) in pyridine (5.2 mL) was stirred at room temperature for 3 d. The reaction solution was added to ice water and extracted with toluene. The organic layer was washed with 2M hydrochloric acid and saturated brine, and dried over sodium sulfate. After the desiccant was filtered off, the solvent was evaporated under reduced pressure, and the residue was purified by silica gel column chromatography (hexane : ethyl acetate = 85 : 15 → 65 : 35) to give the title compound (1.00 g, 90% in 3steps). ¹H-NMR (300 MHz, CDCl₃) δ ppm 1.07–1.15 (m, 6H), 1.46 (t, J = 6.9 Hz, 3H), 1.75 (s, 3H), 2.00 (s, 3H), 2.04 (s, 3H), 2.05 (s, 3H), 2.28 (s, 3H), 2.83–2.94 (m, 1H), 3.72–3.90 (m, 3H), 4.02–4.15 (m, 3H), 4.19–4.28 (m, 1H), 4.76–4.89 (m, 1H), 5.15–5.22 (m, 1H), 5.27–5.38 (m, 2H), 6.54 (d, J = 8.2 Hz, 1H), 6.79 (s, 1H), 6.94 (s, 1H), 7.16 (dd, J = 8.2, 1.7 Hz, 1H), 7.31 (d, J = 1.7 Hz, 1H); MS (ESI/APCI Dual) m/z: 699 [M + Na]⁺.

(1S)-2,3,4,6-Tetra-O-acetyl-1-{2-[2-(acetyloxy)ethoxy]-5-[(4-bromo-2-methylphenyl)methyl]-4-(propan-2-yl)phenyl}-1,5-anhydro-D-glucitol (34c)

To a suspension of Intermediate 31 (1.00 g, 2.08 mmol) and potassium carbonate (861 mg, 6.23 mmol) in N, N-dimethylformamide (5 mL) was added to 2-iodoethanol (485 µL, 6.23 mmol). The reaction mixture stirred at 150°C. for 3 h. Water was added to the reaction mixture and extracted twice with ethyl acetate. The organic layers were combined, washed with 10% brine and water, and dried over anhydrous magnesium sulfate. The desiccant was filtered off and concentrated under reduced pressure. The residue was purified by silica gel chromatography (ethyl acetate only → ethyl acetate : ethanol : water = 15 : 2 : 1) to give the intermediate (370 mg) as a colorless amorphous. Acetic anhydride (900 µL) was added to a solution of the intermediate (360 mg, 0.685 mmol) in pyridine (1.08 mL), and the mixture was stirred overnight at room temperature. The reaction solution was added to ice water and extracted with ethyl acetate. The organic layer was washed with 2M hydrochloric acid twice and saturated brine, and then dried over anhydrous magnesium sulfate. After the desiccant was filtered off, the solvent was evaporated under reduced pressure to give the title compound (480 mg, 37% in 3steps) as a colorless amorphous. ¹H-NMR (300 MHz, CDCl₃) δ ppm 1.07–1.15 (m, 6H), 1.74 (s, 3H), 1.99 (s, 3H), 2.04 (s, 3H), 2.06 (s, 3H), 2.17 (s, 3H), 2.28 (s, 3H), 2.88–2.94 (m, 1H), 3.72–3.92 (m, 3H), 4.04–4.33 (m, 4H), 4.36–4.49 (m, 1H), 4.52–4.64 (m, 1H), 4.75 (br s, 1H), 5.19 (t, J = 9.2 Hz, 1H), 5.30 (t, J = 9.2 Hz, 1H), 5.41 (br s, 1H), 6.53 (d, J = 8.2 Hz, 1H), 6.79 (s, 1H), 6.93 (s, 1H), 7.18 (d, J = 8.2 Hz, 1H), 7.32 (s, 1H); MS (ESI/APCI Dual) m/z: 757 [M + Na]⁺.

(1S)-2,3,4,6-Tetra-O-acetyl-1,5-anhydro-1-{5-[(4-bromo-2-methylphenyl)methyl]-2-[2-(dimethylamino)ethoxy]-4-(propan-2-yl)phenyl}-D-glucitol (34d)

Intermediate 31 (2.0 g, 4.15 mmol) and 2-iodo-N,N-dimethylethan-1-amine (2.07 g, 10.4 mmol) were used as starting materials and synthesized in the same manner as compound 34c to give the title compound (520 mg, 17% for 2 steps) as a brown amorphous. ¹H-NMR (300 MHz, CDCl₃) δ ppm 1.12 (m, 6H), 1.75 (s, 3H), 1.99 (s, 3H), 2.04 (s, 3H), 2.05 (s, 3H), 2.08 (s, 3H), 2.28 (s, 3H), 2.49 (s, 6H), 2.84–3.00 (m, 3H), 3.76–3.91 (m, 3H), 4.02–4.26 (m, 4H), 4.78–4.94 (m, 1H), 5.13–5.37 (m, 3H), 6.53 (d, J = 8.08 Hz, 1H), 6.81 (s, 1H), 6.94 (s, 1H), 7.17 (dd, J = 8.00, 1.94 Hz, 1H), 7.32 (d, J = 2.18 Hz, 1H); MS (ESI/APCI Dual) m/z: 720 [M + H]⁺.

(1S)-2,3,4,6-Tetra-O-acetyl-1,5-anhydro-1-[5-({4-[(1E)-3-carboxy-3-methylbut-1-en-1-yl]-2-methylphenyl}methyl)-2-methoxy-4-(propan-2-yl)phenyl]-D-glucitol (35a)

In an argon atmosphere, an acetonitrile (200 mL) suspension of compound 30 (10.0 g, 15.1 mmol), compound 17 (4.12 g, 36.1 mmol), palladium (II) acetate (337 mg, 1.51 mmol), tri-o-tolylphosphine (917 mg, 3.02 mmol), and triethylamine (10.5 mL, 75.5 mmol) was refluxed for 3 h. After cooling to room temperature, the reaction mixture was concentrated, the residue was dissolved in ethyl acetate (800 mL), and the solution was washed with 10% hydrochloric acid (200 mL), then dried over anhydrous magnesium sulfate. The filtrate was concentrated under reduced pressure, and the resulting residue was purified by performing neutral silica gel column chromatography (hexane: ethyl acetate = 8 : 2 to1 : 9) to obtain the title compound (10.4 g, 99%) in the form of a colorless amorphous. ¹H-NMR (600 MHz, CDCl₃) δ ppm 1.12 (d, J = 6.9 Hz, 3H), 1.13 (d, J = 6.9 Hz, 3H), 1.43 (s, 6H), 1.75 (s, 3H), 1.99 (s, 3H), 2.01–2.06 (m, 6H), 2.29 (s, 3H), 2.90–2.98 (m, 1H), 3.77–3.81 (m, 1H), 3.81–3.91 (m, 5H), 4.08–4.13 (m, 1H), 4.21 (dd, J = 12.2, 4.4 Hz, 1H), 4.79 (br d, J = 8.3 Hz, 1H), 5.17 (t, J = 9.6 Hz, 1H), 5.27–5.36 (m, 2H), 6.35 (d, J = 16.0 Hz, 1H), 6.43 (d, J = 16.0 Hz, 1H), 6.64 (d, J = 8.3 Hz, 1H), 6.80 (s, 1H), 6.95 (s, 1H), 7.06 (d, J = 8.3 Hz, 1H), 7.21 (s, 1H); MS (ESI/APCI Dual) m/z: 719 [M + Na]⁺.

(1S)-2,3,4,6-Tetra-O-acetyl-1,5-anhydro-1-{5-[(4-{(1E)-3,3-dimethyl-4-[(2-methyl-1-oxopropan-2-yl)amino]-4-oxobut-1-en-1-yl}-2-methylphenyl)methyl]-2-methoxy-4-(propan-2-yl)phenyl}-D-glucitol (36a)

Compound 22 (1.72 g, 19.4 mmol), water soluble carbodiimide hydrochloride (3.21 g, 16.8 mmol) and 1-hydroxy-1H-benzotriazole hydrate (2.27 g, 16.8 mmol) were added to a N,N-dimethylformamide (100 mL) solution of compound 35a (9.0 g, 12.9 mmol) in a nitrogen atmosphere, and the mixture was stirred for 16 h at room temperature. Water was added to the reaction liquid, and after extracting the mixture with ethyl acetate, the organic layer was washed with 10% brine, then dried over anhydrous magnesium sulfate. The desiccant was filtered out, and the solvent was distilled off under reduced pressure. The resulting residue was purified by performing silica gel column chromatography (hexane : ethyl acetate = 2 : 8 to 8 : 2) to obtain the colorless powder (8.07 g). Dess–Martin periodinane (6.69 g, 15.8 mmol) was added to the colorless powder (8.07 g, 10.5 mmol) in a nitrogen atmosphere, and the mixture was stirred for 1 h at room temperature. The resection mixture was distilled off under reduced pressure, and the resulting residue was purified by silica gel column chromatography (hexane : ethyl acetate ratio = 2 : 8 to 8 : 2) to obtain the title compound (7.6 g, 77%) in the form of a yellow amorphous. ¹H-NMR (600 MHz, CDCl₃) δ ppm 1.09–1.18 (m, 6H), 1.32 (s, 6H), 1.38 (s, 6H), 1.77 (s, 3H), 1.98 (s, 3H), 2.00–2.07 (m, 6H), 2.33 (s, 3H), 2.95 (spt, J = 6.8 Hz, 1H), 3.77–3.94 (m, 6H), 4.08–4.15 (m, 1H), 4.22 (dd, J = 11.9, 4.6 Hz, 1H), 4.82 (br s., 1H), 5.14–5.20 (m, 1H), 5.27–5.35 (m, 2H), 6.11 (br s, 1H), 6.31 (d, J = 16.5 Hz, 1H), 6.52 (d, J = 16.5 Hz, 1H), 6.67 (d, J = 7.8 Hz, 1H), 6.81 (s, 1H), 6.99 (s, 1H), 7.10 (d, J = 7.8 Hz, 1H), 7.24 (s, 1H), 9.33 (s, 1H); MS (ESI/APCI Dual) m/z: 766 [M + H]⁺, 788 [M + Na]⁺.

(1S)-2,3,4,6-Tetra-O-acetyl-1,5-anhydro-1-{5-[(4-{(1E)-3,3-dimethyl-4-[(2-methyl-1-oxopropan-2-yl)amino]-4-oxobut-1-en-1-yl}-2-methylphenyl)methyl]-2-ethoxy-4-(propan-2-yl)phenyl}-D-glucitol (36b)

Compound 34b (1.0 g, 1.48 mmol) was used as starting materials and synthesized in the same manner as compound 35a and 36a to give the title compound (175 mg, 15% for 3 steps) as a colorless amorphous. ¹H-NMR (300 MHz, CDCl₃) δ ppm 1.06–1.18 (m, 6H), 1.32 (s, 6H), 1.38 (s, 6H), 1.47 (t, J = 6.9 Hz, 3H), 1.77 (s, 3H), 1.99 (s, 3H), 2.04 (s, 3H), 2.04 (s, 3H), 2.33 (s, 3H), 2.85–3.03 (m, 1H), 3.72–3.97 (m, 3H), 4.01–4.16 (m, 3H), 4.17–4.28 (m, 1H), 4.72–4.91 (m, 1H), 5.11–5.21 (m, 1H), 5.27–5.37 (m, 2H), 6.11 (s, 1H), 6.26–6.35 (m, 1H), 6.48–6.57 (m, 1H), 6.67 (d, J = 7.9 Hz, 1H), 6.80 (s, 1H), 6.97 (s, 1H), 7.10 (d, J = 7.9 Hz, 1H), 7.25 (s, 1H), 9.33 (s, 1H); MS ESI/APCI Dual m/z: 780 [M + H]⁺, 802 [M + Na]⁺, 778 [M−H]⁻.

(1S)-2,3,4,6-Tetra-O-acetyl-1-{2-[2-(acetyloxy)ethoxy]-5-[(4-{(1E)-3,3-dimethyl-4-[(2-methyl-1-oxopropan-2-yl)amino]-4-oxobut-1-en-1-yl}-2-methylphenyl)methyl]-4-(propan-2-yl)phenyl}-1,5-anhydro-D-glucitol (36c)

Compound 34c (480 mg, 0.653 mmol) was used as starting materials and synthesized in the same manner as compound 35a and 36a to give the title compound (250 mg, 46% for 3 steps) as a colorless amorphous. ¹H-NMR (300 MHz, CDCl₃) δ ppm 1.13 (d, J = 6.8 Hz, 3H), 1.14 (d, J = 6.8 Hz, 3H), 1.33 (s, 6H), 1.39 (s, 6H), 1.76 (s, 3H), 1.98 (s, 3H), 2.04 (s, 3H), 2.05 (s, 3H), 2.18 (s, 3H), 2.32 (s, 3H), 2.88–3.01 (m, 1H), 3.76–3.91 (m, 3H), 4.05–4.13 (m, 1H), 4.15–4.30 (m, 3H), 4.37–4.48 (m, 1H), 4.53–4.63 (m, 1H), 4.73 (br s., 1H), 5.18 (t, J = 9.2 Hz, 1H), 5.30 (t, J = 9.2 Hz, 1H), 5.41 (br s., 1H), 6.11 (s, 1H), 6.34 (d, J = 16.0 Hz, 1H), 6.55 (d, J = 16.0 Hz, 1H), 6.67 (d, J = 7.8 Hz, 1H), 6.80 (s, 1H), 6.96 (s, 1H), 7.11 (d, J = 7.8 Hz, 1H), 7.25 (s, 1H), 9.33 (s, 1H); MS (ESI/APCI Dual) m/z: 838 [M + H]⁺, 860 [M + Na]⁺.

(1S)-2,3,4,6-Tetra-O-acetyl-1,5-anhydro-1-{2-[2-(dimethylamino)ethoxy]-5-[(4-{(1E)-3,3-dimethyl-4-[(2-methyl-1-oxopropan-2-yl)amino]-4-oxobut-1-en-1-yl}-2-methylphenyl)methyl]-4-(propan-2-yl)phenyl}-D-glucitol (36d)

Compound 34d (510 mg, 0.708 mmol) was used as starting materials and synthesized in the same manner as compound 35a and 36a to give the title compound (215 mg, 38% for 3 steps) as a pale orange amorphous. ¹H-NMR (300 MHz, CDCl₃) δ ppm 1.13 (m, 6H), 1.32 (s, 6H), 1.39 (s, 6H), 1.77 (s, 3H), 1.99 (s, 3H), 2.04 (m, 6H), 2.33 (s, 3H), 2.42 (s, 6H), 2.79–2.99 (m, 3H), 3.76–3.90 (m, 3H), 4.06–4.26 (m, 4H),4.75–4.88 (m, 1H), 5.11–5.42 (m, 3H), 6.13 (s, 1H), 6.26–6.36 (m, 1H), 6.48–6.56 (m, 1H), 6.66 (d, J = 7.93 Hz, 1H), 6.83 (s, 1H), 6.98 (s, 1H), 7.10 (dd, J = 7.85, 1.63 Hz, 1H), 7.25 (s, 1H), 9.34 (s, 1H); MS (ESI/APCI Dual) m/z: 823 [M + H]⁺.

(1S)-1-[5-({4-[(1E)-4-({1-[(2-Amino-2-oxoethyl)amino]-2-methylpropan-2-yl}amino)-3,3-dimethyl-4-oxobut-1-en-1-yl]-2-methylphenyl}methyl)-2-methoxy-4-(propan-2-yl)phenyl]-1,5-anhydro-D-glucitol (38a)

Compound 36a (300 mg, 0.39 mmol) was used as starting materials and synthesized in the same manner as compound 28a to give the title compound (41 mg, 16% for 2 steps) as a colorless amorphous. ¹H-NMR (600 MHz, CD₃OD) δ ppm 1.12–1.17 (m, 6H), 1.30 (s, 6H), 1.35 (s, 6H), 2.32 (s, 3H), 2.66 (s, 2H), 2.99 (s, 1H), 3.18 (s, 2H), 3.34 (d, J = 2.3 Hz, 2H), 3.43–3.48 (m, 1H), 3.49–3.53 (m, 1H), 3.59–3.64 (m, 1H), 3.80–3.84 (m, 1H), 3.84 (s, 3H), 3.92 (s, 2H), 4.59–4.63 (m, 1H), 6.35–6.39 (m, 1H), 6.48–6.52 (m, 1H), 6.74 (d, J = 7.8 Hz, 1H), 6.92 (s, 1H), 7.07 (s, 1H), 7.09–7.12 (m, 1H), 7.24 (s, 1H); ¹³C-NMR (126 MHz, S CD₃OD) δ ppm 20.00 (s) 24.05 (s) 24.19 (s) 25.30 (s) 25.92 (s) 30.73 (s) 36.36 (s) 46.57 (s) 53.24 (s) 54.69 (s) 56.52 (s) 59.61 (s) 63.43 (s) 72.31 (s) 75.75 (s) 76.96 (s) 80.29 (s) 82.54 (s) 109.64 (s) 125.00 (s) 126.26 (s) 129.16 (s) 130.18 (s) 130.20 (s) 130.58 (s) 131.75 (s) 134.99 (s) 136.31 (s) 137.61 (s) 140.64 (s) 149.79 (s) 158.61 (s) 177.21 (s) 178.81 (s); HR-MS ESI/APCI Dual m/z: 656.3951 [M + H]⁺ (calcd for C₃₆H₅₃N₃O₈: 656.3905).

(1S)-1-[5-({4-[(1E)-4-({1-[(2-Amino-2-oxoethyl)amino]-2-methylpropan-2-yl}amino)-3,3-dimethyl-4-oxobut-1-en-1-yl]-2-methylphenyl}methyl)-2-ethoxy-4-(propan-2-yl)phenyl]-1,5-anhydro-D-glucitol (38b)

Compound 36b (175 mg, 0.224 mmol) was used as starting materials and synthesized in the same manner as compound 28g to give the title compound (80 mg, 56% for 2 steps) as a colorless amorphous. ¹H-NMR (600 MHz, CD₃OD) δ ppm 1.10–1.16 (m, 6H), 1.30 (s, 6H), 1.35 (s, 6H), 1.41 (t, J = 6.9 Hz, 3H), 2.32 (s, 3H), 2.66 (s, 2H), 2.95–3.02 (m, 1H), 3.18 (s, 2H), 3.33–3.38 (m, 2H), 3.45 (t, J = 8.7 Hz, 1H), 3.55–3.64 (m, 2H), 3.83 (dd, J = 11.9, 1.8 Hz, 1H), 3.92 (s, 2H), 4.05–4.13 (m, 2H), 4.59 (d, J = 9.6 Hz, 1H), 6.37 (d, J = 16.5 Hz, 1H), 6.50 (d, J = 16.5 Hz, 1H), 6.75 (d, J = 7.8 Hz, 1H), 6.91 (s, 1H), 7.05 (s, 1H), 7.11 (d, J = 7.8 Hz, 1H), 7.24 (s, 1H); ¹³C-NMR (126 MHz, CD₃OD) δ ppm 15.49 (s) 19.99 (s) 24.06 (s) 24.18 (s) 25.30 (s) 25.92 (s) 30.66 (s) 36.36 (s) 46.57 (s) 53.24 (s) 54.69 (s) 59.60 (s) 63.46 (s) 65.80 (s) 72.30 (s) 75.36 (s) 77.36 (s) 80.33 (s) 82.51 (s) 111.13 (s) 125.00 (s) 126.42 (s) 129.16 (s) 130.21 (s) 130.24 (s) 130.57 (s) 131.88 (s) 134.99 (s) 136.31 (s) 137.62 (s) 140.61 (s) 149.76 (s) 158.05 (s) 177.22 (s) 178.79 (s); HR-MS ESI/APCI Dual m/z: 670.4090 [M + H]⁺ (calcd for C₃₇H₅₅N₃O₈: 670.4062).

(1S)-1-[5-({4-[(1E)-4-({1-[(2-Amino-2-oxoethyl)amino]-2-methylpropan-2-yl}amino)-3,3-dimethyl-4-oxobut-1-en-1-yl]-2-methylphenyl}methyl)-2-(2-hydroxyethoxy)-4-(propan-2-yl)phenyl]-1,5-anhydro-D-glucitol (38c)

Compound 36c (120 mg, 0.143 mmol) was used as starting materials and synthesized in the same manner as compound 28g to give the title compound (30 mg, 30% for 2 steps) as a colorless amorphous. ¹H-NMR (600 MHz, CD₃OD) δ ppm 1.13 (d, J = 6.9 Hz, 3H), 1.14 (d, J = 6.9 Hz, 3H), 1.30 (s, 6H), 1.34 (s, 6H), 2.32 (s, 3H), 2.66 (s, 2H), 2.98 (m, 1H), 3.17 (s, 2H), 3.34 (s, 2H), 3.37–3.40 (m, 1H), 3.45–3.51 (m, 2H), 3.60–3.65 (m, 1H), 3.81–3.90 (m, 3H), 3.91 (s, 2H), 4.08–4.15 (m, 2H), 4.68 (d, J = 9.2 Hz, 1H), 6.37 (d, J = 16.5 Hz, 1H), 6.50 (d, J = 16.5 Hz, 1H), 6.73 (d, J = 7.8 Hz, 1H), 6.93 (s, 1H), 7.07–7.12 (m, 2H), 7.24 (s, 1H); ¹³C-NMR (126 MHz, CD₃OD) δ ppm 20.00 (s) 24.04 (s) 24.20 (s) 25.30 (s) 25.92 (s) 30.68 (s) 36.38 (s) 46.57 (s) 53.24 (s) 54.69 (s) 59.60 (s) 62.07 (s) 63.39 (s) 72.00 (s) 72.27 (s) 76.17 (s) 77.04 (s) 80.28 (s) 82.47 (s) 111.41 (s) 125.00 (s) 127.04 (s) 129.15 (s) 130.18 (s) 130.56 (s) 130.70 (s) 131.75 (s) 135.00 (s) 136.32 (s) 137.61 (s) 140.58 (s) 149.75 (s) 157.77 (s) 177.23 (s) 178.78 (s); HR-MS ESI/APCI Dual m/z: 686.3997 [M + H]⁺ (calcd for C₃₇H₅₅N₃O₉: 686.4011).

(1S)-1-[5-({4-[(1E)-4-({1-[(2-Amino-2-oxoethyl)amino]-2-methylpropan-2-yl}amino)-3,3-dimethyl-4-oxobut-1-en-1-yl]-2-methylphenyl}methyl)-2-[2-(dimethylamino)ethoxy]-4-(propan-2-yl)phenyl]-1,5-anhydro-D-glucitol (38d)

Compound 36c (120 mg, 0.146 mmol) was used as starting materials and synthesized in the same manner as compound 28b to give the title compound (35 mg, 27% for 2 steps) as a colorless amorphous. ¹H-NMR (500 MHz, CD₃OD) δ ppm 1.14 (dd, J = 6.50, 5.35 Hz, 6H), 1.30 (s, 6H), 1.35 (s, 6H), 2.32 (s, 3H), 2.40 (s, 6H), 2.66 (s, 2H), 2.74–2.82 (m, 1H), 2.85–2.92 (m, 1H), 2.95–3.06 (m, 1H), 3.17 (s, 2H), 3.32–3.39 (m, 2H), 3.42–3.49 (m, 2H), 3.58–3.65 (m, 1H), 3.82 (dd, J = 11.85, 1.91 Hz, 1H), 3.92 (s, 2H), 4.08–4.23 (m, 2H), 4.60–4.67 (m, 1H), 6.33–6.41 (m, 1H), 6.47–6.55 (m, 1H), 6.73 (d, J = 8.03 Hz, 1H), 6.93 (s, 1H), 7.05–7.13 (m, 2H), 7.24 (d, J = 1.15 Hz, 1H); ¹³C-NMR (126 MHz, CD₃OD) δ ppm 20.00 (s) 24.04 (s) 24.18 (s) 25.30 (s) 25.92 (s) 30.70 (s) 36.37 (s) 46.07 (s) 46.57 (s) 53.24 (s) 54.69 (s) 59.59 (s) 63.44 (s) 67.75 (s) 72.37 (s) 76.03 (s) 77.09 (s) 80.41 (s) 82.59 (s) 111.02 (s) 125.00 (s) 126.80 (s) 129.16 (s) 130.18 (s) 130.56 (s) 130.70 (s) 131.77 (s) 135.00 (s) 136.32 (s) 137.60 (s) 140.57 (s) 149.78 (s) 157.67 (s) 177.22 (s) 178.78 (s); HR-MS ESI/APCI Dual m/z: 713.4498 [M + H]⁺ (calcd for C₃₉H₆₀N₄O₈: 713.4484).

(1S)-1-[5-({4-[(1E)-4-({1-[(4-Amino-4-oxobutyl)amino]-2-methylpropan-2-yl}amino)-3,3-dimethyl-4-oxobut-1-en-1-yl]-2-methylphenyl}methyl)-2-methoxy-4-(propan-2-yl)phenyl]-1,5-anhydro-D-glucitol (41b)

Compound 36a (360 mg, 0.470 mmol) and 4-aminobutanamide (57 mg, 0.558 mmol) was used as starting materials and synthesized in the same manner as compound 28a to give the title compound (151 mg, 47% for 2 steps) as a colorless amorphous. ¹H-NMR (600 MHz, CD₃OD) δ ppm 1.14 (d, J = 6.7 Hz, 3H), 1.15 (d, J = 6.7 Hz, 3H), 1.29–1.32 (m, 6H), 1.33–1.36 (m, 6H), 1.61–1.70 (m, 2H), 2.11 (t, J = 7.3 Hz, 2H), 2.31–2.35 (m, 3H), 2.61 (t, J = 6.9 Hz, 2H), 2.72 (br s., 2H), 2.93–3.02 (m, 1H), 3.37–3.42 (m, 2H), 3.44–3.49 (m, 1H), 3.50–3.55 (m, 1H), 3.60–3.65 (m, 1H), 3.79–3.86 (m, 4H), 3.92 (s, 2H), 4.62 (d, J = 9.6 Hz, 1H), 6.36 (d, J = 16.0 Hz, 1H), 6.50 (d, J = 16.0 Hz, 1H), 6.73 (d, J = 7.8 Hz, 1H), 6.92 (s, 1H), 7.08–7.12 (m, 2H), 7.24 (s, 1H); ¹³C-NMR (126 MHz, CD₃OD) δ ppm 20.03 (s) 24.05 (s) 24.21 (s) 25.37 (s) 25.87 (s) 30.74 (s) 34.27 (s) 36.37 (s) 46.54 (s) 50.72 (s) 54.31 (s) 56.52 (s) 59.44 (s) 63.40 (s) 72.29 (s) 75.72 (s) 76.99 (s) 80.28 (s) 82.54 (s) 109.65 (s) 125.02 (s) 126.24 (s) 129.18 (s) 130.13 (s) 130.16 (s) 130.68 (s) 131.83 (s) 134.69 (s) 136.23 (s) 137.59 (s) 140.69 (s) 149.82 (s) 158.62 (s) 178.68 (s) 179.14 (s); HR-MS ESI/APCI Dual m/z: 684.4246 [M + H]⁺ (calcd for C₃₈H₅₇N₃O₈: 684.4218).

(1S)-1-[5-({4-[(1E)-4-({1-[(6-Amino-6-oxohexyl)amino]-2-methylpropan-2-yl}amino)-3,3-dimethyl-4-oxobut-1-en-1-yl]-2-methylphenyl}methyl)-2-methoxy-4-(propan-2-yl)phenyl]-1,5-anhydro-D-glucitol (41c)

Compound 36a (300 mg, 0.392 mmol) and 5-aminopentanamide (76.5 mg, 0.588 mmol) was used as starting materials and synthesized in the same manner as compound 28b to give the title compound (141 mg, 52% for 2 steps) as a colorless amorphous. ¹H-NMR (600 MHz, CD₃OD) δ ppm 1.14 (t, J = 6.9 Hz, 6H), 1.17–1.24 (m, 2H), 1.26–1.37 (m, 14H), 1.45–1.53 (m, 2H), 2.12 (t, J = 7.6 Hz, 2H), 2.33 (s, 3H), 2.49 (t, J = 7.3 Hz, 2H), 2.61 (s, 2H), 2.92–3.00 (m, 1H), 3.33–3.38 (m, 2H), 3.46 (t, J = 8.5 Hz, 1H), 3.51–3.56 (m, 1H), 3.62 (dd, J = 11.9, 5.0 Hz, 1H), 3.79–3.87 (m, 4H), 3.92 (s, 2H), 4.62 (d, J = 9.6 Hz, 1H), 6.34 (d, J = 16.0 Hz, 1H), 6.49 (d, J = 16.0 Hz, 1H), 6.73 (d, J = 8.3 Hz, 1H), 6.92 (s, 1H), 7.06–7.12 (m, 2H), 7.24 (s, 1H); ¹³C-NMR (126 MHz, CD₃OD) δ ppm 20.08 (s) 24.08 (s) 24.22 (s) 25.42 (s) 25.90 (s) 26.80 (s) 28.06 (s) 30.51 (s) 30.74 (s) 36.44 (s) 36.56 (s) 46.55 (s) 51.37 (s) 54.39 (s) 56.52 (s) 59.90 (s) 63.42 (s) 72.29 (s) 75.73 (s) 76.99 (s) 80.30 (s) 82.54 (s) 109.66 (s) 125.05 (s) 126.25 (s) 129.15 (s) 130.07 (s) 130.15 (s) 130.64 (s) 131.90 (s) 134.76 (s) 136.25 (s) 137.53 (s) 140.70 (s) 149.83 (s) 158.63 (s) 178.79 (s) 179.18 (s); HR-MS ESI/APCI Dual m/z: 712.4554 [M + H]⁺ (calcd for C₄₀H₆₁N₃O₈: 712.4531).

(1S)-1,5-Anhydro-1-{5-[(4-{(1E)-4-[(1-{[2-(dimethylamino)ethyl]amino}-2-methylpropan-2-yl)amino]-3,3-dimethyl-4-oxobut-1-en-1-yl}-2-methylphenyl)methyl]-2-methoxy-4-(propan-2-yl)phenyl}-D-glucitol (41d)

Compound 36a (150 mg, 0.196 mmol) and N¹,N¹-dimethylethane-1,2-diamine (26 mg, 0.294 mmol) was used as starting materials and synthesized in the same manner as compound 28b to give the title compound (77.1 mg, 61% for 2 steps) as a colorless amorphous. ¹H-NMR (600 MHz, c 1.15 (d, J = 6.4 Hz, 6H), 1.30 (s, 6H), 1.33 (s, 6H), 2.10 (s, 6H), 2.20 (t, J = 6.4 Hz, 2H), 2.34 (s, 3H), 2.58–2.63 (m, 4H), 2.94–3.02 (m, 1H), 3.34–3.39 (m, 2H), 3.43–3.49 (m, 1H), 3.48–3.54 (m, 1H), 3.59–3.66 (m, 1H), 3.80–3.88 (m, 4H), 3.92 (s, 2H), 4.61 (d, J = 9.6 Hz, 1H), 6.35 (d, J = 16.0 Hz, 1H), 6.50 (d, J = 16.0 Hz, 1H), 6.73 (d, J = 7.8 Hz, 1H), 6.93 (s, 1H), 7.08 (s, 1H), 7.11 (d, J = 7.8 Hz, 1H), 7.26 (s, 1H); ¹³C-NMR (126 MHz, v 20.07 (s) 24.08 (s) 24.24 (s) 25.28 (s) 25.87 (s) 30.75 (s) 36.39 (s) 45.74 (s) 46.60 (s) 54.29 (s) 56.52 (s) 60.01 (s) 60.26 (s) 63.45 (s) 72.33 (s) 75.83 (s) 76.93 (s) 80.28 (s) 82.55 (s) 109.62 (s) 125.08 (s) 126.37 (s) 129.18 (s) 130.01 (s) 130.08 (s) 130.70 (s) 131.84 (s) 134.71 (s) 136.28 (s) 137.55 (s) 140.70 (s) 149.76 (s) 158.63 (s) 178.65 (s); HR-MS ESI/APCI Dual m/z: 670.4437 [M + H]⁺ (calcd for C₃₈H₅₉N₃O₇: 670.4426).

(1S)-1,5-Anhydro-1-{5-[(4-{(1E)-4-[(1-{[3-(dimethylamino)propyl]amino}-2-methylpropan-2-yl)amino]-3,3-dimethyl-4-oxobut-1-en-1-yl}-2-methylphenyl)methyl]-2-methoxy-4-(propan-2-yl)phenyl}-D-glucitol (41e)

Compound 36a (350 mg, 0.457 mmol) and N¹,N¹-dimethylpropane-1,3-diamine (56 mg, 0.548 mmol) was used as starting materials and synthesized in the same manner as compound 28a to give the title compound (100 mg, 32% for 2 steps) as a colorless amorphous. ¹H-NMR (600 MHz, CD₃OD) δ ppm 1.14 (d, J = 6.4 Hz, 3H), 1.15 (d, J = 6.4 Hz, 3H), 1.30 (s, 6H), 1.33 (s, 6H), 1.46–1.56 (m, 2H), 2.12 (s, 6H), 2.17–2.24 (m, 2H), 2.33 (s, 3H), 2.51 (t, J = 7.1 Hz, 2H), 2.60 (s, 2H), 2.93–3.03 (m, 1H), 3.37–3.42 (m, 2H), 3.43–3.48 (m, 1H), 3.48–3.55 (m, 1H), 3.57–3.65 (m, 1H), 3.80–3.87 (m, 4H), 3.92 (s, 2H), 4.61 (d, J = 9.2 Hz, 1H), 6.31–6.37 (m, 1H), 6.49 (d, J = 16.5 Hz, 1H), 6.72 (d, J = 7.8 Hz, 1H), 6.92 (s, 1H), 7.07–7.12 (m, 2H), 7.24 (s, 1H); ¹³C-NMR (126 MHz, CD₃OD) δ ppm 20.06 (s) 24.08 (s) 24.24 (s) 25.34 (s) 25.92 (s) 28.49 (s) 30.75 (s) 36.41 (s) 45.57 (s) 46.58 (s) 48.64 (s) 49.76 (s) 54.40 (s) 56.52 (s) 58.72 (s) 60.06 (s) 63.46 (s) 72.34 (s) 75.82 (s) 76.95 (s) 80.30 (s) 82.57 (s) 109.64 (s) 125.03 (s) 126.36 (s) 129.21 (s) 130.07 (s) 130.11 (s) 130.54 (s) 131.83 (s) 134.92 (s) 136.32 (s) 137.55 (s) 140.65 (s) 149.77 (s) 158.64 (s) 178.67 (s); HR-MS ESI/APCI Dual m/z: 684.4611[M + H]⁺ (calcd for C₃₉H₆₁N₃O₇: 684.4582).

(1S)-1,5-Anhydro-1-{5-[(4-{(1E)-4-[(1-{[4-(dimethylamino)butyl]amino}-2-methylpropan-2-yl)amino]-3,3-dimethyl-4-oxobut-1-en-1-yl}-2-methylphenyl)methyl]-2-methoxy-4-(propan-2-yl)phenyl}-D-glucitol (41f)

Compound 36a (500 mg, 0.653 mmol) and N¹,N¹-dimethylbutane-1,4-diamine dihydrochloride (184 mg, 0.979 mmol) was used as starting materials and synthesized in the same manner as compound 28b to give the title compound (66.6 mg, 38% for 2 steps) as a colorless amorphous. ¹H-NMR (600 MHz, CD₃OD) δ ppm 1.11–1.17 (m, 6H), 1.29–1.41 (m, 16H), 2.16–2.23 (m, 8H), 2.33 (s, 3H), 2.51 (t, J = 7.3 Hz, 2H), 2.62 (s, 2H), 2.94–3.02 (m, 1H), 3.33–3.38 (m, 2H), 3.43–3.48 (m, 1H), 3.48–3.54 (m, 1H), 3.61 (dd, J = 12.2, 5.7 Hz, 1H), 3.79–3.87 (m, 4H), 3.92 (s, 2H), 4.61 (d, J = 9.6 Hz, 1H), 6.34 (d, J = 16.0 Hz, 1H), 6.49 (d, J = 16.0 Hz, 1H), 6.74 (d, J = 7.8 Hz, 1H), 6.92 (s, 1H), 7.08 (s, 1H), 7.10 (d, J = 7.8 Hz, 1H), 7.24 (s, 1H); ¹³C-NMR (126 MHz, CD₃OD) δ ppm 20.09 (s) 24.07 (s) 24.22 (s) 25.40 (s) 25.90 (s) 26.08 (s) 28.81 (s) 30.74 (s) 36.45 (s) 45.53 (s) 46.56 (s) 51.36 (s) 54.40 (s) 56.52 (s) 59.91 (s) 60.64 (s) 63.47 (s) 72.34 (s) 75.79 (s) 76.97 (s) 80.28 (s) 82.56 (s) 109.65 (s) 125.05 (s) 126.33 (s) 129.20 (s) 130.07 (s) 130.15 (s) 130.58 (s) 131.76 (s) 134.87 (s) 136.32 (s) 137.51 (s) 140.59 (s) 149.75 (s) 158.63 (s) 178.71 (s); HR-MS ESI/APCI Dual m/z: 698.4783 [M + H]⁺ (calcd for C₄₀H₆₃N₃O₇: 698.4739).

(1S)-1,5-Anhydro-1-{5-[(4-{(1E)-4-[(1-{[4-(dimethylamino)butyl](methyl)amino}-2-methylpropan-2-yl)amino]-3,3-dimethyl-4-oxobut-1-en-1-yl}-2-methylphenyl)methyl]-2-methoxy-4-(propan-2-yl)phenyl}-D-glucitol (41g)

Sodium triacetoxyborohydride (554 mg, 2.61 mmol) and N¹,N¹-dimethylbutane-1,4-diamine dihydrochloride (184 mg, 0.979 mmol) were added to a N,N-dimethylformamide (6 mL) solution of compound 36a (500 mg, 0.653 mmol) in a nitrogen atmosphere, and the mixture was stirred for 16 h at room temperature. An aqueous sodium hydrogen carbonate solution was added to the reaction liquid, and after extracting the mixture with chloroform, the organic layer was washed with 10% brine and then dried over anhydrous magnesium sulfate. The desiccant was filtered out, and the solvent was distilled off under reduced pressure. The resulting residue was purified by performing silica gel column chromatography (chloroform : methanol = 10 : 0 to 9 : 1) to obtain the intermediate (262 mg). To a solution of the intermediate (156 mg), 37% formaldehyde solution (73 mg, 0.90 mmol) and sodium triacetoxyborohydride (153 mg, 0.72 mmol) were added, and stirred for 6 h at room temperature. An aqueous sodium hydrogen carbonate solution was added to the reaction liquid, and after extracting the mixture with chloroform, the organic layer was washed with 10% brine and then dried over anhydrous magnesium sulfate. The desiccant was filtered out, and the solvent was distilled off under reduced pressure. The resulting residue was purified by performing silica gel column chromatography (chloroform : methanol = 10 : 0 to 9 : 1) to obtain the intermediate (141 mg). The crude intermediate (141 mg) in methanol : triethylamine : water (5 : 1 : 1) solution (2.5 mL) was stirred at the room temperature for 16 h, the solvent was distilled off under reduced pressure. The resulting residue was purified by performing neutral silica gel column chromatography (ethyl acetate : ethanol : water = 30 : 2 : 1) to give the title compound (76 mg, 27% in 3steps) as a colorless amorphous. ¹H-NMR (600 MHz, CD₃OD) δ ppm 1.14 (d, J = 6.4 Hz, 6H), 1.25–1.37 (m, 16H), 2.14–2.21 (m, 11H), 2.30–2.36 (m, 5H), 2.41 (s, 2H), 2.93–3.00 (m, 1H), 3.34–3.38 (m, 2H), 3.46 (t, J = 8.7 Hz, 1H), 3.49–3.55 (m, 1H), 3.62 (dd, J = 11.9, 6.0 Hz, 1H), 3.81–3.86 (m, 4H), 3.93 (s, 2H), 4.61 (d, J = 9.6 Hz, 1H), 6.34 (d, J = 16.5 Hz, 1H), 6.51 (d, J = 16.5 Hz, 1H), 6.74 (d, J = 7.8 Hz, 1H), 6.92 (s, 1H), 7.08–7.13 (m, 2H), 7.25 (s, 1H); ¹³C-NMR (126 MHz, CD₃OD) δ ppm 20.12 (s) 24.09 (s) 24.25 (s) 25.91 (s) 26.13 (s) 26.37 (s) 26.79 (s) 30.75 (s) 36.47 (s) 45.19 (s) 45.57 (s) 46.68 (s) 54.50 (s) 56.52 (s) 60.73 (s) 61.17 (s) 63.47 (s) 68.43 (s) 72.34 (s) 75.79 (s) 77.00 (s) 80.29 (s) 82.57 (s) 109.66 (s) 125.05 (s) 126.35 (s) 129.23 (s) 130.06 (s) 130.12 (s) 130.79 (s) 131.86 (s) 134.70 (s) 136.20 (s) 137.52 (s) 140.74 (s) 149.75 (s) 158.65 (s) 178.40 (s); HR-MS ESI/APCI Dual m/z: 712.4916 [M + H]⁺ (calcd for C₄₁H₆₅N₃O₇: 712.4895).

(1S)-1,5-Anhydro-1-[5-({4-[(1E)-4-({1-[4-(2-hydroxyethyl)piperazin-1-yl]-2-methylpropan-2-yl}amino)-3,3-dimethyl-4-oxobut-1-en-1-yl]-2-methylphenyl}methyl)-2-methoxy-4-(propan-2-yl)phenyl]-D-glucitol (41 h)

Compound 36a (300 mg, 0.392 mmol) and 1-piperazine ethanol (76 mg, 0.588 mmol) was used as starting materials and synthesized in the same manner as compound 28b to give the title compound (68.6 mg, 25% for 2 steps) as a colorless amorphous. ¹H-NMR (600 MHz, CD₃OD) δ ppm 1.16 (d, J = 6.9 Hz, 6H), 1.30 (s, 6H), 1.33 (s, 6H), 2.05–2.22 (m, 6H), 2.33 (s, 2H), 2.35–2.38 (m, 3H), 2.47 (br s., 4H), 2.97–3.05 (m, 1H), 3.34–3.39 (m, 2H), 3.43–3.54 (m, 4H), 3.63 (dd, J = 12.2, 5.3 Hz, 1H), 3.80–3.87 (m, 4H), 3.93 (s, 2H), 4.62 (d, J = 9.2 Hz, 1H), 6.38 (d, J = 16.0 Hz, 1H), 6.53 (d, J = 16.0 Hz, 1H), 6.75 (d, J = 7.8 Hz, 1H), 6.93 (s, 1H), 7.11 (s, 1H), 7.15 (d, J = 7.8 Hz, 1H), 7.28 (s, 1H); ¹³C-NMR (126 MHz, CD₃OD) δ ppm 20.12 (s) 24.08 (s) 24.32 (s) 26.02 (dd) 30.77 (s) 36.28 (s) 46.72 (s) 54.58 (s) 54.71 (s) 55.93 (s) 56.50 (s) 59.75 (s) 61.21 (s) 63.34 (s) 68.64 (s) 72.23 (s) 75.75 (s) 76.95 (s) 80.30 (s) 82.62 (s) 109.60 (s) 125.15 (s) 126.32 (s) 129.49 (s) 130.01 (s) 130.24 (s) 130.87 (s) 131.83 (s) 134.41 (s) 136.07 (s) 137.72 (s) 141.03 (s) 149.79 (s) 158.64 (s) 178.47 (s); HR-MS ESI/APCI Dual m/z: 712.4561 [M + H]⁺ (calcd for C₄₀H₆₁N₃O₈: 712.4531).

(1S)-1,5-Anhydro-1-[5-({4-[(1E)-4-{[1-(4-carbamoylpiperidin-1-yl)-2-methylpropan-2-yl]amino}-3,3-dimethyl-4-oxobut-1-en-1-yl]-2-methylphenyl}methyl)-2-methoxy-4-(propan-2-yl)phenyl]-D-glucitol (41i)

Compound 36a (150 mg, 0.196 mmol) and piperidine-4-carboxamide (38 mg, 0.294 mmol) was used as starting materials and synthesized in the same manner as compound 28b to give the title compound (32 mg, 23% for 2 steps) as a colorless amorphous. ¹H-NMR (300 MHz, CD₃OD) δ ppm 1.09–1.21 (m, 6H), 1.29 (s, 6H), 1.34 (s, 6H), 1.43–1.59 (m, 4H), 1.98–2.13 (m, 1H), 2.15–2.28 (m, 2H), 2.33 (s, 3H), 2.41 (s, 2H), 2.73–2.86 (m, 2H), 2.89–3.02 (m, 1H), 3.33–3.67 (m, 5H), 3.78–3.86 (m, 4H), 3.91 (s, 2H), 4.62 (d, J = 9.5 Hz, 1H), 6.35 (d, J = 16.2 Hz, 1H), 6.50 (d, J = 16.2 Hz, 1H), 6.67–6.76 (m, 1H), 6.92 (s, 1H), 7.08–7.16 (m, 2H), 7.25 (s, 1H); ¹³C-NMR (126 MHz, CD₃OD) δ ppm 20.01 (s) 24.09 (s) 24.21 (s) 25.81 (s) 25.84 (s) 25.87 (s) 30.28 (s) 30.73 (s) 36.40 (s) 43.09 (s) 46.63 (s) 55.04 (s) 56.52 (s) 56.70 (s) 56.72 (s) 63.39 (s) 68.25 (s) 72.30 (s) 75.65 (s) 76.96 (s) 80.29 (s) 82.57 (s) 109.66 (s) 125.03 (s) 126.21 (s) 129.22 (s) 130.13 (s) 130.16 (s) 130.77 (s) 131.87 (s) 134.52 (s) 136.01 (s) 137.65 (s) 140.79 (s) 149.84 (s) 158.63 (s) 178.66 (s) 180.83 (s); HR-MS ESI/APCI Dual m/z: 710.4399 [M + H]⁺ (calcd for C₄₀H₅₉N₃O₈: 710.4375).

(1S)-1,5-Anhydro-1-[5-({4-[(1E)-4-({1-[4-(dimethylamino)piperidin-1-yl]-2-methylpropan-2-yl}amino)-3,3-dimethyl-4-oxobut-1-en-1-yl]-2-methylphenyl}methyl)-2-methoxy-4-(propan-2-yl)phenyl]-D-glucitol (41j)

Sodium triacetoxyborohydride (166 mg, 0.784 mmol) and N,N-dimethylpiperidin-4-amine (37.7 mg, 0.294 mmol) were added to a chloroform (2 mL) solution of compound 36a (150 mg, 0.196 mmol) in a nitrogen atmosphere, and the mixture was stirred for 16 h at room temperature. An aqueous sodium hydrogen carbonate solution was added to the reaction liquid, and after extracting the mixture with chloroform, the organic layer was washed with 10% brine and then dried over anhydrous magnesium sulfate. The desiccant was filtered out, and the solvent was distilled off under reduced pressure. The resulting residue was purified by performing silica gel column chromatography (chloroform : methanol = 10 : 0 to 9 : 1) to obtain the intermediate (97.6 mg). The crude intermediate (97 mg) in methanol : triethylamine : water (5 : 1 : 1) solution (2.5 mL) was stirred at the room temperature for 16 h, and after stirring the reaction mixture for 16 h at room temperature, the solvent was distilled off under reduced pressure. The resulting residue was purified by performing neutral silica gel column chromatography (ethyl acetate : ethanol : water = 30 : 2 : 1) to give the title compound (56 mg, 41% for 2 steps) as a colorless amorphous. ¹H-NMR (600 MHz, CD₃OD) δ ppm 1.13 (d, J = 6.4 Hz, 3H), 1.14 (d, J = 6.4 Hz, 3H), 1.19–1.27 (m, 2H), 1.29 (s, 6H), 1.33 (s, 6H), 1.47–1.54 (m, 2H), 1.98–2.05 (m, 1H), 2.14 (s, 6H), 2.34 (m, 5H), 2.76–2.82 (m, 2H), 2.96 (t, J = 6.7 Hz, 1H), 3.35–3.38 (m, 2H), 3.44–3.48 (m, 1H), 3.48–3.54 (m, 1H), 3.62 (dd, J = 11.9, 6.0 Hz, 1H), 3.80–3.87 (m, 4H), 3.91 (s, 2H), 4.62 (d, J = 9.6 Hz, 1H), 6.37 (d, J = 16.5 Hz, 1H), 6.53 (d, J = 16.5 Hz, 1H), 6.72 (d, J = 7.8 Hz, 1H), 6.93 (s, 1H), 7.09 (s, 1H), 7.14 (d, J = 7.8 Hz, 1H), 7.26 (s, 1H); ¹³C-NMR (126 MHz, CD₃OD) δ ppm 20.08 (s) 24.10 (s) 24.24 (s) 25.79 (s) 25.85 (s) 29.55 (s) 29.65 (s) 30.77 (s) 36.41 (s) 41.68 (s) 46.68 (s) 55.06 (s) 56.52 (s) 56.65 (s) 63.25 (s) 63.47 (s) 68.00 (s) 72.35 (s) 75.83 (s) 76.93 (s) 80.29 (s) 82.57 (s) 109.64 (s) 124.90 (s) 126.39 (s) 130.02 (dd) 131.88 (s) 134.64 (s) 136.13 (s) 137.61 (s) 140.82 (s) 149.76 (s) 158.65 (s) 178.40 (s); HR-MS ESI/APCI Dual m/z: 710.4767 [M + H]⁺ (calcd for C₄₁H₆₃N₃O₇: 710.4739). Purity and retention time of 41j were determined using LC-MS (Agilent 1290 Infinity) under the following conditions. Column: Acquity CSH C18, 1.7 µm, 2.1 × 50 mm of Waters. Ionization method: ESI. Solvents: Fluid A, 0.1% formic acid containing water; Fluid B, 0.1% formic acid containing acetonitrile. Flow rate: 0.8 mL/min. Detection method: 254 nm. Gradient: 0.0–0.8 min (Fluid A / Fluid B = 95/5–60/40), 0.8–1.08 min (Fluid A / Fluid B = 60/40–1/99), 1.08–1.38 min (Fluid A / Fluid B = 1/99). Retention Time = 0.952 min; Purity = 95.8%.

Acknowledgments

The authors thank Katsumasa Otake for helpful suggestions and collection of the ADME data.

Conflict of Interest

The authors declare no conflict of interest.

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