NOVEL TRIAZINE-BASED AMPHIPHILIC COMPOUND AND USE THEREOF

Abstract

The present invention relates to: a newly developed triazine-cored amphiphilic compound; a preparation method therefor; and a method for extracting, solubilizing, stabilizing, crystallizing or analyzing membrane proteins by using the same. In addition, the compound enables membrane proteins, which have various structures and characteristics, to be efficiently extracted from cell membranes and stably stored in an aqueous solution for a long time, compared to a conventional compound, thereby being usable in functional and structural analysis thereof. Analyzing the structure and function of membrane proteins is closely related to the development of a novel drug, and thus is one of the greatest interests in the biology and chemistry fields.

Claims

1. A compound represented by the following Chemical Formula 1 or an isomer thereof: ##STR00006## in Chemical Formula 1, R.sup.1 and R.sup.2 are each independently a substituted or unsubstituted C.sub.3-C.sub.30 alkyl group; E.sup.1 and E.sup.2 are each independently NH, O or S; Z is a direct bond or NH; Y is CH or N; L.sup.1 and L.sup.2 are each independently a C.sub.1-5 alkylene group; X.sup.1 and X.sup.2 are each independently a saccharide linked to oxygen; and when Y is CH, Y is optionally further unsubstituted or substituted with a substituent represented by -L.sup.3X.sup.3, where L.sup.3 is a C.sub.1-5 alkylene group, and X.sup.3 is a saccharide linked to oxygen.

2. The compound or the isomer thereof of claim 1, wherein the saccharide is a monosaccharide or a disaccharide.

3. The compound or the isomer thereof of claim 1, wherein the saccharide is glucose or maltose.

4. The compound or the isomer thereof of claim 1, wherein R.sup.1 and R.sup.2 are each independently a substituted or unsubstituted C.sub.3-C.sub.20 alkyl group; E.sup.1 and E.sup.2 are each independently NH, O or S; L.sup.1 and L.sup.2 are each independently a C.sub.1-3 alkylene group; X.sup.1 and X.sup.2 are maltose; Y is CH; and Z is NH.

5. The compound or the isomer thereof of claim 1, wherein R.sup.1 and R.sup.2 are each independently a substituted or unsubstituted C.sub.3-C.sub.20 alkyl group; E.sup.1 and E.sup.2 are each independently NH, O or S; L.sup.1 and L.sup.2 are each independently a C.sub.1-3 alkylene group; X.sup.1 and X.sup.2 are glucose; Z is NH; and Y is CH, where Y is further substituted with a substituent represented by -L.sup.3X.sup.3, L.sup.3 is a C.sub.1-3 alkylene group, and X.sup.3 is glucose.

6. The compound or the isomer thereof of claim 1, wherein R.sup.1 and R.sup.2 are each independently a substituted or unsubstituted C.sub.3-C.sub.20 alkyl group; E.sup.1 and E.sup.2 are each independently NH, O or S; L.sup.1 and L.sup.2 are each independently a C.sub.1-3 alkylene group; X.sup.1 and X.sup.2 are maltose; Y is N; and Z is a direct bond.

7. The compound or the isomer thereof of claim 1, wherein the compound is represented by one of the following Chemical Formulae 2 to 8: ##STR00007## ##STR00008## in the formulae, R.sup.1 and R.sup.2 are each independently a substituted or unsubstituted C.sub.5-C.sub.20 alkyl group.

8. The compound or the isomer thereof of claim 1, wherein the compound is an amphiphilic molecule for extracting, solubilizing, stabilizing, crystallizing or analyzing a membrane protein.

9. The compound or the isomer thereof of claim 1, wherein the compound has a critical micelle concentration (CMC) of 0.0001 to 1 mM in an aqueous solution.

10. A composition for extracting, solubilizing, stabilizing, crystallizing or analyzing a membrane protein, comprising the compound or the isomer thereof of claim 1.

11. The composition of claim 10, wherein the composition is a formulation of micelles, liposomes, emulsions or nanoparticles.

12. A method for preparing a compound represented by the following Chemical Formula 1, the method comprising: 1) introducing an alkyl group by reacting an alkylamine, alcohol or thiol with 2,4,6-trichloro-1,3,5-triazine; 2) introducing a hydroxyl end by reacting an amine substituted with at least two hydroxyalkyls or an alkylamine substituted with at least two hydroxyalkyls with the product of step 1); 3) introducing a saccharide to which a protecting group is attached by performing a glycosylation reaction on the product of step 2); and 4) performing a deprotection reaction on the product of step 3): ##STR00009## in Chemical Formula 1, R.sup.1 and R.sup.2 are each independently a substituted or unsubstituted C.sub.3-C.sub.30 alkyl group; E.sup.1 and E.sup.2 are each independently NH, O or S; Z is a direct bond or NH; Y is CH or N; L.sup.1 and L.sup.2 are each independently a C.sub.1-5 alkylene group; X.sup.1 and X.sup.2 are each independently a saccharide linked to oxygen; and when Y is CH, Y is optionally further unsubstituted or substituted with a substituent represented by -L.sup.3X.sup.3, where L.sup.3 is a C.sub.1-5 alkylene group, and X.sup.3 is a saccharide linked to oxygen.

13. The method of claim 12, wherein R.sup.1 and R.sup.2 are each independently a substituted or unsubstituted C.sub.3-C.sub.20 alkyl group; E.sup.1 and E.sup.2 are each independently NH, O or S; L.sup.1 and L.sup.2 are each independently a C.sub.1-3 alkylene group; X.sup.1 and X.sup.2 are maltose; Y is CH; and Z is NH.

14. The method of claim 12, wherein R.sup.1 and R.sup.2 are each independently a substituted or unsubstituted C.sub.3-C.sub.20 alkyl group; E.sup.1 and E.sup.2 are each independently NH, O or S; L.sup.1 and L.sup.2 are each independently a C.sub.1-3 alkylene group; X.sup.1 and X.sup.2 are glucose; Z is NH; and Y is CH, where Y is further substituted with a substituent represented by -L.sup.3X.sup.3, L.sup.3 is a C.sub.1-3 alkylene group, and X.sup.3 is glucose.

15. The method of claim 12, wherein R.sup.1 and R.sup.2 are each independently a substituted or unsubstituted C.sub.3-C.sub.20 alkyl group; E.sup.1 and E.sup.2 are each independently NH, O or S; L.sup.1 and L.sup.2 are each independently a C.sub.1-3 alkylene group; X.sup.1 and X.sup.2 are maltose; Y is N; and Z is a direct bond.

16. A method for extracting, solubilizing, stabilizing, crystallizing or analyzing a membrane protein, the method comprising: treating the membrane protein with a compound represented by the following Chemical Formula 1 or an isomer thereof in an aqueous solution: ##STR00010## in Chemical Formula 1, R.sup.1 and R.sup.2 are each independently a substituted or unsubstituted C.sub.3-C.sub.30 alkyl group; E.sup.1 and E.sup.2 are each independently NH, O or S; Z is a direct bond or NH; Y is CH or N; L.sup.1 and L.sup.2 are each independently a C.sub.1-5 alkylene group; X.sup.1 and X.sup.2 are each independently a saccharide linked to oxygen; and when Y is CH, Y is optionally further unsubstituted or substituted with a substituent represented by -L.sup.3X.sup.3, where L.sup.3 is a C.sub.1-5 alkylene group, and X.sup.3 is a saccharide linked to oxygen.

17. The method of claim 16, wherein R.sup.1 and R.sup.2 are each independently a substituted or unsubstituted C.sub.3-C.sub.20 alkyl group; E.sup.1 and E.sup.2 are each independently NH, O or S; L.sup.1 and L.sup.2 are each independently a C.sub.1-3 alkylene group; X.sup.1 and X.sup.2 are maltose; Y is CH; and Z is NH.

18. The method of claim 16, wherein R.sup.1 and R.sup.2 are each independently a substituted or unsubstituted C.sub.3-C.sub.20 alkyl group; E.sup.1 and E.sup.2 are each independently NH, O or S; L.sup.1 and L.sup.2 are each independently a C.sub.1-3 alkylene group; X.sup.1 and X.sup.2 are glucose; Z is NH; and Y is CH, where Y is further substituted with a substituent represented by -L.sup.3X.sup.3, L.sup.3 is a C.sub.1-3 alkylene group, and X.sup.3 is glucose.

19. The method of claim 16, wherein R.sup.1 and R.sup.2 are each independently a substituted or unsubstituted C.sub.3-C.sub.20 alkyl group; E.sup.1 and E.sup.2 are each independently NH, O or S; L.sup.1 and L.sup.2 are each independently a C.sub.1-3 alkylene group; X.sup.1 and X.sup.2 are maltose; Y is N; and Z is a direct bond.

20. The method of claim 16, wherein the membrane protein is a leucine transporter (LeuT), a human ?.sub.2 adrenergic receptor (?.sub.2AR), a melibiose permease (MelB), boron transporter 1 (BOR1), a mouse u-opioid receptor (MOR) or a combination of two or more thereof.

Description

DESCRIPTION OF DRAWINGS

[0089] FIG. 1 is a view illustrating a synthesis scheme of TSMs according to Example 1 of the present invention.

[0090] FIG. 2 is a view illustrating a synthesis scheme of TTGs according to Example 2 of the present invention.

[0091] FIG. 3 is a view illustrating a synthesis scheme of TEMs according to Example 3 of the present invention.

[0092] FIG. 4 is a set of views illustrating the size distribution map of micelles formed by TSMs and TEMs.

[0093] FIG. 5 is a set of views illustrating the size distribution map of micelles formed by TSMs and TEMs.

[0094] FIG. 6 illustrates the results of measuring leucine transporter (LeuT) structural stability in an aqueous solution by TSMs, TEMs or DDM at CMC+0.04 wt %. Protein stability was confirmed by measuring the substrate binding properties of the transporter via scintillation proximity assay (SPA). The substrate binding properties of the proteins were measured at regular intervals while incubating LeuT for 13 days at room temperature in the presence of each amphiphilic compound.

[0095] FIG. 7 illustrates the results of measuring leucine transporter (LeuT) structural stability in an aqueous solution by TSMs, TEMs or DDM at CMC+0.2 20 wt %. Protein stability was confirmed by measuring the substrate binding properties of the transporter via scintillation proximity assay (SPA). The substrate binding properties of the proteins were measured at regular intervals while incubating LeuT for 12 days at room temperature in the presence of each amphiphilic compound.

[0096] FIG. 8 illustrates the results of measuring leucine transporter structural stability in an aqueous solution by TTGs or DDM at (A) CMC+0.04 wt % or (B) CMC+0.2 wt %. Protein stability was confirmed by measuring the substrate binding properties of the transporter via scintillation proximity assay (SPA). The substrate binding properties of the proteins were measured at regular intervals while incubating LeuT for 12 days at room temperature in the presence of each amphiphilic compound.

[0097] FIG. 9 illustrates the results of, after extracting MelB protein at 0? C. for 90 minutes using TEMs or DDM, further incubating the extracted protein at four high temperatures (0, 45, 55, and 65? C.) for 90 minutes, and then measuring the amount of MelB protein dissolved in an aqueous solution: [0098] (A) SDS-PAGE and Western Blotting results showing the amount of MelB protein extracted using each amphiphilic compound; and [0099] (B) A histogram of showing the amount of MelB protein extracted using each amphiphilic compound as a percentage (%) of the total amount of protein present in a membrane sample (Memb) untreated with an amphiphilic compound.

[0100] FIG. 10 illustrates the results of, after extracting MelB protein at 0? C. for 90 minutes using TSMs or DDM, further incubating the extracted protein at four high temperatures (0, 45, 55, and 65? C.) for 90 minutes, and then measuring the amount of MelB protein dissolved in an aqueous solution: [0101] (a) SDS-PAGE and Western Blotting results showing the amount of MelB protein extracted using each amphiphilic compound; and [0102] (B) A histogram of showing the amount of MelB protein extracted using each amphiphilic compound as a percentage (%) of the total amount of protein present in a membrane sample (Memb) untreated with an amphiphilic compound.

[0103] FIG. 11 illustrates the results of, after extracting MelB protein at 0? C. for 90 minutes using TTGs or DDM, further incubating the extracted protein at four high temperatures (0, 45, 55, and 65? C.) for 90 minutes, and then measuring the amount of MelB protein dissolved in an aqueous solution: [0104] (A) SDS-PAGE and Western Blotting results showing the amount of MelB protein extracted using each amphiphilic compound; and [0105] (B) A histogram of showing the amount of MelB protein extracted using each amphiphilic compound as a percentage (%) of the total amount of protein present in a membrane sample (Memb) untreated with an amphiphilic compound.

[0106] FIG. 12 illustrates the results of measuring the initial (30 minutes after amphiphilic molecular exchange) effect of CMCs 30 0.2 wt % of TSMs, TEMs or DDM on the stability of ?.sub.2AR. The ligand binding properties of the receptor were measured via ligand binding assay of [.sup.3H]-dihydroalprenolol (DHA).

[0107] FIG. 13 illustrates the results of measuring the long-term effect of CMCs+0.2 wt % of TSMs, TEMs or DDM on the stability of ?.sub.2AR over the passage of measurement time (1 h, 8 h, 1 day, 2 days, 3 days, and 6 days). The ligand binding properties of the receptor were measured via ligand binding assay of [.sup.3H]-dihydroalprenolol (DHA), and measured by sampling a protein sample at regular intervals while storing the protein sample at room temperature for 6 days.

[0108] FIG. 14 illustrates the results of measuring the initial (30 minutes after amphiphilic molecular exchange) effect (A) and the long-term effect of CMCs+0.2 wt % of TTGs or DDM on the stability of ?.sub.2AR over the passage of measurement time (1 h, 8 h, 1 day, 2 days, 3 days, and 6 days) (B). The ligand binding properties of the receptor were measured via ligand binding assay of [.sup.3H]-dihydroalprenolol (DHA).

[0109] FIG. 15 illustrates a set of thermal denaturation profiles of AtBOR1 solubilized by CMCs+0.04 wt % of TSMs (TSM-E9/E10/T8/T9), TEMs (TEM-E9/E10/T8/T9) or DDM over the passage of time (A, B). These results were measured via CPM performed at 40? C. for 120 minutes. Further, FIG. 15 illustrates a set of results of testing the thermal stability of a protein extracted by the amphiphilic molecule using fluorescence size exclusion chromatography (FSEC)(C, 5 D).

[0110] FIG. 16 illustrates a set of thermal denaturation profiles of AtBOR1 solubilized by CMCs++0.2 wt % of TSMs (TSM-E9/E10/T8/T9), TEMs (TEM-E9/E10/T8/T9) or DDM over the passage of time.

[0111] FIG. 17 illustrates the thermal stability profile (A) and melting point (Tm) (B) of MOR dissolved in TSMs (TSM-E9/E10/T8/T9), TEMs (TEM-E9/E10/T8/T9) or DDM.

MODES OF THE INVENTION

[0112] Hereinafter, the present invention will be described in more detail in the following Examples. However, the following Examples merely exemplify the content of the present invention, and do not limit or restrict the scope of rights of the present invention. From the detailed description and examples of the present invention, it is understood that what can be easily inferred by a person skilled in the art to which the present invention pertains belongs to the scope of rights of the present invention.

EXAMPLE 1

Synthesis Method of TSMs

[0113] FIG. 1 illustrates the synthesis scheme of TSMs. 10 types of compounds of resorcinarene-based maltosides (TSMs) were synthesized by the following synthesis methods of <1-1>to <1-4>.

<1-1> General Procedure for the Synthesis of 2-chloro-4,6-dialkylated-1,3,5-triazine (Step i of FIG. 1)

[0114] A mixture of 2,4,6-trichloro-1,3,5-triazine (3.01 mmol) and NaHCO.sub.3 (7.26 mmol) was stirred in acetone (10 mL) for 10 minutes. Each alcohol (ROH/RSH) (6.0 mmol) dissolved in acetone was added dropwise for 30 minutes. The resulting reaction mixture was kept at room temperature for 36 hr for ROH or 1 hr for RSH. The reaction mixture was extracted with CHCl.sub.3 and water, and the organic layer was dried over anhydrous Na.sub.2SO.sub.4. The oily residue obtained after removal of solvent was subjected to column chromatography purification to obtain target Compound 1 or 3.

<1-2> General Synthesis Procedure for Coupling Reactions of the Resulting Dialkylated Triazine Derivatives With 2-amino-1,3-propanediol (Step ii of FIG. 1)

[0115] To a mixture of 2-chloro-4,6-dialkylated-1,3,5-triazine (1.0 equiv.) dissolved in THF, 2-amino-1,3-propanediol and K.sub.2CO.sub.3 were added under nitrogen. The solution was stirred at 40? C. for 24 hr. The reaction mixture was diluted with water and then extracted with ethyl acetate. The organic layer was washed with brine and dried over anhydrous Na.sub.2SO.sub.4. After evaporation of the ethyl acetate solution, the residue was purified by flash column chromatography (EtOAc/hexane) to obtain target Compound 2 or 4.

<1-3> General Synthesis Procedure for Glycosylation Reaction (Step iii of FIG. 1)

[0116] This reaction was performed according to the synthesis method (Nat. Methods 2010, 7, 1003.) of Chae, P. S. et al. with some modifications. Briefly, a mixture of a dialkylated di-ol derivative (Compound 2 or 4), AgOTf (2.5 equiv.), 2,4,6-collidine (0.5 equiv.) in anhydrous CH.sub.2Cl.sub.2 (20 mL) was stirred at ?45? C. Then, perbenzoylated maltosylbromide (2.5 equiv.) dissolved in CH.sub.2Cl.sub.2 (30 mL) was added dropwise over 0.5 hr into this suspension. The reaction was maintained at 0? C. for 1.5 hr. Reaction progress was monitored by TLC. After completion of the reaction (as detected by TLC), pyridine (1.0 mL) was added to the reaction mixture. The reaction mixture was diluted with CH.sub.2Cl.sub.2 (30 mL) before being filtered over Celite. The filtrate was washed successively with a 1.0 M aqueous Na.sub.2S.sub.2O.sub.3 solution (30 mL), a 0.1 M aqueous HCl solution (30 mL), and brine (30 mL). Then, the organic layer was dried with anhydrous Na.sub.2SO.sub.4 and the solvent was removed by rotary evaporation. The resulting residue was purified by silica gel column chromatography (EtOAc/hexane) to obtain the glycosylated target compound.

<1-4> General Synthesis Procedure for Deprotection Reaction (Step iv of FIG. 1)

[0117] This reaction followed the synthesis method (Nat. Methods 2010, 7, 1003.) of Chae, P. S. aureus et al. The de-O-benzoylation was performed under Zemplen's condition. An 0-protected compound was dissolved in anhydrous CH.sub.2Cl.sub.2 and then MeOH was added slowly thereto until persistent precipitation appeared. A methanolic solution of 0.5 M NaOMe was added to the reaction mixture such that the final concentration of NaOMe was 0.05 M. The reaction mixture was stirred for 6 hr at room temperature. After completion of the reaction, the reaction mixture was neutralized using Amberlite IR-120 (H.sup.+ form) resin. The resin was removed by filtration and washed with MeOH and the solvent was removed from the filtrate in vacuo. The residue was purified by silica gel column chromatography (CH.sub.2Cl.sub.2/MeOH) to obtain the target compound.

Preparation Example 1

Synthesis of TSM-E7

<1-1> Synthesis of Compound 1a

[0118] Compound 1a was synthesized in 48% yield according to Example 1-1. .sup.1H NMR (400 MHz, CDCl.sub.3): ? 4.41 (t, J=6.6 Hz, 4H), 1.82-1.75 (m, 4H), 1.34-1.27 (m, 16H), 0.88 (t, J=6.9 Hz, 6H); .sup.13C NMR (100 MHz, CDCl.sub.3): ? 172.7, 172.2, 69.6, 32.0, 29.6, 28.6, 25.9, 22.8, 14.2.

<1-2> Synthesis of Compound 2a

[0119] Compound 2a was synthesized in 90% yield according to Example 1-2. .sup.1H NMR (400 MHz, CDCl.sub.3): ? 6.41 (d, J=8.0 Hz, 1H), 4.30-4.24 (m, 4H), 4.15-4.12 (m, 4H), 3.90-3.81 (m, 4H), 1.74-1.72 (m, 4H), 1.39-1.27 (m, 16H), 0.87 (t, J=7.0 Hz, 6H); .sup.13C NMR (100 MHz, CDCl.sub.3): ? 172.1, 171.6, 167.9, 68.0, 67.8, 62.8, 53.2, 31.9, 29.1, 28.8, 25.9, 22.7, 14.2.

<1-3>Synthesis of TSM-E7a

[0120] TSM-E7a was synthesized in 85% yield according to the general glycosylation reaction procedure of Example 1-3. .sup.1H NMR (400 MHz, CDCl.sub.3): ? 8.07-7.95 (m, 16H), 7.89-7.87 (d, J=7.5 Hz, 4H), 7.82-7.80 (d, J=8.2 Hz, 4H), 7.77-7.74 (m, 4H), 7.63-7.21 (m, 42H), 6.12 (t, J=10.0 Hz, 2H), 5.71-5.66 (m, 4H), 5.49 (d, J=9.7 Hz, 1H), 5.35-5.30 (m, 2H), 5.17-5.08 (m, 4H), 4.47-4.53 (m, 4H), 4.34-4.10 (m, 12H), 3.76 (d, J=8.3 Hz, 2H), 3.24-3.19 (m, 2H), 2.97 (t, J=9.3 Hz, 2H), 2.80 (t, J=8.4 Hz, 1H), 1.71-1.64 (m, 4H), 1.39-1.23 (m, 16H), 0.88 (t, J=7.2 Hz, 3H), 0.87 (t, J=7.2 Hz, 3H); .sup.13C NMR (100 MHz, CDCl.sub.3): ? 172.2, 171.9, 167.5, 166.2, 165.9, 165.8, 165.5, 165.1, 164.9, 134.0, 133.7, 133.5, 133.4, 133.2, 130.0, 129.9, 129.8, 129.7, 129.7, 129.5, 129.4, 129.3, 129.0, 128.9, 128.8, 128.7,128.5, 128.4, 71.0, 128.3, 100.9, 95.6, 77.5, 76.9, 71.8, 71.3, 69.8, 69.1, 69.0, 67.6, 62.6, 31.8, 31.7, 29.0, 28.8, 28.7, 25.9, 22.7, 21. 114.2.

<1-4>Synthesis of TSM-E7

[0121] TSM-E7 was synthesized in 90% yield according to the general synthesis procedure for the deprotection reaction of Example 1-4. .sup.1H NMR (400 MHz, CD.sub.3OD): ? 5.17 (d, J=7.0 Hz, 2H), 4.40 (d, J=8.0 Hz, 2H), 4.30-4.16 (m, 4H), 3.97-3.93 (m, 1H), 3.86-3.85 (d, J=7.0 Hz, 2H), 3.86-3.84-3.77 (m, 2H), 3.72-3.69 (m, 4H), 3.59-3.40 (m, 10H), 3.36-3.28 (m, 4H), 3.19-3.14 (m, 4H), 1.65-1.59 (m, 4H), 1.32-1.19 (m, 16H), 0.78 (t, J=7.2 Hz, 6H); .sup.13C NMR (100 MHz, CD.sub.3OD): ? 173.2, 172.81, 169.0, 104.9, 104.7, 103.0, 81.3, 81.2, 77.7, 76.7, 75.1, 74.8, 74.7, 74.2, 71.5, 68.3, 68.9, 68.7, 62.8, 62.2, 32.0, 33.0, 30.7, 30.5, 30.2, 30.0, 27.1, 27.0, 23.9, 23.7, 14.6, 14.5; HRMS (EI): For C.sub.44H.sub.78N.sub.4O.sub.24 [M+Na].sup.+ 1069.4904, found 1069.4908.

Preparation Example 2

Synthesis of TSM-E8

<2-1> Synthesis of Compound 1b

[0122] Compound 1b was synthesized in 45% yield according to Example 1-1. .sup.1H NMR (400 MHz, CDCl.sub.3): ? 4.41 (t, J=6.6 Hz, 4H), 1.81-1.72 (m, 4H), 1.34-1.27 (m, 20H), 0.86 (t, J=6.9 Hz, 6H); .sup.13C NMR (100 MHz, CDCl.sub.3): ? 172.7, 172.2, 69.6, 32.0, 29.7, 29.6, 28.6, 25.9, 22.8, 14.2.

<2-2>Synthesis of Compound 2b

[0123] Compound 2b was synthesized in 89% yield according to Example 1-2. .sup.1H NMR (400 MHz, CDCl.sub.3): ? 6.41 (d, J=8.0 Hz, 1H), 4.30-4.24 (m, 4H), 4.15-4.12 (m, 4H), 3.90-3.81 (m, 4H), 1.74-1.72 (m, 4H), 1.39-1.27 (m, 20H), 0.87 (t, J=7.0 Hz, 6H); .sup.13C NMR (100 MHz, CDCl.sub.3): ? 172.1, 171.6, 167.9, 68.0, 67.8, 62.8, 53.2, 31.9, 29.1, 28.8, 25.9, 22.7, 14.2.

<2-3> Synthesis of TSM-E8a

[0124] TSM-E8a was synthesized in 83% yield according to the general glycosylation reaction procedure of Example 1-3. .sup.1H NMR (400 MHz, CDCl.sub.3): ? 8.07-7.95 (m, 16H), 7.89-7.87 (d, J=7.5 Hz, 4H), 7.82-7.80 (d, J=8.2 Hz, 4H), 7.77-7.74 (m, 4H), 7.63-7.21 (m, 42H), 6.08 (t, J=9.2 Hz 2H), 5.77-5.65 (m, 4H), 5.49 (d, J=9.7 Hz, 1H), 5.36-5.30 (m, 2H), 5.18-5.08 (m, 4H), 4.47-4.53 (m, 4H), 4.35-4.10 (m, 12H), 3.76 (d, J=8.3, Hz 2H), 3.24-3.19 (m, 2H), 2.97 (t, J=7.3 Hz, 2H), 2.80 (t, J=8.4 Hz, 1H), 1.71-1.64 (m, 4H), 1.39-1.23 (m, 20H), 0.88 (t, J=7.2 Hz, 3H), 0.87 (t, J=7.2 Hz, 3H); .sup.13C NMR (100 MHz, CDCl.sub.3): ? 172.2, 171.9, 167.5, 166.2, 165.9, 165.8, 165.5, 165.1, 164.9, 134.0, 133.7, 133.5, 133.4, 133.2, 130.0, 129.9, 129.8, 129.7, 129.7, 129.5, 129.4, 129.3, 129.0, 128.9, 128.8, 128.7, 128.5, 128.4, 71.0, 128.3, 100.9, 95.6, 77.5, 76.9, 71.8, 71.3, 69.8, 69.1, 69.0, 67.6, 62.6, 31.8, 31.7, 29.0, 28.8, 28.7, 25.9, 22.7, 21.1, 14.2.

<2-4> Synthesis of TSM-E8

[0125] TSM-E8 was synthesized in 90% yield according to the general synthesis procedure for the deprotection reaction of Example 1-4. .sup.1H l NMR (400 MHz, CD.sub.3OD): ? 5.17 (d, J=7.0 Hz, 2H), 4.37 (d, J=8.0 Hz, 2H), 4.32-4.21 (m, 4H), 4.00-3.96 (m, 1H), 3.89-3.88 (m, 2H), 3.83-3.79 (m, 2H), 3.75-3.70 (m, 4H), 3.61-3.42 (m, 10H), 3.37-3.30 (m, 4H), 3.20-3.16 (m, 4H), 1.68-1.62 (m, 4H), 1.36-1.22 (m, 20H), 0.88 (t, J=7.2 Hz, 3H), 0.87 (t, J=7.2 Hz, 3H); .sup.13C NMR (100 MHz, CD.sub.3OD): ? 173.2, 172.8, 169.0, 104.9, 104.7, 103.0, 81.3, 81.2, 77.7, 76.7, 75.1, 74.8, 74.7, 74.2, 71.5, 68.3, 68.9, 68.7, 62.8, 62.2, 32.0, 33.0, 33.1, 30.7, 30.5, 30.2, 30.0, 29.9, 27.1, 27.1,27.0, 23.9, 23.7, 14.6, 14.5; HRMS (FAF.sup.30): For C.sub.46H.sub.82N.sub.4O.sub.24 [M+Na].sup.+ 1097.5217, found 1097.5220.

<Preparation Example 3

Synthesis of TSM-E9

<3-1> Synthesis of Compound 1c

[0126] Compound 1c was synthesized in 50% yield according to Example 1-1. .sup.1H NMR (400 MHz, CDCl.sub.3): ? 4.41 (t, J=6.6 Hz, 4H), 1.82-1.72 (m, 4H), 1.24-1.27 (m, 24H), 0.88 (t, J=6.9 Hz, 6H); .sup.13C NMR (100 MHz, CDCl.sub.3): ? 172.7, 172.2, 69.6, 32.0, 29.6, 29.4, 28.6, 25.9, 22.8, 14.2.

<3-2> Synthesis of Compound 2c

[0127] Compound 2c was synthesized in 90% yield according to Example 1-2. .sup.1H NMR (400 MHz, CDCl.sub.3): ? 6.53 (d, J=8.0 Hz, 1H), 4.60-4.24 (m, 4H), 4.16-4.14 (m, 1H), 3.86-3.81 (m, 4H), 1.73-1.71 (m, 4H), 1.38-1.26 (m, 24H), 0.87 (t, J=6.8 Hz, 6H); .sup.13C NMR (100 MHz, CDCl.sub.3): ? 172.0, 171.4, 167.7, 67.9, 67.7, 62.1, 53.3, 31.9, 29.6, 29.4, 28.8, 25.9, 22.7, 14.1.

<3-3> Synthesis of TSM-E9a

[0128] TSM-E9a was synthesized in 85% yield according to the general glycosylation reaction procedure of Example 1-3. .sup.1H NMR (400 MHz, CDCl.sub.3): ? 8.07-7.95 (m, 16H), 7.89-7.87 (d, J=7.5 Hz, 4H), 7.82-7.80 (d, J=8.2 Hz, 4H), 7.77-7.74 (m, 4H), 7.63-7.21 (m, 42H), 6.14 (t, J=9.9 Hz, 2H), 5.71-5.66 (m, 4H), 5.52 (d, J=9.5 Hz, 1H), 5.37-5.31 (m, 2H), 5.19-5.10 (m, 4H), 4.46-4.45 (m, 4H), 4.37-4.18 (m, 12H), 3.77 (d, J=6.9 Hz, 2H), 3.27-3.22 (m, 2H), 2.99 (t, J=9.2 Hz, 2H), 2.82 (t, J=8.6 Hz, 1H), 1.71-1.65 (m, 4H), 1.40-1.23 (m, 24H), 0.88 (t, J=7.2 Hz, 3H), 0.87 (t, J=7.2 Hz, 3H); .sup.13C NMR (100 MHz, CDCl.sub.3): ? 172.2, 171.9, 167.5, 166.2, 165.9, 165.8, 165.5, 165.1, 164.9, 134.0, 133.7, 133.5, 133.4, 133.2, 130.0, 129.9, 129.8, 129.7, 129.7, 129.5, 129.4, 129.3, 129.0, 128.9, 128.8, 128.7, 128.5, 128.4, 71.0, 128.3, 100.9, 95.6, 74.4, 73.2, 72.2, 71.1, 69.8, 69.1, 69.0, 67.9, 67.6, 63.2, 62.6, 48.2, 31.9, 29.6, 29.5, 29.3, 28.8, 28.7, 25.9, 22.7, 21.1, 14.2.

<3-4> Synthesis of TSM-E9

[0129] TSM-E9 was synthesized in 90% yield according to the general synthesis procedure for the deprotection reaction of Example 1-4. .sup.1H NMR (400 MHz, CD.sub.3OD): ? 5.17 (d, J=7.0 Hz, 2H), 4.37 (d, J=8.0 Hz, 2H), 4.32-4.21 (m, 4H), 4.00-3.96 (m, 1H), 3.89-3.88 (m, 2H), 3.83-3.79 (m, 2H), 3.75-3.70 (m, 4H), 3.61-3.42 (m, 10H), 3.37-3.30 (m, 4H), 3.20-3.16 (m, 4H), 1.68-1.62 (m, 4H), 1.36-1.22 (m, 20H), 0.88 (t, J=7.2 Hz, 3H), 0.87 (t, J=7.2 Hz, 3H); .sup.13C NMR (100 MHz, CD.sub.3OD): ? 173.2, 172.8, 169.0, 104.9, 104.7, 103.0, 81.3, 81.2, 77.7, 76.7, 75.1, 74.8, 74.7, 74.2, 71.5, 68.3, 68.9, 68.7, 62.8, 62.2, 32.0, 33.0, 33.1, 30.7, 30.5, 30.2, 30.0, 29.9, 27.1, 27.1,27.0, 23.9, 23.7, 14.6, 14.5; HRMS (FAB.sup.+): For C.sub.46H.sub.82N.sub.4O.sub.24 [M+Na].sup.+ 1097.5217, found 1097.5220.

Preparation Example 4

Synthesis of TSM-E10

<4-1> Synthesis of Compound 1d

[0130] Compound 1-d was synthesized in 50% yield according to Example 1-1. .sup.1H NMR (400 MHz, CDCl.sub.3): ? 4.41 (t, J=6.6 Hz, 4H), 1.81-1.77 (m, 4H), 1.34-1.27 (m, 28H), 0.88 (t, J=6.9 Hz, 6H); .sup.13C NMR (100 MHz, CDCl.sub.3): ? 172.7, 172.2, 69.6, 64.3, 32.0, 29.7, 29.6, 29.4, 28.6, 25.9, 22.8, 14.2.

<4-2> Synthesis of Compound 2d

[0131] Compound 2d was synthesized in 90% yield according to Example 1-2. .sup.1H NMR (400 MHz, CDCl.sub.3): ? 6.53 (d, J=8.0 Hz, 1H), 4.28-4.27 (m, 4H), 4.16-4.14 (m, 1H), 3.86-3.81 (m, 4H), 1.72-1.73 (m, 4H), 1.38-1.26 (m, 28H), 0.88 (t, J=7.0 Hz, 6H); .sup.13C NMR (100 MHz, CDCl.sub.3): ? 171.9, 171.4, 167.7, 67.9, 67.7, 62.1, 53.3, 31.9, 29.6, 29.4, 28.8, 25.9, 22.7, 14.1.

<4-3> Synthesis of TSM-E10a

[0132] TSM-E10a was synthesized in 82% yield according to the general glycosylation reaction procedure of Example 1-3. .sup.1H NMR (400 MHz, CDCl.sub.3): ? 8.06-7.93 (m, 16H), 7.88-7.86 (d, J=7.4 Hz, 4H), 7.81-7.79 (d, J=7.2 Hz, 4H), 7.76-7.72 (m, 4H), 7.63-7.22 (m, 42H), 6.12 (t, J=10.0 Hz, 2H), 5.71-5.60 (m, 4H), 5.49 (d, J=7.4 Hz, 2H), 5.35-5.29 (m, 2H), 5.16-5.08 (m, 4H), 4.61-4.50 (m, 4H), 4.39-4.17 (m, 12H), 3.76-3.74 (d, J=8.0 Hz, 2H), 3.25-3.19 (m, 2H), 2.97 (t, J=9.4 Hz, 2H), 2.80 (t, J=8.3 Hz, 1H), 1.70-1.66 (m, 4H), 1.26-1.23 (m, 28H), 0.88 (t, J=7.2 Hz, 3H), 0.87 (t, J=7.2 Hz, 3H); .sup.13C NMR (100 MHz, CDCl.sub.3): ? 172.3, 171.9, 166.2, 165.9, 165.6, 165.1, 164.9, 134.1, 133.7, 133.5, 133.4, 133.2, 130.0, 129.8, 129.7, 129.7, 129.5, 129.4, 129.3, 129.0, 128.9, 128.8, 128.7, 128.5, 128.4, 100.4, 95.7, 71.0, 71.4, 69.8, 69.1, 68.0, 67.7, 62.6, 32.0, 29.7, 29.4, 28.9, 28.8, 26.0, 22.8, 21.2, 14.3, 14.2.

<4-4> Synthesis of TSM-E10

[0133] TSM-E10 was synthesized in 91% yield according to the general synthesis procedure for the deprotection reaction of Example 1-4. .sup.1H NMR (400 MHz, CD.sub.3OD): ? 5.18 (d, J=7.0 Hz, 2H), 4.35 (d, J=8.0 Hz, 2H), 4.22-4.26 (m, 4H), 3.97-3.93 (m, 1H), 3.87-3.86 (m, 2H), 3.80-3.77 (m, 2H), 3.73-3.70 (m, 4H), 3.58-3.57 (m, 10H), 3.43-3.31 (m, 4H), 3.20-3.3.14 (m, 4H), 1.65-1.59 (m, 4H), 1.32-1.17 (m, 28H), 0.78 (t, J=7.0 Hz, 6H); .sup.13C NMR (100 MHz, CD.sub.3OD): ? 173.1, 169.0, 105.0, 104.8, 103.1, 81.4, 77.8, 76.7, 75.2, 74.7, 74.2, 71.6, 69.0, 68.8, 62.8, 62.2, 50.7, 33.2, 30.6, 30.5, 30.0, 27.1, 27.1, 23.9, 14.6; HRMS (FAB.sup.30): For C.sub.50H.sub.90N.sub.4O.sub.24 [M+Na].sup.+ 1153.5843, found 1153.5839.

Preparation Example 5

Synthesis of TSM-E11

<5-1> Synthesis of Compound 1e

[0134] Compound 1e was synthesized in 48% yield according to Example 1-1. .sup.1H NMR (400 MHz, CDCl.sub.3): ? 4.41 (t, J=6.6 Hz, 4H), 1.82-1.72 (m, 4H), 1.34-1.27 (m, 32H), 0.88 (t, J=6.9 Hz, 6H); .sup.13C NMR (100 MHz, CDCl.sub.3): ? 172.7, 172.2, 69.6, 32.0, 29.7, 29.6, 29.6, 29.3, 28.5, 25.8, 22.8, 14.2.

<5-2> Synthesis of Compound 2e

[0135] Compound 2e was synthesized in 88% yield according to Example 1-2. .sup.1H NMR (400 MHz, CDCl.sub.3): ? 6.19 (d, J=8.0 Hz, 1H), 4.32-4.26 (m, 4H), 4.16-4.14 (m, 1H), 3.93-3.85 (m, 4H), 1.74-1.73 (m, 4H), 1.36-1.26 (m, 32H), 0.88 (t, J=7.0 Hz, 6H); .sup.13C NMR (100 MHz, CDCl.sub.3): 172.1, 171.6, 168.0, 68.0, 67.9, 63.5, 53.2, 32.1, 29.8, 28.9, 26.0, 22.9, 14.3.

<5-3> Synthesis of TSM-E11a

[0136] TSM-E11a was synthesized in 84% yield according to the general glycosylation reaction procedure of Example 1-3. .sup.1H NMR (400 MHz, CDCl.sub.3): ? 8.05-7.93 (m, 16H), 7.88-7.86 (d, J=7.2 Hz, 4H), 7.81-7.79 (d, J=7.2 Hz, 4H), 7.75-7.72 (m, 4H), 7.63-7.22 (m, 42H), 6.11 (t, J=10.0 Hz, 2H), 5.68-5.61 (m, 4H), 5.48 (d, J=7.4 Hz, 2H), 5.35-5.28 (m, 2H), 5.17-5.14 (m, 4H), 4.66-4.52 (m, 4H), 4.33-4.10 (m, 12H), 3.74 (d, J=7.8 Hz, 2H), 3.25-3.17 (m, 2H), 2.93 (t, J=9.5 Hz, 2H), 2.79 (t, J=8.2 Hz, 1H), 1.72-1.62 (m, 4H), 1.44-1.29 (m, 32H), 0.89 (t, J=7.0 Hz, 6H); .sup.13C NMR (100 MHz, CDCl.sub.3): ? 172.3, 171.9, 166.2, 165.9, 165.6, 165.1, 164.9, 134.1, 133.7, 133.5, 133.4, 133.2, 130.0, 129.8, 129.7, 129.7, 129.5, 129.4, 129.3, 129.0, 128.9, 128.8, 128.7, 128.5, 128.4, 100.4, 95.7, 71.0, 71.4, 69.8, 69.1, 68.0, 67.7, 62.6, 32.0, 29.7, 29.4, 28.9, 28.8, 28.9, 26.0, 22.8, 21.2, 14.3, 14.2.

<5-4> Synthesis of TSM-E11

[0137] TSM-E11 was synthesized in 91% yield according to the general synthesis procedure for the deprotection reaction of Example 1-4. .sup.1H NMR (400 MHz, CD.sub.3OD): ? 5.16 (d, J=7.0 Hz, 2H), 4.39 (d, J=8.0 Hz, 2H), 4.29-4.16 (m, 4H), 3.96-3.92 (m, 1H), 3.85-3.84 (m, 2H), 3.80-3.76 (m, 2H), 3.71-3.69 (m, 4H), 3.56-3.39 (m, 10H), 3.34-3.26 (m, 4H), 3.18-3.3.12 (m, 4H), 1.64-1.58 (m, 4H), 1.32-1.17 (m, 32H), 0.77 (t, J=7.0 Hz, 6H); .sup.13C NMR (100 MHz, CD.sub.3OD): ? 173.1, 169.0, 105.0, 104.8, 103.1, 81.4, 77.8, 76.7, 75.2, 74.7, 74.2, 71.6, 69.0, 68.8, 62.8, 62.2, 50.7, 33.2, 30.6, 30.5, 30.0, 27.1, 27.1, 23.9, 14.6; HRMS (FAB.sup.+): For C.sub.52H.sub.94N.sub.4O.sub.24 [M+Na].sup.+ 1181.6159, found 1181.6162.

Preparation Example 6

Synthesis of TSM-T7

<6-1> Synthesis of Compound 3a

[0138] Compound 3a was synthesized in 85% yield according to Example 1-1. .sup.1H NMR (400 MHz, CDCl.sub.3): ? 4.42 (t, J=6.6 Hz, 4H), 1.81-1.77 (m, 4H), 1.44-1.30 (m, 16H), 0.89 (t, J=6.9 Hz, 6H); .sup.13C NMR (100 MHz, CDCl.sub.3): ? 172.7, 172.2, 69.6, 32.0, 29.6, 28.6, 25.9, 22.8, 14.2.

<6-2> Synthesis of Compound 4a

[0139] Compound 4a was synthesized in 90% yield according to Example 1-2. .sup.1H NMR (400 MHz, CDCl.sub.3): ? 6.27 (d, J=7.9 Hz, 1H), 4.14-4.09 (m, 4H), 3.90-3.81 (m, 4H), 3.04-3.01 (m, 4H), 1.68-1.65 (m, 4H), 1.40-1.26 (m, 16H), 0.88 (t, J=7.1 Hz, 6H); .sup.13C NMR (100 MHz, CDCl.sub.3): ? 180.5, 179.7, 162.5, 63.2, 32.0, 30.4, 30.2, 29.6, 29.5, 29.4, 29.1, 22.8, 14.2.

<6-3> Synthesis of TSM-T7a

[0140] TSM-T7a was synthesized in 85% yield according to the general glycosylation reaction procedure of Example 1-3. .sup.1H NMR (400 MHz, CDCl.sub.3): ? 8.27-7.91 (m, 12H), 7.90 (d, J=7.6 Hz, 4H), 7.83 (d, J=7.6 Hz, 4H), 7.78-7.75 (m, 4H), 7.69-7.19 (m, 42H), 6.17 (t, J=10.0 Hz, 2H), 5.74-5.66 (m, 4H), 5.48-5.34 (m, 4H), 5.16-5.13 (m, 4H), 4.72-4.61 (m, 4H), 4.37-4.23 (m, 10H), 3.79-3.76 (m 2H), 3.33-3.24 (m, 2H), 3.05-2.85 (m, 6H), 1.66-1.60 (m, 4H), 1.39-1.23 (m, 16H), 0.87 (t, J=6.9 Hz, 6H); .sup.13C NMR (100 MHz, CDCl.sub.3): ? 180.4, 179.7, 166.1, 165.8, 165.7, 165.4, 165.0, 164.8, 162.0, 133.4, 133.2, 129.9, 129.8, 129.7, 129.6, 129.4, 129.3, 129.2, 128.2, 128.9, 128.7, 128.6, 128.4, 128.3, 100.8, 95.6, 74.4, 71.9, 71.3, 69.7, 69.1, 68.8, 63.1, 62.5, 31.7, 30.1, 29.9, 29.3, 28.8, 28.7, 22.6, 14.1.

<6-4> Synthesis of TSM-T7

[0141] TSM-T7 was synthesized in 90% yield according to the general synthesis procedure for the deprotection reaction of Example 1-4. .sup.1H NMR (400 MHz, CD.sub.3OD): ? 5.18 (d, J=7.0 Hz, 2H), 4.38-4.36 (m, 3H), 3.93-3.90 (m, 1H), 3.84-3.76 (m, 4H), 3.72-3.66 (m, 4H), 3.55-3.39 (m, 10H), 3.33-3.26 (m, 4H), 3.17-3.12 (m, 6H), 2.94 (t, J=8.0 Hz, 4H), 1.57-1.53 (m, 4H), 1.31-1.18 (m, 16H), 0.77 (t, J=6.8 Hz, 6H); .sup.13C NMR (100 MHz, CD.sub.3OD): ? 181.7, 180.9, 164.0, 105.3, 105.1, 103.4, 81.8, 81.6, 78.1, 77.1, 75.5, 75.2, 75.1, 75.0, 74.6, 71.9, 69.8, 69.6, 63.2, 62.6, 62.4, 52.2, 33.5, 31.5, 31.3, 31.2, 30.9, 30.5, 30.4, 24.2, 15.0; HRMS (FAB.sup.+): For C.sub.44H.sub.78N.sub.4O.sub.22S.sub.2 [M+Na].sup.+ 1101.4447, found 1101.4453.

Preparation Example 7

Synthesis of TSM-T8

<7-1> Synthesis of Compound 3b

[0142] Compound 3b was synthesized in 84% yield according to Example 1-1. .sup.1H NMR (400 MHz, CDCl.sub.3): ? 4.42 (t, J=6.6 Hz, 4H), 1.81-1.77 (m, 4H), 1.44-1.30 (m, 20H), 0.89 (t, J=6.9 Hz, 6H); .sup.13C NMR (100 MHz, CDCl.sub.3): ? 172.7, 172.2, 69.6, 32.0, 29.7, 29.6, 28.6, 25.9, 22.8, 14.2.

<7-2> Synthesis of Compound 4b

[0143] Compound 4b was synthesized in 92% yield according to Example 1-2. .sup.1H NMR (400 MHz, CDCl.sub.3): ? 6.26 (d, J=7.8 Hz, 1H), 4.11-4.09 (m, 4H), 3.92-3.83 (m, 4H), 3.05-3.01 (m, 4H), 1.68-1.65 (m, 4H), 1.39-1.27 (m, 20H), 0.88 (t, J=7.0 Hz, 6H); .sup.13C NMR (100 MHz, CDCl.sub.3): ? 180.5, 179.7, 162.5, 63.2, 32.0, 30.4, 30.2, 29.6, 29.5, 29.4, 29.1, 22.8, 14.2.

<7-3> Synthesis of TSM-T8a

[0144] TSM-T8a was synthesized in 85% yield according to the general glycosylation reaction procedure of Example 1-3. .sup.1H NMR (400 MHz, CDCl.sub.3): ? 8.27-7.91 (m, 12H), 7.90 (d, J=7.6 Hz, 4H), 7.83 (d, J=7.6 Hz, 4H), 7.78-7.75 (m, 4H), 7.69-7.19 (m, 42H), 6.17 (t, J=10.0 Hz, 2H), 5.74-5.66 (m, 4H), 5.48-5.34 (m, 4H), 5.16-5.13 (m, 4H), 4.72-4.61 (m, 4H), 4.37-4.23 (m, 10H), 3.79-3.76 (m 2H), 3.33-3.24 (m, 2H), 3.05-2.85 (m, 6H), 1.66-1.60 (m, 4H), 1.39-1.23 (m, 20H), 0.87 (t, J=6.9 Hz, 6H); .sup.13C NMR (100 MHz, CDCl.sub.3): ? 180.4, 179.7, 166.1, 165.8, 165.7, 165.4, 165.0, 164.8, 162.0, 133.4, 133.2, 129.9, 129.8, 129.7, 129.6, 129.4, 129.3, 129.2, 128.2, 128.9, 128.7, 128.6, 128.4, 128.3, 100.8, 95.6, 74.4, 71.9, 71.3, 69.7, 69.1, 68.8, 63.1, 62.5, 31.7, 30.1, 29.9, 29.3, 28.8, 28.7, 22.6, 14.1.

<7-4> Synthesis of TSM-T8

[0145] TSM-T8 was synthesized in 90% yield according to the general synthesis procedure for the deprotection reaction of Example 1-4. .sup.1H NMR (400 MHz, CD.sub.3OD): ? 5.17 (d, J=7.0 Hz, 2H), 4.39-4.35 (m, 3H), 3.93-3.89 (m, 1H), 3.83-3.76 (m, 4H), 3.69-3.67 (m, 4H), 3.54-3.39 (m, 10H), 3.32-3.26 (m, 4H), 3.16-3.10 (m, 6H), 2.94 (t, J=8.0 Hz, 4H), 1.58-1.52 (m, 4H), 1.28-1.16 (m, 20H), 0.75 (t, J=7.2 Hz, 6H); .sup.13C NMR (100 MHz, CD.sub.3OD): ? 181.1, 180.8, 163.5, 105.3, 104.8, 104.6 102.9, 81.3, 81.2, 77.7, 76.6, 75.0, 74.8, 74.6, 74.1, 71.4, 69.3, 69.1, 62.7, 62.1, 52.2, 33.0, 31.0, 30.8, 30.6, 30.4, 30.3, 30.0, 29.9, 23.7, 14.5; HRMS (FAB.sup.+): For C.sub.46H.sub.82N.sub.4O.sub.22S.sub.2 [M+Na].sup.+ 1129.4760, found 1129.4757.

Preparation Example 8

Synthesis of TSM-T9

<8-1> Synthesis of Compound 3c

[0146] Compound 3c was synthesized in 86% yield according to Example 1-1. .sup.1H NMR (400 MHz, CDCl.sub.3): ? 4.42 (t, J=6.6 Hz, 4H), 1.81-1.78 (m, 4H), 1.43-1.30 (m, 24H), 0.89 (t, J=6.9 Hz, 6H); .sup.13C NMR (100 MHz, CDCl.sub.3): ? 172.7, 172.2, 69.6, 32.0, 29.6, 29.4, 28.6, 25.9, 22.8, 14.2.

<8-2> Synthesis of Compound 4c

[0147] Compound 4c was synthesized in 85% yield according to Example 1-2. .sup.1H NMR (400 MHz, CDCl.sub.3): ? 6.36 (d, J=8.0 Hz, 1H), 4.11-4.09 (m, 4H), 3.90-3.81 (m, 4H), 3.03-3.01 (m, 4H), 1.68-1.65 (m, 4H), 1.39-1.27 (m, 24H), 0.88 (t, J=7.0 Hz, 6H); .sup.13C NMR (100 MHz, CDCl.sub.3): ? 180.4, 179.6, 162.3, 62.6, 32.0, 30.4, 30.2, 29.7, 29.6, 29.5, 29.4, 29.1, 22.8, 14.2.

<8-3> Synthesis of TSM-T9a

[0148] TSM-T9a was synthesized in 86% yield according to the general synthesis procedure for the deprotection reaction of Example 1-3. .sup.1H NMR (400 MHz, CDCl.sub.3): ? 8.27-7.91 (m, 12H), 7.90 (d, J=7.6 Hz, 4H), 7.83 (d, J=7.6 Hz, 4H), 7.78-7.75 (m, 4H), 7.69-7.19 (m, 42H), 6.15 (t, J=10.0 Hz, 2H), 5.72-5.65 (m, 4H), 5.41-5.34 (m, 4H), 5.20-5.11 (m, 4H), 4.66-4.59 (m, 4H), 4.37-4.20 (m, 10H), 3.77-3.74 (m 2H), 3.28-3.23 (m, 2H), 3.05-2.95 (m, 6H), 1.65-1.59 (m, 4H), 1.30-1.21 (m, 24H), 0.87 (t, J=7.0 Hz, 6H); .sup.13C NMR (100 MHz, CDCl.sub.3): ? 180.4, 179.7, 166.1, 165.8, 165.7, 165.4, 165.0, 164.8, 162.0, 133.4, 133.2, 129.9, 129.8, 129.7, 129.6, 129.4, 129.3, 129.2, 128.2, 128.9, 128.7, 128.6, 128.4, 128.3, 100.8, 95.6, 74.4, 71.9, 71.3, 69.8, 69.1, 68.9, 63.2, 62.5, 32.0, 31.9, 30.2, 30.0, 29.6, 29.5, 29.3, 29.2, 28.9, 28.8, 28.7, 22.7, 14.2.

<8-4> Synthesis of TSM-T9

[0149] TSM-T9 was synthesized in 93% yield according to the general synthesis procedure for the deprotection reaction of Example 1-4. .sup.1H NMR (400 MHz, CD.sub.3OD): ? 5.16 (d, J=7.0 Hz, 2H), 4.34-4.09 (m, 3H), 3.94-3.90 (m, 1H), 3.83-3.76 (m, 4H), 3.72-3.69 (m, 4H), 3.57-3.39 (m, 10H), 3.27-3.26 (m, 4H), 3.17-3.12 (m, 6H), 2.93 (t, J=8.0 Hz, 4H), 1.60-1.50 (m, 4H), 1.30-1.16 (m, 24H), 0.76 (t, J=7.2 Hz, 6H); .sup.13C NMR (100 MHz, CD.sub.3OD): ? 181.7, 180.9, 164.0, 105.3, 105.1, 103.4, 81.8, 81.6, 78.1, 77.1, 75.5, 75.2, 75.0, 74.6, 71.9, 69.8, 69.6, 63.2, 62.6, 52.2, 33.5, 35.1, 31.3, 31.2. 30.9, 30.5, 30.1, 24.2, 14.9; HRMS (FAB.sup.+): For C.sub.48H.sub.86N.sub.4O.sub.22S.sub.2 [M+Na].sup.+ 1157.5073, found 1157.5068.

Preparation Example 9

Synthesis of TSM-T10

<9-1> Synthesis of Compound 3d

[0150] Compound 3d was synthesized in 83% yield according to Example 1-1. .sup.1H NMR (400 MHz, CDCl.sub.3): ? 4.42 (t, J=6.6 Hz, 4H), 1.81-1.78 (m, 4H), 1.43-1.30 (m, 28H), 0.89 (t, J=6.9 Hz, 6H); .sup.13C NMR (100 MHz, CDCl.sub.3): ? 172.7, 172.2, 69.6, 64.3, 32.0, 29.7, 29.6, 29.4, 28.6, 25.9, 22.8, 14.2.

<9-2> Synthesis of Compound 4d

[0151] Compound 4d was synthesized in 85% yield according to Example 1-2. .sup.1H NMR (400 MHz, CDCl.sub.3): ? 6.0 (d, J=8.0 Hz, 1H), 4.10-4.08 (m, 4H), 3.91-3.89 (m, 4H), 3.05-3.01 (m, 4H), 1.68-1.65 (m, 4H), 1.39-1.27 (m, 28H), 0.88 (t, J=7.0 Hz, 6H); .sup.13C NMR (100 MHz, CDCl.sub.3): ? 180.5, 179.7, 162.6, 63.2, 34.2, 32.1, 32.0, 30.4, 30.2, 29.7, 29.6, 29.5, 29.4, 29.2, 29.1, 28.5, 24.8, 22.8, 14.2.

<9-3> Synthesis of TSM-T10a

[0152] TSM-T10a was synthesized in 85% yield according to the general glycosylation reaction procedure of Example 1-3. .sup.1H NMR (400 MHz, CDCl.sub.3): ? 8.27-7.91 (m, 12H), 7.90 (d, J=7.6 Hz, 4H), 7.83 (d, J=7.6 Hz, 4H), 7.78-7.75 (m, 4H), 7.69-7.19 (m, 42H), 6.15 (t, J=10.0 Hz, 2H), 5.72-5.65 (m, 4H), 5.41-5.34 (m, 4H), 5.20-5.11 (m, 4H), 4.66-4.59 (m, 4H), 4.37-4.20 (m, 10H), 3.77-3.74 (m 2H), 3.28-3.23 (m, 2H), 3.05-2.95 (m, 6H), 1.65-1.59 (m, 4H), 1.30-1.21 (m, 28H), 0.87 (t, J=7.0 Hz, 6H); .sup.13C NMR (100 MHz, CDCl.sub.3): ? 180.5, 179.8, 166.1, 165.8, 165.7, 165.4, 165.0, 164.8, 162.0, 133.4, 133.2, 129.9, 129.8, 129.7, 129.6, 129.4, 129.3, 129.2, 128.2, 128.9, 128.7, 128.6, 128.4, 128.3, 100.8, 95.6, 74.5, 72.5, 71.9, 71.3, 69.8, 69.1, 69.0, 63.2, 62.6, 32.0, 31.9, 30.3, 30.1, 30.0, 29.7, 29.4, 29.5, 29.3, 29.2, 28.9, 28.8, 28.7, 22.8, 14.2.

<9-4> Synthesis of TSM-T10

[0153] TSM-T10 was synthesized in 92% yield according to the general synthesis procedure for the deprotection reaction of Example 1-4. .sup.1H NMR (400 MHz, CD.sub.3OD): ? 5.17 (d, J=7.0 Hz, 2H), 4.34-4.30 (m, 3H), 3.94-3.90 (m, 1H), 3.83-3.76 (m, 4H), 3.72-3.69 (m, 4H), 3.57-3.39 (m, 10H), 3.27-3.26 (m, 4H), 3.17-3.12 (m, 6H), 2.93 (t, J=8.0 Hz, 4H), 1.60-1.50 (m, 4H), 1.30-1.16 (m, 28H), 0.76 (t, J=7.2 Hz, 6H); .sup.13C NMR (100 MHz, CD.sub.3OD): ? 181.6, 180.8, 163.7, 104.9, 104.7, 103.0, 81.4, 77.7, 76.7, 75.1, 74.9, 74.7, 74.2, 71.5, 62.8, 62.2, 33.2, 30.8, 30.6, 30.5, 30.1, 23.9, 14.6; HRMS (FAB.sup.+): For C.sub.50H.sub.90N.sub.4O.sub.22S.sub.2 [M+Na].sup.+ 1185.5386, found 1185.5392.

Preparation Example 10

Synthesis of TSM-T11

<10-1> Synthesis of Compound 3e

[0154] Compound 3e was synthesized in 82% yield according to Example 1-1. .sup.1H NMR (400 MHz, CDCl.sub.3): ? 4.42 (t, J=6.8 Hz, 4H), 1.81-1.78 (m, 4H), 1.43-1.30 (m, 32H), 0.89 (t, J=6.9 Hz, 6H); .sup.13C NMR (100 MHz, CDCl.sub.3): ? 172.7, 172.2, 69.6, 32.0, 29.7, 29.6, 29.6, 29.3, 28.5, 25.8, 22.8, 14.2.

<10-2> Synthesis of Compound 4e

[0155] Compound 4e was synthesized in 84% yield according to Example 1-2. .sup.1H NMR (400 MHz, CDCl.sub.3): ? 6.38 (d, J=8.1 Hz, 1H), 4.10-4.08 (m, 4H), 3.91-3.89 (m, 4H), 3.05-3.01 (m, 4H), 1.68-1.65 (m, 4H), 1.39-1.27 (m, 32H), 0.88 (t, J=7.0 Hz, 6H); .sup.13C NMR (100 MHz, CDCl.sub.3): ? 180.5, 179.7, 162.6, 63.2, 34.2, 32.1, 32.0, 30.4, 30.2, 29.7, 29.6, 29.5, 29.4, 29.2, 29.1, 28.5, 24.8, 22.8, 14.2.

<10-3> Synthesis of TSM-T11a

[0156] TSM-T11a was synthesized in 85% yield according to the general glycosylation reaction procedure of Example 1-3. .sup.1H NMR (400 MHz, CDCl.sub.3): ? 8.27-7.91 (m, 12H), 7.90 (d, J=7.6 Hz, 4H), 7.83 (d, J=7.6 Hz, 4H), 7.78-7.75 (m, 4H), 7.69-7.19 (m, 42H), 6.15 (t, J=10.0 Hz, 2H), 5.72-5.65 (m, 4H), 5.41-5.34 (m, 4H), 5.20-5.11 (m, 4H), 4.66-4.59 (m, 4H), 4.37-4.20 (m, 10H), 3.77-3.74 (m 2H), 3.28-3.23 (m, 2H), 3.05-2.95 (m, 6H), 1.65-1.59 (m, 4H), 1.30-1.21 (m, 32H), 0.87 (t, J=7.0 Hz, 6H); .sup.13C NMR (100 MHz, CDCl.sub.3): ? 180.5, 179.8, 166.1, 165.8, 165.7, 165.4, 165.0, 164.8, 162.0, 133.4, 133.2, 129.9, 129.8, 129.7, 129.6, 129.4, 129.3, 129.2, 128.2, 128.9, 128.7, 128.6, 128.4, 128.3, 100.8, 95.6, 74.5, 72.3, 71.9, 71.3, 69.8, 69.1, 69.0, 68.9, 63.2, 62.6, 32.0, 31.9, 30.3, 30.1, 30.0, 29.7, 29.4, 29.5, 29.3, 29.2, 28.9, 28.8, 28.7, 22.8, 14.2.

<10-4>Synthesis of TSM-T11

[0157] TSM-T11 was synthesized in 90% yield according to the general synthesis procedure for the deprotection reaction of Example 1-4. .sup.1H NMR (400 MHz, CD.sub.3OD): ? 5.16 (d, J=7.0 Hz, 2H), 4.39-4.34 (m, 3H), 3.95-3.91 (m, 1H), 3.82-3.77 (m, 4H), 3.72-3.69 (m, 4H), 3.57-3.40 (m, 10H), 3.35-3.29 (m, 4H), 3.18-3.14 (m, 6H), 2.95 (t, J=8.0 Hz, 4H), 1.62-1.52 (m, 4H), 1.30-1.17 (m, 32H), 0.78 (t, J=7.2 Hz, 6H); .sup.13C NMR (100 MHz, CD.sub.3OD): ? 180.5, 162.9, 104.6, 103.0, 81.3, 77.9, 76.7, 75.1, 74.8, 74.7, 74.2, 71.5, 69.1, 62.8, 62.2, 31.2, 31.1, 31.0, 30.9, 30.8, 30.6, 30.5, 30.1, 23.9, 14.6; HRMS (FAB.sup.+): For C.sub.52H.sub.94N.sub.4O.sub.22S.sub.2 [M+Na].sup.+ 1213.5699, found 1213.5702.

EXAMPLE 2

Synthesis Method of TTGs

[0158] FIG. 2 illustrates the synthesis scheme of TTGs. 20 types of compounds of TTGs were synthesized by the following synthesis methods of <2-1> to <2-4>.

<2-1>General Procedure for the Synthesis of 2-chloro-4,6-dialkylated-1,3,5-triazine (Step i of FIG. 2)

[0159] A mixture of 2,4,6-trichloro-1,3,5-triazine (3.01 mmol) and NaHCO.sub.3 (7.26 mmol) was stirred in acetone (10 mL) for 10 minutes. Each thiol (RSH) (6.0 mmol) or alkyl amine dissolved in acetone was added dropwise for 30 minutes. The resulting reaction mixture was kept at room temperature for 1 hr. The reaction mixture was extracted with CHCl.sub.3 and water, and the organic layer was dried over anhydrous Na.sub.2SO.sub.4. The oily residue obtained after removal of solvent was subjected to column chromatography purification to obtain target Compound 1 or 3.

<2-2> General Synthesis Procedure for the Coupling Reactions of the Resulting Dialkylated Triazine Derivatives With 2-amino-1,3-propanediol (Step ii of FIG. 2)

[0160] To a dry flask solution of 2-chloro-4,6-dialkylated-1,3,5-triazine (1.00 g, 5.42 mmol) dissolved in THF (50 mL), tris(hydroxymethyl)aminomethane (1.5 equiv.) and diisopropylethylamine (DIPEA) were added under nitrogen. After the addition, the temperature was gradually increased to 100? C., and the mixture was further stirred for 34 hours. The reaction mixture was diluted with water and then extracted with ethyl acetate. The organic layer was washed 1.0 M HCl and brine and dried over anhydrous Na.sub.2SO.sub.4. After concentration of the ethyl acetate solution, the residue was purified by flash column chromatography (EtOAc/hexane) to obtain desired target Compound 2 or 4.

<2-3> General Synthesis Procedure for Glycosylation Reaction (Step iii of FIG. 2)

[0161] This reaction was performed according to the synthesis method (Nat. Methods 2010, 7, 1003.) of Chae, P. S. et al. with some modifications. Briefly, a mixture of a dialkylated tri-ol derivative (1 equiv., 250 mg), AgOTf (2.4 equiv.), 2,4,6-collidine (1 equiv.) in anhydrous CH.sub.2Cl.sub.2 (40 mL) was stirred at ?45? C. Then, perbenzoylated maltosylbromide (2.4 equiv.) dissolved in CH.sub.2Cl.sub.2 (10 mL) was added dropwise over 0.5 hr into this suspension. The reaction proceeded with continuous stirring at ?45? C. for 30 minutes. Then, the reaction mixture was warmed to 0? C. and stirring was continued for 1 hour. After completion of the reaction (as detected by TLC), pyridine (1.0 mL) was added to the reaction mixture. The reaction mixture was diluted with CH.sub.2Cl.sub.2 (40 mL) before being filtered over Celite. The filtrate was washed successively with a 1.0 M aqueous Na.sub.2S.sub.2O.sub.3 solution (40 mL), a 0.1 M aqueous HCl solution (40 mL), and brine (2?40 mL). Then, the organic layer was dried with anhydrous Na.sub.2SO.sub.4 and the solvent was removed by rotary evaporation. The resulting residue was purified by silica gel column chromatography (EtOAc/hexane) to obtain the glycosylated target compound.

<2-4> General Synthesis Procedure for Deprotection Reaction (Step iv of FIG. 2)

[0162] This reaction followed the synthesis method (Nat. Methods 2010, 7, 1003.) of Chae, P. S. aureus et al. The de-O-benzoylation was performed under Zemplen's condition. An O-protected compound was dissolved in anhydrous CH.sub.2Cl.sub.2 and then MeOH was added slowly thereto until persistent precipitation appeared. A methanolic solution of 0.5 M NaOMe was added to the reaction mixture such that the final concentration of NaOMe was 0.05 M. The reaction mixture was stirred for 14 hr at room temperature. After completion of the reaction, the reaction mixture was neutralized using Amberlite IR-120 (H.sup.+ form) resin. The resin was removed by filtration and washed with MeOH and the solvent was removed from the filtrate in vacuo. The residue was purified by silica gel column chromatography (CH.sub.2Cl.sub.2/MeOH) to obtain the target compound.

Preparation Example 11

Synthesis of TTG-T7

<11-1> Synthesis of Compound 1a

[0163] Compound 1a was synthesized in 82% yield according to Example 2-1. .sup.1H NMR (400 MHz, CDCl.sub.3): ? 4.42 (t, J=6.6 Hz, 4H), 1.81-1.77 (m, 4H), 1.44-1.30 (m, 16H), 0.89 (t, J=6.9 Hz, 6H); .sup.13C NMR (100 MHz, CDCl.sub.3): ? 172.7, 172.2, 69.6, 32.0, 29.7, 29.6, 28.6, 25.9, 22.8, 14.2.

<11-2> Synthesis of Compound 2a

[0164] Compound 2a was synthesized in 80% yield according to Example 2-2. .sup.1H NMR (400 MHz, CDCl.sub.3): ? 5.07 (b, 3H), 3.69 (s, 6H), 3.04-3.01(m, 4H), 1.67-1.66 (m, 4H), 1.39-1.28 (m, 16H), 0.88 (t, J=7.04 Hz, 6H) .sup.13C NMR (100 MHz, CDCl.sub.3): ? 180.5, 179.7, 162.5, 63.2, 32.0, 30.4, 30.2, 29.6, 29.5, 29.4, 29.1, 22.8, 14.2.

<11-3> Synthesis of TTG-T7a

[0165] TTG-T7a was synthesized in 85% yield according to the general glycosylation reaction procedure of Example 2-3. .sup.1H NMR (400 MHz, CDCl.sub.3): ? 8.08-8.01 (m, 12H), 7.91 (d, J=7.2 Hz, 6H), 7.83 (d, J=8 Hz 6H), 7.71-7.26 (m, 46H), 5.97 (s, 1H), 5.60 (t, J=8 Hz, 3H), 5.44 (t, J=12 Hz, 3H), 5.29-5.2 (m, 4H), 4.50-4.37 (m, 8H), 4.10-3.90 (m, 4H), 3.57-3.50 (m, 5H), 2.94-2.90 (m 4H), 1.59-1.55 (m, 4H), 1.26-1.21 (m, 16H), 0.88 (t, J=7.4 Hz 6H), .sup.13C NMR (100 MHz, CDCl.sub.3): ? 166.2, 165.3, 165.1, 164.7, 162.6, 133.8, 133.6, 133.4, 133.3, 130.1, 129.9, 129.8, 129.6, 129.5, 129.2, 128.9, 128.8, 128.6, 128.4, 101.5, 72.6, 72.0, 71.8, 69.6, 68.3, 63.2, 59.8, 31.8, 29.2, 28.9, 22.7, 14.2.

<11-4> Synthesis of TTG-T7

[0166] TTG-T7 was synthesized in 84% yield according to the general synthesis procedure for the deprotection reaction of Example 2-4. .sup.1H NMR (400 MHz, CD.sub.3OD): ? 4.35 (d, J=8 Hz, 3H), 4.19 (d, J=8 Hz, 3H), 3.87 (d, J=12 Hz, 3H), 3.75-3.72 (m, 3H), 3.59-3.55 (m, 3H), 3.25-3.16 (m, 10H), 3.08 (t, J=8 Hz, 3H), 2.97-2.94 (m, 4H), 1.60-1.53 (m, 4H), 1.31-1.19 (m, 16H), 0.78 (t, J=8.0 Hz 6H) .sup.13C NMR (100 MHz, CD.sub.3OD): ? 163.9, 105.6, 78.2, 78.21, 75.3, 71.7, 70.3, 62.8, 61.5, 33.3, 31.2, 31.0, 30.7, 30.5, 30.3, 24.0, 14.8.

Preparation Example 12

Synthesis of TTG-T8

<12-1> Synthesis of Compound 1b

[0167] Compound 1b was synthesized in 84% yield according to Example 2-1. .sup.1H NMR (400 MHz, CDCl.sub.3): ? 4.42 (t, J=6.6 Hz, 4H), 1.81-1.78 (m, 4H), 1.43-1.30 (m, 20H), 0.89 (t, J=6.9 Hz, 6H); .sup.13C NMR (100 MHz, CDCl.sub.3): ? 172.7, 172.2, 69.6, 32.0, 29.6, 29.4, 28.6, 25.9, 22.8, 14.2.

<12-2> Synthesis of Compound 2b

[0168] Compound 2b was synthesized in 82% yield according to Example 2-2. .sup.1H NMR (400 MHz, CDCl.sub.3): ? 4.92 (b, 3H), 3.69 (s, 6H), 3.04-3.01(m, 4H), 1.70-1.66 (m, 4H), 1.28-1.26 (m, 20H), 0.88 (t, J=7.4 Hz, 6H) .sup.13C NMR (100 MHz, CDCl.sub.3): ? 180.5, 179.7, 162.5, 63.2, 32.0, 30.4, 30.2, 29.6, 29.5, 29.4, 29.1, 22.8, 14.2.

<12-3> Synthesis of TTG-T8a

[0169] TTG-T8a was synthesized in 86% yield according to the general glycosylation reaction procedure of Example 2-3. .sup.1H NMR (400 MHz, CDCl.sub.3): ? 8.08-8.01 (m, 12H), 7.91 (d, J=7.2 Hz, 6H), 7.83 (d, J=8 Hz, 6H), 7.71-7.26 (m, 46H), 5.97 (s, 1H), 5.60 (t, J=8 Hz, 3H), 5.44 (t, J=12 Hz, 3H), 5.29-5.2 (m, 4H), 4.50-4.37 (m, 8H), 4.10-3.90 (m, 4H), 3.57-3.50 (m, 5H), 2.94-2.90 (m 4H), 1.59-1.55 (m, 4H), 1.26-1.21 (m, 20H), 0.88 (t, J=7.4 Hz, 6H), .sup.13C NMR (100 MHz, CDCl.sub.3): ? 166.1, 165.1, 164.6, 162.5, 133.8, 133.5, 133.3, 133.2, 130.0, 129.8, 129.7, 129.6, 129.1, 128.8, 128.5, 128.4, 101.5, 101.4, 72.5, 71.9, 71.8, 69.5, 68.2, 63.2, 59.8, 31.9, 29.6, 29.4, 29.2, 29.0, 22.7, 22.1, 14.2.

<12-4> Synthesis of TTG-T8

[0170] TTG-T8 was synthesized in 85% yield according to the general synthesis procedure for the deprotection reaction of Example 2-4. .sup.1H NMR (400 MHz, CD.sub.3OD): ? 4.35 (d, J=8 Hz, 3H), 4.19 (d, J=8 Hz, 3H), 3.87 (d, J=12 Hz, 3H), 3.75-3.72 (m, 3H), 3.59-3.55 (m, 3H), 3.25-3.16 (m, 10H), 3.08 (t, J=8 Hz, 3H), 2.97-2.94 (m, 4H), 1.60-1.53 (m, 4H), 1.31-1.19 (m, 20H), 0.78 (t, J=8.0 Hz, 6H) .sup.13C NMR (100 MHz, CD.sub.3OD): ? 163.9, 105.6, 78.2, 78.21, 75.3, 71.7, 70.3, 62.8, 61.5, 33.3, 31.2, 31.0, 30.7, 30.5, 30.3, 24.0, 14.8.

Preparation Example 13

Synthesis of TTG-T9

<13-1> Synthesis of Compound 1c

[0171] Compound 1c was synthesized in 85% yield according to Example 2-1. .sup.1H NMR (400 MHz, CDCl.sub.3): ? 4.42 (t, J=6.6 Hz, 4H), 1.81-1.78 (m, 4H), 1.43-1.30 (m, 24H), 0.89 (t, J=6.9 Hz, 6H); .sup.13C NMR (100 MHz, CDCl.sub.3): ? 172.7, 172.2, 69.6, 64.3, 32.0, 29.7, 29.5, 29.4, 28.5, 25.9, 22.7, 14.2.

<13-2> Synthesis of Compound 2c

[0172] Compound 2c was synthesized in 83% yield according to Example 2-2. .sup.1H NMR (400 MHz, CDCl.sub.3): ? 4.92 (b, 3H), 3.69 (s, 6H), 3.04-3.01(m, 4H), 1.70-1.66 (m, 4H), 1.28-1.26 (m, 24H), 0.88 (t, J=7.04 Hz, 6H) .sup.13C NMR (100 MHz, CDCl.sub.3): ? 180.5, 179.7, 162.5, 63.2, 32.0, 30.4, 30.2, 29.6, 29.5, 29.4, 29.1, 22.8, 14.2.

<13-3> Synthesis of TTG-T9a

[0173] TTG-T9a was synthesized in 85% yield according to the general glycosylation reaction procedure of Example 2-3. .sup.1H NMR (400 MHz, CDCl.sub.3): ? 8.08-8.01 (m, 12H), 7.91 (d, J=7.2 Hz, 6H), 7.83 (d, J=8 Hz 6H), 7.71-7.26 (m, 46H), 5.97 (s, 1H), 5.60 (t, J=8 Hz, 3H), 5.44 (t, J=12 Hz, 3H), 5.29-5.2 (m, 4H), 4.50-4.37 (m, 8H), 4.10-3.90 (m, 4H), 3.57-3.50 (m, 5H), 2.94-2.90 (m 4H), 1.59-1.55 (m, 4H), 1.26-1.21 (m, 24H), 0.88 (t, J=7.4 Hz 6H), .sup.13C NMR (100 MHz, CDCl.sub.3): ? 166.2, 165.1, 164.5, 162.4, 133.8, 133.5, 133.3, 133.1, 130.0, 129.8, 129.7, 129.6, 129.1, 128.8, 128.5, 128.4, 101.5, 101.4, 72.4, 71.9, 71.8, 69.5, 68.5, 63.2, 59.6, 31.8, 29.6, 29.4, 29.2, 29.0, 22.7, 22.1, 14.2.

<13-4> Synthesis of TTG-T9

[0174] TTG-T9 was synthesized in 84% yield according to the general synthesis procedure for the deprotection reaction of Example 2-4. .sup.1H NMR (400 MHz, CD.sub.3OD): ? 4.35 (d, J=8 Hz, 3H), 4.19 (d, J=8 Hz, 3H), 3.87 (d, J=12 Hz, 3H), 3.75-3.72 (m, 3H), 3.59-3.55 (m, 3H), 3.25-3.16 (m, 10H), 3.08 (t, J=8 Hz, 3H), 2.97-2.94 (m, 4H), 1.60-1.53 (m, 4H), 1.31-1.19 (m, 24H), 0.78 (t, J=8.0 Hz, 6H) .sup.13C NMR (100 MHz, CD.sub.3OD): ? 163.9, 105.6, 78.2, 78.21, 75.3, 71.7, 70.3, 62.8, 61.5, 33.3, 31.2, 31.0, 30.7, 30.5, 30.3, 24.0, 14.8.

Preparation Example 14

Synthesis of TTG-T10

<14-1> Synthesis of Compound 1d

[0175] Compound 1d was synthesized in 85% yield according to Example 2-1. .sup.1H NMR (400 MHz, CDCl.sub.3): ? 4.42 (t, J=6.6 Hz, 4H), 1.81-1.78 (m, 4H), 1.43-1.30 (m, 32H), 0.89 (t, J=6.9 Hz, 6H); .sup.13C NMR (100 MHz, CDCl.sub.3): ? 172.7, 172.2, 69.6, 32.0, 29.8, 29.7, 29.6, 29.3, 28.5, 25.8, 22.8, 14.2.

<14-2> Synthesis of Compound 2d

[0176] Compound 2d was synthesized in 85% yield according to Example 2-2. .sup.1H NMR (400 MHz, CDCl.sub.3): ? 4.92 (b, 3H), 3.69 (s, 6H), 3.04-3.01(m, 4H), 1.70-1.66 (m, 4H), 1.28-1.26 (m, 28H), 0.88 (t, J=7.04 Hz, 6H) .sup.13C NMR (100 MHz, CDCl.sub.3): ? 180.5, 179.6, 162.5, 63.1, 32.0, 30.4, 30.2, 29.6, 29.5, 29.4, 29.1, 22.7, 14.2.

<14-3> Synthesis of TTG-T10a

[0177] TTG-T10a was synthesized in 84% yield according to the general glycosylation reaction procedure of Example 2-3. .sup.1H NMR (400 MHz, CDCl.sub.3): ? 8.08-8.01 (m, 12H), 7.91 (d, J=7.2 Hz, 6H), 7.83 (d, J=8 Hz, 6H), 7.71-7.26 (m, 46H), 5.97 (s, 1H), 5.60 (t, J=8 Hz, 3H), 5.44 (t, J=12 Hz, 3H), 5.29-5.2 (m, 4H), 4.50-4.37 (m, 8H), 4.10-3.90 (m, 4H), 3.57-3.50 (m, 5H), 2.94-2.90 (m 4H), 1.59-1.55 (m, 4H), 1.26-1.21 (m, 28H), 0.88 (t, J=7.4 Hz, 6H), .sup.13C NMR (100 MHz, CDCl.sub.3): ? 166.1, 165.1, 164.6, 162.5, 133.8, 133.5, 133.3, 133.2, 130.0, 129.8, 129.7, 129.6, 129.1, 128.8, 128.5, 128.4, 101.5, 101.4, 72.5, 71.9, 71.8, 69.5, 68.2, 63.2, 60.4, 59.8, 53.5, 31.9, 29.6, 29.4, 29.2, 29.0, 22.7, 22.1, 14.2.

<14-4> Synthesis of TTG-T10

[0178] TTG-T10 was synthesized in 83% yield according to the general synthesis procedure for the deprotection reaction of Example 2-4. .sup.1H NMR (400 MHz, CD.sub.3OD): ? 4.33 (d, J=12 Hz, 3H), 4.18 (d, J=8 Hz, 3H), 3.87 (d, J=8 Hz, 3H), 3.77-3.71 (m, 3H), 3.58-3.54 (m, 3H), 3.24-3.15 (m, 10H), 3.07 (t, J=8 Hz, 3H), 2.95-2.93 (m, 4H), 1.59-1.53 (m, 4H), 1.30-1.17 (m, 28H), 0.77 (t, J=8.0 Hz, 6H) .sup.13C NMR (100 MHz, CD.sub.3OD): ? 163.5, 105.3, 78.0, 78.01, 75.2, 71.5, 70.1, 62.6, 61.3, 33.2, 30.8, 30.7, 30.6, 23.8, 14.6.

Preparation Example 15

Synthesis of TTG-T11

<15-1> Synthesis of Compound 1e

[0179] Compound 1e was synthesized in 86% yield according to Example 2-1. .sup.1H NMR (400 MHz, CDCl.sub.3): ? 4.42 (t, J=6.6 Hz, 4H), 1.81-1.78 (m, 4H), 1.43-1.30 (m, 32H), 0.89 (t, J=6.9 Hz, 6H); .sup.13C NMR (100 MHz, CDCl.sub.3): ? 172.7, 172.2, 69.6, 32.0, 29.8, 29.7, 29.6, 29.3, 28.5, 25.8, 22.8, 14.2.

<15-2> Synthesis of Compound 2e

[0180] Compound 2e was synthesized in 85% yield according to Example 2-2. .sup.1H NMR (400 MHz, CDCl.sub.3): ? 5.19 (b, 3H), 3.69 (s, 6H), 3.04-3.01 (m, 4H), 1.67-1.66 (m, 4H), 1.28-1.26 (m, 32H), 0.88 (t, J=7.04 Hz, 6H) .sup.13C NMR (100 MHz, CDCl.sub.3): ? 180.4, 179.5, 162.3, 62.5, 53.0, 30.0, 30.4, 30.2, 29.8, 29.7, 29.5, 29.4, 29.1, 22.8, 14.2.

<15-3> Synthesis of TTG-T11a

[0181] TTG-T11a was synthesized in 86% yield according to the general glycosylation reaction procedure of Example 2-3. .sup.1H NMR (400 MHz, CDCl.sub.3): ? 8.08-8.01 (m, 12H), 7.91 (d, J=7.2 Hz, 6H), 7.83 (d, J=8.0 Hz, 6H), 7.71-7.26 (m, 46H), 5.97 (s, 1H), 5.61 (t, J=8 Hz, 3H), 5.45 (t, J=12.0 Hz, 3H), 5.29-5.2 (m, 4H), 4.51-4.37 (m, 8H), 4.10-3.90 (m, 4H), 3.57-3.50 (m, 5H), 2.94-2.90 (m 4H), 1.58-1.55 (m, 4H), 1.26-1.21 (m, 32H), 0.88 (t, J=7.4 Hz, 6H), .sup.13C NMR (100 MHz, CDCl.sub.3): ? 166.3, 165.2, 164.5, 162.4, 133.8, 133.5, 133.3, 133.2, 130.0, 129.9, 129.7, 129.6, 129.1, 128.8, 128.5, 128.4, 101.5, 101.4, 92.7, 73.1, 72.8, 72.1, 71.9, 71.9, 69.4, 68.2, 63.2, 60.4, 59.8, 53.5, 48.4, 31.9, 30.2, 29.7, 29.6, 29.5, 29.2, 29.0, 22.7, 22.1, 14.2.

<15-4> Synthesis of TTG-T11

[0182] TTG-T11 was synthesized in 85% yield according to the general synthesis procedure for the deprotection reaction of Example 2-4. .sup.1H NMR (400 MHz, CD.sub.3OD): ? 4.34 (d, J=8.0 Hz, 3H), 4.19 (d, J=8.0 Hz 3H), 3.88 (d, J=12 Hz, 3H), 3.76-3.73 (m, 3H), 3.6-3.55 (m, 3H), 3.25-3.16 (m, 10H), 3.08 (t, J=8.0 Hz, 3H), 2.97-2.94 (m, 4H), 1.60-1.54 (m, 4H), 1.32-1.17 (m, 32H), 0.78 (t, J=8.0 Hz, 6H) .sup.13C NMR (100 MHz, CD.sub.3OD): ? 163.9, 105.6, 78.2, 78.21, 75.3, 71.7, 70.3, 62.8, 61.5, 33.3, 31.2, 31.0, 30.7, 30.5, 30.3, 24.0, 14.8.

Preparation Example 16

Synthesis of TTG-T12

<16-1> Synthesis of Compound 1f

[0183] Compound if was synthesized in 84% yield according to Example 2-1. .sup.1H NMR (400 MHz, CDCl.sub.3): ? 4.43 (t, J=6.6 Hz, 4H), 1.81-1.78 (m, 4H), 1.43-1.30 (m, 36H), 0.89 (t, J=6.9 Hz, 6H); .sup.13C NMR (100 MHz, CDCl.sub.3): ? 172.6, 172.2, 69.7, 32.0, 29.7, 29.8, 29.6, 29.3, 28.5, 25.7, 22.8, 14.2.

<16-2> Synthesis of Compound 2f

[0184] Compound 2f was synthesized in 84% yield according to Example 2-2. .sup.1H NMR (400 MHz, CDCl.sub.3): ? 4.92 (b, 3H), 3.69 (s, 6H), 3.04-3.01(m, 4H), 1.70-1.66 (m, 4H), 1.28-1.26 (m, 36H), 0.88 (t, J=7.4 Hz, 6H) .sup.13C NMR (100 MHz, CDCl.sub.3): ? 180.5, 179.7, 162.5, 63.2, 32.0, 30.4, 30.2, 29.6, 29.5, 29.4, 29.1, 22.8, 14.2.

<16-3> Synthesis of TTG-T12a

[0185] TTG-T12a was synthesized in 85% yield according to the general glycosylation reaction procedure of Example 2-3. .sup.1H NMR (400 MHz, CDCl.sub.3): ? 8.08-8.01 (m, 12H), 7.91 (d, J=7.2 Hz, 6H), 7.83 (d, J=8.0 Hz, 6H), 7.71-7.26 (m, 46H), 5.95 (s, 1H), 5.61 (t, J=8.0 Hz 3H), 5.46 (t, J=12.0 Hz, 3H), 5.28-5.24 (m, 4H), 4.49-4.39 (m, 8H), 4.10-3.90 (m, 4H), 3.51-3.49 (m, 5H), 2.95-2.90 (m 4H), 1.59-1.57 (m, 4H), 1.29-1.21 (m, 36H), 0.87 (t, J=7.4 Hz, 6H), .sup.13C NMR (100 MHz, CDCl.sub.3): ? 166.1, 165.2, 164.6, 162.4, 133.8, 133.5, 133.3, 133.2, 130.0, 129.8, 129.7, 129.6, 129.1, 128.8, 128.5, 128.4, 101.5, 101.4, 92.7, 72.6, 72.0, 71.4, 69.6, 68.2, 63.2, 59.9, 48.4, 31.9, 32.0, 29.6, 29.3, 29.1, 22.8, 14.2.

<16-4> Synthesis of TTG-T12

[0186] TTG-T12 was synthesized in 88% yield according to the general synthesis procedure for the deprotection reaction of Example 2-4. .sup.1H NMR (400 MHz, CD.sub.3OD): ? 4.35 (d, J=8.0 Hz, 3H), 4.19 (d, J=8.0 Hz, 3H), 3.88 (d, J=12.0 Hz, 3H), 3.76-3.73 (m, 3H), 3.6-3.55 (m, 3H), 3.25-3.16 (m, 10H), 3.08 (t, J=8.0 Hz, 3H), 2.97-2.94 (m, 4H), 1.60-1.53 (m, 4H), 1.31-1.19 (m, 36H), 0.78 (t, J=8.0 Hz, 6H) .sup.13C NMR (100 MHz, CD.sub.3OD): ? 163.7, 105.5, 78.2, 78.21, 75.3, 71.7, 70.3, 62.8, 61.5, 33.3, 31.2, 31.0, 30.7, 30.5, 30.3, 24.0, 14.8.

Preparation Example 17

Synthesis of TTG-A8

<17-1> Synthesis of Compound 3a

[0187] Compound 3a was synthesized in 80% yield according to Example 2-1. .sup.1H NMR (400 MHz, CDCl.sub.3): ? 5.86 (br, 2H, NH), 3.27-3.42 (m, 4H), 1.50-1.63 (m, 4H), 1.19-1.41 (m, 20H), 0.88 (t, J=7.8 Hz, 6H), .sup.13C NMR (100 MHz, CDCl.sub.3): 167.8, 65.4, 41.5, 32.5, 30.2, 29.8, 27.4, 23.3, 14.4.

<17-2> Synthesis of Compound 4a

[0188] Compound 4a was synthesized in 80% yield according to Example 2-2. .sup.1H NMR (400 MHz, CDCl.sub.3): ? 3.78-3.72 (m, 6H), 3.41-3.38 (m, 4H), 1.53-1.50 (m, 4H), 1.28-1.24 (m, 20H), 0.88 (t, J=7.2 Hz, 6H) .sup.13C NMR (100 MHz, CDCl.sub.3): ? 180.3, 179.4, 162.2, 62.4, 32.1, 30.1, 29.8, 29.4, 29.3, 29.1, 22.8, 14.2.

<17-3> Synthesis of TTG-A8a

[0189] TTG-A8a was synthesized in 85% yield according to the general glycosylation reaction procedure of Example 2-3. .sup.1H NMR (400 MHz, CDCl.sub.3): ? 8.08-8.01 (m, 12H), 7.91 (d, J=7.2 Hz, 6H), 7.83 (d, J=8.0 Hz, 6H), 7.71-7.26 (m, 46H), 5.97 (s, 1H), 5.60 (t, J=8.0 Hz, 3H), 5.44 (t, J=8.0 Hz 3H), 5.29-5.20 (m, 4H), 4.50-4.37 (m, 8H), 4.10-3.90 (m, 4H), 3.57-3.50 (m, 5H), 2.94-2.90 (m 4H), 1.59-1.55 (m, 4H), 1.26-1.21 (m, 20H), 0.88 (t, J=7.4 Hz, 6H), .sup.13C NMR (100 MHz, CDCl.sub.3): ? 166.2, 165.3, 165.1, 164.7, 162.6, 133.8, 133.6, 133.4, 133.3, 130.1, 129.9, 129.8, 129.6, 129.5, 129.2, 128.9, 128.8, 128.6, 128.4, 101.5, 72.6, 72.0, 71.8, 69.6, 68.3, 63.2, 59.8, 31.8, 29.2, 28.9, 22.7, 14.2.

<17-4> Synthesis of TTG-A8

[0190] TTG-A8 was synthesized in 85% yield according to the general synthesis procedure for the deprotection reaction of Example 2-4. .sup.1H NMR (400 MHz, CD.sub.3OD): ? 4.48 (d, J=8.0 Hz, 3H), 4.32-4.29 (m, 6H), 4.09-4.05 (m, 6H), 3.92-3.86 (m, 4H), 3.73-3.68 (m, 4H), 3.34-3.20 (m, 24H), 1.62-1.60 (m, 4H), 1.33-1.28 (m, 20H), 0.88 (t, J=7.8 Hz 6H) .sup.13C NMR (100 MHz, CD.sub.3OD): ? 164.7, 105.4, 78.2, 78.0, 75.1, 71.6, 70.2, 62.7, 33.2, 30.7, 30.6, 30.2, 23.6 14.5.

Preparation Example 18

Synthesis of TTG-A9

<18-1> Synthesis of Compound 3b

[0191] Compound 3b was synthesized in 78% yield according to Example 2-1. .sup.1H NMR (400 MHz, CDCl.sub.3): ? 5.87 (br, 2H, NH), 3.27-3.43 (m, 4H), 1.50-1.63 (m, 4H), 1.19-1.41 (m, 24H), 0.88 (t, J=7.8 Hz, 6H), .sup.13C NMR (100 MHz, CDCl.sub.3): 167.9, 65.5, 41.5, 32.5, 30.2, 29.9, 27.4, 23.3, 14.4.

<18-2> Synthesis of Compound 4b

[0192] Compound 4b was synthesized in 80% yield according to Example 2-2. .sup.1H NMR (400 MHz, CDCl.sub.3): ? 3.77-3.73 (m, 6H), 3.40-3.37 (m, 4H), 1.52-1.48 (m, 4H), 1.26-1.23 (m, 24H), 0.87 (t, J=7.2 Hz, 6H) .sup.13C NMR (100 MHz, CDCl.sub.3): ? 180.4, 179.5, 162.3, 62.5, 54.1, 32.1, 30.0, 29.8, 29.5, 29.4, 29.1, 22.8, 14.2.

<18-3> Synthesis of TTG-A9a

[0193] TTG-A9a was synthesized in 86% yield according to the general glycosylation reaction procedure of Example 2-3. .sup.1H NMR (400 MHz, CDCl.sub.3): ? 8.08-8.01 (m, 12H), 7.91 (d, J=7.2 Hz, 6H), 7.83 (d, J=8.0 Hz, 6H), 7.71-7.27 (m, 46H), 6.03 (s, 2H), 5.73-5.28 (m, 22H), 4.51-4.47 (m, 18H), 4.03-3.89 (m, 8H), 3.59-3.48 (m, 10H), 3.23-3.12 (m, 4H), 1.60-1.57 (m, 4H), 1.25-1.18 (m, 24H), 0.87 (t, J=7.4 Hz, 6H), .sup.13C NMR (100 MHz, CDCl.sub.3): ? 166.2, 165.1, 164.8, 162.5, 133.7, 133.5, 133.3, 133.2, 130.0, 129.8, 129.7, 129.6, 129.1, 128.8, 128.5, 128.4, 101.5, 101.4, 92.7, 72.6, 72.0, 71.8, 69.6, 68.2, 63.2, 59.8, 48.4, 31.9, 32.0, 29.7, 29.2, 29.0, 22.9, 14.2.

<18-4> Synthesis of TTG-A9

[0194] TTG-A9 was synthesized in 84% yield according to the general synthesis procedure for the deprotection reaction of Example 2-4. .sup.1H NMR (400 MHz, CD.sub.3OD): ? 4.48 (d, J=8.0 Hz, 3H), 4.31-4.29 (m, 6H), 4.09-4.06 (m, 6H), 3.91-3.86 (m, 4H), 3.73-3.67 (m, 4H), 3.34-3.21 (m, 24H), 1.62-1.60 (m, 4H), 1.33-1.28 (m, 24H), 0.88 (t, J=7.8 Hz, 6H) .sup.13C NMR (100 MHz, CD.sub.3OD): ? 164.6, 105.3, 78.2, 78.1, 75.1, 71.5, 70.2, 62.7, 33.2, 30.8, 30.5, 30.2, 23.5,14.5.

Preparation Example 19

Synthesis of TTG-A10

<19-1> Synthesis of Compound 3c

[0195] Compound 3c was synthesized in 82% yield according to Example 2-1. .sup.1H NMR (400 MHz, CDCl.sub.3): ? 5.86 (br, 2H, NH), 3.28-3.43 (m, 4H), 1.51-1.63 (m, 4H), 1.18-1.41 (m, 28H), 0.88 (t, J=7.8 Hz, 6H), .sup.13C NMR (100 MHz, CDCl.sub.3): 167.8, 165.6, 41.6, 32.5, 30.6, 29.9, 29.8, 27.4, 23.2, 14.3.

<19-2> Synthesis of Compound 4c

[0196] Compound 4c was synthesized in 80% yield according to Example 2-2. .sup.1H NMR (400 MHz, CDCl.sub.3): 6 3.78-3.74 (m, 6H), 3.41-3.38 (m, 4H), 1.53-1.48 (m, 4H), 1.27-1.24 (m, 28H), 0.88 (t, J=7.2 Hz, 6H) .sup.13C NMR (100 MHz, CDCl.sub.3): ? 180.2, 179.2, 162.2, 62.5, 54.2, 32.3, 30.0, 29.7, 29.3, 29.4, 29.0, 22.6, 14.2.

<19-3> Synthesis of TTG-A10a

[0197] TTG-A10a was synthesized in 86% yield according to the general glycosylation reaction procedure of Example 2-3. .sup.1H NMR (400 MHz, CDCl.sub.3): ? 8.08-8.01 (m, 12H), 7.91 (d, J=7.2 Hz, 6H), 7.83 (d, J=8.0 Hz, 6H), 7.71-7.27 (m, 46H), 6.03 (s, 2H), 5.73-5.28 (m, 22H), 4.51-4.47 (m, 18H), 4.03-3.89 (m, 8H), 3.59-3.48 (m, 10H), 3.23-3.12 (m, 4H), 1.60-1.57 (m, 4H), 1.25-1.18 (m, 28H), 0.87 (t, J=7.4 Hz, 6H), .sup.13C NMR (100 MHz, CDCl.sub.3): ? 166.2, 165.1, 164.7, 162.5, 133.8, 133.5, 133.3, 133.2, 130.0, 129.8, 129.7, 129.6, 129.1, 128.8, 128.5, 128.4, 101.5, 101.4, 92.7, 72.6, 72.0, 71.8, 69.6, 68.2, 63.2, 59.8, 48.4, 31.9, 32.0, 29.7, 29.2, 29.0, 22.9, 14.2.

<19-4> Synthesis of TTG-A10

[0198] TTG-A10 was synthesized in 86% yield according to the general synthesis procedure for the deprotection reaction of Example 2-4. .sup.1H NMR (400 MHz, CD.sub.3OD): ? 4.48 (d, J=8.0 Hz,3H), 4.32-4.29 (m, 6H), 4.08-4.05 (m, 6H), 3.91-3.87 (m, 4H), 3.72-3.67 (m, 4H), 3.33-3.21 (m, 24H), 1.62-1.60 (m, 4H), 1.33-1.28 (m, 28H), 0.88 (t, J=7.8 Hz, 6H) .sup.13C NMR (100 MHz, CD.sub.3OD): ? 164.8, 105.3, 78.2, 78.0, 75.1, 71.4, 70.3, 62.8, 33.3, 30.9, 30.2, 28.8, 23.7, 14.5.

Preparation Example 20

Synthesis of TTG-A11

<20-1> Synthesis of Compound 3d

[0199] Compound 3d was synthesized in 83% yield according to Example 2-1. .sup.1H NMR (400 MHz, CDCl.sub.3): ? 5.85 (br, 2H, NH), 3.28-3.41 (m, 4H), 1.52-1.63 (m, 4H), 1.18-1.42 (m, 32H), 0.88 (t, J=7.8 Hz, 6H), .sup.13C NMR (100 MHz, CDCl.sub.3): 167.8, 165.7, 41.5, 32.5, 30.6, 29.9, 29.7, 29.5, 27.4, 23.2, 14.3.

<20-2> Synthesis of Compound 4d

[0200] Compound 4d was synthesized in 84% yield according to Example 2-2. .sup.1H NMR (400 MHz, CDCl.sub.3): ? 3.77-3.73 (m, 6H), 3.41-3.38 (m, 4H), 1.53-1.549 (m, 4H), 1.25-1.23 (m, 32H), 0.88 (t, J=7.2 Hz, 6H) .sup.13C NMR (100 MHz, CDCl.sub.3): ? 180.3, 179.4, 162.2, 62.4, 54.1, 32.1, 30.1, 29.8, 29.5, 29.4, 29.1, 22.8, 14.2.

<20-3> Synthesis of TTG-A11a

[0201] TTG-A11a was synthesized in 84% yield according to the general glycosylation reaction procedure of Example 2-3. .sup.1H NMR (400 MHz, CDCl.sub.3): ? 8.08-8.01 (m, 12H), 7.91 (d, J=7.2 Hz, 6H), 7.83 (d, J=8.0 Hz, 6H), 7.71-7.27 (m, 46H), 6.03 (s, 2H), 5.73-5.28 (m, 22H), 4.51-4.47 (m, 18H), 4.03-3.89 (m, 8H), 3.59-3.48 (m, 10H), 3.23-3.12 (m, 4H), 1.60-1.57 (m, 4H), 1.25-1.18 (m, 32H), 0.87 (t, J=7.4 Hz, 6H), .sup.13C NMR (100 MHz, CDCl.sub.3): ? 166.2, 165.1, 164.7, 162.5, 133.8, 133.5, 133.3, 133.2, 130.0, 129.8, 129.7, 129.6, 129.1, 128.8, 128.5, 128.4, 101.5, 101.4, 92.7, 72.6, 72.0, 71.8, 69.6, 68.2, 63.2, 59.8, 48.4, 31.9, 32.0, 29.7, 29.2, 29.0, 22.9, 14.2.

<20-4> Synthesis of TTG-A11

[0202] TTG-A11 was synthesized in 85% yield according to the general synthesis procedure for the deprotection reaction of Example 2-4. .sup.1H NMR (400 MHz, CD.sub.3OD): ? 4.49 (d, J=8.0 Hz, 3H), 4.33-4.30 (m, 6H), 4.08-4.04 (m, 6H), 3.90-3.87 (m, 4H), 3.72-3.68 (m, 4H), 3.36-3.21 (m, 24H), 1.62-1.60 (m, 4H), 1.32-1.28 (m, 32H), 0.88 (t, J=7.8 Hz 6H) .sup.13C NMR (100 MHz, CD.sub.3OD): ? 164.7, 105.4, 78.1, 78.0, 75.1, 71.5, 70.3, 62.7, 33.2, 30.9, 30.6, 30.3, 28.8, 23.8, 14.5.

Preparation Example 21

Synthesis of TTG-A12

<21-1> Synthesis of Compound 3e

[0203] Compound 3e was synthesized in 85% yield according to Example 2-1. .sup.1H NMR (400 MHz, CDCl.sub.3): ? 5.84 (br, 2H, NH), 3.29-3.41 (m, 4H), 1.53-1.63 (m, 4H), 1.18-1.42 (m, 36H), 0.88 (t, J=7.8 Hz, 6H), .sup.13C NMR (100 MHz, CDCl.sub.3): 167.8, 165.5, 41.5, 32.7, 30.6, 29.9, 29.7, 29.5, 27.8, 23.1, 14.2.

<21-2> Synthesis of Compound 4e

[0204] Compound 4e was synthesized in 84% yield according to Example 2-2. .sup.1H NMR (400 MHz, CDCl.sub.3): ? 3.77-3.73 (m, 6H), 3.41-3.38 (m, 4H), 1.53-1.549 (m, 4H), 1.25-1.23 (m, 32H), 0.88 (t, J=7.2 Hz, 6H) .sup.13C NMR (100 MHz, CDCl.sub.3): ? 180.3, 179.4, 162.2, 62.4, 54.1, 32.1, 30.1, 29.8, 29.5, 29.4, 29.1, 22.8, 14.2.

<21-3> Synthesis of TTG-A12a

[0205] TTG-A12a was synthesized in 85% yield according to the general glycosylation reaction procedure of Example 2-3. .sup.1H NMR (400 MHz, CDCl.sub.3): ? 8.08-8.01 (m, 12H), 7.91 (d, J=7.2 Hz, 6H), 7.83 (d, J=8.0 Hz, 6H), 7.71-7.27 (m, 46H), 6.04 (s, 2H), 5.71-5.26 (m, 22H), 4.51-4.47 (m, 18H), 4.03-3.89 (m, 8H), 3.59-3.48 (m, 10H), 3.23-3.12 (m, 4H), 1.59-1.57 (m, 4H), 1.26-1.18 (m, 36H), 0.87 (t, J=7.4 Hz, 6H), .sup.13C NMR (100 MHz, CDCl.sub.3): ? 166.1, 165.3, 164.5, 162.2, 133.8, 133.5, 133.3, 133.2, 130.0, 129.8, 129.7, 129.6, 129.1, 128.8, 128.5, 128.4, 101.5, 101.4, 92.7, 72.6, 72.0, 71.8, 69.6, 68.2, 63.1, 59.8, 48.4, 31.9, 32.0, 29.7, 29.3, 29.0, 22.8, 14.2.

<21-4> Synthesis of TTG-A12

[0206] TTG-A12 was synthesized in 84% yield according to the general synthesis procedure for the deprotection reaction of Example 2-4. .sup.1H NMR (400 MHz, CD.sub.3OD): ? 4.49 (d, J=8.0 Hz, 3H), 4.33-4.29 (m, 6H), 4.08-4.05 (m, 6H), 3.89-3.86 (m, 4H), 3.71-3.68 (m, 4H), 3.36-3.21 (m, 24H), 1.63-1.60 (m, 4H), 1.32-1.28 (m, 36H), 0.89 (t, J=7.8 Hz, 6H) .sup.13C NMR (100 MHz, CD.sub.3OD): ? 164.8, 105.4, 78.1, 78.0, 75.1, 71.5, 70.3, 62.7, 33.2, 30.9, 30.7, 30.6, 30.3, 23.8, 23.5,14.5.

Preparation Example 22

Synthesis of TTG-A14

<22-1> Synthesis of Compound 3f

[0207] Compound 3f was synthesized in 84% yield according to Example 2-1. .sup.1H NMR (400 MHz, CDCl.sub.3): ? 5.85 (br, 2H, NH), 3.29-3.41 (m, 4H), 1.53-1.63 (m, 4H), 1.18-1.42 (m, 44H), 0.88 (t, J=7.2 Hz, 6H), .sup.13C NMR (100 MHz, CDCl.sub.3): 167.8, 165.5, 41.5, 32.7, 31.8, 30.6, 29.9, 29.7, 29.5, 27.8, 23.1, 14.2.

<22-2> Synthesis of Compound 4f

[0208] Compound 4f was synthesized in 86% yield according to Example 2-2. .sup.1H NMR (400 MHz, CDCl.sub.3): ? 3.78-3.74 (m, 6H), 3.42-3.28 (m, 4H), 1.55-1.39(m, 4H), 1.29-1.25 (m, 44H), 0.87 (t, J=7.2 Hz, 6H) .sup.13C NMR (100 MHz, CDCl.sub.3): ? 180.4, 179.2, 162.4, 62.5, 54.2, 32.1, 30.0, 29.9, 29.7, 29.6, 29.5, 29.4, 29.2, 22.6, 14.2.

<22-3> Synthesis of TTG-A14a

[0209] TTG-A14a was synthesized in 86% yield according to the general glycosylation reaction procedure of Example 2-3. .sup.1H NMR (400 MHz, CDCl.sub.3): ? 8.08-8.01 (m, 12H), 7.91 (d, J=7.2 Hz, 6H), 7.83 (d, J=8.0 Hz, 6H), 7.71-7.27 (m, 46H), 6.03 (s, 2H), 5.73-5.28 (m, 22H), 4.51-4.47 (m, 18H), 4.03-3.89 (m, 8H), 3.59-3.48 (m, 10H), 3.23-3.12 (m, 4H), 1.60-1.57 (m, 4H), 1.25-1.18 (m, 44H), 0.87 (t, J=7.4 Hz, 6H), .sup.13C NMR (100 MHz, CDCl.sub.3): ? 166.1, 165.1, 164.7, 162.5, 133.8, 133.5, 133.3, 133.2, 130.0, 129.8, 129.7, 129.6, 129.1, 128.8, 128.5, 128.4, 101.5, 101.4, 92.7, 72.6, 72.0, 71.8, 69.6, 68.2, 63.2, 59.8, 48.4, 31.9, 31.6, 32.0, 29.7, 29.2, 29.0, 22.9, 14.2.

<22-4> Synthesis of TTG-A14

[0210] TTG-A14 was synthesized in 86% yield according to the general synthesis procedure for the deprotection reaction of Example 2-4. .sup.1H NMR (400 MHz, CD.sub.3OD): ? 4.48 (d, J=8.0 Hz, 3H), 4.34-4.30 (m, 6H), 4.07-4.05 (m, 6H), 3.88-3.85 (m, 4H), 3.72-3.68 (m, 4H), 3.35-3.21 (m, 24H), 1.63-1.60 (m, 4H), 1.31-1.28 (m, 44H), 0.88 (t, J=7.8 Hz, 6H) .sup.13C NMR (100 MHz, CD.sub.3OD): ? 164.7, 105.3, 78.1, 78.2, 75.8, 71.4, 70.3, 62.6, 33.2, 30.8, 30.5, 30.2, 23.8, 23.3 14.5.

EXAMPLE 3

Synthesis Method of TEMs

[0211] FIG. 3 illustrates the synthesis scheme of triazine-based maltosides with a diethanolamine linker (TEMs). 10 types of compounds of TEMs were synthesized by the following synthesis methods of <3-1> to <3-4>.

<3-1> General Procedure for the Synthesis of 2-chloro-4,6-dialkylated-1,3,5-triazine (Step i of FIG. 3)

[0212] A mixture of 2,4,6-trichloro-1,3,5-triazine (3.01 mmol) and NaHCO.sub.3 (7.26 mmol) was stirred in acetone (10 mL) for 10 minutes. Each alcohol (ROH/RSH) (6.0 mmol) dissolved in acetone was added dropwise for 30 minutes. The resulting reaction mixture was kept at room temperature for 36 hr for ROH or 1 hr for RSH. The reaction mixture was extracted with CHCl.sub.3 and water, and the organic layer was dried over anhydrous Na.sub.2SO.sub.4. The oily residue obtained after removal of solvent was subjected to column chromatography purification to obtain target Compound 1 or 3.

<3-2> General Synthesis Procedure for the Coupling Reactions of the Resulting Dialkylated Triazine Derivatives With Diethanolamine (Step ii of FIG. 3)

[0213] To a mixture of 2-chloro-4,6-dialkylated-1,3,5-triazine (1.0 equiv.) dissolved in THF, diethanolamine and K.sub.2CO.sub.3 were added under nitrogen. The solution was stirred at 40? C. for 24 hr. The reaction mixture was diluted with water and then extracted with ethyl acetate. The organic layer was washed with brine and dried over anhydrous Na.sub.2SO.sub.4. After evaporation of the ethyl acetate solution, the residue was purified by flash column chromatography (EtOAc/hexane) to obtain target Compound 5 or 6.

<3-3> General Synthesis Procedure for Glycosylation Reaction (Step iii of FIG. 3)

[0214] This reaction was performed according to the synthesis method (Nat. Methods 2010, 7, 1003.) of Chae, P. S. et al. with some modifications. Briefly, a mixture of a dialkylated di-ol derivative (Compound 5 or 6), AgOTf (2.5 equiv.), 2,4,6-collidine (0.5 equiv.) in anhydrous CH.sub.2Cl.sub.2 (20 mL) was stirred at ?45? C. Then, perbenzoylated maltosylbromide (2.5 equiv.) dissolved in CH.sub.2Cl.sub.2 (30 mL) was added dropwise over 0.5 hr into this suspension. The reaction was maintained at 0? C. for 1.5 hr. Reaction progress was monitored by TLC. After completion of the reaction (as detected by TLC), pyridine (1.0 mL) was added to the reaction mixture. The reaction mixture was diluted with CH.sub.2Cl.sub.2 (30 mL) before being filtered over Celite. The filtrate was washed successively with a 1.0 M aqueous Na.sub.2S.sub.2O.sub.3 solution (30 mL), a 0.1 M aqueous HCl solution (30 mL), and brine (30 mL). Then, the organic layer was dried with anhydrous Na.sub.2SO.sub.4 and the solvent was removed by rotary evaporation. The resulting residue was purified by silica gel column chromatography (EtOAc/hexane) to obtain the glycosylated target compound.

<3-4> General Procedure for the Synthesis For Deprotection Reaction (Step iv of FIG. 3)

[0215] This reaction followed the synthesis method (Nat. Methods 2010, 7, 1003.) of Chae, P. S. aureus et al. The de-O-benzoylation was performed under Zemplen's condition. An O-protected compound was dissolved in anhydrous CH.sub.2Cl.sub.2 and then MeOH was added slowly thereto until persistent precipitation appeared. A methanolic solution of 0.5 M NaOMe was added to the reaction mixture such that the final concentration of NaOMe was 0.05 M. The reaction mixture was stirred for 6 hr at room temperature. After completion of the reaction, the reaction mixture was neutralized using Amberlite IR-120 (H.sup.+ form) resin. The resin was removed by filtration and washed with MeOH and the solvent was removed from the filtrate in vacuo. The residue was purified by silica gel column chromatography (CH.sub.2Cl.sub.2/MeOH) to obtain the target compound.

Preparation Example 23

Synthesis of TEM-E7

<23-1> Synthesis of Compound 1a

[0216] Compound 1a was synthesized in 48% yield according to Example 3-1. .sup.1H NMR (400 MHz, CDCl.sub.3): ? 4.41 (t, J=6.6 Hz, 4H), 1.82-1.75 (m, 4H), 1.34-1.27 (m, 16H), 0.88 (t, J=6.9 Hz, 6H); .sup.13C NMR (100 MHz, CDCl.sub.3): ? 172.7, 172.2, 69.6, 32.0, 29.6, 28.6, 25.9, 22.8, 14.2.

<23-2> Synthesis of Compound 5a

[0217] Compound 5a was synthesized in 85% yield according to Example 3-2. .sup.11H NMR (400 MHz, CDCl.sub.3): ? 4.29 (t, J=6.7 Hz, 4H), 3.92 (t, J=7.0 Hz, 4H), 3.80 (t, J=7.0 Hz, 4H), 1.77-1.72 (m, 4H), 1.39-1.26 (m, 16H), 0.88 (t, J=7.2 Hz, 6H); .sup.13C NMR (100 MHz, CDCl.sub.3): ? 171.6, 168.2, 67.9, 62.0, 52.4, 32.0, 29.6, 29.4, 28.9, 26.6, 22.8, 14.2.

<23-3> Synthesis of TEM-E7a

[0218] TEM-E7a was synthesized in 82% yield according to the general glycosylation reaction procedure of Example 3-3. .sup.1H NMR (400 MHz, CDCl.sub.3): ? 8.11 (d, J=7.1 Hz, 4H), 8.01 (d, J=7.2 Hz, 4H), 7.82 (d, J=7.2 Hz, 4H), 7.79-7.73 (m, 12H), 7.61 (d, J=7.4 Hz, 4H), 7.52-7.12 (m, 42H), 6.11 (t, J=10.0 Hz, 2H), 5.76-5.62 (m, 6H), 5.45-5.2130 (m, 4H), 4.92 (d, J=10.4 Hz, 2H), 4.78-4.75 (m, 2H), 4.65-4.56 (m, 4H), 4.47-4.24 (m, 4H), 4.10-4.03 (m, 6H), 3.81-3.78 (m, 2H), 3.58-3.49 (m, 4H), 1.67-1.62 (m, 4H), 1.32-1.22 (m, 20H), 0.88 (t, J=7.0 Hz, 6H); .sup.13C NMR (100 MHz, CDCl.sub.3): ? 172.3, 171.3, 166.5, 166.2, 165.8, 165.7, 165.4, 165.1, 161.0, 164.9, 133.5, 133.4, 133.3, 133.1, 130.0, 129.9, 129.8, 129.7, 129.5, 129.3, 129.0, 128.9, 128.6, 128.4, 128.3, 128.2, 128.1, 100.8, 96.3, 74.9, 72.8, 72.2, 70.8, 69.9, 69.1, 67.3, 63.4, 62.5, 48.8, 31.8, 29.5, 29.4, 29.3, 29.0, 28.7, 25.9, 22.6, 14.1.

<23-4> Synthesis of TEM-E7

[0219] TEM-E7 was synthesized in 95% yield according to the general synthesis procedure for the deprotection reaction of Example 3-4. .sup.1H NMR (400 MHz, CD.sub.3OD): ? 5.16 (d, J=7.0 Hz, 2H), 4.39-4.35 (m, 6H), 4.04-3.97 (m, 2H), 3.83-3.71 (m, 12H), 3.58-3.42 (m, 10H), 3.37-3.28 (m, 4H), 1.66-1.59 (m, 4H), 1.32-1.18 (m, 16H), 0.78 (t, J=7.2 Hz, 6H); .sup.13C NMR (100 MHz, CD.sub.3OD): ? 173.2, 172.8, 169.0, 104.9, 104.7, 103.0, 81.3, 81.2, 77.7, 76.7, 75.1, 74.8, 74.7, 74.2, 71.5, 68.9, 68.7, 62.8, 62.2, 32.0, 33.0, 30.7, 30.5, 30.2, 30.0, 27.1, 27.0, 23.9, 23.7, 14.6, 14.5; HRMS (FAB.sup.+): For C.sub.45H.sub.80N.sub.4O.sub.24 [M+Na].sup.+ 1083.5060, found 1083.5058.

Preparation Example 24

Synthesis of TEM-E8

<24-1> Synthesis of Compound 1b

[0220] Compound 1b was synthesized in 45% yield according to Example 3-1. .sup.1H NMR (400 MHz, CDCl.sub.3): ? 4.41 (t, J=6.6 Hz, 4H), 1.81-1.72 (m, 4H), 1.34-1.27 (m, 20H), 0.86 (t, J=6.9 Hz, 6H); .sup.13C NMR (100 MHz, CDCl.sub.3): ? 172.7, 172.2, 69.6, 32.0, 29.7, 29.6, 28.6, 25.9, 22.8, 14.2.

<24-2> Synthesis of Compound 5b

[0221] Compound 5b was synthesized in 88% yield according to Example 3-2. .sup.1H NMR (400 MHz, CDCl.sub.3): ? 4.29 (t, J=6.7 Hz, 4H), 3.90 (t, J=7.1 Hz, 4H), 3.79 (t, J=7.0 Hz, 4H), 1.76-1.73 (m, 4H), 1.40-1.27 (m, 20H), 0.88 (t, J=7.2 Hz, 3H), 0.87 (t, J=7.2 Hz, 3H); .sup.13C NMR (100 MHz, CDCl.sub.3): ? 171.5, 167.8, 67.8, 61.6, 52.6, 31.9, 29.4, 29.3, 28.8, 26.0, 22.7, 14.1.

<24-3> Synthesis of TEM-E8a

[0222] TEM-E8a was synthesized in 82% yield according to the general glycosylation reaction procedure of Example 3-3. .sup.1H NMR (400 MHz, CDCl.sub.3): ? 8.11 (d, J=7.1 Hz, 4H), 8.01 (d, J=7.2 Hz, 4H), 7.82 (d, J=7.2 Hz, 4H), 7.79-7.73 (m, 12H), 7.61 (d, J=7.4 Hz, 4H), 7.52-7.12 (m, 42H), 6.11 (t, J=10.0 Hz, 2H), 5.76-5.62 (m, 6H), 5.45-5.2130 (m, 4H), 4.92 (d, J=10.4 Hz, 2H), 4.78-4.75 (m, 2H), 4.65-4.56 (m, 4H), 4.47-4.24 (m, 4H), 4.10-4.03 (m, 6H), 3.81-3.78 (m, 2H), 3.58-3.49 (m, 4H), 1.67-1.62 (m, 4H), 1.32-1.22 (m, 20H), 0.88 (t, J=7.0 Hz, 6H); .sup.13C NMR (100 MHz, CDCl.sub.3): ? 172.3, 171.3, 166.5, 166.2, 165.8, 165.7, 165.4, 165.1, 161.0, 164.9, 133.5, 133.4, 133.3, 133.1, 130.0, 129.9, 129.8, 129.7, 129.5, 129.3, 129.0, 128.9, 128.6, 128.4, 128.3, 128.2, 128.1, 100.8, 96.3, 74.9, 72.8, 72.2, 70.8, 69.9, 69.1, 67.3, 63.4, 62.5, 48.8, 31.8, 29.5, 29.4, 29.3, 29.0, 28.7, 25.9, 22.6, 14.1.

<24-4> Synthesis of TEM-E8

[0223] TEM-E8 was synthesized in 90% yield according to the general synthesis procedure for the deprotection reaction of Example 3-4. .sup.1H NMR (400 MHz, CD.sub.3OD): ? 5.17 (d, J=7.0 Hz, 2H), 4.39-4.35 (m, 6H), 4.04-3.97 (m, 2H), 3.83-3.71 (m, 12H), 3.58-3.42 (m, 10H), 3.37-3.28 (m, 4H), 1.66-1.59 (m, 4H), 1.32-1.18 (m, 20H), 0.78 (t, J=7.2 Hz, 6H); .sup.13C NMR (100 MHz, CD.sub.3OD): ? 172.6, 168.2, 104.6, 102.9, 81.2, 77.9, 76.6, 75.1, 74.8, 74.7, 74.2, 71.5, 69.0, 68.7, 62.8, 62.2, 50.0, 49.5, 49.3, 49.1, 48.9, 48.7, 33.0, 30.5, 30.4, 30.0, 27.1, 23.8, 14.6; HRMS (FAB.sup.+): For C.sub.47H.sub.84N.sub.4O.sub.24 [M+Na].sup.+ 1111.5373, found 1111.5377.

Preparation Example 25

Synthesis of TEM-E9

<25-1> Synthesis of Compound 1c

[0224] Compound 1c was synthesized in 50% yield according to Example 3-1. .sup.1H NMR (400 MHz, CDCl.sub.3): ? 4.41 (t, J=6.6 Hz, 4H), 1.82-1.72 (m, 4H), 1.34-1.27 (m, 24H), 0.88 (t, J=6.9 Hz, 6H); .sup.13C NMR (100 MHz, CDCl.sub.3): ? 172.7, 172.2, 69.6, 32.0, 29.6, 29.4, 28.6, 25.9, 22.8, 14.2.

<25-2> Synthesis of Compound 5c

[0225] Compound 5c was synthesized in 84% yield according to Example 3-2. .sup.1H NMR (400 MHz, CDCl.sub.3): ? 4.28 (t, J=6.7 Hz, 4H), 3.90 (t, J=7.3 Hz, 4H), 3.79 (t, J=7.3 Hz, 4H), 1.77-1.72 (m, 4H), 1.39-1.26 (m, 24H), 0.87 (t, J=6.9 Hz, 6H); .sup.13C NMR (100 MHz, CDCl.sub.3): ? 171.6, 168.2, 67.9, 62.0, 52.4, 32.0, 29.6, 29.4, 28.9, 26.6, 22.8, 14.2.

<25-3> Synthesis of TEM-E9a

[0226] TEM-E9a was synthesized in 85% yield according to the general glycosylation reaction procedure of Example 3-3. .sup.1H NMR (400 MHz, CDCl.sub.3): ? 8.11 (d, J=7.1 Hz, 4H), 8.01 (d, J=7.2 Hz, 4H), 7.82 (d, J=7.2 Hz, 4H), 7.79-7.73 (m, 12H), 7.61 (d, J=7.4 Hz, 4H), 7.52-7.12 (m, 42H), 6.11 (t, J=8.0 Hz, 2H), 5.76-5.62 (m, 6H), 5.45-5.2130 (m, 4H), 4.92 (d, J=8.2 Hz, 2H), 4.78-4.75 (m, 2H), 4.65-4.56 (m, 4H), 4.47-4.24 (m, 6H), 4.10-4.03 (m, 6H), 3.81-3.78 (m, 2H), 3.58-3.49 (m, 4H), 1.67-1.62 (m, 4H), 1.32-1.22 (m, 24H), 0.88 (t, J=7.0 Hz, 6H); .sup.13C NMR (100 MHz, CDCl.sub.3): ? 172.3, 171.01, 166.5, 166.2, 165.8, 165.7, 165.4, 165.1, 161.0, 164.9, 133.5, 133.4, 133.3, 133.1, 130.0, 129.9, 129.8, 129.7, 129.5, 129.3, 129.0, 128.9, 128.6, 128.4, 128.3, 128.2, 128.1, 100.8, 96.3, 74.9, 72.8, 72.2, 70.8, 69.9, 69.1, 68.6, 67.3, 63.4, 62.5, 48.8, 31.9, 31.6, 29.5, 29.4, 29.3, 29.0, 28.7, 25.9, 22.6, 14.2.

<25-4> Synthesis of TEM-E9

[0227] TEM-E9 was synthesized in 90% yield according to the general synthesis procedure for the deprotection reaction of Example 3-4. .sup.1H NMR (400 MHz, CD.sub.3OD): ? 5.18 (d, J=7.0 Hz, 2H), 4.35-4.30 (m, 6H), 4.04-3.97 (m, 2H), 3.83-3.71 (m, 12H), 3.58-3.42 (m, 10H), 3.37-3.28 (m, 4H), 1.66-1.59 (m, 4H), 1.32-1.18 (m, 24H), 0.78 (t, J=7.2 Hz, 6H); .sup.13C NMR (100 MHz, CD.sub.3OD): ? 172.6, 168.2, 104.6, 102.9, 81.2, 77.6, 75.1, 74.8, 74.6, 74.1, 71.4, 69.0, 68.7, 62.7, 50.0, 49.7, 49.5, 49.3, 48.9, 48.7, 48.5, 33.1, 30.7, 30.5, 30.4, 30.0, 27.1, 23.8, 14.6; HRMS (FAB.sup.+): For C.sub.49H.sub.88N.sub.4O.sub.24 [M+Na].sup.+ 1139.5686, found 1139.5682.

Preparation Example 26

Synthesis of TEM-E10

<26-1> Synthesis of Compound 1d

[0228] Compound 3d was synthesized in 50% yield according to Example 3-1. .sup.1H NMR (400 MHz, CDCl.sub.3): ? 4.41 (t, J=6.6 Hz, 4H), 1.81-1.77 (m, 4H), 1.34-1.27 (m, 28H), 0.88 (t, J=6.9 Hz, 6H); .sup.13C NMR (100 MHz, CDCl.sub.3): ? 172.7, 172.2, 69.6, 64.3, 32.0, 29.7, 29.6, 29.4, 28.6, 25.9, 22.8, 14.2.

<26-2> Synthesis of Compound 5d

[0229] Compound 5d was synthesized in 85% yield according to Example 3-2. .sup.1H NMR (400 MHz, CDCl.sub.3): ? 4.28 (t, J=6.6 Hz, 4H), 3.87 (t, J=7.3 Hz, 4H), 3.74 (t, J=7.3 Hz, 4H), 1.77-1.72 (m, 4H), 1.39-1.26 (m, 28H), 0.87 (t, J=6.9 Hz, 6H); .sup.13C NMR (100 MHz, CDCl.sub.3): ? 171.6, 167.2, 67.8, 61.7, 52.6, 32.0, 29.6, 29.4, 28.9, 26.6, 22.8, 14.2.

<26-3> Synthesis of TEM-E10a

[0230] TEM-E10a was synthesized in 80% yield according to the general glycosylation reaction procedure of Example 3-3. .sup.1H NMR (400 MHz, CDCl.sub.3): ? 8.11 (d, J=7.1 Hz, 4H), 8.01 (d, J=7.2 Hz, 4H), 7.82 (d, J=7.2 Hz, 4H), 7.79-7.73 (m, 12H), 7.61 (d, J=7.4 Hz, 4H), 7.52-7.12 (m, 42H), 6.11 (t, J=10.0 Hz, 2H), 5.76-5.62 (m, 6H), 5.45-5.21 (m, 4H), 4.92 (d, J=8.2 Hz, 2H), 4.78-4.75 (m, 2H), 4.65-4.56 (m, 4H), 4.47-4.24 (m, 6H), 4.10-4.03 (m, 6H), 3.81-3.78 (m, 2H), 3.58-3.49 (m, 4H), 1.67-1.62 (m, 4H), 1.32-1.22 (m, 28H), 0.88 (t, J=7.0 Hz, 6H); 13 C NMR (100 MHz, CDCl.sub.3): ? 172.3, 171.0, 166.5, 166.2, 165.8, 165.7, 165.4, 165.1, 161.0, 164.9, 133.5, 133.4, 133.3, 133.1, 130.0, 129.9, 129.8, 129.7, 129.5, 129.3, 129.0, 128.9, 128.6, 128.4, 128.3, 128.2, 128.1, 100.8, 96.3, 74.9, 72.8, 72.2, 70.8, 69.9, 69.1, 68.6, 67.6, 63.4, 62.5, 48.8, 31.9, 29.6, 29.4, 29.3, 29.0, 28.7, 25.9, 22.7, 14.2.

<26-4> Synthesis of TEM-E10

[0231] TEM-E10 was synthesized in 95% yield according to the general synthesis procedure for the deprotection reaction of Example 3-4. .sup.1H NMR (400 MHz, CD.sub.3OD): 6 5.17 (d, J=7.0 Hz, 2H), 4.38-4.35 (m, 6H), 3.99-3.96 (m, 2H), 3.82-3.70 (m, 12H), 3.57-3.36 (m, 10H), 3.27-3.14 (m, 4H), 1.63-1.60 (m, 4H), 1.31-1.17 (m, 28H), 0.78 (t, J=7.2 Hz, 6H); .sup.13C NMR (100 MHz, CD.sub.3OD): ? 172.6, 168.2, 104.6, 102.9, 81.2, 77.8, 76.6, 75.1, 74.8, 74.6, 74.1, 71.4, 69.0, 68.7, 62.7, 62.2, 50.0, 49.7, 49.5, 49.3, 49.1, 48.9, 48.7, 48.5, 33.1, 30.7, 30.5, 30.0, 27.1, 23.8, 14.6; HRMS (FAB.sup.+): For C.sub.51H.sub.92N.sub.4O.sub.24 [M+Na].sup.+ 1167.5999, found 1167.6001.

Preparation Example 27

Synthesis of TEM-E11

<27-1> Synthesis of Compound 1e

[0232] Compound 1e was synthesized in 48% yield according to Example 3-1. .sup.1H NMR (400 MHz, CDCl.sub.3): ? 4.41 (t, J=6.6 Hz, 4H), 1.82-1.72 (m, 4H), 1.34-1.27 (m, 32H), 0.88 (t, J=6.9 Hz, 6H); .sup.13C NMR (100 MHz, CDCl.sub.3): ? 172.7, 172.2, 69.6, 32.0, 29.7, 29.6, 29.6, 29.3, 28.5, 25.8, 22.8, 14.2.

<27-2> Synthesis of Compound 5e

[0233] Compound 5e was synthesized in 85% yield according to Example 3-2. .sup.1H NMR (400 MHz, CDCl.sub.3): ? 4.28 (m, J=6.6 Hz, 4H), 3.90 (m, 4H), 3.79 (m, 4H), 1.76-1.72 (m, 4H), 1.39-1.26 (m, 32H), 0.88 (t, J=7.2 Hz, 3H), 0.87 (t, J=7.2 Hz, 3H); .sup.13C NMR (100 MHz, CDCl.sub.3): ? 171.5, 167.8, 67.8, 61.60, 52.6, 32.0, 29.7, 29.6, 29.4, 28.8, 26.0, 22.8, 14.2.

<27-3> Synthesis of TEM-E11a

[0234] TEM-E11a was synthesized in 82% yield according to the general glycosylation reaction procedure of Example 3-3. .sup.1H NMR (400 MHz, CDCl.sub.3): ? 8.11 (d, J=7.1 Hz, 4H), 8.01 (d, J=7.2 Hz, 4H), 7.82 (d, J=7.2 Hz, 4H), 7.79-7.73 (m, 12H), 7.61 (d, J=7.4 Hz, 4H), 7.52-7.12 (m, 42H), 6.11 (t, J=10.0 Hz, 2H), 5.76-5.62 (m, 6H), 5.45-5.2130 (m, 4H), 4.92 (d, J=10.4 Hz, 2H), 4.78-4.75 (m, 2H), 4.65-4.56 (m, 4H), 4.47-4.24 (m, 6H), 4.10-4.03 (m, 6H), 3.81-3.78 (m, 2H), 3.58-3.49 (m, 4H), 1.67-1.62 (m, 4H), 1.32-1.22 (m, 32H), 0.88 (t, J=7.0 Hz, 6H); .sup.13C NMR (100 MHz, CDCl.sub.3): ? 172.3, 171.4, 166.5, 166.2, 165.8, 165.7, 165.4, 165.1, 161.0, 164.9, 133.5, 133.4, 133.3, 133.1, 130.0, 129.9, 129.8, 129.7, 129.5, 129.3, 129.0, 128.9, 128.6, 128.4, 128.3, 128.2, 128.1, 100.8, 96.3, 75.01, 72.9, 72.2, 70.9, 70.2, 69.2, 68.7, 67.4, 63.5, 62.5, 48.8, 32.0, 29.7, 29.7, 29.5, 29.4, 28.8, 26.0, 22.8, 14.2.

<27-4> Synthesis of TEM-E11

[0235] TEM-E11 was synthesized in 90% yield according to the general synthesis procedure for the deprotection reaction of Example 3-4. .sup.1H NMR (400 MHz, CD.sub.3OD): ? 5.16 (d, J=7.0 Hz, 2H), 4.39-4.34 (m, 6H), 3.99-3.97 (m, 2H), 3.83-3.57 (m, 12H), 3.41-3.26 (m, 10H), 3.20-3.13 (m, 4H), 1.66-1.61 (m, 4H), 1.32-1.18 (m, 32H), 0.79 (t, J=7.2 Hz, 6H); .sup.13C NMR (100 MHz, CD.sub.3OD): ? 172.7, 168.3, 104.7, 103.0, 81.3, 77.9, 76.7, 75.1, 74.8, 74.7, 74.2, 71.5, 69.0, 68.7, 62.8, 62.2, 50.0, 49.7, 49.5, 49.3, 49.1, 48.9, 48.7, 48.5, 33.2, 30.8, 30.6, 30.0, 27.1, 23.8, 14.6; HRMS (FAB.sup.+): C.sub.53H.sub.96N.sub.4O.sub.24 [M+Na].sup.+ 1195.6312, found 1195.6306.

Preparation Example 28

Synthesis of TEM-T7

<28-1> Synthesis of Compound 3a

[0236] Compound 3a was synthesized in 85% yield according to Example 3-1. .sup.1H NMR (400 MHz, CDCl.sub.3): ? 4.42 (t, J=6.6 Hz, 4H), 1.81-1.77 (m, 4H), 1.44-1.30 (m, 16H), 0.89 (t, J=6.9 Hz, 6H); .sup.13C NMR (100 MHz, CDCl.sub.3): ? 172.7, 172.2, 69.6, 32.0, 29.6, 28.6, 25.9, 22.8, 14.2.

<28-2> Synthesis of Compound 6a

[0237] Compound 6a was synthesized in 82% yield according to Example 3-2. .sup.1H NMR (400 MHz, CDCl.sub.3): ? 4.65 (bs, 2H), 3.90-3.88 (m, 4H), 3.78-3.75 (m, 4H), 2.99 (t, J=7.4 Hz, 4H), 1.71-1.67 (m, 4H), 1.42-1.27 (m, 16H), 0.88 (t, J=7.0 Hz, 6H); .sup.13C NMR (100 MHz, CDCl.sub.3): ? 179.2, 162.1, 61.3, 52.8, 31.8, 30.1, 29.5, 29.0, 28.9, 22.6, 14.1.

<28-3> Synthesis of TEM-T7a

[0238] TEM-T7a was synthesized in 85% yield according to the general glycosylation reaction procedure of Example 3-3. .sup.1H NMR (400 MHz, CDCl.sub.3): ? 8.12 (d, J=7.2 Hz, 4H), 8.00 (d, J=7.2 Hz, 4H), 7.82 (d, J=7.2 Hz, 4H), 7.79-7.73 (m, 12H), 7.61 (d, J=7.3 Hz, 4H), 7.56-7.12 (m, 42H), 6.11 (t, J=10.0 Hz, 2H), 5.77-5.66 (m, 4H), 5.41-5.21 (m, 4H), 4.90 (d, J=10.5 Hz, 2H), 4.79-4.75 (m, 2H), 4.62-4.46 (m, 4H), 4.47-4.23 (m, 4H), 4.12-4.03 (m, 4H), 3.76 (t, J=7.0 Hz, 2H), 3.54-3.45 (m, 4H), 2.78 (t, J=7.2 Hz, 2H) 1.57-1.53 (m, 4H), 1.29-1.20 (m, 16H), 0.88 (t, J=7.0 Hz, 6H); .sup.13C NMR (100 MHz, CDCl.sub.3): ? 172.3, 171.3, 166.5, 166.2, 165.8, 165.7, 165.4, 165.1, 161.0, 164.9, 133.5, 133.4, 133.3, 133.1, 130.0, 129.9, 129.8, 129.7, 129.5, 129.3, 129.0, 128.9, 128.6, 128.4, 128.3, 128.2, 128.1, 100.8, 96.3, 74.9, 72.8, 72.2, 70.8, 69.9, 69.1, 67.3, 63.4, 62.5, 48.8, 48.8, 31.8, 29.5, 29.3, 29.0, 28.7, 25.9, 22.6, 14.1.

<28-4> Synthesis of TEM-T7

[0239] TEM-T7 was synthesized in 90% yield according to the general synthesis procedure for the deprotection reaction of Example 3-4. .sup.1H NMR (400 MHz, CD.sub.3OD): ? 5.16 (d, J=7.0 Hz, 2H), 4.33 (d, J=8.0 Hz, 2H), 3.96-3.92 (m, 2H), 3.83-3.79 (m, 4H), 3.75-3.65 (m, 8H), 3.54-3.38 (m, 10H), 3.32-3.22 (m, 4H), 3.15-3.10 (m, 4H), 2.91 (t, J=7.2 Hz, 4H), 1.60-1.53 (m, 4H), 1.31-1.16 (m, 16H), 0.78 (t, J=7.0 Hz, 6H); .sup.13C NMR (100 MHz, CD3OD): 6 181.6, 163.8, 104.7, 103.0, 81.4. 81.3, 77.7, 76.7, 75.1, 74.9, 74.7, 74.2, 71.5, 69.4, 62.8, 51.8, 33.1, 31.1, 31.0, 30.9, 30.8, 30.8, 30.1, 23.8, 14.6; HRMS (FAB.sup.+): For C.sub.45H.sub.80N.sub.4O.sub.22S.sub.2 [M+Na].sup.+ 1115.4603, found 1115.4601.

Preparation Example 29

Synthesis of TEM-T8

<29-1> Synthesis of Compound 3b

[0240] Compound 3b was synthesized in 84% yield according to Example 3-1. .sup.1H NMR (400 MHz, CDCl.sub.3): ? 4.42 (t, J=6.6 Hz, 4H), 1.81-1.77 (m, 4H), 1.44-1.30 (m, 20H), 0.89 (t, J=6.9 Hz, 6H); .sup.13C NMR (100 MHz, CDCl.sub.3): ? 172.7, 172.2, 69.6, 32.0, 29.7, 29.6, 28.6, 25.9, 22.8, 14.2.

<29-2> Synthesis of Compound 6b

[0241] Compound 6b was synthesized in 83% yield according to Example 3-2. .sup.1H NMR (400 MHz, CDCl.sub.3): ? 4.65 (bs, 2H), 3.92-3.90 (m, 4H), 3.81-3.79 (m, 4H), 3.03 (t, J=8.0 Hz, 4H), 1.71-1.67 (m, 4H), 1.42-1.27 (m, 20H), 0.88 (t, J=7.0 Hz, 6H); .sup.13C NMR (100 MHz, CDCl.sub.3): ? 179.8, 162.9, 62.0, 52.3, 32.0, 30.3, 29.6, 29.4, 29.1, 28.9, 22.6, 14.1.

<29-3> Synthesis of TEM-T8a

[0242] TEM-T8a was synthesized in 85% yield according to the general glycosylation reaction procedure of Example 3-3. .sup.1H NMR (400 MHz, CDCl.sub.3): ? 8.12 (d, J=7.2 Hz, 4H), 8.00 (d, J=7.2 Hz, 4H), 7.82 (d, J=7.2 Hz, 4H), 7.79-7.73 (m, 12H), 7.61 (d, J=7.3 Hz, 4H), 7.56-7.12 (m, 42H), 6.11 (t, J=10.0 Hz, 2H), 5.77-5.66 (m, 4H), 5.41-5.21 (m, 4H), 4.90 (d, J=10.5 Hz, 2H), 4.79-4.75 (m, 2H), 4.62-4.46 (m, 4H), 4.47-4.23 (m, 4H), 4.12-4.03 (m, 4H), 3.76 (t, J=7.2 Hz, 2H), 3.54-3.45 (m, 4H), 2.78 (t, J=7.2 Hz, 2H) 1.57-1.53 (m, 4H), 1.29-1.20 (m, 20H), 0.88 (t, J=7.0 Hz, 6H); .sup.13C NMR (100 MHz, CDCl.sub.3): ? 180.5, 179.8, 166.2, 165.9, 165.8, 165.5, 165.1, 164.8, 162.0, 134.0, 133.7, 133.4, 133.3, 130.1, 130.0, 129.9, 129.7, 129.5, 129.4, 129.2, 128.9, 128.7, 128.5, 128.3, 101.0, 95.6, 71.9, 71.3, 69.8, 69.1, 68.9, 63.2, 62.5, 32.0, 30.3, 29.7, 29.5, 29.3, 28.9, 22.7, 14.2.

<29-4> Synthesis of TEM-T8

[0243] TEM-T8 was synthesized in 94% yield according to the general synthesis procedure for the deprotection reaction of Example 3-4. .sup.1H NMR (400 MHz, CD.sub.3OD): ? 5.16 (d, J=7.0 Hz, 2H), 4.33 (d, J=8.0 Hz, 2H), 3.96-3.92 (m, 2H), 3.82-3.80 (m, 4H), 3.74-3.68 (m, 8H), 3.57-3.39 (m, 10H), 3.27-3.24 (m, 4H), 3.15-3.11 (m, 4H), 2.91 (t, J=7.2 Hz, 4H), 1.59-1.52 (m, 4H), 1.30-1.15 (m, 20H), 0.77 (t, J=7.0 Hz, 6H); .sup.13C NMR (100 MHz, CD.sub.3OD): ? 180.6, 163.0, 104.7, 103.0, 81.3, 77.9, 76.7, 75.2, 74.9, 74.8, 74.2, 71.5, 69.1, 62.8, 62.2, 33.1, 31.0, 30.5, 30.2, 23.8, 14.6; HRMS (FAB.sup.+): For C.sub.47H.sub.84N.sub.4O.sub.22S.sub.2 [M+Na].sup.+ 1143.4916, found 1143.4923. found 1129.4757.

Preparation Example 30

Synthesis of TEM-T9

<30-1> Synthesis of Compound 3c

[0244] Compound 3c was synthesized in 86% yield according to Example 3-1. .sup.1H NMR (400 MHz, CDCl.sub.3): ? 4.42 (t, J=6.6 Hz, 4H), 1.81-1.78 (m, 4H), 1.43-1.30 (m, 24H), 0.89 (t, J=6.9 Hz, 6H); .sup.13C NMR (100 MHz, CDCl.sub.3): ? 172.7, 172.2, 69.6, 32.0, 29.6, 29.4, 28.6, 25.9, 22.8, 14.2.

<30-2> Synthesis of Compound 6c

[0245] Compound 6c was synthesized in 80% yield according to Example 3-2. .sup.1H NMR (400 MHz, CDCl.sub.3): ? 4.65 (bs, 2H), 3.92-3.90 (m, 4H), 3.81-3.79 (m, 4H), 3.03 (t, J=8.0 Hz, 4H), 1.71-1.67 (m, 4H), 1.42-1.27 (m, 24H), 0.88 (t, J=7.0 Hz, 6H); .sup.13C NMR (100 MHz, CDCl.sub.3): ? 179.2, 162.1, 61.3, 52.8, 31.8, 30.1, 29.5, 29.2, 29.0, 28.9, 22.6, 14.1.

<30-3> Synthesis of TEM-T9a

[0246] TEM-T9a was synthesized in 82% yield according to the general glycosylation reaction procedure of Example 3-3. .sup.1H NMR (400 MHz, CDCl.sub.3): ? 8.12 (d, J=7.2 Hz, 4H), 8.00 (d, J=7.2 Hz, 4H), 7.8 (d, J=7.2 Hz, 4H), 7.79-7.73 (m, 12H), 7.6 (d, J=7.3 Hz, 4H), 7.56-7.12 (m, 42H), 6.11 (t, J=10.0 Hz, 2H), 5.77-5.66 (m, 4H), 5.41-5.21 (m, 4H), 4.90 (d, J=10.5 Hz, 2H), 4.79-4.75 (m, 2H), 4.62-4.46 (m, 4H), 4.47-4.23 (m, 4H), 4.12-4.03 (m, 4H), 3.76 (t, J=7.2 Hz, 2H), 3.54-3.45 (m, 4H), 2.78 (t, J=7.2 Hz, 2H) 1.57-1.53 (m, 4H), 1.29-1.20 (m, 24H), 0.88 (t, J=7.0 Hz, 6H); .sup.13C NMR (100 MHz, CDCl.sub.3): ? 180.5, 179.8, 166.2, 165.9, 165.8, 165.5, 165.1, 164.8, 162.0, 134.0, 133.7, 133.4, 133.3, 130.1, 130.0, 129.9, 129.7, 129.5, 129.4, 129.2, 128.9, 128.7, 128.5, 128.3, 101.0, 95.6, 71.9, 71.3, 69.8, 69.1, 68.9, 63.2, 62.5, 32.0, 30.3, 29.7, 29.5, 29.3, 28.9, 22.7, 14.2.

<30-4> Synthesis of TEM-T9

[0247] TEM-T9 was synthesized in 94% yield according to the general synthesis procedure for the deprotection reaction of Example 3-4. .sup.1H NMR (400 MHz, CD.sub.3OD): ? 5.16 (d, J=7.0 Hz, 2H), 4.34 (d, J=8.0 Hz, 2H), 3.95-3.91 (m, 2H), 3.82-3.79 (m, 4H), 3.75-3.65 (m, 8H), 3.54-3.38 (m, 10H), 3.32-3.22 (m, 4H), 3.15-3.10 (m, 4H), 2.91 (t, J=7.2 Hz, 4H), 1.59-1.52 (m, 4H), 1.30-1.15 (m, 24H), 0.76 (t, J=7.2 Hz, 6H); .sup.13C NMR (100 MHz, CD.sub.3OD): ? 180.4, 162.9, 104.7, 103.0, 81.3, 77.9, 76.7, 75.1, 74.8, 74.7, 74.2, 72.5, 69.1, 62.8, 62.2, 33.2, 31.1, 31.0, 30.8, 30.5, 30.1, 14.6; HRMS (FAB.sup.+): For C.sub.49H.sub.88N.sub.4O.sub.22S.sub.2 [M+Na].sup.+ 1171.5229, found 1171.5233.

Preparation Example 31

Synthesis of TEM-T10

<31-1> Synthesis of Compound 3d

[0248] Compound 3d was synthesized in 83% yield according to Example 3-1. .sup.1H NMR (400 MHz, CDCl.sub.3): ? 4.42 (t, J=6.6 Hz, 4H), 1.81-1.78 (m, 4H), 1.43-1.30 (m, 28H), 0.89 (t, J=6.9 Hz, 6H); .sup.13C NMR (100 MHz, CDCl.sub.3): ? 172.7, 172.2, 69.6, 64.3, 32.0, 29.7, 29.6, 29.4, 28.6, 25.9, 22.8, 14.2.

<31-2> Synthesis of Compound 6d

[0249] Compound 6d was synthesized in 82% yield according to Example 3-2. .sup.1H NMR (400 MHz, CDCl.sub.3): ? 3.92-3.90 (m, 4H), 3.80-3.78 (m, 4H), 3.02 (t, J=8.0 Hz, 4H), 1.70-1.67 (m, 4H), 1.42-1.27 (m, 28H), 0.88 (t, J=7.0 Hz, 6H); .sup.13C NMR (100 MHz, CDCl.sub.3): ? 179.2, 162.9, 52.4, 34.2, 32.1, 30.3, 29.7, 29.6, 29.5, 29.4, 29.2, 29.1, 22.8, 14.3.

<31-3> Synthesis of TEM-T10a

[0250] TEM-T10a was synthesized in 80% yield according to the general glycosylation reaction procedure of Example 3-3. .sup.1H NMR (400 MHz, CDCl.sub.3): ? 8.12 (d, J=7.2 Hz, 4H), 8.00 (d, J=7.2 Hz, 4H), 7.82 (d, J=7.2 Hz, 4H), 7.79-7.73 (m, 12H), 7.61 (d, J=7.3 Hz, 4H), 7.56-7.12 (m, 42H), 6.11 (t, J=10.0 Hz, 2H), 5.77-5.66 (m, 4H), 5.41-5.21 (m, 4H), 4.90 (d, J=10.5 Hz, 2H), 4.79-4.75 (m, 2H), 4.62-4.46 (m, 4H), 4.47-4.23 (m, 4H), 4.12-4.03 (m, 4H), 3.76 (t, J=7.2 Hz, 2H), 3.54-3.45 (m, 4H), 2.78 (t, J=7.2 Hz, 2H) 1.57-1.53 (m, 4H), 1.29-1.20 (m, 28H), 0.88 (t, J=7.0 Hz, 6H); .sup.13C NMR (100 MHz, CDCl.sub.3): ? 180.5, 179.8, 166.2, 166.1, 165.9, 165.5, 165.1, 164.9, 162.1, 134.1, 133.7, 133.5, 133.4, 133.2, 130.0, 129.8, 129.7, 129.5, 129.4, 129.3, 129.0, 128.9, 128.7, 128.5, 128.4, 101.0, 95.6, 75.5, 71.9, 71.3, 69.8, 69.1, 69.0, 63.2, 62.6, 32.0, 30.3, 30.1, 29.7, 29.4, 29.3, 28.9, 22.8, 14.2.

<31-4> Synthesis of TEM-T10

[0251] TEM-T10 was synthesized in 95% yield according to the general synthesis procedure for the deprotection reaction of Example 3-4. .sup.1H NMR (400 MHz, CD.sub.3OD): ? 5.17 (d, J=7.0 Hz, 2H), 4.35 (d, J=8.0 Hz, 2H), 3.95-3.91(m, 2H), 3.82-3.79 (m, 4H), 3.75-3.65 (m, 8H), 3.54-3.38 (m, 10H), 3.32-3.22 (m, 4H), 3.15-3.10 (m, 4H), 2.91 (t, J=7.2 Hz, 4H), 1.59-1.52 (m, 4H), 1.30-1.15 (m, 28H), 0.76 (t, J=7.0 Hz, 6H); .sup.13C NMR (100 MHz, CD.sub.3OD): ? 180.4, 163.2, 104.9 104.7, 103.0, 81.4, 81.3, 77.7, 76.7, 75.1, 74.9, 74.7, 74.2, 71.5, 62.8, 62.2, 33.2, 31.1, 31.0, 30.8, 30.6, 30.5, 30.1, 23.9 14.6; HRMS (FAB.sup.+): For C.sub.51H.sub.92N.sub.4O.sub.22S.sub.2 [M+Na].sup.+ 1199.5542, found 1199.5549.

Preparation Example 32

Synthesis of TEM-T11

<32-1> Synthesis of Compound 3e

[0252] Compound 3e was synthesized in 82% yield according to Example 3-1. .sup.1H NMR (400 MHz, CDCl.sub.3): ? 4.42 (t, J=6.8 Hz, 4H), 1.81-1.78 (m, 4H), 1.43-1.30 (m, 32H), 0.89 (t, J=6.9 Hz, 6H); .sup.13C NMR (100 MHz, CDCl.sub.3): ? 172.7, 172.2, 69.6, 32.0, 29.7, 29.6, 29.6, 29.3, 28.5, 25.8, 22.8, 14.2.

<32-2> Synthesis of Compound 6e

[0253] Compound 6e was synthesized in 84% yield according to Example 3-2. .sup.1H NMR (400 MHz, CDCl.sub.3): ? 4.62 (bs, 2H), 3.92-3.88 (m, 4H), 3.78-3.75 (m, 4H), 2.99 (t, J=7.4 Hz, 4H), 1.71-1.67 (m, 4H), 1.42-1.27 (m, 32H), 0.88 (t, J=7.0 Hz, 6H); .sup.13C NMR (100 MHz, CDCl.sub.3): ? 179.6, 162.6, 61.8, 52.6, 34.2, 32.1, 30.3, 29.8, 29.7, 29.6, 29.5, 29.2, 29.1, 28.5, 24.8, 22.6, 14.1.

<32-3> Synthesis of TEM-T11a

[0254] TEM-T11a was synthesized in 85% yield according to the general glycosylation reaction procedure of Example 3-3. .sup.1H NMR (400 MHz, CDCl.sub.3): ? 8.12 (d, J=7.2 Hz, 4H), 8.00 (d, J=7.2 Hz, 4H), 7.82 (d, J=7.2 Hz, 4H), 7.79-7.73 (m, 12H), 7.61 (d, J=7.3 Hz, 4H), 7.56-7.12 (m, 42H), 6.11 (t, J=10.0 Hz, 2H), 5.77-5.66 (m, 4H), 5.41-5.21 (m, 4H), 4.90 (d, J=10.5 Hz, 2H), 4.79-4.75 (m, 2H), 4.62-4.46 (m, 4H), 4.47-4.23 (m, 4H), 4.12-4.03 (m, 4H), 3.76 (t, J=7.2 Hz, 2H), 3.54-3.45 (m, 4H), 2.78 (t, J=7.2 Hz, 2H) 1.57-1.53 (m, 4H), 1.29-1.20 (m, 32H), 0.88 (t, J=7.0 Hz, 6H); .sup.13C NMR (100 MHz, CDCl.sub.3): ? 179.0, 171.1, 166.1, 165.8, 165.6, 165.4. 165.1, 165.0, 164.9, 161.2, 149.4, 133.4, 133.1, 130.0, 129.9, 129.8, 129.7, 129.6, 129.5, 129.3, 129.0, 128.9, 128.7, 128.6, 128.5, 128.4. 128.2. 128.1, 100.8, 96.3, 72.8, 72.7, 72.2, 69.9, 69.0, 68.3, 63.4, 62.5, 60.4, 53.5, 48.7, 31.9, 31.6, 30.0, 29.6, 29.4, 29.3, 29.0, 22.7, 21.0, 14.2.

<32-4> Synthesis of TEM-T11

[0255] TEM-T11 was synthesized in 92% yield according to the general synthesis procedure for the deprotection reaction of Example 3-4. .sup.1H NMR (400 MHz, CD.sub.3OD): 6 5.17 (d, J=7.0 Hz, 2H), 4.34 (d, J=8.0 Hz, 2H), 3.96-3.92 (m, 2H), 3.80-3.67 (m, 4H), 3.75-3.65 (m, 8H), 3.51-3.31 (m, 10H), 3.26-3.25 (m, 4H), 3.13-3.10 (m, 4H), 2.90 (t, J=7.2 Hz, 4H), 1.59-1.52 (m, 4H), 1.27-1.15 (m, 32H), 0.78 (t, J=7.0 Hz, 6H); .sup.13C NMR (100 MHz, CD.sub.3OD): ? 180.5, 163.0, 104.6, 103.0, 81.3, 77.9, 76.7, 75.1, 74.8, 74.7, 74.2, 72.5, 69.1, 62.8, 62.2, 33.2, 31.1, 31.0, 30.9, 30.8, 30.6, 30.5, 30.2, 23.9, 14.6; HRMS (FAB.sup.+): For C.sub.53H.sub.96N.sub.4O.sub.22S.sub.2 [M+Na].sup.+ 1227.5855, found 1227.5847.

Experimental Example 1

Properties of TSMs, TTGs and TEMs

[0256] To confirm the properties of TSMs, TTGs and TEMs synthesized by the synthetic methods of Examples 1 to 3, the molecular weights (MWs) and critical micelle concentrations (CMCs) of TSMs, TTGs and TEMs and the hydrodynamic radii (R.sub.h) of formed micelles were measured.

[0257] Specifically, the critical micelle concentrations (CMCs) were measured using a fluorescent dye, diphenylhexatriene (DPH), and the hydrodynamic radii (R.sub.h) of micelles formed by each preparation (1.0 wt %) were determined by a dynamic light scattering (DLS) experiment. The measured results are shown in Table 1 compared to DDM which is an existing amphiphilic molecule (detergent).

TABLE-US-00001 TABLE 1 detergent MW.sup.a CMC (mM) Rh.sup.b(nm) solubility TSM-E7 1047.1 ~0.04 3.4 ? 0.1 ~10% TSM-E8 1075.2 ~0.02 3.6 ? 0.1 ~10% TSM-E9 1103.2 ~0.01 4.0 ? 0.1 ~10% TSM-E10 1131.3 ~0.004 8.7 ? 0.7 ~5% TSM-E11 1159.3 ~0.002 56.2 ? 9.9 ~1% TSM-T7 1079.2 ~0.01 3.6 ? 0.1 ~10% TSM-T8 1107.3 ~0.006 4.2 ? 0.0 ~10% TSM-T9 1135.3 ~0.004 5.1 ? 0.1 ~10% TSM-T10 1163.4 ~0.003 38.2 ? 1.8 ~5% TSM-T11 1191.5 ~0.001 94.8 ? 45 ~1% TEM-E7 1093.3 ~0.03 3.3 ? .0 ~10% TEM -E8 1121.3 ~0.02 3.4 ? 0.1 ~10% TEM -E9 1149.4 ~0.009 3.9 ? 0.1 ~10% TEM -E10 1177.4 ~0.005 4.5 ? 0.1 ~5% TEM -E11 1205.5 ~0.001 25.2 ? 1.2 ~1% TEM -T7 1125.4 ~0.008 3.5 ? 0.0 ~10% TEM -T8 1153.4 ~0.004 3.6 ? 0.0 ~10% TEM -T9 1181.5 ~0.003 4.0 ? 0.1 ~10% TEM -T10 1209.5 ~0.0025 4.3 ? 0.0 ~5% TEM -T11 1237.6 ~0.001 31.8 ? 0.3 ~1% TTG-T7 947.11 ~0.02 2.8 ? 0.16 ~10% TTG-T8 975.17 ~0.015 2.9 ? 0.09 ~10% TTG-T9 1003.22 ~0.010 3.3 ? 0.11 ~10% TTG-T10 1031.28 ~0.006 3.6 ? 0.10 ~10% TTG-T11 1059.33 ~0.004 3.7 ? 0.09 ~10% TTG-T12 1087.38 ~0.0035 5.5 ? 0.20 ~5% TTG-A8 941.08 ~0.6 2.1 ? 0.56 ~10% TTG-A9 969.17 ~0.5 2.2 ? 0.20 ~10% TTG-A10 997.19 ~0.3 2.3 ? 0.14 ~10% TTG-A11 1025.25 ~0.2 2.4 ? 0.11 ~10% TTG-A12 1053.30 ~0.04 2.5 ? 0.07 ~5% TTG-A14 1109.41 ~0.008 3.2 ? 0.05 ~1% TTG-10, 10 1031.28 ~0.005 3.6 ? 0.01 ~10% TTG-9, 11 1031.28 ~0.004 3.7 ? 0.07 ~10% TTG-8, 12 1031.28 ~0.003 3.6 ? 0.07 ~10% TTG-6, 14 1031.28 ~0.002 3.6 ? 0.04 ~10% TTG-4, 16 1031.28 ~0.001 3.6 ? 0.03 ~10% TTG-11, 11 1059.34 ~0.004 3.7 ? 0.09 ~10% TTG-10, 12 1059.34 ~0.003 3.6 ? 0.06 ~10% TTG-8, 14 1059.34 ~0.003 2.6 ? 0.02 ~10% TTG-6, 16 1059.34 ~0.002 4.0 ? 0.04 ~10% DDM 510.62 ~0.17 3.4 ? 0.03 ~10%

[0258] The CMC values (0.001 to 0.6 mM) of most TSMs, TTGs and TEMs except for some TTG-As compounds were significantly lower than the CMC value (0.17 mmM) of DDM. Therefore, TSMs, TTGs and TEMs easily form micelles even at low concentrations, indicating a higher tendency to self-assemble. In addition, the CMC values of TSMs, TTGs and TEMs decreased with increasing alkyl chain length, which is determined to be due to the increase in hydrophobicity as the alkyl chain length increases. Furthermore, compounds including a thioether (TSM-Ts TTG-Ts and TEM-Ts) have lower CMC values than compounds including an ether (TSM-Es and TEM-Es), which is determined to be because a thioether exhibits higher hydrophobicity than an ether. The sizes of micelles formed by TSMs, TTGs and TEMs generally showed a tendency to increase as the alkyl chain length increased. This is due to a change in geometry of amphiphilic molecules from a conical to cylindrical shape as the alkyl chain length increases. It was confirmed that TTGs form micelles that are generally smaller than TEMs and TSMs. Meanwhile, as a result of examining the size distribution of micelles formed by TSMs and TEMs through DLS, the number- and volume-weighted size distributions both showed a single set of populations, indicating high homogeneity (FIGS. 4 and 5).

[0259] From these results, it could be confirmed that the TSMs and TEMs of the present invention have lower CMC values than DDM, and thus have a much higher tendency to self-assemble because micelles are easily formed even in small amounts, that the sizes of micelles formed by TSMs and TEMs differ depending on the structure of a linker and the type of atom (oxygen or sulfur) constituting the linker, and that the micelles formed by TSMs and TEMs have high homogeneity.

Experimental Example 2

Evaluation of Ability of TSMs, TTGs and TEMs to Stabilize Structure of LeuT Membrane Protein

[0260] Experiments were performed to measure the structural stability of the LeuT protein by TSMs, TTGs and TEMs. Each amphiphilic compound was used at a concentration of (a) CMC+0.04 wt % or (b) CMC+0.2 wt %, and the substrate binding properties of LeuT were measured using [.sup.3H]-Leu via scintillation proximity assay (SPA). The measurement was performed at regular intervals over the course of a 13-day and 12-day incubation at room temperature, respectively.

[0261] Specifically, a wild-type leucine transporter (LeuT) derived from the thermophilic bacterium Aquifex aeolicus was purified by a previously described method (Nature 1998, 392, 353-358 by G. Deckert et al.). LeuT was expressed in E. coli C41 (DE3) transformed with pET16b encoding the C-terminal 8?His-tagged transporter (expression plasmids were provided from Dr E. Gouaux, Vollum Institute, Portland, Oregon, USA). In summary, after a bacterial membrane was isolated and solubilized in 1 wt % DDM, the protein was bound to Ni.sup.2+-NTA resin (Life Technologies, Denmark), and eluted in 20 mM Tris-HCl (pH 8.0), 1 mM NaCl, 199 mM KCl, 0.05%(w/v) DDM and 300 mM imidazole. Then, the purified LeuT (0.5 mg/ml) was diluted with buffers supplemented with TSMs, TTGs, TEMs or DDM at a final concentration of CMC+0.04% (w/v) or CMC+0.2% (w/v) except for DDM and imidazole in the equivalent buffer described above. Protein samples were incubated at room temperature for 13 days, centrifuged at a specified time, and protein properties were confirmed by measuring [.sup.3H]-Leucine binding ability using SPA. SPA was performed with 5 ?L of each protein sample in a buffer containing 450 mM NaCl and each of TSMs and TEMs (or DDM). The SPA reaction was performed in the presence of 20 nM [.sup.3H]-Leucine and 1.25 mg/ml copper chelate (His-Tag) YSi beads (PerkinElmer, Denmark). Overall [.sup.3H]-Leucine coupling to each sample was measured using a MicroBeta liquid scintillation counter (Perkin Elmer).

[0262] As illustrated in FIGS. 6 to 8, all TSMs, TTGs and TEMs had an excellent effect of maintaining the substrate binding properties of LeuT during a 13-day incubation period. That is, most TSMs, TTGs and TEMs completely retained the transporter substrate binding properties for a long time even at high compound concentrations. In contrast, it was confirmed that DDM had high initial activity, but the substrate binding ability sharply decreased with the passage of time. In particular, the best substrate binding activity was confirmed in compounds having intermediate alkyl chain lengths (TSM-E8/T9/T10, TTG-T10/T11/T12 and TEM-E10/T8/T9/T10), whereas compounds having short alkyl chain lengths are excellent in terms of long-term stability. Further, it was confirmed that TEMs generally have an excellent effect on maintaining LeuT stability compared to TSMs and TTGs.

[0263] It was confirmed that, at CMC+0.2 wt %, most TSMs, TTGs and TEMs had low initial activity, but were effective in maintaining the initial activity for a long period of time, compared to DDM, and after a certain period of time passed, most TSMs, TTGs and TEMs exhibited excellent activity compared to DDM.

[0264] These results suggest that the types of structural linkers (serinol or diethanolamine) and the alkyl chain lengths of overall TSMs and TEMs acted as important factors in maintaining LeuT structural stability.

Experimental Example 3

Evaluation of Ability of TSMs, TTGs and TEMs to Stabilize Structure of MelB Membrane Protein

[0265] Experiments were performed to measure the structural stability of Salmonella typhimurium melibiose permease (MelBst) protein by TSMs, TTGs and TEMs. After the MelB protein was extracted from the membrane using TSMs, TTGs, TEMs or DDM, the amount and structure of the extracted protein were analyzed by SDS-PAGE and Western blotting. The concentration of the amphiphilic compound used was 1.5 wt %, the protein was extracted at 0? C. for 90 minutes and then the extracted protein was incubated at three high temperatures (45, 55, and 65? C.) for an additional 90 minutes. By measuring the amount of protein remaining in a dissolved state in an aqueous solution of the extracted protein and the heat-treated protein, we tried to evaluate the performance of both the protein extraction efficiency and stabilization ability of the compound at the same time. The amount of protein extracted and stabilized by each amphiphilic molecule is shown as a value (%) relative to the amount of total protein contained in the membrane sample untreated with the amphiphilic molecule.

[0266] Specifically, Salmonella typhimurium melibiose permease (MelB.sub.St) having a 10-His tag at the C-terminus was expressed in E. coli DW2 cells (?melB and ?lacZY) using a plasmid pK95?AHB/WT MelB.sub.St/CH10. Cell growth and membrane preparation were performed according to the method described in the paper by A. S. Ethayathulla et al. (Nat. Commun. 2014, 5, 3009). Protein assay was performed with a Micro BCA kit (Thermo Scientific, Rockford, IL). TSMs, TEMs or DDM were evaluated for MelBst stability using the protocol described in Nat. Methods 2010, 7, 1003-1008 by P. S. Chae et al. A membrane sample containing MelB.sub.St (final protein concentration was 10 mg/mL) was incubated in a solubilizing buffer (20 mM sodium phosphate, pH 7.5, 200 mM NaCl, 10% glycerol, and 20 mM melibiose) containing 1.5% (w/v) DDM, TSMs, TTGs or TEMs at four temperatures (0, 45, 55, and 65? C.) for 90 minutes. To remove insoluble materials, ultracentrifugation was performed at 355,590 g using a Beckman Optima? MAX ultracentrifuge equipped with a TLA-100 rotor at 4? C. for 45 minutes. The solubilized portion was isolated by SDS-16% PAGE, and then immunoblotted with a HisProbe-HRP antibody (Thermo Scientific). A membrane fraction containing 20 ?g of the protein without any treatment was used to show the entire MelB, and treated samples were loaded into each well in equivalent volumes. MelB.sub.St was measured by an ImageQuant LAS 4000 Biomolecular Imager (GE Health Care Lifer Science) using a SuperSignal West Pico chemiluminescent substrate.

[0267] As shown in the results illustrated in FIGS. 9 to 11, DDM showed high MelB protein extraction efficiency at 0? C. compared to all TEMs, TTGs and TSMs except for TEM-E8.

[0268] However, it could be confirmed that when the temperature was raised to 45? C., the ability of DDM to solubilize the MelB protein deteriorated, whereas all TEMs had enhanced MelB protein-solubilizing ability compared to 0? C., and TEMs (E7/E8/E9/T7/T8/T9) showed the ability to maintain a better protein solubilization state than DDM. In the case of TSMs, like TEMs, MelB protein solubilization efficiency increased at 45? C. compared to 0? C., but was lower than that of DDM. It could be confirmed that the ability of TTGs to solubilize the MelB protein was enhanced compared to 0? C., and TTGs (T9/T10) exhibited a better ability to solubilize the protein than DDM.

[0269] When the temperature was raised to 55? C., the ability of DDM to solubilize the protein remarkably deteriorated, but most TEMs, TTGs and TSMs showed a better ability to maintain the MelB protein solubilization state than DDM. At a temperature of 65? C., no solubilized MelB protein was identified in all of TEMs, TTGs, TSMs and DDM.

[0270] As a whole, it could be confirmed that at a low temperature (0? C.), DDM showed higher protein extraction efficiency than TEMs, TTGs and TSMs, whereas as the temperature increased (45? C. and 55? C.), the amount of MelB protein solubilized by TEMs, TTGs and TSMs was increased compared to that of DDM, and therefore, it was confirmed that DDM has excellent protein extraction efficiency, but TEMs, TTGs and TSMs are better in the ability to maintain the protein solubilization state, that is, the ability to stabilize the protein.

Experimental Example 4

Evaluation of Ability of TSMs, TTGs and TEMs to Structurally Stabilize ?.SUB.2.AR Membrane Proteins

[0271] Experiments were performed to measure the structural stability of the human ?.sub.2 adrenergic receptor (?.sub.2AR) and G-protein-coupled receptor (GPCR) by TSMs, TTGs and TEMs. That is, a receptor purified by DDM was subjected to amphiphilic molecular exchange when diluted with a buffer solution containing only each of TSMs, TTGs and TEMs without cholesteryl hemisuccinate (CHS) or a buffer solution containing CHS and DDM. The final concentration of the amphiphilic molecule was CMC+0.2wt %, and the ligand binding properties of the receptor were measured by binding of [.sup.3H]-dihydroalprenolol ([.sup.3H]-DHA).

[0272] Specifically, the following method was used for a radioligand binding test. ?.sub.2AR was purified using 0.1% DDM (D. M. Rosenbaum et al., Science, 2007, 318, 1266-1273.) and finally concentrated to about 10 mg/ml (about 200 ?M). A master binding mixture containing 10 nM [.sup.3H]-dihydroalprenolol (DHA) supplemented with 0.5 mg/ml BSA in 0.2% amphiphilic compounds (DDM, TSMs, TTGs or TEMs) was prepared using ?.sub.2AR purified with DDM. Receptors purified with DDM, TSMs, TTGs or TEMs were incubated with 10 nM [.sup.3H]-DHA at room temperature for 6 days. The mixture was loaded onto a G-50 column and a flow through was collected with 1 ml of a binding buffer (20 mM HEPES supplemented with 0.5 mg/ml BSA and 20?CMC each amphiphilic compound, pH 7.5, 100 mM NaCl), and filled with 15 ml of a scintillation fluid. Receptor-bound [.sup.3H]-DHA was measured by a scintillation counter (Beckman). The binding of [.sup.3H]-DHA is shown as a column graph.

[0273] As illustrated in FIGS. 12 to 14, in the ligand binding properties of the receptor immediately after the amphiphilic molecular exchange, most TSMs, TTGs and TEMs were similar or superior compared to DDM (FIGS. 12 and 14A).

[0274] Furthermore, a test to confirm long-term ligand binding retention properties of the receptor were performed on TSMs (E10/E11/T7/T8/T9), TTGs (T9/T10/T11/T12) and TEMs (T8/T9), which have excellent effects of the above results. Specifically, ligand binding properties for receptors dissolved in TSMs (E10/E11/T7/T8/T9), TTGs (T9/T10/T11/T12), TEMs (T8/T9) or DDM were monitored at regular intervals while being incubated at room temperature for 6 days, and the results are illustrated in FIGS. 13 and 14B. As a result, TSMs (E10/E11/T7/T8/T9), TTGs (T9/T10/T11/T12) and TEMs (T8/T9) were better in maintaining the ligand binding ability of the receptor than DDM.

Experimental Example 5

Ability of TSMs and TEMs to Stabilize AtBOR1 Protein

[0275] Fluorescence size exclusion chromatography (FSEC): BOR1 of Arabidopsis thaliana was expressed in Saccharomyces cerevisiae FGY217 cells as a fusion protein having a C-terminal GFP tag. The cells were grown in a URA medium supplemented with 0.1% glucose. Protein expression was induced by adding 2% galactose, and then the cells were cultured at 20? C. for 18 hours as previously described. The cells were collected and used to prepare a membrane. The membrane including the BOR1-GFP fusion protein was diluted as follows. The membrane was diluted with PBS (pH 7.4) supplemented with 1% DDM or 1% individual TSM (TSM-E9/E10/T8/T9) and TEM (TEM-E9/E10/T8/T9) so that a final total protein concentration was 2.8 mg/ml. Samples were cultured while shaking at 4? C. for 1 hour, and then insoluble materials were removed by centrifugation at 14,000 g and 4? C. for 1 hour. A supernatant including the solubilized protein sample was heated at 47? C. for 10 minutes. After additional centrifugation to remove large aggregates, a 200 ?l aliquot of the sample was injected into a Superose 6 10/300 column equilibrated with 20 mm Tris (pH 7.5), 150 mm NaCl and 0.03% DDM. The GFP fluorescence of each fraction was read using an excitation wavelength of 470 nm and an emission wavelength of 512 nm.

[0276] CPM assay: AtBOR1 was concentrated to 10 mg/ml in a buffer (20 mM Tris-HCl (pH 7.5), 150 mM NaCl, 0.03% DDM) with a 100 kDa MWCO centrifugal filter, stored at ?80? C. and thawed immediately prior to thermal stability analysis. An analytical solution was prepared in a 96-well plate so that a final volume became 150 ?L with each detergent (TSM-E9/E10/T8/T9 and TEM-E9/E10/T8/T9) at CMC+0.04 or CMC+0.2 wt %, 20 mM Tris-HCl (pH 7.5), 150 mM NaCl and 1 ?L of AtBOR1. A 7-diethylamino-3-(4-maleimidylphenyl)-4-methylcoumarin (CPM) dye (Invitrogen) was dissolved in DMSO at 4 mg/ml, and was diluted 1:100 with a buffer (150 mM NaCl supplemented with 20 mM Tris-HCl (pH 7.5) and 0.03% DDM). After 3 ?L of the CPM dye was added to each well in the dark, the plate was covered with a transparent lid and the cells were cultured at 40? C. for 120 minutes. Fluorescence emission was monitored every 5 minutes using SpectraMax M2 (Molecular Devices) with a transition wavelength of 387 nm and an emission wavelength of 463 nm. Fluorescence readings were normalized under the most unstable conditions to calculate the percentage of proteins that were relatively unfolded. Data was analyzed as a single exponential decay curve using GraphPad Prism 6.

[0277] When the selected amphiphilic molecules were tested at CMC+0.04 wt %, all compounds were much better than DDM in their ability to preserve AtBOR1 in the folded state, and the best performance was observed, particularly, in TEM-E10 (FIGS. 15A and 15B). Further, similar results were confirmed even at an increased amphiphilic molecular concentration of CMCs+0.2 wt % (FIG. 16). As a result of further evaluating DDM or TEMs (TEM-E9/E10/T8/T9) using fluorescence size exclusion chromatography (FSEC), the protein solubilized with DDM showed a significant reduction in the original homogeneous protein peak (fraction number 35), along with a large increase in the peak caused by protein aggregation (fraction number 2) (FIG. 15C). In contrast, all tested TEMs were highly effective in maintaining the initial state of the protein, because the peaks caused by aggregation significantly reduced and the amount of decrease in the original protein peak was significantly decreased under the same conditions (FIGS. 15C and 15D). These results indicate that the compound of the present invention is less efficient in AtBOR1 extraction than DDM, but is better than DDM in thermal stabilization.

Experimental Example 6

MOR Thermal Stability Test

[0278] An N-[4-(7-diethylamino-4-methyl-3-coumarinyl)phenyl]maleimide (CPM) dye dissolved in DMSO (3 mg/ml) was diluted 40? in a buffer including 20 mM HEPES pH 7.5 and 150 mM NaCl. A ?-opioid receptor)(about 4 ?M) dissolved in DDM (0.05%)/CHS (0.005%) was cultured with 250 L of a 1.0% compound (TSM-E9/E10/T8/T9 and TEM-E9/E10/T8/T9) solution. After 1 hour at room temperature, a compound was prepared at a final concentration of 0.5 wt % by diluting the receptor solution 2? in 20 mM HEPES pH 7.5, 150 mM NaCl. After 5 L of the diluted CPM dye was added thereto, the receptor stability of different compounds was measured by recording the fluorescence spectra (excitation 387 nm) every 5? C. from 20? C. to 65? C. while culturing at each temperature for 2 minutes. The melting point (Tm) was calculated by plotting the read value at 470 nm and fitting a non-linear regression curve using GraphPad Prism.

[0279] As a result, the receptor dissolved in DDM/CHS showed a low melting point of 31.6? C. (FIG. 17). In contrast, all tested TSM/TEMs resulted in higher receptor melting points than DDM. TEM-T8, the least effective of the tested compounds, also showed a melting point of 35.9? C., which is 4.3? C. higher than that of DDM. TEM-E10 and TSM-E10 showed the highest melting points of 48.0? C. and 48.9? C., respectively, and these values are higher than that (38.3? C.) of LIVING, which is an optimized novel compound contributing significantly to the structure determination of many GPCRs.