Y-type biobased glycolipids, methods of making and using the same

Abstract

The application discloses Y-type bio-based glycolipid and a preparation method and application thereof, and the preparation method is as follows: taking strophanthin and beta-D-glucose pentaacetate as raw materials, under the action of a catalyst and molecular sieves in an organic solvent, a preliminary product is obtained through reaction, and then after washing, drying and removal of the organic solvent in vacuum, purification is carried out to obtain the Y-type bio-based glycolipid. The application first takes strophanthin and beta-D-glucose pentaacetate as raw materials, and through etherification reaction, a pure bio-based amphiphilic hydrophilic head group glycolipid with Y-type structure is prepared, the compactness of molecular stacking is effectively improved, the nano aggregate obtained through self-assembly has excellent thermal stability and solvent stability, the problem that a self-assembly drug carrier is applied to a biomolecular engineering field and has poor stability can be solved, and the practical application of the self-assembly material can be promoted.

Description

Technical Field

[0001] This invention relates to the field of organic chemical synthesis technology, and in particular to a Y-type bio-based glycolipid, its preparation method, and its application. Background Technology

[0002] Amphiphilic molecules can self-assemble into a series of nanomaterials with different structures, such as nanowires, nanotubes, vesicles, and hydrogels. These materials have wide applications in biomedicine, sensors, and nanocomposites, and are an important part of modern nanotechnology. The self-assembly of amphiphilic molecules is driven by non-covalent interactions such as hydrogen bonds, π-π stacking, solubilization effects, van der Waals forces, and charge transfer interactions. It is highly sensitive to molecular chemical structure, and by controlling the structure of the hydrophilic and hydrophobic parts, drastically different self-assembled nanomaterials can be obtained.

[0003] Amphiphilic molecules can be classified into petroleum-based and bio-based types according to their origin. Petroleum-based amphiphilic molecules generally suffer from poor biocompatibility, poor degradation, and high toxicity, which significantly limits their application in the biomedical field. Bio-based amphiphilic molecules, such as phospholipids, ribonucleotides, and amino acids, have attracted widespread attention in the biomedical field due to their high biocompatibility, low toxicity, and degradability. However, most of these biomolecules suffer from poor stability, making it difficult to maintain good structural stability in solvents and under varying temperatures.

[0004] Cashew nut shell oil, a plant-based renewable resource, can be processed to separate cashew phenols and cardiotonic phenols. Cashew phenols, after modification, can be used to produce a series of environmentally friendly surfactants, which have been widely applied in various fields such as oil extraction, pharmaceuticals, and cleaning agents. Current industry research reports indicate that cashew phenols can be modified to self-assemble into nanomaterials.

[0005] However, in the process of implementing the inventive technical solution in the embodiments of this application, the inventors of this application discovered that the above-mentioned technology has at least the following technical problems:

[0006] While cashew glycolipid molecules can self-assemble into nanomaterials, their stability is insufficient, particularly in terms of thermal and solvent stability, which still cannot meet the stability requirements of practical applications. Therefore, further improving the stability of bio-based self-assembled systems is of great significance for promoting their practical applications. Summary of the Invention

[0007] This invention provides a Y-type bio-based glycolipid, its preparation method, and its application, which can overcome the above-mentioned shortcomings of amphiphilic molecules in the prior art.

[0008] To address the aforementioned technical problems, this invention provides a Y-type bio-based glycolipid, comprising compounds with the following structural formula:

[0009]

[0010] Where R is C 15 H 31-2n n = 0 - 3, and

[0011] When n=0, C 15 H 31 for

[0012] When n=1, C 15 H 29 for

[0013] When n=2, C 15 H 27 for

[0014] When n=3, C 15 H 25 for

[0015] To address the aforementioned technical problems, this invention also provides a method for preparing Y-type bio-based glycolipids, comprising the following steps:

[0016] Using cardiac glycosides and β-D-glucose pentaacetate as raw materials, a preliminary product was obtained by reacting in an organic solvent under the action of a catalyst and molecular sieve. After washing, drying, and vacuum removal of the organic solvent, the product was purified to obtain the Y-type bio-based glycolipid.

[0017] In a preferred embodiment of the present invention, the molar ratio of cardiac glycoside and β-D-glucose pentaacetate is 1:2 to 8.

[0018] In a preferred embodiment of the present invention, the molar ratio of cardiac glycogen to catalyst is 1:2 to 4.

[0019] In a preferred embodiment of the present invention, the reaction temperature is 25–40°C and the reaction time is 6–36 h.

[0020] To address the aforementioned technical problems, the present invention also provides an aggregate of Y-type bio-based glycolipids, which is self-assembled from the aforementioned Y-type bio-based glycolipids.

[0021] In a preferred embodiment of the present invention, the size of the aggregate is in the nanometer range.

[0022] In a preferred embodiment of the present invention, the self-assembly step of the aggregate is as follows: the Y-type bio-based glycolipid is boiled in deionized water to obtain a clear solution, and then naturally cooled to room temperature and left to stand for a certain period of time.

[0023] In a preferred embodiment of the present invention, the boiling time is 1 to 5 hours, and the settling time is 1 to 7 days.

[0024] To address the aforementioned technical problems, the present invention also provides an application of Y-type bio-based glycolipids or aggregates thereof as drug carriers.

[0025] The beneficial effects of this invention are as follows: This invention provides a Y-type bio-based glycolipid, its preparation method, and its application. For the first time, a bio-based amphiphilic head glycolipid with a Y-type structure is prepared by using cardiac glycosides and β-D-glucose pentaacetate as raw materials through an etherification reaction. This effectively improves the compactness of intermolecular stacking. The nano-aggregates obtained by its self-assembly have excellent thermal and solvent stability and can be used as a drug carrier with excellent stability in the field of biomolecular engineering, effectively promoting the practical application of self-assembled materials. Attached Figure Description

[0026] Figure 1 This is a liquid chromatogram of the mixture of Y-type bio-based glycolipid, cardiac glycoside, and β-D-glucose pentaacetate prepared in Example 1 of this invention;

[0027] Figure 2 The Y-type bio-based glycolipid and cardiac glycoside prepared in Example 1 of this invention 1 HNMR test pattern;

[0028] Figure 3 This is an FTIR chromatogram of the Y-type bio-based glycolipid, cardiac glycoside, and β-D-glucose pentaacetate mixture prepared in Example 1 of this invention;

[0029] Figure 4 This is an LCMS test image of the Y-type bio-based glycolipid prepared in Example 1 of the present invention;

[0030] Figure 5 These are TEM images of monohead glycolipids and Y-type bio-based glycolipid aggregates prepared in Comparative Examples 1, 2 and 3 after 7 days of aging.

[0031] Figure 6 These are TEM images of the aggregates of monohead glycolipids and Y-type bio-based glycolipids prepared in Comparative Example 2 and Example 3 after aging for 30 days.

[0032] Figure 7 The images show SEM images of the aggregates of monohead glycolipids and Y-type bio-based glycolipids prepared in Comparative Example 2 and Example 3 after aging for 7 days and drying at 150°C for 20 min.

[0033] Figure 8 The images show SEM images and corresponding Tyndall effect images of the aggregates of mono-head glycolipids and Y-type bio-based glycolipids prepared in Comparative Example 2 and Example 3 after aging for 7 days and immersion in 0.9% saline for 7 days.

[0034] Figure 9 The images show SEM images of the aggregates of monohead glycolipids and Y-type bio-based glycolipids prepared in Comparative Example 2 and Example 3 after aging for 7 days and immersion in 45 wt% ethanol for 60 days. Detailed Implementation

[0035] The preferred embodiments of the present invention will now be described in detail with reference to the accompanying drawings, so that the advantages and features of the present invention can be more easily understood by those skilled in the art, thereby providing a clearer and more explicit definition of the scope of protection of the present invention.

[0036] For self-assembled materials, since molecules are bonded together through non-covalent interactions such as hydrogen bonds and hydrophobic interactions, the stability of self-assembled materials to heat and solvents is usually poor. This is one of the key technical challenges that limits the practical application of such self-assembled materials.

[0037] This invention, for the first time, uses cardiac glycolipids with symmetrical bisphenol hydroxyl groups as a raw material and synthesizes a novel bio-based amphiphilic molecule with a Y-shaped structure through reaction with β-D-glucose pentaacetate. Due to the presence of the Y-shaped hydrophilic head groups, this amphiphilic molecule exhibits a more compact stacking structure during self-assembly, which enhances the structural stability of the aggregates. Furthermore, the multiple -OH groups within the hydrophilic head groups can form hydrogen bonds, further strengthening the overall hydrogen bonding of the molecule and contributing to its structural stability. The nanoscale aggregates obtained by the self-assembly of the Y-shaped bio-based glycolipid of this invention exhibit excellent thermal and solvent stability.

[0038] Specifically, the structural formula of the Y-type bio-based glycolipid of the present invention is as follows:

[0039]

[0040] Where R is C 15 H 31-2n n = 0 - 3, and

[0041] When n=0, C 15 H 31 for

[0042] When n=1, C 15 H 29 for

[0043] When n=2, C 15 H 27 for

[0044] When n=3, C 15 H 25 for

[0045] The preparation method and principle of the Y-type bio-based glycolipid with the above structure are as follows:

[0046] (1) Under an inert gas environment, cardiac glycoside, β-D-glucose pentaacetate and boron trifluoride ether catalyst are dissolved in an organic solvent such as dichloromethane in a certain proportion, and then 4A molecular sieve is added. The mixture is stirred at 25-40℃ for 6-36 hours to obtain preliminary product A.

[0047] The cardiac glycoside, β-D-glucose pentaacetate, catalyst, and 4A molecular sieve are all pre-dried to remove water. Specifically, the cardiac glycoside, β-D-glucose pentaacetate, and catalyst are dried in an oven at 30°C for 1 hour. The 4A molecular sieve is dried in a muffle furnace at 350°C under normal pressure for 8 hours to enable it to adsorb.

[0048] The molar ratio of cardiac glycoside and β-D-glucose pentaacetate is 1:2 to 8.

[0049] The molar ratio of cardiac glycoside and the catalyst boron trifluoride ether is 1:2 to 4.

[0050] (2) The above preliminary product A was washed once with a 5% NaHCO3 solution and once with deionized water, then dried with anhydrous Na2SO4, and finally the organic solvent was removed for 1-3 hours at a temperature of 25-40℃ and a vacuum of 0-0.005MPa to obtain crude product B.

[0051] In the above reaction process, β-D-glucose pentaacetate first undergoes deethylation under acidic conditions provided by the phenolic hydroxyl group, and then generates acetic acid and intermediate product M under the action of boron trifluoride diethyl ether catalyst. In the subsequent washing step, NaHCO3 solution is used for washing, which neutralizes the acetic acid and simultaneously hydrolyzes the ester group in M ​​to generate β-cainol (taking complete hydrolysis as an example), as shown in the following reaction formula:

[0052]

[0053] (3) Add the boiled ethanol to the crude product B and continue boiling for 1 to 5 hours. Then let it cool naturally and crystallize and purify in ethanol to obtain the target product Y-type bio-based glycolipid C.

[0054] Note: The cardiac phenols used in the following examples are all from Changshu Naisu Biomaterials Technology Co., Ltd., product number 1203A, and are obtained through cashew nut shell oil distillation. The relative contents of mono-, di-, and trienes in cardiac phenol 1203A were determined by liquid chromatography, and its average relative molecular mass was calculated to be 316.

[0055] Example 1

[0056] First, the raw materials, cardiotonic phenol, β-D-glucose pentaacetate, and the catalyst boron trifluoride ether, were dried in an oven at 30°C for 1 hour. The 4A molecular sieve was then dried in a muffle furnace at 350°C under normal pressure for 8 hours to enable it to adsorb.

[0057] Cardiac glycosides, β-D-glucose pentaacetate, and boron trifluoride diethyl ether in a molar ratio of 1:2:2 were dissolved in 40 ml of dichloromethane. 2 g of 4A molecular sieve was added, and the mixture was stirred at 25 °C for 6 h to obtain the preliminary product.

[0058] The preliminary product was washed once with 10 ml of 5% sodium bicarbonate solution and once with 10 ml of deionized water. Then, 5 g of anhydrous Na₂SO₄ was added and the product was dried for 3 hours. Subsequently, dichloromethane was removed under vacuum at 25°C and 0.005 MPa for 1 hour to obtain the crude product. 100 ml of boiling ethanol was added to the crude product and boiling was continued for 1 hour. The product was then allowed to cool naturally and crystallize. The resulting precipitate was the purified target product, type Y bio-based glycolipid, with a yield of 90.2%.

[0059] Preparation of aggregates: The obtained Y-type bio-based glycolipid was boiled in deionized water for 1 hour to obtain a clear solution, which was then naturally cooled to room temperature and left for 1 day to self-assemble into nano-aggregates.

[0060] Example 2

[0061] First, the raw materials, cardiotonic phenol, β-D-glucose pentaacetate, and the catalyst boron trifluoride ether, were dried in an oven at 30°C for 1 hour. The 4A molecular sieve was then dried in a muffle furnace at 350°C under normal pressure for 8 hours to enable it to adsorb.

[0062] Cardiac glycosides, β-D-glucose pentaacetate, and boron trifluoride diethyl ether in a molar ratio of 1:2:3 were dissolved in 40 ml of dichloromethane. 2 g of 4A molecular sieve was added, and the mixture was stirred at 25 °C for 24 h to obtain the preliminary product.

[0063] The preliminary product was washed once each with 10 ml of 5% sodium bicarbonate solution and deionized water, then dried with 5 g of anhydrous Na₂SO₄ for 3 h. Subsequently, it was vacuum-treated at 35 °C and 0.005 MPa for 2 h to remove dichloromethane, yielding the crude product. 100 ml of boiling ethanol was added to the crude product, and boiling continued for 5 h. The product was then allowed to cool naturally and crystallize. The resulting precipitate was the purified target product, type Y bio-based glycolipid, with a yield of 92.9%.

[0064] Preparation of aggregates: The obtained Y-type bio-based glycolipid was boiled in deionized water for 5 hours to obtain a clear solution, which was then naturally cooled to room temperature and left for 7 days to self-assemble into nano-aggregates.

[0065] Example 3

[0066] First, the raw materials, cardiotonic phenol, β-D-glucose pentaacetate, and the catalyst boron trifluoride ether, were dried in an oven at 30°C for 1 hour. The 4A molecular sieve was then dried in a muffle furnace at 350°C under normal pressure for 8 hours to enable it to adsorb.

[0067] Cardiac glycosides, β-D-glucose pentaacetate, and boron trifluoride diethyl ether in a molar ratio of 1:8:4 were dissolved in 40 ml of dichloromethane. 2 g of 4A molecular sieve was added, and the mixture was stirred at 40 °C for 36 h to obtain the preliminary product.

[0068] The preliminary product was washed once with 10 ml of 5% sodium bicarbonate solution and once with 10 ml of deionized water. Then, 5 g of anhydrous Na₂SO₄ was added and the product was dried for 3 h. Subsequently, dichloromethane was removed under vacuum at 40 °C and 0.005 MPa for 3 h to obtain the crude product. 100 ml of boiling ethanol was added to the crude product and boiling was continued for 5 h. The product was then allowed to cool naturally and crystallize. The precipitate obtained was the purified target product, type Y bio-based glycolipid, with a yield of 94.6%.

[0069] Preparation of aggregates: The obtained Y-type bio-based glycolipid was boiled in deionized water for 5 hours to obtain a clear solution, which was then naturally cooled to room temperature and left for 7 days to self-assemble into nano-aggregates.

[0070] Example 4

[0071] First, the raw materials, cardiotonic phenol, β-D-glucose pentaacetate and the catalyst boron trifluoride ether, were dried in an oven at 30°C for 1 hour. Then, the 4A molecular sieve was dried in a muffle furnace at 350°C under normal pressure for 8 hours to enable it to adsorb.

[0072] Cardiac glycosides, β-D-glucose pentaacetate, and boron trifluoride diethyl ether in a molar ratio of 1:2:2 were dissolved in 40 ml of dichloromethane. 2 g of 4A molecular sieve was added, and the mixture was stirred at 25 °C for 30 h to obtain the preliminary product.

[0073] The preliminary product was washed once each with 10 ml of 5% sodium bicarbonate solution and 10 ml of deionized water, then dried with 5 g of anhydrous Na₂SO₄ for 3 h. Subsequently, it was vacuum-treated at 25 °C for 2 h to remove dichloromethane, yielding the crude product. 100 ml of boiling ethanol was added to the crude product, and boiling continued for 2 h. The product was then allowed to cool naturally and crystallize. The resulting precipitate was the purified target product, type Y bio-based glycolipid, with a yield of 90.9%.

[0074] Preparation of aggregates: The obtained Y-type bio-based glycolipid was boiled in deionized water for 2 hours to obtain a clear solution, which was then naturally cooled to room temperature and left for 2 days to self-assemble into nano-aggregates.

[0075] Example 5

[0076] First, the raw materials, cardiotonic phenol, β-D-glucose pentaacetate, and the catalyst boron trifluoride ether, were dried in an oven at 30°C for 1 hour. The 4A molecular sieve was then dried in a muffle furnace at 350°C under normal pressure for 8 hours to enable it to adsorb.

[0077] Cardiac glycosides, β-D-glucose pentaacetate, and boron trifluoride diethyl ether in a molar ratio of 1:5:3 were dissolved in 40 ml of dichloromethane. 2 g of 4A molecular sieve was added, and the mixture was stirred at 35 °C for 21 h to obtain the preliminary product.

[0078] The preliminary product was washed once with 10 ml of 5% sodium bicarbonate solution and once with 10 ml of deionized water. Then, 5 g of anhydrous Na₂SO₄ was added and the product was dried for 3 hours. Subsequently, dichloromethane was removed under vacuum at 35°C and 0.005 MPa for 2 hours to obtain the crude product. 100 ml of boiling ethanol was added to the crude product and boiling was continued for 3 hours. The product was then allowed to cool naturally and crystallize. The precipitate obtained was the purified target product, type Y bio-based glycolipid, with a yield of 92.1%.

[0079] Preparation of aggregates: The obtained Y-type bio-based glycolipid was boiled in deionized water for 3 hours to obtain a clear solution, which was then naturally cooled to room temperature and left for 5 days to self-assemble into nano-aggregates.

[0080] Comparative Example 1

[0081] Pentadecylphenol, β-D-glucose pentaacetate, boron trifluoride ether catalyst, and 4A molecular sieve were dried using the drying methods described in Examples 1-5.

[0082] Pentadecylphenol, β-D-glucose pentaacetate, and boron trifluoride diethyl ether in a molar ratio of 1:4:2 were dissolved in 40 ml of dichloromethane. 2 g of 4A molecular sieve was added, and the mixture was stirred at 40 °C for 36 h to obtain the preliminary product.

[0083] The preliminary product was washed once with 10 ml of 5% sodium bicarbonate solution and once with 10 ml of deionized water. Then, 5 g of anhydrous Na₂SO₄ was added and the product was dried for 3 hours. Subsequently, dichloromethane was removed under vacuum at 40°C and 0.005 MPa for 3 hours to obtain the crude product. 100 ml of boiling ethanol was added to the crude product and boiling was continued for 5 hours. The product was then allowed to cool naturally and crystallize. The resulting precipitate was the purified target product, glycolipid.

[0084] Preparation of aggregates: The obtained glycolipid was boiled in deionized water for 5 hours to obtain a clear solution, which was then naturally cooled to room temperature and left for 7 days to self-assemble into nano-aggregates.

[0085] Comparative Example 2

[0086] Cashew phenol, β-D-glucose pentaacetate, boron trifluoride ether catalyst, and 4A molecular sieve were dried using the drying methods described in Examples 1-5.

[0087] Cashew phenol, β-D-glucose pentaacetate and boron trifluoride diethyl ether in a molar ratio of 1:4:2 were dissolved in 40 ml of dichloromethane, and 2 g of 4A molecular sieve was added. The mixture was stirred at 40 °C for 36 h to obtain the preliminary product.

[0088] The preliminary product was washed once with 10 ml of 5% sodium bicarbonate solution and once with 10 ml of deionized water. Then, 5 g of anhydrous Na₂SO₄ was added and the product was dried for 3 hours. Subsequently, dichloromethane was removed under vacuum at 40°C and 0.005 MPa for 3 hours to obtain the crude product. 100 ml of boiling ethanol was added to the crude product and boiling was continued for 5 hours. The product was then allowed to cool naturally and crystallize. The resulting precipitate was the purified target product, a glycolipid.

[0089] Preparation of aggregates: The obtained glycolipid was boiled in deionized water for 5 hours to obtain a clear solution, which was then naturally cooled to room temperature and left for 7 days to self-assemble into nano-aggregates.

[0090] The morphology and stability of the glycolipids prepared in Examples 1-5 and Comparative Examples 1-2, and the nanoaggregates obtained by their self-assembly, were tested below.

[0091] (I) Verification of successful synthesis of Y-type bio-based glycolipid

[0092] 1. Liquid Chromatography Test

[0093] The raw materials, cardiotonic phenol and β-D-glucose pentaacetate, were mixed in a 1:1 molar ratio, and then dichloromethane solvent was added and stirred for 24 hours. The solvent was removed by rotary evaporation and used as a control sample.

[0094] The molecular structure of the Y-type bio-based glycolipid prepared in Example 1 was characterized by liquid chromatography, and the test results are shown in the figure below. Figure 1 As shown.

[0095] The liquid chromatography conditions are as follows:

[0096] Standard substance: methanol;

[0097] Column: CNWAthena C18, column temperature 35℃;

[0098] Mobile phase: methanol;

[0099] Mobile phase gradient: 1 ml / min;

[0100] Tested ultraviolet light wavelength: 273nm.

[0101] The control sample prepared above was characterized by liquid chromatography, and the test chromatogram is shown in the figure. Figure 1 As shown.

[0102] like Figure 1 As shown, peaks 1-4 are bisphenol peaks, and peaks 5-8 are monophenol peaks.

[0103] The structural formula corresponding to the bisphenol peak is:

[0104] Figure 1 In the peak corresponding to 1,

[0105] In the peak corresponding to 2,

[0106] In the peak corresponding to 3,

[0107] In the peak corresponding to 4,

[0108] The structural formula corresponding to the monophenol peak is:

[0109] Figure 1 In the middle, among the peaks corresponding to 5,

[0110] In the peak corresponding to 6,

[0111] In the peak corresponding to 7,

[0112] In the peak corresponding to 8,

[0113] In the HPLC chromatogram of the control sample, no new peaks different from those of the raw materials appeared in the peak positions of cardiac glycophenol and β-D-glucose pentaacetate. However, in the HPLC chromatogram of glycolipid molecules, the peak intensities of the raw material peaks were all reduced, and glycolipid peaks different from those of the two raw materials appeared in the 2-3 min range.

[0114] 2. NMR analysis

[0115] The synthesis of a type Y glycolipid from the reaction of cardiac glycosides and β-D-glucose pentaacetate occurs at the phenolic hydroxyl group of the cardiac glycosides, as confirmed by NMR spectroscopy. 1 The structures of the raw material cardiac glycoside and the prepared Y-type glycolipid were characterized by HNMR, and the test results are shown in the figure below. Figure 2 As shown.

[0116] Depend on Figure 2 It can be known that cardiac glycosides... 1 In the HNMR spectrum, the peak at 5.71 ppm corresponds to the proton in the phenolic hydroxyl group, i.e., position 1 in the spectrum. Meanwhile, the glycolipid molecule... 1 The disappearance of this peak in the HNMR spectrum indicates that the synthesis of Y-type glycolipids occurs at the phenolic hydroxyl group of cardiac glycolipids.

[0117] 3. FT-IR test

[0118] To further prove that the above-mentioned Y-type glycolipid is formed by the dehydration condensation of the phenolic hydroxyl group of cardiac glycoside and the hemiacetal hydroxyl group generated by the hydrolysis of β-D-glucose pentaacetate, its structure was further analyzed by FTIR based on liquid chromatography.

[0119] Specifically, the molecular structure of the Y-type bio-based glycolipid prepared in Example 1 was characterized using FT-IR, and the test results are shown in the figure below. Figure 3 As shown.

[0120] The structures of the raw materials cardiac glycoside and β-D-glucose pentaacetate were characterized by FT-IR, respectively. As a control, the test results are shown in the figure below. Figure 3 As shown.

[0121] Depend on Figure 3 It can be seen that in the FTIR spectrum of β-D-glucose pentaacetate, 1042 cm⁻¹ -1 and 1070cm -1 The peak at 2924 cm⁻¹ is the saturated cyclic COC peak in glucose. In the cardiac phenol spectrum, 2924 cm⁻¹ is the peak at 2924 cm⁻¹. -1 and 2853cm -1 These peaks represent the stretching vibrations of -CH2 and -CH3, respectively. All of these peaks were also observed in the FTIR spectra of type Y glycolipids. In addition, the 1229 cm⁻¹ peak in type Y glycolipids... -1The new peak at that point represents the aryl alkyl ether COC peak. This demonstrates that the dehydration condensation reaction in the synthesis of type Y glycolipids occurs at the phenolic hydroxyl group of the cardiac glycoside and the hemiacetal hydroxyl group of glucose.

[0122] 4. LCMS Analysis

[0123] To further demonstrate that both phenolic hydroxyl groups of the cardiac glycolipin participated in the reaction to form two glycosidic bonds, the intermediate product M of the Y-type bio-based glycolipid synthesized in Example 1 was subjected to LCMS analysis. The test results are as follows: Figure 4 As shown.

[0124] The test conditions are as follows:

[0125] Column: Agilent Technologies, Inc. EclipsePlus-C18, 2.1 × 500 mm, 1.8 μm;

[0126] Column temperature: 25℃;

[0127] Mobile phase: A. Water, B. Methanol (5:95, V:V);

[0128] Flow rate: 0.3 mL / min;

[0129] Injection volume: 1 μL.

[0130] Mass spectrometry acquisition parameters:

[0131] Ion source type: ESI;

[0132] Mode: MS2 Scan;

[0133] Polarity: negative;

[0134] Range: 100-1000 m / z;

[0135] Ion source temperature: 350℃;

[0136] Atomizing gas flow rate and pressure: 10 L / min, 45 psi;

[0137] Capillary voltage: 3500V; Fragmentor: 100V;

[0138] Scan step size: 0.1 amu.

[0139] Mass spectrum extraction method: average mass spectrum of the 10% peak height portion.

[0140] Since the fully hydrolyzed glycosyl-modified cashew phenol molecules are insoluble in organic solvents such as dichloromethane, we chose to characterize the unhydrolyzed intermediate product M by LC-MS. Comparing the mass spectrometric responses of the glycolipid in positive and negative ion modes revealed a very low response in negative ion mode, while the response reached 10⁵ in positive ion mode. Therefore, we selected the secondary mass spectrum in positive ion mode for fragment ion analysis, such as... Figure 4 As shown. Among them. Figure 4 Figures a-4c show the secondary mass spectra of the synthesized glycolipid molecules, while figures a1-c2 show the specific glycolipid molecule structures and their breakage locations corresponding to each mass spectrum.

[0141] Figure 4 The fragment ion peaks at m / z 330.7 and m / z 646.8 appearing in the secondary mass spectrum of the quasi-molecular ion peak [M+H]+ at m / z 977.3, are from the quasi-molecular ion. Figure 4 Fragments resulting from a fracture at the location shown in Figure a1. Similarly, Figure 4 The fragment ion peaks at m / z 330.8 and m / z 647.8 appearing in the secondary mass spectrum of the quasi-molecular ion peak [M+H]+ at m / z 979.5 shown in b are from... Figure 4 It was caused by a fracture at the location shown in b1. For Figure 4 c. In the secondary mass spectrum of the quasi-molecular ion peak [M+H]+ at m / z 981.6, a fragment ion peak at m / z 674.8 appeared. Analysis indicates that the quasi-molecular ion originated from... Figure 4 Fragments formed by breakage at position c1. The fragment ion peak at m / z 369.9 represents quasi-ionic molecules in... Figure 4 Ionic fragments generated by the fracture at position 1 shown in c2; based on this fracture, the alkyl side chain R=C 15 H 31 Fragmentation occurred again at positions 2 and 3, respectively, resulting in fragment ion peaks at m / z 356 and m / z 338.1.

[0142] (II) Morphology Characterization of Self-Assembled Nanomaterials

[0143] To observe the morphology of aggregates formed by the self-assembly of glycolipid molecules with different structures at different aging times, the self-assembled aggregates in Example 3, Comparative Example 1, and Comparative Example 2 were aged for 7 days and 30 days, respectively, and the morphology of the formed nanomaterial aggregates was observed by SEM and TEM.

[0144] 1. TEM morphology after 7 days of aging

[0145] Figure 5 (a) is a TEM image of the monohead glycolipid aggregate synthesized from pentadecylphenol in Comparative Example 1 after 7 days of aging.

[0146] Figure 5 (b) is a TEM image of nanofibers formed from monohead glycolipid aggregates synthesized from cashew phenol in Comparative Example 2 after 7 days of aging. Figure 5 (c) is Figure 5 Enlarged view of the boxed area in (b). Figure 5 (d) is a TEM image of nanotubes formed by the single-headed glycolipid aggregate synthesized from cashew phenol as a raw material in Comparative Example 2 after 7 days of aging.

[0147] Figure 5 Image (ef) is a TEM image of the Y-type glycolipid aggregate synthesized from cardiac glycophenol in Example 3 after 7 days of aging.

[0148] Depend on Figure 5 As can be seen from Figures 5(a) and 5(b), the fiber morphology formed by the two glycolipid aggregates is the same, with the fibers intertwined and interwoven, and the fibers are not hollow.

[0149] Depend on Figure 5 In (c), it can be observed that there is a clear orientation in all directions within the fiber. This is because pentadecylphenol has C 15 The side chains are all saturated carbon, and the self-assembly process will only form nanofiber structures; while cashew phenol's C 15 The presence of unsaturated double bonds in the side chains leads to reduced crystallinity of the nanostructures, making them more prone to self-assembly into nanotubes. Therefore, the presence of nanotubes was observed in self-assembled samples of glycolipids synthesized from cashew nutshellol, such as... Figure 5 As shown in (d). Due to the relatively short aging time, the resulting nanotubes are small in size (approximately 600 nm in length and 145 nm in width).

[0150] Depend on Figure 5 In (e), it can be observed that the Y-type glycolipid self-assembles into a stacked sheet structure. Figure 5 (f) is Figure 5 (e) Enlarged view of a certain region. The figure shows that the orientation within the lamellar structure of the Y-type glycolipid aggregates is all in the same direction. The reason for these different structures and orientations is that in an aqueous environment, the symmetrical amphiphilic head groups of the Y-type glycolipid tend to arrange themselves side-by-side, thus forming a lamellar structure, such as... Figure 5 As shown in (e). Because the long side chains in the cardiac glycoside structure only have cis structures, therefore Figure 5 The unified orientation observed in (f) is the side-by-side arrangement of C 15 Side chains. Monocapsidic glycolipid molecules lack symmetry and tend to form randomly intertwined fibers, C 15 The irregular arrangement of side chains creates different orientations within the fiber.

[0151] 2. TEM morphology after 30 days of aging

[0152] Figure 6 The image in the middle (ac) is a TEM image of the monohead glycolipid aggregate synthesized from cashew phenol as a raw material in Comparative Example 2 after 30 days of aging.

[0153] Figure 6 The image in (df) is a TEM image of the Y-type glycolipid aggregate synthesized from cardiac glycophenol in Example 3 after 30 days of aging.

[0154] Depend on Figure 6 In (a), it can be observed that after 30 days of aging, the self-assembled structure of monohead glycolipid molecules changed from nanofibers to a large number of nanotube structures, and the nanotubes were larger than those formed after 7 days of aging.

[0155] Figure 6 In the middle (d) to the middle (f) of the sixth, only tubular structures were observed. Unlike monohead glycolipids, the nanotube morphology of the aggregates formed by the self-assembly of Y-type glycolipids all showed branching.

[0156] (III) Stability Analysis of Nanoaggregates

[0157] 1. Thermal stability analysis

[0158] The monocaptopy glycolipid aggregates prepared in Comparative Example 2 were aged for 7 days, dried at 150°C for 20 min, and then characterized by SEM. The test images are shown below. Figure 7 As shown in (a).

[0159] The Y-type glycolipid aggregates prepared in Example 3 were aged for 7 days, dried at 150°C for 20 minutes, and then characterized by SEM. The test images are shown below. Figure 7 As shown in (b).

[0160] Depend on Figure 7 (a) and Figure 7 As shown in (b), helical fiber structures were observed in the self-assembled samples of mono-head glycolipids and Y-type glycolipids after drying at 150°C. The analysis suggests that this was due to the influence of the cashew phenol side chain in the glycolipids on the chirality of glucose.

[0161] but Figure 7 The fibrous structure formed by monocephalic glycolipids, as shown in (a), has been significantly disrupted and degraded; while Figure 7 The SEM image of the Y-type glycolipid aggregate shown in (b) still shows complete helical fibers and nanotubes, indicating that it can exist stably at a high temperature of 150°C, which shows that the thermal stability of the Y-type glycolipid aggregate is better than that of the monohead glycolipid aggregate.

[0162] 2. Stability analysis under physiological saline concentration conditions in the human body

[0163] The monocapsidic glycolipid aggregates prepared in Comparative Example 2 were aged for 7 days and then immersed in 0.9% saline for 7 days. SEM images and Tyndall effect diagrams were then recorded. Figure 8 As shown in (a).

[0164] The Y-type glycolipid aggregates prepared in Example 3 were aged for 7 days and then immersed in 0.9% physiological saline for 7 days. SEM images and Tyndall effect diagrams were then recorded. Figure 8 As shown in (b).

[0165] Depend on Figure 8 As shown in (a), the Tyndall effect was not observed in the clear solution obtained from self-assembly, and a large amount of flocculent material was deposited at the bottom. This is because the monohead glycolipid molecules in the aggregate structure, which were originally stable by the double-layer force (repulsion), were affected by the Na+ in the saline solution. + and Cl - The charge effect weakens the intermolecular repulsion, causing two molecules to aggregate together and form flocculent structures, indicating that the aggregate structure formed by monocapsid glycolipids cannot exist stably in 0.9% saline.

[0166] Depend on Figure 8 As shown in (b), the aggregate structure formed by Y-type glycolipids did not exhibit obvious aggregation, but maintained its cross-entangled long fiber structure, and a clear Tyndall effect was observed in the self-assembled clear solution. This indicates that the aggregate structure formed by Y-type glycolipids remained stable in physiological saline.

[0167] 3. Stability analysis in solvent (ethanol)

[0168] The monocaptopy glycolipid aggregates prepared in Comparative Example 2 were aged for 7 days and then immersed in 45 wt% ethanol for 60 days. SEM images were then recorded, as shown below. Figure 9 As shown in (a).

[0169] The bis(hydroxyglycolipid) aggregates prepared in Example 3 were aged for 7 days and then immersed in 45 wt% ethanol for 60 days. SEM images were then recorded, as shown below. Figure 9 As shown in (bd).

[0170] Depend on Figure 9 As shown in (a), no nanostructures were observed on the silicon wafer surface after the aggregate structure formed by monohead glycolipids was soaked in ethanol.

[0171] Depend on Figure 9 As shown in (b), a small number of nanotubes were observed on the silicon wafer surface after the Y-type glycolipid aggregate structure was immersed in ethanol. Figure 9 (c) and Figure 9As can be seen in (d), the size of these nanotubes is all in the hundreds of micrometers, and the tubes are not hollow in the middle. This indicates that the aggregate structure formed by Y-type glycolipids is more stable in ethanol than that formed by monohead glycolipids.

[0172] The Y-type bio-based glycolipid prepared by this invention has the following advantages:

[0173] 1. The introduction of a Y-shaped structure transforms the original linear arrangement of hydrophilic and oleophilic ends into a Y-shaped structure, resulting in a denser stacking structure.

[0174] 2. The multiple -OH groups in the disaccharides enhance hydrogen bonding, and the self-assembled nanoaggregates have higher solvent stability and thermal stability. They also have the advantages of high biocompatibility, easy degradation and low toxicity of bio-based materials, and can be used as a drug carrier with excellent stability in the field of biomolecular engineering.

[0175] The above description is merely an embodiment of the present invention and does not limit the patent scope of the present invention. Any equivalent transformations made based on the content of the present invention specification and drawings, or direct or indirect applications in other related technical fields, are similarly included within the patent protection scope of the present invention.

Claims

1. A Y-type bio-based glycolipid, characterized in that, Compounds including those with the following structural formulas: Where R is C 15 H 31-2n n = 0 - 3, and When n=0, C 15 H 31 for When n=1, C 15 H 29 for When n=2, C 15 H 27 for When n=3, C 15 H 25 for 2. A method for preparing the Y-type bio-based glycolipid as described in claim 1, characterized in that, Includes the following steps: Using cardiac glycosides and β-D-glucose pentaacetate as raw materials, a preliminary product was obtained by reacting in an organic solvent under the action of a catalyst and molecular sieve. After washing, drying, and vacuum removal of the organic solvent, the product was purified to obtain the Y-type bio-based glycolipid.

3. The method for preparing Y-type bio-based glycolipids according to claim 2, characterized in that, The molar ratio of cardiac glycoside and β-D-glucose pentaacetate is 1:2 to 8.

4. The method for preparing Y-type bio-based glycolipids according to claim 2, characterized in that, The molar ratio of cardiac glycoside to catalyst is 1:2 to 4.

5. The method for preparing Y-type bio-based glycolipids according to claim 2, characterized in that, The reaction is carried out at a temperature of 25–40°C for a duration of 6–36 hours.

6. An aggregate of Y-type bio-based glycolipids, characterized in that, It is self-assembled from the Y-type bio-based glycolipid described in claim 1.

7. The Y-type bio-based glycolipid aggregate according to claim 6, characterized in that, The aggregates are nanometer-sized.

8. The Y-type bio-based glycolipid aggregate according to claim 7, characterized in that, The self-assembly step of the aggregate is as follows: the Y-type bio-based glycolipid is boiled in deionized water to obtain a clear solution, which is then naturally cooled to room temperature and left to stand for a period of time.

9. The Y-type bio-based glycolipid aggregate according to claim 8, characterized in that, The boiling time is 1 to 5 hours, and the settling time is 1 to 7 days.

10. The application of the Y-type bio-based glycolipid of claim 1 or the aggregate of claim 7, characterized in that, Used as a drug carrier.

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