KR20190119543A - Novel steroid -based amphiphiles and uses thereof - Google Patents

Abstract

The present invention relates to a newly developed steroid-based amphiphilic compound, a preparation method thereof and a method for extracting, solubilizing, stabilizing, crystallizing or analyzing membrane proteins using the same. In addition, the compound, compared to the conventional compounds, can efficiently extract membrane proteins having various structures and properties from cell membranes and store the same in an aqueous solution for a long time, and thus can be used for functional analysis and structural analysis. Membrane protein structure and function analysis thereof is one of the areas of greatest interest in current biology and chemistry by being closely related to drug development.

Description

Novel steroid-based amphiphiles and uses approximately

The present invention relates to a newly developed steroid-based amphiphilic compound, a preparation method thereof and a method for extracting, solubilizing, stabilizing, crystallizing or analyzing membrane proteins using the same.

Membrane proteins play an important role in biological systems. Since these bio-macromolecules contain hydrophilic and hydrophobic moieties, amphiphilic molecules are required to extract membrane proteins from cell membranes and to solubilize and stabilize them in aqueous solutions.

For the structural analysis of membrane proteins, it is necessary to obtain high-quality membrane protein crystals, which requires the structural stability of membrane proteins in aqueous solution. There are more than 100 existing amphiphilic molecules that have been used for membrane protein research, but only about 5 of them have been actively used for membrane protein structure research. These five amphoteric molecules are n-octyl-β-D-glucopyranoside (OG), n-nonyl-β-D-glucopyranoside (NG), n-decyl-β-D-maltopyranoside (DM), and DDM (n- dodecyl-β-D-maltopyranoside) and LDAO (lauryldimethylamine- N- oxide) are included (nonpatent literature 1, nonpatent literature 2). However, many membrane proteins surrounded by these molecules have significant limitations in studying the function and structure of membrane proteins utilizing these molecules because their structure is easily denatured or aggregated and tends to lose their function quickly. This is because conventional molecules do not exhibit various characteristics due to their simple chemical structure. Thus, there is a need for the development of new amphoteric materials with new and superior properties through new structures.

The present inventors have developed an amphiphilic compound having a steroid hydrophobic group, and completed the present invention by confirming the membrane protein stabilization properties of the compound.

 S. Newstead et al., Protein Sci. 17 (2008) 466-472.  S. Newstead et al., Mol. Membr. Biol. 25 (2008) 631-638.

An object of the present invention is to provide a compound represented by any one of formulas (1) to (3).

Another object of the present invention is to provide a composition for extraction, solubilization, stabilization, crystallization or analysis of a membrane protein comprising the compound.

Another object of the present invention is to provide a method for preparing the compound.

Still another object of the present invention is to provide a method for extracting, solubilizing, stabilizing, crystallizing or analyzing membrane proteins using the compounds.

One embodiment of the present invention provides a compound represented by any one of the following Formula 1 to Formula 3:

[Formula 1]

[Formula 2]

[Formula 3]

In Chemical Formulas 1 to 3,

Dotted lines in the formulas may represent a single bond or a double bond;

또는

로, 상기 n은 1, 2, 3, 4 또는 5인 정수일 수 있고, 상기 X⁴는 산소와 연결된 당류(saccharide)일 수 있으며;L ¹ is a direct bond,

or

N may be an integer of 1, 2, 3, 4 or 5, and X ⁴ may be a saccharide linked to oxygen;

X ² and X ³ may each independently be a saccharide connected with hydrogen or oxygen, and

X ¹ may be a saccharide connected with oxygen.

As used herein, the term "saccharide" refers to a compound that is relatively small molecule in carbohydrates and is sweet in water. Sugars are classified into monosaccharides, disaccharides and polysaccharides according to the number of molecules constituting the sugar.

The sugars used in the above embodiments may be monosaccharides, disaccharides, or pentasaccharides, specifically glucose, maltose or glucose centered pentasaccharides, but are not limited thereto. It doesn't work.

First, the steroid-based hydrophobic group of the present invention is characterized by a large number of rings connected in a plate shape and a high surface area and a high hydrophobic density, which can directly affect membrane protein stabilization by contributing to the hydrophobic effect. Amphiphilic molecules with a good hydrophobic-hydrophilic balance were designed by attaching two maltose with high hydrophilic density or using high density saccharides.

The compound according to an embodiment of the present invention may have a linker connecting a steroidal compound which is a hydrophobic group and a saccharide which is a hydrophilic group. The present inventors designed to flexibly interact with membrane proteins by controlling the length of the linker to control the overall flexibility of the amphipathic molecules. The linker may be a substituted or unsubstituted alkylene group, the substituted alkylene group may be a saccharide is bonded.

Specifically, any one of the compounds of Formulas 1 to 3 wherein L ¹ is a direct bond; One of X ² and X ³ is hydrogen and the other is a saccharide linked with oxygen; And X ¹ may be a saccharide connected with oxygen.

이고, 상기 n은 1, 2, 3, 4 또는 5인 정수이며; 상기 X² 및 X³은 수소이며; 그리고 상기 X¹은 산소와 연결된 당류일 수 있다.Specifically, the compound of any one of Formulas 1 to 3 is L ¹

N is an integer of 1, 2, 3, 4 or 5; X ² and X ³ are hydrogen; And X ¹ may be a saccharide connected with oxygen.

이고, 상기 n은 1, 2, 3, 4 또는 5인 정수이고, 상기 X⁴는 산소와 연결된 당류(saccharide)이며; 상기 X² 및 X³은 수소이고; 그리고 상기 X¹은 산소와 연결된 당류일 수 있다.Specifically, the compound of any one of Formulas 1 to 3 is L ¹

N is an integer equal to 1, 2, 3, 4 or 5, and X ⁴ is a saccharide linked to oxygen; X ² and X ³ are hydrogen; And X ¹ may be a saccharide connected with oxygen.

In one embodiment of the present invention, in Chemical Formulas 1 to 3, L ¹ is a direct bond; One of X ² and X ³ is hydrogen and the other is a saccharide linked with oxygen; And X ¹ is a compound which may be glucose linked with oxygen was named "GDNs (glyco-diosgenin analogs)".

이고, 상기 n은 1, 2, 3, 4 또는 5인 정수이며; 상기 X² 및 X³ 은 수소이며; 그리고 상기 X¹은 산소와 연결된 글루코스 중심의 가지친 오당류인 화합물을 각각 "SPSs (steroid-based penta-saccharides)" 또는 "SPS-Ls (steroid-based penta-saccharides with linker)"로 명명하였다.In another embodiment of the present invention, in Chemical Formulas 1 to 3, L ¹ is a direct bond or

N is an integer of 1, 2, 3, 4 or 5; X ² and X ³ are hydrogen; In addition, the X ¹ is a compound that is a branched saccharide of glucose centered with oxygen, respectively, named "SPSs (steroid-based penta-saccharides)" or "SPS-Ls (steroid-based penta-saccharides with linker").

이고, 상기 n은 1, 2, 3, 4 또는 5인 정수이고, 상기 X⁴는 산소와 연결된 말토오스이며; 상기 X² 및 X³은 수소이고; 그리고 상기 X¹은 산소와 연결된 말토오스인 화합물을 "SMAs (Steroids-based maltosides amphiphiles)"로 명명하였다.In another embodiment of the present invention, in Chemical Formulas 1 to 3, L ¹ is L ¹ is

N is an integer equal to 1, 2, 3, 4 or 5, and X ⁴ is maltose linked with oxygen; X ² and X ³ are hydrogen; In addition, X ¹ is a compound which is maltose linked with oxygen, named as "SMA (Steroids-based maltosides amphiphiles)".

In one embodiment of the present invention, the compound may be one selected from Formula 4 to Formula 22:

[Formula 4]

[Formula 5]

[Formula 6]

[Formula 7]

[Formula 8]

[Formula 9]

[Formula 10]

[Formula 11]

[Formula 12]

[Formula 13]

[Formula 14]

[Formula 15]

[Formula 16]

[Formula 17]

[Formula 18]

 [Formula 19]

[Formula 20]

[Formula 21]

[Formula 22]

Compounds according to other embodiments of the invention may be, but are not limited to, amphiphilic molecules for extracting, solubilizing, stabilizing, crystallizing or analyzing membrane proteins.

Specifically, the extraction may be to extract the membrane protein from the cell membrane.

As used herein, the term "amphiphilic molecule" refers to a molecule in which a hydrophobic group and a hydrophilic group coexist in one molecule to have affinity for both polar and nonpolar solvents. Phospholipid molecules present in surfactants and cell membranes are amphiphilic molecules with hydrophilic groups at one end and hydrophobic groups at the other end, and have the characteristic of forming micelles or liposomes in aqueous solution. Since hydrophilic groups have polarity but nonpolar groups coexist, their amphiphilic molecules tend to be insoluble in aqueous solutions. However, when the concentration is above a certain limit concentration (critical micelle concentration, CMC), hydrophobic interaction causes hydrophobic groups to collect inside and round or elliptic micelles with hydrophilic groups exposed to the surface, which greatly increases the solubility in water. .

The method of measuring CMC is not particularly limited, but a method well known in the art may be used, and for example, may be measured by a fluorescence staining method using diphenylhexatriene (DPH).

The compound according to an embodiment of the present invention may have a critical micelle concentration (CMC) in an aqueous solution of 0.0001 to 1 mM, specifically 0.0001 to 0.5 mM, more specifically 0.0005 to 0.4 mM, but is not limited thereto. I never do that.

In the case of DDM, which is mainly used for membrane protein research, most of the SPSs and SPS-Ls of this embodiment have a small CMC value compared to the critical micelle concentration of 0.17 mM. Therefore, since micelles are easily formed even at low concentrations, SPSs or SPS-Ls can be effectively studied and analyzed using a small amount, which is advantageous in terms of utilization than DDM.

In addition, another embodiment of the present invention provides a composition for extraction, solubilization, stabilization, crystallization or analysis of membrane proteins comprising the compound.

Specifically, the extraction may be to extract the membrane protein from the cell membrane.

As used herein, the term "extraction of membrane proteins" refers to the separation of membrane proteins from membranes.

As used herein, the term "solubilization" of membrane proteins means that membrane proteins that are insoluble in water are dissolved in micelles in aqueous solution.

As used herein, the term "stabilization of membrane proteins" means the stable preservation of tertiary or quaternary structures such that the structure, function of the membrane protein does not change.

As used herein, the term "crystallization of membrane proteins" means the formation of crystals of membrane proteins in solution.

As used herein, the term "analysis" of a membrane protein means to analyze the structure or function of the membrane protein. In the above embodiment, the analysis of the membrane protein may use a known method, but is not limited thereto. For example, the structure of the membrane protein may be analyzed using electron microscopy or nuclear magnetic resonance. can do.

The composition may be, but is not limited to, a formulation of micelles, liposomes, emulsions or nanoparticles.

The micelle may have a radius of 2.0 nm to 10 nm, and specifically 2.0 nm to 6.0 nm, but is not limited thereto.

The method of measuring the radius of the micelle is not particularly limited, but a method well known in the art may be used, and may be measured using, for example, dynamic light scattering (DLS) experiments.

The micelles, liposomes, emulsions or nanoparticles can be combined with the membrane protein hydrophobic inside. That is, the micelles, liposomes, emulsions or nanoparticles can be wrapped by extracting the membrane protein present in the cell membrane. Therefore, it is possible to extract, solubilize, stabilize, crystallize or analyze membrane proteins from cell membranes by the micelles.

The composition may further include a buffer or the like that may be helpful for extraction, solubilization, stabilization, crystallization or analysis of the membrane protein.

Further, another embodiment of the present invention provides a method for preparing a compound represented by one of the following Chemical Formulas 1 to 3, including the steps of 1) to 3):

1) Cis or trans hydrides on carbons 2 and 3 of the steroids of ticogenin, diosgenin, cholesterol, Cholesterol, Cholestanol or Sitosterol Introducing oxy;

2) introducing a saccharide with a protecting group by performing a glycosylation reaction on the product of step 1); And

3) performing a deprotection reaction on the product of step 2);

[Formula 1]

[Formula 2]

[Formula 3]

In Chemical Formulas 1 to 3,

Dotted line in the formula represents a single bond or a double bond,

L ¹ is a direct bond;

X ¹ and X ² are sugars connected with oxygen, and X ³ is hydrogen.

The compound synthesized by the method may be one compound of Formulas 4 to 5 according to an embodiment of the present invention, but is not limited thereto.

In this embodiment, since the compound can be synthesized by a simple method through three steps of synthesis, the compound for membrane protein research can be easily and economically produced.

Further, another embodiment of the present invention provides a method for preparing a compound represented by one of the following Chemical Formulas 1 to 3, including the steps of 1) to 3):

1) introducing hydroxy to carbon 3 and 6 of the steroid of ticogenin, diosgenin, cholesterol (Cholesterol), cholestanol (Cholestanol) or cytosterol (Sitosterol);

2) introducing a saccharide with a protecting group by performing a glycosylation reaction on the product of step 1); And

3) performing a deprotection reaction on the product of step 2);

[Formula 1]

[Formula 2]

[Formula 3]

In Chemical Formulas 1 to 3,

Dotted line in the formula represents a single bond or a double bond,

L ¹ is a direct bond;

X ¹ and X ³ are saccharides connected with oxygen, and X ² is hydrogen.

The compound synthesized by the above method may be a compound represented by Chemical Formula 6 according to an embodiment of the present invention, but is not limited thereto.

Further, another embodiment of the present invention provides a method for preparing a compound represented by one of the following Chemical Formulas 1 to 3, including the steps of 1) to 3):

1) Introducing a substituted or unsubstituted alkylene linker to the hydroxy of carbon 3 of the steroid of ticogenin, diosgenin, cholesterol, Cholesterol or Citostanol Or performing a glycosylation reaction to introduce a saccharide to which a protecting group is attached;

2) introducing a saccharide to which a protecting group is attached by performing a glycosylation reaction on the product into which the alkylene group linker of step 1) is introduced; And

3) performing a deprotection reaction on the glycosylation product of step 1) or the product of step 2);

[Formula 1]

[Formula 2]

[Formula 3]

In Chemical Formulas 1 to 3,

또는

로, 상기 n은 1, 2, 3, 4 또는 5인 정수이고, 상기 X⁴는 산소와 연결된 당류(saccharide)이며;L ¹ is a direct bond,

or

N is an integer of 1, 2, 3, 4 or 5, and X ⁴ is a saccharide linked to oxygen;

X ² and X ³ are hydrogen; And

X ¹ is a saccharide connected with oxygen.

The compound synthesized by the above method may be a compound of Formula 7 to 22 according to one embodiment of the present invention, but is not limited thereto.

Another embodiment of the present invention provides a method for extracting, solubilizing, stabilizing, crystallizing or analyzing membrane proteins. Specifically, the present invention provides a method for extracting, solubilizing, stabilizing, crystallizing, or analyzing a membrane protein, comprising treating the membrane protein with a compound represented by one of the following Chemical Formulas 1 to 3 in an aqueous solution:

[Formula 1]

[Formula 2]

[Formula 3]

In Chemical Formulas 1 to 3,

Dotted lines in the formulas may represent a single bond or a double bond;

또는

로, 상기 n은 1, 2, 3, 4 또는 5인 정수일 수 있고, 상기 X⁴는 산소와 연결된 당류(saccharide)일 수 있으며;L ¹ is a direct bond,

or

N may be an integer of 1, 2, 3, 4 or 5, and X ⁴ may be a saccharide linked to oxygen;

X ² and X ³ may each independently be a saccharide connected with hydrogen or oxygen, and

X ¹ may be a saccharide connected with oxygen.

Specifically, any one of the compounds of Formulas 1 to 3 wherein L ¹ is a direct bond; One of X ² and X ³ is hydrogen and the other is a saccharide linked with oxygen; And X ¹ may be a saccharide connected with oxygen.

이고, 상기 n은 1, 2, 3, 4 또는 5인 정수이며; 상기 X² 및 X³은 수소이며; 그리고 상기 X¹은 산소와 연결된 당류일 수 있다.Specifically, the compound of any one of Formulas 1 to 3 is L ¹

N is an integer of 1, 2, 3, 4 or 5; X ² and X ³ are hydrogen; And X ¹ may be a saccharide connected with oxygen.

이고, 상기 n은 1, 2, 3, 4 또는 5인 정수이고, 상기 X⁴는 산소와 연결된 당류(saccharide)이며; 상기 X² 및 X³은 수소이고; 그리고 상기 X¹은 산소와 연결된 당류일 수 있다.Specifically, the compound of any one of Formulas 1 to 3 is L ¹

N is an integer equal to 1, 2, 3, 4 or 5, and X ⁴ is a saccharide linked to oxygen; X ² and X ³ are hydrogen; And X ¹ may be a saccharide connected with oxygen.

The compound may be one of Formulas 4 to 22 according to an embodiment of the present invention, but is not limited thereto.

Specifically, the extraction may be to extract the membrane protein from the cell membrane.

As used herein, the term "membrane protein" is a generic term for proteins or glycoproteins that are introduced into cell membrane lipid bilayers. It exists in various states such as penetrating the entire layer of the cell membrane, located on the surface layer, or contacting the cell membrane. Examples of membrane proteins include, but are not limited to, enzymes, receptors such as peptide hormones and local hormones, receptors such as sugars, ion channels, and cell membrane antigens.

The membrane protein includes any protein or glycoprotein introduced into the cell membrane lipid bilayer, and specifically, LHI-RC, LeuT (Leucine transporter), β ₂ AR (human β ₂ adrenergic receptor), MelB _st (melibiose permease) or a combination of two or more thereof, but is not limited thereto.

By using a steroid-based compound according to embodiments of the present invention, membrane proteins can be stably stored in an aqueous solution for a long time compared to existing compounds, and can be utilized for functional analysis and structural analysis.

Membrane protein structure and function analysis is one of the fields of greatest interest in current biology and chemistry, and thus it is applicable to the study of protein structure closely related to drug development.

In addition, the compound according to the embodiments of the present invention can be synthesized from a readily available starting material by a simple method, thereby enabling mass production of the compound for membrane protein research.

1 is a diagram showing a synthesis scheme of GDNs according to Embodiment 1 of the present invention.
2 is a diagram showing a synthesis scheme of SPSs and SPS-Ls according to Example 2 of the present invention.
3 is a diagram showing a synthesis scheme of SMAs according to Example 3 of the present invention.
4 shows a DLS profile of GDNs, SPSs or SPS-Ls.
5 shows a DLS profile of GDNs, SPSs or SPS-Ls.
6 shows the DLS profile of GDNs, SPSs or SPS-Ls.
7 shows the DLS profile of SMAs.
FIG. 8 shows the results of measuring the structural stability of LeuT (Leucine transporter) in aqueous solution by GMCs, SPSs, SPS-Ls, or DDM of CMC + 0.04 wt%. Protein stability was confirmed by measuring the substrate binding characteristics of the transporter through a scintillation proximity assay (SPA). LeuT was incubated for 13 days at room temperature in the presence of each amphiphilic compound, and the substrate binding properties of the protein were measured at regular intervals:
Figure 9 is the result of measuring the structural stability of LeuT (Leucine transporter) in the aqueous solution by CMC + 0.04 wt% SMAs or DDM. Protein stability was confirmed by measuring the substrate binding characteristics of the transporter through a scintillation proximity assay (SPA). LeuT was incubated at room temperature for 14 days in the presence of each amphiphilic compound to measure the substrate binding properties of the protein at regular intervals:
Figure 10 is the result of measuring the effect on the stability of β ₂ AR by GDNs, SPSs, SPS-Ls or DDM at regular intervals for 5 days. Protein ligand binding properties were measured by ligand binding assay of [ ³ H] -dihydroalprenolol (DHA).
11 is a result of measuring the effect on the stability of β ₂ AR by GDNs, SPSs, SPS-Ls, or DDM of CMC + 0.2 wt% at an initial 30 minutes. Protein ligand binding properties were measured by ligand binding assay of [ ³ H] -dihydroalprenolol (DHA).
Figure 12 shows that MelB _St protein is extracted at three temperatures (65, 70, 75 ℃) using GDNs, SPSs, SPS-Ls or DDM at a concentration of 1.5 wt%, incubated at the same temperature for 90 minutes, and then in the aqueous solution. The result of measuring the amount of dissolved MelB _St protein is:
(a) SDS-PAGE and Western Blotting results showing the amount of MelB _St protein extracted using each amphipathic compound and the amount of MelB _St protein extracted using each amphiphilic compound were applied to the untreated amphiphilic membrane sample (Memb). Histogram in percent of total protein present; And
(b) Checking whether the function of MelB _St dissolved by DDM, SPS-3 or SPS-3L is maintained.
Figure 13 shows that MelB _St protein is extracted at four temperatures (0, 45, 55 or 65 ° C.) using SMAs or DDM at a concentration of 1.5 wt%, incubated at the same temperature for 90 minutes, and then dissolved in MelBSt. Results of SDS-PAGE and Western Blotting, which measure the amount of protein:
FIG. 14 shows the amount of MelB _St protein dissolved in an aqueous solution after extraction of MelB _St protein at 23 ° C. using GDNs, SPSs, SPS-Ls, or DDM at a concentration of 1.5 wt%, followed by incubation for 90 minutes. It is a result of a measurement

Hereinafter, the present invention will be described in more detail in the following examples. However, the following examples merely illustrate the contents of the present invention and do not limit or limit the scope of the present invention. From the detailed description and examples of the present invention, those skilled in the art to which the present invention pertains can be easily inferred to be within the scope of the present invention.

< Example  1> GDNs  Synthesis method of

<1- 1> dioxenine Hydrogenation

A hydroxyl group was introduced to carbon 2, 3 or 6 of diosgenin.

<1-2> Of maltosylation reactions  General Synthesis Procedure (Step d of Figure 1)

A mixture of the compound produced in Example 1-1, 2,4,5-collidine (1.0 equiv) and AgOTf (2.4 equiv) was stirred at -45 ° C in anhydrous CH ₂ Cl ₂ . A solution of perbenzoylated maltosylbromide / perbenzoylated glucosylbromide in anhydrous CH ₂ Cl ₂ was added dropwise to this suspension for 30 minutes. After stirring was continued at −45 ° C. for 30 minutes, the temperature of the reaction mixture was increased to 0 ° C. and stirring was continued for 1 hour. After completion of the reaction (indicated by TLC), the reaction was quenched by addition of pyridine, then diluted with CH ₂ Cl ₂ and filtered over celite. The filtrate was washed successively with 1 M aqueous Na ₂ S ₂ O ₃ solution (40 mL), 0.1 M aqueous HCl solution (40 mL) and brine (2 × 40 mL). The organic layer was dried over anhydrous Na ₂ SO _4, and then the solvent was removed with a rotary evaporator. The residue obtained was purified by silica gel column chromatography (EtOAc / hexanes) to afford the desired compound as a white solid.

<1-3> Deprotection Vaporization  reaction ( deprotection  General synthetic procedure for the reaction (step e of FIG. 1)

This was followed by the synthetic method of Chae, PS et al. (Nat Meth 2010, 7, 1003.). De-O-benzoylation was performed under Zemplen's conditions. The O-protected compound was dissolved in anhydrous CH ₂ Cl ₂ and then MeOH was added slowly until a continuous precipitation appeared. To the reaction mixture was added 0.5 M of methanolic solution, NaOMe, to a final concentration of 0.05 M. The reaction mixture was stirred at room temperature for 6 hours. After completion of the reaction, the reaction mixture was neutralized with Amberlite IR-120 (H + form) resin. The resin was removed by filtration, washed with MeOH and the solvent was removed from the filtrate in vacuo. The residue was purified by silica gel chromatography (MeOH: CH ₂ Cl ₂ ) to afford the title compound as a white solid.

< Production Example  1> GDN - 1 of  Synthetic Method

The synthesis scheme of GDN-1 is shown in FIG. 1. Compounds of GDN-1 were synthesized according to the synthesis methods of the following <1-1> to <1-5>.

<1-1> Of tycogenin (compound A)  Synthesis (step a of FIG. 1)

Palladium (10%) on activated carbon (500 mg) was added to a solution of diosgenin (5.0 g) mixed in HOAc (250 mL) and the resulting mixture was stirred for 24 h under hydrogen atmosphere (2 bar). The reaction mixture was filtered through celite and the solvent removed by rotary evaporator to give the desired tichogenin (Compound A) in 98% yield.

<1-2> Synthesis of Compound B (Step b of FIG. 1)

Compound B was prepared under the same conditions according to the references (SK Upadhyay, CC Creech, KL Bowdy, ED Stevens, BS Jursic and DM Neumann, Bioorg . Med . Chem . Lett . 2011, 21 , 2826-2831).

<1-3> Synthesis of Compound C (Step c of FIG. 1)

Dichloromethane (10 mL) solution of methanol (10 mL) and Compound B (0.5 g) and sodium hydroxide (2.4 equiv) were stirred overnight at room temperature. Acidic resin-Dowex (˜1 g) was added to the reaction mixture and the suspension was stirred at room temperature for 15 minutes. The resin was separated by filtration and the solvent removed by rotary evaporator to yield compound C in 98% yield.

¹ H NMR (400 MHz, CD ₃ OD): δ 4.38 (q, J = 8.0 Hz, 1H), 3.79 (br s, 1 H), 3.75 (br s, 1 H), 3.45-3.42 (m, 1 H), 3.31 -3.30 (m, 4H), 1.98-1.10 (m, 26H), 1.01 (s, 3H), 0.95 (d, J = 8.0 Hz, 3H), 0.79-0.78 (m, 7H); ¹³ C NMR (100 MHz, CD ₃ OD): δ 110.7, 82.4, 72.3, 71.2, 68.0, 64.0, 57.8, 56.8, 43.1, 41.9, 41.4, 41.0, 40.3, 37.1, 36.1, 33.7, 32.8, 32.6, 31.6, 30.0, 29.5, 22.0, 17.7, 17.2, 15.0, 14.8.

<1-4> Of maltosylation reactions  General Synthesis Procedure (Step d of Figure 1)

GDN-1a was obtained in 83% yield following the general saccharification reaction procedure of Example 1-2.

¹ H NMR (400 MHz, CDCl ₃ ): δ 8.12-7.68 (m, 28H), 7.57-7.20 (m, 42H), 6.15 (t, J = 8.0 Hz, 2H), 5.77-5.63 (m, 6H) , 5.28-5.19 (m, 4H), 4.83-4.29 (m, 14H), 3.79-3.34 (m, 6H), 1.91-1.08 (m, 16H), 0.96 (d, J = 8.0 Hz, 3H), 0.86 -0.83 (m, 6H), 0.77 (d, J = 4.0 Hz, 2H), 0.66 (s, 3H), 0.59 (s, 3H); ¹³ C NMR (100 MHz, CDCl ₃ ): δ 165.9 (2C), 165.8, 165.5, 165.2, 165.1, 165.0 (2C), 133.7, 133.6, 133.5, 133.4, 133.3, 130.1, 130.0, 129.9, 129.8, 129.7, 129.6, 129.4, 129.3 (2C), 129.0, 128.8 (2C), 128.7, 128.6, 128.5, 128.3 (2C), 109.3, 100.5, 99.9, 96.4, 96.3, 80.9, 75.9, 74.9, 74.8, 73.1, 72.8, 72.7 , 72.6, 71.1, 71.0, 70.0, 69.2 (2C), 63.6, 63.4, 62.6, 62.2, 56.1, 54.7, 41.7, 40.5, 39.9, 38.7, 38.5, 35.2, 34.3, 31.9, 31.7, 31.6, 30.4, 29.7, 28.9, 27.6, 20.4, 17.3, 16.5, 14.6, 13.5.

<1-5> Deprotection Vaporization  reaction ( deprotection  General synthetic procedure for the reaction (step c of FIG. 1)

Compound GDN-1 was obtained in 94% yield following the general synthetic procedure for deprotection reaction of Examples 1-3.

¹ H NMR (400 MHz, CD ₃ OD): δ 5.17 (d, J = 4.0 Hz, 2H), 4.38-4.34 (m, 2H), 4.30 (d, J = 8.0 Hz, 1H), 3.96 (br s , 1H), 3.92-3.71 (m, 7H), 3.67-3.16 (m, 24H), 1.97-1.86 (m, 4H), 1.73-1.10 (m, 20H), 0.98 (s, 3H), 0.95 (d , J = 8.0 Hz, 3H), 0.78 (s, 6H); ¹³ C NMR (100 MHz, CD ₃ OD): δ 110.7, 103.8, 103.7, 103.0, 98.3, 82.4, 81.5, 81.4, 78.2, 78.1, 76.8, 76.7, 76.3, 75.2, 74.9 (2C), 74.8, 74.3, 71.6, 68.0, 64.0, 62.9, 62.4, 57.9, 56.4, 55.0, 43.0, 41.9, 41.4, 40.6, 40.3, 36.8, 36.2, 33.6, 32.8, 32.6, 31.7, 31.6, 30.0, 29.4, 21.9, 17.7, 17.2, 15.0, 14.4; HRMS ( FAB ⁺ ) : calcd. for C ₅₁ H ₈₄ O ₂₄ [M + Na] ⁺ 1081.5431, observed 1081.5428.

< Production Example  2> GDN Synthesis of -2

The synthesis scheme of GDN-2 is shown in FIG. 1. Compounds of GDN-2 were synthesized according to the synthesis methods of <2-1> to <2-3>.

<2-1> Synthesis of Compound E

Compound E was prepared from ticogenin under the same conditions according to the references ( SK Upadhyay, CC Creech, KL Bowdy, ED Stevens, BS Jursic and DM Neumann, Bioorg . Med . Chem . Lett. 2011, 21 , 2826-2831.) It was.

¹ H NMR (400 MHz, CDCl ₃ ): δ 4.38 (q, J = 8.2 Hz, 1H), 3.95 (br s, 1 H), 3.77 (br s, 1 H), 3.49-3.45 (m, 1 H), 3.37 (t, J = 10.8 Hz, 1H), 2.17 (br s, 1H), 1.98-1.09 (m, 33H), 0.95 (d, J = 7.2 Hz, 3H), 0.81 (s, 3H), 0.78 (d , J = 6.4 Hz, 3H), 0.76 (s, 3H); ¹³ C NMR (100 MHz, CDCl ₃ ): δ 119.5, 81.0, 69.4, 69.3, 67.0, 62.3, 56.4, 54.4, 41.8, 41.1, 40.8, 40.2, 38.3, 37.2, 34.6, 34.5, 32.2, 32.0, 31.6, 30.5, 29.9, 29.0, 27.8, 20.9, 17.4, 16.7, 14.7, 12.7.

<2-2> GDN Synthesis of -2a

Compound GND-2a was synthesized in a yield of 87% according to the general saccharification reaction procedure of Example 1-2. ¹ H NMR (400 MHz, CDCl ₃ ): δ 8.13-7.68 (m, 27H), 7.57-7.19 (m, 44H), 6.94 (t, J = 8.0 Hz, 2H), 6.77-6.71 (m, 2H) , 6.18-5.66 (m, 8H), 5.40-5.31 (m, 4H), 4.92-3.34 (m, 19H), 1.90-1.08 (m, 34H), 0.87 (d, J = 8.0 Hz, 3H), 0.90 -0.80 (m, 10H), 0.77 (d, J = 8.0 Hz, 3H), 0.61 (s, 3H), 0.53 (s, 3H); ¹³ C NMR (100 MHz, CDCl ₃ ): δ 166.4, 165.9 (2C), 165.8, 165.5, 165.2, 165.1, 164.8, 133.7, 133.5, 133.4, 133.3, 130.1, 130.0, 129.9, 129.7, 129.6, 129.5 (2C ), 129.3, 129.0, 128.8 (2C), 128.7, 128.6, 128.2 (2C), 109.3, 98.7, 98.2, 96.6, 95.5, 75.7, 74.8, 74.8, 73.1, 72.8, 72.7, 72.2, 71.1, 71.0, 70.5, 70.0, 69.3, 68.9, 63.8, 62.7, 62.2, 55.4, 53.1, 41.7, 40.4, 39.2, 38.6, 38.5, 34.1, 32.9, 32.1, 31.8, 31.5, 31.6, 30.4, 30.2, 29.8, 28.9, 27.2, 22.8, 21.2, 20.4, 19.9, 17.3, 16.5, 14.6, 14.3, 12.8.

<2-3> GDN Synthesis of -2

GDN-2 in 95% yield according to the general synthetic procedure for deprotection reaction of Examples 1-3. Synthesized. ¹ H NMR (400 MHz, DMSO-d ₆ ): δ 5.54 (d, J = 4.0 Hz, 4H), 4.99-4.97 (m, 7H), 4.57-4.33 (m, 8H), 4.03 (br s, 1H ), 3.73-3.04 (m, 31H), 3.02-3.00 (m, 4H), 2.50 (br s, 1H), 1.89-1.11 (m, 24H), 0.75 (d, J = 8.0 Hz, 3H), 0.72 -0.68 (m, 10 H); ¹³ C NMR (100 MHz, DMSO-d ₆ ): δ 118.4, 103.5, 100.9, 80.2, 79.9, 79.7, 76.9, 76.4, 76.2, 76.1, 75.1, 73.7, 73.5, 73.3, 72.5, 69.9, 65.9, 61.9, 60.8, 60.7, 55.6, 53.7, 41.1, 36.3, 34.0, 31.8, 30.9, 29.8, 28.5, 27.1, 17.1, 16.3, 14.7; HRMS ( FAB ⁺ ) : calcd. for C ₅₁ H ₈₄ O ₂₄ [M + Na] ⁺ 1081.5431, observed 1081.5428.

< Production Example  3> GDN Synthesis of -3

The synthesis scheme of GDN-3 is shown in FIG. 1. Compounds of GDN-2 were synthesized according to the synthesis methods of the following <3-1> to <1-5>.

<3-1> Synthesis of Compound F

1M BH ₃ (3.6 mL, 3.6 mmol) mixed in THF was slowly added to a solution of diosgenin (500 mg, 1.2 mmol) mixed in dry THF (50 mL) at 0 ° C. under argon atmosphere. The mixture was stirred at rt overnight. NaOH (10 N, 8 mmol) was added at 0 ° C over 45 minutes. 30% hydrogen peroxide (0.75 mL, 6.5 mmol) was then added and vigorous stirring was continued for 3 hours at room temperature. Extracted with CH ₂ Cl ₂ (120 mL) and the combined extracts were washed successively with 1N HCl, saturated NaHCO ₃ and brine, dried (anhydrous Na ₂ SO ₄ ), filtered and concentrated in vacuo. Purification by column chromatography gave the compound F as a white solid in 93% yield.

<3-2> GDN Synthesis of -3a

Compound GDN-3a was synthesized in a yield of 88% according to the general saccharification reaction procedure of Example 1-2. ¹ H NMR (400 MHz, CDCl ₃ ): δ 8.10-7.74 (m, 28H), 7.57-7.03 (m, 44H), 7.05 (t, J = 8.0 Hz, 2H), 6.77-6.12 (m, 2H) , 5.94-5.81 (m, 6H), 5.37-5.12 (m, 4H), 4.94-3.19 (m, 18H), 2.08-1.08 (m, 34H), 0.87 (d, J = 8.0 Hz, 3H), 0.90 -0.80 (m, 10H), 0.77 (d, J = 8.0 Hz, 3H), 0.68 (s, 3H), 0.53 (s, 3H); ¹³ C NMR (100 MHz, CDCl ₃ ): δ 167.0, 166.1, 165.9 (2C), 165.4, 165.2, 164.7, 133.8, 133.5, 133.4 (2C), 133.3, 130.1, 130.0 (2C), 129.8, 129.6, 129.5 , 129.3 (2C), 129.0, 128.8 (2C), 128.6 (2C), 128.2 (2C), 109.3, 99.1, 98.9, 97.6, 95.5, 75.7, 74.8, 74.8, 74.1, 72.8, 72.7, 72.2, 71.1, 71.0 (2C), 69.3, 68.7, 63.5, 62.2, 62.1, 55.4, 53.0, 41.9, 40.0, 39.2, 38.6, 38.5, 38.4, 34.1, 32.9, 32.2, 31.8, 31.7, 31.5, 31.6, 30.4, 30.2, 29.9, 27.2, 22.8, 21.3, 20.0, 19.9, 17.4, 16.4, 14.6 (2C), 12.8.

<3-3> GDN Synthesis of -3

According to the general synthetic procedure for the deprotection reaction of Example 1-3, GDN-3 was obtained in a yield of 96%. Synthesized. ¹ H NMR (400 MHz, CD ₃ OD): δ 5.16 (d, J = 4.0 Hz, 2H), 4.49 (d, J = 8.0 Hz, 1H), 4.39-4.34 (m, 1H), 4.32 (d, J = 8.0 Hz, 1H), 3.87-3.19 (m, 34H), 2.61 (br s, 1H), 2.23-2.20 (m, 1H), 2.02-1.41 (m, 28H), 0.95 (d, J = 8.0 Hz, 3H), 0.90 (s, 3H), 0.80 (s, 3H); ¹³ C NMR (100 MHz, CD ₃ OD): δ 110.6, 103.0, 102.8, 102.3, 98.3, 93.9, 82.4, 82.3, 81.9, 81.5, 81.4, 79.9, 78.2, 78.1, 77.9, 76.8, 75.2, 75.1, 75.0 , 74.8, 74.4, 74.2, 73.5, 72.5, 71.8, 71.7, 71.6, 68.0, 62.8, 62.3, 57.6, 57.4, 55.8, 55.0, 43.0, 42.0, 41.9, 41.7, 40.9, 40.0, 37.0, 36.0, 33.0, 32.5 , 31.7, 31.5, 31.4, 30.6, 30.0, 26.5, 22.1, 17.7, 17.2, 16.3, 15.1; HRMS ( FAB ⁺ ) : calcd. for C ₅₁ H ₈₄ O ₂₄ [M + Na] ⁺ 1081.5431, observed 1081.5428.

< Example  2> SPSs  or SPS - Ls  Synthetic Method

The synthesis scheme of SPSs or SPS-Ls is shown in FIG. 2. According to the synthesis method of <2-1> to <2-2>, three compounds of SPSs and three compounds of SPS-Ls were synthesized.

<2-1> Saccharification  Synthesis of Reaction Substrate (Compounds B1-B4)

0 ° C. in cholestanol (compound A1), cholesterol (compound A2), cytosterol (compound A3) or diosgenin (compound 4) (38 mmol) mixed in dry pyridine (15 mL) and dry CHCl ₃ (15 mL) Treated with p-toluenesulfonylchloride (54 mmol). A catalytic amount of DMAP was also added. The reaction mixture was then stirred for 12 hours. The reaction mixture was washed sequentially with dilute HCl aqueous solution, saturated brine solution and water. The organic phase was dried over anhydrous Na ₂ SO ₄ . The solvent was removed with a vacuum evaporator and recrystallized with chloroform and methanol. The residue (13 mmol) was dissolved in anhydrous dioxane (20 mL) and 1,3-propane diol (0.32 mol) was added. The mixture was refluxed at 110 ° C for 4 h. The solution was cooled and the solvent removed in vacuo. After dissolving the white residue in CH ₂ Cl ₂ , the organic layer was washed with NaHCO ₃ , water and brine and dried over anhydrous Na ₂ SO ₄ . Finally, the product was purified by silica gel column chromatography extracting with n-hexane / ethyl acetate to give compound B1, B2, B3 or B4.

<2-2> Saccharification  reaction ( SPSs  or SPS - Ls  synthesis)

SPSs or SPS-Ls were synthesized from the compound A1-A4 or the compound B1-B4 through general glycosylation reaction (Example 1-2) and deprotection group reaction (Example 1-3).

< Production Example  4> SPS Synthesis of -1

<4-1> Synthesis of Compound 1

Compound 1 was obtained in 88% yield from compound A1 following the general saccharification and deprotection reactions of examples 1-2 and 1-3. ¹ H NMR (400 MHz, (CD ₃ ) ₂ SO): δ 4.89 (d, J = 5.3 Hz, 1H), 4.82 (d, J = 3.7 Hz, 1H), 4.75 (d, J = 4.7 Hz, 1H ), 4.52-4.40 (m, 2H), 3.66 (d, J = 9.4 Hz, 1H), 3.50-3.41 (m, 3H), 3.19-3.11 (m, 1H), 3.10-3.00 (m, 1H), 2.41-2.18 (m, 2H), 2.02-1.77 (m, 5H), 1.61-0.80 (m, 34H), 0.70 (s, 3H); ¹³ C NMR (100 MHz, (CD ₃ ) ₂ SO): δ 97.0, 76.4, 73.2, 72.9, 71.8, 70.3, 61.0, 56.2, 55.7, 49.5, 41.8, 40.2, 39.9, 39.8, 36.6, 36.3, 35.6, 35.2, 31.4, 31.3, 27.7, 27.4, 27.4, 23.9, 23.2, 22.6, 22.4, 20.5, 19.1, 18.5, 11.9.

<4-2> SPS Synthesis of -1a

SPS-1a was obtained in a yield of 75% according to the general saccharification reaction of Example 1-2 from compound 1. ¹ H NMR (400 MHz, CDCl ₃ ): δ 8.25 (d, J = 8.0 Hz, 2H), 8.17-7.78 (m, 24H), 7.79-7.67 (m, 5H), 7.66-7.59 (m, 3H) , 7.58-7.12 (m, 46H), 5.96 (t, J = 8.0 Hz, 1H), 5.93-5.81 (m, 3H), 5.71-5.69 (m, 2H), 5.61-5.42 (m, 6H), 5.01 (d, J = 8.0 Hz, 1H), 4.98-4.92 (m, 2H), 4.88-4.79 (m, 2H), 4.68-4.4.35 (m, 12H), 4.22-4.03 (m, 4H), 3.88 -3.85 (m, 2H), 3.39-3.20 (m, 10H), 2.85 (br s, 1H), 2.75 (br s, 1H), 1.96 (d, J = 12.0 Hz, 1H), 1.90-1.01 (m , 40H), 0.99 (s, 3H), 0.91 (d, J = 8.0 Hz, 3H), 0.88-0.84 (m, 6H), 0.68 (s, 3H); ¹³ C NMR (100 MHz, CDCl ₃ ): δ 166.2, 166.1, 165.9, 165.5, 165.3, 165.2, 165.1, 164.9, 133.7, 133.3, 133.2, 130.2, 130.1, 129.9, 129.6, 129.4, 129.1, 128.9, 128.6, 128.5, 128.4, 128.3, 100.1, 100.0, 99.9, 99.8, 98.6, 78.9, 73.4, 73.0, 71.9, 69.7, 65.3, 56.9, 56.3, 50.1, 42.5, 39.9, 39.7, 39.4, 37.4, 37.0, 36.4, 35.9, 32.1, 31.9, 29.9, 28.4, 28.2, 24.4, 23.9, 23.0, 22.7, 21.2, 19.5, 18.9, 12.4, 12.2, 11.6.

<4-3> SPS Synthesis of -1

Following the general synthetic procedure for the deprotection reaction of Examples 1-3, SPS-1 was obtained in a yield of 90%. Synthesized. ¹ H NMR (400 MHz, CD ₃ OD): δ 5.20 (d, J = 4.0 Hz, 1H), 4.97 (d, J = 8.0 Hz, 1H), 4.63 (d, J = 8.0 Hz, 1H), 4.53 (d, J = 8.0 Hz, 1H), 4.33 (d, J = 8.0 Hz, 1H), 4.20 (q, J = 12.0 Hz, 2H), 4.04-4.01 (m, 1H), 3.93-3.80 (m, 6H), 3.70-3.54 (m, 6H), 3.50-3.14 (m, 26H), 1.97 (d, J = 12.0 Hz, 2H), 1.82-1.75 (m, 1H), 1.71-1.47 (m, 8H) , 1.37-1.20 (m, 14 H), 1.13-0.80 (m, 26 H), 0.65 (s, 3 H); ¹³ C NMR (100 MHz, CD ₃ OD): δ 105.4, 104.8, 103.3, 102.4, 98.7, 82.7, 79.8, 79.0, 78.5, 78.3, 78.2, 78.0, 77.8, 77.7, 77.1, 75.8, 75.5, 75.2, 75.1 , 72.0, 71.8, 71.5, 63.1, 63.0, 62.7, 58.3, 57.7, 55.9, 51.8, 50.0, 43.6, 41.3, 40.8, 38.5, 37.9, 37.5, 37.3, 33.4, 31.2, 29.5, 29.3, 29.0, 25.5, 25.0 , 23.3, 23.1, 22.5, 20.0, 19.4, 12.9, 12.6; HRMS ( FAB ⁺ ) : calcd. for C ₅₇ H ₉₈ O ₂₆ [M + Na] ⁺ 1221.6244, observed 1221.6247.

< Production Example  5> SPS Synthesis of -2

<5-1> Synthesis of Compound 2

Compound 2 was obtained in 90% yield from compound A2 following the general saccharification and deprotection reactions of Examples 1-2 and 1-3. ¹ H NMR (400 MHz, (CD ₃ ) ₂ SO): δ 5.36 (d, J = 4.5 Hz, 1H), 4.90 (d, J = 5.3 Hz, 1H), 4.82 (d, J = 3.7 Hz, 1H ), 4.75 (d, J = 4.7 Hz, 1H), 4.52-4.40 (m, 2H), 3.66 (d, J = 9.4 Hz, 1H), 3.50-3.41 (m, 3H), 3.19-3.11 (m, 1H), 3.09-3.00 (m, 1H), 2.41-2.18 (m, 2H), 2.02-1.77 (m, 5H), 1.61-0.80 (m, 34H), 0.69 (s, 3H); ¹³ C NMR (100 MHz (CD ₃ ) ₂ SO): δ 140.7, 121.1, 96.9, 76.4, 73.2, 72.8, 71.8, 70.4, 61.0, 56.2, 55.6, 49.5, 41.8, 40.2, 39.9, 39.8, 36.6, 36.2 , 35.6, 35.2, 31.4, 31.3, 27.7, 27.4, 27.4, 23.9, 23.2, 22.6, 22.4, 20.6, 19.1, 18.5, 11.7.

<5-2> SPS Synthesis of -2a

SPS-2a was obtained in 76% yield according to the general saccharification reaction of Example 1-2 from compound 2. ¹ H NMR (400 MHz, CDCl ₃ ): δ 8.25 (d, J = 8.0 Hz, 2H), 8.17-7.78 (m, 24H), 7.79-7.67 (m, 5H), 7.66-7.59 (m, 3H) , 7.58-7.12 (m, 46H), 5.96 (t, J = 8.0 Hz, 1H), 5.93-5.81 (m, 3H), 5.71-5.69 (m, 2H), 5.61-5.42 (m, 8H), 5.01 (d, J = 8.0 Hz, 1H), 4.93-4.92 (m, 2H), 4.88-4.79 (m, 2H), 4.67-4.4.35 (m, 12H), 4.22-4.03 (m, 4H), 3.88 -3.85 (m, 2H), 3.39-3.20 (m, 10H), 2.85 (br s, 1H), 2.75 (br s, 1H), 1.96 (d, J = 12.0 Hz, 1H), 1.90-1.01 (m , 40H), 0.99 (s, 3H), 0.91 (d, J = 8.0 Hz, 3H), 0.88-0.84 (m, 6H), 0.67 (s, 3H); ¹³ C NMR (100 MHz, CDCl ₃ ): δ 166.2, 166.1, 165.9, 165.8, 165.4, 165.3, 165.1, 165.0, 164.8, 164.5, 140.7, 133.5, 133.3, 133.2, 130.1, 130.0, 129.9, 129.8, 129.7, 129.6, 129.4, 129.0, 128.6, 128.5, 128.4, 128.3, 121.6, 100.1, 100.0, 99.9, 99.8, 98.6, 78.9, 73.4, 73.0, 71.9, 69.7, 65.3, 56.9, 56.3, 50.1, 42.5, 39.9, 39.7, 39.4, 37.4, 37.0, 36.4, 35.9, 32.1, 31.9, 28.4, 28.2, 24.4, 23.9, 23.0, 22.7, 21.2, 19.5, 18.9, 12.0.

<5-3> SPS Synthesis of -2

According to the general synthetic procedure for the deprotection reaction of Example 1-3, SPS-2 was obtained at a yield of 90%. Synthesized. ¹ H NMR (400 MHz, CD ₃ OD): δ 5.34 (d, J = 4.0 Hz, 1H), 5.21 (d, J = 4.0 Hz, 1H), 4.98 (d, J = 8.0 Hz, 1H), 4.65 (d, J = 8.0 Hz, 1H), 4.55 (d, J = 8.0 Hz, 1H), 4.33 (d, J = 8.0 Hz, 1H), 4.22 (q, J = 12.0 Hz, 2H), 4.05-4.02 (m, 1H), 3.96-3.80 (m, 6H), 3.71-3.55 (m, 6H), 3.45-3.16 (m, 26H), 2.33-2.30 (m, 2H), 2.03-1.83 (m, 6H) , 1.59-1.25 (m, 12H), 1.16-0.80 (m, 24H), 0.69 (s, 3H); ¹³ C NMR (100 MHz, CD ₃ OD): δ 142.2, 122.9, 105.4, 104.7, 103.3, 102.4, 98.7, 82.7, 79.8, 79.0, 78.5, 78.3, 78.2, 78.0, 77.8, 77.7, 77.1, 75.8, 75.5 , 75.2, 75.1, 72.0, 71.8, 71.5, 63.1, 63.0, 62.7, 58.3, 57.7, 51.8, 50.0, 43.6, 41.3, 40.8, 38.5, 37.9, 37.5, 37.3, 33.4, 31.2, 29.5, 29.3, 29.0, 25.5 , 25.0, 23.3, 23.1, 22.4, 20.0, 19.4, 12.9, 12.5; HRMS ( FAB ⁺ ) : calcd. for C ₅₇ H ₉₆ O ₂₆ [M + Na] ⁺ 1219.6080, observed 1219.6085.

< Production Example  6> SPS Synthesis of -3

<6-1> Synthesis of Compound 3

Compound 3 was obtained in a yield of 88% according to the general saccharification and deprotection reaction of Examples 1-2 and 1-3 from Compound A3. ¹ H NMR (400 MHz, (CD ₃ ) ₂ SO): δ 5.36 (d, J = 4.5 Hz, 1H), 4.91 (d, J = 5.3 Hz, 1H), 4.83 (d, J = 3.7 Hz, 1H ), 4.76 (d, J = 4.7 Hz, 1H), 4.54-4.40 (m, 2H), 3.66 (d, J = 9.4 Hz, 1H), 3.50-3.41 (m, 3H), 3.20-3.11 (m, 1H), 3.09-3.00 (m, 1H), 2.41-2.18 (m, 2H), 2.02-1.77 (m, 5H), 1.61-0.80 (m, 34H), 0.68 (s, 3H); ¹³ C NMR (100 MHz, (CD ₃ ) ₂ SO): δ 141.5, 121.3, 96.9, 76.4, 73.2, 72.8, 71.8, 70.4, 61.0, 56.2, 55.6, 49.5, 41.8, 40.2, 39.9, 39.8, 36.6, 36.2, 35.6, 35.2, 31.4, 31.3, 29.9, 29.7, 27.7, 27.4, 27.4, 23.9, 23.2, 22.6, 22.4, 20.6, 19.1, 18.5, 14.1, 11.7.

<6-2> SPS Synthesis of -3a

SPS-3a was obtained in 78% yield according to the general saccharification reaction of Example 1-2 from compound 3. ¹ H NMR (400 MHz, CDCl ₃ ): δ 8.25 (d, J = 8.0 Hz, 2H), 8.17-7.78 (m, 24H), 7.79-7.67 (m, 5H), 7.66-7.59 (m, 3H) , 7.58-7.12 (m, 46H), 5.97 (t, J = 8.0 Hz, 1H), 5.93-5.81 (m, 3H), 5.71-5.69 (m, 2H), 5.61-5.42 (m, 8H), 5.01 (d, J = 8.0 Hz, 1H), 4.93-4.92 (m, 2H), 4.88-4.79 (m, 2H), 4.67-4.4.35 (m, 12H), 4.22-4.03 (m, 6H), 3.88 -3.85 (m, 2H), 3.39-3.20 (m, 10H), 2.85 (br s, 1H), 2.75 (br s, 1H), 1.96 (d, J = 12.0 Hz, 1H), 1.90-1.01 (m , 45H), 0.99 (s, 3H), 0.91 (d, J = 8.0 Hz, 3H), 0.88-0.84 (m, 6H), 0.69 (s, 3H); ¹³ C NMR (100 MHz, CDCl ₃ ): δ 166.1, 165.9, 165.4, 165.3, 165.2, 165.1, 165.0, 164.7, 164.4, 140.7, 133.6, 133.2, 130.2, 129.9, 129.7, 129.6, 129.5, 129.3, 129.1, 128.7, 128.6, 128.5, 128.4, 128.3, 121.6, 100.1, 100.0, 99.9, 99.8, 98.6, 78.9, 73.4, 73.0, 72.0, 69.9, 63.7, 57.0, 56.2, 45.9, 42.5, 39.9, 36.7, 36.4, 34.1, 32.0, 31.9, 29.3, 28.5, 27.9, 26.2, 24.5, 23.2, 21.2, 20.0, 19.5, 19.2, 19.0, 18.9, 12.0, 11.5.

<6-3> SPS Synthesis of -3

Following the general synthetic procedure for the deprotection reaction of Examples 1-3, SPS-3 was obtained in a yield of 91%. Synthesized. ¹ H NMR (400 MHz, CD ₃ OD): δ 5.33 (d, J = 4.0 Hz, 1H), 5.20 (d, J = 4.0 Hz, 1H), 4.97 (d, J = 8.0 Hz, 1H), 4.64 (d, J = 8.0 Hz, 1H), 4.54 (d, J = 8.0 Hz, 1H), 4.32 (d, J = 8.0 Hz, 1H), 4.20 (q, J = 12.0 Hz, 2H), 4.04-4.01 (m, 1H), 3.96-3.80 (m, 6H), 3.70-3.54 (m, 6H), 3.44-3.14 (m, 26H), 2.32-2.30 (m, 2H), 2.03-1.83 (m, 6H) , 1.59-1.25 (m, 15H), 1.16-0.80 (m, 26H), 0.68 (s, 3H); ¹³ C NMR (100 MHz, CD ₃ OD): δ 142.2, 122.9, 105.4, 104.7, 103.3, 102.4, 98.7, 82.7, 79.8, 79.0, 78.5, 78.3, 78.2, 78.0, 77.8, 77.7, 77.1, 75.8, 75.5 , 75.2, 75.1, 72.0, 71.8, 71.5, 63.1, 63.0, 62.7, 58.3, 57.7, 51.8, 50.0, 47.3, 43.6, 41.3, 40.8, 38.5, 37.9, 37.5, 37.3, 35.2, 33.3, 31.2, 30.5, 29.5 , 29.2, 29.0, 27.2, 25.5, 25.0, 23.3, 23.1, 22.4, 20.3, 19.5, 12.9, 12.5; HRMS ( FAB ⁺ ) : calcd. for C ₅₉ H ₁₀₀ O ₂₆ [M + Na] ⁺ 1247.6401, observed 1247.6406.

< Production Example  7> SPS Synthesis of -4

<7-1> Synthesis of Compound 4

Compound 4 was obtained in 90% yield from compound A4 following the general saccharification and deprotection reactions of Examples 1-2 and 1-3. ¹ H NMR (400 MHz, CDCl ₃ / CD ₃ OD): δ 5.37 (d, J = 3.1 Hz, 1H), 4.42 (q, J = 7.4 Hz, 1H), 4.40 (d, J = 7.8 Hz, 1H ), 3.84 (d, J = 12.0 Hz, 1H), 3.83 (d, J = 4.7 Hz, 1H), 3.58 (m, 1H), 3.47 (d, J = 12.0 Hz, 1H), 3.45-3.20 (m , 5H), 2.41 (d, J = 13.2 Hz, 1 H), 2.27 (m, 1H), 2.05-0.92 (m, 23H), 1.03 (s, 3H), 0.97 (d, J = 6.9 Hz, 3H ), 0.81 (d, J = 6.3 Hz, 3H), 0.80 (s, 3H); ¹³ C NMR (100 MHz, CDCl ₃ / CD ₃ OD): δ 141 .7, 123.0, 110.9, 102.5, 82.3, 80.3, 77.8, 77.2, 74.9, 71 .6, 68.2, 63.3, 63.1, 57.8, 51. 4, 42.9, 41.6, 41 .0, 40.0, 38.5, 38.1, 33.3, 33.0, 32.7, 32.6, 31 .5, 30.8, 30.0, 22.1, 20.5, 18.2, 17.5, 15.6.

<7-2> SPS Synthesis of -4a

SPS-4a was obtained in 75% yield according to the general saccharification reaction of Example 1-2 from compound 4. ¹ H NMR (400 MHz, CDCl ₃ ): δ 8.25 (d, J = 8.0 Hz, 2H), 8.16-7.77 (m, 24H), 7.72-7.67 (m, 5H), 7.66-7.59 (m, 3H) , 7.58-7.15 (m, 46H), 5.96 (t, J = 8.0 Hz, 1H), 5.90-5.82 (m, 3H), 5.71-5.69 (m, 3H), 5.61-5.42 (m, 8H), 5.01 (d, J = 8.0 Hz, 1H), 4.98-4.92 (m, 2H), 4.88-4.79 (m, 2H), 4.69-4.4.35 (m, 12H), 4.22-4.03 (m, 4H), 3.87 -3.85 (m, 2H), 3.40-3.18 (m, 12H), 2.85 (br s, 1H), 2.75 (br s, 1H), 1.96 (d, J = 12.0 Hz, 1H), 1.90-1.01 (m , 42H), 0.99 (s, 3H), 0.91 (d, J = 8.0 Hz, 3H), 0.88-0.84 (m, 6H), 0.70 (s, 6H); ¹³ C NMR (100 MHz, CDCl ₃ ): δ 166.1, 166.0, 165.7, 165.9, 165.8, 165.7, 165.4, 165.3, 165.2, 165.1, 165.0, 164.8, 164.3, 140.7, 133.6, 133.4, 133.3, 133.2, 133.1, 130.2, 130.1, 130.0, 129.8, 129.7, 129.6, 129.5, 129.3, 129.0, 128.8, 128.7, 128.6, 128.5, 128.4, 128.3, 128.2, 121.3, 109.4, 101.5, 100.4, 100.3, 100.2, 99.9, 80.9, 73.1, 72.4, 72.2, 72.0, 69.8, 66.9, 63.5, 62.2, 56.7, 50.0, 41.7, 40.4, 39.9, 36.8, 31.5, 31.4, 30.4, 28.9, 20.9, 19.5, 17.3, 16.4, 14.7.

<7-3> SPS Synthesis of -4

According to the general synthetic procedure for the deprotection reaction of Examples 1-3, SPS-4 was obtained at a yield of 90%. Synthesized. ¹ H NMR (400 MHz, CD ₃ OD): δ 5.33 (d, J = 4.0 Hz, 1H), 5.21 (d, J = 4.0 Hz, 1H), 4.99 (d, J = 8.0 Hz, 1H), 4.66 (d, J = 8.0 Hz, 1H), 4.56 (d, J = 8.0 Hz, 1H), 4.36 (t, J = 8.0 Hz, 2H), 4.22 (q, J = 12.0 Hz, 2H), 4.04-4.01 (m, 1H), 3.96-3.80 (m, 6H), 3.70-3.54 (m, 6H), 3.44-3.14 (m, 26H), 2.36 (d, J = 12.0 Hz, 2H), 2.01-1.80 (m , 6H), 1.75-1.39 (m, 12H), 1.28-0.93 (m, 12H), 0.79 (s, 6H); ¹³ C NMR (100 MHz, CD ₃ OD): δ 142.2, 122.7, 110.7, 105.3, 104.7, 103.2, 102.4, 98.6, 82.6, 82.3, 79.7, 78.4, 78.1, 77.9, 77.8, 77.6, 77.0, 75.7, 75.5 , 75.2, 75.0, 71.9, 71.7, 71.3, 67.9, 63.8, 63.1, 62.9, 62.7, 62.6, 57.9, 51.7, 50.0, 43.0, 41.5, 41.3, 41.0, 38.4, 38.1, 33.3, 32.9, 32.5, 31.5, 30.0 , 28.9, 22.1, 20.1, 17.6, 16.9, 15.1; HRMS ( FAB ⁺ ) : calcd. for C ₅₇ H ₉₂ O ₂₈ [M + Na] ⁺ 1247.5673, observed 1247.5670.

< Production Example  8> SPS Synthesis of -1L

<8-1> Synthesis of Compound 1L

Compound L was obtained in 90% yield from compound B1 following the general saccharification and deprotection reactions of Examples 1-2 and 1-3. ¹ H NMR (400 MHz, CD ₃ OD): δ 4.26 (d, J = 8.0 Hz, 1H), 4.01-3.96 (m, 1H), 3.87-3.84 (d, J = 12.0 Hz, 1H), 3.70- 3.64 (m, 2H), 3.53-3.49 (m, 3H), 3.37-3.12 (m, 6H), 2.01 (d, J = 12.0 Hz, 1H), 1.88-0.82 (m, 42H), 0.69 (s, 3H); ¹³ C NMR (100 MHz, CD ₃ OD): δ 104.7. 78.2, 78.0, 75.5, 75.2, 71.7, 68.3, 65.9, 62.9, 58.1, 57.8, 56.1, 43.9, 41.6, 41.0, 40.8, 37.5, 37.3, 37.1, 37.0, 34.1, 33.5, 31.7, 29.9, 29.5, 29.3, 27.0, 25.4, 25.1, 23.4, 23.2, 22.1, 19.4, 12.8, 12.1.

<8-2> SPS - 1La's  synthesis

From the compound 1L, SPS-1La was obtained in a yield of 80% according to the general saccharification reaction of Example 1-2. ¹ H NMR (400 MHz, CDCl ₃ ): δ 8.24 (d, J = 8.0 Hz, 2H), 8.15-7.78 (m, 24H), 7.72-7.67 (m, 5H), 7.66-7.59 (m, 3H) , 7.58-7.12 (m, 46H), 5.96 (t, J = 8.0 Hz, 1H), 5.92-5.82 (m, 3H), 5.71-5.69 (m, 2H), 5.61-5.42 (m, 6H), 5.01 (d, J = 8.0 Hz, 1H), 4.98-4.92 (m, 2H), 4.88-4.79 (m, 2H), 4.68-4.4.35 (m, 12H), 4.22-4.03 (m, 4H), 3.88 -3.85 (m, 2H), 3.37-3.18 (m, 14H), 2.85 (br s, 1H), 2.75 (br s, 1H), 1.96 (d, J = 12.0 Hz, 1H), 1.90-1.01 (m , 42H), 0.99 (s, 3H), 0.91 (d, J = 8.0 Hz, 3H), 0.88-0.84 (m, 6H), 0.67 (s, 3H); ¹³ C NMR (100 MHz, CDCl ₃ ): δ 166.2, 166.1, 165.9, 165.8, 165.3, 165.2, 165.0, 164.8, 164.5, 133.5, 133.3, 133.2, 130.1, 130.0, 129.9, 129.8, 129.7, 129.6, 129.4, 129.0, 128.6, 128.5, 128.4, 128.3, 100.1, 100.0, 99.9, 99.8, 98.6, 78.9, 73.4, 73.0, 71.9, 69.7, 65.3, 56.9, 56.3, 50.1, 42.5, 39.9, 39.7, 39.4, 37.4, 37.0, 36.4, 35.9, 32.1, 32.0, 29.9, 28.6, 28.4, 28.2, 24.4, 23.9, 23.0, 22.7, 21.2, 19.5, 18.9, 12.0, 11.6.

<8-3> SPS Synthesis of -1L

Following the general synthetic procedure for the deprotection reaction of Examples 1-3, SPS-1L was obtained in 92% yield. Synthesized. ¹ H NMR (400 MHz, CD ₃ OD): δ 5.06 (d, J = 4.0 Hz, 1H), 5.01 (d, J = 8.0 Hz, 1H), 4.67 (d, J = 8.0 Hz, 1H), 4.57 (d, J = 8.0 Hz, 1H), 4.37 (d, J = 8.0 Hz, 1H), 4.25 (d, J = 12.0 Hz, 1H), 3.98-3.39 (m, 3H), 3.88-3.83 (m, 6H), 3.71-3.63 (m, 6H), 3.55-3.47 (m, 4H), 3.38-3.18 (m, 28H), 2.01 (d, J = 12.0 Hz, 1H), 1.88-1.76 (m, 4H) , 1.70-1.02 (m, 26H), 0.94-0.86 (m, 10H), 0.82 (s, 3H), 0.68 (s, 3H); ¹³ C NMR (100 MHz, CD ₃ OD): δ 105.4, 104.8, 103.3, 102.5, 100.1, 82.3, 78.5, 78.2, 78.1, 77.8, 77.7, 75.6, 75.2, 72.0, 71.7, 71.5, 67.0, 66.3, 66.0 , 63.0, 62.6, 58.1, 57.8, 56.0, 43.9, 41.6, 41.0, 40.8, 37.5, 37.3, 37.1, 37.0, 34.2, 33.5, 32.9, 31.2, 29.9, 29.5, 29.3, 27.0, 25.4, 25.1, 23.9, 23.4 , 23.1, 22.1, 19.3, 15.6, 12.7, 12.1; HRMS ( FAB ⁺ ) : calcd. for C ₆₀ H ₁₀₄ O ₂₇ [M + Na] ⁺ 1279.6663, observed 1279.6659.

< Production Example  9> SPS Synthesis of -2L

<9-1> Synthesis of Compound 2L

Compound 2L was obtained in 90% yield from compound B2 following the general saccharification and deprotection reactions of Examples 1-2 and 1-3. ¹ H NMR (400 MHz, CD ₃ OD): δ 5.36 (d, J = 4.0 Hz, 1H), 4.25 (d, J = 8.0 Hz, 1H), 4.01-3.96 (m, 1H), 3.87-3.84 ( d, J = 12.0 Hz, 1H), 3.70-3.64 (m, 2H), 3.53-3.49 (m, 3H), 3.37-3.12 (m, 6H), 2.01 (d, J = 12.0 Hz, 1H), 1.88 -0.83 (m, 42H), 0.72 (s, 3H); ¹³ C NMR (100 MHz, CD ₃ OD): δ 142.2, 122.8, 104.6. 78.2, 78.0, 75.5, 75.2, 71.7, 68.3, 65.9, 62.9, 58.1, 57.8, 56.1, 51.8 43.6, 41.6, 41.0, 40.8, 37.5, 37.3, 37.1, 37.0, 34.1, 33.5, 31.7, 29.9, 29.5, 29.3 , 27.0, 25.4, 25.1, 23.4, 23.2, 22.1, 19.4, 12.8, 12.5.

<9-2> SPS - 2La's  synthesis

SPS-2La was obtained in a yield of 80% according to the general saccharification reaction of Example 1-2 from compound 2L. ¹ H NMR (400 MHz, CDCl ₃ ): δ 8.24 (d, J = 8.0 Hz, 2H), 8.16-7.78 (m, 24H), 7.72-7.67 (m, 5H), 7.66-7.59 (m, 3H) , 7.55-7.12 (m, 46H), 5.96 (t, J = 8.0 Hz, 1H), 5.91-5.80 (m, 3H), 5.72-5.69 (m, 2H), 5.61-5.42 (m, 8H), 5.01 (d, J = 8.0 Hz, 1H), 4.99-4.92 (m, 2H), 4.87-4.79 (m, 2H), 4.68-4.4.35 (m, 12H), 4.22-4.03 (m, 4H), 3.88 -3.85 (m, 2H), 3.37-3.18 (m, 14H), 2.85 (br s, 1H), 2.75 (br s, 1H), 1.96 (d, J = 12.0 Hz, 1H), 1.90-1.01 (m , 42H), 0.99 (s, 3H), 0.91 (d, J = 8.0 Hz, 3H), 0.88-0.84 (m, 6H), 0.67 (s, 3H); ¹³ C NMR (100 MHz, CDCl ₃ ): δ 166.2, 166.1, 165.9, 165.8, 165.3, 165.2, 165.0, 164.8, 164.5, 141.2, 133.5, 133.3, 133.2, 130.1, 130.0, 129.9, 129.8, 129.7, 129.6, 129.4, 129.0, 128.6, 128.5, 128.4, 128.3, 121.6, 100.1, 100.0, 99.9, 99.8, 98.6, 78.9, 73.4, 73.0, 71.9, 69.7, 65.3, 56.9, 56.3, 50.1, 42.5, 39.9, 39.7, 39.4, 37.4, 37.0, 36.4, 35.9, 32.1, 32.0, 29.9, 28.6, 28.4, 28.2, 24.4, 23.9, 23.0, 22.7, 21.2, 19.5, 18.9, 12.0.

<9-3> SPS Synthesis of -2L

According to the general synthetic procedure for the deprotection reaction of Examples 1-3, SPS-2L was obtained in a yield of 92%. Synthesized. ¹ H NMR (400 MHz, CD ₃ OD): δ 5.37 (d, J = 4.0 Hz, 1H), 4.95 (d, J = 4.0 Hz, 1H), 4.74 (d, J = 8.0 Hz, 1H), 4.67 (d, J = 8.0 Hz, 1H), 4.51 (d, J = 8.0 Hz, 1H), 4.38 (d, J = 8.0 Hz, 1H), 4.29 (d, J = 12.0 Hz, 1H), 4.06 (t , J = 8.0 Hz, 2H), 3.98-3.39 (m, 3H), 3.88-3.83 (m, 6H), 3.71-3.63 (m, 6H), 3.55-3.47 (m, 4H), 3.38-3.18 (m , 28H), 2.10 (d, J = 12.0 Hz, 1H), 1.88-1.76 (m, 4H), 1.70-1.02 (m, 26H), 0.94-0.86 (m, 10H), 0.72 (s, 3H); ¹³ C NMR (100 MHz, CD ₃ OD): δ 142.1, 122.9, 104.9, 103.9, 103.3, 102.5, 82.4, 80.8, 79.9, 78.3, 78.2, 77.9, 76.1, 75.8, 75.3, 75.2, 75.1, 71.8, 71.6 , 71.5, 68.3, 66.1, 63.0, 62.9, 62.7, 58.3, 57.7, 51.8, 43.6, 41.3, 40.8, 40.4, 38.6, 38.1, 37.5, 37.3, 33.4, 33.2, 31.7, 29.7, 29.5, 29.3, 25.47, 25.1 , 23.3, 23.1, 22.3, 20.1, 19.4, 12.5; HRMS ( FAB ⁺ ) : calcd. for C ₆₀ H ₁₀₂ O ₂₇ [M + Na] ⁺ 1277.6506, observed 1277.6503.

< Production Example  10> SPS Synthesis of -3L

<10-1> Synthesis of Compound 3L

Compound 3L was obtained in 87% yield from compound B3 following the general saccharification and deprotection reactions of Examples 1-2 and 1-3. ¹ H NMR (400 MHz, CD ₃ OD): δ 5.36 (d, J = 4.0 Hz, 1H), 4.25 (d, J = 8.0 Hz, 1H), 4.01-3.96 (m, 1H), 3.87-3.84 ( d, J = 12.0 Hz, 1H), 3.70-3.64 (m, 2H), 3.53-3.49 (m, 3H), 3.37-3.12 (m, 6H), 2.01 (d, J = 12.0 Hz, 1H), 1.20 -0.86 (m, 47H), 0.72 (s, 3H); ¹³ C NMR (100 MHz, CD ₃ OD): δ 142.2, 122.8, 104.6. 78.2, 78.0, 75.5, 75.2, 71.7, 68.3, 65.9, 62.9, 58.1, 57.8, 56.1, 51.8 43.6, 41.6, 41.0, 40.8, 38.6, 37.5, 37.3, 37.1, 37.0, 35.2, 34.1, 33.5, 31.7, 30.5 , 29.9, 29.5, 29.3, 27.0, 25.4, 25.1, 24.3, 23.4, 23.2, 22.1, 20.3, 20.1, 19.6, 19.4, 12.5.

<10-2> SPS - Of 3La  synthesis

SPS-3La was obtained in a yield of 78% according to the general saccharification reaction of Example 1-2 from compound 3L. ¹ H NMR (400 MHz, CDCl ₃ ): δ 8.24 (d, J = 8.0 Hz, 2H), 8.15-7.78 (m, 24H), 7.72-7.67 (m, 5H), 7.66-7.59 (m, 3H) , 7.59-7.12 (m, 46H), 5.96 (t, J = 8.0 Hz, 1H), 5.92-5.82 (m, 3H), 5.72-5.69 (m, 2H), 5.61-5.42 (m, 8H), 5.01 (d, J = 8.0 Hz, 1H), 4.98-4.92 (m, 2H), 4.87-4.79 (m, 2H), 4.68-4.4.35 (m, 12H), 4.22-4.03 (m, 4H), 3.88 -3.85 (m, 2H), 3.37-3.18 (m, 14H), 2.85 (br s, 1H), 2.75 (br s, 1H), 1.96 (d, J = 12.0 Hz, 1H), 1.90-1.01 (m , 47H), 0.99 (s, 3H), 0.91 (d, J = 8.0 Hz, 3H), 0.88-0.84 (m, 6H), 0.68 (s, 3H); ¹³ C NMR (100 MHz, CDCl ₃ ): δ 166.2, 166.1, 160.0, 165.9, 165.8, 165.3, 165.2, 165.0, 164.8, 164.5, 141.2, 133.5, 133.3, 133.2, 130.1, 130.0, 129.9, 129.8, 129.7, 129.6, 129.4, 129.0, 128.6, 128.5, 128.4, 128.3, 121.6, 100.1, 100.0, 99.9, 99.8, 98.6, 78.9, 73.4, 73.0, 72.0, 69.7, 65.3, 56.9, 56.3, 50.1, 42.5, 39.9, 39.7, 39.4, 37.4, 37.0, 36.4, 35.9, 32.1, 32.0, 29.9, 28.6, 28.4, 28.2, 24.4, 23.9, 23.0, 22.7, 21.2, 19.5, 18.9, 12.0, 11.5.

<10-3> SPS Synthesis of -3L

According to the general synthetic procedure for the deprotection reaction of Examples 1-3, SPS-3L was obtained in a yield of 91%. Synthesized. ¹ H NMR (400 MHz, CD ₃ OD): δ 5.38 (d, J = 4.0 Hz, 1H), 4.96 (d, J = 4.0 Hz, 1H), 4.75 (d, J = 8.0 Hz, 1H), 4.68 (d, J = 8.0 Hz, 1H), 4.52 (d, J = 8.0 Hz, 1H), 4.40 (d, J = 8.0 Hz, 1H), 4.30 (d, J = 12.0 Hz, 1H), 4.10 (t , J = 8.0 Hz, 2H), 3.98-3.39 (m, 3H), 3.88-3.83 (m, 6H), 3.71-3.63 (m, 6H), 3.55-3.47 (m, 4H), 3.38-3.18 (m , 28H), 2.38 (d, J = 12.0 Hz, 1H), 2.10 (d, J = 12.0 Hz, 1H), 1.88-1.76 (m, 4H), 1.70-1.02 (m, 28H), 0.94-0.86 ( m, 10H), 0.72 (s, 3H); ¹³ C NMR (100 MHz, CD ₃ OD): δ 142.1, 122.9, 104.9, 103.9, 103.3, 102.5, 82.4, 80.8, 79.9, 78.3, 78.2, 77.9, 76.1, 75.8, 75.3, 75.2, 75.1, 71.8, 71.6 , 71.5, 68.3, 66.1, 63.0, 62.9, 62.7, 58.3, 57.7, 51.8, 43.6, 41.3, 40.8, 40.4, 38.6, 38.1, 37.5, 37.3, 35.3, 33.4, 33.2, 30.5, 29.7, 29.5, 27.3, 25.5 , 24.3, 22.3, 20.3, 20.1, 19.6, 19.5, 12.5, 12.4; HRMS (FAB ⁺ ) : calcd. for C ₆₂ H ₁₀₆ O ₂₇ [M + Na] ⁺ 1305.6819, observed 1305.6824.

< Production Example  11> SPS Synthesis of -4L

<11-1> Synthesis of Compound 4L

Compound 4L was obtained in compound yield of 88% according to the general saccharification and deprotection reactions of Examples 1-2 and 1-3. ¹ H NMR (400 MHz, CD ₃ OD): δ 5.37 (d, J = 4.0 Hz, 1H), 4.42-4.36 (m, 1H), 4.25 (d, J = 8.0 Hz, 1H), 4.01-3.96 ( m, 1H), 3.87-3.84 (d, J = 12.0 Hz, 1H), 3.70-3.64 (m, 2H), 3.53-3.49 (m, 3H), 3.45-3.43 (m, 6H), 3.37-3.12 ( m, 6H), 2.39 (d, J = 12.0 Hz, 1H), 2.18-0.95 (m, 37H), 0.76 (s, 6H); ¹³ C NMR (100 MHz, CD ₃ OD): δ 142.2, 122.6, 110.7, 104.6, 82.3, 80.6, 78.2, 78.0, 75.2, 71.8, 68.1, 67.9, 66.2, 63.9, 62.9, 57.9, 55.0, 51.8, 43.0 , 41.0, 40.3, 38.5, 38.2, 33.3, 32.9, 32.8, 32.5, 31.6, 30.0, 29.6, 24.4, 22.1, 20.0, 17.6, 16.9, 15.1.

<11-2> SPS - 4La's  synthesis

From the compound 4L, SPS-4La was obtained in a yield of 78% according to the general saccharification reaction of Example 1-2. ¹ H NMR (400 MHz, CDCl ₃ ): δ 8.25 (d, J = 8.0 Hz, 2H), 8.15-7.77 (m, 24H), 7.72-7.67 (m, 5H), 7.66-7.59 (m, 3H) , 7.58-7.12 (m, 46H), 5.96 (t, J = 8.0 Hz, 1H), 5.92-5.82 (m, 3H), 5.71-5.69 (m, 3H), 5.61-5.42 (m, 8H), 5.01 (d, J = 8.0 Hz, 1H), 4.98-4.92 (m, 2H), 4.88-4.79 (m, 2H), 4.69-4.4.35 (m, 12H), 4.22-4.03 (m, 4H), 3.87 -3.85 (m, 2H), 3.37-3.18 (m, 14H), 2.85 (br s, 1H), 2.75 (br s, 1H), 1.96 (d, J = 12.0 Hz, 1H), 1.90-1.01 (m , 47H), 0.99 (s, 3H), 0.91 (d, J = 8.0 Hz, 3H), 0.88-0.84 (m, 6H), 0.70 (s, 6H); ¹³ C NMR (100 MHz, CDCl ₃ ): δ 166.2, 166.1, 166.0, 165.9, 165.8, 165.2, 165.1, 164.6, 141.2, 133.5, 133.2, 133.1, 133.0, 130.3, 130.0, 129.9, 128.8, 129.8, 129.7, 129.6, 129.3, 129.2, 129.0, 128.9, 128.6, 128.5, 128.4, 128.3, 121.2, 109.4, 109.3, 101.2, 101.1, 100.0, 99.9, 99.8, 80.9, 79.0, 72.9, 72.4, 72.3, 72.2, 67.0, 66.4, 62.2, 56.6, 53.6, 50.1, 41.7, 40.4, 39.9, 39.4, 37.2, 37.1, 32.2, 31.9, 31.7, 31.5, 30.5, 30.4, 28.6, 20.9, 19.5, 17.3, 16.4, 14.7.

<11-3> SPS Synthesis of -4L

Following the general synthetic procedure for the deprotection reaction of Examples 1-3, SPS-4L was obtained at a yield of 91%. Synthesized. ¹ H NMR (400 MHz, CD ₃ OD): δ 5.36 (d, J = 4.0 Hz, 1H), 4.92 (d, J = 4.0 Hz, 1H), 4.72 (d, J = 8.0 Hz, 1H), 4.64 (d, J = 8.0 Hz, 1H), 4.48 (d, J = 8.0 Hz, 1H), 4.36 (d, J = 8.0 Hz, 1H), 4.26 (d, J = 12.0 Hz, 1H), 4.04 (t , J = 8.0 Hz, 2H), 3.95-3.36 (m, 3H), 3.88-3.83 (m, 6H), 3.71-3.63 (m, 6H), 3.55-3.47 (m, 4H), 3.38-3.18 (m , 28H), 2.36 (d, J = 12.0 Hz, 1H), 2.13 (d, J = 12.0 Hz, 1H), 2.01-1.80 (m, 7H), 1.74-1.35 (m, 12H), 1.29-1.12 ( m, 5H), 1.0 (s, 3H), 0.93 (d, J = 8.0 Hz, 4H), 0.78 (s, 6H); ¹³ C NMR (100 MHz, CD ₃ OD): δ 142.2, 122.7, 110.7, 105.1, 104.9, 103.9, 103.3, 102.5, 82.3, 80.7, 79.9, 78.2, 78.1, 77.9, 75.8, 75.3, 75.2, 75.0, 71.8 , 71.6, 71.4, 69.7, 68.3, 68.0, 66.1, 63.9, 63.1, 62.7, 57.9, 51.7, 43.0, 41.5, 41.1, 40.4, 38.5, 38.2, 33.3, 33.0, 32.9, 32.6, 31.7, 31.6, 30.0, 29.7 , 22.1, 20.1, 17.6, 16.9, 15.7, 15.1; HRMS ( FAB ⁺ ) : calcd. for C ₆₀ H ₉₈ O ₂₉ [M + Na] ⁺ 1305.6091, observed 1305.6089.

< Example  3> SMAs  Synthesis method of

The synthesis scheme of SMAs is shown in FIG. 3. Compounds of three SMA-As, two SMA-Es, and three SMA-Ds were synthesized according to the synthesis methods of the following <3-1> to <3-3>.

<3-1-1> Synthesis of Compound A (Steps i and ii of FIG. 3)

A mixture of tosylate cholesterol (2 mmol) and 2-propen-1-ol / 3-butene-1-ol or 4-penten-1-ol (40 mmol) mixed in toluene (2 mL) was then 80-90 Stir at nitrogen for 4 hours under nitrogen. After cooling to room temperature, CH ₂ Cl ₂ (30 mL) was added to the mixture. The mixture was washed with brine (30 mL). After separation, an additional 20 mL of CH ₂ Cl ₂ was added to the organic layer. The organic layer was then washed with brine (30 mL), dried and evaporated to dryness. The crude product was stirred vigorously in a 5: 3: 1 acetone / water / tert-butyl alcohol mixture at room temperature under ambient atmosphere. Here 50 wt.% Of N-methylmorpholine N-oxide (NMO) mixed in water (1.10 mmol) was added. OsO ₄ (0.25 mol-%) was added after the% solution was added. When the starting material disappeared according to TLC analysis, the solvent was removed under vacuum at 50 ° C. The residue was evaporated twice with toluene at 40 ° C. under vacuum to afford the crude product which was purified by flash column chromatography eluting with ethyl acetate-hexanes to afford the desired compound A.

<3-1-2> Synthesis of Compound B (Steps iii, iv of Figure 3)

2,2-dimethyl-1,3-dioxolane-4-methanol / 2,2-dimethyl-1,3-dioxolane-4-ethanol or 3- (2,2-dimethyl-1,3-dioxolane- To a solution of 4-yl) propan-1-ol (12.9 mmol) NaH (60% in mineral oil, 26 mmol) was added and the mixture was stirred at room temperature under inert atmosphere for 30 minutes. Cholesteryl tosylate (16 mmol) or diosgenyl tosylate (16 mmol) in THF (10 mL) was then added dropwise to the reaction mixture. After addition, the mixture was refluxed under nitrogen overnight and cooled to room temperature. Excess NaH was removed by carefully adding water to the mixture. The reaction mixture was diluted with CH ₂ Cl ₂ (50 mL), washed with water and brine, dried over sodium sulfate, filtered and concentrated under reduced pressure. The intermediate residue was dissolved in 20 ml of THF-aqueous solution (15: 5) with p-toluenesulfonic acid (160 mg) and the resulting solution was refluxed overnight. After extraction twice with CH ₂ Cl ₂ , it was washed once with water, once with brine solution, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by flash column chromatography eluting with ethyl acetate-hexanes to afford the desired compound B.

<3-2> Of maltosylation reactions  General Synthesis Procedure (Step v of Figure 3)

A mixture of the compound, 2,4,5-collidine (1.0 equiv) and AgOTf (2.4 equiv), produced in Example 3-1, was stirred at -45 ° C in anhydrous CH ₂ Cl ₂ . A solution of perbenzoylated maltosylbromide / perbenzoylated glucosylbromide in anhydrous CH ₂ Cl ₂ was added dropwise to this suspension for 30 minutes. After stirring was continued at −45 ° C. for 30 minutes, the temperature of the reaction mixture was increased to 0 ° C. and stirring was continued for 1 hour. After completion of the reaction (indicated by TLC), the reaction was quenched by addition of pyridine, then diluted with CH ₂ Cl ₂ and filtered over celite. The filtrate was washed successively with 1 M aqueous Na ₂ S ₂ O ₃ solution (40 mL), 0.1 M aqueous HCl solution (40 mL) and brine (2 × 40 mL). The organic layer was dried over anhydrous Na ₂ SO _4, and then the solvent was removed with a rotary evaporator. The residue obtained was purified by silica gel column chromatography (EtOAc / hexanes) to afford the desired compound as a white solid.

<3-3> Deprotection Vaporization  reaction ( deprotection  General synthetic procedure for the reaction (step vi of FIG. 3)

This was followed by the synthetic method of Chae, PS et al. (Nat Meth 2010, 7, 1003.). De-O-benzoylation was performed under Zemplen's conditions. The O-protected compound was dissolved in anhydrous CH ₂ Cl ₂ and then MeOH was added slowly until a continuous precipitation appeared. To the reaction mixture was added 0.5 M of methanolic solution, NaOMe, to a final concentration of 0.05 M. The reaction mixture was stirred at room temperature for 6 hours. After completion of the reaction, the reaction mixture was neutralized with Amberlite IR-120 (H + form) resin. The resin was removed by filtration, washed with MeOH and the solvent was removed from the filtrate in vacuo. The residue was purified by silica gel chromatography (MeOH: CH ₂ Cl ₂ ) to afford the title compound as a white solid.

< Production Example  12> SMA Synthesis of -A1

<12-1> Synthesis of Compound A1

Compound A1 was synthesized in an 85% yield according to the synthesis procedure of Example 3-1-1. ¹ H NMR (400 MHz, CDCl ₃ ): δ 3.86-3.82 (m, 1H), 3.73-3.62 (m, 2H), 3.55 (br s, 1H), 3.50-3.41 (m, 2H), 3.05 (br s, 2H), 1.97-1.94 (m, 1H), 1.83-1.74 (m, 2H), 1.62-0.97 (m, 28H), 0.90-0.85 (m, 10H), 0.77 (s, 3H), 0.64 ( s, 3H)); ¹³ C NMR (100 MHz, CDCl ₃ ): δ 75.1, 70.8, 69.8, 64.6, 56.6, 56.4, 54.4, 42.7, 40.2, 39.8, 39.7, 36.3, 36.0, 35.9, 36.6, 33.1, 32.9, 32.1, 28.7, 28.4, 28.2, 25.8, 24.3, 24.0, 22.9, 22.7, 20.9, 18.8, 12.2, 11.5.

<12-2> SMA - Of A1a  synthesis

Compound SMA-A1a was synthesized in a yield of 85% according to the general saccharification reaction procedure of Example 3-2. ¹ H NMR (400 MHz, CDCl ₃ ): δ 8.21-7.98 (m, 10H), 7.91-7.70 (m, 18H), 7.54-7.12 (m, 42H), 6.23-6.15 (m, 2H), 5.80- 5.69 (m, 6H), 5.35-5.22 (m, 4H), 4.82-4.66 (m, 4H), 4.57-4.30 (m, 10H), 3.95-3.82 (m, 2H), 3.72-3.68 (m, 2H ), 3.23 (br s, 1H), 3.01-2.95 (m, 1H), 2.85-2.77 (m, 1H), 1.97-1.94 (m, 1H), 1.83-1.74 (m, 2H), 1.62-0.97 ( m, 28H), 0.92-0.85 (m, 10H), 0.76 (s, 3H), 0.64 (s, 3H)); ¹³ C NMR (100 MHz, CDCl ₃ ): δ 166.2, 166.1, 166.0, 165.9, 165.8, 165.7, 165.6, 165.5, 165.3, 165.2, 165.0, 133.5, 133.3, 133.2, 130.1, 129.9, 129.8, 129.7, 129.6, 129.5, 129.4, 129.0, 128.9, 128.5, 128.7, 128.6, 128.5, 128.4, 128.3, 128.2, 128.1, 101.0, 96.4, 74.6, 74.5, 73.4, 73.1, 72.8, 72.6, 71.0, 69.3, 63.6, 62.6, 60.5, 56.5, 56.3, 54.0, 42.7, 40.1, 39.6, 36.3, 35.9, 35.8, 35.7, 35.6, 32.0, 28.4, 28.1, 24.3, 24.0, 22.9, 22.7, 20.8, 18.8, 14.3, 12.2, 11.4, 11.3.

<12-3> SMA Synthesis of -A1

Following the general synthetic procedure for the deprotection reaction of Example 3-3, SMA-A1 was obtained in a yield of 92%. Synthesized. ¹ H NMR (400 MHz, CD ₃ OD): δ 5.18 (d, J = 4.0 Hz, 2H), 4.66 and 4.60 (d, J = 8.0 Hz, 1H), 4.45 and 4.40 (d, J = 8.0 Hz, 1H), 4.12-4.05 (m, 2H), 3.93-3.80 (m, 8H), 3.72-3.51 (m, 10H), 3.47-3.39 (m, 5H), 3.35-3.25 (m, 4H), 2.03 ( d, J = 12.0 Hz, 1H), 1.87-1.80 (m, 2H), 1.61-1.01 (m, 28H), 0.96-0.89 (m, 10H), 0.84 (s, 3H), 0.70 (s, 3H) ; ¹³ C NMR (100 MHz, CD ₃ OD): δ 105.0, 104.6, 104.5, 104.0, 103.0, 98.3, 93.9, 81.6, 81.5, 81.3, 79.4, 78.7, 77.8, 76.8, 76.7, 76.3, 76.1, 75.2, 75.1 , 75.0, 74.9, 74.5, 74.3, 71.6, 69.1, 62.9, 62.4, 58.1, 58.0, 57.8, 56.0, 55.9, 43.9, 41.6, 40.9, 40.8, 37.5, 37.3, 37.1, 37.0, 34.1, 34.0, 33.5, 29.8 , 29.5, 29.3, 26.9, 26.7, 25.4, 25.1, 19.8, 12.8, 11.9; HRMS ( FAB ⁺ ) : calcd. for C ₅₄ H ₉₄ O ₂₃ [M + Na] ⁺ 1133.6084, observed 1133.6079.

< Production Example  13> SMA Synthesis of -A2

<13-1> Synthesis of Compound A2

Compound A2 was synthesized in an 85% yield according to the synthesis procedure of Example 3-1-1. ¹ H NMR (400 MHz, CDCl ₃ ): δ 3.93-3.88 (m, 1H), 3.64-3.51 (m, 5H), 2.86 (br s, 2H), 1.96-1.92 (m, 1H), 1.84-0.99 (m, 32H), 0.90-0.85 (m, 10H), 0.76 (s, 3H), 0.64 (s, 3H)); ¹³ C NMR (100 MHz, CDCl ₃ ): δ 75.1, 72.4, 72.2, 66.7, 66.6, 66.4, 56.6, 56.4, 54.3, 42.7, 40.1, 39.9, 39.8, 39.7, 36.4, 36.0, 35.9, 35.6, 33.1, 33.0, 32.9, 32.1, 28.8, 28.4, 28.9, 28.2, 25.8, 24.3, 24.0, 22.9, 22.7, 20.9, 18.8, 12.2, 11.5.

<13-2> SMA - Of A2a  synthesis

Compound SMA-A2a was synthesized in a yield of 85% according to the general saccharification reaction procedure of Example 3-2. ¹ H NMR (400 MHz, CDCl ₃ ): δ 8.20-7.98 (m, 10H), 7.91-7.70 (m, 18H), 7.54-7.12 (m, 42H), 6.23-6.15 (m, 2H), 5.80- 5.69 (m, 6H), 5.35-5.22 (m, 4H), 4.82-4.66 (m, 4H), 4.57-4.30 (m, 10H), 3.95-3.82 (m, 2H), 3.72-3.67 (m, 2H ), 3.23 (br s, 1H), 3.01-2.94 (m, 1H), 2.85-2.77 (m, 1H), 1.96-1.92 (m, 1H), 1.84-0.99 (m, 32H), 0.91-0.85 ( m, 10H), 0.76 (s, 3H), 0.65 (s, 3H)); ¹³ C NMR (100 MHz, CDCl ₃ ): δ 166.2, 166.1, 166.0, 165.9, 165.8, 165.7, 165.6, 165.5, 165.3, 165.2, 165.0, 133.5, 133.3, 133.2, 130.1, 129.9, 129.8, 129.7, 129.6, 129.5, 129.4, 129.0, 128.9, 128.5, 128.7, 128.6, 128.5, 128.4, 128.3, 128.2, 128.1, 101.0, 96.4, 70.1, 69.2, 63.6, 62.6, 60.4, 56.5, 56.3, 54.1, 42.6, 39.6, 39.5, 36.3, 35.9, 35.8, 35.5, 32.7, 32.0, 28.3, 28.1, 24.3, 23.9, 22.9, 22.7, 21.1, 18.8, 14.3, 12.2, 11.4, 11.3.

<13-3> SMA Synthesis of -A2

SMA-A2 in 92% yield according to the general synthetic procedure for deprotection reaction of Example 3-3. Synthesized. ¹ H NMR (400 MHz, CD ₃ OD): δ 5.18 (d, J = 4.0 Hz, 2H), 4.65 and 4.51 (d, J = 8.0 Hz, 1H), 4.40 (d, J = 8.0 Hz, 1H) , 4.10-4.07 (m, 2H), 3.92-3.78 (m, 8H), 3.70-3.51 (m, 10H), 3.47-3.37 (m, 5H), 3.32-3.25 (m, 4H), 2.03 (d, J = 12.0 Hz, 1H), 1.90-1.78 (m, 2H), 1.70-1.01 (m, 30H), 0.96-0.89 (m, 10H), 0.84 (s, 3H), 0.70 (s, 3H); ¹³ C NMR (100 MHz, CD ₃ OD): δ 104.8, 104.7, 104.5, 104.2, 103.0, 98.3, 93.9, 81.9, 81.5, 81.4, 81.3, 78.1, 77.9, 76.7, 76.6, 75.5, 75.2, 74.8, 74.4 , 74.2, 73.6, 73.1, 71.7, 71.6, 64.5, 65.2, 62.8, 62.3, 58.1, 57.8, 56.0, 43.9, 41.6, 41.1, 40.9, 40.8, 37.5, 37.3, 37.1, 37.0, 34.2, 33.9, 33.6, 33.5 , 29.9, 29.5, 29.3, 27.2, 26.9, 25.4, 25.2, 23.4, 23.1, 22.0, 19.4, 12.8, 12.1; HRMS ( FAB ⁺ ) : calcd. for C ₅₅ H ₉₆ O ₂₃ [M + Na] ⁺ 1147.6240, observed 1147.6235.

< Production Example  14> SMA Synthesis of -A3

<14-1> Synthesis of Compound A3

Compound A3 was synthesized in an 85% yield according to the synthesis procedure of Example 3-1-1. ¹ H NMR (400 MHz, CDCl ₃ ): δ 3.70-3.65 (m, 1H), 3.60-3.41 (m, 4H), 3.25-3.20 (m, 1H), 1.97-1.92 (m, 1H), 1.84- 0.99 (m, 34H), 0.90-0.85 (m, 10H), 0.76 (s, 3H), 0.64 (s, 3H)); ¹³ C NMR (100 MHz, CDCl ₃ ): δ 79.0, 72.1, 68.2, 66.8, 56.6, 56.5, 54.5, 44.9, 42.7, 40.2, 39.6, 37.1, 36.3, 35.9, 35.6, 34.9, 34.8, 32.2, 31.1, 28.9, 28.4, 28.3, 28.1, 26.8, 26.7, 24.3, 24.0, 22.9, 22.7, 21.4, 18.8, 12.4, 12.2.

<14-2> SMA - Of A3a  synthesis

Compound SMA-A3a was synthesized in a yield of 85% according to the general saccharification reaction procedure of Example 3-2. ¹ H NMR (400 MHz, CDCl ₃ ): δ 8.21-7.98 (m, 10H), 7.91-7.70 (m, 18H), 7.54-7.12 (m, 42H), 6.23-6.15 (m, 2H), 5.80- 5.69 (m, 6H), 5.35-5.22 (m, 4H), 4.82-4.66 (m, 4H), 4.57-4.30 (m, 10H), 3.95-3.82 (m, 2H), 3.72-3.65 (m, 2H ), 3.23 (br s, 1H), 3.01-2.94 (m, 1H), 2.85-2.77 (m, 1H), 1.97-1.92 (m, 1H), 1.84-0.99 (m, 34H), 0.90-0.84 ( m, 10H), 0.76 (s, 3H), 0.66 (s, 3H)); ¹³ C NMR (100 MHz, CDCl ₃ ): δ 166.2, 166.1, 166.0, 165.9, 165.8, 165.7, 165.6, 165.5, 165.3, 165.2, 165.0, 133.5, 133.3, 133.2, 130.1, 129.9, 129.8, 129.7, 129.6, 129.5, 129.4, 129.0, 128.9, 128.8, 128.7, 128.6, 128.5, 128.4, 128.3, 128.2, 128.1, 101.0, 96.4, 78.2, 71.4, 70.0, 69.2, 69.1, 60.3, 56.5, 56.3, 54.4, 44.8, 42.6, 40.1, 39.5, 36.9, 36.2, 35.8, 35.7, 35.5, 34.9, 32.2, 28.9, 28.3, 28.2, 28.0, 24.2, 23.8, 22.9, 22.6, 21.3, 21.0, 18.8, 14.2, 12.3, 12.2, 11.5.

<14-3> SMA Synthesis of -A3

SMA-A3 at 94% yield according to the general synthetic procedure for deprotection reaction of Example 3-3 Synthesized. ¹ H NMR (400 MHz, CD ₃ OD): δ 5.17 (d, J = 4.0 Hz, 2H), 4.65 and 4.51 (d, J = 8.0 Hz, 1H), 4.39 (d, J = 8.0 Hz, 1H) , 4.10-4.07 (m, 2H), 3.94-3.78 (m, 8H), 3.72-3.51 (m, 10H), 3.48-3.36 (m, 5H), 3.32-3.25 (m, 4H), 2.03 (d, J = 12.0 Hz, 1H), 1.90-1.78 (m, 2H), 1.72-1.01 (m, 32H), 0.96-0.89 (m, 10H), 0.84 (s, 3H), 0.70 (s, 3H); ¹³ C NMR (100 MHz, CD ₃ OD): δ 104.8, 104.7, 104.6, 104.2, 103.0, 98.3, 93.9, 81.9, 81.5, 81.4, 81.3, 78.1, 77.9, 76.7, 76.6, 75.5, 75.2, 74.8, 74.4 , 74.2, 73.6, 73.1, 71.7, 71.6, 64.6, 65.2, 62.8, 62.3, 58.1, 57.8, 56.0, 43.9, 41.6, 41.1, 40.9, 40.8, 37.5, 37.3, 37.1, 37.0, 34.2, 33.9, 33.6, 33.5 , 29.9, 29.5, 29.3, 27.2, 26.9, 25.5, 25.2, 23.4, 23.1, 22.0, 19.4, 12.7, 12.1; HRMS ( FAB ⁺ ) : calcd. for C ₅₆ H ₉₈ O ₂₃ [M + Na] ⁺ 1161.6397, observed 1161.6393.

< Production Example  15> SMA Synthesis of -E1

<15-1> Synthesis of Compound B1

Compound B1 was synthesized in an 80% yield according to the synthesis procedure of Example 3-1-2. ¹ H NMR (400 MHz, CDCl ₃ ): δ 5.32 (d, J = 4.0 Hz, 1H), 3.90-3.50 (m, 5H), 3.20-3.12 (m, 1H), 2.50-2.46 (m, 1H) , 2.01-1.85 (m, 5H), 1.62-1.29 (m, 12H), 1.20-1.09 (m, 8H), 1.03-0.88 (m, 16H), 0.68 (s, 3H); ¹³ C NMR (100 MHz, CDCl ₃ ): δ 140.1, 121.5, 79.8, 72.2, 71.8, 70.8, 69.8, 64.6, 56.6, 56.4, 54.4, 42.7, 40.2, 39.8, 39.7, 36.3, 36.0, 35.9, 36.6, 33.1, 32.9, 32.1, 28.7, 28.4, 28.2, 25.8, 24.3, 24.0, 22.9, 22.7, 20.9, 18.8, 12.2.

<15-2> SMA - Of E1a  synthesis

Compound SMA-E1a was synthesized in 84% yield following the general saccharification reaction procedure of Example 3-2. ¹ H NMR (400 MHz, CDCl ₃ ): δ 8.18-7.98 (m, 10H), 7.90-7.70 (m, 18H), 7.54-7.12 (m, 42H), 6.23-6.15 (m, 2H), 5.80- 5.69 (m, 6H), 5.35-5.22 (m, 4H), 4.82-4.66 (m, 5H), 4.57-4.30 (m, 10H), 3.95-3.82 (m, 2H), 3.72-3.68 (m, 2H ), 3.23 (br s, 1H), 3.01-2.95 (m, 1H), 2.50-2.46 (m, 1H), 2.01-1.85 (m, 5H), 1.62-1.29 (m, 12H), 1.20-1.09 ( m, 8H), 1.03-0.88 (m, 16H), 0.68 (s, 3H); ¹³ C NMR (100 MHz, CDCl ₃ ): δ 166.2, 166.1, 166.0, 165.9, 165.8, 165.7, 165.6, 165.5, 165.2, 165.1, 165.0, 164.9, 140.6, 133.4, 133.1, 129.9, 129.7, 128.4, 128.3, 121.5, 100.5, 96.6, 79.4, 73.3, 73.1, 72.8, 71.3, 70.0, 69.8, 69.2, 69.1, 68.2, 63.5, 62.6, 60.4, 56.8, 56.2, 50.2, 50.1, 42.4, 39.8, 39.6, 36.7, 36.6, 36.3, 35.8, 31.9, 31.8, 28.3, 28.0, 24.3, 23.8, 22.9, 22.6, 21.0, 18.8, 12.2.

<14-3> SMA Synthesis of -E1

Following the general synthetic procedure for the deprotection reaction of Example 3-3, SMA-E1 was obtained in a yield of 93%. Synthesized. ¹ H NMR (400 MHz, CD ₃ OD): δ 5.37 (d, J = 4.0 Hz, 1H), 5.17 (d, J = 4.0 Hz, 2H), 4.53-4.48 (m, 1H), 4.42 and 4.38 ( d, J = 8.0 Hz, 1H), 4.08-3.99 (m, 2H), 3.92-3.75 (m, 8H), 3.70-3.60 (m, 10H), 3.58-3.35 (m, 5H), 3.30-3.24 ( m, 4H), 2.42-2.36 (m, 1H), 2.20-1.85 (m, 5H), 1.61-1.29 (m, 12H), 1.20-1.10 (m, 8H), 1.03-0.88 (m, 16H), 0.73 (s, 3 H); ¹³ C NMR (100 MHz, CD ₃ OD): δ 142.1, 122.9, 104.9, 104.6, 104.3, 103.0, 81.3, 77.9, 77.8, 76.8, 76.7, 75.2, 74.9, 74.8, 74.3, 71.6, 62.9, 62.4, 58.3 , 57.7, 51.8, 43.6, 41.3, 40.8, 40.2, 38.5, 38.1, 37.5, 37.3, 33.4, 33.2, 29.5, 29.4, 29.3, 25.5, 25.1, 23.3, 23.1, 22.3, 20.0, 19.4, 12.5; HRMS ( FAB ⁺ ) : calcd. for C ₅₄ H ₉₂ O ₂₃ [M + Na] ⁺ 1131.5927, observed 1131.5930.

< Production Example  16> SMA Synthesis of -E3

<16-1> Synthesis of Compound B2

Compound B2 was synthesized in an 80% yield according to the synthesis procedure of Example 3-1-2. ¹ H NMR (400 MHz, CDCl ₃ ): δ 5.33 (d, J = 4.0 Hz, 1H), 3.90-3.50 (m, 5H), 3.20-3.12 (m, 1H), 2.48-2.45 (m, 1H) , 2.02-1.85 (m, 5H), 1.62-1.30 (m, 16H), 1.20-1.09 (m, 8H), 1.03-0.88 (m, 16H), 0.67 (s, 3H); ¹³ C NMR (100 MHz, CDCl ₃ ): δ 140.6, 121.8, 79.4, 72.1, 68.2, 66.8, 56.6, 56.5, 54.5, 44.9, 42.7, 40.2, 39.6, 37.1, 36.3, 35.9, 35.6, 34.9, 34.8, 32.2, 31.1, 28.9, 28.4, 28.3, 28.1, 26.8, 26.7, 24.3, 24.0, 22.9, 22.5, 21.4, 18.9, 12.1.

<16-2> SMA - Of E3a  synthesis

Compound SMA-E3a was synthesized in 84% yield following the general saccharification reaction procedure of Example 3-2. ¹ H NMR (400 MHz, CDCl ₃ ): δ 8.16-7.98 (m, 10H), 7.90-7.70 (m, 18H), 7.54-7.12 (m, 42H), 6.23-6.15 (m, 2H), 5.80- 5.69 (m, 6H), 5.35-5.22 (m, 4H), 4.82-4.66 (m, 5H), 4.57-4.30 (m, 10H), 3.95-3.82 (m, 2H), 3.72-3.68 (m, 2H ), 3.23 (br s, 1H), 3.01-2.95 (m, 1H), 2.50-2.46 (m, 1H), 2.01-1.85 (m, 5H), 1.62-1.29 (m, 14H), 1.20-1.09 ( m, 10H), 1.03-0.88 (m, 16H), 0.67 (s, 3H); ¹³ C NMR (100 MHz, CDCl ₃ ): δ 166.3, 166.1, 166.0, 165.9, 165.8, 165.7, 165.6, 165.5, 165.2, 165.1, 165.0, 164.9, 140.6, 133.4, 133.1, 129.9, 129.7, 128.4, 128.3, 121.5, 100.5, 96.6, 79.4, 73.3, 73.1, 72.8, 71.3, 70.0, 69.8, 69.2, 69.1, 68.2, 63.5, 62.6, 60.4, 56.8, 56.2, 50.2, 50.1, 42.4, 39.8, 39.6, 36.8, 36.7, 36.6, 36.3, 35.8, 31.9, 31.8, 28.3, 28.0, 24.3, 23.8, 22.9, 22.6, 21.0, 18.8, 14.2, 12.2.

<16-3> SMA Synthesis of -E3

SMA-E3 at 94% yield according to the general synthetic procedure for deprotection reaction of Example 3-3 Synthesized. ¹ H NMR (400 MHz, CD ₃ OD): δ 5.37 (d, J = 4.0 Hz, 1H), 5.17 (d, J = 4.0 Hz, 2H), 4.53-4.48 (m, 1H), 4.42 and 4.38 ( d, J = 8.0 Hz, 1H), 4.10-3.99 (m, 2H), 3.92-3.74 (m, 8H), 3.70-3.60 (m, 10H), 3.58-3.35 (m, 5H), 3.30-3.24 ( m, 4H), 2.42-2.35 (m, 1H), 2.20-1.85 (m, 5H), 1.61-1.25 (m, 16H), 1.20-1.10 (m, 8H), 1.03-0.88 (m, 16H), 0.73 (s, 3 H); ¹³ C NMR (100 MHz, CD ₃ OD): δ 142.1, 122.8, 103.1, 98.3, 81.6, 81.4, 77.9, 76.8, 75.3, 75.2, 74.9, 74.3, 71.6, 67.0, 62.8, 62.5, 58.3, 57.7, 51.8 , 43.6, 41.3, 40.8, 40.4, 38.6, 38.1, 37.5, 37.2, 33.4, 33.2, 29.6, 29.5, 29.3, 25.4, 25.1, 22.3, 20.0, 19.4, 15.6, 12.5; HRMS ( FAB ⁺ ) : calcd. for C ₅₆ H ₉₆ O ₂₃ [M + Na] ⁺ 1159.6240, observed 1159.6234.

< Production Example  17> SMA Synthesis of -D1

<17-1> Synthesis of Compound C1

Compound C1 was synthesized in an 80% yield according to the synthesis procedure of Example 3-1-2. ¹ H NMR (400 MHz, CDCl ₃ ): δ 5.34 (d, J = 4.0 Hz, 1H), 4.42 (q, J = 7.6 Hz, 1H), 3.84 (br s, 1H), 3.72-3.68 (m, 1H), 3.62-3.45 (m, 4H), 3.40-3.28 (m, 4H), 3.22-3.12 (m, 1H), 2.38-2.30 (m, 1H), 2.25-2.15 (m, 1H), 2.04- 1.43 (m, 15 H), 1.29-0.96 (m, 12 H), 0.78 (s, 6 H); ¹³ C NMR (100 MHz, CDCl ₃ ): δ 140.6, 121.7, 109.4, 80.9, 79.8, 70.9, 69.7, 66.9, 64.2, 62.2, 56.6, 50.2, 41.7, 40.4, 39.8, 39.1, 37.2, 37.1, 32.2, 31.9, 31.5, 30.4, 28.9, 28.4, 20.9, 19.5, 17.2, 16.4, 14.6.

<17-2> SMA - Of D1a  synthesis

Compound SMA-Dla was synthesized in 85% yield according to the general saccharification reaction procedure of Example 3-2. ¹ H NMR (400 MHz, CDCl ₃ ): δ 8.18-7.96 (m, 10H), 7.90-7.64 (m, 18H), 7.53-7.10 (m, 42H), 6.23-6.12 (m, 2H), 5.86- 5.62 (m, 8H), 5.35-5.23 (m, 4H), 5.12-4.68 (m, 6H), 4.59-4.27 (m, 13H), 4.10 (q, J = 7.6 Hz, 1H), 3.95-3.82 ( m, 2H), 3.50-3.35 (m, 4H), 3.05-2.96 (m, 1H), 1.96-1.88 (m, 2H), 1.80-1.25 (m, 15H), 1.22-0.80 (m, 12H), 0.77 (s, 6 H); ¹³ C NMR (100 MHz, CDCl ₃ ): δ 166.2, 166.1, 165.9, 165.8, 165.7, 165.6, 165.5, 165.1, 165.0, 133.5, 133.3, 130.0, 129.9, 129.8, 129.7, 129.6, 129.5, 129.4, 129.2, 128.9, 128.8, 128.7, 128.6, 128.5, 128.4, 128.3, 128.2, 128.0, 109.4, 96.5, 80.9, 73.3, 72.8, 72.4, 70.9, 70.1, 69.2, 69.1, 68.2, 66.9, 63.5, 62.6, 62.1, 56.5, 50.0, 41.6, 40.3, 36.8, 36.7, 32.0, 31.8, 30.3, 19.3, 17.2, 16.3, 14.6. 14.1.

<17-3> SMA Synthesis of -D1

SMA-D1 in 94% yield according to the general synthetic procedure for deprotection reaction of Example 3-3 Synthesized. ¹ H NMR (400 MHz, CD ₃ OD): δ 5.37 (d, J = 4.0 Hz, 1H), 5.17 (d, J = 4.0 Hz, 2H), 4.52-4.46 (m, 1H), 4.42-4.36 ( m, 2H), 4.07-4.01 (m, 2H), 3.92-3.77 (m, 8H), 3.70-3.62 (m, 10H), 3.53-3.40 (m, 5H), 3.30-3.18 (m, 4H), 2.42-2.34 (m, 1H), 2.22-2.16 (m, 1H), 2.03-1.88 (m, 5H), 1.74-1.40 (m, 12H), 1.19 (t, J = 7.6 Hz, 4H), 1.03 ( s, 4H), 0.97 (d, J = 7.6 Hz, 4H), 0.81 (s, 6H); ¹³ C NMR (100 MHz, CD ₃ OD): δ 142.07, 122.7, 122.6, 110.6, 104.8, 104.5, 104.3, 104.2, 102.9, 98.2, 93.9, 82.3, 81.9, 81.5, 81.3, 81.2, 81.1, 77.9, 77.8 , 77.7, 76.7, 76.6, 75.9, 75.2, 75.1, 74.9, 74.8, 74.7, 74.3, 74.2, 73.5, 71.7, 71.6, 71.5, 67.9, 66.9, 63.8, 62.8, 62.4, 62.3, 57.9, 51.7, 43.0, 41.5 , 41.0, 40.2, 38.5, 38.2, 33.3, 32.9, 32.5, 31.5, 30.1, 29.5, 29.4, 22.1, 20.1, 17.7, 16.9, 15.6, 15.1; HRMS ( FAB ⁺ ) : calcd. for C ₅₄ H ₈₈ O ₂₅ [M + Na] ⁺ 1159.5512, observed 1159.5515.

< Production Example  18> SMA Synthesis of -D2

<18-1> Synthesis of Compound C2

Compound C2 was synthesized in an 80% yield according to the synthesis procedure of Example 3-1-2. ¹ H NMR (400 MHz, CDCl ₃ ): δ 5.33 (d, J = 4.0 Hz, 1H), 4.42 (q, J = 7.6 Hz, 1H), 3.84 (br s, 1H), 3.72-3.68 (m, 1H), 3.64-3.45 (m, 4H), 3.40-3.28 (m, 4H), 3.22-3.12 (m, 1H), 2.38-2.30 (m, 1H), 2.25-2.15 (m, 1H), 2.06- 1.43 (m, 17 H), 1.29-0.96 (m, 12 H), 0.79 (s, 6H); ¹³ C NMR (100 MHz, CDCl ₃ ): δ 140.7, 121.6, 109.4, 80.9, 79.5, 71.6, 66.9, 66.5, 65.9, 62.2, 56.6, 50.2, 41.7, 40.4, 39.9, 39.1, 39.0, 37.2, 37.1, 33.2, 32.2, 31.9, 31.5, 31.4, 30.4, 28.9, 28.5, 28.4, 20.9, 19.5, 17.3, 16.4, 14.6.

<18-2> SMA - Of D2a  synthesis

Compound SMA-D2a was synthesized in a yield of 85% according to the general saccharification reaction procedure of Example 3-2. ¹ H NMR (400 MHz, CDCl ₃ ): δ 8.16-7.96 (m, 10H), 7.90-7.64 (m, 18H), 7.53-7.10 (m, 42H), 6.22-6.12 (m, 2H), 5.86- 5.62 (m, 8H), 5.35-5.23 (m, 4H), 5.12-4.68 (m, 6H), 4.59-4.27 (m, 13H), 4.10 (q, J = 7.6 Hz, 1H), 3.95-3.82 ( m, 2H), 3.50-3.35 (m, 4H), 3.05-2.96 (m, 1H), 1.96-1.88 (m, 2H), 1.80-1.25 (m, 17H), 1.22-0.80 (m, 12H), 0.78 (s, 6 H); ¹³ C NMR (100 MHz, CDCl ₃ ): δ 166.3, 166.1, 165.9, 165.8, 165.7, 165.6, 165.5, 165.1, 165.0, 133.5, 133.3, 130.0, 129.9, 129.8, 129.7, 129.6, 129.5, 129.4, 129.2, 128.9, 128.8, 128.7, 128.6, 128.5, 128.4, 128.3, 128.2, 128.0, 109.4, 96.5, 80.9, 73.3, 72.8, 72.4, 70.9, 70.1, 69.2, 69.1, 68.2, 66.9, 63.5, 62.6, 62.1, 56.5, 50.0, 41.6, 40.3, 36.8, 36.7, 32.0, 31.7, 30.2, 19.3, 17.2, 16.3, 14.6. 14.2.

<18-3> SMA Synthesis of -D2

SMA-D2 in 94% yield according to the general synthetic procedure for deprotection reaction of Example 3-3 Synthesized. ¹ H NMR (400 MHz, CD ₃ OD): δ 5.37 (d, J = 4.0 Hz, 1H), 5.17 (d, J = 4.0 Hz, 2H), 4.49 (d, J = 8.0 Hz, 1H), 4.42 -4.36 (m, 2H), 4.07-4.01 (m, 2H), 3.96-3.76 (m, 8H), 3.70-3.62 (m, 10H), 3.53-3.40 (m, 5H), 3.30-3.18 (m, 4H), 2.42-2.34 (m, 1H), 2.22-2.16 (m, 1H), 2.03-1.88 (m, 5H), 1.74-1.40 (m, 14H), 1.19 (t, J = 7.6 Hz, 4H) , 1.03 (s, 4H), 0.97 (d, J = 7.6 Hz, 4H), 0.80 (s, 6H); ¹³ C NMR (100 MHz, CD ₃ OD): δ 142.2, 122.6, 110.6, 104.5, 102.9, 99.5, 98.2, 93.9, 82.3, 81.8, 81.4, 81.3, 81.2, 80.6, 77.9, 77.8, 77.7, 76.7, 75.9 , 75.1, 74.8, 74.3, 74.2, 73.5, 71.7, 71.6, 71.5, 68.5, 67.9, 63.8, 62.8, 62.4, 62.3, 57.9, 51.7, 50.0, 43.0, 41.5, 41.02, 40.3, 38.5, 38.2, 33.3, 32.9 , 32.5, 31.5, 29.9, 29.6, 22.1, 20.0, 17.6, 16.9, 15.1; HRMS (FAB ⁺ ) : calcd. for C ₅₅ H ₉₀ 0 ₂₅ [M + H] ⁺ 1151.5849, observed 1151.5846.

< Production Example  19> SMA Synthesis of -D3

<19-1> Synthesis of Compound C3

Compound C3 was synthesized in an 80% yield according to the synthesis procedure of Example 3-1-2. ¹ H NMR (400 MHz, CDCl ₃ ): δ 5.33 (d, J = 4.0 Hz, 1H), 4.42 (q, J = 7.6 Hz, 1H), 3.84 (br s, 1H), 3.72-3.68 (m, 1H), 3.64-3.45 (m, 4H), 3.40-3.28 (m, 4H), 3.22-3.12 (m, 1H), 2.38-2.30 (m, 1H), 2.25-2.15 (m, 1H), 2.06- 1.43 (m, 19 H), 1.29-0.96 (m, 12 H), 0.79 (s, 6H); ¹³ C NMR (100 MHz, CDCl ₃ ): δ 140.8, 121.6, 109.4, 80.9, 79.5, 71.6, 66.9, 66.5, 65.9, 62.2, 56.6, 50.2, 41.7, 40.4, 39.9, 39.1, 39.0, 37.2, 37.1, 33.2, 32.2, 31.9, 31.5, 31.4, 30.4, 28.9, 28.5, 26.7, 28.4, 21.2, 21.0, 19.5, 17.3, 16.4, 14.6.

<19-2> SMA - Of D3a  synthesis

Compound SMA-D3a was synthesized in a yield of 85% according to the general saccharification reaction procedure of Example 3-2. ¹ H NMR (400 MHz, CDCl ₃ ): δ 8.18-7.96 (m, 10H), 7.90-7.64 (m, 18H), 7.53-7.10 (m, 42H), 6.23-6.10 (m, 2H), 5.86- 5.62 (m, 8H), 5.35-5.23 (m, 4H), 5.12-4.66 (m, 6H), 4.59-4.27 (m, 13H), 4.10 (q, J = 7.6 Hz, 1H), 3.95-3.82 ( m, 2H), 3.50-3.35 (m, 4H), 3.05-2.96 (m, 1H), 1.96-1.88 (m, 2H), 1.82-1.25 (m, 19H), 1.22-0.80 (m, 12H), 0.77 (s, 6 H); ¹³ C NMR (100 MHz, CDCl ₃ ): δ 166.2, 166.1, 165.9, 165.8, 165.7, 165.6, 165.5, 165.1, 165.0, 133.5, 133.3, 130.0, 129.9, 129.8, 129.7, 129.6, 129.5, 129.4, 129.2, 128.9, 128.8, 128.7, 128.6, 128.5, 128.4, 128.3, 128.2, 128.0, 109.4, 96.5, 80.9, 73.3, 72.8, 72.4, 70.9, 70.1, 69.2, 69.1, 68.2, 66.9, 63.5, 62.6, 62.1, 56.5, 50.0, 41.6, 40.3, 39.8, 36.8, 36.7, 32.0, 31.8, 30.3, 19.3, 17.2, 16.3, 14.6. 14.3.

<19-3> SMA Synthesis of -D3

SMA-D3 at 94% yield according to the general synthetic procedure for deprotection reaction of Example 3-3 Synthesized. ¹ H NMR (400 MHz, CD ₃ OD): δ 5.37 (d, J = 4.0 Hz, 1H), 5.17 (d, J = 4.0 Hz, 2H), 4.48 (d, J = 8.0 Hz, 1H), 4.42 -4.35 (m, 2H), 4.07-4.01 (m, 2H), 3.96-3.76 (m, 8H), 3.70-3.62 (m, 10H), 3.53-3.40 (m, 5H), 3.30-3.18 (m, 4H), 2.42-2.35 (m, 1H), 2.22-2.16 (m, 1H), 2.03-1.88 (m, 5H), 1.74-1.40 (m, 16H), 1.19 (t, J = 7.6 Hz, 4H) , 1.03 (s, 4H), 0.97 (d, J = 7.6 Hz, 4H), 0.79 (s, 6H); ¹³ C NMR (100 MHz, CD ₃ OD): δ 142.2, 122.6, 110.7, 104.6, 104.3, 103.0, 98.3, 93.9, 82.3, 81.9, 81.5, 77.9, 77.8, 76.8, 76.6, 75.9, 75.2, 74.8, 74.4 , 74.2, 73.6, 71.7, 71.6, 67.9, 67.0, 63.8, 62.8, 62.5, 62.4, 62.3, 57.9, 51.8, 43.0, 41.5, 41.0, 40.3, 38.5, 38.2, 33.3, 32.9, 32.8, 32.6, 31.5, 30.0 , 29.6, 26.9, 22.1, 20.0, 17.6, 16.9, 15.5, 15.1; HRMS ( FAB ⁺ ) : calcd. for C ₅₆ H ₉₂ O ₂₅ [M + Na] ⁺ 1187.5825, observed 1187.5831.

< Experimental Example  1> GDNs , SPSs , SPS - Ls  And SMAs  Characteristics of

In order to confirm the properties of GDNs, SPSs, SPS-Ls and SMAs of Preparation Examples 1 to 19 synthesized according to the synthesis methods of Examples 1 to 3, the molecular weight (MW) of GDNs, SPSs, SPS-Ls and SMAs, Critical micellar concentration (CMC) and hydrodynamic radii ( R _h ) of the micelles formed were measured.

Specifically, the critical micelle concentration (CMC) was measured using fluorescence staining, diphenylhexatriene (DPH), and the hydrodynamic radius ( R _h ) of micelles formed by each agent (1.0 wt%) was determined. It was measured through dynamic light scattering (DLS) experiments. The measured results are shown in Table 1 in comparison with the existing amphiphilic molecules (DDM).

MW CMC (μM) CMC (wt%) R _h (nm) ^a GDN-1 1081.2 To 330 ~ 0.036 5.4 ± 0.2 GDN-2 1081.2 To 270 ~ 0.029 6.3 ± 0.5 SPS-1 1199.4 ~ 2 ~ 0.0002 3.4 ± 0.1 SPS-2 1197.4 ~ 3 ~ 0.0003 3.4 ± 0.2 SPS-3 1225.4 ~ 2 ~ 0.0002 3.5 ± 0.1 SPS-4 1225.3 To 150 ~ 0.018 3.1 ± 0.2 SPS-1L 1257.5 To 1 ~ 0.0001 4.1 ± 0.1 SPS-2L 1255.5 ~ 3 ~ 0.0004 4.9 ± 0.1 SPS-3L 1283.5 ~ 0.8 ~ 0.0001 4.9 ± 0.1 SPS-4L 1283.4 To 200 ~ 0.026 3.9 ± 0.0 GDN 1165.3 To 18 ~ 0.0021 3.9 ± 0.1 SMA-A1 1111.32 ~ 4 ~ 0.0004 3.8 ± 0.06 SMA-A2 1125.35 ~ 2 ~ 0.0002 4.0 ± 0.15 SMA-A3 1139.38 ~ 0.8 ~ 0.00009 5.7 ± 0.16 SMA-E1 1109.31 ~ 2 ~ 0.0002 4.3 ± 0.19 SMA-E3 1137.36 ~ 0.7 ~ 0.00008 6.5 ± 0.12 SMA-D1 1137.27 To 20 ~ 0.0023 3.9 ± 0.14 SMA-D2 1151.30 To 12 ~ 0.0014 3.9 ± 0.12 SMA-D3 1165.33 ~ 4 ~ 0.0005 4.2 ± 0.04 DDM 510.62 To 170 ~ 0.0087 3.4 ± 0.03

^a Molecular weight of detergents. ^b Detergent hydrodynamic radius measured at 0.5 wt% detergent concentration by dynamic light scattering (DLS).

The CMC value (0.0008-0.33 mM) of most GDNs, SPSs, SPS-Ls or SMAs was significantly smaller compared to the CMC value (0.17 mM) of DDM. Therefore, micelles are easily formed even at low concentrations of GDNs, SPSs, SPS-Ls, or SMAs, and thus may exhibit the same or superior effect even when using less than DDM.

Meanwhile, as a result of examining the size distribution of micelles formed by GDNs, SPSs, SPS-Ls or SMAs through DLS, GDNs, SPSs, SPS-Ls or SMAs showed micelles of only one population, which means that micelle homogeneity High (FIGS. 4-6).

From these results, most GDNs, SPSs, SPS-Ls or SMAs of the present invention have a lower CMC value than DDM, so that micelles are easily formed in a small amount, so that the self-assembly tendency is much higher than that of DDM. The micelles were confirmed to be homogeneous.

< Experimental Example  2> GDNs , SPSs , SPS - Ls  And SMAs  of LeuT Membrane protein  Evaluation of Structural Stabilization Capability

Thermophilic bacteria Aquifex Aquifex aeolicus ) derived wild type LeuT (leucine transporter) was purified by the method described previously ( 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 8xHis-tagged transporter (expression plasmids were provided by Dr E. Gouaux, Vollum Institute, Portland, Oregon, USA) . In summary, separation of the bacterial membrane, and 1% (w / v) after solubilization in DDM, and coupling a protein to the Ni ^{2 +} -NTA resin (Life Technologies, Denmark), 20 mM Tris-HCl (pH 8.0), 1mM Elution was in NaCl, 199 mM KCl, 0.05% (w / v) DDM and 300 mM imidazole. Thereafter, purified LeuT (about 1.5 mg / ml), except for DDM and imidazole, in buffers equivalent to the above, DDM (control), GDNs (GDN, GDN-1, or GDN-2), SPSs (SPS- 1, SPS-2, SPS-3 or SPS-4), SPS-Ls (SPS-1L, SPS-2L, SPS-3L or SPS-4L) or SMAs (SMA-As, SMA-Es or SMA-Ds) Diluted 10-fold with buffer supplemented to this final concentration CMC + 0.04 wt%. Protein samples were incubated for 10 days at room temperature, centrifuged at regular intervals during incubation, and protein properties were confirmed by measuring [ ³ H] -Leucine binding capacity using SPA. SPA was performed with 5 μL of protein samples in a buffer of the above-mentioned concentrations containing 450 mM NaCl and GDNs, SPSs or SPS-Ls (or DDM). SPA reactions were performed in the presence of 20 nM [ ³ H] -Leucine and 1.25 mg / ml copper chelate (His-Tag) YSi beads (Perkin Elmer, Denmark). Total [ ³ H] -Leucine binding for each sample was measured using a MicroBeta liquid scintillation counter (Perkin Elmer).

As shown in Figures 8 and 9, the initial substrate binding activity of LeuT dissolved in DDM is superior to other amphiphilic compounds of the present invention, but it was confirmed that the activity rapidly deteriorated over time. As a result of comparing the long-term substrate binding activity, it was confirmed that most of SMAs except SMA-A3 and SMA-D3 had a significantly better effect of maintaining the substrate binding properties of LeuT than DDM during the incubation period of 12 days or more.

₂ AR 막단백질 구조 안정화 능력 평가 <Experiment 3> GDNs, SPSs, SPS - Ls and the SMAs

₂ AR Evaluation of membrane protein structure stabilization ability

₂ 아드레날린성 수용체 (

₂AR), G-단백질 연결 수용체(GPCR) 구조 안정성을 측정하는 실험을 하였다. 즉, DDM으로 정제된 수용체는 CHS (cholesteryl hemisuccinate) 없이 각각의 GDNs, SPSs, SPS-Ls 및 SMAs만을 함유하는 버퍼 용액 또는 CHS와 DDM을 함유하는 버퍼 용액으로 희석시켰다. 최종 화합물 농도는 CMC+0.2 wt%이었으며, 수용체의 리간드 결합 특성은 [³H]-디하이드로알프레놀올 ([³H]-DHA)의 결합에 의해 측정하였다.Human by GDNs, SPSs, SPS-Ls and SMAs

₂ adrenergic receptors (

₂ AR), G-protein coupled receptor (GPCR) structural stability experiments were measured. That is, the receptor purified with DDM was diluted with a buffer solution containing only GDNs, SPSs, SPS-Ls and SMAs, or a buffer solution containing CHS and DDM, without cholesteryl hemisuccinate (CHS). Final compound concentrations were CMC + 0.2 wt%, the ligand binding characteristics of the receptor is ^[3 H] - were measured by a combination of dihydro alpeure nolol ^([3 H] -DHA).

₂AR는 0.1% DDM을 사용하여 정제하였으며 (D. M. Rosenbaum 등, Science, 2007, 318, 1266-1273.), 약 10 mg/ml (약 200 μM)로 최종 농축하였다. DDM으로 정제된

₂AR를 사용하여 0.2% 양친매성 화합물 (DDM, GDNs, SPSs, SPS-Ls 및 SMAs)에서 0.5 mg/ml BSA로 보충된 10 nM [³H]-Dihydroalprenolol (DHA)를 함유하는 마스터 결합 혼합물을 제조하였다. DDM, GDNs, SPSs, SPS-Ls 및 SMAs로 정제된 수용체는 상온에서 30분 동안 10 nM의 [³H]-DHA와 함께 인큐베이션하였다. 혼합물을 G-50 컬럼에 로딩하고, 통과액을 1 ml 바인딩 버퍼 (0.5 mg/ml BSA로 보충된 20 mM HEPES pH 7.5, 100 mM NaCl)로 수집하고, 그리고 15 ml 섬광 유체(scintillation fluid)로 채웠다. 수용체-결합된 [³H]-DHA는 섬광 카운터 (Beckman)로 측정했다. [³H]-DHA의 결합도는 컬럼 그래프로 나타내었다.Specifically, the radioligand binding test used the following method.

₂ AR was purified using 0.1% DDM (DM Rosenbaum et al., Science , 2007, 318, 1266-1273.) And finally concentrated to about 10 mg / ml (about 200 μM). Purified with DDM

₂ AR was used to prepare a master binding mixture containing 10 nM [ ³ H] -Dihydroalprenolol (DHA) supplemented with 0.5 mg / ml BSA in 0.2% amphipathic compounds (DDM, GDNs, SPSs, SPS-Ls and SMAs). Prepared. Receptors purified with DDM, GDNs, SPSs, SPS-Ls and SMAs were incubated with 10 nM of [ ³ H] -DHA at room temperature for 30 minutes. The mixture was loaded on a G-50 column, the flow through with 1 ml binding buffer (20 mM HEPES pH 7.5, 100 mM NaCl supplemented with 0.5 mg / ml BSA), and with 15 ml scintillation fluid. Filled. Receptor-bound [ ³ H] -DHA was measured with a scintillation counter (Beckman). The degree of binding of [ ³ H] -DHA is shown in a column graph.

As shown in Figures 10 and 11, most GDNs, SPSs, SPS-Ls and SMAs showed similar or superior effects to DDM in terms of initial activity in maintaining ligand binding properties of the receptor. GDNs and SPS-Ls showed significantly better effects than DDM in long-term receptor ligand binding properties, and in particular, the effect of SPS-1L was excellent.

< Example  4> GDNs , SPSs , SPS - Ls  And Of SMAs MelB _st Membrane protein  Evaluation of Structural Stabilization Capability

Experiments were conducted to determine the structural stability of SalBella typhimurium melibiose permease (MelB _St ) protein by GDNs, SPSs, SPS-Ls and SMAs.

Specifically, wild type MelBSt (melibiose permease) derived from Salmonella typhimurium having a 10-His tag at the C-terminus using the plasmid pK95ΔAHB / WT MelBSt / CH10 was used as an E. coli DW2 strain (ΔmelB). And ΔlacZY). Cell growth and membrane preparation were performed according to the method described in AS Ethayathulla et al. (Nat. Commun. 2014, 5, 3009). Protein assays were performed with the Micro BCA kit (Thermo Scientific, Rockford, IL). Membrane samples containing MelB _St (final protein concentration was 10 mg / mL ^{- 1} ) in solubilization buffer (50 mM sodium phosphate, pH 7.5, 200 mM NaCl, 10% glycerol, 20 mM melibiose) were added to 1.5 wt% DDM, Mixed with GDNs, SPSs, SPS-Ls or SMAs. The resulting sample was incubated at 23 ° C. for 90 minutes. To remove insoluble materials, Beckman Optima® equipped with TLA-100 rotor at 4 ° C for 30 minutes. The ultracentrifugation was performed at 355,590 g with a MAX ultracentrifuge. 20 μg of non-centrifuged membrane protein was applied to the same amount of extract of the compounds after untreated membrane or ultracentrifugation, and the treated samples were loaded into each well in equal volumes. The loaded sample was analyzed by SDS-15% PAGE and then visualized by immunoblotting with Penta-His-HRP antibody (Qiagen, Germantown, MD). To evaluate the thermal stability of MelB _St in a novel amphiphilic compound, membrane extracts extracted at 23 ° C. with individual compounds were further heat treated at 3 different temperatures (65, 70 and 75 ° C.) for an additional 90 minutes, followed by ultracentrifugation followed by SDS- Analysis was by 15% PAGE and Western blotting. MelB _St was detected using a SuperSignal West Pico chemiluminescent substrate by ImageQuant LAS 4000 Biomolecular Imager (GE Healthcare Life Science).

As shown in Figures 12 to 14, at relatively low temperatures of 23 ° C, DDM shows a similar or better effect than other novel compounds, but at high temperatures above 65 ° C, most GDNs, SPSs, SPS-Ls and SMAs are Excellent MelB _st solubilization ability.

In particular, it was confirmed that at high temperatures (above 55 ° C), DDM does not solubilize MelB _st at all, while most GDNs, SPSs, SPS-Ls and SMAs maintain MelB _st solubilization ability even at 55 ° C or higher.

In addition, SPS-3 and SPS-3L, compared to DDM, not only MelB _St, but also MelB _Ec , which is a less stable homologue, was found to have superior protein solubility and ability to maintain the function of the membrane protein. (FIG. 12 (b)).

Claims (18)

A compound represented by the formula of any one of formulas (1) to (3):
[Formula 1]

[Formula 2]

[Formula 3]

In Chemical Formulas 1 to 3,
Dotted line in the formula represents a single bond or a double bond,
L ¹ is a direct bond,

or

N is an integer of 1, 2, 3, 4 or 5, and X ⁴ is a saccharide linked to oxygen;
X ² and X ³ are each independently a saccharide connected with hydrogen or oxygen, and
X ¹ is a saccharide connected with oxygen.
The compound of claim 1, wherein the saccharide is a monosaccharide, a disaccharide, or a pentasaccharide.
The compound of claim 1, wherein the saccharide is glucose, maltose or a branched pentasaccharide of glucose centers.
The compound of claim 1, wherein L ¹ is a direct bond; One of X ² and X ³ is hydrogen and the other is a saccharide linked with oxygen; And wherein X ¹ is a saccharide linked with oxygen.
The method of claim 1, wherein L ¹

N is an integer of 1, 2, 3, 4 or 5; X ² and X ³ are hydrogen; And wherein X ¹ is a saccharide linked with oxygen.
The method of claim 1, wherein L ¹

N is an integer equal to 1, 2, 3, 4 or 5, and X ⁴ is a saccharide linked to oxygen; X ² and X ³ are hydrogen; And wherein X ¹ is a saccharide linked with oxygen.
The compound of claim 1, wherein the compound is one of Formulas 4 to 22:
[Formula 4]

[Formula 5]

[Formula 6]

[Formula 7]

[Formula 8]

[Formula 9]

[Formula 10]

[Formula 11]

[Formula 12]

[Formula 13]

[Formula 14]

[Formula 15]

[Formula 16]

[Formula 17]

[Formula 18]

[Formula 19]

[Formula 20]

[Formula 21]

[Formula 22]

The compound of claim 1, wherein the compound has a critical micelle concentration (CMC) in an aqueous solution of 0.0001 to 1 mM.
Composition for solubilizing the membrane protein containing the compound according to claim 1.
Composition for extracting membrane protein comprising the compound according to claim 1.
A composition for stabilizing a membrane protein comprising the compound according to claim 1.
A composition for crystallization of a membrane protein comprising the compound according to claim 1.
Composition for the analysis of membrane proteins comprising a compound according to claim 1.
The composition of claim 9, wherein the composition is a formulation of micelles, liposomes, emulsions or nanoparticles.
The method according to any one of claims 9 to 13, wherein the membrane protein is LHI-RC, LeuT (Leucine transporter), β ₂ AR (human β ₂ adrenergic receptor), MelB _st (melibiose permease) or two or more thereof Composition in combination.
1) Cis or trans hydrides on carbons 2 and 3 of the steroids of ticogenin, diosgenin, cholesterol, Cholesterol, Cholestanol or Sitosterol Introducing oxy;
2) introducing a saccharide to which a protecting group is attached by performing a glycosylation reaction on the product of step 1); And
3) performing a deprotection reaction on the product of step 2); a method of preparing a compound represented by one of the following Chemical Formulas 1 to 3:
[Formula 1]

[Formula 2]

[Formula 3]

In Chemical Formulas 1 to 3,
Dotted line in the formula represents a single bond or a double bond,
L ¹ is a direct bond;
X ¹ and X ² are sugars connected with oxygen, and X ³ is hydrogen.
1) introducing hydroxy to carbon 3 and 6 of the steroid of ticogenin, diosgenin, cholesterol (Cholesterol), cholestanol (Cholestanol) or cytosterol (Sitosterol);
2) introducing a saccharide to which a protecting group is attached by performing a glycosylation reaction on the product of step 1); And
3) performing a deprotection reaction on the product of step 2); a method for preparing a compound represented by one of the following Chemical Formulas 1 to 3:
[Formula 1]

[Formula 2]

[Formula 3]

In Chemical Formulas 1 to 3,
Dotted line in the formula represents a single bond or a double bond,
L ¹ is a direct bond;
X ¹ and X ³ are saccharides connected with oxygen, and X ² is hydrogen.
1) Introducing a substituted or unsubstituted alkylene linker to the hydroxy of carbon 3 of the steroid of ticogenin, diosgenin, cholesterol, Cholesterol or Citostanol Or performing a glycosylation reaction to introduce a saccharide to which a protecting group is attached;
2) introducing a saccharide to which a protecting group is attached by performing a glycosylation reaction on the product into which the alkylene group linker of step 1) is introduced; And
3) performing a deprotection reaction on the glycosylation product of step 1) or the product of step 2); Method for producing a compound represented by any one of formulas 1 to 3, including:
[Formula 1]

[Formula 2]

[Formula 3]

In Chemical Formulas 1 to 3,
L ¹ is a direct bond,

or

N is an integer of 1, 2, 3, 4 or 5, and X ⁴ is a saccharide linked to oxygen;
X ² and X ³ are hydrogen; And
X ¹ is a saccharide connected with oxygen.

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