SUPRAMOLECULAR CELL-BASED CARRIER, DRUG LOADING SYSTEM AND ITS PREPARATION METHOD

Abstract

Disclosed is a method for preparing supramolecular cell-based carrier, which relates to the technical fields of supramolecular chemistry, supramolecular materials and cell preparations. Host-guest interactions mediated supramolecular cell-based carriers can achieve targeted delivery based on cell physiological functions, and have high biocompatibility, physiological barrier permeability, and targeting delivery efficiency. It does not require covalent bond modification on the cell surface, and has no effect on the physiological functions of transporting cells. The preparation method of supramolecular cell-based carrier provided by the present application has the advantages of simple and fast construction process, mild conditions and universal applicability, and the method has bio-orthogonality. In addition, a drug loading system is also provided, which can realize drug loading for targeted therapy.

Claims

1. A supramolecular cell-based carrier, comprising: a first part and a second part conjugated to each other through host-guest interaction, wherein the first part is a first cell modified by a macrocyclic host molecule or guest molecule, and the second part is a nanoparticle modified by the guest molecule or macrocyclic host molecule, or the second cell modified by the guest molecule or macrocyclic host molecule, the macrocyclic host molecule or guest molecule in the first part is embedded in the cell membrane of the first cell by coupling a membrane-embedding material, the macrocyclic host molecule or guest molecule is in correspondence with the guest molecule or macrocyclic host molecule.

2. The supramolecular cell-based carrier of claim 1, wherein the macrocyclic main molecule is cyclodextrin, cucurbituril, calixarene, pillararene or crown ether.

3. The supramolecular cell-based carrier of claim 1, wherein the first cell is selected from macrophage, neutrophil, red blood cell, stem cell, lymphocyte, dendritic cell, platelet and fat cell.

4. The supramolecular cell-based carrier of claim 1, wherein the molar ratio of the macrocyclic host molecule to the guest molecule is 1-10:1-10, the guest molecule is adamantane or ferrocene.

5. The supramolecular cell-based carrier of claim 1, wherein the nanoparticles are at least one of liposomes, micelles, nanogels, inorganic nanoparticles and nanocapsules, the second cells are liver cells, stem cells, lymphocytes, dendritic cells, platelets, fat cells or red blood cells.

6. The supramolecular cell-based carrier of claim 5, wherein the membrane-embedding material is PEG-DMPE, PEG-DPPE, PEG-DSPE or PEG-CHOL.

7. A method for preparing the supramolecular cell-based carrier of claim 1, comprising: (a) co-incubating the macrocyclic host molecule or guest molecule coupled with the membrane-embedding material with the first cell to obtain the first part; and (b) then mixing the nanoparticles modified with the guest molecules or macrocyclic host molecule, or the second cells modified with the guest molecules or macrocyclic host molecule with the first part.

8. A method for preparing the supramolecular cell-based carrier of claim 2, comprising: (a) co-incubating the macrocyclic host molecule or guest molecule coupled with the membrane-embedding material with the first cell to obtain the first part; and (b) then mixing the nanoparticles modified with the guest molecules or macrocyclic host molecule, or the second cells modified with the guest molecules or macrocyclic host molecule with the first part.

9. A method for preparing the supramolecular cell-based carrier of claim 3, comprising: (a) co-incubating the macrocyclic host molecule or guest molecule coupled with the membrane-embedding material with the first cell to obtain the first part; and (b) then mixing the nanoparticles modified with the guest molecules or macrocyclic host molecule, or the second cells modified with the guest molecules or macrocyclic host molecule with the first part.

10. A method for preparing the supramolecular cell-based carrier of claim 4, comprising: (a) co-incubating the macrocyclic host molecule or guest molecule coupled with the membrane-embedding material with the first cell to obtain the first part; and (b) then mixing the nanoparticles modified with the guest molecules or macrocyclic host molecule, or the second cells modified with the guest molecules or macrocyclic host molecule with the first part.

11. A method for preparing the supramolecular cell-based carrier of claim 5, comprising: (a) co-incubating the macrocyclic host molecule or guest molecule coupled with the membrane-embedding material with the first cell to obtain the first part; and (b) then mixing the nanoparticles modified with the guest molecules or macrocyclic host molecule, or the second cells modified with the guest molecules or macrocyclic host molecule with the first part.

12. A method for preparing the supramolecular cell-based carrier of claim 6, comprising: (a) co-incubating the macrocyclic host molecule or guest molecule coupled with the membrane-embedding material with the first cell to obtain the first part; and (b) then mixing the nanoparticles modified with the guest molecules or macrocyclic host molecule, or the second cells modified with the guest molecules or macrocyclic host molecule with the first part.

13. The preparation method of claim 7, wherein the preparation method further comprises covalently conjugating the macrocyclic molecule or guest molecule to the membrane-embedding material, and then embedding the macrocyclic molecule or guest molecule in the cell membrane of the first cell through membrane-embedding, the macrocyclic host molecule or guest molecule is covalently linked to the PEG of membrane-embedding material, the macrocyclic host molecule or guest molecule covalently conjugated with membrane-embedding material is co-incubated with the first cell for more than 30 minutes, the concentration of the macrocyclic host molecule or guest molecule covalently conjugated with membrane-embedding material is 1 μM-1 mM.

14. The preparation method of claim 7, wherein the time for mixing and incubating the nanoparticles modified with guest molecules or macrocyclic host molecule, or the second cells modified with guest molecules or macrocyclic host molecule with the first part is ≥10 seconds.

15. A drug-carrying system, comprising the supramolecular cell-based carrier of claim 1 and a drug, wherein the drug is loaded in the nanoparticle or in the second cell; the nanoparticle is a liposome.

16. A drug-carrying system, comprising the supramolecular cell-based carrier of claim 2 and a drug, wherein the drug is loaded in the nanoparticle or in the second cell; the nanoparticle is a liposome.

17. A drug-carrying system, comprising the supramolecular cell-based carrier of claim 3 and a drug, wherein the drug is loaded in the nanoparticle or in the second cell; the nanoparticle is a liposome.

18. A drug-carrying system, comprising the supramolecular cell-based carrier of claim 4 and a drug, wherein the drug is loaded in the nanoparticle or in the second cell; the nanoparticle is a liposome.

19. A drug-carrying system, comprising the supramolecular cell-based carrier of claim 5 and a drug, wherein the drug is loaded in the nanoparticle or in the second cell; the nanoparticle is a liposome.

20. A drug-carrying system, comprising the supramolecular cell-based carrier of claim 6 and a drug, wherein the drug is loaded in the nanoparticle or in the second cell; the nanoparticle is a liposome.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0046] In order to more clearly illustrate the technical solutions of the embodiments of the present disclosure, the following will briefly introduce the accompanying drawings used in the embodiments. It is to be expressly understood that the following drawings only show some examples of the present disclosure, so they should not be intended as a definition of the limits of the invention. Those skilled in this art can also obtain other related drawings based on these drawings without creative work.

[0047] FIG. 1 is a fluorescence imaging diagram of the supramolecular cell-liposome conjugate in Example 1 of the present disclosure.

[0048] FIG. 2 is a scanning electron micrograph of the supramolecular cell-liposome conjugate in Example 1 of the present disclosure.

[0049] FIG. 3 is a fluorescent imaging diagram of the supramolecular cell-cell conjugate in Example 5 of the present disclosure.

[0050] FIG. 4 is a scanning electron microscope image of the supramolecular cell-cell conjugate in Example 5 of the present disclosure.

DETAILED DESCRIPTION OF THE EMBODIMENTS

[0051] In order to make the purpose, technical solutions, and advantages of the embodiments of the present disclosure clearer, the technical solutions in the embodiments of the present disclosure will be clearly and completely described below. Those who do not indicate the specific conditions in the examples are carried out according to the conventional conditions or the conditions recommended by the manufacturer. The reagents or instruments without manufacturer information are all conventional products that could be purchased from the market.

[0052] The characteristics and performance of the present disclosure will be described in further detail below with examples.

EXAMPLE 1

[0053] This example provides a supramolecular cell-based carrier and its preparation method. In this example, the first cell is macrophage, and both DSPE-PEG-β-CD and DSPE-PEG-ADA are purchased from Xi'an ruixi Biological Technology Co., Ltd., DMEM medium was purchased from Thermo Fisher Scientific (China) Co., Ltd., and doxorubicin was purchased from Shanghai Aladdin Biochemical Technology Co., Ltd.

[0054] Macrophages were incubated in a blank DMEM medium containing 10 μM of DSPE-PEG-β-CD at 37° C. for 2 hours.

[0055] After incubation, excess DSPE-PEG-β-CD was washed away, and 10 μM of DSPE-PEG-ADA-modified doxorubicin-loaded liposomes were added to further incubate for 2 minutes.

[0056] After washing off unbound liposomes, supramolecular cell-liposome conjugates were prepared.

[0057] Since doxorubicin has red fluorescence, the prepared supramolecular cell-liposome conjugates were tracked by fluorescence imaging and scanning electron microscope imaging. The fluorescence imaging of the supramolecular cell-liposome conjugate is shown in FIG. 1, and the scanning electron microscope image of the supramolecular cell-liposome conjugate is shown in FIG. 2.

EXAMPLE 2

[0058] This example provides a supramolecular cell-based carrier and its preparation method. In this example, DSPE-PEG-β-CD and DMEM medium were purchased from the same source in Example 1. In this example, the first cell is macrophage, and the macrophages were incubated in a blank DMEM medium containing 50 μM of DSPE-PEG-β-CD at 37° C. for 1 hour.

[0059] After washing off excess DSPE-PEG-β-CD, 50 μM of DSPE-PEG-ADA modified liposomes were added and incubated for 2 minutes. Finally, after removing unbound liposomes, supramolecular cell-liposome conjugates were obtained.

EXAMPLE 3

[0060] This example provides a supramolecular cell-based carrier and its preparation method. DMPE-PEG-CB[7] (CB[7] is cucurbit[7]uril) and DMPE-PEG-ADA were prepared in laboratory. In this example, neutrophils were incubated in a blank DMEM medium containing 100 μM of DMPE-PEG-CB[7] at 37° C. for 2 hours.

[0061] After washing off excess DMPE-PEG-CB[7], 100 μM of DMPE-PEG-ADA modified liposomes were added and incubated for 1 minute. Finally, after removing unbound liposomes, supramolecular cell-liposome conjugates were obtained.

EXAMPLE 4

[0062] This example provides a supramolecular cell-based carrier and its preparation method. DPPE-PEG-CB[7] and DPPE-PEG-ADA were prepared in laboratory. In this example, hematopoietic stem cells were incubated in a blank DMEM medium containing 40 μM of DPPE-PEG-CB[7] at 37° C. for 1.5 hours.

[0063] Then excess DPPE-PEG-CB[7] was washed off, and 80 μM of DPPE-PEG-ADA modified liposomes were added to continue incubation for 5 minutes. After washing off unbound liposomes, supramolecular cell-liposome conjugates were obtained.

EXAMPLE 5

[0064] This example provides a supramolecular cell-based carrier and its preparation method. Both DiD and DiO were purchased from Shanghai Beyotime Biotechnology Co., Ltd. In this example, macrophages were incubated in a blank DMEM medium containing 10 μM of DSPE-PEG-β-CD at 37° C. for 2 hours. After washing off excess DSPE-PEG-β-CD, 10 μM of DSPE-PEG-ADA modified human hepatocytes were added and further incubated for 2 minutes. Then unbounded hepatocytes were washed off to obtain supramolecular cell-cell conjugates.

[0065] The obtained supramolecular cell-cell conjugates were studied by fluorescence imaging and scanning electron microscope imaging. The fluorescence imaging is shown in FIG. 3, and the scanning electron microscope imaging is shown in FIG. 4. Macrophages were stained with DiD (red) and human hepatocytes were stained with DiO (green).

EXAMPLE 6

[0066] This example provides a supramolecular cell-based carrier and its preparation method. Fc was purchased from Shanghai Aladdin Biochemical Technology Co., Ltd., and DSPE-PEG-Fc was synthesized in laboratory. In this example, macrophages were incubated in a blank DMEM medium containing 10 μM of DSPE-PEG-β-CD at 37° C. for 2 hours. After incubation, excess DSPE-PEG-β-CD was washed off, and 10 μM of DSPE-PEG-Fc (Fc was ferrocene) modified human hepatocytes were added to incubate for 2 minutes. Then unbound cells were washed off to obtain supramolecular cell-cell conjugates.

EXAMPLE 7

[0067] This example provides a supramolecular cell-based carrier and its preparation method. DMPE-PEG-P5 (P5 is Pillar[5]arene) was synthesized in laboratory. In this example, neutrophils were incubated at 37° C. for 2 hours in a blank DMEM medium containing 60 μM of DMPE-PEG-P5. After incubation, excess DMPE-PEG-P5 was washed off, and then 30 μM of DSPE-PEG-Fc (Fc is ferrocene) modified red blood cells were added and continued to incubate for another 2 minutes. After removing unbound cells, cell-cell conjugates were obtained.

EXAMPLE 8

[0068] This example provides a supramolecular cell-based carrier and its preparation method. Hematopoietic stem cells are derived from the American Type Culture Collection (ATCC), and both DPPE-PEG-β-CD and DPPE-PEG-ADA were prepared in laboratory. In this example, hematopoietic stem cells were incubated in a blank DMEM medium containing 100 μM of DPPE-PEG-β-CD at 37° C. for 2 hours. After washing off excess DPPE-PEG-β-CD, 150 of μM DPPE-PEG-ADA-modified red blood cells were added and further incubated with resulting hematopoietic stem cells for 2 minutes. After washing off unbound cells, cell-cell conjugates were obtained.

COMPARATIVE EXAMPLE

[0069] Embryonic stem cells were incubated in a blank DMEM medium containing 10 μM of DPPE-PEG-CB[7] at 37° C. for 5 minutes. After washing away excess DPPE-PEG-CB[7], 10 μM of DPPE-PEG-ADA modified doxorubicin-loaded liposomes were added to further incubate for 5 minutes. After washing off the unbound liposomes, fluorescence imaging was performed, and no red fluorescence was found on the embryonic stem cell membrane.

[0070] In summary, the supramolecular cell-based carrier of the embodiment of present disclosure is constructed through host-guest interaction, and is a new generation of cell-based formulations, which can achieve targeted delivery based on the inherent physiological function of transporting cell; The preparation method of supramolecular cell-based carrier is that macrocyclic host molecule coupled with membrane-embedding material anchored on the first cell surface via membrane insertion, and then mixing with the nanoparticles modified with guest molecule or the second cells modified with guest molecule. The preparation process is cell friendly, facile, universal and bioorthogonal, which does not require any covalent modification on the cell surface and has no effect on the physiological function of the modified cell.

[0071] The above descriptions are only preferred examples of the present disclosure and are not intended to limit the present disclosure. For those skilled in the art, there may be various modifications and changes in the present disclosure. Any modifications, equivalent replacements, improvements, etc. made within the spirit and principles of this disclosure shall be included within the protection scope of this disclosure.