Antibody-coupled cell, preparation method and application thereof, and targeted drug delivery system

Abstract

The present invention relates to the technical field of targeted therapeutic drugs, and specifically to an antibody-coupled cell, a preparation method and application thereof, and a targeted drug delivery system. The antibody-coupled cell of the present invention is used in the preparation of a drug carrier. The antibody-coupled cell comprises a cell and an antibody coupled to the cell, and the antibody is used to specifically target antigens on the surface of cells at the disease site. The preparation method of the antibody-coupled cell comprises the steps of: introducing an azide group on the cell surface; n ‑NHS reacts with amino-containing antibodies to obtain alkynyl-containing compounds ‑PEG n ‑mAb, n is an integer from 0 to 100; the alkynyl-containing compound ‑PEG n The mAb is bioorthogonally conjugated to the azide groups on the cell surface to produce antibody-conjugated cells. Antibody-conjugated cells can be used as drug carriers to load drugs. This not only avoids the use of linkers and the need to consider the breakage of drugs and antibodies in target cells, but also greatly simplifies the production process. The reaction is simple and efficient.

Description

Technical Field

[0001] The present invention relates to the technical field of targeted therapeutic drugs, and in particular to an antibody-coupled cell, a preparation method and application thereof, and a targeted drug delivery system. Background Art

[0002] Antibody-Drug Conjugates (ADCs) are a new type of targeted therapeutic drug that achieves precise treatment of diseases by combining highly effective small molecule drugs with specific antibodies.

[0003] ADCs typically consist of three components: a monoclonal antibody (mAb): This is used to target specific cancer cell antigens, ensuring drug specificity and targeting. A linker: This connects the mAb to the small molecule drug (payload), ensuring stability in the bloodstream and effective release upon reaching cancer cells. A small molecule drug: This is a small molecule drug with pharmacological effects that can act on cancer cells.

[0004] ADCs have the following characteristics: Strong targeting: The antibody portion can specifically recognize and bind to antigens on the surface of cells at the disease site, minimizing the impact on normal cells. High efficacy: The small molecule drug portion has highly effective pharmacological effects. Versatility: ADCs combine the targeting of antibodies with the high efficacy of small molecule drugs, resulting in dual therapeutic effects.

[0005] The advantages of ADCs include: Precision therapy: By targeting specific antigens, ADCs can precisely deliver drugs to target cells, minimizing damage to normal cells. Reduced side effects: Due to their specificity and targeting, ADCs have lower systemic side effects than traditional drugs. As an innovative targeted therapy strategy, ADCs combine the targeting of monoclonal antibodies with the high efficacy of small molecule drugs, offering precision, high efficacy, and minimal side effects.

[0006] However, ADC has the following technical defects:

[0007] Linker design: Linkers need to be stable in blood circulation but effectively cleave after reaching the target cells, which makes linker design more difficult.

[0008] Manufacturing cost: The production process of ADC is complex and the cost is high, which limits its widespread application.

[0009] Therefore, methods have been developed to use cell-coupled antibodies as drug carriers. However, these methods are based on thiol modification of cells, which can lead to changes in cell state and thus affect the function of the drug carrier.

[0010] Therefore, the existing technology still needs to be improved and developed. Summary of the Invention

[0011] In view of the above-mentioned deficiencies in the prior art, the present invention aims to provide an antibody-conjugated cell, a preparation method and application thereof, and a targeted drug delivery system, aiming to address the problems of existing ADCs, such as the difficulty in linker design, the complex production process, and the high cost. Furthermore, existing methods for conjugating antibodies to cells require thiol modification of cells, resulting in changes in the cell state and thus affecting the function of the drug carrier.

[0012] The technical solutions of the present invention are as follows:

[0013] In a first aspect, the present invention provides a use of antibody-coupled cells in the preparation of a drug carrier, wherein the antibody-coupled cells comprise cells and antibodies coupled to the cells, wherein the antibodies are used to specifically target antigens on the surface of cells at disease sites;

[0014] The method for preparing the antibody-coupled cells comprises the steps of:

[0015] Extract cells;

[0016] introducing an azide group on the cell surface;

[0017] Alkyne-containing compound-PEG n -NHS reacts with amino-containing antibodies to obtain alkyne-containing compounds -PEG n -mAb, wherein n is an integer from 0 to 100, the alkynyl-containing compound -PEG n -NHS is a polyethylene glycol modified with an alkyne-containing compound and combined with an N-hydroxysuccinimide functional group;

[0018] The alkyne-containing compound-PEG n -mAb is bioorthogonally coupled to the azide group on the cell surface to obtain the antibody-coupled cells.

[0019] Optionally, the cells are isolated from at least one of bone marrow, umbilical cord, umbilical cord blood, placenta, and adipose tissue.

[0020] Optionally, the cells are mesenchymal stem cells, and the cell surface glycoprotein is azidated.

[0021] Optionally, the alkynyl-containing compound is distyryl ring octyl alkynol, the n is 4, the alkynyl-containing compound-PEG n - mAb is DBCO-PEG4-mAb.

[0022] Optionally, the alkynyl-containing compound-PEG n The reaction temperature of -NHS and antibodies containing amino groups is 0~16℃ and the reaction time is 1 minute to 24 hours.

[0023] Further optionally, the alkynyl-containing compound-PEG n The reaction temperature of -NHS and antibodies containing amino groups is 4°C and the reaction time is 1 hour.

[0024] Optionally, the alkyne-containing compound-PEG n The molar ratio of -NHS to the antibody containing amino groups is 100:1 to 1:100.

[0025] Optionally, the alkyne-containing compound-PEG n The temperature for bioorthogonal coupling of the mAb with the azide group on the cell surface is 35-42° C., and the time is 0.5-24 hours.

[0026] Alternatively, the alkynyl-containing compound-PEG n - Bioorthogonal coupling of mAb to the azide groups on the cell surface was performed at 37°C for 1 hour;

[0027] The mass of the alkynyl-containing compound-PEGn-mAb is 0.01-100 nanomoles / 1 million cells.

[0028] The second aspect of the present invention provides a method for preparing antibody-coupled cells, comprising the steps of:

[0029] Extract cells;

[0030] introducing an azide group on the cell surface;

[0031] Alkyne-containing compound-PEG n -NHS reacts with amino-containing antibodies to obtain alkyne-containing compounds -PEG n -mAb, wherein n is an integer from 0 to 100, the alkynyl-containing compound -PEG n -NHS is a polyethylene glycol modified with an alkyne-containing compound and combined with an N-hydroxysuccinimide functional group;

[0032] The alkyne-containing compound-PEG n -mAb is bioorthogonally coupled to the azide groups on the cell surface to obtain antibody-coupled cells.

[0033] The third aspect of the present invention provides an antibody-coupled cell, wherein the antibody-coupled cell is prepared using the method for preparing the antibody-coupled cell of the present invention;

[0034] The antibody-coupled cells include cells and antibodies coupled to the cells, and the antibodies are used to specifically target antigens on the surfaces of cells at disease sites.

[0035] In a fourth aspect, the present invention provides a targeted drug delivery system, comprising a drug carrier and a drug loaded in the drug carrier, wherein the drug carrier is the antibody-coupled cell of the present invention.

[0036] Optionally, the antibody specifically targets antigens on the surface of cells at the disease site, and the drug acts on cells at the disease site.

[0037] Beneficial effects: The present invention provides an antibody-coupled cell, which can be used as a drug carrier to load drugs. This not only avoids the use of linkers and the need to consider the problem of breakage between small molecule drugs and antibodies in target cells, but also greatly simplifies the production process. The antibody only needs to be coupled to the cell using a mild orthogonal reaction. The reaction is simple, efficient, and low-cost; and there is no need to modify the cells with thiol groups. The process is simple and less harmful to the cells, avoiding the cell state changes caused when using thiol-modified cells to construct antibody-coupled cells. In addition, the present invention azidates the cell surface glycoproteins, which can inhibit the function of drug efflux glycoproteins, thereby prolonging the time of the drug in the cell and achieving the effect of drug sustained release. Moreover, when the cells are mesenchymal stem cells, they also have homing properties and can actively move to disease sites such as inflammation. Compared with the passive enrichment of ADCs relying solely on antibodies, the targeting efficiency is higher. BRIEF DESCRIPTION OF THE DRAWINGS

[0038] Figure 1 To convert alkynyl-containing compounds into PEG n -NHS reacts with amino-containing antibodies to produce alkyne-containing compounds -PEG n Schematic diagram of mAb.

[0039] Figure 2 To convert alkynyl-containing compounds into PEG n Schematic diagram of bioorthogonal conjugation of mAb to azide groups on the cell surface to obtain antibody-conjugated cells.

[0040] Figure 3 The figure shows the tissue distribution of CD4 antibody-coupled mesenchymal stem cells in psoriasis mice.

[0041] Figure 4 Figure 4 shows CD4 antibody-coupled mesenchymal stem cells loaded with the RORγt inverse agonist cedirogant (CAS No. 2055496-11-0) for the treatment of psoriasis.

[0042] Figure 5 A graph showing that CD4 antibody-coupled mesenchymal stem cells loaded with the RORγt inverse agonist cedirogant reduce the epidermal thickness of psoriasis mice; where * represents significance, the greater the number of *, the greater the significance and the greater the difference. DETAILED DESCRIPTION

[0043] The present invention provides antibody-coupled cells, their preparation methods and applications, and targeted drug delivery systems. To clarify the objectives, technical solutions, and effects of the present invention, the present invention is described in further detail below. It should be understood that the specific embodiments described herein are intended only to illustrate the present invention and are not intended to limit the present invention.

[0044] An embodiment of the present invention provides an application of antibody-coupled cells in the preparation of a drug carrier, wherein the antibody-coupled cells include cells and antibodies coupled to the cells, wherein the antibodies are used to specifically target antigens on the surface of cells at disease sites;

[0045] The method for preparing the antibody-coupled cells comprises the steps of:

[0046] Extract cells;

[0047] introducing an azide group on the cell surface;

[0048] Alkyne-containing compound-PEG n -NHS reacts with amino-containing antibodies to obtain alkyne-containing compounds -PEG n -mAb, wherein n is an integer from 0 to 100, the alkynyl-containing compound -PEG n -NHS is a polyethylene glycol modified with an alkyne-containing compound and combined with an N-hydroxysuccinimide functional group;

[0049] The alkyne-containing compound-PEG n -mAb is bioorthogonally coupled to the azide group on the cell surface to obtain the antibody-coupled cells.

[0050] In the embodiments of the present invention, autologous cells can be directly extracted from a patient and coupled to antibodies using click chemistry, thereby completing the construction of antibody-coupled cells. Antibody-coupled cells can be used as drug carriers; the desired drug is loaded into the antibody-coupled cells for administration. This targeted drug delivery system can achieve the same targeted drug delivery effect as ADCs, but without the need for a separate ADC design. It can be applied to any existing drug, and has a simple preparation process and low cost. The targeting effect of ADCs is only passive enrichment, while when the cells are mesenchymal stem cells, antibody-coupled mesenchymal stem cells can not only be passively enriched using antibodies but also actively migrate to the site of inflammation by leveraging the active homing effect of mesenchymal stem cells, providing better targeting and achieving high enrichment of antibody-coupled mesenchymal stem cells at the disease site. Furthermore, in the embodiments of the present invention, cell surface glycoproteins are azidated to inhibit the drug's efflux function, thereby extending the drug's time in the cell and achieving a sustained drug release effect. Therefore, the antibody-coupled cells in the embodiments of the present invention not only provide a targeted drug delivery method but also increase the drug's residence time at the target site, potentially improving drug efficacy.

[0051] A detailed description of the method for preparing antibody-coupled cells is provided below and will not be repeated here.

[0052] Among them, antigens on the surface of cells at the disease site refer to substances that are not expressed, expressed at very low or low levels in normal areas, but are expressed at differentially high levels only in the disease site. For example, in mammalian psoriasis, because psoriasis occurs in the skin and its pathological feature is high expression of the surface antigen cluster of differentiation 4 receptor (CD4) in the pathological area, a CD4 antibody-coupled cell can be designed to target diseased skin with high CD4 expression, realizing a simple targeted drug delivery strategy. The antigens on the surface of cells at the disease site can also be antigens on the surface of cancer cells, in which case the antibody specifically targets the antigens on the surface of cancer cells.

[0053] In one embodiment, the cells are isolated from at least one of bone marrow, umbilical cord, cord blood, placenta, adipose tissue, and the like.

[0054] In one embodiment, the cells may be primary cells or cells that have been subcultured for at least one generation, such as mesenchymal stem cells that have been subcultured for 1 to 3 generations. In addition, the mesenchymal stem cells may be mammalian-derived mesenchymal stem cells.

[0055] An embodiment of the present invention provides a method for preparing antibody-coupled cells, comprising the steps of:

[0056] Extract cells;

[0057] introducing an azide group on the cell surface;

[0058] Alkyne-containing compound-PEG n -NHS reacts with amino-containing antibodies to obtain alkyne-containing compounds -PEG n -mAb, where n is an integer from 0 to 100, such as 4, 10, 30, 50, 80, 100, etc. (see Figure 1 );

[0059] The alkyne-containing compound-PEG n -mAb is bioorthogonally coupled to the azide groups on the cell surface to obtain the antibody-coupled cells (see Figure 2 ).

[0060] In one embodiment, the alkyne-containing compound-PEG n -mAb is DBCO-PEG4-mAb.

[0061] DBCO-PEG4-NHS is a polyethylene glycol (PEG) compound modified with a distyryl ring octyl ynol (DBCO) and incorporating an N-hydroxysuccinimide (NHS) functional group. It reacts with an amino-containing antibody (mAb) to produce DBCO-PEG4-mAb. The clickable DBCO-PEG4-mAb is bioorthogonally conjugated to an azide group (-N3) on the surface of mesenchymal stem cells (MSCs), yielding antibody-conjugated MSCs. MSCs are metabolically engineered with N-azidoacetylmannosamine tetraacylation (Ac4ManNAz), which introduces azide groups onto their surfaces.

[0062] In one embodiment, the alkynyl-containing compound-PEG n The temperature for the reaction of -NHS with antibodies containing amino groups is 0~16°C, such as 0°C, 4°C, 6°C, 8°C, 9°C, 10°C, 15°C, 16°C, etc., and the time is 1 minute to 24 hours, such as 1 minute, 10 minutes, 30 minutes, 1 hour, 5 hours, 10 hours, 15 hours, 20 hours, 24 hours, etc.

[0063] In one embodiment, the alkyne-containing compound-PEG n The molar ratio of -NHS to the antibody containing amino groups is 100:1~1:100, such as 1:100, 10:100, 50:100, 1:1, 10:1, 50:1, 80:1, 100:1, etc.

[0064] In one embodiment, the alkyne-containing compound-PEG nThe bioorthogonal conjugation temperature of the mAb to the azide groups on the cell surface is 35-42°C for 0.5-24 hours. Under these conditions, the conjugation of the antibody to the cell is ensured.

[0065] In one embodiment, the alkyne-containing compound-PEG n - The mAb mass is 0.01~100 nanomoles / 1 million cells, such as 0.01 nanomoles, 0.1 nanomoles, 1 nanomoles, 10 nanomoles, 100 nanomoles, etc., to achieve effective coupling between the antibody and cells.

[0066] An embodiment of the present invention provides an antibody-coupled cell, wherein the antibody-coupled cell is prepared using the method for preparing the antibody-coupled cell described in an embodiment of the present invention;

[0067] And / or, the antibody-coupled cells include cells and antibodies coupled to the cells, and the antibodies are used to specifically target antigens on the surface of cells at the disease site.

[0068] An embodiment of the present invention provides a targeted drug delivery system, which includes a drug carrier and a drug loaded in the drug carrier, wherein the drug carrier is the antibody-coupled cell described in the embodiment of the present invention.

[0069] In the embodiments of the present invention, the antibody specifically targets antigens on the surface of cells at the disease site, and the drug acts on cells at the disease site. Antibody-coupled cell-derived small molecule drugs can improve the targeting of small molecule drugs, and antibody-coupled cell-derived drugs can accumulate at the disease site, significantly improving the therapeutic effect.

[0070] In one embodiment, the antibody specifically targets an antigen on the surface of a cancer cell, and the drug acts on the cancer cell.

[0071] The present invention will be further described below with reference to specific examples.

[0072] Example 1

[0073] Taking the mouse psoriasis model as an example, since psoriasis is primarily caused by the skin and its pathological feature is high expression of CD4 at the pathological site, this example designs a CD4 antibody-coupled mesenchymal stem cell that can target diseased skin with high CD4 expression, thereby realizing a simple targeted drug delivery strategy.

[0074] 1. Construction of DBCO-conjugated CD4 antibody

[0075] 1000 molar equivalents of DBCO-PEG4-NHS and 1 molar equivalent of CD4 antibody (CD4mAb) were stirred at 250 rpm in 1 mL of PBS buffer (pH 7.4) at 4°C overnight.

[0076] Because the molecular weight of CD4 antibody is greater than 10Kd, 10KD ultrafiltration tube is used for multiple low-temperature centrifugation to remove the unreacted DBCO-PEG4-NHS to obtain pure DBCO-coupled CD4 antibody (DBCO-PEG4-CD4mAb).

[0077] Construction of antibody-coupled mesenchymal stem cells

[0078] Specific pathogen-free (SPF) C57BL / 6J mice aged 3-4 weeks were sacrificed by cervical dislocation and then immersed in 75% ethanol.

[0079] The outer and inner skin of the mouse's legs were then cut open, and the femurs and tibiae were removed under sterile conditions and immersed in sterile PBS buffer containing double-antibody antibodies. The surrounding muscle and connective tissue of the femurs and tibiae were removed and washed three times with PBS buffer containing double-antibody antibodies.

[0080] Use ophthalmic scissors to cut off the ends of the femur and tibia to expose the medullary cavity.

[0081] Collect the femur and tibia after flushing the bone marrow into a sterile culture dish, and carefully cut the bone tissue into about 1-3 mm with sterile ophthalmic scissors. 3 fragments.

[0082] Add an appropriate amount of fresh complete culture medium (containing 5% fetal bovine serum, FBS) containing 1 mg / mL type II collagenase, and shake in a shaker at 37°C for digestion.

[0083] After digestion, centrifuge and discard the supernatant. Wash twice with PBS buffer, then inoculate bone fragments into culture dishes, add appropriate amount of fresh complete culture medium (containing 20% ​​FBS) to submerge the bone fragments, and culture in a 37°C 5% carbon dioxide incubator.

[0084] After 72 hours of culture, perform the first half-volume medium change. Change the medium every 3 days thereafter. When adherent cells reach 80%-90% confluence, proceed with cell expansion and passage.

[0085] 2*10 4 Mouse bone marrow mesenchymal stem cells (BMSCs) were seeded into 35 mm cell culture dishes and cultured for 24 hours. Then, 10 μM Ac4ManNAz was added and co-cultured with the cells for 24 hours.

[0086] The original culture medium was discarded, and the cells were washed three times with PBS buffer. Then, 100 nM DBCO-PEG4-CD4 mAb was added and reacted at 37°C for 1 hour to obtain CD4 antibody-coupled mesenchymal stem cells (also referred to as CD4mAb-BMSCs).

[0087] 3. Distribution of CD4 antibody-coupled mesenchymal stem cells in psoriasis mice

[0088] Healthy male Balb / c mice aged 7-8 weeks were selected and their backs were shaved with an area of ​​2.5 cm×2.5 cm.

[0089] Imiquimod (IMQ) ointment (purchased from Sichuan Mingxin Pharmaceutical Co., Ltd.) was applied to the back skin of mice for 5 consecutive days to establish a psoriasis mouse model. The control group was given the same dose of vaseline ointment.

[0090] On the second day of modeling, CD4 antibody-coupled BMSCs and untreated BMSCs were stained with cell membrane far-infrared probe (DiD) dye for cell membrane fluorescence. Then, blank control mesenchymal stem cells and CD4 antibody-coupled mesenchymal stem cells (cells were placed in 200 μL of normal saline, and the number of cells was 2*10 5 ), the DiD fluorescence distribution of mesenchymal stem cells in mice was investigated using in vivo imaging (IVIS) equipment at 24, 72, and 96 hours.

[0091] Figure 3 It was shown that 24 hours after administration, neither the control mesenchymal stem cells (BMSCs) nor the CD4 antibody-coupled mesenchymal stem cells (CD4mAb-BMSCs) were distributed in the skin. However, 72 hours after administration, the enrichment of CD4 antibody-coupled mesenchymal stem cells in the diseased areas of the skin increased significantly compared with the control group. After 96 hours, the control mesenchymal stem cells could no longer be observed in the skin, while the CD4 antibody-coupled mesenchymal stem cells were still highly enriched in the diseased areas of the skin.

[0092] Therefore, the antibody-coupled mesenchymal stem cells of the present invention can not only provide a targeted drug delivery method, but also increase the residence time of the drug at the target site, thereby having the potential to improve drug efficacy.

[0093] 4. CD4 antibody-coupled mesenchymal stem cells carrying RORγt inverse agonists for the treatment of psoriasis

[0094] Healthy male Balb / c mice aged 7-8 weeks were selected and their backs were shaved with an area of ​​2.5 cm×2.5 cm.

[0095] Imiquimod (IMQ) ointment (Sichuan Mingxin Pharmaceutical Co., Ltd.) was applied to the back skin of mice for 7 consecutive days to establish a psoriasis mouse model.

[0096] On the second day of modeling, small molecule drugs cedirogant (abbreviated as cdg) as a control, mesenchymal stem cells (BMSCs), CD4 antibody-coupled mesenchymal stem cells (CD4mAb-BMSCs), and CD4 antibody-coupled mesenchymal stem cells carrying RORγt inverse agonist cedirogant (cdg@CD4mAb-BMSCs) were injected through the tail vein (all drugs were placed in 200uL normal saline, and the number of BMSCs was 5*10 5 ), and the drug's treatment effect on psoriasis skin was investigated on the seventh day.

[0097] Figure 4 It was shown that the skin damage, rash and desquamation of mice in the model group increased. Figure 5 The results showed that the epidermis of the model group was thickened. Figure 4-Figure 5 The study showed that the small molecule drug cdg can partially improve skin damage in mice. CD4 antibody-coupled mesenchymal stem cells loaded with cdg can accumulate at the site of skin damage, significantly improving skin damage in mice and inhibiting epidermal thickening. The therapeutic effect is significantly better than the group injected with cdg alone, indicating that antibody-coupled mesenchymal stem cells loaded with small molecule drugs can improve the targeting of small molecule drugs, thereby improving their efficacy.

[0098] In summary, the present invention provides an antibody-coupled mesenchymal stem cell, its preparation method and application, and targeted drug delivery system. The present invention can directly extract autologous mesenchymal stem cells from the patient, and use click chemistry to couple antibodies to complete the construction of antibody-coupled mesenchymal stem cells. Antibody-coupled mesenchymal stem cells can be used as drug carriers. When administering, the required drug is loaded into the antibody-coupled mesenchymal stem cells for administration. This targeted drug delivery system can achieve the same targeted drug delivery effect as ADC, but there is no need to design an ADC separately. Instead, it can be applied to any existing drug, and the preparation process is simple and the price is low. The targeting effect of ADC is only passive enrichment, while antibody-coupled mesenchymal stem cells can not only be passively enriched using antibodies, but can also actively move to the site of inflammation using the active homing effect of mesenchymal stem cells, with better targeting. In addition, the present invention azidoates cell surface glycoproteins, which can inhibit the function of drug excretion glycoproteins, thereby prolonging the time of the drug in the cell, and then achieving the effect of drug sustained release.

[0099] It should be understood that the application of the present invention is not limited to the above examples. For those skilled in the art, improvements or changes can be made based on the above description. All these improvements and changes should fall within the scope of protection of the claims attached to the present invention.

Claims

1. A targeted drug delivery system, characterized in that: The invention comprises a drug carrier and a small molecule drug loaded in the drug carrier, wherein the drug carrier is an antibody-coupled cell, and the antibody-coupled cell comprises a cell and an antibody coupled to the cell, wherein the antibody specifically targets antigens on the surface of cells at the disease site, so that the small molecule drug acts on the cells at the disease site; The method for preparing the antibody-coupled cells comprises the steps of: Extract cells; introducing an azide group on the cell surface; Alkyne-containing compound-PEG n -NHS reacts with amino-containing antibodies to obtain alkyne-containing compounds -PEG n -mAb, wherein n is 4, the alkynyl-containing compound -PEG n -NHS is a polyethylene glycol modified with an alkyne-containing compound and combined with an N-hydroxysuccinimide functional group; The alkyne-containing compound-PEG n - performing bioorthogonal coupling of the mAb to the azide groups on the cell surface to obtain the antibody-coupled cells; The cells are mesenchymal stem cells, the cell surface glycoproteins are azidated, and the cells are isolated from at least one of bone marrow, umbilical cord, umbilical cord blood, placenta, and adipose tissue; The small molecule drug is cedirogant, and the disease site is a psoriasis skin disease site.

2. The targeted drug delivery system according to claim 1, characterized in that The alkynyl group-containing compound is distyryl cyclooctyl ynol.

3. The targeted drug delivery system according to claim 1, wherein: The alkyne-containing compound-PEG n -NHS reacts with amino-containing antibodies at a temperature of 0-16°C for a period of 1 minute to 24 hours; The alkyne-containing compound-PEG n The molar ratio of -NHS to the antibody containing an amino group is 1:100 to 100:

1.

4. The targeted drug delivery system according to claim 1, wherein: The alkyne-containing compound-PEG n - The bioorthogonal coupling of mAb to the azide groups on the cell surface is carried out at a temperature of 35 to 42°C for 0.5 to 24 hours; The alkyne-containing compound-PEG n -mAb mass ranges from 0.01 to 100 nanomoles per million cells.

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