WO2015084075A1 - Cucurbituril-sugar conjugate and method for preparing same - Google Patents

Abstract

The present invention provides a cucurbituril-sugar conjugate and a method for preparing the same, wherein cucurbituril recognizes sugar to support and stabilize the sugar in a cucurbituril cavity.

Description

 Specification

 NAME OF THE INVENTION: Cooker bituril-sugar complexes and methods for making the same

 [1] The present invention relates to a cooker bituril-sugar complex and a method for producing the same.

Details of the process are related to cooker bituril [ ^η ] -sugar complexes in which the cooker bitu [n] reel recognizes sugars and co-supports the sugars and stabilizes them.

 Background

 [2] Artificial saccharide receptors can be used for applications such as sensing and drug delivery. Artificial saccharide receptors, which act on the basis of non-covalent bonds in aqueous solutions, have emerged as very interesting topics in biochemistry or supramolecular chemistry. have.

 [3] In the implementation of these saccharide receptors, the first and the most difficult is the separation of sugars and water molecules and the subsequent association of selective saccharides. Unlike covalent bond-based boronic acids, saccharide phosphoproteins There is only a handful of artificial waters with functional groups capable of hydrogen bonding with hydrophobic cages, such as lectins.

 [4] Unlike organic solvent-based receptors, they are also structurally

 There is a limit to the implementation of receptors that recognize functionally diverse sugars.

 [5] Aminosaccharides, on the surface of the cell, are mostly present in the form of N-acylated compounds, which serve as biomarkers for many biological processes and diseases. They are also known as aminoglycosides. It is also an important factor.

 [6] However, despite the potential applicability of artificial receptors, there are still many difficulties in using them in recognition of aminosaccharides.

 [7] As mentioned above, organic solvent-based receptors are mounded, and artificial receptors that act on aqueous solutions based on non-covalent bonds have not been realized.

 [8] Some boric acid-based receptors exist, but they have very weak bonds and metal ions are also recognized as aminosaccharides, but their mating is limited to detection, so that they are based on non-covalent bonds in aqueous solutions. Research is highly required.

 Detailed description of the invention

 Technical task

 [9] The present invention detects artificial lectins for glycan analysis and detects cell-cell interactions. Lipid-CB [7] Cookerbituril useful for the transport and detection of conjugates and antibody drugs. Provided are the complexes and methods for their preparation.

Task solution [10] The present invention provides a cooker bituryl-sugar complex formed by supramolecular chain cooker bituril which recognizes and supports saccharides in an aqueous solution.

 [11] cooker bituril,

[12] Contains sugars supported in the joint of the cooker bituril.

The cooker bituril according to an embodiment of the present invention may be represented by Chemical Formula 1.

 [14] [Formula 1]

 [16] [In Formula 1,

[17] X is 0, S or NH,

[18] A, and八₂ are each independently selected from H, OR, SR, NHR, COOH, 0 (CH _{2) a} R or 0 (CH _{2) a} SR, wherein a is 1 to 5 can Protocol, the R is H, halogen, C1-C30 alkyl, C2-C30 alkenyl, C2-C30 alkynyl, C2-C30 carbonylalkyl, C3-C30 cycloalkyl, C2-C30 heterocycloalkyl, C6-C30 aryl, C6- C30 aryl CI-C30 alkyl ，

 C2-C30 heteroaryl or C2-C30 heteroaryl C1-C30 alkyl;

 [19] n is an integer between 4 and 14.

Preferably, in Formula 1 according to one embodiment of the present invention X is 0, A, and A 2 are independently H, OR, 0 (CH ₂ ) aR or 0 (CH ₂ ) aSR, A is an integer of 1 to 5, and R is H, C1-C30 alkyl or C2-C30 alkenyl; n can be an integer from 6 to 10.

 According to one embodiment of the present invention, the sugar may be a monosaccharide, a disaccharide, or a polysaccharide, and preferably a sugar having an amino group or a salt containing the same.

 Preferably, a sugar having an amino group or a salt containing the same according to an embodiment of the present invention may be represented by an amino sugar antibiotic or the following formula (2).

[23] [Formula 2]

 [25] [In Formula 2,

[26] R _n to R _l5 are independently of each other OH, NH ₂ or NH ₃ ⁺ X—, and any one of R _n to R _l5 is necessarily NH ₂ or NH ₃ ⁺ X-; [27] X- is a monovalent ion of acid.]

 The cooker bituril fructose according to one embodiment of the present invention may have an equivalent ratio of 0.5 to 3: 1.

 [29] The present invention also provides a method for producing a cucurbituryl-sugar complex in which the cucurbituryl sugar is selectively recognized, supported and stabilized in an aqueous solution.

 And contacting the cooker bituril and the sugar in an aqueous solution to produce the cooker bituril-sugar complex.

 [31] A cooker bituril of the present invention is prepared according to one embodiment of the method for preparing a sugar complex.

The cucurbituril may be represented by Chemical Formula 1, and in Chemical Formula 1, X is 0, A, and ^ are each independently H, OR, 0 (CH ₂ ) aR or 0 (CH ₂ ) aSR, a is an integer of 1 to 5, wherein R is H, C1-C30 alkyl or

 C2-C30alkenyl; n can be an integer from 6 to 10.

 [32] The sugar according to one embodiment of the method for preparing the cookie bituril of the present invention

 It may be a sugar having an amino group or a salt containing the same, and specifically, may be represented by an amino sugar antibiotic or the above Chemical Formula 2.

According to one embodiment of the manufacturing method of the composite per ^- [33] of the present invention cucurbituril

 Cooker beryl fructose can have an equivalent ratio of 0.5 to 3: 1.

 [34] The present invention also provides a method for detecting sugars using cooker bituril. Effects of the Invention

 [35] The cooker bituril of the present invention-the sugar complex of the present invention, in which the cooker bituril selectively recognizes sugars in the aqueous solution, which is supported by the cooker bituril joint and supported by the supported sugars and anomers.

 By stabilizing by preventing the rotation of the light, cooker bituril can support and stabilize specific anomers to form an optional cooker bituryl-sugar complex according to the type of sugar.

 [36] The cucurbituril-glycocomplex of the present invention also introduces functionalization of cucurbituril outward rings to allow the analysis of artificial lectins or cells and cells for glycan analysis.

 Application to the lipid-CB [7] conjugate is also possible.

 [37] The cucurbituril-sugar complex of the present invention can also be used for the transport and detection of specific antibody drugs.

 [38] In addition, the cooker bituril of the present invention cooker bituril can recognize sugars in aqueous solution and can detect cell-cell interactions.

[39] In addition, the production method of the cooker bituril-sugar complex of the present invention can be prepared in aqueous solution.

Cooker bituril can recognize sugars and make stabilized cooker bituryl-sugar complexes by co-operating with cooker bituril joint, and can be applied to cell-cell interactions and transport and detection of antibody by the method of producing cooker bituryl-sugar complex of the present invention. have. Brief description of the drawings

1 is a diagram showing the structure of a cooker bite reel,

2 is a diagram showing amino sugars used in the embodiment of the present invention,

[42] Figure 3 shows CB [7]. This diagram shows the NMR spectrum of one complex.

[43] Fig. 4 is a diagram showing the MALDI-TOF mass spectrum of CB [7] · one composite, [44] Fig. 5 is CB [7]. 1 complex and 1 (bottom) proton-coupled ¹³ C-NMR spectrum

 It is a drawing shown.

 [45] Figure 6 shows the ROESY (278K) of CB [7] .1 complex formed from two equivalents of CB].

 Is a diagram representing the spectrum,

7 is a diagram showing an NMR spectrum of a CB [7] -2 complex.

[47] Fig. 8 is a diagram showing the MALDI-TOF mass spectrum of CB [7] -two complexes. [48] Fig. 9 is CB [7]. 2 complex and 2 (bottom) proton-coupled ¹³ C-NMR spectrum

 The drawing is shown,

 10 is a diagram showing a DQF-COSY spectrum of a CB [7] .complex formed from two equivalents of CB [7].

 [50] Figure 11 shows the ROESY (278K) of CB [7] .2 complex formed from two equivalents of CB [group].

 It is a diagram showing the spectrum,

 [51] Figure 12 is a molecular dynamics modeling of B [7] -two complexes,

 FIG. 13 is a diagram showing an NMR spectrum of 2 when a CB group is added. FIG.

14 shows CB [7] (0.5mM) and 2 (10mM) ITC profiles in water at 298K.

 15 is a diagram showing a 1 H NMR of a CB [7] .3 complex formed from two equivalents of CB [group].

 FIG. 16 shows CB [3 and 10 (mM) ITC profiles in water at 298 K, [56] FIG. 17 shows the Ή-NMR spectrum of 4 when CB [added], [57] FIG. 18 shows a CB [4] and 4 (10 mM) ITC profile in water at 298K. Mode for Carrying Out the Invention

[58] While investigating the anomeric effects of saccharide receptors, the present inventors anticipated that supramolecules could stabilize the alpha anomers by immersing them in hydrophobic cavities in aqueous solutions. To stabilize by knowing, supporting,

 The present invention was completed by preparing a cucurbituril-sugar complex.

[59] The cooker bituril of the present invention—the sugar complex

[60] cooker bituril, and

 [61] The sugar contained in the joint of the cooker bituril is included.

[62] Sugars are usually alpha due to the stereoelectronic effect.

Induces stabilization of the ot-anomer, and the sugars in aqueous state Hydration to beta-anomer

 Induces mutarotation.

 [63] However, the present inventors have found that the artificial acceptor is an ultra-molecular chain cucurbituril (CB [n]).

 The present invention was found to selectively recognize saccharides in aqueous solution, and to stabilize the sugars with alpha or beta-omers by preventing the photochromic phenomena of sugars supported on agacoucurbituril.

 [64] Cookerbitu [n] reel (CB [n]) is a large ring molecule with a hard, hydrophobic cavity inside, which is a supermolecular molecule that dissolves in water without charge and is present in the cavity and inlet of CB [n]. Carbonyl groups enable stable formation of the master-guest complex based on hydrophobic interactions and strong ion-dipole interactions, but no stable cucurbituryl-sugar complexes supported on cucurbituryl sugars have been reported.

 [0065] The cooker bituril of the present invention cooker bituril can be applied to transport and detect aminoglucoside antibody drugs by recognizing and stabilizing sugars in aqueous solution and by co-curing with the cooker bituril joint.

 [66] In addition, the cooker bituril of the present invention can recognize sugars.

 Therefore, a functional group can be introduced into the cucurbituril outbound ring to prepare an artificial lectin for glycan analysis or a lipid-CB [7] conjugate for detecting cell-cell interactions.

 Cooker bituril according to an embodiment of the present invention may be represented by the following formula (1) 68 [68]

[69] 圍^A N-CH _?

 X n

 [70] [In Formula 1,

[71] X is O, S or NH,

A, and A ₂ are each independently H, OR, SR, NHR, COOH, 0 (CH ₂ ) _a R or 0 (CH ₂ ) _a SR, wherein a is an integer of 1 to 5, R is H, halogen, C1-C30 alkyl, C2-C30 alkenyl, C2-C30 alkynyl, C2-C30 carbonylalkyl, C3-C30 cycloalkyl, C2-C30 heterocycloalkyl, C6-C30 aryl, C6- C30 aryl C1-C30 alkyl,

 C2-C30 heteroaryl or C2-C30 heteroaryl C1-C30 alkyl;

[73] _n is an _integer between 4 and 14.

Preferably, in the above Formula 1 according to an embodiment of the present invention X is O, A, and A 2 are independently H, OR, 0 (CH ₂ ) aR or 0 (CH ₂ ) aSR, Where a is 1 to 5 An integer, wherein R is H, C 1 -C 30 alkyl or C 2 -C 30 alkenyl; n could be an integer from 6 to 10.

 More preferably, in Formula 1 according to an embodiment of the present invention, X is 0, and ^ is independently H, OR, and R may be H, C1-C30 alkyl.

 [76] "Alkyl", "alkoxy", and the like described in the present invention, described in the present invention

 Substituents containing an "alkyl" moiety include both straight and branched forms. In addition, "aryl" described in the present invention is aromatic by one hydrogen removal.

Organic radicals derived from hydrocarbons, each comprising a single or fused ring system containing 4 to 7, preferably 5 or 6 ring atoms, each of which has a plurality of aryls connected in a single bond. Include form. Inde a specific example, phenyl, naphthyl, biphenyl, anthryl, carbonyl and the like (indenyl), fluorenyl group, but are not limited to, ^"" heteroaryl "described in the present invention B as aromatic ring backbone atom, 5 to 6 membered ring heteroaryl, which means an aryl group containing 1 to 4 heteroatoms selected from N, 0, S, P (= 0), Si, and P, and the other aromatic ring skeleton atom is carbon. And polycyclic heteroaryls condensed with one or more benzene rings, which may be partially saturated. Heteroaryls in the present invention also include forms in which one or more heteroaryls are linked by a single bond.

 [77] The "cycloalkyl" described in the present invention means a saturated hydrocarbon ring,

Preferably it may be a ₅ to 7 membered alicyclic ring, including when the aromatic or alicyclic ring is fused.

[78] "Heterocycloalkyl" described in the present invention ^. Is a heteroatom containing an oxygen, sulfur or nitrogen in the ring, or a cyclic ring, the number of hetero atoms is 1-4, preferably 1-2. In heterocycloalkyl, cycloalkyl is preferably monocyclo Aryl or alkyl or bicycloalkyl, which is an aromatic ring

 This also includes the case where the heteroaryl is fused.

 [79] C1-C30 alkyl, C2-C30 alkenyl, C2-C30 alkynyl, described in the present invention,

 C2-C30 carbonylalkyl, C3-C30 cycloalkyl and C2-C30 heterocycloalkyl are preferably C1-C10 alkyl, C2-C10 alkenyl, C2-C10 alkynyl, C2-C10 carbonylalkyl, C3-C10 cyclo Alkyl and C2-C10 heterocycloalkyl, and C6-C30 aryl and C4-C30 heteroaryl can be C6-C12 aryl and C4-C12 heteroaryl.

 [80] The sugars and sugars listed in the present invention can be clearly recognized by the person skilled in the art.

 Examples include monosaccharides to polysaccharides, include sugar antibiotics, preferably sugar antibiotics, monosaccharides, disaccharides or polysaccharides, preferably sugars with amino groups or salts containing them, or

 It may be an aminosugar antibiotic, and more preferably an aminosugar antibiotic having an amino group or a salt containing the monosaccharide and an amino group.

[81] Sugar antibiotics and sugar antibiotics of the present invention include sugars based on antibiotics and Antibiotics include antibiotics with or without functional groups and aminosugar antibiotics.

 [82] The aminosugar antibiotics of the present invention are based on sugars having amino groups.

 Antibiotics include, for example, neomycin and ferromycin.

[83] The sugars and amino sugar antibiotics having an amino group or a salt containing the same according to one embodiment of the present invention include, for example, D-glucosamine hydrochloride, D-galactosamine hydrochloride, emannosamine hydrochloride, and ₆ -amino ᅳ 6. -Dioxy-D-glucose, N-acetylglucosamine, sialic acid, L-daunosamine), neomycin and ferromycin, but are not limited to these.

 Specifically, a sugar having an amino group or a salt containing the same according to an embodiment of the present invention may be represented by the following Chemical Formula 2.

[85] [Formula 2]

 [87] [In Formula 2,

R _u to R _l5 are each independently OH, NH ₂ or NH ₃ ⁺ X—, and any one of R _n to R _l5 is NH ₂ or NH ₃ ⁺ X-;

 [89] X- is a monovalent ion of acid.]

[90] cucurbituril and from the side to a strong bond in the general formula (2) in accordance with one embodiment of the invention R "to R ₁₅ are independently a OH or NH ₃ ⁺ X- with each other, to R _l5 is a slow Chinese Language It must be NH ₃ ⁺ X-.

In Formula 2 according to an embodiment of the present invention, X- can be any monoanion of an acid, for example, hydrochloric acid (C1), silver nitrate (N0 _3- ), acetate ions (CH ₃ COO). And preferably hydrochloric acid.

 [92] The cucurbit reel fructose according to one embodiment of the present invention has a molar ratio of 0.5 to 3: 1 days.

 It is preferably 1 to 2: 1.

The present invention also includes a method for producing a cucurbituryl-sugar complex comprising contacting the cucurbituril and a sugar in an aqueous solution to produce a cucurbituryl-sugar complex.

 [94] Specifically, the production of the cooker bituril-sugar complex of the present invention may melt the cooker bituryl fructose in an aqueous solution and contact the cooker bituril fructose, and the mixture of cooker bituryl fructose may be added to the aqueous solution so as to contact the cooker bituril fructose in the aqueous solution. Regardless of whether it is possible.

 [95] According to one embodiment of the method for producing a cookie bituril of the present invention

 Cooked beryl fructose can have an equivalent ratio of 0.5 to 3: 1.

[96] The above-mentioned cooker bituril according to one embodiment of the method for producing a sugar complex of the present invention

b) the reaction time for adding the sugars of step (b) to make the cookiebituryl-sugar complex; The reaction temperature is not limited, but can be performed for 0 to 24 hours at 0 to 80 ^o C.

 [97] According to one embodiment of the method for producing the cookie bituril-sugar complex of the present invention

Cooker bituril may be represented by Formula 1, preferably in Formula 1, X is 0, A, and 八₂ are independently H, OR, 0 (CH ₂ ) aR or 0 (CH ₂ ) aSR, A is an integer of 1 to 5, and R is H, C1-C30 alkyl or

 C2-C30alkenyl; n can be an integer from 6 to 10.

 [98] The sugar according to one embodiment of the method for producing the cookie bituril-sugar complex of the present invention

 It may be a sugar or amino sugar antibiotic having an amino group or a salt containing the same, and specifically, may be represented by Formula 2 above.

 [99] The cooker bituril used in the sugar index can also add functional groups to the outward ring.

 The present invention also provides a method for detecting sugars using cooker bituril.

[101] As mentioned above, the cookerbituril of the present invention recognizes sugars in aqueous solution and stabilizes them by supporting them.

 [102]

 Hereinafter, specific embodiments of the present invention will be described.

 It is not intended to limit the invention to the illustrative purposes of the invention.

 [104] The present invention is the primary molecule that melts in water without charge

 The cucurbitur [girryl (CB [7]) provided a cucurbitur [girryl-sugar complex formed by strong linkages with aminosaccharides; the master-guest interaction was confirmed by NMR spectroscopy and MALDI-TOF mass spectrometry. NMR analysis showed that aminosaccharides exist as alpha anomers in the CB [7] cavity, and ITC analysis showed strong binding to aminosaccharides in CB [gi]. Ka = 1.6 x 10 1 No artificial receptors have been reported (in the case of D-galatosamine hydrochloride) that exhibit strong binding to sugars in aqueous solutions and allow sugar stabilization of certain anomeric forms (particularly alphaanomers).

 Example 1 Preparation of Cookerbitu [D-glucosamine hydrochloride (1) Complex (CB [7] 1 Complex)

 [106] Dissolve 5.8 mg of cucurbituril in 1 mL of water at room temperature.

 Here is the cooker bituril [equal equivalent to the tile or cooker bitur [relative to girill]

 Dissolve 0.5 equivalent of D-glucosamine hydrochloride in 1 mL of water, and add

 Cookerbitu [D-glucosamine hydrochloride (1) complexes were prepared according to their equivalent ratios, respectively.

[107] D-glucosamine hydrochloride (1) is present in excess of alpha anomers when dissolved in D ₂ 0 (α: β = 93: 7) but over time.

After 24 hours due to variable rotation, alpha and beta anomers were present at a ratio of 65:35. X-NMR of the same equivalent amount of CB [7] and 1 D ₂ 0-enzyme solution (5 mM) In the spectrum (Bruker DRX 500 spectrometer, 298 K) we can see the resonance of 1 in the composite (Fig. 3), and through the upfield shift of this signal, 1 binds to the CB [7I side of the hole]. You can see that the wide width of each signal

1 shows a fast transition to the free state with CB [.

 Cookerbitu [The formation of a 1: 1 complex of giryl and 1 was also observed by MALDI-TOF.

 It was confirmed (FIG. 4). The presence of two equivalents of CB [contaminated, thereby fully forming 1 and CB [7] complexes, suppresses fast conversion, resulting in a clear signal of the NMR spectrum (FIG. 3).

[108] Using the 에 and ^l3 C NMR DQF-COZY, HSQC and ROESY spectra (278 and 298 K), the 1 present in the CB ^] 1 complex was fully analyzed. The signal of 1 showed a shift of about 0.6-0.1 ppm and appeared as a set, for example, at 278K, there could be only one anionic peak (4.68 ppm). Has a higher field shift compared to 1 in the free state. The free 1 signals (Η2α (3.2 ppm), Η2β (2 · 9 ppm)) appeared as a single signal in the complex (2.2 ppm), and one double term in the complex's proton coupled ¹³ C-NMR spectrum. doublet) appeared at 88.8 ppm and

 The coupling constant is = 170.4 Hz (Fig. 5). These values are the chemical motion and coupling constants (88.5 ppm,

 = 170.5 Hz), which implies the presence of an alpha anomer in the complex. In addition, in the ROESY spectrum of the complex (Figure 6), Hle-H2 «(l) correlation spot or On the other hand, there is no correlation between Η3α or Η5α with Ηΐα, so we can see that C1-OH of 1 exists in the vertical direction and has the structure of an alpha anomer. Interestingly, CB [Team 1 in ROESY spectrum We see a weak cross-peak between molecules. Comprehensive NMR analysis suggests that CB [stabilizes alpha 1 nanomers.

 [109] Due to the wideness of the signal, the coupling constants of 1 and CB [groups can be

Because of the difficulty in quantitative measurements, the binding constant of complex formation was measured using an isothermal titration calorimetry (ITC). The results of ITC (VP-ITC, Microcal, Inc) (Table 1) showed that CB [strong bonds with Ki 1 (Ka = (4.4 ± 1.0) χ 10 ³ Μ- '). We can see that the changes in the lobby are in the preferred direction (ΔΗ ⁰ = -14.0 KJ mol- ^ TAS ⁰ = 6.6 KJmo) .These changes are discussed below.

[110] Table 1 Table 1

 [111] The standard error was obtained through three independent ITC titration experiments.

[112]

[113] [Example ¾ Cookerbitu [ ⁷ ] Preparation of D-galactosamine hydrochloride (2) Complex (CB [7] 2 Complex)

 A cucurbitur [girryl-D-galactosamine hydrochloride complex was prepared in the same manner as in Example 1 except that D-galactosamine hydrochloride was used instead of D-glucosamine hydrochloride in Example 1.

 [115] D-galactosamine hydrochloride (2) is a C4-digit ΟΗ

The functional group is a C4 epimer of 1 present in the vertical direction. In addition, due to variable rotation at D ₂ 0, alpha and beta anomers are present at a ratio of 63:37 after 24 hours. The high field shift of the signal in the N-NMR spectrum (298 K) results in divalent CB [ It can be seen that the tile complex was formed, which can be confirmed by MALDI-TOF (FIG. 7 and FIG. 8). For 2, the NMR spectrum is clearer than 1, and like 1, a better resolved signal appears when 2 equivalents of CB [71] are present. Favors a cyano machine there are CB [showed that the supported only odd one anomeric, proton-coupled ^l3 C-NMR a doublet _{(88.8 ppm, '7 cl.} H i = 170.7Hz) that appears in the CB ] Suggests that the two co-located alpha alphaomers are present (Fig. 9). In addition, the DQF-COSY is located in the horizontal direction.

 We can get the coupling constant (ν ^ Η ^^ πίζΛ / Η ^ Η ^ ΙΟ / ΖΗζ), which is represented by Hl (Hle), which confirms the presence of an alpha anomer (Fig. 10). No cross signal other than the correlation of H2a is shown (Fig. U).

 Comprehensive NMR analysis confirmed the presence of divalent alpha anomers during CB [7].

 [116] Methylene protons of the CB [groups generally due to the addition of guest molecules (methylene

 protons do not occur, but cyclic methylene at the carbonyl inlet

Protons (endocyclic methylene protons) are present in close proximity to guest molecules and can be identified through the ROESY spectrum through the spatial correlation between them. In the ROESY spectrum (Fig. 11) we can see that there is a spatial correlation between CB [Hie and H6 in tile 2 (5.4ppm), and H3 and H5 with large high field shifts of lOppm are Tell us that there is a deep common 2)-

[1 17] Table 2

 [Table 2]

 [1 18] This indicates that C2 and C6 of the hexameric ring during bivalent CB [

This implies that the hypotonic traversal is centered, i.e., the -NH ₃ ⁺ (C2) and -OH (Cl) functional groups of 2 point toward the carbonyl inlet of the CB group and the OH (C6) half It means that it is located in the opposite direction. The arrangement of 2 alpha anomers in CB [7] is consistent with the results of molecular mechanics modeling, indicating that Hie and H6 of 2 are in close proximity with the quantum cyclic methylene protons in the molecular model (~ 3.9 to 4.1 A). In addition, the oxygen atom of the ring is in the form of CB [71], stabilizing and ammonium ion through hydrogen bonding of the hydroxyl group of C 1 and C 2 of 2 and Stabilization through interaction of CB [7] carbonyl groups.

Due to the wideness of the signal and the rotation of the photochromic phenomenon, it is difficult to measure the binding force between CBm and 2 through the titration of ΝΜΙ (Fig. 1 3). Through ITC analysis (Table 1, Fig. 14), CB [Fig. It can be seen that it shows strong binding force ( _a = (1 .6 ± 0.1) x l0 ⁴ M- ⁱ ) and this binding force is also the value of no sugar acceptor, which is a saccharide acting in any previous water. Similarly, enthalpy and entropy change were performed in the preferred direction (Aff "=-13.8 kJ mol-', TAS" = 10.1 kJ mol ¹ ). While the enthalpy change was almost similar, 2 showed a high entropy change. The release of the hydration shell, which occurs during the formation of the complex, occurs more strongly at 2, which is considered to be a factor that enables strong bonding.

 [120] Principal guest interactions based on ion-dipole interactions of carbonyl inlets of CBm and ammonium of aminosaccharides result in relatively large negative enthalpy changes and hydrophobic interactions that are seen as positive entropy changes. It also appears to play an important role. CB [There are water molecules with high energy in the cavity of the group, which are released through the combination of guest molecules (1, 2). Although water-mediated linkage of sugars and lectins is found in many biological conditions, there have been no reports of stabilization of monosaccharide alpha anomers during the hydrophobic success of water-soluble supramolecular molecules.

 [121]

[Example 3] Cookerbitu [D-mannosamine hydrochloride (3) Manufacture of Complex (CB [7] .Triplex)

 A cucurbitur [reel -D-galactosamine hydrochloride complex was prepared in the same manner as in Example 1 except that D-mannosamine hydrochloride was used instead of D-glucosamine hydrochloride in Example 1.

 [124] D-mannosamine with ammonium ions vertically oriented at position C2

 Study on the combination of hydrochloride (D-mannosamine hydrochloride, 3) and CB [

Proceeded. 3 also shows a variable rotation at D ₂ 0 and the ratio between alpha and beta

From 24:29 it changed to 38:62 after 24 hours. CB [Title 3: 2: 1, Complex ¹

In the H-NMR spectrum (FIG. 15), it was confirmed that the signal of 3 has a high field shift due to the formation of a complex. However, unlike 1 and 2, the spectrum is complicated by the presence of two anomers. It can be seen that the americ ratio was changed in the direction of favoring alpha anomer (α: β = 65:35) due to the formation of the complex. The ITC experiment confirmed that the formation of the complex was achieved through the change of enthalpy and entropy (/ς=(1.9 ± 0.3) χ Κ Μ- ¹ ) (AH ^U = -13.0 kJ mol- ¹ ΓΔ5 "= 5.7 kJ mol of N values obtained by using the guest combination model is -0.75 (Table 1, Fig. 16) in which 3 for CB [pore perpendicular of -ΝΗ ₃ ⁺ loin present in C2 position - 1 own: ^1), 1. It seems that the orientation is affected, and the ring oxygen is exposed to the surrounding water and the rotation of the light is possible.

 [125]

 [Example 4]

 [127] Preparation of CB I * 4 complex, Cookerbitu 71 Lim-D-6-amino-6-deoxy-D-glucose.

 [128] In Example 1, instead of D-glucosamine hydrochloride, D-6-amino-6-dioxy-D-gluco

 In the same manner as in Example 1 except that it was used

 Cookerbitu [Gril-D-galactosamine hydrochloride complex was prepared.

[129] In the Ή-NMR spectrum (Fig. 17) of the same equivalent CB] and 4 complexes, it can be seen that the signal of the guest molecule 4 has high field shift, and a clearer signal can be seen when 2 equivalents of CB] are present. In-depth NMR analysis confirmed the presence of an alpha anomer in the CB [7] cavity. Also, ITC experiment showed strong binding to CB [4] (/ς=(1.6 ± 0.7) χ 10 ³ Μ- ', Table 1, Figure 18).

 [130]

 [Example] [Example 5] Cookerbitu [Gillyl-neomycin and cookerbitu [Gillyl-ferromycin complex preparation

 In Example 1 neomycin or ferromycin instead of D-glucosamine hydrochloride

 Except for the use, the same method as in Example 1 was used to prepare cookerbitu [reel-neomycin and cookerbitu [giryl-ferromycin complex].

 [133]

[134] CB [neomycin) and ITC titration as shown in Table 3 below. It can be seen that it binds strongly to paramomycin. TABLE 3

 [Table 3]

 As shown in Examples 1 to 4 of the present invention, it can be seen that the cucurbituril of the cucurbituril-sugar complex recognizes and stabilizes the sugar, and based on this, the cucurbituril can form a complex with an aminosugar antibiotic based on this. I found out.

 [137] The cooker bituril of the present invention, a sugar complex, selectively binds sugars, especially amino sugars, in an aqueous solution to strongly support amino sugars in the cooker bituril joint and prevents the rotation of amino sugars. ) To stabilize.

 [138] Also, as shown in Example 5, cooker bituriline mycin or

 It can be seen that it recognizes and supports amino sugar antibiotics such as momycin, and thus the cucurbituril-sugar complex of the present invention can be usefully used for the transport and detection of antibody.

Claims

Claims [Claim 1] Cucurbituril and, A cucurbituril-sugar complex comprising a sugar supported in a cavity of the cucurbituril. [Claim 2] In paragraph 1, The cucurbituril is a cucurbituril-sugar complex represented by Chemical Formula 1.

[In Formula 1 above, X is 0, S or NH, A, and A ₂ are each independently H, OR, SR, NHR, COOH, 0(CH ₂ ) _a R or 0(CH ₂ ) _a SR, where a is an integer from 1 to 5, and R is H , halogen, C 1 -C30 alkyl, C2-C30 alkenyl, C2-C30 alkynyl, C2-C30carbonylalkyl, C3-C30cycloalkyl, C2-C30 haterocycloalkyl, C6-C30 aryl, C6-C30 aryl, C1-C30 alkyl, C2-C30 heteroaryl or C2-C30 heteroaryl C1-C30 alkyl; n is an integer between 4 and 14.] [Claim 3] In clause 2, In Formula 1, X is 0, A, and A ₂ are each independently H, OR, 0(CH ₂ )aR or 0(CH ₂ ) aSR, where a is an integer of 1 to 5, and R is H, C1-C30 alkyl or C2-C30 alkenyl; n is an integer of 6 to 10 cucurbituril - sugar complex. [Claim 4] In paragraph 1, The sugar is a cucurbituril-saccharide complex having an amino group or a salt containing it. [Claim 5] In clause 4, The sugar is a cucurbituril-saccharide complex that is a monosaccharide, disaccharide, or polysaccharide. [Claim 6] In paragraph 4, The sugar having the above amino group or a salt containing it is Amino sugar antibiotics or those represented by the formula 2 below: Cucurbituril - sugar complex.

[In Formula 2 above, R _n to R _l5 are independently OH, NH ₂ or NH ₃ ⁺ X-, and any one of R _u to R _l5 is necessarily NH ₂ or NH ₃ ⁺ X-; X- is the monovalent anion of acid.] [Claim 7] In Article I, The cucurbituril fructose is a cucurbituril-sugar complex with an equivalence ratio of 0.5 to 3: 1. [Claim 8] Cucurbituryl-sugar is produced by contacting cucurbituryl and sugar in an aqueous solution. A method for producing a cucurbituril-saccharide complex comprising: preparing the complex. [Claim 9] In clause 8, The cucurbituril is a method for producing a cucurbituril-sugar complex represented by Chemical Formula 1.

[In Formula 1 above, X is 0, S or NH, A, and 八₂ are each independently H, OR, SR, NHR, COOH, 0(CH ₂ )a or 0(CH ₂ )aSR, where a is an integer from 1 to 5, and R is H. Halogen , C1-C30 alkyl, C2-C30 alkenyl, C2-C30 alkynyl, C2-C30carbonylalkyl, C3-C30cycloalkyl, C2-C30heterocycloalkyl, C6-C30aryl, C6-C30aryl, C1-C30alkyl C2-C30 heteroaryl or C2-C30 heteroaryl C1-C30 alkyl; n is an integer between 4 and 14.] [Claim 10] In clause 9, In Formula 1, X is o, Aᅳ and A ₂ are each independently H, OR, 0(CH ₂ )aR or 0(CH ₂ )aSR, where a is an integer of 1 to 5, and R is H, C1-C30 alkyl or C2-C30 alkenyl; n is an integer of 6 to 10 cucurbituril - method for producing a sugar complex. [Claim 11] In paragraph 8, The sugar is a cucurbituril-saccharide complex having an amino group or a salt containing it. [Claim 12] In paragraph 11, The sugar having the above amino group or a salt containing it is Method for producing an amino antibiotic or a cucurbituril-sugar complex represented by the following formula (2).

[In Formula 2 above, Ru _to R _l5 are independently OH, NH ₂ or NH ₃ ⁺ X-, and one of Ru to R ₁₅ is necessarily NH ₂ or NH ₃ ⁺ X-; X- is the 1. false anion of acid.] [Claim 13] In paragraph 8, The cucurbituril fructose is a method for producing a cucurbituril-sugar complex having an equivalence ratio of 0.5 to 3:1. ^' [Claim 14] Method for detecting sugar using cucurbituril.